An eurycoma longifolia extract for use in the treatment of insomnia or for improving mood, quality of life, or working memory

Abstract

Insomnia is a widespread health issue affecting cognitive abilities. Eurycoma Longifilia (EL), a Southeast Asian adaptogen herb known for its aphrodisiac use. This open-label clinical human trial examined EL's impact on treating insomnia looking at sleep, cognitive functioning, and psychological well-being. 46 healthy participants aged 21-41, with Insomnia Severity Index (ISI) score ≥8 took one 200mg EL extract capsule daily for 6 weeks and completed ≥ 90% of dosage. Assessments including ISI, Inquisit Stroop task, Digitspan, WHO-QOL were done at pre-intervention (T0), 3 weeks (T1), 6 weeks (T2), and 10 weeks (T3). Differences for assessments between each timepoints and T0 were assessed using Linear Mixed Model analysis with participants as a random effect. EL improved psychological well-being, sleep quality and consequently cognitive flexibility, working memory.

Description

DESCRIPTIONTITLE OF INVENTION: AN EURYCOMA LONGIFOLIA EXTRACT FOR USE IN THE TREATMENT OF INSOMNIA OR FOR IMPROVING MOOD, QUALITY OF LIFE, OR WORKING MEMORYREFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority to Singapore patent application No. 10202402298Q, filed 1 August 2024, the contents of which are incorporated herein by reference.TECHNICAL FIELD

[0002] The present disclosure relates generally to agents for use in the treatment of insomnia or improving the mood, quality of life, or working memory of an individual in need.BACKGROUND

[0003] The following discussion of the background to the invention is intended to facilitate an understanding of the present invention only. It should be appreciated that the discussion is not an acknowledgement or admission that any of the material referred to was published, known or part of the common general knowledge of the person skilled in the art in any jurisdiction as at the priority date of the invention.

[0004] Sleep is essential for animals to function in everyday life. Having insufficient sleep for just one night can impact our cognitive abilities and mood the next day. Chronic sleep deprivation clinically referred to as insomnia is a sleep disorder. Insomnia can increase the risk of developing physical and mental health conditions such as heart disease, strokes, diabetes and depression (Hanson & Huecker, 2019 StatPearls Publishing; Treasure Island). However, sleep difficulties have become increasingly common due to a variety of reasons including poor sleep hygiene and stress (Kronholm et al, 2008 J sleep Res. 17 (1):54-62;). Insomnia may have different manifestations including difficulty falling asleep, or difficulty staying asleep for as long as desired. A common treatment is prescription of sleeping pills or psychoactive drugs such as benzodiazepines, non-benzodiazepines and barbiturates. However, such treatments are sometimes associated with injuries, dementia and addiction. Insomnia is a widespread health issue affecting cognitive abilities.

[0005] The root of the Eurycoma longifolia plant has traditionally been known in Southeast Asian medicine as Tongkat All and is traditionally used for libido related difficulties. Today, this root extract is usually marketed as an all-natural alternative to Hormone Replacement Therapy to address symptoms of aging that are associated with low testosterone levels. Tongkat Ali has previously been used to improve sexual function a Southeast Asian adaptogen herb known for its aphrodisiac use. Previous studies investigating its use have found a consistent effect on testosterone levels and sexual function, these previous studies all targeted older populations with infertility, hypogonadism or erectile dysfunction symptoms, and there is evidence that Tongkat Ali increases libido in those with subnormal testosterone levels, while the libido of healthy individuals would simply be maintained (Chen et al, 2014 Phytother Res. 28(4):544-50).

[0006] There exists a need to alleviate at least one of the aforementioned problems.SUMMARY

[0007] Alternative agents for use in the treatment of insomnia are envisaged.

[0008] Accordingly, an aspect of the invention refers to a method of treating an individual diagnosed with insomnia or improving the mood, quality of life, or working memory of an individual in need comprising administering to the individual diagnosed with insomnia or providing to the individual in need a root extract of a Eurycoma longifolia plant.

[0009] According to another aspect of the invention there is a root extract of a Eurycoma longifolia plant for use in the treatment of insomnia.

[0010] According to another aspect of the invention there is use of a root extract of a Eurycoma longifolia plant in the manufacture of a medicament for use in the treatment of insomnia.

[0011] Other aspects and features of the present invention will become apparent to those of ordinary skill in the art upon review of the following description of specific embodiments of the invention in conjunction with the accompanying figures.BRIEF DESCRIPTION OF THE DRAWINGS

[0012] In the figures, which illustrate, byway of non-limiting examples only, embodiments of the present invention,

[0013] [Fig. 1]: A) A schematic of the Test protocol for the treatment regimen. B) gender distribution of the participants in the trial.

[0014] [Fig. 2]: (A) summary of the subjective measurements taken (i) ISI, (ii) ESS, (iii) POMS-2 and (iv) WHO-QOL (B) treatment significantly improved sleep and reduced insomnia and chances of daytime sleepiness measured by (i) ISI and (ii) ESS (C-D) treatment improved physical and psychological quality of life, probably mediated by better sleep measured by (iii) POMS-2 or (E-H) measured by (iv) WHO-QOL.

[0015] [Fig. 3]: (A) Summary of the cognitive measurements taken (v) STOOP and (iv) DIGI-SPAN. (B & C) Treatment improved cognitive performance measured by (v) STOOP and (D) (iv) DIGI-SPAN.DETAILED DESCRIPTION

[0016] Throughout this document, unless otherwise indicated to the contrary, the terms “comprising”, “consisting of’, “having" and the like, are to be construed as non-exhaustive, or in other words, as meaning “including, but not limited to”.

[0017] Furthermore, throughout the document, unless the context requires otherwise, the word “include” or variations such as “includes” or “including” will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0018] Throughout the description, it is to be appreciated that the term ‘treat’, ‘treatment’ and its plural form include reducing sleeping difficulties and / or promoting sleep quality and its associated outcomes of cognition, mood, and overall quality of life.

[0019] Unless defined otherwise, all other technical and scientific terms used herein have the same meaning as is commonly understood by a skilled person to which the subject matter herein belongs.

[0020] According to various embodiments, there is a method of treating an individual diagnosed with insomnia or improving the mood, quality of life, or working memory of an individual in need comprising administering to the individual diagnosed with insomnia or providing to the individual in need a root extract of a Eurycoma longifolia plant.

[0021] As used herein, an individual diagnosed with insomnia may be any human experiencing chronic sleep disorder or having less than 7 hours of sleep for one or more nights per week over an extended period of 2-3 months. In various embodiments insomnia may be as a result of the individual having a hard time falling asleep. In various embodiments insomnia may be as a result of the individual waking up during the night, sometimes for an extended period. In various embodiments insomnia may be as a result of the individual waking up too early. In various embodiments insomnia may be diagnosed by a medical professional.

[0022] As used herein, an individual in need may be an individual in need of an improved mood, an individual in need of improved quality of life or an individual in need of an improvement in workingmemory. In various embodiments an individual in need of improved mood, improved quality of life, or improved working memory may be identified by a POMS, and / or WHO-QOL survey.

[0023] In various embodiments, the mood of an individual in need of improved mood may be determined by changes in the individuals mood and emotional states measured using a 35-item Profile of Mood States 2 (POMS-2) questionnaire. In various embodiments, the mood of an individual in need of improved mood may be determined by measuring any one of the feelings of anger, sadness or hopelessness and the improvement may similarly be determined by measuring reduced anger or reduced sadness or hopelessness after taking the root extract of a Eurycoma longifolia plant. In various embodiments, the mood of an individual in need of improved mood may be characterized by any one of 7 different mood states comprising Anger-Hostility, Tension-Anxiety, Confusion-Bewilderment, Vigour- Activity, Fatigue-Inertia, Depression-Dejection, and / or Friendliness and the improvement may similarly be characterized by a change in any one of 7 different mood states in the individual in need after taking the root extract of a Eurycoma longifolia plant. In various embodiments, the mood of an individual in need of improved mood may be determined by Mood Disturbance scores calculated from the mood states (excluding friendliness) and the improvement may similarly be characterized by a change in the Mood Disturbance scores indicating an improvement in the mood of the individual in need. In various embodiments, the mood of an individual in need of improved mood may be determined by Total Mood Disturbance scores calculated from the mood states (excluding Friendliness) and the improvement may similarly be characterized by a change in the Total Mood Disturbance scores indicating an improvement in the mood of the individual in need. In various embodiments, the mood of an individual in need of improved mood may be determined by deep sleep measurements and the improvement may similarly be characterized by in increase in deep sleep measurements.

[0024] In various embodiments, the quality of life of an individual in need of improved quality of life may be measured using the World Health Organisation Quality of Life (WHO-QOL) questionnaire. In various embodiments, the quality of life of an individual in need of improved quality of life may bedetermined by measuring any one of Pain and Discomfort, Positive Thinking, negative thinking Social Support or the other measurements determined in the WHO-QOL and the improvement may similarly be determined by measuring changes in the WHO-QOL questionnaire by the individual in need after taking the root extract of a Eurycoma longifolia plant.

[0025] In various embodiments, the working-memory of an individual in need of improved workingmemory may be measured using the World Health Organisation Quality of Life (WHO-QOL) questionnaire. In various embodiments, the working-memory of an individual in need of improved working-memory may be determined by measuring any one of Thinking, Learning, Memory and Concentration measurements determined in the WHO-QOL and the improvement may similarly be determined by measuring changes in the WHO-QOL questionnaire by the individual in need after taking the root extract of a Eurycoma longifolia plant. In various embodiments, the working-memory of an individual in need of improved working-memory may be determined by a cognitive performance task such as a Digit Span (Forward and Backward recall) assessment whereby the performance of the individual in need of improved working-memory increases after taking the root extract of a Eurycoma longifolia plant. In various embodiments, the working-memory of an individual in need of improved working-memory may be determined by a Stroop Task assessment whereby the performance of the individual in need of improved working-memory demonstrates a reduced overall mean response time after taking the root extract of a Eurycoma longifolia plant. A reduced overall mean response time after taking the root extract of a Eurycoma longifolia plant was observed in all individuals across all timepoints regardless of gender and working status. This indicated an objective improvement in cognitive flexibility after taking the root extract of a Eurycoma longifolia plant.

[0026] The root extract of a Eurycoma longifolia plant was found to reduce insomnia and improve sleep in individual diagnosed with insomnia administered with the root extract. In various embodiments, the root extract of a Eurycoma longifolia plant was found to reduce insomnia and improve sleep in individual diagnosed with insomnia administered with the root extract for 3 weeks. In various embodiments, the root extract of a Eurycoma longifolia plant was found to reduce insomnia and improve sleep in individual diagnosed with insomnia administered with the root extract for 6 weeks. In various embodiments, the root extract of Eurycoma longifolia plant was found to reduce insomnia and improve sleep in individual diagnosed with insomnia for at least 4 weeks after being administered the root extract for 6 weeks. The root extract of a Eurycoma longifolia plant was found to improve sleep indicators across different sex and working status.

[0027] In various embodiments the root extract of a Eurycoma longifolia plant may be administered orally, nasally, enterally, or parenterally.

[0028] In various embodiments, the root extract of a Eurycoma longifolia plant comprises about 1 .5% Eurycomanone, about 22% protein, about 30% polysaccharides and about 35% glycosaponin. As the extracts are from plants, it may be understood that it may be difficult to standardize the bioactive content and quality of the extracts. These standardization markers: eurycomanone; total protein; total polysaccharide; and glycosaponin are used for E. longifolia root extracts to ensure standard quality of the extracts. In various embodiments to further ensure quality the plants are cultivated in preferential growing conditions of high humidity, 2 hours of high light separating 5 hours each of medium light before and afterthe highlight, in well drained sandy soils. In various embodiments, to further ensure quality the roots are grown in bioreactor to enhance the production of the bioactives. In various embodiments, the root extract of a Eurycoma longifolia plant may comprise saponins, alkaloids, polyphenols, coumarin, tannins, triterpenes and the glycoproteins eurycomanol, eurycomanone and eurycomalactone.

[0029] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated for oral administration. This may include other forms of enteral administration including, buccal, sulabial, and sublingual. In various embodiments, the root extract of a Eurycoma longifolia plant is formulated for oral administration may be in the form of tables, capsules, powders, granuels, teas, drops, liquid medication, or any other form that will allow enteral administration.

[0030] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated for oral consumption. In various embodiments, this may include in the form of a supplement, a food preparation such as a functional food, a beverage such as a drink product or any other formulation suitable for oral consumption.

[0031] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a capsule for oral administration. In various embodiments, the capsule is formed of a coating that dissolves in the stomach. In various embodiments, the coating may be formed of a gelling agent such as gelatin or cellulose, plasticizers such as glycerin or sorbitol, and other additives such as colouring agents, preservatives, disintegrants, or lubricants. In various embodiments, the capsule is formed of a hard- shelled capsule preferably made by extrusion. In various embodiments, the capsule is formed of a soft- shelled capsule

[0032] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a liquid for oral administration. In various embodiments, the liquid is formed as a decoction, emulsion, herbal tea, hydrogel, syrup, spray, or inhalant.

[0033] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a supplement for oral consumption. In various embodiments, the term “supplement" refers generally to a product that contains substances like vitamins, minerals, nutrients, botanicals, amino acids that are intended to supplement food and / or drink consumed or ingested by an individual such as an individual in need to sustain growth, repair vital processes or provide energy. In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a supplement for oral consumption with other products that may contain substances like vitamins, minerals, nutrients, botanicals, and / or amino acids. In various embodiments, the root extract of a Eurycoma longifolia plant comprises less than 200mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 150mg and 200mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 100mg and 200mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 200mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 200mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 200mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 200mgin a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 200mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 10Omg and 150mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 150mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 150mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 150mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 150mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 150mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 100mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 100mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 100mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 100mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 100mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 75mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 75mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 75mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 75mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 50mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 50mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 50mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 25mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 25mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 20mg in a supplement formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant in a supplement is formulated for multiple dosages such as 2, 3, 4, 5, 6, 7, 8, 9, 10, or 15 dosages daily.

[0034] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as food preparation for oral consumption. In various embodiments, the term “food preparation” refers generally to material of either plant or animal origin, or of synthetic sources, that contain elements such as essential nutrients, carbohydrates, protein, fat, vitamin, mineral that may be consumed or ingested by an organism such as an individual to sustain growth, repair vital processes or provide energy. In various embodiments,the root extract of a Eurycoma longifolia plant is formulated as food preparation for oral consumption with other material of either plant or animal origin, or of synthetic sources, that may contain elements such as essential nutrients, carbohydrates, protein, fat, vitamin, and / or mineral. In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as food preparation for oral consumption with chocolate. In various embodiments, the root extract of a Eurycoma longifolia plant comprises less than 200mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 150mg and 200mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 100mg and 200mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 200mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 200mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 200mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 200mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 200mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 100mg and 150mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 150mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 150mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 150mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 150mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 150mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 100mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 100mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 100mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 100mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 100mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 75mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 75mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 75mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 75mg in a food preparation formulated for oral consumption. In variousembodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 50mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 50mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 50mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 25mg in a food preparation formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant in a food product is formulated for multiple dosages such as 2, 3, 4, 5, 6, 7, 8, 9, 10, or 15 dosages daily.

[0035] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a beverage for oral consumption. In various embodiments, the term “beverage” refers generally to a liquid for drinking. In various embodiments the liquid may be water, flavoured water, soft drinks, alcoholic drinks, health drinks, an enriched drink, a diary-based drink such as milk or yogurt a fruit juice, a vegetable juice or any combination of these that may be consumed or ingested by an organism such as an individual in need. In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a beverage for oral consumption in any consumable liquid such as water, flavoured water, soft drinks, alcoholic drinks, health drinks, an enriched drink, a diary-based drink such as milk or yogurt a fruit juice, a vegetable juice or any combination of these. In various embodiments, the root extract of a Eurycoma longifolia plant less than comprises less than 200mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant less than comprises between 150mg and 200mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 100mg and 200mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 200mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 200mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 200mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 200mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 200mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 100mg and 150mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 150mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 150mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 150mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 150mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 150mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 75mg and 100mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 100mg in a beverage formulatedfor oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 100mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 100mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 100mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 50mg and 75mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 75mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 75mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 75mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 25mg and 50mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 50mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 50mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 20mg and 25mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 25mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant comprises between 15mg and 20mg in a beverage formulated for oral consumption. In various embodiments, the root extract of a Eurycoma longifolia plant in a beverage is formulated for multiple dosages such as 2, 3, 4, 5, 6, 7, 8, 9, 10, or 15 dosages daily.

[0036] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a single dose comprising 200mg of the Eurycoma longifolia root extract.

[0037] In various embodiments, 200mg of the root extract of a Eurycoma longifolia plant is administered daily.

[0038] According to various embodiments, there is a root extract of a Eurycoma longifolia plant for use in the treatment of insomnia.

[0039] A root extract of a Eurycoma longifolia plant has the advantage of being useful in the treatment of insomnia. Successfully improving sleep indicators, increase in total minutes of REM sleep, and improve 30-day-average of light, deep and REM sleep.

[0040] In various embodiments, the root extract for use in treating insomnia comprises about 1 .5% Eurycomanone, about 22% protein, about 30% polysaccharides and about 35% glycosaponin.

[0041] In various embodiments, the root extract for use in treating insomnia is formulated for oral administration.

[0042] In various embodiments, the root extract for use in treating insomnia is formulated for oral administration as a capsule or as a liquid.

[0043] In various embodiments, the root extract for use in treating insomnia is formulated as a dose comprising 200mg.

[0044] In various embodiments, the root extract for use in treating insomnia is formulated as a dailydose.

[0045] According to various embodiments, there is use of a root extract of a Eurycoma longifolia plant in the manufacture of a medicament for use in the treatment of insomnia.

[0046] The use of a root extract of a Eurycoma longifolia plant in the manufacture of a medicament has the advantage of being able to improve sleep indicators, increase in total minutes of REM sleep, improve 30-day-average of light, deep and REM sleep in the treatment of insomnia.

[0047] In various embodiments, the root extract comprises about 1 .5% Eurycomanone, about 22% protein, about 30% polysaccharides and about 35% glycosaponin.

[0048] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated to be administered orally.

[0049] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated to be administered orally as a capsule or as a liquid.

[0050] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a dose comprising 200mg.

[0051] In various embodiments, the root extract of a Eurycoma longifolia plant is formulated as a daily dose.

[0052] In various embodiments the root extract of a Eurycoma longifolia plant is comprised in a capsule. In various embodiments the root extract of a Eurycoma longifolia plant is formulated as a food product. In various embodiments the root extract of a Eurycoma longifolia plant is formulated as a beverage.

[0053] Examples

[0054] This study investigates the efficacy and potential side effects of using a root extract of a Eurycoma longifolia plant as a natural sleep treatment for insomnia. It may provide a safe supplement for individuals who experience trouble sleeping. This was examined through both subjective and objective measures of sleep quantity and quality, and associated outcomes including physiological (blood cortisol, serotonin, oxytocin) and psychological (cognition, mood) outcomes.

[0055] Dose

[0056] The root water extract of Eurycoma longifolia plant comprises about 1.5% Eurycomanone, about 22% protein, about 30% polysaccharides and about 35% glycosaponin. Participants will be taking one 200mg capsule (total 200mg of root extract from Eurycoma longifolia plant) daily for 6 weeks. Capsules of Eurycoma longifolia root extract (EL) are to be stored in a cool dry place below 30°C, avoiding direct sunlight and out of reach of children. The drug formulation is encased in a capsule and taken orally once a day during the 6 week treatment regimen.

[0057] Test population

[0058] This study is an open label trial, which aims to recruit 46 participants aged between 21 -45, with an equal number of males and females to control for any gender effects. Recruitment of 23 males and 23 females for a total sample of 46 was targeted. Previous studies investigating the health benefits of Eurycoma longifolia root extracts have primarily targeted elderly male populations. This study, which targets a younger population, would help to clarify if Eurycoma longifolia root extract is beneficial for younger adults, and if the same is able to be used to treat insomnia. Participants from this age group would mainly beworking adults ortertiary institution students who experience some non-clinical difficulties sleeping and may benefit from the Eurycoma longifolia root extract supplement. Participants will be recruited on a voluntary basis through an online form where they will indicate their interest and leave their contact details for a study team member to reach out to them.

[0059] Inclusion Criteria

[0060] healthy individuals who meet all of the following criteria:• Between 21 to 45 years old;• Educational level of at least O Levels;• Have trouble sleeping, defined by obtaining a score of 8 or higher on the Insomnia Severity Index (ISI);• Able to provide informed consent;• Non-smoking;• Agree to be alcohol-free for the duration of the trial (11 weeks total); and• May drink up to 2 cups of coffee (~200mg caffeine) before 12pm for the duration of the trial (11 weeks total).

[0061] Exclusion Criteria

[0062] Participants who meet any of the following criteria will be excluded from the study:• History of psychiatric illness;• History of cancer, heart disease, kidney disease, or weakened immune system;• History of alcohol or drug dependence;• History of known sleep apnea;• Regular intake of drugs or quasi-drugs for similar effects tested in this trial, including but not limited to eszopiclone (Lunesta), zolpidem (Ambien), zolpidem ER (Ambien CR), and zaleplon (Sonata), benzodiazepines, anti-depressants, melatonin, glycine, GABA, magnesium, L-tryptophan, and L- theanine;• Pregnant or breastfeeding (Female participants will acknowledge on the signed informed consent form that they are not pregnant or breastfeeding. Last menstrual cycle dates from the date of informed consent screening will also be noted down.);• Usage of hormonal contraceptives (Female participants will acknowledge on the signed informed consent form that they are not using hormonal contraceptives.);• At risk of developing food or drug allergies; and• Affected sleeping patterns due to external factors including but not limited to caregiving responsibilities, excessive environmental noise, and a snoring partner.

[0063] All medications (prescription and over the counter), vitamin and mineral supplements, and / or herbs taken by the participant were documented.

[0064] Treatment regimen

[0065] This is a prospective open label trial with a pre-test post-test design to evaluate the efficacy of the Eurycoma longifolia root extract as a sleep supplement. This includes a 1-week pre-intervention phase to establish baseline measures, 6-week intervention period to assess the effects of the Eurycoma longifolia root extract, and a post- intervention follow-up 4 weeks later.

[0066] Participants were recruited on a rolling basis over a period of 2 months. Each participant will follow the following sequence: a) Full consent taken over a secure videoconferencing platform and screening using ISI; b) Week 0: Pre-intervention phase and measurement of baseline (TO); c) Week 1 to 6: 6-week treatment trial period 1st Dose: At least 60 min before bed, latest 11 pm,. Subsequent Doses: 30-60 min before bed, latest 11 pm; d) with data collection at end of week 3 (T1) and end of week 6 (T2); e) Week 10: Post-intervention follow up at week 10 (T3).

[0067] This study is an open label trial where all participants received the Eurycoma longifolia root extract for 6 weeks.

[0068] During the 6 week treatment period, participants were monitored through:• Weekly online survey for participants to report any adverse side effects.• Physician’s assessment of adverse side effects at baseline, week 3 and week 6

[0069] In light of the Eurycoma longifolia root extract being traditionally used as an aphrodisiac, participants were also asked to report their libido levels at baseline and through the weekly online survey to monitor for any aphrodisiac effects.

[0070] Changes in body temperature were also be monitored daily using an oral thermometer. Participants were instructed to record their body temperature upon waking up in the body temperature log.

[0071] Measurements taken

[0072] For the first round of data collection (TO), participants completed the following assessments:• Insomnia Severity Index (subjective measure of sleep quality and insomnia symptoms; 5 items)• Epworth Sleepiness Scale (ESS) (subjective measure of daytime sleepiness; 8 items)• Inquisit Stroop Task and Digit Span (measures of cognitive performance, specifically executive functioning and working memory; ~2min (colour-word with keyboard responding) and ~15min respectively)• Profile of Mood States 2 Adult Short (POMS-2) (measure of mood; 35 items)• World Health Organisation Quality of Life (only Physical Capacity, Psychological, and Social Relationships domains; total of 44 items)• blood was drawn to assay for blood (plasma / serum) cortisol, serotonin, oxytocin, and testosterone. In addition, these foods: avocados, bananas, butternuts, cantaloupe, dates, eggplant, grapefruit, hickory nuts, honeydew melon, kiwifruit, melon, nuts, pecans, pineapple, plantains, plums, tomatoes, orwalnuts, were found to be high in serotonin. Therefore, to prevent an unexpected increase in the analysed blood biomarkers, participants were required to refrain from consuming these foods at least 48 hours before the blood draw.• Participants tracked their sleep using a wearable device (Fitbit Charge 5) for one week before starting their dosage period to resolve any technical issues and get used to the device, and then continue using the wearable device throughout the 6-week trial period and 4-week follow-up period every day and night to sleep (excluding bathroom / shower time, during which Fitbit will be charged). (Fitbit wearable data includes: 1 Sleep stages, 2 heart rate & HRV, breathing rate & SpC>2)• 72 hours before their first in-person visit (TO), participants were required to stop the intake of any drugs or quasi drugs for similar effects tested in this trial. During the 6-week dosage period, participants were required to complete a weekly online survey to report any adverse side effects or change in sexual function and to monitor compliance to the dosage schedule. Change in sexual function was to monitor any risk of inappropriate sexual behaviour e.g. harassment. Participants also indicated in the weekly online survey if they have started any new concomitant therapies since the commencement of the supplement trial. Participants also came back to the lab at the mid (week 3; T1) and end (week 6; T2) points of the 6-week trial for repeated rounds of data collections. The same assessments as at TO were made.• Due to the possibility of participants missing certain pre-planned follow-up study visits (T1 , T2, T3) because of personal / medical reasons, or due to the research team’s administrative issues; participants who were unable to attend the original pre-planned follow-up visit were allowed to reschedule their visits. However, the dates of the rescheduled visits was within a window period of 1 week (+7 days) after the specific dates for the original follow-up visits.• Participants were instructed to record their body temperature upon waking up in the body temperature log

[0073] 46 healthy participants aged 21-41 , with Insomnia Severity Index (ISI) score >8 took one 200mg EL extract capsule daily for 6 weeks and completed > 90% of dosage. Assessments including ISI, Inquisit Strooptask, Digitspan, WHO-QOL were done at pre-treatment (TO), during treatment 3 weeks (T1), 6 weeks (T2), and post treatment 10 weeks (T3). The final sample included 44 participants for data analysis as 2 female participants withdrew from the study one due to illness and the other due to a mild adverse reaction that included a fever and rash.

[0074] Differences for assessments between each timepoints and TO were assessed using Linear Mixed Model analysis with participants as a random effect. Demographic variables such as Age, Sex, Race, and Working Status were controlled.

[0075] ISI scores significantly reduced, mean difference (T1) = -4.17, 95% Cl (-5.48, -2.85), p<.001 , T2 = -6.30, 95% Cl (-7.63, -4.97), p<.001 , T3 = 5.05, 95% Cl (-6.38, -3.72), p<.001. QOL Thinking, learning, memory and concentration scores significantly increased, mean difference (T1) = 1 .72, 95% Cl (.94, 2.51), p<.001 , T2 = 2.22, 95% Cl (1.44, 3.01), p<.001 , T3 = 1.27, 95% Cl (.48, 2.06), p=.002. Insomnia scores predicted thinking scores at T2, B = -.274 and T3, B = -.315, p<.001 , with the regression model explaining 23.0% (R2= 23) and 37.2% (R2= 372) of the variance respectively.

[0076] Stroop task response latency (ms) significantly decreased, mean difference (T1) = -148.67, 95% Cl (-210.38, -86.95), p<.001 , T2 = -194.65, 95% Cl (-256.71 , -132.29), p< 001 , T3 = -227.78, 95% Cl (- 289.99, -165.57), p< 001. Backward Digitspan recall increased, mean difference (T2) = .82, 95% Cl (.44, 1.21), p<.001 , T3 = 1.0, 95% Cl (.61 , 1.38), p<.001. Psychological QOL significantly improved, mean difference (T1) = 1.0, 95% Cl (.52, 1.49), p<.001 , T2 = 1.30, 95% Cl (.81 , 1.78), p<.001.

[0077] The results all indicate that treatment with EL improved psychological well-being, sleep quality and consequently cognitive flexibility, working memory.

[0078] Safety measures

[0079] One of the study team members was responsible for the reporting of Unanticipated Problems Involving Risk to Subjects or Others (UPIRSTSO) events and Serious Adverse Events based on definitions in section 9.2 and section 9.3. Such events were identified through weekly telephone-based interview by the study team, monitoring and reporting from study team members during assessment sessions and selfreporting from trial participants.

[0080] Unexpected

[0081] In terms of nature, severity or frequency of the problem as described in the study documentation (eg: Protocol, Consent documents etc).

[0082] Related or possibly related to participation in the research

[0083] Possibly related means there is a reasonable possibility that the problem may have been caused by the procedures involved in the research; and

[0084] Risk of harm

[0085] Suggests that the research places participants or others at a greater risk of harm (including physical, psychological, economic, or social harm) than was previously known or recognized.

[0086] Data Analysis

[0087] Determination of Sample Size

[0088] A total of 46 participants were recruited. This was determined by power analysis using the G*Power software with the following parameters: 1) desired power: 0.8, statistical test: ANOVA, 3) alpha value: 0.05, 4) expected effect: small-medium 0.2. This generated a total sample size of N=36. We chose a sample size of N=46 to account for drop-outs. Indeed there were 2 drop-outs leaving a final sample size of N=44.

[0089] The data collected was computed into Microsoft Excel and SPSS software for analysis. A repeated-measures, within-subjects ANOVA was used to analyse the outcome measures. Linear Mixed Model analysis was later determined to be more appropriate for analysis.

[0090] [Table 1]: Final sample.

[0091] [Table 2]: demographic breakdown of subjects in trial

[0092] Overall subjective quality of life - Study Subjective Assessments

[0093] The present study used a variety of indicators to measure sleep, mood, and cognitive improvements.

[0094] Sleep Quality Indicators - INSOMNIA SEVERITY INDEX (ISI)

[0095] Changes in insomnia severity of all participants were measured using the 7-item Insomnia Severity Index (ISI) across all timepoints on a 5-point Likert scale. Participants were required to rate the severity of their insomnia problem(s) from 0 (no difficulties / problems) to 4 (very severe difficulties / problems). Participants were also asked to rate how their sleeping difficulties have impacted their functioning and how satisfied they were with their current sleep patterns. Lastly, participants were asked to rate how noticeable to others and distressed they were about their current sleeping problem. A participant’s total score may range from 0 to 28 with higher scores indicating greater levels of insomnia severity.

[0096] [Table 3]: questions asked in ISI ranked as 0 = None / Very satisfied / Not at all / Not at all interfering / noticeable; 1 = Mild / A little / Barely; 2 = Moderate / Somewhat; 3 = Severe / Much; and 4 = Very / Very dissatisfied / Very much interfering / noticeable.

[0097] A score from 0-7 is not clinically significant insomnia. A score from 8-14 is subthreshold insomnia. A score from 15-21 is clinical insomnia. A score from 22-28 is severe clinical insomnia.

[0098] [Table 4]:Total ISI Scores (controlled)[Table 4A],Mean0df Standard ErrorTO 17.09 37.70 2.09T1 12.93 37.61 2.09T2 10.79 37.67 2.09T3 12.04 37.67 2.09aCovariates appearing are evaluated at the following values: Age = 27.42[Table 4B],Mean Difference p» Confidence IntervalLower UpperTO T1 -4.17** <.001 -5.48 -2.85T2 -6.30** <.001 -7.63 -4.97T3 -5.05** <.001 -6.38 -3.72*pc.O5, **p<.001bNo adjustments used

[0099] ISI scores of all participants (n = 43) significantly decreased from TO to T1 , mean difference = - 4.17, Cl (-5.48, -2.85), p< 001 . TO to T2, mean difference = -6.30, Cl (-7.63, -4.97), p<.001 . TO to T3, mean difference = -5.05, Cl (-6.38, -3.72), p<.001 . ISI scores of male participants (n = 23) significantly decreased from TO to T1 , mean difference = -4.26, Cl (-5.88, -2.64), p<.001. TO to T2, mean difference = -5.61 , Cl (- 7.23, -3.99), p<.001. TO to T3, mean difference = -4.74, Cl (-6.36, -3.12), p< 001. ISI scores of female participants (n = 21) significantly decreased from TO to T1 , mean difference = -4.12, Cl (-6.29, -1.96), p<.001. TO to T2, mean difference = -7.12, Cl (-9.32, -4.92), p<.001 . TO to T3, mean difference = -5.45, Cl (-7.65, -3.25), pc.001 . ISI scores of students (n = 16) significantly decreased from TO to T1 , mean difference = -5.56, Cl (-7.61 , -3.51), pc.001. TO to T2, mean difference = -6.75, Cl (-8.80, -4.70), pc.001. TO to T3, mean difference = -5.13, Cl (-7.17, -3.08), pc.001. ISI scores of working adults (n = 27) significantly decreased from TO to T1 , mean difference = -3.33, Cl (-5.10, -1 .56), pc.001 . TO to T2, mean difference = - 5.97, Cl (-7.76, -4.18), pc.001 . TO to T3, mean difference = -5.16, Cl (-6.95, 3.37), pc.001 .

[0100] Sleep Quality Indicators - EPWORTH SLEEPINESS SCALE (ESS)

[0101] Changes in daytime sleepiness and wakefulness were measured using the 8-item Epworth Sleepiness Scale (ESS) across all timepoints on a 4-point Likert scale. Participants rated from 0 (would never doze) to 3 (high chance of dozing) how likely they were to fall asleep / doze off while engaging in 8 different activities. These 8 activities differ widely in their somnificity and when summed together give us an individual's average sleep propensity across different kinds of activities they might engage in daily. A person’s score may range from 0 to 24 with higher scores indicating greater levels of daytime sleepiness and conversely lower levels of wakefulness.

[0102] [Table 5]: questions asked in ESS ranked as 0 = would never doze; 1 = Slight chance of dozing; 2 = Moderate chance of dozing; and 3 = High chance of dozing.

[0103] A score from 0-5 is lower normal daytime sleepiness. A score from 6-10 is Higher normal daytime sleepiness. A score from 11-12 is mild daytime sleepiness. A score from 13-15 is moderate excessive daytime sleepiness. A score from 16-24 is servere excessive daytime sleepiness.[Table 6]: Total ESS Scores (controlled)[Table 6A.Mean® df Standard ErrorTO 11.29 37.32 1.72T1 8.42 37.32 1.72T2 7.57 37.26 1.72T3 8.18 37.35 1.72aCovariates appearing are evaluated at the following values: Age = 27.42[Table 6B],Mean Difference pbConfidence IntervalLower UpperTO T1 -2.86** < 001 -3.95 -1.78T2 -3.72** <.001 -4.81 -2.63T3 -3.11** <.001 -4.20 -2.01 p<.05, **p<.001bNo adjustments used

[0104] ESS scores of all participants (n = 43) significantly decreased from TO to T1 , mean difference = - 2.87, Cl (-3.95, -1 .78), p<.001 . TO to T2, mean difference = -3.72, Cl (-4.81 , -2.63), p< 001 . TO to T3, mean difference = -3.11 , Cl (-4.20, -2.01), p< 001. ESS scores of male participants (n = 23) significantly decreased from TO to T1 , mean difference = - 3.04, Cl (-4.50, -1 .59), p<.001 . TO to T2, mean difference = -3.26, Cl (-4.71 , -1.81), p<.001. TO to T3, mean difference = -2.74, Cl (-4.19, -1.29), p<.001. ESS scores of female participants (n = 23) significantly decreased from TO to T1 , mean difference = - 2.70, Cl (-4.36, - 1 .03), p = .002. TO to T2, mean difference = -4.22, Cl (-5.92, -2.53), p<.001 . TO to T3, mean difference = - 3.51 , Cl (-5.20, -1.82), p<.001. ESS scores of students (n = 16) significantly decreased from TO to T1 , mean difference = - 2.06, Cl (-4.01 , -.11), p = .039. TO to T2, mean difference = -3.38, Cl (-5.32, -1 .43), p = .001 . TO to T3, mean difference = -2.13, Cl (-4.07, -.18), p = .033. ESS scores of working adults (n = 27) significantly decreased from TO to T1 , mean difference = - 3.52, Cl (-4.85, -2.20), p<.001 . TO to T2, mean difference = -4.18, Cl (-5.32, -2.84), p< 001. TO to T3, mean difference = -3.74, Cl (-5.08, -2.39), p<.001.

[0105] Mood Indicators - PROFILE OF MOOD STATES 2 (POMS-2)

[0106] Changes in participants' mood and emotional states were measured using the 35-item Profile of Mood States 2 Adult Short (POMS-2) questionnaire across all timepoints. Participants rated on a 5-point Likert scale from 0 (Not at all) to 5 (Extremely) the intensity of each mood state that best described how they have “been feeling during the past week, including today”. Each item corresponds to a particular feeling such as anger, sadness or hopelessness which can then be further summed up into 7 different mood states (Anger-Hostility, Tension-Anxiety, Confusion-Bewilderment, Vigour-Activity, Fatigue-Inertia, Depression- Dejection, Friendliness). Total Mood Disturbance scores can also be calculated from the mood states (excluding Friendliness).

[0107] [Table 7] Summary of POMS-2

[0108] [Table 8]: POMS-2 Anger-Hostility (controlled)

[0109] [Table 8A],Mean3df Standard ErrorTO 4.65 39.93 1.18T1 3.85 39.93 1.18T2 3.17 40.03 1.19T3 3.39 40.03 1.19100110]aCovariates appearing are evaluated at the following values: Age = 27.42

[0111] [Table 8B].Mean pbhC „onfiden .Difference ce IntervalLower UpperTO T1 -.80 .11 -1.77 .17T2 -1.49* .003 -2.47 -.51T3 -1.26* .012 -2.24 -.28

[0112] *p< 05, **p<.001bNo adjustments used

[0113] [Table 91: POMS-2 Tension-Anxiety (controlled)[Table 9A],Mean3df Standard ErrorTO 5.46 37.36 1.70T1 4.90 37.36 1.70T2 4.13 37.41 1.70T3 3.86 37.41 1.703Covariates appearing are evaluated at the following values: Age = 27.42[Table 9B],Mean6Confidence IntervalDifferenceLower UpperTO T1 -.56 .27 -1.55 .44T2 -1.33* .009 -2.33 -.33T3 -1.60* .002 -2.60 -.60*pc.O5, **p<.001bNo adjustments used

[0114] Tension-Anxiety T-scores of all participants (n = 44) significantly decreased from TO to T2, mean difference = -4.16, Cl (-6.64, -1 .68), p = .001 . TO to T3, mean difference = -4.57, Cl (-7.05, -2.09), p<.001 . Tension-Anxiety T-scores of male participants (n = 23) significantly decreased from TO to T3, mean difference = -3.70, Cl (-6.77, -.63), p = .019. Tension-Anxiety T-scores of female participants (n = 22) significantly decreased from TO to T2, mean difference = -5.40, Cl (-9.47, -1 .33), p = .010. TO to T3, mean difference = -5.49, Cl (-9.57, -1 .42), p = .009. Tension-Anxiety T-scores of students (n = 16) significantly decreased from TO to T3, mean difference = -5.63, Cl (-10.41 , -.84), p = .022. Tension-Anxiety T-scores of working adults (n = 28) significantly decreased from TO to T2, mean difference = - 3.94, Cl (-6.86, -1 .02), p = .009. TO to T3, mean difference = -3.87, Cl (-6.79, -.95), p = .010.

[0115] [T able 101: POMS-2 Confusion-Bewilderment (controlled)[Table 10A],Mean3df Standard ErrorTO 7.86 36.76 1.74T1 6.15 36.76 1.74T2 5.91 36.80 1.74T3 5.79 36.80 1.743Covariates appearing are evaluated at the following values: Age = 27.42[Table 10b],Mean „ erence pbC „onfiden . . .Diff ce IntervalLower UpperTO T1 -1.71** < 001 -2.60 -.83T2 -1.95** < 001 -2.84 -1.06T3 -2.07** <.001 -2.96 -1.17*p<.05, **p< 001bNo adjustments used

[0116] [Table 11 ]: POMS-2 Depression-Dejection (controlled)[Table 11 A],TO 4.12 37.40 1.55T1 3.12 37.40 1.55T2 2.89 37.45 1.55T3 2.98 37.45 1.55aCovariates appearing are evaluated at the following values: Age = 27.42[Table 11 B],Mean pbhC „ .. . . . .Difference onfidence nterva*pc.O5, **p<.001bNo adjustments used

[0117] [T able 12]: POMS-2 Fatigue-Inertia (controlled)[Table 12A],Mean3df Standard ErrorTO 9.55 36.92 2.31T1 7.21 36.92 2.31T2 5.75 36.97 2.31T3 6.97 36.97 2.313Covariates appearing are evaluated at the following values: Age = 27.42[Table 12B],Meanh„c. . . .D..if„ference pbConfidence Interva .lLower UpperTO T1 -2.33** c.001 -3.56 -1.11T2 -3.80** c.001 -5.04 -2.57T3 -2.57** c.001 -3.81 -1.34*pc.O5, **p<.001bNo adjustments used

[0118] [T able 13]: POMS-2 Total Mood Disturbance (controlled)[Table 13A],Mean3df Standard ErrorTO 22.85 36.64 8.00T1 16.16 36.64 8.00T2 12.36 36.68 8.00T3 14.77 36.68 8.00aCovariates appearing are evaluated at the following values: Age = 27.42[Table 13B],Mean pb„ „ ,. . , , .Difference Confidence IntervalLower UpperTO T1 -6.69** c.001 -10.59 -2.78T2 -10.48** c.001 -14.42 -6.55T3 -8.07** c.001 -12.00 -4.14*pc.O5, **p<.001bNo adjustments used

[0119] Total Mood Disturbance T-scores of all participants (n = 44) significantly decreased from TO to T1 , mean difference = -4.16, Cl (-6.63, -1.68), p = .001 . TO to T2, mean difference = -6.99, Cl (-9.48, - 4.50), p<.001 . TO to T3, mean difference = -5.10, Cl (-7.60, -2.61), p<.001 . Total Mood Disturbance T- scores of male participants (n = 23) significantly decreased from TO to T1 , mean difference = -3.91 , Cl (- 7.22, -.61), p = .021 . TO to T2, mean difference = -6.04, Cl (-9.35, -2.74), pc.001 . TO to T3, mean difference = -4.04, Cl (-7.35, -.74), p = .017. Total Mood Disturbance T-scores of female participants (n = 22) significantly decreased from TO to T1 , mean difference = -4.41 , Cl (-8.23, -.59), p = .024. TO to T2, mean difference = -8.00, Cl (-11.88, -4.12), pc.001 . TO to T3, mean difference = -6.24, Cl (-10.1 1 , -2.36), p = .002. There were no significant differences in Total Mood Disturbance T-scores of students (n = 16) across all timepoints. Total Mood Disturbance T-scores of working adults (n = 28) significantly decreasedfrom TO to T1 , mean difference = -4.79, Cl (-7.39, -2.18), p<.001 . TO to T2, mean difference = -8.21 , Cl (- 10.85, -5.57), p<.001. TO to T3, mean difference = -6.14, Cl (-8.78, -3.50), p<.001.

[0120] POMS-2 Vigour-Activity T-Score

[0121] There were no significant changes in Vigour-Activity T-scores of all participants (n = 44) across all timepoints. There were no significant changes in Vigour-Activity T-scores of male participants (n = 23) across all timepoints. There were no significant changes in Vigour-Activity T-scores of female participants (n = 23) across all timepoints. There were no significant changes in Vigour-Activity T-scores of students (n = 16) across all timepoints. There were no significant changes in Vigour-Activity T-scores of working adults (n = 28) across all timepoints.

[0122] POMS-2 Friendliness T-Score

[0123] There were no significant changes in Friendliness T-scores of all participants (n = 44) across all timepoints. There were no significant changes in Friendliness T-scores of male participants (n = 23) across all timepoints. There were no significant changes in Friendliness T-scores of female participants (n = 22) across all timepoints. There were no significant changes in Friendliness T-scores of students (n = 16) across all timepoints. There were no significant changes in Friendliness T-scores of working adults (n = 28) across all timepoints.

[0124] Quality of Life Indicators - WHO Quality of Life-100 (WHO-QOL)

[0125] Changes in Physical, Psychological and Social Relationships quality of life domains were measured using the World Health Organisation Quality of Life (WHO-QOL), with only a total of 44 relevant items being included in the questionnaire administered to participants. Each domain consisted of a combination of facets such as Pain and Discomfort, Positive Thinking, Social Support etc. Each item was rated on a 5-point Likert scale ranging from 1 (Not at all) to 5 (Extremely). Domain scores are scaled such that higher scores denote a higher quality of life, while certain facets like Pain and Discomfort and Negative Feelings are scaled negatively such that lower scores denote a higher quality of life in that area.

[0126] [Table 14]: questions asked in WHO-QOL ranked as 0 = Not at all / Never / poor; 1 = A little / Slightly / A little / Poor / Seldom; 2 = Moderately / Neither poor nor good / Quite often; 3 = Mostly / Quite often / Good / Very Often; and 4 = Extremely / Very good / Completely / Always.

[0127] Higher scores indicate better QOL Except for Pain and Discomfort & Negative Feelings Facets.

[0128] [Table 15]: QOL Pain and Discomfort Facet (controlled)[Table 15A],Mean3df Standard ErrorTO 8.36 37.77 1.03T1 7.06 37.64 1.02T2 7.50 37.72 1.03T3 8.22 37.72 1.03aCovariates appearing are evaluated at the following values: Age = 27.42[Table 15B], ..„ Mean p6h„ Concf.id.ence . Int .e .Difference rvalLower UpperTO T1 -1.30* .002 -2.09 -.51T2 -.86* .035 -1.66 -.062T3 .13 .74 -.93 -.67*p<.05, **p<.001bNo adjustments used

[0129] Pain and Discomfort quality of life scores of all participants (n = 43) significantly decreased from TO to T1 , mean difference = - 1 .30, Cl (-2.09, -.51), p = .002. TO to T2, mean difference = -.86, Cl (- 1 .66, -.062), p = .035. Pain and Discomfort quality of life scores of male participants (n = 23) significantlydecreased from TO to T1 , mean difference = - 1.35, Cl (-2.39, -.30), p = .012. Pain and Discomfort quality of life scores of female participants (n = 21) significantly decreased from TO to T1 , mean difference = - 1.28, Cl (-2.48, -.08), p = .037. TO to T2, mean difference = -1 .35, Cl (-2.57, -.13), p = .031. Pain and Discomfort quality of life scores of students (n = 16) significantly decreased from TO to T1 , mean difference = - 1.38, Cl (-2.68, -.08), p = .039. Pain and Discomfort quality of life scores of working adults (n = 27) significantly decreased from TO to T1 , mean difference = - 1 .20, Cl (-2.25, -.14), p = .027.

[0130] [T able 16]: QOL Energy and Fatigue Facet (controlled)[Table 16A],TO 11.10 38.30 1.22T1 12.95 38.19 1.22T2 12.98 38.26 1.22T3 12.66 38.26 1.220Covariates appearing are evaluated at the following values: Age = 27.42[Table 16B],Mean ference phbC „onf ,.i . . . ,Dif dence IntervalTO T1 1.86** < 001 .99 2.72T2 1.88** <.001 1.01 2.76T3 1.56** <.001 .69 2.44*p<.05, **p<.001bNo adjustments used

[0131] Energy and Fatigue quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = 1 .86, Cl (.99, 2.72), p<.001 . TO to T2, mean difference = 1 .88, Cl (1 .01 , 2.76), p<.001 . TO to T3, mean difference = 1 .56, Cl (.69, 2.44), p<.001 . Energy and Fatigue quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 2.09, Cl (.83, 3.34), p = .001. TO to T2, mean difference = 2.17, Cl (.92, 3.43), p<.001 . TO to T3, mean difference = 1 .61 , Cl (.36, 2.86), p = .013. Energy and Fatigue quality of life scores of female participants (n = 21) significantly increased from TO to T1 , mean difference = 1 .62, Cl (.37, 2.86), p = .012. TO to T2, mean difference = 1.57, Cl (.31 , 2.83), p = .016. TO to T3, mean difference = 1 .52, Cl (.26, 2.79), p = .019. Energy and Fatigue quality of life scores of students (n = 16) significantly increased from TO to T2, mean difference = 2.38, Cl (.83, 3.92), p = .003. Energy and Fatigue quality of life scores of working adults (n = 27) significantly increased from TO to T1 , mean difference = 2.42, Cl (1.35, 3.50), p<.001 . TO to T2, mean difference = 1 .70, Cl (.61 , 2.78), p = .003. TO to T3, mean difference = 1 .77, Cl (.68, 2.86), p = .002.

[0132] [Table 17]: QOL Sleep and Rest Facet (controlled)[Table 17A],Mean® df _ Standard ErrorTO 8.84 37.90 1.38T1 12.19 37.80 1.38T2 12.91 37.86 1.38T3 11.86 37.86 1.38aCovariates appearing are evaluated at the following values: Age = 27.42[Table 17B],Mean pb„ C „onf ,.id.ence , In ,t ,Difference ervalLower UpperTO T1 3.35** <.001 2.47 4.22T2 4.06** c.001 3.18 4.95T3 3.02** c.001 2.14 3.90*pc.O5, **p<.001bNo adjustments used

[0133] Sleep and Rest quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = 3.35, Cl (2.47, 4.22), p<.001 . TO to T2, mean difference = 4.06, Cl (3.18, 4.95), p<.001 . TO to T3, mean difference = 3.02, Cl (2.14, 3.90), p<.001 . Sleep and Rest quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 3.57, Cl (2.30, 4.83), p<.001 . TO to T2, mean difference = 3.78, Cl (2.51 , 5.05), p<.001 . TO to T3, mean difference = 3.30, Cl (2.04, 4.57), p<.001 . Sleep and Rest quality of life scores of female participants (n = 21) significantly increased from TO to T1 , mean difference = 3.12, Cl (1.89, 4.36), pc.001 . TO to T2, mean difference = 4.38, Cl (3.12, 5.63), pc.001 . TO to T3, mean difference = 2.71 , Cl (1 .45, 3.97), pc.001 . Sleep and Rest quality of life scores of students (n = 16) significantly increased from TO to T1 , mean difference = 4.06, Cl (2.87, 5.26), pc.001 . TO to T2, mean difference = 5.56, Cl (4.37, 6.76), pc.001 . TO to T3, mean difference = 3.56, Cl (2.37, 4.76), pc.001. Sleep and Rest quality of life scores of working adults (n = 27) significantly increased from TO to T1 , mean difference = 2.86, Cl (1 .66, 4.07), pc.001 . TO to T2, mean difference = 3.02, Cl (1 .80, 4.24), pc.001 . TO to T3, mean difference = 2.69, Cl (1 .47, 3.91), pc.001 .

[0134] [Table 18]: QOL Positive Feelings Facet (controlled)[Table 18A],Mean3df Standard ErrorTO 11.88 36.76 1.36T1 12.69 36.70 1.36T2 13.33 36.74 1.36T3 12.70 36.74 1.36aCovariates appearing are evaluated at the following values: Age = 27.42[Table 18B],Mean pb„ C „onf ..id.ence , Int .e ,Difference rvalLower UpperTO T1 .81* .029 .086 1.53T2 1.45** < 001 .73 2.18T3 .82* .028 .090 1.54*p<.05, **p<.001bNo adjustments used

[0135] Positive Feelings quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = .81 , Cl (.086, 1 .53), p = .029. TO to T2, mean difference = 1 .45, Cl (.73, 2.18), pc.001 . TO to T3, mean difference = .82, Cl (.090, 1 .54), p = .029. Positive Feelings quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 1 .04, Cl (.023, 2.06), p = .045. TO to T2, mean difference = 1 .61 , Cl (.59, 2.63), p = .002. TO to T3, mean difference = .87, Cl (-.15, 1 .89), p = .093. Positive Feelings quality of life scores of female participants (n = 21) significantly increased from TO to T2, mean difference = 1 .29, Cl (.21 , 2.36), p = .02. There were no significant changes in Positive Feelings quality of life scores of students (n = 16).

[0136] [Table 19]: QOL Thinking, Learning Memory and Concentration Facet (controlled)[Table 19A],Mean3df Standard ErrorTO 9.37 38.06 1.15T1 11.09 37.96 1.14T2 11.59 38.02 1.15T3 10.64 38.02 1.15aCovariates appearing are evaluated at the following values: Age = 27.42[Table 19B],Mean nb„ C „ ,. . . . .Difference onfidence IntervalLower UpperTO T1 1.72** c.001 .94 2.51T2 2.22** c.001 1.44 3.01T3 1.27* .002 .48 2.06*pc.O5, **pc,001bNo adjustments used

[0137] Thinking, Learning, Memory and Concentration quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = 1 .72, Cl (.94, 2.51), p<.001 . TO to T2, mean difference = 2.22, Cl (1 .44, 3.01), p<.001 . TO to T3, mean difference = 1 .27, Cl (.48, 2.06), p = .002. Thinking, Learning, Memory and Concentration quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 2.13, Cl (1 .11 , 3.15), p<.001. TO to T2, mean difference = 2.30, Cl (1 .28, 3.33), p<.001 . TO to T3, mean difference = 1 .70, Cl (.68, 2.72), p = .001 . Thinking, Learning, Memory and Concentration quality of life scores of female participants (n = 21) significantly increased from TO to T1 , mean difference = 1 .30, Cl (.073, 2.53), p = .038. TO to T2, mean difference = 2.14, Cl (.90, 3.39), p = .001 . Thinking, Learning, Memory and Concentration quality of life scores of students (n = 16) significantly increased from TO to T2, mean difference = 2.50, Cl (1 .21 , 3.79), p<.001. Thinking, Learning, Memory and Concentration quality of life scores of working adults (n = 27) significantly increased from TO to T1 , mean difference = 2.10, Cl (1 .08, 3.12), pc.001 , TO to T2, mean difference = 2.03, Cl (.99, 3.07), pc.001 . TO to T3, mean difference = 1 .55, Cl (.51 , 2.59), p = .004.

[0138] [Table 20]: QOL Self-Esteem Facet (controlled)[Table 20A],Mean3df Standard ErrorTO 13.57 37.09 1.03T1 14.26 37.00 1.02T2 14.59 37.06 1.02T3 13.75 37.06 1.023Covariates appearing are evaluated at the following values: Age = 27.42[Table 20B],Mean C „onf ,.id.en , . ,Differencece IntervalLower UpperTO T1 .69* .033 .058 1.33T2 1.03* .002 .39 1.67T3 .19 .57 -.45 .83 p<.05, **p<.001bNo adjustments used

[0139] Self-Esteem quality of life scores of all participants (n = 43) significantly increased from TO to T 1 , mean difference = .69, Cl (.058, 1 .33), p = .033. TO to T2, mean difference = 1 .03, Cl (.39, 1 .67), p = .002. Self-Esteem quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 1 .35, Cl (.39, 2.31), p = .007. TO to T2, mean difference = 1 .48, Cl (.52, 2.44), p = .003. There were no significant changes in Self-Esteem quality of life scores of female participants (n = 21). Self-Esteem quality of life scores of students (n = 16) significantly increased from TO to T2, mean difference = 1.19, Cl (.003, 2.37), p = .049. Self-Esteem quality of life scores of working adults (n = 27) significantly increased from TO to T1 , mean difference = 1 .35, Cl (.39, 2.31), p = .007. TO to T2, mean difference = 1.48, Cl (.52, 2.44), p = .003.

[0140] [T able 21 ]: QOL Body Image and Appearance Facet (controlled)[Table 21 A],TO 13.73 36.84 1.19T1 14.52 36.77 1.19T2 14.52 36.82 1.19T3 14.29 36.82 1.19aCovariates appearing are evaluated at the following values: Age = 27.42[Table 21 B],Mean phbConfidenc .Difference e IntervalLower UpperTO T1 .80* .022 .12 1.46T2 .80* .023 .11 1.47T3 .56 1.04 -.12 1.24*p<.05, **p<.001bNo adjustments used

[0141] Body Image and Appearance quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = .79, Cl (.12, 1.46), p = .022. TO to T2, mean difference = .79, Cl (.11 , 1 .47), p = .023. Body Image and Appearance quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 1 .09, Cl (.21 , 1.96), p = .016. Body Image and Appearance quality of life scores of female participants (n = 21) significantly increased from TO to T2, mean difference = 1.19, Cl (.13, 2.25), p = .029. There were no significant differences in Body Image and Appearance quality of life scores of students (n = 16). Body Image and Appearance quality of life scores of working adults (n = 27) significantly increased from TO to T2, mean difference = 1 .22, Cl (.37, 2.06), p = .005. TO to T3, mean difference = .88, Cl (.04, 1 .73), p = .041 .

[0142] [Table 22] :QOL Negative Feelings Facet (controlled)[Table 22A],Mean® df Standard ErrorTO 9.20 38.30 .86T1 8.19 38.17 .86T2 8.18 38.26 .86T3 8.18 38.26 .86aCovariates appearing are evaluated at the following values: Age = 27.42[Table 22B],p6Confidence IntervalLower UpperTO T1 -1.01* .017 -1.83 -.18T2 -1.01* .017 -1.85 -.18T3 -1.01* .017 -1.85 -.18*pc.O5, **p<.001bNo adjustments used

[0143] Negative Feelings quality of life scores of all participants (n = 43) significantly decreased from TO to T1 , mean difference = -1 .01 , Cl (-1.83, -.18), p = .017. TO to T2, mean difference = -1 .01 , Cl (-1.85, -.18), p = .017. TO to T3, mean difference = - 1.01 , Cl (-1 .85, -.18), p = .017. There were no significant changes in Negative Feelings quality of life scores of male participants (n = 23). Negative Feelings quality of life scores of female participants (n = 21) significantly decreased from TO to T2, mean difference = - 1.48, Cl (-2.77, -.19), p = .026. TO to T3, mean difference = - 1.23, Cl (-3.10, -.52), p = .007. There were no significant changes in Negative Feelings quality of life scores of students (n =16) across all timepoints. Negative Feelings quality of life scores of working adults (n = 27) significantly decreased from TO to T2, mean difference = -1 .17, Cl (-2.77, -.09), p = .026. TO to T3, mean difference = - 1 .33, Cl (-2.45, -.27), p = .015.

[0144] [Table 23]: QOL Sexual Activity (controlled)[Table 23A],Mean3df Standard ErrorTO 13.91 36.09 1.38T1 14.73 36.09 1.38T2 15.26 36.07 1.38T3 14.78 36.07 1.38aCovariates appearing are evaluated at the following values: Age = 27.42[Table 23B],Mean pb„ C „onf ..id.ence . . .Difference IntervalLower UpperTO T1 .82* .009 .21 1.43T2 1.35** <.001 .73 1.97T3 .87* .006 .26 1.49 pc.05, **p< 001bNo adjustments used

[0145] Sexual Activity quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = .82, Cl (.21 , 1 .43), p = .009. TO to T2, mean difference = 1 .35, Cl (.73, 1.97), pc.001. TO to T3, mean difference = .87, Cl (.26, 1.49), p = .006. Sexual Activity quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 1 .22, Cl (.27, 2.16), p = .012. TO to T2, mean difference = 1 .30, Cl (.36, 2.25), p = .008. Sexual Activity quality of life scores of female participants (n = 21) significantly increased from TO to T2, mean difference = 1.41 , Cl (.63, 2.18), pc.001 . TO to T3, mean difference = 1 .12, Cl (.34, 1 .90), p = .005. Sexual Activity quality of life scores of students (n = 16) significantly increased from TO to T2, mean difference = 1.94, Cl (.63, 3.25), p = .005. TO to T3, mean difference = 1 .69, Cl (.38, 3.00), p = .013. Sexual Activity quality of life scores of working adults (n = 27) significantly increased from TO to T1 , mean difference = .70, Cl (.07, 1.33), p = .031. TO to T2, mean difference = 1.04, Cl (.40, 1.68), p = .002.

[0146] [Table 24]: QOL Physical Domain (controlled)[Table 24A],Mean3df Standard ErrorTO 11.86 37.31 .99T1 14.03 37.23 .99T2 14.13 37.28 .99T3 13.43 37.28 .993Covariates appearing are evaluated at the following values: Age = 27.42[Table 24B],Mean pbhC „onf ..id.en , , .Difference ce IntervalLower UpperTO T1 2.17** < 001 1.57 2.77T2 2.27** <.001 1.66 2.87T3 1 .57** <.001 .97 2.18*p<.05, **p<.001bNo adjustments used

[0147] Physical Domain quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = 2.17, Cl (1 .57, 2.77), p<.001. TO to T2, mean difference = 2.27, Cl (1 .66, 2.87), p<.001 . TO to T3, mean difference = 1 .57, Cl (.97, 2.18), p<.001 . Physical Domain quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 2.33, Cl (1.53, 3.1 ), p<.001 . TO to T2, mean difference = 2.13, Cl (1.33, 2.94), p<.001 . TO to T3, mean difference = 1.44, Cl (.63, 2.24), p<.001. Physical Domain quality of life scores of female participants (n = 21) significantly increased from TO to T1 , mean difference = 2.01 , Cl (1 .09, 2.93), p<.001 . TO to T2, mean difference = 2.43, Cl (1 .49, 3.37), p< 001 . TO to T3, mean difference = 1.73, Cl (.79, 2.67), p<.001. Physical Domain quality of life scores of students (n = 16) significantly increased from TO to T1 , mean difference = 2.17, Cl (1 .21 , 3.12), p< 001 . TO to T2, mean difference = 3.04, Cl (2.09, 4.00), p<.001 . TO to T3, mean difference =1 .67, Cl (.71 , 2.62), p = .001 . Physical Domain quality of life scores of working adults (n = 27) significantly increased from TO to T1 , mean difference = 2.16, Cl (1.37, 2.95), p<.001 . TO to T2, mean difference = 1 .75, Cl (.95, 2.55), p<.001 . TO to T3, mean difference =1 .49, Cl (.69, 2.29), p<.001.

[0148] [Table 25]: QOL Psychological Domain (controlled)[Table 25A],*p<.05, **p<.001bNo adjustments used

[0149] Psychological Domain quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = 1 .00, Cl (.52, 1 .49), p<.001 . TO to T2, mean difference = 1 .30, Cl (.81 , 1 .78), p< 001 . Psychological Domain quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = 1 .29, Cl (.64, 1.93), p<.001 . TO to T2, mean difference = 1 .29, Cl (.64, 1 .93), p<.001 . Psychological Domain quality of life scores of female participants (n = 21) significantly increased from TO to T2, mean difference = 1 .32, Cl (.57, 2.07), p< 001 . PsychologicalDomain quality of life scores of students (n = 16) significantly increased from TO to T2, mean difference = 1 .11 , Cl (.27, 1 .95), p = .011 . Psychological Domain quality of life scores of working adults (n = 27) significantly increased from TO to T1 , mean difference = 1 .15, Cl (.53, 1 .78), p< 001 . TO to T2, mean difference = 1.44, Cl (.81 , 2.07), p<.001.

[0150] [T able 26]: QOL Social Relationships Domain (controlled)[Table 26A],Mean3df Standard ErrorTO 14.25 36.13 1.16T1 14.77 36.09 1.16T2 14.90 36.11 1.16T3 14.65 36.11 1.16aCovariates appearing are evaluated at the following values: Age = 27.42[Table 26B],Mean phbConf ,.id .ence . In . .Difference tervalLower UpperTO T1 .53* .035 .038 1.01T2 .65* .010 .16 1.14T3 .40 .11 -.088 .89*p<.05, **p<.001bNo adjustments used

[0151] Social Relationships Domain quality of life scores of all participants (n = 43) significantly increased from TO to T1 , mean difference = .53, Cl (.039, 1 .01), p = .035. TO to T2, mean difference = .65, Cl (.16, 1.14), p = .010. Social Relationships Domain quality of life scores of male participants (n = 23) significantly increased from TO to T1 , mean difference = .88, Cl (.21 , 1.56), p = .011. Social Relationships Domain quality of life scores of female participants (n = 21) significantly increased from TO to T2, mean difference = .77, Cl (.048, 1 .49), p = .037. Social Relationships Domain quality of life scores of students (n = 16) significantly increased from TO to T2, mean difference = .88, Cl (.05, 1.70), p = .04. There were no significant differences in Social Relationships Domain quality of life scores of working adults (n = 27) across all timepoints.

[0152] WHO-QOL Social Support Facet

[0153] There were no significant changes in Social Support quality of life scores of all participants (n = 43) across all timepoints. There were no significant changes in Social Support quality of life scores of male participants (n = 23) across all timepoints. There were no significant changes in Social Support quality of life scores of female participants (n = 21) across all timepoints. There were no significant changes in Social Support quality of life scores of students (n = 16) across all timepoints. There were no significant changes in Social Support quality of life scores of working adults (n = 27) across all timepoints.

[0154] WHO-QOL Personal Relationships Facet

[0155] There were no significant changes in Personal Relationships quality of life scores of all participants (n = 43) across all timepoints. There were no significant changes in Personal Relationships quality of life scores of male participants (n = 23) across all timepoints. There were no significant changes in Personal Relationships quality of life scores of female participants (n = 21) across all timepoints. There were no significant changes in Personal Relationships quality of life scores of students (n = 16) across all timepoints. There were no significant changes in Personal Relationships quality of life scores of working adults (n = 27) across all timepoints.

[0156] Cognitive performance assessed using cognitive Assessments

[0157] All cognitive assessments were administered using the Inquisit Millisecond Software using a standardised research Samsung Galaxy Tablet.

[0158] Cognitive Flexibility - Stroop test

[0159] In a Stroop test the participants are asked cognitive questions for example: “In the following trials you will see words printed in different colours. Your task is to indicate the colour in which each word is printed while ignoring what the words actually say”. The responses are timed.

[0160] Improvements in Cognitive flexibility were derived by observing changes in participants’ Stroop Task mean response time and accuracy differences. Following the classic Stroop paradigm, participants were tasked to select the option which indicates the colour in which each word is printed while ignoring the actual meaning of the words. Participants were required to complete this task for both colour-meaning congruent and incongruent trials.

[0161] [Table 27]: colour stroop test

[0162] [T able 28]: Stroop Mean Response Time (RT) of All Correct Trials (controlled)[Table 28A],Mean3df Standard Error1375.25 36.38 137.881226.59 36.38 137.88T2 1180.75 36.41 137.91T3 1147.47 36.41 137.91aCovariates appearing are evaluated at the following values: Age = 27.42[Table 28B],Mean „ ference C „onf ,.id. , , .Dif ence IntervalLower UpperTO T1 -148.67** <.001 -210.38 -86.95T2 -194.50** <.001 -256.71 -132.29T3 -227.78** <.001 -289.99 -165.57*p<.05, **p<.001bNo adjustments used

[0163] Stroop Mean RT (ms) for all correct trials of all participants (n = 44) significantly decreased from TO to T1 , mean difference = - 148.67, Cl (-210.38, -86.95), p<.001. TO to T2, mean difference = - 194.50, Cl (-256.71 , -132.29), p<.001. TO to T3, mean difference = -227.78, Cl (-289.99, -165.57), pc.001 . Stroop Mean RT (ms) for all correct trials of male participants (n = 23) significantly decreased from TO to T1 , mean difference = - 161.80, Cl (-241.73, -81.87), p<.001. TO to T2, mean difference = - 213.35, Cl (-293.28, -133.42), p<.001. TO to T3, mean difference = -260.85, Cl (-340.78, -180.92), pc.001 . Stroop Mean RT (ms) for all correct trials of female participants (n = 22) significantly decreased from TO to T1 , mean difference = - 134.93, Cl (-232.61 , -37.25), p = .008. TO to T2, mean difference = - 174.23, Cl (-273.51 , -74.95), p<.001 . TO to T3, mean difference = -191 .94, Cl (-291 .22, -92.67), p<.001 . Stroop Mean RT (ms) for all correct trials of students (n = 16) significantly decreased from TO to T1 , mean difference = -139.48, Cl (-224.86, -54.09), p = .002. TO to T2, mean difference = -186.68, Cl (-272.06, - 101.30), p<.001. TO to T3, mean difference = -182.57, Cl (-267.96, -97.19), pc.001 . Stroop Mean RT (ms) for all correct trials of working adults (n = 28) significantly decreased from TO to T1 , mean difference = - 128.81 , Cl (-208.30, -49.32), p = .002. TO to T2, mean difference = -174.47, Cl (-255.01 , -93.92), pc.001. TO to T3, mean difference = -225.33, Cl (-305.87, -144.79), pc.001 .

[0164] [T able 291: Stroop Mean RT of Congruent Correct Trials (controlled)[Table 29A],Mean® df Standard ErrorTO 1339.13 37.35 118.82T1 1207.20 37.35 118.82T2 1121.46 37.40 118.87T3 1118.79 37.40 118.87aCovariates appearing are evaluated at the following values: Age = 27.42[Table 29B],Mean p1„1Conf ,.id .en . . .Difference ce ntervaLower UpperTO T1 -131.93** <.001 -201.16 -62.70T2 -217.67** <.001 -287.45 -147.90T3 -220.34** <.001 -290.12 -150.57*p<.05, **p<.001bNo adjustments used

[0165] Stroop Mean RT (ms) for all correct congruent trials of all participants (n = 44) significantly decreased from TO to T1 , mean difference = - 131.93, Cl (-238.54, -25.31), p=.O16. TO to T2, mean difference = -216.03, Cl (-323.25, -108.80), p< 001 . TO to T3, mean difference = -218.70, Cl (-325.92, - 111 .47), p<.001 . Stroop Mean RT (ms) for all correct congruent trials of male participants (n = 23) significantly decreased from TO to T1 , mean difference = -142.41 , Cl (-247.44, -37.38), p=.009. TO to T2, mean difference = -205.91 , Cl (-310.94, -100.88), p< 001 . TO to T3, mean difference = -220.29, Cl (- 325.32, -115.26), p<.001. Stroop Mean RT (ms) for all correct congruent trials of female participants (n = 22) significantly decreased from TO to T1 , mean difference = -120.97, Cl (-214.50, -27.43), p=.O12. TO to T2, mean difference = -230.34, Cl (-325.32, -135.35), p<.001. TO to T3, mean difference = -220.17, Cl (- 315.16, -125.19), p<.001. Stroop Mean RT (ms) for all correct congruent trials of students (n = 16) significantly decreased from TO to T1 , mean difference = -139.29, Cl (-254.80, -23.79), p=.O19. TO to T2, mean difference = -205.57, Cl (-321 .07, -90.06), p<.001 . TO to T3, mean difference = -151.33, Cl (- 266.83, -35.82), p = .011 . Stroop Mean RT (ms) for all correct congruent trials of working adults (n = 28) significantly decreased from TO to T1 , mean difference = -109.85, Cl (-196.33, -23.38), p=.O13. TO to T2, mean difference = -205.72, Cl (-293.32, -118.12), p<.001 . TO to T3, mean difference = -240.01 , Cl (- 327.61 , -152.41), p<.001.

[0166] [Table 30]: Stroop Mean RT of Incongruent Correct Trials (controlled)[Table 30A],Mean3df Standard ErrorTO 1561.09 36.87 194.29T1 1430.43 36.87 194.29T2 1352.43 36.91 194.35T3 1280.09 36.91 194.35aCovariates appearing are evaluated at the following values: Age = 27.42[Table 30B],Mean ence phbC „onfide ,Differ nce IntervalLower UpperTO T1 -130.66** .011 -231.38 -29.94T2 -208.66** <.001 -310.18 -107.13T3 -281.00* <.001 -382.52 -179.47*p<.05, **p<.001bNo adjustments used

[0167] Stroop Mean RT (ms) for all correct incongruent trials of all participants (n = 44) significantly decreased from TO to T2, mean difference = -208.17, Cl (-377.44, -38.89), p=.O16. TO to T3, mean difference = -280.51 , Cl (-449.78, -11 1 .23), p = .001 . Stroop Mean RT (ms) for all correct incongruenttrials of male participants (n = 23) significantly decreased from TO to T2, mean difference = -237.25, Cl (- 378.21 , -96.29), p=.001. TO to T3, mean difference = -328.39, Cl (-469.35, -187.43), p <.001. Stroop Mean RT (ms) for all correct incongruent trials of female participants (n = 22) significantly decreased from TO to T2, mean difference = -177.24, Cl (-328.42, 26.06), p= .001 . TO to T3, mean difference = -229.00, Cl (-380.17, -77.82), p = .004. Stroop Mean RT (ms) for all correct incongruent trials of students (n = 16) significantly decreased from TO to T1 , mean difference = 169.29, Cl (-290.76, -47.81), p=.007. TO to T2, mean difference = -173.66, Cl (-295.13, -52.18), p = .006. TO to T3, mean difference = -237.46, Cl (- 358.94, -115.99), p<.001 , Stroop Mean RT (ms) for all correct incongruent trials of working adults (n = 28) significantly decreased from TO to T2, mean difference = -194.64, Cl (-332.18, -57.10), p = .006. TO to T3, mean difference = -261 .54, Cl (-399.08, -124.00), p<.001 ,

[0168] Stroop Task: Overall Proportion of Correct Trials

[0169] There were no significant differences in Stroop overall proportion of correct trials of all participants (n = 44) across all timepoints. Stroop overall proportion of correct trials of male participants (n = 23) significantly decreased from TO to T1 , mean difference = .013, Cl (.002, .024), p = .026. There were no significant differences in Stroop overall proportion of correct trials of females (n = 22) across all timepoints. There were no significant differences in Stroop overall proportion of correct trials of students (n = 16) across all timepoints. There were no significant differences in Stroop overall proportion of correct trials of working adults (n = 28) across all timepoints.

[0170] [T able 31 ]: Stroop Proportion of Correct Congruent Trial (controlled)[Table 31 A].Mean3df Standard ErrorTO .992 41.23 .01T1 .982 41.23 .01T2 .986 41.36 .01T3 .984 41.36 .01aCovariates appearing are evaluated at the following values: Age = 27.42[Table 31 B],Mean pb„ C „on .Difference fidence IntervalLower UpperTO T1 -.01* .03 -.02 -.001T2 -.006 .21 -.015 .003T3 -.008 .11 -.017 .002*p<.05, **p< 001bNo adjustments used

[0171] Stroop proportion of correct congruent trials of all participants (n = 44) significantly decreased from TO to T1 , mean difference = -.010, Cl (-.020, -.001), p = .030. There were no significant differences in Stroop proportion of correct congruent trials of male participants (n = 23) across all timepoints. Stroopproportion of correct congruent trials of female participants (n = 23) significantly decreased from TO to T1 , mean difference = -.016, Cl (-.032, -.001), p = .041. Stroop proportion of correct congruent trials of students (n = 16) significantly decreased from TO to T1 , mean difference = -.022, Cl (-.040, -.004), p = .015. TO to T3, mean difference = -.020, Cl (-.038, -.002), p = .028. There were no significant differences in Stroop proportion of correct congruent trials of working adults (n = 28) across all timepoints.

[0172] [T able 32]: Stroop Proportion of Incongruent (controlled)[Table 32A],Mean® df Standard ErrorTO .941 42.67 .018T1 .957 42.67 .018T2 .955 42.83 .018T3 .957 42.83 .018aCovariates appearing are evaluated at the following values: Age = 27.42[Table 32B],Confidence IntervalLower UpperTO T1 .016 .093 -.003 .034T2 .014 .14 -.005 .033T3 .017 .08 -.002 .035 pc.05, **p< 001bNo adjustments used

[0173] There were no significant differences in Stroop proportion of correct incongruent trials of all participants (n = 44) across all timepoints. Stroop proportion of correct incongruent trials of male participants (n = 23) significantly decreased from TO to T1 , mean difference = .043, Cl (.016, .071), p = .002. TO to T2, mean difference = .034, Cl (.007, 061), p = .015. There were no significant differences in Stroop proportion of correct incongruent trials of female participants (n = 22) across all timepoints. There were no significant differences in Stroop proportion of correct incongruent trials of students (n = 16) across all timepoints. There were no significant differences in Stroop proportion of correct incongruent trials of working adults (n = 28) across all timepoints.

[0174] Working Memory - Digit span assessment

[0175] The participants are tasked to remember number sequences presented. The cognitive performance task, Digit Span (Forward and Backward recall), was administered whereby participants had to recall digit sequences in either a forward or backward manner. Depending on their performance, participants either move up ordown a level. A progression in task levels indicate that participants can recall the digits in sequence accurately and therefore reflected higher working memory abilities.

[0176] [Table 33]: VISUAL DIGIT SPAN TEST

[0177] DigitSpan Task: Traditional Forward Measure

[0178] There were no significant differences in DigitSpan traditional forward measure of all participants (n = 44) across all timepoints. DigitSpan traditional forward measure of male participants (n = 23) significantly increased from TO to T3, mean difference = 1 .61 , Cl (.24, 2.97), p=.O22. There were no significant changes in DigitSpan traditional forward measure of female participants (n = 23). There were no significant changes in DigitSpan traditional forward measure of students (n = 16). DigitSpan traditional forward measure of working adults (n = 28) significantly increased from TO to T3, mean difference = 1.61 , Cl (.24, 2.97), p=.022.

[0179] [T able 341: DigitSpan Traditional Backward Measure (controlled)[Table 34A],Mean® df Standard ErrorTO 6.73 39.57 .71T1 7.04 39.57 .71T2 7.33 39.67 .71T3 7.72 39.67 .71aCovariates appearing are evaluated at the following values: Age = 27.42[Table 34B],MeanConfidence IntervalDifferenceLower UpperTO T1 .31 .28 -.26 .88T2 .60* .04 .028 1.18T3 .99** <.001 .41 1.56*pc.O5, **p<.001bNo adjustments used

[0180] DigitSpan traditional backward measure of all participants (n = 44) significantly increasedfrom TO to T3, mean difference = .98, Cl (.25, 1.70), p=.009. Digitspan traditional backward measure of male participants (n = 23) significantly increased from TO to T3, mean difference = 1 .44, Cl (.54, 2.33), p=.002. There were no significant changes in Digitspan traditional backward measure of female participants (n = 22). There were no significant changes in Digitspan traditional backward measure of students (n = 16). Digitspan traditional backward measure of working adults (n = 28) significantly increased from TO to T3, mean difference = 1 .13, Cl (.33, 1.92), p=.006.

[0181] Digitspan Task: Maximal Forward Measure

[0182] There were no significant changes in Digitspan maximal forward measure of all participants (n = 44). There were no significant changes in Digitspan maximal forward measure of male participants (n = 23), female participants (n = 22), and students (n = 16). Digitspan maximal forward measure of working adults (n = 28) significantly increased from TO to T1 , mean difference = .93, Cl (.14, 1.72), p = .022.

[0183] [Table 35]: DiqitSpan Maximal Backward Measure (controlled)[Table 35A],Mean3df Standard ErrorTO 7.28 36.99 .70T1 7.54 36.99 .70T2 8.07 37.03 .70T3 8.09 37.03 .70aCovariates appearing are evaluated at the following values: Age = 27.42[Table 35B],Mean ce pb„ C „onfidenc ,Differen e IntervalLower UpperTO T1 .27 .16 -.11 .64T2 .79** <.001 .42 1.17T3 .81** <.001 .44 1.19*p<.05, **p<.001bNo adjustments used

[0184] DigitSpan maximal backward measure of all participants (n = 44) significantly increased from T0 to T2, mean difference = .79, Cl (.17, 1.40), p=.012. TO to T3, mean difference = .81 , Cl (.19, 1.42), p = .01. DigitSpan maximal backward measure of male participants (n = 23) significantly increased from TO to T1 , mean difference = .52, Cl (.037, 1.01), p=.O35. TO to T2, mean difference = .87, Cl (.39, 1 .35), p<.001 . TO to T3, mean difference = .83, Cl (.34, 1 .31), p = .001 . DigitSpan maximal backward measure of female participants (n = 22) significantly increased TO to T2, mean difference = .71 , Cl (.11 , 1.30), p = .02. TO to T3, mean difference = .80, Cl (.21 , 1 .39), p = .009. DigitSpan maximal backward measure of students (n = 16) significantly increased TO to T2, mean difference = .69, Cl (.063, 1.31), p = .032. TO to T3, mean difference = .69, Cl (.063, 1.31), p = .032. DigitSpan maximal backward measure of working adults (n = 28) significantly increased TO to T2, mean difference = .84, Cl (.34, 1 .35), p = .001 . TO to T3, mean difference = .88, Cl (.38, 1.38), p<.001.

[0185] Digitspan Task: New Forward Measure

[0186] There were no significant differences in Digitspan new forward measure of all participants (n = 44) across all timepoints. Digitspan new forward measure of male participants (n = 23) significantly increased fromTO to T3, mean difference = 1.45, Cl (.33, 2.56), p = .012. There were no significant changes in Digitspan new forward measure of female participants (n = 23) across all timepoints. There were no significant changes in Digitspan new forward measure of students (n = 16) across all timepoints. There were no significant changes in Digitspan new forward measure of working adults (n = 28) across all timepoints.

[0187] [Table 36]: DiqitSpan New Backward Measure (controlled)[Table 36A],Mean3df Standard ErrorTO 6.62 37.06 .70T1 6.94 37.06 .70T2 7.44 37.11 .70T3 7.62 37.11 .70aCovariates appearing are evaluated at the following values: Age = 27.42[Table 36B],Meanh„ .Difference pbConfidence IntervalLower UpperTO T1 .33 .092 -.054 .71 <.001 .44 1.21<.001 .61 1.38*p<.05, **p<.001bNo adjustments used

[0188] DigitSpan new backward measure of all participants (n = 44) significantly increased from TO to T2, mean difference = .82, Cl (.20, 1 .43), p=.01 . TO to T3, mean difference = .99, Cl (.37, 1 .61), p = .002. DigitSpan new backward measure of male participants (n = 23) significantly increased from TO to T 1 , mean difference = .53, Cl (.001 , 1 .05), p= .01 . TO to T2, mean difference = .82, Cl (.30, 1 .35), p = .003. TO to T3, mean difference = .89, Cl (.37, 1 .42), p = .001 . DigitSpan new backward measure of female participants (n = 22) significantly increased from TO to T2, mean difference = .82, Cl (.25, 1 .40), p=.006. TO to T3, mean difference = 1.11 , Cl (.54, 1.68), p<.001. DigitSpan new backward measure of students (n = 16) significantly increased from TO to T2, mean difference = .69, Cl (.11 , 1 .28), p=.O22. TO to T3, mean difference = .65, Cl (.06, 1 .23), p = .031. DigitSpan new backward measure of working adults (n = 28) significantly increased from TO to T2, mean difference = .91 , Cl (.38, 1 .44), p<.001 . TO to T3, mean difference = 1 .22, Cl (.69, 1 .74), p<.001 .

[0189] Summary of analysis

[0190] [Table 37][Table 37A]: Summary of preliminary analysis[Table 37B]: Summary of the subjective measurements. (To analyse for changes in the different variables from baseline to mid, post and post-cessation of IP, a Linear Mixed Model Analysis was conducted for all assessments so as to control for factors such as age, sex, race, working status and participant random effects)

[0191] The present study found that across all participants, improvements in sleep indicators were observed after the intake of Tongkat Ali. Participants’ subjective insomnia severity levels (ISI) significantly improved after the intake of Tongkat Ali. Participants’ subjective levels of insomnia were found to decrease significantly when comparing to baseline regardless of gender and working status. Participants’ subjective levels of daytime sleepiness (ESS) were also found to decrease across all timepoints when comparing to baseline, regardless of gender and working status.

[0192] Subjective changes in participants’ mood profile were also observed afterthe intake of Tongkat Ali. More specifically, participants reported a significant decrease in POMS-2 Anger-Hostility, Depression- Dejection, Fatigue-Inertia and Tension-Anxiety. Total mood disturbance also significantly decreased across all timepoints for all participants. This improvement in mood may have been influenced by participants’ sleep improvements.

[0193] The Eurycoma longifolia root extract reduced difficulties sleeping and promote sleep quality and its associated outcomes of cognition, mood, and overall quality of life. In addition to treating insomnia, Eurycoma longifolia root extract improved not just sleep but also its associated outcomes of cognition, mood, and overall quality of life.

[0194] Blood Biomarkers

[0195] changes in blood biomarkers

[0196] The biomarkers Oxytocin, Serotonin and Cortisol (all participants) testosterone (male participants only and 6 females with reported period abnormalities / increased libido) were all measured 1 - week pre-treatment phase, after 6-week treatment period, and 4 weeks post-treatment. Blood biomarkers were drawn from participants in the morning (within the hours of 8.30am to 12pm latest). Samples were iced immediately upon extraction and processed within 3 hours. Blood plasma was subsequently extracted and analysed using ELK Biotechnology ELISA kits in NUS’ Immunology Lab.

[0197] Serotonin

[0198] Serotonin, 5-hydroxtryptamine (5-HT) is a neurotransmitter and hormone that help to regulate one’s mood and sleep (Jones et aL, 2020). It was found in various studies to play a key role in the sleepwake cycle. It is also an integral neurotransmitter in many psychiatric disorders such as depression that result in hypersomnia or insomnia (Bamalan et al., 2023). Changes in serotonin levels were thus studied in order to better understand Tongkat Ali’s effect on mood and sleep.

[0199] There were no significant differences in Serotonin levels of all participants (n = 45) across all timepoints. There were no significant differences in Serotonin levels of male participants (n = 23) across all timepoints. There were no significant differences in Serotonin levels of female participants (n = 22) across all timepoints. There were no significant differences in Serotonin levels of students (n =16) across all timepoints. There were no significant differences in Serotonin levels of working adults (n = 28) across all timepoints.

[0200] Cortisol

[0201] Cortisol, a hormone secreted by the adrenal gland, plays a key role in one’s stress, mood,sleep and circadian rhythm. Elevated cortisol levels are known to result in an increase of sleep disturbances, mood disorders and stress-related issues. Other clinical studies have investigated the effects of other adaptogens such as Magnolia Officinalis and Ashwagandha on cortisol and have found optimistic findings (Bush, 2014). Changes in participants’ cortisol levels were thus studied to better understand the effects of Tongkat Ali on sleep.

[0202] Cortisol levels of all participants (n = 45) significantly increased from TO to T1 , mean difference = 26.82 Cl (10.31 , 43.32), p = .002. TO to T3, mean difference = 18.41 , Cl (1.79, 35.03), p = .03. Cortisol levels of male participants (n = 23) significantly increased from TO to T1 , mean difference = 39.57, Cl (13.03, 66.12), p = .004. There were no significant differences in Cortisol levels of female participants (n = 22) across all timepoints. Cortisol levels of students (n=16) significantly increased from TO to T1 , mean difference = 39.79, Cl (16.57, 63.02), p = .001. TO to T2, mean difference = 24.24, Cl (1.02, 47.47), p = .041 . TO to T3, mean difference = 25.15, Cl (1 .93, 48.37), p = .034. There were no significant differences in Cortisol levels of working adults (n = 28) across all timepoints.

[0203] Cortisol levels were found to increase after 3 weeks of Tongkat Ali and subsequently maintained 4-weeks post cessation of dosage for all participants. However, this may have been attributable to the study data collection period. Baseline measurements were collected during the start of the university semester while subsequent measurements were collected around the mid-term (T1) and final examinations period (T3). All students (n=16) that were enrolled in this study were also university students completing their tertiary education. Given that cortisol levels tend to increase during high periods of stress, the increase of cortisol levels during T1 and T3 may not have been due to the effects of Tongkat Ali, but may be from external confounders such as participants’ level of daily stress. This was further supported by a split analysis indicating that only students’ levels of cortisol were found to significantly increase while working adults did not experience any significant changes.

[0204] Oxytocin

[0205] Oxytocin, also known as the ‘love’ (cuddle) hormone, is a hypothalamic neuropeptide which influences different neurobiological and social processes. A study has found that oxytocin seems to promote restfulness while one is conscious (Raymond et al., 2021). Other research has found clinical possibilities of oxytocin-based interventions in influencing sleep-wake and sleep related outcomes. As such, this study examined the effects of Tongkat Ali on Oxytocin and its possible related effects to sleep and mood.

[0206] There were no significant differences in Oxytocin levels of all participants (n = 45) across all timepoints. Oxytocin level of male participants (n=23) significantly increased from TO to T1 , mean difference = 14.09, Cl (6.26, 21.92), p<.001. TO to T2, mean difference = 11.33, Cl (3.50, 19.16), p = .005. TO to T3, mean difference = 12.53, Cl (4.70, 20.36), p = .002. Oxytocin level of female participants (n=22) significantly decreased from TO to T1 , mean difference = -17.12, Cl (-29.93, -4.31), p = .01 . There were no significant differences in Oxytocin levels of students (n = 16) across all timepoints. There were no significant differences in Oxytocin levels of working adults (n = 28) across all timepoints.

[0207] Testosterone

[0208] Tongkat Ali has been marketed as an all-natural alternative to Hormone Replacement Therapy to address symptoms of aging that are associated with low testosterone levels. Previous studiesinvestigating its use have found a consistent effect on testosterone levels and sexual function. These studies however targeted older populations with infertility, hypogonadism or erectile dysfunction symptoms, suggesting that Tongkat Ali increases libido only in those with subnormal testosterone levels, while the libido of healthy individuals would simply be maintained (Chen et al., 2014, Kotirum et al., 2015). In this study, testosterone levels were measured for male participants for monitoring purpose.

[0209] There were no significant differences in Testosterone levels of all participants (n = 29) across all timepoints. Testosterone level of male participants (n = 23) significantly increased from TO to T2, mean difference = 39.64, Cl (20.51 , 58.77), p<.001 . TO to T3, mean difference = 27.14, Cl (5.96, 48.32), p = .013. Testosterone level of female participants (n = 6) significantly decreased from TO to T1 , mean difference = -88.82, Cl (-158.24, -19.39), p = .017. TO to T2, mean difference = -114.53, Cl (-183.96, - 45.11), p = .004. There were no significant differences in Testosterone levels of students (n = 11) across all timepoints. Testosterone level of working adults (n = 17) significantly increased from TO to T2, mean difference = 32.20, Cl (2.82, 61.58), p = .032.

[0210] Table 38: Summary of blood biomarker assessments.

[0211] Fitbit Charges

[0212] Each participant was also issued a Fitbit Charge 5 wearable device as an objective measure of sleep activity. Participants were required to wear the Fitbit Charge 5 one week before the start of their dosage period to measure their baseline levels of sleep activity. They were then instructed to continue using the wearable device throughout the 6-week trial / dosage period and during the 4-week follow-up period every day and night to sleep. Participants were allowed to remove the wearable device during bathroom / shower time to charge the Fitbit.

[0213] Fitbit: Total Minutes Asleep

[0214] There were no significant changes in Total number of Minutes Asleep of all participants (n = 44) across all timepoints. Total number of Minutes Asleep of male participants (n = 22) significantly decreased from TO to T3, mean difference = -23.93, Cl (-42.71 , -5.15), p = .013. There were no significantchanges in Total number of Minutes Asleep of female participants (n = 22) across all timepoints. There were no significant changes in Total number of Minutes Asleep of students (n = 16) across all timepoints. There were no significant changes in Total number of Minutes Asleep of working adults (n = 28) across all timepoints.

[0215] Fitbit: Sleep Latency

[0216] There were no significant changes in Sleep Latency of all participants (n = 44) across all timepoints. There were no significant changes in Sleep Latency of male participants (n = 22) across all timepoints. There were no significant changes in Sleep Latency of female participants (n = 22) across all timepoints. There were no significant changes in Sleep Latency of students (n = 16) across all timepoints. There were no significant changes in Sleep Latency of working adults (n = 28) across all timepoints.

[0217] Fitbit: Total number of minutes in Light Sleep

[0218] There were no significant changes in total number of minutes in light sleep of all participants (n = 44) across all timepoints. There were no significant changes in total number of minutes in light sleep of male participants (n = 22) across all timepoints. There were no significant changes in total number of minutes in light sleep of female participants (n = 22) across all timepoints. There were no significant changes in total number of minutes in light sleep of students (n = 16) across all timepoints. There were no significant changes in total number of minutes in light sleep of working adults (n = 28) across all timepoints.

[0219] Fitbit: Total number of minutes in Deep Sleep

[0220] There were no significant changes in total number of minutes in deep sleep of all participants (n = 44) across all timepoints. There were no significant changes in total number of minutes in deep sleep of male participants (n = 22) across all timepoints. There were no significant changes in total number of minutes in deep sleep of female participants (n = 22) across all timepoints. There were no significant changes in total number of minutes in deep sleep of students (n = 16) across all timepoints. There were no significant changes in total number of minutes in deep sleep of working adults (n = 28) across all timepoints.

[0221] Fitbit: Total number of minutes in REM Sleep

[0222] Total number of minutes in REM sleep of all participants (n = 44) significantly increased from TO to T2, mean difference = 6.15, Cl (.90, 11.41), p = .022. Total number of minutes in REM sleep of male participants (n = 22) significantly increased from TO to T2, mean difference = 8.83, Cl (1.39, 16.27), There were no significant changes in Total number of minutes in REM sleep of female participants (n = 22) across all timepoints. There were no significant changes in Total number of minutes in REM sleep of students (n = 16) across all timepoints. Total number of minutes in REM sleep of working adults (n = 28) significantly increased from TO to T1 , mean difference = 6.12, Cl (.48, 11 .76), p = .034. TO to T2, mean difference = 6.51 , Cl (.72, 12.30), p = .028.

[0223] Fitbit: Light Sleep, 30-Day Average

[0224] 30-day average of time spent in light sleep (per day) of all participants (n = 44) significantly increased from TO to T1 , mean difference = 38.67, Cl (30.21 , 47.13), p<.001 . TO to T2, mean difference = 36.30, Cl (27.63, 44.98), p<.001. TO to T3, mean difference = 34.76, Cl (26.02, 43.51), p<.001 .30-day average of time spent in light sleep (per day) of male participants (n = 22) significantly increased from TOto T1 , mean difference = 43.19, Cl (30.86, 55.51), pc.001. TO to T2, mean difference = 41 .22, Cl (28.90, 53.54), p<.001. TO to T3, mean difference = 37.77, Cl (25.26, 50.28), p<.001 . 30-day average of time spent in light sleep (per day) of fema30-dayticipants (n = 22) significantly increased from TO to T1 , mean difference = 34.14, Cl (22.19, 46.09), p<.001. TO to T2, mean difference = 30.96, Cl (18.36, 43.56), pc.001. TO to T3, mean difference = 31 .73, Cl (19.12, 44.33), pc.001 . 30-day average of time spent in light sleep (per day) of students (n = 16) significantly increased from TO to T1 , mean difference = 34.41 , Cl (17.62, 51.20), pc.001. T0 to T2, mean difference = 29.74, Cl (12.56, 46.92), p = .001. TO to T3, mean difference = 31.22, Cl (13.63, 48.81), pc.001. 30-day average of time spent in light sleep (per day) of working adults (n = 28) significantly increased from TO to T1 , mean difference = 40.06, Cl (30.05, 50.06), pc.001 . TO to T2, mean difference = 39.03, Cl (28.75, 49.31), pc.001 . TO to T3, mean difference = 35.21 , Cl (24.94, 45.49), pc.001.

[0225] Fitbit: Deep Sleep, 30 Day Average

[0226] 30-day average of time spent in deep sleep (per day) of all participants (n = 44) significantly increased from TO to T1 , mean difference = 11.70, Cl (8.68, 14.72), pc.001. TO to T2, mean difference = 11.07, Cl (7.97, 14.17), pc.001 . TO to T3, mean difference = 9.96, Cl (6.84, 13.08), pc.001 . 30-day average of time spent in deep sleep (per day) of male participants (n = 22) significantly increased from TO to T1 , mean difference = 13.59, Cl (10.07, 17.10), pc.001 . TO to T2, mean difference = 13.38, Cl (9.86, 16.90), pc.001. TO to T3, mean difference = 11.89, Cl (8.31 , 15.46), pc.001. 30-day average of time spent in deep sleep (per day) of female participants (n = 22) significantly increased from TO to T1 , mean difference = 9.78, Cl (4.70, 14.86), pc.001. TO to T2, mean difference = 8.63, Cl (3.28, 13.98), p = .002. TO to T3, mean difference = 8.01 , Cl (2.66, 13.35), p = .004. 30-day average of time spent in deep sleep (per day) of students (n = 16) significantly increased from TO to T1 , mean difference = 11.89, Cl (6.60, 17.18), pc.001. TO to T2, mean difference = 11.96, Cl (6.52, 17.34), pc.001. TO to T3, mean difference = 10.86, Cl (5.32, 16.40), pc.001. 30-day average of time spent in deep sleep (per day) of work30-dayults (n = 28) significantly increased from TO to T1 , mean difference = 11 .63, Cl (7.69, 15.57), pc.001 . TO to T2, mean difference = 10.43, Cl (6.39, 14.48), pc.001 . TO to T3, mean difference = 9.40, Cl (5.35, 13.45), pc.001.

[0227] Fitbit: REM Sleep, 30-Day Average

[0228] 30-day average of time spent in REM sleep (per day) of all participants (n = 44) significantly increased from TO to T1 , mean difference = 13.78, Cl (9.18, 18.38), pc.001. TO to T2, mean difference = 14.52, Cl (9.81 , 19.23), pc.001 . TO to T3, mean difference = 13.42, Cl (8.67, 18.17), pc.001 . 30-day average of time spent in REM sleep (per day) of male participants (n = 22) significantly increased from TO to T1 , mean difference = 13.78, Cl (7.78, 19.78), pc.001 . TO to T2, mean difference = 17.13, Cl (11 .13, 23.14), pc.001. TO to T3, mean difference = 14.69, Cl (8.60, 20.79), pc.001. 30-day average of time spent in REM sleep (per day) of femal30-dayicipants (n = 22) significantly increased from TO to T1 , mean difference = 13.88, Cl (6.68, 21.09), pc.001. TO to T2, mean difference = 11.57, Cl (3.99, 19.15), p = .003. TO to T3, mean difference = 12.12, Cl (4.54, 19.70), p = .002. 30-day average of time spent in REM sleep (per day) of students (n = 16) significantly increased from TO to T1 , mean difference = 1 1 .89, Cl (6.60, 17.18), pc.001. TO to T2, mean difference = 11.93, Cl (6.52, 17.34), pc.001. TO to T3, mean difference = 10.86, Cl (5.32, 16.40), pc.001. 30-day average of time spent in REM sleep (per day) ofworki30-daylts (n = 28) significantly increased from TO to T1 , mean difference = 11 .63, Cl (7.69, 15.57), pc.001. TO to T2, mean difference = 10.43, Cl (6.39, 14.48), p<.001. TO to T3, mean difference = 9.40, Cl (5.35, 13.45), pc.001.

[0229] Fitbit: Light Sleep Stage Counts

[0230] There were no significant changes in Light Sleep Stage Counts of all participants (n = 44) across all timepoints. There were no significant changes in Light Sleep Stage Counts of male participants (n = 22) across all timepoints. There were no significant changes in Light Sleep Stage Counts of female participants (n = 22) across all timepoints. There were no significant changes in Light Sleep Stage Counts of students (n = 16) across all timepoints. There were no significant changes in Light Sleep Stage Counts of working adults (n = 28) across all timepoints.

[0231] Fitbit: Deep Sleep Stage Counts

[0232] There were no significant changes in Deep Sleep Stage Counts of all participants (n = 44) across all timepoints. There were no significant changes in Deep Sleep Stage Counts of male participants (n = 44) across all timepoints. There were no significant changes in Deep Sleep Stage Counts of female participants (n = 22) across all timepoints. There were no significant changes in Deep Sleep Stage Counts of students (n = 16) across all timepoints. There were no significant changes in Deep Sleep Stage Counts of working adults (n = 28) across all timepoints.

[0233] Fitbit: REM Sleep Stage Counts

[0234] REM Sleep Stage Counts of all participants (n = 44) significantly decreased from TO to T3, mean difference = -.69, Cl (-1 .37, -.016), p = .045. REM Sleep Stage Counts of male participants (n = 22) significantly decreased from TO to T3, mean difference = -1 .19, Cl (-2.28, -.11), p = .032. There were no significant changes in REM Sleep Stage Counts of male participants (n = 22) across all timepoints. REM Sleep Stage Counts of students (n = 16) significantly decreased from TO to T3, mean difference = -1 .59, Cl (-3.14, -.03), p = .046. There were no significant differences in REM Sleep Stage Counts of working adults (n = 28).

[0235] Fitbit: Heart Rate Variability, Daily RMSSD

[0236] There were no significant changes in Heart Rate Variability, Daily RMSSD of all participants (n = 44) across all timepoints. There were no significant changes in Heart Rate Variability, Daily RMSSD of male participants (n = 22) across all timepoints. There were no significant changes in Heart Rate Variability, Daily RMSSD of female participants (n = 22) across all timepoints. There were no significant changes in Heart Rate Variability, Daily RMSSD of students (n = 16) across all timepoints. There were no significant changes in Heart Rate Variability, Daily RMSSD of working adults (n = 28) across all timepoints.

[0237] Fitbit: Heart Rate Variability, Deep RMSSD

[0238] Heart Rate Variability, Deep RMSSD of all participants (n = 44) significantly increased from TO to T3, mean difference = 3.45, Cl (.15, 6.75), p = .040. Heart Rate Variability, Deep RMSSD of male participants (n = 22) significantly increased from TO to T3, mean difference = 3.45, Cl (.15, 6.75), p = .040. Heart Rate Variability, Deep RMSSD of female participants (n = 22) significantly increased from TO to T3, mean difference = 3.45, Cl (.15, 6.75), p = .040. There were no significant differences in Heart Rate Variability, Deep RMSSD of students (n = 16) across all timepoints. Heart Rate Variability, Deep RMSSDof working adults (n = 28) significantly increased from TO to T3, mean difference = 4.75, Cl (1 .06, 8.44), p = .012.

[0239] Table : Summary of the Fitbit assessments

[0240] In the present study objective indicators of sleep were also observed to have significant improvements after the intake of Tongkat Ali. Total minutes of REM sleep measured from the Fitbit wearables indicated improvements after 6 weeks of Tongkat Ali intake for all participants. For working adults, the effects seemed to be more pronounced with a significant increase in total minutes of REM sleep after 3 weeks of Tongkat Ali intake. Improvements in 30-day-average of light, deep and REM sleepwas also observed at all timepoints for all participants, regardless of gender and working status.

[0241] A regression analysis performed on objective measurements of sleep (Fitbit) and participants' mood profile found that an increase in total minutes of deep sleep predicted a decrease in Depression- Dejection mood (POMS-2) 3 weeks after the intake of Tongkat Ali. Simple linear regression analysis was conducted to evaluate the extent to which participants’ total minutes of deep sleep could predict their POMS-2 Depression-dejection scores at T1 .

[0242] A significant regression was found, F(1 , 43) = 4.328, p = .043. The R2was .09, indicating that participants’ total minutes of deep sleep explained approximately 9% of the variance in their POMS-2 Depression-dejection scores. The regression equation was: POMS-2 Depression-dejection = 59.717 - 0.172 (Total minutes of deep sleep). That is, for each one-unit increase in a participant’s total minutes of deep sleep, the predicted POMS-2 Depression-dejection score decreased by 0.172. The confidence interval (Cl) indicated 95% certainty of the slope being able to predict POMS-2 Depression-dejection is between -0.339 and -0.005.

[0243] A significant regression was found in working adults, F(1 , 26) = 4.013, p = 0.004. The R2was 0.12, indicating that participants’ total minutes of deep sleep explained approximately 12% of the variance in their POMS-2 Depression-dejection scores. The regression equation was: POMS-2 Depressiondejection = 59.015 - 0.165 (Total minutes of deep sleep). That is, for each one-unit increase in a participant’s total minutes of deep sleep, the predicted POMS-2 Depression-dejection score decreased by 0.165. The Cl again indicated 95% certainty of the slope being able to predict POMS-2 Depressiondejection is between -0.366 and 0.056.

[0244] The intake of Tongkat Ali also seemed to have influenced the subjective and objective improvements in participants’ cognitive profile. Subjective measurements such as POMS-2 Confusion- Bewilderment and WHO-QoL Thinking, Learning, Memory and Concentration were observed to improve significantly across all timepoints.

[0245] Objective measurements of participants’ cognitive flexibility and working memory were observed to improve as well. More specifically, Stroop Task overall mean response time (all, congruent and incongruent tasks) significantly decreased across all timepoints regardless of gender and working status. This indicated an objective improvement in cognitive flexibility. Digitspan Backward Maximal and New measure also improved significantly after 6-weeks of Tongkat Ali intake, indicating an objective improvement in working memory.

[0246] Subjective improvements in participants’ quality of life across physical, psychological and social relationships domains were also observed after the intake of Tongkat Ali. Participants reported an improvement in Pain and Discomfort, Energy and Fatigue, Sleep and Rest, Positive Feelings, Self- Esteem, Body Image and Appearance, Negative Feelings, Sexual Activity quality of life (WHO-QoL).

[0247] It should be further appreciated by the person skilled in the art that variations and combinations of features described above, not being alternatives or substitutes, may be combined to form yet further embodiments falling within the intended scope of the invention.

[0248] As would be understood by a person skilled in the art, each embodiment, may be used in combination with other embodiment or several embodiments.

Claims

ClaimsClaim 1. A method of treating an individual diagnosed with insomnia or improving the mood, quality of life, or working memory of an individual in need comprising administering to the individual diagnosed with insomnia or provided to the individual in need a root extract of a Eurycoma longifolia plant.Claim 2. The method according to claim 1 , wherein the root extract of a Eurycoma longifolia plant comprises about 1.5% Eurycomanone, about 22% protein, about 30% polysaccharides and about 35% glycosaponin.Claim 3. The method according to claim 1 , wherein the root extract of a Eurycoma longifolia plant is formulated for oral administration or consumption.Claim 4. The method according to claim 1 , wherein the root extract of a Eurycoma longifolia plant is formulated as a capsule for oral administration.Claim 5. The method according to claim 1 , wherein the root extract of a Eurycoma longifolia plant is formulated as a liquid for oral administration.Claim 6. The method according to claim 1 , wherein the root extract of a Eurycoma longifolia plant is formulated as a supplement for oral consumption.Claim 7. The method according to claim 1 , wherein the root extract of a Eurycoma longifolia plant is formulated as food preparation for oral consumption.Claim 8. The method according to claim 1 , wherein the root extract of a Eurycoma longifolia plant is formulated as beverages for oral consumption.Claim 9. The method according to claim 1 , wherein the root extract of a Eurycoma longifolia plant is formulated as a single dose comprising 200mg.Claim 10. The method according to claim 1 , wherein 200mg of the root extract of a Eurycoma longifolia plant is administered daily.Claim 11 . A root extract of a Eurycoma longifolia plant for use in the treatment of insomnia.Claim 12. The root extract for use according to claim 11 , wherein the root extract comprises about 1.5% Eurycomanone, about 22% protein, about 30% polysaccharides and about 35% glycosaponin.Claim 13. The root extract for use according to claim 11 or 12, wherein the root extract of a Eurycoma longifolia plant is formulated for oral administration.Claim 14. The root extract for use according to any one of claim 11 to 13, wherein the root extract of aEurycoma longifolia plant is formulated for oral administration as a capsule or as a liquid.Claim 15. The root extract for use according to any one of claim 1 1 to 14, wherein the root extract is formulated as a dose comprising 200mg.Claim 16. The root extract for use according to any one of claim 1 1 to 15, wherein the root extract is formulated as a daily dose.Claim 17. Use of a root extract of a Eurycoma longifolia plant in the manufacture of a medicament for use in the treatment of insomnia.Claim 18. Use according to claim 17, wherein the root extract comprises about 1 .5% Eurycomanone, about 22% protein, about 30% polysaccharides and about 35% glycosaponin.Claim 19. Use according to claim 17 or 18, wherein the root extract of a Eurycoma longifolia plant is formulated for oral administration.Claim 20. Use according to any one of claim 17 to 19, wherein the root extract of a Eurycoma longifolia plant is formulated for oral administration as a capsule or as a liquid.Claim 21. Use according to any one of claim 17 to 20, wherein the root extract is formulated as a dose comprising 200mg.Claim 22. Use according to any one of claim 17 to 21 , wherein the root extract is formulated as a daily dose.

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