Iodbr1 and derivatives thereof for treating sleep disorders

Abstract

Described are compositions comprising a compound of formula I: in which R represents H, or optionally substituted C1-15 alkyl, C1-15 alkenyl or C1-15 alkynyl; or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, for use in the treatment or prevention of a sleep disorder, together with methods of treating or preventing a sleep disorder in a subject or of treating or preventing a symptom or co-morbidity associated with a sleep disorder.

Description

[0001] TREATMENT OF SLEEP DISORDERS

[0002] Field of the Invention

[0003] This invention relates to compositions comprising idoBRI or related compounds for the treatment or prevention of sleep disorders.

[0004] Background of the Invention

[0005] Iminosuqar acids and idoBRI

[0006] Iminosugar acids (ISAs) constitute a subclass of the more widely distributed class of phytochemicals known as iminosugars. Many known ISAs are present as secondary metabolites in plant tissues (where they may play a role in defence). While iminosugars are widely distributed in plants (Watson etal. (2001) Phytochemistry 56: 265-2951), the iminosugar acids are much less widely distributed.

[0007] The ISA idoBRI, (2R,3R,4R,5S)-3,4,5-trihydroxypiperidine-2-carboxylic acid, also referred to as 6-epiBR1, 2R,3R,4R,5S-Trihydroxypipecolic acid and ido-BR1 in the literature, has the formula:

[0008] "

[0009]

[0010] It has been shown to be anti-inflammatory and reduces various inflammatory signals such as TNF-alpha both in vivo and in vitro (Nash et al., 2020, ACS Omega ao-2020-02092f; WO2013 / 054070; PCT / GB2012 / 000768). IdoBRI has also been shown to inhibit inflammatory signals produced by brain microglia in vitro (Olajide etal., 2022, Molecules.

[0011] 27, 3342) and thought to be involved in neurodegeneration. The anti-inflammatory activity of cucumber extract appears to be solely due to the idoBRI content (WO2013 / 054070). IdoBRI is also inhibitory to sialidases and has activity on microbial growth (W02021 / 019194). The possible mechanism of idoBRI in osteo-arthritis has been studied and seems to involve, at least in part, interaction with a hyaluronic acid receptor CD44 which has increased expression in osteo-arthritis (Nash et al., 2020, ACS Omega ao-2020-020921). IdoBRI was also shown to reduce activation of the transcription factor NF-KB in brain microglia cells in culture (Olajide etal., 2022, Molecules. 27, 3342).

[0012] Q-actin™ is a cucumber extract standardised to 1% or greater content of the iminosugar acid idoBRI. Q-actin™ has been reported to be highly effective in clinical trials of sufferers of moderate to severe osteo-arthritis (Nash et al., 2023, Current Reviews in Rheumatology, 19, 345; Nash etal., 2018, Clin Interv Aging., 13, 2119) and performed better at 20mg daily than glucosamine and chondroitin at 2.7g per day. A more recent clinical study also confirmed the activity of cucumber extract at 20mg daily (Perez-Pinero et al. (2023) Appl. Sci, 13, 485) but the authors strangely concluded L-citrulline at 5% in a daily 20mg dose might be responsible for the biological effect even though the body produces L-citrulline itself and it is a common amino acid in foods; the authors did not appear to study idoBRI content.

[0013] The synthesis of idoBRI has been described in Fleet et al. (1986) Tetrahedron Lett. 27: 3205-3208. It can also be extracted from various botanical sources, including from parts of plants of the family Cucurbitaceae. It is a major component of certain cucumber fruits, and is the only iminosugar acid in cucumber. Thus, cucumbers from plants of the species Cucumis sativus are a good botanic source of idoBRI . Processes for enriching, purifying and extracting idoBRI from various botanical sources are described in WQ2021 / 019194 (the relevant teachings of which are incorporated herein by reference).

[0014] Melatonin and sleep

[0015] Sleep plays an important role in the maintenance of personal health. However, poor sleep quality and sleep deprivation are common nowadays. Various studies have demonstrated that inadequate sleep, such as short sleep duration or poor sleep quality, are associated with cardiovascular disease, obesity, hypertension, non-alcoholic liver disease, diabetes mellitus (DM), dyslipidemia, and even all-cause mortality (Chou etal., 2020, PLoS One, 15,9, :e0239185).

[0016] Melatonin (N-acetyl-5-methoxytryptamine) is well known to be important in regulating the sleep wake patterns (Braam etal., 2022, Sleep Medicine Reviews, 63, 101614). It is the main product of the pineal gland and after its formation, melatonin is released into the capillaries, and even in higher concentrations, into the cerebrospinal fluid and then spreads to most body tissues. The rhythm of melatonin production is primarily regulated by the suprachiasmatic nuclei (SCN) of the anterior hypothalamus. This ‘internal clock’ has a period which is not exactly 24 h. Information on the 24-h light and dark cycle is captured by retinal photo receptors and passed on to the SCN to reset this internal clock to a 24 h rhythm. There is a large inter-individual variation in the amount of melatonin synthesized in the glandula pinealis, as well as in the pineal volume. Although the amount of melatonin secretion shows great inter-individual variability, its range within the same person is highly reproducible from day to day, justifying singular melatonin measurements. More recently it has been found that melatonin is also synthesized in other organs (e.g., gastrointestinal tract), cells (e.g., leucocytes) and mitochondria (Tan et al., 2019, Melatonin research 2.1, 44). The amount of melatonin synthesized in the gastrointestinal tract can be more than a hundred times greater than in the pineal gland (Cipolla-Neto et al. 2018, Endocrine Reviews, 39, 990) but melatonin from extrapineal sites does not enter the circulation in substantial amounts.

[0017] The circadian rhythm of melatonin with high levels at night, and low levels during the day, reaches peak concentrations between 02:00 and 04:00 am. Longer nights are accompanied by a longer melatonin secretion. Melatonin production at night is not as efficient in the elderly population and clinical trials report that exogenous melatonin administration restores circadian disturbances.

[0018] To improve sleep quality there are medications available such as benzodiazepines, antidepressants, and antihistamines but due to adverse effects there is a trend towards alternative sleep aids such as natural and herbal therapies. Other food components such as glycine, theanine, and gamma-amino butyric acid (GABA) have been said to promote sleep but their potential applications are limited. For example, the effective timing of glycine intake is limited to before bedtime and its effective dose which is in the order of several grams; GABA has little brain penetration and the effective dose of theanine is several hundred milligrams, which is slightly lower than that of glycine (Katsube, 2022, J.

[0019] Functional Foods, 95, 105165 and papers cited therein). The same authors report that a mushroom sulfur-containing amino acid, ergothioneine, has strong antioxidant and antiinflammatory effects and can improve sleep in a clinical trial of people with high anxiety and sleep complaints. They also report that ergothioneine inhibits histamine N-methyltransferase and aldehyde dehydrogenase activity and binding to a3|34 nicotinic acetylcholine receptor and this might be related to the reported benefits to sleep. The common flavonoid luteolin is reported to have a hypnotic effect via Adenosine A1 and A2A Receptors (Kim, 2019, Biomol. Therapy, 27, 584); other flavonoids are also reported to have hypnotic effects but these are common in foods and so their contribution in the diet to sleep is not clear.

[0020] Sleep disorders have a profound negative impact on diverse somatic and psychological processes. Important symptoms and co-morbidities associated with sleep disorders include heart and vascular disease, hypertension, stroke, diabetes, hyperlipidemia, immunosuppression, obesity, metabolic syndrome, obstructive sleep apnea, inattention, decreased energy and fatigue. Effects on mental health include poor memory, depression, low mood and mood dysregulation, anxiety, stress, psychosis and problems with focus and concentration.

[0021] There remains an urgent need for alternative and improved treatments for sleep disorders.

[0022] Summary of the Invention

[0023] The present invention is based, at least in part, on the surprising discovery that idoBRI has melatonin stimulatory activity and can therefore be used in the treatment and prevention of sleep disorders.

[0024] The invention therefore finds application in the treatment and prevention of symptoms and co-morbidities associated with sleep disorders, including those selected from: heart and vascular diseases, hypertension, stroke, diabetes, hyperlipidemia, immunosuppression, obesity, metabolic syndrome, obstructive sleep apnea, inattention, decreased energy and fatigue, poor memory, depression, low mood and mood dysregulation, anxiety, stress, psychosis and problems with focus and / or concentration.

[0025] Thus, in a first aspect of the invention there is provided a composition comprising a compound of formula I:

[0026]

[0027] in which R represents H, or optionally substituted C1-15 alkyl, Ci.15 alkenyl or C1-15 alkynyl; or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, for use in the treatment or prevention of a sleep disorder.

[0028] In a related aspect, there is provided a compound of formula I:

[0029]

[0030] in which R represents H, or optionally substituted Ci.15 alkyl, Ci.15 alkenyl or C1-15 alkynyl; or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, for use in the treatment or prevention of a symptom or co-morbidity associated with a sleep disorder. In preferred aspects of this aspect of the invention, the symptom or co-morbidity associated with a sleep disorder may be selected from: heart disease, vascular disease, hypertension, stroke, diabetes, hyperlipidemia, immunosuppression, obesity, metabolic syndrome, obstructive sleep apnea, inattention, decreased energy, fatigue, poor memory, depression, low mood, mood dysregulation, anxiety, stress, psychosis and loss of focus and / or concentration.

[0031] In another related aspect, there is provided a compound of formula I:

[0032]

[0033] in which R represents H, or optionally substituted C1.15 alkyl, C1.15 alkenyl or C1.15 alkynyl; or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, for use in the treatment or prevention of a disease or disorder selected from: heart disease, vascular disease, hypertension, stroke, diabetes, hyperlipidemia, immunosuppression, obesity, metabolic syndrome, obstructive sleep apnea, inattention, decreased energy, fatigue, poor memory, depression, low mood, mood dysregulation, anxiety, stress, psychosis and loss of focus and / or concentration.

[0034] In some embodiments, the compound is of formula II:

[0035]

[0036] in which R represents H, or optionally substituted C1.15 alkyl, C1.15 alkenyl or C1.15 alkynyl; or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, for use in the treatment or prevention of a sleep disorder.

[0037] In preferred embodiments, the compound is idoBRI , or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof.

[0038] In particularly preferred embodiments, the compound is a pharmaceutically acceptable salt or solvate of idoBRI .

[0039] In another aspect, the invention relates to a method of treating or preventing a sleep disorder in a subject comprising the step of administering an effective amount of a compound as defined above, or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, to a subject in need thereof.

[0040] In preferred embodiments, the sleep disorder may be selected from insomnia, sleep apnea, restless leg syndrome, hypersomnia, narcolepsy, circadian rhythm disorders, parasomnia, non-REM sleep arousal disorders, REM sleep behaviour disorders, nightmare disorders, breathing-related sleep disorders, and substance- or medication-induced sleep disorder.

[0041] In certain embodiments, the sleep disorder is selected from insomnia related to: (a) depression; (b) anxiety; (c) PTSD; (d) schizophrenia; (e) Parkinson’s disease; (f) Alzheimer’s disease; (g) multiple sclerosis; (h) stroke; (i) pain; (j) shift work; (k) jet lag; (I) non-24 hour sleep-wake rhythm and / or (m) cancer.

[0042] Other aspects and preferred embodiments of the invention are defined and described in the claims set out below.

[0043] Detailed Description of the Invention

[0044] All publications, patents, patent applications and other references mentioned herein are hereby incorporated by reference in their entireties for all purposes as if each individual publication, patent or patent application were specifically and individually indicated to be incorporated by reference and the content thereof recited in full.

[0045] Definitions

[0046] Where used herein and unless specifically indicated otherwise, the following terms are intended to have the following meanings in addition to any broader (or narrower) meanings the terms might enjoy in the art:

[0047] Unless otherwise required by context, the use herein of the singular is to be read to include the plural and vice versa. The term "a" or "an" used in relation to an entity is to be read to refer to one or more of that entity. As such, the terms "a" (or "an"), "one or more," and "at least one" are used interchangeably herein. As used herein, the term "comprise," or variations thereof such as "comprises" or "comprising," are to be read to indicate the inclusion of any recited integer (e.g. a feature, element, characteristic, property, method / process step or limitation) or group of integers (e.g. features, element, characteristics, properties, method / process steps or limitations) but not the exclusion of any other integer or group of integers. Thus, as used herein the term "comprising" is inclusive or open-ended and does not exclude additional, unrecited integers or method / process steps.

[0048] The phrase "consisting essentially of’ is used herein to require the specified integer(s) or steps as well as those which do not materially affect the character or function of the claimed invention.

[0049] As used herein, the term "consisting" is used to indicate the presence of the recited integer (e.g. a feature, element, characteristic, property, method / process step or limitation) or group of integers (e.g. features, element, characteristics, properties, method / process steps or limitations) alone.

[0050] As used herein, the term disorder is used to define any abnormal condition that impairs physiological function and is associated with specific symptoms. The term is used broadly to encompass any disease, illness, abnormality, pathology, sickness, condition or syndrome in which physiological function is impaired irrespective of the nature of the aetiology (or indeed whether the aetiological basis for the disorder is established). It therefore encompasses conditions arising from infection, trauma, injury, surgery, radiological ablation, poisoning or nutritional deficiencies.

[0051] As used herein, the term treatment or treating refers to an intervention (e.g. the administration of an agent to a subject) which cures, ameliorates or lessens the symptoms of a disease or removes (or lessens the impact of) its cause(s) (for example, pathological variegated states). In this case, the term is used synonymously with the term “therapy”.

[0052] The term prevent or prevention refers to an intervention (e.g. the administration of an agent to a subject) which prevents or delays the onset or progression of a disorder or reduces (or eradicates) its incidence within a treated population. In this case, the term prevent or prevention is used synonymously with the term “prophylaxis”. The term subject (which is to be read to include "individual", "animal", "patient" or "mammal" where context permits) defines any subject, particularly a mammalian subject, for whom treatment is indicated. Mammalian subjects include, but are not limited to, humans, domestic animals, farm animals, zoo animals, sport animals and pet animals. In preferred embodiments, the subject is a human.

[0053] As used herein, an effective amount of a compound or composition defines an amount that can be administered to a subject without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio, but one that is sufficient to provide the desired effect, e.g. the treatment or prophylaxis manifested by a permanent or temporary improvement in the subject's condition. The amount will vary from subject to subject, depending on the age and general condition of the individual, mode of administration and other factors. Thus, while it is not possible to specify an exact effective amount, those skilled in the art will be able to determine an appropriate "effective" amount in any individual case using routine experimentation and background general knowledge. A therapeutic result in this context includes eradication or lessening of symptoms, reduced pain or discomfort, prolonged survival, improved mobility and other markers of clinical improvement. A therapeutic result need not be a complete cure.

[0054] The term bioisostere (or simply isostere) is a term of art used to define drug analogues in which one or more atoms (or groups of atoms) have been substituted with replacement atoms (or groups of atoms) having similar steric and / or electronic features to those atoms which they replace. The substitution of a hydrogen atom or a hydroxyl group with a fluorine atom is a commonly employed bioisosteric replacement. Sila-substitution (C / Si-exchange) is a relatively recent technique for producing isosteres. This approach involves the replacement of one or more specific carbon atoms in a compound with silicon (for a review, see Tacke and Zilch (1986) Endeavour, New Series 10: 191-197). The silasubstituted isosteres (silicon isosteres) may exhibit improved pharmacological properties, and may for example be better tolerated, have a longer half-life or exhibit increased potency (see for example Englebienne (2005) Med. Chem., 1(3): 215-226). Similarly, replacement of an atom by one of its isotopes, for example hydrogen by deuterium, may also lead to improved pharmacological properties, for example leading to longer half-life (see for example Kushner et al (1999) Can J Physiol Pharmacol. 77(2):79-88). In its broadest aspect, the present invention contemplates all bioisosteres (and specifically, all silicon bioisosteres) of the compounds of the invention. The term pharmaceutically acceptable salt defines any non-toxic organic or inorganic acid addition salt of the free base which are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and which are commensurate with a reasonable benefit / risk ratio. Suitable pharmaceutically acceptable salts are well known in the art. Examples are the salts with inorganic acids (for example hydrochloric, hydrobromic, sulphuric and phosphoric acids), organic carboxylic acids (for example acetic, propionic, glycolic, lactic, pyruvic, malonic, succinic, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, dihydroxymaleic, benzoic, phenylacetic, 4-aminobenzoic, 4-hydroxybenzoic, anthranilic, cinnamic, salicylic, 2-phenoxybenzoic, 2-acetoxybenzoic and mandelic acid) and organic sulfonic acids (for example methanesulfonic acid and p-toluenesulfonic acid).

[0055] These salts and the free base compounds can exist in either a hydrated or a substantially anhydrous form. Crystalline forms, including all polymorphic forms, of the iminosugars of the invention are also contemplated and in general the acid addition salts of the compounds are crystalline materials which are soluble in water and various hydrophilic organic solvents and which in comparison to their free base forms, demonstrate higher melting points and an increased solubility.

[0056] The term pharmaceutically acceptable metabolite as applied to the compounds of the invention defines a pharmacologically active product produced through metabolism in the body of the specified compound or salt thereof.

[0057] The term pharmaceutically acceptable prodrug as applied to the compounds of the invention defines any pharmaceutically acceptable compound that may be converted under physiological conditions or by solvolysis to the specified compound, to a pharmaceutically acceptable salt of such compound or to a compound that shares at least some of the activity of the specified compound. Particularly preferred pro-drugs are ester derivatives which are esterified at one or more of the free hydroxyls and which are activated by hydrolysis in vivo. Derivatization may also augment other biological activities of the compound, for example bioavailability and / or glycosidase inhibitory profile. For example, derivatization may increase CNS penetration (e.g. penetration of the blood-brain barrier). Prodrugs and active metabolites of the compounds of the invention may be identified using routine techniques known in the art (see for example, Bertolini etal., J. Med. Chem., 1997, 40, 2011-2016).

[0058] In the present specification the term “alkyl” defines a straight or branched saturated hydrocarbon chain. The term “Ci-C6alkyl” refers to a straight or branched saturated hydrocarbon chain having one to six carbon atoms. The term “C1-C9 alkyl” refers to a straight or branched saturated hydrocarbon chain having one to nine carbon atoms. The term “C1-C15 alkyl” refers to a straight or branched saturated hydrocarbon chain having one to fifteen carbon atoms. Preferred is Ci-C6alkyl. Examples include methyl, ethyl, n-propyl, isopropyl, t-butyl, n-hexyl. The alkyl groups of the invention may be optionally substituted by one or more halogen atoms.

[0059] In the present specification the term “alkenyl” defines a straight or branched hydrocarbon chain having containing at least one carbon-carbon double bond. The term “Ci-C6alkenyl” refers to a straight or branched unsaturated hydrocarbon chain having one to six carbon atoms. The term “C1-C9 alkenyl” refers to a straight or branched unsaturated hydrocarbon chain having one to nine carbon atoms. The term “C1-C15 alkenyl” refers to a straight or branched unsaturated hydrocarbon chain having one to fifteen carbon atoms. Preferred is Ci-C6alkenyl. Examples include ethenyl, 2-propenyl, and 3-hexenyl. The alkenyl groups of the invention may be optionally substituted by one or more halogen atoms.

[0060] In the present specification the term “alkynyl” defines a straight or branched hydrocarbon chain having containing at least one carbon-carbon triple bond. The term “Ci-C6alkynyl” refers to a straight or branched unsaturated hydrocarbon chain having one to six carbon atoms. The term “C1-C9 alkynyl” refers to a straight or branched unsaturated hydrocarbon chain having one to nine carbon atoms. The term “C1-C15 alkynyl” refers to a straight or branched unsaturated hydrocarbon chain having one to fifteen carbon atoms. Preferred is Ci-C6alkynyl. Examples include ethynyl, 2-propynyl, and 3-hexynyl. The alkynyl groups of the invention may be optionally substituted by one or more halogen atoms.

[0061] The term isolated as applied to the idoBRI of the invention is used herein to indicate that the idoBRI exists in a physical milieu distinct from that in which it occurs in nature (or in the case of synthetic idoBRI, is purified to some degree). For example, the isolated idoBRI may be substantially isolated (for example purified) with respect to the complex cellular milieu in which it naturally occurs (or with respect to the some or all of the starting products, intermediates, buffers, solvents, reactants and / or co-products from which it is synthesised).

[0062] When the isolated material (e.g. synthetic, non-naturally occurring idoBRI) is purified, the absolute level of purity is not critical and those skilled in the art can readily determine appropriate levels of purity according to the use to which the material is to be put.

[0063] Preferred, however, are purity levels of 50% w / w, 60% w / w, 70% w / w, 80% w / w, 90% w / w, 99% w / w or higher. In some circumstances, the isolated idoBRI forms part of a composition (for example a more or less crude extract containing many other substances) or buffer system, which may for example contain other components. In other circumstances, the isolated idoBRI may be purified to essential homogeneity, for example as determined spectrophotometrically, by NMR or by chromatography (for example GC-MS of the trimethylsilyl-derivatives) .

[0064] The compounds of the present invention can be administered by oral or parenteral routes. Preferred is oral administration. The amount administered can vary widely according to the particular dosage unit employed, the period of treatment, the age and sex of the patient treated, the nature and severity of the sleeping disorder treated, and the particular compound selected.

[0065] Moreover, the compounds of the invention can be used in conjunction with other agents known to be useful in the treatment or prevention of sleeping disorders, and in such embodiments the dose may be adjusted accordingly.

[0066] In general, the effective amount of the compound administered will generally range from about 1 pg / kg to 500 mg / kg daily. A unit dosage may contain from 0.05 to 500 mg of the compound, and can be taken one or more times per day. The compound can be administered with a pharmaceutical carrier using conventional dosage unit forms, as described below.

[0067] The preferred route of administration is oral administration. In general, a suitable dose for human subjects will be in the range of 0.1 pg / kg to 10 pg / kg body weight of the subject per day. Thus, in general, the idoBRI, or pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, is orally administered at a dose of at least 0.1 mg prior to bedtime, for example at a dose of 0.1 to 10 mg prior to bedtime, for example 0.1 to 5 mg prior to bedtime, or about 0.2 mg prior to bedtime.

[0068] The desired dose is preferably presented as a single dose for daily administration.

[0069] However, two, three, four, five or six or more sub-doses administered at appropriate intervals throughout the day may also be employed. These sub-doses may be administered in unit dosage forms.

[0070] Formulation

[0071] The compound for use according to the invention may take any form. It may be synthetic or isolated from natural sources (for example from any of the botanical sources identified herein, including for example a botanical source selected from plants of the family Cucurbitaceae (for example, plants of the genus Cucumis, e.g. plants of the species Cucumis sativus). Particularly preferred as botanical source are fruits of the foregoing plants, for example cucumbers, pumpkins, squashes or gourds. The term isolated as applied to the compounds of the invention is used herein to indicate that the compound exists in a physical milieu distinct from that in which it occurs in nature. For example, the isolated compound may be substantially isolated (for example enriched or purified) with respect to the complex cellular milieu in which it naturally occurs. The isolated compound may therefore take the form of an enriched fraction or extract of any of the botanical sources described herein. When the isolated material is enriched or purified, the absolute level of enrichment or purity is not critical and those skilled in the art can readily determine appropriate levels according to the use to which the material is to be put. Preferred are purity levels of at least 0.1% w / w, 0.2% w / w, 0.3% w / w, 0.4% w / w, 0.5% w / w, 0.6% w / w, 0.7% w / w, 0.8% w / w, 0.9% w / w, 1.0% w / w, 1.1% w / w, 1.2% w / w, 1.3% w / w, 1.4% w / w, 1.5% w / w, 1.6% w / w, 1.7% w / w, 1.8% w / w, 1.9% w / w or 2.0% w / w. Particularly preferred are purity levels of at least 0.5-2.0% w / w, for example at least 0.8-1.5% w / w, for example at least about 1.0% w / w. Levels of 5-10% w / w may be readily obtained in cases where the material is isolated from natural sources, if necessary, by employing suitable enrichment techniques, such as ion exchange chromatography.

[0072] In some circumstances, the isolated compound forms part of a composition (for example a more or less crude extract containing many other substances) or buffer system, which may for example contain other components. In other circumstances, the isolated compound may be purified to essential homogeneity, for example as determined spectrophotometrically, by NMR or by chromatography (for example GC-MS of the trimethylsilyl-derivatives).

[0073] When isolated from a natural source, the idoBRI may be purified. However, the compositions of the invention may take the form of herbal medicines. The term herbal medicine is used herein to define a pharmaceutical composition in which at least one active principle (e.g. the compound) is not chemically synthesized and is a phytochemical constituent of a plant. In most cases, this non-synthetic active principle is not isolated (as defined herein), but present together with other phytochemicals with which it is associated in the source plant. In some cases, however, the plant-derived bioactive principle(s) (for example, idoBRI) may be in a concentrated fraction or isolated (sometimes involving high degrees of purification). In many cases, however, the herbal medicine comprises a more or less crude extract, infusion or fraction of a plant or even an unprocessed whole plant (or part thereof), though in such cases the plant (or plant part) is usually at least dried and / or milled. The herbal medicine may be in the form of a food supplement, food additive, nutraceutical, a beverage or presented in unitary doses as a herbal pharmaceutical kit or pack. Such herbal medicines preferably are analysed to determine whether they meet a standard specification prior to use. The term standard specification defines a characteristic, or a phytochemical profile, which is correlated with an acceptable quality of the herbal medicine, cosmetic or nutraceutical. In this context, the term quality is used to define the overall fitness of the product for its intended use, and includes the activity of idoBRI at an appropriate concentration.

[0074] Herbal medicines suitable for use according to the invention may be dried plant material. Alternatively, the herbal medicine may be processed plant material, the processing involving physical or chemical pre-processing, for example powdering, grinding, freezing, evaporation, filtration, pressing, spray drying, extrusion, supercritical solvent extraction and tincture production. In cases where the herbal medicine is administered or sold in the form of a whole plant (or part thereof), the plant material may be dried prior to use. Any convenient form of drying may be used, including freeze-drying, spray drying or air-drying.

[0075] IdoBRI may be separated from the higher molecular weight components such as proteins and polysaccharides by using various membrane technologies. These include microfiltration, ultrafiltration and nanofiltration. Alternatively, or in addition, electrodialysis may also be used to concentrate the charged idoBRI . These methods use membranes of pore sizes that allow only molecules below a certain size to pass or rely on charges on the molecules to allow or not allow them to pass through the membrane. Anion and cation exchange resins may also be used to concentrate the idoBRI .

[0076] When isolated from a natural source, the compound for use according to the invention may be purified. In embodiments where the compound is formulated together with a pharmaceutically acceptable excipient, any suitable excipient may be used, including for example inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavouring agents, colouring agents and preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while corn starch and alginic acid are suitable disintegrating agents. Binding agents may include starch and gelatin, while the lubricating agent, if present, will generally be magnesium stearate, stearic acid or talc.

[0077] The pharmaceutical compositions may take any suitable form, and include for example gummies, tablets, elixirs, capsules, solutions, suspensions, powders, granules and aerosols.

[0078] Tablets for oral use may include the compound for use according to the invention, mixed with pharmaceutically acceptable excipients, such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavouring agents, colouring agents and preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while corn starch and alginic acid are suitable disintegrating agents. Binding agents may include starch and gelatin, while the lubricating agent, if present, will generally be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate, to delay absorption in the gastrointestinal tract. Capsules for oral use include hard gelatin capsules in which the compound for use according to the invention is mixed with a solid diluent, and soft gelatin capsules wherein the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin or olive oil. For oral administration the compound can be formulated into solid or liquid preparations such as gummies, capsules, pills, tablets, troches, lozenges, melts, powders, granules, solutions, suspensions, dispersions or emulsions (which solutions, suspensions dispersions or emulsions may be aqueous or non-aqueous). The solid unit dosage forms can be a capsule which can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers such as lactose, sucrose, calcium phosphate, and cornstarch.

[0079] In another embodiment, the compounds of the invention are tableted with conventional tablet bases such as lactose, sucrose, and cornstarch in combination with binders such as acacia, cornstarch, or gelatin, disintegrating agents intended to assist the break-up and dissolution of the tablet following administration such as potato starch, alginic acid, corn starch, and guar gum, lubricants intended to improve the flow of tablet granulations and to prevent the adhesion of tablet material to the surfaces of the tablet dies and punches, for example, talc, stearic acid, or magnesium, calcium, or zinc stearate, dyes, colouring agents, and flavouring agents intended to enhance the aesthetic qualities of the tablets and make them more acceptable to the patient.

[0080] Suitable excipients for use in oral liquid dosage forms include diluents such as water and alcohols, for example, ethanol, benzyl alcohol, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptably surfactant, suspending agent or emulsifying agent.

[0081] Exemplification

[0082] The invention will now be described with reference to specific Examples. These are merely exemplary and for illustrative purposes only: they are not intended to be limiting in any way to the scope of the monopoly claimed or to the invention described. These examples constitute the best mode currently contemplated for practicing the invention.

[0083] Example: Randomized, placebo controlled clinical human trial

[0084] Introduction

[0085] A randomized, placebo controlled clinical human trial of an idoBRI dosage form (2 x gummies daily, each gummy containing 10mg of a standardised extract >1% idoBRI) or placebo (2 x gummies with no idoBRI , taken twice daily before bed), in middle aged and older adults (>50), was carried out for a period of 12 weeks. The aim was to explore whether consumption of idoBRI had any impact on sleep and health related parameters including cognition and manual dexterity. Home-urine samples were collected for the investigation into the chemical composition. The trial took place over two sites in Wales in 2023.

[0086] Design

[0087] IdoBRI composition tested

[0088] The idoBRI dosage form (2 x gummies, each gummy containing 10mg of a standardised extract >1% idoBRI) was consumed daily before bed. The placebo was a gummy matching the idoBRI dosage form in appearance, taste and texture. The gummies were wax coated, square bottom and dark red in colour with natural raspberry flavouring. Vegan gummies were chosen as opposed to tablets because they are easier to consume than pills / capsules. Both the idoBRI dosage form and the placebo were manufactured by a fully licensed GMP site manufacturer. The gummies were analysed and quality control screened for elemental elements and microbials and passed all according to guidelines.

[0089] Product safety

[0090] The Food Standards Agency UK confirmed that the idoBRI dosage form (Q-actin™) can be used in clinical trials. A GRAS document (Generally Recognised As Safe) status has been prepared for the producers in America (Gateway Health Alliances, Inc. / lminoTech, Inc.) by Soni & Associates Inc. The gummy ingredients were: Corn Syrup, Sugar, Purified Water, Pectin, Natural Flavours (raspberry), Citric Acid, MCT Oil, Natural Colours (vegetable concentrates), Carnauba Wax, Sodium Citrate.

[0091] Outline of Trial

[0092] This was a randomised, double-blinded, parallel study, where participants were asked to take the gummies (the idoBRI dosage form or placebo) twice a day for a period of 12 weeks. Within that 12 week period, participants were required to make three visits to the Well-being and Health Assessment Research Unit (WARU) at Aberystwyth University, or to the remote facility in Trimsaran, following a pre-induction screening visit. Urine kits and study materials were prepared (and stored) at the Well-Being and Health Assessment Research Unit (WARU) at Aberystwyth University. Urine samples were assessed at Aberlnnovation and Aberystwyth University using high resolution metabolomics. Self-report data was analysed in WARU using appropriate methods.

[0093] 49 participants started the trial; 33 were female, and 16 were male. The age range was between 50 and 78 years old, with a mean age of 63.02. Most participants were healthy in general and did not highlight any health problems when recording their medical history. Body mass index (BMI) varied between age and gender, however, mean BMI (50.35 kg nr2) based on all 49 participants who provided both weight and height measurements, indicated that the cohort showed all participants scoring in the obese and severely obese categories (World Health Organization, 2006). Waist to hip ratio (WHR) was calculated to highlight those with increased risk of developing a metabolic complication. On average females had a WHR of 0.90, which according to guidelines indicates high health risk (>0.86). Males had an average WHR of 1.02 which according to guidelines indicates high health risk.

[0094] There was no financial incentive for participants to join the study. This was the first time this type of research was conducted in a healthy ageing cohort. Ethical approval was obtained from the Aberystwyth University Research Ethics Panel.

[0095] • Recruited: 47

[0096] • Aberystwyth: 15 | Trimsaran: 32

[0097] • Drop-outs: Nine

[0098] Week O

[0099]

[0100] • Mean Age: 62.75

[0101] Week 12

[0102] • Males: 12 | Females: 26

[0103] • Mean Age: 61.50

[0104] Week O

[0105] Condition A (Intervention, idoBRI): 25 Condition B (Placebo): 22

[0106] Week 12

[0107] • Condition A (Intervention, idoBRI): 19

[0108] • Condition B (Placebo): 19

[0109] Urine

[0110] Chemical composition analysis used Flow Infusion Electrospray Ionisation Mass Spectrometry (FIE-MS). The FIE-MS analysis was conducted with an Exploris 120 mass analyser coupled with a Dionex Vanquish UHPLC system (Thermo-Scientific) using RP-C18, measuring ion intensities within the m / z range of 55 to 1200. The top-ranked features contributing to multi-dimensional scaling models were identified using re-sampling methods and p-values for False Positive Rates (FPR < 0.05). Features that showed good classification performance in pairwise comparisons were further investigated. All p-values were corrected for multiple testing using the Bonferroni correction. Tentatively identified melatonin derivatives were found to be elevated after idoBRI dosage form supplementation, suggesting a positive effect on sleep quality, which correlated with selfreported Pittsburgh Sleep Quality Index. The relative m / z-transition areas of melatonin and its two main biotransformation products in selected urine samples after the idoBRI dosage form supplementation were examined following LC-MS / MS fragmentation on the Exploris-120, revealing interesting trends. Authentic reference compounds were purchased to confirm the identifications.

[0111] Results

[0112] Most notable were changes to sleep quality (Pittsburgh Sleep Quality Index (PSQI)). Surprisingly, sleep related parameters showed a trend to improvement that was correlated with an observation that levels of melatonin and related derivatives such as 6-Sulfatoxymelatonin and 6-Hydroxymelatonin were greatly increased in urine of those taking idoBRI relative to those in the placebo group. Melatonin is hydroxylated to 6-hydroxymelatonin, followed by sulfate conjugation to 6-hydroxymelatonin sulfate (90%) or glucuronide conjugation to 6-hydroxymelatonin glucuronide (10%), with approximately 5% of serum melatonin being excreted unmetabolized through urine. Lower nocturnal melatonin levels are associated with worsened daytime sleepiness, sleep efficiency, and shorter sleep time in older men. Saksvik-Lehouillier, I., et al., 2015, The Journals of Gerontology, A, 70, (12), 1569).

[0113] After correcting for Greenhouse-Geisser following a violation of sphericity, there was a nonsignificant finding for total PSQI score both within-and-between groups. This suggests that consuming idoBRI for 12 weeks did not significantly improve sleep. However, the decreasing trend for group A suggests that a longer intervention period or higher dose might significantly improve sleep quality overall.

[0114] Table showing mean PSQI sleep quality score for the two conditions at 0, 6 and 12 weeks

[0115]

[0116] Errors ± 2 SE

[0117] After correcting for Greenhouse-Geisser following a violation of sphericity, there was a nonsignificant main effect on sleep latency, and a non-significant interaction between the two groups. However, the decreasing trend in PSQI_C2 scores for the experimental group suggests that a longer intervention period could significantly improve sleep latency.

[0118] Table showing mean PSQI_2 sleep latency score for the two conditions at 0, 6 and 12 weeks

[0119]

[0120] Errors ± 2 SE

[0121] Looking just at the sub-group over 60 years of age, there was also a non-significant finding for sleep efficiency between weeks 0, 6, and 12, within both groups. This suggests that consuming idoBRI for 12 weeks did not significantly improve sleep efficiency. However, the trending improvement in sleep efficiency (whilst not significant), suggests that a trial duration exceeding 12 weeks could yield significant improvements in sleep efficiency in the future, in people over 60.

[0122] Table showing mean PSQI_C4 sleep efficiency score in over 60s for the two conditions at 0, 6 and 12 weeks

[0123]

[0124] Errors ± 2 SE

[0125] Again looking just at the over 60 years of age participants, there was a non-significant finding for sleep disturbance both within-and-between groups but consuming idoBRI for 12 weeks did give a trend to improved sleep disturbance scores in people over 60 but not statistically significant.

[0126] Table showing mean PSQI_C5 sleep disturbance score in the over 60s for the two conditions at 0, 6 and 12 weeks

[0127]

[0128] Errors ± 2 SE

[0129] Mass spectral urine analysis

[0130] From the FIE-MS relative response data it was seen there was an increase in melatonin and metabolites in participants taking idoBRI but the largest increase was in 6-sulfatoxy-melatonin. The placebo group did not show the increase in melatonin and metabolites. A subset of the low, medium and high melatonin samples were selected for quantification using a TSQ Quantis using RP-C18 and authentic standards. There were extremely low levels of 6-hydroxymelatonin detected in the 'medium' and 'high' samples, with slightly higher concentrations of melatonin overall. Neither of these were detectable in the 'low' samples (consumed no idoBRI-containing gummies). The proportions of melatonin and 6-hydroxymelatonin appear to rise in step with 6-sulfatoxy-melatonin in the arbitrary ranking of low, medium and high. The same urine extracts were run on both instruments, the TSQ Quantis data agrees with the Exploris results although the Exploris is more sensitive.

[0131] Table showing analysis data from samples selected from preliminary FIE-MS data for 6-sulfatoxy-melatonin - 2 x high, 2 x medium and 2 x low values.

[0132] All samples were adjusted for Rl using water, then extracted 50:50 with MeOH.

[0133]

[0134] Table of the results for two participants showing the increasing relative levels of 6-sulfatoxy-melatonin from FIE-MS data.

[0135]

[0136]

[0137] Improved sleep correlated with improved cognition and manual dexterity

[0138] Sleep disorders negatively impact both physical and mental performance. The ability of idoBRI to ameliorate these clinical sequelae was studied by measuring finger dexterity in the Nine-Hole Peg Test (9HPT).

[0139] In the idoBRI group, participants showed a significant improvement between week 6 and week 12, achieving a faster peg test score on average. In comparison, participants in the placebo group showed no significant changes between weeks 0, 6, and 12, suggesting that consumption of idoBRI for 12 weeks significantly improved dexterity.

[0140] Table showing mean Peg Test speed score for the two conditions at 0, 6 and 12 weeks

[0141]

[0142] Errors ± 2 SE

[0143] Statistical analysis of the Peg Test data

[0144] > >

[0145] >

[0146] >

[0147]

[0148]

[0149] Conclusion

[0150] Administration of an idoBRI composition gave a trend over 12 weeks to improved sleep in healthy people aged over 50 compared to a group receiving a placebo. The trend to improved sleep was correlated with a greatly increased level of melatonin and metabolites measured in urine in the first morning void, and also with improved cognition and manual dexterity.

[0151] Equivalents

[0152] The foregoing description details presently preferred embodiments of the present invention. Numerous modifications and variations in practice thereof are expected to occur to those skilled in the art upon consideration of these descriptions. Those modifications and variations are intended to be encompassed within the claims appended hereto.

Claims

CLAIMS1. A composition comprising a compound of formula I:in which R represents H, or optionally substituted C1.15 alkyl, C1.15 alkenyl or C1.15 alkynyl; or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, for use in the treatment or prevention of a sleep disorder.

2. A composition as defined in claim 1 for use in the treatment or prevention of a symptom or co-morbidity associated with a sleep disorder, optionally selected from: heart disease, vascular disease, hypertension, stroke, diabetes, hyperlipidemia, immunosuppression, obesity, metabolic syndrome, obstructive sleep apnea, inattention, decreased energy, fatigue, poor memory, depression, low mood, mood dysregulation, anxiety, stress, psychosis and loss of focus and / or concentration.

3. The composition for use according to claim 1 or claim 2, wherein the compound is of formula II:in which R represents H, or optionally substituted C1.15 alkyl, C1.15 alkenyl or C1.15 alkynyl; or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, for use in the treatment or prevention of a sleep disorder.

4. The composition for use according to any one of the preceding claims, wherein the compound is idoBRI, or a pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof.

5. The composition for use according to any one of the preceding claims, wherein the compound is a pharmaceutically acceptable salt or solvate of idoBRI .

6. The composition for use according to any one of the preceding claims, wherein the sleep disorder is selected from insomnia, delayed sleep phase syndrome, sleep apnea, restless leg syndrome, hypersomnia, narcolepsy, circadian rhythm disorders (including non-24-hour sleep wake disorder), parasomnia, non-REM sleep arousal disorders, REM sleep behaviour disorders, nightmare disorders, breathing-related sleep disorders, and substance- or medication-induced sleep disorder.

7. The composition for use according to claim 6, wherein the sleep disorder is selected from insomnia related to: (a) depression; (b) anxiety; (c) PTSD; (d) schizophrenia; (e) Parkinson’s disease; (f) Alzheimer’s disease; (g) multiple sclerosis; (h) stroke; (i) pain; (j) shift work; (k) jet lag; (I) non-24 hour sleep-wake rhythm and / or (m) cancer.

8. The composition for use according to any one of the preceding claims, wherein the compound, or pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, is present in the composition at a level of at least: 0.1% w / w, 0.5% w / w, 1% w / w, 5% w / w, 10% w / w; 15% w / w; 20% w / w; 25% w / w; 30% w / w; 35% w / w; 40% w / w; 45% w / w; 50% w / w, 60% w / w, 70% w / w, 80% w / w, 90% w / w, 99% w / w (on a dry weight basis).

9. The composition for use according to any one of the preceding claims, wherein the compound, or pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, is present in the composition at a level of at least 1% w / w (on a dry weight basis).

10. The composition for use according to any one of the preceding claims, wherein the composition is administered orally.

11. The composition for use according to claim 10, wherein the compound is idoBRI , or a pharmaceutically acceptable salt or solvate thereof, and is administered at a dose of at least 0.1 mg prior to bedtime.

12. The composition for use according to claim 10 or 11, wherein the compound is idoBRI , or a pharmaceutically acceptable salt or solvate thereof, and is administered at a dose of 0.1 to 10 mg prior to bedtime, for example 0.1 to 5 mg prior to bedtime, or about 0.2 mg prior to bedtime.

13. A method of: (a) treating or preventing a sleep disorder in a subject; or (b) of treating or preventing a symptom or co-morbidity associated with a sleep disorder, optionally selected from: heart disease, vascular disease, hypertension, stroke, diabetes, hyperlipidemia, immunosuppression, obesity, metabolic syndrome, obstructive sleep apnea, inattention, decreased energy, fatigue, poor memory, depression, low mood, mood dysregulation, anxiety, stress, psychosis and loss of focus and / or concentration, comprising the step of administering an effective amount of a compound as defined in any one of claims 1-5, ora pharmaceutically acceptable salt, solvate, metabolite, prodrug, bioisostere, or protected form thereof, to a subject in need thereof.

14. The method of claim 13, wherein the sleep disorder is as defined in claim 6 or 7.

15. The method of claim 13 or 14, wherein: (a) the compound is present in the composition at a level as defined in claim 8 or 9; and / or (b) the composition is administered as defined in any one of claims 10-12.

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