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

Abstract

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

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 341,209, filed on May 12, 2022, the contents of which are hereby incorporated by reference in their entirety.

[0003] background

[0004] Organ damage, due to chronic diseases, is a common problem in the United States and around 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) through end-stage renal disease, liver diseases such as nonalcoholic steatohepatitis (NASH), osteoarthritis, rheumatoid arthritis, pulmonary fibrosis, heart disease, pancreatitis, cancer, and irritable bowel syndrome.

[0005] Furthermore, acute organ stress can lead to decreased functionality and eventual decline, resulting in chronic organ disease. A well-known example of this is the acute renal toxicity (stress) associated with platinum-based chemotherapy, which often leads to the development of chronic kidney disease.

[0006] Aside from steroid therapy or organ transplantation, there are currently no effective therapies for treating chronic organ disease or for preventing loss of function due to acute organ dysfunction.

[0007] Many acute and chronic disease states are caused by permanent inflammation caused by the activity of the innate immune system in response to specific stressors. The innate immune system is our first line of defense against toxic chemicals, injuries, invasive pathogens (such as bacteria, viruses and fungi) and underlying diseases. Examples of these chronic disease states include the spread of chronic kidney disease caused by diabetes or the spread of NASH due to inflammation as a result of diabetes or obesity. The innate immune system releases cytokines and chemokines to resist stress in response to stimulation of various receptors (including pattern recognition receptors (PRRs) that recognize a series of pathogen 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 different cell activities to prevent or resolve stress-induced damage. Cytokine dysregulation or imbalance of normal cytokine responses may not only initiate and cause chronic inflammation, but may also contribute to the existence of chronic inflammation leading to organ disease.

[0008] Toll-like receptors (TLRs) are a family of receptors that play a crucial role in initiating innate immune responses by recognizing distinct molecular patterns associated with pathogens, such as bacteria and viruses, and proteins that may cause cell and tissue damage.

[0009] Stimulation of Toll-like receptor 4 (TLR4) can activate one of two pathways: 1) the myeloid differentiation primary response 88 (MyD88) pathway, which leads to the production of pro-inflammatory cytokines; and 2) the TIR domain clustering adaptor-induced interferon-β (TRIF) pathway, which leads to the production of anti-inflammatory protective cytokines and type I interferons such as interferon β ( Figure 1 ).

[0010] The formulation of MPLA-like compounds is difficult due to the hydrophobicity of the molecules and the potential need to form stable micelles to impart enhanced biological activity. Although a variety of formulations have been described (including oil-in-water emulsions, suspensions, nanoparticle suspensions, liposomal formulations, and aqueous formulations with co-surfactants), these formulations all suffer from a variety of disadvantages (including injection site pain, injection site reactions, lack of bioavailability, inability to orally administer, inability to parenterally administer, lack of stability, and / or lack of practicality due to the requirement for specialized equipment for use).

[0011] Disclosed herein are compositions and methods of MPLA-like compounds useful for slowing or stopping the progression of organ disease or tissue damage as a result of chronic inflammation and chronic fibrosis. In addition, compositions and methods for preventing organ dysfunction due to acute stress are disclosed. SUMMARY OF THE INVENTION

[0013] Chronic disease of organs (due to chronic inflammation and subsequent fibrosis) follows the following pattern: permanent and ongoing destruction of living, functional cells and subsequent replacement by non-functional proteins, collagen, resulting in fibrosis (scar tissue) (Wilson). The establishment of fibrosis and the subsequent death of organs is driven by a persistent inflammatory process 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 will rebalance the innate immune response to slow or interrupt the progressive destruction and scarring of organ tissue, allowing the healing process to occur.

[0014] The present invention contemplates the use of MPLA-like compounds as therapeutics to redirect the innate immune response from a pro-inflammatory state to an anti-inflammatory state to restore more normal levels of function.

[0015] In some embodiments, the present invention provides methods 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 thereof.

[0016] In some preferred embodiments, the chronic organ disease is selected from non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, osteoarthritis, rheumatoid arthritis, irritable bowel syndrome, pulmonary fibrosis, heart disease, and any combination thereof.

[0017] In some embodiments, the methods prevent loss of organ function due to chronic organ disease, while in other embodiments, the methods treat loss of organ function due to chronic organ disease.

[0018] In other embodiments, the invention provides methods for treating or preventing loss of renal function due to chronic kidney disease by administering to a subject in need thereof an effective amount of a Toll-like receptor 4 (TLR4) agonist.

[0019] In some embodiments, the methods prevent loss of kidney function due to chronic kidney disease, while in other embodiments, the methods treat loss of kidney function due to chronic kidney disease.

[0020] The present invention also provides methods 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 thereof.

[0021] In certain embodiments, the organ is a kidney. In this and other embodiments, the acute stress is due to one or more of the following: toxicity from a drug, toxicity from chemotherapy, ischemia, trauma, cancer, or infection. In some embodiments, the loss of organ function is due to chronic inflammation or fibrosis.

[0022] In some embodiments, the methods prevent loss of organ function due to acute stress. In this and other embodiments, the methods treat loss of organ function due to acute stress.

[0023] In certain embodiments, the TLR4 agonist is an MPLA-like compound, such as a phosphorylated hexaacyl disaccharide (PHAD), a 3-deacylated phosphorylated hexaacyl disaccharide (3D-PHAD), a 3D-(6-acyl)phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or a pharmaceutically acceptable salt thereof, and more preferably, PHAD or a pharmaceutically acceptable salt thereof.

[0024] In some embodiments, the MPLA-like compound is administered as an aqueous solution, preferably parenterally as an aqueous solution. In other embodiments, the MPLA-like compound is delivered orally as a tablet.

[0025] In some embodiments, the TLR4 agonist selectively stimulates the TIR domain clustering adaptor-inducing interferon-β (TRIF) pathway.

[0026] In some embodiments, the TLR4 agonist reduces 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 a subject.

[0027] The present invention also provides a pharmaceutical composition for treating or preventing organ dysfunction in a subject in need thereof, the pharmaceutical composition comprising a colloidal preparation of a monophosphoryl lipid A (MPLA)-like compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the MPLA-like compound is a 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.

[0028] In some embodiments, the pharmaceutical composition is an aqueous composition. In other embodiments, the pharmaceutical composition is a dry powder.

[0029] In some embodiments, the pharmaceutical composition has a concentration of MPLA from about 1 μg / mL to about 10000 μg / mL.

[0030] In some embodiments, the composition further comprises a stabilizer, preferably, the stabilizer is trehalose.

[0031] In some embodiments, the composition comprises micelles having an average diameter or length of about 1 nm to about 1000 nm.

[0032] In some embodiments, the composition includes a bulking agent selected from one or more of the following: mannitol, trehalose, chitosan, HP-B-cyclodextrin, hydroxypropyl methylcellulose (HPMC), dextran, pea starch, and sucrose.

[0033] BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1:A. Stimulation of Toll-like receptor 4 (TLR4) can activate one of the following two pathways: 1) the myeloid differentiation primary response 88 (MyD88) pathway (which leads to the production of pro-inflammatory cytokines), and 2) the TIR domain clustering adaptor-induced interferon-b (TRIF) pathway (which leads to the production of anti-inflammatory protective cytokines); B. Monophosphoryl lipid A (MPLA) and MPLA-like compounds can preferentially stimulate TLR4 to activate the TRIF pathway, thereby leading to the production of protective cytokines.

[0035] Figure 2 : IP-10 upregulation as a function of dose using representative formulated PHADs. Based on an activity assay using mouse macrophages of the J774 cell line, various formulations of PHADs were evaluated in vitro in cells, measured by upregulation of IP-10 to demonstrate TLR4 stimulation in response to PHADs.

[0036] Figure 3 : Dose-dependent reduction in fibrosis (collagen fractional volume) as a function of total collagen deposition measured via PicoSirius Red in response to daily treatment with PHAD (REVTx-300), an MPLA-like compound, in the rat UUO model of acute kidney injury and chronic kidney disease.

[0037] Figure 4 : Concentration of TGF-β in rat serum at day 7 after UUO procedure in response to daily treatment with PHAD (REVTx-300), a MPLA-like compound, in a rat model of acute kidney injury and chronic kidney disease.

[0038] Figure 5 : Concentration of NGAL in rat serum on day 7 after UUO procedure in response to daily treatment with PHAD (REVTx-300), a MPLA-like compound, in a rat model of acute kidney injury and chronic kidney disease.

[0039] Figure 6 : Concentration of hepcidin in rat serum on day 7 after UUO procedure in response to daily treatment with PHAD (REVTx-300), a MPLA-like compound, in a rat model of acute kidney injury and chronic kidney disease.

[0040] Figure 7 : Concentration of heme oxygenase-1 in rat serum on day 7 after UUO procedure in response to daily treatment with PHAD (REVTx-300), a MPLA-like compound, in a rat model of acute kidney injury and chronic kidney disease.

[0041] Figure 8: Concentration of IL-10 in rat serum on day 7 after UUO procedure in response to daily treatment with PHAD (REVTx-300), a MPLA-like compound, in a rat model of acute kidney injury and chronic kidney disease.

[0042] Fig. 9 : Concentration of IL-1β in rat serum on day 7 after UUO procedure in response to daily treatment with PHAD (REVTx-300), a MPLA-like compound, in a rat model of acute kidney injury and chronic kidney disease.

[0043] Fig.10 : Concentration of IL-18 in rat serum on day 7 after UUO procedure in response to daily treatment with PHAD (REVTx-300), a MPLA-like compound, in a rat model of acute kidney injury and chronic kidney disease.

[0044] Detailed description

[0045] definition

[0046] "About" and "approximately" generally mean that the amount measured is within an acceptable degree of error, given the nature or precision of the measurement. Typically, exemplary degrees of error are within 20 percent (%) of a given value or range of values, preferably within 10%, and more preferably within 5%. Alternatively, and particularly in biological systems, the terms "about" and "approximately" can mean values ​​within an order of magnitude of a given value, preferably within 5 times of a given value, and more preferably within 2 times of a given value.

[0047] As used herein, "colloid" refers to any liquid or solid composition including a multi-molecular aggregate microstructure having a diameter or length in the range of 1 nm to 10 um. Such microstructures include (but are not limited to) micelles, liposomes, vesicles, nanoparticles, microparticles, etc. The microstructure can be spherical, elliptical, rectangular, flat or any other shape.

[0048] As used herein, "micelles" is an art-recognized term and refers to particles of colloidal size that exist in equilibrium with the molecules or ions in the solution in which they are formed. They are aggregates (or supramolecular assemblies) of molecules dispersed in a liquid, forming a colloidal suspension (also referred to as a related colloidal system). Typically micelles in water form aggregates in which the hydrophilic "head" region is in contact with the surrounding solvent, thereby isolating the hydrophobic single tail region in the center of the micelle.

[0049] As used herein, "liposome" is an art-recognized term and refers to a spherical vesicle having at least one lipid bilayer.Liposomes can be prepared by disrupting biological membranes, such as by sonication.

[0050] As used herein, "vesicle" is an art-recognized term and refers to a membranous fluid-filled sac surrounded by a lipid bilayer.

[0051] As used herein, "nanoparticle" is an art-recognized term and is generally defined as a particle of a substance having a diameter between 1 nanometer (nm) and 100 nanometers (nm). The term is sometimes used for larger particles, up to 500 nm.

[0052] As used herein, "microparticles" is an art-recognized term and is defined as particles having a size between 1 μm and 1000 μm.

[0053] The term "treatment" includes prophylactic treatment and / or therapeutic treatment. The term "prophylactic or therapeutic" treatment is recognized in the art and includes administering one or more of the subject compositions to the host. If administered before clinical manifestation of a harmful condition (e.g., a disease or other harmful state of the host animal), the treatment is prophylactic (i.e., it protects the host from developing a harmful condition), and if administered after manifestation of a harmful condition, the treatment is therapeutic (i.e., it is intended to reduce, alleviate or stabilize an existing harmful condition or its side effects). Treatment of respiratory viral infections may include: alleviating or eliminating symptoms (e.g., runny nose, sneezing, itchy and watery eyes, coughing, fatigue, headache, sore throat or congestion).

[0054] 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 relative to an untreated control sample, or a compound that delays the onset of or reduces the severity of one or more symptoms of a disorder or condition relative to an untreated control sample.

[0055] "Patient", "subject" or "individual" are used interchangeably and refer to humans or non-human animals. These terms include mammals (such as humans, non-human primates, livestock animals (including cattle, pigs, etc.)), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats). In some embodiments, the subject is a human.

[0056] As used herein, the phrase "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, composition or vehicle that one skilled in the art would consider suitable for making a pharmaceutical formulation suitable for administration to a subject, such as a liquid or solid filler, diluent, lubricant, binder, carrier, wetting agent, disintegrant, solvent or encapsulating material. Each excipient must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and is "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); cellulose and its derivatives (such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate); powdered tragacanth; malt; gelatin; talc; silica, waxes; oils (such as corn oil and sesame oil); glycols (such as propylene glycol and glycerol); polyols (such as sorbitol, mannitol, and polyethylene glycol); esters (such as ethyl oleate and ethyl laurate); agar; buffers; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; and other nontoxic compatible substances routinely used in pharmaceutical formulations.

[0057] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or medicament is an amount of the drug or medicament that, when administered to a subject, will have the desired therapeutic effect. The full therapeutic effect does not necessarily occur with the administration of a single dose, and may occur only after a series of doses are administered. Thus, a therapeutically effective amount may be administered in one or more administrations. The exact effective dose required for a subject will depend, for example, on the subject's size, health, and age, and the nature and extent of the condition being treated.

[0058] Activation of the MyD88 signaling pathway via TLR4 stimulation results in the production of pro-inflammatory cytokines (including IL-1β, IL-6, TNF-α, and IL-18) (Edilova). Long-term exposure to these cytokines has been associated with inflammatory processes that lead to tissue damage in chronic diseases. MPLA-like compounds primarily signal through the TRIF pathway via TLR4 stimulation, thereby reducing the production of pro-inflammatory cytokines and producing anti-inflammatory cytokines (such as IFN-β, IL-4, IL-10, IP-10, and TGF-β) to correct this dysregulation, thereby slowing or inhibiting the progression of chronic organ diseases. TLR4-mediated TRIF bias supports the potential correction of cellular activities associated with certain conditions, which are associated with cytokine dysregulation, including inflammatory diseases, possible macrophage activation syndrome (which promotes the cytokine storm observed in sepsis and ARDS), and the recently observed "inflammatory aging" (which has been used to describe the increase in inflammation observed with increasing age) (Rea, 2018).

[0059] The inflammatory process is driven by the cell activity initiated in response to pro-inflammatory cytokines. In the complex cascade of the inflammatory process in response to tissue or cell damage, a series of cell communications are initiated. At the initial injury site, sentinel dendritic cells (DC) are activated, which cause the generation of multiple cytokines (including IL-12 and IL-18), which in turn activate natural killer cells (NK cells). These activated NK cells can then release a large amount of highly cytotoxic IFN-γ, ultimately leading to cell death (Zwirner). Chronic DC and NK cell activation induces the production of pro-inflammatory cytokines, which lead to a cytotoxic environment, thereby promoting an inflammatory state.

[0060] While not being bound by theory, MPLA is not thought to completely disrupt the production of pro-inflammatory cytokines; rather, MPLA enhances host resistance during inflammatory events by attenuating the production of pro-inflammatory cytokines, which enables MPLA to fight infection while reducing tissue and organ damage (Watts). The reduced inflammatory signaling from TLR4 binding to MPLA, along with minimal impairment of the immunostimulatory adjuvant effect on T cells (Thompson et al., 2005; Mata-Haro et al., 2007) suggests that MPLA may prove to be safe and effective for use as a monotherapy for certain chronic inflammatory conditions.

[0061] A representative example of a MPLA-like compound (and also a representative example of the major species of MPLA derived from bacteria), the structure of a synthetic phosphorylated hexaacyl disaccharide (PHAD) is shown below:

[0062]

[0063] The common feature of all MPLA-like compounds is a monophosphorylated disaccharide. The degree of acylation can vary from as few as 4 acyl groups to as many as 9 acyl groups. In addition, the length of each acyl chain (e.g., the number of carbons) can vary from about 8 to about 20 carbons.

[0064] MPLA-like compounds can be synthetic or biologically derived as described (e.g., derived from 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.

[0065] In some embodiments of the invention, organ function loss associated with chronic organ disease can be treated by administering an effective amount of a TLR4 agonist to a subject. In certain embodiments, the TLR4 agonist is a class MPLA compound. In certain preferred embodiments, the class MPLA compound is synthetic and is selected from 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. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0066] In some preferred embodiments, the TLR4 agonist selectively stimulates the TRIF pathway over the myeloid differentiation primary response 88 (MyD88) pathway.

[0067] Chronic Disease and the Innate Immune System

[0068] Chronic organ diseases are characterized by progressive organ function loss. There are many chronic diseases of important organs, including chronic kidney disease (CKD), non-alcoholic steatohepatitis (NASH), osteoarthritis (OA), rheumatoid arthritis (RA), irritable bowel syndrome (IBS), pulmonary fibrosis, heart disease, etc. Many of these chronic diseases are the result of single stress or repeated stress caused by internal or external stimuli.

[0069] Chronic disease of organs (due to chronic inflammation and subsequent fibrosis) follows the following pattern: permanent and ongoing destruction of living, functional cells and subsequent replacement by non-functional proteins, collagen, resulting in fibrosis (scar tissue). The establishment of fibrosis and the subsequent death of organs is driven by a persistent inflammatory process associated with an innate immune response. Redirecting the innate immune response from a pro-inflammatory state to an anti-inflammatory (non-inflammatory or protective) state may slow or stop the progressive destruction and scarring of organ tissue, allowing the healing process to occur.

[0070] The present invention contemplates the use of 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 via TLR4 stimulation results in the production of pro-inflammatory cytokines (including IL-1β, IL-6, TNF-α, and IL-18) (Edilova). Long-term exposure to these cytokines has been associated with inflammatory processes that cause tissue damage in chronic diseases. MPLA-like compounds primarily signal through the TRIF pathway via TLR4 stimulation, thereby reducing the production of pro-inflammatory cytokines, and producing anti-inflammatory cytokines (such as IFN-β, IL-4, IL-10, IP-10, and TGF-β) to correct the dysregulation, thereby slowing or inhibiting the progression of chronic organ diseases.

[0071] TLR4-mediated TRIF bias supports potential correction of cellular activities associated with certain conditions that are associated with cytokine dysregulation, including inflammatory diseases, possible macrophage activation syndrome (which contributes to the cytokine storm observed in sepsis and ARDS), and the recently observed "inflammaging" (which has been used to describe the increase in inflammation observed with increasing age) (Rea, 2018).

[0072] The inflammatory process is driven by the cell activity initiated in response to pro-inflammatory cytokines. In the complex cascade of the inflammatory process in response to tissue or cell damage, a series of cell communications are initiated. At the initial injury site, sentinel dendritic cells (DC) are activated, which cause the generation of multiple cytokines (including IL-12 and IL-18), which in turn activate natural killer cells (NK cells). These activated NK cells can then release a large amount of highly cytotoxic IFN-γ, ultimately leading to cell death (Zwirner). Chronic DC and NK cell activation induces the production of pro-inflammatory cytokines, which lead to a cytotoxic environment, thereby contributing to the inflammatory state.

[0073] The efficient production of pro-inflammatory cytokines depends on the assembly of inflammasomes. Inflammasomes are multiprotein structures formed in the cytoplasm of activated innate immune cells, which lead to the maturation of IL-1β and IL-18 from inactive precursor proteins to their active, mature forms (Chilton). Once the inflammasome has been established, the production of pro-inflammatory cytokines becomes possible via the MyD88 pathway.

[0074] The relatively low levels of MyD88-driven activity observed as a result of MPLA-mediated TLR4 activation are likely due to a failure in inflammasome assembly (Embry et al., 2011). MPLA produces reduced levels of IL-1β, IL-6, and TNF-α relative to LPS-mediated responses (Guo, Chentouh, Watts). These reduced levels of pro-inflammatory cytokines may result in insufficient signals required to activate DCs, which prevents the production of IL-12 and IL-18 (signals required to activate NK cells), thereby closing the feedback loop of IFN-γ-mediated tissue damage associated with inflammatory diseases.

[0075] In some embodiments, the TLR4 agonists described herein selectively activate the TRIF pathway over the MyD88 pathway. In certain embodiments, the TLR4 agonists have ECs for activation of the TRIF pathway. 50 , and it targets ECs that activate the MyD88 pathway 50 Having a ratio of about 1.1:1 to greater than 100,000:1. In some embodiments, these EC 50 In some embodiments, the ratio of these ECs 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 5000:1, greater than about 10000:1, or greater than about 100000:1. 50 The ratio is in the range of about 1:1 to about 100000:1, about 1.1:1 to about 50000:1, about 1.1:1 to about 10000:1, about 1.1:1 to about 1000:1, 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.

[0076] Inflammation is a necessary process in response to damage. Inflammation allows the infiltration of macrophages, T cells and B cells to help counter damage or infection, pathogens and the cell excavation (excavation) of cell fragments as a result of tissue damage, and the final regression of damage or infection. MPLA does not completely stop the production of proinflammatory cytokines; on the contrary, MPLA enhances host resistance during inflammatory events by the weakening of the production of proinflammatory cytokines, which enables MPLA to counter infection while alleviating tissue and organ damage (Watts). For example, when compared with those IL-12 produced by untreated DC, the low amount of IL-12 produced by the DC processed by MPLA is very important. Although these low amounts of IL-12 cannot fully activate NK cells, these low amounts of IL-12 are enough to induce the effective activation of T cells (Ismaili). The reduced inflammatory signaling from TLR4 binding to MPLA, together with minimal impairment of the immunostimulatory adjuvant effect on the initial clonal expansion of T cells (Thomson et al., 2005; Mata-Haro et al., 2007), suggests that MPLA may prove to be safe and effective for use as a monotherapy for certain chronic inflammatory conditions.

[0077] In some embodiments, organ function loss associated with chronic organ diseases attributed to inflammation and fibrosis can be treated by administering an effective amount of a TLR4 agonist to a subject. In some embodiments, the TLR4 agonist is a class MPLA compound. In some preferred embodiments, the class MPLA compound is synthetic and selected from phosphorylated hexaacyl disaccharides (PHAD), 3-deacylated phosphorylated hexaacyl disaccharides (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharides (3D(6-acyl) PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0078] Monophosphoryl lipid A (MPLA)-like compounds can selectively stimulate TLR4 to activate the TRIF pathway, leading to 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.

[0079] In other embodiments, long-term organ dysfunction (long-term organ dysfunction is the result of acute inflammation due to internal or external stimulation) can be treated by administering an effective amount of a TLR4 agonist to the subject. In some embodiments, the TLR4 agonist is a class MPLA compound. In some preferred embodiments, MPLA is synthetic and is selected from 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. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0080] In some preferred embodiments, the source of acute inflammation comprises one or more of the following non-limiting examples: toxicity from drugs, toxicity from chemotherapy, ischemia, trauma, cancer, and infection.

[0081] Chronic Kidney Disease (CKD)

[0082] Kidney disease is a major public health problem, affecting ~10% of the population in industrialized countries (1). Acute kidney injury (AKI), which affects 13.3 million people each year, can lead to chronic kidney disease (CKD). Both acute kidney injury (AKI) and chronic kidney disease (CKD) are increasing worldwide (2). Progression of chronic kidney damage often leads to end-stage renal disease, requiring renal replacement therapy (dialysis or transplantation), resulting in significant morbidity and mortality in affected patients.

[0083] CKD can be initiated and propagated in several ways. One common condition is the high blood sugar levels associated with diabetes (type 1 or type 2). High blood sugar is toxic to kidney cells, causing stress that mimics the inflammatory process, leading to the death of these cells and subsequent fibrosis, ultimately causing a sustained loss of kidney function over time. High arterial blood pressure is another stressor that triggers the inflammatory process that leads to CKD.

[0084] Other causes of CKD include: glomerulonephritis (inflammation in the glomeruli), polycystic kidney disease, autoimmune diseases (such as systemic lupus erythematosus), vesicoureteral reflux (a condition in which urine flows back into the kidneys), pyelonephritis, interstitial nephritis (inflammation of the kidney tubules), kidney stones, kidney obstruction or cancer which can cause the kidneys to fail over time, overuse of certain medications, drug abuse (heroin or cocaine), chemotherapy (such as cisplatin).

[0085] 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 measurement of true glomerular filtration rate, measurement of blood creatinine, measurement of blood urea nitrogen, measurement of urine albumin, determination of the urine albumin to creatinine ratio.

[0086] 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 an effective amount of a TLR4 agonist to a subject. 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 disaccharides (PHAD), 3-deacylated phosphorylated hexaacyl disaccharides (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharides (3D(6-acyl) PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0087] In other embodiments, organ dysfunction (organ dysfunction is a result of acute inflammation due to acute internal or external stress) can be prevented by administering an effective amount of a TLR4 agonist to the subject. In some preferred embodiments, the TLR4 agonist is an MPLA compound. In other preferred embodiments, MPLA is synthetic and selected from phosphorylated hexaacyl disaccharides (PHAD), 3-deacylated phosphorylated hexaacyl disaccharides (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharides (3D(6-acyl) PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0088] In some preferred embodiments, acute stress is an ischemic event, such as during renal surgery or during a hypotensive state, such as during severe infection (sepsis). Ischemic acute kidney injury (AKI) causes ATP depletion, which causes changes in epithelial and endothelial cells. The cell changes cause destruction, thereby reducing the glomerular filtration rate. Cell death caused as a result of AKI is mediated by apoptosis and necrosis. Complex cell interactions use certain inflammatory mediators that promote sustained damage during active tubular necrosis to guide the activity between damage or repair (Sharfuddin). In other preferred embodiments, acute stress is attributed to toxicity as a result of drug overdose or substance abuse. In other preferred embodiments, stress is attributed to toxicity, and toxicity is attributed to chemotherapy. In a preferred embodiment, stress is induced by platinum-containing chemotherapy (such as cisplatin). In another preferred embodiment, the acute stress that causes AKI is attributed to cardiac surgery.

[0089] In the surgical setting, ischemia may be intentionally induced, such as during a surgical procedure requiring cardiopulmonary bypass. Ischemia may be the unintended consequence of adverse complications such as intraoperative hypotension. Regardless of the cause, an ischemic event initially deprives the affected area(s) of blood, oxygen, and other nutrients, and subsequently can exacerbate the injury when blood supply returns to the site along with reactive oxygen species (ROS) and other components that cause oxidative stress to the tissue.

[0090] Ischemic preconditioning and the resulting ischemic tolerance of various organs (e.g., brain, heart, liver, intestine, skeletal muscle) is an adaptive defense mechanism in 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 damage upon exposure to subsequent ischemic events by reducing excitotoxicity, apoptosis, and inflammation, thereby protecting mitochondria and enhancing antioxidant mechanisms (Bhuiyan 2010). Nitrogen interventions that promote ischemic tolerance in the absence of oxygen deprivation techniques are a safer perioperative approach to ischemic preconditioning.

[0091] Nonalcoholic steatohepatitis (NASH)

[0092] Non-alcoholic fatty liver disease (NAFLD) is a condition in which excess fat accumulates in the liver. This fat accumulation is not caused by heavy drinking. When heavy drinking causes fat to accumulate in the liver, the condition is called alcohol-related liver disease.

[0093] The two types of NAFLD are nonalcoholic fatty liver disease (NAFL) and nonalcoholic steatohepatitis (NASH). People usually develop one type of NAFLD or the other, although sometimes people with one form are later diagnosed with the other.

[0094] NASH is a form of NAFLD in which, in addition to fat in the liver, patients also have liver inflammation and liver damage. The inflammation and liver damage of NASH can lead to fibrosis, or scarring, of the liver. NASH can lead to cirrhosis, in which the liver becomes scarred and permanently damaged. Cirrhosis can lead to liver cancer.

[0095] In some preferred embodiments of the present invention, the loss of liver function associated with the progression of NASH is prevented by administering an effective amount of a TLR4 agonist to a subject. In some preferred embodiments, the TLR4 agonist MPLA compound. In a more preferred embodiment, the MPLA compound is synthetic and is selected from 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. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0096] Osteoarthritis

[0097] Osteoarthritis is the most common form of arthritis in the elderly, and it is one of the most common causes of physical disability in the elderly. The 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 cartilage (the tissue that cushions the ends of bones in the joints) breaks down and wears away. The inflammatory process drives the destruction of cartilage and joints. In extreme cases, all the cartilage is worn away, exposing the bones to rub directly against each other, and if possible, an artificial joint needs to be inserted.

[0098] In some preferred embodiments of the present invention, joint function loss associated with OA progression is prevented by administering an effective amount of a TLR4 agonist to a subject. 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 disaccharides (PHAD), 3-deacylated phosphorylated hexaacyl disaccharides (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharides (3D(6-acyl) PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0099] Rheumatoid arthritis (RA)

[0100] RA is an autoimmune and inflammatory disease that primarily attacks the joints, often multiple joints at a time. RA commonly affects the joints of the hands, wrists, and knees. In joints with RA, the lining of the joint becomes inflamed, causing damage to the joint tissue. This tissue damage can cause long-term or chronic pain, instability (lack of balance), and deformity (deformity). RA can also affect other tissues throughout the body and cause problems with organs such as the lungs, heart, and eyes.

[0101] In some preferred embodiments of the present invention, the joint function loss associated with RA progression is prevented by administering an effective amount of a TLR4 agonist to a subject. 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 disaccharides (PHAD), 3-deacylated phosphorylated hexaacyl disaccharides (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharides (3D(6-acyl) PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0102] Irritable bowel syndrome

[0103] Irritable bowel syndrome (IBS) is a chronic disorder that affects the gastrointestinal tract, causing abdominal pain, bloating, cramping, flatulence, diarrhea, constipation, or both. Although common, only a small percentage of patients with IBS have severe symptoms in conditions such as Crohn’s disease and ulcerative colitis, in which persistent mucosal inflammation has been observed at the microscopic and molecular levels. In some cases, patients may develop post-infectious IBS as a result of infectious gastroenteritis, which can lead to systemic inflammation, perpetuating a cycle of chronic, low-grade, subclinical inflammation. Upregulated IL-1β has been observed in rectal biopsies of patients with post-infectious IBS. Correction or normalization of cytokine secretion may not only reduce the inflammation that causes IBS, but also help reestablish a healthy gut flora population.

[0104] In some preferred embodiments of the present invention, the reduction of inflammation that may promote the progression of IBS or inflammatory bowel disease is prevented by administering an effective amount of a TLR4 agonist to a subject. In some preferred embodiments, the TLR4 agonist is a class MPLA compound. In a more preferred embodiment, the class MPLA compound is synthetic and is selected from phosphorylated hexaacyl disaccharides (PHAD), 3-deacylated phosphorylated hexaacyl disaccharides (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharides (3D(6-acyl) PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0105] Pulmonary fibrosis

[0106] Fibrotic lung disease, including idiopathic pulmonary fibrosis, is a broad term that describes inflammation and scarring (fibrosis) of lung tissue that causes reduced lung function. Fibrotic lung disease can be caused by a variety of factors, including long-term exposure to toxins and pollutants, radiation and radiation therapy, and certain medications, such as chemotherapy drugs. Drugs designed to kill cancer cells, such as methotrexate (Trexall, Otrexup, and others) and cyclophosphamide, can also damage lung tissue. Some drugs used to treat irregular heartbeats, such as amiodarone (Cordarone, Nexterone, Pacerone), can damage lung tissue. Antibiotics, such as nitrofurantoin (Macrobid, Macrodantin, and others) or ethambutol, can cause lung damage. Certain anti-inflammatory drugs, such as rituximab (Rituxan) or sulfasalazine (Azulfidine), can cause lung damage. Certain underlying conditions (eg, dermatomyositis, polymyositis, mixed connective tissue disease, systemic lupus erythematosus, rheumatoid arthritis, sarcoidosis, scleroderma, pneumonia) may benefit from more effective anti-inflammatory therapy.

[0107] In some preferred embodiments of the present invention, the loss of lung function associated with the progression of pulmonary fibrosis is prevented by administering an effective amount of a TLR4 agonist to a subject. 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 disaccharides (PHAD), 3-deacylated phosphorylated hexaacyl disaccharides (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharides (3D(6-acyl) PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0108] heart disease

[0109] Cardiovascular disease (CVD) is a group of diseases that affect the heart or cardiovascular system, accounting for 31% of all deaths and remains the leading cause of mortality worldwide. Ischemic heart disease and endocardial fibrosis are the main causes of end-stage heart failure.

[0110] Fibrosis is a major cause of morbidity and mortality from cardiac disease. Fibrotic scarring of the cardiac muscle most commonly occurs after myocardial infarction; however, a variety of other conditions promote cardiac fibrosis, such as hypertensive heart disease, diabetic hypertrophic cardiomyopathy, and idiopathic dilated cardiomyopathy [4,5].

[0111] In some preferred embodiments of the present invention, the loss of cardiac function associated with cardiac fibrosis is prevented by administering an effective amount of a TLR4 agonist to a subject. 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 disaccharides (PHAD), 3-deacylated phosphorylated hexaacyl disaccharides (3D-PHAD), 3D-(6-acyl) phosphorylated hexaacyl disaccharides (3D(6-acyl) PHAD) or a pharmaceutically acceptable salt thereof. In other embodiments, one or more synthetic MPLAs are co-administered together.

[0112] MPLA-like compounds

[0113] The pharmaceutical compositions of the present disclosure include monophosphoryl lipid A (MPLA)-like compounds. MPLA was originally isolated from lipopolysaccharides obtained from the cell walls of Gram-negative bacteria:

[0114]

[0115] MPLA derived from bacteria is generally a mixture of several different species. One of the main species of MPLA derived from bacteria is shown. As an example, MPLA can be derived from Salmonella Minnesota R595 lipopolysaccharide. As will be understood, MPLA may also be derived from other Salmonella species. Bacterial LPS can be treated via continuous acid and alkaline hydrolysis steps to remove polysaccharide side chains, phosphate groups, and partially remove acetyl side groups. Crude MPLA can then be purified. The final MPLA product is a mixture of β1à6 connected by seven acyl-, six acyl- and five acyl-monophosphate glucosamine disaccharides. Diacetyl, triacetyl and tetraacetyl, if present, are considered impurities. Acylated lipids vary and include lauroyl, myristoyl and palmitoyl. Although the relative proportions of each species may vary from batch to batch, the main species produced is a six-acylated disaccharide product.

[0116] The major species found in bacterially derived MPLA have been chemically synthesized and have immunostimulatory properties comparable to those of the bacterially derived species. Examples of synthetic MPLA compounds suitable for use in the present invention include phosphorylated hexaacyl disaccharides (also known as glucopyranosyl lipid A or GLA), 3D-PHAD (or 3-acyl-PHAD) (also known as monophosphoryl 3-deacyl lipid A):

[0117]

[0118] and 3D-(6-acyl)PHAD (or 3,6-acylPHAD):

[0119]

[0120] Synthetic variants of MPLA that are also suitable and within the scope of the present invention include those in which the fatty acid chain length varies between 10-20 carbons, and those in which the degree of acylation is penta-, hexa-, or hepta-.

[0121] PHAD is chemically equivalent to the main component of MPLA derived from bacteria. PHAD is also biologically equivalent to MPLA derived from bacteria.

[0122] Dosage Form and Route of Administration

[0123] The MPLA-like compounds can be administered by several different routes. The choice of route depends on a variety of factors, including the need (or lack thereof) for systemic exposure, the desire for the MPLA-like compound to reach a specific organ quickly, and patient tolerance and compliance.

[0124] Methods of systemic delivery include those methods known in the art that provide active molecules (e.g., drugs) to be distributed throughout the body's circulatory system. Systemic delivery methods include intramuscular, intravenous, subcutaneous, intraperitoneal, sublingual, and oral. As will be appreciated, any systemic delivery method is suitable for use with the present invention. Particularly suitable systemic delivery methods include oral delivery, intramuscular delivery, and intravenous delivery.

[0125] In some embodiments, it may be desirable to have the drug interact only with mucosal tissues, and there is no systemic exposure or minimal systemic exposure there. Methods for mucosal delivery include those methods known in the art for providing active molecules to the delivery of mucosa. Mucosal delivery methods include intranasal, intrabuccal, sublingual and oral. Particularly suitable methods for mucosal delivery include intranasal delivery.

[0126] In these embodiments, the composition comprising the MPLA compound can be formulated for delivery to the nasal passage or nasal vestibule of a subject as drops, aerosols, micelles in solution, lipid or liquid nanospheres, liposomes, lipid or liquid microspheres, solution sprays, or powders. The composition can be administered by direct application to the nasal passage or can be atomized or made into a spray for inhalation through the nose or mouth.

[0127] In some embodiments, the method comprises applying a nasal spray, a medicated nasal swab, a medicated wipe, nasal drops, or an aerosol to the nasal passage or nasal vestibule of the subject. To this end, viscosity modifiers that can be employed to optimize the product for the application form can include cetyl alcohol, stearyl alcohol, carnauba wax, stearic acid, xanthan gum, magnesium aluminum silicate, gelatin, carbomer, poloxamer, PEG, waxes, starches, castor oil derivatives, fatty acids, fatty alcohols, and lecithin.

[0128] In some embodiments, a small needle-free nasal spray device (which can allow (self) administration with little or no prior training to deliver the desired dose) can be used to deliver the composition of the present invention. The device may include a reservoir containing a certain number of compositions. The device may include a pump spray for delivering one or more metered doses to the nasal cavity of the subject. The device can advantageously be used in single doses or multiple doses. It can be further designed to administer the expected dose with multiple sprays (e.g., two sprays, such as once per nostril, or as a single spray, such as in one nostril), or designed to change the dose according to the patient's weight or maturity. In some embodiments, nasal drops can be prepackaged in a storage bag or ampoule that can be opened immediately before use and squeezed or sprayed into the nasal passage. In some embodiments, the product powder can be reconstituted with an aqueous solvent immediately before administration to complete the nasal spray or drops when used. In some embodiments, nasal sprays or drops may be accomplished by reconstituting a solid drug product powder contained in a suitable delivery device with an aqueous vehicle for some period of time prior to patient administration during which the drug product is considered stable in solution.

[0129] In certain embodiments, the composition is suitable for parenteral administration to a mammal, most preferably 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, etc. Through the appropriate use of excipients and packaging and delivery means known in the art, the composition may be suitable for direct injection or intravenous infusion, or for addition to an intravenous drip for gradual infusion.

[0130] In other embodiments, the present invention provides a pharmaceutical package comprising a vial or ampoule containing a 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, a written and / or pictorial description of the active ingredient, instructions for diluting the composition to a concentration suitable for administration, applicable indications, suitable dosing regimens, contraindications, drug interactions, and any adverse side effects noted during clinical trials.

[0131] In an alternative embodiment, the pharmaceutical pack may comprise a plastic bag containing 100 ml to 2 L of the pharmaceutical composition of the invention in the form of a solution suitable for intravenous administration, together with instructions for use as described above.

[0132] In alternative embodiments, the pharmaceutical compositions of the invention may be in a form suitable for oral administration (such as, for example, a syrup or palatable solution); a form suitable for topical application (such as, for example, a cream or 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 compositions for alternative routes of administration are well known in the art, and it is expected that those skilled in the relevant art can adapt these known methods to the MPLA-like compounds and formulations described in the present invention.

[0133] The present invention provides pharmaceutically acceptable compositions, including a therapeutically effective amount of one or more MPLA compounds formulated with one or more pharmaceutically acceptable excipients. The pharmaceutical compositions 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), administration by inhalation (e.g., aerosols, solutions for atomization or dry powders); parenteral administration (e.g., sterile solutions or suspensions); topical application (e.g., emulsions, creams, ointments or sprays); intraocular administration; or vaginal or rectal administration (e.g., vaginal suppositories, suppositories, creams or foams). Preferably, the pharmaceutical formulation is suitable for parenteral administration, and more preferably, it is a non-pyrogenic solution suitable for intravenous administration.

[0134] Tablets can be made by compression or molding with one or more auxiliary ingredients, optionally. Compressed tablets can be prepared by using a binder (e.g., gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), a surfactant or a dispersant. Molded tablets can be made by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0135] Tablets and other solid dosage forms of the pharmaceutical compositions of the present invention may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings known in the art of pharmaceutical preparations. Hydroxypropylmethylcellulose, other polymer matrices, liposomes and / or microspheres in various proportions to provide the desired release profile may also be used, for example, to prepare them so as to provide a slow or controlled release of the modified substance therein. They may be sterilized by, for example, filtering through a bacteria-retaining filter, or by incorporating a disinfectant into the form of a sterile solid composition (which may be dissolved in sterile water or some other sterile injectable medium immediately before use). These compositions may also optionally contain an opacifying agent, and may be compositions of them that release the active ingredient(s) only or preferably in a specific portion of the gastrointestinal tract in a delayed manner. Examples of embedding compositions that may be used include polymeric substances and waxes. The MPLA-like compounds may also be in microencapsulated form with one or more of the excipients described above (if appropriate).

[0136] 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 and other solvents, solubilizers and emulsifiers.

[0137] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0138] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum oxide hydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0139] In dry powder formulations suitable for inhalation, the particle size of the particulate drug should be such that substantially all of the drug is inhaled into the lungs when the aerosol formulation is administered, and is therefore expected to be less than 20 microns, preferably in the range of 1 micron to 10 microns, more preferably 1 micron to 5 microns. The particle size of the drug can be reduced by conventional means, such as by grinding or micronization. The aerosol formulation preferably contains 0.5-30% w / w of the MPLA-like compound relative to the total weight of the formulation.

[0140] The propellant may optionally contain an adjuvant having a higher polarity and / or a higher boiling point than the propellant. Polar adjuvants that may be used include (e.g., C 2-6) aliphatic alcohols and polyols, such as ethanol, isopropanol and propylene glycol, preferably ethanol. Typically, only a small amount of polar adjuvant (e.g. 0.05-3.0% w / w) may be required to improve the stability of the dispersion. However, the formulation of the present invention is preferably substantially free of polar adjuvants, particularly 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). Typically, up to 50% w / w of the propellant may include a volatile adjuvant, such as 1% w / w to 30% w / w of a volatile saturated C1-C6 hydrocarbon.

[0141] Aerosol formulations according to the invention may optionally include one or more surfactants which are physiologically acceptable when administered by inhalation.

[0142] For administration by inhalation, gelatin, plastic or other capsules, cartridges, blister packs and / or strips are optionally used, and the medication is suitably inhaled from a nebulizer, from a pressurized metered dose inhaler or as a dry powder from a dry powder inhaler.

[0143] Administration of the drug may be indicated for the treatment of mild, moderate or severe acute or chronic symptoms, or for preventive treatment. It should be understood that the precise dose administered will depend on the age and condition of the patient, the specific granular drug used and the frequency of administration, and will ultimately be at the discretion of the attending physician. Typically, administration will vary from once a day to four times a day or more.

[0144] For use in dry powder inhalers, the active ingredient can be modified by spray drying or compression to form a powder with suitable flow characteristics. More commonly, a diluent or carrier that is usually nontoxic and inert is added to the drug. 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 a surface modifier, pretreatment to form low roughness particles, glidants and flavor masking agents or modifiers.

[0145] Pharmaceutical compositions of the present invention suitable for parenteral administration include a combination of an MPLA-like compound and one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions or sterile powders that can be reconstituted into sterile injectable solutions or dispersions before use, and the sterile powders may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0146] These compositions may also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants. Prevention of the action of microorganisms may be ensured by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, etc. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, etc., in the composition.

[0147] 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, α-tocopherol, and chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0148] The injectable depot form is made by forming a microencapsulation matrix of the title compound in a biodegradable polymer (such as polylactic acid-polyglycolic acid). According to the ratio of the drug to the polymer and the properties of the specific polymer adopted, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations are also prepared by embedding the drug in a liposome or microemulsion compatible with body tissues.

[0149] Dosage forms for topical or transdermal administration of the compounds of the invention include powders, sprays, ointments, pastes, creams, emulsions, gels, solutions, patches and inhalants. The MPLA-like compounds can be mixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers or propellants that may be required.

[0150] In addition to the active compounds of the invention, ointments, pastes, creams and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide or mixtures thereof. Ophthalmic preparations, eye ointments, powders, solutions and the like are also contemplated as being within the scope of the invention.

[0151] Formulations of the invention suitable for vaginal administration include vaginal suppositories, tampons, creams, gels, pastes, foams, or spray formulations containing suitable carriers known in the art. Such formulations can be prepared, for example, by mixing one or more MPLA 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 thus will melt in the rectum or vaginal cavity and release the MPLA compound).

[0152] Treatment dose and duration

[0153] In certain embodiments, the MPLA-like compound is administered at a total dose of between 0.001 mg and 100 mg, depending on the route of administration (e.g., parenteral, oral) and the therapeutic target (acute disease vs. chronic disease). In a preferred embodiment, the total dose administered is between 50 micrograms and 1000 micrograms. In a particularly preferred embodiment, the total dose is about 100 micrograms to 500 micrograms. In other preferred embodiments, the total dose is about 5-20 mg. In another preferred embodiment, the total dose is about 50-100 mg.

[0154] In other embodiments of the invention, the MPLA-like compound is administered in 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, biweekly, weekly, or monthly. The exact dosing frequency and the dose required for each interval will depend on a variety of factors, including the type of chronic organ disease being treated, the rate of disease progression, and patient tolerance. Early in the treatment phase, the dose and / or dosing frequency may be large, and as the progression of the disease slows, both the dose and / or dosing frequency are gradually reduced to maintain a stable state (lack of disease progression).

[0155] Compositions and methods of preparation

[0156] 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 comprise up to 15% of the total final volume of the drug product solution administered. In preferred embodiments, the organic solvent may comprise up to 5% of the total final volume of the drug product solution administered.

[0157] In certain embodiments, the organic solvent is miscible with water, such as an organic solvent selected from alcohol, glycerol, low molecular weight polyethylene glycol and low molecular weight poloxamer. In certain preferred embodiments, the organic solvent is an alcohol, such as methanol, ethanol, isopropanol, tert-butyl alcohol or preferably ethanol.

[0158] In certain embodiments, the pharmaceutical composition also includes one or more surfactants. In certain embodiments, one or more surfactants are selected from carboxymethyl cellulose, dodecyl trimethyl ammonium bromide (DTAB), n-dodecyl eight (ethylene oxide) (C12E8), n-dodecyl tetra (ethylene oxide) (C12E4), dioctanoyl phosphatidylcholine (C8-lecithin), polyethylene glycol (35) castor oil, polyoxyethylene castor oil EL (CrEL), octaethylene glycol monolauryl ether (C12E8), hexadecyl trimethyl ammonium bromide (CTAB), polypropylene oxide (PPO), polyethylene oxide (PEO), PEO-poly (D, L-lactic acid-to-caprolactone) (PEO-PDLLA) and sodium lauryl sulfate (SDS). In certain preferred embodiments, one or more surfactants are carboxymethyl cellulose.

[0159] In certain embodiments, the pharmaceutical composition may contain excipients, additives, bulking agents and mucoadhesives. These may include mannitol, trehalose, cyclodextrin and hydroxypropyl methylcellulose (HPMC). In a preferred embodiment, the excipients HP-β-cyclodextrin and trehalose are used.

[0160] In certain embodiments, the pharmaceutical composition further comprises a phospholipid selected from the group consisting of phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), sphingomyelin (including brain sphingomyelin), lecithin, lysolecithin, lysosphatidylethanolamine, cerebrosides, arachidylphosphatidylcholine (DAPC), didecanoyl-L-α-phosphatidylcholine (DDPC), di-elaidoylphosphatidylcholine (DEPC), dilauroylphosphatidylcholine (DLPC), dilinoleoylphosphatidylcholine, dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC) , distearoylphosphatidylcholine (DSPC), 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), arachidonylphosphatidylglycerol (DAPG), didecanoyl-L-α-phosphatidylglycerol (DDPG), di-elaidoylphosphatidylglycerol (DEPG), dilauroylphosphatidylglycerol (DLPG), dilinoleoylphosphatidylglycerol, dimyristoylphosphatidylglycerol (DMPG), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylglycerol (DSPG), 1-palmitoyl-2-oleoylphosphatidylglycerol (POPG), arachidonylphosphatidylethanolamine (DAPE), didecanoyl-L-α-phosphatidylethanolamine (DDPE), Di-elaidoylphosphatidylethanolamine (DEPE), dilauroylphosphatidylethanolamine (DLPE), dilinoleoylphosphatidylethanolamine, dimyristoylphosphatidylethanolamine (DMPE), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylethanolamine (DSPE), 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE), arachidonylphosphatidylinositol (DAPI), didecanoyl-L-α-phosphatidylinositol (DDPI), di-elaidoylphosphatidylethanolamine (DEPI), dilauroylphosphatidylinositol (DLPI), dilinoleoylphosphatidylethanolamine, dimyristoylphosphatidylethanolamine (DMPI), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylethanolamine (DSPE), 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE), arachidonylphosphatidylinositol (DAPI), didecanoyl-L-α-phosphatidylinositol (DDPI), di-elaidoylphosphatidylinositol (DEPI), dilauroylphosphatidylinositol (DLPI), dilinoleoylphosphatidylinositol, dimyristoylphosphatidylinositol (DMPI), dioleoylphosphatidylethanolamine ( DOPI), dipalmitoylphosphatidylinositol (DPPI), distearoylphosphatidylinositol (DSPI), 1-palmitoyl-2-oleoylphosphatidylinositol (POPI), arachidonylphosphatidylserine (DAPS), didecanoyl-L-α-phosphatidylserine (DDPS), di-antioleylphosphatidylserine (DEPS), dilauroylphosphatidylserine (DLPS), dilinoleoylphosphatidylserine, dimyristoylphosphatidylserine (DMPS), dioleoylphosphatidylserine (DOPS), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylserine (DSPS), 1-palmitoyl-2-oleoylphosphatidylserine (POPS), arachidonyl sphingomyelin,Didecanoyl sphingomyelin, di-antioleyl sphingomyelin, dilauroyl sphingomyelin, dilinoleoyl sphingomyelin, dimyristoyl sphingomyelin, sphingomyelin, dioleoyl sphingomyelin, dipalmitoyl sphingomyelin, distearoyl sphingomyelin, 1-palmitoyl-2-oleoyl sphingomyelin, and any combination thereof.

[0161] In certain embodiments, the pharmaceutical composition is free of salt (eg, NaCl) or substantially free of salt (eg, NaCl).

[0162] In certain embodiments, the pharmaceutical composition has an MPLA concentration of about 1 μg / mL to about 10000 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 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.

[0163] In certain embodiments, the pharmaceutical composition further comprises a stabilizer or multiple stabilizers. In certain preferred embodiments, the stabilizer is trehalose. In other preferred embodiments, the stabilizer is HP-β-cyclodextrin. In some most preferred embodiments, both HP-β-cyclodextrin and trehalose are used as stabilizers.

[0164] In certain embodiments, the pharmaceutical composition is a dry powder.

[0165] In certain embodiments, the colloid or micellar solution includes particles with an average diameter of about 1nm to about 1000nm. In certain preferred embodiments, the particles have an average diameter of about 50nm to about 500nm. In other more preferred embodiments, the particles have an average diameter of about 100nm to about 500nm. In other embodiments, the particles have an average diameter of about 500nm to about 1000nm. In certain embodiments, the particles have an average diameter of about 10nm to about 200nm, and in preferred embodiments, the particles have an average diameter of about 10nm to about 100nm. In certain embodiments, the particles have an average diameter less than 50nm. In certain embodiments, the particles have an average diameter less than 100nm. In certain embodiments, the particles have an average diameter less than 150nm. In certain embodiments, the particles have an average diameter less than 200nm. In certain embodiments, the particles have an average diameter less than 250nm. In certain embodiments, the particles have an average diameter less than 500nm.

[0166] In certain embodiments, the pharmaceutical composition further comprises a mucoadhesive. In certain embodiments, the mucoadhesive is selected from the group consisting of cellulose derivatives, polyacrylates, starch, chitosan, glycosaminoglycans, hyaluronic acid, cellulose derivatives, polyacrylates, and any combination thereof.

[0167] In certain preferred embodiments, the pharmaceutical composition further comprises a pH adjuster, an emulsifier, a pH buffer, a tonicity adjuster, a stabilizer, a preservative, a surfactant, a bulking agent, a flavoring agent, or any combination thereof.

[0168] In certain embodiments, the bulking agent is selected from mannitol, trehalose, chitosan, HP-β-cyclodextrin, hydroxypropyl methylcellulose (HPMC), dextran, starch (such as pea starch), and sucrose.

[0169] 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.

[0170] In certain embodiments, a method of preparing a pharmaceutical composition disclosed herein is provided, the method comprising:

[0171] a. dissolving one or more MPLAs in an organic solvent to form an organic solvent / MPLA solution;

[0172] b. Combining the organic solvent / MPLA solution with water to form a colloidal formulation including one or more MPLAs.

[0173] In certain embodiments, step (a) or step (b) is performed under sonication.

[0174] In certain embodiments, the organic solvent and water are present in a volume: volume ratio of about 1: 1500 to about 1: 50. In certain embodiments, the organic solvent and water are present in a volume: volume ratio of 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.

[0175] In certain embodiments, one or more surfactants are added in step (a) or step (b). In some preferred embodiments, the surfactant is carboxymethyl cellulose. In another embodiment, one or more phospholipids are added in step (a) or step (b). In certain embodiments, the mixture in step (b) is freeze-dried. 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 performed at an elevated temperature. In certain embodiments, the elevated temperature is 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.

[0176] In certain embodiments, the mixture in step (b) has a MPLA concentration ranging from about 1 μg / mL to about 1000 μg / mL; about 20 μg / mL to about 500 μg / mL, about 100 μg / mL to about 300 μg / mL, about 250 μg / mL, or about 125 μg / mL.

[0177] In certain embodiments, the spray dried powder has an MPLA concentration of 0.25% w / w MPLA / solid to 10% w / w MPLA / solid in the final powder. In certain preferred embodiments, the MPLA concentration in the final powder is 0.25% w / w to 2% w / w MPLA / solid.

[0178] In certain embodiments, the spray-dried powder is reconstituted with water prior to administration to a patient.

[0179] In some embodiments, the MPLA compound is administered in a composition that includes one or more pharmaceutically acceptable excipients. The phrase "pharmaceutically acceptable" is art-recognized. In certain embodiments, the term encompasses 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 excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0180] For example, compositions including MPLA compounds can be formulated for nasal delivery as a dry powder, as an aqueous solution, an aqueous suspension, a colloid, a water-in-oil emulsion, a micellar formulation, or as a liposomal formulation.

[0181] In some embodiments, the MPLA composition includes micelles of the MPLA compound. While not being bound by theory, it is believed that the micelles increase the activity of MPLA. In some embodiments, the size of the micelles is 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 as measured by DLS.

[0182] In certain preferred embodiments, the composition further comprises an organic solvent, such as alcohol, glycerol, 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 tert-butyl alcohol, preferably ethanol.

[0183] In some embodiments, the composition further comprises a fatty acid salt, a fatty acid, a phospholipid, or any combination thereof.

[0184] In some embodiments, the composition includes 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, lysolecithin, lysosphatidylethanolamine, cerebrosides, arachidylphosphatidylcholine (DAPC), didecanoyl-L-α-phosphatidylcholine (DDPC), di-elaidoylphosphatidylcholine (DEPC), dilauroylphosphatidylcholine (DLPC), dilinoleoylphosphatidylcholine, dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DPPC), di-stearoylphosphatidylcholine (DPPC), di-oleo ... Phosphatidylcholine (DSPC), 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), arachidonylphosphatidylglycerol (DAPG), didecanoyl-L-α-phosphatidylglycerol (DDPG), di-elaidoylphosphatidylglycerol (DEPG), dilauroylphosphatidylglycerol (DLPG), dilinoleoylphosphatidylglycerol, dimyristoylphosphatidylglycerol (DMPG), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylglycerol (DSPG), 1-palmitoyl-2-oleoylphosphatidylglycerol (POPG), arachidonylphosphatidylethanolamine (DAPE), didecanoyl-L-α-phosphatidylethanolamine (DDPE), Di-elaidoylphosphatidylethanolamine (DEPE), dilauroylphosphatidylethanolamine (DLPE), dilinoleoylphosphatidylethanolamine, dimyristoylphosphatidylethanolamine (DMPE), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylethanolamine (DSPE), 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE), arachidonylphosphatidylinositol (DAPI), didecanoyl-L-α-phosphatidylinositol (DDPI), di-elaidoylphosphatidylethanolamine (DEPI), dilauroylphosphatidylinositol (DLPI), dilinoleoylphosphatidylinositol, dimyristoylphosphatidylethanolamine (DMPI), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylethanolamine (DSPE), 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE), arachidonylphosphatidylinositol (DAPI), didecanoyl-L-α-phosphatidylinositol (DDPI), di-elaidoylphosphatidylinositol (DEPI), dilauroylphosphatidylinositol (DLPI), dilinoleoylphosphatidylinositol, dimyristoylphosphatidylinositol (DMPI), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylethanolamine (DSPE), 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE), arachidonylphosphatidylinositol (DAPI), didecanoyl-L-α-phosphati Inositol (DOPI), dipalmitoylphosphatidylinositol (DPPI), distearoylphosphatidylinositol (DSPI), 1-palmitoyl-2-oleoylphosphatidylinositol (POPI), arachidonylphosphatidylserine (DAPS), didecanoyl-L-α-phosphatidylserine (DDPS), di-antioleylphosphatidylserine (DEPS), dilauroylphosphatidylserine (DLPS), dilinoleoylphosphatidylserine, dimyristoylphosphatidylserine (DMPS), dioleoylphosphatidylserine (DOPS), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylserine (DSPS), 1-palmitoyl-2-oleoylphosphatidylserine (POPS),Arachidoyl sphingomyelin, didecanoyl 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.

[0185] In certain embodiments, the phospholipid is DPPC, DOPC, cholesterol, or a mixture thereof.

[0186] In certain embodiments, the composition comprising MPLA may contain a pH adjuster, a pH buffer, an oil / emulsifier (e.g., squalene), a tonicity adjuster, a stabilizer, a preservative, a detergent, a flavoring agent, a bulking agent, or a secondary immunostimulant. In some embodiments, the composition is a dry powder comprising a bulking agent.

[0187] Secondary immunostimulants include, for example, gonadocorticoids, deoxycholic acid, vitamin D, and beta-glucan. Suitable buffers include sodium chloride-based solutions or potassium chloride-based solutions, such as phosphate-buffered saline, potassium-buffered saline, or borate-buffered saline. In some embodiments, the buffer may contain salts, detergents, or carbohydrates that preserve MPLA when dry and help dissolve MPLA when encountering liquids. Suitable carbohydrates include trehalose, sucrose, glucose, and mannose.

[0188] In some embodiments, the composition further comprises a mucoadhesive. Suitable mucoadhesives include glycosaminoglycans (GAGS) (including chondroitin sulfate), chitosan, hyaluronic acid, cellulose derivatives, HP-B-cyclodextrin, polyacrylates, starch, HPMC, and any combination thereof.

[0189] In some embodiments, the mucoadhesive is present in the composition in an amount ranging from about 0.1% to about 50% by weight, about 25% to about 50% by weight, or about 49% by weight.

[0190] In some embodiments, the composition further comprises 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% by weight.

[0191] In some embodiments, the composition is an aqueous liquid. In such embodiments, the concentration of the MPLA compound in the composition can be about 1 μg / mL to about 1000 μg / mL, about 20 μg / mL to about 500 μg / mL, about 100 μg / mL to about 300 μg / mL, or about 250 μg / mL.

[0192] In certain embodiments, the preparation can contain ionic or nonionic surfactants. Suitable surfactants include poloxamer 407, poloxamer 181, dodecyl trimethyl ammonium bromide (DTAB), n-dodecyl octaethylene oxide (C12E8), n-dodecyl tetraethylene oxide (C12E4) and dioctanoyl phosphatidylcholine (C8-lecithin), polyethylene glycol (35) castor oil, polyoxyethylene castor oil EL (CrEL), octaethylene glycol monolauryl ether (C12E8), hexadecyl trimethyl ammonium bromide (CTAB), polypropylene oxide (PPO), polyethylene oxide (PEO), PEO-poly (D, L-lactic acid-to-caprolactone) (PEO-PDLLA) and sodium lauryl sulfate (SDS) and any combination thereof.

[0193] In some embodiments, the MPLA formulation has a pH between 4 and 9. In certain preferred embodiments, the pH is between 5 and 8.

[0194] In certain embodiments, the formulation may be free or substantially free of phospholipids, surfactants, salts (e.g., NaCl), and / or buffers. Substantially free means that the substance in question comprises less than 0.5%, less than 0.25%, less than 0.1%, less than 0.05%, less than 0.01%, or less than 0.005% by weight of the composition.

[0195] In some embodiments of the invention, the composition includes the MPLA compound at a concentration between 1 μg / mL and 8000 μg / mL. In certain preferred embodiments, MPLA is present at a concentration between 20 μg / mL and 500 μg / mL. In certain more preferred embodiments, the concentration is between 100 μg / mL and 300 μg / mL. In certain embodiments, the concentration is 250 μg / mL, and in other embodiments, the concentration is 125 μg / mL.

[0196] In some embodiments of the invention, the solution is formulated so that the surfactant is included at a concentration between 1% w / w and 40% w / w, which can enhance the absorption of the drug upon administration by preventing degradation / metabolism, enhancing barrier permeability via transient opening of tight junctions, disruption of lipid bilayer packaging / complexes / carriers / ion pairing, and increasing residence time / slowing mucociliary clearance. In certain preferred embodiments, the surfactant concentration is between 1% w / w and 25% w / w. In certain most preferred embodiments, the surfactant concentration is 15% w / w. Surfactants of interest include, but are not limited to, dipalmitoylphosphatidylcholine, soy lecithin, phosphatidylcholine, sodium taurocholate, sodium deoxycholate, sodium glycodeoxycholate, palmitic acid, stearic acid, and oleic acid.

[0197] In some embodiments of the invention, the composition includes a mucoadhesive at a concentration between 0.1% w / w and 50% w / w, which can enhance absorption of the drug upon administration by increasing residence time and / or slowing mucociliary clearance. In certain preferred embodiments, the mucoadhesive is included at a concentration between 40% w / w and 50% w / w. In certain most preferred embodiments, the mucoadhesive is included at a concentration of 49% w / w. Mucoadhesives of interest include, but are not limited to, cellulose derivatives, HP-β-cyclodextrin polyacrylate, starch, and chitosan.

[0198] In other preferred embodiments, the composition is a powder comprising an MPLA-like compound and one or more compatibilizers. Useful compositions include 2.5 to 50 wt % of an MPLA-like compound and 50 to 97.5 wt % of one or more compatibilizers. Preferred compositions contain 5 to 20 wt % of an MPLA-like compound and 80 to 95 wt % of one or more compatibilizers. Particularly preferred compositions contain about 10 wt % of an MPLA-like compound and about 90 wt % of one or more compatibilizers.

[0199] Combination therapy

[0200] A range of anti-inflammatory treatments are available for certain inflammation-related indications, such as Humira (a monoclonal antibody therapeutic that targets and removes TNF-α from the circulation). Since TNF-α is a contributing factor to inflammation, the reduction of TNF-α reduces the symptoms associated with several inflammatory conditions, such as rheumatoid arthritis, eczema, and psoriasis. Since Humira targets a single, specific pro-inflammatory cytokine, it cannot provide broad-spectrum relief for inflammation due to other sources. Monoclonal antibody treatment may take several weeks to demonstrate a reduction in symptoms.

[0201] When administered in combination with MPLA, monoclonal antibody therapy for the treatment of inflammation may provide benefits to the treatment, as it will theoretically provide both short-term and long-term reductions in symptoms caused by inflammation. In some embodiments, the monoclonal antibodies are directed to target galectin-3. In some embodiments, the monoclonal antibodies are directed to target inflammatory cytokines, including but not limited to IL-6, IL-23, IL-33, and / or IL-1β.

[0202] In some embodiments of the invention, the composition is administered prior to monoclonal antibody therapy, concurrently with monoclonal antibody therapy, or after monoclonal antibody therapy. Example

[0203] In order to more fully understand the invention described herein, the following examples are presented.The examples described in this application are provided to illustrate the compounds, compositions, materials, devices and methods provided herein, and should not be construed in any way to limit the scope thereof.

[0204] Example 1

[0205] Preparation of PHAD micelles in 5% ethanol.

[0206] 1 mg of PHAD was soaked 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 water was added QS to 8 ml to obtain a uniform preparation of PHAD micelles with a size of 150 nm or less. The preparation can be used as a liquid or lyophilized agent.

[0207] Example 2

[0208] Preparation of PHAD powder.

[0209] At 40°C, 6 mg of PHAD was wetted and dissolved in 3 ml of 95% ethanol and sonicated at 40°C for 20 minutes until a clear solution of 2 mg / mL PHAD was obtained. 17 mL of water was then added at 40°C and sonicated to thoroughly mix the bulk solution to obtain a uniform preparation of PHAD micelles with a size of approximately 150 nm or less. 147 mg of HP-β-cyclodextrin and 147 mg of trehalose dihydrate were added to the solution under mixing conditions. The solution was then spray dried to obtain the final powder: 2% w / w PHAD: 49% w / w HP-β-cyclodextrin: 49% w / w trehalose.

[0210] The powder is stable for extended periods of time at ambient temperature and is readily soluble in purified water at concentrations up to about 5-10 mg / mL. The powder can be reconstituted for parenteral administration, such as by intravenous, subcutaneous or intramuscular routes of administration, or by intranasal administration. Alternatively, the same powder can be formulated with additional excipients to form tablets or gels for oral delivery.

[0211] Example 3

[0212] The formulations prepared in Examples 1 and 2 showed robust upregulation of IP-10 due to in vitro stimulation of TLR4 in mouse macrophages ( Figure 2). IP-10 is an important cytokine associated with the stimulation of the TRIF pathway. The evidence of IP-10 upregulation confirms that the PHAD formulations mentioned in this application are able to selectively stimulate the TRIF pathway, which further confirms the reduction or decrease of activity mediated by the MyD88 signaling pathway, which supports the reduction of pro-inflammatory cytokine production of MPLA as a treatment for inflammatory conditions.

[0213] Example 4

[0214] In validated preclinical models of acute kidney injury (AKI) and chronic kidney disease (CKD), daily administration of REVTx-300, an MPLA-like compound, resulted in significant reductions in fibrosis and circulating transforming growth factor β (TGF-β) in a dose-dependent manner relative to positive control groups ( Figure 3 and Figure 4 ). Composite data represent the average of 3 anatomically distinct depths (10 images / depth / rat / group = ~60-65% of renal cortical area). Renal cortical fibrosis, expressed as collagen volume fraction (CVF; quantification via PSR-stained tissue sections), was increased in vehicle-treated UUO-obstructed kidneys relative to sham controls. SB-525334 attenuated the UUO-induced increase in renal cortical CVF. REVTx-300 (PHAD and MPLA-like compound) administered at 0.3 mg / kg and 0.9 mg / kg attenuated the UUO-induced increase in renal cortical CVF. In addition, in the unilateral ureteral obstruction (UUO) model, REVTx-300 significantly increased circulating anti-inflammatory interleukin-10 (IL-10) in a dose-dependent manner in all groups relative to the positive control group ( Figure 8 )、Hepcidin( Figure 6 ) and neutrophil gelatinase-associated lipocalin (NGAL) ( Figure 5 ).

[0215] TGF-β is a key driver of fibrogenesis and directly promotes collagen deposition through overproduction of extracellular matrix. IL-10 is described as an anti-inflammatory cytokine because it is able to reduce the production of pro-inflammatory mediators. Hepcidin and NGAL sequester iron to prevent iron-mediated reactive oxygen species tissue damage. There was no significant increase in markers of inflammation (IL-6 was not increased, IL-1β ( Fig. 9 ) and IL-18( Fig.10 ) were relatively small increases. These results provide mechanistic evidence for the reduction in fibrosis observed in the UUO model in response to treatment with REVTx-300.

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Claims

1. A method for treating or preventing organ function loss due to chronic organ disease by administering an effective amount of a Toll-like receptor 4 (TLR4) agonist to a subject in need thereof.

2. The method of claim 1, wherein the TLR4 agonist is an MPLA-like compound.

3. The method of claim 1 or 2, wherein the TLR4 is a phosphorylated hexaacyl disaccharide (PHAD), a 3-deacylated phosphorylated hexaacyl disaccharide (3D-PHAD), a 3D-(6-acyl)phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or any combination thereof, or a pharmaceutically acceptable salt thereof. The method of claim 3 , wherein the TLR4 agonist is PHAD or a pharmaceutically acceptable salt thereof.

5. The method of any one of claims 2 to 4, wherein the MPLA-like compound is administered as an aqueous solution.

6. The method of any one of claims 2 to 5, wherein the MPLA-like compound is delivered parenterally as an aqueous solution.

7. The method of any one of claims 2 to 4, wherein the MPLA-like compound is delivered orally as a tablet.

8. The method of any one of claims 1 to 7, wherein the TLR4 agonist selectively stimulates the TIR domain clustering adaptor-inducing interferon-β (TRIF) pathway.

9. The method of any one of claims 1 to 8, wherein the chronic organ disease is selected from the group consisting of non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, osteoarthritis, rheumatoid arthritis, irritable bowel syndrome, fibrotic pulmonary disease, heart disease, and any combination thereof.

10. The method of any one of claims 1 to 9, wherein the TLR4 agonist reduces circulating TGF-β in the subject.

11. The method of any one of claims 1 to 10, wherein the TLR4 agonist increases circulating interleukin-10 (IL-10), circulating hepcidin, and / or circulating neutrophil gelatinase-associated lipocalin (NGAL) in the subject.

12. The method of any one of claims 1 to 11, wherein the method prevents loss of organ function due to chronic organ disease.

13. The method of any one of claims 1 to 12, wherein the method treats loss of organ function due to chronic organ disease.

14. 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 thereof.

15. The method of claim 14, wherein the TLR4 agonist is an MPLA compound.

16. The method of claim 14 or 15, wherein the TLR4 agonist is a phosphorylated hexaacyl disaccharide (PHAD), a 3-deacylated phosphorylated hexaacyl disaccharide (3D-PHAD), a 3D-(6-acyl)phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or any combination thereof, or a pharmaceutically acceptable salt thereof.

17. The method of claim 16, wherein the TLR4 agonist is PHAD or a pharmaceutically acceptable salt thereof.

18. The method of any one of claims 15 to 17, wherein the MPLA-like compound is administered as an aqueous solution.

19. The method of any one of claims 15 to 18, wherein the MPLA-like compound is delivered parenterally as an aqueous solution.

20. The method of any one of claims 15 to 17, wherein the MPLA-like compound is delivered orally as a tablet.

21. The method of any one of claims 14 to 20, wherein the loss of renal function is associated with one or more of: a Type I or II diabetes, hypertension, glomerulonephritis, polycystic kidney disease, autoimmune disease, vesicoureteral reflux, pyelonephritis, interstitial nephritis, kidney stones, kidney obstruction, cancer, substance abuse, kidney inflammation, kidney fibrosis, medication overuse, and chemotherapy.

22. The method of any one of claims 14 to 21, wherein the TLR4 agonist selectively stimulates the TIR domain clustering adaptor-inducing interferon-β (TRIF) pathway.

23. The method of any one of claims 14 to 22, wherein the TLR4 agonist reduces circulating TGF-β in the subject.

24. The method of any one of claims 14 to 22, wherein the TLR4 agonist increases circulating interleukin-10 (IL-10), circulating hepcidin, and / or circulating neutrophil gelatinase-associated lipocalin (NGAL) in the subject.

25. The method of any one of claims 14 to 24, wherein the method prevents loss of kidney function due to chronic kidney disease.

26. The method of any one of claims 14 to 25, wherein the method treats loss of kidney function due to chronic kidney disease.

27. 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 thereof.

28. The method of claim 27, wherein the TLR4 agonist is an MPLA compound.

29. The method of claim 27 or 28, wherein the TLR4 agonist is a phosphorylated hexaacyl disaccharide (PHAD), a 3-deacylated phosphorylated hexaacyl disaccharide (3D-PHAD), a 3D-(6-acyl)phosphorylated hexaacyl disaccharide (3D(6-acyl)PHAD) or any combination thereof, or a pharmaceutically acceptable salt thereof.

30. The method of claim 29, wherein the TLR4 agonist is PHAD or a pharmaceutically acceptable salt thereof.

31. The method of any one of claims 28 to 30, wherein the MPLA-like compound is administered as an aqueous solution.

32. The method of any one of claims 28 to 31, wherein the MPLA-like compound is delivered parenterally as an aqueous solution.

33. The method of any one of claims 28 to 30, wherein the MPLA-like compound is delivered orally as a tablet.

34. The method of any one of claims 27 to 33, wherein the organ is a kidney.

35. The method of any one of claims 27 to 34, wherein the acute stress is due to one or more of the following: a. Toxicity from drugs, toxicity from chemotherapy, ischemia, trauma, cancer, or infection.

36. The method of any one of claims 27 to 35, wherein the TLR4 agonist selectively stimulates the TIR domain clustering adaptor-inducing interferon-β (TRIF) pathway.

37. The method of any one of claims 27 to 36, wherein the TLR4 agonist reduces circulating TGF-β in the subject.

38. The method of any one of claims 27 to 37, wherein the TLR4 agonist increases circulating interleukin-10 (IL-10), circulating hepcidin, and / or circulating neutrophil gelatinase-associated lipocalin (NGAL) in the subject.

39. The method of any one of claims 27 to 38, wherein the loss of organ function is due to chronic inflammation or fibrosis.

40. The method of any one of claims 27 to 39, wherein the method prevents loss of organ function due to acute stress.

41. The method of any one of claims 27 to 40, wherein the method treats loss of organ function due to acute stress.

42. A pharmaceutical composition for treating or preventing loss of organ function in a subject in need thereof, the pharmaceutical composition comprising a colloidal formulation of a monophosphoryl lipid A (MPLA)-like compound.

43. The pharmaceutical composition of claim 42, wherein the MPLA-like compound is selected from phosphorylated hexaacyl disaccharide (PHAD), PHAD-504, 3D-(6-acyl)-PHAD, 3D-PHAD, and any combination thereof.

44. The pharmaceutical composition of claim 43, wherein the MPLA-like compound is PHAD.

45. A pharmaceutical composition as described in any one of claims 43 to 45, wherein the pharmaceutical composition is an aqueous composition.

46. ​​The pharmaceutical composition of any one of claims 42 to 45, wherein the pharmaceutical composition is a dry powder.

47. The pharmaceutical composition of claim 45, wherein the pharmaceutical composition has a MPLA concentration of about 1 μg / mL to about 10000 μg / mL.

48. A pharmaceutical composition as described in any one of claims 42 to 47, further comprising a stabilizer.

49. The pharmaceutical composition of claim 48, wherein the stabilizer is trehalose.

50. The pharmaceutical composition of any one of claims 42 to 49, wherein the composition comprises micelles having an average diameter or length of about 1 nm to about 1000 nm.

51. The pharmaceutical composition of any one of claims 42 to 50, further comprising an 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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