ALPHA-1062 for Treating Traumatic Brain Injury

Abstract

The present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, or the compound ALPHA-1062 or a salt thereof, for use in treating a confirmed or suspected traumatic brain injury (TBI) in a subject. In some embodiments, the composition is administered transmucosally, for example intranasally, and / or the composition is self-preserving and antimicrobial. The present invention also relates to a multi-use dispenser configured for intranasal or transmucosal administration of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof. The present invention further relates to a method of treating traumatic brain injury (TBI) in a subject, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof.

Description

[Technical field]

[0001] The present invention is in the field of medical methods as well as compositions, formulations and dispensing devices for pharmaceutical agents.

[0002] The present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, or the compound ALPHA-1062 or a salt thereof, for use in treating a confirmed or suspected traumatic brain injury (TBI) in a subject. In some embodiments, the composition is administered transmucosally, for example intranasally, and / or the composition is self-preserving and antimicrobial. The present invention also relates to a multi-use dispenser configured for intranasal or transmucosal administration of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof. The present invention further relates to a method of treating traumatic brain injury (TBI) in a subject, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof. [Background technology]

[0003] Traumatic brain injury (TBI) is a global public health problem and a leading cause of death and disability. TBI is typically classified clinically as mild, moderate, or severe at the time of injury using the Glasgow Coma Scale (GCS). Severe TBI can result in mortality rates as high as 30-40%. Survivors experience a substantial burden of physical, psychiatric, emotional, and cognitive impairments that disrupt the lives of individuals and their families. Such impairments are not limited to severe cases, but also frequently occur after moderate or mild TBI.

[0004] The effects and outcomes of TBI are influenced not only by the severity of the injury, but also by factors such as repetitive events and age. TBI can result from a variety of injury mechanisms, including falls, motor vehicle or other types of traffic injuries, sports injuries, and interpersonal physical or other violence (e.g., blast injuries). With respect to age, TBI shows a bimodal distribution with the highest incidence in the youngest and oldest age groups. These age groups are more likely to suffer TBI due to increased risk of various injury causes, such as falls in children under 4 years and elderly over 75 years, and motor vehicle accidents in adults.

[0005] Sports-related TBI is increasingly being described as a chronic health condition, with evidence suggesting that the health effects of TBI may persist over time, especially in those with moderate to severe TBI. While the majority of patients with mild TBI (mTBI) recover quickly, the effects of mTBI may persist for more than a year. Symptoms often associated with long-term effects include cognitive impairments such as attention deficits, memory problems, and executive dysfunction or even social deficits. The long-term effects of mTBI may range from impaired social reasoning (interpretation of verbal and non-verbal social cues) to psychological symptoms such as anxiety, or physical symptoms such as fatigue, balance and coordination problems. mTBI may be associated with persistent headaches, vestibular dysfunction, depression, and cognitive complaints. The cumulative effects may result in symptoms such as cognitive impairment, behavioral disorders, mood disorders, and movement disorders.

[0006] Repeated injuries have been most thoroughly studied in contact sports such as American football, where athletes have an increased risk of death by suicide, reduced cognitive function, macrostructural, microstructural, functional and neurochemical changes, and an increased risk of death from neurodegenerative causes such as dementia or Alzheimer's disease. Another cause of TBI could be blast-related TBI in military personnel (Haarbauer-Krupa et al., J Neurotrauma, 2021; Brazinova et al., J Neurotrauma, 2021; Center-TBI project; American Association of Neurological Surgions (AANS)).

[0007] Today, no curative treatment is available for TBI itself. Current drug therapies aim to reduce secondary brain injury that occurs as a downstream effect or symptom of TBI. Such drugs include anti-seizure medications for patients experiencing seizures after moderate or severe TBI, coma-inducing drugs that aim to reduce oxygen consumption in the injured brain, and diuretics that can reduce pressure within the brain. The only other available treatment options for TBI are surgical intervention, physical and mental rest, or rehabilitation therapies that target the long-term effects of TBI.

[0008] Galantamine and its derivatives have been suggested for the treatment of neurodegenerative diseases such as Alzheimer's disease and dementia. Unfortunately, like other cholinesterase inhibitors, galantamine has clinically significant levels of mechanism-based gastro-intestinal (GI) side effects, including nausea, vomiting, and diarrhea (Loy C et al., Galantamine for Alzheimer's disease and mild cognitive impairment. Cochrane Database of Systematic Reviews 2006). To accommodate patients to these side effects, cholinesterase inhibitors are usually administered at low (non-effective) doses initially, and the dose is carefully titrated to an effective dose within 4-6 weeks. Furthermore, the maintenance dose is often adjusted to what the patient experiences as a tolerable level of GI side effects, and it is likely that most, if not all, patients will never achieve treatment with the most effective dose. Thus, cholinesterase inhibitors such as galantamine are not considered optimal for addressing TBI, where an immediately effective dose is preferred.

[0009] To enhance the lipophilicity of acetylcholinesterase inhibitors and improve their passage through mucosal tissues, hydrophobic side chains have been added to the basic alkaloid structure.Galanthamine derivatives and prodrugs are described in EP 1940817, WO 2009 / 127218, and U.S. Patent Application Publication No. 2009 / 0253654.

[0010] The galantamine prodrug ALPHA-1062 is a benzoic acid or benzoate ester of galantamine ((4aS,6R,8aS)-5,6,9,10,11,12-hexahydro-3-methoxy-11-methyl-4aH-[1]benzofuro[3a,3,2-ef][2]benzazepin-6-ol benzoate). It was developed to enhance the hydrophobicity of galantamine. ALPHA-1062 is essentially pharmacologically inactive until cleaved by esterases resulting in the release of galantamine.

[0011] WO 2014 / 016430 discloses various formulations and salts of ALPHA-1062, including, for example, lactate, gluconate, maleate, and saccharate salts, as well as transmucosal administration of ALPHA-1062 via intranasal, buccal, or sublingual modes.

[0012] The first nasal spray pump for intranasal administration of liquid compositions was developed approximately 50 years ago, replacing earlier step-by-step droppers and pipettes. Nasal spray pumps are now widely used to moisten the nasal mucosa with saline, as nasal preparations for the administration of locally acting drugs, such as nasal decongestants, or for the non-invasive administration of substances that need to reach the systemic circulation, such as anti-migraine drugs or hormones (Marx and Birkhoff, "Multi-Dose Container for Nasal and Ophthalmic Drugs: A Preservative Free Future?" in Drug Development-A Case Study Based Insight into Modern Strategies, ed. Chris Rundfeldt, 2011).

[0013] For many disease indications, multi-dose devices are a cost-effective and convenient means to provide the safety and accuracy of administration of active agents required by regulatory agencies. However, to date, most drugs administered transmucosally, generally as solutions, emulsions or suspensions, contain preservatives to support long-term storage times and stability during proper use of multi-use or multi-dose dispensers. However, despite the benefits of nasal administration, the use of preservatives in nasal sprays is controversial. Reports suggest that preservatives in nasal sprays may increase the risk of adverse events for patients.

[0014] Tenovuo et al (Progress in Neuro-Psychopharmacology & Biological Psychiatry,vol.29,no.1,p.61-67,2005) teach the oral administration of donepezil, galantamine, or rivastigmine as treatment for chronic stable TBI. The study was not placebo controlled and did not perform clinical trials after drug administration to verify the reported symptomatic improvement. Patients with chronic TBI reported subjective improvement in TBI-related symptoms after treatment, but approximately half of all patients experienced worrisome adverse effects. Despite studies conducted almost 20 years ago, none of these drugs have been adopted as standard of care in the management of either acute or chronic TBI.

[0015] Thus, there is a lack of first-line treatments for TBI, particularly in acute settings, that can reduce or prevent secondary downstream damage and short- and / or long-term symptoms of TBI, such as cognitive and neurological impairments. There is an urgent need to provide effective treatments for TBI. Summary of the Invention

[0016] In light of the prior art, the technical problem underlying the present invention was the provision of an improved or alternative means for treating traumatic brain injury (TBI). Another problem underlying the present invention was to provide a convenient, multi-use, preferably preservative-free formulation suitable for administering an effective amount of an active pharmaceutical agent in treating TBI.

[0017] This problem is solved by the features of the independent claims. Preferred embodiments of the invention are provided by the dependent claims.

[0018] Accordingly, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in the treatment of confirmed or suspected traumatic brain injury (TBI) in a subject.

[0019] In other embodiments or aspects, the present invention relates to the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject.

[0020] In other embodiments or aspects, the present invention relates to a method of treating traumatic brain injury (TBI) in a subject, comprising administering a pharma- ceutically effective amount of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof.

[0021] The present invention is based on the finding that ALPHA-1062 shows efficacy in the treatment of TBI.As will be discussed in more detail below, an exemplary 35-day study using a controlled cortical impact injury model was carried out, indicating the efficacy of ALPHA-1062.The positive therapeutic effect on TBI may include, but is not limited to, improved functional and histological outcomes after traumatic brain injury (TBI).

[0022] One difference between the previous published study and the current ALPHA-1062 study is the degree of acute protection achieved through ALPHA-1062 administration, which appears to reduce the degree of functional impairment on day 1 after TBI, as well as the rate and extent of sustained improved recovery, all compared to vehicle treatment.Compared to published literature, ALPHA-1062 treatment had a stronger effect than galantamine in terms of both functional recovery and neuropathological reduction.Those skilled in the art would not have expected that ALPHA-1062 would enable these advantages over galantamine administration.

[0023] The inventors surprisingly observed that ALPHA-1062 administration preserved brain structures, reduced neuronal cell loss, and promoted neurogenesis after TBI. The inventors found that ALPHA-1062 treatment after TBI (compared to vehicle control treatment) preserved brain structures, such as hippocampal structures, and significantly enhanced neurogenesis, including an increase in neuroblasts (verified by tissue staining for neurogenesis markers DCX or BrdU / NeuN+). ALPHA-1062 treatment also significantly reduced lesion size measured 35 days after TBI injury and significantly reduced neuronal cell loss in brain regions affected by TBI, such as the cortical and hippocampal regions of the injured brain. Unexpectedly, the degree of neuroprotection was so strong that the neuronal cell counts determined in these regions of animals subjected to TBI and treated with ALPHA-1062 were indistinguishable from the neuronal cell counts determined for sham-treated [non-injured] animals.

[0024] Moreover, ALPHA-1062 treatment significantly reduced the accumulation of pathological phosphorylated forms of Tau (AT-8 positive), known to accumulate in the brains of TBI and Alzheimer's patients. Without wishing to be bound by theory, the reduction in p-Tau accumulation observed after ALPHA-1062 treatment of TBI in a rat study disclosed in detail below may reduce the risk of later developing dementia in ALPHA-1062-treated TBI patients. In some cases, even a single mild TBI without syncope can significantly increase the risk of dementia by about two-fold, as shown in a cohort of US military personnel evaluated longitudinally (Barnes et al, Association of mild traumatic brain injury with and without loss of consciousness with dementia in US military veterans. JAMA neurology, 2018, 75(9), 1055-1061). In Alzheimer's disease, pathological Tau accumulation precedes beta-amyloid accumulation by several years. Pathologically phosphorylated Tau is believed to spread by "seeding" in a similar manner to prion protein, with a relatively slow development with a predicted doubling time of approximately 5 years. Considering that TBI can induce (or be associated with) an increased long-term risk of dementia, it is plausible to assume that ALPHA-1062 treatment, which has been shown to reduce pathologically phosphorylated Tau, may also show the benefit of reducing the risk of later-onset neurological defects. The observation of reduced p-Tau accumulation in the TBI model disclosed below is entirely novel, and this technical effect has not been shown or suggested before for galantamine.

[0025] In summary, we found that ALPHA-1062 treatment is effective against moderate TBI and significantly improves recovery of brain tissue, sensorimotor, and cognitive function abilities when administered acutely after injury. To the best of our knowledge, the observed improvements could not be predicted from the prior art for the TBI experimental group.

[0026] In one embodiment, the composition is in liquid form.

[0027] In embodiments, the liquid is a solution, emulsion, or suspension.

[0028] In another embodiment, the composition is in a solid form.

[0029] In embodiments, the solid form is a tablet, pill, film, lozenge, or capsule.

[0030] The present invention also relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the composition is administered transmucosally.

[0031] In some embodiments, the composition is administered intranasally.

[0032] In embodiments, the composition is administered buccally.

[0033] In embodiments, the compositions are administered sublingually or bucally.

[0034] Advantageous transport properties can be achieved for prodrugs when administered by intravenous injection, but less well or to a lesser extent when administered orally, e.g., as tablets for systemic administration, because prodrug esters are unstable in acidic environments (such as those present in the stomach) and can be enzymatically cleaved in many tissues, including the intestine and liver (first-pass effect).

[0035] In light of these findings and the problems with prior art administration methods, and to utilize ALPHA-1062 in the treatment of CNS disorders, in some embodiments, the present invention utilizes an administration route that avoids the gastrointestinal tract and the first-pass effect (loss of a certain percentage of drug due to metabolism in the intestine and liver). These administration routes of ALPHA-1062 provide brain delivery of the active drug galantamine at levels similar to intravenous injection of ALPHA-1062. Thus, the present invention employs, in some embodiments, pharmaceutical formulations used for selected administration routes that optimize rapid absorption and uptake of ALPHA-1062.

[0036] Of note, mucosal administration of galantamine as in the prior art does not provide such enhancement, since galantamine is not susceptible to cleavage by endogenous esterases. The surprising concept of mucosal administration of ALPHA-1062 in treating TBI is based on avoiding cleavage of the prodrug after administration but before distribution through the BBB, thereby enhancing brain transport and increasing the relative concentration of the active substance after cleavage, which occurs significantly in the brain under the conditions of the proposed administration route and drug formulation.

[0037] Thus, the present invention provides improved means and formulations that can be administered shortly after the occurrence of brain trauma in an effective and easy-to-use form that exhibits low side effects and allows for good patient compliance.

[0038] As shown in the examples, the inventors have surprisingly developed application and treatment methods that reduce the short-term and long-term effects of TBI, such as cognitive, neurological, and motor dysfunction. The formulations and compounds according to the present invention show surprisingly beneficial effects in the treatment of subjects with TBI or potentially other brain trauma. Treatment of subjects with TBI events with formulations and compounds according to the present invention, including ALPHA-1062 or a salt thereof, results in better recovery of motor skills, as well as spatial and recognition memory, compared to vehicle treatment. Treatment of subjects with TBI with ALPHA-1062 or a salt thereof according to the present invention appears to cause excellent recovery of motor and memory skills, as well as other cognitive functions, which are comparable to those in healthy individuals without TBI. This surprising effect is complemented by the transmucosal administration of the compounds and compositions according to the present invention, which not only facilitates improved patient compliance due to its easy administration and reduced adverse effects, but also achieves good efficacy of the compounds due to direct delivery of the compounds to the brain (avoiding the first-pass effect). This surprising effect may represent a significant improvement over current treatment options for TBI and is demonstrated by the experimental results presented in the Examples.

[0039] Thus, the present invention provides a novel and surprising solution for treating, reducing, and / or preventing (or reducing the risk of) TBI-related short-term and long-term symptoms, such as acute and / or chronic neurological and cognitive impairments.

[0040] Thus, the present invention in one embodiment relates to ALPHA-1062 for use as a medicament in the treatment of TBI as described herein, wherein the transmucosal administration is configured to avoid and / or reduce immediate cleavage of the ester group of the agent by endogenous esterases, e.g. during and / or after absorption from the gastrointestinal tract.

[0041] This aspect of the present invention, namely, addressing the relatively poor stability of the ester moiety of ALPHA-1062 in the gastrointestinal tract and liver, represents a novel technical effect in the treatment of TBI not previously disclosed or suggested in the art.

[0042] With regard to the improvements afforded by the transmucosal administration and enhanced delivery of the compounds and salts described herein, avoidance of in vivo esterase cleavage allows for the treatment of patients who previously avoided or discontinued treatment with AChE inhibitors due to the strong gastrointestinal side effects associated with oral administration of effective doses of galantamine, without the typical 4-6 week titration period required for tolerability. Improved brain delivery by transmucosal administration, particularly with highly concentrated aqueous solutions of ALPHA-1062 salts administered intranasally, allows for dosing regimens that were previously difficult or impractical for galantamine (due to significant side effects) or ALPHA-1062 (due to in vivo degradation by the oral route).

[0043] The present invention also relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the composition is self-preserving and antimicrobial, preferably the composition is essentially free of added antimicrobial preservatives.

[0044] As described in detail below, the present invention is based on the surprising discovery of the beneficial effect of the use of ALPHA-1062 in the treatment of TBI. The indication is not clear in the prior art where this drug (ALPHA-1062) has antibacterial properties. As described in the introduction, ALPHA-1062 was known to show effects on cognitive impairment by acting as a prodrug of galantamine. There is no suggestion in the art that galantamine or its prodrug ALPHA-1062 may show properties to actively reduce the CFU / mL of pathogenic bacteria, yeasts, and fungi as demonstrated according to the USP 51 test.

[0045] The identification of this novel property (antibacterial effect) of a known substance (ALPHA-1062) opens new clinical perspectives and creates new clinical situations when considering either the formulation or administration of the substance. With the discovery of a novel property, a new patient population is served and, based on the surprising property, new dosing regimens and formulation options can now be adopted.

[0046] Providing the self-preserving ALPHA-1062 composition, preferably as a liquid, or more preferably as an emulsion or solution, for example in the form of a multi-use dispenser without added preservatives, allows for a reduction in side effects, for example those caused by nasal administration of a solution containing added preservatives. The storage properties of any given formulation may now be improved, i.e., the discovery of antibacterial properties may reveal longer storage times.

[0047] In some embodiments, a reduction in side effects may be achieved, which may be associated with nasal irritation (i.e., caused by the preservative), including, but not limited to, increased mucosal swelling and nasal hypersensitivity, type IV hypersensitivity, reduced mucociliary clearance, and nasal mucosal dysplasia.

[0048] In some embodiments, the compositions of the present invention do not contain one or more additional preservatives, preferably no additional preservatives, selected from the list consisting of benzalkonium (preferably benzalkonium chloride), benzyl alcohol, thimerosal (merthiolate), edetate disodium, monobasic sodium phosphate, providone, dibasic sodium phosphate, edetate disodium, monobasic potassium phosphate, providone, dibasic sodium phosphate, disodium eta, monobasic potassium phosphate, iodine, phenylcarbinol, and sodium silicoaluminate.

[0049] Furthermore, providing a self-preserved ALPHA-1062 solution, such as in the form of a multi-use dispenser without added preservatives, allows for better patient compliance compared to single-use or single-dose, as described in the art. Providing a simple spray dispenser that allows multiple uses but does not have preservatives is a simple and less side-effect-prone option for patients. First, lower mucous membrane or nasal irritation will enhance patient compliance, for example, as discomfort in the nose and / or mouth is reduced. Second, by using an easy-to-use dispenser and retaining this single dispenser for multiple uses over time, administration becomes easier for cognitively and / or neurologically impaired and / or elderly subjects, or when the treatment is administered to infants or children.

[0050] In contrast, daily administration of tablets or single dose dispensers intended for swallowing is associated with additional complications, potentially requiring patients who are children or cognitively and / or neurologically impaired to either swallow tablets or continually open, apply and purchase single dose dispensers, resulting in complications and delivery burdens. As a result, the dosage forms described herein of the compositions and compounds according to the invention ensure or at least support regular and / or long-term administration of the compounds according to the invention, thereby reducing adverse effects in the treatment of TBI and improving patient compliance, further improving the treatment outcome of TBI.

[0051] Thus, the multi-use dispenser of the present invention may be kept in a first aid kit or in the first response equipment of emergency medical personnel. The antimicrobial characteristics of ALPHA-1062, combined with its effective effectiveness in treating TBI and / or its symptoms, allows for a multi-use dispenser that does not require frequent refills or replacement, making ALPHA-1062 an ideal active agent in, for example, a pre-prepared emergency medical kit.

[0052] In one embodiment, a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof for use in treating a confirmed or suspected traumatic brain injury (TBI) in a subject comprises an amount of ALPHA-1062 sufficient to have an antimicrobial effect. Such antimicrobial effect, in some embodiments, refers to an antimicrobial effect in a pharmaceutical formulation, or an in vivo effect after administration, such as in maintaining a relatively microbial-free or microbial-poor environment within the formulation, which may result in a reduction in the microbial load in the area of ​​administration.

[0053] In one embodiment, a pharmaceutical composition comprising ALPHA-1062 or a salt thereof for use in the treatment of confirmed or suspected traumatic brain injury (TBI) in a subject is characterized in that the compound ALPHA-1062 or a salt thereof is present in a concentration of 1 to 200 mg / mL, preferably 5 to 100 mg / mL.

[0054] In some embodiments, ALPHA-1062 or a salt thereof is present in the composition at a concentration of 1 to 500 mg / mL, preferably at a concentration of 1 to 400 mg / mL, more preferably at a concentration of 1 to 300 mg / mL, preferably at a concentration of 1 to 200 mg / mL, preferably at a concentration of 5 to 100 mg / mL.

[0055] In some embodiments, ALPHA-1062 or a salt thereof is present at about 5 mg / mL, or about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 75 mg / mL, about 80 mg / mL, about 85 mg / mL, about 90 mg / mL, about 95 mg / mL, about 100 mg / mL, about 110 mg / mL, about 120 mg / mL, about 130 mg / mL, about 140 mg / mL, or about 150 mg / mL. Ranges consisting of any given value form the preceding values ​​are also contemplated.

[0056] In some embodiments, ALPHA-1062 is present in the composition as a salt, preferably lactate, gluconate, maleate, or saccharate salt. The production of ALPHA-1062 salts is described in the art, for example in WO 2014 / 016430, and can be carried out without undue burden.

[0057] In some embodiments, the salts include stoichiometric and / or non-stoichiometric salts and / or hydrates of chemical entities according to ALPHA-1062, whereby the salts are preferably described as follows: ALPHA-1062·n HX·m H 2 O. In the formula, n and m=0 to 5, n and m may be the same or different, and HX is an acid, preferably selected from lactic acid, gluconic acid, maleic acid, or saccharinic acid.

[0058] In some embodiments, other acids may be employed in the ALPHA-1062 salt formation.

[0059] Acids useful for preparing the pharma- ceutically acceptable acid addition salts according to the invention include inorganic and organic acids, such as sulfamic acid, amidosulfonic acid, 1,2-ethanedisulfonic acid, 2-ethylsuccinic acid, 2-hydroxyethanesulfonic acid, 3-hydroxynaphthoic acid, acetic acid, benzoic acid, benzenesulfonic acid, carboxylic acid, ethylenediaminetetraacetic acid, camphorsulfonic acid, citric acid, dodecylsulfonic acid, ethanesulfonic acid, ethenesulfonic acid, ethylenediaminetetraacetic acid, fumaric acid, glubionic acid, glucoheptonic acid, gluconic acid, glutamic acid, hexylresorcinic acid, hydrobromic acid, hydrochloric acid, isethionocic acid, and the like. oc) acid, (di)carbonic acid, tartaric acid, hydroiodic acid, lactic acid, lactobionic acid, laevulinic acid, lauryl sulfuric acid, lipoic acid, malic acid, maleic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalenesulfonic acid, nitric acid, oxalic acid, pamoic acid, pantothenic acid, perchloric acid, phosphoric acid, polygalacturonic acid, pectinic acid, propionic acid, salicylic acid, succinic acid or sulfuric acid, p-tuluenesulfonic acid (herein hydrochloric, hydrobromic, sulfuric, nitric, phosphoric and perchloric acids as well as tartaric, citric, acetic, succinic, maleic, fumaric and oxalic acids).

[0060] It was surprising that pharma- ceutically applicable solution embodiments of ALPHA-1062 salts met the appropriate stability, concentration, pH, osmolality, and nasal mucosal tolerability criteria in solution for intranasal application in the treatment of TBI.

[0061] In one embodiment of the present invention, the pharmaceutical composition comprises a crystalline solid form of ALPHA-1062 gluconate.In one embodiment, the crystalline solid form of ALPHA-1062 gluconate is Form A, as disclosed in WO2022 / 150917.

[0062] Polymorph studies were performed with ALPHA-1062 utilizing various solvents and crystallization conditions (Table 5 of WO 2022 / 150917) and subsequent XRPD analysis as described therein. Seven unique crystalline materials were observed and isolated, designated Form A, Form B, Form C, Form D, and Material E, Material F, and Material G (Figure 6 of WO 2022 / 150917). Amorphous material was also observed. Form A of ALPHA-1062 is an anhydrous crystalline material that begins to melt / decompose simultaneously around 117°C. Form A appears to be kinetically stable in the solid state at 43% RH (RT) and has been held at these conditions for up to 5 days. Based on WO 2022 / 150917, anhydrous Form A, stored under appropriate temperature and humidity conditions to maintain form and stability, is believed to be most suitable for the various ALPHA-1062 forms used in formulations and used in the manufacture of drug products.

[0063] In one embodiment, the present invention relates to a crystalline solid form of ALPHA-1062 gluconate (Form A), which has prominent peaks at 3.61, 10.98, 14.41, and 18.44 degrees 2-theta (±0.2) in an X-ray powder diffraction pattern.

[0064] In one embodiment, Form A has one or more additional prominent peaks at 15.20, 17.31, 17.79, 22.77, 23.64, 24.88, and 34.31 degrees 2-theta (±0.2) in its powder X-ray diffraction pattern, which are selected from the prominent peak list provided in WO 2022 / 150917 and do not appear to show substantial overlap with the prominent peaks in the XRPD patterns of Forms B through D or Materials E through G. In one embodiment, Form A has at least five prominent peaks selected from the list consisting of 3.61, 10.98, 13.80, 14.41, 14.56, 15.08, 15.20, 17.02, 17.31, 17.79, 18.44, 19.24, 20.18, 20.91, 21.22, and 22.40 degrees two-theta (±0.2) in an X-ray powder diffraction pattern.

[0065] The use of ALPHA-1062 in Form A represents a preferred embodiment of the present invention, since the polymorphic stability of Form A can be maintained during storage and after formulation. Form A can also enable high drug solubility and effective therapeutic effect. Thus, Form A can be used in the preparation of a solution to be administered to a subject, or directly in a solid dosage form.

[0066] In one embodiment, the ALPHA-1062 salt has a solubility in water of at least 10%, preferably greater than 20%, or more preferably greater than 30% (w / v). High solubility allows higher concentrations of the compound to be administered in smaller volumes, thereby further enhancing administration, for example, via mucosal administration.

[0067] A preferred embodiment of transmucosal administration represents a beneficial delivery mode due to a combination of factors. The enhanced solubility of ALPHA-1062 salt allows for higher concentrations of ALPHA-1062 to be administered, thereby allowing for greater activation of the active substance (galantamine) in the brain after cleavage. In some embodiments, the prodrug properties of ALPHA-1062 are exploited and enhanced by the transmucosal application of ALPHA-1062 salt.

[0068] Accordingly, the present invention also relates to a method of treating confirmed or suspected brain damage associated with cognitive and / or neurological disorders in a subject, comprising administering a therapeutically effective amount of ALPHA-1062 or a salt thereof to a subject in need thereof.

[0069] In one embodiment, the pharmaceutical composition of the present invention has the compound ALPHA-1062 present as the gluconate salt, preferably at a concentration of 50-100 mg / mL, more preferably at a concentration of 70-90 mg / mL, or alternatively at a concentration disclosed herein. Those skilled in the art will recognize that the concentration of active agents will vary according to common practice.

[0070] In one embodiment, the pharmaceutical composition of the present invention provides a method for the prevention and / or treatment of 1×10 Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and / or Aspergillus brasiliensis within 14 days of treatment according to United States Pharmacopeia Chapter 51 Preservation Testing (USP 51). 4 ~1×10 6 ALPHA-1062 is characterized by being present in a concentration sufficient to reduce colony forming units / mL (CFU / mL) to less than 100 CFU / mL, preferably less than 10 CFU / mL. ALPHA-1062 exhibits antibacterial activity against both gram-positive and gram-negative bacteria, pathogenic yeasts and fungi, thereby indicating a broad-spectrum antibacterial effect. There is no suggestion in the prior art that ALPHA-1062 can exhibit such broad and effective antibacterial activity and that the treatment of TBI could benefit therefrom, and thus exploiting this property of ALPHA-1062 in the treatment of TBI thus represents a surprising and unexpected discovery with practical utility.

[0071] In some embodiments, the liquid composition is stable, i.e., exhibits sufficient drug or prodrug stability and low microbial load, for an extended period of time. In some embodiments, the extended period of time is about 1 week, or 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, or more. In some embodiments, the composition is stable for about 1 month, or 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months. During this period or during storage, the composition can be administered via the multi-use device of the present invention. In some cases, storage of the composition can be for about 1 year, about 2 years, about 3 years, about 4 years, or about 5 years (preferably without use by the subject).

[0072] In some embodiments, the composition is present in a multi-use dispenser configured for transmucosal administration.

[0073] In some embodiments, the composition is present in a single-use dispenser configured for transmucosal administration.

[0074] In embodiments, the composition is present in a suitable metered dose device, such as an atomizer, spray, pump spray, dropper, squeeze tube, squeeze bottle, pipette, ampoule, nasal cannula, metered dose device, nasal spray inhaler, nasal continuous positive air pressure device, and / or breath actuated bi-directional delivery device.

[0075] In one embodiment, a therapeutically effective amount of the compound is administered using a suitable metered dose multi-use administration device or dispenser, such as a multi-use atomizer, multi-use spray, multi-use pump spray, multi-use dropper, multi-use squeeze tube, or bottle multi-use metered dose device, or a multi-use nasal spray or inhaler.

[0076] In one embodiment, the therapeutically effective amount of the compound is administered using a suitable single-use administration device or dispenser, hi several embodiments, the single-use administration device or dispenser is selected from the group including a single-use dropper, a single-use squeeze tube or bottle, and a single-dose powder dispenser.

[0077] In one embodiment, a therapeutically effective amount of ALPHA-1062, or a salt thereof, is administered to the oral cavity.

[0078] In one embodiment, a therapeutically effective amount of the compound ALPHA-1062 or a salt thereof is administered under the tongue (sublingually) by dispensing a quantity of the compound, preferably in the form of a solution or emulsion, from a multi-use dispenser and / or by spraying a preselected volume of a liquid composition, preferably a solution or emulsion, from a multi-use dispenser containing the compound or a salt thereof under the tongue.

[0079] In one embodiment, a therapeutically effective amount of the compound or salt thereof is administered to the buccal vestibule in the mouth, between the cheek and gums, preferably as a solution or emulsion, from a multi-use dispenser.

[0080] In a further aspect of the invention, in the non-limiting context of a multi-use dispenser, the invention combines the properties, features, and advantages of a multi-use transmucosal delivery device with the antimicrobial properties of ALPHA-1062 or a salt thereof. Thus, a multi-use dispenser according to the invention configured for transmucosal administration of a pharmaceutical composition in liquid form contains a self-preserving antimicrobial solution having ALPHA-1062 or a salt thereof as described herein.

[0081] In one embodiment, administration to the oral cavity is carried out by placing one or more drops of the solution or emulsion into the oral cavity, by spraying a preselected volume of a liquid composition into the oral cavity, or by administering a sublingual tablet, film formulation, lozenge, orally disintegrating tablet, or orally dispersible tablet into the oral cavity.

[0082] In one embodiment, a therapeutically effective amount of the compound is administered to the buccal vestibule in the mouth, between the cheek and gums, preferably as a solution or emulsion, from a multi-use dispenser.

[0083] In one embodiment, the multi-use dispenser is configured for intranasal administration.

[0084] In one embodiment, the multi-use dispenser is configured to dispense into the oral cavity.

[0085] In one embodiment, the multi-use dispenser is configured for transmucosal administration.

[0086] In one embodiment, the multi-use dispenser described herein is for use in the treatment of confirmed or suspected traumatic brain injury (TBI) in a subject (or for use in a corresponding method of treatment), the treatment comprising administering multiple doses of a pharmaceutical composition, which is preferably a self-preserving antimicrobial agent as described herein, to the subject from the same dispenser.

[0087] Thus, in another aspect, the present invention relates to a multi-use dispenser configured for intranasal administration of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof in liquid form for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject.

[0088] Thus, in another aspect, the present invention also relates to a multi-use dispenser configured for administration to the oral cavity of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof in liquid form for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject.

[0089] In a further aspect, the present invention relates to a single-use dispenser configured for intranasal administration of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof in liquid form for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject.

[0090] In another aspect, the present invention also relates to a single-use dispenser configured for administration to the oral cavity of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof in liquid form for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject.

[0091] The following embodiments relate to the administration of compositions comprising the compound ALPHA-1062 or a salt thereof disclosed herein, as well as the pharmaceutical compositions, both single-use and multi-use dispensers disclosed herein.

[0092] In one embodiment of a multi-use dispenser configured for intranasal administration of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof in liquid form for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, the multi-use dispenser is a suitable metered dose multi-use administration device or dispenser, such as an inhaler, an atomizer, a spray, a pump spray, a dropper, a squeeze tube, a squeeze bottle, a pipette, an ampoule, a nasal cannula, a metered dose device, a nasal spray inhaler, a nasal continuous positive air pressure device, an exhalation actuated bidirectional delivery device, a multi-use atomizer, a multi-use spray, a multi-use pump spray, a multi-use dropper, a multi-use squeeze tube or bottle, a multi-use metered dose device, and / or a multi-use nasal spray.

[0093] In one embodiment of a multi-use dispenser configured for intranasal or transmucosal administration of a pharmaceutical composition comprising ALPHA-1062 or a salt thereof in liquid form for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, the multi-use dispenser dispenses a single-dosing event volume of between 10 μL and 200 μL.

[0094] In one embodiment of the multi-use dispenser, the dispenser further includes different actuators and / or neck finishes known in the art, such as screw-on, snap-on, and crimp-on.

[0095] Preferred dispensers of the present invention relate to any of the multi-use devices described above. In some embodiments, dispensers available from, for example, Nemera (La Verpilliere, France) or Aptar Pharma (Illinois, USA) are preferred.

[0096] Nasal sprays may be contaminated by bacterial contamination from the external environment or patient, or from the air, through their drug delivery orifices. Thus, in some embodiments, nasal sprays may be employed in which there is a filter system that stops bacteria from entering the container to prevent contamination by air entering the device. Since airborne bacteria are typically around 0.3 μm, an appropriate size filter may be selected. Furthermore, recent studies have demonstrated that bacterial migration through filter membranes occurs during filtration operations, even when the pore size is significantly smaller than the size of the bacteria. Thus, in some embodiments, devices are employed that utilize silicone membranes to filter the return air.

[0097] In some embodiments, the multi-use dispenser employs a membrane (preferably made of silicone) that prevents bacteria from entering the reservoir or pump device. In one embodiment of the multi-use dispenser according to the present invention, the multi-use dispenser includes a spring-loaded tip seal mechanism, a filter membrane in the ventilation channel, a venting system with a silicone membrane, a permeable membrane, and / or a silicone membrane to reduce contamination of the composition contained within the dispenser. In some embodiments, the multi-use dispenser employs a spring-loaded tip seal mechanism, thereby preventing microorganisms from entering the device during a spraying event. In some embodiments, the multi-use dispenser employs a metal-free fluid path, thereby preventing oxidation of the formulation.

[0098] Instead of using preservatives, advanced spray dispenser technology represents an alternative way to keep nasal sprays sterile by preventing bacteria from entering and contaminating the drug formulation. Such dispensers may also be employed in the present invention, thereby further reducing the microbial load in multi-use dispensers. As described herein and known to those skilled in the art, for example from Nemera or Aptar, the antimicrobial properties of ALPHA-1062 or its salts, when combined with "preservative-free" dispenser technology, results in an unexpectedly good shelf life (either during storage or during use) of liquid compositions containing ALPHA-1062 or its salts.

[0099] In one embodiment, the dispenser is configured for multiple individual spray events of 5-1000 μL, preferably 5-500 μL, more preferably 10-300 μL, more preferably 20-200 μL.

[0100] In one embodiment, the dispenser is configured for a plurality of individual spray events of a volume of about 5 μL, or 10 μL, 15 μL, 20 μL, 25 μL, 30 μL, 35 μL, 40 μL, 45 μL, 50 μL, 55 μL, 60 μL, 65 μL, 70 μL, 75 μL, 80 μL, 85 μL, 90 μL, 95 μL, 100 μL, 120 μL, 140 μL, 160 μL, 180 μL, 200 μL, 250 μL, 300 μL, 350 μL, 400 μL, 450 μL, or 500 μL. Ranges consisting of any given value of the aforementioned values ​​are also contemplated.

[0101] In one embodiment, the dispenser contains a total volume of the composition of from 1 to 500 mL, preferably from 1 to 100 mL, more preferably from 2 to 50 mL, such as about 5 mL, 10 mL, or 15 mL.

[0102] In one embodiment, the dispenser comprises a volume of about 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 11 mL, 12 mL, 13 mL, 14 mL, 15 mL, 16 mL, 18 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, 110 mL, 120 mL, 130 mL, 140 mL, 150 mL, 160 mL, 170 mL, 180 mL, 190 mL, 200 mL, 220 mL, 240 mL, 260 mL, 280 mL, 300 mL, 350 mL, 400 mL, 450 mL, or 500 mL. Ranges consisting of any given value of the aforementioned values ​​are also contemplated.

[0103] In some embodiments, the dispenser should contain a sufficient amount of the composition to administer multiple doses. In some embodiments, the dispenser contains a volume of the composition sufficient for at least 2, or at least or about 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, or about 100 individual doses or more. Ranges formed from any given value of the preceding values ​​are also contemplated.

[0104] In one embodiment, the dispenser is configured for multiple individual spray events of 20-200 μL, and the dispenser contains a total volume of solution of 2-50 mL.

[0105] In one embodiment, the multi-use dispenser is configured and / or the ALPHA-1062 or salt thereof is administered in a dose of 1-100 mg, 1-3 times per day, preferably over multiple days.

[0106] In one embodiment, the multi-use dispenser is configured and / or administered with ALPHA-1062 or a salt thereof at a dose of 2-40 mg, twice daily, preferably over multiple days.

[0107] In one embodiment, the multi-use dispenser is configured to administer ALPHA-1062 or a salt thereof in a dosage of 0.1-200 mg, in a dosage of 1-100 mg, preferably in a dosage of 2-40 mg, preferably 1-3 times per day, more preferably twice per day, even more preferably once per day, over multiple days.

[0108] In one embodiment, ALPHA-1062 or a salt thereof is administered intranasally as a 2-40% weight / volume (w / v) solution in a volume of 20-100 microliters, 1-3 times per day for multiple days in each of multiple nasal spray events.

[0109] In one embodiment, the multi-use dispenser is configured and / or administered with ALPHA-1062 or a salt thereof, preferably intranasally, as about a 10% weight / volume (w / v) solution in an amount of about 50 microliters, one to three times daily, preferably over multiple days, in each of multiple administration events.

[0110] In one embodiment, the multi-use dispenser is configured and / or administers ALPHA-1062 or a salt thereof by intranasal, buccal, or sublingual administration, preferably as a 2-40% weight / volume (w / v) solution, e.g., in volumes of 20-100 microliters, preferably in multiple (intranasal or oral (sublingual / buccal)) administration events, e.g., 1-3 times per day, preferably over multiple days.

[0111] In some embodiments, liquid formulations are configured for any one or more of the above administration modes.Those skilled in the art are aware of the technical measures that are adopted when configuring a composition for a particular administration mode.For example, a composition configured for nasal administration may be formulated, packaged, or prepared in a different manner than a composition prepared for oral administration.

[0112] In embodiments of the invention, TBI is classified as mild TBI, moderate TBI, or severe TBI.

[0113] In relation to the use of ALPHA-1062 or a salt thereof in the treatment of confirmed or suspected TBI, the composition or compound may be administered to a subject immediately following an acute TBI event, as well as to a subject who has previously had a suspected or confirmed TBI event. Thus, the use of ALPHA-1062 or a salt thereof, in some embodiments, refers to the treatment and / or prevention of acute and / or chronic symptoms and / or outcomes of TBI.

[0114] Thus, in embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject having or suspected of having a TBI, wherein the subject exhibits one or more symptoms selected from the group consisting of dizziness, balance problems, headache, nausea, vomiting, light sensitivity, memory impairment, sleep abnormalities, poor concentration, and visual impairment.

[0115] In embodiments, the present invention also relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, for use in treating a confirmed or suspected traumatic brain injury (TBI) in a subject having or suspected of having moderate or severe TBI, wherein the subject exhibits one or more symptoms selected from the group consisting of weakness in the arms and legs, balance and coordination problems, severe or increasingly severe headaches, impaired sensory perception, impaired cognitive abilities, impaired memory, impaired communication and learning, personality changes, behavioral abnormalities, and impaired vision and hearing.

[0116] In embodiments, the present invention also relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, for use in treating a confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the subject has or is suspected of having a severe TBI and exhibits one or more symptoms selected from the group consisting of paralysis, coma, fainting, dilated pupils, loss of cerebrospinal fluid from the ears or nose, loss of bowel and / or bladder control, breathing problems, slow pulse, breathing problems, slow breathing rate with elevated blood pressure, and ptosis or facial weakness.

[0117] In embodiments, the present invention also relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, for use in treating a subject with confirmed or suspected traumatic brain injury (TBI), the traumatic brain injury (TBI) resulting in or involving one or more injuries selected from the group consisting of hematoma, contusion, intracerebral hemorrhage, subarachnoid hemorrhage, diffuse injury, diffuse axonal injury, ischemia, primary brain injury, and secondary brain injury.

[0118] The formulations and compounds according to the present invention show surprising beneficial effects in the treatment of TBI or other potential brain trauma, reducing the severity of TBI and / or preventing (reducing the risk of) the occurrence of short-term and long-term symptoms of TBI. As shown in the examples herein, transmucosal administration of a composition comprising ALPHA-1062 or a salt thereof according to the present invention to a subject with TBI or other brain trauma results in improved recovery of motor skills, memory function, and other cognitive functions when compared to vehicle treatment after TBI.

[0119] In embodiments, the traumatic brain injury (TBI) is caused by a closed head injury.

[0120] In embodiments, the traumatic brain injury (TBI) is caused by a penetrating head injury.

[0121] In some embodiments, the traumatic brain injury (TBI) is caused by a head injury resulting from an accident selected from the group consisting of a fall, a motor vehicle-related accident, a blow or blow to the head from or against an object, a sports-related accident, interpersonal physical violence or violence by other means. In some embodiments, the traumatic brain injury (TBI) is caused by an incident and / or accident occurring in a contact sport or contact sports such as American football, rugby, soccer, martial arts, Australian rules football, hockey, basketball, etc. TBI may also be caused in some embodiments by a sports accident induced by being hit by a ball, for example in baseball or cricket, or other sports.

[0122] In some embodiments, the traumatic brain injury (TBI) is a blast-related TBI or military TBI, which may occur in civilians or military personnel, for example, due to an explosion near the subject. Head injuries caused by bullets, violent impacts, or shock waves from explosive weapons are the main cause of military traumatic brain injury (TBI), a recognized medical neurological condition that affects many military personnel around the world. Head injuries from exposure to blast waves as a result of military conflicts are a major health problem that is becoming increasingly serious, especially among military service personnel. Co-morbidities often associated are post-traumatic stress disorder, depression, anxiety, sleep disorders, attention disorders, and cognitive disorders.

[0123] In embodiments, the subject having or suspected of having a TBI is an infant under the age of 4.

[0124] In some embodiments, the subject is under 12 months of age.

[0125] In embodiments, the subject having or suspected of having a TBI is a child between the ages of 4 and 12. In some embodiments, the subject is an adolescent between the ages of 12 and 17.

[0126] In embodiments, the subject having or suspected of having a TBI is an adult selected from adults between the ages of 18 and 65.

[0127] In some embodiments, the subject is an adult over 65 years of age (elderly).

[0128] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the traumatic brain injury (TBI) is associated with trauma to the nasal cavity.

[0129] In embodiments, the present invention relates to the treatment of traumatic brain injury (TBI) and the combined treatment and / or prevention of microbial infection of the nasal cavity.

[0130] In embodiments, the present invention also relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the traumatic brain injury (TBI) is associated with disruption of the blood-brain barrier.

[0131] In embodiments, the present invention relates to the treatment of traumatic brain injury (TBI) and the concomitant treatment and / or prevention of microbial infections of the central nervous system.

[0132] The combination of therapeutic properties of ALPHA-1062, including both neurological brain function improving properties and antibacterial properties, is an unexpected combination of benefits that makes the present invention ideally suited for TBI treatment, especially in situations where first responders or emergency medical personnel need to treat quickly after injury, but also relevant for continued use during recovery. In particular, any brain trauma combined with injury to the nasal or oral cavity can be effectively treated with this combination of properties.

[0133] Accordingly, the present invention also relates to a method described herein for treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the subject is additionally suffering from a microbial infection, the method comprising administering ALPHA-1062 or a salt thereof to a subject in need thereof.

[0134] ALPHA-1062 exhibits strong antibacterial effects against a variety of pathogenic microorganisms. As such, the compound is believed to be ideally suited for treating TBI in a subject, the subject additionally having a microbial infection. In some embodiments, the microbial infection is associated with TBI. In some embodiments, the infection is unrelated to TBI. In some embodiments, the infection is a microbial infection, and the microbial infection is preferably a pathogenic microbial infection in the subject.

[0135] According to the preferred modes of administration described herein, the compounds contact various mucosal surfaces that may be subject to unwanted infection, i.e. infection by pathogens. Thus, the antibacterial activity of the compounds provides a beneficial effect upon administration to a subject.

[0136] In embodiments, the present invention also relates to a method of reducing the incidence of traumatic brain injury (TBI) in an individual or treating traumatic brain injury (TBI) in an individual, comprising administering to the individual an effective amount of the compound ALPHA-1062 or a salt thereof.

[0137] In embodiments, the present invention also relates to a method of reducing the incidence of a symptom selected from the group consisting of dizziness, balance problems, headaches, nausea, vomiting, light sensitivity, memory problems, sleep abnormalities, poor concentration, and visual impairment as a result of traumatic brain injury in a subject, or treating traumatic brain injury in a subject, comprising administering to the subject an effective amount of a compound of ALPHA-1062 or a salt thereof.

[0138] In embodiments, the present invention also relates to a method of reducing the incidence of symptoms selected from the group consisting of arm and leg weakness, balance and coordination problems, severe or increasing headaches, impaired sensory perception, impaired cognitive abilities, impaired memory, impaired communication and learning, personality changes, behavioral abnormalities, and impaired vision and hearing as a result of traumatic brain injury in a subject, or treating traumatic brain injury in a subject, comprising administering to the subject an effective amount of a compound of ALPHA-1062 or a salt thereof.

[0139] In embodiments, the present invention relates to a method of reducing the incidence of a symptom selected from the group consisting of paralysis, coma, fainting, dilated pupils, loss of cerebrospinal fluid from the ears or nose, loss of bowel and / or bladder control, breathing problems, slow pulse, breathing problems, slow breathing rate with elevated blood pressure, and ptosis or facial weakness as a result of traumatic brain injury in a subject, or treating traumatic brain injury in a subject, comprising administering to the subject an effective amount of a compound of ALPHA-1062 or a salt thereof.

[0140] In some embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof and preferably one or more pharma- ceutically acceptable carriers for use in the treatment of confirmed or suspected traumatic brain injury (TBI) in a subject, characterized in that the pharmaceutical composition is suitable for intranasal, oral, and / or sublingual application.Accordingly, the present invention relates to a nasal drop, nasal spray, or sublingual drop in the form of a liquid composition for transmucosal administration via the nasal or oral mucosa.

[0141] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject by transmucosal administration, the composition being an aqueous solution comprising 2-40% weight / volume, preferably 5-15% weight / volume, more preferably 10% weight / volume (w / v) of the chemical entity.

[0142] In some embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof for use in treating a confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the compound ALPHA-1062 or a salt thereof is present in a concentration of 1-500 mg / mL, preferably in a concentration of 1-200 mg / mL, more preferably in a concentration of 5-200 mg / mL, or in a concentration of 5-100 mg / mL. These dosage ranges are provided without limiting the scope of the invention. In other embodiments, the effective dose may also be greater than the effective dose that can be delivered in 100 microliters at 200 milligrams per mL of solution.

[0143] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the compound ALPHA-1062 is present as a gluconate salt, preferably in a concentration of 50-100 mg / mL.

[0144] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the composition is administered at a dose of 0.1-200 mg, preferably at a dose of 1-100 mg, more preferably at a dose of 2-40 mg, 1-3 times daily.

[0145] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, the composition being administered intranasally as a 2-40% weight / volume (w / v) solution of ALPHA-1062 in a volume of 20-100 microliters, preferably in a single spray event, preferably 1-3 times daily, preferably for 5-30 days following the TBI.

[0146] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, for use as a medicament for stimulating and / or enhancing neurogenesis and / or neuronal recovery in the brain of a subject confirmed or suspected of having a traumatic brain injury (TBI).

[0147] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof for use as a medicament for preventing, inhibiting, and / or reducing neuronal cell loss in the brain of a subject confirmed or suspected of having a traumatic brain injury (TBI).

[0148] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, for use as a medicament for reducing the size of one or more lesions and / or damage in the brain of a subject confirmed or suspected of having a traumatic brain injury (TBI).

[0149] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, for use as a medicament for the preservation of brain tissue and / or neuronal tissue in the brain of a subject confirmed or suspected of having a traumatic brain injury (TBI).

[0150] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, for use as a medicament for reducing and / or preventing the level and / or formation of pathological p-Tau in the brain of a subject confirmed or suspected of having a traumatic brain injury (TBI).

[0151] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof for use as a medicament for preventing, reducing the risk, and / or delaying or reducing the onset of neurodegeneration in a subject confirmed or suspected of having a traumatic brain injury (TBI).

[0152] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof for use as a medicament in preventing, reducing the risk of, and / or delaying or reducing the onset of neurodegeneration in a subject confirmed or suspected of having a traumatic brain injury (TBI), wherein ALPHA-1062 is administered during the acute phase of TBI or within the first 6 months after a TBI or suspected TBI, preferably within 2 months, 1 month, 3 weeks, 2 weeks, 1 week, or within 6, 5, 4, 3, or 2 days after a TBI or suspected TBI.

[0153] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof for use as a medicament for preventing, reducing the risk, and / or delaying or reducing the onset of neurodegeneration in a subject confirmed or suspected of having a traumatic brain injury (TBI) by preventing, reducing the levels of, and / or delaying or reducing the onset of pathological p-Tau.

[0154] In some of the above-described embodiments, the confirmed or suspected traumatic brain injury (TBI) is a moderate traumatic brain injury. In some of the above-described embodiments, the confirmed or suspected traumatic brain injury (TBI) is a mild traumatic brain injury.

[0155] In a preferred embodiment, ALPHA-1062 or a salt thereof is administered acutely after TBI, for example within 48 hours, more preferably within 24 hours, more preferably within 12 hours, 10 hours, 8 hours, 6 hours, 4 hours, or 2 hours after TBI, this period is sometimes referred to as the acute phase of TBI.

[0156] In embodiments, the composition is administered to the oral cavity as a 2-40% weight / volume (w / v) solution of ALPHA-1062 in a volume of 20-100 microliters, preferably in a single spray event, preferably 1-3 times daily, preferably for 5-30 days following TBI.

[0157] In embodiments, the present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the composition is administered intranasally, orally, or sublingually as about a 10% weight / volume (w / v) solution in a volume of 20-100 microliters, for example about 50 microliters, in 1-3 administration events, preferably a single administration event, twice daily, preferably over multiple days.

[0158] In several embodiments, the pharmaceutical composition is an aqueous solution containing 2-20% weight / volume (w / v), preferably 5-15% weight / volume (w / v), and more preferably 10% weight / volume (w / v) of the chemical.

[0159] To be suitable for transmucosal delivery in the oral or nasal cavity, in some embodiments the compositions are formulated as a concentrated saline solution, or as an emulsion, or as a selfmicroemulsifying drug delivery system (SMEDD), or as a micronized powder formulation.

[0160] In embodiments, the dosing regimens described herein are employed using a multi-use dispenser, as described herein, and any feature disclosed with respect to the dispenser is also deemed to be disclosed with respect to the dosing regimen, and vice versa.

[0161] In some embodiments, the concentration of ALPHA-1062 or a salt thereof preferably employed in the above-mentioned dosing regimen is preferably about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% by weight or more of the solution or emulsion. Dosing regimens disclosing the administration of about 10% by weight, preferably 10%, of the solution or emulsion may alternatively be employed at any of the above-mentioned concentrations or values ​​similar thereto. Concentration ranges consisting of any given value of the above-mentioned values ​​are also contemplated.

[0162] In some embodiments, it is contemplated that ALPHA-1062 or a salt thereof is used in a manner that allows distribution of the substance in a patient after administration with a brain-to-blood concentration ratio of greater than 5, preferably greater than 10, and more preferably between 15-25.

[0163] The present invention also relates to a pharmaceutical composition comprising the compound ALPHA-1062, or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, wherein the composition is administered within 24 hours of the injury.

[0164] In another embodiment, the composition is administered within one hour of the injury.

[0165] In embodiments, administration with as little delay as possible after TBI is preferred, for example, within 1 week, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day of the TBI, hi embodiments, administration is performed within 24 hours, 12 hours, 8 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, or 1 hour of the TBI.

[0166] In some embodiments, administration following TBI is performed within 1 week, 2 weeks, or 3 weeks, 1 month, or 1 year of TBI. In some embodiments, administration is performed within 48 months, 36 months, 24 months, 12 months, 6 months, 5 months, 4 months, 3 months, 2 months, or 1 month of TBI. Preferably, administration of ALPHA-1062 during the acute phase of TBI or immediately after TBI, for example within 3 months, 2 months, or 1 month of TBI, shows beneficial effects.

[0167] The dosing regimen described herein according to some embodiments represents a novel and surprisingly beneficial advancement in comparison with the prior art with respect to effective galantamine treatment. The biological and medical effects of galantamine have never been tested before with respect to the potential effects caused by administration at high doses. Many patients who require galantamine treatment could not be treated due to the significant side effects caused by effective doses of galantamine. To obtain a meaningful level of galantamine in the subject's brain, the prior art teaches high, but often severely intolerable, doses. Since only a small portion of orally or intranasally administered galantamine drug reaches the brain, the dose required to be effective during the treatment of brain diseases is often intolerably high due to the large amount of galantamine in other tissues of the body, thereby causing undesirable side effects.

[0168] The multiple embodiments and features of the invention described with respect to the pharmaceutical compositions, substances and salts thereof, multi-use dispensers, and various methods described herein are deemed to be disclosed with respect to any and all other aspects of the disclosure, such that features that characterize a method or dispenser may be employed to characterize a composition or substance, and vice versa. The various aspects of the invention are unified by, benefit from, are based on, and / or are related by the general and surprising discovery of the beneficial and curative effects of the use of ALPHA-1062 in the treatment of TBI, particularly when administered via a transmucosal route. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0169] The present invention relates to a pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof, or the compound ALPHA-1062 or a salt thereof, for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject. The present invention also relates to single-use or multi-use dispensers configured for intranasal or transmucosal administration of the compounds or compositions of the invention, as well as various methods of treating TBI, comprising administering a composition comprising ALPHA-1062 or a salt thereof.

[0170] The preferred molecule of the present invention is ALPHA-1062, also known as GLN-1062 or Memogain®. In the examples of the present invention and in one preferred embodiment of the present invention, the form adopted is the gluconate salt of ALPHA-1062 (ALPHA-1062 gluconate). For completeness, the compound ALPHA-1062 is a galantamine prodrug, which shows no or negligible activity as a cholinesterase inhibitor or nicotinic modulator before cleavage. Upon esterase cleavage, active galantamine is released.

[0171] However, the terms "active agent" or "active pharmaceutical ingredient" (API) may be used for ALPHA-1062, as it is a preferred compound of the invention in some embodiments. In other embodiments, the compound galantamine may also be considered an active agent or related drug molecule.

[0172] The chemical name (IUPAC) of ALPHA-1062 is (4aS,6R,8aS)-5,6,9,10,11,12-hexahydro-3-methoxy-11-methyl-4aH-[1]benzofuro[3a,3,2-ef][2]benzazepin-6-ol benzoate.

[0173] Molecular formula of free base: C 24 H 25 NO 4;The molecular formula of gluconic acid is C 6 H 12 O 7 ;Molecular weight of free base: 391.47 g / mol;Molecular weight of ALPHA-1062 gluconate: 587.61 g / mol;Conversion factor: 1 mg base = 1.501 mg salt.

[0174] Chemical structure of ALPHA-1062: [ka] Galantamine is C 17 H 21 NO 3 and 287.359 g mol -1 and the following structure: [ka] The chemical structure of ALPHA-1062 gluconate is as follows: [ka] As an example, the gluconate salt of ALPHA-1062 can be made according to the following established general scheme: [ka]

[0175] In one embodiment, the composition comprises a crystalline solid form of ALPHA-1062 gluconate (Form A), which has prominent peaks at 3.61, 10.98, 14.41, and 18.44 degrees two-theta (±0.2) in an X-ray powder diffraction pattern.

[0176] These four peaks were selected from the prominent peak list provided below and do not appear to show substantial overlap with the prominent peaks in the XRPD patterns of Forms B-D or Materials E-G, as disclosed in WO 2022 / 150917. Thus, in one embodiment, Form A can be reliably distinguished using one or more prominent peaks when comparing corresponding X-ray powder diffraction patterns, for example, as described above. In one embodiment, the presence of these peaks in the X-ray powder diffraction pattern can be used to identify Form A and / or distinguish Form A from solid forms previously described in the art, such as those described in WO 2014 / 016430.

[0177] In one embodiment, Form A has one or more additional prominent peaks in its powder X-ray diffraction pattern at 15.20, 17.31, 17.79, 22.77, 23.64, 24.88, and 34.31 degrees 2-theta (±0.2), which are selected from the prominent peak list and do not appear to show substantial overlap with the prominent peaks in the XRPD patterns of Forms B through D or Materials E through G.

[0178] In one embodiment, Form A has at least five prominent peaks selected from the list consisting of 3.61, 10.98, 13.80, 14.41, 14.56, 15.08, 15.20, 17.02, 17.31, 17.79, 18.44, 19.24, 20.18, 20.91, 21.22, and 22.40 degrees two-theta (±0.2) in an X-ray powder diffraction pattern.

[0179] Typically, it is not necessary to detect all peaks from this list to determine the presence of Form A in any given preparation. According to the present invention, for example, in some embodiments, one, two, three, four, five, six, seven, eight, nine, ten or more peaks, preferably those with relatively high signal intensity, may be taken to determine any given crystalline form. For example, the four, five, six, seven, eight, nine or ten most intense peaks may be taken to identify any given crystalline form. In one embodiment, sufficient identification of any given crystalline form, such as Form A, is achieved when the presence of at least three or four prominent peaks can be determined based on XRPD comparison.

[0180] Typically, a prominent XRPD peak is the most intense low-angle non-overlapping peak observed in an XRPD pattern. In some embodiments, a "prominent peak" has a relative intensity of preferably 20% or more, preferably 30% or more, more preferably 40% or more in a powder X-ray diffraction pattern. However, the relative intensity value may vary depending on the device or analytical mode, and is not inherently limiting of the solid forms described herein.

[0181] In one embodiment, Form A has peaks at 7.25 and / or 12.67 degrees 2-theta (±0.2) in its powder X-ray diffraction pattern. These peaks are of relatively low intensity compared to the peaks outlined above as the major peaks. However, the peaks at 7.25 degrees 2-theta and / or 12.67 degrees 2-theta appear to be absent from all other patterns of Forms B through D or Materials E through G.

[0182] In one embodiment, the peaks are determined using powder X-ray diffraction analysis in transmission mode.

[0183] In one embodiment, Form A has at least three peaks in an X-ray powder diffraction pattern selected from the list consisting of 10.98, 14.41, 17.31, 18.44, and 22.40 degrees two-theta (±0.2). In one embodiment, the three peaks are within the five peaks with the highest relative intensity in an X-ray powder diffraction pattern obtained using analysis in transmission mode. In one embodiment, these five peaks are the most intense peaks in an XRPD pattern using transmission mode, as outlined in the Examples below.

[0184] In one embodiment, the peaks are determined using powder X-ray diffraction analysis in reflection mode.

[0185] In one embodiment, Form A has at least three peaks in an X-ray powder diffraction pattern selected from the list consisting of 3.61 degrees 2-theta, 7.25 degrees 2-theta, 10.98 degrees 2-theta, 14.56 degrees 2-theta, and 22.40 degrees 2-theta (±0.2). In one embodiment, the three peaks are preferably within the five peaks with the highest relative intensity in an X-ray powder diffraction pattern obtained using analysis in reflection mode. In one embodiment, these five peaks are the most intense peaks in an XRPD pattern using reflection mode, as outlined in the Examples below.

[0186] In one embodiment, Form A has one or more peaks in an X-ray powder diffraction pattern selected from the list consisting of 3.61, 7.25, 10.98, 14.56, 22.40 degrees two-theta (±0.2), which are also observable from an XRPD pattern using reflection mode.

[0187] In one embodiment, Form A has one or more doublets selected from the list consisting of 14.41 and 14.56, 15.08 and 15.20, and 24.88 and 25.09 degrees two-theta (±0.2) in an X-ray powder diffraction pattern. These doublets may be used to identify Form A and, optionally, distinguish the form from other forms.

[0188] A table of typically observed XRPD pattern peaks for Form A collected in transmission mode is provided below.

[0189] Peak List Form A: Peak list determined from the powder X-ray diffraction pattern of Form A according to Figure 8. The accuracy of 2 theta degrees is provided to two decimal places and some variation may be evident depending on the batch or device.

[0190] [Table 1]

[0191] * The peaks may, in some embodiments, be considered prominent peaks observed in an XRPD pattern.

[0192] In one embodiment, Form A exhibits an onset of melting at a temperature between 116-120° C., preferably at a temperature of about 117° C., as assessed using differential scanning calorimetry (DSC).

[0193] In one embodiment, Form A exhibits less than 1% weight loss, preferably less than 0.5%, more preferably less than 0.3% or less than 0.2%, prior to the onset of melting using DSC, as assessed using Thermo-Gravimetric Analysis (TGA).

[0194] As used herein, crystalline preferably refers to a material that has an ordered long-range molecular structure. The crystallinity of a crystalline form can be determined by a number of techniques, including, for example, powder X-ray diffraction, moisture sorption, differential scanning calorimetry, solution calorimetry, and dissolution properties.

[0195] Crystalline organic compounds consist of a large number of atoms arranged in a periodic array in three-dimensional space. Structural periodicity usually exhibits obvious physical characteristics such as sharp and well-defined spectral signatures by most spectroscopic probes (e.g., X-ray diffraction, infrared, and solid-state NMR). X-ray diffraction (XRD) is recognized as one of the most sensitive methods for determining the crystallinity of solids. Crystals produce well-defined diffraction maxima that occur at specific angles that coincide with lattice spacings, as predicted by Bragg's law. In contrast, amorphous materials do not have long-range order. They often retain additional volume between molecules, as in the liquid state. Amorphous solids usually reveal featureless XRD patterns with broad diffuse halos, since there is no long-range order of a repeating crystal lattice.

[0196] Crystalline forms are preferred in many pharmaceutical applications. Crystalline forms are generally more thermodynamically stable than amorphous forms of the same substance. This thermodynamic stability is preferably reflected in the improved physical stability of the crystalline form. The ordered packing of molecules in crystalline solids preferably resists the incorporation of chemical impurities. Thus, crystalline materials generally have a higher chemical purity than their amorphous counterparts. The packing in crystalline solids generally confines the molecules to well-defined lattice positions, reducing the molecular mobility that is a prerequisite for chemical reactions. Thus, with few notable exceptions, crystalline solids are more chemically stable than amorphous solids of the same molecular composition. Preferably, the crystalline forms of ALPHA-1062 gluconate disclosed in the present application have one or more of the advantageous chemical and / or physical properties disclosed herein.

[0197] As used herein, the term stable can refer to either chemical stability or polymorphic stability. Polymorphic stability refers to the ability of a polymorphic form to remain in its particular crystalline state under suitable storage conditions. For example, a stable polymorphic form maintains at least about 95% by weight, preferably at least about 98% by weight, more preferably at least about 99% by weight or more of the crystalline form, in other words, the form remains unchanged after storage under the indicated conditions for the indicated time. In the context of the present invention, ALPHA-1062 gluconate of Form A can be, for example, stored under conditions at room temperature and at low water activity, for example, below about 43% RH or 0.12a. w It is believed that Form A exhibits good stability for multiple months under conditions such as less than 1000 mol / L. In some embodiments, Form A exhibits good chemical stability. In other words, ALPHA-1062 gluconate as Form A exhibits low, negligible or no conversion to different chemical structures after storage under appropriate conditions.

[0198] Powder X-ray diffraction (PXRD) measures the diffraction pattern of crystalline materials. Each active pharmaceutical ingredient (API) produces a specific pattern depending on the structure of its crystal lattice. Each polymorph, pseudopolymorph, polymorphic salt, or co-crystalline material has its own specific pattern. For this reason, PXRD of an API can be performed under controlled conditions to assess the presence or absence of crystalline material and any form transformations.

[0199] PXRD can also be used to determine whether a change in crystalline form has occurred in a drug product, for example, during storage or stability studies. Thus, the identification of a crystalline form depends on the presence of detectable diffraction peaks for any given crystalline form. In addition, the API peaks must be distinguishable from any crystalline excipient peaks, and the composition should be evaluated after formulation. PXRD can also be used as a qualitative and possibly quantitative assessment of the crystallinity of a pure API. Those skilled in the art can evaluate a PXRD pattern and identify the presence and / or absence of suitable peaks that can be employed to characterize any given crystalline form of an API without undue effort.

[0200] In some embodiments, the peaks determined by PXRD analysis are essentially the same as those presented in the following examples.The term "essentially the same" in reference to PXRD means that the variability of peak positions and relative intensities of peaks is taken into account.For example, the typical accuracy of 2-theta values ​​is within ±0.2° 2-theta.

[0201] As used herein, characteristic XRPD peaks are a representative subset of peaks from the XRPD pattern of a crystalline form of a material that can be statistically proven to be distinct from other crystalline forms of the material. Not all crystalline polymorphs of a material necessarily have characteristic peaks.

[0202] As used herein, a prominent XRPD peak is typically the most intense low angle non-overlapping peak observed in an XRPD pattern. In some embodiments, a "prominent peak" preferably has a relative intensity of 20% or greater, preferably a relative intensity of 30% or greater, more preferably a relative intensity of 40% or greater in a powder X-ray diffraction pattern.

[0203] As used herein, a representative XRPD peak is a peak from the XRPD pattern of a crystalline form of a material that statistically shows no deviation from particle size / shape or preferred orientation during replicate samples and measurements.

[0204] As used herein, preferred orientation is a phenomenon observed in XRPD analysis where, depending on the size / shape of the particles and the pattern collection technique employed, it is extremely difficult or impossible to randomly orient the particles of a material during collection to achieve a pattern with statistically consistent intensity.

[0205] With regard to the relative intensities and prominent peaks of the powder X-ray diffraction patterns mentioned above, the provided values ​​of relative intensities are not intended as limiting the identification of the prominent or characteristic peaks mentioned. As known to those skilled in the art, relative peak intensities exhibit some instrumental variability, batch-to-batch variability, and variability due to degree of crystallinity, preferred orientation, sample preparation, and are therefore provided only as an indication and qualitative measure of the intensity of the peaks in the powder X-ray diffraction pattern, but are not intended as a limiting definition.

[0206] Thus, the term "prominent peak" in the context of defining the present invention is not limited to the respective relative intensities provided above, and any one or more of the respective peaks may be determined as the prominent peak of any given form of ALPHA-1062 gluconate. Preferably, at least one, two, three, or four prominent peaks are used to characterize the crystalline form, and in other embodiments, at least five, six, seven, eight, nine, or ten prominent peaks may be employed. Thus, the prominent peaks are also not limited to peaks unique to any given crystalline form, rather, the peaks, optionally in combination with some other peaks from the PXRD pattern, may be used to identify the crystalline form. In the context of the present invention, crystalline forms A to D may share multiple prominent peaks, but may also exhibit peaks that are different from each other that may be used to distinguish any two forms. In some embodiments, the prominent peaks referred to in the embodiments of the present invention may also be characteristic and / or representative peaks. As used herein, the term "self-preserving" is a description of the antimicrobial properties of a compound, i.e., ALPHA-1062 or a salt thereof, or a composition comprising such an agent, that does not require the presence of an additional antimicrobial preservative.

[0207] In a preferred embodiment, the self-preserving liquid composition maintains the absence of viable microorganisms present in the composition, has a low or negligible number, or has a relatively slow growth rate or reduces the number of viable microorganisms in the composition. In some embodiments, the "self-preserving" property indicates that the rate of microbial growth (cell growth or division) in the composition over time is slower than in the absence of the relevant compound (ALPHA-1062). Thus, as used herein, a "self-preserving" liquid composition exhibits a lower number of viable microorganisms compared to a composition without such "self-preserving" property. In some embodiments, a "self-preserving" composition does not contain an additional antimicrobial preservative and exhibits no increase or a negligible increase in the number of viable microorganisms in the composition over an extended period of time, preferably at least 14 days or at least 28 days.

[0208] As used herein, the term "antimicrobial" describes the property of a compound or composition that reduces or exhibits no or negligible increase in the number of viable microorganisms in the composition for at least 14 days or at least 28 days. In some embodiments of the present invention, pathogenic fungi, gram-negative bacteria and / or gram-positive bacteria can be killed or inhibited from growing by ALPHA-1062 or its salts. In some embodiments, the term "antimicrobial" is defined according to the guidelines set forth for preservatives in the USP 51 test. Thus, the term may depend on the type of microorganism and the duration of the test. The concentration of added antimicrobial preservatives is usually kept to a minimum or avoided entirely, especially when the active ingredient of the formulation has inherent antimicrobial activity, as in the case of ALPHA-1062 or its salts. Antimicrobial efficacy, whether inherent to the product or generated from the addition of antimicrobial preservatives, generally needs to be demonstrated for multi-dose topical and oral dosage forms, as well as other dosage forms such as ophthalmic solutions, ocular solutions, nasal solutions, irrigation solutions, and dialysis solutions. As used herein, the agent ALPHA-1062 exhibits antimicrobial preservative properties that are inherent to the molecule, and therefore necessitates little or no additional use of additional preservatives in the compositions of the present invention.

[0209] "Traumatic brain injury" (TBI) is a disruption of the normal function of the brain that can be caused by a blow, collision, or impact to the head, a sudden, violent strike of an object against the head, or when an object penetrates the skull and enters brain tissue. Observation of one of the following clinical signs constitutes an alteration of normal brain function: loss of consciousness or reduced consciousness, loss of memory of events before and after the event (amnesia), focal neurological deficits such as muscle weakness, loss of vision, changes in speech, changes in mental status such as disorientation, slowed thinking or difficulty concentrating.

[0210] Symptoms of TBI can be mild, moderate, or severe, depending on the extent of the injury to the brain. Mild cases may result in a brief alteration in mental state or consciousness. Severe cases may result in prolonged unconsciousness, coma, or even death.

[0211] Symptoms of TBI can vary greatly depending on the severity of the head injury and can include vomiting, lethargy, headache, confusion, paralysis, coma, unconsciousness, dilated pupils, visual disturbances (blurred or double vision, intolerance to light, loss of eye movement, blindness), dizziness and imbalance, difficulty swallowing, numbness or tingling, drooping eyelids or facial weakness, and loss of bowel or bladder control.

[0212] Short-term and long-term symptoms of TBI may further include one or more symptoms selected from the group including dizziness, balance problems, headache, nausea, vomiting, light sensitivity, memory impairment, sleep abnormalities, poor concentration and vision problems, weakness in the arms and legs, balance and coordination problems, severe or increasing headaches, impaired sensory perception, impaired cognitive abilities, memory impairment, impaired communication and learning, personality changes, behavioral abnormalities, impaired vision and hearing, paralysis, coma, fainting, dilated pupils, loss of cerebrospinal fluid from the ears or nose, loss of bowel and / or bladder control, breathing problems, slow pulse, breathing problems, slow breathing rate with increased blood pressure, ptosis or facial weakness, hematoma, contusion, intracerebral hemorrhage, subarachnoid hemorrhage, diffuse injury, diffuse axonal injury, ischemia, primary brain injury, and / or secondary brain injury.

[0213] As used herein, "primary brain injury", i.e., skull fracture, focal injury, or diffuse axonal injury, occurs at the time of the initial brain injury, i.e., TBI event, which cannot be prevented, but is only addressed by subsequent treatment. One goal of treating initial brain injury is to prevent or further reduce "secondary brain injury". The term "secondary brain injury" typically refers to changes that progress within minutes, hours, days, or even months after the initial injury. Secondary brain injury includes the entire series of events or stages of cellular, chemical, tissue, or vascular changes in the brain that contribute to further destruction of brain tissue.

[0214] TBI can cause contusions and "mass lesions" in areas of focal injury such as contusions that increase pressure within the brain. Secondary injuries caused by TBI or associated TBI can include one or more of the following injuries: hematoma, (brain) contusion, intracerebral hemorrhage, subarachnoid hemorrhage, diffuse injury, diffuse axonal injury, ischemia, and / or skull fractures.

[0215] In embodiments of the present invention, "hematoma" refers to a blood clot in or on the surface of the brain, which may appear anywhere in the brain. An epidural hematoma is a collection of blood between the dura mater (the protective covering of the brain) and the inside of the skull. A subdural hematoma is a collection of blood between the dura mater and the arachnoid layer located on the surface of the brain.

[0216] In embodiments of the present invention, "contusion" or "cerebral contusion" may refer to a contusion of brain tissue that is pathologically comparable to a contusion in other parts of the body. A contusion consists of an area of ​​damaged or swollen brain mixed with blood that has leaked from arteries, veins, or capillaries. The most common contusion is at the front base of the brain, but contusions can appear anywhere.

[0217] In embodiments of the present invention, "intracerebral hemorrhage" (ICH) refers to bleeding in brain tissue that may be associated with other brain injuries, particularly contusions, which may be surgically removed depending on the size and location of the hemorrhage.

[0218] In embodiments of the present invention, "subarachnoid hemorrhage" (SAH) is caused by bleeding into the subarachnoid space. It appears as a thin, diffuse blood spread over the surface of the brain and is common after TBI. Most cases of SAH associated with head trauma are mild. Hydrocephalus can result from severe traumatic SAH.

[0219] In embodiments of the present invention, "diffuse injury" describes microscopic changes scattered throughout the brain that do not show up on a CT scan and can occur as a result of TBI. This category of injury, known as diffuse brain injury, can occur with or without an associated mass lesion.

[0220] In some embodiments of the present invention, "diffuse axonal injury" refers to the dysfunction and gradual loss of axons. These long extensions of nerve cells allow them to communicate with each other. If enough axons are damaged in this way, the ability of nerve cells to communicate with each other and integrate their functions can be lost or severely impaired, which can lead to severe disability in the patient.

[0221] In some embodiments of the present invention, "ischemia" describes another type of diffuse injury, or insufficient blood supply to a certain part of the brain. A high percentage of TBI patients experience a reduction in blood supply to very low levels. This is important because the brain that has just suffered a traumatic injury is particularly sensitive to mild reductions in blood flow. Changes in blood pressure during the first few days after head injury can also have adverse effects. Ischemia is also believed to be a major cause of secondary brain injury.

[0222] In embodiments of the present invention, "skull fracture" refers to a linear skull fracture or a simple fracture of the skull that may accompany a TBI.

[0223] Potential forces strong enough to cause a skull fracture can injure the brain underneath. Skull fractures can be alarming if discovered during a patient exam. Fractures at the base of the skull are problematic because they can damage nerves, arteries, or other structures. If the fracture extends into the sinuses, leakage of cerebrospinal fluid from the nose or ears can occur. Depressed skull fractures, in which part of the bone presses on or into the brain, can also occur.

[0224] In some embodiments of the present invention, ALPHA-1062 administration may induce prevention, reduction of risk, reduction of levels, and / or delay or reduction of the onset of pathological p-Tau. As used herein, "p-Tau" refers to phosphorylated Tau protein. Tau proteins include six soluble protein isoforms produced by alternative splicing from the gene MAPT (microtubule associated protein tau). They are primarily responsible for maintaining the stability of axonal microtubules and are abundant in neurons of the central nervous system (CNS). Accumulation of pathologically phosphorylated Tau in neurons is typically associated with neurofibrillary degeneration and is observed in various neurodegenerative diseases. Nervous system pathologies and dementias such as Alzheimer's and Parkinson's disease are associated with phosphorylated Tau protein that has become insoluble aggregates called neurofibrillary tangles. Although the mechanism of tau aggregation has not yet been fully elucidated, several factors, including tau phosphorylation, are thought to be associated with and / or induce this process. Thus, pathological p-Tau is often considered both a marker of neurodegeneration and a causative factor of neurodegeneration.

[0225] "Prodrug" generally describes a drug or compound that is metabolized in the body to a pharmacologically active drug after administration. A prodrug can be administered in place of a corresponding drug compound to improve the absorption, distribution, metabolism, and / or excretion of the corresponding drug. The use of a prodrug with improved bioavailability is particularly advantageous when the corresponding drug is poorly absorbed, for example, via the gastrointestinal route. The administration of a prodrug can reduce adverse or undesired side effects of the corresponding drug and / or improve the bioavailability and / or absorption of the drug.

[0226] A "prodrug" is typically a therapeutically inactive drug that is itself predictably converted to an active metabolite at a specific location in the body. In this sense, a prodrug is an inactive precursor of a parent drug that undergoes conversion to an active drug in vivo by enzymatic cleavage or chemical spontaneous processes in a predictable manner. In the prodrug according to some embodiments discussed herein, there is preferably a covalent ester linkage between the parent drug and the selected promoiety, and upon cleavage of this ester bond, the inactive prodrug releases the active parent drug at or near its target site in the CNS, ideally in the target organ, the brain.

[0227] As used herein, "prodrug" may refer to, but is not limited to, ALPHA-1062 or a salt thereof. In some embodiments of the present invention, administration of the prodrug ALPHA-1062 or a salt thereof is intended to reduce or prevent (partially or entirely) galantamine-related adverse effects and / or improve transmucosal absorption of galantamine compounds.

[0228] Therefore, the goal of some embodiments according to the present invention is to present novel CNS therapeutics for the treatment of TBI with optimal brain bioavailability by being formulated as lipophilic prodrugs and administered via oral or intranasal transmucosal absorption routes.

[0229] The ALPHA-1062 prodrugs disclosed herein are passively transported through the blood-brain barrier (BBB) ​​into the brain. These ALPHA-1062 prodrugs are essentially pharmacologically inactive and therefore do not produce any significant GI or other side effects as long as they remain uncleaved in a particular tissue. After enzymatic cleavage, one molecule of the parent drug is formed from each molecule of the prodrug, thereby resulting in the full pharmacological effect of the drug. If cleavage occurs preferentially in the brain due to enhanced distribution to this organ and the availability of suitable endogenous enzymes therein, a sustained higher concentration of the drug at the target site of the CNS, and thus a greater medically beneficial effect, can be achieved. The preferential transport of the target organ to the brain is further optimized in a surprising and beneficial manner by the transmucosal route of administration in the oral or nasal cavity.

[0230] In summary, these features of the formulations of ALPHA-1062 or salts thereof described in some embodiments herein facilitate sustained and higher concentration delivery of the prodrug to the brain than can be achieved by oral administration of the unmodified drug in tablet form. Improved distribution of the drug to the brain dramatically reduces any side effects that occur locally in the GI tract, thereby allowing for immediate effective administration of the drug at target molecules located in the CNS, such as nicotinic receptors and acetylcholinesterase.

[0231] Since the "blood-brain barrier" (BBB), located at the level of brain capillaries, is the main barrier for the passage of drugs from the blood compartment to the brain, an initial focus on optimizing the penetration of prodrugs through the BBB has yielded promising results. Brain microvascular endothelial cells that form the BBB have typical morphological characteristics, such as tight junctions between cells, lack of fenestrations, and reduced pinocytic activity. Various enzymes further contribute to the restrictive nature of the BBB. The ability of a drug to cross the BBB depends primarily on its physicochemical properties, such as its lipophilicity. As a result, all of the compounds considered in this disclosure are prodrugs with improved lipophilicity compared to their parent compounds.

[0232] "BBCR" should be understood as the brain-to-blood drug concentration ratio after transport equilibrium across the BBB has been achieved.

[0233] The term "transmucosal administration" refers to the transport of a pharmaceutical agent through or across a mucous membrane. Transmucosal routes of administration according to the present invention are defined as intranasal, buccal, and / or sublingual.

[0234] "Nasal or intranasal administration" refers to any application form of a prodrug or its pharmaceutical composition to the nasal cavity. The nasal cavity is covered by a thin mucous membrane that is well vascularized. Therefore, drug molecules can move rapidly across a single epithelial cell layer without first-pass liver and intestinal metabolism. Thus, intranasal administration is used as an alternative to oral administration of, for example, tablets and capsules that result in extensive degradation in the intestine and / or liver.

[0235] "Buccal administration" refers to any form of application that results in absorption across the buccal mucosa, and preferably involves adsorption on the inside of the cheek, on the surfaces of the teeth, or on the gums along the side of the cheek.

[0236] "Sublingual administration" refers to administration under the tongue where the prodrug contacts and diffuses through the mucous membrane beneath the tongue.

[0237] Rapid absorption in the oral cavity is best achieved by sublingual administration, as the mucosal thickness in this region is lower than other oral regions, in addition to being significantly less keratinized (Shojaei A (1998) Buccal mucosa as a route for systemic drug delivery: a review. J Pharm Pharmaceut Sci 1:15-30). Fast dissolving sublingual formulations according to some embodiments of the present invention, such as rapidly disintegrating tablets or liquid-filled capsules, can further help reduce the enzymatic degradation of prodrugs in saliva. The nasal cavity also provides a promising starting point for alternative dosing regimens, with its large surface area, high vasculature, and low enzyme environment. Intranasal delivery according to some embodiments of the present invention can provide high levels of bioavailability similar to intravenous administration, and compared to the latter, has the advantages of non-invasiveness, ease of self-administration, patient comfort, and patient compliance. These advantages may be generally known by those skilled in the art; however, significant obstacles remain to develop such application routes. For long-term systemic delivery, problems of epithelial injury and toxicity need to be resolved, and high concentrations of drug in small volumes of vehicle need to be provided for sufficient bioavailability. This requires the initial selection of suitable chemical compounds that allow the required formulations and concentrations, in addition to finding an appropriate method for administration and developing preferred salts and / or solutions thereof that ultimately allow optimal administration of the active substance to the brain.

[0238] Pharmaceutical compositions suitable for buccal and / or sublingual administration may, in some embodiments, include additional "pharmaceutical acceptable carriers", for example, buccal dosage units may include the active agent to be administered in addition to a polymeric carrier that bioerodes and provides delivery of the active agent over a period of time, and preferably a lubricant such as magnesium stearate. Additional carrier agents are known to those skilled in the art. The active agent may be physically compounded with materials from some or all of the following classes of ingredients that function as pH control agents, preservatives, viscosity control agents, absorption enhancers, stabilizers, solvents, and carrier vehicles. Such agents may, in some embodiments of the present invention, be present in either solid or liquid form of the pharmaceutical composition.

[0239] In some embodiments, a "self-microemulsifying drug delivery system" (SMEDDS) may be present in the pharmaceutical composition, which refers to a drug delivery system that uses microemulsions achieved by chemical means rather than mechanical means; that is, by the inherent properties of the drug formulation rather than by special mixing and handling. This employs the well-known effect exhibited by anethole in many anise-flavored alcoholic beverages. Microemulsions have great potential for use in drug delivery, and SMEDDS (including so-called "U-shaped" microemulsions) are the best of these systems identified so far. SMEDDS are particularly valuable for increasing the absorption of lipophilic drugs taken orally. SMEDDS may include, but are not limited to, formulations of the drug anethole trithione, oridonin, curcumin, vinpocetine, tacrolimus, berberine hydrochloride, nobiletin, and / or piroxicam. Other emulsions are created by mechanical means such as mixing, sonication, vortexing, or homogenization. The first drug marketed as SMEDD was cyclosporine, which had significantly improved bioavailability compared to conventional solutions. SMEDDS offers many advantages: spontaneous formation, ease of manufacture, thermodynamic stability, and improved solubilization of bioactive materials. Improved solubility contributes to faster release rates and greater bioavailability. For many drugs taken orally, faster release rates improve drug acceptance by the recipient. Greater bioavailability means that less drug needs to be used; this can lower costs and also reduces gastric irritation and toxicity of orally taken drugs. For oral use according to some embodiments, SMEDDS may be formulated as a liquid and is applicable for transmucosal administration.

[0240] The pharmaceutical composition may further comprise one or more pharma- ceutically acceptable carriers or excipients in some embodiments. The term "excipient" refers to a pharmacologically inactive component such as a pharmaceutical diluent, disintegrant, carrier, and the like. Excipients useful in the preparation of pharmaceutical compositions are generally safe, non-toxic, and acceptable for veterinary and human pharmaceutical use. Reference to an excipient includes both one excipient and two or more excipients. In some embodiments, excipients are described herein according to "weight %" or "weight percent."

[0241] "Administration" or "treatment" as applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid refers to the contact of a pharmaceutical, therapeutic, diagnostic, compound, or composition to an animal, human, subject, cell, tissue, organ, or biological fluid. "Administration" and "treatment" can refer in some embodiments to, for example, therapeutic, placebo, pharmacokinetic, diagnostic, research, and experimental methods. "Treatment" as applied to a human, veterinary, or research subject refers to therapeutic treatment, prophylactic or preventative measures, research, and diagnostic uses. As used herein, "administer" or "administration" refers to the delivery of a drug or agent of the present invention, or a pharmaceutical composition thereof, to an organism for the purpose of preventing or treating a brain disease associated with cognitive impairment. Suitable routes of administration may include, but are not limited to, oral, rectal, transmucosal, or intestinal administration, or intramuscular, subcutaneous, intramedullary, intrathecal, direct intraventricular, intravenous, intravitreal, intraperitoneal, intranasal, sublingual, buccal, or intraocular injection. A preferred route of administration is transmucosal.

[0242] In some embodiments, liquid formulations are configured for any one or more of the above administration modes.Those skilled in the art will recognize the technical measures that are adopted when configuring a composition for a specific administration mode.For example, a composition that is configured for nasal administration may be formulated, packaged, or prepared in a different manner than a composition that is prepared for oral administration.

[0243] The pharmaceutical composition of the present invention may be prepared in some embodiments by processes well known in the art, for example, by conventional mixing, dissolving, granulating, granulating, dragee-making, levigating, emulsifying, encapsulating, encapsulating, dissolving, or lyophilizing processes. The pharmaceutical composition for use according to the present invention may be formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliaries, which facilitate the processing of the crystals or other forms of the present invention into pharma-ceutical usable preparations in some embodiments. The appropriate formulation depends on the selected route of administration.

[0244] In some embodiments for injection or transmucosal administration, the compounds of the present invention or pharmaceutical compositions thereof may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer. For transmucosal administration according to some embodiments, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0245] In some embodiments for oral administration, the compounds of the present invention or pharmaceutical compositions thereof can be formulated by combining the compounds of the present invention with pharma- ceutically acceptable carriers well known in the art. Such carriers, in some embodiments, allow the compounds of the present invention to be formulated as tablets, pills, lozenges, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like for oral ingestion by patients. Pharmaceutical preparations for oral use can, in some embodiments, be produced by milling the resulting mixture with solid excipients and processing the mixture of granules, optionally after adding other suitable auxiliaries as necessary, to obtain tablets or dragee cores. Useful excipients are in particular fillers such as sugars, starches, and other materials. Disintegrants may be added as desired.

[0246] "Effective amount" also refers to an amount of a prodrug substance or pharmaceutical composition thereof sufficient to enable or facilitate the amelioration and / or diagnosis of symptoms or signs of a disorder, condition, or pathological condition. The term "effective amount" or "therapeutically effective amount", which is used interchangeably, is defined in some embodiments to mean an amount or quantity of a compound (e.g., ALPHA-1062 or a salt thereof) that is sufficient to induce a significant biological response when administered to a patient. It will be understood that the exact therapeutic dose depends on the age and condition of the patient, the nature of the condition being treated, and is ultimately within the discretion of the attending physician. The choice of dose in this case relates to both the therapeutic effect for the treatment of neurological disorders, as well as the effective amount for antibacterial effect. Both amounts can be evaluated and determined by those skilled in the art.

[0247] The present invention, in some embodiments, encompasses administration of an effective amount of a chemical entity as described herein to a patient or subject in need thereof. By "effective amount" or "therapeutically effective amount" is meant an amount sufficient to ameliorate symptoms or signs of a disorder or physiological condition, or an amount sufficient to allow or facilitate diagnosis of a disorder or physiological condition. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition being treated, the overall health of the patient, the method route and dosage, and the severity of side effects. An effective amount may be the maximum dose or administration protocol that avoids significant side effects or toxic effects. The effect results in an improvement in a diagnostic measure, parameter, or detectable signal of at least 5%, usually at least 10%, more usually at least 20%, most usually at least 30%, preferably at least 40%, more preferably at least 50%, most preferably at least 60%, ideally at least 70%, more ideally at least 80%, and most ideally at least 90%, with 100% being defined as the diagnostic parameter exhibited by a normal subject.

[0248] As used herein, the terms "subject," "patient," or "individual" may be used interchangeably. In preferred embodiments of the present invention, the subject may be an animal, a mammal, or a human, and even more preferably a human.

[0249] As used herein, the term "liquid" refers to its general meaning and includes compositions having a nearly incompressible fluid that conforms to the shape of its container but retains a (nearly) constant volume regardless of pressure.

[0250] As used herein, a "liquid form pharmaceutical composition" is a liquid containing one or more pharma- ceutical active agents suitable for administration to a subject, preferably a mammal, more preferably a human subject. A liquid dosage form is typically a pharmaceutical product with a mixture of drug components and non-drug components (excipients). A liquid dosage form (liquid dosage form) is prepared by (a) dissolving an active drug substance in an aqueous or non-aqueous solvent (e.g., water, glycerin, ether, alcohol), or (b) suspending the drug in a suitable medium, or (c) incorporating the drug substance (drug substance) in an oily or aqueous phase, such as a suspension, emulsion, syrup, or elixir. The solutions described herein are characterized by good solubility in water, as evident for the salts of ALPHA-1062.

[0251] "Emulsions" may also be prepared and adapted for transmucosal administration as disclosed in the art. An "emulsion" is a mixture of two or more liquids that are normally immiscible (unmixable or unblendable). Emulsions are part of a two-phase system of a more general class of substances called colloids. The terms colloid and emulsion are sometimes used interchangeably, but emulsion should be used when both the dispersed and continuous phases are liquids. In an emulsion, one liquid (dispersed phase) is dispersed in another liquid (continuous phase). Examples of emulsions include creams, ointments, liniments (balms), pastes, films, or liquids, depending primarily on their oil-to-water ratio, other additives, and their intended route of administration. Many are topical dosage forms and may be used on the surface of the skin transdermally, transmucosally, ophthalmically, rectally, or vaginally. Highly liquid emulsions may also be used orally, intranasally, or in some cases injected.

[0252] The pharmaceutical composition of the present invention may be manufactured by processes well known in the art, for example, by conventional mixing, dissolving, granulating, pulverizing, emulsifying, encapsulating, encapsulating, dissolving, or lyophilizing processes. The pharmaceutical composition for use according to the present invention may be formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliaries that facilitate the processing of the API into medicament-usable preparations. The appropriate formulation depends on the route of administration selected.

[0253] For injection or transmucosal administration, the API or pharmaceutical composition thereof may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0254] For oral administration, the API, the present invention or pharmaceutical compositions thereof, in some embodiments, can be formulated by combining the API with pharma- ceutically acceptable carriers well known in the art. Such carriers allow a substance, such as the crystalline forms of the present invention, to be formulated as tablets, pills, lozenges, capsules, liquids, gels, syrups, slurries, suspensions, solutions, and the like, for oral ingestion by a patient. Pharmaceutical preparations for oral use can be produced by processing the mixture of granules, optionally by grinding the resulting mixture with solid excipients, after adding other suitable auxiliaries as necessary, to obtain tablets or dragee cores. Useful excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol, cellulose preparations such as corn starch, wheat starch, rice starch, and potato starch, and other materials such as gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP).If desired, disintegrants such as cross-linked polyvinylpyrrolidone, agar, or alginic acid may be added.Salts such as sodium alginate may also be used.

[0255] The compositions of the present invention may also be formulated in some embodiments for parenteral administration, for example, by bolus injection or continuous infusion. Preparations for injection may be presented in unit dosage form, for example, in ampoules or in multi-dose containers, with added preservatives. The compositions may take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory materials such as suspending, stabilizing, and / or dispersing agents.

[0256] In addition to solutions, various liquid compositions comprising ALPHA-1062 or a salt thereof are contemplated, such as emulsions, suspensions, and the like. In one embodiment, the liquid pharmaceutical composition comprising ALPHA-1062 or a salt thereof comprises a sufficient amount or volume (dose) of the composition for multiple administration events to a subject. Thus, the present invention also relates to pharmaceutical compositions comprising ALPHA-1062 or a salt thereof in a solution configured for multiple administration to a subject in need thereof.

[0257] Pharmaceutical compositions for parenteral (any route other than the gastrointestinal tract) administration include aqueous solutions of the water-soluble form of the API. Moreover, suspensions of the drug or prodrug of the present invention, or its pharmaceutical composition, can be prepared in lipophilic vehicles. Suitable lipophilic vehicles include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate and triglycerides, or materials such as liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compound of the present invention or its pharmaceutical composition, to allow for the preparation of highly concentrated solutions.

[0258] In some embodiments, the composition containing active ingredient may be in powder form. For example, the powder may be prepared by constituting it with a suitable vehicle, for example, sterile pyrogen-free water, before use. In some embodiments, the powder formulation is for administration to a subject. For example, the powder formulation, such as the micronized powder formulation, may be administered to a subject by spraying or otherwise applying the powder formulation to the subject, preferably to the mucosal surface of the subject, via intranasal or other methods, such as transmucosal administration.

[0259] The antimicrobial properties of ALPHA-1062 described herein are beneficial not only for liquid formulations such as solutions, suspensions, and emulsions, but also for solid or powder formulations. Powder formulations suitable for transmucosal administration also benefit from the antimicrobial properties of ALPHA-1062 and from the absence of additional antimicrobial preservatives, which is made possible by the knowledge that ALPHA-1062 is itself antimicrobial. Similarly, solid compositions can also benefit from the antimicrobial properties of ALPHA-1062 and the absence of additional antimicrobial preservatives, potentially resulting in reduced microbial contamination and longer storage time for any given ALPHA-1062 composition.

[0260] Determination of a therapeutically effective amount and suitable modes of administration is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. The amount of a composition administered will, of course, depend on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.

[0261] The composition may be presented in a pack or dispenser device, such as an EMA and / or FDA approved kit, which may contain a dosage form containing the active ingredient, if desired. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied by a notice associated with the container in a format prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, which notice reflects the form of the composition or the approval by the agency for human or veterinary administration. Such notice may be, for example, of a label approved by the European Medicines Agency (EMA) and / or the US Food and Drug Administration (FDA) for prescription drugs, or an approved product insert.

[0262] Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0263] A "multi-use dispenser" is known to those skilled in the art and may be provided in the form of any device suitable for multiple administration or application of a compound. In other words, as used herein, it is preferred that the multi-use device does not require opening or refilling between multiple administration events, and is typically stable with sufficient API stability and low microbial load over an extended period of time. In some embodiments, the dispenser may be suitable for administration of any given liquid formulation, preferably a solution, although emulsions and suspensions, and the like, are contemplated.

[0264] "Liquid formulations" are known to those skilled in the art and are common in pediatric or geriatric patient populations. Liquid formulations typically require a stable, dissolved or suspended form of the compound that meets release, bioavailability, and taste / irritation requirements. Both immediate release and sustained release liquid products are contemplated. Liquid formulation strategies typically involve the direct incorporation of the active drug or prodrug in a dissolved or suspended or emulsified state. Alternatives include the incorporation of the active drug or prodrug in the form of a suspended drug-ion exchange resin complex, or in the form of a dissolved or suspended drug-cyclodextrin inclusion complex, or in an emulsified state.

[0265] "Dispensers" configured for transmucosal administration are also known in the art, as are transmucosal administration routes, preferably for oral, nasal, vaginal, and urethral modes. Mucous membranes are relatively permeable and have a rich blood supply, thus allowing rapid uptake of drugs into the systemic circulation and avoiding first-pass metabolism. Oral transmucosal delivery preferably relates to buccal and sublingual routes. This route of drug delivery offers many advantages over other drug delivery approaches, allowing drugs to avoid some of the body's natural "defense mechanisms" such as first-pass metabolism, harsh gastric environment, and potentially intestinal metabolism, due to exposure to the microbial population present in the gut. As an example, several approaches have been used, such as drug delivery via the nasal route by using sprays, pumps, and gels, while mucoadhesive fast-dissolving tablets and solid lozenge formulations are suitable for the oral mucosal route, and vaginal or urethral routes can also be explored using mucoadhesive suppositories, in-situ gels, and foam formulations.

[0266] A preferred dispenser or multi-use administration device according to the present invention relates to any of the multi-use devices mentioned above. In some embodiments, dispensers available from, for example, Nemera (La Verpilliere, France) or Aptar Pharma (IL, USA) are preferred. For example, Nemera offers the Advancia® nasal spray dispenser, which is a high-performance pump with excellent dose consistency and prime retention, anti-clogging actuator, no metal in contact with the formulation, and a hygienic anti-actuation snap-on overcap. As a further example, Aptar Pharma's nasal pump technology eliminates the need for drug manufacturers to add preservatives to nasal spray formulations. Advanced Preservative Free (APF) systems use tip seal and filter technology to prevent contamination of the formulation. A spring-loaded tip seal mechanism is employed with a filter membrane in the vent channel. Notably, the Nemera and Aptar technologies offer numerous advantages such as a metal-free fluid pathway, thereby preventing oxidation of the formulation, providing a preservative-free system and thus an antimicrobial dispenser employing a purely mechanical barrier.

[0267] According to the Advancia (Nemera) technology, air intake into the dispenser is via a venting system with a silicone membrane. This technology and similar technologies have a continuous barrier of homogeneous material that allows air to diffuse through the silicone, acting as a permeable membrane. As a result, the continuous barrier ensures the microbial integrity of the drug. The venting system filters the intake air with a very fine membrane made from a silicone polymer. The silicone membrane is a solid, non-porous material. The membrane is homogeneous and does not contain any holes. The intermolecular distance of the membrane is on the order of nanometers, which allows the passage of air through the membrane, but the silicone membrane structure completely prevents the passage of any liquid or solid particles, including bacteria. The function of the silicone membrane can be compared to an inflated balloon. The balloon is a continuous waterproof material, but gas slowly passes through the walls of the balloon until the internal and external pressures reach equilibrium. In addition, Advancia® PF offers a patented anti-clogging actuator called a closed tip on the top of the system. This mechanism ensures that contaminants cannot enter through the actuator orifice, thus providing protection from crystallization and clogging problems, avoiding evaporation and ensuring good prime retention.

[0268] Additional technologies from Nemera are also contemplated, such as SP270+ and SP370+, which allow very good dose consistency, a wide range of dose volumes (50 μL to 200 μL with different actuators), various neck finishes (screw, snap, and crimp), and are suitable for liquid solutions and suspensions. Alternative technologies from Nemera are contemplated, such as SP27 and SP37, for unregulated markets. Alternatively, continuous valves for nasal and transdermal delivery may be employed, such as Nemera solutions employing pressurized delivery with neutral propellant (nitrogen) or liquid gas, such as CV20 for liquids, valve 6668 for viscous products, or valve 6685 for powders. Additional Nemera dispensing devices are disclosed, for example, in U.S. Pat. No. 9,238,532, which discloses a tip for dispensing liquids for attachment to a container. The valve includes at least two elements that are movable relative to each other, each movable element including a bearing zone for the other movable element. One of the moving elements carries an antimicrobial material on or in the immediate vicinity of a portion of its bearing zone that forms a blocking barrier, and all surfaces of the dispensing tip that come into contact with the interior are free of antimicrobial material. As a further example, US Patent No. 9,345,616 discloses a liquid dispenser device with an air inlet that reliably guarantees the sterility of the contents of the reservoir. The function of taking in air and blocking airborne microorganisms is not performed by an air filter, but by using the gas diffusion properties of certain materials. Therefore, a type of member other than a filter is used, namely a member made of a non-porous polymeric material. Such a member presents the advantage of passing non-contaminated air in a more reliable manner than a filter, which is by definition porous. Other techniques are disclosed in US Patent Application Publication No. 20150043958, US Patent Application Publication No. 8,827,124, US Patent Application Publication No. 8,986,266, and US Patent Application Publication No. 20140231536(A1). All cited patents are incorporated herein by reference in their entirety. By way of further example, Nemera offers advanced preservative-free nasal pumps, a leading technology platform, and a variety of nasal delivery technologies, including VP3 technology, VP6 technology, and VP7 technology.

[0269] As a further example, the nasal spray technology from Aptar offers excellent spray performance, is suitable for viscous drug formulations including solutions, suspensions, and gels, is suitable for gamma radiation sterilization, allows for a wide range of dose volumes from 45 μL to 1,000 μL, and offers a wide range of closures, actuators, and accessories. As a further example, the CPS technology from Aptar is a highly versatile spray pump designed for multi-dose delivery of preserved or non-preserved drug formulations for the nasal route. The CPS may be used for a wide range of other applications including intranasal vaccination. Various advantages include a wide range of dose volumes from 50 μL to 140 μL, does not require repriming even if not used regularly, and can be sterilized by irradiation. The CPS system uses anti-clogging tip seal technology to minimize crystallization in high viscosity and high volatility formulations, the proven CPS filter technology avoids the ingress of contaminating air into the container, and has been fully validated and tested for microbiological integrity, as well as the absence of antimicrobial additives within the pump components and the fluid path components that are metal-free. For example, US Patent No. 9,095,864 to Aptar discloses a nose-shaped fluid dispenser unit and spray device, preferably applied to a sprayer incorporating a closure member for closing the dispenser orifice. The device mechanism disclosed herein relates to completely closing the dispenser orifice when the pump is not in use, and the closure member can be brought into direct cooperation with the dispenser orifice by being resiliently moved toward a closed position by the return spring of the pump, thereby preventing any pathogens or bacteria from penetrating the inside of the device between two actuations, thereby significantly minimizing the risk of contaminating the composition dispensed. As another example, US Patent Application Publication No. 20090294347 discloses a dispensing device for liquid media, having a media reservoir for containing the media, a dispensing opening for dispensing the media from the media reservoir, and a pressure-balancing channel opening into the media reservoir and having a microbiologically active filter arrangement inserted therein.All cited patents are incorporated herein by reference in their entirety.

[0270] The composition according to the present invention may be presented in a pack or dispenser device, such as an FDA approved kit, which may contain a dosage form containing the active ingredient, if desired. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied by a notice associated with the container in a format prescribed by a government agency that regulates the manufacture, use, or sale of pharmaceuticals, which notice reflects the form of the composition or the approval by the agency for human or veterinary administration. Such notice may be, for example, that of the label approved by the US Food and Drug Administration for prescription drugs, or that of an approved product insert.

[0271] Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0272] A preferred condition is a neurological and / or cognitive disorder, preferably one disclosed herein, particularly brain injury associated with TBI or other brain trauma or concussion.

[0273] figure The invention is further illustrated by the figures, which are not intended to limit the scope of the invention. [Brief description of the drawings]

[0274] [Figure 1] Location and effects of controlled cortical impact (CCI) on the rat brain.

[0275] [Diagram 2] Functional tests performed on healthy vehicle- and ALPHA-1062-treated rats to assess sensorimotor and cognitive function following TBI.

[0276] [Diagram 3] Results of sensorimotor function testing evaluating paw obstruction events of the right forelimb of rats while walking on mesh.

[0277] [Figure 4] Results of sensorimotor function testing evaluating right hind limb foot obstruction events during walking on mesh in rats.

[0278] [Diagram 5] 1 is a graph depicting the results of the modified neurological severity score (mNSS) scaling.

[0279] [Figure 6] 4 shows the results of an adhesive removal test on the right forelimb.

[0280] [Figure 7] These are the results of the Morris water maze test.

[0281] [Figure 8] Further results from the Morris Water Maze test.

[0282] [Figure 9] These are the results of a new object recognition test.

[0283] [Figure 10] Weights of test animals over the entire study (days 0-35).

[0284] [Figure 11] ALPHA-1062 significantly reduces lesion volume 35 days after TBI.

[0285] [Figure 12] ALPHA-1062 is neuroprotective and preserves hippocampal structure 35 days after TBI.

[0286] [Figure 13] ALPHA-1062 is neuroprotective, significantly reducing neuronal cell loss (NeuN positive cells quantified 35 days after TBI).

[0287] [Figure 14] ALPHA-1062 significantly enhances dcx+ neuroblasts in the dentate gyrus of the hippocampus (35 days after the TBI event).

[0288] [Figure 15] ALPHA-1062 enhances neurogenesis in the dentate gyrus of the hippocampus - IHC (35 days after TBI).

[0289] [Figure 16] ALPHA-1062 enhances neurogenesis in the dentate gyrus of the hippocampus - cell number (35 days after TBI).

[0290] [Figure 17] ALPHA-1062 significantly reduces p-Tau protein (35 days after TBI).

[0291] [Figure 18] ALPHA-1062 does not alter total Tau protein (35 days after TBI).

[0292] Detailed explanation of the diagram: Figure 1: The diagram illustrates the location and effect of controlled cortical impact (CCI) on the brain of a rat, alive but anesthetized during the intervention. This experiment serves as a simulation of TBI and its effects on the brain. (A) Anatomical scheme (red circle) illustrating the rat skull structure and injury induced in the left parietal cortex to simulate moderate TBI in rats. (B) The distribution of distortion caused by the impact of an object on the rat brain is depicted at the penumbra level. (C) Pathological overview of the injury caused by TBI on the rat brain. (D) The top photograph shows a healthy rat brain (sham operation). The bottom photograph illustrates the size and location of the injury induced in the left parietal cortex of the rat brain to simulate TBI, the rat was sacrificed 7 days after TBI. (E) Hematoxylin and eosin staining of a healthy rat brain section (top) and a rat brain section (bottom) 7 days after TBI-simulated injury (controlled cortical impact CCI) was induced.

[0293] Figure 2: The diagram illustrates the functional tests performed on healthy (sham-operated), vehicle-treated, and ALPHA-1062-treated rats to assess sensorimotor and cognitive functions after TBI. Sensorimotor function tests. (a) The diagram depicts the test to assess the modified neurological severity score (mNSS). (B) The diagram shows the test used to assess the "foot fault" of the animals. The animals have to balance across a grid and the frequency of steps that miss the grid is counted. This test evaluates the functionality of the injured parietal cortex of the rat's brain. (c) The adhesive removal test consists of applying adhesive tape to each of the animal's forepaws and measuring their latency to contact and to removal. This behavior requires correct foot and mouth sensitivity (latency to contact) as well as correct dexterity (latency to removal). Cognitive function tests: (a) The image illustrates the Morris water maze, in which rats are placed in a large circular pool (left picture) and are required to find an invisible platform that allows them to escape from the water by using various visuo-spatial cues. This test is used to study spatial learning and memory and to evaluate hippocampal and select cortical brain area functions. Typically, the rat's path through the pool is traced (right picture), and the time spent near the platform and the time required for the platform to be found by the rat are measured. (b) For the new / novel object recognition (NOR) test, after a habituation period in the test space (leftmost picture), the rat is given the opportunity to explore two identical objects (A) for a given period of time (middle picture). After a delay, the animal is then presented with two objects to explore, one of which is the same as in the first exploration trial (A) and the other is a new object (B) (right picture).Depending on the length of the delay between the two trials, or the extent of the cortical injury, rats will either explore the novel object for a longer period of time, indicating memory for the familiar object, or explore the novel familiar object for the same amount of time, indicating a lack of recollection or loss of memory for the familiar object presented during the first trial.

[0294] Figure 3: The graph shows the results of sensorimotor function testing, which assessed the misstep events of the right forelimb paw of rats walking on mesh. The test was performed as described in Example 1, and rats received either no treatment, vehicle treatment, or ALPHA-1062 treatment without TBI and after a TBI event. The graph shows that animals treated with ALPHA-1062 after undergoing TBI showed significantly fewer paw defects than animals treated with vehicle after TBI. This indicates a significant acute preservation of recovery of locomotor ability, as well as a sustained improvement in speed and extent, in rats treated with ALPHA-1062 after a TBI event, compared to rats only treated with vehicle after TBI. The left bar represents healthy sham rats that did not undergo TBI. The middle bar represents rats that underwent TBI and were subsequently treated with vehicle control treatment. The right bar represents rats treated with ALPHA-1062 after undergoing TBI. Detailed treatment regimens and experimental settings can also be found in Example 1. The sham group contained 8 rats, and the vehicle and ALPHA-1062 treatment groups contained 9 rats each.

[0295] Figure 4: The graph shows the results of sensorimotor function testing, which assessed foot failure events of the right hind paw of rats walking on mesh. The test was performed as described in Example 1, and rats received either sham surgery and no treatment, vehicle treatment, or ALPHA-1062 treatment after a TBI event. The graph shows that animals treated with ALPHA-1062 after undergoing TBI showed significantly less foot failure than animals treated with vehicle after TBI. This indicates a significant acute preservation of locomotor ability, as well as a sustained improvement in speed and extent of locomotor ability, in rats treated with ALPHA-1062 after a TBI event, compared to rats receiving vehicle treatment only. The left bar represents healthy sham rats that did not undergo TBI. The middle bar represents rats that underwent TBI and were subsequently treated with vehicle control treatment. The right bar represents rats treated with ALPHA-1062 after undergoing TBI. Detailed treatment regimens and experimental settings can also be found in Example 1. The sham group contained 8 rats, and the vehicle and ALPHA-1062 treatment groups contained 9 rats each.

[0296] Figure 5: The graph shows the results of the sensorimotor test and depicts the results of the modified neurological severity score (mNSS) scale. The test was performed as described in Example 1, and rats received either no treatment, vehicle treatment, or ALPHA-1062 treatment without TBI and after the TBI event. From day 14 (D14) onwards, ALPHA-1062-treated animals performed statistically significantly better than vehicle-treated animals. The left bar represents healthy sham rats that did not undergo TBI. The middle bar represents rats that underwent TBI and were then treated with vehicle control treatment. The right bar represents rats that underwent TBI and were then treated with ALPHA-1062. The detailed treatment regimen and experimental settings can also be found in Example 1. The sham group contained 8 rats, and the vehicle and ALPHA-1062-treated groups each contained 9 rats.

[0297] Figure 6: The graph shows the results of adhesive removal test of the right forelimb. The test was performed as described in Example 1, and rats received either no treatment, vehicle treatment, or ALPHA-1062 treatment without TBI and after a TBI event. The graph shows that animals that received ALPHA-1062 after a TBI event performed significantly better than vehicle-treated animals from day 7 (D7). The left bar represents healthy sham rats that did not undergo TBI. The middle bar represents rats that underwent TBI and were then treated with a vehicle control treatment. The right bar represents rats that underwent TBI and were then treated with ALPHA-1062. The detailed treatment regimen and experimental settings can also be found in Example 1. The sham group contained 8 rats, and the vehicle and ALPHA-1062 treatment groups each contained 9 rats.

[0298] Figure 7: The graph shows the results of the Morris water maze test. The test was performed as described in Example 1, and rats received either no treatment, vehicle treatment, or ALPHA-1062 treatment without TBI and after the TBI event. The graph shows that from day 33 (D33, first day of evaluation), animals that received ALPHA-1062 after TBI performed significantly better than vehicle-treated animals from that point on. From day 34, ALPHA-1062-treated rats found the underwater platform as did healthy (sham-operated) animals. The left bar represents healthy sham rats that did not undergo TBI. The middle bar represents rats that underwent TBI and were then treated with vehicle control treatment. The right bar represents rats that underwent TBI and were then treated with ALPHA-1062. The detailed treatment regimen and experimental setup can also be found in Example 1. The sham group contained 8 rats, and the vehicle and ALPHA-1062 treatment groups contained 9 rats each.

[0299] Figure 8: The graph shows the results of the Morris water maze test, measuring the time spent in the correct quadrant (green line) of the pool containing the platform. The test was performed as described in Example 1, and rats received either no treatment, vehicle treatment, or ALPHA-1062 treatment without TBI and after the TBI event. The graph shows that animals receiving ALPHA-1062 after TBI performed significantly better than vehicle-treated animals from day 33 (D33) onwards. From day 33 onwards, ALPHA-1062-treated animals spent time in the correct quadrant similar to healthy (sham) animals. Both groups performed statistically significantly better than vehicle-treated animals, which did not show any preference for the correct quadrant. The left bar represents healthy sham rats that did not undergo TBI. The middle bar represents rats that underwent TBI and were subsequently treated with vehicle control treatment. The right bar represents rats that underwent TBI and were treated with ALPHA-1062. Detailed treatment regimens and experimental settings can also be found in Example 1. The sham group contained 8 rats, and the vehicle and ALPHA-1062 treatment groups contained 9 rats each.

[0300] Figure 9: The graph (top panel) shows the results of the novel object recognition test. A schematic of the test is depicted at the bottom of the figure. Briefly, after habituation to the test room, animals were faced with two identical objects (A; left bar, and A'; center left bar), which the animals considered "familiar". The same animals were later exposed to two objects (A = same as before; right bar, and B'; center left bar). * = novel / different object; yellow bars). Rats are naturally exploratory and, when healthy, will explore novel objects (B *) spent more time exploring the novel object. Cognitively impaired animals failed to demonstrate this preference. Both ALPHA-1062-treated animals (right bar of graph) and healthy animals (sham operation; left bar of graph) demonstrated a statistically significant preference for the novel object. In contrast, TBI animals that received only vehicle treatment (center bar of graph) failed to demonstrate any preference for the novel object. The detailed treatment regimen and experimental settings can also be found in Example 1. The sham group contained 8 rats, and the vehicle and ALPHA-1062-treated groups contained 9 rats each.

[0301] Figure 10: Graph shows weight of test animals throughout the study (days 0-35). There was no significant difference in weight between the three treatment groups at each time point during the study (p>0.05). The group of untreated animals (sham operation) shows a significant weight gain from days 20-35 relative to day 0 (intragroup p<0.05). The group of TBI vehicle-treated animals shows a significant weight loss from days 1-6 compared to day 0, and a significant weight gain from days 28-35 compared to day 0 (intragroup p<0.05). The group of ALPHA-1062-treated animals shows a significant weight loss from days 1-7 compared to day 0, and a significant weight gain from days 29-35 compared to day 0 (intragroup p<0.05). Detailed treatment regimens and experimental settings can also be found in Example 1. The sham group contained 8 rats, and the vehicle and ALPHA-1062-treated groups contained 9 rats each.

[0302] Figure 11: ALPHA-1062 treatment significantly reduces lesion volume 35 days after TBI. Panel A: Coronal sections of rat brains from healthy sham-operated animals (left image) or subjected to TBI and vehicle treatment (middle image) or ALPHA-1062 treatment (right image). TBI-related lesions are indicated by circles and the scale bar represents a length of 2 mm. Panel B: Graph depicts lesion volume in percent of brain volume. ALPHA-1062-treated animals (right bar) showed a significant (p<0.05) reduction in lesion volume compared to vehicle-treated mice (middle bar) at 35 days after TBI.

[0303] Figure 12: ALPHA-1062 is neuroprotective and preserves hippocampal structure 35 days after TBI. The hippocampal region is indicated by a circle. To demonstrate that ALPHA-1062 allowed preservation of the hippocampus on the injured side of the brain, the same coronal section shown in Figure 11A is depicted at a higher magnification (circle). The scale bar represents a length of 2 mm.

[0304] FIG. 13: ALPHA-1062 is neuroprotective and significantly reduces neuronal cell loss 35 days after TBI. Results are shown for tissues treated with anti-NeuN immunohistochemistry (IHC) to quantify neuronal cell number. NeuN is also known as the mRNA splicing regulator Fox-3 and is found primarily in the nuclei of mature neurons. Panel A: Coronal images of brain regions by IHC for NeuN protein (Fox-3) 35 days after TBI or in healthy animals (sham operation). Images depict hippocampal region DG, hippocampal region CA1, hippocampal region CA3, and cortex (left to right columns, respectively) of either sham animals (healthy, untreated) or rats with TBI and treated with either vehicle or ALPHA-1062 (top to bottom rows, respectively). Panel B: NeuN positive cells / mm for each of the brain regions. 2 Quantification of neuronal cell counts in the sham-operated animals (left bar) after TBI is depicted. Animals treated with ALPHA-1062 (right bar) showed comparable neuronal cell counts to healthy sham-operated animals (left bar), whereas vehicle-treated animals (middle bar) showed significantly higher neuronal cell counts than sham animals (#) or ALPHA-1062-treated animals (#). * ) suffered significant (p<0.05) neuronal cell loss after TBI compared with either the control or non-control rats.

[0305] Figure 14: ALPHA 1062 significantly enhances DCX+ (doublecortin positive) neuroblasts in the dentate gyrus of the hippocampus (35 days after TBI). Doublecortin (DCX) is an endogenous marker of adult neurogenesis. Panel A: The figure shows iIHC of dentate gyrus sections from untreated sham-operated animals (left image) or animals 35 days after TBI treated with either vehicle (middle image) or ALPHA-1062 (right image). ALPHA-1062-treated animals showed a higher number of DCX-positive stained cells compared to vehicle-treated animals, indicating increased cellular development. ALPHA-1062 treatment resulted in a number of DCX-positive cells comparable to those observed in sham-operated animals. Panel B: Quantification of DCX positive cells per square millimeter of the DG from either sham-operated animals or from animals 35 days after TBI treated with either vehicle (middle bar) or ALPHA-1062 (right bar). * A significantly higher number of DCX-positive cells were present in ALPHA-1062-treated animals compared to healthy sham-operated animals (#), and a significantly reduced number of DCX-positive cells were present in vehicle-treated animals compared to healthy sham-operated animals (#).

[0306] Figure 15: ALPHA-1062 enhances neurogenesis in the dentate gyrus of the hippocampus (35 days after TBI). Panel A: Depicts representative images of NeuN and BrdU double-labeled IHC detection to detect mitotic cells (dark grey arrows) in tissue samples from the dentate gyrus of untreated sham-operated animals or animals treated with either vehicle or ALPHA-1062 for 35 days after TBI (left to right, top panels). ALPHA-1062-treated animals (right image) show a higher number of NeuN and BrdU (double) positive neurons (light grey arrows, merge) compared to vehicle-treated animals (middle image). Panel B: The bottom panel depicts the DG from an ALPHA-1062-treated animal 35 days post-TBI with double labeling of mature granule cell neurons for both NeuN and BrdU (merge; light grey arrows), with the BrdU labeling indicating that the cells were generated from a circulating stem cell population in the days following TBI.

[0307] Figure 16: ALPHA 1062 treatment after TBI enhances neurogenesis in the hippocampal dentate gyrus Quantitation of double positive IHC for NeuN and BrdU: untreated sham-operated animals (left bar), or animals treated with either vehicle (middle bar) or ALPHA-1062 (right bar) 35 days after TBI (for the experiment shown in Figure 15). The number of new granule cell neurons was significantly higher than in the control animals compared to the control animals ( * The neurogenesis was significantly increased in animals treated with ALPHA-1062 compared to both the sham-operated (#) and healthy animals (p<0.05). This observation indicates an increase in neurogenesis, which leads to improved tissue recovery after TBI when ALPHA-1062 treatment is administered.

[0308] Figure 17: ALPHA 1062 treatment significantly reduces pathological p-Tau accumulation (35 days after TBI). The top panel shows brain sections of sham-operated, ALPHA-1062, or vehicle-treated animals labeled with an antibody against p-Tau. p-Tau immunoreactive areas, including the cortex (CT), dentate gyrus (DG), CA3, and CA1 regions of the hippocampus in the ipsilateral hemisphere, were imaged and quantified by Image J software (NIH). The bottom panel shows quantification of IHC intensity. Sham-operated animals (left bar) show relatively low levels of p-Tau, while vehicle (control)-treated TBI animals (middle bar) show high levels of p-Tau. TBI animals treated with ALPHA-1062 (right bar) show a significant reduction in p-Tau levels compared to vehicle controls. The number of animals in each group is shown.

[0309] Figure 18: ALPHA 1062 treatment does not alter total Tau protein (35 days after TBI). The top panel shows brain sections of untreated sham-operated, ALPHA-1062-treated, or vehicle-treated animals labeled with an antibody against total Tau. Tau immunoreactivity in the cortex (CT), dentate gyrus (DG), CA3, and CA1 regions of the hippocampus of the ipsilateral hemisphere was imaged and quantified by Image J software (NIH). The bottom panel shows the quantification of the resulting signal. No significant differences in the levels of total Tau were observed between any of the animal groups. The number of animals in each group is shown. EXAMPLES

[0310] The present invention is further illustrated by the following examples, which are intended to further illustrate the present invention as working examples and do not represent a limiting description of the present invention.

[0311] Example 1: Evaluation of the efficacy of ALPHA-1062 in rats following traumatic brain injury In this example, the efficacy and use of the substances and pharmaceutical compositions according to one embodiment of the present invention was carried out in an exemplary 35-day study in a controlled cortical impact injury model.The purpose of this study was to determine the efficacy of ALPHA-1062 (galantamine benzoate gluconate salt) on functional and histological outcomes in young adult male rats after moderate traumatic brain injury (TBI) induced by controlled cortical impact (CCI).

[0312] summary: Compared to vehicle (purified water) treated controls, ALPHA-1062 treatment was initiated 2 hours after injury (4.5 mg / kg IN) twice daily (6 hour intervals) for 35 days: (1) Significant improvement in foot injury, adhesive removal, and motor-sensory function recovery as measured by the mNSS test.

[0313] (2) Significant improvement in cognitive function recovery as measured by the NOR and MWM tests 5 weeks after injury.

[0314] (3) did not significantly change body weight over the 35-day study (no evidence of overt toxicity).

[0315] (4) A significant reduction in lesion size measured 35 days after injury.

[0316] (5) A significant reduction in neuronal cell loss in the ipsilateral cortex and hippocampus.

[0317] (6) Significantly enhanced neurogenesis, including an increase in DCX+ neuroblasts and BrdU / NeuN+ new neurons in the DG.

[0318] (7) The significant reduction in pathological p-Tau accumulation in the injured cortex and hippocampus 35 days after injury suggests that ALPHA-1062 reduces neurodegeneration in rats after moderate TBI induced by CCI.

[0319] Acute IN administration (2 hours after injury) of ALPHA 1062 twice daily treatment provides neuroprotection, reduces pathological p-Tau accumulation, stimulates neurogenesis, and significantly improves sensorimotor and cognitive recovery in a rat contusion model of TBI compared with vehicle treatment. These data suggest that ALPHA 1062 has potential as a treatment for TBI.

[0320] method: Young adult (2-3 months) male Wistar rats were subjected to moderate traumatic brain injury (TBI) induced by controlled cortical impact (CCI). Animal models of TBI, including the CCI model, and functional testing have been described in detail in peer-reviewed publications (Xiong et al., 2013, Nat Rev Neurosci.; Zhang et al., 2016, J Neurosurg.; Zhang et al., 2016, Neurochem Int.; Zhang et al., 2021, J Neurotrauma). The injury consisted of a single impact of 4.0 m / s, resulting in a 2.5 mm deformation to the left parietal cortex. Additionally, a group of animals was left healthy / uninjured to serve as healthy controls.

[0321] The animals were divided into three groups: (1) TBI + vehicle (purified water, n = 10) (2)TBI+ALPHA-1062(4.5mg / kg)(n=10) (3) Healthy / no injury and no treatment (sham surgery) (n=10)

[0322] To obtain 8 rats per study group, taking into account a 20% mortality rate, 10 rats (8 / 0.8=10) were included. The total number of male rats used as planned was 30.

[0323] The substance used in this study was ALPHA-1062 at 4.5mg / kg. Purified water was administered intranasally (IN) as a control treatment (vehicle). ALPHA-1062 and control treatments were started 2 hours after injury and continued twice daily (6 hours apart) for 35 consecutive days.

[0324] In addition to ALPHA-1062 or vehicle control treatment, 1 day after injury, Bromodeoxyuridine (BrdU) was administered by intraperitoneal injection at 100 mg / kg for 7 consecutive days to allow for monitoring of cell proliferation. Bromodeoxyuridine (BrdU) is a thymidine analogue that is incorporated into the DNA of dividing cells during the S phase of the cell cycle.

[0325] The following statistical analysis was applied to assess data for normality: data transformation was considered if data were abnormal. Data were analyzed using GraphPad Prism Software 9.0. All data herein are presented as mean ± standard deviation (SD) and were analyzed by one-way analysis of variance (ANOVA) or two-way analysis of variance (treatment × time) followed by post-hoc Tukey's multiple comparison test (more than two groups). Differences between means were considered statistically significant when p was <0.05.

[0326] During the in vivo portion of the study, cognitive function testing was performed by performing the Morris water maze (MWM) test and novel object recognition test on all animals 5 weeks after TBI, with the MWM test being performed on days 31-35 after TBI (see Table 1).

[0327] In addition, neurological function testing was performed by performing the foot injury test and observation to determine the modified neurological severity score (mNSS) on days 1, 7, 14, 21, 28, and 35 after TBI (see Table 1). The mNSS is a composite score that evaluates motor, sensory, balance, and reflex function. Details of the tests and scoring system can be found in Table 2.

[0328] [Table 2]

[0329] The Morris Water Maze (MWM) is a test used in behavioral neuroscience to study spatial learning and memory, for example in rats. It allows studying spatial learning and memory and can also be used to evaluate damage to the hippocampus and select cortical areas of the brain. The MWM can be used to evaluate the effects of lesions on the brain in areas involved in memory, cognitive function, and spatial learning. In the Morris Water Maze test, for example, rats are placed in a circular pool and are required to find an invisible underwater platform that allows them to rest and exit the water. Typically, rats are tracked while swimming in the pool and parameters such as the time spent swimming in the quadrant of the pool containing the platform or in close proximity to the platform, and the total time spent to find the platform (escape latency) are measured.

[0330] In the adhesive removal test, adhesive tape is applied to the right forepaw of the animals and both the latency to contact the adhesive (sensory perception) and the latency to remove the adhesive (locomotor activity) are assessed by this test.

[0331] The rat novel / new object recognition (NOR) assay is a relatively high-throughput, robust and sensitive measure for evaluating compounds for cognition-enhancing activity. For testing, rats are given the opportunity to explore two identical objects for a given period of time. After a delay, the animals are then presented with two objects to explore, one of which is the same as in the first exploration trial and the other is a new object. Depending on the functionality of the brain, rats will either explore the novel object for a longer period of time, indicating memory for the familiar object, or explore the novel familiar object for the same time, indicating lack of recall or loss of memory for the familiar object presented during the first trial.

[0332] The modified neurological severity score (mNSS) is a rating scale of neurological functionality and includes a composite of motor (muscle status and abnormal movements), sensory (visual, tactile, and proprioceptive), reflex, and balance tests. This test is used to evaluate the grade of neurological impairment and neurological damage in terms of aspects of movement, overground walking, sensation, motor coordination, reflex, and abnormal movement. It is usually performed with rodents, such as rats, and the examiner is blinded to the treatment group. Four aspects are observed, including abnormal movements or lack of reflexes, beam balance test, sensory function, and locomotor function. The baseline for normal animals is 0 points.

[0333] * Inability to perform a test, absence of a tested reflex, or abnormal movement was given a score value of 1.

[0334] [Table 3]

[0335] Histological evaluation was performed including a quantitative image analysis protocol; followed by statistical analysis of the data. Histology: All rats were sacrificed after the final Morris Water Maze (MWM) test 35 days after TBI for the following histological analyses: lesion volume (H&E), neuronal cell loss in the injured cortex and hippocampus (NeuN), doublecortin (DCX, neuroblasts), NeuN / BrdU (newborn neurons), pathological p-Tau (AT8), and total Tau IHC.

[0336] For tissue preparation, rats were anesthetized and perfused transcardially with saline followed by 4% paraformaldehyde in 0.1 M PBS, pH 7.4. Rat brains were removed and immersed in 4% paraformaldehyde for 2-4 days. Using a rat brain matrix (Activational Systems Inc.), each forebrain was cut into 2 mm thick coronal blocks for a total of 7 blocks per animal, from bregma 5.2 mm to bregma -8.8 mm. Tissues were embedded in paraffin and a series of 6 μm thick slides were cut.

[0337] For immunohistochemistry (IHC), antigen retrieval was performed by boiling the sections in 10 mM citrate buffer (pH 6.0) for 10 min. After washing with PBS, the sections were incubated with 0.3% H2O2 in PBS for 10 min, blocked with 1% BSA containing 0.3% Triton®-X100 for 1 h at room temperature, and incubated with mouse anti-doublecortin antibody (1:200; DCX, Santa Cruz Biotechnology, Santa Cruz, CA) and monoclonal mouse anti-NeuN antibody (1:400; Chemicon, Millipore) at 4°C overnight. For negative controls, the primary antibody was omitted. After washing, the sections were incubated with biotinylated anti-mouse antibody (1:200; Vector Laboratories, Inc.) for 30 min at room temperature. After additional washing, sections were incubated with avidin-biotin-peroxidase system (ABC kit, Vector Laboratories, Inc.), visualized with diaminobenzidine (Sigma), and counterstained with hematoxylin (results shown, for example, in Figures 13 and 14).

[0338] For immunofluorescence staining (e.g., results shown in Figures 15 and 16), brain sections were boiled for 10 min in 10 mM citrate buffer (pH 6) after deparaffinization and rehydration. After washing with PBS, sections were incubated in 2.4 N HCl at 37°C for 20 min. Sections were incubated with 1% BSA containing 0.3% Triton®-X-100 in PBS. Sections were then incubated overnight at 4°C with mouse antibody against BrdU (1:200). For negative controls, the primary antibody was omitted. Cy3-conjugated anti-mouse antibody (1:400; Jackson ImmunoResearch, West Grove, PA) was added to the sections for 2 h at room temperature. Each step was followed by 3 rinses in PBS for 5 min. Sections were then incubated with different rabbit antibodies against NeuN (1:200) and, after washing, incubated with FITC-conjugated goat anti-rabbit antibody for 2 h at room temperature. Tissue sections were mounted with Vectashield mounting medium (Vector Laboratories, Burlingame, CA).

[0339] Immunofluorescence staining of Tau and p-Tau (results shown in Figure 17 and Figure 18) was performed as follows: After deparaffinization and rehydration, brain sections were boiled in 10 mM citrate buffer (pH 6) for 10 min. After washing with PBS, sections were incubated with 1% BSA in PBS. Sections were then incubated with chicken antibody against total TAU (1:100, average) or mouse anti-p-Tau (1:500, Ser202 / Thr205 Fisher, MN1020, AT8) overnight at 4°C. After washing, sections were incubated with FITC-conjugated goat anti-chicken or mouse secondary antibody (1:400, Jackson ImmunoResearch Inc) for 2 h at room temperature. After washing, sections were stained with DAPI (1:10,000) for 2 min and rinsed with water for 5 min. Tissue sections were mounted with Vectashield mounting medium (Vector laboratories, Burlingame, CA).

[0340] For cell counting and quantification, NeuN+ cells were surveyed in the ipsilateral cortex, DG, CA1, and CA3 regions of the ipsilateral hippocampus. For the analysis of neurogenesis, neuroblasts were defined by DCX+ cells. The number of DCX+ cells was surveyed in the granule cell layer of the DG of the ipsilateral hippocampus. Newly generated neurons were identified by cells with colocalization of NeuN and BrdU immunoreactivity. We focused on the DG and its subregions, including the subgranular zone, granule cell layer, and molecular layer. The number of BrdU+ cells (red fluorescent) and NeuN (green fluorescent) / BrdU co-labeled cells (yellow after merge) were counted. Fields of interest were digitized under a light microscope (Nikon, Eclipse 80i) at either 200 or 400 magnification using a CoolSNAP color camera (Photometrics) interfaced with a MetaMorph image analysis system (Molecular Devices). Immunopositive cells were determined, divided by the measured area, and presented as number per square mm. Cell counts were performed by an observer blinded to the individual treatment status of the animals. Tau or p-Tau immunoreactive areas, including the cortex, dentate gyrus, CA3 region, and CA1 region of the ipsilateral hemisphere, were imaged and quantified by Image J software (NIH).

[0341] For statistical analysis, all data are presented as mean ± standard deviation (SD) and assessed for normality. If data are abnormal, data transformation is considered. Data were analyzed by one-way analysis of variance (ANOVA) or two-way analysis of variance (treatment × time) followed by post-hoc Tukey's multiple comparison test (more than two groups) using GraphPad Prism Software 9.0. Differences between means were considered statistically significant when p was <0.05.

[0342] result ALPHA-1062 treatment acutely preserves and sustains improvement in locomotor recovery in forelimb and hindlimb paw injury tests The animals were monitored as they walked across the mesh for the number of times the right forelimb "stepped off" the appropriate position, indicating a decrease in locomotor activity, indicative of a left-sided brain injury. The results are depicted in Figure 3. Tests were also performed on the right hindlimb (Figure 4).

[0343] ALPHA-1062 treatment (Figures 3 and 4, grey bars) resulted in statistically significant preservation of locomotor performance from day 1 of treatment and sustained, statistically significant, improved recovery throughout the study compared to vehicle-treated animals (Figures 3 and 4, center bars). By day 28 (D28), the performance of ALPHA-1062-treated animals was statistically indistinguishable from that of non-injured (Sham) animals (Figures 3 and 4, right bars vs. left bars). The acute preservation of locomotor performance suggests that early administration of ALPHA-1062 after TBI likely reduced the extent of brain damage. Vehicle-treated animals (Figures 3 and 4, center bars) show an inherent level of functional recovery after TBI.

[0344] ALPHA-1062 treatment improves outcomes in modified neurological severity score (mNSS) The mNSS is a composite score that assesses motor, sensory, balance, and reflex function (details of the test and scoring system can be found in Table 2). From day 14 (D14) of recovery onwards, ALPHA-1062-treated animals performed statistically significantly better (Figure 5, right bar) than vehicle-treated animals (Figure 5, middle bar), and this effect persisted throughout the study period compared to vehicle-treated animals.

[0345] ALPHA-1062 treatment improves motor performance in the right forepaw adhesive removal test The results of the adhesive removal test are shown in Figure 6. Starting on day 7 of recovery (D7) and throughout the remainder of the study, ALPHA-1062-treated animals (Figure 6, right bars) performed this motor ability similarly to uninjured (Sham) animals (Figure 6, left bars).

[0346] ALPHA-1062 administration improves spatial learning and memory in the MWM test The effects of ALPHA-1062 treatment after TBI were assessed using the Morris Water Maze (MWM). The Morris Water Maze is a test of rodent spatial learning that relies on distal cues to locate an underwater escape platform from a starting point around an open swimming area. Spatial learning is assessed over repeated trials, and reference memory is determined preferentially for the platform area (the "correct quadrant") in the absence of the platform. Figure 7 depicts the results of the final testing day of the MWM test, where rats swam in a pool of water and were assessed for latency to find the platform (time from start to goal = platform). On days 33-35 (D33-D35) of testing, ALPHA-1062-treated animals (Figure 7, right bars) were able to find the hidden escape platform as well as uninjured (Sham) animals (Figure 7, left bars). Both groups performed statistically significantly better than vehicle-treated animals (middle bar).

[0347] In addition, the percentage of time (% time) that the rats spent in the "correct" quadrant of the pool, i.e., the quadrant where the platform was submerged in the previous session, was also assessed. The results can be seen in Figure 8. On days 33-35 of testing, ALPHA-1062-treated animals (Figure 8, right bars) spent comparable time in the correct quadrant (encircled in green) as non-injured (sham-operated) animals (Figure 8, left bars). Both groups performed statistically significantly better than vehicle-treated animals (Figure 8, center bars), which failed to show any preference for the correct quadrant throughout the entire test.

[0348] ALPHA-1062 treatment enables cognitive recovery after TBI To evaluate the recovery of cognitive function after TBI, an object recognition test was performed. The test workflow is depicted in the bottom of Figure 9, and the results, i.e., the percentage of time spent exploring each object, are depicted in the top of Figure 9. The animals were placed in a room with two identical objects (A (left bar in the top graph of Figure 9) and A' (middle bar)), where the animals became "familiar" with these objects. The same animals were later exposed to two objects (A = same as before (right bar) and B = novel / different object (yellow bar)). Rats are naturally exploratory and, in healthy cases, spent more time exploring the novel object (B), which is represented by a higher value of B in the graph of Figure 9 (right bar shown in the top graph of Figure 9). (A). Cognitively impaired animals fail to show this preference since they do not remember the familiar object. Thus, cognitive impairment can be assumed when rats spend equal time exploring the familiar and novel objects. Both ALPHA-1062-treated animals (Figure 9, right bars in top graph) and unlesioned animals (Figure 9, left bars) showed a statistically significant preference for the novel object (B). In contrast, lesioned animals that received only vehicle treatment (Figure 9, middle bars) failed to show any preference for the novel object.

[0349] [Table 4]

[0350] ALPHA-1062 administration did not alter body weight or show any obvious toxicity As can be derived from FIG. 10, rats treated with ALPHA-1062 (FIG. 10, right bar) did not undergo significant changes in body weight compared to vehicle-treated or untreated animals (FIG. 10, center bar and left bar; no significant differences (P>0.05) in body weight were detected among the three groups at each time point). Vehicle- and ALPHA-1062-treated animals showed significant weight loss during the first week of treatment only when compared to day 0 of their own treatment group. Vehicle-treated animals showed significant weight loss (p<0.05) on days 1-6 (D1-6) compared to day 0 (D0), but showed significant weight gain (p<0.05) on days 28-35 (D28-35) compared to day 0. ALPHA-1062-treated animals showed a significant (p<0.05) weight loss on days 1–7 (D1–7) compared to day 0 (D0), but showed a significant weight gain from days 29–35 (D29–35) compared to day 0. Untreated (sham) animals showed a significant (P<0.05) weight gain on days 20–35 (D20–35) compared to day 0 (D0) compared to their weight on day 0 (D0).

[0351] Thus, ALPHA-1062 administration did not result in overt toxicity in treated animals compared to vehicle animals.

[0352] ALPHA-1062 significantly reduces lesion volume 35 days after TBI.

[0353] After sacrifice and quantification of tissue loss 35 days after TBI, ALPHA-1062-treated animals showed a significant (p<0.05) reduction in lesion volume compared to vehicle-treated mice (see also FIG. 11). As shown in detail in FIG. 12, ALPHA-1062 treatment allowed preservation of hippocampal structure on the injured side of the brain. Therefore, it can be concluded that ALPHA-1062 treatment exhibits a neuroprotective effect, resulting in the preservation of brain structure 35 days after TBI.

[0354] ALPHA 1062 is neuroprotective and significantly reduces neuronal cell loss.

[0355] IHC using anti-NeuN antibody to quantify the number of neurons revealed that ALPHA-1062 treatment reduced neuronal loss in brain regions DG, CA1, CA3, and cortex analyzed 35 days after TBI compared to vehicle-treated animals. Importantly, ALPHA-1062 treatment resulted in neuronal cell numbers that were not significantly different from sham-operated animals. However, vehicle-treated animals suffered significant (p<0.05) neuronal cell loss after TBI compared to either sham or ALPHA-1062-treated animals (see also FIG. 13).

[0356] ALPHA 1062 significantly enhances doublecortin-positive neuroblasts in the dentate gyrus of the hippocampus.

[0357] IHC for doublecortin (DCX), an endogenous marker of adult neurogenesis, demonstrated that ALPHA-1062-treated animals showed a higher number of DCX-positive cells compared to samples from vehicle-treated animals, and a comparable number to animals from the sham-operated cohort. Vehicle-treated animals subjected to TBI showed a significantly reduced number of DCX-positive cells compared to sham-operated animals (see also FIG. 14). Thus, it can be concluded that ALPHA-1062-treated animals showed increased cell regeneration when assessed 35 days after TBI.

[0358] ALPHA-1062 enhances neurogenesis in the dentate gyrus of the hippocampus.

[0359] In addition, NeuN and BrdU double-labeled IHC was performed to detect mature and newly born granule neurons. ALPHA-1062-treated animals showed significantly (p<0.05) higher numbers of NeuN and BrdU double-positive granule neurons compared to both vehicle-treated and sham-operated animals. The higher number of IHC double-positive cells observed in ALPHA-1062-treated animals indicates increased neurogenesis in the dentate gyrus compared to vehicle-treated animals (see also FIG. 15), suggesting that ALPHA-1062 treatment increased neuronal recovery and neurogenesis in brain tissue.

[0360] ALPHA 1062 significantly reduces pathological p-Tau accumulation in the injured cortex and hippocampus.

[0361] IHC for total Tau and p-Tau (results shown in Figure 17 and Figure 18) was performed. Tissues from sham-operated, ALPHA-1062-treated, or vehicle-treated animals were labeled with antibodies against p-Tau or Tau. Immunoreactive areas, including the cortex (CT), dentate gyrus (DG), CA3, and CA1 regions of the ipsilateral hemisphere, were imaged and quantified by Image J software (NIH). As shown in Figure 17, sham-operated (left bar) shows relatively low levels of p-Tau, while vehicle (control)-treated TBI animals (middle bar) show high levels of p-Tau. TBI animals treated with ALPHA-1062 (right bar) show a significant decrease in p-Tau levels compared to vehicle controls. Furthermore, as shown in Figure 18, ALPHA 1062 treatment does not alter total Tau protein. This data indicates that ALPHA-1062 treatment reduces pathological p-TAU accumulation after CCI-induced moderate TBI.

[0362] conclusion Compared to vehicle controls, ALPHA-1062 treatment significantly improved motor and sensory function recovery in treated rats after TBI as measured by paw injury, adhesive removal, and mNSS tests.

[0363] Also, 5 weeks after TBI, NOR and MWM tests showed significantly improved cognitive recovery in rats when ALPHA-1062 treatment was administered.ALPHA-1062 treatment did not significantly change body weight over the 35-day study compared to untreated or vehicle-treated animals.Therefore, no obvious evidence of toxicity could be observed in ALPHA-1062-treated rats.

[0364] Further ALPHA-1062 treatment significantly reduced the lesion size measured 35 days after injury and also significantly reduced neuronal cell loss in the ipsilateral cortex and hippocampus. Surprisingly, the degree of neuroprotection afforded by ALPHA-1062 treatment preserved neurons in areas affected by TBI to a degree that was indistinguishable from the numbers determined in sham (non-TBI) control animals.

[0365] ALPHA-1062 treatment also significantly enhanced neurogenesis, including an increase in DCX+ neuroblasts and BrdU / NeuN+ new neurons in the DG.

[0366] In conclusion, acute intranasal (IN) administration after TBI (e.g., 2 hours after injury) followed by regular administration (e.g., daily) of ALPHA-1062 provides neuroprotection, stimulates neurogenesis, and significantly improves sensorimotor and cognitive recovery in a rat contusion model of TBI compared to vehicle treatment.

[0367] ALPHA-1062 also significantly reduced pathological p-Tau accumulation, reducing the potential for further tissue damage driven by p-Tau.

[0368] These data suggest that ALPHA-1062 has potential as a treatment for acute TBI, for example by intranasal administration.

[0369] Example 2: Antibacterial properties of ALPHA-1062 To evaluate the antimicrobial properties of ALPHA-1062, the preservative challenge test (USP 51) from USP Chapter 51 was employed. USP 51 is a common method used to measure the effectiveness of preservatives. Similar to the preservative efficacy screen, it is used to evaluate the effectiveness of preservatives in cosmetics, personal care products, and drug products. Preservatives are antimicrobial ingredients that are typically added to aqueous product formulations to help maintain the safety of the product by inhibiting the growth and reducing the number of microbial contaminants.

[0370] In the context of the present invention, ALPHA-1062 (in the form of gluconate salt) was evaluated instead of (as well as) preservatives. No additional preservatives were added to the ALPHA-1062 gluconate preparations, rather the antimicrobial effect of the drug itself (ALPHA-1062) was evaluated. Thus, ALPHA-1062 is the antimicrobial drug in the assays and experiments described below.

[0371] The USP 51 potency test utilizes five microorganisms (three bacteria and two fungi) for potency testing. Each microorganism is a known strain of a pathogenic microorganism and represents a wide range of microbial physiology.

[0372] Following standard guidelines of USP 51, cultures of the following microorganisms were applied: Candida albicans (ATCC No. 10231), Aspergillus brasiliensis (ATCC No. 16404), Escherichia coli (ATCC No. 8739), Pseudomonas aeruginosa (ATCC No. 9027), and Staphylococcus aureus (ATCC No. 6538).

[0373] The test was carried out in five sterile capped bacteriological containers of suitable size into which sufficient quantities of product were transferred. Each container was inoculated with one of the prepared and standardized inocula and mixed accordingly. The volume of the suspended inoculum used was 0.5%-1.0% of the volume of the product. The concentration of the test microorganism added to the product was determined according to the standard guidelines for category 2 products, with the final concentration of the test preparation after inoculation being 1 × 10 per mL of product. 5 ~1×10 6 The concentration was such that CFU were obtained.

[0374] The initial concentration of viable microorganisms in each test preparation was estimated based on the concentration of microorganisms in each of the standardized inocula determined by plate counting.

[0375] The inoculated vessels were incubated at 22.5±2.5° C. and each vessel was sampled at appropriate intervals as shown in Table 3. Any changes observed in the appearance of the cultures were recorded at these intervals. By using plate count procedures, the number of CFU present in each test preparation was determined for the applicable intervals.

[0376] Inactivators (neutralizers) of specific antimicrobial agents were incorporated during plate counts or in the appropriate dilutions prepared for plating. These conditions were determined in the validation study for that sample based on the media and microbial recovery incubation time conditions listed in Table 2. Using the calculated concentration of CFU / mL present at the start of the test, the log10 change in concentration of CFU / mL for each microorganism was calculated at the applicable test interval, and the changes were expressed as log reductions.

[0377] [Table 5]

[0378] As can be seen from the data provided, the gluconate salt of ALPHA-1062 exhibits strong antimicrobial efficacy against all five organisms tested in USP 51. Category 2 USP criteria were adopted to determine the presence of antimicrobial efficacy. For bacteria, antimicrobial efficacy is evident if at least a 2.0 log reduction from the initial count is evident on day 14 and no increase from the day 14 count is evident on day 28. For yeasts and molds, antimicrobial efficacy is evident if no increase from the initial calculated count is determined on days 14 and 28. Thus, with respect to the above data, efficacy against bacteria and fungi exceeded the requirements for demonstrating antimicrobial efficacy for a Category 2 product.

Claims

1. A pharmaceutical composition for use in treating confirmed or suspected traumatic brain injury (TBI) in a subject, the pharmaceutical composition comprising the compound ALPHA-1062 or a salt thereof.

2. 10. The pharmaceutical composition for use according to claim 1, wherein the composition is in liquid form.

3. The pharmaceutical composition for use according to claim 2, wherein the composition is administered intranasally.

4. 4. The pharmaceutical composition for use according to any one of claims 1 to 3, wherein the composition is self-preserving and antimicrobial, and wherein the composition has no added antimicrobial preservatives.

5. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the composition is present in a multi-use dispenser configured for intranasal administration.

6. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the subject having or suspected of having a TBI exhibits one or more symptoms selected from the group consisting of dizziness, balance problems, headache, nausea, vomiting, light sensitivity, memory impairment, sleep abnormalities, poor concentration, and visual disturbances.

7. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the subject having or suspected of having moderate or severe TBI exhibits one or more symptoms selected from the group consisting of weakness in the arms and legs, problems with balance and coordination, severe or increasingly severe headaches, impaired sensory perception, impaired cognitive abilities, memory impairment, impaired communication and learning, personality changes, behavioral abnormalities, and visual and hearing impairments.

8. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the subject having or suspected of having severe TBI exhibits one or more symptoms selected from the group consisting of paralysis, coma, fainting, dilated pupils, loss of cerebrospinal fluid from the ears or nose, loss of bowel and / or bladder control, breathing problems, slow pulse, breathing problems, slow breathing rate with elevated blood pressure, and ptosis or facial weakness.

9. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the traumatic brain injury (TBI) results in or is accompanied by one or more injuries selected from the group consisting of hematoma, contusion, intracerebral hemorrhage, subarachnoid hemorrhage, diffuse injury, diffuse axonal injury, ischemia, primary brain injury, and secondary brain injury.

10. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the traumatic brain injury (TBI) is caused by a closed head injury or a penetrating head injury.

11. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the traumatic brain injury (TBI) is caused by a head injury resulting from an accident selected from the group consisting of a fall, a motor vehicle related accident, a blow or blow to the head from or against an object, an explosion (causing blast-related TBI), a sports related accident, interpersonal physical violence or violence by other means.

12. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the subject having or suspected of having TBI is an infant under 4 years of age, or a child between 4 and 12 years of age, or an adolescent between 12 and 17 years of age.

13. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the subject having or suspected of having TBI is an adult selected from adults aged 18 to 65 years, or elderly adults over 65 years.

14. 3. A pharmaceutical composition for use according to claim 1 or 2 for stimulating and / or enhancing neurogenesis and / or neuronal recovery in the brain of a subject confirmed or suspected of having a traumatic brain injury (TBI).

15. 3. A pharmaceutical composition for use according to claim 1 or 2 for preventing, inhibiting and / or reducing neuronal cell loss in the brain of a subject confirmed or suspected of having traumatic brain injury (TBI).

16. 3. A pharmaceutical composition for use according to claim 1 or 2 for reducing the size of one or more lesions and / or damage in the brain of a subject confirmed or suspected of having a traumatic brain injury (TBI).

17. 3. A pharmaceutical composition for use according to claim 1 or 2 for preserving brain tissue and / or neuronal tissue in the brain of a subject confirmed or suspected of having a traumatic brain injury (TBI).

18. 3. A pharmaceutical composition for use according to claim 1 or 2 for reducing and / or preventing the level and / or formation of p-Tau in the brain of a subject confirmed or suspected to have had a traumatic brain injury (TBI).

19. 3. A pharmaceutical composition for use according to claim 1 or 2, wherein the traumatic brain injury (TBI) is associated with trauma to the nasal cavity and / or for use in the treatment of traumatic brain injury (TBI) and the combined treatment and / or prevention of microbial infection of the nasal cavity.

20. 3. A pharmaceutical composition for use according to claim 1 or 2, wherein the traumatic brain injury (TBI) is associated with a disruption of the blood-brain barrier and / or for use in the treatment of traumatic brain injury (TBI) and the combined treatment and / or prevention of microbial infections of the central nervous system.

21. The pharmaceutical composition for use according to claim 1 or 2, wherein the compound ALPHA-1062 or a salt thereof is present in a concentration of 1 to 500 mg / mL.

22. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein said compound ALPHA-1062 is present as a gluconate salt in a concentration of 50 to 100 mg / mL.

23. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the composition is administered in a dose of 0.1 to 200 mg.

24. 3. The pharmaceutical composition for use according to claim 1 or 2, wherein the composition is administered within 24 hours of injury.

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