Use and method of orally active peptides for modulating sleep

Abstract

The present invention relates to the field of biotechnology and neuroscience. More specifically, the present invention discloses a new use of the Seq ID No 1 peptide (Asp- Ile-Ile-Ala-Asp-Asp-Glu-Pro-Leu-Thr, or DIIADDEPLT, also known as Pep19) for the preparation of a supplement or a medicament for sleep modulation. Also disclosed is a method for modulating sleep by administering, preferably orally, said peptide or a composition comprising said peptide.

Description

Technical Field

[0001] This invention relates to the fields of biotechnology and neuroscience. More specifically, this invention discloses a novel use of the peptide of Seq ID No1 (Asp-Ile-Ile-Ala-Asp-Asp-Glu-Pro-Leu-Thr, or DIIADEPLT, also known as Pep19) in the preparation of supplements or medicines for sleep regulation. A method for regulating sleep by preferably oral administration of the peptide or a composition containing the peptide is also disclosed. Background Technology

[0002] Insufficient sleep and poor sleep quality are widespread. In short, insufficient sleep and poor sleep quality impair overall health and manifest as a range of functional impairments, from depressed mood and cognitive impairment to metabolic disorders. These functional impairments translate into discomfort, loss of quality of life and / or health, and economic costs for affected individuals.

[0003] Strategies for addressing sleep deprivation and poor sleep quality include reconfiguring the sleep environment, behavioral training, exercise, and reducing caffeine intake. These methods also include the use of sleep aids, which can include prescription drugs and a range 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 prior art to discover additional strategies that can help eliminate sleep deprivation and improve sleep quality. This invention provides alternatives to these problems.

[0004] Peptide 19, or Pep19™, is a synthetic peptide developed by the applicant of this patent application and is the technical solution disclosed in International Patent Application PCT / BR2013 / 000251, WO 2014 00857, which has been granted in several countries. This peptide can be classified as one of the intracellular peptides (InPeps), which are produced by the proteasome hydrolysis of intracellular proteins and have cellular functions different from hormonal peptides and neuropeptides. Intracellular peptides are generally abundant, and their concentrations increase and decrease depending on circumstances: diet-induced obesity, induced non-alcoholic fatty liver disease, cell cycle, hypoxia, etc. However, the lack of mention of Pep19's potential use in alleviating sleep deprivation and / or improving sleep quality in the aforementioned literature is unexpected even to the inventors.

[0005] A search of prior art in scientific and patent literature revealed no documents that presuppose or suggest the teachings of this invention.

[0006] Based on the literature searched, no literature was found that anticipated or suggested the teachings of the present invention. Summary of the Invention

[0007] This invention provides a novel use for 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. This invention also provides a method for regulating sleep by preferably oral administration of the peptide or a composition containing the peptide.

[0008] Therefore, one of the objects of this invention is to provide a novel use of the peptide of SEQ ID NO:1 for the preparation of supplements or medicines for sleep regulation.

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

[0010] These and other objects of the present invention will be readily understood by those skilled in the art and will be described in detail below. Attached Figure Description

[0011] Figure 1 A graph showing the effect of Pep19 administration on sleep quality over time is shown. A) shows the change in total score on day 60, expressed as change (%) relative to baseline (mean ± SEM). Pep19 vs. placebo; p < 0.05). Figure B) shows the number of individuals with a PSQI score higher than 5 (out of 0) 60 days before and after starting Pep19 administration.

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

[0013] Figure 3The graphs show the effects of Pep19 administration over time on sleep quality in relation to the following parameters: 1) subjective sleep quality; 2) sleep latency; 3) sleep duration; 4) usual sleep efficiency; 5) sleep disturbances; 6) use of sleep medications; and 7) daytime dysfunction. In A), initial score values ​​of 0, 1, 2, and 3 are shown for each subjective and objective parameter analyzed in each administration regimen: placebo or 0 mg; 2 mg, and 5 mg (represented by circles from left to right, respectively). In B), score values ​​of 0, 1, 2, and 3 are shown after 60 days for the same parameter. In this analysis, a score of 0 (zero) categorized sleep quality as good, and a score of 3 (three) categorized sleep quality as poor. Detailed Implementation

[0014] This invention provides a novel use for the SEQ ID NO:1 peptide or Asp-Ile-Ile-Ala-Asp-Asp-Glu-Pro-Leu-Thr (DIIADDEPLT), which addresses the technical problem of sleep difficulties in mammals. This invention also provides a method for regulating sleep by preferably oral administration of the peptide or a composition containing the peptide.

[0015] The inventors conducted preliminary experiments and, based on findings from individual self-reports, demonstrated that administration of Pep19 to obese individuals resulted in improved sleep. This result encouraged the inventors to test this unexpected effect in greater detail.

[0016] In this invention, it was unexpectedly demonstrated that oral administration of peptide 19 or Pep19 significantly improved sleep quality in humans. In a human clinical trial, participants receiving a blinded placebo were randomized to take 2 mg or 5 mg of Pep19 as a single capsule at bedtime for 60 days, followed by sleep quality assessment using the Pittsburgh Quality Sleep Index (PSQI) and quality of life assessment using the SF-12 questionnaire (12-Item Short-Form Health Survey). As a result, there were no adverse events, no health-related changes in quality of life, and no biochemical toxicity. Statistically significant improvements in sleep quality were reported in 35% (SEM=10%) and 25% (SEM=16%) of the 2 mg and 5 mg groups, respectively. Specifically, the PSQI score of individuals with a score higher than 5 (the threshold between good and poor sleep quality; a score higher than 5 is considered poor sleep) decreased from 6 to 3 in the 2 mg group and from 8 to 3 in the 5 mg group. The results were quite unexpected, indicating that Pep19 is well-tolerated in humans and significantly improves sleep quality, providing a new approach for those suffering from sleep disorders.

[0017] Example 1 - Human Clinical Trial The Advarra Institutional Review Board approved the study protocol for the impact of Pep19 intake on quality of life. Individuals were recruited from primary care clinics in Fort Lauderdale, Florida, and their medical conditions, ranging from systemic symptoms (fatigue) to organ-specific symptoms (urinary frequency), were assessed, as is common in systemic medical history reviews.

[0018] Recruitment criteria typically include healthy adults aged 40 to 60, i.e., adults residing in the community, who can attend clinics for eligibility screening and follow-up appointments, and whose body mass index (BMI) is between 30 and 35 kg / m². 2 Exclusion criteria include pregnancy and use of anti-obesity supplements or medications, such as smegglutinin.

[0019] Informed consent was obtained from 24 individuals based on the form submitted to Advara IRB Protocol Pro00075623. They were then randomly assigned in a 60-day double-blind model to receive either 5 mg or 2 mg of Pep19 daily in a 1:1:1 blinded manner, or a placebo. Pep19 was formulated as a dietary supplement in vegetable capsules containing microcrystalline cellulose excipients, the proportions of which are shown in Table 1.

[0020] Table 1 - Formulations used in this phase of clinical trials (%wt) In other assessments conducted as parallel research subjects (not reported in this patent application), health-related quality of life was measured using the 12-item Health Survey Short Form (SF-12). Anthropometric measurements and vital signs were performed by staff at the clinic. Body composition was measured using dual-energy X-ray absorptiometry (DEXA). Blood biochemistry included fasting blood glucose, insulin, HgBA1c, ALT, AST, C-reactive protein, and triglycerides.

[0021] The statistician remained blinded during the initial analysis of the raw data, receiving a number only after the sleep quality analysis (and all other data not reported in this patent application, such as blood pressure, weight, and waist circumference) was completed. Data were collected and analyzed using GraphPad Prism (GraphPad software, www.graphpad.com) via univariate or two-way ANOVA analysis with Tukey's multiple comparison test. Significance was set at p < 0.05.

[0022] Table 2 shows the demographic data and baseline measurements of the experimental group before the administration of Pep19.

[0023] Table 2 - Demographic data and baseline measurements (±SD). At this initial point, there were no statistically significant differences between the two groups. (body fat / height) 2 ) The improvement in sleep after Pep19 administration can be seen in Figure 1 The results are shown in graph form. A) shows the change in sleep quality relative to baseline. B) shows the number of individuals with a score higher than 5 for each administration regimen: a) placebo or 0 mg; b) 2 mg; and c) 5 mg (x-axis, time in days).

[0024] The sleep improvement results from administering Pep19 can also be seen in... Figure 2 The results, presented in chart form, show the percentage of individuals with PSQI scores of 0 to 5 (good sleep quality) at baseline (T0) and after 60 days (T60) for each administration regimen: placebo or 0 mg; 2 mg; and 5 mg, represented by gray bars, and those with scores higher than 5 (poor sleep quality) at baseline (T0) and after 60 days (T60) by black bars. The results were quite unexpected and highlight the fact that Pep19 significantly improves sleep quality, providing a new approach for those suffering from sleep disorders.

[0025] exist Figure 3 The results of improved sleep through Pep19 administration were fully analyzed. A) shows the initial score values ​​of 0, 1, 2, and 3 for each subjective and objective parameter analyzed in each administration regimen: placebo or 0 mg; 2 mg, and 5 mg (represented by circles from left to right). B) shows the scores of 0, 1, 2, and 3 after 60 days for the same parameter. In this analysis, a score of 0 (zero) categorized sleep quality as good, and a score of 3 (three) categorized sleep quality as poor.

[0026] These analyses show that Pep19 improves sleep parameters, particularly insomnia (without causing daytime sleepiness or nighttime disorientation), overcoming the difficulties faced by other sleep-enhancing drugs in terms of the absence of side effects.

[0027] Apart from Figures 1 to 3In addition to the results shown, a significant reduction in visceral fat was observed in the 5 mg Pep19 group (83 ± 5% compared to baseline at 60 days, compared to 102 ± 5% in the placebo group at 60 days). No significant difference in visceral fat reduction was observed in the 2 mg Pep19 group. However, the results regarding sleep improvement were more pronounced in the 2 mg Pep19 group, indicating no direct relationship between visceral fat reduction and sleep improvement.

[0028] The potential mechanisms by which Pep19 improves sleep quality are not yet clear. It does not appear to reflect a direct effect on the brain, which regulates sleep quality, as previous data from animals showed that Pep19 had no effect on the brain. Therefore, the inventors speculate that these may be indirect effects. While the experiments conducted to date have significant limitations, their conclusions are based on statistically significant results, although they have been conducted for a relatively short period and do not elucidate the effects of Pep19 after two months. However, these results confirm previous reports from randomized, unconnected Pep19 users who unexpectedly showed improved sleep after administration of Pep19 or compositions containing Pep19. Therefore, the findings offer a new option for people with poor sleep quality. The method is relatively simple—one capsule before bedtime—and there have been no apparent adverse events, meaning that absorption faces few major obstacles.

[0029] Those skilled in the art will understand the knowledge presented herein and will be able to reproduce the invention in the illustrated embodiments and other variations and alternatives covered by the scope of the appended claims.

Claims

1. Use of a peptide of SEQ ID NO 1, characterized in that, It is used to prepare supplements or drugs for sleep regulation.

2. Use according to claim 1, characterized in that, It is used to prepare oral supplements or drugs for sleep regulation.

3. A method for modulating sleep in a mammal, characterized in that, It includes a therapeutically effective amount of the peptide of SEQ ID NO 1.

4. The method of claim 3, wherein, This includes administering 2 to 5 mg to human individuals daily.

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