Compositions and methods for treatment of hypercalcitonemia

Abstract

Disclosed is a composition for the treatment of hypercalcitonemia, the composition comprising a lipophilic or hydrophobic component, an amphiphilic emulsifier, a polar liquid carrier, and comprising or without one or more electrolytes wherein the amphiphilic emulsifier forms: a micelle in a polar liquid carrier, the micelle is provided with a lipophilic or hydrophobic core containing a lipophilic or hydrophobic component; and / or self-assembling to form lipidosome with a lipid bilayer structure; and / or other particle configurations.

Description

[0001] Related Applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 482,540, filed January 31, 2023. TECHNICAL FIELD

[0003] The present application relates generally to pharmaceutical compositions, and more particularly to compositions for treating hyperprocalcitoninemia. BACKGROUND

[0004] Procalcitonin is a peptide composed of 116 amino acids with a molecular weight of approximately 13.0 kDa. Procalcitonin is expressed in the parafollicular cells of the thyroid gland and is enzymatically cleaved therein to yield the calcium-regulating hormone calcitonin. Normally, procalcitonin production is restricted to these cells, and its concentration in the blood is extremely low, less than 0.05 ng / ml. Under conditions of severe physiological stress, monocytes and parenchymal cells of many organs such as fat, liver, lung, muscle, stomach, kidney, and brain can synthesize procalcitonin, leading to a 100,000-fold increase in its blood concentration. The enzyme that converts procalcitonin to calcitonin is lacking in tissues other than the thyroid. This leads to the release of procalcitonin into the blood instead of calcitonin during physiological stress. Recent studies have shown that procalcitonin is not only a marker of the inflammatory response but can also act as a mediator of the inflammatory response, thus becoming a target for the treatment of diseases in which its concentration is elevated.

[0005] The inflammatory response is the pathophysiological basis of a variety of disorders. These diseases can or can not involve infection. Examples include bacterial or viral infection, malaria, blood loss, severe heart failure, brain and / or spinal cord injury, bone fracture, non-orthopedic trauma, pancreatitis, burns, ischemic stroke, subarachnoid hemorrhage, heat stroke, vasculitis, metastatic cancer, dementia, sarcopenia, aging, chronic or acute kidney disease, chronic or acute liver failure, heart failure, acute respiratory distress syndrome, and arthritis. In vitro and in vivo studies have revealed the role of procalcitonin as a mediator of inflammation and tissue damage, which are characteristic of numerous such disorders.

[0006] Procalcitonin has been shown to increase intracellular concentrations of calcium, similar to the proinflammatory cytokine interleukin 8 (Becker et al., “Procalcitonin in sepsis and systemic inflammation: a harmful biomarker and therapeutic target,” British Journal of Pharmacology, 2010, vol. 159, no. 2: pp. 253-264).

[0007] Studies on rat aortic vascular smooth muscle cells have shown that procalcitonin does not directly affect the release of nitric oxide from these cells. However, when used in combination with lipopolysaccharide, tumor necrosis factor and interferon gamma, procalcitonin significantly increases the production of nitric oxide from these cells (Tilg, Peschel, “Interferon-alpha and its effects on the cytokine cascade: pro- and anti-inflammatory cytokines”, Leukemia, 1996, vol. 23, no. 1-2: 55-60). Procalcitonin is thus not a primary inducer, but an amplifier of the inflammatory response. Moreover, procalcitonin has direct effects, making it a toxic mediator.

[0008] The literature is rich in examples of procalcitonin as a toxic mediator. Wagner et al. (Wagner et al., “Procalcitonin impairs endothelial cell function and viability”, Am J Physiol Heart Circ Physiol, 2017, vol. 124, no. 3: 835-845) have shown that procalcitonin induces endothelial barrier disruption, cell migration, neocapillary formation and endothelial cell death. In addition, the recovery of perfusion of the hind limb of the ischemic mouse leg is impaired. Sauer et al. (Sauer et al., “Procalcitonin impairs hepatocyte viability and function in vitro: a potential new mechanism for liver dysfunction and failure during sepsis?”, Int J Bio Med Res, 2017, vol.: Open Access article 6130725) found that procalcitonin inhibits hepatocyte proliferation and induces hepatocyte death. The role of procalcitonin as a toxic mediator is further demonstrated by the use of procalcitonin-deficient mice. These mice are protected from septic shock and exhibit reduced lung inflammation (Baranowsky et al., “Procalcitonin acts as a mediator in septic shock via the calcitonin gene-related peptide receptor”, Crit Care, 2021, vol. 49 (e41-e52)). Sullivan and Schmidt elucidate the impairment of microcirculation by procalcitonin (Sullivan and Schmidt, “Procalcitonin: a mediator of microvascular dysfunction during critical illness”, Am J Respir Crit Care Med, 2022, vol. 206, no. 4: 375-376). Infusion of procalcitonin into septic animals increases mortality, while administration of procalcitonin antibodies reduces mortality in septic animals (Becker et al., “Procalcitonin in sepsis and systemic inflammation: a harmful biomarker and therapeutic target”, Br J Pharmacol, 2010, vol. 159, no. 2: 253-264).

[0009] In humans, lower blood procalcitonin concentrations have been shown to be indicative of a good prognosis for sepsis, acute respiratory distress syndrome, fractures, acute kidney injury, heart failure and trauma. Given its role as an amplifier of the inflammatory response and the direct adverse effects of procalcitonin, drugs that reduce procalcitonin have a benefit in acute or chronic conditions in which procalcitonin is elevated. SUMMARY

[0010] In one aspect, the present application relates to a method for treating hyperprocalcitoninemia in a subject in need thereof. The method comprises the following steps: administering to the subject in need thereof an effective amount of a nanoparticle composition comprising: 0.1%-100% (w / v) of an amphiphilic emulsifier; 0-35% (w / v) of a lipophilic or hydrophobic component; a polar liquid carrier; and one or more electrolytes, wherein the nanoparticle composition comprises liposomes and / or micelles having an average particle size of 1-800 nm.

[0011] Another aspect of the present application relates to a method for treating a disease or condition associated with elevated procalcitonin in a subject, including but not limited to chronic or acute renal failure, chronic or acute liver failure, chronic or acute respiratory failure, sarcopenia in the elderly, dementia, orthopedic or non-orthopedic trauma, surgery, myocardial infarction, autism, ischemic stroke, Parkinson's disease, vasculitis, bone fracture, blood loss, and depression. The method comprises the following steps: administering to a subject in need of treatment for hyperprocalcitoninemia an effective amount of a hydrophobic nanoparticle composition, the composition comprising: 0.1%-100% (w / v) of an amphiphilic emulsifier; 0-35% (w / v) of a lipophilic or hydrophobic component; a polar liquid carrier; and one or more electrolytes, wherein the nanoparticle composition comprises liposomes and / or micelles having a particle size range of 1-800 nm.

[0012] Another aspect of the present application relates to a method for treating aging, comprising administering to a subject in need of treatment an effective amount of a nanoparticle composition comprising: 0.1%-100% (w / v) of an amphiphilic emulsifier; 0-35% (w / v) of a lipophilic or hydrophobic component; a polar liquid carrier; and one or more electrolytes, wherein the nanoparticle composition comprises liposomes and / or micelles having a particle size range of 1-800 nm.

[0013] Detailed description

[0014] Below with reference to certain aspects and exemplary embodiments of the present application, the technical structure in the specification and the example in the accompanying drawings are elaborated in detail. Various aspects of the application will be described in conjunction with exemplary embodiments (including methods, materials and examples), and such description is non-restrictive, and the scope of the application is intended to cover all known or equivalents, alternatives and modifications included in the application. Unless otherwise defined, the meaning of all technical and scientific terms used in this application is consistent with that commonly understood by those of ordinary skill in the art to which this application belongs. Those skilled in the art will recognize that a variety of similar or equivalent techniques and materials described in this application can be used to practice various aspects and embodiments of the application. Various aspects and embodiments described in this application are not limited to described methods and materials.

[0015] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0016] Ranges can be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such ranges are expressed, another embodiment includes from the one particular value and to the other particular value. Similarly, when values are expressed as approximations by using the term "about," it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are not significant

[0017] I. DEFINITIONS

[0018] The term "acute critical illness" refers to any condition that requires immediate intensive care treatment of a patient. Such conditions can be triggered by any acute and extensive damage suffered by the body, including but not limited to physical trauma, burn trauma, infection (under which are included sepsis, severe sepsis, blood loss), systemic inflammatory response syndrome (SIRS), acute myocardial infarction, diabetic ketoacidosis or other thromboembolic events.

[0019] The term "chronic illness" refers to any condition that does not require immediate treatment, whose effects persist or exist for a long time or progress over time. Such conditions can be triggered by any long-standing disease, including but not limited to arthritis, chronic renal failure, liver failure, cancer metastasis, dementia, Parkinson's disease or schizophrenia.

[0020] The term "intensive care treatment", also referred to herein as "organ support care", can include, but is not limited to, ventilation therapy (e.g. mechanical ventilation), hemodialysis, vasopressor therapy, fluid therapy, transfusion therapy with administration of red blood cell concentrates / fresh frozen plasma / platelet concentrates / whole blood / coagulation factor concentrates, systemic antibiotic and / or antiviral and / or antifungal and / or antiprotozoal therapy, parenteral nutrition, granulocyte transfusion, T cell transfusion, stem cell transfusion, anticoagulation and / or antithrombotic therapy (including low molecular weight heparin), corticosteroid administration, strict glycemic control, and therapy by ventricular assist devices, intra-aortic balloon counterpulsation pumps, plasmapheresis, extracorporeal membrane oxygenation (ECMO) and rotary mechanical circulatory support systems, etc.

[0021] The term "trauma" as used herein refers to any impact injury or physical injury caused by an external factor such as blast trauma, crush injury, blunt trauma, penetrating trauma, chemical injury (leak, war or poisoning), radiation or burns, etc. to living tissue, injury or accident.

[0022] The term "hypercalcitoninemia" as used herein refers to a condition in which the level of calcitonin in the blood of a human is higher than normal. In some embodiments, hypercalcitoninemia refers to a condition in which the level of calcitonin in the blood of a human is at or above 0.05 ng / ml.

[0023] The term "treatment" as used herein refers to the management and care of a patient having hypercalcitoninemia or at risk of developing hypercalcitoninemia. The term is intended to include the full spectrum of treatments for the condition, e.g., the administration of the phospholipid nanoparticle composition of the present application for the purpose of effecting an improvement in, reducing or alleviating a symptom or complication; delaying the progression of the condition, disease or disorder; curing or eliminating the condition, disease or disorder; and / or reducing the risk of or preventing the condition, disease or disorder, including preventing the recurrence of the disease, wherein "prevention" is understood to mean the management and care of a patient for the purpose of preventing the development of the condition, disease or disorder, and includes the administration of the nanoparticle composition to prevent the occurrence of a symptom or complication. The individual to be treated is a human or non-human animal. The individual to be treated according to the present application can be of different ages and / or genders.

[0024] The term "organ failure" refers to an alteration in organ function in a critically ill patient requiring medical intervention to achieve homeostasis and / or loss of compensatory function of the failing organ. The organs include, but are not limited to, the heart and blood vessels (heart failure, vasoplegia), the lungs (respiratory failure), the liver (liver failure), the kidneys (renal failure), the brain (encephalopathy), the skeleton (arthritis), or the brain (dementia, encephalitis or meningitis).

[0025] The term "sepsis" takes its conventional clinical meaning to refer to a state of systemic inflammation (called systemic inflammatory response syndrome (SIRS)) and the presence of a confirmed or suspected infection. "Severe sepsis" is defined as sepsis-induced organ dysfunction or tissue hypoperfusion (e.g., manifested by elevated calcitonin, tumor necrosis factor or interleukin-1, vasoplegia, elevated lactate, decreased urine output, or altered mental status). Sepsis can lead to severe sepsis, multiple organ dysfunction syndrome / multiple organ failure (MODS) and death.

[0026] The term "systemic inflammatory response syndrome" or "SIRS" is used in its conventional clinical sense to refer to a systemic inflammatory response to injury from an unrecognized infectious process. SIRS is diagnosed when at least two of the following criteria are present: 1) a body temperature below 36°C (96.8°F) or above 38°C (100.4°F); 2) a heart rate above 90 beats / minute; 3) tachypnea (high respiratory rate) above 20 breaths / minute or an arterial carbon dioxide partial pressure below 4.3 kPa (32 mmHg); and 4) a white blood cell count below 4000 cells / mm 3 (4×10 9 pcs / L) or more than 12,000 pcs / mm 3 (12×10 9 Sepsis is defined as suspected or confirmed (by culture, staining, or polymerase chain reaction (PCR)) with SIRS when infection is suspected or confirmed. Hyperprocalcitoninemia can be seen in sepsis (Schuetz et al., “Serial procalcitonin predicts mortality in patients with severe sepsis: results of the Multicenter Procalcitonin Monitoring Sepsis (MOSES) Study,” Critical Care Medicine, Vol. 20, No. 30, 2017: Open Access), severe blood loss (“Procalcitonin Release Patterns in Baboon Trauma and Sepsis Models: Relationships with Cytokines and Neopterin,” Critical Care Medicine, Vol. 28, No. 11, 2000: 3659–3663), and severe heart failure (Picariello et al., “Procalcitonin in Patients with Acute Coronary Syndrome and Cardiogenic Shock Undergoing Percutaneous Coronary Intervention,” Internal and Emergency Medicine, Vol. 4, No. 5, 2009: 403–408). Interleukin-6 has been shown to increase after brain and spinal cord injury, as well as during status epilepticus (Abdulla et al., "Neurogenic Inflammatory Response Syndrome: Role in Neurocritically Ill Patients," Journal of Neurocritical Care, Vol. 5, No. 2, 2022: 39-43). Thus, physiological disturbances caused by infection, blood loss, heart failure, or neurological injury differ in their triggering mechanisms. However, once triggered, they all share a common inflammatory component that leads to procalcitonin production.

[0027] The term "systemic inflammation" refers to alterations in organ function in acute or chronic patients resulting from the body's (vasculature, immune system, tissues) nonspecific conservative response to infection, noninfectious antigens, trauma, burns, organ / tissue destruction / degeneration / injury, ischemia, hemorrhage, intoxication, and / or malignancy.

[0028] The terms "micelle" and "lipid micelle (LM)" are used interchangeably herein to refer to a molecular aggregate dispersed in a liquid, comprising a hydrophilic "head" region in contact with the surrounding solvent, isolating a hydrophobic single-tail region in the center of the micelle, which forms a hydrophobic inner core suitable for accommodating and delivering hydrophobic drugs.

[0029] The term "liposome" as used herein refers to a vesicular structure composed of lipids, said lipids having tail groups comprising long hydrophobic hydrocarbon chains and hydrophilic head groups. The lipids are arranged to form a lipid bilayer, which has an aqueous inner core environment suitable for accommodating and delivering aqueous drugs and a lipid wall suitable for accommodating hydrophobic drugs, particularly gases such as oxygen.

[0030] The term "hydrophobic particle" includes micelles and liposomes, but also any particle that creates a hydrophobic space or accommodates hydrophobic substances, such as a hydrogel or a polycyclic compound (i.e. molecular cage) having a cavity for encapsulating hydrophobic substances.

[0031] II. Methods of treatment

[0032] One aspect of the present application relates to a method of treating or preventing hyperprocalcitonemia in a patient, the method comprising administering to the subject an effective amount of the nanoparticle composition of the present application. The inventors of the present application have surprisingly found that administration of the nanoparticle composition of the present application can prevent or reduce hyperprocalcitonemia.

[0033] Based on the body of knowledge in the art, infusion of hydrophobic nanoparticle compositions should increase procalcitonin. However, we found that it presents a large and sustained decrease. Procalcitonin is a peptide composed of 116 amino acids with a molecular weight of approximately 13.0 kDa. Procalcitonin is expressed in the parafollicular cells of the thyroid and is enzymatically cleaved there to produce the calcium-regulating hormone calcitonin. Normally, the production of procalcitonin is limited and its concentration in blood is very low, less than 0.05 ng / ml. Except for the thyroid, other tissues lack the enzyme that converts procalcitonin to calcitonin. But under severe physiological stress conditions, mononuclear cells and parenchymal cells of many organs can synthesize procalcitonin, causing its blood concentration to increase 100,000-fold. Procalcitonin elevation can be found in a variety of conditions, such as sepsis, acute renal failure, chronic renal failure, Alzheimer’s disease, vascular dementia, Lewy body dementia, frontotemporal dementia, encephalitis, meningitis, acute respiratory distress syndrome, COVID-19 infection, preeclampsia, lung cancer, colorectal cancer, ovarian cancer, heart failure, severe blood loss, and trauma. Procalcitonin has been considered a marker of disease severity. But it has also been demonstrated that procalcitonin itself has damaging effects. Procalcitonin elevation is derived from the increase of proinflammatory proteins such as tumor necrosis factor and interleukin 6 (Whang et al., “Interaction of Procalcitonin with Proinflammatory Cytokines in Sepsis,” Shock, vol. 14, no. 1, 2000, pp. 73-78; Nijsten et al., “Procalcitonin Behaves as a Rapidly Responding Acute-Phase Protein in Vitro and in Vivo,” Intensive Care Med., vol. 28, no. 2, 2000, pp. 458-461). Cai et al. (Inflamm. Med., 2010, Open Access Article ID 642462) demonstrated that both sepsis and severe blood loss induce tumor necrosis factor and high mobility group box 1 protein. Moreover, Reidl et al. (Intensive Care Med., vol. 28, no. 11, 2000, pp. 3659-3663) demonstrated that procalcitonin increases in both sepsis and severe blood loss.

[0034] Furthermore, Hierholzer et al. (Am. J. Physiol. 275(3):L611-621, 1998) demonstrated that severe blood loss results in increased interleukin-6. Infusion of hydrophobic nanoparticles consisting of liposomes and micelles (e.g., fat emulsions) has been shown to increase blood concentrations of proinflammatory mediators. Krough-Madesen et al. (Am. J. Physiol. Endocrinol. Metab. 294(2):E371-379, 2008) demonstrated that fat emulsions increase the production of proinflammatory mediators such as tumor necrosis factor and interleukin-6 after endotoxin infusion. Lou et al. (Mol. Nutr. Food Res. 65(5):2021) found that fat emulsion infusion increases multiple proinflammatory mediators including interleukin-6. Duncan et al. (FASEB J. 33(51):2019) demonstrated that infusion of fat emulsions into pregnant rats increases proinflammatory mediators. The increase in proinflammatory mediators caused by fat emulsions would lead one skilled in the art to expect that infusion of the nanoparticle compositions of the present application would result in an increase in procalcitonin. Furthermore, because the nanoparticle compositions would increase proinflammatory mediators, one skilled in the art would not administer the nanoparticle compositions of the present application to a patient in anticipation of decreasing procalcitonin and its deleterious effects. However, contrary to the expectations established by the literature, we found that procalcitonin continued to decrease after infusion of the nanoparticle compositions of the present application. It is also noteworthy that no serious adverse effects have been observed from the nanoparticle compositions of the present application. Thus, the unexpected decrease in procalcitonin (which itself promotes organ damage) by the nanoparticle compositions of the present application has important therapeutic applicability.

[0035] The nanoparticle compositions of the present application can be used to treat or prevent hypercalcitoninemia caused by a variety of different disease conditions triggered by tissue damage including, but not limited to, sepsis, major trauma, burns, pancreatitis, aspiration syndrome, extracorporeal circulation (e.g., heart bypass surgery), multiple transfusions, ischemia-reperfusion injury, autoimmune diseases, severe blood loss, heat-induced diseases, eclampsia, toxic / toxicity reactions. In some embodiments, the nanoparticle compositions of the present application are used to treat or prevent hypercalcitoninemia caused by sepsis triggered by infection with influenza virus or coronavirus (including SARS, MERS, and COVID-19 viruses).

[0036] Chronic pro-inflammatory state is a universal feature of aging. This chronic low-grade inflammation occurring in the absence of overt infection is defined as "inflammatory aging" and is an important risk factor for morbidity and mortality in the elderly. The nanoparticle compositions of the present application can be used to treat aging. In some embodiments, the method comprises the step of administering to a subject in need of treatment an effective amount of a nanoparticle composition comprising: 0.1-100% (w / v) of an amphiphilic emulsifier; 0-35% (w / v) of a lipophilic or hydrophobic component; a polar liquid carrier; and one or more electrolytes, wherein the nanoparticle composition comprises liposomes and / or micelles having a particle size ranging from 1-800 nm.

[0037] The nanoparticle compositions of the present application can be administered to a subject in need of such treatment via intravenous, intra-arterial, intra-osseous, or intracardiac routes. In certain embodiments, the nanoparticle compositions are administered in an amount of 50-5000 ml, 50-4000 ml, 50-3000 ml, 50-2000 ml, 50-1000 ml, 50-500 ml, 100-5000 ml, 100-4000 ml, 100-3000 ml, 100-2000 ml, 100-1000 ml, 100-500 ml, 200-5000 ml, 200-4000 ml, 200-3000 ml, 200-2000 ml, 200-1000 ml, 200-500 ml, 500-5000 ml, 500-4000 ml, 500-3000 ml, 500-2000 ml, 500-1000 ml, 1000-5000 ml, 1000-4000 ml, 1000-3000 ml, or 1000-2000 ml. In some embodiments, the nanoparticle compositions are administered within 30 seconds to 24 hours in an amount equivalent to about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% of the normal blood volume of the subject. In some embodiments, the nanoparticle compositions of the present application are administered in multiple doses, with each dose separated by 2 hours, 4 hours, 8 hours, 12 hours, 24 hours, or 48 hours.

[0038] In some embodiments, the rate of administration of the nanoparticle composition is 0.1-5000 ml / min, 0.1-2000 ml / min, 0.1-1000 ml / min, 0.1-500 ml / min, 0.1-200 ml / min, 0.1-100 ml / min, 0.1-50 ml / min, 0.1-20 ml / min, 0.1-10 ml / min, 0.1-5 ml / min, 0.1-2 ml / min, 0.1-1 ml / min, 1-5000 ml / min, 1-2000 ml / min, 1-1000 ml / min, 1-500 ml / min, 1-200 ml / min, 1-100 ml / min, 1-50 ml / min, 1-20 ml / min, 1-10 ml / min, 1-5 ml / min, 1-2 ml / min, 2-5000 ml / min, 2-2000 ml / min, 2-1000 ml / min, 2-500 ml / min, 2-200 ml / min, 2-100 ml / min, 2-50 ml / min, 2-20 ml / min, 2-10 ml / min, 2-5 ml / min, 5-5000 ml / min, 5-2000 ml / min, 5-1000 ml / min, 5-500 ml / min, 5-200 ml / min, 5-100 ml / min, 5-50 ml / min, 5-20 ml / min, 5-10 ml / min, 10-5000 ml / min, 10-4000 ml / min, 10-3000 ml / min, 10-2000 ml / min, 10-1000 ml / min, 10-500 ml / min, 10-200 ml / min, 10-100 ml / min, 10-50 ml / min, 20-5000 ml / min, 20-4000 ml / min, 20-3000 ml / min, 20-2000 ml / min, 20-1000 ml / min, 20-500 ml / min, 20-200 ml / min, 20-100 ml / min, 20-50 ml / min, 50-5000 ml / min, 50-4000 ml / min, 50-3000 ml / min, 50-2000 ml / min, 50-1000 ml / min, 50-500 ml / min, 50-200 ml / min, 50-100 ml / min, 100-5000 ml / min, 100-4000 ml / min, 100-3000 ml / min, 100-2000 ml / min, 100-1000 ml / min, 100-500 ml / min, 100-200 ml / min,200-5000 ml / min, 200-4000 ml / min, 200-3000 ml / min, 200-2000 ml / min, 200-1000 ml / min, 200-500 ml / min, 500-5000 ml / min, 500-4000 ml / min, 500-3000 ml / min, 500-2000 ml / min, 500-1000 ml / min, 1000-5000 ml / min, 1000-4000 ml / min, 1000-3000 ml / min, 1000-2000 ml / min, 2000-5000 ml / min, 2000-4000 ml / min, 2000-3000 ml / min, 3000-5000 ml / min, 3000-4000 ml / min, or 4000-5000 ml / min.

[0039] In some embodiments, the nanoparticle composition is administered in an un-oxygenated form. In other embodiments, the nanoparticle composition is an oxygenated nanoparticle composition. In some embodiments, the nanoparticle composition is an oxygenated nanoparticle composition having an oxygen content of 2-50, 2-40, 2-30, 2-20, 2-10, 2-5, 5-50, 5-40, 5-30, 5-20, 5-10, 10-50, 10-40, 10-30, 10-20, 15-50, 15-40, 15-30, 15-20, 20-50, 20-40, 20-30, 25-50, 25-40, 25-30, 30-50, 30-40, or 40-50 ml O2per 100 ml of the nanoparticle composition.

[0040] Other hemodynamic parameters, such as cerebral, renal, cardiac, muscle, spleen, or other tissue perfusion, cardiac output, systolic blood pressure, diastolic blood pressure, mean arterial pressure, stroke volume index, mitochondrial oxidative phosphorylation (monitored by near-infrared spectroscopy or other methods), blood lactate, or membrane polarization, can also be used to determine the "effective amount" of the nanoparticle composition needed to treat a subject for hypercalcitoninemia.

[0041] In some embodiments, the method further comprises administering an additional agent to the subject. The additional agent can be administered prior to, during, or after administration of the nanoparticle of the present application. Examples of additional agents include, but are not limited to, a cardioplegic agent, an inotropic agent, a calcitonin antibody, an osteogepan (a calcitonin gene-related peptide antagonist), sitagliptin (a dipeptidyl peptidase 4 inhibitor used to block calcitonin signaling or activation).

[0042] III. Nanoparticle Compositions

[0043] In one embodiment, the nanoparticle composition for treating or preventing hypercalcitoninemia comprises one or more amphiphilic emulsifiers, a lipophilic or hydrophobic component, a polar liquid carrier, and one or more electrolytes. The amphiphilic emulsifiers form micelles (LMs) that carry the lipophilic or hydrophobic material in a micellar core (LM), which is surrounded by the polar liquid carrier, and / or form liposomes that contain a lipid bilayer with a hydrophilic interior (inside).

[0044] In some embodiments, the nanoparticle composition of the present application comprises micelles and liposomes having a particle size in the range of 1-1000 nm, 1-800 nm, 1-500 nm, 1-400 nm, 1-300 nm, or 1-200 nm as measured by electron microscopy.

[0045] In some embodiments, the nanoparticle composition of the present application comprises: (1) micelles having a particle size in the range of 30-800 nm, 30-500 nm, 30-400 nm, 30-300 nm, 30-200 nm, 30-150 nm, 30-120 nm, 30-100 nm, 30-80 nm, 40-500 nm, 40-400 nm, 40-300 nm, 40-200 nm, 40-150 nm, 40-120 nm, 40-100 nm, 40-80 nm, 50-500 nm, 50-400 nm, 50-300 nm, 50-200 nm, 50-150 nm, 50-120 nm, 50-100 nm, 50-80 nm, 100-500 nm, 100-400 nm, 10-300 nm, 100-200 nm, 100-150 nm, or 100-120 nm as measured by electron microscopy; and (2) liposomes having a particle size in the range of 1-30 nm, 1-25 nm, 1-20 nm, 1-15 nm, 1-10 nm, 3-30 nm, 3-25 nm, 3-20 nm, 3-15 nm, 3-10 nm, 5-30 nm, 5-25 nm, 5-20 nm, 5-15 nm, 5-10 nm, 1-30 nm, 7-25 nm, 7-20 nm, 7-15 nm, 7-10 nm, 10-30 nm, 10-25 nm, 10-20 nm, or 10-15 nm as measured by electron microscopy.

[0046] In some embodiments, the nanoparticle compositions of the present application comprise nanoparticles (including micelles and liposomes) having an average particle size of 1-800 nm, 1-500 nm, 1-300 nm, 1-100 nm, 1-80 nm, 1-50 nm, 1-40 nm, 1-30 nm, 1-25 nm, 1-20 nm, 1-15 nm, 1-10 nm, 1-5 nm, 5-800 nm, 5-500 nm, 5-300 nm, 5-100 nm, 5-80 nm, 5-50 nm, 5-40 nm, 5-30 nm, 5-25 nm, 5-20 nm, 5-15 nm, 5-10 nm, 10-800 nm, 10-500 nm, 10-300 nm, 10-100 nm, 10-80 nm, 10-50 nm, 10-40 nm, 10-30 nm, 10-25 nm, 10-20 nm, 15-800 nm, 15-500 nm, 15-300 nm, 15-100 nm, 15-80 nm, 15-50 nm, 15-40 nm, 15-30 nm, 15-25 nm, 15-20 nm, 20-800 nm, 20-500 nm, 20-300 nm, 20-100 nm, 20-80 nm, 20-50 nm, 20-40 nm, 20-30 nm, 20-25 nm, 25-800 nm, 25-500 nm, 25-300 nm, 25-100 nm, 25-80 nm, 25-50 nm, 25-40 nm, 25-30 nm, 30-800 nm, 30-500 nm, 30-300 nm, 30-100 nm, 30-80 nm, 30-50 nm, 30-40 nm, 40-800 nm, 40-500 nm, 40-300 nm, 40-100 nm, 40-80 nm, 40-50 nm, 50-800 nm, 50-500 nm, 50-300 nm, 50-100 nm, 50-80 nm, 80-800 nm, 80-500 nm, 80-300 nm, or 80-100 nm as measured by electron microscopy. In some embodiments, the nanoparticle compositions of the present application comprise nanoparticles having an average particle size of 16-18 nm, 15-19 nm, or 14-20 nm as measured by electron microscopy.

[0047] In some embodiments, the nanoparticle compositions of the present application comprise nanoparticles (including micelles and liposomes) having an average particle size of 10-1000 nm, 10-800 nm, 10-500 nm, 10-300 nm, 10-200 nm, 10-150 nm, 10-120 nm, 10-100 nm, 10-90 nm, 10-70 nm, 10-50 nm, 10-30 nm, 30-1000 nm, 30-800 nm, 30-500 nm, 30-300 nm, 30-200 nm, 30-150 nm, 30-120 nm, 30-100 nm, 30-90 nm, 30-70 nm, 30-50 nm, 50-1000 nm, 50-800 nm, 50-500 nm, 50-300 nm, 50-200 nm, 50-150 nm, 50-120 nm, 50-100 nm, 50-90 nm, 50-70 nm, 70-1000 nm, 70-800 nm, 70-500 nm, 70-300 nm, 70-200 nm, 70-150 nm, 70-120 nm, 70-100 nm, 70-90 nm, 80-1000 nm, 80-800 nm, 80-500 nm, 80-300 nm, 80-200 nm, 80-150 nm, 80-120 nm, 80-100 nm, 80-90 nm, 90-1000 nm, 90-800 nm, 90-500 nm, 90-300 nm, 90-200 nm, 90-150 nm, 90-120 nm, 90-100 nm, 100-1000 nm, 100-800 nm, 100-500 nm, 100-300 nm, 100-200 nm, 100-150 nm, 100-120 nm, 120-1000 nm, 120-800 nm, 120-500 nm, 120-300 nm, 120-200 nm, 120-150 nm, 150-1000 nm, 150-800 nm, 150-500 nm, 150-300 nm, 150-200 nm, 200-1000 nm, 200-800 nm, 200-500 nm, or 200-300 nm, as measured by dynamic light scattering methods (e.g., using a Malvern Zetasizer model or nano ZS model). In some embodiments, the nanoparticle compositions of the present application comprise nanoparticles having an average particle size of 92-96 nm, 90-98 nm, or 85-105 nm.

[0048] Lipophilic or hydrophobic components are dispersed in a polar liquid carrier to form a nanoemulsion comprising unilamellar micelles (with a polar outer surface, an inner hydrophobic space filled with the lipophilic or hydrophobic components and / or other hydrophobic molecules) and bilayer liposomes (with a polar outer surface and an inner hydrophilic space). Because the solubility of hydrophobic gases such as oxygen and nitric oxide (NO) in the core of the micellar lipid is significantly higher than water or other aqueous environments, the nanoparticle compositions of the present application are capable of delivering oxygen and other hydrophobic gases to tissues of the body.

[0049] The solubility of the hydrophobic gases in the lipophilic or hydrophobic core facilitates their uptake and transport to tissues. Endogenously produced gases (carbon monoxide, nitric oxide, hydrogen sulfide) can also be encapsulated and delivered by the emulsion to modulate vascular tone and apoptosis processes.

[0050] In some embodiments, the nanoparticle composition is an oxygenated nanoparticle composition that enhances aerobic metabolism. In some embodiments, the nanoparticle composition is an oxygenated nanoparticle composition that contains 1-50, 1-40, 1-30, 1-20, 1-10, 1-5, 1-2, 2-50, 2-40, 2-30, 2-20, 2-10, 2-5, 5-50, 5-40, 5-30, 5-20, 5-10, 10-50, 10-40, 10-30, 10-20, 15-50, 15-40, 15-30, 15-20, 20-50, 20-40, 20-30, 25-50, 25-40, 25-30, 30-50, 30-40, or 40-50 ml O2 per 100 ml of the nanoparticle composition.

[0051] In some embodiments, the nanoparticle composition contains nitric oxide (NO) in an amount of 1-50, 1-40, 1-30, 1-20, 1-10, 1-5, 1-2, 2-50, 2-40, 2-30, 2-20, 2-10, 2-5, 5-50, 5-40, 5-30, 5-20, 5-10, 10-50, 10-40, 10-30, 10-20, 15-50, 15-40, 15-30, 15-20, 20-50, 20-40, 20-30, 25-50, 25-40, 25-30, 30-50, 30-40, or 40-50 ml NO per 100 ml of the nanoparticle composition.

[0052] Xenon and argon are hydrophobic gases that can protect the brain in pathological conditions such as epilepsy. In some embodiments, the nanoparticle composition comprises xenon or argon or a combination thereof in an amount of 1-50, 1-40, 1-30, 1-20, 1-10, 1-5, 1-2, 2-50, 2-40, 2-30, 2-20, 2-10, 2-5, 5-50, 5-40, 5-30, 5-20, 5-10, 10-50, 10-40, 10-30, 10-20, 15-50, 15-40, 15-30, 15-20, 20-50, 20-40, 20-30, 25-50, 25-40, 25-30, 30-50, 30-40, or 40-50 ml O2per 100 ml of the nanoparticle composition.

[0053] In some embodiments, the nanoparticle composition of the application further comprises an apoptosis inhibitor (e.g., Z-VAD-FMK, an apoptosis inhibiting peptide), a mitochondrial integrity protecting agent (e.g., cyclosporin A, an inhibitor of mitochondrial inner pore opening), a signal transduction modulator (e.g., diacylglycerol or cyclic guanosine monophosphate), or an antioxidant (e.g., coenzyme Q10).

[0054] When nanoparticles comprising liposomes with an average particle size of less than 30 nm are used, the liposomes can cross endothelial cell layers to enter the interstitial space of tissues. Such liposomes are useful in situations where the permeability of the vascular lumen is not altered or has been increased; where cellular uptake of lipophilic or hydrophobic agents needs to be enhanced; or where internalization of molecules or cellular components that can positively modulate intracellular mechanisms needs to be facilitated.

[0055] In some cases, the nanoparticle composition of the application is capable of exerting osmotic pressure and / or adsorbing tissue-damaging mediators (e.g., prostaglandins, nitric oxide, leukotrienes, thromboxanes) and other lipophilic or hydrophobic mediators (e.g., platelet-activating factor). Thus, in some cases, the nanoparticles of the application are capable of adsorbing toxic molecules produced by patients with hypercalcitoninemia. For example, lymphokines produced by intestinal and thoracic duct lymph nodes can cause acute lung injury and decreased deformability of red blood cells. Other toxic molecules include, but are not limited to, leukotrienes, prostaglandins, nitric oxide, endotoxins, and tumor necrosis factor (TNF). The nanoparticles in the nanoparticle composition are effective at adsorbing lipophilic or hydrophobic chemical mediators. In some cases, this adsorption results from electrostatic interactions. In other cases, the nanoparticles can carry antagonists of the toxic chemical mediators (e.g., antibodies to endotoxins).

[0056] For hyperprocalcitoninemic patients with increased vascular wall permeability due to capillary leakage, the small size of the above phospholipid nanoparticles (PN) is advantageous for their entry into the interstitial space of tissues blocked by larger structures. Mechanisms of capillary leakage include endothelial cell death and neutrophil activation; mediation by cytokines such as IL-1, TNF, and nitric oxide. Neutrophils destroy the cell wall by adhering to the damaged endothelial cells and releasing active oxygen and myeloperoxidase enzymes. Nanoparticles can leak into the interstitial space through capillary leakage and exert anti-inflammatory effects in this interstitial space.

[0057] Preferably, the nanoparticle composition is formulated to contain the lipid-loaded micelles and / or liposomes stable for at least 3 days, 7 days, 2 weeks, 4 weeks, 12 weeks, 20 weeks, 180 days, 30 weeks, 40 weeks, one year, or more at room temperature (e.g., 25°C) or 5°C. Stability can be determined by measuring changes in particle size. Micelles in unstable emulsions will coalesce to form micelles of larger particle size. In certain preferred embodiments, the nanoparticle composition is stable for at least 4 weeks at room temperature.

[0058] In some embodiments, the nanoparticle composition is formed from 5-40% (w / v) soybean oil and 0.1-18% (w / v) egg phospholipid. In some embodiments, the nanoparticle composition further comprises NaCl at a final concentration of 50-200 mM. In some embodiments, the nanoparticle composition further comprises 1-5% glycerol. In one embodiment, the nanoparticle composition comprises 10% (w / v) soybean oil, 0.6% (w / v) egg yolk phospholipid, 1.13% (w / v) glycerol, and 77 mM NaCl. In another embodiment, the nanoparticle composition comprises 20% (w / v) soybean oil, 1.2% (w / v) egg yolk phospholipid, 2.25% (w / v) egg yolk phospholipid. In another embodiment, the nanoparticle composition comprises 20% (w / v) soybean oil, 1.2% (w / v) egg yolk phospholipid, 2.25% (w / v) egg yolk phospholipid, and 77 mM NaCl.

[0059] In some embodiments, the composition consists of: 10-40% (w / v), preferably 15-35% soybean oil; 1-18% (w / v), preferably 10-15% egg phospholipid; electrolytes (sodium chloride and sodium lactate), wherein the electrolyte components total 50 mM to 200 mM; 0.1 mM to 10 mM histidine; and water, such that the egg phospholipid simultaneously forms: (1) lipid-loaded micelles having a lipophilic or hydrophobic inner core in aqueous solution with an average particle size of 1-150 nm, preferably 40 nm to 120 nm, as determined by dynamic light scattering, and stable for at least 4 weeks at room temperature; and (2) liposomes with a particle size range of 1-25 nm as determined by electron microscopy.

[0060] In another embodiment, the nanoparticle composition comprises 10-40% (w / v) soybean oil and 6-18% (w / v) egg yolk phosphatidylcholine or soybean phosphatidylcholine. In some embodiments, the nanoparticle composition further comprises 0.6% (w / v) NaCl, 0.385% (w / v) sodium lactate (L), and 0.155% (w / v) histidine. In some embodiments, the nanoparticle composition comprises 20-30% (w / v) soybean oil, 12% (w / v) egg yolk phosphatidylcholine or soybean phosphatidylcholine, 0.6% (w / v) NaCl, 0.385% (w / v) sodium lactate (L), and 0.155% (w / v) histidine.

[0061] In another embodiment, the nanoparticle composition comprises 20% (w / v) soybean oil, 12% (w / v) egg yolk phosphatidylcholine or soybean phosphatidylcholine, 0.6% (w / v) NaCl, 0.385% (w / v) sodium lactate (L), and 0.155% (w / v) histidine, wherein the nanoparticle composition is prepared under conditions to form nanoparticles (including liposomes and micelles) having an average particle size of 250-300 nm as measured by dynamic light scattering. In some embodiments, the nanoparticles comprise liposomes having a particle size range of 1-25 nm or 7-20 nm as measured by electron microscopy, and micelles having a particle size range of 30-130 nm or 40-100 nm as measured by electron microscopy.

[0062] In another embodiment, the nanoparticle composition comprises 30% (w / v) soybean oil, 12% (w / v) egg yolk phosphatidylcholine or soybean phosphatidylcholine, 0.6% (w / v) NaCl, 0.385% (w / v) sodium lactate (L), and 0.155% (w / v) histidine, wherein the nanoparticle composition is prepared under conditions to form nanoparticles (including liposomes and micelles) having an average particle size of 80-120 nm as measured by dynamic light scattering. In some embodiments, the nanoparticles comprise liposomes having a particle size range of 1-25 nm or 7-20 nm as measured by electron microscopy, and micelles having a particle size range of 30-130 nm or 40-100 nm as measured by electron microscopy.

[0063] Other oil sources can also be used, such as chia seed oil, pumpkin seed oil, and the like. In certain embodiments, the nanoparticle compositions described above can further comprise about 2-40% (w / v), about 2-20% (w / v), about 4-10% (w / v), or about 5% (w / v) of an albumin, an albumin polymer, or an albumin polymer conjugated to an amino acid or a peptide, which is added to the nanoparticle composition after micelle formation. In other embodiments, the hydrophobic or hydrophilic components are entrapped within the red blood cell ghosts.

[0064] In certain embodiments, the lipocalyx comprises 10-40% (w / w) of the nanoparticle composition, and the liposome comprises 5-30% (w / w) of the nanoparticle composition. In some embodiments, the lipocalyx is prepared using soybean oil, and the liposome is prepared using chia seed oil, which has a stronger anti-inflammatory effect than soybean oil.

[0065] In certain embodiments, the nanoparticle composition of the present application comprises: a lipophilic or hydrophobic component selected from the group consisting of soybean oil, chia seed oil, and algal oil; an emulsifier selected from the group consisting of phospholipids and alpha-phosphatidylcholine; and an amino acid or n-acetylamino acid at a final concentration of 0.2-20 mM, 0.5-10 mM, 0.5-5 mM, or 0.5-2 mM.

[0066] In certain embodiments, the nanoparticle composition has a final amino acid concentration of 0.000001-10 mM, 0.01-10 mM, 0.1-10 mM, 0.2-10 mM, 0.5-10 mM, 1-10 mM, 2.5-10 mM, 5-10 mM, or 7.5-10 mM. In certain embodiments, the nanoparticle composition has a final amino acid concentration of 0.001, 0.01, 0.1, 0.2, 0.5, 1, 2.5, 5, 7.5, or 10 mM. The ratio of emulsifier to lipophilic or hydrophobic component (w / w) can range from about 1:400 to about 1:1, preferably from about 1:200 to about 1:50. In one embodiment, the ratio of emulsifier to lipophilic or hydrophobic component (w / w) is about 1:100. In another embodiment, the ratio of emulsifier to lipophilic or hydrophobic component (w / w) is about 1.2:100.

[0067] In some embodiments, the nanoparticle composition consists essentially of liposomes, without a lipophilic or hydrophobic component such as soybean oil.

[0068] In some embodiments, the nanoparticle composition encapsulates one or more active pharmaceutical ingredients or agents (e.g., nucleic acids, proteins, small molecule drugs, etc.) in the lipocalyx and / or the liposome. The active pharmaceutical ingredients or agents can be incorporated into the lipophilic or hydrophobic core of the lipocalyx or liposome, or into the hydrophilic core of the liposome.

[0069] In one embodiment, the nanoparticle composition comprises soybean oil, egg yolk phosphatidylcholine, an amino acid, beta-endorphin, or other modulator (at a final concentration of 0.1 fM to 10 mM, with an effect at a femtomolar concentration).

[0070] The nanoparticle compositions of the present application do not contain hemoglobin, derivatives thereof, perfluorocarbons, and derivatives thereof. As used herein, "the compositions do not contain hemoglobin, derivatives thereof, perfluorocarbons, and derivatives thereof" means that the compositions do not contain any hemoglobin, derivatives thereof, perfluorocarbons, and derivatives thereof, or that the compositions contain less than 0.1% w / v of hemoglobin, derivatives thereof, perfluorocarbons, and derivatives thereof.

[0071] The nanoparticle compositions of the present application generally do not contain Ca ++ , K + , Mg ++ , and added Al +++ . In certain embodiments, Ca ++ and K + are added to the nanoparticle compositions prior to use (e.g., within 24 hours prior to use). In other embodiments, Ca ++ is premixed with the nanoparticle compositions. Because Al +++ is toxic to bone marrow, brain, hematopoiesis, heme synthesis, globin synthesis, iron absorption and metabolism, and fetal development, all oils and other components should be minimized for Al +++ content. In certain embodiments, the concentration of Al +++ in the nanoparticle compositions is less than 25 mg / 1, 20 mg / 1, 10 mg / 1, or 5 mg / 1. In other embodiments, the nanoparticle compositions are completely free of Al +++ , i.e., undetectable by conventional methods.

[0072] In certain embodiments, the micelles in the nanoparticle compositions of the present application are free micelles, not encapsulated by any type of particle. Further, the micelles have a wall composed of a single or double layer of amphiphilic emulsifier molecules, such that the micelles can easily fuse with the cell membranes of tissues that contact the nanoparticle compositions. Further, the micelles in the nanoparticle compositions of the present application do not contain hemoglobin, derivatives thereof, perfluorocarbons, and derivatives thereof.

[0073] Amphiphilic emulsifier

[0074] The amphiphilic emulsifier can be any amphiphile or amphiphilic molecule, having a hydrophobic tail located in the lipophilic or hydrophobic core of the micelle, and a hydrophilic end in contact with the polar carrier.

[0075] As used herein, the term "amphiphile" refers to a compound that has both hydrophilic and lipophilic or hydrophobic properties. Examples of amphiphiles include, but are not limited to, natural amphiphiles such as phospholipids, cholesterol, glycolipids, fatty acids, bile acids, saponins, and synthetic amphiphiles such as amphiphilic peptides.

[0076] Examples of phospholipids include: natural / synthetic phospholipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidic acid, phosphatidylglycerol, phosphatidylinositol, lysophosphatidylcholine, sphingomyelin, egg lecithin, soy lecithin, hydrogenated phospholipids.

[0077] Examples of glycolipids include: glyceroglycolipids, glycosphingolipids. Examples of glyceroglycolipids include: digalactosyl diglyceride (e.g., digalactosyl dilauryl glyceride, digalactosyl dimyristyl glyceride, digalactosyl dipalmitoyl glyceride, digalactosyl distearoyl glyceride), galactosyl diglyceride (e.g., galactosyl dilauryl glyceride, galactosyl dimyristyl glyceride, galactosyl dipalmitoyl glyceride, galactosyl distearoyl glyceride). Examples of glycosphingolipids include: galactocerebroside, lactosylceramide, ganglioside.

[0078] Examples of sterols include: cholesterol, cholesterol hemisuccinate, 3β-[N-(N',N'- dimethylaminoethane)carbamoyl]cholesterol, ergosterol, lanosterol.

[0079] In one embodiment, the emulsifier comprises lecithin, or in other words, egg lecithin. In another embodiment, the emulsifier is soy lecithin or alpha-phosphatidylcholine.

[0080] In other embodiments, the emulsifier can comprise 0.1-100%, 0.1-90%, 0.1-80%, 0.1-70%, 0.1-60%, 0.1-50%, 0.1-40%, 0.1-30%, 0.1-20%, 0.1-15%, 0.1-10%, 0.1-5%, 0.1-2%, 0.3-100%, 0.3-90%, 0.3-80%, 0.3-70%, 0.3-60%, 0.3-50%, 0.3-40%, 0.3-30%, 0.3-20%, 0.3-15%, 0.3-10%, 0.3-5%, 0.3-2%, 0.6-100%, 0.6-90%, 0.6-80%, 0.6-70%, 0.6-60%, 0.6-50%, 0.6-40%, 0.6-30%, 0.6-20%, 0.6-15%, 0.6-10%, 0.6-5%, 0.6-2%, 2-100%, 2-90%, 2-80%, 2-70%, 2-60%, 2-50%, 2-40%, 2-30%, 2-20%, 2-15%, 2-10%, 2-5%, 6-100%, 6-90%, 6-80%, 6-70%, 6-60%, 6-50%, 6-40%, 6-30%, 6-20%, 6-15%, 6-10%, 10-100%, 10-90%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 10-15%, 15-100%, 15-90%, 15-80%, 15-70%, 15-60%, 15-50%, 15-40%, 15-30%, 15-20%, 20-100%, 20-90%, 20-80%, 20-70%, 20-60%, 20-50%, 20-40%, 20-30%, 30-100%, 30-90%, 30-80%, 30-70%, 30-60%, 30-50%, 30-40%, 40-100%, 40-90%, 40-80%, 40-70%, 40-60%, 40-50%, 50-100%, 50-90%, 50-80%, 50-70%, 50-60% (w / v or v / v) of the nanoparticle composition. In some embodiments, the emulsifier can comprise more than 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% (w / v or v / v) of the nanoparticle composition.In certain embodiments, the emulsifier is present in the nanoparticle composition in an amount of about 1%, 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 18%, about 20% (w / v or v / v), or a range between any two of the foregoing. In other embodiments, the emulsifier is present in the nanoparticle composition in an amount of about 7-9%, 9-11%, 11-13%, 13-15%, 15-17%, 17-19%, 10-14%, 9-15%, or 8-16% (w / v or v / v), or a range between any two of the foregoing. In still other embodiments, the upper and / or lower limit is defined by any of the concentrations listed herein.

[0081] In certain preferred embodiments, the emulsifier is lecithin (e.g., egg yolk lecithin or soy lecithin) in an amount of any of the foregoing values or ranges.

[0082] Lipophilic or hydrophobic component

[0083] The term "lipophilic component" as used herein refers to a naturally occurring or non-naturally occurring lipid-soluble substance. Examples of lipophilic components include, but are not limited to, fatty acyl groups, glycerides, phospholipids, sphingolipids, sterol lipids, prenol lipids, glycolipids, polyketides, non-natural lipids, cationic lipids, amphiphilic alkyl amino acid derivatives, dialkyldimethylammonium, polyglycerol alkyl ethers, polyoxyethylene alkyl ethers, tri-n-octylamine, boric acid, tris(3,5-dimethyl-4-heptyl) borate, triglycerides, diglycerides, and other acylglycerols (e.g., tetraglycerol, pentaglycerol, hexaglycerol, heptaglycerol, octaglycerol, nonaglycerol, and decaglycerol), hydrophobic peptides, hydrophobic polysaccharides, silicones, lipopeptides, cyclic peptides, and mixtures thereof. In certain embodiments, the lipophilic or hydrophobic component comprises soybean oil, chia seed oil, or algal oil.

[0084] In one embodiment, the hydrophobic component is soybean oil. The hydrophobic component can also be derived from chia seeds, which are rich in anti-inflammatory omega-3 fatty acids. Soybean oil is prothrombotic and procoagulant, and is therefore preferred when coagulation is desired. When the risk of bleeding has subsided, omega-3 fatty acid-rich oils are preferred because of their anti-thrombotic properties. Omega-3 fatty acid-rich oils include, but are not limited to, chia seed oil, algal oil, pumpkin seed oil, flaxseed oil, or fish oil.

[0085] In certain embodiments, the lipophilic or hydrophobic component comprises an unsaturated fatty acid having one or more cis or trans configured alkenyl functional groups. The cis configuration refers to adjacent hydrogen atoms or other groups being on the same side of the double bond. In the trans configuration, the groups are on opposite sides of the double bond. The rigidity of the double bond fixes its configuration, resulting in a bending of the molecular chain and limiting the conformational freedom of the fatty acid in the case of cis isomers. Generally, the more double bonds a molecular chain contains, the less flexible it is. When a molecular chain contains multiple cis bonds, its most stable conformation is significantly bent. For example, oleic acid, which contains one double bond, forms a "kink," and linoleic acid, which contains two double bonds, is bent more dramatically. Alpha-linolenic acid, which contains three double bonds, tends to be hook-shaped. As a result, in a constrained environment, such as when the fatty acid is a component of a phospholipid in a lipid bilayer or a triglyceride in a lipid droplet, cis bonds limit the ability of the fatty acid to pack tightly, thereby affecting the structure of the membrane or the melting point of the fat. In some embodiments, the lipophilic or hydrophobic component comprises up to 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% (w / v or v / v) of an unsaturated fatty acid having one or more cis configured alkenyl functional groups.

[0086] Examples of cis unsaturated fatty acids include, but are not limited to, 4-decenoic acid, 5-dodecenoic acid, brassidic acid, palmitoleic acid, oleic acid, elaidic acid, vaccenic acid, petroselinic acid, gadoleic acid, eicosenoic acid, erucic acid, cetoleic acid, nervonic acid, ximenic acid, triacontenoic acid; n-3 type unsaturated fatty acids such as alpha-linolenic acid, stearidonic acid, eicosatetraenoic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid; n-6 type unsaturated fatty acids such as linoleic acid, elaidic acid, gamma-linolenic acid, dihomo-gamma-linolenic acid, arachidonic acid; conjugated fatty acids such as conjugated linoleic acid, alpha-eleostearic acid; fatty acids containing a double bond in the 5 position such as pinolenic acid, abietic acid, juniperonic acid, parinaric acid; other polyvalent unsaturated fatty acids such as hexadecatrienoic acid, linolenelaidic acid, clupadonic acid, nicholic acid; branched-chain fatty acids such as isobutyric acid, isovaleric acid, isoformic acid, trans-isoformic acid; hydroxy fatty acids such as alpha-hydroxy acid, beta-hydroxy acid, mycolic acid, polyhydroxy acid; epoxy fatty acids; keto fatty acids; and cyclic fatty acids. In certain embodiments, the lipophilic or hydrophobic component further comprises an amphiphilic molecule.

[0087] The proportion of the lipophilic or hydrophobic component to the nanoparticle composition can be about 0-80%, 1-80%, 1-70%, 1-60%, 1-50%, 1-40%, 1-30%, 1-20%, 5-80%, 5-70%, 5-60%, 5-50%, 5-40%, 5-30%, 5-20%, 10-80%, 10-70%, 10-60%, 10-50%, 10-40%, 10-30%, 10-20%, 15-80%, 15-70%, 15-60%, 15-50%, 15-40%, 15-30%, 15-20%, 20-80%, 20-70%, 20-60%, 20-50%, 20-40%, 20-30%, 30-80%, 30-70%, 30-60%, 30-50%, 30-40%, 40-80%, 40-70%, 40-60%, 40-50%, 40-80%, 40-70%, 40-60%, 40-30%, 50-80%, 50-70%, 50-60%, 60-80%, 60-70%, or 70-80% (w / v or v / v). In certain embodiments, the proportion of the lipophilic or hydrophobic component to the nanoparticle composition is about 10%, about 15%, about 20%, about 25%, about 30%, and about 35% (w / v or v / v). In some embodiments, the proportion of the lipophilic or hydrophobic component to the nanoparticle composition is 0-35%, 5-35%, 10-35%, 15-35%, 20-35%, 25-35%, 30-35%, 0-30%, 5-30%, 10-30%, 15-30%, 20-30%, 25-30%, 0-25%, 5-25%, 10-25%, 15-25%, 20-25%, 0-15%, 5-15%, or 10-15% (w / v or v / v), or a percentage range combination comprising any integer value selected from the group consisting of 10%, 15%, 20%, 25%, 30%, or 35% (w / v or v / v). In other embodiments, the upper and / or lower limit of the lipophilic or hydrophobic component is defined by any of the concentrations listed herein.

[0088] Polar liquid carrier

[0089] The polar liquid carrier can be any pharmaceutically acceptable polar liquid capable of forming an emulsion with the lipids. The term "pharmaceutically acceptable" refers to molecular entities and compositions that are of sufficient purity and quality for use in formulating compositions or medicaments of the present application, and which do not produce an adverse, allergic or other untoward reaction when appropriately administered to an animal or a human. Since both human (clinical and non-prescription) and veterinary uses are included within the scope of the present application, the pharmaceutically acceptable formulations should include compositions or medicaments for use on humans or animals. In one embodiment, the polar liquid carrier is water or a water-based solution. In another embodiment, the polar liquid carrier is a non-aqueous polar liquid (such as dimethyl sulfoxide, polyethylene glycol, and polar silicone liquids).

[0090] Water-based solutions generally comprise a physiologically compatible electrolyte carrier that is isotonic or near isotonic with whole blood. The carrier can be, for example, normal saline, a saline-glucose mixture, Ringer's solution, Ringer's lactate solution, Locke-Ringer's solution, Krebs-Ringer's solution, Hartmann's balanced salt solution, heparinized sodium citrate-citric acid-dextrose solution, and polymeric plasma substitutes (such as polyethylene oxide, polyvinylpyrrolidone, polyvinyl alcohol, ethylene oxide-propylene glycol condensates). The nanoparticle compositions can also comprise other components (such as pharmaceutically acceptable carriers, diluents, fillers, and salts), the selection of which depends on the dosage form used, the condition being treated, the particular purpose to be achieved according to the judgment of the ordinarily skilled person, and the nature of such additives.

[0091] Electrolytes

[0092] In one embodiment, the nanoparticle compositions of the present application comprise one or more electrolytes. Electrolytes used in the present application generally include various medical electrolytes. Examples of electrolytes include: sodium salts (such as sodium chloride, sodium bicarbonate, sodium citrate, sodium lactate, sodium sulfate, sodium dihydrogen phosphate, sodium phosphate dibasic, sodium acetate, sodium glycerophosphate, sodium carbonate, sodium salts of amino acids, sodium propionate, sodium hydroxybutyrate, sodium gluconate); potassium salts (such as potassium chloride, potassium acetate, potassium gluconate, potassium bicarbonate, potassium glycerophosphate, potassium sulfate, potassium lactate, potassium iodide, potassium dihydrogen phosphate, potassium phosphate dibasic, potassium citrate, potassium salts of amino acids, potassium propionate, potassium hydroxybutyrate); calcium salts (such as calcium chloride, calcium gluconate, calcium lactate, calcium glycerophosphate, calcium pantothenate, calcium acetate); magnesium salts (such as magnesium chloride, magnesium sulfate, magnesium glycerophosphate, magnesium acetate, magnesium lactate, magnesium salts of amino acids); ammonium salts (such as ammonium chloride); zinc salts (such as zinc sulfate, zinc chloride, zinc gluconate, zinc lactate, zinc acetate); iron salts (such as ferric sulfate, ferric chloride, and ferrous gluconate); copper salts (such as copper sulfate); and manganese salts (such as manganese sulfate). Among these, particularly preferred are sodium chloride, potassium chloride, magnesium chloride, sodium phosphate dibasic, potassium phosphate dibasic, potassium dihydrogen phosphate, sodium lactate, sodium acetate, sodium citrate, potassium acetate, potassium glycerophosphate, calcium gluconate, calcium chloride, magnesium sulfate, zinc sulfate.

[0093] The concentration of calcium, sodium, magnesium or potassium ions is generally in the range of their normal physiological concentrations in blood plasma. Typically, the target concentrations of these ions are obtained by dissolving calcium chloride, sodium chloride and magnesium chloride. Sodium ions can also be derived from dissolved organic sodium salts in the solution.

[0094] In one embodiment, the electrolyte comprises sodium chloride, sodium lactate, or both.

[0095] In particular embodiments, the percentage concentration of sodium chloride in the nanoparticle composition is about 0.2-1%, 0.3-0.9%, 0.4-0.8%, 0.5-0.7%, or about 0.6% (w / v).

[0096] In another embodiment, the concentration of sodium chloride in the nanoparticle composition is 50-150 mM, 70-130 mM, 80-120 mM, 90-110 mM, 95-100 mM, or about 97.4 mM.

[0097] In another embodiment, the percentage concentration of sodium L-lactate, sodium D-lactate, or a mixture thereof in the nanoparticle composition is about 0.1-0.7%, 0.2-0.6%, 0.3-0.5%, 0.35-0.45%, 0.38-0.39%, or about 0.385% (w / v).

[0098] In another embodiment, the concentration of sodium L-lactate, sodium D-lactate, or a mixture thereof in the nanoparticle composition is 10-60 mM, 20-50 mM, 30-40 mM, or about 34 mM.

[0099] In one embodiment, the sodium ion concentration ranges from about 70-180 mM, 90-170 mM, 70-160 mM, 100-160 mM, 110-150 mM, 120-140 mM, 125-135 mM, 131-133 mM, or about 131.4 mM.

[0100] In one embodiment, the calcium ion concentration ranges from about 0.5-4.0 mM, 0.5-1.0 mM, 0.5-2 mM, 0.5-3 mM, 1-2 mM, 1-3 mM, 1-4 mM, 2-2.5 mM, 2-3 mM, 2-4 mM, 2.5-3 mM, or 3-4 mM.

[0101] In one embodiment, the magnesium ion concentration ranges from 0 to 10 mM. In another embodiment, the magnesium ion concentration ranges from about 0.3-0.45 mM, 0.3-0.35 mM, 0.3-0.4 mM, 0.35-0.4 mM, 0.35-0.4 mM, 0.35-0.4 mM, or 0.4-0.45 mM. It is undesirable to include excess magnesium ions in the nanoparticle composition of the present application because high magnesium ion concentrations can impair the strength of cardiac contractions. In a preferred embodiment of the present application, the nanoparticle composition comprises sub-physiological levels of magnesium ions.

[0102] In one embodiment, the potassium ion concentration ranges from 0-5 mEq / 1 K+ (0-5 mM), preferably a sub-physiological level of 2-3 mEq / 1 K+ (2-3 mM). Thus, the nanoparticle composition is capable of diluting the potassium ion concentration in stored transfusion blood, thereby making it easier to control high potassium ion concentrations and the cardiac arrhythmias and cardiac insufficiency that they can induce. The nanoparticle composition containing sub-physiological amounts of potassium can also be used for blood replacement and for the purpose of hypothermic maintenance of a subject.

[0103] In one embodiment, the chloride ion concentration ranges from 50-200 mM, 50-150 mM, 70-180 mM, 70-130 mM, 80-170 mM, 80-120 mM, 90-160 mM, 90-110 mM, 95-150 mM, 95-100 mM, or about 97.4 mM. In another embodiment, the chloride ion concentration ranges from 110 mM to 125 mM.

[0104] Other ion sources include: sodium salts (e.g., sodium bicarbonate, sodium citrate, sodium lactate, sodium sulfate, sodium dihydrogen phosphate, sodium hydrogen phosphate, sodium acetate, sodium glycerophosphate, sodium carbonate, sodium salts of amino acids, sodium propionate, sodium 3-hydroxybutyrate, sodium gluconate); potassium salts (e.g., potassium acetate, potassium gluconate, potassium bicarbonate, potassium glycerophosphate, potassium sulfate, potassium lactate, potassium iodide, potassium dihydrogen phosphate, potassium hydrogen phosphate, potassium citrate, potassium salts of amino acids, potassium propionate, potassium 3-hydroxybutyrate); calcium salts (e.g., calcium gluconate, calcium lactate, calcium glycerophosphate, calcium pantothenate, calcium acetate); magnesium salts (e.g., magnesium sulfate, magnesium glycerophosphate, magnesium acetate, magnesium lactate, magnesium salts of amino acids); ammonium salts; zinc salts (e.g., zinc sulfate, zinc chloride, zinc gluconate, zinc lactate, zinc acetate); iron salts (e.g., iron sulfate, iron chloride, ferrous gluconate); copper salts (e.g., copper sulfate); and manganese salts (e.g., manganese sulfate). Among these, sodium chloride, potassium chloride, magnesium chloride, sodium hydrogen phosphate, potassium hydrogen phosphate, potassium dihydrogen phosphate, sodium lactate, sodium acetate, sodium citrate, potassium acetate, potassium glycerophosphate, calcium gluconate, calcium chloride, magnesium sulfate, choline chloride, zinc sulfate are particularly preferred.

[0105] Gas carrying capacity of the nanoparticle composition

[0106] The lipophilic or hydrophobic component (present as micelles and / or ghost forms) in the nanoparticle composition enables it to carry more lipophilic gas than a pure aqueous solution. In particular, the lipophilic gas is dissolved in the lipophilic portion of the nanoparticle composition, forming a homogeneous solution with the lipophilic or hydrophobic component and other hydrophobic liquid substances that can be present in this portion.

[0107] In one embodiment, the lipophilic gas is oxygen. The solubility of oxygen in lipids is 4.41 times that in water (Battino et al., J. Am. Oil Chem. Soc. 45:830-833 (1968)). Thus, a nanoparticle composition with a higher lipid content can carry more oxygen than a nanoparticle composition with a lower lipid content. In one embodiment, the lipid content of the nanoparticle composition is about 1-80% (w / v). In other embodiments, the lipid content of the nanoparticle composition is about 10-80% (w / v), 20-60% (w / v), about 20-50% (w / v), about 20-40% (w / v), or about 20-25% (w / v). In another embodiment, the lipid content of the nanoparticle composition is about 21.8%. In certain embodiments, the nanoparticle composition is prepared by mixing the lipophilic or hydrophobic component with the polar liquid component in air at ambient pressure. In other embodiments, additional oxygenation of the nanoparticle composition is performed by bubbling ambient air or pure oxygen into the nanoparticle composition for a predetermined time. Because the presence of gas bubbles in the circulatory system can lead to air embolism, a bubble trap must be added to remove the gas bubbles, leaving only the gas dissolved in the micelle core, the polar carrier, or bound to the protein or other additives. The gas can also be loaded into the micelles by equilibrating the micelles with a gas-rich atmosphere while gently agitating the nanoparticle composition in a mixing chamber to avoid bubble generation. The loading process can also be performed at a pressure greater than 1 atm, followed by a pressure release to release excess gas.

[0108] In another embodiment, the lipophilic gas is xenon (Xe) or argon (Ar). In another embodiment, the lipophilic gas is nitric oxide (NO). In another embodiment, the lipophilic gas is hydrogen sulfide (H2S). In another embodiment, the lipophilic gas is carbon monoxide (CO).

[0109] In one embodiment, the nanoparticle composition comprises micelles loaded with a mixed gas (e.g., a mixture of oxygen, hydrogen sulfide, carbon monoxide, and / or nitric oxide). In another embodiment, the nanoparticle composition comprises a mixture of micelles loaded with different gases. For example, the mixture of micelles can comprise 50% micelles loaded with NO and 50% micelles loaded with O2.

[0110] Rigid non-planar molecules

[0111] The nanoparticle composition can further comprise a molecule having a rigid non-planar structure. Such molecules will increase steric hindrance in the hydrophobic inner core of the micelle structure and increase the gas molecule accommodation space, thereby altering the gas carrying capacity of the micelle. Examples of such molecules include, but are not limited to: (+) naloxone, (+) morphine, (+) naltrexone.

[0112] In one embodiment, the rigid non-planar structure molecule is (+) naloxone, which, unlike the opioid receptor antagonist (-) naloxone, does not bind to opioid receptors and does not exacerbate pain as (-) naloxone does. In another embodiment, the (+) naloxone is used at a concentration of 10 -5 -10 -4 M. In another embodiment, the (+) naloxone is used at a concentration of 10 -4 or more.

[0113] In one embodiment, the (+) naloxone exerts an anti-inflammatory effect at a concentration of 10 -5 -10 -4 M.

[0114] Molecules having a non-planar structure also include branched organic molecules. Examples of such molecules include, but are not limited to: tri-n-octylamine, tri-n-hexylamine, boric acid, tris(3,5-dimethyl-4-heptyl)borate, metal complexed / non-metal complexed meso-porphyrin dimethyl ester and its derivatives, hexaphenyl thiophene, silicone polymers.

[0115] Plasma component

[0116] The nanoparticle composition can further comprise a plasma component. In one embodiment, the plasma is animal plasma. In another embodiment, the plasma is human plasma. While not limited to a particular scientific theory, intravascular infusion of fluids can dilute the concentration of clotting factors to an undesirable level. Thus, the use of plasma as a diluent for the oxygen carrying component can avoid this problem. The plasma can be collected by any method known in the art, but must be substantially free of red blood cells, white blood cells, and platelets. Preferably, an automated plasma separation device is used to obtain the plasma. Plasma separation devices are commercially available, including, for example, devices that separate plasma from blood by ultrafiltration or centrifugation. Plasma separation devices based on ultrafiltration, such as the Auto C A200 (manufacturer: Baxter International Inc., Deerfield, IL, USA), are suitable because they effectively remove red blood cells, white blood cells, and platelets while preserving clotting factors.

[0117] The plasma can be collected in conjunction with an anticoagulant, of which many are known in the art. Preferably, the anticoagulant is one that can chelate calcium, such as citrate. In one embodiment, sodium citrate is used as the anticoagulant at a final concentration of 0.2-0.5%, preferably 0.3-0.4%, and most preferably 0.38%. The plasma can be fresh plasma, frozen plasma, pooled plasma, and / or sterilized plasma. While exogenous plasma is preferred, autologous plasma collected from the subject prior to formulating and administering the nanoparticle composition is also within the scope of the application.

[0118] In addition to naturally derived plasma, synthetic plasma can also be used. The term "synthetic plasma" as used herein refers to an aqueous solution comprising at least one plasma protein. Proteins similar to plasma proteins are also suitable.

[0119] Colloid agent

[0120] In one embodiment, the nanoparticle composition comprises a colloidal osmotic agent in addition to the nanoparticles (micelles or liposomes). The colloidal agent is composed of molecules large enough in molecular weight that they cannot pass through the fenestrations of the capillary bed into the interstitial space of the body tissues, thereby avoiding loss from the circulation. Examples of colloidal agents include, but are not limited to, dextran (e.g., low molecular weight dextran), dextran derivatives (e.g., carboxymethyl dextran, carboxy dextran, cationic dextran, and dextran sulfate), hydroxyethyl starch, hydroxypropyl starch, branched / unsubstituted or substituted starches, gelatin (e.g., modified gelatin), albumin (e.g., human plasma, human serum albumin, heat denatured human plasma protein, and recombinant human serum albumin), PEG, polyvinylpyrrolidone, carboxymethyl cellulose, gum arabic, glucose, dextrose (e.g., dextrose monohydrate), oligosaccharides (e.g., oligosaccharides), polysaccharide degradation products, amino acid and protein degradation products. Among these, low molecular weight dextran, hydroxyethyl starch, modified gelatin, and recombinant albumin are particularly preferred.

[0121] Albumin is desirable because of its antioxidant properties, which minimize the interaction of reactive oxygen species with the micelle components and stabilize the micelle structure. In one embodiment, the colloidal agent is about 2%, 5%, 7%, or 10% (w / v) albumin. In another embodiment, the colloidal agent is a polysaccharide (e.g., dextran) having a molecular weight of 30,000 to 50,000 Daltons (D). In another embodiment, the colloidal agent is a polysaccharide (e.g., dextran) having a molecular weight of 50,000 to 70,000 Daltons (D). High molecular weight dextran solutions are more effective at preventing tissue swelling because of their lower capillary permeability.

[0122] In one embodiment, the concentration of the polysaccharide (in combination with sodium chloride, calcium chloride and magnesium chloride, organic ions of organic sodium salts, and the hexose sugars described above) is sufficient to bring the oncotic pressure of the colloid to near normal human serum levels, about 28 mm Hg.

[0123] In another embodiment, the colloidal agent is glycerol or mannitol at a concentration of about 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 10%, 15%, 20%, 25%, or 30% (w / v) of the nanoparticle composition. In other embodiments, the nanoparticle composition comprises 2-5% w / v glycerol or mannitol.

[0124] Crystallizing agents

[0125] The nanoparticle composition can also comprise a crystallizing agent. The crystallizing agent can be any crystalline substance that is preferably capable of bringing the osmotic pressure in the nanoparticle composition to greater than 800 mOsm / l, i.e., to a "hyperosmolar" state of the nanoparticle composition. Examples of suitable crystalline substances and concentrations thereof in the nanoparticle composition include, but are not limited to, 3% w / v NaCl, 7% NaCl, 7.5% NaCl, 7.5% NaCl with 6% w / v dextran. In one embodiment, the osmotic pressure of the nanoparticle composition is between 800 and 2400 mOsm / l.

[0126] Anti-inflammatory agents and immunomodulators

[0127] In one embodiment, the nanoparticle composition of the present application further comprises an anti-inflammatory agent or an immunomodulator. Examples of anti-inflammatory agents that have been shown to inhibit reactive oxygen species include, but are not limited to, histidine, albumin, (+) naloxone, prostaglandin D2, benzylamine molecules. Other anti-inflammatory compounds and immunomodulatory drugs include: interferons; interferon derivatives, such as Betaseron (beta-interferon); prostaglandin derivatives, such as iloprost, cicaprost; glucocorticoids, including hydrocortisone, prednisolone, methylprednisolone, dexamethasone; immunosuppressants, such as cyclosporin A, methotrexate, azathioprine, sulfasalazine, mycophenolate mofetil; lipoxygenase inhibitors, such as Zileuton, MK-886, WY-50295, SC-45662, SC-41661A, BI-L-357; leukotriene antagonists; peptide derivatives, such as adrenocorticotropic hormone (ACTH) and analogs thereof; soluble TNF receptors; anti-TNF antibodies; soluble receptors for interleukins or other cytokines; antibodies against interleukin or other cytokine receptors; T-cell proteins; calcipotriol and analogs thereof. The above substances can be used alone or in combination.

[0128] Carbohydrates and amino acids

[0129] The nanoparticle composition can include a carbohydrate or mixture thereof. Suitable carbohydrates include, but are not limited to, monohexoses (e.g., glucose, fructose, galactose), mannitol, sorbitol, or other carbohydrates known in the art. In one embodiment, the nanoparticle composition includes physiological levels of hexose. By "physiological levels of hexose" is meant a hexose concentration between 2 mM and 50 mM. In one embodiment, the nanoparticle composition includes 5 mM glucose. In certain instances, it can be desirable to increase the concentration of hexose to provide nutritional support to the cells. Thus, the concentration of hexose can be increased to about 50 mM, if necessary, to provide the caloric requirements for basal nutrition.

[0130] Other suitable carbohydrates include various sugars used for medical purposes. Examples of such sugars include xylitol, dextrin, glycerol, sucrose, trehalose, glycerol, maltose, lactose, and erythritol.

[0131] The nanoparticle composition can include one or more amino acids and / or one or more oligopeptides. Suitable amino acids include, but are not limited to, alanine, arginine, aspartic acid, asparagine, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, threonine, tryptophan, valine, and 2-aminopentenoic acid. In one embodiment, the amino acid is selected from the group consisting of histidine, tyrosine, phenylalanine, and cysteine. In another embodiment, the nanoparticle composition includes one or more amino acids known to inhibit apoptosis. Examples of such amino acids include glutamine, glycine, proline, and 2-aminopentenoic acid.

[0132] The amino acid can be used at a concentration of 0.1 fM to 200 mM, 0.1 fM to 100 pM, 100 pM to 10 nM, 10 nM to 10 μM, 0.01 to 200 mM, 0.2 to 50 mM, or 0.5 to 2 mM. In one embodiment, the amino acid is used at a concentration of 1 mM.

[0133] Buffering agents

[0134] The nanoparticle composition of the present application can further include a biological buffer for maintaining the pH of the fluid within the physiological range of pH 7 to 8. Examples of biological buffers include, but are not limited to, N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES), 3-(N-morpholino)propanesulfonic acid (MOPS), 2-([2-hydroxy-1,1-bis(hydroxymethyl)ethyl]amino)ethanesulfonic acid (TES), 3-[N-tris(hydroxymethyl)methylamino]-2-hydroxyethyl]-1-piperazinepropanesulfonic acid (EPPS), tris[hydroxymethyl]aminomethane (THAM), and tris[hydroxymethyl]aminomethane (TRIS).

[0135] In one embodiment, the buffering agent is histidine, imidazole, a substituted histidine that retains the imidazole ring zwitterionic site, or an imidazole compound, an oligopeptide containing histidine or glycine (e.g., diglycine), or a mixture thereof. Histidine also reduces reactive oxygen species and inhibits cell shrinkage. (See, e.g., Simpkins et al., J. Trauma and Acute Care Surg. 63:565-572 (2007)). Histidine or imidazole can be used at a concentration of about 1 mM, 5 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, or a range of about 0.1 mM to about 200 mM, 1 mM to about 100 mM, 5 mM to about 50 mM, 5 mM to about 20 mM, or any other range between the concentrations of histidine listed above.

[0136] In another embodiment, the nanoparticle compositions of the application maintain physiological pH in vivo using conventional biological components. Briefly, certain biological compounds (e.g., lactic acid) are metabolized in vivo and work in conjunction with other biological components to maintain a physiologically appropriate pH in an animal. These biological components are effective in maintaining a physiologically appropriate pH under conditions of hypothermia and hypovolemia. Examples of conventional biological components include, but are not limited to, carboxylic acids, carboxylate salts, and carboxylate esters. Carboxylic acids have the general structure RCOOX, where R is a branched or straight chain alkyl, alkenyl, or aryl group containing 1 to 30 carbon atoms, which can be substituted, X is hydrogen, sodium, or other biocompatible ionic substituent that can be attached to the oxygen atom, or a short straight or branched chain alkyl group containing 1 to 4 carbon atoms, such as -CH3, -CH2CH3. Examples of carboxylic acids and carboxylate salts include, but are not limited to, lactic acid and sodium lactate, citric acid and sodium citrate, gluconic acid and sodium gluconate, pyruvic acid and sodium pyruvate, succinic acid and sodium succinate, and acetic acid and sodium acetate.

[0137] Coagulation Enhancers

[0138] In certain embodiments, the nanoparticle compositions can further comprise one or more coagulation enhancers. Examples of coagulation factors include, but are not limited to, Factor VII, thrombin, platelets, and tranexamic acid. These factors can be derived from natural or non-natural sources. In certain embodiments, Factor VII is added to the nanoparticle composition at a concentration of 70-150 IU / kg, thrombin is added to the nanoparticle composition at a concentration of 15-40 IU / kg, and fibrinogen is added to the nanoparticle composition at a concentration of 50-90 mg / kg. Natural or synthetic platelets or platelet substitutes can also be added.

[0139] Antioxidants

[0140] In certain embodiments, the nanoparticle composition can further comprise one or more antioxidants. Examples of antioxidants include, but are not limited to, sodium bisulfite, sodium sulfite, sodium metabisulfite (e.g., sodium metabisulfite), Eulmi white powder (CH2OHSO2Na), ascorbic acid, sodium ascorbate, erythorbic acid, sodium erythorbate, cysteine, cysteine hydrochloride, homocysteine, glutathione, thioglycerol, alpha-thioglycerol, sodium edetate, citric acid, isopropyl citrate, potassium dichloroisocyanurate, sodium mercaptoacetate, sodium pyrosulfite, 1,3-butanediol, calcium disodium edetate, disodium edetate, amino acid sulfite (e.g., L-lysine sulfite), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, ascorbyl palmitate, vitamin E and its derivatives (e.g., dl-alpha-tocopherol, tocopheryl acetate, natural vitamin E, d-delta-tocopherol, mixed tocopherols, and trolox (water-soluble vitamin E)), guaiacwood, nordihydroguaiaretic acid (NDGA), L-ascorbyl stearate, soybean lecithin, ascorbyl palmitate, benzotriazole, pentaerythritol-tetrakis[3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate], 2-mercaptobenzimidazole. Among these, preferred are sodium bisulfite, sodium sulfite, ascorbic acid, homocysteine, dl-alpha-tocopherol, tocopheryl acetate, glutathione, and trolox (water-soluble vitamin E).

[0141] Other ingredients

[0142] In addition to the above ingredients, the nanoparticle composition can further comprise other additives, including but not limited to: antibiotics, such as penicillin, cloxacillin, dicloxacillin, cephalosporin, erythromycin, amoxicillin-clavulanate potassium, ampicillin, tetracycline, trimethoprim-sulfamethoxazole, chloramphenicol, ciprofloxacin, aminoglycosides (e.g., tobramycin and gentamicin), streptomycin, sulfonamides, kanamycin, neomycin, and monobactams; antiviral agents, such as amantadine hydrochloride, rimantadine, acyclovir, famciclovir, foscarnet, ganciclovir sodium, idoxuridine, ribavirin, sorivudine, trifluridine, valacyclovir, valganciclovir, valtorcitabine, vidarabine, didanosine, stavudine, zalcitabine, zidovudine, alpha-interferon, and edoxudine; antifungal agents, such as terbinafine hydrochloride, nystatin, amphotericin B, griseofulvin, ketoconazole, miconazole nitrate, flucytosine, fluconazole, itraconazole, clotrimazole, benzoic acid, salicylic acid, voriconazole, caspofungin, and selenium sulfide; vitamins, amino acids, vasodilators (e.g., alcohols and polyols), surfactants, antibodies against deleterious cytokines (e.g., tumor necrosis factor (TNF) or interleukins), and vasoactive mediators and immunomodulators, such as prostaglandins, leukotrienes, pro-opiomelanocortin fragments, and platelet-activating factor.

[0143] In certain embodiments, the nanoparticle composition can further comprise a beneficial anion, such as lactate or glutamate. It has been found that compositions containing high-osmolar lactate are effective in reducing brain edema in patients with acute hemodynamic abnormalities. In one embodiment, the nanoparticle composition comprises 250 to 2400 mM lactate or lactate salt. In another embodiment, the nanoparticle composition comprises 250 to 2400 mM lactate or lactate salt and 2 to 10 mM potassium.

[0144] In certain other embodiments, the nanoparticle composition can comprise a substituted cation. For example, the nanoparticle composition can comprise choline in place of sodium.

[0145] In still other embodiments, the nanoparticle composition further comprises a potassium channel blocker, which is capable of inhibiting programmed cell death by preventing potassium efflux.

[0146] In certain embodiments, the nanoparticle composition further comprises an anticancer drug and / or an intracellular signaling molecule, such as cyclic adenosine monophosphate (cAMP) and diacylglycerol. In other embodiments, the nanoparticle composition further comprises one or more organelles or components of organelles, such as all or part of the endoplasmic reticulum, ribosomes, and mitochondria.

[0147] In other embodiments, the nanoparticle composition can be combined with red blood cells, modified red blood cells, or other cellular components of blood.

[0148] In other embodiments, the nanoparticle composition further comprises a pro-opiomelanocortin fragment, such as a beta-endorphin, melanocyte stimulating hormone, enkephalin, or an opiate, to modulate immune response and provide analgesic effects. Beta-endorphin can also be used to modulate neutrophil function in sepsis states at a final concentration of 0.01-100 nm, preferably 0.1-10 nm, and more preferably about 1 nm.

[0149] In other embodiments, the nanoparticle composition further comprises one or more neuroactive agents for treating or preventing a psychiatric disorder.

[0150] IV. Preparation of the Nanoparticle Composition

[0151] The nanoparticle composition can be prepared by mixing the lipophilic or hydrophobic component, the emulsifier, the aqueous carrier, and any other ingredients to form an emulsion. Common mixing methods include, but are not limited to, stirring, shaking, homogenization, vibration, microfluidization, and sonication.

[0152] An exemplary homogenizer is an APV 2000 homogenizer (APV Rannie). The emulsion can be formed using a pressure setting of about 15,000 to 20,000 psi for nanoemulsions of < 100 nm, or about 22,000 to 28,000 psi for larger micellar emulsions of about 300 nm. Multiple rounds (cycles) of homogenization can be required to produce micelles of the desired size. The number of homogenization cycles can vary depending on the formulation, for example, 6 cycles, 8 cycles, 10 cycles, 12 cycles, 15 cycles can be required.

[0153] A suitable particle analyzer and / or zeta potential analyzer can be used to evaluate and monitor the particle size and stability of the micellar composition. An exemplary analyzer includes a Malvern Zetasizer Nano ZS, which can provide particle size and zeta potential measurements.

[0154] In one embodiment, the nanoparticle composition is formed by mixing a preformed lipid emulsion from the above ingredients with an aqueous carrier. In addition, the nanoparticle composition can be encapsulated in a red blood cell ghost. Specifically, the emulsion should be prepared in a manner that allows the lipophilic gas to dissolve into the lipophilic or hydrophobic portion of the emulsion, but not form microbubbles that can increase the risk of gas emboli.

[0155] In certain embodiments, 2-40% (w / v), about 2-20% (w / v), about 4-10% (w / v), or about 5% (w / v) of albumin or albumin polymers or albumin polymers conjugated to amino acids or peptides are added to the nanoparticle composition. The albumin or albumin polymers or albumin polymers conjugated to amino acids or peptides are added to the nanoparticle composition after micelle formation. In one embodiment, the lipophilic or hydrophobic components, emulsifiers, aqueous carrier, and any other non-albumin ingredients are mixed to form an emulsion. Subsequently, the albumin, albumin polymers or albumin polymers conjugated to amino acids or peptides are dissolved into the emulsion at the desired concentration.

[0156] In some embodiments, the A portion or the mixture of the A portion and the B portion are loaded with oxygen, nitric oxide, carbon monoxide, xenon, argon, hydrogen sulfide, other hydrophobic gases, or mixtures of these gases prior to use. The use of these gases is to deliver oxygen for aerobic metabolism after the initial dose, to provide an initial dose of carbon monoxide to prevent multiple organ dysfunction syndrome (MODS), to open blood vessels in vascular diseases or conditions involving vasoconstriction or obstruction; to deliver xenon or argon to prevent the effects of traumatic brain injury or seizures; to deliver hydrogen sulfide to promote long-term tissue preservation. Micelles loaded with nitric oxide can also be used as an anti-hypertensive drug. The A portion, the B portion, or the mixture of the A portion and the B portion can all be sterilized by autoclaving.

[0157] In some embodiments, the soybean oil that enhances coagulation is replaced with chia seed oil that is anti-inflammatory and reduces coagulation. In one embodiment, the nanoparticle composition containing soybean oil is used for the initial phase of the infusion where bleeding occurs. When bleeding is no longer a problem, the nanoparticle composition containing chia seed oil is used for the subsequent phase of the infusion.

[0158] In other embodiments, the glycerol in the A portion is replaced with mannitol. In other embodiments, the lecithin is replaced with a-phosphatidylcholine to eliminate a potential source of protein contamination and allergic reactions (lecithin is contaminated with egg proteins). In other embodiments, the amino acids in the B portion of Formulation 2 are replaced with N-acetylated amino acids. In one embodiment, the nanoparticle composition is a non-oxygenated nanoparticle composition. The term "non-oxygenated nanoparticle composition" as used herein means a formulation that is prepared in an atmospheric environment and not loaded with oxygen by any oxygenation device or method.

[0159] In some embodiments, the nanoparticle composition comprises: 6-18% (w / v) of an amphiphilic emulsifier; 15-35% (w / v) of a lipophilic or hydrophobic component; a polar liquid carrier; and one or more electrolytes, wherein the amphiphilic emulsifier forms a lipid microemulsion (LM) having a lipophilic or hydrophobic inner core comprising the lipophilic or hydrophobic component in the polar liquid carrier, and wherein the lipid microemulsion has a particle size ranging from 20-140 nm. In other embodiments, the nanoparticle composition comprises lipid microemulsions having a particle size ranging from 30-140 nm, 30-130 nm, 30-120 nm, 30-100 nm, 30-90 nm, 30-80 nm, 30-70 nm, 40-140 nm, 40-130 nm, 40-120 nm, 40-100 nm, 40-90 nm, 40-80 nm, 50-140 nm, 50-130 nm, 50-120 nm, 50-100 nm, 50-90 nm, 50-80 nm, 50-70 nm, 60-140 nm, 60-130 nm, 60-120 nm, 60-100 nm, 60-90 nm, 60-80 nm, 80-140 nm, 80-130 nm, 80-120 nm, 80-110 nm, 80-100 nm, 100-140 nm, 100-130 nm, 100-120 nm, 100-110 nm, 120-140 nm, 120-130 nm, or 130-140 nm as measured by electron microscopy.

[0160] In other embodiments, the nanoparticle composition further comprises liposomes having a particle size ranging from 1-30 nm, 1-25 nm, 1-20 nm, 1-15 nm, 1-10 nm, 3-30 nm, 3-25 nm, 3-20 nm, 3-15 nm, 3-10 nm, 5-30 nm, 5-25 nm, 5-20 nm, 5-15 nm, 5-10 nm, 7-30 nm, 7-25 nm, 7-20 nm, 7-15 nm, 7-10 nm, 10-30 nm, 10-25 nm, 10-20 nm, 10-15 nm, 15-30 nm, 15-25 nm, 15-20 nm, 20-30 nm, 20-25 nm, or 25-30 nm as measured by electron microscopy.

[0161] In other embodiments, the nanoparticle composition comprises nanoparticles (including micelles and liposomes) having an average particle size of about 70-160 nm, 70-150 nm, 70-140 nm, 70-130 nm, 70-120 nm, 70-100 nm, 70-90 nm, 80-160 nm, 80-150 nm, 80-140 nm, 80-130 nm, 80-120 nm, 80-100 nm, 80-90 nm, 90-160 nm, 90-150 nm, 90-140 nm, 90-130 nm, 90-120 nm, 90-100 nm, 90-98 nm, 92-96 nm, or 95-100 nm as measured by dynamic light scattering.

[0162] In some embodiments, the nanoparticle composition comprises a mixture of lipoid micelles having a particle size of 30-500 nm and liposomes having a particle size of 1-30 nm as measured by electron microscopy.

[0163] In some embodiments, the nanoparticle composition comprises a mixture of lipoid micelles and liposomes, wherein the average particle size of all particles is 5-25 nm, 5-20 nm, 5-15 nm, 5-10 nm, 10-25 nm, 10-20 nm, 10-15 nm, 15-25 nm, 15-20 nm, or 20-25 nm as measured by electron microscopy.

[0164] In some embodiments, the nanoparticle composition comprises about 12% egg yolk lecithin and comprises lipoid micelles having a particle size of about 30-500 nm and liposomes having a particle size of about 1-25 nm as measured by electron microscopy. In some embodiments, the nanoparticle composition comprises about 12% egg yolk lecithin and comprises lipoid micelles having a particle size of about 40-100 nm and liposomes having a particle size of about 7-20 nm as measured by electron microscopy.

[0165] In some embodiments, the nanoparticle composition comprises about 5-35% soybean oil and 0.5-20% egg yolk lecithin and comprises lipoid micelles having a particle size of about 15-800 nm and liposomes having a particle size of about 1-300 nm as measured by electron microscopy.

[0166] In some embodiments, the nanoparticle composition comprises about 5-25% soybean oil and 0.5-1.5% egg yolk lecithin and comprises lipoid micelles having a particle size of about 30-400 nm and liposomes having a particle size of about 1-150 nm as measured by electron microscopy.

[0167] In some embodiments, the nanoparticle composition comprises about 5-25% soybean oil and 0.5-1.5% egg yolk phosphatidylcholine, and comprises lipid-loaded micelles having a particle size of about 30-400 nm and liposomes having a particle size of about 1-150 nm as measured by electron microscopy.

[0168] In some embodiments, the nanoparticle composition comprises about 12% egg yolk phosphatidylcholine, and comprises lipid-loaded micelles having a particle size of about 40-100 nm and liposomes having a particle size of about 7-20 nm as measured by electron microscopy.

[0169] In some embodiments, the nanoparticle composition described above further comprises 1-10%, 1-8%, 1-5%, 1-3%, 1-2%, 5-10%, 2-8%, 2-5%, 2-3%, 3-10%, 3-8%, 3-5%, 5-10%, 5-8%, or 8-10% (w / v) glycerol

[0170] In some embodiments, the nanoparticle composition described above further comprises NaCl at a final concentration of 50-200 mM, 50-150 mM, or 50-100 mM.

[0171] In some embodiments, the nanoparticle composition is prepared in an atmosphere that is not enriched in oxygen, carbon monoxide, nitric oxide, or xenon.

[0172] In certain embodiments, the nanoparticle composition can be loaded with a lipophilic gas prior to clinical use. Examples of such gases include, but are not limited to, oxygen, xenon, argon, nitric oxide, carbon monoxide, hydrogen sulfide. These gases are present in effective concentrations that modulate vascular function and cellular metabolism. As used herein, “nanoparticle composition loaded with a lipophilic gas” refers to a composition that has been processed to increase the content of the lipophilic gas in the nanoparticle composition. The nanoparticle composition can be loaded with a lipophilic gas by bubbling the lipophilic gas through the nanoparticle composition for a desired length of time, or vortexing the nanoparticle composition with the lipophilic gas under pressurized conditions.

[0173] In one embodiment, the nanoparticle composition is oxygenated by bubbling pure oxygen or a gas having an oxygen content of 21% to 100% (v / v), preferably 40% to 100% (v / v), more preferably 60% to 100% (v / v), most preferably 80% to 100% (v / v) through the mixture for more than 30 seconds, preferably 1-15 minutes, more preferably 1-5 minutes. The pressure can also be reduced to 1 atmosphere after the oxygen is injected under pressure. In one embodiment, the nanoparticle composition is oxygenated immediately prior to use. A portable oxygen tank or a portable oxygen generator (e.g., Evergo portable pulse oxygen generator, manufacturer: Philips Medical, Andover, MA) can be used to oxygenate the nanoparticle composition.

[0174] An alternative approach is to allow the emulsion to reach saturated equilibrium with the target gas atmosphere. In most cases, a bubble trap is required to remove bubbles that could potentially cause gas emboli. The equilibration time for a specific nanoparticle composition can be determined experimentally.

[0175] In some embodiments, the nanoparticle composition comprises an oxygenated lipid emulsion. As used herein, the term "oxygenated lipid emulsion" or "oxygenated nanoparticle composition" refers to a specific type of aerated lipid emulsion or aerated liquid that has been saturated with pressurized oxygen to a total dissolved oxygen concentration that exceeds the oxygen content of the liquid at equilibrium with atmospheric conditions. Example

[0176] The following examples are intended to provide those of ordinary skill in the art with a complete technical disclosure and illustration of how to implement the methods of the present application and should not be construed as limiting the scope of the invention. While every effort has been made to ensure the accuracy of the values ​​used (e.g., amounts, temperatures, etc.), experimental errors and deviations should be taken into account. Unless otherwise indicated, parts are by weight, molecular weights are weight-average molecular weights, temperatures are in degrees Celsius, and pressures are at or near atmospheric pressure.

[0177] Example 1: Preparation of Nanoparticle Composition No. 1

[0178] Preparation Part A

[0179] 20% soybean oil = 20g / 100ml

[0180] Egg yolk lecithin 1.2g / 100ml

[0181] Ultrasonication, microfluidization and / or homogenization are performed to produce a nanoparticle composition.

[0182] Preparation Part B

[0183] Glycerol 2.25%

[0184] NaCl 77mM

[0185] Example 2: Preparation of Nanoparticle Composition No. 2

[0186] Preparation Part A

[0187] Soybean oil 10%-20% = 10-20g / 100ml.

[0188] Egg yolk lecithin 0.6% to 12% = 0.6 to 12g

[0189] Ultrasonication, microfluidization and / or homogenization are performed to produce a nanoparticle composition.

[0190] Preparation Part B

[0191] Glycerol 2.25%

[0192] NaCI 77 mM

[0193] Example 3: Preparation of Nanoparticle Composition No. 3

[0194] Preparation Part A

[0195] Soybean oil 10-20% = 10-20 g

[0196] Egg yolk lecithin 0.6% to 12% = 0.6 to 12 g

[0197] Ultrasonic treatment, microfluidization treatment and / or homogenization treatment is performed to produce the nanoparticle composition.

[0198] Preparation Part B

[0199] Glycerol 2.25%

[0200] NaCI 77 mM

[0201] L-Histidine 1 mM

[0202] Example 4: Preparation of Nanoparticle Composition No. 4

[0203] Preparation Part A

[0204] Egg yolk lecithin 0.6% to 12% = 0.6 to 12 g

[0205] Ultrasonic treatment, microfluidization treatment and / or homogenization treatment is performed to produce the nanoparticle composition.

[0206] Preparation Part B

[0207] Glycerol 2.25%

[0208] NaCI 77 mM

[0209] Example 5: Preparation of Nanoparticle Composition No. 5

[0210] Preparation Part A

[0211] Egg yolk lecithin 0.6% to 12% = 0.6 to 12 g

[0212] Ultrasonic treatment, microfluidization treatment and / or homogenization treatment is performed to produce the nanoparticle composition.

[0213] Preparation Part B

[0214] Glycerol 2.25%

[0215] NaCl 77 mM

[0216] L-Histidine 1 mM

[0217] Example 6: Preparation of Nanoparticle Composition No. 6

[0218] Preparation Part A

[0219] Soybean oil 10-20% = 10-20 g

[0220] Egg yolk phospholipid 0.6% to 12% = 0.6 to 12 g

[0221] Ultrasonic, microfluidic, and / or homogenization processing is performed to produce the nanoparticle composition.

[0222] Preparation Part B

[0223] Glycerol 2.25%

[0224] NaCl 77 mM

[0225] L-Histidine 1 mM

[0226] Albumin 5% (w / v)

[0227] Example 7: Preparation of Nanoparticle Composition No. 7

[0228] Preparation Part A

[0229] Soybean oil 10-20% = 10-20 g

[0230] Egg yolk phospholipid 0.6% to 12% = 0.6 to 12 g

[0231] Ultrasonic, microfluidic, and / or homogenization processing is performed to produce the nanoparticle composition.

[0232] Preparation Part B

[0233] NaCl 103 mM

[0234] L-Sodium lactate 34 mM

[0235] L-Histidine 1 mM

[0236] Part A can be used alone, mixed with Part B just prior to use, or pre-mixed 24 hours prior to use. Either part or both parts can also be lyophilized and reconstituted with water at the time of use. In some embodiments, glycerol is added to the final product at a final concentration of 1.13% (w / v) or 2.25% (w / v).

[0237] Example 7: Treatment of hyperprocalcitoninemia using the nanoparticle composition of the present application.

[0238] A septic patient with hyperprocalcitoninemia was treated with the nanoparticle composition of Example 1. Nanoparticle composition treatment resulted in a decrease in the concentration of procalcitonin in the bloodstream and an improvement in multi-organ function. Blood procalcitonin concentration was measured prior to nanoparticle composition treatment and then monitored dynamically over the 48 hour treatment period. Results are shown in Table 1.

[0239] Table 1: Procalcitonin levels in patients treated with the nanoparticle composition of the present application

[0240]

[0241]

[0242] These data show a significant decrease in procalcitonin within 48 hours of infusion of the VBI-S nanoparticle composition. This decrease cannot be attributed entirely to an improvement in the patient's condition, as patients 01-012-002, 01-012-005, and 01-006-003, who did not survive to the end of the 48 hour treatment period, showed a decreasing trend in procalcitonin. This decrease was not due to dilution. Since there was no change in the concentration of sodium and blood urea nitrogen, and the average volume of nanoparticle composition administered was only 632 ml, this decrease was not due to dilution.

[0243] The decrease in procalcitonin induced by the nanoparticle composition of the present application contradicts existing knowledge that infusion of liposome compositions increases the blood concentration of inflammatory mediators such as tumor necrosis factor, interleukin-6, and others that are known to increase procalcitonin. Therefore, one skilled in the art would expect that infusion of the nanoparticle composition of the present application would result in an increase in procalcitonin, but we observed a clear and sustained decrease.

[0244] The terms and descriptions herein are intended to be exemplary only and should not be construed as limiting. One skilled in the art will understand that various changes can be made within the scope of the application as defined by the appended claims and their equivalents, wherein all terms are to be construed in their broadest reasonable meaning unless otherwise specifically defined.

Claims

1. A method for treating hypercalcitoninemia in a subject, the method comprising: administering to the subject in need of treatment an effective amount of a nanoparticle composition comprising: 0.1%-100% (w / v) amphiphilic emulsifier; 0-35% (w / v) lipophilic or hydrophobic component; a polar liquid carrier; and one or more electrolytes, Wherein, the nanoparticle composition comprises liposomes and / or micelles with a particle size of 1-800 nm.

2. The method according to claim 1, wherein The lipophilic or hydrophobic component is selected from the group consisting of soybean oil, chia seed oil and algae oil.

3. The method according to claim 1 or 2, wherein: The amphiphilic emulsifier is selected from the group consisting of phospholipids, α-phosphatidylcholine, amphiphilic peptides and amphiphilic polymers.

4. The method according to any one of claims 1 to 3, wherein: The amphiphilic emulsifier is selected from the group consisting of egg yolk lecithin and soybean lecithin.

5. The method according to any one of claims 1 to 4, wherein: The polar liquid carrier is selected from the group consisting of water, water-based solutions and non-aqueous polar liquids.

6. The method according to any one of claims 1 to 5, wherein: The non-aqueous polar liquid is selected from the group consisting of dimethyl sulfoxide, polyethylene glycol and polar silicone liquid.

7. The method according to any one of claims 1 to 6, wherein: The electrolyte is selected from the group consisting of one or more of the following: sodium chloride, sodium bicarbonate, sodium citrate, sodium lactate, sodium sulfate, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium acetate, sodium glycerophosphate, sodium carbonate, amino acid sodium salts, sodium propionate, sodium hydroxybutyrate, sodium gluconate, potassium chloride, potassium acetate, potassium gluconate, potassium bicarbonate, potassium glycerophosphate, potassium sulfate, potassium lactate, potassium iodide, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium citrate, amino acid potassium salts, potassium propionate, potassium hydroxybutyrate, calcium chloride, calcium gluconate, calcium lactate, calcium glycerophosphate, calcium pantothenate, calcium acetate, magnesium chloride, magnesium sulfate, magnesium glycerophosphate, magnesium acetate, magnesium lactate, amino acid magnesium salts, ammonium chloride, zinc sulfate, zinc chloride, zinc gluconate, zinc lactate, zinc acetate, ferric sulfate, ferric chloride, ferrous gluconate, copper sulfate and manganese sulfate.

8. The method according to any one of claims 1 to 7, wherein: The nanoparticle composition is administered intravenously, intraarterially, intraosseously, or intracardially.

9. The method according to any one of claims 1 to 8, wherein: The nanoparticle composition is an oxygenated nanoparticle composition, and the oxygen content thereof is 1-50,000 ml O2 per 100 ml of the nanoparticle composition.

10. The method according to any one of claims 1 to 9, wherein: The ratio (w / w) of the emulsifier to the lipophilic or hydrophobic component in the nanoparticle composition is about 1:200 to about 1:1.

7.

11. The method according to any one of claims 1 to 10, wherein: The nanoparticle composition comprises micelles and liposomes, wherein the particle size of the micelles in the nanoparticle composition is 15-200 nm as measured by electron microscopy, and wherein the particle size of the liposomes in the nanoparticle composition is 1-25 nm as measured by electron microscopy.

12. The method according to any one of claims 1 to 11, wherein: The nanoparticle composition has a magnesium ion concentration in a subphysiological range.

13. The method according to any one of claims 1 to 12, wherein: The nanoparticle composition further comprises one or more selected from the group consisting of a crystalline agent, a colloidal agent, an anti-inflammatory agent, an immunomodulatory agent, and a lipophilic gas.

14. The method according to any one of claims 1 to 13, wherein: The nanoparticle composition further comprises glycerol.

15. A method of treating a disease or condition associated with elevated procalcitonin in a subject, the method comprising: administering to the subject in need of treatment an effective amount of a nanoparticle composition comprising: 0.1%-100% (w / v) amphiphilic emulsifier; 0-35% (w / v) lipophilic or hydrophobic component; a polar liquid carrier; and one or more electrolytes, Wherein, the nanoparticle composition comprises liposomes and / or micelles with a particle size of 1-800 nm.

16. The method according to claim 15, wherein The disease or condition associated with elevated procalcitonin is chronic or acute renal failure.

17. The method according to claim 15, wherein: A disease or condition associated with elevated procalcitonin is chronic or acute liver failure.

18. The method according to claim 15, wherein Diseases or conditions associated with elevated procalcitonin result from blood loss, sepsis, heart failure, or damage to the nervous system or status epilepticus.

19. The method according to claim 15, wherein The disease or condition associated with elevated procalcitonin is chronic or acute respiratory failure.

20. The method according to claim 15, wherein A disease or condition associated with elevated procalcitonin is sarcopenia of the elderly.

21. The method according to claim 15, wherein A disease or condition associated with elevated procalcitonin is dementia.

22. The method according to claim 15, wherein The disease or condition associated with elevated procalcitonin is orthopedic or non-orthopedic trauma.

23. The method according to claim 15, wherein The disease or condition associated with elevated procalcitonin is surgery.

24. The method according to claim 15, wherein The disease or condition associated with elevated procalcitonin is myocardial infarction.

25. The method according to claim 15, wherein A disease or condition associated with elevated procalcitonin is autism.

26. The method according to claim 15, wherein A disease or condition associated with elevated procalcitonin is ischemic stroke.

27. The method according to claim 15, wherein A disease or condition associated with elevated procalcitonin is Parkinson's disease.

28. The method according to claim 15, wherein A disease or condition associated with elevated procalcitonin is a vasculitis.

29. The method according to claim 15, wherein A disease or condition associated with elevated procalcitonin is bone fracture.

30. The method of claim 15, wherein: A disease or condition associated with elevated procalcitonin is depression.

31. The method according to claim 15, wherein The disease or condition associated with elevated procalcitonin is cancer or cancer metastasis.

32. A method for treating aging, comprising: administering to the subject in need of treatment an effective amount of a nanoparticle composition comprising: 0.1%-100% (w / v) amphiphilic emulsifier; 0-35% (w / v) lipophilic or hydrophobic component; a polar liquid carrier; and one or more electrolytes, Wherein, the nanoparticle composition comprises liposomes and / or micelles with a particle size of 1-800 nm.

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