Use of orally active peptide and method for modulating sleep

Abstract

The present invention pertains to the fields of biotechnology and neuroscience. More specifically, the invention discloses a novel use of the peptide SEQ ID NO:1 (Asp-Ile-Ile-Ala-Asp-Asp-Glu-Pro-Leu-Thr, or DIIADDEPLT, also known as Pep19) for the preparation of a supplement or drug for sleep modulation. Also disclosed is a method for modulating sleep preferably through oral administration of said peptide or a composition comprising same.

Description

Descriptive Report of Invention Patent USE OF ORALLY ACTIVE PEPTIDE AND METHOD FOR MODULATING SLEEP Field of the Invention

[0001] The present invention is in the field of Biotechnology and Neuroscience. More specifically, the invention discloses a novel use of the peptide of Seq ID No. 1, (Asp-Ile-Ile-Ala-Asp-Asp-Glu-Pro-Leu-Thr, or DIIADDEPLT, also known as Pep19, for the preparation of a supplement or medicament for the modulation of sleep. A method for modulating sleep through the preferentially oral administration of said peptide or a composition comprising it is also disclosed. Background of the Invention

[0002] Sleep deprivation and poor sleep quality are epidemic. Together, sleep deprivation and poor sleep quality impair overall well-being and manifest in a range of dysfunctions ranging from moodiness, impaired cognition, and disrupted metabolism. These dysfunctions translate into discomfort, loss of quality of life and / or health, and economic costs for the affected individual.

[0003] Strategies for managing sleep deprivation and poor sleep quality include reconfiguring the sleep environment, behavioral training, exercise, reducing caffeine intake, and more. Such approaches also include the administration of sleeping pills, which can include prescription medications and a variety of over-the-counter medications, such as diphenhydramine, and supplements, such as melatonin, chamomile, and valerian. Each strategy has its limitations, including limited effectiveness. Therefore, there remains a significant need in the art to identify additional strategies that can help eliminate sleep deprivation and improve sleep quality. The present invention provides an alternative to these approaches. problems.

[0004] Peptide 19 or Pep19™ is a synthetic peptide developed by the applicant of this patent application, and is the subject of international patent application PCT / BR2013 / 000251, published as WO 201400857 and already granted in several countries. This peptide can be classified as one of the intracellular peptides (InPeps), which are generated from the proteasomal proteolysis of intracellular proteins and have cellular functions that differ from those of hormonal peptides and neuropeptides. InPeps are often abundant, and their concentrations increase and decrease depending on the circumstances: diet-induced obesity, inducible non-alcoholic fatty liver disease, cell cycle, hypoxia, and so on. However, nothing is mentioned in the literature about the potential use of Pep19 to help eliminate sleep deprivation and / or to improve sleep quality, making such a use surprising even to the present inventors.

[0005] In the search for the state of the art in scientific and patent literature, no documents were found anticipating or suggesting the teachings of the present invention.

[0006] From what can be inferred from the literature researched, no documents were found anticipating or suggesting the teachings of the present invention. Summary of the Invention

[0007] The present invention provides a novel use of the peptide of SEQ ID NO: 1, or Asp-Ile-Ile-Ala-Asp-Asp-Glu-Pro-Leu-Thr (DIIADDEPLT), which solves the technical problem of sleep difficulties in mammals. The invention also provides a method for modulating sleep through the preferably oral administration of said peptide or a composition comprising it.

[0008] It is, therefore, one of the objects of the invention to provide a new use of the peptide of SEQ ID NO: 1, for the preparation of a supplement or medicine for sleep modulation.

[0009] It is another object of the invention to provide a method for modulating sleep in mammals, comprising administering a therapeutically effective amount of the peptide of SEQ ID NO: 1.

[0010] These and other objects of the invention will be immediately appreciated by those skilled in the art and will be described in detail below. Brief Description of the Figure

[0011] Figure 1 presents graphs demonstrating the effect of Pep19 administration on sleep quality over time. In A) the change in total score at day 60 is shown, in terms of change (%) relative to baseline (mean ± SEM; *Pep19 vs placebo; p<0.05). In B) the number of individuals with a PSQI score above 5 out of 0 before 60 days after starting Pep19 administration is shown.

[0012] Figure 2 presents graphs showing the effect of Pep19 administration on sleep quality over time, where the percentage of individuals with a PSQI score of 0 to 5 (good sleep quality) is represented in gray and a score above 5 (poor sleep quality) is represented by the black bar at the initial time (T0) and after 60 days (T60) in each administration regimen: placebo or 0 mg; 2 mg and 5 mg.

[0013] Figure 3 presents graphs demonstrating the effect of Pep19 administration on sleep quality over time in relation to the following parameters: 1) Subjective sleep quality; 2) Sleep latency; 3) Sleep duration; 4) Habitual sleep efficiency; 5) Sleep disturbances; 6) Use of sleeping medication; and 7) Daytime dysfunction. In A) the initial scores of 0, 1, 2, and 3 are shown for each of the subjective and objective parameters analyzed in each administration regimen: placebo or 0 mg, 2 mg, and 5 mg (represented by a circle from left to right, respectively). In B) the scores of 0, 1, 2, and 3 after 60 days are shown for the same parameters. In these analyses, a score of 0 (zero) classifies sleep quality as good, and a score of 3 (three) classifies sleep quality as poor. Detailed Description of the Invention

[0014] The present invention provides a novel use of the peptide of SEQ ID NO: 1, or Asp-Ile-Ile-Ala-Asp-Asp-Glu-Pro-Leu-Thr (DIIADDEPLT), which solves the technical problem of sleep difficulties in mammals. The invention also provides a method for modulating sleep through the preferably oral administration of said peptide or a composition comprising it.

[0015] The present inventors conducted pilot experiments and demonstrated that administering Pep19 to obese individuals provided, according to the individuals' own self-reported improvement in sleep. This result encouraged the inventors to further test this surprising effect.

[0016] The present invention, surprisingly, demonstrates that oral administration of peptide 19 or Pep19 in humans significantly improves sleep quality. In a human clinical trial, randomized to receive placebo, administration of 2 mg or 5 mg of Pep19 as a single capsule at bedtime for 60 days was followed by sleep quality assessment using the Pittsburgh Quality Sleep Index (PSQI) and quality of life assessment using the SF-12 (12-Item Short-Form Health Survey). Results revealed no adverse events, no changes in health-related quality of life, and no biochemical toxicity. Statistically significant improvements of 35% (SEM=10%) and 25% (SEM=16%) in sleep quality were reported for the 2 mg and 5 mg groups, respectively.Specifically, the number of individuals with a PSQI score above 5 (the borderline between good and poor sleep quality; a score above 5 is considered poor sleep) dropped from 6 to 3 in the 2 mg group and from 8 to 3 in the 5 mg group. These striking results suggest that Pep19 is well tolerated in humans and significantly improves sleep quality, offering a new approach for those with sleep disorders.

[0017] Example 1 – Human clinical trial

[0018] The study protocol was approved by the Advarra Institutional Review Board for the Effect of Pep19 Consumption on Quality of Life study. Subjects were recruited from a primary care clinic in Fort Lauderdale, Florida, and assessed for medical conditions, as is typical in a system-wide medical history review, ranging from constitutional (fatigue) to organ-specific (urinary frequency).

[0019] Inclusion criteria were generally healthy adults between 40 and 60 years of age, i.e., adults living in the community and able to come to the clinic for qualification and follow-up appointments, and a body mass index (BMI) between 30 and 35 kg / m 2 . Exclusion criteria included pregnancy and use of anti-obesity supplements or medications, e.g., semaglutide.

[0020] Twenty-four individuals provided informed consent according to the form submitted to Advarra IRB protocol Pro00075623. They were then randomized to receive a blinded daily dose of 5 mg or 2 mg of Pep19 or placebo in a 1:1:1 ratio in a 60-day double-blind design. Pep19 was formulated as a dietary supplement in vegetarian capsules with microcrystalline cellulose excipient in the proportions shown in Table 1.

[0021] Table 1 – Formulations used in this phase of clinical trials (% wt) Placebo 2mg 5mg Pep19 0 1 3 Excipient 74 73 72 Capsule 26 26 26

[0022] Among other assessments that were the subject of a parallel study (not reported in this patent application), health-related quality of life was measured with the 12-Item Short Form Health Survey (SF-12). Anthropometric measurements and vital signs were taken by clinic staff. Body composition was measured with dual-energy X-ray absorptiometry. (DEXA). Blood chemistry included fasting glucose, insulin, HgBA1c, ALT, AST, C-reactive protein, and triglycerides.

[0023] The statistical analyst remained blinded during the initial analyses of the raw data and received the code only after completing the sleep quality analysis (and all other data not reported in this patent application, such as blood pressure, weight, and waist circumference). Data were collected and analyzed using one-way or two-way ANOVA with Tukey's multiple comparisons test using GraphPad Prism (GraphPad Software, www.graphpad.com). Significance was defined as p < 0.05.

[0024] Table 2 shows the demographic data and baseline measurements of the test groups, i.e., before Pep19 administration.

[0025] Table 2 – Demographic data and measurements at baseline (±SD). At this initial point there was no statistical difference between the groups. Placebo 2mg (n=7) 5mg (n=8) (n=8) Age (years) 55±4 53±5 52±6 Women 5 5 2 Men 3 2 6 Weight (kg) 94±12 91±10 100±6 Height (cm) 169±10 165±7 178±6 BMI 33±1 33±1 32±1 Fat Index* (g / cm2 ) 1.46±0.22 1.51±0.20 1.20±0.22 Sleep Quality (PSQI) 7±3 9±3 8±2 * (fat mass / height 2 )

[0026] The results of sleep improvement through Pep19 administration can be seen graphically in Figure 1. In A) the values ​​of change in sleep quality in relation to baseline are shown. In B) the results of the number of individuals with a score above 5 in each administration regimen are shown: a) placebo or 0 mg; b) 2 mg and c) 5 mg (and on the x-axis time in days).

[0027] The sleep improvement results from Pep19 administration can also be seen in the graph in Figure 2, which represents the percentage of individuals with a PSQI score of 0 to 5 (good sleep quality), represented in gray, and a score above 5 (poor sleep quality), represented by the black bar, at baseline (T0) and after 60 days (T60) in each administration regimen: placebo or 0 mg, 2 mg, and 5 mg. The results are quite surprising and point to the fact that Pep19 significantly improves sleep quality, offering a new approach for those with sleep disorders.

[0028] The results of sleep improvement through Pep19 administration were analyzed in detail in Figure 3. In A) the initial score values ​​0, 1, 2, and 3 are shown for each of the subjective and objective parameters analyzed in each administration regimen (placebo or 0 mg, 2 mg, and 5 mg) (represented by a circle from left to right, respectively). In B) the score values ​​0, 1, 2, and 3 after 60 days are shown for the same parameters. In these analyses, a score of 0 (zero) classifies sleep quality as good, and a score of 3 (three) classifies sleep quality as poor.

[0029] These analyses demonstrated that Pep19 improves sleep parameters and particularly insomnia (without causing drowsiness during the day or disorientation at night), overcoming the difficulty of other sleep-improving medications in not presenting side effects.

[0030] In addition to the results shown in Figures 1 to 3, a significant reduction in visceral fat was also noted in the group administered 5mg of Pep19 (83±5% compared to baseline after 60 days, versus 102±5% compared to baseline after 60 days in the placebo group). No significant difference in visceral fat reduction was observed in the group administered 2mg of Pep19. However, the sleep improvement results were more pronounced in the group administered 2mg of Pep19, which suggests that there is no direct relationship between visceral fat reduction and improved sleep.

[0031] The mechanism underlying the improved sleep quality with Pep19 administration is unknown. It does not appear to reflect a direct effect on the brain, where sleep quality is regulated, because previous animal data show that Pep19 has no effect on the brain. Consequently, the inventors speculate that it may have indirect effects. Although the experiments conducted thus far have significant limitations, their conclusions are based on statistically significant results, even though they were conducted (so far) for a relatively short period of time, and do not clarify the effects of Pep19 after two months. However, the present results corroborate previous reports from random, unconnected users of Pep19 that indicated surprisingly improved sleep after administration of Pep19 or compositions containing it. The results, therefore, provide a new alternative for people suffering from poor sleep.The relative simplicity of the method—administering one capsule at bedtime—and the apparent lack of adverse events mean that absorption faces few significant barriers.

[0032] Those skilled in the art will appreciate the knowledge presented herein and will be able to reproduce the invention in the presented embodiments and in other variants and alternatives, covered by the scope of the following claims.

Claims

1 / 1 Claims 1. Use of the peptide of SEQ ID NO 1 characterized in that it is for the preparation of a supplement or medicament for sleep modulation.

2. Use according to claim 1 characterized in that it is for the preparation of an oral supplement or medicament for sleep modulation.

3. Method for sleep modulation in mammals characterized in that it comprises the administration of a therapeutically effective amount of the peptide of SEQ ID NO 1.

4. Method according to claim 1 characterized in that it comprises the administration of 2 to 5 mg per day to a human individual.

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