Compositions and methods for preventing loss of organ function associated with chronic organ disease

Abstract

The present disclosure provides a method for treating or preventing loss of organ function due to chronic organ or tissue diseases by administering to a subject an effective amount of a TLR4 agonist, such as an MPLA-like compound. More specifically, the present disclosure provides a method for preventing loss of renal function due to chronic kidney disease, a method for preventing loss of organ function due to acute stress, and a method for preventing loss of renal function due to acute stress, by administering an effective amount of a TLR4 agonist to a subject. Also provided is a pharmaceutical composition comprising a TLR4 agonist useful for carrying out the above methods.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 341,209, filed May 12, 2022, the entire contents of which are incorporated herein by reference.

Background Art

[0002] Organ damage caused by chronic diseases is a widespread problem in the United States and the world. Organ diseases (e.g., due to chronic inflammation and subsequent fibrosis) are progressive and ultimately result in loss of organ function. Examples of chronic organ diseases include chronic kidney disease (CKD) due to end - stage renal disease, liver diseases such as non - alcoholic steatohepatitis (NASH), osteoarthritis, rheumatoid arthritis, fibrotic lung diseases, heart diseases, pancreatitis, cancer, and irritable bowel syndrome.

[0003] Furthermore, acute organ stress leads to reduced function and inevitable decline and can contribute to chronic organ diseases. One well - known example of this is the acute kidney toxicity (stress) associated with platinum - based chemotherapy, which often leads to the development of chronic kidney disease.

[0004] In addition to steroid treatment or organ transplantation, there is currently no effective therapy for treating chronic organ diseases or preventing loss of function due to acute organ dysfunction.

[0005] Many acute and chronic disease states are caused by persistent inflammation due to the activity of the innate immune system in response to specific stressors. The innate immune system is the front line of our defense against invading pathogens such as toxic chemicals, injury, bacteria, viruses, and fungi, as well as stressors such as underlying diseases. Examples of these chronic disease states include the propagation of chronic kidney disease due to diabetes or the propagation of NASH resulting from inflammation as a result of diabetes or obesity. The innate immune system defends against stress by releasing cytokines and chemokines in response to the stimulation of various families of receptors, including pattern recognition receptors (PRRs) that recognize various pathogenic molecules such as pathogen-associated molecular pattern receptors (PAMPs) and damage-associated molecular pattern receptors (DAMPs). In healthy individuals, cytokines and chemokines are proteins that direct various cellular activities that fight and resolve stress-induced damage. Abnormal cytokine regulation, or an imbalance in normal cytokine responses, can initiate and contribute to existing chronic inflammation that causes and results in organ disease.

[0006] Toll-like receptors (TLRs) are a family of receptors that play a crucial role in initiating the innate immune response by recognizing various molecular patterns associated with pathogens such as bacteria and viruses, as well as proteins that can cause cell and tissue damage.

[0007] Stimulation of Toll-like receptor 4 (TLR4) can activate one of two pathways: 1) the myeloid differentiation primary response 88 (MyD88) pathway, which results in the production of inflammatory cytokines, and 2) the TIR domain-containing adapter-inducing interferon-β (TRIF) pathway, which results in the production of anti-inflammatory defensive cytokines and type I interferons such as interferon-β (Figure 1).

[0008] The formulation of MPLA-like compounds is difficult due to the potential need to form stable micelles to improve the hydrophobicity and bioactivity of the molecules. Multiple formulations have been described, including water-in-oil emulsions, suspensions, nanoparticle suspensions, liposome formulations, and aqueous formulations containing co-surfactants, but all of these suffer from multiple drawbacks such as pain at the injection site, injection site reactions, lack of bioavailability, inability to administer orally, inability to administer parenterally, lack of stability, and / or inability to be utilized due to the need for special equipment during use.

[0009] Compositions of MPLA-like compounds and methods useful for retarding or halting the progression of organ disease or tissue damage resulting from chronic inflammation and chronic fibrosis are disclosed herein. Further, compositions and methods for preventing organ dysfunction resulting from acute stress are disclosed.

Summary of the Invention

[0010] Chronic diseases of organs resulting from chronic inflammation and subsequent fibrosis follow a pattern of permanent and ongoing destruction of living functional cells and subsequent replacement by collagen, a non-functional protein that results in fibrosis (scar tissue) (Wilson). The establishment of fibrosis and subsequent organ death are driven by ongoing inflammatory processes associated with the innate immune response. Redirecting the innate immune response from a pro-inflammatory state to an anti-inflammatory (or non-inflammatory, protective) state would restore the balance of the innate immune response to slow or halt the progressive destruction and scarring of organ tissue and initiate the healing process.

[0011] The present invention contemplates using MPLA-like compounds as a treatment to redirect the innate immune response from a pro-inflammatory state to an anti-inflammatory state to restore more normal levels of function.

[0012] In some embodiments, the present invention provides a method for treating or preventing loss of organ function due to chronic organ disease by administering an effective amount of a toll-like receptor 4 (TLR4) agonist to a subject in need of treatment or prevention of loss of organ function due to chronic organ disease.

[0013] In some preferred embodiments, the chronic organ disease is selected from non-alcoholic fatty liver, non-alcoholic steatohepatitis, osteoarthritis, rheumatoid arthritis, irritable bowel syndrome, fibrotic lung disease, heart disease, and any combination thereof.

[0014] In some embodiments, the method prevents loss of organ function due to chronic organ disease, while in other embodiments, the method treats loss of organ function due to chronic organ disease.

[0015] In other embodiments, the present invention provides a method for treating or preventing loss of renal function due to chronic kidney disease by administering an effective amount of a toll-like receptor 4 (TLR4) agonist to a subject in need of treatment or prevention of loss of renal function due to chronic kidney disease.

[0016] In some embodiments, the method prevents loss of renal function due to chronic kidney disease, while in other embodiments, the method treats loss of renal function due to chronic kidney disease.

[0017] The present invention further provides a method for treating or preventing loss of organ function due to acute stress by administering an effective amount of a TLR4 agonist to a subject in need of treatment or prevention of loss of organ function due to acute stress.

[0018] In certain embodiments, the organ is the kidney. In this and other embodiments, the acute stress is caused by one or more of drug-derived toxicity, chemotherapy-derived toxicity, ischemia, trauma, cancer, or infection. In some embodiments, the loss of organ function is due to chronic inflammation or fibrosis.

[0019] In some embodiments, the method prevents loss of organ function due to acute stress. In this and other embodiments, the method treats loss of organ function due to acute stress.

[0020] In certain embodiments, the TLR4 agonist is an MPLA-like compound such as phosphorylated hexaacyl disaccharide (PHAD), 3-deacylated phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof, more preferably, PHAD or a pharmaceutically acceptable salt thereof.

[0021] In some embodiments, the MPLA-like compound is administered parenterally as an aqueous solution, preferably as an aqueous solution. In other embodiments, the MPLA-like compound is delivered orally as a tablet.

[0022] In some embodiments, the TLR4 agonist selectively stimulates the TIR domain-containing adapter-inducing interferon-β (TRIF) pathway.

[0023] In some embodiments, the TLR4 agonist decreases circulating TGF-β in a subject. In this and other embodiments, the TLR4 agonist increases circulating interleukin-10 (IL-10), circulating hepcidin and / or circulating neutrophil gelatinase-associated lipocalin (NGAL) in the subject.

[0024] The present invention further provides a pharmaceutical composition for treating or preventing organ function loss in a subject in need of treating or preventing organ function loss, comprising a colloidal formulation of a monophosphoryl lipid A (MPLA)-like compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the MPLA-like compound is phosphorylated hexaacyl disaccharide (PHAD), PHAD-504, 3D-(6-acyl)-PHAD, 3D-PHAD, or any combination thereof, or a pharmaceutically acceptable salt thereof. In certain preferred embodiments, the MPLA-like compound is PHAD or a pharmaceutically acceptable salt thereof.

[0025] In some embodiments, the pharmaceutical composition is an aqueous composition. In other embodiments, the pharmaceutical composition is a dry powder.

[0026] In some embodiments, the pharmaceutical composition has an MPLA concentration of about 1 μg / mL to about 10,000 μg / mL.

[0027] In some embodiments, the composition further comprises a stabilizer, preferably the stabilizer is trehalose.

[0028] In some embodiments, the composition comprises micelles having an average diameter or length of about 1 nm to about 1000 nm.

[0029] In some embodiments, the composition comprises a bulking agent selected from one or more of the following: mannitol, trehalose, chitosan, HP-β-cyclodextrin, hydroxypropylmethylcellulose (HPMC), dextran, pea starch, and sucrose. BRIEF DESCRIPTION OF THE DRAWINGS

[0030]

Figure 1

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Figure 10

Mode for Carrying Out the Invention

[0031] Definition "About" and "approximately" generally mean within an acceptable degree of error with respect to the measured amount, taking into account the nature or accuracy of the measurement. Typically, the exemplary degree of error is within 20 percent (%) of a given value or range of values, preferably within 10%, more preferably within 5%. Alternatively, and particularly in biological systems, the terms "about" and "approximately" may mean a value within one order of magnitude of a given value, preferably within 5-fold, more preferably within 2-fold.

[0032] As used herein, "colloid" refers to any liquid or solid composition containing a multimolecular aggregated microstructure having a diameter or length on the scale of 1 nm to 10 nm. Such microstructures include, but are not limited to, micelles, liposomes, vesicles, nanoparticles, microparticles, etc. The microstructure may be spherical, oval, elliptical, flat, or any other arbitrary shape.

[0033] As used herein, "micelle" is a term recognized in the art and refers to colloidal-sized particles that exist in equilibrium with molecules or ions in the solution in which they are formed. It is an aggregate (or supramolecular assembly) of molecules dispersed in a liquid and forms a colloidal suspension (also known as an associated colloid system). A typical micelle in water forms an aggregate with a hydrophilic "head" region in contact with the surrounding solvent and isolates a hydrophobic single-tail region at the center of the micelle.

[0034] As used herein, "liposome" is a term recognized in the art and refers to a spherical vesicle having at least one lipid bilayer. Liposomes can be prepared by disrupting biological membranes (such as by sonication).

[0035] As used herein, "vesicle" is a term recognized in the art and refers to a sac filled with a membranous fluid surrounded by a lipid bilayer.

[0036] As used herein, "nanoparticle" is a term recognized in the art and is typically defined as particles of a substance having a diameter of 1 to 100 nanometers (nm). This term may also be used for larger particles up to 500 nm.

[0037] As used herein, "microparticle" is a term recognized in the art and is defined as particles having a size of 1 to 1000 μm.

[0038] The term "treating" includes prophylactic and / or therapeutic treatment. The term "prophylactic or therapeutic" treatment is recognized in the art and includes administration of one or more subject compositions to a host. When administered prior to the clinical signs of an undesirable condition (e.g., a disease or other undesirable condition in a host animal), the treatment is prophylactic (i.e., it protects the host against the development of the undesirable condition), but when administered after the signs of the undesirable condition, the treatment is therapeutic (i.e., it is intended to reduce, ameliorate, or stabilize the existing undesirable condition or its side effects). Treatment of a respiratory virus infection may include reduction or elimination of symptoms such as runny nose, sneezing, itchy eyes, cough, fatigue, headache, sore throat, or congestion.

[0039] As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound that reduces the occurrence of the disorder or condition in a treated sample compared to an untreated control sample in a statistical sample, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition compared to an untreated control sample.

[0040] "Patient", "subject", or "individual" are used interchangeably and refer to either a human or non-human animal. These terms include mammals such as humans, non-human primates, farm animals (including cows, pigs, etc.), companion animals (e.g., dogs, cats, etc.), and rodents (e.g., mice and rats). In some embodiments, the subject is a human.

[0041] As used herein, the phrase "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, lubricant, binder, carrier, humectant, disintegrant, solvent, or encapsulating material, that would be considered suitable by one of ordinary skill in the art for making a pharmaceutical formulation suitable for administration to a subject. Each excipient must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and must also be "pharmaceutically acceptable" as defined above. Examples of materials that can serve as pharmaceutically acceptable excipients include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; celluloses and their derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; silica, waxes; oils such as corn oil and sesame oil; glycols such as propylene glycol and glycerin; polyols such as sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; and other non-toxic compatible substances commonly used in pharmaceutical formulations.

[0042] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or agent is an amount of the drug or agent that, when administered to a subject, has the intended therapeutic effect. A complete therapeutic effect need not occur upon administration of a single dose and may occur only after administration of a series of doses. Thus, a therapeutically effective amount can be administered in one or more administrations. The exact effective amount required for a subject will depend, for example, on the size, health, and age of the subject, as well as the nature and extent of the condition being treated.

[0043] Activation of the MyD88 signaling pathway by TLR4 stimulation results in the production of inflammatory cytokines including IL-1β, IL-6, TNF-α, and IL-18 (Edilova). Prolonged exposure to these cytokines is associated with the inflammatory process and leads to tissue damage in chronic diseases. MPLA-like compounds signal mainly via the TRIF pathway upon TLR4 stimulation, reducing the production of inflammatory cytokines and producing anti-inflammatory cytokines such as IFN-β, IL-4, IL-10, IP-10, and TGF-β, to correct this dysregulation and slow down or halt the progression of chronic organ diseases. The bias of TRIF mediated by TLR4 has been used to describe the cytokine storm observed in inflammatory diseases, sepsis, and ARDS, probably macrophage activation syndrome, as well as the recent observation of "inflammaging" (Rea, 2018) of increased inflammation observed with increasing age, supporting the potential modification of cellular activities associated with specific conditions related to cytokine dysregulation.

[0044] This inflammatory process is driven by cellular activities initiated in response to inflammatory cytokines. A series of cell communications are set in motion in the complex cascade of the inflammatory process in response to tissue or cell damage. At the initial site of injury, sentinel dendritic cells (DCs) are activated, resulting in the production of multiple cytokines, including IL-12 and IL-18, which then activate natural killer cells (NK cells). These activated NK cells can then release large amounts of highly cytotoxic INF-γ and end in cell death (Zwirner). Chronic DC and NK cell activation induces the production of inflammatory cytokines that create a cytotoxic environment and contribute to the inflammatory state.

[0045] Although not bound by theory, MPLA is not thought to completely stop the production of inflammatory cytokines; rather, MPLA increases host resistance during inflammatory events by attenuating the production of inflammatory cytokines, enabling MPLA to fight infection while improving tissue and organ damage (Watts). The reduction of inflammatory signaling from MPLA bound to TLR4, combined with a minimal impairment of the immune-stimulatory adjuvant effect on T cells (Thompson et al., 2005; Mata-Haro et al., 2007), indicates that MPLA may be safe and effective for use as a monotherapy for certain chronic inflammatory conditions.

[0046] A representative example of an MPLA-like compound, which is also a representative example of the main species in bacterial-derived MPLA, the structure of synthetic phosphorylated hexaacyl disaccharide (PHAD) is shown below:

[0047]

Chemical formula

[0048] A common feature of all MPLA-like compounds is monophosphorylated disaccharide. The degree of acylation can vary in the range from as few as 4 acyl groups to as many as 9 acyl groups. Furthermore, the length of each acyl chain (e.g., the number of carbons) can vary from about 8 to about 20 carbons.

[0049] MPLA-like compounds may be synthetic or biologically derived as described (e.g., derived from the hydrolysis of bacterial cell walls). In certain preferred embodiments, MPLA is selected from phosphorylated hexaacyl disaccharide (PHAD), PHAD-504, 3D-(6-acyl)-PHAD, 3D-PHAD, and any combination thereof. In certain preferred embodiments, MPLA is PHAD.

[0050] In some embodiments of the present invention, loss of organ function associated with chronic organ disease can be treated in a subject by administering an effective amount of a TLR4 agonist. In certain embodiments, the TLR4 agonist is an MPLA-like compound. In certain preferred embodiments, the MPLA-like compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0051] In some preferred embodiments, the TLR4 agonist selectively stimulates the TRIF pathway rather than the MyD88 pathway.

[0052] Chronic Diseases and the Innate Immune System Chronic organ diseases are characterized by a progressive loss of organ function. There are many chronic diseases of vital organs including chronic kidney disease (CKD), non-alcoholic steatohepatitis (NASH), osteoarthritis (OA), rheumatoid arthritis (RA), irritable bowel syndrome (IBS), fibrotic lung disease, heart disease, and the like. Many of these chronic diseases are the result of a single stress or repeated stresses, either internal or external stimuli.

[0053] Chronic diseases of organs due to chronic inflammation and subsequent fibrosis follow a pattern of permanent and ongoing destruction of living functional cells and subsequent replacement by collagen, a non-functional protein that results in fibrosis (scar tissue). The establishment of fibrosis and subsequent organ death are driven by ongoing inflammatory processes associated with the innate immune response. Redirecting the innate immune response from a pro-inflammatory state to an anti-inflammatory (non-inflammatory, or protective) state can slow or halt the progressive destruction and scarring of organ tissue and initiate a healing process.

[0054] The present invention contemplates using MPLA-like compounds as a treatment to redirect the innate immune response from a pro-inflammatory state to an anti-inflammatory state to restore a more normal level of function. Activation of the MyD88 signaling pathway by TLR4 stimulation results in the production of inflammatory cytokines including IL-1β, IL-6, TNF-α, and IL-18 (Edilova). Prolonged exposure to these cytokines is associated with the inflammatory process and results in tissue damage in chronic diseases. MPLA-like compounds signal mainly via the TRIF pathway upon TLR4 stimulation, reducing the production of inflammatory cytokines and producing anti-inflammatory cytokines such as IFN-β, IL-4, IL-10, IP-10, and TGF-β to correct this dysregulation and slow or halt the progression of chronic organ diseases.

[0055] The TLR4-mediated bias of TRIF has been used to describe the cytokine storm observed in inflammatory diseases, sepsis, and ARDS, potentially macrophage activation syndrome, as well as the recent observation of "inflammaging" (Rea, 2018) of increased inflammation observed with aging, supporting the potential modification of cellular activities associated with specific conditions related to cytokine dysregulation.

[0056] This inflammatory process is driven by cellular activities initiated in response to inflammatory cytokines. A series of cell communications are set in motion in the complex cascade of the inflammatory process in response to tissue or cell damage. At the initial site of injury, sentinel dendritic cells (DCs) are activated, resulting in the production of multiple cytokines including IL-12 and IL-18, which then activate natural killer cells (NK cells). These activated NK cells can then release large amounts of highly cytotoxic INF-γ, culminating in cell death (Zwirner). Chronic DC and NK cell activation induces the production of inflammatory cytokines that create a cytotoxic environment and contribute to the inflammatory state.

[0057] The efficient production of inflammatory cytokines depends on the assembly of the inflammasome. The inflammasome is a multi - protein structure formed in the cytoplasm of activated innate immune cells that results in the maturation of the active forms of IL - 1β and IL - 18 from their inactive pro - proteins (Chilton). Once the inflammasome is established, the production of inflammatory cytokines is made possible by the MyD88 pathway.

[0058] The relatively low levels of MyD88 - driven activity observed as a result of MPLA - mediated TLR4 activation are likely due to failure of inflammasome assembly (Embry et al., 2011). Compared to the LPS - mediated response, MPLA produces reduced levels of IL - 1β, IL - 6, and TNF - α (Guo, Chentouh, Watts). These reduced levels of inflammatory cytokines can create a deficiency in the signals required to activate DCs that prevent the production of IL - 12 and IL - 18, and in the signals required to activate NK cells that close the feedback loop of IFN - γ - mediated tissue damage associated with inflammatory diseases.

[0059] In some embodiments, the TLR4 agonists described herein selectively activate the TRIF pathway over the MyD88 pathway. In certain embodiments, the TLR4 agonist has an EC 50 for activating the TRIF pathway and an EC 50 for activating the MyD88 pathway that has a ratio greater than from about 1.1:1 to 100,000:1. In some embodiments, the ratio of these EC 50 is greater than about 1.1:1, greater than about 1.5:1, greater than about 2:1, greater than about 5:1, greater than about 10:1, greater than about 100:1, greater than about 200:1, greater than about 500:1, greater than about 1000:1, greater than about 5,000:1, greater than about 10,000:1, or greater than about 100,000:1. In some embodiments, these EC 50The ratio is in the range of about 1:1 to about 100,000:1, about 1.1:1 to about 50,000:1, about 1.1:1 to about 10,000:1, about 1.1:1 to about 1,000:1, about 1.1:1 to about 100:1, about 1.1:1 to about 10:1, or about 1.1:1 to about 5:1.

[0060] Inflammation is a process necessary for the response to injury. Inflammation aids in combating injury or infection through the infiltration of macrophages, T cells, and B cells, the cellular excavation of pathogens and cellular debris as a result of tissue damage, and ultimately enables the resolution of injury or infection. MPLA does not completely halt the production of inflammatory cytokines; rather, MPLA increases host resistance during inflammatory events through attenuation of the production of inflammatory cytokines, enabling MPLA to combat infection while improving tissue and organ damage (Watts). For example, the small amount of IL-12 produced by DCs treated with MPLA is highly significant when compared to that produced by untreated DCs. These small amounts of IL-12 are not sufficient to fully activate NK cells but are sufficient to induce efficient activation of T cells (Ismaili). The decrease in inflammatory signaling from MPLA bound to TLR4, in combination with a minimal impairment of the immunostimulatory adjuvant effect on the initial clonal expansion of T cells (Thompson et al., 2005; Mata-Haro et al., 2007), can indicate that MPLA is safe and effective for use as a monotherapy for certain chronic inflammatory conditions.

[0061] In some embodiments, loss of organ function associated with chronic organ diseases due to inflammation and fibrosis can be treated in a subject by administering an effective amount of a TLR4 agonist. In some embodiments, the TLR4 agonist is an MPLA-like compound. In some preferred embodiments, the MPLA-like compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0062] Monophosphoryl lipid A (MPLA)-like compounds can selectively stimulate TLR4, activate the TRIF pathway, and result in the production of protective cytokines (Figure 1). These protective cytokines have the ability to shift the immune response from a damaging inflammatory response to a more protective anti-inflammatory response, ultimately restoring the balance and normal function of the innate immune response.

[0063] In other embodiments, long-term loss of organ function resulting from acute inflammation due to internal or external stimuli can be treated in a subject by administering an effective amount of a TLR4 agonist. In some embodiments, the TLR4 agonist is an MPLA-like compound. In some preferred embodiments, the MPLA is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0064] In some preferred embodiments, the origin of the acute inflammation includes one or more of the following non-limiting examples: toxicity from a drug, toxicity from chemotherapy, ischemia, trauma, cancer, and infection.

[0065] Chronic kidney disease (CKD) Kidney diseases are a major public health problem affecting approximately 10% of the population in developed countries (1). Acute kidney injury (AKI) affects 13.3 million people per year and can lead to chronic kidney disease (CKD). Both AKI and CKD are increasing worldwide (2). Progression of chronic kidney injury often leads to end-stage renal disease requiring renal replacement therapy (dialysis or transplantation), resulting in significant morbidity and mortality in affected patients.

[0066] CKD can be initiated and propagated in several ways. One frequently seen condition is high blood glucose associated with diabetes (either type 1 or type 2). High blood glucose is toxic to renal cells, creating stress similar to an inflammatory process, leading to the demise of these cells along with subsequent fibrosis, and ultimately resulting in a progressive loss of renal function over time. High arterial blood pressure is another source of stress that initiates the inflammatory process leading to CKD.

[0067] Other causes of CKD include glomerulonephritis (inflammation of the glomeruli), polycystic kidney disease, autoimmune diseases (such as systemic lupus erythematosus), vesicoureteral reflux (a condition where urine flows back into the kidneys), pyelonephritis, interstitial nephritis (inflammation of the renal tubules), kidney stones, obstruction or cancer of the kidney that can lead to renal failure over the years, excessive use of certain medications, drug abuse (such as heroin or cocaine), and chemotherapy (such as cisplatin).

[0068] Loss of renal function can be measured by several methods known in the art, including, by way of non-limiting example, measurement of estimated glomerular filtration rate or true glomerular filtration rate, measurement of serum creatinine, measurement of blood urea nitrogen, measurement of urinary albumin, and determination of the urinary albumin-to-creatinine ratio.

[0069] In some preferred embodiments of the present invention, the loss of renal function associated with the progression of chronic kidney disease is prevented by administering to the subject an effective amount of a TLR4 agonist. In some preferred embodiments, the TLR4 agonist is an MPLA compound. In a more preferred embodiment, the MPLA compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0070] In other embodiments, the loss of organ function that results from acute inflammation due to acute internal or external stress can be prevented by administering to the subject an effective amount of a TLR4 agonist. In some preferred embodiments, the TLR4 agonist is an MPLA compound. In other preferred embodiments, the MPLA is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0071] In some preferred embodiments, acute stress is an ischemic event such as during hypotensive states (sepsis) such as during kidney surgery or severe infection. Ischemia-induced acute kidney injury (AKI) results in depletion of ATP and changes in epithelial and endothelial cells. This cellular change results in disruption and a decrease in glomerular filtration rate. The cell death induced as a result of AKI is mediated by apoptosis and necrosis. Complex cell interactions direct activities during injury or repair, and certain inflammatory mediators contribute to persistent injury during active renal tubular necrosis (Sharfuddin). In other preferred embodiments, acute stress is due to toxicity as a result of drug overdose or drug abuse. In other preferred embodiments, stress is due to toxicity resulting from chemotherapy. In a preferred embodiment, stress is induced by platinum-containing chemotherapy such as cisplatin. In another preferred embodiment, the acute stress that results in AKI is due to cardiac surgery.

[0072] In surgical situations, such as during procedures that require cardiopulmonary bypass, ischemia can be intentionally initiated. Ischemia can be an unintended consequence of unwanted complications such as intraoperative hypotension. Regardless of the etiology, ischemic events can initially lead away from the affected area where blood, oxygen, and other nutrients are depleted, and then worsen the injury when the blood supply returns to that site along with reactive oxygen species (ROS) and other components that cause oxidative stress to the tissue.

[0073] Ischemic preconditioning and the resulting ischemic tolerance in various organs (e.g., brain, heart, liver, intestine, skeletal muscle) are adaptive defense mechanisms by which sublethal ischemic events or exogenous stimuli (e.g., lipopolysaccharide [LPS] or monophosphoryl lipid A [MPLA]-like compounds) confer resistance to lethal ischemia. Ischemic preconditioning prevents injury upon subsequent exposure to ischemic events by reducing excitotoxicity, apoptosis, and inflammation, thereby protecting mitochondria and increasing antioxidant mechanisms (Bhuiyan 2010). Interventions that can promote ischemic tolerance even in the absence of oxygen depletion techniques are safer modalities for ischemic preconditioning.

[0074] Non-alcoholic steatohepatitis (NASH) Non-alcoholic fatty liver disease (NAFLD) is a condition in which excess fat accumulates in the liver. This accumulation of fat is not caused by excessive alcohol consumption. When excessive alcohol consumption causes fat accumulation in the liver, this condition is called alcohol-related liver disease.

[0075] The two types of NAFLD are non-alcoholic fatty liver (NAFL) and non-alcoholic steatohepatitis (NASH). People typically develop one type of NAFLD or the other, although people with one form may later be diagnosed with the other form of NAFLD.

[0076] NASH is a form of NAFLD that has inflammation and liver injury in addition to fat in the liver. The inflammation and liver injury in NASH can lead to liver fibrosis, or scarring. NASH can result in cirrhosis, in which the liver becomes scarred and permanently damaged. Cirrhosis can lead to liver cancer.

[0077] In some preferred embodiments of the present invention, the loss of liver function associated with the progression of NASH is prevented by administering to the subject an effective amount of a TLR4 agonist. In some preferred embodiments, the TLR4 agonist is an MPLA compound. In a more preferred embodiment, the MPLA compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0078] Osteoarthritis Osteoarthritis is the most common form of arthritis among the elderly and is one of the most frequent causes of physical disability among the elderly. This disease affects both men and women. Before the age of 45, osteoarthritis is more common in men than in women. After the age of 45, osteoarthritis is more common in women. Osteoarthritis occurs when the cartilage, which is the tissue that cushions the ends of the bones within the joint, breaks down and wears away. The inflammatory process drives the breakdown of the cartilage and the joint. In extreme cases, all the cartilage wears away, exposing the bones to rub directly against each other and, if possible, requiring the insertion of an artificial joint.

[0079] In some preferred embodiments of the present invention, the loss of joint function associated with the progression of OA is prevented by administering to the subject an effective amount of a TLR4 agonist. In some preferred embodiments, the TLR4 agonist is an MPLA compound. In a more preferred embodiment, the MPLA compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0080] Rheumatoid arthritis (RA) RA is an autoimmune and inflammatory disease that primarily attacks joints, usually many joints simultaneously. RA generally affects the joints of the hands, wrists, and knees. In joints with RA, the inner side of the joint becomes inflamed, causing damage to joint tissue. This tissue damage can lead to long-lasting or chronic pain, instability (lack of balance), and deformity (malformation). RA can also affect other tissues in the body, causing problems in organs such as the lungs, heart, and eyes.

[0081] In some preferred embodiments of the present invention, the loss of joint function associated with the progression of RA is prevented by administering to the subject an effective amount of a TLR4 agonist. In some preferred embodiments, the TLR4 agonist is an MPLA compound. In a more preferred embodiment, the MPLA compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0082] Irritable bowel syndrome Irritable bowel syndrome (IBS) is a chronic disorder that affects the gastrointestinal tract, causing abdominal pain, bloating, muscle spasms, gas, diarrhea and constipation, or both. Although common, only a small percentage of people with IBS have severe symptoms such as those seen in Crohn's disease and ulcerative colitis, where the persistence of mucosal inflammation has been observed at the microscopic and molecular levels. In certain cases, patients can develop IBS after infection as a result of infectious gastroenteritis, which can contribute to systemic inflammation and perpetuate a cycle of chronic, low-grade asymptomatic inflammation. Upregulated IL-1β has been observed in rectal biopsies of patients with post-infectious IBS. Modifying, or normalizing, cytokine secretion can not only reduce the inflammation contributing to IBS, but also help re-establish a healthy population of gut microbiota.

[0083] In some preferred embodiments of the present invention, reduction of inflammation that may contribute to the progression of IBS or inflammatory bowel disease is prevented by administering to a subject an effective amount of a TLR4 agonist. In some preferred embodiments, the TLR4 agonist is an MPLA-like compound. In more preferred embodiments, the MPLA-like compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD), or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0084] Fibrotic lung disease Fibrotic lung diseases, including idiopathic pulmonary fibrosis, are broad terms used to describe inflammation and scarring (fibrosis) of lung tissue that results in a decline in lung function. Fibrotic lung disease can be caused by multiple factors, including long-term exposure to toxins and pollutants, radiation and radiation treatment, and certain drugs such as chemotherapeutic agents. Drugs designed to kill cancer cells, such as methotrexate (Trexall, Otrexup, etc.) and cyclophosphamide, can also damage lung tissue. Some drugs used to treat arrhythmias, such as amiodarone (Cordarone, Nexterone, Pacerone), can harm lung tissue. Antibiotics such as nitrofurantoin (Macrobid, Macrodantin, etc.) or ethambutol can cause lung damage. Certain anti-inflammatory drugs, such as rituximab (Rituxan) or sulfasalazine (Azulfidine), can cause lung damage. Certain underlying medical conditions (such as dermatomyositis, polymyositis, mixed connective tissue disease, systemic lupus erythematosus, rheumatoid arthritis, sarcoidosis, scleroderma, pneumonia, etc.) may benefit from more effective anti-inflammatory treatment.

[0085] In some preferred embodiments of the present invention, the loss of lung function associated with the progression of fibrotic lung disease is prevented by administering to the subject an effective amount of a TLR4 agonist. In some preferred embodiments, the TLR4 agonist is an MPLA compound. In a more preferred embodiment, the MPLA compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0086] Heart disease Cardiovascular disease (CVD), a class of diseases that affect the heart or cardiovascular system, accounts for 31% of all deaths and remains the leading cause of death worldwide. Ischemic heart disease and endomyocardial fibrosis are the main causes of end-stage heart failure.

[0087] Fibrosis is the leading cause of death and morbidity in heart disease. Fibrotic scarring of the myocardium most commonly occurs after myocardial infarction; however, there are various other conditions that promote cardiac fibrosis such as hypertensive heart disease, diabetic hypertrophic cardiomyopathy and idiopathic dilated cardiomyopathy [4, 5].

[0088] In some preferred embodiments of the present invention, the loss of cardiac function associated with fibrosis of cardiac tissue is prevented by administering to the subject an effective amount of a TLR4 agonist. In some preferred embodiments, the TLR4 agonist is an MPLA compound. In a more preferred embodiment, the MPLA compound is synthetic and is selected from phosphorylated hexaacyl disaccharide (PHAD), 3-deacyl phosphorylated hexaacyl disaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered simultaneously.

[0089] MPLA-like compound The pharmaceutical composition of the present disclosure contains a monophosphoryl lipid A (MPLA)-like compound. MPLA was originally isolated from lipopolysaccharides obtained from the cell walls of Gram-negative bacteria:

[0090]

Chemical formula

[0091] Bacteria-derived MPLA is typically a mixture of several different species and shows one of the main species of bacteria-derived MPLA. By way of example, MPLA may be derived from Salmonella minnesota R595 lipopolysaccharide. As will be understood, MPLA may also be derived from other Salmonella species. Bacterial LPS can be processed by sequential acid and alkali hydrolysis steps to remove polysaccharide side chains, phosphate groups, and partially remove some of the acetyl side chain groups. The crude MPLA can then be purified. The final MPLA product is a mixture of heptaacyl, hexaacyl, and pentaacyl-monophosphorylated glucosamine disaccharide-linked β1a6. Diacetyl, triacetyl, and tetraacetyl, if present, are considered impurities. The acylated lipids vary and include lauroyl, myristoyl, and palmitoyl. The relative ratios of each species may vary between batches, but the main species produced is the hexaacylated disaccharide product.

[0092] The main species found in bacteria-derived MPLA have been chemically synthesized and have immunostimulatory properties equivalent to those of bacteria-derived materials. Examples of synthetic MPLA compounds suitable for use in the present invention include phosphorylated hexaacyl disaccharide (PHAD®) (also known as glucopyranosyl lipid A, or GLA), 3D-PHAD (or 3-acyl-PHAD) (also known as monophosphoryl 3-deacyl lipid A):

[0093]

Chemical formula

[0094] [Chemical formula] include.

[0095] Suitable synthetic variations of MPLA within the scope of the present invention include those in which the fatty acid chain length varies between 10 and 20 carbons and those in which the degree of acylation is penta, hexa, or hepta.

[0096] PHAD is chemically equivalent to the main component of bacterial-derived MPLA. PHAD is also equivalent to bacterial-derived MPLA in terms of biological effects.

[0097] Dosage form and administration route MPLA-like compounds can be administered by several different routes. The choice of route depends on multiple factors including the need for systemic exposure (or lack thereof), the desire to rapidly reach specific organs with the MPLA-like compound, patient acceptability, and compliance.

[0098] Methods of systemic delivery include methods known in the art that result in the delivery of an active molecule (e.g., a drug) to the circulatory system along with distribution throughout the body. Systemic delivery methods include intramuscular, intravenous, subcutaneous, intraperitoneal, sublingual, and oral. As will be understood, any method of systemic delivery is suitable for use with the present invention. Particularly suitable methods of systemic delivery include oral, intramuscular, and intravenous delivery.

[0099] In some embodiments, it may be desirable for the drug to interact only with mucosal tissue and have no or minimal systemic exposure. Methods for mucosal delivery include methods known in the art that result in the delivery of an active molecule to the mucosa. Mucosal delivery methods include intranasal, buccal, sublingual, and oral. A particularly suitable method for mucosal delivery includes intranasal delivery.

[0100] In these embodiments, a composition comprising an MPLA compound can be formulated as droplets, an aerosol, micelles in solution, lipid or liquid nanospheres, liposomes, lipid or liquid microspheres, a solution spray, or a powder for delivery to the nasal cavity or nasal vestibule of a subject. The composition can be administered by direct application to the nasal cavity, or atomized or nebulized for inhalation via the nose or mouth.

[0101] In some embodiments, the method comprises administering a nasal spray, a medicated nasal swab, a medicated wipe, a nasal drop, or an aerosol to the nasal cavity or nasal vestibule of a subject. For this purpose, viscosity modifiers that can be deployed to optimize the product for the application format may include cetyl alcohol, stearyl alcohol, carnauba wax, stearic acid, xanthan gum, aluminum magnesium silicate, gelatin, carbomer, poloxamer, PEG, wax, starch, castor oil derivatives, fatty acids, fatty alcohols, and lecithin.

[0102] In some embodiments, the compositions of the invention can be delivered using a small needleless nasal spray device that allows (self)-administration with little or no prior training to deliver the desired dose. The device may include a reservoir containing a fixed amount of the composition. The device may include a pump spray for delivering one or more fixed doses to the subject's nasal cavity. The device may advantageously be for single-dose use or multiple-dose use. The intended dose can be administered using multiple sprays, for example, one spray in each nostril, for example, two sprays, or as a single spray in one nostril, or the dose can be further designed to vary depending on the patient's weight or maturity. In some embodiments, the nasal drops can be pre-packaged in a pouch or ampoule that can be opened immediately before use and squeezed or ejected into the nasal cavity. In some embodiments, the nasal spray or nasal drops can be achieved by reconstitution of the product powder with an aqueous vehicle at the time of use immediately prior to administration. In some embodiments, the nasal spray or nasal drops can be achieved by reconstitution of a solid formulation powder contained in a suitable delivery device using an aqueous vehicle for some period of time during which the formulation is considered stable in solution form prior to patient administration.

[0103] In certain embodiments, the composition is most preferably suitable for parenteral administration to a mammal, by injection or intravenous infusion, and in some embodiments, the composition may include one or more pharmaceutically acceptable excipients. Suitable excipients include pharmaceutically acceptable buffers, stabilizers, local anesthetics, and the like. The composition can be adapted for direct injection or intravenous infusion, or for addition to an intravenous drip for stepwise infusion, by appropriate use of excipients and packaging and delivery means well known in the art.

[0104] In other embodiments, the present invention provides a pharmaceutical packaging comprising a vial or an ampoule containing an MPLA-like compound in the form of a reconstitutable powder or a solution suitable for injection or infusion, together with instructions for administering the composition to a patient in need thereof. The instructions include, but are not limited to, written and / or pictorial explanations of the active ingredient, instructions for diluting the composition to a concentration suitable for administration, suitable indications, suitable dosing regimens, contraindications, drug interactions, and any adverse side effects noted during the course of clinical trials.

[0105] In an alternative embodiment, the pharmaceutical packaging may include, together with the above instructions, a plastic bag containing 100 ml to 2 L of the pharmaceutical composition of the present invention in a form suitable for intravenous administration.

[0106] In alternative embodiments, the pharmaceutical composition of the present invention may be in a form suitable for oral administration, such as, for example, a syrup or a palatable solution; a form suitable for topical application, such as, for example, a cream or an ointment; or a form suitable for administration by inhalation, such as, for example, a microcrystalline powder or a solution suitable for nebulization. Methods and means for formulating pharmaceutical ingredients for alternative routes of administration are well known in the art, and those skilled in the relevant art would be expected to adapt these known methods to the MPLA-like compounds and formulations described in the present invention.

[0107] The present invention provides a pharmaceutically acceptable composition comprising a therapeutically effective amount of one or more MPLA-like compounds formulated together with one or more pharmaceutically acceptable excipients. The pharmaceutical composition of the present invention can be formulated for administration in solid or liquid form, including forms suitable for oral administration, such as aqueous or non-aqueous solutions or suspensions, tablets, powders, and granules; forms suitable for administration by inhalation, such as aerosols, solutions for spraying, or dry powders; forms suitable for parenteral administration, such as sterile solutions or suspensions; forms suitable for topical application, such as lotions, creams, ointments or sprays; forms suitable for ocular administration; or forms suitable for intravaginal or rectal administration, such as pessaries, suppositories, creams or foams. Preferably, the pharmaceutical preparation is suitable for parenteral administration, more preferably, it is a pyrogen-free solution suitable for intravenous administration.

[0108] If desired, tablets can be made by compression or molding, with one or more accessory ingredients. Compressed tablets can be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surfactants or dispersing agents. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0109] Tablets and other dosage forms of the pharmaceutical composition of the present invention can be scored or prepared using coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical industry, as necessary. For example, they can be formulated to provide modified core sustained release or controlled release using various proportions of hydroxypropyl methylcellulose, other polymer matrices, liposomes and / or microspheres to provide the desired release profile. For example, they can be sterilized by filtration through a bacteria-retaining filter or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may optionally contain opacifying agents and may be of a composition that releases the active ingredient only or preferentially in certain parts of the digestive tract in a delayed manner, as necessary. Examples of embedding compositions that can be used include polymeric substances and waxes. The MPLA-like compounds may also be in microencapsulated form, optionally containing one or more of the excipients described above.

[0110] Liquid dosage forms for oral administration of MPLA-like compounds include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the MPLA-like compound, the liquid dosage form may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifying agents.

[0111] In addition to the inert diluent, the oral composition may also contain adjuvants such as wetting agents, emulsifying agents and suspending agents, sweetening agents, flavoring agents, coloring agents, perfumes and preservatives.

[0112] The suspending agent may contain, in addition to the active compound, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, crystalline cellulose (microcrystalline cellulose), aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.

[0113] In a dry powder formulation suitable for inhalation, the particle size of the particulate drug should be such that it enables inhalation of substantially all of the drug into the lungs upon administration of the aerosol formulation, and thus, desirably, it will be in the range of less than 20 μm, preferably 1 - 10 μm, more preferably 1 - 5 μm. The particle size of the drug can be reduced by conventional means, for example, by grinding or micronization. The aerosol formulation preferably contains 0.5 - 30% w / w of the MPLA-like compound based on the total weight of the formulation.

[0114] The propellant may, if necessary, contain an adjuvant having a higher polarity and / or a higher boiling point than the propellant. Examples of polar adjuvants that can be used include (for example, C 2~6 ) aliphatic alcohols and polyols such as ethanol, isopropanol and propylene glycol, preferably ethanol. Generally, only a small amount of polar adjuvant (for example, 0.05 - 3.0% w / w) may be required to improve the stability of the dispersion. However, the formulations of the present invention preferably do not substantially contain polar adjuvants, especially ethanol. Suitable propellants include trichlorofluoromethane (propellant 11), dichlorodifluoromethane (propellant 12), dichlorotetrafluoroethane (propellant 114), tetrafluoroethane (propellant 134a) and 1,1-difluoroethane (propellant 152a), saturated hydrocarbons such as propane, n-butane, isobutane, pentane and isopentane, and alkyl ethers such as dimethyl ether. Generally, up to 50% w / w of the propellant may contain a volatile adjuvant, for example, 1 - 30% w / w of a volatile saturated C1 - C6 hydrocarbon.

[0115] The aerosol formulation according to the present invention may, if necessary, contain one or more physiologically acceptable surfactants upon administration by inhalation.

[0116] For administration by inhalation, the drug is preferably inhaled as a dry powder from a nebulizer, a metered-dose inhaler, or a dry powder inhaler, using gelatin, plastic or other capsules, cartridges, blister packs and / or strips as required.

[0117] The administration of the agent can be indicated for the treatment of mild, moderate or severe acute or chronic symptoms, or for prophylactic treatment. The exact dose to be administered will depend on the age and condition of the patient, the particular particulate agent used and the frequency of administration, and will ultimately be at the discretion of the attending physician. Typically, the administration will range from once to more than four times daily.

[0118] For use in a dry powder inhaler, the active ingredient can be modified by spray drying or compression to form a powder having suitable flow characteristics. More generally, a diluent or carrier that is non-toxic as a whole and inert to the drug is added. Examples of such carriers are polysaccharides such as starch and cellulose, dextran, lactose, glucose, mannitol, and trehalose. The carrier can be further modified by the addition of surface modifiers, pretreatment to form low-wrinkle particles, flow promoters, and flavor masking or flavor modifying agents.

[0119] The pharmaceutical composition of the present invention suitable for parenteral administration may contain one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, or sterile powders that can be reconstituted into sterile injection solutions or dispersions immediately before use, containing an MPLA-like compound, together with antioxidants, buffers, bacteriostatic agents, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0120] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenolsorbic acid and the like. It is also desirable to include in the composition isotonic agents such as sugars and sodium chloride.

[0121] Examples of pharmaceutically acceptable antioxidants include, but are not limited to, ascorbic acid, cysteine hydrochloride, sodium metabisulfite, sodium sulfite, ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, alpha-tocopherol, and chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid.

[0122] Injectable depot formulations are prepared by forming a matrix in which the subject compound is microencapsulated in a biodegradable polymer such as polylactide-polyglycolide. The rate of drug release can be controlled according to the ratio of the drug to the polymer and the nature of the particular polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injection formulations can also be prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0123] Dosage forms for topical or transdermal administration of the compounds of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The MPLA-like compounds can be mixed, under sterile conditions, with a pharmaceutically acceptable carrier and with preservatives, buffers or propellants that may be required.

[0124] Ointments, pastes, creams and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof, in addition to the active compounds of the present invention. Ophthalmic preparations, eye ointments, powders, solutions, etc. are also intended to be within the scope of the present invention.

[0125] Preparations of the present invention suitable for vaginal administration include pessaries, tampons, creams, gels, pastes, foams or spray preparations containing such carriers as are known in the art to be appropriate. Such preparations can be prepared, for example, by mixing one or more MPLA-like compounds with one or more suitable non-irritating excipients including, for example, cocoa butter, polyethylene glycol, or suppository wax which is solid at room temperature but liquid at body temperature and will thus melt in the rectal or vaginal cavity and release the MPLA-like compound.

[0126] Dosage and regimen of the therapeutic agent In certain embodiments, the MPLA-like compound is administered at a total dose of 0.001 to 100 milligrams, depending on the route of administration (e.g., parenteral, oral) and the target of the treatment (acute vs. chronic disease). In preferred embodiments, the total dose administered is 50 to 1000 micrograms. In particularly preferred embodiments, the total dose is about 100 to 500 micrograms. In other preferred embodiments, the total dose is about 5 to 20 mg. In another preferred embodiment, the total dose is about 50 to 100 mg.

[0127] In other embodiments of the present invention, the MPLA-like compound is administered as a single dose. In other embodiments, the MPLA-like compound is administered multiple times. In the case of multiple doses, the MPLA-like compound can be administered daily, once every two weeks, weekly, or monthly. The exact dosing frequency and dose required at each interval will depend on a number of factors including the type of chronic organ disease being treated, the rate of disease progression, and the patient's tolerance. At the beginning of the treatment phase, the dose and / or dosing frequency may be higher, and both the dose and / or dosing frequency may be gradually decreased as the progression of the disease decreases (absence of disease progression) in order to maintain the steady state overall.

[0128] Compositions and methods of preparation In certain embodiments, the pharmaceutical composition is an aqueous composition. In some such embodiments, the pharmaceutical composition may contain an organic solvent. In certain embodiments, the organic solvent may occupy up to 15% of the total final volume of the formulation solution to be administered. In preferred embodiments, the organic solvent may occupy up to 5% of the total final volume of the formulation solution to be administered.

[0129] In certain embodiments, an organic solvent such as an organic solvent selected from alcohol, glycerin, low molecular weight polyethylene glycol, and low molecular weight poloxamer is miscible with water. In certain preferred embodiments, the organic solvent is an alcohol, for example, methanol, ethanol, isopropanol, t-butanol, preferably ethanol.

[0130] In certain embodiments, the pharmaceutical composition further comprises one or more surfactants. In certain embodiments, the one or more surfactants are selected from carboxymethyl cellulose, dodecyltrimethylammonium bromide (DTAB), n-dodecyl octa(ethylene oxide) (C12E8), n-dodecyl tetra(ethylene oxide) (C12E4), dioctanoyl phosphatidylcholine (C8-lecithin), polyoxyl 35 castor oil, Cremophor EL (CrEL), octaethylene glycol monododecyl ether (C12E8), hexadecyltrimethylammonium bromide (CTAB), polypropylene oxide (PPO), polyethylene oxide (PEO), PEO-poly(D,L-lactic acid-co-caprolactone) (PEO-PDLLA), and sodium dodecyl sulfate (SDS). In certain preferred embodiments, the one or more surfactants are carboxymethyl cellulose.

[0131] In certain embodiments, the pharmaceutical composition may contain excipients, additives, extenders, and mucoadhesives. These may include mannitol, trehalose, cyclodextrin, and hydroxypropylmethylcellulose (HPMC). In preferred embodiments, the excipients HP-β-cyclodextrin and trehalose are used.

[0132] In certain embodiments, the pharmaceutical composition comprises phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), sphingomyelin (including brain sphingomyelin), lecithin, lysophosphatidylcholine, lysophosphatidylethanolamine, cerebroside, dipalmitoylphosphatidylcholine (DAPC), didecanoyl-L-alpha-phosphatidylcholine (DDPC), dioleoylphosphatidylcholine (DEPC), dilauroylphosphatidylcholine (DLPC), dilinoleoylphosphatidylcholine, dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC), dipalmitoylphosphatidylglycerol (DAPG), didecanoyl-L-alpha-phosphatidylglycerol (DDPG), dioleoylphosphatidylglycerol (DEPG), dilauroylphosphatidylglycerol (DLPG), dilinoleoylphosphatidylglycerol, dimyristoylphosphatidylglycerol (DMPG), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylglycerol (DSPG), 1-palmitoyl-2-oleoyl-phosphatidylglycerol (POPG), dipalmitoylphosphatidylethanolamine (DAPE), didecanoyl-L-alpha-phosphatidylethanolamine (DDPE), dioleoylphosphatidylethanolamine (DEPE), dilauroylphosphatidylethanolamine (DLPE), dilinoleoylphosphatidylethanolamine, dimyristoylphosphatidylethanolamine (DMPE), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylethanolamine (DSPE),1-palmitoyl-2-oleoyl-phosphatidylethanolamine (POPE), dipalmitoylphosphatidylinositol (DAPI), didodecanoyl-L-alpha-phosphatidylinositol (DDPI), dioleoylphosphatidylinositol (DEPI), dilauroylphosphatidylinositol (DLPI), dilinoleoylphosphatidylinositol, dimyristoylphosphatidylinositol (DMPI), dioleoylphosphatidylinositol (DOPI), dipalmitoylphosphatidylinositol (DPPI), distearoylphosphatidylinositol (DSPI), 1-palmitoyl-2-oleoyl-phosphatidylinositol (POPI), dipalmitoylphosphatidylserine (DAPS), didodecanoyl-L-alpha-phosphatidylserine (DDPS), dioleoylphosphatidylserine (DEPS), dilauroylphosphatidylserine (DLPS), dilinoleoylphosphatidylserine, dimyristoylphosphatidylserine (DMPS), dioleoylphosphatidylserine (DOPS), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylserine (DSPS), 1-palmitoyl-2-oleoyl-phosphatidylserine (POPS), dipalmitoylsphingomyelin, didodecanoylsphingomyelin, dioleoylsphingomyelin, dilauroylsphingomyelin, dilinoleoylsphingomyelin, dimyristoylsphingomyelin, sphingomyelin, dioleoylsphingomyelin, dipalmitoylsphingomyelin, distearoylsphingomyelin, 1-palmitoyl-2-oleoyl-sphingomyelin, and a phospholipid selected from any combination thereof.

[0133] In certain embodiments, the pharmaceutical composition does not contain, or is substantially free of, salts (e.g., NaCl).

[0134] In certain embodiments, the pharmaceutical composition has an MPLA concentration of about 1 μg / mL to about 10,000 μg / mL. In certain embodiments, the pharmaceutical composition has an MPLA concentration of about 50 μg / mL to about 200 μg / mL. In certain embodiments, the pharmaceutical composition has an MPLA concentration of about 125 μg / mL. In certain embodiments, the pharmaceutical composition has an MPLA concentration of about 250 μg / mL.

[0135] In certain embodiments, the pharmaceutical composition further comprises a stabilizer or stabilizers. In certain preferred embodiments, the stabilizer is trehalose. In other preferred embodiments, the stabilizer is HP-β-cyclodextrin. In some of the most preferred embodiments, both HP-β-cyclodextrin and trehalose are used as stabilizers.

[0136] In certain embodiments, the pharmaceutical composition is a dry powder.

[0137] In certain embodiments, the colloidal or micellar solution comprises particles having an average diameter of about 1 nm to about 1000 nm. In certain preferred embodiments, the particles have an average diameter of about 50 nm to about 500 nm. In other more preferred embodiments, the particles have an average diameter of about 100 nm to about 500 nm. In other embodiments, the particles have an average diameter of about 500 nm to about 1000 nm. In certain embodiments, the particles have an average diameter of about 10 nm to about 200 nm. In preferred embodiments, the particles have an average diameter of about 10 nm to about 100 nm. In certain embodiments, the particles have an average diameter of less than 50 nm. In certain embodiments, the particles have an average diameter of less than 100 nm. In certain embodiments, the particles have an average diameter of less than 150 nm. In certain embodiments, the particles have an average diameter of less than 200 nm. In certain embodiments, the particles have an average diameter of less than 250 nm. In certain embodiments, the particles have an average diameter of less than 500 nm.

[0138] In certain embodiments, the pharmaceutical composition further comprises a mucoadhesive. In certain embodiments, the mucoadhesive is selected from cellulose derivatives, polyacrylates, starch, chitosan, glycosaminoglycans, hyaluronic acid, cellulose derivatives, polyacrylates, and any combination thereof.

[0139] In certain preferred embodiments, the pharmaceutical composition further comprises a pH modifier, an emulsifier, a pH buffer, an isotonicity modifier, a stabilizer, a preservative, a surfactant, a bulking agent, a flavor, or any combination thereof.

[0140] In certain embodiments, the bulking agent is selected from mannitol, trehalose, chitosan, HP-β-cyclodextrin, hydroxypropylmethylcellulose (HPMC), dextran, starch (such as pea starch), and sucrose.

[0141] In certain embodiments, the colloid comprises micelles. In certain embodiments, the colloidal suspension comprises liposomes. In certain embodiments, the colloidal suspension comprises nanoparticles. In certain embodiments, the colloidal suspension comprises microparticles.

[0142] In certain embodiments, a method for preparing a pharmaceutical composition disclosed herein, a. dissolving one or more MPLAs in an organic solvent to form an organic solvent / MPLA solution; b. mixing the organic solvent / MPLA solution with water to form a colloidal formulation comprising one or more MPLAs is provided.

[0143] In certain embodiments, step (a) or (b) is performed under sonication.

[0144] In certain embodiments, the organic solvent and water are present in a volume:volume ratio of from about 1:1500 to about 1:50. In certain embodiments, the organic solvent and water are present in a volume:volume ratio of from about 1:1000 to about 1:100. In certain embodiments, the organic solvent and water are present in a volume:volume ratio of about 1:800.

[0145] In certain embodiments, one or more surfactants are added in step (a) or (b). In some preferred embodiments, the surfactant is carboxymethylcellulose. In another embodiment, one or more phospholipids are added in step (a) or (b). In certain embodiments, the mixture in step (b) is lyophilized. In other embodiments, the mixture in step (b) is spray dried. In some preferred embodiments, the mixture in step (b) is stabilized with trehalose. In certain embodiments, step (a) or step (b) is carried out at an elevated temperature. In certain embodiments, the elevated temperature is from about 30 °C to about 50 °C. In certain preferred embodiments, the elevated temperature is about 40 °C. In certain embodiments, the mixture in step (b) has an MPLA concentration of about 125 μg / mL.

[0146] In certain embodiments, the mixture in step (b) has an MPLA concentration in the range of from about 1 μg / mL to about 1000 μg / mL; from about 20 μg / mL to about 500 μg / mL, from about 100 μg / mL to about 300 μg / mL, about 250 μg / mL, or about 125 μg / mL.

[0147] In certain embodiments, the spray dried powder has an MPLA concentration in the final powder of from 0.25 to 10% w / w MPLA / solid. In certain preferred embodiments, the MPLA concentration in the final powder is from 0.25 to 2% w / w MPLA / solid.

[0148] In certain embodiments, the spray dried powder is reconstituted with water prior to administration to a patient.

[0149] In some embodiments, the MPLA compound is administered in a composition comprising one or more pharmaceutically acceptable excipients. The phrase "pharmaceutically acceptable" is recognized in the art. In certain embodiments, this term refers to compositions, excipients, adjuvants, polymers, and other materials and / or dosage forms that are suitable for use in contact with human and animal tissues within the scope of sound medical judgment, without undue toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0150] For example, a composition comprising the MPLA compound can be formulated for nasal delivery as a dry powder, as an aqueous solution, an aqueous suspension, a colloid, an oil-in-water emulsion, a micelle formulation, or a liposome formulation.

[0151] In some embodiments, the MPLA composition comprises micelles of the MPLA compound. Without being bound by theory, the micelles are thought to enhance the activity of MPLA. The size of the micelles is, in some embodiments, from about 50 nm to about 1000 nm. The size of the micelles can be measured by various techniques including dynamic light scattering (DLS), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Thus, in some embodiments, the size of the micelles is from about 50 nm to about 1000 nm when measured by DLS.

[0152] In certain preferred embodiments, the composition further comprises an organic solvent such as alcohol, glycerin, low molecular weight polyethylene glycol, poloxamer, or any combination thereof. In some embodiments, the organic solvent is miscible with water. In some embodiments, the organic solvent is an alcohol such as methanol, ethanol, isopropanol, or t-butanol, preferably ethanol.

[0153] In some embodiments, the composition further comprises a fatty acid salt, a fatty acid, a phospholipid, or any combination thereof.

[0154] In some embodiments, the composition comprises a phospholipid or a mixture of phospholipids. Examples of phospholipids include, but are not limited to, phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidylinositol (PI), and phosphatidylserine (PS), sphingomyelin (including brain sphingomyelin), lecithin, lysophosphatidylcholine, lysophosphatidylethanolamine, cerebroside, dipalmitoyl phosphatidylcholine (DAPC), didodecanoyl-L-alpha-phosphatidylcholine (DDPC), dioleoyl phosphatidylcholine (DEPC), dilauroyl phosphatidylcholine (DLPC), dilinoleoyl phosphatidylcholine, dimyristoyl phosphatidylcholine (DMPC), dioleoyl phosphatidylcholine (DOPC), dipalmitoyl phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC), dipalmitoyl phosphatidylglycerol (DAPG), didodecanoyl-L-alpha-phosphatidylglycerol (DDPG), dioleoyl phosphatidylglycerol (DEPG), dilauroyl phosphatidylglycerol (DLPG), dilinoleoyl phosphatidylglycerol, dimyristoyl phosphatidylglycerol (DMPG), dioleoyl phosphatidylglycerol (DOPG), dipalmitoyl phosphatidylglycerol (DPPG), distearoyl phosphatidylglycerol (DSPG), 1-palmitoyl-2-oleoyl-phosphatidylglycerol (POPG), dipalmitoyl phosphatidylethanolamine (DAPE), didodecanoyl-L-alpha-phosphatidylethanolamine (DDPE), dioleoyl phosphatidylethanolamine (DEPE), dilauroyl phosphatidylethanolamine (DLPE), dilinoleoyl phosphatidylethanolamine, dimyristoyl phosphatidylethanolamine (DMPE), dioleoyl phosphatidylethanolamine (DOPE), dipalmitoyl phosphatidylethanolamine (DPPE),Distearoyl phosphatidylethanolamine (DSPE), 1-palmitoyl-2-oleoyl-phosphatidylethanolamine (POPE), dipalmitoyl phosphatidylinositol (DAPI), didodecanoyl-L-alpha-phosphatidylinositol (DDPI), dioleoyl phosphatidylinositol (DEPI), dilauroyl phosphatidylinositol (DLPI), dilinoleoyl phosphatidylinositol, dimyristoyl phosphatidylinositol (DMPI), dioleoyl phosphatidylinositol (DOPI), dipalmitoyl phosphatidylinositol (DPPI), distearoyl phosphatidylinositol (DSPI), 1-palmitoyl-2-oleoyl-phosphatidylinositol (POPI), dipalmitoyl phosphatidylserine (DAPS), didodecanoyl-L-alpha-phosphatidylserine (DDPS), dioleoyl phosphatidylserine (DEPS), dilauroyl phosphatidylserine (DLPS), dilinoleoyl phosphatidylserine, dimyristoyl phosphatidylserine (DMPS), dioleoyl phosphatidylserine (DOPS), dipalmitoyl phosphatidylserine (DPPS), distearoyl phosphatidylserine (DSPS), 1-palmitoyl-2-oleoyl-phosphatidylserine (POPS), dipalmitoyl sphingomyelin, didodecanoyl sphingomyelin, dioleoyl sphingomyelin, dilauroyl sphingomyelin, dilinoleoyl sphingomyelin, dimyristoyl sphingomyelin, sphingomyelin, dioleoyl sphingomyelin, dipalmitoyl sphingomyelin, distearoyl sphingomyelin, 1-palmitoyl-2-oleoyl-sphingomyelin, and any combination thereof.

[0155] In certain embodiments, the phospholipid is DPPC, DOPC, cholesterol, or a mixture thereof.

[0156] In certain embodiments, the composition comprising MPLA may contain a pH modifier, a pH buffer, an oil / emulsifier (e.g., squalene), an isotonicity modifier, a stabilizer, a preservative, a detergent, a fragrance, a bulking agent, or a secondary immunostimulant. In some embodiments, the composition is a dry powder comprising a bulking agent.

[0157] Examples of secondary immunostimulants include, for example, gonadocorticoids, deoxycholic acid, vitamin D, and beta-glucan. Suitable buffers include sodium chloride-based or potassium chloride-based solutions, such as phosphate buffered saline, potassium buffered saline, or borate buffered saline. In some embodiments, the buffer may contain a salt, detergent, or carbohydrate that retains MPLA upon drying and aids in re-solubilizing MPLA upon encounter with liquid. Suitable carbohydrates include trehalose, sucrose, glucose, and mannose.

[0158] In some embodiments, the composition further comprises a mucoadhesive. Suitable mucoadhesives include glycosaminoglycans (GAGs) including chondroitin sulfate, hyaluronic acid, cellulose derivatives, HP-β-cyclodextrin, polyacrylate, starch, HPMC and any combination thereof.

[0159] In some embodiments, the mucoadhesive is present in the composition in an amount of about 0.1 to about 50 wt%, in the range of about 25 to about 50 wt%, or about 49 wt%.

[0160] In some embodiments, the composition further comprises a sugar. Examples of sugars that can be used in the methods provided herein include, but are not limited to, sucrose, glucose, fructose, lactose, maltose, mannose, galactose, trehalose, and combinations thereof. In certain embodiments, the sugar content is about 49 wt%.

[0161] In some embodiments, the composition is an aqueous liquid. In such embodiments, the concentration of the MPLA compound in the composition may be from about 1 μg / mL to about 1000 μg / mL, from about 20 μg / mL to about 500 μg / mL, from about 100 μg / mL to about 300 μg / mL, or about 250 μg / mL.

[0162] In certain embodiments, the formulation may contain an ionic or non-ionic surfactant. Suitable surfactants include poloxamer 407, poloxamer 181, dodecyltrimethylammonium bromide (DTAB), n-dodecyl octaethylene oxide (C12E8), n-dodecyl tetraethylene oxide (C12E4), and dioctanoyl phosphatidylcholine (C8-lecithin), polyoxyl 35 castor oil, Cremophor EL (CrEL), octaethylene glycol monododecyl ether (C12E8), hexadecyltrimethylammonium bromide (CTAB), polypropylene oxide (PPO), polyethylene oxide (PEO), PEO-poly(D,L-lactic acid-co-caprolactone) (PEO-PDLLA), and sodium dodecyl sulfate (SDS), and any combination thereof.

[0163] In some embodiments, the MPLA formulation has a pH of from about 4 to about 9. In certain preferred embodiments, the pH is from 5 to 8.

[0164] In certain embodiments, the formulation may not contain, or may be substantially free of, phospholipids, surfactants, salts (e.g., NaCl), and / or buffers. Substantially free means that the substance in question occupies less than 0.5 wt%, less than 0.25 wt%, less than 0.1 wt%, less than 0.05 wt%, less than 0.01 wt%, or less than 0.005 wt% of the composition.

[0165] In some embodiments of the present invention, the composition comprises an MPLA compound at a concentration of 1 to 8000 μg / mL. In certain preferred embodiments, MPLA is present at a concentration of 20 to 500 μg / mL. In certain more preferred embodiments, the concentration is 100 to 300 μg / mL. In certain embodiments, the concentration is 250 μg / mL, and in yet other embodiments, the concentration is 125 μg / mL.

[0166] In some embodiments of the present invention, the solution is formulated such that the surfactant is included at a concentration of 1 to 40% w / w, where the surfactant can enhance the absorption of the drug upon administration by preventing degradation / metabolism, enhancing barrier permeability by transient opening of tight junctions, disrupting and holding lipid bilayer filling / complexation / carriage / ion pair formation, and slowing mucociliary clearance. In certain preferred embodiments, the surfactant concentration is 1 to 25% w / w. In certain most preferred embodiments, the surfactant concentration is 15% w / w. Examples of surfactants of interest include, but are not limited to, dipalmitoylphosphatidylcholine, soy lecithin, phosphatidylcholine, sodium taurocholate, sodium deoxycholate, glycodeoxycholic acid, palmitic acid, stearic acid, and oleic acid.

[0167] In some embodiments of the present invention, the composition comprises a mucoadhesive at a concentration of 0.1 to 50% w / w, which can enhance the absorption of the drug upon administration by enhancing retention time and / or slowing mucociliary clearance. In certain preferred embodiments, the mucoadhesive is included at a concentration of 40 to 50% w / w. In certain most preferred embodiments, the mucoadhesive is included at a concentration of 49% w / w. Examples of mucoadhesives of interest include, but are not limited to, cellulose derivatives, HP-β-cyclodextrin polyacrylate, starch, and chitosan.

[0168] In other preferred embodiments, the composition is a powder comprising an MPLA-like compound and one or more bulking agents. Useful compositions comprise from 2.5 to 50 wt% MPLA-like compound and from 50 to 97.5 wt% of one or more bulking agents. Preferred compositions comprise from 5 to 20 wt% MPLA-like compound and from 80 to 95 wt% of one or more bulking agents. Particularly preferred compositions comprise about 10 wt% MPLA-like compound and about 90 wt% of one or more bulking agents.

[0169] Combination therapy A variety of anti-inflammatory treatments are available for specific inflammation-related indications, such as Humira, a monoclonal antibody therapeutic agent that targets TNF-α and removes it from circulation. Since TNF-α is a contributing factor in inflammation, a decrease in TNF-α reduces symptoms associated with several inflammatory conditions such as rheumatoid arthritis, eczema, and psoriasis. Since Humira targets a single, specific inflammatory cytokine, it does not provide a broad range of relief from other sources of inflammation. Monoclonal antibody treatments can take several weeks to show symptom reduction.

[0170] Monoclonal antibody therapy for the treatment of inflammation can provide benefits to the treatment when administered with MPLA. This is because, theoretically, it results in a reduction of symptoms both in the short term and the long term as a result of inflammation. In some embodiments, the monoclonal antibody is directed to target galectin-3. In some embodiments, the monoclonal antibody is directed to target inflammatory cytokines including, but not limited to, IL-6, IL-23, IL-33, and / or IL-1β.

[0171] In some embodiments of the present invention, the composition is administered before, simultaneously with, or after monoclonal antibody treatment. (Examples)

[0172] To more fully understand the invention described herein, the following examples are provided. The examples described in this application are provided to illustrate the compounds, compositions, materials, devices, and methods provided herein and should not be construed as limiting the scope in any way.

Example

[0173] Preparation of PHAD Micelles in 5% Ethanol 1 mg of PHAD was wetted in 0.4 mL of 95% ethanol for 1 minute and then sonicated at 40 °C for 15 minutes until a clear solution was formed. The solution was removed from the sonication bath and QS to 8 mL with water to obtain a uniform formulation of PHAD micelles with a size of 150 nm or smaller. The formulation was used as a liquid or lyophilized product.

Example

[0174] Preparation of PHAD Powder 6 mg of PHAD was wetted and dissolved in 3 mL of 95% ethanol at 40 °C and sonicated at 40 °C for 20 minutes until a clear solution of 2 mg / mL PHAD was obtained. Then, 17 mL of water at 40 °C was added and sonicated to thoroughly mix the bulk solution to obtain a uniform formulation of PHAD micelles with a size of about 150 nm or smaller. 147 mg of HP-β-cyclodextrin and 147 mg of trehalose dihydrate were added to the solution under mixing. The solution was then spray-dried to obtain a final powder of 2% w / w PHAD: 49% w / w HP-β-cyclodextrin: 49% w / w trehalose.

[0175] This powder is stable at ambient temperature over a long period of time and readily dissolves in purified water up to a concentration of about 5 - 10 mg / mL. This powder can be reconstituted for parenteral administration, for example, by intravenous, subcutaneous, or intramuscular routes of administration, or by nasal administration. Alternatively, this same powder can be formulated with additional excipients to form tablets or gels for oral delivery.

Example

[0176] The formulations prepared in Examples 1 and 2 show robust upregulation of IP-10 as a result of TLR4 stimulation in vitro in mouse macrophages (Figure 2). IP-10 is an important cytokine associated with stimulation of the TIRF pathway. Evidence of IP-10 upregulation is confirmation that the PHAD preparations described in this application can selectively stimulate the TRIF pathway, which is further confirmation of improved or reduced activity mediated by the MyD88 signaling pathway, which aids in the improvement of inflammatory cytokine production regarding MPLA as a treatment for inflammatory conditions.

Example

[0177] In validated preclinical models of acute kidney injury (AKI) and chronic kidney disease (CKD), daily administration of REVTx-300, an MPLA-like compound, caused a significant decrease in fibrosis and circulating transforming growth factor-β (TGF-β) in a dose-dependent manner compared to the positive control group. Composite data represent the average of three anatomically different depths (10 images / depth / rat / group = approximately 60 - 65% of the renal cortical area). Renal cortical fibrosis, represented as collagen volume fraction (CVF; quantification of PSR-stained tissue sections), was increased in vehicle-treated UUO-obstructed kidneys compared to sham-operated controls. SB-525334 attenuated the increase in renal cortical CVF induced by UUO. Doses of 0.3 mg / kg and 0.9 mg / kg of REVTx-300 (PHAD, and MPLA-like compound) attenuated the increase in renal cortical CVF induced by UUO. Furthermore, REVTx-300 significantly increased circulating anti-inflammatory interleukin-10 (IL-10) (Figure 8), hepcidin (Figure 6), and neutrophil gelatinase-associated lipocalin (NGAL) (Figure 5) in all groups in a dose-dependent manner compared to the positive control group in the unilateral ureteral obstruction (UUO) model.

[0178] TGF-β is an important promoter of fibrotic growth and directly contributes to collagen accumulation through overproduction of the extracellular matrix. IL-10 is characterized as an anti-inflammatory cytokine because of its ability to reduce the production of inflammatory mediators. Hepcidin and NGAL sequester iron and prevent iron-mediated reactive oxygen tissue damage. There was no significant increase in markers of inflammation (no increase in IL-6, and IL-1β (Figure 9) and IL-18 (Figure 10) increased relatively slightly). These results provide mechanistic evidence for the reduction of fibrosis observed in the UUO model in response to treatment with REVTx-300.

[0179] References Bhuiyan MI, Kim YJ. Mechanisms and prospects of ischemic tolerance induced by cerebral preconditioning. Int Neurourol J. 2010;14(4):203-212. doi:10.5213 / inj.2010.14.4.203 Chentouh R, Fitting C, Cavaillon JM. Specific features of human monocytes activation by monophosphoryl lipid A. Sci Rep. 2018;8(7096). Chilton PM, Embry CA, Mitchell TC. Effects of differences in lipid A structure on TLR4 pro-inflammatory signaling and inflammasome activation. Front Immunol. 2012;3(154). da Silva L, Neves BM, Moura L, Cruz MT, Carvalho E. Neurotensin downregulates the pro-inflammatory properties of skin dendritic cells and increases epidermal growth factor expression. Biochem Biophys Acta. 2011;1813(10):1863-71. Edilova MI, Akram A, Abdul-Sater AA. Innate immunity drives pathogenesis of rheumatoid arthritis. Biomed J. 2021;44(2):172-182. Elenkov IJ, Iezzoni DG, Daly A, Harris AG. Cytokine dysregulation, inflammation, and well-being. Neuroimmunomodulation. 2005;12(5):255-69. Fulop T, Larbi A, Dupuis G, et al. Immunosenescence and inflamm-aging as two sides of the same coin: Friends or foes? Front Immunol. 2018;8(1960). Gaekwad J, Zhang Y, Zhang W, Reeves J, Wolfert MA, Boons GJ. Differential induction of innate immune responses by synthetic lipid A derivatives. J Biol Chem. 2010;285(38):29375-86. Gansevoort RT, Correa-Rotter R, Hemmelgarn BR, Jafar TH, Heerspink HJ, Mann JF, et al. Chronic kidney disease and cardiovascular risk: epidemiology, mechanisms, and prevention. Lancet. (2013) 382:339-52. 10.1016 / S0140-6736(13)60595-4 Guo J, Liu Z, Zhang D, et al. TLR4 agonist monophosphoryl lipid A alleviated radiation-induced intestinal injury. J Immunol Res. 2019;2019:2121095. Ismaili J, Rennesson J, Aksoy E, et al. Monophosphoryl lipid A activates both human dendritic cells and T cells. J Immunol. 2002;168:926-32. Jain A, Kaczanowska S, Davila E. IL-1 receptor-associated kinase signaling and its role in inflammation, cancer progression, and therapy resistance. Front Immunol. 2014;5(553). Mata-Haro, Veronica et al. “The vaccine adjuvant monophosphoryl lipid A as a TRIF-biased agonist of TLR4.” Science (New York, N.Y.) vol. 316,5831 (2007): 1628-32. doi:10.1126 / science.1138963. Mian MF, Lauzon NM, Andrews DW, Lichty BD, Ashkar AA. FimH can directly activate human and murine natural killer cells via TLR4. Mol Ther. 2010;18(7):1379-88. Moura LIF, Silva L, Leal EC, Tellechea A, Cruz MT, Carvalho E. Neurotensin modulates the migratory inflammatory response of macrophages under hyperglycemic conditions. Biomed Res Int. 2013; 2013:941764. Ng QX, Soh AYS, Loke W, Lim DY, Yeo WS. The role of inflammation in irritable bowel syndrome (IBS). J Inflamm Res. 2018;11:345-349. Published 2018 Sep 21. doi:10.2147 / JIR.S174982 O’Connor G, Hart OM, Gardiner CM. Putting the natural killer cell in its place. Immunology. 2006;117(1):1-10. Owen AM, Fults JB, Patil NK, Hernandez A, Bohannon JK. TLR agonists as mediators of trained immunity: Mechanistic insight and immunotherapeutic potential to combat infection. Front Immunol. 2021;11:622614. Pifferi C, Fuentes R, Fernandez-Tejada A. Natural and synthetic carbohydrate-based vaccine adjuvants and their mechanisms of action. Nature Reviews Chemistry. 2021;5:197-216. Sharfuddin, A., Molitoris, B. Pathophysiology of ischemic acute kidney injury. Nat Rev Nephrol 7, 189-200 (2011). https: / / doi.org / 10.1038 / nrneph.2011.16 Thompson Journal of Leukocyte Biology Volume 78, December 2005 doi: 10.1189 / jlb.0305172. Watts BA, Tamayo E, Sherwood ER, Good DW. Monophosphoryl lipid A pretreatment suppresses sepsis - and LPS-induced proinflammatory cytokine production in the medullary thick ascending limb. Am J Physiol Renal Physiol. 2020;319:F8-F18. Wilson MD. Fibrogenesis: Mechanisms, Dynamics and Clinical Implications. Iran J Pathol. 2015;10(2):83-88. Xie Y, Bowe B, Mokdad AH, Xian H, Yan Y, Li T, et al. . Analysis of the global burden of disease study highlights the global, regional, and national trends of chronic kidney disease epidemiology from 1990 to 2016. Kidney Int. (2018) 94:567-81. 10.1016 / j.kint.2018.04.011 Zwirner Norberto Walter, Ziblat Andrea. Regulation of NK Cell Activation and Effector Functions by the IL-12 Family of Cytokines: The Case of IL-27. Frontiers in Immunology. VOL 8, 2017 https: / / www.frontiersin.org / article / 10.3389 / fimmu.2017.00025. DOI=10.3389 / fimmu.2017.00025

Claims

1. (i) loss of organ function due to chronic organ disease, (ii) Loss of organ function due to acute stress Use of Toll-like receptor 4 (TLR4) agonists in the manufacture of pharmaceuticals for the treatment of [condition].

2. The use according to claim 1, wherein the TLR4 agonist is an MPLA-like compound.

3. The use according to claim 1, wherein the TLR4 agonist is phosphorylated hexaacyl dissaccharide (PHAD), 3-deacyl phosphorylated hexaacyl dissaccharide (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl dissaccharide (3D(6-acyl)PHAD), or any combination thereof or a pharmaceutically acceptable salt thereof.

4. The use according to claim 3, wherein the TLR4 agonist is PHAD or a pharmaceutically acceptable salt thereof.

5. The use according to claim 1, wherein the pharmaceutical is administered as an aqueous solution.

6. The use according to claim 1, wherein the pharmaceutical product is delivered parenterally as an aqueous solution.

7. The use according to claim 1, wherein the pharmaceutical is a tablet that can be administered orally.

8. The use according to claim 1, wherein the TLR4 agonist selectively stimulates the TIR domain clustering adapter-induced interferon-β (TRIF) pathway.

9. The use of the TLR4 agonist according to claim 1, wherein the TLR4 agonist reduces circulating TGF-β.

10. The use according to claim 1, wherein the TLR4 agonist increases circulating interleukin-10 (IL-10), circulating hepcidin, and / or circulating neutrophil gelatinase-associated lipocalin (NGAL).

11. The use according to claim 1, wherein the chronic organ disease is selected from non-alcoholic fatty liver, non-alcoholic steatohepatitis, osteoarthritis, rheumatoid arthritis, irritable bowel syndrome, fibrous lung disease, heart disease, chronic kidney disease, pancreatitis, cancer, interstitial nephritis, Crohn's disease, ulcerative colitis, scleroderma, systemic lupus erythematosus, type 2 diabetes, inflammatory aging, psoriasis, mixed connective tissue disease, and any combination thereof.

12. The use according to claim 1, wherein the chronic organ disease is related to chronic inflammation.

13. The use according to any one of claims 1 to 12, wherein the pharmaceutical is for the treatment of loss of renal function due to chronic kidney disease.

14. Loss of kidney function includes the following: Type 1 or type 2 diabetes, hypertension, glomerulonephritis, polycystic kidney disease, autoimmune disease, vesicoureteral reflux, pyelonephritis, interstitial nephritis, kidney stones, kidney obstruction, cancer, drug abuse, nephritis, nephropathy, drug overuse, or chemotherapy The use according to claim 13, relating to one or more of the following.

15. The use according to any one of claims 1 to 12, wherein the pharmacopoeia is for the treatment of loss of organ function caused by acute stress.

16. The use according to claim 15, wherein acute stress is related to acute inflammation.

17. The use according to claim 15, wherein the organ is the kidney.

18. Acute stress includes the following: The use according to claim 15, resulting from one or more of the following: toxicity derived from the drug, toxicity derived from chemotherapy, ischemia, trauma, cancer, infection, pancreatitis, acute kidney injury, acute respiratory distress syndrome, cytokine storm syndrome, interstitial nephritis, dermatomyositis, polymyositis, scleroderma, mixed connective tissue disease, or sepsis.

19. The use according to any one of claims 1 to 12, wherein the loss of organ function is due to fibrosis.

20. A pharmaceutical composition for treating or preventing organ loss in subjects requiring treatment or prevention of organ loss, comprising a colloidal formulation of a monophosphoryl lipid A (MPLA)-like compound.

21. The pharmaceutical composition according to claim 20, wherein MPLA is selected from phosphorylated hexaacyl dissaccharide (PHAD), PHAD-504, 3D-(6-acyl)-PHAD, 3D-PHAD, and any combination thereof.

22. The pharmaceutical composition according to claim 21, wherein the MPLA-like compound is PHAD.

23. The pharmaceutical composition according to claim 20, which is an aqueous composition.

24. The pharmaceutical composition according to claim 20, which is a dried powder.

25. The pharmaceutical composition according to claim 23, having an MPLA concentration of approximately 1 μg / mL to approximately 10,000 μg / mL.

26. The pharmaceutical composition according to claim 20, further comprising a stabilizer.

27. The pharmaceutical composition according to claim 26, wherein the stabilizer is trehalose.

28. The pharmaceutical composition according to claim 20, comprising micelles having an average diameter or length of about 1 nm to about 1000 nm.

29. The pharmaceutical composition according to claim 20, comprising a volume extender selected from one or more of the following: mannitol, trehalose, chitosan, HP-B-cyclodextrin, hydroxypropyl methylcellulose (HPMC), dextran, pea starch, and sucrose.

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