WO2022115907A1

WO2022115907A1 - Methods for improving sleep quality

Info

Publication number: WO2022115907A1
Application number: PCT/AU2021/051432
Authority: WO
Inventors: Wayne FINLAYSON
Original assignee: Servatus Ltd
Priority date: 2020-12-01
Filing date: 2021-12-01
Publication date: 2022-06-09
Also published as: EP4255456A1; EP4255456A4

Abstract

Provided herein are methods for inducing or maintaining sleep, for improving sleep quality, or for treating insomnia, in human subjects, comprising administering an effective amount of Lactobacillus plantarum and/or Lactobacillus zeae, or a culture supernatant or cell free filtrate derived from culture media in which said Lactobacillus species has been cultured. In an embodiment, the method comprises administering a microbial biotherapeutic composition comprising Lactobacillus plantarum and Lactobacillus zeae.

Description

Methods for improving sleep quality

Field of the Art

[0001] The present disclosure relates generally to methods to aid in the induction and/or maintenance of sleep in a subject, and for improving the quality of sleep in a subject, including the treatment of insomnia, comprising the administration of compositions comprising one or more microorganisms, or culture supernatants or cell free filtrates derived from culture media in which the one or more microorganisms has been cultured.

Background

[0002] Sleep is clinically important for many reasons. Myriad studies have demonstrated associations between length and quality of sleep and cognitive performance (such as learning and memory processes), mental health, general health and wellbeing. Inadequate or disrupted sleep reduces necessary overnight brain activity that is needed for higher order neurocognitive functioning, and results in a decline in the ability to complete complex tasks requiring abstract thinking, creativity, memory and planning. Sustained patterns of poor or inadequate sleep are also associated with poor modulation of immunity, a reduction in natural immune responses and T cell cytokine production, and an adverse effect on resistance to infectious disease. Significant associations have also been observed between impaired sleep and hypertension. Poor sleep patterns have a negative effect on quality of life and are risk factors for the development of numerous physiological insults and chronic conditions including oxidative stress, inflammation, depression, obesity, metabolic syndrome, cardiovascular disease, and cancer.

[0003] Sleep disturbance, including insomnia, is increasingly common. Epidemiological surveys have indicated that up to about 40% of the adult population (often more than 50% in older adults) complain of impaired sleep quality on a regular basis. Typical complaints surround a difficulty or inability to fall asleep, waking up repeatedly and difficulty or an inability to go back to sleep, and difficulty getting enough sleep. Broadly speaking, four types of insomnia are generally recognised: sleep onset insomnia; sleep maintenance insomnia; early morning awakening; and transient insomnia.

[0004] Many treatments and aids have been devised over the years to combat a lack of sleep and/or poor sleep quality, including for example, breathing machines, eye masks, behavioural modifications and drugs such as sedating antihistamines, herbal extracts, sedating antidepressants and benzodiazepines, the latter at least being associated with some serious side effects such as addiction.

[0005] Notwithstanding a growth in the number of type of medications and other sleep aids available, sleep disorders and complaints of lack of sleep and/or poor quality of sleep are becoming more common. There clearly remains a need for improved methods for improving sleep.

Summary of the Disclosure

[0006] One aspect of the present disclosure provides a method for inducing or maintaining sleep in a human subject, comprising administering to the subject an effective amount of Lactobacillus plantarum and/or Lactobacillus z.eae. or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured.

[0007] In a particular embodiment, the method comprises administering a composition comprising Lactobacillus plantarum. In another embodiment, the method comprises administering a composition comprising Lactobacillus .eae.

[0008] In a particular embodiment, the method comprises administering an effective amount of Lactobacillus plantarum and Lactobacillus zeae. Typically, the Lactobacillus plantarum and Lactobacillus zeae are present in the same composition.

[0009] The method may further comprise administering one or more additional agents, such as one or more Lactobacillus species optionally selected from Lactobacillus buchneri, Lactobacillus rapi, Lactobacillus paracasei, Lactobacillus parafarraginis, and Lactobacillus diolivorans, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured.

[00010] The method may comprise administering a composition comprising Lactobacillus plantarum and Lactobacillus zeae in the form of a liquid or solid unit dosage form, a food or a beverage.

[00011] Another aspect of the present disclosure provides a method for improving sleep quality in a human subject, comprising administering to the subject an effective amount of Lactobacillus plantarum and/or Lactobacillus zeae, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured.

[00012] Improving sleep quality may comprise improving one or more of the onset of sleep, sleep depth, sleep duration, or fatigue and/or lethargy following sleep. [00013] In a particular embodiment, the method comprises administering a composition comprising Lactobacillus plantarum. In another embodiment, the method comprises administering a composition comprising Lactobacillus zeae.

[00014] In a particular embodiment, the method comprises administering an effective amount of Lactobacillus plantarum and Lactobacillus zeae. Typically, the Lactobacillus plantarum and Lactobacillus zeae are present in the same composition.

[00015] The method may comprise administering a composition comprising Lactobacillus plantarum and Lactobacillus zeae in the form of a liquid or solid unit dosage form, a food or a beverage.

[00016] The method may further comprise administering one or more additional agents, such as one or more Lactobacillus species optionally selected from Lactobacillus buchneri, Lactobacillus rapi, Lactobacillus paracasei, Lactobacillus parafarraginis, and Lactobacillus diolivorans, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured.

[00017] Another aspect of the present disclosure provides a method for treating insomnia in a human subject, comprising administering to the subject an effective amount of Lactobacillus plantarum and/or Lactobacillus zeae, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured.

[00018] The insomnia may be sleep onset insomnia, sleep maintenance insomnia, early morning awakening, or transient insomnia.

[00019] In a particular embodiment, the method comprises administering a composition comprising Lactobacillus plantarum. In another embodiment, the method comprises administering a composition comprising Lactobacillus zeae.

[00020] In a particular embodiment, the method comprises administering an effective amount of Lactobacillus plantarum and Lactobacillus zeae. Typically, the Lactobacillus plantarum and Lactobacillus zeae are present in the same composition.

[00021] The method may further comprise administering one or more additional agents, such as one or more Lactobacillus species optionally selected from Lactobacillus buchneri, Lactobacillus rapi, Lactobacillus paracasei, Lactobacillus parafarraginis, and Lactobacillus diolivorans, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured. [00022] The method may comprise administering a composition comprising Lactobacillus plantarum and Lactobacillus zeae in the form of a liquid or solid unit dosage form, a food or a beverage.

[00023] Another aspect of the present disclosure provides a composition comprising Lactobacillus plantarum and/or Lactobacillus zeae, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured, when used for inducing or maintaining sleep, for improving sleep quality, and/or for treating insomnia.

[00024] In accordance with aspects and embodiments of the present disclosure, the method may comprise administering to the human subject a microbial biotherapeutic composition comprising Lactobacillus plantarum and/or Lactobacillus zeae. In a particular embodiment the microbial biotherapeutic composition comprises Lactobacillus plantarum and Lactobacillus zeae. The microbial biotherapeutic composition may be administered in the form of, for example, a liquid or solid unit dosage form, a food or a beverage.

[00025] Other aspects of the present disclosure comprise methods for inducing or maintaining sleep, for improving sleep quality, and/or for treating insomnia in a human subject comprising administering to the subject an effective amount of Lactobacillus zeae, Lactobacillus paracasei and Lactobacillus parafarraginis, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species have been cultured.

[00026] The method may comprise administering to the human subject a microbial biotherapeutic composition comprising Lactobacillus zeae, Lactobacillus paracasei and Lactobacillus parafarraginis.

Brief Description of the Drawings

[00027] Embodiments of the present disclosure are described herein, by way of non-limiting example only, with reference to the following figures:

[00028] Figure 1. Weekly mean Pittsburgh Sleep Quality Index (PSQI) score (± SEM) from subjects administered L. zeae only (circles), L. plantarum only (triangles) or L. zeae and L. plantarum (squares) over five weeks. Significance values (p-values) versus baseline are shown below the graph.

[00029] Figure 2. Mean change in weekly PSQI score (± SEM) from subjects administered L. zeae only (circles), L. plantarum only (triangles) or L. zeae and L. plantarum (squares) over five weeks. Dotted line represents Clinically Meaningful Improvement (CMI). Significance values (p-values) versus baseline are shown below the graph. [00030] Figure 3. Overall mean PSQI scores from baseline (± SEM) (squares) at weeks

0 and 5 for (from left to right) L. zeae only, L. zeae and L. plantarum, and L. plantarum only. Significance values (p-values) versus baseline are shown below the graph.

[00031] Figure 4. Weekly mean general wellbeing score (± SEM) from subjects administered L. zeae only (circles), L. plantarum only (triangles) or L. zeae and L. plantarum (squares) over five weeks. Significance values (p-values) versus baseline are shown below the graph.

[00032] Figure 5. Weekly mean change in general wellbeing score (± SEM) from subjects administered L. zeae only (circles), L. plantarum only (triangles) or L. zeae and L. plantarum (squares) over five weeks. Significance values (p-values) versus baseline are shown below the graph.

[00033] Figure 6. Number of responders in three treatment groups based on a clinically meaningful improvement (CMI) based on PSQI score measured weekly (> 3-point reduction in PSQI score). Weeks 1 to 5 shown from left to right for each treatment group.

Detailed Description

[00034] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, typical methods and materials are described.

[00035] The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[00036] In the context of this specification, the term "about," is understood to refer to a range of numbers that a person of skill in the art would consider equivalent to the recited value in the context of achieving the same function or result.

[00037] Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[00038] As used herein the term "effective amount” includes within its meaning a non-toxic but sufficient amount of a composition to provide the desired effect and/or physiological response. The exact amount required will vary from subject to subject depending on factors such as the species being treated, the age and general condition of the subject, the severity of the condition being treated, the particular agent being administered and the mode of administration and so forth. For any given case, an appropriate “effective amount” may be determined by one of ordinary skill in the art using only routine experimentation.

[00039] As used herein the terms "treating", “treatment” and the like refer to any and all applications which remedy, or otherwise hinder or retard insomnia. Thus, treatment does not necessarily imply that a subject is treated until complete resolution of, or recovery from, insomnia.

[00040] The term "optionally" is used herein to mean that the subsequently described feature may or may not be present or that the subsequently described event or circumstance may or may not occur. Hence the specification will be understood to include and encompass embodiments in which the feature is present and embodiments in which the feature is not present, and embodiments in which the event or circumstance occurs as well as embodiments in which it does not.

[00041] In the context of this specification, the term “microbial biotherapeutic” is to be given its broadest construction and is understood to refer to a microbial cell population or preparation, or component of a microbial cell population or preparation, which when administered to a subject in an effective amount promotes a health benefit in the subject. The cell population may comprise bacteria from one species or strain, or alternatively from multiple species or strains.

[00042] In the context of this specification, the term “prebiotic” is to be given its broadest construction and is understood to refer to any non-digestible substance that stimulates the growth and/or activity of commensal beneficial bacteria in the digestive system.

[00043] In the context of this specification, the terms "food" and "beverage" include but are not limited to health foods and beverages, functional foods and beverages, and foods and beverages for specified health use. When such foods or beverages of the present invention are used for subjects other than humans, the terms can be used to include a feedstuff.

[00044] Provided herein are methods for inducing or maintaining sleep, for improving sleep quality and for treating insomnia, wherein a human subject is administered an effective amount of a composition comprising Lactobacillus plantarum and/or Lactobacillus ~.eae. or a culture supernatant or cell free filtrate derived from culture media in which said Lactobacillus species has been cultured. [00045] Also provided are methods for inducing or maintaining sleep, for improving sleep quality, and for treating insomnia in a human subject comprising administering to the subject an effective amount of Lactobacillus zeae, Lactobacillus paracasei and Lactobacillus parafarraginis, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species have been cultured,

[00046] Subjects in need of improvement of sleep quality may exhibit perturbed sleep patterns that deviate from "normal" sleep patterns in one or more parameters, may suffer from or be at risk of perturbed sleep patterns, or may be diagnosed with a sleep disorder. In specific embodiments, the subject may suffer from an inability to fall asleep and/or from mid-sleep period awakenings. The methods of the present disclosure improve the ability of subjects to get to sleep (the onset of sleep), the ability of subjects to stay asleep (reduced mid-sleep awakenings), and improve sleep depth, sleep duration, or fatigue, lethargy or sleepiness following sleep.

[00047] The subject may suffer from insomnia or be at risk of insomnia. Insomnia and similar sleep disorders may be diagnosed by clinical interview with a psychologist, and/or may include the completion of self-reporting questionnaires and sleep diaries. Self-reporting questionnaires are employed in diagnostic tools such as the Insomnia Severity Index, the Pittsburgh Sleep Quality Index, the Insomnia Symptom Questionnaire, and the Athens Insomnia Scale.

[00048] The methods may improve the sleep pattern of a subject (sleep architecture). As used herein, "sleep pattern" refers, for a given individual, to the time spent in each of the sleep stages, including the relative proportion of the duration of each stage as compared to the duration of other stages, as measured, for example, by spectral analysis of electroencephalograms. Parameters of "normal" sleep patterns for specific populations are well known in the art (see, for example, Latta et al., 2005, Sleep 28:1525-1534). In mammals, sleep is typically divided into two broad types, rapid eye movement (REM) and non-rapid eye movement (non-REM) sleep. Non-REM can be further divided into three stages, Nl, N2, and N3 (or delta or slow-wave sleep). Sleep proceeds in cycles of REM and Non-REM. There is a greater amount of deep sleep (stage N3) earlier in the sleep cycle, while the proportion of REM sleep increases later in the sleep cycle and just before natural awakening. In humans, each sleep cycle lasts on average from 90 to 110 minutes. REM sleep, accounts for 20-25% of total sleep time in most adults. The criteria for REM sleep include rapid eye movements as well as a rapid low- voltage EEG. The methods of the present disclosure may increase the amount, frequency and/or duration of REM sleep. The methods of the present disclosure may be considered to improved sleep quality by determination of, for example, normalisation of distribution of slow-wave or REM sleep stages. [00049] In accordance with the present disclosure, assessment of sleep and sleep quality may be achieved in various ways well known to those skilled in the art. For example, a subject may be monitored overnight using electrodes and an electroencephalograms (EEGs), or may wear a device designed to monitor and measure various physiological parameters. Alternatively, a subject may complete a questionnaire assessing various aspects of lifestyle, behaviour, environment and sleep patterns or perform a self-assessment of nightly quality of sleep, in a journal for example. Suitable questionnaires include the Pittsburgh Sleep Quality Index (Buysse et al, 1989, Psychiatry Research 28: 193-213), the Insomnia Severity Index (Bastien et al., 2001, Sleep Med 2:297-307), the Insomnia Symptom Questionnaire (Spielman etal., 1987, Sleep 10:45- 56), and the Athens Insomnia Scale (Soldatos et al., 2000, J Psychosom Res 48:555-560). Examples of sleep quality assessment and determination methods are described, for example, in Mollayeva et al, 2016, Sleep Med Rev 25:52-73; Ibanez et al, 2018, PeerJ 6:e4849; Mendonca et al, 2019, IEEE Access 7:24527-24546. During or following treatment in accordance with the present disclosure, such questionnaires may be completed, for example daily or weekly and results may be compared to results from the same questionnaires completed prior to initiation of the treatment to determine improvements in sleep quality.

[00050] Improvements in sleep quality may also be determined objectively using a number of tools, including Somfit measurement outputs Sleep Onset Latency (SOL), Wake After Sleep Onset (WASO), Total Sleep Time (TST), Sleep Efficiency (SE), number of awakenings (nWAK) and number of awakenings per hour of sleep (WAKI). During or following treatment in accordance with the present disclosure, such measurements may be made, for example, daily and average measures for a week may be compared to average measure prior to initiation of the treatment to determine improvements in sleep quality.

[00051] Sleep quality may be also be assessed, and hence improvements in sleep quality from methods of the present disclosure may be determined, by a variety of other measures including, for example, a change in the rate of arousal events (e.g. in total or hourly during a sleep period), number of nights requiring rescue medicine to support sleep, improvements in daytime functioning (e.g. assessed by the Work and Social Adjustment Scale (WSAS; Mundt et al., 2002, Br J Psychiatry 180:461-464)), improvements in various quality of life measures (such as the 36- Item Short Form Survey, SF-36), reductions in daytime sleepiness (e.g. assessed by the Epworth Sleepiness Scale (ESS; Johns, 1991, Sleep 14:50-55).

[00052] Those skilled in the art will appreciate that the scope of the present disclosure is not limited by reference to any specific means of assessing or determining a subject’s sleep quality or sleep patterns. [00053] In some embodiments the methods of the present disclosure comprise administering an effective amount of a composition as described herein in one or more doses per day, optionally one or two doses per day. For example the composition may be administered in a single dose before bed, or in two doses spaced apart through the day. For example, a first dose may be taken in the morning, middle of the day, mid afternoon or about 6 hours, 5 hours, 4 hours, 3 hours, 2 hours or 1 hour before bed, and a second dose may be taken about 30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours or 10 hours after the first dose. Where only a single dose is consumed per day, the dose may be taken, for example, about 15 minutes, 30 minutes, 60 minutes, 90 minutes, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours or 10 hours before bed. In some embodiments doses of the composition may also be administered, as needed, through the normal sleep period of the subject, for example if the example awakens during the normal sleep period, and optionally if the subject has difficulty getting back to sleep after such an awakening.

[00054] Typically administration of compositions in accordance with methods of the present disclosure is continued at least until sleep quality has improved. Administration may be continued, for example, for about 7 days, 10 days, 14 days, 21 days, four weeks, five weeks, six weeks, seven weeks, two months, three months, four months, five months, six months or longer. The dosing regimen may be modified or altered during the course of administration as needed by the subject or on the advice of a medical practitioner or other health care professional.

[00055] Methods of the present disclosure contemplate the administration of Lactobacillus plantarum and/or Lactobacillus z.eae. or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured. Methods of the present disclosure also contemplate the administration of Lactobacillus zeae, Lactobacillus paracasei and Lactobacillus parafarraginis, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured. In view of some taxonomic discrepancies and uncertainties, L. zeae may also be referred to elsewhere as L. casei. However this is not settled, and L. zeae can be regarded as distinct (see http://lactotax.embl.de/wuyts/lactotax/). For the purposes of the present disclosure the L. zeae nomenclature is retained.

[00056] In some embodiments the methods of the present disclosure contemplate the administration of Lactobacillus plantarum and Lactobacillus zeae, optionally in the same or different compositions. In some embodiments the methods of the present disclosure contemplate the administration of Lactobacillus zeae, Lactobacillus paracasei and Lactobacillus parafarraginis, optionally in the same or different compositions. [00057] The Lactobacillus plantarum may be L. plantarum SVT 09P1, deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM), Federal Public Planning Service Science Policy, 8 rue de la Science B-1000, Brussels, Belgium, on 10 August 2020 under Accession Number LMG P-31923.

[00058] The Lactobacillus zeae may be L. zeae SVT 08Z1, deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM) on 27 February 2019 under Accession Number LMG P-31295, and previously described in WO 2020/073088.

[00059] In the following discussion, in the context of administration of the Lactobacillus species or culture supernatants or cell free filtrates derived from culture media in which the Lactobacillus has been cultured, and in the context of compositions comprising the same, the term “Lactobacillus” may be used hereinafter to refer not only to the specific Lactobacillus species defined herein per se, but also more broadly to refer to culture supernatants or cell free filtrates derived from culture media in which the specific Lactobacillus species defined herein have been cultured. In embodiments in which culture supernatants or cell free filtrates derived from culture media in which said Lactobacillus have been cultured are employed, the bacteria may be cultured together or separately.

[00060] Methods of the present disclosure may further comprise the administration of one or more of Lactobacillus buchneri, Lactobacillus rapi, Lactobacillus paracasei, Lactobacillus parafarraginis, and Lactobacillus diolivorans, or culture supernatants or cell free filtrates derived from culture media in which said Lactobacillus have been cultured. In such embodiments the bacteria may be cultured together or separately.

[00061] The Lactobacillus parafarraginis may be L. parafarraginis SVT- 18 (which may be elsewhere referred to by the alternate designation SVT 05P2) deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM) on 27 February 2019 under Accession Number LMG P-31292.

[00062] The Lactobacillus buchneri may be L. buchneri SVT-23 (which may be elsewhere referred to by the alternate designation SVT 06B1) deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM) on 27 February 2019 under Accession Number LMG P-31293.

[00063] The Lactobacillus rapi may be L. rapi SVT-24 (which may be elsewhere referred to by the alternate designation SVT 07R1) deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM) on 27 February 2019 under Accession Number LMG P-31294. [00064] The Lactobacillus paracasei may be L. paracasei SVT-09 (which may be elsewhere referred to by the alternate designation SVT 04P1) deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM) on 27 February 2019 under Accession Number LMG P-31290.

[00065] The Lactobacillus diolivorans may be L. diolivorans SVT-03 (which may be elsewhere referred to by the alternate designation SVT 01D1) deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM) on 27 February 2019 under Accession Number LMG P-31287.

[00066] The concentrations of individual Lactobacillus species to be administered in accordance with methods of the present disclosure will depend on a variety of factors including the identity and number of individual species employed, the health and wellbeing of the subject, the nature and extent of sleep disturbance to be treated and the form and route in which a composition is administered For any given case, appropriate concentrations may be determined by one of ordinary skill in the art using only routine experimentation. By way of example only, the concentration of the Lactobacillus species, or each species present in the case of a combination, may be from about 1 x 10² cfu/ml to about 1 x 10¹¹ cfu/ml, and may be about 1 x 10³ cfu/ml, about 2.5 x 10³ cfu/ml, about 5 x 10³ cfu/ml, 1 x 10⁴ cfu/ml, about 2.5 x 10⁴ cfu/ml, about 5 x 10⁴ cfu/ml, 1 x 10⁵ cfu/ml, about 2.5 x 10⁵ cfu/ml, about 5 x 10⁵ cfu/ml, 1 x 10⁶ cfu/ml, about 2.5 x 10⁶ cfu/ml, about 5 x 10⁶ cfu/ml, 1 x 10⁷ cfu/ml, about 2.5 x 10⁷ cfu/ml, about 5 x 10⁷ cfu/ml, 1 x 10⁸ cfu/ml, about 2.5 x 10⁸ cfu/ml, about 5 x 10⁸ cfu/ml, 1 x 10⁹ cfu/ml, about 2.5 x 10⁹ cfu/ml, or about 5 x 10⁹ cfu/ml, about 1 x 10¹⁰ cfu/ml, about 1.5 x 10¹⁰ cfu/ml, about 2.5 x 10¹⁰ cfu/ml, about 5 x 10¹⁰ cfu/ml or about 1 x 10¹¹ cfu/ml.

[00067] Also contemplated by the present disclosure is the use of variants of the Lactobacillus species described herein. As used herein, the term "variant" refers to both naturally occurring and specifically developed variants or mutants of the species disclosed and exemplified herein. Variants may or may not have the same identifying biological characteristics of the specific species exemplified herein, provided they share similar advantageous properties in terms of treating or preventing poor sleep quality. Illustrative examples of suitable methods for preparing variants exemplified herein include, but are not limited to, gene integration techniques such as those mediated by insertional elements or transposons or by homologous recombination, other recombinant DNA techniques for modifying, inserting, deleting, activating or silencing genes, intraspecific protoplast fusion, mutagenesis by irradiation with ultraviolet light or X-rays, or by treatment with a chemical mutagen such as nitrosoguanidine, methylmethane sulfonate, nitrogen mustard and the like, and bacteriophage-mediated transduction. Suitable and applicable methods are well known in the art and are described, for example, in J. H. Miller, Experiments in Molecular Genetics, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1972); J. H. Miller, A Short Course in Bacterial Genetics, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1992); and J. Sambrook, D. Russell, Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2001), inter alia.

[00068] Also encompassed by the term “variant” as used herein are microbial strains phylogenetically closely related to species disclosed herein and strains possessing substantial sequence identity with the species disclosed herein at one or more phylogenetically informative markers such as rRNA genes, elongation and initiation factor genes, RNA polymerase subunit genes, DNA gyrase genes, heat shock protein genes and rec A genes. For example, the 16S rRNA genes of a “variant” strain as contemplated herein may share about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with a strain disclosed herein.

[00069] The Lactobacillus species described herein, and combinations thereof, or culture supernatants or cell free filtrates derived from culture media are typically administered in accordance with the present disclosure in the form of a composition. In embodiments in which combinations of species, or culture supernatants or cell free filtrates derived from culturing multiple species, those skilled in the art will appreciate that each of the species, supernatants or filtrates to be administered need not be contained in the same composition. Where administration is separate, administration may be sequential or simultaneous.

[00070] Compositions for use in accordance with the present disclosure may be prepared by admixing the relevant components and formulating the resulting mixture into a dosage form that is suitable for administration to a subject. Accordingly, the compositions may comprise pharmaceutically acceptable carriers, diluents, excipients and/or adjuvants. The carriers, diluents, excipients and adjuvants must be "acceptable" in terms of being compatible with other components of the composition, and not deleterious to the subject who is to receive the composition.

[00071] Compositions for administration in accordance with the present disclosure may be administered in any suitable form, typically by oral administration. The composition may be in unit dosage form. The term "unit dose" refers to a physically discrete unit of a dosage form suitable for use in humans, each unit containing a predetermined quantity of the composition to provide an effective amount. The compositions may be administered in any suitable form, typically in solid or liquid form. For example, the compositions may be formulated using methods and techniques well known to those skilled in the art, into tablets, troches, capsules, caplets, elixirs, suspensions, syrups, wafers, granules, powders, gels, pastes, solutions, creams, sprays, suspensions, soluble sachets, lozenges, effervescent tablets, chewable tablets, multi-layer tablets, and the like.

[00072] By way of example the composition may be provided to the user in a powder form, suitable for mixing by the user into any type of drink or food product (for example water, fruit juice or yoghurt) or for consumption as a powder in the absence of a drink or additional food product. The composition may be conveniently incorporated in a variety of food and/or beverage products, nutraceutical products, supplements, food additives, and over-the-counter formulations. The food or food additive may be a solid form such as a powder, or a liquid form. Specific examples of the types of beverages or foods include, but are not limited to water-based, milkbased, yoghurt-based, other dairy-based, milk-substitute based such as soy milk or oat milk, or juice-based beverages, water, soft drinks, carbonated drinks, and nutritional beverages, (including a concentrated stock solution of a beverage and a dry powder for preparation of such a beverage); baked products such as crackers, breads, muffins, rolls, bagels, biscuits, cereals, bars such as muesli bars, health food bars and the like, dressings, sauces, custards, yoghurts, puddings, pre-packaged frozen meals, soups and confectioneries.

[00073] In particular embodiments, compositions to be administered in accordance with the present disclosure may be used as nutritional supplements to help subjects who suffer from mild, moderate or severe sleep disturbance improve the quality of their sleep.

[00074] As will be appreciated by those skilled in the art, the choice of pharmaceutically acceptable carrier or diluent may be readily determined by a person skilled in the art. A person skilled in the art will readily be able to determine appropriate formulations useful in the methods of the disclosure using conventional approaches.

[00075] Solid forms for oral administration may contain binders acceptable in pharmaceutical practice, sweeteners, disintegrating agents, diluents, flavourings, coating agents, preservatives, lubricants and/or time delay agents. Suitable binders include gum acacia, gelatine, corn starch, gum tragacanth, sodium alginate, carboxymethylcellulose or polyethylene glycol. Suitable sweeteners include sucrose, lactose, glucose, stevia, xylitol, aspartame or saccharine. Suitable disintegrating agents include corn starch, methylcellulose, polyvinylpyrrolidone, guar gum, xanthan gum, bentonite, alginic acid or agar. Suitable diluents include lactose, sorbitol, mannitol, dextrose, kaolin, cellulose, calcium carbonate, calcium silicate or dicalcium phosphate. Suitable flavouring agents include peppermint oil, oil of Wintergreen, cherry, orange or raspberry flavouring. Suitable coating agents include polymers or copolymers of acrylic acid and/or methacrylic acid and/or their esters, waxes, fatty alcohols, zein, shellac or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable time delay agents include glyceryl monostearate or glyceryl distearate.

[00076] Liquid forms for oral administration may contain a liquid carrier. Suitable liquid carriers include water, oils such as olive oil, peanut oil, sesame oil, sunflower oil, safflower oil, arachis oil, coconut oil, liquid paraffin, ethylene glycol, propylene glycol, polyethylene glycol, ethanol, propanol, isopropanol, glycerol, fatty alcohols, triglycerides or mixtures thereof. Suspensions for oral administration may further comprise dispersing agents and/or suspending agents. Suitable suspending agents include sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose, poly-vinyl-pyrrolidone, sodium alginate or acetyl alcohol. Suitable dispersing agents include lecithin, polyoxyethylene esters of fatty acids such as stearic acid, polyoxyethylene sorbitol mono- or di-oleate, -stearate or -laurate, polyoxyethylene sorbitan mono- or di-oleate, -stearate or -laurate and the like, emulsions for oral administration may further comprise one or more emulsifying agents. Suitable emulsifying agents include dispersing agents as exemplified above or natural gums such as guar gum, gum acacia or gum tragacanth.

[00077] In exemplary embodiments, each dose of a liquid composition may comprise about 1 ml to about 25 ml liquid formulation of Lactobacillus at a final concentration of between about 10⁵ and 10¹¹ cfu/ml. The volume of the liquid formulation may be, for example, about 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8ml, 9 ml, 10 ml, 11 ml, 12 ml, 13 ml, 14 ml, 15 ml, 16 ml, 17 ml, 18 ml, 19 ml, 20 ml, 21 ml, 22 ml, 23 ml, 24 ml, or 25 ml.

[00078] Compositions for use in accordance with the present disclosure may comprise one or more prebiotic components. Suitable prebiotics include, for example, polydextrose, inulin, fructooligosaccharides (FOS), xylooligosaccharides (XOS), galactooligosaccharides (GOS), mannan oligosaccharides, protein-based green lipped mussel extract, and various prebiotic - containing foods such as raw onion, raw leek, raw chickory root and raw artichoke. In certain embodiments the prebiotic is a fructooligosaccharide.

[00079] In certain embodiments, the methods of the present disclosure may be employed as part of a broader regime to promote and maintain a state of sleep in an individual. As such, the present methods may be used in conjunction with other methods known to promote and maintain a state of sleep. Additionally, compositions for use in accordance with the present disclosure may incorporate additional ingredients known to promote and maintain a state of sleep.

[00080] The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

[00081] The present disclosure will now be described with reference to the following specific examples, which should not be construed as in any way limiting the scope of the invention.

Examples

[00082] The following examples are illustrative of the invention and should not be construed as limiting in any way the general nature of the disclosure of the description throughout this specification.

Example 1

[00083] A 46 year old male had a long history of disrupted sleep starting about the time of the birth of his first child, 10 years ago. He was able to fall asleep but would wake easily during the night, most often around 3am and then very rarely return to sleep, or if at all, only after 5-5:30am. This sleeping pattern occurred usually 5 out of 7 nights a week in a normal week and worse during times of stress.

[00084] The subject consumed 10ml of Lactobacillus plantarum SVT 09P1 live bacteria (formulated in a composition comprising water, sodium chloride, xylitol and raspberry flavour) at a concentration of 1 x 10⁹ CFU/ml twice a day, once in the morning and once in the evening, for six weeks. After 10 days a noticeable change occurred in sleeping pattern, including both an undisturbed sleep (i.e. no more waking in the night) and the sleep quality itself improved, feeling deep and restful. This improvement in sleep quality continued throughout the six week period of consumption of the L. plantarum. Subsequently two weeks after ceasing to take the bacteria, the subject’s sleep quality commenced to deteriorate, and occasionally insomnia would occur in a similar manner to previously, namely waking during the night around 3am and struggling to return to sleep. Six weeks after stopping taking the bacteria, sleep pattern continues to deteriorate, though is still notably better than prior to taking the bacteria.

Example 2

[00085] A 38 year old female had a 15 year history of chronic insomnia that had started with a stressful event. Over this time she had tried numerous supplements and medications none of which provided effective management of her insomnia, some having only short term effects or requiring increasing doses to reach any form of efficacy. The patterns of insomnia varied over the 15 years, ranging from not being able to go to sleep at all or only at around 4am, to being able to fall asleep only to wake around 12am - lam and not be able to go back to sleep. Stress made the insomnia worse.

[00086] The subject consumed 10 mL of a combination of Lactobacillus zeae SVT 08Z1, Lactobacillus paracasei SVT 04P1 and Lactobacillus parafarraginis SVT 05P2 (formulated in saline water) at a combined concentration of 1.5 x 10^uCFU/mL twice a day (morning and evening) for 14 days. She noticed a significant improvement in her sleep after only about 3 days of consuming the product such that she was able to cease all other sleeping medications and was able to enjoy a long and restful sleep each night, averaging about 8 hours per night. She was able to fall asleep quickly and when she woke during the night she was able to easily go straight back to sleep. This had not been possible over the previous 15 years. The effect of the product on her sleeping patterns was still noticeable many weeks after ceasing consumption, with her sleeping pattern starting to resume back to mild insomnia after approximately four weeks, mainly waking during the night and not being able to get back to sleep.

Example 3

[00087] 42 male and female subjects, aged between 32 and 81 years, and with self-reported poor sleep quality were included in a study investigating improvements in sleep quality with Lactobacillus zeae, Lactobacillus plantarum, or a combination of both L. zeae and L. plantarum. Subjects were divided into three groups (randomised based on their initial score using the Pittsburgh Sleep Quality Index):

• Group A (n=14): L. zeae SVT 08Z1, administered at 1 x 10¹⁰ CFU per 10 ml dose, twice daily;

• Group B (n=14): L. zeae SVT 08Z1 and L. plantarum SVT 09P1, administered at 1 x 10¹⁰ CFU per 10 ml dose, twice daily;

• Group C (n=14): L. plantarum SVT 09P1, administered at 1 x 10¹⁰ CFU per 10 ml dose, twice daily.

[00088] The validated Pittsburgh Sleep Quality Index (PSQI, modified to refer to weekly observations), containing 19 self-rated questions, was used to assess sleep quality on a weekly basis for five weeks. The questions were combined to form seven “component” scores, each of which has a range of 0-3 points. In all cases, a score of “0” indicates no difficulty, while a score of “3” indicates severe difficulty. The seven component scores were then added to yield one “global” score, with a range of 0-21 points, “0” indicating no difficulty and “21” indicating severe difficulties in all areas. Additional questions were included to capture general wellbeing, comments on what may be impacting a subject’s sleep and comments about their experience with the product administered. The comment on their experience was only included at time point 3. The questionnaire was sent to subjects using SurveyMonkey.

[00089] Figures 1 to 4 provide weekly mean, or change in mean, PSQI scores and general wellbeing scores for each treatment group across the five weeks of treatment. Significance values (p-values) are also shown for each treatment group compared to baseline (scores prior to commencement of treatment). All three treatments resulted in improved sleep quality, as assessed by reduced mean PSQI scores (Figures land 2), and improved general wellbeing (Figures 3 and 4) each week over the five weeks of treatment. Over the weekly analysis, the treatment in Group A appeared to be more effective, however when analyszed across the five weeks (Figure 5), all treatment groups demonstrated a significant improvement in sleep quality.

[00090] Participants from each group were encouraged to comment on their reaction and feelings before and/or after sleep. The comments below are from week 3.

Group A

Male, 36: swelling i

Female, 48: Yes, I can notice I am going to sleep much easier than before. I also noticed the quality of my sleep is deeper and even though I still wake up in the early morning, I feel the hours I was asleep were a deeper sleep feeling, not a light sleep like before. I sometimes feel it is very difficult to get out of bed as I feel the need to keep sleeping for longer.

Male, 65: I have been finding it slightly easier to get back to sleep in the middle of the night.

Female, 74: I feel I am having more uninterrupted sleep during the last week.

Female, 41: but have a lo

Female, 33: Recently feeling less bloated in mornings. I am feeling more awake and clearer upon waking.

Female, 49: I am now seeing a pattern emerging of better-quality sleep. Last night I slept from ll:45ish through to 6am. This has not happened to me in years... even decades!

Male, 63: Something is definitely going on. Sleep seems more profound although I’ve had more of a feeling, I need further catch-up rather than a feeling I haven’t slept well.

Female, 45: When I take it consistently, I am so drowsy when bedtime comes round, whereas before I started the program, I would still be completely wide awake not at all sleepy. I love it!

Female, 43: Stay asleep without tossing and turning.

Male, 48: Seems to slowly help out. Group B

Male, 58: Feeling more relaxed.

Female, 53: Sleeping heavier.

Female, 45: Initially I felt there was a definite improvement, both in the time it took to get to sleep and the soundness of my sleep. However, this week was not so great unfortunately.

Female, 41 : I think I am getting a deeper sleep and falling asleep much faster.

Male, 52: Still waking up but period before returning to sleep has reduced. Aware of dreams. Period to initially get off to sleep has reduced.

Male, 53: Weight loss seems to be feeling better.

Group C

Male, 72: I often wake up during the night and have trouble getting back to sleep. I am now finding that I go back to sleep quite quickly, and I am definitely getting extra sleep.

Female, 56: Probably more deep sleep - intuitively I feel this.

Male, 53: the mornin

Female, 61: I feel more relaxed in general.

Female, 39: I feel the quality of my sleep has improved. It takes me a long time to fall asleep, but I feel it is a little more quickly.

Male, 53: Sleep is slightly improving.

Female, the day

Female, 69: Last week has been the best sleep I have had in a long while.

Example 4

[00091] A phase I/II randomised, double-blind, placebo-controlled study is undertaken to assess safety and efficacy of a live biotherapeutic product comprising L. z.eae and L. plantarum, in subjects with clinically diagnosed insomnia. The biotherapeutic product administered in a 20 mL dose comprising 5.0 x 10¹⁰ CFU/dose of L. z.eae and L. plantarum (2.5 x 10¹⁰ CFU/dose of each of L. zeae SVT 08Z1 and L. plantarum SVT 09P1) suspended in 0.9% saline (with added flavour and stevia).

[00092] Inclusion criteria for the study include: male and female participants aged 18-70 years; an Insomnia Severity Index (ISI) score greater than or equal to 8 with insomnia symptoms for more than 3 times per week and present for longer than 3 months; and a clinical diagnosis of insomnia determined by clinical interview conducted by a trained sleep psychologist. Exclusion criteria include: sleep apnoea (AHI > 10) determined by polysomnography; a score of > 4 on the STOP-bang questionnaire; a medical condition, medication of sleep-related movement disorder that may be causative of insomnia symptoms; shift worker working hours between 10pm and 4am; history of major psychiatric disorder in the past 12 months.

[00093] 50 participants are selected and, following a 14 day screening period, are randomised into two groups of 25, Group A receiving a placebo, and Group B receiving the biotherapeutic product. Treatment is over 35 days during which the biotherapeutic product (or placebo) is administered orally twice a day. Safety and tolerability are assessed by incidence of adverse events, serious adverse events and treatment emergent adverse events during the treatment period.

[00094] Endpoints assessed include:

• Improvement of subjective symptoms of insomnia as assessed by the Insomnia Severity Index (ISI) during the treatment period;

• Improvement of Sleep Quality as determined objectively using the Somfit measurement outputs: Sleep Onset Latency (SOL), Wake After Sleep Onset (WASO), Total Sleep Time (TST), Sleep Efficiency (SE) and number of awakenings (nWAK) and number of awakenings per hour of sleep (WAKI), comparing the average measures for each week (up to the end of the treatment period, Day 35) to the baseline average measure (taken during screening period);

• Change in sleep architecture, determined by time spent in sleep states, REM and Nl, N2 and N3 using the Somfit, comparing the average measure for each week (up to the end of the treatment period, Day 35) to the baseline average measure (taken during screening period);

• Change in rate of arousal events, both total and hourly, ascertained using the Somfit, comparing the average measure for each week (up to the end of the treatment period, Day 35) to the baseline average measure (taken during screening period);

• Improvement of sleep quality as determined subjectively from patient reporting using the sleep diary with the following metrics: SOL, WASO, TST, SE, nWAK and participant reported sleep satisfaction, comparing the average measure for each week (up to the end of the treatment period, Day 35) to the baseline average measure (reported during screening period);

• Improvement in sleep quality determined by the Pittsburgh Sleep Quality Index (PSQI) questionnaire comparing Day 35 to baseline;

• Improvement in daytime functioning assessed by the Work and Social Adjustment Scale (WSAS) at Day 35 compared to baseline;

• Improvement in quality of life assessed by the Item 36 Short Form Survey (SF-36) at Day 35 compared to baseline;

• Improvement in daytime sleepiness assessed by the Epworth Sleepiness Scale (ESS) at Day 35 compared to baseline;

• Change in DASS21 outcome measures from Baseline to Day 35;

• Comparison of efficacy between participants with threshold insomnia score compared to participants with subthreshold insomnia (determined by ISI) for all objective (Somfit) and Subjective (ISI, PSQI, sleep diary) measures;

• Comparison in change in quality of life and daytime functioning as assessed by the SF- 36, ESS and the WSAS questionnaires respectively, between participants with threshold insomnia score compared to participants with subthreshold insomnia (based on ISI);

• Comparison of overall changes in the composition of the participants gastrointestinal microbiome induced by SVT-4A1011 at Day 35 compared to baseline values.

Deposit Details

[00095] Details of the biological material deposited pursuant to the Budapest Treaty are provided hereinbefore in the specification.

[00096] Lactobacillus plantarum SVT 09P1 was deposited pursuant to the Budapest

Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM), Federal Public Planning Service Science Policy, 8 rue de la Science B-1000, Brussels, Belgium, on 10 August 2020 under Accession Number LMG P-31923.

[00097] The following Lactobacilli have been described previously, for example in

WO 2020/073088, the disclosure of which is incorporated herein in its entirety. In summary:

• Lactobacillus zeae SVT 08Z1 was deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM), Federal Public Planning Service Science Policy, 8 rue de la Science B-1000, Brussels, Belgium, on 27 February 2019 under Accession Number LMG P-31295.

• Lactobacillus parafarraginis SVT 05P2 was deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM), Federal Public Planning Service Science Policy, 8 rue de la Science B-1000, Brussels, Belgium, on 27 February 2019 under Accession Number LMG P-31292.

• Lactobacillus buchneri SVT 06B1 was deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM), Federal Public Planning Service Science Policy, 8 rue de la Science B-1000, Brussels, Belgium, on 27 February 2019 under Accession Number LMG P-31293.

• Lactobacillus rapi SVT 07R1 was deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM), Federal Public Planning Service Science Policy, 8 rue de la Science B-1000, Brussels, Belgium, on 27 February 2019 under Accession Number LMG P-31294.

• Lactobacillus paracasei SVT 04P1 was deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Micro-organisms (BCCM), Federal Public Planning Service Science Policy, 8 rue de la Science B-1000, Brussels, Belgium, on 27 February 2019 under Accession Number LMG P-31290.

• Lactobacillus diolivorans SVT 01D1 was deposited pursuant to the Budapest Treaty with the Belgian Coordinated Collections of Microorganisms (BCCM), Federal Public Planning Service Science Policy, 8 rue de la Science B-1000, Brussels, Belgium, on 27 February 2019 under Accession Number LMG P-31287.

Claims

23 Claims

1. A method for inducing or maintaining sleep in a human subject, comprising administering to the subject an effective amount of Lactobacillus plantarum and/or Lactobacillus zeae, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured.

2. A method for improving sleep quality in a human subject, comprising administering to the subject an effective amount of Lactobacillus plantarum and/ or Lactobacillus zeae, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured.

3. A method for treating insomnia in a human subject, comprising administering to the subject an effective amount of Lactobacillus plantarum and/or Lactobacillus zeae, or a culture supernatant(s) or cell free filtrate(s) derived from culture media in which said Lactobacillus species has been cultured.

4. A method according to claim 2, wherein improving sleep quality comprises improving one or more of the onset of sleep, sleep depth, sleep duration, or fatigue and/or lethargy following sleep.

5. A method according to claim 3, wherein the insomnia is sleep onset insomnia, sleep maintenance insomnia, early morning awakening, or transient insomnia.

6. A method according to any one of claims 1 to 5, wherein the method comprises administering a composition comprising live Lactobacillus plantarum.

7. A method according to any one of claims 1 to 5, wherein the method comprises administering a composition comprising live Lactobacillus zeae.

8. A method according to any one of claims 1 to 5, wherein the method comprises administering a composition comprising live Lactobacillus plantarum and Lactobacillus zeae.

9. A method according to any one of claims 1 to 8, wherein the method further comprises administering one or more additional agents.

10. A method according to claim 9, wherein the one or more additional agents comprise one or more additional Lactobacillus species or a culture supernatant or cell free filtrate derived from culture media in which said Lactobacillus species has been cultured.

11. A method according to claim 10, wherein the one or more additional Lactobacillus species is selected from Lactobacillus buchneri, Lactobacillus rapi, Lactobacillus paracasei, Lactobacillus parafarraginis, and Lactobacillus diolivorans.

12. A method according to any one of claims 1 to 5, wherein the method comprises administering a composition comprising live Lactobacillus zeae, Lactobacillus paracasei and Lactobacillus parafarraginis.

13. A composition comprising Lactobacillus plantarum and/or Lactobacillus zeae, or a culture supernatant(s) or cell free filtrates(s) derived from culture media in which said Lactobacillus species has been cultured, when used for inducing or maintaining sleep or for improving sleep quality.

14. A composition according to claim 13, comprising live Lactobacillus plantarum and Lactobacillus zeae.