Compositions and methods for promoting brain health

Abstract

Compositions and methods for promoting brain health are provided. The present disclosure provides a composition comprising an active ingredient encapsulated in a suspension of particles comprising a biodegradable polymer containing an effective amount of the active ingredient, the particles being configured to bypass the blood-brain barrier, the nutritional supplement being synthetic D-beta hydroxybutyrate, and the targeted delivery being to the brain. Also provided are methods for promoting brain healing after traumatic brain injury, including neuroinflammation, and methods for promoting brain health. Further provided is a pharmaceutical composition for use in reducing the risk of brain damage due to acquired brain injury in a subject susceptible to or otherwise at risk for acquired brain injury, the composition comprising a pharma- ceutically acceptable carrier and a formulation comprising a therapeutic amount of an API, where the API is synthetic D-beta hydroxybutyrate, and the treatment comprises targeted delivery to the olfactory region of the subject's nasal cavity by intranasal administration of the composition (a) before the subject participates in an event where acquired brain injury is a known risk, (b) after the subject participates in the event, or both, for use in treating a symptom of acquired brain injury in a subject, including neuroinflammation, and wherein the treatment is neuroprotective.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 316,845, entitled “COMPOSITIONS AND METHODS TO PROMOTE BRAIN HEATH,” filed March 4, 2022, the entire contents of which are incorporated herein by reference in their entirety. [Background technology]

[0002] Biology of Wound Healing Wounds result from injury or disruption to normal anatomical structures and functions (Robson MC et al., Curr Probl Surg 2001;38:72-140; Velnar T et al., The Journal of International Medical Research 2009;37:1528-1542). This can range from simple disruption of the epithelial integrity of the skin to deeper subcutaneous tissues with damage to other structures including tendons, muscles, blood vessels, nerves, solid organs, and even bone (Alonso JE et al., Surg Clin North Am 1996;76:879-903). Regardless of cause and form, wounds damage and disrupt the local tissue environment.

[0003] Wound healing is a dynamic, interactive process involving soluble mediators, blood cells, extracellular matrix, and parenchymal cells. The wound repair process can be divided into four phases that overlap in time and space: (1) the coagulation phase, (2) the inflammatory phase, (3) the proliferation phase, and (4) the remodeling phase. The immune system is usually an active participant in wound healing. Much of what is known is based on human skin wound healing.

[0004] Solidification stage Immediately after injury, platelets attach to damaged blood vessels and initiate a release response, initiating the hemostatic response, preventing excessive bleeding, and triggering the blood coagulation cascade that provides provisional protection to the injured area. Platelets release well over a dozen growth factors, cytokines, and other survival or apoptosis inducers (Weyrich AS and Zimmerman GA, Trends Immunol 2004 Sep;25(9):489-495). Major components of the platelet release response include platelet-derived growth factor (PDGF) and transforming growth factors A1 and 2 (TGF-A1 and TGF-2), which attract inflammatory cells such as leukocytes, neutrophils, and macrophages (Singer AF and Clark RA, N Engl J Med 1999 Sep 2;341(10):738-746).

[0005] Inflammation stage The inflammatory phase is triggered by capillary injury, leading to the formation of a clot / provisional matrix composed of fibrin and fibronectin. This provisional matrix fills the tissue defect and allows the influx of effector cells. Platelets present in the clot release multiple cytokines involved in the recruitment of inflammatory cells (neutrophils, monocytes, and macrophages, among others), fibroblasts, and endothelial cells (EC).

[0006] Multiplication stage The inflammatory phase is followed by a proliferative phase, in which active angiogenesis generates new capillaries, allowing the delivery of nutrients to the wound site and supporting, among other things, the proliferation of fibroblasts. Fibroblasts present in the granulation tissue become activated and acquire a smooth muscle cell-like phenotype, and are then called myofibroblasts. Myofibroblasts synthesize and deposit components of the extracellular matrix (ECM) that replace the provisional matrix. Myofibroblasts also have contractile properties mediated by α-smooth muscle actin organized into microfilament bundles or stress fibers. Myofibroblast differentiation of fibroblasts begins with the appearance of protomyofibroblasts, whose stress fibers contain only β- and γ-cytoplasmic actin. Protomyofibroblasts can evolve into differentiated myofibroblasts, whose stress fibers contain α-smooth muscle actin.

[0007] Remodeling Phase The fourth healing stage involves the gradual remodeling and re-epithelialization of granulation tissue. This remodeling process is mainly mediated by proteolytic enzymes, especially matrix metalloproteinases (MMPs) and their inhibitors (TIMPs, tissue inhibitors of metalloproteinases). During re-epithelialization, type III collagen, the main component of granulation tissue, is gradually replaced by type I collagen, the main structural component of the dermis. Elastin, which contributes to skin elasticity and is absent in granulation tissue, also reappears. Cell density is normalized (resolution) by apoptosis of vascular cells and myofibroblasts.

[0008] inflammation Tissue injury causes the destruction of blood vessels and the extravasation of blood components. Blood clots re-establish hemostasis and provide a provisional extracellular matrix for cell migration. Platelets not only promote the formation of a hemostatic plug but also secrete several mediators of wound healing (e.g., platelet-derived growth factors) that attract and activate macrophages and fibroblasts (Heldin, C. and Westermark B., In: Clark R., ed. The molecular and cellular biology of wound repair, 2nd Ed. New York, Plenum Press, pp. 249-273, (1996)). However, in the absence of bleeding, it has been suggested that platelets are not essential for wound healing. Numerous vasoactive mediators and chemotactic factors are produced by the coagulation and activated complement pathways and by injured or activated parenchymal cells that have been shown to recruit inflammatory leukocytes to the site of injury (ibid.).

[0009] Cellular entry into the wound and activation of local cells is initiated by mediators either released de novo by resident cells or from stored residues in the granules of platelets and basophils. Sephel, GC and Woodward, SC, 3. Repair, Regeneration and Fibrosis,” in Rubin's Pathology, Rubin, R. and Strayer, DSEds; 5th Ed., Wolters Kluwyer Health, / Lippincott Williams & Wilkins, Philadelphia, PA (2008), at 71. Cell migration employs the cellular response to cytokines and insoluble substrates of the extracellular matrix. Ibid. at 72.

[0010] Infiltrating neutrophils cleanse the wounded area of ​​foreign particles and bacteria, then are extruded along with eschar (dead tissue that is sloughed off (sloughed off) from healthy skin or phagocytosed by macrophages). In response to specific chemoattractants, such as fragments of extracellular matrix proteins, transforming growth factor beta (TGF-β), and monocyte chemoattractant protein-1 (MCP-1), monocytes also infiltrate the wound site and become activated macrophages that release growth factors (e.g., platelet-derived growth factor and vascular endothelial growth factor) that initiate the formation of granulation tissue. Macrophages bind to specific proteins of the extracellular matrix by their integrin receptors, an action that stimulates the phagocytosis of microorganisms and fragments of the extracellular matrix by macrophages (Brown, E. Phagocytosis, Bioessays, 17:109-117 (1995)). Studies have also reported that adhesion to extracellular matrix stimulates monocytes to undergo transformation into inflammatory or repair macrophages. These macrophages play a key role in the transition between inflammation and repair (Riches, D., In Clark R., Ed. The molecular and cellular biology of wound repair, 2nd Ed. New York, Plenum Press, pp.95-141). For example, adhesion causes monocytes and macrophages to express colony-stimulating factor-1 (CSF-1), a cytokine required for monocyte and macrophage survival, tumor necrosis factor-α (TNF-α), a potent inflammatory cytokine, and platelet-derived growth factor (PDGF), a potent chemoattractant and mitogen for fibroblasts. Other cytokines shown to be expressed by monocytes and macrophages include transforming growth factor (TGF-α), interleukin-1 (IL-1), transforming growth factor β (TGF-β), and insulin-like growth factor-I (IGF-I) (Rappolee, D. et al., Science, 241, pp. 708-712 (1988)).It has been suggested that monocyte- and macrophage-derived growth factors are required for the initiation and proliferation of new tissue formation in wounds, as macrophage-depleted animals have defective wound repair (Leibovich, S, and Ross, R., Am J Pathol, 78, pp1-100 (1975)).

[0011] epithelialization Wound re-epithelialization begins within hours after injury. Epithelial cells from skin appendages such as hair follicles rapidly remove clotted blood and damaged stroma from the wound space. At the same time, cells undergo phenotypic changes including the retraction of intracellular tonofilaments (Paladini, R. et al., J. Cell Biol, 132, pp. 381-397 (1996)), dissolution of most intercellular desmosomes that provide physical connections between cells, and the formation of peripheral cytoplasmic actin filaments that allow cell movement and migration (Goliger, J. and Paul, D. Mol Biol Cell, 6, pp. 1491-1501 (1995); Gabbiani, G. et al., J Cell Biol, 76, PP. 561-568 (1978)). Furthermore, epidermal and dermal cells no longer adhere to each other due to the dissolution of hemidesmosomal junctions between the epidermis and the basement membrane, allowing lateral migration of epidermal cells. The expression of integrin receptors on epidermal cells allows them to interact with various extracellular matrix proteins (e.g., fibronectin and vitronectin) that are interspersed with interstitial type I collagen at the wound edge and interwoven with the fibrin clot in the wound space (Clark, R., J Invest Dermatol, 94, Suppl, pp. 128S-134S (1990)). Migrating epidermal cells cut the wound and separate the dry eschar (dead tissue that is sloughed (sloughed) from healthy skin) from viable tissue. The path of cutting appears to be determined by a series of integrins that the migrating epidermal cells express on their cell membrane.

[0012] The degradation of the extracellular matrix required for epidermal cell migration between the collagenous dermis and the fibrin eschar depends on the production of collagenase by epidermal cells (Pilcher, B. et al., J Cell Biol, 137, pp. 1445-1457 (1997)) and on the activation of plasmin by plasminogen activator produced by epidermal cells (Bugge, T. et al., Cell, 87, 709-719 (1996)). Plasminogen activator also activates collagenase (matrix metalloproteinase-1) (Mignatti, P. et al., Proteinases and Tissue Remodeling. In Clark, R. Ed. The molecular and cellular biology of wound repair. 2nd Ed. New York, Plenum Press, 427-474 (1996)), promoting the degradation of collagen and extracellular matrix proteins.

[0013] One to two days after injury, epithelial cells at the wound edge begin to proliferate behind the actively migrating cells. The stimuli for epidermal cell migration and proliferation during re-epithelialization remain to be determined, but several possibilities have been suggested. The absence of adjacent cells at the wound edge (the "free edge" effect) may signal both epidermal cell migration and proliferation. Local release of growth factors and increased expression of growth factor receptors may also stimulate these processes. The leading candidates include epidermal growth factor (EGF), transforming growth factor-α (TGF-α), and keratinocyte growth factor (KGF) (Nanney, L. and King, L. Epidermal Growth Factor and Transforming Growth Factor-α. In Clark, R. Ed. The molecular and cellular biology of wound repair. 2nd Ed. New York, Plenum Press, pp. 171-194 (1996); Werner, S. et al., Science, 266, pp. 819-822 (1994); Abraham, J. and Klagsburn, M. Modulation of Wound Repair by Members of the Fibroblast Growth Factor family. In Clark, R. Ed. The molecular and cellular biology of wound repair. 2nd Ed. New York, Plenum Press, pp. 195-248 (1996)). As re-epithelialization continues, basement membrane proteins reappear in a highly ordered, zipper-like arrangement from the wound edge inwards (Clark R. et al., J. Invest Dermatol, 79, pp. 264-269 (1982)). Epidermal cells revert to their normal phenotype and again firmly attach to the re-established basement membrane and underlying dermis.

[0014] Granulation tissue formation New stroma, often called granulation tissue, begins to invade the wound space about four days after injury. Numerous new capillaries give the new stroma its granular appearance. Macrophages, fibroblasts, and blood vessels migrate simultaneously into the wound space (Hunt, T. ed. Wound Healing and Wound Infection: Theory and Surgical Practice. New York, Appleton-Century-Crofts (1980)). Macrophages provide a continuous source of growth factors necessary to stimulate fibrosis and angiogenesis, fibroblasts produce new extracellular matrix necessary to support cell growth, and blood vessels deliver oxygen and nutrients necessary to maintain cell metabolism.

[0015] Growth factors, particularly platelet-derived growth factor-4 (PDGF-4) and transforming growth factor beta-1 (TGF-β1) (Roberts, A. and Sporn, M, Transforming Growth Factor-1, In Clark, R. ed. The molecular and cellular biology of wound repair. 2nd Ed. New York, Plenum Press, pp. 275-308 (1996)), in conjunction with extracellular matrix molecules (Gray, A. et al., J Cell Sci, 104, pp. 409-413 (1993); Xu, J. and Clark, R., J Cell Biol, 132, pp. 239-149 (1996)), have been shown to stimulate fibroblasts in the tissue surrounding the wound to proliferate, express appropriate integrin receptors, and migrate into the wound space. It has been reported that platelet-derived growth factor accelerates the healing of chronic pressure sores (Robson, M. et al., Lancet, 339, pp. 23-25 ​​(1992) and diabetic ulcers (Steed, D., J Vasc Surg, 21, pp. 71-78 (1995)). In some other cases, basic fibroblast growth factor (bFGF) has been effective in treating chronic pressure sores (Robson, M. et al., Ann Surg, 216, pp. 401-406 (1992)).

[0016] The structural molecules of the newly formed extracellular matrix are called the provisional matrix (Clark, R. et al., J. Invest Dermatol, 79, pp. 264-269, 1982) and contribute to the formation of granulation tissue by providing a scaffold or conduit for cell migration. These molecules include fibrin, fibronectin, and hyaluronic acid (Greiling, D. and Clark R., J. Cell Sci, 110, pp. 861-870 (1997)). The appearance of fibronectin and appropriate integrin receptors that bind to fibronectin, fibrin, or both on fibroblasts has been suggested to be a rate-limiting step in the formation of granulation tissue. Fibroblasts are involved in the synthesis, deposition, and remodeling of the extracellular matrix, which itself can have positive or negative effects on the ability of fibroblasts to perform these tasks and generally interact with their environment (Xu, J. and Clark, R., J Cell Sci, 132, pp. 239-249 (1996); Clark, R. et al., J Cell Sci, 108, pp. 1251-1261).

[0017] Migration of cells into cross-linked fibrin clots or into tightly woven extracellular matrices requires an active proteolytic system capable of cleaving pathways for cell migration. In addition to serum-derived plasmin, various fibroblast-derived enzymes have been suggested to be potential candidates for this task, including plasminogen activators, collagenases, gelatinase A, and stromelysin (Mignatti, P. et al., Proteinases and Tissue Remodeling. In Clark, R. Ed. The molecular and cellular biology of wound repair. 2nd Ed. New York, Plenum Press, 427-474 (1996); Vaalamo, M. et al., J Invest Dermatol, 109, pp. 96-101 (1997)). After migration into the wound, fibroblasts begin to synthesize the extracellular matrix. The provisional extracellular matrix is ​​gradually replaced by a collagenous matrix, presumably in response to transforming growth factor-β1 (TGF-β1) signaling (Clark, R. et al., J Cell Sci, 108, pp. 1251-1261 (1995); Welch, M. et al., J. Cell Biol, 110, pp. 133-145 (1990)).

[0018] Once a rich collagen matrix is ​​deposited in the wound, fibroblasts cease production of collagen, and the fibroblast-rich granular tissue is replaced by a relatively acellular scar. Cells within the wound undergo apoptosis triggered by unknown signals. Dysregulation of these processes has been reported to occur in fibrotic disorders such as keloid formation, hypertrophic scars, dysmorphic scars, and scleroderma.

[0019] angiogenesis The formation of new blood vessels (angiogenesis) is necessary to maintain the newly formed granulation tissue. Angiogenesis is a complex process that depends on the extracellular matrix in the wound bed and on migration and mitogenic stimuli of endothelial cells (Madri, J. et al., Angiogenesis in Clark, R. Ed. The molecular and cellular biology of wound repair. 2nd Ed. New York, Plenum Press, pp. 355-371 (1996)). Induction of angiogenesis was initially attributed to acidic or basic fibroblast growth factors. Later, many other molecules, including vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β), angiogenin, angiotropin, angiopoietin-1, and thrombospondin, have also been found to have angiogenic activity (Folkman, J. and D'Amore, P, Cell, 87, pp. 1153-1155 (1996)).

[0020] These molecules induce angiogenesis by stimulating the production of basic fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) by macrophages and endothelial cells. For example, it has been reported that activated epithelial cells in wounds secrete large amounts of vascular endothelial growth factor (VEGF) (Brown, L. et al., J Exp Med, 176, 1375-1379 (1992)).

[0021] Basic fibroblast growth factor has been hypothesized to set the stage for angiogenesis during the first 3 days of wound repair, while vascular endothelial growth factor is important for angiogenesis during the formation of granulation tissue on days 4-7 (Nissen, N. et al., Am J Pathol, 152, 1445-1552 (1998)).

[0022] In addition to angiogenic factors, it has been shown that an appropriate extracellular matrix and endothelial receptors for a provisional matrix are necessary for angiogenesis. Microvascular endothelial cells adjacent to and growing within a wound temporarily deposit increased amounts of fibronectin in the vessel wall (Clark, R. et al., J. Exp Med, 156, 646-651 (1982)). Because angiogenesis requires the expression of functional fibronectin receptors by endothelial cells (Brooks, P. et al., Science, 264, 569-571 (1994)), it has been suggested that perivascular fibronectin serves as a conduit for the migration of endothelial cells into the wound. In addition, protease expression and activity have also been shown to be necessary for angiogenesis (Pintucci, G. et al., Semin Thromb Hemost, 22, 517-524 (1996)).

[0023] The sequence of events leading to angiogenesis has been proposed as follows: Injury causes tissue destruction and hypoxia. Angiogenic factors such as acidic and basic fibroblast growth factors (FGFs) are released from macrophages soon after cell destruction, and production of vascular endothelial growth factor by epidermal cells is stimulated by hypoxia. Proteolytic enzymes released into connective tissues degrade extracellular matrix proteins. Fragments of these proteins recruit peripheral blood monocytes to the site of injury, where they become activated macrophages and can release angiogenic factors. Certain macrophage angiogenic factors, such as basic fibroblast growth factor (bFGF), stimulate endothelial cells to release plasminogen activators and procollagenase. Plasminogen activators convert plasminogen to plasmin and procollagenase to active collagenase, and these two proteases work together to digest the basement membrane. Fragmentation of the basement membrane allows endothelial cells stimulated by angiogenic factors to migrate and form new blood vessels at the site of injury. Once the wound is filled with new granulation tissue, angiogenesis ceases and many of the new blood vessels collapse as a result of apoptosis (Ilan, N. et al., J Cell Sci, 111, 3621-3631 (1998)). It has been suggested that this programmed cell death is regulated by various matrix molecules such as thrombospondin 1 and 2, and antiangiogenic factors such as angiostatin, endostatin, and angiopoietin 2 (Folkman, J., Angiogenesis and angiogenesis inhibition: an overview, EXS, 79, 1-8, (1997)).

[0024] Wound contraction and extracellular matrix reorganization Wound contraction involves a complex and coordinated interplay of cells, extracellular matrix, and cytokines. During the second week of healing, fibroblasts assume a myofibroblast phenotype characterized by large bundles of actin-containing microfilaments arranged along the cytoplasmic face of the plasma membrane of the cells and by cell-cell and cell-matrix junctions (Welch, M. et al., J Cell Biol, 110, 133-145 (1990); Desmouliere, A. and Gabbiani, G. The role of the myofibroblast in wound healing and fibrocontractive diseases. In Clark, R. Ed. The molecular and cellular biology of wound repair. 2nd Ed. New York, Plenum Press, pp. 391-423 (1996)). The appearance of myofibroblasts corresponds to the onset of connective tissue compression and wound contraction. This contraction has been suggested to require stimulation with transforming growth factor (TGF)-β1 or β2 and platelet-derived growth factor (PDGF), binding of fibroblasts to the collagen matrix via integrin receptors, and cross-linking between individual collagen bundles. (Montesano, R. and Orci, Proc Natl Acad Sci USA, 85, 4894-4897 (1988); Clark, R. et al., J Clin Invest, 84, 1036-1040 (1989); Schiro, J. et al., Cell, 67, 403-410 (1991); Woodley, D. et al., J Invest Dermatol, 97, 580-585 (1991)).

[0025] Collagen remodeling during the transition from granulation tissue to scar depends on the continued synthesis and catabolism of collagen at a low rate. Collagen degradation in wounds is controlled by several proteolytic enzymes called matrix metalloproteinases (MMPs) secreted by macrophages, epithelial and endothelial cells, and fibroblasts (Mignatti, P. et al., Proteinases and Tissue Remodeling. In Clark, R. Ed. The molecular and cellular biology of wound repair. 2nd Ed. New York, Plenum Press, 427-474 (1996)). It has been suggested that the various stages of wound repair depend on distinct combinations of matrix metalloproteinases and tissue inhibitors of metalloproteinases (Madlener, M. et al., Exp Cell Res, 242, 201-210 (1998)).

[0026] Wounds only gain about 20% of their ultimate strength in the first 3 weeks, during which time fibrillar collagen accumulates relatively rapidly and remodels by wound contraction. Thereafter, wounds gain tensile strength at a slower rate, reflecting a much slower rate of collagen accumulation and collagen remodeling by the formation of larger collagen bundles and an increased number of intermolecular crosslinks.

[0027] Acquired brain injury Acquired brain injury is brain injury caused by events after birth, not as part of genetic or congenital disorders. Examples of acquired brain injury include, but are not limited to, traumatic brain injury (TBI), stroke, brain tumor, poisoning, infection and disease, drowning or other anoxic episodes, and alcohol and drug abuse. Examples of situations that cause acquired brain injury may include, but are not limited to, domestic violence, falls, car accidents, etc. Acquired brain injury is different from degenerative brain conditions such as Alzheimer's disease or Parkinson's disease.

[0028] Neurodegenerative diseases Neurodegenerative diseases occur when nerve cells in the brain or peripheral nervous system lose function over time and eventually die. Although certain treatments can help alleviate some of the physical or mental symptoms associated with neurodegenerative diseases, their progression cannot currently be slowed and there is no cure. Examples include, but are not limited to, Parkinson's disease, Alzheimer's disease and other memory disorders, Huntington's disease, motor neuron disease or amyotrophic lateral sclerosis (ALS), ataxia, multiple atrophy, and progressive supranuclear palsy. As they progress, these disorders can affect all aspects of people's lives, including mobility and balance, abnormal movements; swallowing; bladder and bowel function, blood pressure fluctuations; sleep; breathing; heart function; memory and cognitive abilities; mood and speech. It has been hypothesized that exposure to environmental contaminants, such as pesticides, fungicides, insecticides, metals (e.g., arsenic, lead, manganese), chemicals (e.g., polychlorinated biphenyls), air pollution, biological agents (e.g., bacterial endotoxins), dietary and lifestyle factors (e.g., caffeine, tobacco smoke, dietary antioxidants), either individually or in combination with specific genes, may influence neurodegeneration.

[0029] For example, Parkinson's disease progresses slowly as small clusters of brain neurons die. The gradual loss of these neurons reduces levels of the neurotransmitter dopamine, which transmits messages to the part of the brain that coordinates muscle movement. Common symptoms of Parkinson's disease include tremors or shaking of the hands, arms, legs, jaw, and face; stiffness or stiffness of the limbs and trunk; bradycardia, or slowness of movement; and difficulties with balance, speech, and coordination. Symptoms begin gradually and typically worsen over time. Scientists do not know the cause of Parkinson's disease, but most agree that interactions between a person's genes and environment play a role in the development and progression of the disease.

[0030] Alzheimer's disease (AD) is a progressive neurodegenerative disorder, characterized in most cases by early memory impairment and progressive and irreversible cognitive decline that may ultimately affect behavior, speech, visuospatial orientation, and the motor system. Pathologically, AD is defined by two lesions evident on microscopic examination of postmortem tissue: extracellular plaques composed primarily of 1-42 amino acid polypeptides of aggregated amyloid-β (Aβ), and intracellular neurofibrillary tangles (NFTs) formed from hyperphosphorylated aggregated forms of microtubule-associated tau [Montine, TJ et al. Acta Neuropathol. (2012) 123(1):1-11]. These proteins aggregate into highly toxic soluble oligomeric structures and ultimately into microscopically detectable fibrillar macrostructures such as plaques and tangles. This is associated with synaptic dysfunction and loss in areas where one or both protein aggregates are found, disruption of neurotransmitter systems, and neuronal cell death (Selkoe, DJ, Science (2002) 298(5594):789-91; Spires-Jones, TL & Hyman, BT. Neuron (2014) 82(4):756-71). Ultimately, neurodegeneration leads to cognitive decline and dementia [Hardy, J. and Selkoe, DJ. Science (2002) 297(5580):353-6].

[0031] Huntington's disease (HD) is a rare progressive neurodegenerative disorder with autosomal dominant inheritance. HD is caused by mutations in the gene for the protein huntingtin, which causes a CAG trinucleotide (cytosine, adenine, and guanine) repeat expansion located in the first exon of the HD gene, which encodes huntingtin, a 350 kDa protein of unknown function. Expansion of CAG trinucleotide repeats (CAG repeats) in the coding region of the human gene causes neurodegenerative disorders by generating proteins with elongated polyglutamine (polyQ) expansions. This group of disorders includes Huntington's disease (HD), dentatorubral-pallidoluysian atrophy, spinal-bulbar muscular atrophy, and spinocerebellar degeneration (SCA) types 1, 2, 3, 6, and 7 [Moncke-Buchner, E. et al. Nucleic Acid Res. (2002) 30(16):e83].

[0032] The term "motor neuron disease" describes a heterogeneous group of relentlessly progressive and ultimately fatal conditions (including amyotrophic lateral sclerosis, or ALS) that cause progressive muscle weakness through loss of upper and lower motor neurons. Non-motor pathways are also affected, with up to 50% of patients having detectable cognitive and behavioral changes. [Verber, NS, et al. Front. Neurol. (2019) 10:291, citing Wooley, SC and Jonathan, SK. Phys. Med. Rehabil. Clin. N. Am. (2008) 19:607-17].

[0033] Ataxia, or lack of coordination, is a common manifestation of a variety of neurological conditions, including stroke, brain tumors, multiple sclerosis, traumatic brain injury, toxicity, infection (including after chickenpox), and congenital cerebellar defects. Its evolution may be acute, subacute, recurrent paroxysmal, or chronic. Ataxias may be broadly divided into those that are genetic (with or without family history) and those that are acquired / degenerative. "Spora" ataxia means there is no family history. Acquired progressive ataxias may be immune-mediated (e.g., paraneoplastic spinocerebellar degeneration, gluten ataxia), degenerative (e.g., cerebellar variant of multiple system atrophy (type C), caused by deficiency states (e.g., vitamin B12, vitamin E, etc.), toxic (e.g., alcohol-associated ataxia, phenytoin), or infection-related (HIV, sporadic Creutzfeldt-Jakob disease, progressive multifocal leukoencephalopathy, etc.). Genetic ataxias may have autosomal dominant, autosomal recessive, X-linked, or mitochondrial (maternal) inheritance. Metabolic disorders (e.g., Niemann-Pick type C, Tay-Sachs disease) may be “genetic” but present as late-onset ataxias without family history. [Nilantha de Silva, R. et al. Pract. Neurol. (2019) 19(3): 196-207].

[0034] Progressive cognitive disorders associated with the accumulation of neurofibrillary tangles ("NFTs") include, but are not limited to, progressive supranuclear palsy; dementia; dementia pugilistica; Creutzfeldt-Jakob disease; frontotemporal dementia; Pick's disease; other tau-positive pathological corticobasal degenerations; frontotemporal lobar degeneration (FTLD); and dementias lacking a unique histologic diagnosis.

[0035] Chronic Traumatic Encephalopathy (CTE) Chronic traumatic encephalopathy is considered a rare progressive degenerative disease of the brain seen in individuals with a history of repetitive head trauma. Chronic traumatic encephalopathy is still poorly understood because it can only be reliably diagnosed at postmortem autopsy. Originally called dementia pugilistica, it has been expanded to include brain injury from additional causes. Severe impacts received in cases of traumatic brain injury (TBI) can lead to CTE, especially if the impact is repeated. Repetitive head impacts (RHI), defined as exposure to repeated concussions and subconcussive events, have long been associated with CTE. [Nowinski, CJ et al. Front. Neurol. (2022) 13: 938163]. Even the relatively small impacts of mild traumatic brain injury (MTBI) or concussion may cause or contribute to the condition.

[0036] CTE is characterized by the progressive degeneration of brain tissue along with the accumulation of tau protein. The condition may begin within months of injury or may not begin until years or even decades after the last impact. Symptoms of CTE include behavioral changes, memory loss, depression, aggression, impulsive behavior, impaired judgment, and individuals gradually develop dementia. [www.protectthebrain.org, visited 2.6.2023]

[0037] Epilepsy Seizures are paroxysmal, cyclical changes in cortical electrical activity, almost always accompanied by behavioral changes. The changes may be purely subjective (i.e. sensory symptoms not evident to an outside observer), and sometimes such electrical activity may not produce any symptoms at all, the so-called subclinical or electrographic seizures. Focal or partial seizures originate from one hemisphere, the origin may be a small area or a large intrahemispheric multilobar network, and the discharge may then spread to diffusely involve both hemispheres. Generalized seizures originate from both hemispheres simultaneously. There are several different types of generalized seizures, the most common of which is the tonic-clonic ("grand mal") type, in which loss of consciousness with tonic stiffening is followed by a progressive increase in clonic (biphasic) muscle activity, all lasting about one minute. There are also isolated clonic and isolated tonic seizures, each characterized by its own form of motor activity without the other type, and the terrifying atonic seizures, in which the patient suddenly loses all muscle tone and falls to the ground, frequently causing self-harm. Commonly seen in children are absence ("petit mal") seizures, episodes of loss of consciousness and arrest of movement for a few seconds, sometimes accompanied by nystagmus, with the patient quickly returning to their previous activity as if nothing had happened (and usually unaware that anything had happened). Finally, myoclonic atonic seizures (sudden, momentary, lightning-like contractions of a muscle or group of muscles, accompanied by diffuse changes in the electroencephalography (EEG)), which are often not mentioned by the patient and usually have little impact on the patient's well-being, but can provide very important clues to properly classify the patient's epilepsy syndrome [Mintzer, S. Pharmacology and Therapeutics, Principles to Practice (2009), pages 663-83, Elsevier, Inc.]

[0038] Generalized epilepsy is further subdivided into idiopathic (primary) and symptomatic (secondary) where patients of the former have normal intelligence and brain function and the latter have generalized seizures secondary to some diffuse brain disease (e.g., congenital hypoxia, tuberous sclerosis) that also results in mental retardation.

[0039] Primary generalized epilepsy is genetic in nature, and it is widely believed that there are many specific genes associated with the various generalized epilepsy syndromes, only some of which have clearly identified functions. However, the inheritance and penetrance patterns appear to be complex, and the genes discovered so far account for only a small percentage of patients with primary generalized epilepsy. [Ibid., citing Berkovic, S. F. Genetics of epilepsy syndromes. In Engel, J. Pedley, T. (eds) Epilepsy: A comprehensive Textbook. Philadelphia: Lippincott-Raven (1997).

[0040] Focal epilepsy, at the “macro” level, can be caused by numerous focal lesions occurring anywhere in the cerebral cortex, including cortical dysplasias, tumors, heterotopias and hamartomas, congenital malformations, infarcts, hemorrhages, vascular malformations, traumatic injuries, and various types of central nervous system infections. The best-studied cause of focal epilepsy is hippocampal sclerosis, a condition with a very specific pattern of neuronal cell loss and gliosis distinct from that seen in anoxic injury, Alzheimer’s disease, or other hippocampal pathologies. 8 Hippocampal sclerosis is often associated with childhood neurological “hits” (e.g., prolonged febrile seizures, meningitis) and is likely the most common cause of drug-resistant focal epilepsy, often requiring resective surgery (discussed later in this chapter). Despite extensive research, and despite the notable success of hippocampal resection for this condition, it remains completely unclear what it is about this condition that generates seizures at the systems level.

[0041] At the "micro" level, seizures are conceptualized as resulting from excessive neural excitation, insufficient inhibition, or both. Focal seizures are the product of groups of electrically hyperexcitable neurons that fire synchronously and produce periodically evolving discharges. During a seizure, most such groups of neurons produce much more brief synchronous depolarizations, and in electric field recordings such as EEG, these appear as sharp-appearing waves (also called interictal epileptic activity, spikes, or sharp waves). These waves, and presumably the seizures arising from the same neurons, result from the summation of many paroxysmal depolarizing shifts in the postsynaptic neurons that are presumably due to excitatory postsynaptic potentials resulting from neurotransmitter release by the presynaptic neurons [Ibid., citing Prince, DA and Futamachi, KT. Intracellular recordings in chronic focal epilepsy. Brain Res. (1968) 11:681-4; Dichter, M. and Spencer, WA. Penicillin-induced interictal discharges from the cat hippocampus. I. Characteristics and topographical features. J. Neurophysiol. (1969) 32:649-62]. Recent evidence suggests that the excitatory neurotransmitters underlying many spontaneous depolarization shifts may originate from astrocytes rather than neurons [Ibid., citing Tian, ​​GF et al. An astrocytic basis of epilepsy. Nature Med. (2005) 11:973-81], but it is nonetheless clear that glutamate and its ionotropic receptors play a role in generating epileptic activity. Some anticonvulsants may act, at least in part, through glutamate (e.g., N-methyl-d-aspartate [NMDA] receptor antagonism), whereas others may act by reducing fast-frequency neuronal firing so that less glutamate is released at the synapse.In a similar manner, GABA, the major inhibitory neurotransmitter in the central nervous system, can apparently inhibit the occurrence of epileptic activity and reduce seizures, and some anticonvulsants (e.g., barbiturates, benzodiazepines, tiagabine) appear to exert their anticonvulsant properties by increasing synaptic GABA or enhancing its effects. It is worth noting that in some situations (e.g., in the absence of seizures as mentioned above), depending on the epileptogenic circuitry, neural hypersynchronization can occur through excessive rather than insufficient inhibition.

[0042] One nearly universal feature of seizures is that individual events end within minutes or even seconds. If the pathophysiological basis of seizure onset is largely unknown, the basis of seizure termination remains even more obscure. In some cases, seizures do not terminate, resulting in ongoing periodic cortical discharge, which has a striking clinical correlate, usually referred to as status epilepticus. It is presumed that seizure onset involves intrinsic seizure termination mechanisms in the brain, and status epilepticus results from the failure of these termination mechanisms. Because status epilepticus is a medical emergency under most circumstances, the pharmacological mechanisms involved in combating it are similar to those used for recurrent single seizures, although the delivery modes are generally more invasive (e.g., intravenous [IV], rectal, etc.).

[0043] Seizures can occur early (within the first week of brain injury) or late (more than one week after brain injury). Seizures that occur early after a traumatic brain injury are felt to be symptoms of the recent injury. Seizures that occur late after a TBI are more likely to recur and cause epilepsy.

[0044] According to the Epilepsy Foundation (www.epilepsy.com, visited 2.6.2023), approximately one in ten people (10%) will experience an early seizure after a TBI. 50% of early seizures occur within the first 24 hours after a TBI, and 25% of early seizures occur within the first hour after a TBI. Most very early seizures (within 24 hours of injury) are generalized tonic-clonic seizures.

[0045] Approximately 1 in 10 people develop status epilepticus early after a TBI. Young children are at highest risk for early post-traumatic seizures and status epilepticus.

[0046] People with more severe head injuries (e.g., car accidents, falls from a height, injuries from military explosions), brain swelling or blood outside the brain (subdural hemorrhage), or the brain being penetrated by a foreign object (e.g., bullet, combat injury), or prolonged loss of consciousness (more than 30 minutes) are more likely to have premature seizures. One in four people who suffer from bleeding in the brain that requires surgery (intracerebral hematoma) or a skull fracture that compresses or damages brain tissue will experience a premature seizure.

[0047] In some cases, even in people with "mild" head injuries and no evidence of brain damage on CT or MRI brain imaging, seizures may still occur.

[0048] EEG changes may or may not be present in the period immediately following a head injury. If there are markers of seizures on the EEG early after a TBI, this may mean that the person is more likely to develop epilepsy. People who have early seizures after brain trauma are at higher risk of developing post-traumatic epilepsy.

[0049] Seizures occurring more than one week after a traumatic brain injury are considered delayed seizures. Delayed seizures often occur because there is more severe damage to brain cells and the chemical environment around the cells has also changed. Delayed seizures are more likely to cause post-traumatic epilepsy complications.

[0050] People with post-traumatic epilepsy (PTE) are at risk for recurrent seizures as a result of brain injury. Approximately 1 in 50 people with traumatic brain injury will develop PTE. There is a spectrum of severity of PTE ranging from well-controlled seizures to highly disabling seizures that are resistant to treatment. Most seizures in post-traumatic epilepsy (8 in 10 people) are focal seizures that can spread to become bilateral tonic-clonic seizures.

[0051] One in two people develop traumatic epilepsy within one year of a brain injury. The more severe the head injury, the longer the risk of developing epilepsy. The risk of developing PTE decreases substantially over time, but for people with the most severe brain injuries, it can be as long as 15 years after the original traumatic injury. A person is at higher risk of developing PTE if they:

[0052] experiencing early seizures after TBI

[0053] Had bleeding into the brain tissue or a brain contusion (bruise) at the time of the injury,

[0054] A skull fracture occurs when part of the skull moves into the brain, damaging brain tissue or compressing the brain (depressed skull fracture),

[0055] A penetrating traumatic brain injury occurred (bullet, combat injury, etc.),

[0056] have a head injury related to alcohol use;

[0057] If there is trauma-related brain swelling, surgery was needed to remove bleeding from the brain (hematoma), to remove foreign objects from the brain, or to drain fluid from the brain (ventriculostomy),

[0058] EEG abnormalities appear early after injury.

[0059] People over 65 years of age, and / or

[0060] People with a family history of epilepsy.

[0061] Traumatic brain injury Traumatic brain injury (TBI) is a non-degenerative, non-congenital insult to the brain from an external mechanical force, possibly resulting in permanent or temporary impairment of cognitive, physical, and psychosocial functioning, with associated reduced or altered states of consciousness. Traumatic brain injury may manifest clinically from concussion to coma and death. TBI as a result of sports and recreational head injuries is becoming an increasing concern.

[0062] As depicted diagrammatically in Figure 1, TBI is divided into two subcategories: (1) primary injury, which occurs at the moment of trauma, and (2) secondary injury, which occurs immediately after the trauma and causes effects that may last for a long time. Secondary injuries may develop hours or days after the initial traumatic insult.

[0063] The primary injury may appear as a focal injury (e.g., skull fracture, intracranial hematoma, laceration, contusion, penetrating wound) or may be diffuse (such as diffuse axonal injury), which is a microscopic change that does not show up on a CT scan and is scattered throughout the brain.

[0064] Concussion is considered a mild form of diffuse axonal injury, which is caused by distortion of deep brain structures, leading to widespread neurological dysfunction that can cause impaired consciousness or coma.

[0065] One of the most widely used systems for classifying outcome from head injury is the Glasgow Outcome Scale (GOS).

[0066] [Table 1] 1Teasdale, G. and Jennett, B. Lancet (1974) 81-84; Teasdale, G. and Jennett, B. Acta Neurochir. (1976) 34:45-55.

[0067] For patients with closed head injuries, those with mild head injuries (usually defined as a GCS score at presentation of 13-15) tend to be successful. They may experience headaches, dizziness, irritability, or similar symptoms, but in most cases these improve gradually. Patients with moderate head injuries do not fare as well. Approximately 60% achieve a positive recovery, and an estimated 25% are left with moderate disability. Death or persistent vegetative state is the outcome in approximately 7-10% of cases. The remaining patients will have severe disability.

[0068] Despite its usefulness, the GOS is not a good tool for measuring subtle emotional or cognitive problems.

[0069] Other tools are available to evaluate athletes for concussion.

[0070] For example, the Sports Concussion Assessment Tool (SCAT5) is a standardized tool for use by physicians and licensed medical personnel to evaluate athletes age 13 and older for concussion. For children age 12 and younger, the Child SCAT5 is recommended.

[0071] The BESS test (Balance Error Scoring System) is a balance assessment protocol developed specifically to assess sports concussions. It is part of the Sports Concussion Assessment Tool (SCAT) developed by the Concussion in Sport Group as a standard protocol for concussion testing. It includes the interpretation of the Romberg test, which consists of a balance test first introduced in 1853. The BESS test is generally accepted by the clinical research community, but some concerns have been raised about the inter-rater and intra-rater reliability of manual scoring. Pressure measurement mat systems such as Tekscan's MobileMat™, combined with SportsAT™ software, aim to minimize the opportunity for human error by automating BESS test scoring. The MobileMat serves as a firm surface for the test subject to stand on, and a foam pad can be placed on the mat for that portion of the test. The mat is 0.3 inches thick and contains a high-resolution pressure sensor, adding objective data to the BESS test.

[0072] ImPACT Applications, Inc. offers a suite of concussion assessment tools, including a computerized battery of cognitive tests to aid in the management of concussions. For example, ImPACT Version 4 is applicable to individuals aged 12-80 years.

[0073] The King-Devick test is a two-minute rapid counting assessment in which an individual rapidly recites single-digit numbers and assesses impairments in eye movements, attention and language function.

[0074] Changes in cerebral glucose metabolism immediately following TBI 2A and 2B illustrate normal glucose metabolism (FIG. 2A) and the impairment of glycolytic pathways and oxidative metabolism of glucose immediately following TBI (FIG. 2B).

[0075] Upon impact, the rapid movement of the brain within the skull initiates a series of neurochemical perturbations that alter brain metabolism. Within minutes of injury, the ionic balance across neuronal membranes is disrupted, resulting in increased concentrations of extracellular potassium and glutamate, as well as intracellular calcium accumulation, depending on the severity of the injury [Prins, ML & Matsumoto, JH. The collective therapeutic potential of cerebral ketone metabolism in traumatic brain injury. J. Lipid Res. (2014) 55(12): 2450-7, Fineman I., et al. Concussive brain injury is associated with a prolonged accumulation of calcium: a 45Ca autoradiographic study. Brain Res. (1993) 624: 94-102; Katayama Y., et al. Massive increases in extracellular potassium and the indiscriminate release of glutamate following concussive brain injury. J. Neurosurg. (1999) 73: 889-900].This disruption of ionic balance requires cellular energy to re-establish homeostasis and is reflected by the increase in cerebral glucose uptake observed within 30 min after fluid percussion (FP) injury in adult rodents [ibid., citing Yoshino A., et al. Dynamic changes in local cerebral glucose utilization following cerebral conclusion in rats: evidence of a hyper-and subsequent hypometabolic state. Brain Res. (1991) 561: 106-119] and within 8 days after human TBI [ibid., citing Bergsneider M., et al. 1997. Cerebral hyperglycolysis following severe traumatic brain injury in humans: a positron emission tomography study. J. Neurosurg. (1997) 86: 241-251]. This transient increase in glucose uptake, also known as "hyperglycolysis," is followed by a prolonged period of decreased glucose metabolism. These cerebral metabolic changes are characteristic responses described in both experimental and clinical brain trauma.Experimental studies have shown that the magnitude and duration of glucose metabolic decline increases with injury severity and age [Ibid., Prins, M. et al. Mapping cerebral glucose metabolism during spatial learning: interactions of development and traumatic brain injury. J. Neurotrauma. (2001) 18:31-46; Thomas, S. et al. Cerebral metabolic response to traumatic brain injury sustained early in development: a 2-deoxy-D-glucose autoradiographic study. J. Neurotrauma. (2000) 17:649-665; Hovda, DA et al. Long-term changes in metabolic rates for glucose following mild, moderate, and severe concussive head injuries in adult rats. Soc. Neurosci. Abstract (1994). 20:845.

[0076] Other biochemical changes that occur immediately after TBI further disrupt glucose uptake and metabolism. 13Proton NMR spectroscopy of [C]-labeled glucose shows a 9-12% increase in glucose disposal through the pentose phosphate pathway 3-24 hours after controlled cortical impact (CCI) injury, thereby decreasing the available glucose supply for energy production [Ibid., citing Bitnik, BL et al. Upregulation of pentose phosphate pathway and preservation of tricarboxylic acid cycle flux after experimental brain injury. J. Neurotrauma (2005). 22:1052-1065].TBI has also been shown to generate an early increase in reactive oxygen species (ROS) that damage lipids, proteins, and DNA [Ibid., Hall, ED et al. Brain hydroxyl radical generation in acute experimental head injury. J. Neurochem. (1993) 60:588-594; Althaus, JS et al. The use of salicylate hydroxylation to detect hydroxyl radical generation in ischemic and traumatic brain injury. Reversal by tirilazad mesylate (U-74006F). Mol. Chem. Neuropathol. (1993) 20:147-162; Marklund, N. et al. Effects of the nitrone radical scavengers PBN and S-PBN on in vivo trapping of reactive oxygen species after traumatic brain injury in rats. J. Cereb. Blood Flow Metab. (2001) 21:1259-1267; Sen, S. et al. al. Oxypurinol inhibits free radical release from the cerebral cortex of closed head injured rats. Neurosci. Lett. (1993) 162: 117-120; Sens, S. et al. Alpha-phenyl-tert-butyl-nitrone inhibits free radical release in brain concussion. Free Radic. Biol. Med (1994). 16: 685-691]. ROS-induced DNA damage activates DNA repair enzymes such as poly-ADP ribose polymerase (PARP). In the presence of DNA strand breaks, pathological activation of PARP increases the cytoplasmic NAD. +This ultimately inhibits glycolytic processing of glucose at the glyceraldehyde phosphate dehydrogenase step [Ibid., citing Sheline, C. et al. Zinc-induced cortical neuronal death: contribution of energy failure attributable to loss of NAD(+) and inhibition of glycolysis. J. Neurosci. (2000) 20:3139-3146. Collectively, this series of biochemical changes redirects and impedes the processing of glucose through the glycolytic pathway. As a result, glucose oxidation and ATP concentrations are reduced [Ibid., citing Singh, IN et al. Time course of post-traumatic mitochondrial oxidative damage and dysfunction in a mouse model of focal traumatic brain injury: implications for neuroprotective therapy. J. Cereb. Blood Flow Metab. (2006) 26:1407-1418; Lee, S. et al. Evidence for energy failure following irreversible traumatic brain injury. Ann. NY Acad. Sci. (1999) 893:337-340], making glucose an inefficient energy substrate in the brain after TBI.

[0077] TBI induces an inflammatory response in the brain Injury to the CNS also induces an inflammatory response from resident microglia and macrophages, as well as peripheral immune cells such as neutrophils, monocytes, and T cells. Microglia and resident macrophages respond immediately to injury after sensing the presence of damage-associated molecular patterns (DAMPs), such as adenosine triphosphate (ATP) or intracellular proteins released from damaged or dying cells. Signaling from DAMP receptors initiates local cytokine and chemokine production, which influences the surrounding environment and provides clues for peripheral immune infiltration (Russo, M. and McGovern, DB, “Inflammatory neuroprotection following traumatic brain injury”, Science (2016) 353(6301):783-6, citing Corps, KN. Et al. JAMA Neurol. (2015) 72:355-62).

[0078] Within the CNS, positive aspects of immune cells recruited in response to injury include removing dead cells, supporting the barrier system, and setting the stage for wound healing. Microglia, monocytes, macrophages, neutrophils, and T cells can collectively orchestrate responses that preserve neural tissue and promote regeneration.

[0079] In general, microglia are key participants in the acute phase of CNS injury. Seal the barrier and clear debris are just two benefits of involving these cells in the response. Due to their ubiquitous presence and abundance of DAMP sensors, microglia are usually one of the first responders to brain injury. For example, microglia express several purinergic receptors that allow them to respond rapidly to extracellular purines such as ATP. Upon detection of extracellular ATP, microglia direct their processes towards the site of injury within minutes. Microglia also participate in the support of the immediate CNS barrier after injury to brain capillaries (ibid. citing Lou, N. et al. Proc. Natl Acad. Sci. USA (2016) 113:1074-9). In addition to supporting the barrier, microglia and macrophages can also clear debris from the injured CNS.

[0080] Despite the various protective layers that separate the CNS from peripheral tissues, immune cells infiltrate the CNS in response to trauma. In a model of spinal cord injury, neutrophil recruitment was necessary for proper wound healing and repair (Id., citing Stirling, DP et al. J. Neurosci. (2009) 29:753-64). Mice treated with neutrophil-depleting antibodies showed reduced astrocytic reactivity at the injury site, larger lesions, and worse neurological outcomes. Neutrophils were also recruited to the meninges after acute brain injury in a purinergic receptor-dependent manner, and interference with this response increased the amount of meningeal cell death (Id., citing Roth, TL et al. Nature (2014) 505:223-8). It is currently unclear how neutrophils contribute to the neuroprotective response after CNS injury, but these cells have the ability to recruit peripheral monocytes within a day or two. Spinal cord injury induces a marked recruitment of pro-inflammatory macrophages, followed several days later by wound-healing macrophages (Id., citing Shechter, R. et al. Immunity (2013) 38:555-69). When this wound-healing response was blocked, mice were unable to properly repair the lesion and regain motor skills (Id., citing Shechter, R. et al. Immunity (2013) 38:555-69). As in peripheral tissues, innate immune cells are essential for tissue remodeling and repair in the CNS.

[0081] T cells can also play an active role in the CNS injury response. CNS injury can promote the nonspecific recruitment of CD4+ T cells that produce interleukin-4 (IL-4) in a major histocompatibility complex II-independent manner (Id., citing Walsh, JT et al. J. Clin. Invest. (2015) 125:699-714). Release of IL-4 enhances neurotrophin signaling that helps stimulate axonal regrowth after injury. Mice lacking T cells or IL-4 showed increased neuronal loss and neuronal dysfunction after CNS injury. During CNS tissue repair, effector and regulatory CD4+ T cells function in tandem. Regulatory T cells (Tregs) often keep the immune response in check by modulating inflammatory mediators to promote wound healing and remodeling. Depletion of Tregs prior to CNS injury increased the recruitment of effector CD4+ T cells and improved neurological recovery (Id. citing Raposo, C. et al. J. Neurosci. (2014) 34:10141-55). In contrast, depletion of Tregs several days after injury disrupted the tissue repair process. Similar to the transition from proinflammatory to wound-healing macrophages, these Treg data indicate that over time, the neuroinflammatory response to injury changes.

[0082] Currently, no treatments are available that prevent neuronal damage or promote neurohealing after TBI. Attempts to stop neuroinflammation have been successful in treating multiple sclerosis but have failed in other conditions, because anti-inflammatory strategies block the beneficial as well as the harmful aspects of inflammation.

[0083] Metabolic Relationship of the Ketogenic Diet and Beta-Hydroxybutyrate (BHB) to Cellular Metabolism and Energy Metabolism is the sum of the chemical and physical changes that occur within a tissue. It consists of anabolism, the reactions that convert small molecules into larger molecules, and catabolism, the reactions that convert larger molecules into smaller molecules.

[0084] catabolism Under normal circumstances, the proteins, lipids and polysaccharides that make up the majority of the food we eat must be broken down into smaller molecules before our cells can use them. The enzymatic breakdown of these molecules (i.e., catabolism) proceeds in three stages. In the first stage, the large polymer molecules are broken down by digestion into their monomeric subunits (proteins into amino acids, polysaccharides into sugars, and fats into fatty acids and glycerol). These processes occur outside the cell, mainly by the action of secreted enzymes. In the second stage, the resulting small molecules enter the cell and are further broken down in the cytoplasm. Most of the carbon and hydrogen atoms of the sugars are converted into pyruvate, which then enters the mitochondria, where it is converted into the acetyl group of acetyl coenzyme A (acetyl CoA). Acetyl CoA, like ATP, is a chemically reactive compound that releases large amounts of energy when hydrolyzed. Large amounts of acetyl CoA are also produced by the oxidation of fatty acids. The third stage occurs when the acetyl group of acetyl CoA is completely broken down into CO2 and H2O. Most of the ATP is produced in this final stage. With the generation of ATP, the energy originally derived from the combustion of carbohydrates and fats is redistributed as chemical energy in a conveniently packaged form that is easily released. [Alberts, B. et al. Molecular Biology of the Cell (1983) Garland Publishing, NY, Ch. 2, pages 67-75].

[0085] The most important part of the second stage of catabolism is glycolysis, which refers to a series of reactions involving the splitting of glucose. In glycolysis, a glucose molecule with six carbon atoms is converted into two pyruvate molecules, each with three carbon atoms, by a series of nine enzymatic reactions involving a series of phosphate-containing intermediates. At the end of glycolysis, the ATP balance sheet shows a net gain of two molecules of ATP per glucose molecule. In most animal cells, the pyruvate formed immediately enters the mitochondria and is completely oxidized to CO2 and water.

[0086] In addition to pyruvate, fatty acids and some amino acids also pass from the cytosol to the mitochondria, where they are converted to acetyl-CoA or one of the other intermediates of the citric acid cycle.

[0087] Glucose homeostasis Glucose, the fundamental source of cellular energy, is released by the breakdown of endogenous glycogen stores located primarily in the liver. Glucose is also released indirectly in muscle via intermediate metabolic products. These whole-body energy stores are replenished from dietary glucose, which is digested, absorbed across the intestinal wall, and then distributed to various tissues of the body. (Bryant, NJ et al, Nat. Rev. Mol. Cell Biol. 2002:3(4):267-77).

[0088] Normally, after glucose ingestion, an increase in plasma glucose concentration triggers insulin release, which stimulates visceral (liver and gastrointestinal tissues) and peripheral glucose uptake and suppresses endogenous (mainly hepatic) glucose production. In healthy adults, blood glucose levels are tightly regulated within the range of 70-99 mg / dL to meet metabolic requirements and are maintained by specific hormones (e.g., insulin, glucagon, incretins) and the central and peripheral nervous systems. Various cells and tissues (in the brain, muscle, gastrointestinal tract, liver, kidney and adipose tissue) also participate in blood glucose regulation by uptake, metabolism, storage and secretion. DeFronzo, RA., Med. Clin. N. Am. 88:787-835 (2004); Textbook of Medical Physiology; Gerich, JE, Diabetes Obes. Metab. 2000:2:345-350. Under normal physiological circumstances, glucose levels rarely exceed 140 mg / dL, even after ingesting a high-carbohydrate meal.

[0089] Insulin is a potent antilipolytic (inhibits fat breakdown) hormone that is known to lower blood glucose by accelerating the transport of fructose into insulin-sensitive cells and promoting its conversion to storage compounds via glycogenogenesis (conversion of glucose to glycogen) and lipogenesis (fat formation). Glucagon, which also plays a role in glucose homeostasis, is produced in response to a decline in normal glucose levels or hypoglycemia and acts to increase glucose levels by accelerating glycolysis and promoting gluconeogenesis. After a glucose-containing meal, glucagon secretion is inhibited by hyperinsulinemia, which contributes to the suppression of hepatic glucose production and the maintenance of normal postprandial glucose tolerance.

[0090] Incretins, which include glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1), are also involved in regulating blood glucose, in part through their effects on insulin and glucagon. (Drucker, and Nauk, Lancet 368:1696-1705 (2006). However, both GLP-1 and GIP are considered glucose-dependent hormones, meaning that they are secreted only when glucose levels are above normal fasting plasma glucose levels. Normally, these hormones are released in response to a meal and help stimulate insulin secretion by activating specific receptors on pancreatic beta cells. However, when glucose levels are low, GLP-1 and GIP levels (and their stimulatory effect on insulin secretion) are reduced. Drucker, Cell Metab. 3:153-165 (2006).

[0091] Ketosis As an alternative to glucose utilization, the body can metabolically move into a state of ketosis. Ketosis relies on fat-derived ketones produced in the liver to fuel nearly every cell in the body. Ketones - acetoacetate, β-hydroxybutyrate, and acetone - are water-soluble molecules produced by the liver from fatty acids when blood glucose and hepatic glycogen stores are minimized. Glycogen depletion occurs and ketone levels rise during fasting, low carbohydrate intake, intense exercise, periods of starvation, or due to complete lack of insulin in untreated type I diabetes. [Gershuni, VM et al. Curr. Nutr. Rep. (2018) 7 (3): 97-106].

[0092] Hormonal activation of lipolysis and ketogenesis is mediated by epinephrine and glucagon and is blocked by insulin. Low carbohydrate diets result in low insulin and increased glucagon. In addition to stimulating glycolysis in the liver, glucagon stimulates lipolysis to release stored fatty acids from adipose tissue. [Ibid., citing Sato, K. et al. Insulin, Ketone bodies and mitochondrial energy transduction. FASEB J. (1995) 9:651-58]. In addition to forming ketones, fatty acids can be converted to acetyl-CoA, an intermediate substrate between fatty acid oxidation and glucose metabolism, enter the citric acid cycle, and then undergo oxidative phosphorylation for ATP generation. Conversely, in response to high blood glucose (i.e., after a high-carbohydrate meal), insulin levels rise, shutting off ketogenesis in favor of de novo lipogenesis (fat storage). Ketosis therefore represents a shift away from an insulin-mediated glucose-dependent state towards an increased ability to use dietary fat and fat stores for fuel.

[0093] Nutritional Ketosis Nutritional ketosis is the intentional restriction of dietary carbohydrate intake to induce metabolic effects that accelerate ketone production, stabilize blood glucose, and minimize insulin release, thereby mitigating the downstream anabolic and tumorigenic effects of long-standing insulin resistance. As described by Volek and Phinney, a "well-formulated" ketogenic diet consists of 5-10% carbohydrate (<20-50 g / day), sufficient protein (1-1.5 g / kg / day), and fat to satiety. Nutritional ketosis is characterized by blood ketone levels of 0.5-3 mg / dL [ibid., citing Volek, JS and Phinney SD. The Art and Science of Low Carbohydrate Living (Beyond Obesity, LLC, Miami FL USA)]. This is in stark contrast to, and should not be confused with, the pathophysiological state of type 1 diabetic ketoacidosis (DKA). Despite the similar names, they are two distinct metabolic processes. Endogenous insulin production is protective against the development of DKA, and the range of ketones present in DKA is 5-10 times greater than the levels achieved during nutritional ketosis. In addition, in nutritional ketosis, the body is able to maintain normal blood glucose levels and maintain a normal pH, as opposed to the very high blood sugar and acidic pH associated with DKA.

[0094] After a few weeks, a "keto-adapted state" occurs, which refers to the body's ability to adapt and respond to using primarily ketones for fuel. One potential reason this occurs is secondary to the upregulation of transcription of genes encoding metabolic machinery that leads to increased mitochondrial density in oxidative tissues such as the brain and muscle. Mouse studies suggest that this may occur through increased mitogenesis or reduced mitochondrial damage. [Ibid., citing Bough, KJ et al. Mitochondrial biogenesis in the anticonvulsant mechanism of the ketogenic diet. Ann. Neurol. (2006) 60:223-35; Ahola-Erkkila, S. et al. Ketogenic diet slows down mitochondrial myopathy progression in mice. Hum. Mol. Genet. (2010) 19:1974-84] In addition, ketones can induce epigenetic regulation via histone deacetylation inhibition, indicating that they are signaling molecules in addition to being an energy source. [Ibid. citing Newman, JC and Verldin E. Beta-hydroxybutyrate: much more than a metabolite. Diabetes Res. Clin. Pract. 2014 106:173-81].

[0095] Through this process, fat-derived energy is generated in the liver and then transported throughout the body to fuel the brain, renal cortex, heart, and skeletal muscles. [Ibid., citing Laffel, L. Ketone Bodies: a review of physiology, pathophysiology and application of monitoring to diabetes. Diabetes Metab. Res. Rev. (1999) 15:412-26]. Ketones can provide up to 60% of the ATP required by the body [Ibid., citing Veech, RL et al. Ketone bodies, potential therapeutic uses. IUBMB Life (2001) 51:241-47], the remainder being derived from endogenous gluconeogenesis, which utilizes glycerol forms of triglycerides and glycogen amino acids from proteins for glucose production. Ketones cross the blood-brain barrier and replace glucose as the brain's primary energy source. Pioneering work by George Cahill using a model of starvation ketosis revealed that the brain has metabolic flexibility and can switch from being a glucose-dependent organ (approximately 150 g / day) to one that derives more than two-thirds of its energy from ketones [Ibid. citing Veech, RL et al. Ketone bodies, potential therapeutic uses. IUBMB Life (2001) 51:241-47; Westman, EC et al. Low-carbohydrate nutrition and metabolism. Am. J. Clin. Nutr. (2007) 86:276-84].The main features of the ketogenic diet are the establishment of ketosis and stabilization of insulin levels, which address biomarkers of metabolic syndrome. [Ibid., Volek, JS et al. Carbohydrate restriction has a more favorable impact on the metabolic syndrome than a low-fat diet. Lipids (2009) 44:297-309; Volek, JS & Feinman, RD. Carbohydrate restriction improves the features of Metabolic syndrome. Metabolic Syndrome may be defined by the response to carbohydrate restriction. Nutr. Metab. (Lond) (2005) 2:31l]. By lowering insulin rise, lipids are released from stores and oxidized. [Ibid., Volek JS, et al. Dietary carbohydrate restriction induces a unique metabolic state positively affecting atherogenic dyslipidemia, fatty acid partitioning, and metabolic syndrome. Prog Lipid Res (2008) 47, 307-318.

[0096] However, intravenous ketone administration may be an inefficient method to induce changes in brain energy metabolism [Prins, ML and Matsumoto, JH. J. Lipid Res. (2014) 55: 2450-55, citing Pan JW, et al (2001). Measurement of beta-hydroxybutyrate in acute hyperketonemia in human brain. J. Neurochem. 79: 539-544; White, H. et al. Effect of a hypertonic balanced ketone solution on plasma, CSF and brain beta-hydroxybutyrate levels and acid-base status. Intensive Care Med. (2013) 39: 727-733; Pan, J. et al. Human brain beta-hydroxybutyrate and lactate increase in fasting-induced ketosis. J. Cereb. Blood Flow Metab. (2000) 20: 1502-1507]. Strategies designed to primarily increase plasma ketone levels must also target increased ketone transport across the blood-brain barrier (BBB) ​​by monocarboxylate transporters (MCTs), membrane proteins that act as carriers of lactate, pyruvate, and ketone bodies. Furthermore, the high doses required to achieve desired ketone levels can cause severe side effects involving insulin / glucagon balance.[Id., Clarke,:K.et al.Kinetics,safety and tolerability of(R)-3-hydroxybutyl(R)-3-hydroxybutyrate in healthy adult subjects.Regul.Toxicol.Pharmacol.(2012)63:401-408;van Delft,R.et al.Blood beta-hydroxybutyrate correlates better with seizure reduction due to ketogenic diet than do ketones in the cite urine.Seizure.(2010)19:36-39;Madison,LL,et al.The hypoglycemic action of ketones.II.Evidence for a stimulatory feedback of ketones on the pancreatic beta cells.J.Clin.Invest.(1964)43:408-415].

[0097] HCA2 and its role in protection against ischemic brain injury HCA2, originally named GPR109A, is a G protein-coupled receptor that belongs to the hydroxyl carboxylic acid (HCA) receptor family. [Offermanns, S. and Schwaninger, M. Nutritional or pharmacological activation of HCA2 ameliorates neuroinflammation. Trends in Molec. Med. (2015) 21(40): 345-55, citing Offermanns, S. et al. (2011) Pharmacol. Rev. (2011) 63: 269-90]. It is linked to the G family of G proteins [ibid., Soga, T. et al. Molecular identification of nicotinic acid receptor. Biochem. Biophys. Res. Commun. (2003) 303: 364-69; Tunaru, S. et al. PUMA-G and HM74 are receptors for nicotinic acid and mediate its anti-lipolytic effect. Nat. Med. (2003) (9): 352-55; Wise, A. et al. Molecular identification of high and low affinity receptors for nicotinic acid. J. Biol. Chem. (2003) 278: 9869-74; Taggart, AK et al. D-β-hydroxybutyrate inhibits adipocyte lipolysis via the nicotinic aid receptor PUMA-GJ Biol. Chem. (2005) 280:26649-52; citing Benyo, Z. et al. GPR109A (PUMA-G / HM74A) mediates nicotinic acid-induced flushing. J. Clin. Invest. (2005) 115:3634-40], is activated by the ketone body BHB, and to a lesser extent by butyrate [ibid., Taggart, AK et al.D-β-hydroxybutyrate inhibits adipocyte lipolysis via the nicotinic aid receptor PUMA-GJ Biol.Chem.(2005)280:26649-52]. HCA2 is also expressed in various immune cells, including neutrophils, macrophages, epidermal Langerhans cells, dendritic cells, and microglia, but not in lymphocytes [ibid., Benyo, Z. et al. GPR109A (PUMA-G / HM74A) mediates nicotinic acid-induced flushing. J. Clin. Invest. (2005) 115: 3634-40; Kostylinn, G. et al. Neurotrophil apoptosis mediated by nicotinic acid receptors (GPR109A). Cell Death Differ. (2005) 15: 134-42; Kostylina, G. et al. Neurotrophil apoptosis mediated by nicotinic acid receptors (GPR109A). Cell Death Differ. (2008) 15: 134-42; Maxiejewski-Lenoir, D. et al. al.Langerhans cells release prostaglandin D2 in response to nicotinic acid.J.Invest.Dermatol.(2006)126:2637-46;Rahman,M.et al.The β-hydroxybutyrate receptor HCA2 activates a neuroprotective subset of macrophages.Nat.Commun.(2014)5:3944;Schaub,A.et al.PUMA-G,an IFN-γ-inducible gene in macrophages is a novel member of the seven transmembrane spanning receptor superfamily.Eur.J.Immunol.(2001)31:3714-25;Singh,N.et al.Activation of Gpr109a, receptor for niacin and the commensal metabolite butyrate, suppresses colonic inflammation and carcinogenesis. Immunity (2014) 40:128-39]. BHB is the main endogenous ligand for HCA2 and activates human HCA2 with an EC50 of approximately 700 μM [ibid., citing Taggart, AK et al. D-β-hydroxybutyrate inhibits adipocyte lipolysis via the nicotinic aid receptor PUMA-GJBiol. Chem. (2005) 280:26649-52], a concentration that this ketone body reaches only after fasting or on a ketogenic diet. HCA2 can also mediate anti-inflammatory and immunomodulatory effects through its expression by immune cells, and it has been suggested that this activity, when induced by ketone bodies during metabolism, helps to conserve energy. [Ibid., citing Lukasova, M. et al. Nicotinic acid (niacin): new lipid-independent mechanisms of action and therapeutic potentials. Trends Pharmacol. Sci. (2011) 32: 700-7].

[0098] Recent evidence indicates that HCA2 can mediate the neuroprotective effects of BHB [Ibid. citing Rahman, M. et al. The β-hydroxybutyrate receptor HCA2 activates a neuroprotective subset of macrophages. Nat. Commun. (2014) 5:3944]. The ketogenic diet is well established as a treatment for pediatric epilepsy [Id., citing Levy, RG et al. Ketogenic diet and other dietary treatments for epilepsy. Cochrane Database Syt. Rev. (20120 3:CD001903)], and preclinical studies have provided evidence that the ketogenic diet may also be protective in experimental models of other neurological disorders, including TBI. Research into the therapeutic potential of the ketogenic diet is driven by the concept of improving energy balance in affected brains. However, BHB is not only a metabolic intermediate but also a signaling molecule that activates HCA2 and inhibits GPR41 and histone acetylases (HDACs) 1, 3, and 4 [Id., Newman, JC and Verdin, E. Ketone bodies as signaling metabolites. Trends Endocrinol. Metab. (2014) 25:42-52; Won, YJ et al. β-hydroxybutyrate modulates N type calcium channels in rats. sympathetic neurons by acting as an agonist for the G protein coupled receptor FFAS. J. Neurosci. (2013) 33:19314-25], which led to the hypothesis that BHB induces a neuroprotective effect through a specific action on a receptor or enzyme.A recent study of the ketogenic diet in a mouse model of stroke supported the role of HCA2 in the BHB effect in this disease model [Ibid., citing Rahman, M. et al. The β-hydroxybutyrate receptor HCA2 activates a neuroprotective subset of macrophages. Nat. Commun. (2014) 5:3944]. These researchers concluded that additional supply of energy substrates is not sufficient to explain the protection offered by BHB, but rather activation of HCA2 on monocyte-derived cells, neutrophils or microglia is necessary to mitigate brain damage. Without being limited by theory, HCA2 activation may be a therapeutic principle for treating brain disorders with a neuroinflammatory component. For example, HCA2 on monocyte-derived cells may be involved in the upregulation of neuroprotective Ly6C in the brain. lo It has been hypothesized that Ly6C may provide a target for stimulating monocyte-derived cells. Lymphocyte antigen 6 complex (Ly6C) is a marker for macrophage subsets. hi Monocytes have proinflammatory and antibacterial functions and express high levels of CC chemokine receptor 2 (CCR2) and low levels of CX3C chemokine receptor 1 (CX3CR1) [Kratofil,RM et al.“Monocyte Conversion During Inflammation and Injury.Arteriosclerosis, Thrombosis and Vascular Biology(2017)37(1):35-42, citing Geissmann,F.et al.Blood monocytes consist of two principal subsets with distinct migratory properties.Immunity(2003)19:71-82].Patrol monocytes, also known as Ly6C low Monocytes survey the vasculature by constantly migrating along its lumen and are involved in the early response to inflammation and tissue repair.

[0099] Figure 3 illustrates the direct and indirect signaling effects of BHB in a schematic way. In addition to the direct signaling effects of BHB, indirect signaling mechanisms enhance the influence of BHB through downstream effects seen on molecules that have been removed at least once. In contrast to direct signaling functions attributable to the BHB compound itself, indirect signaling functions require catabolism to other molecules, Abbreviations: BHB, β-hydroxybutyrate; CoA, coenzyme A; FFAR3, free fatty acid receptor 3; GABA, γ-amino-butyric acid; HDAC, histone deacetylase; HCAR2, hydroxycarboxylic acid receptor 2; NAD, nicotinamide adenine dinucleotide; VGLUT, vesicular glutamate transporter. [Quoted from Newman JC and Verdin, E. β-hydroxybutyrate: a signaling metabolite. Ann. Rev. Nutrition (2017) 37:51-76].

[0100] Beta-hydroxybutyrate (BHB) has a molecular weight of 104.1 g / mol. As a small polar molecule, BHB is readily soluble in water and blood. [Newman, JC and Verdin, E β-Hydroxybutyrate. Annu. Rev. Nutr. (2017) 37: 51-76]. BHB is a chiral molecule at the 3' hydroxyl group. There are two enantiomers, R / d and S / l, typically designated as D and L, respectively. Human liver produces the D enantiomer of the molecule. [Chriett, S. et al. Prominent action of butyrate over β-hydroxybutyrate as histone deactylase inhibitor, transcriptional modulator and anti-inflammatory molecule. Sci. Reports (2019) 9 (1): 742]. R-BHB is a normal product of human and mouse metabolism. S-BHB itself is not a normal product of human metabolism. In experiments involving the infusion of labeled R-BHB, S-BHB, or a mixture thereof into rats or pigs, S-BHB was found to be largely converted to R-BHB (Newman, JC and Verdin, E β-Hydroxybutyrate. Annu. Rev. Nutr. (2017) 37:51-76 citing Lincoln, BC et al. Arch. Biochem. Biophys. (1987) 259:149-56), and although the molecular pathway for this is not known, it may occur via conversion of S-BHB to acetyl-CoA, followed by production of R-BHB from acetyl-CoA. At least a portion of the S-BHB is also ultimately converted to CO2, likely after being metabolized to acetyl-CoA.Data show that at the same dose, the D enantiomer is superior in increasing D-beta-hydroxybutyrate levels when compared to a racemic mixture of the molecule [Cuenoud, B. et al. Metabolism of exogenous D-beta-hydroxybutyrate, an energy substrate avidly consumed by the heart and kidney. Frontiers Nutrition (2020) 7: 13]. Many of the currently available exogenous forms of beta-hydroxybutyrate are racemic mixtures, salt forms, or variations of the true molecule, such as 1,3-butanediol.

[0101] The data indicate that the effectiveness of D-beta hydroxybutyrate for neurological indications is limited by the rate at which D-beta hydroxybutyrate can be transported across the BBB to sufficiently increase D-beta hydroxybutyrate concentrations in the brain. [White, H. et al. A systematic review of intravenous β-hydroxybutyrate use in humans-a promising future therapy? Frontiers Medicine (2021) 8: 740374]. Several monocarboxylate transporters, including MCT1 and MCT2, carry BHB across the blood-brain barrier (Pellerin, L. et al. J. Neurosci. Res. (2005) 79: 55-64), and their expression can regulate brain BHB uptake (ibid. citing Barton, KM, and Palmer, SE (2016) Curr. HIV / AIDS Rep. (2016) 13: 77-84).

[0102] The present disclosure provides a targeted approach to deliver synthetic D-beta hydroxybutyrate to the brain, bypassing the BBB, where controlled release of beta-hydroxybutyrate can result in increased levels of D-beta hydroxybutyrate in the brain both immediately after administration and over time. [Brief description of the drawings]

[0103] [Figure 1] Depicts a cascade of negative effects in the brain of traumatic insults. Primary injury occurs within the first seconds after impact and is generally mechanical in nature. Secondary injury occurs within minutes to weeks after impact and affects the brain's ability to properly regulate ion balance, metabolism, and inflammation. Tertiary injury, or neuropathology, occurs weeks to years after impact and is thought to be caused by the energy crisis and oxidative stress of secondary injury. Daines, SA et al. The therapeutic potential and limitations of ketones in traumatic brain injury. Frontiers Neurology (2021), 12: 723148, Quoted from Figure 1.

[0104] [Figure 2A] Schematic comparison of glucose metabolism in normal (Figure 2A) and post-TBI (Figure 2B) physiology. Glucose metabolic pathways are impaired in six ways: glycolytic pathways and oxidative metabolism of glucose after TBI. (1) A transient increase in glucose uptake followed by a long-term decline in glucose metabolism; (2) More glucose-6-phosphate is diverted to the pentose phosphate pathway to generate protective and repair molecules; (3) Glyceraldehyde-3-phosphate dehydrogenase activity slows, resulting in a decline in tricarboxylic acid cycle intermediates; (4) Astrocyte lactate production increases, diverting lactate to energy-starved neurons; (5) Pruvate dehydrogenase complexes are inhibited, resulting in a decline in tricarboxylic acid cycle intermediates; (6) Oxidative phosphorylation leading to impaired electron transport chain function and reduced ATP production. Daines, SA et al. The therapeutic potential and limitations of ketones in traumatic brain injury. Frontiers Neurology (2021), 12: 723148, Quoted from Figure 2. [Figure 2B]Schematic comparison of glucose metabolism in normal (Figure 2A) and post-TBI (Figure 2B) physiology. Glucose metabolic pathways are impaired in six ways: glycolytic pathways and oxidative metabolism of glucose after TBI. (1) A transient increase in glucose uptake followed by a long-term decline in glucose metabolism; (2) More glucose-6-phosphate is diverted to the pentose phosphate pathway to generate protective and repair molecules; (3) Glyceraldehyde-3-phosphate dehydrogenase activity slows, resulting in a decline in tricarboxylic acid cycle intermediates; (4) Astrocyte lactate production increases, diverting lactate to energy-starved neurons; (5) Pruvate dehydrogenase complexes are inhibited, resulting in a decline in tricarboxylic acid cycle intermediates; (6) Oxidative phosphorylation leading to impaired electron transport chain function and reduced ATP production. Daines, SA et al. The therapeutic potential and limitations of ketones in traumatic brain injury. Frontiers Neurology (2021), 12: 723148, Quoted from Figure 2.

[0105] [Diagram 3] Schematic illustrating the direct and indirect signaling effects of BHB. Abbreviations: BHB, β-hydroxybutyrate; CoA, coenzyme A; FFAR3, free fatty acid receptor 3; GABA, γ-amino-butyric acid; HDAC, histone deacetylase; HCAR2, hydroxycarboxylic acid receptor 2; NAD, nicotinamide adenine dinucleotide; VGLUT, vesicular glutamate transporter. Quoted from Newman JC and Verdin, E. β-hydroxybutyrate: a signaling metabolite. Ann. Rev. Nutrition (2017) 37:51-76.

[0106] [Figure 4]This is an illustrated diagram showing the anatomical structure inside the nose in cross section [quoted from Janfaza, P. et al. Surgical anatomy of the head and neck., Harvard University Press (June 15, 2011), Chapter 5.

[0107] [Figure 5A] FIG. 1 is a graph of beta-hydroxybutyrate (BHB) concentration in plasma (μg / mL, y-axis) over time (in hours) (x-axis) on day 14 following intranasal administration to C57BL / 6J mice for Groups 1 (vehicle), 2 (30 mg / kg BHB salt), and 3 (BHB ester (15 mg / kg). [Figure 5B] FIG. 1 is a graph of beta-hydroxybutyrate (BHB) concentration in the brain (μg / mL, y-axis) over time (in hours) (x-axis) on day 14 following intranasal administration to C57BL / 6J mice for Groups 1 (vehicle), 2 (30 mg / kg BHB salt), and 3 (BHB ester (15 mg / kg).

[0108] [Figure 6A] FIG. 1 is a graph of beta-hydroxybutyrate (BHB) concentration in plasma (ug / g, y-axis) over time (in hours) (x-axis) on day 14 following intranasal administration to C57BL / 6J mice for Groups 1 (vehicle), 2 (30 mg / kg BHB salt), and 3 (BHB ester (15 mg / kg). [Figure 6B] FIG. 1 is a graph of beta-hydroxybutyrate (BHB) concentration in brain (ug / g, y-axis) over time (in hours) (x-axis) on day 14 following intranasal administration to C57BL / 6J mice for Group 1 (vehicle), Group 2 (30 mg / kg BHB salt), and Group 3 (BHB ester (15 mg / kg)).

[0109] [Figure 7]7 is a graph showing endogenous baseline levels of beta-hydroxybutyrate (BHB) (y-axis) over time (x-axis) when BHB was administered at 15 mg / kg and 30 mg / kg. Note that the endogenous baseline in FIG. 7 is the same as the vehicle line in FIG. 5A (approximately 19 μg / mL).

[0110] [Figure 8] Bar graphs of the results of the beam walking behavior assay after TBI or sham injury are shown. The number of footfalls is plotted against the sample test group of C57BL / 6J female mice, 20 animals per group. 20 μl of a standard stock solution (50 mg / kg) of the test compound R-(-)-3-hydroxybutyric acid sodium salt (Sigma,) (BHB), freshly prepared each day, was administered intranasally daily starting on day 0 immediately after TBI until day 14. Saline was added to the compound to the appropriate concentration. The test groups were: (1) sham injury, vehicle treated; (2) TBI injury, vehicle treated; (3) pre-treated with BHB IN before TBI injury, then treated with saline after TBI injury; (4) pre-treated with saline before TBI injury, then treated with BHB after TBI injury; (5) pre-treated with BHB IN before TBI injury, then treated with BHB after TBI injury, and pre-treated with 2x dose of BHB IN before TBI injury, then treated with BHB after TBI injury. The sham injury mice still walked without falling. The TBI mice on day 1 showed the most foot steps, and by day 3, the number of foot steps in this group had dropped to about 50%. The pre-treatment group, the post-treatment group, the pre-and-post-treatment group, and the 2x pre-and-post-treatment group all showed improved beam walking on days 1 and 3. On day 3, all treatment groups improved significantly, with the best results observed in the Post-treatment and 2x Pre+Post-treatment groups, who showed a 70% improvement in footsteps one day after TBI.

[0111] [Figure 9A]The pg / μg total protein of IL-1β (FIG. 9A), IL-6 (FIG. 9B) and IL-10 (FIG. 9C) expressed in the cortex of each test group from FIG. 8 is shown. The results for TNF-α were not significant (data not shown). The data at 24 hours and 14 days in FIG. 9A, FIG. 9B and FIG. 9C show that treatment reduced the expression of IL-1β, IL-6 and IL-10. IL-10 is generally considered an anti-inflammatory cytokine, but can also promote fibrotic processes [Steen, EH et al. Adv. Wound Care (New Rochelle) (2020) 9(4): 184-98, citing O'Garra, A. et al. Immunol. Rev. (20008) 223: 114-31]. The data for the pre+post group are consistent with the behavioral data. [Figure 9B] The pg / μg total protein of IL-1β (FIG. 9A), IL-6 (FIG. 9B) and IL-10 (FIG. 9C) expressed in the cortex of each test group from FIG. 8 is shown. The results for TNF-α were not significant (data not shown). The data at 24 hours and 14 days in FIG. 9A, FIG. 9B and FIG. 9C show that treatment reduced the expression of IL-1β, IL-6 and IL-10. IL-10 is generally considered an anti-inflammatory cytokine, but can also promote fibrotic processes [Steen, EH et al. Adv. Wound Care (New Rochelle) (2020) 9(4): 184-98, citing O'Garra, A. et al. Immunol. Rev. (20008) 223: 114-31]. The data for the pre+post group are consistent with the behavioral data. [Figure 9C]The pg / μg total protein of IL-1β (FIG. 9A), IL-6 (FIG. 9B) and IL-10 (FIG. 9C) expressed in the cortex of each test group from FIG. 8 is shown. The results for TNF-α were not significant (data not shown). The data at 24 hours and 14 days in FIG. 9A, FIG. 9B and FIG. 9C show that treatment reduced the expression of IL-1β, IL-6 and IL-10. IL-10 is generally considered an anti-inflammatory cytokine, but can also promote fibrotic processes [Steen, EH et al. Adv. Wound Care (New Rochelle) (2020) 9(4): 184-98, citing O'Garra, A. et al. Immunol. Rev. (20008) 223: 114-31]. The data for the pre+post group are consistent with the behavioral data. Summary of the Invention

[0112] According to one aspect, the present disclosure provides a composition comprising an active ingredient encapsulated in a suspension of particles comprising a biodegradable polymer containing an effective amount of an active agent, the particles configured to bypass the blood-brain barrier, the active ingredient is synthetic D-beta hydroxybutyrate, and the targeted delivery is to the brain.

[0113] According to some embodiments, the size of the particles is greater than 10 nm in diameter, greater than 15 nm in diameter, greater than 20 nm in diameter, greater than 30 nm in diameter, greater than 40 nm in diameter, greater than 50 nm in diameter, greater than 60 nm in diameter, greater than 70 nm in diameter, greater than 80 nm in diameter, greater than 90 nm in diameter, greater than 100 nm in diameter, greater than 200 nm in diameter, greater than 300 nm in diameter, greater than 400 nm in diameter, greater than 500 nm in diameter. The diameter of the synthetic D-beta hydroxybutyrate is preferably greater than 600 nm, greater than 700 nm, greater than 800 nm, greater than 900 nm, greater than 1000 nm, greater than 2000 nm, greater than 3000 nm, greater than 4000 nm, greater than 5000 nm, greater than 6000 nm, greater than 7000 nm, greater than 8000 nm, greater than 9000 nm, or greater than 10,000 nm. According to some embodiments of the composition, the synthetic D-beta hydroxybutyrate is dispersed throughout the particle, or the particle is impregnated with the synthetic D-beta hydroxybutyrate, or the particle comprises a matrix, and the matrix comprises the synthetic D-beta hydroxybutyrate. According to some embodiments of the composition, the incorporation of the synthetic D-beta hydroxybutyrate into the polymer carrier can achieve controlled release, including delayed release and sustained release. According to some embodiments of the composition, the polymer comprises a mucoadhesive polymer, a thermoresponsive polymer, or both. According to some embodiments of the composition, a portion of the synthetic D-beta hydroxybutyrate is adsorbed or weakly bound to the surface of the particle, contributing to a rapid initial release or a burst release. According to some embodiments of the composition, the particle has any order of release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, extended release, or a combination thereof. According to some embodiments of the composition, the release of the synthetic D-beta hydroxybutyrate from the particle is via diffusion, erosion, or both. According to some embodiments of the composition, the composition is formulated for intranasal delivery.

[0114] According to some embodiments of the composition, the mucoadhesive polymer comprises chitosan, alginate and cellulose or derivatives thereof.

[0115] According to some embodiments of the composition, the thermoreversible polymer comprises gelatin, carrageenan, methylcellulose, hydroxypropylmethylcellulose (HPMC), xyloglucan, poly(N-isopropylacrylamide-c-acrylic acid), poly(N-isopropylacrylamide (PNIPAAm) / polyethylene oxide (PEO), poloxamer (Pluronic), or PEO / polylactic-co-glycolic acid (PLGA).

[0116] According to some embodiments of the composition, the polymer is selected from cellulose derivatives, polyacrylates, starches, gelatins, phospholipids, chitosan, poly-N-alkylacrylamides / poly-N-isopropylacrylamides, cyclodextrins, poloxamers, and methylcellulose.

[0117] According to some embodiments of the composition, when formulated for intranasal delivery, a maximum amount of about 25 mg / dose is administered to achieve a minimum daily dose ranging from about 85 mg / day to about 800 mg / day, inclusive.

[0118] According to another aspect, the disclosure provides a method for promoting brain health, comprising: (a) formulating a composition comprising an active ingredient as a suspension of particles comprising a biodegradable polymer containing an effective amount of the active ingredient; (b) administering to a subject the composition comprising the active ingredient, wherein the active ingredient is D-beta hydroxybutyrate; and (c) targeting the composition to the brain, wherein burst and controlled release of D-beta hydroxybutyrate can lead to effective D-beta hydroxybutyrate levels in the brain.

[0119] According to some embodiments of the method, the size of the particles is greater than 10 nm in diameter, greater than 15 nm in diameter, greater than 20 nm in diameter, greater than 30 nm in diameter, greater than 40 nm in diameter, greater than 50 nm in diameter, greater than 60 nm in diameter, greater than 70 nm in diameter, greater than 80 nm in diameter, greater than 90 nm in diameter, greater than 100 nm in diameter, greater than 200 nm in diameter, greater than 300 nm in diameter, greater than 400 nm in diameter, greater than 500 nm in diameter, greater than 500 nm in diameter, greater than 600 nm in diameter, greater than 700 nm in diameter, greater than 800 nm in diameter, greater than 90 ...600 nm in diameter, greater than 700 nm in diameter, greater than 800 nm in diameter, greater than 900 nm in diameter, greater than 100 nm in diameter, greater than 200 nm nm, greater than 600 nm in diameter, greater than 700 nm in diameter, greater than 800 nm in diameter, greater than 900 nm in diameter, greater than 1000 nm in diameter, greater than 2000 nm in diameter, greater than 3000 nm in diameter, greater than 4000 nm in diameter, greater than 5000 nm in diameter, greater than 6000 nm in diameter, greater than 7000 nm in diameter, greater than 8000 nm in diameter, greater than 9000 nm in diameter, or greater than 10,000 nm in diameter. According to some embodiments of the method, the synthetic D-beta hydroxybutyrate is dispersed throughout the particle, or the particle is impregnated with D-beta hydroxybutyrate, or the particle comprises a matrix, and the matrix comprises D-beta hydroxybutyrate. According to some embodiments of the method, the incorporation of the synthetic D-beta hydroxybutyrate into the polymer carrier can achieve controlled release, including delayed release and sustained release. According to some embodiments of the method, the polymer comprises a mucoadhesive polymer, a thermoresponsive polymer, or both. According to some embodiments of the method, a portion of the synthetic D-beta hydroxybutyrate is adsorbed or weakly bound to the surface of the particle, contributing to a rapid initial release or a burst release. According to some embodiments of the method, the particle has any order of release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, extended release, or a combination thereof. According to some embodiments of the method, the release of the synthetic D-beta hydroxybutyrate from the particle is via diffusion, erosion, or both. According to some embodiments of the method, the composition is formulated for oral or intranasal delivery.

[0120] According to some embodiments of the method, the mucoadhesive polymer comprises chitosan, alginate and cellulose or derivatives thereof.

[0121] According to some embodiments of the method, the thermoreversible polymer comprises gelatin, carrageenan, methylcellulose, hydroxypropylmethylcellulose (HPMC), xyloglucan, poly(N-isopropylacrylamide-c-acrylic acid), poly(N-isopropylacrylamide (PNIPAAm) / polyethylene oxide (PEO), poloxamer (Pluronic), or PEO / polylactic-co-glycolic acid (PLGA).

[0122] According to some embodiments of the method, the polymer is selected from cellulose derivatives, polyacrylates, starches, gelatins, phospholipids, chitosan, poly-N-alkylacrylamides / poly-N-isopropylacrylamides, cyclodextrins, poloxamers, and methylcellulose.

[0123] According to some embodiments of the method, when formulated for intranasal delivery, a maximum amount of about 25 mg / dose is administered to achieve a minimum daily dose ranging from about 85 mg / day to about 800 mg / day, inclusive.

[0124] According to another aspect, the disclosure provides a method for reducing the risk of brain damage due to acquired brain injury for a subject susceptible to or otherwise at risk for acquired brain injury, comprising administering a pharmaceutical composition comprising a pharma- ceutical carrier and a formulation comprising a therapeutic amount of an API, wherein the API is synthetic D-beta hydroxybutyrate, wherein the administering is intranasal (IN), and wherein the administering comprises targeted delivery to the olfactory region of the subject's nasal cavity, wherein the targeted delivery achieves an effective amount of the API in the brain, and wherein the effective amount of the pharmaceutical composition (i) eliminates or reduces the risk of acquired brain injury, or (ii) reduces the severity of acquired brain injury, or (iii) delays the onset of acquired brain injury, or (iv) a combination thereof.

[0125] According to some embodiments of the method, the acquired brain injury is traumatic brain injury. According to some embodiments of the method, the administering is neuroprotective. According to some embodiments of the method, the subject's risk of acquired brain injury is increased by participating in an event where acquired brain injury is a known risk. According to some embodiments of the method, the administering is within 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 24 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 hours, 74 hours, 75 hours, 76 hours, 77 hours, 78 hours, 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 83 hours, 84 hours, 85 hours, 86 hours, 87 hours, 8 and up to 25 mg / dose, such as within 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, etc. According to some embodiments of the method, the pharmaceutical composition comprises a liquid spray formulation or a dry powder formulation. According to some embodiments of the method, the pharmaceutical composition comprises an active agent formulated as a solution, suspension, or dispersion. According to some embodiments of the method, the minimum daily dose of the pharmaceutical composition contains about 85 mg / day to about 800 mg / day of API, inclusive.

[0126] According to some embodiments of the method, the traumatic brain injury comprises a concussion. According to some embodiments of the method, the event is a military operation or a sporting event. According to some embodiments of the method, the sporting event is hockey, football, soccer, baseball, polo, rugby, horse riding, car racing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing.

[0127] According to some embodiments of the method, the pharmaceutical composition is delivered as a liquid spray comprising a droplet size distribution comprising droplets containing the API. According to some embodiments of the method, the dry powder formulation when aerosolized comprises a cloud of very fine particles comprising the API. According to some embodiments of the method, the formulation is formulated with an excipient.

[0128] According to some embodiments of the method, the droplet size distribution of the liquid spray is in the range of about 20 μm to about 120 μm, inclusive, containing a therapeutic amount of the API. According to some embodiments of the method, the particle distribution is depleted of smaller particles that would otherwise enter the lungs.

[0129] According to another aspect, the disclosure provides a method for treating symptoms of brain injury including neuroinflammation in a subject comprising administering a pharmaceutical composition comprising a pharma- ceutically acceptable carrier and a formulation comprising a therapeutic amount of an API, wherein the API is synthetic D-beta hydroxybutyrate, and wherein the administering is intranasal (IN), and wherein the administering comprises targeted delivery to the olfactory region of the subject's nasal cavity, wherein the targeted delivery achieves an effective amount of the API in the brain, and wherein the effective amount of the pharmaceutical composition achieves one or more therapeutic effects including (i) reducing the severity of the acquired brain injury, or (ii) limiting the occurrence of symptoms characteristic of the acquired brain injury being treated, or (iii) limiting the worsening of symptoms characteristic of the acquired brain injury being treated, or (iv) limiting the recurrence of acquired brain injury in a subject who previously had acquired brain injury, or limiting the recurrence of symptoms in a subject who was previously asymptomatic for acquired brain injury, or a combination thereof.

[0130] According to some embodiments of the method, the acquired brain injury is a traumatic brain injury. According to some embodiments of the method, the administering is neuroprotective. According to some embodiments of the method, the risk of acquired brain injury in a subject is increased by participating in an event where acquired brain injury is a known risk. According to some embodiments of the method, administering is within 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 ... and 25 mg / dose for at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, at least 24 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., following the event of sustaining the acquired brain injury. According to some embodiments of the method, the pharmaceutical composition comprises a liquid spray formulation or a dry powder formulation. According to some embodiments of the method, the pharmaceutical composition comprises the active agent formulated as a solution, suspension or dispersion.According to some embodiments of the method, the minimum daily dose of the pharmaceutical composition contains about 85 mg / day to about 800 mg / day of the API, inclusive. According to some embodiments of the method, the traumatic brain injury comprises a concussion. According to some embodiments of the method, the event is a military operation or a sporting event. According to some embodiments of the method, the sporting event is hockey, football, soccer, baseball, polo, rugby, horse riding, car racing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing. According to some embodiments of the method, the pharmaceutical composition is delivered as a liquid spray comprising a droplet size distribution comprising droplets containing the API. According to some embodiments of the method, the dry powder formulation when aerosolized comprises a cloud of very fine particles containing the API. According to some embodiments of the method, the formulation is formulated with excipients. According to some embodiments of the method, the droplet size distribution of the liquid spray is in the range of about 20 μm to about 120 μm, inclusive, containing a therapeutic amount of the API.

[0131] According to another aspect, the disclosure provides a method for reducing the risk of acquired brain injury and treating symptoms of acquired brain injury after a subject suffers brain injury in a subject susceptible to or otherwise at risk for acquired brain injury, wherein the subject's risk of acquired brain injury is increased by participation in an event for which acquired brain injury is a known risk, comprising: (a) administering a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a formulation comprising a therapeutic amount of an API, wherein the API is synthetic D-beta hydroxybutyrate prior to the event, wherein administering is intranasal (IN) and neuroprotective, wherein administering comprises targeted delivery to the olfactory region of the subject's nasal cavity, wherein the targeted delivery achieves an effective amount of the API in the brain, and wherein the effective amount of the pharmaceutical composition (i) eliminates or reduces the risk of acquired brain injury. or (ii) reducing the severity of acquired brain injury, or (iii) delaying the onset of acquired brain injury, or (iv) a combination thereof; and (b) continuing intranasal (IN) administration after the event of brain injury, wherein an effective amount of the pharmaceutical composition achieves or continues to achieve one or more therapeutic benefits including (i') reducing the severity of the acquired brain injury, (ii') limiting the occurrence of symptoms characteristic of the acquired brain injury being treated, (iii') limiting the worsening of symptoms characteristic of the acquired brain injury being treated, (iv') limiting the recurrence of acquired brain injury in a subject who previously had acquired brain injury, or (v') limiting the recurrence of symptoms in a subject who was previously asymptomatic for acquired brain injury, (vi') or a combination thereof.

[0132] According to some embodiments of the method, the acquired brain injury is a traumatic brain injury. According to some embodiments of the method, the pharmaceutical composition comprises a liquid spray formulation or a dry powder formulation. According to some embodiments of the method, the pharmaceutical composition comprises an API formulated as a solution, suspension or dispersion. According to some embodiments of the method, the minimum daily dose of the pharmaceutical composition contains about 85 mg / day to about 800 mg / day of the API, inclusive. According to some embodiments of the method, the traumatic brain injury comprises a concussion. According to some embodiments of the method, the event is a military operation or a sporting event. According to some embodiments of the method, the sporting event is hockey, football, soccer, baseball, polo, rugby, horse riding, car racing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing.According to some embodiments of the method, administration may be as frequently as tolerated, for example, every 2 minutes, 5 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes following the event, for at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, for at least 1 day, for at least 2 days, for at least 3 days, for at least 4 days, for at least 5 days, for at least 6 days, for at least 7 days, for at least 8 days, for at least 9 days, for at least 10 days, for at least 11 days, for at least 12 days, for at least 13 days, for at least 14 days, for at least 15 days, for at least 16 days, for at least 17 days, for at least 18 days, for at least 19 days, for at least 20 days, for at least 21 days, for up to 25 days. mg / dose and as frequently as tolerated, for example, every 2, 5, 10, 20, 30, 40, 50, or 60 minutes after the brain injury for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or at least 24 hours, for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, or at least 21 days up to a maximum of 25 mg / dose.

[0133] According to some embodiments of the method, the pharmaceutical composition is delivered as a liquid spray comprising a droplet size distribution comprising droplets containing the API. According to some embodiments of the method, the dry powder formulation when aerosolized comprises a cloud comprising a distribution of fine particles comprising the API. According to some embodiments of the method, the formulation is formulated with an excipient. According to some embodiments of the method, the droplet size distribution of the liquid spray is in the range of about 20 μm to about 120 μm, inclusive, containing a therapeutic amount of the API. According to some embodiments of the method, the particle distribution is depleted of smaller particles that would otherwise enter the lungs.

[0134] According to another aspect, the disclosure provides a pharmaceutical composition for use in reducing the risk of brain injury due to acquired brain injury in a subject susceptible to or otherwise at risk for acquired brain injury, the composition comprising a pharma- ceutically acceptable carrier and a formulation comprising a therapeutic amount of an API, where the API is synthetic D-beta hydroxybutyrate, and the treatment comprises targeted delivery of 25 mg / dose to the olfactory region of the nasal cavity of the subject (a) before the subject participates in an event where acquired brain injury is a known risk, (b) after the subject participates in the event, or both, for use in treating a symptom of acquired brain injury in a subject including neuroinflammation.

[0135] According to some embodiments of the pharmaceutical composition for use, the targeted delivery achieves an effective amount of the API in the subject's brain. According to some embodiments, the effective amount of the pharmaceutical composition limits the occurrence of symptoms characteristic of acquired brain injury, and / or limits the worsening of symptoms characteristic of acquired brain injury, and / or limits the recurrence of acquired brain injury in a subject who previously had acquired brain injury, and / or limits the recurrence of symptoms in a subject who was previously asymptomatic for acquired brain injury.

[0136] According to some embodiments of the pharmaceutical composition for use, the acquired brain injury is a traumatic brain injury, and / or the brain injury comprises neuroinflammation, and / or the event is a sporting event, and / or the event is a military operation, and / or the administering is neuroprotective, and / or the pharmaceutical composition comprises an API formulated as a solution, suspension or dispersion, and / or the administering is a minimum amount of about 85 mg / day to about 800 mg / day of API, inclusive. According to some embodiments of the pharmaceutical composition for use, the traumatic brain injury comprises a concussion. According to some embodiments of the pharmaceutical composition for use, the sporting event is hockey, football, soccer, baseball, polo, rugby, horse riding, auto racing, motorcycling, skiing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0137] Glossary The term "aerodynamic diameter" as used herein refers to the diameter of a sphere of unit density that behaves aerodynamically in the same way as particles of a test substance / compound. It is used to predict where particles of different sizes and densities may be deposited in the airways.

[0138] As used herein, the term "active ingredient" refers to a drug, molecule, composition, or other substance that provides a biological activity or effect.

[0139] The term "active pharmaceutical ingredient" ("API" or "bulk active") is the substance in a drug that is pharma- ceutical active.

[0140] The term "adenosine 5'-triphosphate" or "ATP" is the primary molecule for storing and transporting energy within cells.

[0141] The term "administer" as used herein means to give or apply. The term "administer" as used herein includes in vivo administration and direct administration to tissue ex vivo. In general, the compositions may be administered systemically, either orally, bucally, parenterally, by inhalation or injection (i.e., through the mouth or nose), or rectally in dosage unit formulations containing conventional non-toxic pharma- ceutically acceptable carriers, adjuvants, and vehicles, if desired, or may be administered locally by means such as, but not limited to, injection, implantation, implantation, or topical application. Administration may be performed once, multiple times, and / or over one or more extended periods, for example, as individual unit doses or in the form of a treatment regimen that includes multiple unit doses of multiple drugs and / or substances.

[0142] As used herein, the term "adsorb" refers to adhesion to a surface.

[0143] The term "aerosol" as used herein refers to a substance that can be enclosed under pressure and expelled as a fine spray, typically by a propellant gas. For example, a colloidal dispersion of solid or liquid particles in a gas is an aerosol.

[0144] The term "aerosol generator" refers to a device that produces an aerosol.

[0145] As used herein, the term "aerosolized" refers to being dispersed or expelled in the form of a colloidal dispersion of solid or liquid particles in a gas.

[0146] Anatomical terms:

[0147] When referring to animals, they typically have one end with the head and mouth, and the opposite end often has the anus and tail, with the cephalic end referred to as the cranial end, while the caudal end is referred to as the caudal end. Within the head itself, rostral is used to refer to the direction toward the end of the nose, and caudal is used to refer toward the tail. The surface or side of an animal's body that normally faces upwards and away from the pull of gravity is the dorsal, and its opposite is typically the side closest to the ground when walking, swimming, or flying with all legs, the ventral. On a limb or other appendage, the point closest to the main body is the "proximal" and the point away is the "distal". Three basic reference planes are used in zoological anatomy. The "sagittal" plane divides the body into left and right portions. The "midsagittal" plane is at the midline, i.e., passes through midline structures such as the spine, and all other sagittal planes are parallel to it. The "coronal" plane divides the body into dorsal and ventral portions. The "transverse" plane divides the body into cranial and caudal portions.

[0148] When referring to humans, the body and its parts are always described using the assumption that the body is upright. The part of the body closer to the head end is the "superior" (corresponding to the cranium in animals), while the part away from it is the "inferior" (corresponding to the caudal side in animals). Objects closer to the front of the body are referred to as the "anterior" (corresponding to the ventral side in animals) and objects closer to the rear of the body are referred to as the "posterior" (corresponding to the dorsal side in animals). The transverse, axial, or horizontal plane is an XY plane parallel to the ground, separating the superior / head from the inferior / feet. The coronal or anterior plane is a YZ plane perpendicular to the ground, separating the anterior from the posterior. The sagittal plane is an XZ plane perpendicular to the ground and to the coronal plane, separating the left from the right. The midsagittal plane is a specific sagittal plane that is exactly in the middle of the body.

[0149] Structures near the midline are called medial, and structures near the sides of an animal are called lateral. Thus, medial structures are closer to the midsagittal plane and lateral structures are further from the midsagittal plane. Structures at the midline of the body are midline. For example, the tip of the nose of a human subject is at the midline.

[0150] Ipsilateral means on the same side, contralateral means on the other side, and bilateral means on both sides. Structures closer to the center of the body are proximal or central, while structures further away are distal or peripheral. For example, the hand is at the distal end of the arm, while the shoulder is at the proximal end.

[0151] The term "axon" as used herein refers to a nerve fiber that carries nerve impulses from a nerve cell to a target cell and also carries material from the nerve endings to the nerve cell.

[0152] The term "axonal injury" refers to the impairment and gradual loss of axons, which allow nerve cells to communicate with each other.

[0153] The terms "beta-hydroxybutyrate, β-hydroxybutyrate" and "BHB" are used interchangeably herein to refer to the predominant ketone body in the blood or synthetic BHB compound. In the body, it is formed primarily in the liver from free fatty acids during fasting or on a ketogenic diet, a high-fat, low-carbohydrate diet.

[0154] The term "binding" and other grammatical forms refer to a permanent attraction between chemical entities. Binding specificity involves both binding to a specific partner and not binding to other molecules. "Relative binding specificity" is a characteristic in biochemical systems where a molecule interacts differentially with its targets or partners, thereby affecting them distinctly depending on the identity of each individual target or partner.

[0155] The term "bioavailability" refers to the degree and rate at which an active moiety is absorbed from a drug product and becomes available at the site of action. 21 CFR 320.1. For products that enter the systemic circulation and thereby access the site of action, the surface charge on the polymeric particles influences the distribution of the particles throughout the body and the level of cellular uptake. [Vhora, I. et al. Applications of Polymers in Drug Delivery, Ch. 8, pages 221-61, at 234; Elsevier, Inc. (2021)]. Because cell membranes are negatively charged, positive charges favor higher intracellular concentrations. The exterior of the particles can be modified to bind targeting moieties to limit distribution at the target site. For example, PEG, PVP, and dextran are surface modifiers that prevent particles from being captured by the reticuloendothelial system (RES), thereby increasing their blood residence time (increasing their half-life) and improving their bioavailability. [Ibid.].

[0156] The term "biocompatible" as used herein refers to a material that is generally non-toxic to a recipient and has no significant adverse effects on the subject, and further, any metabolic or decomposition products of the material are non-toxic to the subject. Typically, a material that is "biocompatible" does not cause clinically relevant tissue irritation, damage, toxic reactions, or immunological responses against living tissue.

[0157] The term "biodegradable" as used herein refers to a material that erodes into soluble species or breaks down under physiological conditions into smaller units or chemical species that are themselves non-toxic (biocompatible) to the subject and can be metabolized, eliminated, or excreted by the subject. The two main types for biodegradation of delivered biodegradable polymeric materials are chemical degradation, e.g., by hydrolysis and oxidation-dependent degradation, and enzymatic degradation. Polymers can also be degraded by mechanical or thermal processes. [Vhora, I. et al. Applications of Polymers in Drug Delivery, Ch. 8, pages 221-61, at 228; Elsevier, Inc. (2021).

[0158] The term "bioequivalence" refers to the absence, in properly designed studies, of significant differences in the rate and extent to which the active ingredient or active moiety in a pharmaceutical equivalent or substitute becomes available at the site of drug action when administered in the same molar dose under similar conditions. 21 CFR 320.21.

[0159] The terms "biofermentation" or "microbial fermentation" are used interchangeably herein and refer to anaerobic biological processes in which the oxidation of a substrate is coupled to the reduction of another substrate or an intermediate derived from the oxidation. Fermenting bacteria are anaerobic but use organic molecules as the final electron acceptors to produce fermentation end products.

[0160] As used herein, the term "blood-brain barrier" ("BBB") refers to the network of blood vessels and tissues that are composed of tightly spaced cells and help prevent harmful substances in the blood from reaching the brain.

[0161] As used herein, the term "brain injury" refers to injury that causes destruction or deterioration of brain cells. According to some embodiments, brain injury includes neuroinflammation.

[0162] The term "brain health", as defined by the World Health Organization, refers to the state of the brain functioning across cognitive, sensory, social-emotional, behavioral and motor domains that enable people, with or without disabilities, to reach their full potential throughout their lifespan.

[0163] The term "carrier" as used herein describes a material that does not cause significant irritation to an organism and does not abolish the biological activity and properties of the compounds of the composition of the present invention described. The carrier must be of sufficiently high purity and sufficiently low toxicity to be suitable for administration to the mammal being treated. The carrier may be inert or may have pharmaceutical benefits. The terms "excipient", "carrier", or "vehicle" are used interchangeably to refer to carrier materials suitable for formulation and administration of the pharma-ceutically acceptable compositions described herein. Carriers and vehicles useful herein include any such materials known in the art that are non-toxic and do not interact with other components.

[0164] The term "carryover" or "CO" as used herein refers to a type of statistical error in HPLC-based analytical methods that originates from a previous sample and is introduced into a subsequent sample.

[0165] As used herein, the term "central nervous system" ("CNS") refers to the portion of the nervous system that consists of the brain and spinal cord.

[0166] The term "cerebrospinal fluid" as used herein refers to the ultrafiltrate of blood plasma contained within the ventricles of the brain and the subarachnoid spaces of the skull and spine. It is secreted primarily by the choroid plexus. A constant volume of secretion contributes to completing CSF renewal 4-5 times per 24-hour period in the average young adult.

[0167] As used herein, the term "chemical bond" refers to the attractive forces between atoms that are strong enough to allow the combined aggregate to function as a unit.

[0168] The term "chiral" is used to describe asymmetric molecules that are non-superimposable because they are mirror images of one another and therefore possess the property of chirality. Such molecules, also called enantiomers, are characterized by optical activity.

[0169] The term "chirality" refers to the geometric property of a rigid object (or a spatial arrangement of points or atoms) that it cannot be superimposed on its mirror image. If the object can be superimposed on its mirror image, the object is described as achiral.

[0170] The term "axis of chirality" refers to the axis along which a set of ligands are held to result in a spatial configuration that cannot be superimposed on its mirror image.

[0171] The term "chirality center" refers to an atom that holds a set of ligands in a spatial configuration that cannot be superimposed on its mirror image. A chirality center may be considered a generalized extension of the concept of an asymmetric carbon atom to the central atom of any element.

[0172] As used herein, the term "closed head injury" refers to an impact to the head from an external force that does not involve fracture or displacement of the skull. The term "open head injury" refers to trauma to the brain resulting from entry of a foreign object, such as a bullet, into the brain, resulting in loss of consciousness.

[0173] As used herein, the term "compatible" means that the components of the composition are capable of being combined with each other in a manner such that there are no interactions that would substantially reduce the effectiveness of the composition under normal use conditions.

[0174] As used herein, the term "composition" refers to a mixture of components.

[0175] As used herein, the term "concussion" refers to the disruption of neurological function resulting from a head injury or violent shaking.

[0176] The term "condition" as used herein refers to a variety of health conditions and is meant to include any underlying mechanism or disorder, injury-induced disorder or disease, as well as the promotion of healthy tissues and organs.

[0177] The term "configuration" as used herein refers to the stereochemical arrangement of atoms that cannot be changed without breaking chemical bonds.

[0178] The term "conformation" refers to the geometric arrangement of atoms within a polymer chain.

[0179] As used herein, the term "contacting" and its various grammatical forms refer to the state or condition of touching, or of being in close proximity or local proximity, and includes contacting a composition with a target object.

[0180] The term "container closure system" as used herein refers to the collection of packaging components that together contain, protect and deliver the dosage form, including pumps for nasal and inhalation sprays.

[0181] The term "controlled release" is intended to refer to any active agent-containing formulation in which the manner and profile of active agent release from the formulation is controlled. This includes immediate release and non-immediate release formulations, which include, but are not limited to, sustained release and delayed release formulations. The term "delayed release" is used herein in its conventional sense to refer to an active agent formulation in which there is a time delay between administration of the formulation and the release of the active agent from the formulation. "Delayed release" may or may not involve gradual release of the active agent over an extended period of time, and thus may or may not be a "sustained release". As used herein, the term "extended" release means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 7 days, preferably about 30 to about 60 days. The term "sustained release" (also referred to as "sustained release") is used herein in its conventional sense to refer to an active agent formulation that provides for gradual release of the active agent over an extended period of time, preferably, but not necessarily, resulting in substantially constant blood levels of the active agent over an extended period of time.

[0182] The term "cytokine" as used herein refers to small soluble protein substances secreted by cells that have a variety of effects on other cells. Cytokines mediate many important physiological functions, including growth, development, wound healing, and immune response. They act by binding to cell-specific receptors located on the cell membrane, which allows distinct signaling cascades to be initiated within the cell, ultimately leading to biochemical and phenotypic changes in the target cell. Cytokines can act both locally and remotely from the site of release. Cytokines include type I cytokines, which encompass many of the interleukins as well as several hematopoietic growth factors, type II cytokines including interferons and interleukin-10, tumor necrosis factor ("TNF")-related molecules including TNFα and lymphotoxin, immunoglobulin superfamily members including interleukin-1 ("IL-1"), and chemokines, a family of molecules that play important roles in a wide variety of immune and inflammatory functions. The same cytokine may have different effects on cells depending on the state of the cell. Cytokines often regulate the expression of other cytokines, triggering cascades. Non-limiting examples of cytokines include, for example, IL-1 alpha, IL-beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 / IL-23 P40, IL13, IL-15, IL-17, IL-18, IL-21, IL-23, TGF-beta, IFN-gamma, GM-CSF, Gro. alpha, MCP-1, and TNF-alpha.

[0183] The term "D-value" or "mass division diameter" as used herein refers to the diameter that divides the mass of a sample into a specified percentage when all particles in the sample are arranged in ascending order of mass. The percentage of mass less than the diameter of interest is the number represented after "D". For example, a D10 diameter is the diameter where 10% of the mass of the sample is made up of smaller particles, and a D50 is the diameter where 50% of the mass of the sample is made up of smaller particles. D50 is also known as the "mass median diameter" because it divides the sample equally by mass. A D90 diameter is the diameter where 90% of the mass of the sample is made up of smaller particles. Although the D-value is based on dividing the mass of the sample by the diameter, the actual mass of the particles or sample does not need to be known. Since the D-value relates only to the ratio of masses, the relative mass is sufficient. This allows the use of optical measurement systems without the need to weigh the sample.

[0184] From the diameter values ​​obtained for each particle, the relative masses can be assigned according to the following relationship:

[0185] Mass of sphere = π / 6 d3 ρ

[0186] Assuming that ρ is a constant for all particles, and cancelling all constants from the equation, we get

[0187] Relative mass = d3

[0188] That is, the diameter of each particle is thus cubed to give its relative mass. These values ​​can be summed to calculate the total relative mass of the sample being measured. The values ​​may then be arranged in ascending order and added repeatedly until their sum reaches 10%, 50% or 90% of the total relative mass of the sample. The corresponding D value for each of these is the diameter of the last particle added to reach the required mass percentage.

[0189] As used herein, the term "delivery" refers to the carrying of an active agent to a location within the body.

[0190] As used herein, the term "derived from" is meant to encompass any method of receiving, obtaining, or modifying something from an original source.

[0191] "Diastereoisomers" are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.

[0192] As used herein, the term "diffusion" refers to the movement of individual molecules of a substance through a semipermeable barrier from a region of higher concentration to a region of lower concentration.

[0193] The term "disease" or "disorder" as used herein refers to an impaired health or state of abnormal function.

[0194] The term "distributed" as used herein refers to the act or process of distributing something over a wide area.

[0195] The term "dispersion" as used herein refers to a two-phase system, i.e., a system in which one phase is distributed as droplets within a second continuous phase. In these systems, the dispersed phase is often referred to as the discontinuous or internal phase, and the continuous phase is referred to as the external phase and comprises the continuous process medium. For example, in a coarse dispersion, the particle size is 0.5 μm. In a colloidal dispersion, the dispersed particles range in size from approximately 1 nm to 0.5 μm. A molecular dispersion is a dispersion in which the dispersed phase consists of individual molecules; if the molecules are smaller than colloidal size, the result is a true solution.

[0196] The term "dissolution" as used herein refers to the process by which a substance forms a solution. The "rate of dissolution" is the amount of active substance that goes into solution per unit time under standardized conditions of liquid / solid interface, temperature and solvent composition.

[0197] As used herein, the term "dose" refers to a predetermined amount of active agent administered at one time.

[0198] The term "dosage" as used herein refers to a predetermined amount and rate of administration of an active agent. Dosage level is based on a variety of factors, including the type of injury, the patient's age, weight, sex, medical condition, the severity of the condition, the route of administration, and the specific active agent used. Thus, dosing regimens can vary widely, but can be routinely determined by a physician using standard methods.

[0199] The term "dosage form" refers to the physical form that contains an active agent in combination with an excipient. Examples include solid dosage forms (tablets, capsules, pellets, pills, lozenges, granules, etc.), liquid dosage forms (solutions, suspensions, emulsions, elixirs, etc.), semi-solid dosage forms (ointments, creams, pastes, etc.), and gaseous dosage forms (aerosols, injections, etc.).

[0200] The term "dosing regimen" as used herein refers to the frequency with which active agent is administered, including formulation, route of administration, dosage, administration interval and administration period.The total daily dosage is calculated from the dosage and the number of times per day that dosage is administered.Designing the correct dosing regimen is important to achieve desired efficacy and avoid undesirable effects.

[0201] The term "droplet" as used herein refers to small droplets and means a very small volume of a flowable material. Factors that affect the size of the droplets are pressure, viscosity, fluid temperature, and surface tension. For example, increasing pressure, increasing fluid temperature, increasing viscosity, and increasing surface tension decrease the droplet size.

[0202] The term "droplet size distribution" as used herein refers to a statistical measure of the percentage of droplets of a particular size (or a particular size interval). A simple distribution function assumes that all droplets have the same shape (usually spherical), which is generally valid for a fully developed spray. Such a distribution is fully characterized by (1) a maximum diameter, (2) a size parameter, and (3) a distribution parameter, which can be calculated from any three known independent average diameters. The remaining problem consists of correlating these parameters with the physical characteristics of the spray generation system, including the physical properties of the droplet phase and the continuous medium. Among these properties are density, viscosity, and interfacial tension. [Mugele, RA and Evans, HD. Droplet size distribution in sprays. Industry. And Engineering Chemistry (1951) 43(6):1317-24].

[0203] The terms "drug loading (%)" and "drug loading capacity" are used interchangeably herein to refer to the ratio of the weight of a compound of the present invention in a microparticle to the total weight of the microparticle, expressed as a percentage, which reflects the drug content of the microparticle.

[0204] The term "drug product" as used herein refers to a finished dosage form that generally, though not necessarily, contains a drug substance in association with one or more other ingredients.

[0205] The term "drug substrate" as used herein refers to an active ingredient intended to provide pharmacological activity or other direct effect in the diagnosis, cure, mitigation, treatment, or prevention of disease, or to affect the structure and function of the body, but does not include intermediates used in the synthesis of such ingredients.

[0206] The term "effective amount" refers to the amount of the described pharmaceutical composition of the invention necessary or sufficient to realize a desired biological effect. In addition, the term "effective amount" includes preventative or prophylactic amounts of the described compositions of the invention. In preventative or prophylactic applications of the described invention, the pharmaceutical composition or medicament is administered to a patient susceptible to or otherwise at risk of a disease, disorder or condition in an amount sufficient to eliminate or reduce the risk, reduce the severity, or delay the onset of a disease, disorder or condition, including biochemical, histological, and / or behavioral symptoms of the disease, disorder or condition, complications thereof, and intermediate pathological phenotypes exhibited during the development of the disease, disorder or condition.

[0207] The term "emulsion" as used herein refers to a two-phase system prepared by combining two immiscible liquid carriers, one of which is distributed evenly throughout the other. Emulsions consist of globules with diameters equal to or greater than that of the largest colloidal particle. The phase present as small droplets is called the dispersed phase, and the surrounding liquid is known as the continuous phase. The size of the globules should be such that the system achieves maximum stability. Separation of the two phases usually occurs unless a third substance (emulsifier) ​​is incorporated.

[0208] "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon may be specified by either R or S. A resolved compound of unknown absolute configuration may be designated as (+) or (-), depending on the direction (dextrorotatory or levorotatory) that the compound rotates plane-polarized light at the wavelength of the sodium D line.

[0209] As used herein, the term "endogenous" refers to relating to or developed from factors within the body.

[0210] As used herein, the term "entrapment efficiency (%)" refers to the ratio of drug retained by the microparticles to the total amount available, expressed as a percentage.

[0211] As used herein, the term "erosion" refers to the loss of material from a polymer mass.

[0212] The term "excipient" is used herein to include any other agent or compound that may be included in the formulation that is not an active agent. Thus, the excipient must be pharma- ceutically or biologically acceptable or relevant (e.g., the excipient must generally be non-toxic to the subject). "Excipient" is intended to include a single such compound, as well as a plurality of such compounds.

[0213] The term "exogenous" as used herein refers to relating to or developing from factors outside the body.

[0214] The term "formulation" as used herein refers to a mixture prepared according to a particular procedure or a formula prepared according to a particular recipe or rules.

[0215] As used herein, the term "G protein" refers to specialized proteins that have the ability to bind the nucleotides guanosine triphosphate (GTP) and guanosine diphosphate (GDP).

[0216] The term "G protein-coupled receptor" (GPCR) refers to membrane receptors in eukaryotes that allow cells to receive information from their environment. GPCRs consist of a single polypeptide that folds into a globular shape and is embedded in the plasma membrane of the cell. Seven segments of the molecule span the entire width of the membrane (explaining why GPCRs are sometimes called seven-transmembrane receptors), and its intervening parts loop both inside and outside the cell. The extracellular loops form part of the pocket where signaling molecules bind to the GPCR. GPCRs interact with G proteins in the plasma membrane. When an external signaling molecule binds to the GPCR, it triggers a conformational change in the GPCR. This change then triggers an interaction between the GPCR and a nearby G protein. The G proteins that associate with GPCRs are heterotrimers, which means that they have three different subunits, namely, an alpha subunit, a beta subunit, and a gamma subunit. Two of these subunits (alpha and gamma) are attached to the plasma membrane by lipid anchors. G protein alpha subunits bind either GTP or GDP depending on whether the protein is active (GTP) or inactive (GDP). In the absence of a signal, GDP binds to the alpha subunit and the entire G protein-GDP complex binds to a nearby GPCR. This arrangement persists until a signaling molecule connects with the GPCR. At this point, a conformational change in the GPCR activates the G protein and GTP physically displaces the GDP bound to the alpha subunit. As a result, the G protein subunit dissociates into two parts: the GTP-bound alpha subunit and a beta-gamma dimer. Both parts remain anchored to the plasma membrane, but because they are no longer bound to the GPCR, they can diffuse laterally and interact with other membrane proteins. G proteins remain active as long as their alpha subunits connect with GTP. However, if this GTP is hydrolyzed back to GDP, the subunits again assume the form of an inactive heterotrimer and the entire G protein reassociates with the now inactive GPCR.In this way, G proteins function like switches that are turned on or off by signal-receptor interactions on the cell surface. Whenever a G protein is active, both its GTP-binding alpha subunit and its beta-gamma dimer can relay messages within the cell by interacting with other membrane proteins involved in signal transduction. Specific targets of activated G proteins include various enzymes that produce second messengers, as well as certain ion channels that allow ions to function as second messengers. Some G proteins stimulate the activity of these targets, while others are inhibitory. [Li, J. et al. The Molecule Pages database. Nature (2002) 420: 716-17].

[0217] The terms "glial cells", "neuro-glial cells" or "glia" refer to small cells lacking axons and / or dendrites present in the central and peripheral nervous systems. Glial cells act as regulators of the CNS and PNS environment, increasing and decreasing activity within synapses by regulating the uptake of neurotransmitters, oxygen, and ions, and aiding in recovery from neuronal injury. Macroglia (which include astrocytes and oligodendrocytes) insulate and protect neurons, and aid in their development and migration. Microglial cells are smaller cells derived from hematopoietic stem cells (although some may be derived directly from neural stem cells). They share many properties with tissue macrophages, and are primarily scavenger cells that remove cellular debris from sites of injury or normal cellular transformation. In fact, some neurobiologists prefer to classify microglia as a type of macrophage. After brain injury, the number of microglia at the site of injury increases dramatically. Some of these cells grow from microglia, which are resident in the brain, while others are derived from macrophages that migrate from the circulation to the area of ​​injury. Furthermore, microglia and macroglia cooperate to ensure optimal neuronal function and neurotransmission through synapses. [See Purves D,et al.,Ed.(2001).“Neuroglial Cells.”Neuroscience.2nd edition.(Retrieved from https: / / www.ncbi.nlm.nih.gov / books / NBK10869 / )

[0218] As used herein, the term "glial fibrillary acidic protein" or "GFAP" refers to a marker of astrocyte injury that is a marker for predicting TBI. Several mouse model studies have been conducted to assess functional outcomes after TBI. Increased GFAP levels correlate with impaired spatial learning, as evidenced by impaired performance in the maze test [Brennan, J. et al. Biomarkers for Traumatic Brain Injury, Chapter 26-Neuropsychological testing, Editors: Alan HB Wu, W. Peacock, Academic Press (2020), pp. 397-409, citing Ferguson, S. et al. J. Neurotrauma (2016) 34(8): 1676-91; Marschner, L. et al. Behav. Brain Res. (2019) 365: 222-30; Broussard, JI et al. Brain In. (2018) 32(1): 113-22]. Human clinical trials focusing on TBI outcome and GFAP have been conducted, and GFAP was found to be significantly associated with Glasgow Outcome Score (GOS) [Ibid. citing Nylen, K. et al. J. Neurol. Sci. (2006) 240(1):85-91].

[0219] As used herein, the term "HCA2" refers to the G protein-coupled receptor for BHB and nicotinic acid.

[0220] As used herein, the term "healthy control" refers to a subject who has no symptoms or other clinical evidence of TBI.

[0221] As used herein, the term "immune cells" refers to cells that are part of the immune system and help the body fight infections and other diseases. Immune cells include lymphocytes (e.g., B cells, T cells, natural killer (NK) cells), neutrophils, and monocytes / macrophages.

[0222] The term "immune system" as used herein refers to the body's defense system against disease, including the innate immune system and the adaptive immune system. The innate immune system provides a non-specific first line of defense against pathogens. It includes physical barriers (e.g., skin) and both cellular (granulocytes, natural killer cells) and humoral (complement system) defense mechanisms. The response of the innate immune system is immediate, but unlike the adaptive immune system, it does not provide lasting immunity against pathogens. The adaptive immune response is the response of the immune system of a vertebrate animal to a specific antigen that typically generates immune memory.

[0223] The term "impregnated" as used herein in its various grammatical forms refers to infusing or permeating throughout, thereby filling voids and / or saturating with a substance.

[0224] As used herein, the term "impurity profile" refers to a description of the identified and unidentified impurities present in an API.

[0225] The term "intermediate" as used herein refers to a material produced during a step in the processing of an API that undergoes further molecular changes or purification before becoming the API. Intermediates may or may not be isolated.

[0226] The term "inflammation" as used herein refers to the physiological process by which vascularized tissue responds to injury. See, for example, FUNDAMENTAL IMMUNOLOGY, 4th Ed., William E. Paul, ed. Lippincott-Raven Publishers, Philadelphia (1999) at 1051-1053, which is incorporated herein by reference. During the inflammatory process, cells involved in detoxification and repair are recruited to the damaged site by inflammatory mediators. Inflammation is often characterized by a strong infiltration of leukocytes, especially neutrophils (polymorphonuclear cells), at the site of inflammation. These cells promote tissue damage by releasing toxic substances into the blood vessel wall or uninjured tissue. Traditionally, inflammation is divided into acute and chronic responses.

[0227] As used herein, the term "acute inflammation" refers to a rapid, short-lived (minutes to days), relatively uniform response to acute injury characterized by the accumulation of fluid, plasma proteins, and neutrophilic leukocytes. Examples of harmful agents that cause acute inflammation include, but are not limited to, pathogens (e.g., bacteria, viruses, parasites), foreign bodies from exogenous (e.g., asbestos) or endogenous (e.g., uric acid crystals, immune complexes), sources, and physical (e.g., burns) or chemical (e.g., caustic) agents.

[0228] The term "chronic inflammation" as used herein refers to inflammation that is longer in duration and has a vague or indefinite termination. Chronic inflammation takes over when acute inflammation persists due to incomplete clearance of the initial inflammatory agent or as a result of multiple acute events occurring at the same location. Chronic inflammation, including the influx of lymphocytes and macrophages and the growth of fibroblasts, can cause tissue scarring at the site of prolonged or repeated inflammatory activity.

[0229] The term "injury," as used herein, refers to damage or harm to a structure or function of the body caused by an external agent or force, which may be physical or chemical.

[0230] As used herein, the term "infusion" refers to the act of delivering air, gas, or powder under pressure into a cavity or cavity of the body. For example, nasal infusion refers to the act of delivering air, gas, or powder under pressure through the nose.

[0231] The term "isomer" as used herein refers to one of two or more molecules that have the same number and type of atoms and therefore the same molecular weight, but differ in chemical structure. Isomers may differ in the connectivity of the atoms (structural isomers) or may have the same atomic connectivity but differ only in the arrangement or configuration of the atoms in space (stereoisomers). Stereoisomers include, but are not limited to, double bond isomers, enantiomers, and diastereomers. Enantiomers are mirror images that are not superimposable. A mixture of equal parts of the optical forms of a compound is known as a racemic mixture or racemate. Diastereomers are stereoisomers that are not mirror images. Stereoisomers may include enantiomers, diastereomers, or E or Z alkene, imine, or oxime isomers. Stereoisomeric mixtures include racemic mixtures, diastereomeric mixtures, or E / Z isomeric mixtures. Stereoisomers can be synthesized in pure form (Nogradi, M.; Stereoselective Synthesis, (1987) VCH Editor Ebel, H. and Asymmetric Synthesis, Volumes 3-5, (1983) Academic Press, Editor Morrison, J.) or resolved by various methods such as crystallization and chromatographic techniques (Jaques, J.; Collet, A.; Wilen, S.; Enantiomer, Racemates, and Resolutions, 1981, John Wiley and Sons and Asymmetric Synthesis, Vol. 2, 1983, Academic Press, Editor Morrison, J). Unless otherwise specified, the formulations of the present disclosure are meant to include all such possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents or resolved using conventional techniques. If the compound contains a double bond, the substituent may be in the E or Z configuration.If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have either the cis or trans configuration. All tautomeric forms are intended to be included.

[0232] As used herein, the term "interleukin" or "IL" is the generic name for a cytokine produced by white blood cells.

[0233] The term "interleukin-1β" ​​or "IL-1β" as used herein refers to a member of the IL-1 family of inflammatory cytokines. As of 2019, human sequence algorithm technology suggests that the IL-1 family contains a total of 11 members with similar or different biological effects [32, 33]. IL-1α, IL-1β, IL-1Ra, IL-18, IL-33, IL-36α, IL-36β, IL-36γ, IL-36Ra IL-37, and IL-38 have been identified and characterized [Kaneko, N. et al. Inflammation and Regeneration (2019) 39: article 12, citing Dinarello, C. et al. Nature Immunol. (2010) 11: 973]. Among them, IL-1α, IL-1β, IL-18, IL-33, and IL-36 are receptor agonistic, while IL-1Ra, IL-36Ra, and IL-38 are receptor antagonistic. IL-37 is the only anti-inflammatory cytokine. There are two individual forms of IL-1, IL-1α and IL-1β, isolated from two different cDNAs, which cannot be distinguished in terms of their biological functions [ibid., citing Lachman, LB et al. J. Immunol. 91977] 119:2019-23]. Although the homology between IL-1α and IL-1β is not high (27%) in terms of amino acid sequence, IL-1α and IL-1β are structurally similar and exhibit the same functions by sharing a common receptor, the type 1 receptor for IL-1 (IL-1R1). IL-1R1 initiates inflammatory responses upon binding to the ligands IL-1α and IL-1β, and is reported to be expressed by T lymphocytes, fibroblasts, epithelial cells, and endothelial cells.

[0234] The term "interleukin-6" or "IL-6" as used herein refers to a pleiotropic cytokine that is not only involved in immune responses, but also in inflammation, hematopoiesis, bone metabolism, embryonic development, and other fundamental processes [Hirano, T. Intl Rev. Immunol. (1998) 16:249; Hunter, CA and Jones, SA. Nat. Immunol. (2015) 16:448; Van Snick, J. Annu. Rev. Immunol. (1990) 8:253; Heinrich, PC et al. Biochem. J. (2003) 374(Pt. 1):1; Kamimura, D. et al. Rev. Physiol. Biochem. Pharmacol. (2003) 149:1; Hasegawa, H. et al. Front. Immunol. (2016) 7:479]. IL-6 is the prototype member of the IL-6 family of cytokines, which consists of 10 members including IL-6, IL-11, IL-27, oncostatin M (OSM), leukemia inhibitory factor (LIF), ciliary neurotrophic factor (CNTF), cardiotrophin 1 (CT-1), cardiotrophin-like cytokine factor 1 (CLCF1), IL-35 and IL-39 [ibid., citing Murakami, M. et al. Immunity (2019) 50:812]. IL-6 is involved in cellular senescence, is produced by senescent cells, and plays an important role in senescence-induced inflammation and age-dependent pathologies, as well as cancer [ibid., citing Kuilman, T. et al. Cell (2008) 133:1019].IL-6 is also an important factor in inflammation, autoimmunity and cancer, and its effects are mainly exerted through the IL-6 signal transducer and activator of transcription 3 (STAT3) pathway [Ibid., citing Grivennikov, S. et al. Cancer Cell (2009) 15:103; Yu;, H. et al. Nat. Rev. Cancer (2009) 9:798; Jones, SA and Jenkins, BJ. Nat. Rev. Immunol. (2018) 18:773; Hirano, T. et al. Oncogene (2000) 19:2548; Jenkins, BJ et al. Blood (2007) 109:2380].

[0235] The term "interleukin 10" or "IL-10" as used herein refers to a pleotropic cytokine produced by nearly all species of activated immune cells, including B cells, mast cells, granulocytes (e.g., neutrophils, basophils, eosinophils), macrophages, dendritic cells, and multiple T cell subsets. [Citing Steen, EH et al. Adv. Wound Care (New Rochelle) (2020) 9(4): 184-98, O'Garra, A. et al. Immunol. Rev. (20008) 223: 114-31]. Its primary actions are believed to be primarily anti-inflammatory, inhibitory, or autoregulatory, and IL-10 appears to be a potent negative feedback regulator that influences the control and resolution of inflammation by autocrine and paracrine mechanisms. Although this immunosuppressive effect is broad and occurs at both the cellular and humoral levels, there are two primary means by which IL-10 limits potentially harmful inflammatory responses: (1) inhibition of antigen presentation by dendritic cells, and (2) inhibition of macrophage activation and infiltration to sites of injury, with secondary effects of attenuating proinflammatory cytokine expression. [Ibid., citing O'Garra, A. et al. Immunol. Rev. (20008) 223:114-31]. At the cellular level, IL-10 is thought to act as a posttranscriptional regulator that suppresses the messenger RNA (mRNA) stabilizing protein HuR (human antigen R), promoting specific destabilization of inflammatory cytokine mRNAs. [Ibid., citing Willis-Karp, M. et al. Mucosal Immunol. (2010) 3:104-10]. In addition, IL-10 is thought to inhibit apoptotic signaling pathways, such as the p38 MAPK (mitogen-activated protein kinase) pathway, through signal transducer and activator of transcription 3 (STAT3)-dependent signaling, thereby limiting tissue death and organ dysfunction following injury.[Ibid., citing Rajasingh, J. et al. FASEB J. (2006) 20:2112-14; Krishnamurthy, P. et al. Circ. Res. (2009) 104:e9-e18; Krishnamurthy, P. et al. FASEB J. (2010) 24:2484-94]. IL-10 signals through a tetrameric receptor complex (IL-10R) composed of two identical binding subunits IL-10Rα and two homologous signaling IL-10Rβ subunits. Some studies hypothesize that IL-10 inhibits fibrosis primarily by regulating inflammatory processes thought to lead to fibroproliferation, although the molecular mechanisms behind this described effect remain to be fully characterized. For example, prolonged IL-10 exposure or application to chronic disease processes may actually exacerbate tissue damage and promote fibrotic outcomes, despite being primarily beneficial in acute inflammatory settings and in the early stages of wound healing.

[0236] As used herein, the term "ketogenic diet" refers to a diet high in fat and low in carbohydrates (sugars) that causes the body to break down fat into molecules called ketones, which circulate in the blood and serve as the primary source of energy for many cells in the body.

[0237] The term "label" as used herein refers to a traceable moiety that is incorporated into a molecule in order to spatially locate it or follow it through a reaction or purification scheme. As a verb, it means to add such a group or atom that can be detected or measured.

[0238] As used herein, the term "labile" refers to something that undergoes increased decomposition.

[0239] The terms "LLOQ" or "Lower Limit of Quantitation" are used interchangeably and refer to the lowest amount of analyte in a sample that can be quantitatively determined with suitable precision and accuracy. Bioanalytical assays are validated within a given range, so a result below the LLOQ is below the limit of quantitation (BLQ).

[0240] The term "logS" as used herein is the base 10 logarithm of the solubility measured in mol / l, a common unit for measuring solubility, i.e., logS=log(solubility measured in mol / l).

[0241] As used herein, the term "macrophage" refers to large mononuclear phagocytic cells present in most tissues that have many functions, including as scavenger cells, pathogen recognition cells, and producers of pro-inflammatory cytokines.

[0242] The term "microglia" as used herein refers to the smallest of the glial cells that can function as phagocytes, cleaning up CNS debris. They are considered a type of immune cell found in the brain. Microglia are close relatives of other phagocytes, including macrophages and dendritic cells. Like macrophages, microglia originate from myeloid progenitor cells from the bone marrow. During embryonic development, these cells migrate to the CNS where they differentiate into microglia.

[0243] The term "modulate" as used herein refers to adjusting, altering, adapting, or adjusting to a particular measure or proportion.

[0244] The term "mutation" refers to a change in the DNA sequence within an organism's genes or chromosomes, or the process by which such changes occur in chromosomes, either by a change in the nucleotide sequence of the DNA encoding the gene, or by a change in the physical arrangement of the chromosome, resulting in the production of a new characteristic or trait not found in the parental type. Three mechanisms of mutation include substitutions (exchange of one base pair for another), additions (insertion of one or more bases into a sequence), and deletions (loss of one or more base pairs).

[0245] The term "nasal spray" as used herein refers to a drug product containing active ingredients dissolved or suspended in a typically aqueous-based formulation, which may contain other excipients, intended for use by nasal inhalation. A container closure system for a nasal spray includes the container and all of the components responsible for metering, atomizing, and delivering the formulation to the subject.

[0246] The term "neurofilament light" or "NfL" as used herein refers to a recognized biomarker of subcortical large axon degeneration [Mielke, MM et al. Neurology (2019) 93(3):e252-260; Hoffman, PN et al. Proc. Natl Acad. Sci. USA (1987) 84:3472-6; Norgren, N. et al. Brain Res. (2003) 987:25-31]. CSF and plasma NfL levels are elevated in multiple neurodegenerative disorders including Alzheimer's disease (AD dementia) [ibid., citing Mattsson, N. et al. JAMA Neurol. (2017) 74:557-66; Kern, S. et al. JAMA Neurol. (2019) 76(2):187-933], frontotemporal dementia [ibid., citing Scherling, CS et al. Ann. Neurol. (2014) 75:116-26], multiple sclerosis [ibid., citing Teunissen, CE et al. Lancet Neurol. (2005) 4:32-41], and TBI.

[0247] As used herein, the term "neuroinflammation" refers to an inflammatory response in the central nervous system mounted by peripheral immune and glial cells, during which the permeability of the BBB is often increased.

[0248] As used herein, the term "neuroprotection" refers to helping protect nerve cells from damage, degeneration, or dysfunction.

[0249] The term "nicotinic acid" as used herein refers to B complex vitamins (such as nicotinamide) that prevent pellagra. The daily requirement is 15-20 mg for adults. At higher doses (0.5-2 g), nicotinic acid can activate the HCA2 receptor.

[0250] As used herein, the phrase "nutraceutically acceptable carrier" refers to any substantially non-toxic carrier that can be used to formulate and administer the compositions of the present invention described, in which the products of the present invention described are stable and bioavailable. A pharmaceutically acceptable carrier must be of sufficiently high purity and sufficiently low toxicity to be suitable for administration to the mammal being treated. It must also maintain the stability and bioavailability of the nutritional supplement. A pharmaceutically acceptable carrier may be liquid or solid, and is selected to provide the desired bulk, consistency, etc., when combined with the nutritional supplement and other components of a given composition, taking into account the planned mode of administration.

[0251] The term "nutraceutical" or "dietary supplement" as used herein refers to a food that provides a health benefit in addition to its basic nutritional value.

[0252] The term "nutraceutical amount" or "effective amount" of an active agent is used interchangeably to refer to an amount that is sufficient to provide the intended health benefit. Dosage level is based on a variety of factors, including the type of injury, the patient's age, weight, sex, medical condition, severity of the condition, route of administration, and the specific active agent used. Thus, dosage regimens can vary widely, but can be routinely determined by a physician using standard methods.

[0253] The term "nutraceutical effect" as used herein refers to the result or consequence of exposure to an active ingredient that is not intended to diagnose, treat, cure, or prevent any disease.

[0254] The term "nutraceutical loading," as used herein, refers to the ratio of the weight of a nutraceutical in a particle to the total weight of the particle, expressed as a percentage. It reflects the nutraceutical content of the particle.

[0255] The term "optically active" as used herein refers to the ability of many organic compounds to rotate the plane of plane-polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule about its chiral center(s). The prefixes d and l or (+) and (-) are used to indicate the sign of rotation of plane-polarized light by the compound, with (-) or l meaning that the compound is levorotatory. Compounds prefixed with (+) or d are dextrorotatory.

[0256] The term "parenteral" as used herein refers to a route of administration in which an active substance or drug does not pass through the stomach or "gut" and thus does not encounter the first pass effect of the liver. Examples include, but are not limited to, introduction into the body by ear, eye, intranasal, inhalation, topical, suppository, buccal, sublingual, and injection (i.e., administration by injection), including, for example, subcutaneous (i.e., injection under the skin), intramuscular (i.e., injection into a muscle), intravenous (i.e., injection into a vein), intrathecal (i.e., injection into the space around the spinal cord or the subarachnoid space of the brain), intraventricular injection, intracisternal injection, or infusion techniques.

[0257] The term "particle" as used herein refers to very small components, e.g., nanoparticles or microparticles), that may contain, in whole or in part, at least one active agent as described herein. The term "microparticle" is used herein generally to refer to structures including microcapsules, microparticles, nanoparticles, nanocapsules, and nanospheres. The particles may contain the active agent(s) in a core surrounded by a coating. The therapeutic agent(s) may also be dispersed throughout the particle. The active agent(s) may also be adsorbed to the particle. The particles may be of any order of release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, extended release, etc., and combinations thereof. The particles may contain, in addition to the active agent(s), any of these materials routinely used in the pharmaceutical and medical fields, including, but not limited to, erodible, nonerodible, biodegradable, or nonbiodegradable materials, or combinations thereof. The particles may be microcapsules containing the active agent in solution or in a semi-solid state. The particles may be of virtually any shape.

[0258] The term "particle size distribution" as used herein refers to a statistical measure of the percentage of particles of a particular size (or of a particular size interval). An exemplary descriptive way of representing a particle size distribution is a histogram, where the width of the bar corresponds to the lower or upper limit of a size class and the height of the bar corresponds to the amount of that size class. In particle measurement techniques, it is common to generate cumulative distributions from class-dependent values. For this purpose, the amounts of each measurement class are summed starting from the smallest part. This generates a curve that increases continuously from 0% to 100%, i.e. the "cumulative curve". The cumulative particle size distribution is denoted by the letter Q. Each value Q(x) indicates the amount of the sample consisting of particles smaller than size x. Many statistical parameters can be derived from the particle size distribution. The cumulative distribution is particularly suitable for this purpose. Among the most important parameters are the percentiles. These indicate in each case the size x below which a certain amount of the sample lies. The percentiles are represented by the letter d followed by a % value. The mean value (or mean particle size) can also be calculated from the tabulated values. This calculation is done by multiplying the amount of each measurement class by the average size measurement class and summing the individual values. The cumulative curve is also useful for determining oversized and undersized particles, which are a small portion of particles that are significantly larger or smaller than the bulk of the sample.

[0259] The term "partition coefficient" (P) as used herein is the ratio of the concentration of a substance in the lipid to the concentration in the aqueous phase. It is therefore a measure of lipophilicity. The higher the P, the greater the amount of the substance in the lipid phase (typically octanol) and the greater the lipophilicity. The log of P ("logP") is commonly used for convenience. If P<1, logP is negative, and the compound has greater solubility in the aqueous phase. If P>1, logP is positive, and the compound prefers the lipid phase. The term "LogD" as used herein is the logP of a compound at a particular pH (e.g., logD 5.5 According to some embodiments, the log P of beta-hydroxybutyrate is predicted to be in the range of -0.49 to -0.39.

[0260] As used herein, the term "peripheral nervous system" ("PNS") refers to that part of the nervous system outside the brain and spinal cord that connects the CNS to the sensory organs, other organs of the body, muscles, blood vessels, and glands. It includes the 12 cranial nerves, spinal nerves and roots, and autonomic nerves involved in regulating the muscles of the heart muscle, blood vessel walls, and glands.

[0261] The term "pharmaceutical composition" is used herein to refer to a composition used to prevent, reduce the intensity of, cure, or otherwise treat a target condition or disease.

[0262] The term "pharmaceutically acceptable carrier" as used herein refers to any substantially non-toxic carrier that can be conventionally used for the administration of pharmaceuticals in which the synthetic compounds of the present invention described remain stable and bioavailable. A pharmaceutically acceptable carrier must be of sufficiently high purity and sufficiently low toxicity to be suitable for administration to the mammal being treated. It must also maintain the stability and bioavailability of the active agent. A pharmaceutically acceptable carrier may be liquid or solid, and is selected to provide the desired bulk, consistency, etc., when combined with the active agent and other components of a given composition, taking into account the planned mode of administration.

[0263] The term "pharmaceutically acceptable salt" as used herein refers to a salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and commensurate with a reasonable benefit / risk ratio. When used in medicine, the salt should be pharmaceutically acceptable, although non-pharmaceutically acceptable salts may be conveniently used to prepare pharmaceutically acceptable salts thereof. Such salts include, but are not limited to, those prepared from acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, p-toluenesulfonic acid, tartaric acid, citric acid, methanesulfonic acid, formic acid, malonic acid, succinic acid, naphthalene-2-sulfonic acid, and benzenesulfonic acid. Such salts may also be prepared as alkali metal or alkaline earth salts, such as sodium, potassium, or calcium salts of the carboxylic acid group. "Pharmaceutically acceptable salt" means a salt that is suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., within the scope of sound medical judgment, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, PH Stahl et al. describe pharma-ceutically acceptable salts in detail in "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley VCH, Zurich, Switzerland: 2002). Salts may be prepared in situ during the final isolation and purification of the compounds described herein, or separately by reacting the free base functionality with a suitable organic acid.Representative acid addition salts include, but are not limited to, acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate (isethionate), lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, phosphate, glutamate, bicarbonate, p-toluenesulfonate, and undecanoate salts. Basic nitrogen-containing groups may also be quaternized with agents such as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates, such as dimethyl, diethyl, dibutyl, and diamyl sulfates; long chain halides, such as decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides; aryl alkyl halides, such as benzyl and phenethyl bromides. This results in water- or oil-soluble or dispersible products. Examples of acids that can be used to form pharmaceutically acceptable acid addition salts include inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, and organic acids, such as oxalic acid, maleic acid, succinic acid, and citric acid. Basic addition salts may be prepared in situ during the final isolation and purification of the compounds described within the present invention by reacting the carboxylic acid-containing moiety with a suitable base, such as a hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, or with ammonia or an organic primary, secondary, or tertiary amine.Pharmaceutically acceptable salts include, but are not limited to, cations based on alkali metals or alkaline earth metals, such as lithium, sodium, potassium, calcium, magnesium and aluminum salts, as well as non-toxic quaternary ammonia and amine cations, including ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine. Other representative organic amines useful for the formation of base addition salts include ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine, and the like. Pharmaceutically acceptable salts can also be obtained using standard procedures well known in the art, for example, by reacting a sufficiently basic compound, such as an amine, with a suitable acid to obtain a physiologically acceptable anion. Alkali metal (e.g., sodium, potassium, or lithium) or alkaline earth metal (e.g., calcium or magnesium) salts of carboxylic acids can also be made.

[0264] As used herein, the term "plume width" refers to the width of the plume at a given distance (eg, 3 cm) from the spray nozzle.

[0265] The term "polymer" as used herein refers to a large molecule or macromolecule made up of many repeated subunits (monomers) of identical structure. Polymers that have two different repeating units within their chains are called copolymers.

[0266] As used herein, the term "powder" refers to fine dry particles produced by grinding, crushing, or disintegration of a solid substance.

[0267] As used herein, the term "pump" refers to all components of a container closure system that are involved in metering, atomizing, and delivering a formulation to a subject.

[0268] The term "receptor" as used herein refers to a protein that binds to a specific extracellular signal molecule (ligand) and initiates a response within the cell. Cell surface receptors are located within the plasma membrane with their ligand binding sites exposed to the external medium. Intracellular receptors bind to ligands that diffuse across the plasma membrane into the cell.

[0269] The term "reduce" or "reducing" as used herein refers to a decrease, lowering, attenuation, restriction or reduction in degree, intensity, extent, size, amount, density, number or occurrence.

[0270] "Release" and its various grammatical forms refer to dissolution of the active component and diffusion of the dissolved or solubilized species by a combination of the following processes: (1) hydration of the matrix, (2) diffusion of the solution into the matrix, (3) dissolution of the active, and (4) diffusion of the dissolved active out of the matrix.

[0271] As used herein, the term "second messenger - e.g., cyclic AMP [cAMP], diacylglycerol [DAG], and inositol 1,4,5-triphosphate [IP3] - refers to small molecules that initiate and regulate intracellular signaling pathways.

[0272] The terms "soluble" and "solubility" refer to the property of being susceptible to dissolution in a particular fluid (solvent). The term "insoluble" refers to the property of a material having minimal or limited solubility in a specified solvent.

[0273] A "solution" is generally thought of as a homogeneous mixture of two or more substances, often, but not necessarily, a liquid. In a solution, the molecules of the solute (or dissolved substance) are evenly distributed among the molecules of the solvent.

[0274] As used herein, the term "spray" refers to a system of liquid droplets in a fluid continuous phase.

[0275] As used herein, the term "spray angle" refers to the angle of the emitted plume measured from the apex of the spray cone and spray nozzle.

[0276] The terms "subject" or "individual" or "patient" are used interchangeably to refer to members of animal species of mammalian origin, including, but not limited to, mice, rats, cats, goats, sheep, horses, hamsters, pigs, dogs, guinea pigs, rabbits, and primates (such as, for example, monkeys, apes, or humans).

[0277] As used herein, the phrase "subject in need thereof" refers to a subject who: (i) is at risk for traumatic brain injury involving neuroinflammation; (ii) has suffered from traumatic brain injury involving neuroinflammation; or (iii) has suffered from traumatic brain injury involving neuroinflammation, unless the context and usage of the phrase dictates otherwise.

[0278] The term "support" is used herein to mean to hold, provide, maintain, or help.

[0279] As used herein, the term "predisposed" refers to a member of an at-risk population.

[0280] A "suspension" is a dispersion (mixture) of finely divided undissolved substances dispersed in a liquid vehicle; because they are so finely divided and mixed, they do not settle rapidly. In everyday life, the most common suspensions are suspensions of solids in liquids.

[0281] The term "sustained release" (also referred to as "sustained release") is used herein in its conventional sense to refer to an active formulation that provides gradual release of an active agent over an extended period of time, preferably, but not necessarily, resulting in substantially constant blood levels of the active agent over an extended period of time. Alternatively, delayed absorption of a parenterally administered active form is achieved by dissolving or suspending the active agent in an oil vehicle. Non-limiting examples of sustained release biodegradable polymers include polyesters, polyester polyethylene glycol copolymers, polyamino-derived biopolymers, polyanhydrides, hydrogels, polyorthoesters, polyphosphazenes, SAIBs, photopolymerizable biopolymers, protein polymers, collagen, polysaccharides, chitosan, and alginates.

[0282] The term "symptom" as used herein refers to a sign or manifestation of a disorder or disease, particularly when experienced by an individual as an alteration from normal function, sensation, or appearance.

[0283] The term "synthetic" as used herein refers to being made by a chemical process or modified by deliberate human intervention. As used herein, "synthetic" includes products derived from biological fermentation.

[0284] The term "T cell" or "T lymphocyte" as used herein refers to one of two types of antigen-specific lymphocytes responsible for adaptive immune responses, the other being B cells. T cells are responsible for cell-mediated adaptive immune responses. They originate in the bone marrow, but most of their development takes place in the thymus. The highly variable antigen receptor on T cells is called the T cell receptor. Effector T cells perform a variety of functions in the immune response, always acting by interacting with another cell in an antigen-specific manner. Some T cells activate macrophages, some help B cells produce antibodies, and some kill cells infected with viruses and other intracellular pathogens.

[0285] The term "targeted delivery" as used herein refers to a system that directs the active moiety to its targeted body area (organs, cells, and subcellular levels of specific tissues) to overcome certain toxic effects of conventional active delivery, thereby reducing the amount of active agent required for efficacy.

[0286] The term "therapeutic amount" is an amount sufficient to provide the intended benefit of treatment. By considering factors such as efficacy, relative bioavailability, subject weight, severity of adverse side effects, and preferred mode of administration, in combination with the teachings provided herein, an effective prophylactic or therapeutic treatment regimen may be designed that does not cause substantial toxicity, yet is effective in treating a particular subject.

[0287] The term "therapeutic component" as used herein refers to a therapeutically effective dosage (i.e., dose and frequency of administration) that eliminates, reduces, or prevents a particular disease manifestation in a percentage of the population. An example of a commonly used therapeutic component is the ED50, which describes the dose of a particular dosage that is therapeutically effective against a particular disease manifestation in 50% of the population.

[0288] The term "therapeutic effect" as used herein refers to an outcome of a treatment that is deemed desirable and beneficial. Therapeutic effect may include, directly or indirectly, the arrest, reduction, or elimination of a disease manifestation. Therapeutic effect may also include, directly or indirectly, the arrest, reduction, or elimination of the progression of a disease manifestation.

[0289] The term "therapeutic window" as used herein reflects the concentration range that provides efficacy without unacceptable toxicity. The duration of action of a substance is determined by the period of time that the concentration exceeds the minimum effective concentration (MEC). In general, additional doses can be administered to maintain the concentration within the therapeutic window over time.

[0290] The term "treat" or "treating" includes negating, substantially inhibiting, slowing, or reversing the progression of a disease, condition, or disorder, substantially alleviating the clinical or aesthetic symptoms of a condition, substantially preventing the appearance of clinical or aesthetic symptoms of a disease, condition, or disorder, and protecting against harmful or annoying symptoms. Treating further refers to achieving one or more of the following: (a) reducing the severity of the disorder, (b) limiting the onset of symptoms characteristic of the disorder(s) being treated, (c) limiting the worsening of symptoms characteristic of the disorder(s) being treated, (d) limiting the recurrence of the disorder(s) in patients who previously had the disorder(s), and (e) limiting the recurrence of symptoms in patients who were previously asymptomatic for the disorder(s).

[0291] The term "ubiquitin C-terminal hydrolase L1" or "UCHL1" as used herein refers to a neuron-specific cytoplasmic enzyme that is highly enriched in neurons. It is an indicator of acute injury to brain neurons. It is involved in the degradation of unwanted, misfolded, or damaged proteins. Increased CSF and blood concentrations of UCH-L1 are associated with the process of neuronal destruction (loss) and increased blood-brain barrier permeability. After TBI, blood UCH-L1 levels correlate with the severity of injury. [Mondello, S. et al. BMC Neurology (2012) 12: article 85].

[0292] The terms "ULOQ" or "upper limit of quantitation" are used interchangeably to refer to the highest concentration in a calibration curve that can be determined with a given analytical assay with the required precision and accuracy. Values ​​above the ULOQ are above the limit of quantitation (ALQ).

[0293] Embodiment composition According to one aspect, the present disclosure provides a composition comprising an active ingredient encapsulated in a suspension of particles comprising a biodegradable polymer containing an effective amount of the active ingredient, the particles configured to bypass the blood-brain barrier, the active ingredient is synthetic D-beta hydroxybutyrate, and the targeted delivery is to the brain.

[0294] According to some embodiments, the synthetic beta hydroxybutyrate is soluble in water.

[0295] According to some embodiments, the size of the particles is greater than 10 nm in diameter, greater than 15 nm in diameter, greater than 20 nm in diameter, greater than 30 nm in diameter, greater than 40 nm in diameter, greater than 50 nm in diameter, greater than 60 nm in diameter, greater than 70 nm in diameter, greater than 80 nm in diameter, greater than 90 nm in diameter, greater than 100 nm, greater than 200 nm, greater than 300 nm, greater than 400 nm, greater than 500 nm, greater than 600 nm, greater than 700 nm, greater than 800 nm, greater than 900 nm, greater than 1000 nm, greater than 2000 nm, greater than 3000 nm, greater than 4000 nm, greater than 5000 nm, greater than 6000 nm, greater than 7000 nm, greater than 8000 nm, greater than 9000 nm, greater than 10,000 nm, etc. [See, for example, Ohta, S. et al. Investigating the optimum size of nanoparticles for their delivery into the brain assisted by focused ultrasound-induced blood brain barrier opening. Sci. Reports (2020) article 18220].

[0296] According to some embodiments, the synthetic D-beta hydroxybutyrate is dispersed throughout the particle, or the particle is impregnated with the synthetic D-beta hydroxybutyrate, or the particle comprises a matrix and the matrix comprises synthetic D-beta hydroxybutyrate.

[0297] According to some embodiments, a portion of the synthetic D-beta hydroxybutyrate is adsorbed or weakly bound to the surface of the microparticles and contributes to a rapid initial release or burst release.

[0298] According to some embodiments, the particles have release kinetics of any order, including zero order release, first order release, second order release, delayed release, sustained release, immediate release, extended release, or combinations thereof.

[0299] According to some embodiments, release of synthetic D-beta hydroxybutyrate from the particles is via diffusion, erosion, or both.

[0300] Delivery / Administration Route The invention described relates to all routes of administration including intramuscular, subcutaneous, sublingual, intravenous, intraperitoneal, intranasal, intratracheal, topical, intradermal, intramucosal, intracavernous, intrarectal, intraantral, intragastrointestinal, intraductal, intrathecal, intraventricular, intrapulmonary, intraabscess, intraarticular, subpericardial, intraaxillary, intrapleural space, intradermal, buccal, transmucosal, transdermal, via inhalation, and via subcutaneous injection.

[0301] As used herein, administration includes in vivo administration and direct administration to tissues ex vivo. Generally, the compositions may be administered systemically, i.e., orally, bucally, parenterally, or rectally, in single or multiple dosage unit formulations containing non-toxic pharma- ceutically acceptable carriers, adjuvants, and vehicles, as desired.

[0302] According to some embodiments, the delivery route is via oral administration. The compositions of the invention described may be in a form suitable for oral use, such as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. As used herein, the term "oral" or "orally" refers to introduction into the body by the oral cavity, whereby absorption occurs in one or more of the following body regions: oral cavity, stomach, small intestine, lungs (particularly also referred to as inhalation), and small blood vessels under the tongue (particularly also referred to as sublingual). Compositions intended for oral use may be prepared according to any suitable method, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives to provide a pharma-ceutically elegant and palatable preparation. Tablets may contain the active ingredient(s) in admixture with non-toxic pharma-ceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents, such as corn starch or alginic acid; binders, such as starch, gelatin, or acacia, and lubricants, such as magnesium stearate, stearic acid, or talc. The tablets may be uncoated to protect the composition from oxidation or photodegradation or for controlled release, or may be coated by known techniques, for example, to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period of time. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate may be used.

[0303] The compositions of the invention described may also be formulated for oral use as hard gelatin capsules (wherein the active ingredient(s) are mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin) or as soft gelatin capsules (wherein the active ingredient(s) are mixed with water or an oil medium, such as peanut oil, liquid paraffin, or olive oil).

[0304] The compositions of the present invention described can be formulated as aqueous suspensions, in which the active ingredient(s) are mixed with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents can be naturally occurring phosphatides such as lecithin, or condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more colorants, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0305] The compositions of the present invention described can be formulated as oily suspensions by suspending the active ingredient in vegetable oil, such as peanut oil, olive oil, sesame oil, or coconut oil, or mineral oil, such as liquid paraffin.Oily suspensions may contain thickening agents, such as beeswax, hard paraffin, or cetyl alcohol.Sweetening agents, such as those mentioned above, and flavoring agents may be added to provide a palatable oral preparation.These compositions may be preserved by adding antioxidants, such as ascorbic acid.

[0306] The composition of the present invention described may be formulated in the form of dispersible powder and granules suitable for preparing aqueous suspension by adding water.The active ingredient in such powder and granules is provided in admixture with dispersing or wetting agent, suspending agent, and one or more preservatives.Suitable dispersing or wetting agent and suspending agent are exemplified by those already mentioned above.Additional excipients, such as sweeteners, flavoring agents, and coloring agents, may also be present.

[0307] The composition of the present invention may also be in the form of an emulsion. An emulsion is a two-phase system prepared by combining two immiscible liquid carriers, one of which is uniformly distributed by the other and consists of globules with a diameter equal to or greater than that of the largest colloidal particle. The size of the globules is important and should be such that the system achieves maximum stability. Separation of the two phases does not usually occur unless a third substance (emulsifier) ​​is incorporated. Thus, a basic emulsion contains at least three components: two immiscible liquid carriers and an emulsifier, and an active ingredient.

[0308] Most emulsions incorporate an aqueous phase into a non-aqueous phase (or vice versa).

[0309] However, it is possible to prepare emulsions that are essentially non-aqueous, for example, non-aqueous immiscible systems glycerin and olive oil with anionic and cationic surfactants.The composition of the present invention may therefore be in the form of an oil-in-water emulsion.The oil phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin, or a mixture thereof.Suitable emulsifiers may be naturally occurring gums, for example gum acacia or gum tragacanth, naturally occurring phosphatides, for example soybean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the above partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate.The emulsion may also contain sweeteners and flavorings.

[0310] The compositions of the present invention may also be formulated as syrups and elixirs. Syrups and elixirs may be formulated with sweeteners, such as glycerol, propylene glycol, sorbitol, or sucrose. Such formulations may also contain demulcents, preservatives, and flavoring and coloring agents. Demulcents are protective agents used primarily to relieve irritation, especially mucous membranes or frayed tissue. Several chemicals have demulcent properties. These substances include alginates, mucilages, gums, dextrins, starches, certain sugars, and polymeric polyhydric glycols. Others include acacia, agar, benzoin, carbomer, gelatin, glycerin, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, propylene glycol, sodium alginate, tragacanth, hydrogels, and the like.

[0311] For buccal administration, the compositions of the invention described may take the form of tablets or lozenges formulated in conventional manner.

[0312] There are three common methods of tablet preparation: wet granulation, dry granulation, and direct compression. The preparation method and added ingredients are selected to give the tablet formulation the desired physical characteristics that allow for rapid compression of the tablet. After compression, the tablet must have several additional attributes such as appearance, hardness, disintegration ability, suitable dissolution characteristics, and uniformity, which are also influenced by both the preparation method and added materials present in the formulation.

[0313] According to some embodiments, the tablet is a compressed tablet (CT).Compressed tablets are solid dosage forms formed by compression and do not contain special coatings.Generally, they are made from powdered, crystalline, or granular materials, alone or in combination with binders, disintegrants, controlled release polymers, lubricants, diluents, and colorants.

[0314] According to some embodiments, the tablets are sugar-coated tablets. These are compressed tablets containing a sugar coating. Such coatings may be colored and are useful for masking active substances that have objectionable tastes or odors and for protecting materials sensitive to oxidation.

[0315] According to some embodiments, the tablets are film-coated tablets: these compressed tablets are covered with a thin layer or film of a water-soluble material.

[0316] Many polymeric materials that have film-forming properties may be used.

[0317] According to some embodiments, the tablets are enteric-coated tablets: these compressed tablets are coated with a substance that will withstand solution in gastric juices but disintegrate in the intestine.

[0318] According to some embodiments, the tablet is a multiple compressed tablet. These tablets are made by more than one compression cycle. Layered tablets are prepared by compressing additional tablet granulations on a pre-compressed granulation. The operation can be repeated to produce multi-layer tablets of two or three layers. Press-coated tablets (dry-coated) are prepared by feeding a pre-compressed tablet into a specialized tablet press and compressing another layer of granulation around the pre-formed tablet.

[0319] According to some embodiments, the tablet is a controlled release tablet. Compressed tablets can be formulated to slowly release the active agent over an extended period of time. These dosage forms are therefore referred to as sustained or extended release dosage forms.

[0320] According to some embodiments, the tablets are liquid solution tablets. These compressed tablets can be used to prepare solutions or to impart certain characteristics to solutions.

[0321] According to some such embodiments, the tablets are effervescent tablets. In addition to the active substance, these tablets contain sodium bicarbonate and an organic acid, such as tartaric acid or citric acid. In the presence of water, these additives react and release carbon dioxide, which acts as a disintegrant, producing effervescence.

[0322] According to some embodiments, the tablet is a compressed suppository or insert.

[0323] According to some embodiments, the tablets are buccal and / or sublingual tablets, which are small, flat, oval shaped tablets intended for buccal administration and may slowly dissolve or erode by insertion into the cheek pouch.

[0324] According to some embodiments, the tablet is a molded tablet or a tablet powder.

[0325] In some embodiments, the tablet comprises a compressed core comprising at least one component of the formulation described and a film-forming composition. Formulations utilizing film-forming compositions are known to those skilled in the art (see, for example, Remington's Pharmaceutical Sciences, 20th Ed., 2000). Such film-forming compositions may include, for example, but are not limited to, polymers such as cellulose esters, cellulose ethers, and cellulose ester-ether polymers, amphiphilic triblock copolymer surfactants such as ethylene oxide-propylene oxide-ethylene oxide, and solvents such as acetone that form a film on the core. The compressed core may contain a bilayer core comprising an active layer and a push layer.

[0326] According to some embodiments, when delivered orally, an effective amount of exogenous synthetic D-beta hydroxybutyrate is at least 500 mg to 100 g, inclusive, i.e., at least 500 mg, at least 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, 10 g, 11 g, 12 g, 13 g, 14 g, 15 g, 16 g, , 17g, 18g, 19g, 20g, 21g, 22g, 23g, 24g, 25g, 26g, 27g, 28g, 29g, 30g, 31g, 32g, 33g, 34g, 35g, 36g, 37g, 38g, 3 9g, 40g, 41g, 42g, 43g, 44g, 45g, 46g, 47g, 48g, 49g, 50g, 51g, 52g, 53g, 54g, 55g, 56g, 57g, 58g, 59g, 60g, 61g , 62g, 63g, 64g, 65g, 66g, 67g, 68g, 69g, 70g, 71g, 72g, 73g, 74g, 75g, 76g, 77g, 78g, 79g, 80g, 81g, 82g, 83g, 84g, 85g, 86g, 87g, 88g, 89g, 90g, 91g, 92g, 93g, 94g, 95g, 96g, 97g, 98g, 99g, or 100g, or 10g to 50g, inclusive. That is, it may be in the range of at least 10g, 11g, 12g, 13g, 14g, 15g, 16g, 17g, 18g, 19g, 20g, 21g, 22g, 23g, 24g, 25g, 26g, 27g, 28g, 29g, 30g, 31g, 32g, 33g, 34g, 35g, 36g, 3g, 38g, 39g, 40g, 41g, 42g, 43g, 44g, 45g, 46g, 47g, 48g, 49g, 50g.

[0327] Intranasal delivery of the dosage form. According to some embodiments, delivery is targeted to the brain by intranasal delivery.

[0328] FIG. 4 is an illustrative diagram showing the internal nasal anatomical structure in a cross-section. The human nasal cavity is divided into two halves by the nasal septum, a central part of bone and cartilage, and each symmetrical half opens on the face through the nostrils and connects with the mouth at the nasopharynx. The nasal cavity is subdivided into the nasal vestibule, inferior turbinate, middle turbinate, superior turbinate, olfactory region, frontal sinus, paranasal sinuses, and cribriform plate of the ethmoid bone. [Javia, A. et al., Polymers in nasal drug delivery: an overview. Ch. 11 in Applications of Polymers in Drug Delivery, 2d ed. Misra, A. and Shahiwala, A. Eds. Elsevier (2021) pp. 306-323].

[0329] The area of ​​the nasal vestibule is approximately 0.6 cm 2 The anterior portion of the nasal cavity, just inside the nostrils. The sinuses, also known as nasal hairs, are present in this area and act as a filtering unit for inhaled particles. This nasal portion is lined by stratified squamous epithelium and keratinized epithelium with sebaceous glands [Ibid. Marttin, E. et al Nasal mucociliary clearance as a factor in nasal drug delivery. Adv. Drug Deliv. Rev. (1998) 29:13-38; Illum, L. Nasal drug delivery-possibilities, problems and solutions. J. Controlled Re. (2003) 87:187-98]. These features are necessary for resistance to toxic environmental agents but also affect the absorption of active substances [Kimbell, JS et al. Correlation of regional formaldehyde flux predictions with the distribution of formaldehyde-induced squamous metaplasia in F344 rat nasal passages. Mutat. Res. Fund. Mol. Mech. Mutagen (1997) 380:143-54].

[0330] The anterior chamber lies midway between the nasal vestibule and the respiratory area. Its anterior section is stratified squamous epithelium and its posterior region is pseudostratified columnar cells exhibiting microvilli. [Ibid. citing Marttin, E. et al Nasal mucociliary clearance as a factor in nasal drug delivery. Adv. Drug Deliv. Rev. (1998) 29:13-38].

[0331] In humans, the olfactory region (approximately 15 cm 2 ) is located in the superior turbinate opposite the septum just below the cribriform plate of the ethmoid bone, separating the cranial cavity from the nasal cavity and occupying about 5% of the total area of ​​the nasal cavity [Ibid., citing Illum, L. Is nose-to-brain transport of drugs in man a reality? (2004) J. Pharm. Pharmacol. (2004) 56:3-17], but has a highly vascularized epithelial layer, bypasses the BBB, and has connectivity with cerebrospinal fluid (CSF) and the CNS, making it interesting for delivering substances to the CNS [Ibid., Hanson, LR and Frey, WH Intranasal delivery bypasses the blood-brain barrier to target therapeutic agents to the central nervous system and treat neurodegenerative disease. (2008) BMC Neurosci. 9:S5; Illum, L. Nasal drug delivery-possibilities, problems and solutions. J. Controlled [Citing Re. (2003) 87:187-98]

[0332] The blood-brain barrier (BBB) ​​is located at the level of the cerebral microvasculature and is important for maintaining homeostasis of the central nervous system (CNS). The BBB limits the entry of potentially neurotoxic substances into the brain, but also presents a major obstacle to the delivery of therapeutic agents to the CNS for disease treatment. The BBB has a low rate of pinocytosis and tight junctions (TJs) that form a tight seal between opposing endothelial membranes [Lockhead, JJ and Thorne, RG. “Intranasal delivery of biologics to the central nervous system.” Adv. Drug Deliv Revs. (2012) 64:614-28, citing Reese, TS and Karnovsky, MJ. J. Cell Biol. (1967) 34:207-17]. The presence of TJs at the BBB creates a high transcutaneous electrical resistance of 1500-2000 Ω·cm2, compared to 3-30 Ω·cm2 across most peripheral microvessels [Ibid., citing Crone, C. and Olesen, SP. Brain Res. (1982) 241:49-55; Butt, AM et al. J. Physiol (1990) 429:47-62]. This high resistance is associated with a very low permeability, i.e., the BBB severely restricts paracellular diffusion of solutes from the blood to the brain. Typically, only small lipophilic molecules appreciably cross the normal, healthy BBB via transcellular passive diffusion, although some limited transport of certain peptides and peptide analogues has been reported [Ibid., citing Banks, WA. MBMC Neurol. (2009) 9(Suppl.1):S3]. Essential nutrients such as glucose or iron enter and are acquired in the CNS via specific transporters such as glucose transporter 1 or receptors such as the transferrin receptor [Ibid., citing Dick, AP et al. Proc. Natl Acad. Sci. USA (1984) 81:7233-37; Jeffries, WA, et al. Nature (1984) 312:162-3].Receptors and transporters for gastrointestinal hormones involved in regulating metabolism are expressed at the BBB to transmit information between the CNS and the periphery [Ibid., citing Banks, WA. Regul. Pept. (20008) 149:11-14]. In addition to having low paracellular permeability and low rates of pinocytosis, the BBB also expresses numerous drug transporters (e.g., P-glycoprotein) that further limit the entry into the brain of many endogenous and exogenous substances that would otherwise be expected to cross the BBB based on molecular weight (MW) and lipophilicity considerations. [Ibid.]

[0333] The exact pathways and mechanisms by which drugs move from the nasal epithelium to various regions of the CNS are not fully understood. Substances can cross the olfactory membrane by three mechanisms: transport along the primary neurons of the olfactory and trigeminal nerves in the olfactory epithelium and transport to the olfactory bulb by intracellular axonal transport; substances penetrate across the olfactory supporting epithelium cells by either transcellular or paracellular mechanisms and are then taken up into the CNS; and pinocytosis by the olfactory nerve with the possibility of continuous distribution to more distal brain tissues [Kimbell, JS et al. Correlation of regional formaldehyde flux predictions with the distribution of formaldehyde-induced squamous metaplasia in F344 rat nasal passages. Mutat. Res. Fund. Mol. Mech. Mutagen (1997) 380: 143-54, citing Illum, L. Is nose-to-brain transport of drugs in man a reality? (2004) J. Pharm. Pharmacol. (2004) 56: 3-17].

[0334] The olfactory lobe provides a pathway for substances to enter the CNS and peripheral circulation. Substances can enter the CNS by intraneuronal pathways that transport active substances through axons, which take hours to days for the substance to reach different areas of the brain. Extraneuronal pathways transport substances through perineural channels, which take minutes for the substance to reach parenchymal tissue or CSF [Singh,AK et al.Nasal cavity,a promising,a promising transmucosal platform for drug delivery and research approaches from nasal to brain targeting.J.Drug Deliv.Thera.(2012)2]. Thus, nasal delivery has many advantages over other delivery routes, including rapid absorption, rapid action, and low risk of overdosing [Ibid., citing Arora, P. et al. Permeability issues in nasal drug delivery. Drug Discov. Today (2002) 7:967-75; Ugwoke, ML et al. The biopharmaceutical aspects of nasal mucoadhesive drug delivery. J. Pharm. Pharmacol. (2001) 53:3-22; Romeo, V. et al. Effects of physicochemical properties and other factors on systemic nasal drug delivery. Adv. Drug Deliv. Rev. (1998) 29:89.

[0335] The respiratory epithelium is the main lining of the human nasal cavity and is essential for the cleansing of the nasal mucosa by the nasal mucociliary clearance (NMCC). The respiratory surface area is approximately 130 cm 2[Ibid., citing Pires, A, et al. Intranasal drug delivery: how, why and what for? J. Pharm. Pharm. Sci. (2009) 12:288-311]. It is composed of five types of cells: ciliated columnar cells, non-ciliated columnar cells, basal cells (about 80%), neurosecretory cells of the basement membrane, and goblet cells (about 20%). These cells help to prevent the mucosa from drying out by trapping moisture.

[0336] The nasopharynx is located in the posterior region, with ciliated cells in its upper part and squamous epithelium in its lower part. Approximately 20-40 ml of watery mucus is secreted per day by goblet cells and mixed glands [ibid., citing Quraishi, M. et al. The rheology of nasal mucus: a review. Clin. Otolaryngol. Allied Sci. (1998) 23: 403-13]. The nasopharynx consists of the nasal-associated lymphoid tissue [Ibid., citing Illum, L. Nanoparticulate systems for nasal delivery of drugs: a real improvement over simple systems” J. Pharm. Sci. (2007) 96:473-83], which is located just below the nasal mucosa. [Ibid., citing Stanley, A. et al. Characterization of ovine nasal-associated lymphoid tissue and identification of M cells in the overlying follicle-associated epithelium. J. Comp. Pathol. (2001) 125” 262-70]. Substances administered by the nasal route can be absorbed into the systemic circulation or into the brain, or in many cases both. Absorption is controlled by many factors, including the influx of chemicals into the brain, active metabolic enzymes in the olfactory mucosa, the olfactory bulb, and the brain itself.

[0337] Biological barriers to nasal absorption Numerous studies have demonstrated intranasal (IN) drug delivery to effectively treat animal models of CNS disease. Most of these studies did not present pharmacokinetic data clearly indicating brain uptake of nasally administered compounds, but rather presented pharmacodynamic data showing positive effects after IN delivery of substances in animal models. [Lockhead, JJ et al. “Intranasal delivery of biologics to the central nervous system. Adv. Drug Deliv. Rev. (2012) 64 7]: 614-28]. This makes it difficult to ascertain whether the API entered the CNS via a direct nose-to-brain route, bypassing the BBB, or whether it entered the CNS through the BBB, or perhaps exerted its effects by acting on the brain endothelium.

[0338] Mucus. The first step of absorption by the nasal route is passage through the mucous membrane. Lipophilic active substances can easily cross biological membranes via the transcellular route, since they can partition into the lipid (bilayer) of cell membranes. Thus, lipophilic active substances can rapidly enter both the CSF and the bloodstream after nasal administration, while polar active substances can pass through the mucous layer but cannot pass through the lipid layer. [[Javia,A.et al.,Polymers in nasal drug delivery:an overview.Ch.11 in Applications of Polymers in Drug Delivery,2d ed.Misra,A.and Shahiwala,A.Eds.Elsevier(2021)pp.306-323, Kimbell,JS et al.Correlation of regional formaldehyde flux predictions with the distribution of formaldehyde-induced squamous metaplasia in F344 citing rat nasal passages.Mutat.Res.Fund.Mol.Mech.Mutagen(1997)380:143-54, Sakane, T. et al.The transport of a drug to the cerebrospinal fluid directly from the nasal cavity:the relation to the lipophilicity of the Citing drug.Chem.Pharm.Bull.(1991)39:2456-58].

[0339] Nasal mucociliary clearance (NMCC). The normal half-life time of mucociliary transport in humans is approximately 12-15 minutes [ibid., citing Marttin, E. et al Nasal mucociliary clearance as a factor in nasal drug delivery. Adv. Drug Deliv. Rev. (1998) 29:13-38]. The main components involved in nasal mucociliary clearance (NMCC) are the density of the ciliary body, the periciliary fluid, and the composition of the mucus. The rate of NMCC is highly variable in different regions and under different physiological conditions. Mucociliary activity is regulated by several factors such as temperature, intracellular calcium and cyclic adenosine monophosphate (cAMP) levels, as well as by extracellular ATP, age, sex, posture, sleep, exercise, and common environmental pollutants that can affect ciliary beat frequency and mucus production, resulting in an increased mucociliary clearance rate. Disease states may also affect NMCC [Ibid., citing Turker, S. et al. Nasal route and drug delivery systems. Pharm World Sci. (2004) 26:137-42].

[0340] The absorption of a substance by the nasal mucosa depends on the period of time the substance remains in contact with the nasal mucosa. Thus, impairment in NMCC limits active absorption. To maximize active substance transport to the brain, NMCC needs to be increased to ensure extended contact time of the active substance at the absorption site [ibid., citing Charlton, S. et al Evaluation of bioadhesive polymers as delivery systems for nose to brain delivery: in vitro characterization studies. J. Controlled Rel. (2007) 118: 225-34]. Efforts made to increase the contact / residence time of formulations in the nasal mucosa include polymer properties such as mucoadhesion, pH sensitivity, and thermal gelation. [Ibid., citing Schaefer,ML et al. Trigeminal collaterals in the nasal epithelium and olfactory bulb: a potential route for direct modulation of olfactory information by trigeminal stimuli. J.comp.Neurol.(2002)444:221-26]

[0341] Enzyme Barrier. The enzymatic barrier of the nasal mucosa is primarily responsible for the reduced bioavailability of peptides and proteins across the nasal mucosa. This barrier can be overcome by using enzyme inhibitors, absorption enhancers (e.g., bile salts, cyclodextrins, glycocholates), and peptide / protease inhibitors.

[0342] P-glycoprotein efflux transporter. P-glycoprotein (P-gp) is a glycosylated membrane protein present in human nasal respiratory mucosa. [Ibid., citing Wioland, MA et al. CFTR, MDR1 and MRP1 immunolocalization in normal human nasal respiratory mucosa. J. Histochem. Cytochem. (2000) 48:1215-22]. Co-administration of P-gp inhibitors as part of a nasal formulation increases the permeability of nasally administered active agents [Graff, CL and Pollack, GM. P-glycoprotein attenuates brain uptake of substrates after nasal instillation. Pharm. Res. (2003) 20:1225-30].

[0343] Physicochemical characteristics of the active substance. The physicochemical characteristics of the active ingredient, including molecular weight, solubility, dissolution rate, charge, partition coefficient, acid dissociation constant (pKa), particle size and polymorphism, affect active absorption.

[0344] Substances with molecular weights less than 300 Da easily permeate the aqueous channels of the membrane. For compounds with molecular weights greater than 300 Da, the permeation rate is significantly affected. [Yamamoto, A. et al Absorption of water-soluble compounds with different molecular weights and [Asu1.7]-eel calcitonin from various mucosal administration sites. J. Controlled Rel. (2001) 76: 363-74].

[0345] Biological membrane permeability is primarily influenced by its lipophilicity [Ibid., citing Corbo, DC et al. Characterization of the barrier properties of mucosal membranes. J. Pharm. Sci. (1990) 79:202-6], and low molecular weight lipophilic active substances are absorbed very efficiently through the nasal epithelium [Ibid., citing Hinchcliffe, M. and Illum, L. Intranasal insulin delivery and therapy. Adv. Drug Deliv. Rev. (1999) 35:199-234].

[0346] Particles of different sizes accumulate in different regions of the respiratory tract, for example particles larger than 10 μm accumulate in the respiratory region through breathing, particles smaller than 5 μm are inhaled and reach the lungs, and particles smaller than 0.5 μm are exhaled [Sanders, P. et al. Inhalers in healthy subjects and asthmatic patients. STP Pharma Sci. (1997) 7:3000-76].

[0347] Dissolution is another rate-limiting step of nasal absorption, since nasal absorption occurs only after dissolution of the active substance.Therefore, for nasal powder and suspension dosage forms, dissolution rate is an important parameter.After nasal administration, if dissolution is slow, particles are removed from the airways with subsequent reduced bioavailability [Romeo, V. et al Effects of physicochemical properties and other factors on systemic nasal drug delivery.Adv.Drug Deliv.Rev.(1998)29:39].

[0348] Physicochemical properties of the formulation.

[0349] pH: Nasal absorption and penetration of active substances are often affected by the pKa value and pH of the formulation, so the general rule is that the pH should be selected in the range of 4.5 to 6.5. [Javia, A. et al., Polymers in nasal drug delivery: an overview. Ch. 11 in Applications of Polymers in Drug Delivery, 2d ed. Misra, A. and Shahiwala, A. Eds. Elsevier (2021) pp. 306-323].

[0350] Viscosity: Increasing the viscosity of the formulation may prolong the contact / residence time of the formulation in the nasal cavity, which may increase penetration, but highly viscous formulations may alter nasal regulation functions, including mucociliary clearance and / or ciliary beat frequency. [Ibid.]

[0351] Osmolarity. More often than not, isotonic solutions / preparations are administered to the nasal cavity due to shrinkage of the nasal epithelium in order to increase the permeability of substances due to structural changes. Isotonic solutions also inhibit ciliary activity. {Ibid.}

[0352] Buffer capacity. Acceptable buffer capacity of the formulation may be required to maintain pH. [Ibid.]

[0353] Active concentration, dose, dose volume. The higher the active concentration, the higher the absorption / permeation. The upper limit is 25 mg / dose. Dose volume is ideally in the range of 0.05-0.15 mL / dose (inclusive). [Ibid.]

[0354] Polymers. Polymers such as cellulose derivatives, polyacrylates, starch, gelatin, phospholipids, chitosan, poly-N-alkylacrylamides / poly-N-isopropylacrylamides, cyclodextrins, poloxamers, and methylcellulose have proven effective in improving intranasal absorption of therapeutic agents. [Ibid.]

[0355] Starch microspheres. Bioadhesive starch microspheres absorb water from the nasal mucosa and aid in paracellular transport. [Id., citing Illum, L. et al. Bioadhesive starch microspheres and absorption enhancing agents act synergistically to enhance the nasal absorption of polypeptides. Int. J. Pharm. (2001) 222:109-19].

[0356] Cyclodextrins. Active agent solubility can be increased by encapsulation in carriers such as cyclodextrins. Intranasal delivery along the olfactory pathway to the CNS can be specifically enhanced by using meta-cyclodextrins. [Ibid. citing Marttin, E. et al. Efficacy, safety and mechanism of cyclodextrins as absorption enhancers in nasal delivery of peptide and protein drugs. J. Drug Target (1998) 6:17-36; Merkus, F. et al. Cyclodextrins in nasal drug delivery. Adv. Drug Deliv. Rev. (1999) 36:41-57].

[0357] The shortcomings of solution dosage forms plagued by NMCC can be overcome by the combination of a mucoadhesive polymer and a thermoreversible polymer.

[0358] Thermoresponsive polymers: Thermoresponsive polymers have a hydrophobic-hydrophilic balance in their structure such that they respond to small changes in temperature near a critical temperature by either chain collapse or chain expansion. Examples of thermoreversible polymers include gelatin, carrageenan, methylcellulose, hydroxypropylmethylcellulose (HPMC), xyloglucan, poly(N-isopropylacrylamide-c-acrylic acid), poly(N-isopropylacrylamide (PNIPAAm) / polyethylene oxide (PEO), poloxamer (Pluronic), and PEO / polylactic-co-glycolic acid (PLGA). [Ibid., citing Sharma, N. et al. Mucoadhesive thermoreversible nasal delivery system. J. Pharm. Res. (2010) 3:991-97]. Factors that affect the thermoreversible properties of the formulation are the concentration of the polymer, molecular weight, transition temperature, hydration value, polymer morphology, crystalline state, and polymorphism of the polymer, as well as phase separation of the polymer. [Ibid., Wanka, G. et al. The aggregation behavior of poly-(oxyethylene)-poly-(oxypropylene)-poly-(oxyethylene)block copolymers in aqueous solution. Colloid Polym. Sci. (1990) 268:101-17]. Methylcellulose and HPMC form gels due to hydrophobic interactions between methoxyl substituents. At lower temperatures there is little polymer-polymer interaction, and as temperature increases the polymer loses water of hydration and becomes a gel with low viscosity. After adequate dehydration of the polymers, polymer-polymer association occurs and the entire polymer structure functions as an infinite gel network.

[0359] pH-sensitive polymers. Polymers that are pH-sensitive have a low viscosity under normal ambient conditions, but gel upon nasal administration as a result of changes in pH in the local nasal mucosa. [Ibid., citing Shaikh, RP et al. A review of multi-responsive membranous systems for rate-modulated drug delivery. AAPS PharmaSciTech. (2010) 11:441-59]. This enhances nasal residence time and nasal absorption. These polymers are also described as polyelectrolytes, containing ionizable weak acid or basic moieties attached to a hydrophobic backbone in their structure. Polyacidic polymers with acidic groups remain unswollen at low pH because the acidic groups of the polymer are in protonated, non-ionized form. Negatively charged polymers swell when the pH of the polymer environment increases. In contrast, polybasic polymers swell at lower pH because the basic groups of the polymer are in ionized form. Examples of weak polyacidic pH-sensitive anionic polymers are polycarboxylic acids, such as polyacrylic acid (PAA) or polymethacrylic acid, and polysulfonamides [Ibid., citing Park, SY and Bae, YH, Novel pH-sensitive polymers containing sulfonamide groups. Macromol. Rapid Commun. (1999) 20:269-73]. Depending on the electron-absorbing properties of the substituents on the nitrogen, the pKa of weak polyacids varies between 3 and 11. Examples of cationic pH-sensitive polyelectrolytes include poly(N,N-dialkylaminoethyl methacrylate), polylysine, polyethyleneimine, and chitosan.

[0360] Mucoadhesive polymers. Nasal absorption of substances can be improved by prolonging the retention of particles in the nasal mucosa. In nasal active delivery, mucoadhesion involves the interaction between mucus secreted by the submucosal glands and mucoadhesive polymers [ibid., citing Salamat-Miller, N. et al. The use of mucoadhesive polymers in buccal drug delivery. Adv. Drug Deliv. Rev. (2005) 57:1666-91]. The successive steps of mucoadhesion include (1) absorption of water from the mucous layer leading to wetting and swelling of the polymer, (2) upon swelling of the polymer, hydrogen bonds between the polymer chains dissociate, causing a decrease in polymer-polymer interactions and an increase in polymer-water interactions, after which the free polymer chains penetrate the mucus [Ibid., citing Ugwoke, ML et al Nasal mucoadhesive drug delivery: background, applications, trends and future perspectives. Adv. Drug Deliv. Rev. (2005) 57:1640-65] and then interact with protein chains present in the nasal mucosa. Thus, the formulation is retained in the nasal cavity, resulting in an increased active agent concentration gradient across the epithelium. [Ibid., citing Jimenez-Castellanos, MR et al. Mucoadhesive drug delivery systems. Drug. Dev. Ind. Pharm. (1993) 19:143-94]. Exemplary mucoadhesive polymers include chitosan, alginate, and cellulose or its derivatives.

[0361] Ion-Responsive Polymers. In this category, the sol-gel transition is due to the presence of monovalent / divalent cations such as Na+, K+, Ca2+ and Mg2+. The most judiciously used polymers in this category are the naturally occurring anionic polymers. Gellan gum, carrageenan, pectin, sodium alginate and xyloglucan have the property of cation-induced gelation. [Ibid.]

[0362] According to some embodiments, for intranasal delivery, the maximum amount is administered every 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 hours, 74 hours, 75 hours, 76 hours, 77 hours, 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 83 hours, 84 hours, 85 hours, 86 hours, 87 hours, 88 hours, 89 hours, 90 hours, 91 hours, 92 hours, 93 hours, 94 hours, 95 hours, 96 hours, 97 hours, 98 hours, 99 hours, 100 hours, 0 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 3;8 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, etc.

[0363] According to some embodiments, based on the HED from the mouse study reported in the Example at paragraph

[0634] below, the minimum daily dose for a 60 kg human (132 lbs) is 250 mg / day. According to some embodiments, the minimum daily dose for a 90 kg human (200 lbs) is 730 mg / day.

[0364] According to some embodiments, the dose-time curve is determined to be at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, or at least 24 hours, for at least one day, at least every 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes, as tolerated. A maximum of 25 mg / dose delivered intranasally for at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc. is determined to achieve the desired effective amount.

[0365] According to some embodiments, the subject is administered within 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 14 days, 15 days, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 18 days, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 24 hours, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 hours, 45 minutes, 46 minutes, 47 minutes, 48 ​​minutes, 49 minutes, 50 minutes, 51 minutes, 52 minutes, 53 minutes, 54 minutes, 55 minutes, 56 hours, 57 hours, Intranasal delivery for 3 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, and for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., after participating in an event where acquired brain injury is a known risk is neuroprotective. According to some embodiments, the event is a military operation. According to some embodiments, the event is a sporting event.According to some embodiments, the sporting event is, for example, hockey, football, soccer, baseball, polo, rugby, horse riding, auto racing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, bicycle motocross (BMX), motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing. Other uses include emergency medical care, for example, by EMTs or emergency departments in the context of such emergency situations, such as domestic violence, falls, car accidents, etc.

[0366] According to some embodiments, the incorporation of D-beta hydroxybutyrate into a polymeric carrier can achieve controlled release, including delayed release and sustained release.

[0367] According to some embodiments, D-beta hydroxybutyrate binds to HCA2.

[0368] According to some embodiments, the composition, when delivered intranasally, increases brain D-beta hydroxybutyrate levels for, e.g., at least 6 hours to at least 48 hours, inclusive, i.e., at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, at least 24 hours, at least The concentration may be increased to about 0.1-3 mmol / L, or to about 0.1-1 mmol / L, for at least 25 hours, at least 26 hours, at least 27 hours, at least 28 hours, at least 29 hours, at least 30 hours, at least 31 hours, at least 32 hours, at least 33 hours, at least 34 hours, at least 35 hours, at least 36 hours, at least 37 hours, at least 38 hours, at least 39 hours, at least 40 hours, at least 41 hours, at least 42 hours, at least 43 hours, at least 44 hours, at least 45 hours, at least 46 hours, at least 47 hours, or at least 48 hours.

[0369] According to another aspect, the disclosure provides a method for promoting brain healing following traumatic brain injury including neuroinflammation, comprising: formulating a composition comprising an active ingredient as a suspension of particles comprising a biodegradable polymer containing an effective amount of the active ingredient; administering to a subject the composition comprising the active ingredient, wherein the active ingredient is D-beta hydroxybutyrate; and targeting the composition to the brain, wherein the healing comprises reducing neuroinflammation.

[0370] According to some embodiments, the administering is oral or intranasal.

[0371] According to some embodiments, when delivered orally, an effective amount of exogenous synthetic D-beta hydroxybutyrate is at least 500 mg to 100 g, inclusive, i.e., at least 500 mg, at least 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, 10 g, 11 g, 12 g, 13 g, 14 g, 15 g, 16 g, 17 g, 18 g, 19 g, 20 g, 21 g, 22 g, 23 g, 24 g, 25 g, 26 g, 27 g, 28 g, 29 g, 30 g, 31 g, 32 g, 33 g, 34 g, 35 g, 36 g, 37 g, 38 g, 39 g, 40 g, 41 g, 42 g, 43 g, 44 g, 45 g, 46 g, 47 g, 48 g, 49 g, 50 g, 51 g, 52 g, 53 g, 54 g, 55 g, 56 g, 57 g, 58 g, 59 g, 60 g, 61 g, 62 g, 63 g, 64 g, 65 g, 66 g, 67 g, 68 g, 69 g, 70 g, 71 g, 72 g, 73 g, 74 g, 75 g, 76 g, 77 g, 78 g, 79 g, 80 g, 81 g, 82 g, 83 g, 84 g, 85 g, 86 g, g, 17g, 18g, 19g, 20g, 21g, 22g, 23g, 24g, 25g, 26g, 27g, 28g, 29g, 30g, 31g, 32g, 33g, 34g, 35g, 36g, 37g, 38g , 39g, 40g, 41g, 42g, 43g, 44g, 45g, 46g, 47g, 48g, 49g, 50g, 51g, 52g, 53g, 54g, 55g, 56g, 57g, 58g, 59g, 60g, 6 1g, 62g, 63g, 64g, 65g, 66g, 67g, 68g, 69g, 70g, 71g, 72g, 73g, 74g, 75g, 76g, 77g, 78g, 79g, 80g, 81g, 82g, 83g, 84g, 85g, 86g, 87g, 88g, 89g, 90g, 91g, 92g, 93g, 94g, 95g, 96g, 97g, 98g, 99g, or 100g, or 10g to 5 inclusive 0g, i.e., at least 10g, 11g, 12g, 13g, 14g, 15g, 16g, 17g, 18g, 19g, 20g, 21g, 22g, 23g, 24g, 25g, 26g, 27g, 28g, 29g, 30g, 31g, 32g, 33g, 34g, 35g, 36g, 3g, 38g, 39g, 40g, 41g, 42g, 43g, 44g, 45g, 46g, 47g, 48g, 49g, 50g.

[0372] According to some embodiments, for intranasal delivery, the minimum effective amount may be administered at any frequency as tolerated, e.g., every 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 hours, 74 hours, 75 hours, 76 hours, 77 hours, 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 83 hours, 84 hours, 85 hours, 86 hours, 87 hours, 88 hours, 89 hours, 90 hours,and the like for 8 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, etc., inclusive, at a dose of about 85 mg / day (e.g., for a child weighing about 45 pounds) to about 800 mg / day (e.g., for an adult In the case of 25mg / day, about 335mg / day, about 345mg / day, about 355mg / day, about 365mg / day, about 375mg / day, about 385mg / day, about 395mg / day, about 405mg / day, about 415mg / day, about 425mg / day, about 435mg / day, about 445mg / day, about 455mg / day, about 465mg / day, about 475mg / day, about 485mg / day, about 495mg / day, about 505mg / day, about 515mg / day, about 525mg / day, about 535mg / day, about 545mg / day, about 555mg / day, about 565mg / day, about 575mg / day, about 5 a maximum of about 25 mg / dose to achieve a minimum daily dose of 85 mg / day, about 595 mg / day, about 605 mg / day, about 615 mg / day, about 625 mg / day, about 635 mg / day, about 645 mg / day, about 655 mg / day, about 665 mg / day, about 675 mg / day, about 685 mg / day, about 695 mg / day, about 705 mg / day, about 715 mg / day, about 725 mg / day, about 735 mg / day, about 745 mg / day, about 755 mg / day, about 765 mg / day, about 775 mg / day, about 785 mg / day, about 795 mg / day, or about 800 mg / day;

[0373] According to some embodiments, based on the human equivalent dose (HED) from the mouse study reported in paragraph

[0634] below, the minimum daily dose for a 60 kg human (132 lbs) is 250 mg / day. According to some embodiments, the minimum daily dose for a 90 kg human (200 lbs) is 730 mg / day.

[0374] According to some embodiments, the dose-time curve is determined such that a maximum of 25 mg / dose delivered intranasally at a given frequency as tolerated, e.g., every 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes, for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., achieves the desired effective amount.

[0375] According to some embodiments, for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, or every 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes prior to participating in an event where acquired brain injury is a known risk. Intranasal delivery is neuroprotective for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., following participation in an event where acquired brain injury is a known risk. According to some embodiments, the event is a military operation. According to some embodiments, the event is a sporting event.According to some embodiments, the sporting event may be, for example, hockey, football, soccer, baseball, polo, rugby, horse riding, auto racing, motorcycling, skiing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing. Other applications include emergency medical use, such as by EMTs or emergency departments in the context of such emergency situations as domestic violence, falls, motor vehicle accidents, and the like.

[0376] According to some embodiments, the composition, when delivered intranasally, increases brain D-beta hydroxybutyrate levels for at least 6 hours to at least 48 hours, inclusive, i.e., at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, at least 24 hours, at least 25 hours, at least at least 26 hours, at least 27 hours, at least 28 hours, at least 29 hours, at least 30 hours, at least 31 hours, at least 32 hours, at least 33 hours, at least 34 hours, at least 35 hours, at least 36 hours, at least 37 hours, at least 38 hours, at least 39 hours, at least 40 hours, at least 41 hours, at least 42 hours, at least 43 hours, at least 44 hours, at least 45 hours, at least 46 hours, at least 47 hours, or at least 48 hours, inclusive.

[0377] According to some embodiments, the beta hydroxybutyrate is soluble in water.

[0378] According to some embodiments, the size of the particles is greater than 10 nm in diameter, greater than 15 nm in diameter, greater than 20 nm in diameter, greater than 30 nm in diameter, greater than 40 nm in diameter, greater than 50 nm in diameter, greater than 60 nm in diameter, greater than 70 nm in diameter, greater than 80 nm in diameter, greater than 90 nm in diameter, greater than 100 nm, greater than 200 nm, greater than 300 nm, greater than 400 nm, greater than 500 nm, greater than 600 nm, greater than 700 nm, greater than 800 nm, greater than 900 nm, greater than 1000 nm, greater than 2000 nm, greater than 3000 nm, greater than 4000 nm, greater than 5000 nm, greater than 6000 nm, greater than 7000 nm, greater than 8000 nm, greater than 9000 nm, greater than 10,000 nm, etc. [See, for example, Ohta, S. et al. Investigating the optimum size of nanoparticles for their delivery into the brain assisted by focused ultrasound-induced blood brain barrier opening. Sci. Reports (2020) article 18220].

[0379] According to some embodiments, the D-beta hydroxybutyrate is dispersed throughout the particle, or the particle is impregnated with the D-beta hydroxybutyrate, or the particle comprises a matrix, and the matrix comprises the D-beta hydroxybutyrate.

[0380] According to some embodiments, a portion of the D-beta hydroxybutyrate is adsorbed or weakly bound to the surface of the microparticles and contributes to a rapid initial release or burst release.

[0381] According to some embodiments, the particles have release kinetics of any order, including zero order release, first order release, second order release, delayed release, sustained release, immediate release, extended release, or combinations thereof.

[0382] According to some embodiments, release of D-beta hydroxybutyrate from the particles is via diffusion, erosion, or both.

[0383] According to another aspect, the present disclosure provides a method for promoting brain health, comprising: formulating a composition comprising a suspension of particles comprising a biodegradable polymer containing an effective amount of a drug; administering to a subject a composition comprising an active ingredient, wherein the drug is synthetic D-beta hydroxybutyrate; and targeting the composition to the brain, wherein burst and controlled release of the synthetic D-beta hydroxybutyrate can lead to effective levels of D-beta hydroxybutyrate in the brain.

[0384] According to some embodiments, the administering is oral or intranasal.

[0385] According to some embodiments, when delivered orally, an effective amount of exogenous synthetic D-beta hydroxybutyrate is at least 500 mg to 100 g, inclusive, i.e., at least 500 mg, at least 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, 10 g, 11 g, 12 g, 13 g, 14 g, 15 g, 16 g, 17g, 18g, 19g, 20g, 21g, 22g, 23g, 24g, 25g, 26g, 27g, 28g, 29g, 30g, 31g, 32g, 33g, 34g, 35g, 36g, 37g, 38g, 39 g, 40g, 41g, 42g, 43g, 44g, 45g, 46g, 47g, 48g, 49g, 50g, 51g, 52g, 53g, 54g, 55g, 56g, 57g, 58g, 59g, 60g, 61g, 6 2g, 63g, 64g, 65g, 66g, 67g, 68g, 69g, 70g, 71g, 72g, 73g, 74g, 75g, 76g, 77g, 78g, 79g, 80g, 81g, 82g, 83g, 84g, 85g, 86g, 87g, 88g, 89g, 90g, 91g, 92g, 93g, 94g, 95g, 96g, 97g, 98g, 99g, or 100g, or a value between 10g and 50g, inclusive; , at least 10g, 11g, 12g, 13g, 14g, 15g, 16g, 17g, 18g, 19g, 20g, 21g, 22g, 23g, 24g, 25g, 26g, 27g, 28g, 29g, 30g, 31g, 32g, 33g, 34g, 35g, 36g, 3g, 38g, 39g, 40g, 41g, 42g, 43g, 44g, 45g, 46g, 47g, 48g, 49g, or 50g.

[0386] According to some embodiments, when delivered intranasally, the maximum dose of 25 mg / dose is administered every 2, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 120, 130, 140, 141, 142, 143, 144, 145, 146, 1and the like for 8 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, or 72 hours, inclusive, at a dose of about 85 mg / day (e.g., for a child weighing about 45 pounds) to about 800 mg / day (e.g., for a child weighing about 45 pounds). For example, for adults, i.e., at least about 85 mg / day, about 95 mg / day, about 105 mg / day, about 115 mg / day, about 125 mg / day, about 135 mg / day, about 145 mg / day, about 155 mg / day, about 165 mg / day, about 175 mg / day, about 185 mg / day, about 195 mg / day, about 205 mg / day, about 215 mg / day, about 225 mg / day, about 235 mg / day, about 245 mg / day, about 255 mg / day, about 265 mg / day, about 275 mg / day, about 285 mg / day, about 295 mg / day, about 305 mg / day, about 31 5mg / day, about 325mg / day, about 335mg / day, about 345mg / day, about 355mg / day, about 365mg / day, about 375mg / day, about 385mg / day, about 395mg / day, about 405mg / day, about 415mg / day, about 425mg / day, about 435mg / day, about 445mg / day, about 455mg / day, about 465mg / day, about 475mg / day, about 485mg / day, about 495mg / day, about 505mg / day, about 515mg / day, about 525mg / day, about 535mg / day, about 545mg / day, about 555mg / day, about 565mg / day , achieving a minimum daily dose of about 575 mg / day, about 585 mg / day, about 595 mg / day, about 605 mg / day, about 615 mg / day, about 625 mg / day, about 635 mg / day, about 645 mg / day, about 655 mg / day, about 665 mg / day, about 675 mg / day, about 685 mg / day, about 695 mg / day, about 705 mg / day, about 715 mg / day, about 725 mg / day, about 735 mg / day, about 745 mg / day, about 755 mg / day, about 765 mg / day, about 775 mg / day, about 785 mg / day, about 795 mg / day, or about 800 mg / day;

[0387] According to some embodiments, based on the HED from the mouse study reported in paragraph

[0634] below, the minimum daily dose for a 60 kg human (132 lbs) is 250 mg / day. According to some embodiments, the minimum daily dose for a 90 kg human (200 lbs) is 730 mg / day.

[0388] According to some embodiments, a dose-time curve is determined such that a maximum of 25 mg / dose delivered intranasally at a given frequency as tolerated, e.g., every 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes, for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., achieves a desired effective amount.

[0389] According to some embodiments, the subject is administered within 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 14 days, 15 days, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 18 days, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 24 hours, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 hours, 45 minutes, 46 minutes, 47 minutes, 48 ​​minutes, 49 minutes, 50 minutes, 51 minutes, 52 minutes, 53 minutes, 54 minutes, 55 minutes, 56 hours, 57 hours, Intranasal delivery for 3 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, and for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., after participating in an event where acquired brain injury is a known risk is neuroprotective. According to some embodiments, the event is a military operation. According to some embodiments, the event is a sporting event.According to some embodiments, the sporting event may be, for example, hockey, football, soccer, baseball, polo, rugby, horse riding, auto racing, motorcycling, skiing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing. Other applications include emergency medical use, such as by EMTs or emergency departments in the context of such emergency situations as domestic violence, falls, motor vehicle accidents, and the like.

[0390] According to some embodiments, the composition, when delivered intranasally, increases brain D-beta hydroxybutyrate levels for at least 6 hours to at least 48 hours, inclusive, i.e., at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, at least 24 hours, at least 25 hours, at least at least 26 hours, at least 27 hours, at least 28 hours, at least 29 hours, at least 30 hours, at least 31 hours, at least 32 hours, at least 33 hours, at least 34 hours, at least 35 hours, at least 36 hours, at least 37 hours, at least 38 hours, at least 39 hours, at least 40 hours, at least 41 hours, at least 42 hours, at least 43 hours, at least 44 hours, at least 45 hours, at least 46 hours, at least 47 hours, or at least 48 hours, inclusive.

[0391] According to some embodiments, the size of the particles is greater than 10 nm in diameter, greater than 15 nm in diameter, greater than 20 nm in diameter, greater than 30 nm in diameter, greater than 40 nm in diameter, greater than 50 nm in diameter, greater than 60 nm in diameter, greater than 70 nm in diameter, greater than 80 nm in diameter, greater than 90 nm in diameter, greater than 100 nm, greater than 200 nm, greater than 300 nm, greater than 400 nm, greater than 500 nm, greater than 600 nm, greater than 700 nm, greater than 800 nm, greater than 900 nm, greater than 1000 nm, greater than 2000 nm, greater than 3000 nm, greater than 4000 nm, greater than 5000 nm, greater than 6000 nm, greater than 7000 nm, greater than 8000 nm, greater than 9000 nm, greater than 10,000 nm, etc. [See, for example, Ohta, S. et al. Investigating the optimum size of nanoparticles for their delivery into the brain assisted by focused ultrasound-induced blood brain barrier opening. Sci. Reports (2020) article 18220].

[0392] According to some embodiments, the D-beta hydroxybutyrate is dispersed throughout the particle, or the particle is impregnated with the D-beta hydroxybutyrate, or the particle comprises a matrix, and the matrix comprises the D-beta hydroxybutyrate.

[0393] According to some embodiments, a portion of the D-beta hydroxybutyrate is adsorbed or weakly bound to the surface of the microparticles and contributes to a rapid initial release or burst release.

[0394] According to some embodiments, the particles have release kinetics of any order, including zero order release, first order release, second order release, delayed release, sustained release, immediate release, extended release, or combinations thereof.

[0395] According to some embodiments, release of D-beta hydroxybutyrate from the particles is via diffusion, erosion, or both.

[0396] Pharmaceutical Compositions According to another aspect, the present disclosure provides a pharmaceutical composition comprising a pharma- ceutically acceptable carrier and a therapeutic amount of an API, wherein the API is synthetic D-beta hydroxybutyrate (BHB), and wherein the targeted delivery is to the olfactory region of the nasal cavity, wherein the targeted delivery achieves a therapeutically effective amount of the API in the brain.

[0397] According to some embodiments, the pharmaceutical composition comprises an active agent formulated as a solution, suspension or dispersion. According to some embodiments, the API is formulated as a solution. According to some embodiments, the API is formulated as a suspension. According to some embodiments, the API is formulated as a dispersion, the dispersion comprising a particle size distribution of a population of particles comprising the API. According to some embodiments, the API is formulated as a suspension. According to some embodiments, the API is formulated as a dispersion, the dispersion comprising a particle size distribution of a population of particles comprising the API.

[0398] According to some embodiments, the pharmaceutical composition comprises a liquid spray formulation or a dry powder formulation.

[0399] According to some embodiments, the pharmaceutical composition comprises a liquid spray formulation of the API. According to some embodiments, the pharmaceutical composition is delivered as a liquid spray comprising a droplet size distribution comprising droplets containing the API.

[0400] According to some embodiments, the pharmaceutical composition comprises a dry powder formulation. According to some embodiments, the dry powder formulation when aerosolized comprises a cloud of very fine particles comprising the API.

[0401] According to some embodiments, the formulation is formulated with an excipient. According to some embodiments, the excipient maximizes the solubility of the API. According to some embodiments, the excipient enhances the adhesion of the API to the nasal mucosa in the olfactory region. According to some embodiments, the enhanced adhesion enhances the absorption of the API in the nasal mucosa.

[0402] According to some embodiments, the nasal aerosol comprises the formulation, the container, the valve, the actuator, the dust cap, the associated accessories, and the protective packaging, which together constitute the drug product. For administration by injection, for example, the pharmaceutical composition according to the invention described can be conveniently delivered in the form of an aerosol spray presentation from a pressurized pack using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. For pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges, for example gelatin capsules and cartridges for use in inhalers, can be formulated to contain a powder mix of the active compound and a suitable powder base, for example, lactose or starch.

[0403] According to some embodiments, the formulation is loaded into a unit dose device. According to some devices, the formulation is loaded into a multi-dose device. According to some embodiments, exemplary devices include a spray pump and a metered dose inhaler. According to some embodiments, the liquid formulation is loaded into a spray pump, which is an exemplary device for delivering intranasal formulations. According to some embodiments, the dry powder formulation is loaded into a dry powder device that includes a formulation of active agent and a container closure system.

[0404] For example, a typical nasal spray formulation consists of an active agent suspended or dissolved in an aqueous medium that is loaded into a bottle with a metered spray pump. Actuation of the pump by the subject delivers droplets containing the drug into the nasal cavity. The role of the pump, an integral part of the entire assembly, is to spray and deliver a precise drug dose.

[0405] Most nasal spray pumps produce droplets in the range of about 20 μm to about 120 μm, inclusive, i.e., about 20 μm, about 25 μm, about 30 μm, about 35 μm, about 40 μm, about 45 μm, about 50 μm, 55 μm, about 60 μm, about 65 μm, about 70 μm, about 75 μm, about 80 μm, about 85 μm, about 90 μm, about 95 μm, about 100 μm, about 105 μm, about 110 μm, about 115 μm, or about 120 μm, containing a therapeutic amount of the API [Yu, G. et al. Fluid flow and particle diffusion in the human upper respiratory system. Aerosol Science and Technol. (1998) 28:146-58], such that the droplet size allows the droplets to deposit in the olfactory portion of the nasal cavity for delivery of the API to the brain. If the droplet size is too fine (less than 10 μm), the droplets may pass through the nasal passages and accumulate in the lungs. If the droplet size is too large, the spray may be trapped in the nostrils. Laser diffraction techniques can measure droplet size by measuring the intensity of light scattered by a particle as a function of angle to determine the droplet size for a given device.

[0406] The characteristics of the emitted spray plume are believed to be important in assessing pump performance. According to some embodiments, pump performance is characterized by the spray pattern and plume geometry of the plume emitted by the delivery device. During the very early life of the aqueous nasal spray plume, the formulation may exit the actuator orifice as a thin stream and then form a relatively stable, fully developed cone-shaped plume before separating from the orifice. The term "plume geometry" describes the side view of the aerosol cloud parallel to the axis of the plume. According to some embodiments, the measurement of the plume geometry includes an image taken from the side of the plume perpendicular to the laser light sheet, which is aligned with the expected central plume axis. According to some embodiments, the measurement of the plume geometry is by flash illumination of the plume with photographic image capture. According to some embodiments, the plume geometry is characterized by the spray angle and the plume width.

[0407] According to some embodiments, the pharmaceutical composition comprises a formulation containing a distribution of particles containing the API. According to some embodiments, the majority of the distribution of particles is in the range of about 25 μm to about 40 μm in diameter, inclusive, i.e., about 25 μm, about 26 μm, about 27 μm, about 28 μm, about 29 μm, about 30 μm, about 31 μm, about 32 μm, about 333, about 34 μm, about 35 μm, about 36 μm, about 37 μm, about 38 μm, about 39 μm, or about 40 μm. According to some embodiments, the distribution of particles is depleted of smaller particles that would otherwise enter the lungs.

[0408] According to some embodiments, a viscosity enhancer / mucoadhesive polymer, such as microcrystalline cellulose, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, or a combination thereof, may be added to the formulation at 0.1%, 0.2%, 0.3%, 1% or 2% of the formulation by weight or volume to increase the absorption of the API at the nasal mucosa in the olfactory portion of the nasal cavity. According to some embodiments, the viscosity of each nasal spray formulation is measured. According to some embodiments, the spray pattern and plume geometry of each nasal spray formulation is measured. According to some embodiments, the droplet size distribution of each nasal spray is determined, and the median (Dv50), 10th percentile (Dv10) and 90th percentile (Dv90) of the cumulative undersize distribution are determined.

[0409] According to some embodiments, for intranasal delivery, a maximum of about 25 mg / dose may be administered every 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 hours, 74 hours, 75 hours, 76 hours, 77 hours, 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 83 hours, 84 hours, 85 hours, 86 hours, 87 hours, 88 hours, 89 hours, 90 hours, 91 hours, 92 hours, 93 hours, 94 hours, 95 hours, 96 hours, 97 hours, 98 hours, 99and the like for 8 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, etc., inclusive, ranging from about 85 mg / day (e.g., for a child weighing about 45 pounds) to about 800 mg / day (e.g., for a child weighing about 45 pounds). , for adults), i.e., at least about 85 mg / day, about 95 mg / day, about 105 mg / day, about 115 mg / day, about 125 mg / day, about 135 mg / day, about 145 mg / day, about 155 mg / day, about 165 mg / day, about 175 mg / day, about 185 mg / day, about 195 mg / day, about 205 mg / day, about 215 mg / day, about 225 mg / day, about 235 mg / day, about 245 mg / day, about 255 mg / day, about 265 mg / day, about 275 mg / day, about 285 mg / day, about 295 mg / day, about 305 mg / day, about 315 mg / day, about 325 mg / day, about 335 mg / day, about 345 mg / day, about 355 mg / day, about 365 mg / day, about 375 mg / day, about 385 mg / day, about 395 mg / day, about 405 mg / day, about 415 mg / day, about 425 mg / day, about 435 mg / day, about 445 mg / day, about 455 mg / day, about 465 mg / day, about 475 mg / day, about 485 mg / day, about 495 mg / day, about 505 mg / day, about 515 mg / day, about 525 mg / day, about 535 mg / day, about 545 mg / day, about 555 mg / day, about 565 mg / day, about 575 mg / day, about 585 mg / day, about 595 mg / day, about 605 mg / day, about 615 mg / day, about 625 mg / day, about 635 mg mg / day, about 325 mg / day, about 335 mg / day, about 345 mg / day, about 355 mg / day, about 365 mg / day, about 375 mg / day, about 385 mg / day, about 395 mg / day, about 405 mg / day, about 415 mg / day, about 425 mg / day, about 435 mg / day, about 445 mg / day, about 455 mg / day, about 465 mg / day, about 475 mg / day, about 485 mg / day, about 495 mg / day, about 505 mg / day, about 515 mg / day, about 525 mg / day, about 535 mg / day, about 545 mg / day, about 555 mg / day, about 565 mg / day, achieve a minimum daily dose of about 575 mg / day, about 585 mg / day, about 595 mg / day, about 605 mg / day, about 615 mg / day, about 625 mg / day, about 635 mg / day, about 645 mg / day, about 655 mg / day, about 665 mg / day, about 675 mg / day, about 685 mg / day, about 695 mg / day, about 705 mg / day, about 715 mg / day, about 725 mg / day, about 735 mg / day, about 745 mg / day, about 755 mg / day, about 765 mg / day, about 775 mg / day, about 785 mg / day, about 795 mg / day, or about 800 mg / day;

[0410] According to some embodiments, based on the human equivalent dose (HED) from the mouse study reported in paragraph

[0634] below, the minimum daily dose for a 60 kg human (132 lbs) is 250 mg / day. According to some embodiments, the minimum daily dose for a 90 kg human (200 lbs) is 730 mg / day.

[0411] According to some embodiments, a dose-time curve is determined such that a maximum of 25 mg / dose delivered intranasally at a given frequency as tolerated, e.g., every 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes, for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., achieves a desired effective amount.

[0412] According to some embodiments, the subject is administered within 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 14 days, 15 days, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 18 days, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 24 hours, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 hours, 45 minutes, 46 minutes, 47 minutes, 48 ​​minutes, 49 minutes, 50 minutes, 51 minutes, 52 minutes, 53 minutes, 54 minutes, 55 minutes, 56 hours, 57 hours, Intranasal delivery for 3 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, and for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., after participating in an event where acquired brain injury is a known risk is neuroprotective. According to some embodiments, the event is a military operation. According to some embodiments, the event is a sporting event.According to some embodiments, the sporting event may be, for example, hockey, football, soccer, baseball, polo, rugby, horse riding, auto racing, motorcycling, skiing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing. Other applications include emergency medical use, such as by EMTs or emergency departments in the context of such emergency situations as domestic violence, falls, motor vehicle accidents, and the like.

[0413] According to some embodiments, the incorporation of D-beta hydroxybutyrate into a polymeric carrier can achieve controlled release, including delayed release and sustained release.

[0414] According to some embodiments, D-beta hydroxybutyrate binds to HCA2.

[0415] According to some embodiments, the composition, when delivered intranasally, increases brain D-beta hydroxybutyrate levels for, e.g., at least 6 hours to at least 48 hours, inclusive, i.e., at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, at least 24 hours, at least The concentration may be increased to about 0.1-3 mmol / L, or to about 0.1-1 mmol / L, for at least 25 hours, at least 26 hours, at least 27 hours, at least 28 hours, at least 29 hours, at least 30 hours, at least 31 hours, at least 32 hours, at least 33 hours, at least 34 hours, at least 35 hours, at least 36 hours, at least 37 hours, at least 38 hours, at least 39 hours, at least 40 hours, at least 41 hours, at least 42 hours, at least 43 hours, at least 44 hours, at least 45 hours, at least 46 hours, at least 47 hours, or at least 48 hours.

[0416] method. (1) Before damage

[0417] According to another aspect, the disclosure provides a method for reducing the risk of brain damage due to acquired brain injury in a subject susceptible to or otherwise at risk for acquired brain injury, comprising administering a pharmaceutical composition comprising a pharma- ceutical composition comprising a pharma- ceutical carrier and a formulation comprising a therapeutic amount of an API, wherein the API is synthetic D-beta hydroxybutyrate;

[0418] The administration is intranasal (IN),

[0419] Administering comprises targeted delivery to the olfactory region of the nasal cavity of the subject;

[0420] Targeted delivery achieves a therapeutically effective amount of the API in the brain,

[0421] An effective amount of the pharmaceutical composition provides a method of (i) eliminating or reducing the risk of acquired brain injury, or (ii) reducing the severity of acquired brain injury, or (iii) delaying the onset of acquired brain injury, or (iv) a combination thereof.

[0422] According to some embodiments, the acquired brain injury is a traumatic brain injury. According to some embodiments, the traumatic brain injury comprises a concussion. According to some embodiments, the subject's risk of acquired brain injury is increased by participation in an event where acquired brain injury is a known risk. According to some embodiments, the event is a military operation. According to some embodiments, the event is a sporting event. According to some embodiments, the sporting event is, for example, hockey, football, soccer, baseball, polo, rugby, horse riding, auto racing, motorcycling, skiing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing, or (iv) a combination thereof. Other applications include emergency medical use, such as by EMTs or emergency departments in the context of such emergency situations as domestic violence, falls, motor vehicle accidents, and the like.

[0423] According to some embodiments, the present invention is directed to a subject within 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours or less prior to an event for which acquired brain injury is a known risk. Administration within 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, etc. is neuroprotective.

[0424] According to some embodiments, the pharmaceutical composition comprises an active agent formulated as a solution, suspension or dispersion. According to some embodiments, the API is formulated as a solution. According to some embodiments, the API is formulated as a suspension. According to some embodiments, the API is formulated as a suspension, the suspension comprising a particle size distribution of a population of particles comprising the API n. According to some embodiments, the API is formulated as a dispersion. According to some embodiments, the API is formulated as a dispersion, the dispersion comprising a particle size distribution of a population of particles comprising the API.

[0425] According to some embodiments, the pharmaceutical composition comprises a liquid spray formulation or a dry powder formulation.

[0426] According to some embodiments, the pharmaceutical composition comprises a liquid spray formulation of the API. According to some embodiments, the pharmaceutical composition is delivered as a liquid spray comprising a droplet size distribution comprising droplets containing the API.

[0427] According to some embodiments, the pharmaceutical composition comprises a dry powder formulation. According to some embodiments, the aerosolized dry powder formulation comprises a cloud of very fine particles comprising the API.

[0428] According to some embodiments, the formulation is formulated with an excipient. According to some embodiments, the excipient maximizes the solubility of the API. According to some embodiments, the excipient enhances adhesion of the API to the nasal mucosa in the olfactory system. According to some embodiments, the enhanced adhesion enhances the absorption of the API in the nasal mucosa.

[0429] According to some embodiments, the nasal aerosol comprises the formulation, the container, the valve, the actuator, the dust cap, the associated accessories, and the protective packaging, which together constitute the drug product. For administration by injection, for example, the pharmaceutical composition according to the invention described can be conveniently delivered in the form of an aerosol spray presentation from a pressurized pack using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. For pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges, for example gelatin capsules and cartridges for use in inhalers, can be formulated to contain a powder mix of the active compound and a suitable powder base, for example, lactose or starch.

[0430] According to some embodiments, the formulation is loaded into a unit dose device. According to some devices, the formulation is loaded into a multi-dose device. According to some embodiments, exemplary devices include a spray pump and a metered dose inhaler. According to some embodiments, the liquid formulation is loaded into a spray pump, which is an exemplary device for delivering intranasal formulations. According to some embodiments, the dry powder formulation is loaded into a dry powder device that includes a formulation of active agent and a container closure system.

[0431] For example, a typical nasal spray formulation consists of an active agent suspended or dissolved in an aqueous medium that is loaded into a bottle with a metered spray pump. Actuation of the pump by the subject delivers droplets containing the drug into the nasal cavity. The role of the pump, an integral part of the entire assembly, is to spray and deliver a precise drug dose.

[0432] Most nasal spray pumps produce droplets in the range of about 20 μm to about 120 μm, i.e., about 20 μm, about 25 μm, about 30 μm, about 35 μm, about 40 μm, about 45 μm, about 50 μm, 55 μm, about 60 μm, about 65 μm, about 70 μm, about 75 μm, about 80 μm, about 85 μm, about 90 μm, about 95 μm, about 100 μm, about 105 μm, about 110 μm, about 115 μm, or about 120 μm, containing a therapeutic amount of the API [Yu, G. et al. Fluid flow and particle diffusion in the human upper respiratory system. Aerosol Science and Technol. (1998) 28:146-58], such that the droplet size allows the droplets to deposit in the olfactory portion of the nasal cavity for delivery of the API to the brain. If the droplet size is too fine (less than 10 μm), the droplets may pass through the nasal passages and accumulate in the lungs. If the droplet size is too large, the spray may be trapped in the nostrils. Laser diffraction techniques can measure droplet size by measuring the intensity of light scattered by a particle as a function of angle to determine the droplet size for a given device.

[0433] The characteristics of the emitted spray plume are believed to be important in assessing pump performance. According to some embodiments, pump performance is characterized by the spray pattern and plume geometry of the plume emitted by the delivery device. During the very early life of the aqueous nasal spray plume, the formulation may exit the actuator orifice as a thin stream and then form a relatively stable, fully developed cone-shaped plume before separating from the orifice. The term "plume geometry" describes the side view of the aerosol cloud parallel to the axis of the plume. According to some embodiments, the measurement of the plume geometry includes an image taken from the side of the plume perpendicular to the laser light sheet, which is aligned with the expected central plume axis. According to some embodiments, the measurement of the plume geometry is by flash illumination of the plume with photographic image capture. According to some embodiments, the plume geometry is characterized by the spray angle and the plume width.

[0434] According to some embodiments, the pharmaceutical composition comprises a formulation containing a distribution of particles containing the API. According to some embodiments, the majority of the distribution of particles is in the range of about 25 μm to about 40 μm in diameter, i.e., about 25 μm, about 26 μm, about 27 μm, about 28 μm, about 29 μm, about 30 μm, about 31 μm, about 32 μm, about 333, about 34 μm, about 35 μm, about 36 μm, about 37 μm, about 38 μm, about 39 μm, or about 40 μm. According to some embodiments, the distribution of particles is depleted of smaller particles that would otherwise enter the lungs.

[0435] According to some embodiments, a viscosity enhancer / mucoadhesive polymer, such as microcrystalline cellulose, sodium carboxymethylcellulose, or hydroxypropylmethylcellulose, or a combination thereof, may be added to the formulation at 0.1%, 0.2%, 0.3%, 1% or 2% of the formulation by weight or volume to increase the absorption of the API at the nasal mucosa in the olfactory portion of the nasal cavity. According to some embodiments, the viscosity of each nasal spray formulation is measured. According to some embodiments, the spray pattern and plume geometry of each nasal spray formulation is measured. According to some embodiments, the droplet size distribution of each nasal spray is determined, and the median (Dv50), 10th percentile (Dv10) and 90th percentile (Dv90) of the cumulative undersize distribution are determined.

[0436] According to some embodiments, for intranasal delivery, a maximum of about 25 mg / dose may be administered every 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 hours, 74 hours, 75 hours, 76 hours, 77 hours, 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 83 hours, 84 hours, 85 hours, 86 hours, 87 hours, 88 hours, 89 hours, 90 hours, 91 hours, 92 hours, 93 hours, 94 hours, 95 hours, 96 hours, 97 hours, 98 hours, 99and the like for 8 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, etc., inclusive, ranging from about 85 mg / day (e.g., for a child weighing about 45 pounds) to about 800 mg / day (e.g., for a child weighing about 45 pounds). , for adults), i.e., at least about 85 mg / day, about 95 mg / day, about 105 mg / day, about 115 mg / day, about 125 mg / day, about 135 mg / day, about 145 mg / day, about 155 mg / day, about 165 mg / day, about 175 mg / day, about 185 mg / day, about 195 mg / day, about 205 mg / day, about 215 mg / day, about 225 mg / day, about 235 mg / day, about 245 mg / day, about 255 mg / day, about 265 mg / day, about 275 mg / day, about 285 mg / day, about 295 mg / day, about 305 mg / day, about 315 mg / day, about 325 mg / day, about 335 mg / day, about 345 mg / day, about 355 mg / day, about 365 mg / day, about 375 mg / day, about 385 mg / day, about 395 mg / day, about 405 mg / day, about 415 mg / day, about 425 mg / day, about 435 mg / day, about 445 mg / day, about 455 mg / day, about 465 mg / day, about 475 mg / day, about 485 mg / day, about 495 mg / day, about 505 mg / day, about 515 mg / day, about 525 mg / day, about 535 mg / day, about 545 mg / day, about 555 mg / day, about 565 mg / day, about 575 mg / day, about 585 mg / day, about 595 mg / day, about 605 mg / day, about 615 mg / day, about 625 mg / day, about 635 mg mg / day, about 325 mg / day, about 335 mg / day, about 345 mg / day, about 355 mg / day, about 365 mg / day, about 375 mg / day, about 385 mg / day, about 395 mg / day, about 405 mg / day, about 415 mg / day, about 425 mg / day, about 435 mg / day, about 445 mg / day, about 455 mg / day, about 465 mg / day, about 475 mg / day, about 485 mg / day, about 495 mg / day, about 505 mg / day, about 515 mg / day, about 525 mg / day, about 535 mg / day, about 545 mg / day, about 555 mg / day, about 565 mg / day, achieve a minimum daily dose of about 575 mg / day, about 585 mg / day, about 595 mg / day, about 605 mg / day, about 615 mg / day, about 625 mg / day, about 635 mg / day, about 645 mg / day, about 655 mg / day, about 665 mg / day, about 675 mg / day, about 685 mg / day, about 695 mg / day, about 705 mg / day, about 715 mg / day, about 725 mg / day, about 735 mg / day, about 745 mg / day, about 755 mg / day, about 765 mg / day, about 775 mg / day, about 785 mg / day, about 795 mg / day, or about 800 mg / day;

[0437] According to some embodiments, based on the HED from the mouse study reported in paragraph

[0634] below, the minimum daily dose for a 60 kg human (132 lbs) is 250 mg / day. According to some embodiments, the minimum daily dose for a 90 kg human (200 lbs) is 730 mg / day.

[0438] According to some embodiments, a dose-time curve is determined such that a maximum of 25 mg / dose delivered intranasally at a given frequency as tolerated, e.g., every 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes, for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, etc., achieves a desired effective amount.

[0439] According to some embodiments, the present invention is directed to a subject within 1 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours or less prior to an event for which acquired brain injury is a known risk. Intranasal administration within, within 20 hours, within 21 hours, within 22 hours, within 23 hours, within 24 hours, within 1 day, within 2 days, within 3 days, within 4 days, within 5 days, within 6 days, within 7 days, within 8 days, within 9 days, within 10 days, within 11 days, within 12 days, within 13 days, within 14 days, within 15 days, within 16 days, within 17 days, within 18 days, within 19 days, within 20 days, within 21 days, etc. is neuroprotective.

[0440] According to some embodiments, the event is a military operation. According to some embodiments, the event is a sporting event. According to some embodiments, the sporting event is, for example, hockey, football, soccer, baseball, polo, rugby, horse riding, auto racing, motorcycling, skiing, gymnastics, mountain climbing, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing. Other applications include emergency medical use, such as by EMTs or emergency departments in the context of such emergency situations as domestic violence, falls, motor vehicle accidents, and the like.

[0441] According to some embodiments, the incorporation of D-beta hydroxybutyrate into a polymeric carrier can achieve controlled release, including delayed release and sustained release.

[0442] According to some embodiments, D-beta hydroxybutyrate binds to HCA2.

[0443] According to some embodiments, the composition, when delivered intranasally, increases brain D-beta hydroxybutyrate levels for, e.g., at least 6 hours to at least 48 hours, inclusive, i.e., at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, at least 24 hours, at least The concentration may be increased to about 0.1-3 mmol / L, or to about 0.1-1 mmol / L, for at least 25 hours, at least 26 hours, at least 27 hours, at least 28 hours, at least 29 hours, at least 30 hours, at least 31 hours, at least 32 hours, at least 33 hours, at least 34 hours, at least 35 hours, at least 36 hours, at least 37 hours, at least 38 hours, at least 39 hours, at least 40 hours, at least 41 hours, at least 42 hours, at least 43 hours, at least 44 hours, at least 45 hours, at least 46 hours, at least 47 hours, or at least 48 hours.

[0444] (2) After damage

[0445] According to another aspect, the disclosure provides a method for treating symptoms of brain injury, including neuroinflammation, in a subject, comprising administering a pharmaceutical composition comprising a pharma- ceutical composition comprising a pharma- ceutical acceptable carrier and a formulation comprising a therapeutic amount of an API, wherein the API is synthetic D-beta hydroxybutyrate;

[0446] The administration is intranasal (IN),

[0447] Administering comprises targeted delivery to the olfactory region of the nasal cavity of the subject;

[0448] Targeted delivery achieves a therapeutically effective amount of the API in the brain,

[0449] An effective amount of the pharmaceutical composition comprises:

[0450] (a) reducing the severity of acquired brain injury;

[0451] (b) limiting the occurrence of symptoms characteristic of the acquired brain injury being treated;

[0452] (c) limiting the worsening of symptoms characteristic of the acquired brain injury being treated;

[0453] (d) limiting the recurrence of acquired brain injury in a subject who has previously had an acquired brain injury; or

[0454] (e) limiting the recurrence of symptoms in a subject who was previously asymptomatic for the acquired brain injury.

[0455] According to some embodiments, the acquired brain injury is a traumatic brain injury. According to some embodiments, the traumatic brain injury comprises a concussion. According to some embodiments, the subject's risk of acquired brain injury is increased by the subject's participation in an event where acquired brain injury is a known risk. According to some embodiments, the event is a military operation. According to some embodiments, the event is a sporting event. According to some embodiments, the sporting event may be, for example, hockey, football, soccer, baseball, polo, rugby, horse riding, auto racing, motorcycling, skiing, mountain climbing, gymnastics, basketball, mixed martial arts, Brazilian Jiu Jitsu, Muay Thai, Taekwondo, kickboxing, boxing, wrestling, lacrosse, softball, cheerleading, volleyball, beach volleyball, alpine skiing, water skiing, wakeboarding, snowboarding, skateboarding, kayaking, handball, cycling, mountain biking, barrel racing, bull riding, BASE jumping, skydiving, paragliding, tennis, squash, BMX, motocross, surfing, bobsleigh, luge, skeleton, broomball, hurling, camogie, cricket, diving, field hockey, figure skating, speed skating, sailing, ski jumping, ultimate frisbee, water polo, or windsurfing. Other applications include emergency medical use, such as by EMTs or emergency departments in the context of such emergency situations as domestic violence, falls, motor vehicle accidents, and the like.

[0456] According to some embodiments, administration is at a given frequency as tolerated, for example, every 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes after the event, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, within at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, or at least 21 days.

[0457] According to some embodiments, the pharmaceutical composition comprises a liquid spray formulation or a dry powder formulation. According to some embodiments, the pharmaceutical composition comprises a liquid spray formulation. According to some embodiments, the pharmaceutical composition comprises a dry powder formulation. According to some embodiments, the formulation is formulated with an excipient. According to some embodiments, the excipient maximizes the solubility of the API. According to some embodiments, the excipient enhances adhesion of the API to the nasal mucosa. According to some embodiments, the enhanced adhesion enhances absorption of the API in the nasal mucosa.

[0458] According to some embodiments, the pharmaceutical composition comprises an active agent formulated as a solution, suspension or dispersion. According to some embodiments, the API is formulated as a solution. According to some embodiments, the API is formulated as a suspension. According to some embodiments, the API is formulated as a suspension, the suspension comprising a particle size distribution of a population of particles comprising the API n. According to some embodiments, the API is formulated as a dispersion. According to some embodiments, the API is formulated as a dispersion, the dispersion comprising a particle size distribution of a population of particles comprising the API.

[0459] According to some embodiments, the pharmaceutical composition comprises a liquid spray formulation or a dry powder formulation.

[0460] According to some embodiments, the pharmaceutical composition comprises a liquid spray formulation of the API. According to some embodiments, the pharmaceutical composition is delivered as a liquid spray comprising a droplet size distribution comprising droplets containing the API.

[0461] According to some embodiments, the pharmaceutical composition comprises a dry powder formulation. According to some embodiments, the aerosolized dry powder formulation comprises a cloud of very fine particles comprising the API.

[0462] According to some embodiments, the formulation is formulated with an excipient. According to some embodiments, the excipient maximizes the solubility of the API. According to some embodiments, the excipient enhances adhesion of the API to the nasal mucosa in the olfactory system. According to some embodiments, the enhanced adhesion enhances the absorption of the API in the nasal mucosa.

[0463] According to some embodiments, the nasal aerosol comprises the formulation, the container, the valve, the actuator, the dust cap, the associated accessories, and the protective packaging, which together constitute the drug product. For administration by injection, for example, the pharmaceutical composition according to the invention described can be conveniently delivered in the form of an aerosol spray presentation from a pressurized pack using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. For pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges, for example gelatin capsules and cartridges for use in inhalers, can be formulated to contain a powder mix of the active compound and a suitable powder base, for example, lactose or starch.

[0464] According to some embodiments, the formulation is loaded into a unit dose device. According to some embodiments, the formulation is loaded into a multi-dose device. According to some embodiments, exemplary devices include a spray pump and a metered dose inhaler. According to some embodiments, the liquid formulation is loaded into a spray pump, which is an exemplary device for delivering intranasal formulations. According to some embodiments, the dry powder formulation is loaded into a dry powder device comprising a formulation of active agent and a container closure system.

[0465] For example, a typical nasal spray formulation consists of an active agent suspended or dissolved in an aqueous medium that is loaded into a bottle with a metered spray pump. Actuation of the pump by the subject delivers droplets containing the drug into the nasal cavity. The role of the pump, an integral part of the entire assembly, is to spray and deliver a precise drug dose.

[0466] Most nasal spray pumps produce droplets in the range of about 20 μm to about 120 μm, i.e., about 20 μm, about 25 μm, about 30 μm, about 35 μm, about 40 μm, about 45 μm, about 50 μm, 55 μm, about 60 μm, about 65 μm, about 70 μm, about 75 μm, about 80 μm, about 85 μm, about 90 μm, about 95 μm, about 100 μm, about 105 μm, about 110 μm, about 115 μm, or about 120 μm, containing a therapeutic amount of the API [Yu, G. et al. Fluid flow and particle diffusion in the human upper respiratory system. Aerosol Science and Technol. (1998) 28:146-58], such that the droplet size allows the droplets to deposit in the olfactory portion of the nasal cavity for deli...

Claims

1. A pharmaceutical composition for use in a subject requiring treatment for traumatic brain injury (TBI), comprising a composition comprising a therapeutic amount of D-beta hydroxybutyrate and formulated for intranasal delivery.

2. The subject has neuroinflammation, the pharmaceutical composition comprises a pharmaceutically acceptable carrier and a formulation comprising a therapeutic amount of an API, wherein the API is synthetic D-beta hydroxybutyrate; 10. The pharmaceutical composition of claim 1, wherein administration of the pharmaceutical composition is intranasal (IN), wherein the administration comprises targeted delivery to the olfactory region of the nasal cavity of the subject, wherein the targeted delivery achieves a therapeutic amount of the API in the brain.

3. The pharmaceutical composition according to claim 1, wherein an effective amount of the pharmaceutical composition is: (a) reducing the severity of acquired brain injury; (b) limiting the occurrence of symptoms characteristic of the acquired brain injury being treated; (c) limiting the worsening of symptoms characteristic of the acquired brain injury being treated; and (d) limiting the recurrence of the acquired brain injury in a subject who has had a history of the acquired brain injury, or the recurrence of symptoms in a subject who was previously asymptomatic for the acquired brain injury; The pharmaceutical composition of claim 2, which achieves one or more of the above.

4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is administered to a subject after the subject has experienced an external force to the head, and optionally within 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, or 24 hours after the subject has experienced the external force.

5. The pharmaceutical composition of claim 1, wherein the D-beta hydroxybutyrate is a salt or ester of D-beta hydroxybutyrate.

6. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is formulated as a spray or a dry powder.

7. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is administered multiple times.

8. A pharmaceutical composition comprising a therapeutic amount of D-beta hydroxybutyrate and formulated for intranasal delivery for use in subjects in need of prophylactically limiting the development of symptoms characteristic of traumatic brain injury (TBI).

9. A pharmaceutical composition as described in claim 8 for use in a method for reducing the risk of brain damage resulting from acquired brain injury in a subject who is susceptible to or at high risk of said acquired brain injury.

10. The pharmaceutical composition comprising a pharmaceutically acceptable carrier and a formulation comprising a therapeutic amount of an API, wherein the API is synthetic D-beta hydroxybutyrate; 10. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is administered intranasally (IN), wherein the administration comprises targeted delivery to the olfactory region of the nasal cavity of the subject, wherein the targeted delivery achieves a therapeutically effective amount of the API in the brain.

11. An effective amount of the pharmaceutical composition comprising: (i) eliminating or reducing the risk of said acquired brain injury; (ii) reducing the severity of said acquired brain injury; (iii) delaying the onset of said acquired brain injury; or (iv) combinations thereof The pharmaceutical composition according to claim 10,

12. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is administered before the subject engages in an activity that increases the subject's risk of suffering a traumatic brain injury (TBI), and optionally within 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, or 24 hours of the activity.

13. The pharmaceutical composition of claim 8, wherein the D-beta hydroxybutyrate is a salt or ester of D-beta hydroxybutyrate.

14. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is formulated as a spray or dry powder.

15. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is administered multiple times.

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