Treatment of sepsis and septic shock

Abstract

The present invention relates to a composition comprising a mixture of empty liposomes wherein the mixture of empty liposomes comprises: (a) a first empty liposome comprising cholesterol wherein the amount of cholesterol is at least 30% (weight / weight); and (b) a second empty liposome comprising sphingomyelin, for use in the treatment of sepsis, severe sepsis, septic shock or long-term and severe hypotension, preferably persistent hypotension, in an animal, preferably a human, for use in the treatment of hyperlipidemia in an animal, preferably a human. The present invention relates to a pharmaceutical composition for treating toxic shock syndrome in animals, preferably humans, or to the treatment of toxic shock syndrome in animals, preferably humans, or to the treatment of hypotension in acute lung injury, preferably said persistent hypotension, or to the treatment of toxic shock syndrome in animals, preferably humans, preferably septicaemia, severe septicaemia, acute respiratory distress syndrome, or to the treatment of hypotension in acute lung injury, preferably said persistent hypotension.

Description

[0001] This application is a divisional application of patent application No. 201980025257.3, filed on April 18, 2019, entitled “Treatment of sepsis and septic shock”.

[0002] This invention relates to a composition comprising a mixture of empty liposomes, wherein the mixture of empty liposomes comprises: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin, the composition being used to treat sepsis, preferably in animals or humans, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, in animals or humans, preferably in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or hypotension in acute lung injury, preferably persistent hypotension, or to treat animals or animals, preferably in animals, preferably in humans, toxic shock syndrome. Specifically, this invention relates to compositions for treating sepsis or septic shock in animals, preferably in humans. Background Technology

[0003] Sepsis is a potentially life-threatening organ dysfunction caused by a dysregulated host response to infection. It is characterized by physiological, pathological, and biochemical abnormalities leading to organ dysfunction (Levy M et al., *Intensive Care Medicine*, 2003; 29:530-38; Singer M et al., *JAMA*, 2016; 315(8):801-810). Sepsis can lead to tissue damage, multi-system organ failure, and subsequently death (Cohen, 2002, *Nature*, 420, 885-891). Despite maximal care, approximately 30% to 50% of patients with sepsis die. The underlying mechanisms of this systemic inflammatory dysregulation are complex and may involve multiple pathways. Therefore, antimicrobial agents alone cannot effectively treat sepsis. In fact, although antibiotics are available, current treatments for sepsis have proven ineffective. Neutralization of specific inflammatory cytokines was also ineffective, highlighting the need for new therapies (Wenzel and Edmond, 2012, New England Journal of Medicine 366, 2122-2124).

[0004] Septic shock occurs in a subset of patients with sepsis and is associated with increased mortality. The pathophysiology of septic shock is not fully understood; it includes underlying circulatory and cellular / metabolic abnormalities severe enough to significantly increase mortality. Patients with septic shock can be identified by the clinical constructs of sepsis characterized by: persistent hypotension requiring vasopressors to maintain a mean arterial pressure of 65 mm Hg or higher despite adequate volume resuscitation; organ inadequacy; and serum lactate levels above 2 mmol / L (18 mg / dL) (Singer M et al., JAMA 2016; 315(8):801-10).

[0005] Sepsis and septic shock have lasting effects on patients. For example, prolonged tissue hypoperfusion can lead to long-term neurological and cognitive sequelae.

[0006] In addition to bacterial infections, viral infections also increase susceptibility to infections that can lead to sepsis or septic shock by enhancing susceptibility to bacterial co-infections. For example, influenza patients frequently exhibit increased susceptibility to Streptococcus pneumoniae co-infections, and sepsis has been reported as a leading cause of death during influenza pandemics. The detailed mechanisms by which viral infections susceptible patients to bacterial infections and subsequently lead to sepsis are not fully understood, and therapies for treating and preventing sepsis and septic shock in patients with viral infections are still needed.

[0007] In recent years, customized empty liposomes, such as those composed of cholesterol and / or sphingomyelin, and their use in the treatment of bacterial infections have been described as trapping virulence factors such as bacterial toxins, enzymes, and toxic appendages (WO 2013 / 186286; Henry BD et al., Nature Biotechnol 2015; 33(1):81-88; Azeredo da Silveira, S and Perez, A, Expert Rev. AntiInfect Ther. 2015; 13(5):531-533; Azeredo da Silveira, S and Perez, A. Expert Rev. AntiInfect Ther. 2017; 15:973-975). These custom-designed empty liposomes have also been described as exhibiting antiviral activity and thus as a therapy for antiviral infections, particularly as a therapy for neutralizing enveloped viruses such as influenza virus (WO 2017 / 216282). Summary of the Invention

[0008] In the first human studies in patients with severe pneumonia, the preferred compositions of the present invention showed surprisingly positive results in improving clinical signs and symptoms. Furthermore, and importantly, it was surprisingly found that the preferred compositions of the present invention improved hemodynamic parameters, prevented hemodynamic deterioration, and accelerated the resolution of septic shock. Therefore, as revealed by accelerating the normalization of hemodynamic instability, thereby leading to patient recovery and faster discharge from the intensive care unit (ICU), the compositions of the present invention are effective in treating sepsis and septic shock. Based on this promising efficacy data and its credible mechanism of action, further efficacy studies in patients suspected or confirmed to have an infection, regardless of the pathogen, and exhibiting signs of complications or progression of severity, particularly community-acquired pneumonia, hospital-acquired pneumonia, ventilation-associated pneumonia, intra-abdominal infections, skin and soft tissue infections, urinary tract infections, or bacteremia, are reasonable. Furthermore, based on these promising efficacy data and its credible mechanism of action, the compositions of the present invention are particularly considered to be highly beneficial for the treatment and prevention of sepsis or septic shock caused or associated with bacterial or viral pathogens that use specific lipid microstructural domains to attack the host, and thereby attack animals, preferably human patients.

[0009] Therefore, in a first aspect, the present invention provides a composition comprising a mixture of empty liposomes, preferably consisting of the mixture of said empty liposomes, wherein said mixture of empty liposomes comprises, preferably consists of: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; said composition is used to treat sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension in animals, preferably humans, septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or hypotension in acute lung injury, preferably persistent hypotension, or to treat toxic shock syndrome in animals, preferably humans.

[0010] In another aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is intended for treating sepsis, preferably in humans, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension.

[0011] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is intended for treating septic shock in animals, preferably humans.

[0012] As this description continues, other aspects and embodiments of the invention will become apparent. Attached Figure Description

[0013] Figure 1: Evolution of cardiovascular SOFA scores from baseline (before administration) to day 8 in the entire study population, presented as absolute values, across the placebo (diamond), low-dose CAL02 (triangle), and high-dose CAL02 (square) groups. Figure 1A The evolution of the cardiovascular SOFA score from baseline (before administration) to day 8 in the entire study population, presented as the difference from baseline score, excluding patients without any hypotensive events (three patients in the CAL02 high-dose group). Figure 1 B *p<0.05

[0014] Figure 2 The evolution of the cardiovascular SOFA score from baseline (before administration) to day 8 in patients already in septic shock, presented as absolute values ​​in the placebo (diamond), low-dose CAL02 (triangle), and high-dose CAL02 (square) groups. Detailed Implementation

[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 pertains.

[0016] As used herein, the term "about" should have a meaning of + / - 5%. For example, about 50% should mean 47.5% to 52.5%. Preferably, the term "about" as used herein should have a meaning of + / - 3%. For example, about 50% should mean 48.5% to 51.5%.

[0017] Unless otherwise stated, when the term "an" or "a" is used herein, it means "at least one / a type". Specifically, the term "an" or "a" used in conjunction with the single empty liposome, the first empty liposome, and the second empty liposome to describe the empty liposomes and mixtures of empty liposomes according to the invention should typically and preferably refer to a single empty liposome and a mixture of empty liposomes including the first empty liposome and the second empty liposome.

[0018] The range of values ​​disclosed herein should refer to all values ​​within the range, including the value that defines the range. For example, by illustration, a value of 12 to 13 should refer to the value 12 or 13, or all values ​​between 12 and 13.

[0019] As used herein, the term "empty liposome" refers to a liposome having an average diameter of 20 nm to 10 μm, preferably 20 nm to 500 nm, and more preferably 20 nm to 400 nm, and even more preferably 40 nm to 400 nm or 20 nm to 200 nm, preferably an artificial liposome, and composed of one or more phospholipid bilayers, and typically and preferably monolayer vesicles and multilayer vesicles, more preferably small monolayer vesicles (SUVs). In preferred embodiments, the term "empty liposome" as used herein typically and preferably refers to a liposome without any drug inclusion, typically and preferably refers to a liposome without any pharmaceutical drug inclusion. As used herein and when referring to the empty liposomes of the present invention, "incorporated / incorporating" typically and preferably means encapsulated / encapsulating within the cavity of the liposome, within a potential bilayer of the liposome, or as part of the membrane layer of the liposome. In another preferred embodiment, as used herein, the term "empty liposome" typically and preferably refers to liposomes according to the invention composed of sphingomyelin and cholesterol or composed of sphingomyelin, and further comprises only water-soluble inorganic compounds and / or water-soluble organic molecules, wherein typically and preferably, the water-soluble inorganic compounds and / or water-soluble organic molecules are derived from the synthesis of the empty liposomes of the invention, and wherein typically and preferably, the water-soluble inorganic compounds are inorganic salts, preferably selected from NaCl, KCl, and MgCl2, and wherein the water-soluble organic molecules are buffers, wherein preferably, the water-soluble organic molecules are selected from glucose and HEPES. Typically and preferably, the water-soluble inorganic compounds and / or water-soluble organic molecules are incorporated into the empty liposomes of the invention due to their presence during the production of the empty liposomes of the invention. In another preferred embodiment, as used herein, the term "empty liposome" typically and preferably refers to liposomes according to the invention composed of sphingomyelin and cholesterol or composed of sphingomyelin, and wherein the empty liposomes do not include antioxidants.In a further preferred embodiment, as used herein, the term "empty liposome" typically and preferably refers to liposomes according to the invention composed of sphingomyelin and cholesterol or composed of sphingomyelin, and further comprises only water-soluble inorganic compounds and / or water-soluble organic molecules, wherein typically and preferably, the water-soluble inorganic compounds and / or water-soluble organic molecules are derived from the synthesis of the empty liposomes of the invention, and wherein typically and preferably, the water-soluble inorganic compounds are inorganic salts, preferably selected from NaCl, KCl and MgCl2, and wherein the water-soluble organic molecules are buffers, wherein preferably, the water-soluble organic molecules are selected from glucose and HEPES, and wherein the empty liposomes according to the invention composed of sphingomyelin and cholesterol or composed of sphingomyelin do not contain antioxidants.

[0020] Sepsis or septic shock: Sepsis is a disease with an infectious cause and a pathology of systemic inflammatory response syndrome (SIRS); it is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection (Levy M et al., Intensive Care Medicine, 2003; 29:530-38; Singer M et al., JAMA, 2016; 315(8):801-810). Initial symptoms include chills, sweating, fever, and hypotension. Microcirculatory disturbances occur as various inflammatory mediators and clotting factors increase throughout the body, leading to a worsening of the pathological condition. Septic shock involves abnormal organ perfusion, uncontrolled hypotension, and multiple organ dysfunction, which can lead to death. The clinical profile of sepsis is characterized by: persistent hypotension requiring vasopressors to maintain a mean arterial pressure (MAP) ≥ 65 mm Hg, despite adequate volume resuscitation; and serum lactate levels > 2 mmol / L (18 mg / dL). Therefore, the term “sepsis” as used herein should refer to life-threatening organ dysfunction resulting from a dysregulated host response to infection, as defined and recommended in Singer M et al., JAMA. 2016; 315(8):801-810 (Recommendation; Column 3). Organ dysfunction can be identified as a sharp change in the total SOFA (Sequential [Sepsis-Related] Organ Failure Assessment) score ≥2 after infection. For patients with no known pre-existing organ dysfunction, the baseline SOFA score can be assumed to be zero. A SOFA score ≥2 reflects an overall mortality risk of approximately 10% in a general hospital population with suspected infection. Even patients presenting with moderate dysfunction may deteriorate further, highlighting the severity of the condition and the need for timely and appropriate intervention (if such intervention has not yet been implemented). If a suspected infected patient with a high risk of prolonged ICU stay or death in the hospital meets at least two of the following clinical criteria that collectively constitute a new bedside clinical score (called the rapid SOFA (qSOFA)), the patient can be rapidly identified as more likely to have a sepsis-typical adverse outcome: a respiratory rate of 22 / min or higher; altered mental activity, i.e., altered mental status; or a systolic blood pressure of 100 mm Hg or lower. As used herein and as defined and recommended by Singer M et al., JAMA. 2016; 315(8):801-810 (recommendation; Column 3), the term “septic shock” is a subtype of sepsis in which underlying circulatory and cellular / metabolic abnormalities are severe enough to significantly increase mortality, and therefore, the risk of death associated with septic shock is higher than that of sepsis alone.Patients with septic shock can be identified by the clinical constructs of sepsis characterized by: persistent hypotension requiring vasopressors to maintain a mean arterial pressure (MAP) ≥65 mm Hg, despite adequate volume resuscitation (and therefore in the absence of hypovolemia); and serum lactate levels >2 mmol / L (18 mg / dL). Under these criteria, hospital mortality exceeds 40%.

[0021] Animals: As used herein, the term “animal” refers to a living multicellular vertebrate organism, including, for example, mammals and birds. The term mammal includes both human and non-human mammals. Similarly, the term “subject” includes both human and livestock subjects.

[0022] As used herein, the terms “treating” or “therapy” refer to means of achieving a desired physiological effect. An effect can be therapeutic in terms of partially or completely curing a disease or symptom and / or symptoms attributable to said disease or symptom, including improving signs or symptoms of a disease such as sepsis or septic shock, or pathological conditions associated with said disease, such as reducing fever in a subject with septic shock or stabilizing blood pressure in said subject, or improving symptoms such as, but not limited to, chills, sweating, or increasing organ function. As used herein in its broadest sense, the term “treatment” should include and refer to the “prevention” of a disease. “Preventing” or “prevention” of a disease refers to suppressing a disease or symptom such as sepsis or septic shock, i.e., preventing the partial or complete development of a disease such as sepsis or septic shock in a person, for example, suffering from or at risk of bacterial infection. Therefore, the term “treatment” as used herein, for the purpose of defining and characterizing preferred aspects and embodiments of the invention, should be excluded from and should not refer to the “prevention” of a disease in its preferred meaning. Furthermore, in other embodiments and aspects of the invention’s prevention, particularly in animals, and preferably in humans at risk of sepsis or septic shock, the compositions used and the methods of the invention are subsequently used to delay or prevent the development of sepsis or septic shock. Thus, the method comprises: selecting a human patient at risk of sepsis or septic shock typically and preferably caused by infection; and administering one or more of the compositions disclosed herein to said human patient. The human patient may be, for example, an intubated person, i.e., a person under invasive mechanical ventilation, or a person who has been exposed to specific bacteria such as Streptococcus pneumoniae or Staphylococcus aureus.

[0023] Singer et al., in JAMA 2016, well described the predictive power of patients potentially suffering from sepsis: any two of the three clinical variables—a Glasgow Coma Scale score of 13 or lower; a systolic blood pressure of 100 mmHg or lower; and a respiratory rate of 22 / min or higher—provided similar predictive power as the full SOFA score outside the ICU. Based on external US and non-US datasets, this model demonstrated robustness in multiple sensitivity analyses, including simpler assessments of altered mental status (Glasgow Coma Scale score <15), performed in out-of-hospital, emergency room, and ward settings. For patients suspected of infection within the ICU, the SOFA score was superior to this model, potentially reflecting the moderating effect of interventions (e.g., vasopressors, sedatives, mechanical ventilation).

[0024] "Therapeutic effective dose" is the amount of composition that achieves the desired effect in the treated subject. For example, this could be the amount necessary to suppress septic shock, reduce fever, or prevent multiple organ failure in animals, preferably human patients, such as patients with pneumonia and / or patients infected with Streptococcus pneumoniae. When administered to animals, preferably human patients, a dose that will achieve an effective target tissue concentration will typically be used.

[0025] As used herein, “therapeutic dose” refers to a dose known to those skilled in the art to have a therapeutic effect.

[0026] As used in this article, the term “pneumonia” should encompass “community-acquired pneumonia” (CAP), “hospital-acquired pneumonia” (HAP), or “ventilator-associated pneumonia” (VAP).

[0027] The terms “community-acquired pneumonia” or “CAP” are known to those skilled in the art, see, for example, the IDSA / ATS Guidelines for CAP in Adults (CID 2007:44(Supplement 2)S27). Specifically, the term refers to pneumonia acquired outside of a hospital.

[0028] The term “hospital-acquired pneumonia (HAP)” refers to pneumonia acquired during or after hospitalization for another illness or surgery, which occurs within at least 48 to 72 hours of admission.

[0029] As defined herein, “ventilator-associated pneumonia (VAP)” is pneumonia that develops 48 hours or longer after mechanical ventilation and is characterized by microbial invasion of the lower respiratory tract and lung parenchyma. VAP is a potentially serious medical condition.

[0030] Pneumonia is caused by a variety of microbial infections, including bacteria such as Streptococcus pneumoniae, Haemophilus influenzae, Legionella pneumophila, Staphylococcus aureus, and Pseudomonas aeruginosa (for CAP), and Gram-negative bacilli such as Pseudomonas aeruginosa and Serratia marcescens (for HAP), as well as Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Stenotrophomonas maltophilia, Acinetobacter species, and Haemophilus influenzae (see Cilloniz et al., Thorax, April 2011; 66(4):340-6 and Jones RN. Clinical Infectious Diseases). Dis) August 2010; 51 (Supplement 1): S81-7).

[0031] The term “severe community-acquired pneumonia” or “sCAP” is known to those skilled in the art. Specifically, “sCAP” refers to a subgroup of patients with community-acquired pneumonia who require intensive care. The Infectious Diseases Society of America (IDSA) and the American Thoracic Society (ATS) have published guidelines on 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(Supplement 2), Table 4). According to the IDSA / ATS guidelines, sCAP is defined as CAP requiring intensive care. Admission to the intensive care unit is recommended if a CAP patient exhibits one or both of the two major criteria described in Example 1 and implemented in the presented studies, or if three minor criteria from the stated list are present.

[0032] As used herein, the term "for" in "compositions for treating diseases" should also disclose the corresponding therapeutic method and the corresponding use in preparing a formulation for treating diseases.

[0033] In one aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition being used to treat septic shock in animals, preferably humans.

[0034] In another aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is intended for treating hypotension, preferably persistent hypotension, in animals, preferably humans, wherein the hypotension, preferably persistent hypotension, is associated with septic shock.

[0035] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is used to treat sepsis, preferably in humans, sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, in animals, preferably in humans, septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or hypotension in acute lung injury, preferably persistent hypotension, or to treat toxic shock syndrome in animals, preferably in humans.

[0036] In another aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is intended for treating sepsis, preferably in humans, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension.

[0037] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (by weight); and (b) a second empty liposome comprising sphingomyelin; the composition being used to treat sepsis or severe sepsis in animals, preferably humans.

[0038] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is intended for treating septic shock in animals, preferably humans.

[0039] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (by weight); and (b) a second empty liposome comprising sphingomyelin; the composition being used to treat long-term and severe hypotension, preferably persistent hypotension, in animals, preferably humans.

[0040] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is intended for treating hypotension in animals, preferably humans, with septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or acute lung injury, preferably persistent hypotension.

[0041] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is used to treat persistent hypotension in septic shock in animals, preferably humans.

[0042] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is used to treat hypotension in animals, preferably humans, with septic shock, preferably with persistent hypotension.

[0043] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is used to treat persistent hypotension in septic shock in humans.

[0044] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition being used to treat hypotension in sepsis or severe sepsis in animals, preferably humans, preferably persistent hypotension.

[0045] In a further aspect, the present invention provides a composition comprising, preferably, a mixture of empty liposomes, wherein the mixture of empty liposomes comprises, preferably, the following: (a) a first empty liposome comprising cholesterol, wherein the amount of cholesterol is at least 30% (w / w); and (b) a second empty liposome comprising sphingomyelin; the composition is intended for treating toxic shock syndrome in animals, preferably humans.

[0046] In a further aspect and in a very preferred embodiment, the present invention provides a composition for treating hypotension, preferably persistent hypotension, in the animal, preferably the human, suffering from sepsis. In a further aspect and in a very preferred embodiment, the present invention provides a composition for treating hypotension, preferably persistent hypotension, in the human suffering from sepsis. In a further aspect and in a very preferred embodiment, the present invention provides a composition for treating persistent hypotension in the human suffering from sepsis.

[0047] In a further aspect and in a very preferred embodiment, the present invention provides a composition for treating hypotension, preferably persistent hypotension, in the animal, preferably the human, suffering from septic shock. In a further aspect and in a very preferred embodiment, the present invention provides a composition for treating hypotension, preferably persistent hypotension, in the human suffering from septic shock. In a further aspect and in a very preferred embodiment, the present invention provides a composition for treating persistent hypotension in the human suffering from septic shock.

[0048] In another aspect, the present invention provides a method for treating sepsis or septic shock in animals, preferably humans, in need of sepsis, preferably septic shock, the method comprising administering a therapeutically effective amount of the composition as defined in the appended claims. Preferably, the human patient suffers from pneumococcal pneumonia, or the sepsis or septic shock is preferably caused by pneumococcal pneumonia.

[0049] For all aspects and embodiments disclosed herein, therapeutically effective amounts of the compositions of the invention are typically and preferably used for the disclosed treatments.

[0050] Furthermore, all embodiments and preferred embodiments disclosed herein should be understood as embodiments and preferred embodiments of any and all aspects of the present invention.

[0051] The first study compared the preferred composition of the present invention plus standard antibiotic therapy with placebo plus 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 showed a synergistic effect between the preferred composition of the present invention (designated CAL02) and antibiotic treatment.

[0052] As a result, the preferred compositions of the present invention thus capture and neutralize toxins released from a variety of bacteria associated with serious infections in a synergistic manner with antibiotic treatment in human patients. The preferred compositions of the present invention are effective regardless of the resistance profile of the target pathogen and do not induce resistance.

[0053] In a preferred embodiment, the second empty liposome (b) comprises sphingomyelin as the sole lipid component. In a preferred embodiment, the second empty liposome (b) is composed of sphingomyelin.

[0054] In a preferred embodiment, the amount of cholesterol in the first empty liposome (a) is 30% to 70% (w / w), and preferably the amount of cholesterol in the first empty liposome (a) is 35% to 60% (w / w).

[0055] In a preferred embodiment, the amount of cholesterol in the empty liposome (a) is 45% to 55% (by weight), and preferably the amount of cholesterol in the empty liposome (a) is about 50% (by weight).

[0056] In a preferred embodiment, the first empty liposome (a) is composed of cholesterol and sphingomyelin, and wherein preferably the amount of cholesterol in the empty liposome (a) is 45% to 55% (w / w), and further preferably the amount of cholesterol in the empty liposome (a) is about 50% (w / w).

[0057] In a preferred embodiment, the first empty liposome (a) comprises the cholesterol and the sphingomyelin as the sole lipid components.

[0058] In a preferred embodiment, the amount of cholesterol in the empty liposome (a) is 45% to 55% (by weight), and preferably the amount of cholesterol in the empty liposome (a) is about 50% (by weight), and wherein the second empty liposome (b) is composed of sphingomyelin.

[0059] In a preferred embodiment, the first empty liposome (a) is composed of cholesterol and sphingomyelin, and the amount of cholesterol in the empty liposome (a) is 45% to 55% (w / w), and preferably the amount of cholesterol in the empty liposome (a) is about 50% (w / w), and the second empty liposome (b) is composed of sphingomyelin.

[0060] In a preferred embodiment, the first empty liposome (a) is composed of cholesterol and sphingomyelin, and the amount of cholesterol in the empty liposome (a) is 45% to 55% (w / w), and preferably the amount of cholesterol in the empty liposome (a) is about 50% (w / w), and the first empty liposome (a) includes the cholesterol and the sphingomyelin as the only lipid components, and the second empty liposome (b) is composed of sphingomyelin.

[0061] In a preferred embodiment, the mixture of empty liposomes comprises at least 20% (by weight) of the first empty liposome (a) and the second empty liposome (b), and wherein preferably the mixture of empty liposomes comprises at least 30% (by weight) of the first empty liposome (a) and the second empty liposome (b).

[0062] In a preferred embodiment, the mixture of empty liposomes comprises at least 40% (by weight) of the first empty liposome (a) and the second empty liposome (b).

[0063] In a preferred embodiment, the first empty liposome (a) is composed of cholesterol and sphingomyelin, and further preferably the amount of cholesterol in the empty liposome (a) is about 50% (by weight), and the mixture of the empty liposomes comprises at least 40%, preferably at least 45% (by weight) of the first empty liposome (a) and the second empty liposome (b).

[0064] In a preferred embodiment, the first empty liposome (a) is composed of a 1:1 (weight / weight-w / w) mixture of the first empty liposome and the second liposome, wherein the first empty liposome is composed of cholesterol and sphingomyelin in a 1:1 weight ratio (1:1 w / w; molar ratio 35:65), and the second empty liposome is composed of sphingomyelin alone.

[0065] In a preferred embodiment, the first empty liposome (a) is composed of a 1:1 (weight / weight-w / w) mixture of the first empty liposome and the second liposome, wherein the first empty liposome is composed of cholesterol and sphingomyelin in a 1:1 weight ratio (1:1 w / w; molar ratio 35:65), and the second empty liposome is composed of sphingomyelin alone, wherein the first empty liposome (a) includes the cholesterol and the sphingomyelin as the sole lipid components, and the second empty liposome (b) includes the sphingomyelin as the sole lipid component.

[0066] In a preferred embodiment, the first empty liposome has an average diameter of about 130 nm, and the second empty liposome has an average diameter of about 90 nm.

[0067] In a preferred embodiment, the composition is used to treat hypotension in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome or acute lung injury, preferably persistent hypotension, preferably for treating hypotension in septic shock, preferably persistent hypotension, wherein the hypotension, preferably persistent hypotension, is associated with systolic blood pressure <90 mmHg or mean arterial pressure <70 mmHg.

[0068] In a preferred embodiment, the composition is used to treat hypotension in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome or acute lung injury, preferably persistent hypotension, preferably for treating hypotension in septic shock, preferably persistent hypotension, wherein the hypotension, preferably the persistent hypotension, is associated with a systolic blood pressure <90 mmHg.

[0069] In a preferred embodiment, the composition is used to treat hypotension, preferably persistent hypotension, in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome or acute lung injury, and is preferably used to treat hypotension in septic shock, preferably persistent hypotension, wherein the hypotension, preferably persistent hypotension, is associated with a mean arterial pressure <70 mmHg.

[0070] In a preferred embodiment, the composition is used to treat hypotension, preferably persistent hypotension, in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or acute lung injury. The hypotension is associated with a systolic blood pressure <90 mmHg or a mean arterial pressure <70 mmHg, and the hypotension is pretreated with a vasopressor for at least 2 hours.

[0071] In a preferred embodiment, the composition is used to treat hypotension, preferably persistent hypotension, in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or acute lung injury. The hypotension is associated with a systolic blood pressure <90 mmHg or a mean arterial pressure <70 mmHg, and the hypotension, preferably persistent hypotension, is pretreated with vasopressors for at least 2 hours after fluid resuscitation.

[0072] In a preferred embodiment, the composition is used to treat hypotension in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or acute lung injury, preferably persistent hypotension, preferably for treating hypotension in septic shock, preferably the persistent hypotension, wherein the hypotension, preferably the persistent hypotension, is associated with systolic blood pressure <90 mmHg or mean arterial pressure <70 mmHg, and wherein the hypotension, preferably the persistent hypotension, is pretreated with at least one, preferably a therapeutic dose of a vasopressor for at least 2 hours.

[0073] In a preferred embodiment, the composition is used to treat hypotension in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or acute lung injury, preferably persistent hypotension, preferably for treating hypotension in septic shock, preferably the persistent hypotension, wherein the hypotension, preferably the persistent hypotension, is associated with systolic blood pressure <90 mmHg or mean arterial pressure <70 mmHg, and wherein the hypotension, preferably the persistent hypotension, is pretreated with at least one, preferably a therapeutic dose of a vasopressor for at least 2 hours, wherein the vasopressor is selected from dopamine, adrenaline, noradrenaline, phenylephrine, or vasopressin.

[0074] In a preferred embodiment, the composition is used to treat hypotension in septic shock, sepsis, severe sepsis, acute respiratory distress syndrome, or acute lung injury, preferably persistent hypotension, preferably for treating hypotension in septic shock, preferably persistent hypotension, wherein the hypotension, preferably persistent hypotension, is associated with systolic blood pressure <90 mmHg or mean arterial pressure <70 mmHg, and wherein the hypotension, preferably persistent hypotension, is pretreated with at least one, preferably a therapeutic dose of a vasopressor for at least 2 hours, wherein the therapeutic dose of the vasopressor is >5 mg / kg / min of dopamine or a corresponding dose of the vasopressor, preferably a corresponding dose of adrenaline, noradrenaline, phenylephrine, or vasopressin.

[0075] In a preferred embodiment, the composition is used to treat and prevent septic shock. In a preferred embodiment, the composition is used to treat hypotension, preferably persistent hypotension. In a preferred embodiment, the hypotension, preferably persistent hypotension, is associated with septic shock. In a preferred embodiment, the composition is used to treat persistent hypotension. In a preferred embodiment, the persistent hypotension is associated with septic shock.

[0076] In a preferred embodiment, the hypotension, preferably persistent hypotension, is associated with a systolic blood pressure <90 mmHg (or mean arterial pressure <70 mmHg). In a preferred embodiment, the hypotension, preferably persistent hypotension, is associated with a systolic blood pressure <90 mmHg (or mean arterial pressure <70 mmHg) even after adequate fluid resuscitation with a therapeutic dose of a vasopressor (i.e., dopamine >5 mg / kg / min or any dose of epinephrine, noradrenaline, phenylephrine, or vasopressin) for at least 2 hours.

[0077] In a preferred embodiment, the treatment is adjunctive to antibiotic therapy, preferably adjunctive to standard antibiotic therapy. In a preferred embodiment, the antibiotic therapy, preferably the standard antibiotic therapy, comprises antibiotics selected from: ceftriaxone, spiramycin, amoxicillin, amoxicillin / clavulanic acid, gentamicin, piperacillin / tazobactam, cefuroxime, penicillin, azithromycin, clarithromycin, erythromycin, doxycycline, cefotaxime, ampicillin, ertapenem, cefepime, imipenem, meropenem, ciprofloxacin, levofloxacin, vancomycin, linezolid, moxifloxacin, and gemifloxacin, and wherein the antibiotic therapy, preferably the standard antibiotic therapy, comprises antibiotics selected from: ceftriaxone, spiramycin, amoxicillin, gentamicin, levofloxacin, piperacillin / tazobactam, amoxicillin / clavulanic acid, cefuroxime, and penicillin. In a preferred embodiment, the antibiotic therapy, preferably standard antibiotic therapy, is intravenous (IV) or oral antibiotic therapy, preferably standard antibiotic therapy.

[0078] In a preferred embodiment, the human patient has pneumonia, preferably selected from community-acquired pneumonia (CAP), hospital-acquired pneumonia (HAP), and ventilator-associated pneumonia (VAP). In a preferred embodiment, the human patient has pneumonia, wherein the pneumonia is community-acquired pneumonia (CAP). In a preferred embodiment, the human patient has pneumonia, wherein the pneumonia is hospital-acquired pneumonia (HAP). In a preferred embodiment, the human patient has pneumonia, wherein the pneumonia is ventilator-associated pneumonia (VAP). In a preferred embodiment, the human patient has pneumonia, wherein the pneumonia is severe pneumonia, preferably severe community-acquired pneumonia (sCAP) or severe community-acquired pneumococcal pneumonia (sCAPP). In a preferred embodiment, the human patient has pneumonia, wherein the pneumonia is community-acquired pneumonia (CAP) or community-acquired pneumococcal pneumonia (CAPP).

[0079] In a preferred embodiment, the human patient suffers from pneumonia, and the pneumonia is caused by: Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecium, Legionella pneumophila, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumanii, Bordetella pertussis, Serratia marcescens, Stenotrophomonas maltophilia, Moraxella catarrhalis, or Mycobacterium tuberculosis. In a preferred embodiment, the human patient suffers from pneumonia, and the pneumonia is severe pneumonia caused by: Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Legionella pneumophila, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Serratia marcescens, Stenotrophomonas maltophilia, Moraxella catarrhalis, or Mycobacterium tuberculosis, and preferably the pneumonia, preferably the severe pneumonia, is caused by: Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Serratia marcescens, or Mycobacterium tuberculosis, and further preferably the pneumonia, preferably the severe pneumonia, is caused by: Streptococcus pneumoniae, Staphylococcus aureus, Klebsiella pneumoniae, or Pseudomonas aeruginosa, and again, more preferably the pneumonia or the severe pneumonia is caused by Streptococcus pneumoniae.

[0080] In a preferred embodiment, the composition is in the form of a solution for intravenous administration, preferably intravenous infusion, wherein each liter of solution comprises a mixture of the empty liposomes between 10 g and 40 g, preferably between 10 g and 20 g per liter of solution.

[0081] In a preferred embodiment, the intravenous infusion time is up to 3 hours, and preferably the intravenous infusion time is from 10 minutes to 2 hours.

[0082] In a preferred embodiment, the composition is administered in at least two doses. In a preferred embodiment, the composition is administered in at least two doses: a first dose and a second dose, wherein the interval between the first dose and the second dose is 6 hours to 96 hours, preferably 12 hours to 72 hours, more preferably 24 hours to 48 hours, and even more preferably 24 hours or 48 hours.

[0083] In a preferred embodiment, the composition is administered in 2 to 4 doses, preferably two doses, over 12 to 72 hours, more preferably two doses over 24 to 48 hours, preferably at 24 or 48-hour intervals.

[0084] In a preferred embodiment, the composition is administered to the human patient in at least two doses: a first dose and a second dose, wherein the interval between the first dose and the second dose is 20 to 28 hours, preferably 24 hours.

[0085] In a preferred embodiment, the composition is administered to the human patient in two doses: a first dose and a second dose, wherein the administration of the first dose and the administration of the second dose are spaced 20 to 28 hours, preferably 24 hours apart.

[0086] In a preferred embodiment, each of the doses is from 1 mg / kg to 64 mg / kg, preferably from 2 mg / kg to 32 mg / kg, more preferably from 3 mg / kg to 25 mg / kg, and even more preferably from 4 mg / kg to 16 mg / kg.

[0087] In a preferred embodiment, each of the doses is from 2 mg / kg to 8 mg / kg, preferably from 2 mg / kg to 6 mg / kg, more preferably from 3 mg / kg to 5 mg / kg, and even more preferably from 4 mg / kg.

[0088] In a preferred embodiment, each of the doses is from 10 mg / kg to 22 mg / kg, preferably from 12 mg / kg to 20 mg / kg, more preferably from 14 mg / kg to 18 mg / kg, and even more preferably from 16 mg / kg.

[0089] In a preferred embodiment, the composition is administered in the form of a solution for intravenous administration.

[0090] In a preferred embodiment, the composition is administered to the human patient, preferably in at least two doses: a first dose and a second dose, wherein the administration of the first dose and the administration of the second dose are spaced 20 to 48 hours, preferably 24 hours apart, and wherein the composition is in the form of a solution for intravenous administration.

[0091] In a preferred embodiment, the composition is used to treat sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, and wherein the composition is preferably used to treat septic shock in animals, preferably humans, and wherein the sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, preferably septic shock requires hospitalization.

[0092] In a preferred embodiment, the composition is used to treat sepsis, septic shock, or hypotension in animals, preferably humans, and preferably persistent hypotension.

[0093] In a preferred embodiment, the composition is used to treat sepsis, septic shock, or hypotension in a person, preferably persistent hypotension. In a very preferred embodiment, the composition is used to treat sepsis in a person. In a very preferred embodiment, the composition is used to treat septic shock in a person. In a very preferred embodiment, the composition is used to treat hypotension in a person, preferably persistent hypotension. In a preferred embodiment, the sepsis, septic shock, or hypotension, preferably persistent hypotension, requires hospitalization, preferably in a hospital intensive care unit (ICU).

[0094] In a preferred embodiment, the composition is used to treat sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, and wherein the composition is preferably used to treat septic shock in animals, preferably humans, and wherein the sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, preferably septic shock requires hospitalization in an intensive care unit (ICU), preferably in a hospital's intensive care unit (ICU).

[0095] In a preferred embodiment, the composition is used to treat sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, and wherein the composition is preferably used to treat septic shock in animals, preferably humans, wherein the sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, preferably septic shock requires hospitalization, and wherein the treatment reduces the length of hospital stay compared to the length of hospital stay without such treatment.

[0096] In a preferred embodiment, the hospital stay is reduced by at least one day, preferably two days, more preferably three days, even more preferably four days, even more preferably five days, even more preferably six days, even more preferably seven days, even more preferably eight days, and even more preferably nine days due to the treatment. In a preferred embodiment, the hospital stay is at most 18 days.

[0097] In a preferred embodiment, the composition is used to treat sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, and wherein the composition is preferably used to treat septic shock in animals, preferably humans, wherein the sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, preferably septic shock requires hospitalization in an intensive care unit (ICU), preferably in a hospital intensive care unit (ICU), and wherein the treatment reduces the length of hospitalization in the intensive care unit (ICU) compared to the length of hospitalization in the intensive care unit (ICU) without such treatment.

[0098] In a preferred embodiment, the length of stay in the intensive care unit (ICU) is reduced to at least one day, preferably two days, more preferably three days, even more preferably four days, even more preferably five days, even more preferably six days, and even more preferably seven days. In a preferred embodiment, the length of stay in the intensive care unit (ICU) is at most 18 days.

[0099] In a preferred embodiment, the composition is used to treat sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, and wherein the composition is preferably used to treat septic shock in animals, preferably humans, and wherein the sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, preferably septic shock, is cured in less time than when such treatment is not performed.

[0100] In a preferred embodiment, the cure time is less than one day, preferably less than two days, or more preferably less than three days, even more preferably less than four days, even more preferably less than five days, even more preferably less than six days, and even more preferably less than seven days.

[0101] In a preferred embodiment, the composition is used to treat sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, and wherein the composition is preferably used to treat septic shock in animals, preferably humans, and wherein the treatment reduces the cardiovascular SOFA score compared to when such treatment was not performed.

[0102] In a preferred embodiment, the composition is used to treat sepsis, severe sepsis, septic shock, or prolonged and severe hypotension, preferably persistent hypotension, and wherein the composition is preferably used to treat septic shock in animals, preferably humans, and wherein the treatment reduces the cardiovascular SOFA score compared to when such treatment was not performed, and wherein the reduction is at least 50%, preferably at least 60%, further preferably at least 70%, and even more preferably at least 80% after 7 days of treatment initiation, preferably 6 days of treatment initiation, more preferably 5 days of treatment initiation.

[0103] Example

[0104] Liposomes:

[0105] Sphingomyelin from egg yolk (CAS No.: 85187-10-6) was purchased from Sigma (S0756), Avanti Polar Lipids (860061), or Lipoid GmbH. Cholesterol from sheep lanolin (CAS No.: 57-88-5) was purchased from Sigma (C-8667), Avanti Polar Lipids (70000), or Dishman Netherlands B.V. According to the invention, the sphingomyelin and cholesterol included in or composed of the mixture of empty liposomes of the invention can be obtained from natural sources as described above, or alternatively obtained through chemical synthesis.

[0106] Liposome preparation:

[0107] Single-layer sphingomyelin: cholesterol (molar ratio of 35:65) and sphingomyelin-only (100%) liposomes were prepared by ultrasonic treatment or microfluidization (e.g., high-pressure homogenization) or according to hydration, extrusion and percolation process specifications.

[0108] Ultrasound:

[0109] Lipids were dissolved in chloroform at a concentration of 1 mg / ml and stored at -20°C. To prepare liposomes, the chloroform solutions of each lipid were mixed in proportion as needed to routinely produce a final solution of 50 μl to 500 μl. The chloroform was completely evaporated at 60°C for 20 to 50 minutes. 50 μl or 100 μl of Tyrode's buffer (140 mM NaCl, 5 mM KCl, 1 mM MgCl2, 10 mM glucose, 10 mM HEPES; pH 7.4) containing 2.5 mM CaCl2 was added to a tube containing the dried lipid membrane and vortexed vigorously. The lipid suspension was incubated in an Eppendorf hot mixer at 45°C for 20 to 30 minutes with vigorous shaking. To generate liposomes, the final lipid suspension was sonicated in a Bandelin Sonopuls sonicator at 70% power at 6°C for 3 × 5 seconds. Before using the liposome formulation in experiments, place it at 6°C for at least 1 hour.

[0110] Hydration, extrusion and diafiltration procedure:

[0111] In alternative methods, each liposome formulation is prepared via ethanol hydration and extrusion. The lipids are dissolved separately in ethanol and tert-butanol while being mixed at high temperature (~55°C). The lipid solution is then added to a PBS buffer solution (sodium chloride, monosodium phosphate dihydrate, and disodium phosphate dihydrate dissolved in water for injection, adjusted to pH 7.0 to 7.4 with hydrochloric acid (HCl) or sodium hydroxide (NaOH) as needed, and filtered through a 0.2 μm filter) while mixing at high temperature (~65°C) for approximately 30 minutes. The resulting process fluid is then repeatedly extruded through a series of polycarbonate track-etched films under high pressure and high temperature (~65°C) until the desired particle size, as measured by dynamic light scattering, is achieved (example of an extruder:). (Extruder). The resulting process fluid was then concentrated approximately 2-fold using a hollow fiber filter cartridge with a molecular weight cutoff of 100,000, and then percolated against approximately 10-fold volume exchange with PBS buffer to remove ethanol and n-butanol. At the end of percolation, the process fluid was concentrated approximately 30% to allow for subsequent dilution to the target lipid concentration. Prior to dilution, the process fluid was filtered through a 0.2 μm sterile-grade filter to remove any larger liposomes that might clog the filter during aseptic filtration. The process fluid was then diluted with PBS buffer to the target total lipid concentration of 40 mg / mL. The final formulation was aseptically filtered through two tandem 0.2 μm sterile-grade filters and aseptically filled into glass tubular vials.

[0112] The concentration of individual lipids in liposomes is always given as a weight-to-weight ratio. In liposomes containing sphingomyelin and cholesterol, a 1:1 (weight-to-weight) ratio corresponds to a 50% (weight-to-weight) or 35:65 molar ratio. Specifications are described in Table 1.

[0113] Table 1: Liposome specifications

[0114] Mean diameter (nm) Polydispersity index Zeta potential (mV) Osmolality (mmol / kg) pH 40 to 400 <0.45 -25 to +2 250-400 6.5-8.0

[0115] CURB-65 score:

[0116] CURB-65 (also known as the CURB criteria) is a well-established clinical predictive rule of thumb for community-acquired pneumonia mortality, well-known to those skilled in the art (Lim WS et al. (2003), Thorac 58(5):377–82). The British Thoracic Society recommends CURB-65 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". Thorac 56. Supplement 4:IV1–64).

[0117] The rating is an acronym for each of the measured risk factors. Each risk factor is rated on a scale of one point, with a maximum rating of 5.

[0118] - New-onset confusion (defined as 8 or fewer AMTS)

[0119] - Blood urea nitrogen greater than 7 mmol / L (19 mg / dL)

[0120] - Breathing rate of 30 breaths per minute or more

[0121] - Blood pressure less than 90 mmHg, systolic or diastolic blood pressure of 60 mmHg or less.

[0122] - Must be 65 years of age or older.

[0123] APACHE II:

[0124] 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., October 1985; 13(10):818-29).

[0125] Table 2: APACHE II rating

[0126]

[0127]

[0128]

[0129]

[0130] The APACHE II score aims to provide an approximation of the likelihood of death for a patient or a group of patients based on the total score obtained. Table 3 describes the interpretation of the score for mortality.

[0131] Table 3: Explanation of Apache II Score / Approximate Mortality Rate

[0132]

[0133] SOFA:

[0134] SOFA score is an acronym for Sequential Organ Failure Assessment, and is calculated according to Table 4 (S. Vosylius, J. Sipylaite and J. Ivaskevicius, Croatian Medical Journal, 45(2004), 715-20):

[0135] Table 4: SOFA rating

[0136]

[0137]

[0138] Cardiovascular hypotension:

[0139] Cardiovascular hypotension is a major characteristic of septic shock. It may be due to low cardiac output or low systemic vascular resistance. Its severity can be assessed using the cardiovascular SOFA (Sequential Organ Failure Assessment) score, which is defined by mean arterial pressure or by the need for vasopressors, as shown in Table 5 (S. Vosylius, J. Sipylaite and J. Ivaskevicius, Croatian Medical Journal, 45(2004), 715-20).

[0140] Table 5: Cardiovascular SOFA score

[0141] Mean arterial pressure or need for vasopressor administration Score No hypotension 0 MAP < 70 mm / Hg 1 Dopamine < 5 pg / kg / min or dobutamine (any dose) 2 Dopamine > 5 pg / kg / min or epinephrine < 0.1 pg / kg / min or norepinephrine < 0.1 pg / kg / min 3 Dopamine > 15 pg / kg / min or epinephrine > 0.1 pg / kg / min or norepinephrine > 0.1 pg / kg / min 4

[0142] Example 1

[0143] Treatment of sepsis and septic shock in patients with severe infections

[0144] In a subsequently reported study, a highly preferred mixture of empty liposomes of the present invention (designated CAL02) was administered intravenously (IV) as adjunctive therapy in addition to standard antibiotic therapy to patients admitted to the intensive care unit (ICU) with severe community-acquired pneumonia (CAP) caused by Streptococcus pneumoniae. The highly preferred mixture of empty liposomes of the invention (CAL02) comprises a 1:1 (weight / weight-w / w) mixture of a first empty liposome and a second liposome, wherein the first empty liposome consists of sphingomyelin and cholesterol in a 1:1 weight ratio (1:1 w / w; molar ratio 35:65), and the second empty liposome consists only of sphingomyelin.

[0145] In addition to standard-of-care antibiotic therapy (Mandel et al., CID 2007:44(Supplement 2), S27-S72), each patient received two infusions of CAL02 or placebo (physiological 0.9% NaCl solution) (with intervals of 24 or 48 hours between these administrations). The first administration was given shortly after the severity was diagnosed. The diagnosis of severity was based on at least one of the following severity criteria:

[0146] i. Invasive mechanical ventilation support

[0147] ii. After adequate fluid resuscitation, treat with a therapeutic dose of vasopressor (i.e., dopamine >5 mg / kg / min or any dose of epinephrine, norepinephrine, phenylephrine or vasopressin) for at least 2 hours to maintain or attempt to maintain systolic blood pressure >90 mm Hg (or mean arterial pressure >70 mm Hg).

[0148] Or based on at least three of the following minor severity criteria:

[0149] i. Respiratory rate ≥30 breaths / minute

[0150] ii. PaO2 / FiO2 ratio ≤ 250 mm Hg

[0151] iii. Multi-leaved infiltration

[0152] iv. Confusion / disorientation (must be documented before use of sedatives or other new psychotropic drugs)

[0153] v. Urea >7mM (>40mg / dL)

[0154] vi. Leukopenia (white blood cell count < 4,000 cells / mm²) 3 )

[0155] vii. Thrombocytopenia (platelet count <100,000 cells / mm) 3 )

[0156] viii. Hypothermia (core temperature <36℃)

[0157] ix. Systolic blood pressure <90 mm Hg or mean arterial pressure <70 mm Hg, and received fluid resuscitation of ≥40 mL / kg for at least 2 hours.

[0158] Two dose levels of CAL02 (4 mg / kg (low dose) and 16 mg / kg (high dose)) were tested. Five patients were randomly assigned to the placebo group, 11 patients were assigned to the high-dose CAL02 group, and 3 patients were assigned to the low-dose CAL02 group.

[0159] At the time of treatment, 56% of the patients had septic shock: 2 patients in the placebo group, 5 patients in the high-dose CAL02 group, and all three patients in the low-dose CAL02 group.

[0160] Study results

[0161] Table 6: Patient characteristics at baseline

[0162]

[0163] When considering the entire study population and when assessing only patients presenting with septic shock at baseline, a faster decline in the cardiovascular SOFA score was observed in the CAL02 group compared to the placebo group: in the CAL02 group, the SOFA score decreased by 100% within 6 days, while at the same time point, the placebo group did not reach a 40% reduction (Figures 1 and 2).

[0164] In patients presenting with septic shock, on day 8 after treatment, hypotension and septic shock completely resolved in all patients (5 / 5, 100%) in the high-dose CAL02 group and in 66% (2 / 3) of patients in the low-dose CAL02 group, compared to no patients (0 / 2, 0%) in the placebo group.

[0165] In the placebo group, of the three patients who were not in septic shock at baseline, one patient had hypotension at baseline and developed septic shock on day 4, and two patients did not have hypotension at baseline but developed hypotension within the first 8 days. In contrast, in the CAL02 group, three patients did not have hypotension at baseline and did not develop any hypotension, and all other patients improved to a remission state by day 6. This indicates that CAL02 prevents hypotension and hemodynamic instability and prevents the occurrence of septic shock.

[0166] The resolution of septic shock was accompanied by a significant reduction in the mean ICU stay, from 32 days in the placebo group to 5.4 days and 15 days in the high-dose CAL02 group and the low-dose CAL02 group, respectively. Furthermore, the mortality rate was lower in the CAL02 group compared to the placebo group (50%) (20% and 33% mortality rates in the high-dose and low-dose CAL02 groups, respectively) (Table 7).

[0167] Table 7: Results

[0168]

[0169] The effects of CAL02 treatment on vital signs (including heart rate, systolic and diastolic blood pressure, and core body temperature) and lactate levels were also evaluated.

Claims

1. A composition comprising a mixture of empty liposomes, preferably consisting of said mixture of empty liposomes, wherein said mixture of empty liposomes comprises, preferably, the following: (a) A first empty liposome containing cholesterol, wherein the amount of cholesterol is at least 30% (by weight); as well as (b) A second empty liposome comprising sphingomyelin, wherein preferably the second empty liposome (b) comprises sphingomyelin as the sole lipid component; The composition is used to treat sepsis in animals, preferably humans.

2. The composition for use according to claim 1, wherein the composition is used to treat septic shock in the animal, preferably the human.

3. The composition for use according to claim 1, wherein the composition is used to treat hypotension, preferably persistent hypotension, in the animal, preferably the human, suffering from sepsis.

4. The composition for use according to claim 2, wherein the composition is used to treat hypotension, preferably persistent hypotension, in the animal, preferably the human, suffering from septic shock.

5. The composition for use according to any one of the preceding claims, wherein the amount of cholesterol in the empty liposome (a) is 45% to 55% (by weight), and wherein the second empty liposome (b) is composed of sphingomyelin.

6. The composition for use according to any one of the preceding claims, wherein the first empty liposome (a) is composed of cholesterol and sphingomyelin, and wherein the amount of cholesterol in the empty liposome (a) is about 50% (w / w), and wherein the mixture of the empty liposomes comprises at least 40%, preferably at least 45% (w / w) of the first empty liposome (a) and the second empty liposome (b).

7. The composition for use according to any one of the preceding claims, wherein the first empty liposome (a) comprises a 1:1 (weight / weight-w / w) mixture of the first empty liposome and the second liposome, wherein the first empty liposome is composed of cholesterol and sphingomyelin in a 1:1 weight ratio (1:1 w / w; molar ratio 35:65), and the second empty liposome is composed of sphingomyelin alone, wherein the first empty liposome (a) comprises the cholesterol and the sphingomyelin as the sole lipid component, and the second empty liposome (b) comprises the sphingomyelin as the sole lipid component.

8. The composition for use according to any one of claims 3 to 7, wherein the hypotension, preferably the persistent hypotension, is associated with a mean arterial pressure <70 mm Hg.

9. The composition for use according to claim 8, wherein the hypotension, preferably the persistent hypotension, is pretreated with a vasopressor for at least 2 hours.

10. The composition for use according to any one of the preceding claims, wherein the treatment is an adjunct to antibiotic therapy.

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

12. The composition for use according to any one of the preceding claims, wherein the animal is a human patient, and wherein the human patient has pneumonia, wherein preferably the pneumonia is selected from community-acquired pneumonia (CAP), hospital-acquired pneumonia (HAP), and ventilator-associated pneumonia (VAP).

13. The composition for use according to claim 12, wherein the pneumonia is severe pneumonia, preferably severe community-acquired pneumonia (sCAP) or severe community-acquired pneumococcal pneumonia (sCAPP).

14. The composition for use according to claim 12 or 13, wherein the pneumonia or the severe pneumonia is caused by: Streptococcus pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Legionella pneumophila, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Bordetella pertussis, Serratia marcescens, Stenotrophomonas maltophilia, Moraxella catarrhalis, or Mycobacterium tuberculosis, wherein preferably the pneumonia or the severe pneumonia is caused by Streptococcus pneumoniae.

15. The composition for use according to any one of the preceding claims, wherein the composition is in the form of a solution for intravenous administration.

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

17. The composition for use according to any one of the preceding claims, wherein the sepsis, septic shock, or the hypotension, preferably persistent hypotension, requires hospitalization, preferably in a hospital intensive care unit (ICU).

18. The composition for use according to claim 17, wherein the treatment reduces the length of hospital stay in the hospital compared to the length of hospital stay without such treatment.

19. The composition for use according to claim 17 or claim 18, wherein the reduction in hospitalization time due to the treatment is at least one day, preferably two days, more preferably three days, even more preferably four days, even more preferably five days, even more preferably six days, even more preferably seven days, even more preferably eight days, even more preferably nine days.

20. The composition for use according to any one of the preceding claims, wherein the sepsis, septic shock, or hypotension, preferably persistent hypotension, takes less time to heal compared to the absence of such treatment.

21. The composition for use according to claim 20, wherein the cure time is less than one day, preferably less than two days, or more preferably less than three days, even more preferably less than four days, even more preferably less than five days, even more preferably less than six days, and even more preferably less than seven days.

22. The composition for use according to any one of the preceding claims, wherein the treatment reduces the cardiovascular SOFA score compared to the cardiovascular SOFA score without such treatment.

23. The composition for use according to claim 22, wherein the reduction is at least 50%, preferably at least 60%, more preferably at least 70%, and even more preferably at least 80% seven days after the start of treatment, preferably six days after the start of treatment, and even more preferably five days after the start of treatment.

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