Naringin or naringenin for improving sleep quality

Abstract

The present invention relates to flavonoids for use in improving sleep quality. The flavonoids of the invention are naringin or naringenin and hesperidin. The compounds are particularly useful in decreasing sleep onset latency, decreasing sleep latency, improving sleep efficiency, decreasing sleep disturbance, decreasing wake after sleep onset, decreasing the number of awakenings, improving the ratio of sleep stages, decreasing medication use, decreasing daytime dysfunction, feeling rested / refreshed, and feeling sleepy after waking up.

Description

[0001] NARINGIN OR NARINGENIN FOR IMPROVING SLEEP QUALITY

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to flavonoids for use in improving sleep quality. The flavonoids of the invention are in particular naringin or naringenin and hesperidin. The compounds are particularly useful in decreasing sleep onset latency, decreasing sleep latency, improving sleep efficiency, decreasing sleep disturbance, decreasing wake after sleep onset, decreasing the number of awakenings, improving the ratio of sleep stages, decreasing medication use, decreasing daytime dysfunction, feeling rested / refreshed, and feeling sleepy after waking up.

[0004] BACKGROUND OF THE INVENTION

[0005] Sleep is a vital aspect of overall well-being, influencing physical, mental, and emotional health. However, an increasing number of individuals are grappling with sleep problems that negatively impact their daily lives. Addressing these issues and improving sleep quality is essential for maintaining optimal health and productivity. Sleep disorders are conditions that affect the quality, amount and timing of sleep. Common sleep disorders include insomnia, restless legs syndrome, narcolepsy and sleep apnea. Sleep disorders can affect mental and physical health.

[0006] Self-help strategies include structuring sleep schedules, calming bedtime routine, and optimizing the sleep environment by for instance morning light exposure and evening dim light. However, self-help strategies do not always alleviate sleep disorders and individual responses to treatments vary.

[0007] Medications can help improving sleep quality, especially for certain sleep disorders or short-term sleep disturbances. However, medication should be considered under the guidance and supervision of a healthcare professional, particularly a doctor specializing in sleep medicine, since patients may become dependent or addicted. These medications help induce and maintain sleep. They include benzodiazepines (e.g., temazepam, lorazepam) and non-benzodiazepines (e.g., zolpidem, eszopiclone). Another example is melatonin supplementation, which can help regulate sleep-wake cycles. Further, dopamine agonist can help relieve the symptoms of restless leg syndrome by increasing dopamine levels in the brain. Also, stimulant medication such as modafinil and armodafinil are often prescribed to improve wakefulness and reduce excessive daytime sleepiness. Sodium oxybate can improve nighttime sleep and reduce daytime sleepiness in narcolepsy patients.

[0008] Each of these medications may have side effects or interactions, and their long-term use requires careful monitoring. The search for sleep medication which does not cause side effects is thus on- going. The goal of the present invention is therefore to provide alternatives to classical sleep medication for increasing sleep quality while inducing minimal side effects.

[0009] The inventors surprisingly found that Hesperidin and naringin or hesperidin and naringenin has a beneficial effect on sleep quality. The present invention therefore relates to the use of hesperidin and naringin or hesperidin and naringenin for improving sleep quality, wherein naringin or naringenin is administered via oral administration. More specifically a composition comprising hesperidin and naringin or hesperidin and naringenin and hesperidin has a beneficial effect on sleep quality, wherein the composition is administered via oral administration.

[0010] In addition, oral administration of hesperidin and naringin or hesperidin and naringenin can be performed without medical supervision. It is available as supplement and no side effects are observed when hesperidin and naringin or hesperidin and naringenin is administered on daily basis. Hesperidin and naringin or hesperidin and naringenin for use in improving sleep quality, when administered orally, is therefore advantageous over traditional medication.

[0011] SUMMARY OF THE INVENTION

[0012] It has surprisingly been found that compounds such as hesperidin and naringin or hesperidin and naringenin and close analogues can be used to improve sleep quality in different subjects, wherein hesperidin and naringin or hesperidin and naringenin is administered via oral administration. In addition, improvements are observed when a flavanone of the naringin or naringenin-like group is combined with hesperidin. Although not required, hesperidin and naringin or hesperidin and naringenin can be obtained easily from natural products, such as citrus fruits.

[0013] Naringin is a flavanone, a subclass of the flavonoids found in grapes and citrus fruits. Naringin is known to possess numerous biological properties such as antioxidant, anti-inflammatory, anti- apoptotic and anti-carcinogenic activities (e.g. WO200126467, US2002037855, WO05094864A2, etc.). Naringin is metabolized to its aglycone naringenin by the gut microbiota, resulting in an increased bioavailability.

[0014] Hesperidin (also referred to herein as hesperetin-7-rutinoside) is a flavanone, a subclass of the flavonoids that is most abundant in the fruit of immature citrus fruits and in the peels of mature citrus fruits, and exhibits a wide range of physiological activity. Hesperidin is known to have vasodilative, hypotensive, anti-oxidative, anti-inflammatory, carcinostatic and prebiotic activities (e.g. EP0956867, WO199847534, WO200115553, US2002037855, US2003078231, etc.). Hesperidin is metabolized to its aglycone hesperetin by the gut microbiota, resulting in an increased bioavailability.

[0015] Some particular health benefits attributed to citrus consumption have been correlated with the presence of some specific flavonoids. Flavonoids are polyphenolic compounds with diverse beneficial biochemical effects, including anti-oxidative, and anti-inflammatory activities that can modulate immune responses.

[0016] Here, the compounds described herein for use in the invention may originate from any plant and may refer to any of the following: 1) the native glycoside naringin as well as partially deglycosylated forms or its metabolized aglycone naringenin, and further metabolized derivatives that can result from bacterial digestion, intestinal uptake, and hepatic metabolism. They may further comprise any chemically or enzymatically derived derivative of any of the foregoing molecules. 2)_the native glycoside hesperidin as well as partially deglycosylated forms, the aglycon form hesperetin, and further metabolized derivatives that can result from bacterial digestion, intestinal uptake, and hepatic metabolism. They may further comprise any chemically or enzymatically derived derivative of any of the foregoing molecules.

[0017] The inventors observed a synergistic effect when naringin or naringenin is combined with hesperidin. Therefore, the present invention provides the use of a composition comprising hesperidin and naringin or hesperidin and naringenin and for improving sleep quality, wherein the composition is administered via oral administration. In addition, the present invention relates to hesperidin and naringin or hesperidin and naringenin or a composition as described herein comprising these combinations of compounds for use in improving sleep quality, wherein the composition is administered via oral administration.

[0018] In another particular embodiment, the composition as described herein further comprises isonaringin, neoponcirin and / or hesperetin.

[0019] In a further embodiment, the invention relates to the use of a composition comprising naringin and hesperidin or naringenin and hesperidin, and optionally isonaringin, neoponcirin and / or hesperetin for improving sleep quality, wherein sleep quality is defined according to any of the group consisting of: decreased sleep onset latency, decreased sleep latency, improved sleep efficiency, decreased sleep disturbance, decreased wake after sleep onset, decreased number of awakenings, improved ratio of sleep stages, decreased medication use, decreased daytime dysfunction, feeling rested / refreshed, and feeling less sleepy after waking up. In another embodiment, the invention relates to the use of a composition as described herein for improving sleep quality, wherein naringin or naringenin is present in at least 0.1 wt.%, preferably in at least 1 wt.%, more preferably in at least 2 wt.%, even more preferably in between 3 and 7 wt.%, most preferably in about 5 wt.%.

[0020] Additionally, the invention relates to the use of a composition as described herein for improving sleep quality, wherein hesperidin is present in at least 30 wt.%, preferably in at least 50 wt.%, more preferably in at least 70 wt.%, even more preferably in between 80 and 90 wt.%, most preferably in about 84 wt.%.

[0021] Furthermore, the invention relates to the use of a composition as described herein, wherein the composition is a pharmaceutical composition, optionally comprising an excipient or pharmaceutically acceptable carrier.

[0022] Additionally, the invention relates to the use of a composition as described herein, wherein the use is a non-therapeutic use.

[0023] In a further aspect the invention relates to the use of a composition as described herein, wherein said composition is in the form of a liquid, solution, tablet, pill, powder, lozenge, dissolvable film, suspension, dietary gel, capsule, chewable, gummy or syrup.

[0024] In a specific embodiment, the invention relates to the use of a composition as described herein for improving sleep quality, wherein hesperidin is enantiomerically enriched for 2S-hesperidin and in a more specific embodiment, the invention relates to the use of a composition as described herein for improving sleep quality, wherein at least 60%, more preferably 70%, of hesperidin is 2S- Hesperidin.

[0025] Another aspect of the present invention is a method for improving sleep quality in a subject comprising a step of administering hesperidin and naringin or hesperidin and naringenin or a composition as described herein to a subject, wherein the hesperidin and naringin or hesperidin and naringenin or the composition is administered via oral administration.

[0026] In another embodiment, the invention relates to the methods as described herein, wherein sleep quality is defined according to any of the group consisting of decreased sleep onset latency, decreased sleep latency, improved sleep efficiency, decreased sleep disturbance, decreased wake after sleep onset, decreased number of awakenings, improved ratio of sleep stages, decreased medication use, decreased daytime dysfunction, feeling rested / refreshed, and feeling less sleepy after waking up.

[0027] Additionally, the invention relates to methods as described herein, wherein the subject is a human.

[0028] BRIEF DESCRIPTION OF THE DRAWINGS

[0029] With specific reference now to the figures, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of the different embodiments of the present invention only. They are presented in the cause of providing what is believed to be the most useful and readily description of the principles and conceptual aspects of the invention. In this regard no attempt is made to show structural details of the invention in more detail than is necessary for a fundamental understanding of the invention. The description taken with the drawings making apparent to those skilled in the art how the several forms of the invention may be embodied in practice.

[0030] Fig. 1 : Self-assessed quality scores related to sleep (A), number of awakenings per night (B), and sleep latency (C) at baseline, after two weeks and after four weeks of supplementation. Data are expressed as estimated marginal means and error bars represent the standard error of the mean, based on a one-way repeated measure ANOVA analysis, with baseline caffeine consumption as a covariate. *: p<0.05; #: p<0.1.

[0031] Fig. 2: Stress levels of participants at baseline and after 4 weeks.

[0032] Fig. 3: Self-assessed mood scores at baseline, after two weeks and after four weeks of supplementation. Data are expressed as estimated marginal means and error bars represent the standard error of the mean, based on a one-way repeated measure ANOVA analysis. The scores are derived from a 11 -point scale ranging from a minimum of 0 “not relatable at all” to a maximum of 10 “extremely relatable”. *: p<0.05; #: p<0.1.

[0033] Fig. 4: Self-assessed sleep quality scores at baseline, after one month and after three months of supplementation. *: p<0.05; #: p<0.1 compared to baseline.

[0034] Fig. 5. Self-assessed psychological well-being scores related to emotional well-being, energy levels and perceived relaxation. *: p<0.05; #: p<0.1 compared to baseline. Fig. 6: Self-assessed skin quality scores related to skin elasticity, skin hydration, skin redness, skin itching and overall skin quality. *: p<0.05; #: p<0.1 compared to baseline.

[0035] DETAILED DESCRIPTION OF THE INVENTION

[0036] As described herein before, in a first aspect, the invention provides the use of hesperidin and naringin or hesperidin and naringenin for improving sleep quality. The term “sleep quality” as used herein refers to the following definitions: decreased sleep onset latency, decreased sleep latency, improved sleep efficiency, decreased sleep disturbance, decreased waking up after sleep onset, decreased number of awakenings, improved ratio of sleep stages, decreased medication use, decreased daytime dysfunction, feeling rested / refreshed, and feeling less sleepy after waking up.

[0037] In the context of the present invention, sleep onset latency or sleep latency is defined by the time it takes to fall asleep when trying to fall asleep. Preferably, the sleep (onset) latency is defined by the amount of minutes it takes to fall asleep after trying to fall asleep. In a preferred embodiment, the invention therefore relates to hesperidin and naringin or hesperidin and naringenin, or a composition as described herein for in use in decreasing the sleep (onset) latency, and in particular, decreasing the amount of time it takes to fall asleep after trying to fall asleep.

[0038] Further, in the context of this invention, sleep efficiency is defined by the time spent asleep divided by the total time spent in bed multiplied by 100%. The invention therefore also relates, in a preferred embodiment, to hesperidin and naringin or hesperidin and naringenin, or a composition as described herein for improving sleep efficiency. In particular, the invention relates to the use of hesperidin and naringin or hesperidin and naringenin, or a composition as described herein for improving the time spent asleep divided by the total time spent in bed multiplied by 100%.

[0039] Sleep disturbance is defined by having problems which prevent a subject from sleeping. Such problems may be waking up in the middle of the night or early morning, having to get up to use the bathroom, not able to breathe comfortably, coughing or snoring loudly, feeling too cold or too hot, having bad dreams or having pain. In a preferred embodiment, the invention relates to the use of hesperidin and naringin or hesperidin and naringenin, or a composition as described herein, for decreasing sleep disturbance.

[0040] In the context of this invention, waking up after sleep onset or number of awakenings is defined by the total amount of time awake after falling asleep and / or the number of times a subject wakes up after sleep onset. In a particular embodiment, the invention therefore relates to the use of hesperidin and naringin or hesperidin and naringenin, or the composition as described herein for decreasing waking up after sleep onset or decreasing the number of awakenings, and in particular, for decreasing the total amount of time awake after falling asleep and / or the number of times a subject wakes up after sleep onset.

[0041] Also, in the context of the present invention, the ratio of sleep stages is defined by the ratio of the light sleep stage, deep sleep stage and REM sleep stage. Therefore, the invention also relates to the use of hesperidin and naringin or hesperidin and naringenin, or the composition as described herein for improving the ratio of the light sleep stage, deep sleep stage and REM sleep stage.

[0042] Also, the invention relates to the use of hesperidin and naringin or hesperidin and naringenin, or the composition as described herein for decreasing medication use, decreased daytime dysfunction, feeling rested / refreshed, and / or feeling less sleepy after waking up.

[0043] In another specific embodiment, the invention relates to the use of hesperidin and naringin or hesperidin and naringenin, or a composition as described herein for improving mood and mental well-being. In particular, the invention relates to the use of hesperidin and naringin or hesperidin and naringenin, or a composition as described herein for reducing stress.

[0044] In another embodiment, the invention relates to hesperidin and naringin or hesperidin and naringenin or a composition as described herein for use in improving sleep quality.

[0045] In a certain production method, hesperidin is extracted from the whole fruit of sweet oranges (immature dried fruit). An extracted liquid is retrieved following alkaline extraction. This liquid is treated with sulfuric acid (H2SO4), which is eventually treated with a mixture of H2SO4 and sodium hydroxide (NaOH) to obtain the composition as defined herein.

[0046] A further embodiment provides hesperidin and naringin or hesperidin and naringenin in a composition, optionally co-extracted or combined with hesperidin in a composition for use in improving sleep quality.

[0047] In a particular embodiment, the invention relates to the composition as described herein, wherein hesperidin and naringin or hesperidin and naringenin is present in at least 0.1 wt.%, preferably in at least 1 wt.%, more preferably in at least 2 wt.%, even more preferably in between 3 and 7 wt.%, most preferably in about 5 wt.%. Further, the invention relates to the use of acomposition as described herein for improving sleep quality, wherein hesperidin is present in at least 30 wt.%, preferably in at least 50 wt.%, more preferably in at least 70 wt.%, even more preferably in between 80 and 90 wt.%, most preferably in about 84 wt.%.

[0048] In yet another particular embodiment, the invention relates to the use of a composition as described herein for use in improving sleep quality, wherein hesperidin and naringin or hesperidin and naringenin are present in a ratio hesperidimnaringin or hesperidin: naringenin of between 1:1 and 100:1, preferably in a ratio of between 1:2 and 1:50, more preferably in a ratio of between 1:10 and 1:30, most preferably in a ratio of about 1:17.

[0049] Studies have shown good bioavailability of hesperidin and naringin or hesperidin and naringenin and analogues described herein as well as hesperidin when administered orally. It is therefore an object of the invention to provide the use of compounds and compositions comprising the compounds described herein that can be orally administered, for improving sleep quality. Preferred compositions for oral administration include but are not limited to:

[0050] Sub-lingual films, tablets, or lozenges (tablets that slowly dissolve in the mouth) that ensure uptake of the compounds described herein through sublingual and / or buccal mucosa, bypassing the intestinal microflora and hepatic circulation.

[0051] Composition of citrus fruit or peel extract comprising the compounds described herein, which is active when taken orally in typical dosage forms like capsules, beverages, and food products.

[0052] In a particular embodiment, the composition is an oral composition in the form of a liquid, solution, tablet, pill, powder, lozenge, dissolvable film, suspension, dietary gel, capsule, chewable, gummy or syrup.

[0053] In a preferred embodiment, a composition of the invention comprises an isolated hesperidin and naringin or hesperidin and naringenin or the composition has been enriched for hesperidin and naringin or hesperidin and naringenin. In another particular embodiment, the composition of the invention comprises a compound of the invention and a bulking agent. The bulking agent is preferably of natural origin. Examples of bulking agents suitable for the method of administration are known to the person skilled in the art. Preferably, the bulking agent is a saccharide, including mono-, oligo- and polysaccharides; in particular a sugar or natural gum. In a further embodiment, the bulking agent is selected from the below list described for pharmaceutical carriers. The invention relates to a composition comprising a compound of the invention, wherein the composition is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier. Examples of pharmaceutical carriers are known to the person skilled in the art and include but are not limited to D-mannitol, sorbitol, sucrose, galactose, cellulose, other sugars, other saccharides, other oligosaccharides and / or polysaccharides, gelatin, guar gum, Arabic gum, agar agar, xanthan gum, locust bean gum, starch, starch fragments, dextrins, British gum and any mixture thereof. Preferably, the pharmaceutical acceptable carrier is of natural origin.

[0054] In another aspect, the invention relates to a composition comprising hesperidin and naringin or hesperidin and naringenin, wherein the composition is in a form suitable for sublingual and / or buccal administration, wherein the composition is in the form of a liquid solution, (disintegrating) tablet, pill, powder, lozenge, dissolvable film, suspension, (dietary) gel, capsule, chewable, gummy or syrup.

[0055] ‘Tablets’ refer to a dosage form comprising a mixture of one or more active substances including inner filler substances such as, disintegrant, binding agents, lubricants, glidants, anti-adherents, flavors, coloring agents etc.

[0056] ‘Disintegrating tablets’ are tablets with a specific composition that quickly disintegrates when getting in contact with saliva in the mouth or with water. These tablets may have the same composition as regular tablets, however, added with components optimized for disintegration time, e.g. by adding disintegrants.

[0057] ‘Capsules’ typically consist of a thin layer outer wall, made from a substance that dissolves in the stomach and / or intestinal fluid, e.g. gelatin, agar etc. Capsules can be filled with the active substance (i.e. hesperidin and naringin or hesperidin and naringenin ) as a dry powder, or as a fluid consisting of different solutions etc. that contain the active ingredient.

[0058] ‘Syrups’ are sweetened, flavored, viscose solutions that contain a certain concentration of the active substance, which can be diluted into a food or drink.

[0059] ‘Dietary gels’ refers to highly viscous aqueous solutions of active substances that may be added with flavoring and coloring agents in the presence of a gelling agent such as gelatin, agar, acacia gums and others. This mixture of components is presented as a slowly flowing or semi-solid substance. ‘Lozenges’ refer to hardened mixtures of the active ingredient and additional substances that slowly dissolve when getting into contact with the saliva in the mouth, elaborating a prolonged exposure of the active substance with the buccal mucosa. Lozenges often contain flavoring and sweeteners to increase palatability.

[0060] The active ingredient hesperidin and naringin or hesperidin and naringenin or a composition of the invention can be applied in any form, according to the various embodiments of the present invention. For instance, the dosage form of the active ingredient may be provided as, e.g., a powder beverage mix, beverage syrup, a liquid beverage, a ready-to-eat bar or drink product, a capsule, a liquid capsule, a tablet, a caplet, a lozenge, a chewable tablet, pill, powder, dissolvable film, a dietary gel, gummy or syrup.

[0061] Therefore, in one embodiment, the invention relates to a composition comprising hesperidin and naringin or hesperidin and naringenin or a composition of the invention, wherein the composition is a food, beverage, or supplement composition for a food composition, for example a nutrition bar or a ready to drink powdered beverage.

[0062] In one embodiment, the invention relates to a composition comprising hesperidin and naringin or hesperidin and naringenin or a composition of the invention, wherein the composition is a food, beverage, or supplement composition for a food or a beverage, for example a nutrition bar or drinks, used as medical food.

[0063] In another embodiment, the invention refers to a composition comprising hesperidin and naringin or hesperidin and naringenin or a composition of the invention, wherein the composition is in a form suitable for sublingual and / or buccal administration, wherein the composition is a food, beverage, or a supplement composition for a food or a beverage, for example a nutrition bar.

[0064] Hesperitin-7-rutinoside after extraction without enantiomeric enrichment, is a mixture of R- and S- enantiomers, the R:S molar ratio is typically between 1:1 and 1:5, depending on its source. A mixture of R- and S -enantiomers which has a molar ratio of 1:1 is also known as a racemic mixture. Hesperidin may refer to racemic hesperidin, preferably to enantiomerically enriched hesperidin, more preferably hesperidin which is enantiomerically enriched in (2S)- hesperidin.

[0065] With ‘enantiomerically enriched’ is meant that the hesperidin has an excess of one enantiomer over the other. Enantiomeric enrichment can be achieved amongst other methods known by a person skilled in the art by selection of the botanic source of hesperidin, and / or stereochemical separation techniques, such as capillary electrophoresis (CE). It has been observed that the compounds of the invention have an increased bioavailability when enantiomerically enriched.

[0066] Therefore, in a particular embodiment, the invention relates to the composition as described herein, wherein the hesperidin is enantiomerically enriched for 2S-hesperidin. In particular, the invention relates to hesperidin, or the composition as described herein, wherein at least 60%, more preferably 70%, of hesperidin is 2S-hesperidin.

[0067] The optimal dose of the compounds of the invention (formula I, la, II or Ila) for humans, preferably human adults, can be derived using the FDA recommendations for translating to the human equivalent dose (HED) from animal studies. This recommendation translates the dosages used for animal studies to dosages per day for the general population (i.e. average human adults). Preferably, the compound of the invention is administered (to a human adult) once daily in an amount of approximately 50-1500 mg. The ingredient is preferably administered during the evening.

[0068] In another particular embodiment, naringin or naringenin is present in an amount of 2.5 mg to 150 mg per unit dosage of said composition. In a particular embodiment, the naringin or naringenin is present in an amount of 3 mg to 100 mg, more particular in an amount of 5 mg to 50 mg, most particular in an amount of 10 mg to 30 mg per unit dosage of said composition. In another embodiment in about 12.5 mg or 25 mg per unit dosage of said composition, wherein naringin or naringenin is present per unit dosage in an amount as described above for daily administration.

[0069] Further, the present invention provides the daily administration of the composition of the invention comprising hesperidin in an amount of 50 mg to 1500 mg; in particular from 100 mg to 1000 mg; more in particular from 150 mg to 750 mg. In yet another particular embodiment from 150 mg to 600 mg; in particular from 200 mg to 500 mg. In another embodiment in about 210 mg or 420 mg. In a preferred embodiment, the invention provides a composition in unit dosages, wherein hesperidin or a composition as described herein is present per unit dosage in an amount as described above for daily administration. In a further embodiment, hesperidin is present in an amount of 50 mg to 1500 mg per unit dosage of said composition.

[0070] In another embodiment, the invention relates to a method for improving sleep quality in a subject comprising a step of administering hesperidin and naringin or hesperidin and naringenin or a composition as described herein to a subject. In a more specific embodiment, the invention relates to this method, wherein the hesperidin and naringin or hesperidin and naringenin or the composition is administered via oral administration.

[0071] Additionally, the invention relates to these methods, wherein sleep quality is defined according to any of the group consisting of decreased sleep onset latency, decreased sleep latency, improved sleep efficiency, decreased sleep disturbance, decreased wake after sleep onset, decreased number of awakenings, improved ratio of sleep stages, decreased medication use, decreased daytime dysfunction, feeling rested / refreshed, and feeling less sleepy after waking up.

[0072] The subject to be treated with the hesperidin and naringin or hesperidin and naringenin, or the composition of the invention, and / or by the method of the invention is preferably a mammal, in particular a human.

[0073] Without being bound by theory, reduced sleep quality can result in stress and stress can negatively affect skin health. The reduced sleep quality can therefore result in increased skin inflammation, increased itching, impaired skin barrier function, impaired wound healing, suppressed immunity. Enhancing sleep quality therefore also results in improving sleep health. Subject having enhanced sleep quality, therefore show less skin aging signs, better skin barrier function, better recovery after UV exposure and decreased skin inflammation. Skin inflammation may cause outbreaks of acne, eczema, psoriasis, skin allergies.

[0074] In the context of this invention, improving skin health can be defined as enhancing skin elasticity and firmness, reducing wrinkle depth, reducing trans epidermal water loss, increasing skin hydration, increasing skin moisturization, increasing skin homogeneity, increasing skin sebum content, reducing skin cellular oxidative stress, reducing skin inflammation, reducing skin redness, reducing outbreaks of acne, eczema, psoriasis, skin allergies, enhancing skin microbiome composition.

[0075] Thus, in another embodiment, the present invention also relates to the use of hesperidin and naringin or hesperidin and naringenin for improving skin health, wherein hesperidin and naringin or hesperidin and naringenin are administered via oral administration. The invention also relates to the use of hesperidin and naringin or hesperidin and naringenin for enhancing skin elasticity and firmness, reducing wrinkle depth, reducing trans epidermal water loss, increasing skin hydration, increasing skin moisturization, increasing skin homogeneity, increasing skin sebum content, reducing skin cellular oxidative stress, reducing skin inflammation, reducing skin redness, reducing outbreaks of acne, eczema, psoriasis, skin allergies, enhancing skin microbiome composition, wherein hesperidin and naringin or hesperidin and naringenin are administered via oral administration.

[0076] Therefore, in a specific embodiment, the invention relates to the use of hesperidin and naringin or hesperidin and naringenin in improving sleep quality, wherein hesperidin and naringin or hesperidin and naringenin are administered via oral administration, and wherein the improved sleep quality leads to improved skin health.

[0077] In a more specific embodiment, the invention also relates to a method for improving sleep quality in a subject comprising a step of orally administering hesperidin and naringin or hesperidin and naringenin or a composition as defined herein to a subject, wherein the improved sleep quality leads to improved skin health.

[0078] EXAMPLES

[0079] Example 1

[0080] Study population

[0081] In this study, healthy men and women (aged 40 to 70 years), with a BMI between 18.5 - 30 kg / m2with (minor) sleep disturbance were included. Main exclusion criteria included: excessive caffeine use; major psychiatric / mental health disorders; chronic sleep disorders; other clear causes for poor sleep quality, stress, depression or anxiety; use of medication or supplements that can affect outcomes.

[0082] Study design

[0083] The study was designed as a randomized, double-blind, placebo-controlled, cross-over trial. Each subject underwent two different intervention periods, during which daily citrus extract or placebo was supplemented for eight weeks, with a 4-week wash-out period in between. The order of intervention was decided by a randomization procedure.

[0084] Each test day, participants arrived fasted in the morning at the test facility and a sleep diary was handed in. Then, anthropometric measurements were performed. Blood samples were collected from an antecubital vein in the forearm and the Pittsburg Sleep Quality Index (PSQI) questionnaire and other questionnaires regarding depression, anxiety and stress were filled out.

[0085] During the pre-measurements participants received capsules containing either the citrus extract or the placebo in a randomized order for eight weeks. In addition to the measurements preformed during every test day, collected feces, morning saliva and the Fitbit were handed in during the pre- and 8-week measurements.

[0086] Intervention

[0087] The investigational product in this intervention study was a citrus extract (MicrobiomeX®) containing > 80% hesperidin and > 5% naringin. The extract was provided in capsules of 250 mg. Participants were instructed to ingest two capsules in the evening during dinner with 200 mL of water. The placebo used in this study was maltodextrin.

[0088] Study parameters / endpoints

[0089] Sleep:

[0090] Sleep quality was assessed using the validated PSQI, a sleep diary (consensus sleep diary) and actigraphy measurements using the Fitbit. The PSQI is a self-reporting questionnaire that assesses sleep quality based on the last four weeks. The questionnaire contains seven components that measure subjective sleep quality, sleep latency, sleep duration, sleep efficiency, sleep disturbance, use of sleep medication, and daytime dysfunction. The general outcome is a score in the range of 0-21. A higher score is associated with decreased sleep quality, and a score > 5 indicates disturbed sleep quality.

[0091] With the consensus sleep diary, sleep onset latency, total sleep time, number of awakenings, wake after sleep onset (min), sleep efficiency, sleep quality, morning freshness and morning sleepiness were measured.

[0092] Total sleep time, number of awakenings, wake after sleep onset (min) and sleep efficiency were also measured using actigraphy. In addition, light sleep, deep sleep and REM sleep were assessed with actigraphy.

[0093] Psychological wellbeing:

[0094] Depression: depression scores were assessed using the 21-item Beck Depression Inventory (BDI).

[0095] Stress: stress was assessed using the 10-item Perceived Stress Scale (PSS) and by measurement of morning salivary cortisol.

[0096] Anxiety: anxiety scores were assessed using the 21-item Beck Anxiety Inventory (BAI).

[0097] Biomarkers related to mental well-being:

[0098] On each test day, blood samples were collected from an antecubital vein in the forearm. Serum and plasma concentrations of BDNF were assessed by ELISA. In addition, measurements of serotonin and dopamine concentrations were performed.

[0099] Example 2

[0100] Study population

[0101] Participants were aged 35-70 years and in general good health. Subjects were excluded when they had sleeping disturbances (such as insomnia, resting leg syndrome, shift work), mental health disorders, when they were in perimenopause, allergic to any of the product substances, or when they used sleep medication or consumed oranges or orange juice on a daily basis. When subjects had travelled from a place with >3 -hour time difference in < 1 weeks, they had to wait to participate until at least 1 week had passed. In addition, average daily caffeine consumption was estimated at baseline. Study design

[0102] Once included, participants were instructed to maintain their habitual diet. Before the start of the intervention, participants received a logbook, sleep diary and the capsules. Online questionnaires assessing sleep, stress and mental health were filled in three times by the participants; before supplementation, after two weeks and after four weeks of supplementation. Participants were instructed to fill in a sleep diary three times for 7 days; one week before the supplementation period, after one week of supplementation and after three weeks of supplementation. After completion of the study, participants returned the sleep diary together with the logbook.

[0103] Intervention

[0104] Participants had to consume 2 capsules, each containing 250 mg MicrobiomeX® (citrus extract containing >80% hesperidin and >5% naringin) and 100 mg maltodextrin, per day during dinner, for 4 weeks.

[0105] Study parameters / endpoints

[0106] The consensus sleep diary Core was used to assess the main parameters of sleep during a period of 7 days, before baseline, before the 2-week measurements and before the 4-week measurements. Additional questions regarding sleep quality and dream quality were asked in the online questionnaire at baseline, after two weeks and after four weeks. At baseline and in the 4-week questionnaire the validated Perceived Stress Scale (PSS) was used to examine participants’ stress levels. The baseline scores were used to categorize subjects in low, middle, and high stress groups. Additional questions in regards to mood were asked. Compliance was checked by asking how many capsules were left in the jar at the last day of supplementation and by checking the logbooks. If more than 80% of the required capsules were consumed, participants were considered compliant and included in the analysis.

[0107] Scores for sleep onset latency, number of awakenings during the night and sleep quality of each of the three sleep diaries were averaged and used for further analysis. If participants had special reasons why sleep was disturbed during a night (e.g., illness, noise outside), these nights were excluded from the calculation. The total PSS scores were calculated by scoring each question 0-4 (never = 0, very often = 4), the scores of positively formulated questions were reversed. Total scores 0 - 13, 14-26 and 27-40 were considered as low, middle and high stress scores, respectively.

[0108] Results

[0109] Baseline characteristics Baseline characteristics of the study population are shown in Table 1. In total, 19 participants were eligible to participate, of which 17 participants completed the study.

[0110] Characteristics Total (17)

[0111] Sex (male / female n (% male)) 4 / 13 (23.5%) 5 Age (years, mean + st. dev.) 56.0 + 7.0 BMI (kg / m2, mean + st. dev.) 25.5 + 2.6 Daily caffeine consumption 294 + 155 (mg, mean + st. dev.)

[0112] Table 1 Baseline characteristics of the study population.

[0113] Sleep related outcomes

[0114] Mean scores for sleep quality increased and the number of awakenings during the night significantly decreased after two weeks and after four weeks of supplementation, compared to baseline (all p<0.001; Figure 1 A,B).

[0115] Daytime sleepiness scores decreased significantly at two weeks, compared to baseline (p=0.026). In addition, after 4 weeks of supplementation there was a significant decrease in sleep latency (p=0.009; Figure 1 C), which is the time it takes a person to fall asleep after trying to fall asleep. Furthermore, an upward trend for dream quality and a downward trend for sleepiness after waking up were visible after two and after four weeks, respectively (both p< 0.073).

[0116] Perceived stress scores

[0117] Perceived stress scores were measured at baseline and after four weeks of supplementation. Based on the Perceived Stress scale questionnaire, participants were categorized into low, middle and high stress groups at baseline (Figure 2). After four weeks, the average stress score had decreased from 15.8 to 13.4, and 22% of the participants that were categorized at baseline as middle stressed, were categorized as low stressed after four weeks. The participant that was highly stressed, had moved to the middle-stressed group.

[0118] Mood related outcomes

[0119] To assess whether MicrobiomeX® supplementation improved mental wellbeing, different factors related to mood were assessed. The questions were rated 0-10, based on how relatable the statements were. Supplementation with citrus extract resulted in improved scores for ‘encouraged’ and ‘active’. For both factors, these changes were significant after 2 weeks of supplementation (both p< 0.032; Figure 3) and remained increased after 4 weeks of supplementation compared to baseline (both p< 0.098).

[0120] Example 3

[0121] Methods

[0122] The study was designed to determine the effect of citrus flavonoids on outcomes of sleep quality, mental well-being and skin quality. The participants consisted of healthy individuals and those reporting skin issues like eczema and dermatitis.

[0123] Each participant received capsules containing 500 mg of citrus extract (MicrobiomeX®; > 80% hesperidin and > 5% naringin) for a duration of three months. Capsules were ingested daily for the duration of the study.

[0124] After inclusion, a baseline measurement took place. After baseline measurements were completed, participants ingested the study product for a duration of three months, with a measurement after one month of intake and a measurement after three months of intake. During these measurements, participants are asked to complete a questionnaire to assess self-perceived sleep quality, psychological well-being and skin quality. Perceived quality during the past week was scored on an 11-point Likert scale ranging from a minimum of 0 “very low / poor” to a maximum of 10 “very high / good”. Emotional wellbeing was assessed by asking participant how happy or content they felt, and perceived relaxation by scoring how relaxed or free of tension participants they felt during the past week.

[0125] Results

[0126] In total, data from 56 participants who completed all three questionnaires were included in the analyses.

[0127] Sleep quality

[0128] Overall sleep quality scores showed a significant improvement from baseline to one month following the initiation of supplementation, with a further significant enhancement observed after three months of supplementation from baseline (all p<0.001; Figure 4).

[0129] Psychological well-being

[0130] Psychological well-being and energy scores are displayed in Figure 5. The results for emotional well-being, energy levels and perceived relaxation show a significant improvement from baseline following one month of supplementation (all p<0.001) and these scores remained increased throughout the three-month assessment. This shows that participants indicated that they felt more happy, having more energy and more relaxed and free of tension after intake of the citrus extract. Skin quality

[0131] Skin quality related outcomes are displayed in Figure 6. Skin hydration, skin itching, skin elasticity and overall skin quality scores demonstrated significant improvement from baseline to one month following the initiation of supplementation (all p<0.015) and remained stable through the three- month period (p<0.001). Furthermore, an overall decrease in skin redness scores was reported (p=0.056). These results show that the improved sleep quality scores resulted in an improvement in skin health outcomes.

Claims

CLAIMS1. Use of hesperidin and naringin or hesperidin naringenin for improving sleep quality, wherein hesperidin and naringin or hesperidin and naringenin is administered via oral administration.

2. Use as claimed in claim 1, wherein the composition further comprises isonaringin, neoponcirin and / or hesperitin.

3. Use as claimed in any one of claims 1 or 2, wherein naringin or naringenin is present in at least 0.1 wt.%, preferably in at least 1 wt.%, more preferably in at least 2 wt.%, even more preferably in between 3 and 7 wt.%, most preferably in about 5 wt.% of the composition as used in claim 1.

4. Use as claimed in any one of claims 1-3, wherein hesperidin is present in at least 30 wt.%, preferably in at least 50 wt.%, more preferably in at least 70 wt.%, even more preferably in between 80 and 90 wt.%, most preferably in about 84 wt.% of the composition as used in claim 1.

5. Use as claimed in any one of claims 1-4, wherein the composition is a pharmaceutical composition, optionally comprising an excipient or pharmaceutically acceptable carrier.

6. Use as claimed in any one of claims 1-5, wherein the use is a non-therapeutic use.

7. Use as claimed in any one of claims 1-6, wherein said composition is in the form of a liquid, solution, tablet, pill, powder, lozenge, dissolvable film, suspension, dietary gel, capsule, chewable, gummy or syrup.

8. Use as claimed in any one of claims 1-7, wherein hesperidin is enantiomerically enriched for 2S-hesperidin.

9. Use as claimed in any one of claims 1-8, wherein at least 60%, more preferably 70%, of hesperidin of the composition used is 2S-Hesperidin.

10. Use as claimed in any one of claimsl-9, wherein the sleep quality is defined according to any of the group consisting of: decreased sleep onset latency, decreased sleep latency, improved sleep efficiency, decreased sleep disturbance, decreased wake after sleep onset, decreased numberof awakenings, improved ratio of sleep stages, decreased medication use, decreased daytime dysfunction, feeling rested / refreshed, and feeling less sleepy after waking up.

11. Use claimed in any one of claims 1-10, wherein the improved sleep quality leads to improved skin health.

12. Method for improving sleep quality in a subject comprising a step of orally administering hesperidin and naringin or hesperidin and naringenin or a composition as defined in any one of claims 1-11 to a subject.

13. Method as claimed in claim 13, wherein sleep quality is defined according to any of the group consisting of: decreased sleep onset latency, decreased sleep latency, improved sleep efficiency, decreased sleep disturbance, decreased wake after sleep onset, decreased number of awakenings, improved ratio of sleep stages, decreased medication use, decreased daytime dysfunction, feeling rested / refreshed, and feeling less sleepy after waking up.

14. A method as claimed in any one of claims 12 or 13, wherein the improved sleep quality leads to improved skin health.

15. Method as claimed in any one of claims 12 or 13, wherein the subject is a human.

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