Probiotic composition for the conditioning of sleep disorders

Abstract

The present application belongs to the technical field of probiotics, and discloses a regulating effect of a probiotic composition on sleep disorders. The probiotic composition contains Clostridium butyricum JSIM-MCB20040312 with a preservation number of CGMCC No. 1647 and Bacillus coagulans JSSW-LA-07 with a preservation number of CGMCC No. 2602. The strains have high safety, are not prone to drug resistance, and have a synergistic effect. The probiotic composition can significantly improve the difficulty of falling asleep, effectively improve the sleep quality, and effectively improve the daytime emotional and drowsiness impairment caused by sleep deprivation.

Description

Technical Field

[0001] This invention belongs to the field of probiotic technology, specifically a probiotic composition and its application. Background Technology

[0002] Sleep is an indispensable and normal physiological process for the human body, playing a crucial role in maintaining and improving multiple dimensions of physical and mental health. Insufficient sleep or interrupted sleep can easily lead to serious emotional disturbances and cognitive impairment. The duration and quality of sleep are influenced by a combination of cultural, social, psychological, behavioral, pathological, physiological, and environmental factors. A consensus statement jointly issued by the American Academy of Sleep Medicine and the Sleep Research Society clearly recommends that adults get at least 7 hours of sleep each night. However, with social development, increasingly diverse entertainment options, the normalization of overtime work, and increasing psychological and social pressures, people's sleep time may be decreasing and sleep quality may be significantly declining.

[0003] Currently, public health departments are paying increasing attention to sleep problems. In 2022, the American Heart Association introduced the concept of the "Eight Elements of Life," formally incorporating sleep health into the key points of cardiovascular disease prevention, compared to the "Seven Simple Things to Live" in 2010. Furthermore, numerous prospective and cross-sectional studies have confirmed that sleep plays a crucial role in the occurrence and development of diseases such as hypertension, diabetes, dyslipidemia, obesity, and metabolic syndrome; both abnormal sleep quality and abnormal sleep duration may be closely related to these diseases.

[0004] Currently, sleep assessment methods are diverse, broadly categorized into subjective and objective assessments. Subjective sleep assessments often employ scales, with the Pittsburgh Sleep Quality Index (PSQI), Insomnia Severity Index (ISI), and Epworth Sleepiness Scale (ESS) being widely used in clinical practice. Objective sleep assessments primarily utilize polysomnography (PSG) and the Multiple Sleep Latency Test (MSLT). The Athens Insomnia Scale is a commonly used tool for patients and clinicians to subjectively assess sleep symptoms. By comprehensively analyzing patient symptoms, relevant scale assessment results, and polysomnography data, clinicians can often accurately diagnose and classify various types of sleep disorders.

[0005] Insomnia is a sleep disorder characterized by frequent and persistent difficulty initiating and / or maintaining sleep, often accompanied by excessive worry about sleep, low sleep satisfaction, or impaired daytime functioning. The prevalence of insomnia in the general population of China is 15%-22%, and approximately 40% of insomnia patients develop chronic, persistent insomnia, resulting in high direct and indirect annual medical costs. In addition to the heavy social burden, long-term insomnia seriously affects the physical and mental health of patients, increasing the risk of various health problems, potentially exacerbating or increasing the incidence of mental disorders such as depression, and even increasing the risk of suicide. Therefore, exploring and proposing effective methods to improve insomnia is of crucial practical significance.

[0006] *Clostridium butyricum* (C. butyricum) is a Gram-positive anaerobic spore-forming bacillus belonging to the phylum Firmicutes, class Clostridia, order Clostridales, family Clostridiaceae, and genus *Clostridium*. This strain is heat-, pressure-, and bile-tolerant. It produces spores, possesses flagella, and is motile, making it a beneficial symbiotic bacterium for the intestines of both humans and animals. Its metabolic products include digestive enzymes, short-chain fatty acids (SCFAs), and amino acids, which effectively maintain the stability of the intestinal microecology.

[0007] Bacillus coagulans (B. coagulans) is a Gram-positive, anaerobic spore-forming bacillus. It possesses both spore-forming and lactic acid-producing properties, exhibiting not only the probiotic characteristics of Bifidobacteria and lactic acid bacteria, but also extremely strong heat, acid, and bile salt resistance. This gives Bacillus coagulans a unique advantage over other lactic acid bacteria in food processing, storage, and transportation. In 2016, my country included Bacillus coagulans in the "List of Edible Fungi," further verifying its safety. Bacillus coagulans can survive well in a temperature range of 35-50℃, and its optimal growth pH is 5.5-6.5. Its probiotic properties are mainly reflected in its promotion of the host's intestinal health: first, by regulating the balance of intestinal flora, inhibiting the growth of harmful bacteria, and maintaining the balance of intestinal microecology; second, by promoting the absorption and utilization of nutrients, and improving the host's utilization rate of nutrients; and third, by stimulating the development and functional improvement of the intestinal immune system, enhancing the host's immunity, and reducing the risk of intestinal diseases.

[0008] Bacillus coagulans creates an anaerobic environment for Clostridium butyricum through aerobic metabolism. Clostridium butyricum secretes glycosidases to degrade fiber and produce glucose, which provides energy substrates for Bacillus coagulans to produce lactic acid. The two interact to form a "metabolic complementary cycle." However, whether the combined use of Clostridium butyricum and Bacillus coagulans has an improving effect on sleep disorders has not yet been investigated. Summary of the Invention

[0009] This invention provides a probiotic composition that can improve sleep disorders. This probiotic composition significantly reduces insomnia rates and effectively improves sleep quality while enhancing overall health, exhibiting high safety and outstanding therapeutic effects.

[0010] In a first aspect, the present invention provides the use of a probiotic composition in the preparation of an agent for improving sleep disorders. The probiotic composition contains the following strains: *Clostridium butyricum* JSIM-MCB 20040312 (CGMCC No. 1647) and *Bacillus coagulans* JSSW-LA-07 (CGMCC No. 2602). *Clostridium butyricum* is also known as butyric acid bacteria, *Clostridium butyricum*, or butyric acid bacterium.

[0011] Furthermore, the probiotic composition contains Clostridium butyricum and Bacillus coagulans in a live bacteria ratio of 1:5-30 CFU / g.

[0012] Furthermore, the probiotic composition contains Clostridium butyricum and Bacillus coagulans in a live bacteria ratio of 1:20 CFU / g.

[0013] Furthermore, the probiotic composition contains Clostridium butyricum and Bacillus coagulans in a live bacteria ratio of 1:15 CFU / g.

[0014] Furthermore, the probiotic composition contains Clostridium butyricum and Bacillus coagulans in a live bacteria ratio of 1:10 CFU / g.

[0015] Furthermore, the concentration of Clostridium butyricum and Bacillus coagulans in the probiotic composition is 1×10⁻⁶. 5 Up to 1×10 11 CFU / g.

[0016] Furthermore, the sleep disorders include a range of conditions such as insomnia and daytime functional impairment due to insufficient sleep.

[0017] Furthermore, the aforementioned insomnia refers to a syndrome in which an individual, despite having suitable sleep conditions and environment, still experiences difficulty falling asleep, difficulty maintaining sleep, early awakening, or decreased sleep quality, and this condition persists and has a significant impact on daytime social functioning.

[0018] Furthermore, the preparation can be in the form of a drug, food, food additive, or other similar formulation.

[0019] The Bacillus coagulans of this invention has been deposited with the China General Microbiological Culture Collection Center (CGMCC) at the Institute of Microbiology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing. The deposit name is Bacillus coagulans JSSW-LA-07, the accession number is CGMCC No. 2602, and the deposit date is July 28, 2008.

[0020] The *Clostridium butyricum*, also known as *Clostridium butyricum*, in this invention has been deposited with the China General Microbiological Culture Collection Center (CGMCC). The deposit address is No. 13, Zhongguancun North First Street, Haidian District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. The deposit name is *Clostridium butyricum* JSIM-MCB 20040312, the accession number is CGMCC No. 1647, and the deposit date is March 10, 2006.

[0021] Beneficial effects: The strains used in this invention are all probiotics, which are highly safe and unlikely to develop drug resistance. An interaction was found between the two, allowing them to work synergistically to improve sleep quality, specifically: (1) significantly improving sleep pattern satisfaction and reducing difficulty falling asleep; (2) significantly improving mental fatigue and weakness, and improving behavioral activity and responsiveness. Therefore, this probiotic composition provides a new strategy for improving sleep quality and can be used to prepare products for the prevention, relief, or treatment of insomnia, with broad application prospects. The probiotic preparation of this invention can significantly improve difficulty falling asleep, increase sleep quality satisfaction, and reduce daytime sleepiness. Attached Figure Description

[0022] Figure 1 A graph showing the change in individual rank during the first month of follow-up for the ISI scale control group. Figure 2 Chart showing the change in individual rank during the first month of follow-up for the ISI scale treatment group. Figure 3 Chart showing the change in individual rank in the control group of the ISI scale during the third month of follow-up. Figure 4 Chart showing the change in individual rank in the treatment group at the third month of follow-up using the ISI scale. Figure 5 Graph showing the change in individual rankings of the ISI scale control group during the sixth month of follow-up. Figure 6 Chart showing the change in individual rank in the ISI scale treatment group at the sixth month of follow-up. Detailed Implementation

[0023] The specific embodiments of the present invention will be further described below. It should be noted that these descriptions are for the purpose of aiding understanding the present invention, but do not constitute a limitation thereof. Furthermore, the technical features involved in the embodiments described below can be combined with each other as long as they do not conflict with each other.

[0024] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods, and the experimental materials used in the following embodiments are all available through conventional commercial channels.

[0025] Example 1: Evaluation of Probiotic Efficacy In this embodiment of the invention, the probiotic strains used were all from Jiangsu Suwei Microbial Research Co., Ltd. Corn starch was a commercially available pharmaceutical excipient.

[0026] Preparation of probiotic formulations 1.1 Preparation of Clostridium butyricum preparations In this embodiment, the probiotics are live Clostridium butyricum JSIM-MCB20040312 (CGMCC No. 1647) and corn starch, wherein the concentration of Clostridium butyricum is 1×10⁻⁶. 8 CFU / g was used to make Clostridium butyricum capsules, each capsule containing 315mg of Clostridium butyricum JSIMMCB20040312 and 35mg of corn starch.

[0027] 1.2 Preparation of Bacillus coagulans preparation In this embodiment, the probiotics are live Bacillus coagulans JSSW-LA-07 (CGMCC No. 2602) and corn starch, wherein the concentration of Bacillus coagulans is 1×10⁻⁶. 8 CFU / g, formulated into Bacillus coagulans capsules, each capsule containing 315mg Bacillus coagulans JSSW-LA-07 and 35mg corn starch.

[0028] 1.3 Preparation of probiotic composition formulations The probiotic composition of this embodiment consists of the following components: live Bacillus coagulans JSSWLA07 (CGMCC No. 2602), live Clostridium butyricum JSIMMCB20040312 (CGMCC No. 1647), and corn starch; wherein the concentration of live Bacillus coagulans JSSWLA07 is 1×10⁻⁶. 8 The concentration of live bacteria of Clostridium butyricum JSIMMCB20040312 (CFU / g) is 1×10⁻⁶. 8CFU / g, *Clostridium butyricum* and *Bacillus coagulans* were mixed in a 1:10 ratio. The corn starch content in the probiotic composition was 10% of the total probiotic composition, i.e., 28.6 mg of *Clostridium butyricum* and 286.4 mg of *Bacillus coagulans* were mixed, and 35 mg of corn starch was added. The probiotic composition composed of the above components was formulated into oral capsules, each containing 350 mg of the probiotic composition.

[0029] 2. Research Subjects Sixty-eight patients with insomnia from Shanghai Fourth People's Hospital were selected. The drug administration study was conducted primarily through clinical follow-up and in a double-blind manner. All methods were performed in accordance with relevant guidelines and regulations.

[0030] 3. Western medicine diagnostic criteria: Diagnostic criteria for insomnia: Refer to the diagnostic criteria and classification principles for chronic insomnia disorder in the International Classification of Sleep Disorders (Third Edition). (1) Chief complaint (one or more of the following criteria are met): 1) Difficulty falling asleep; 2) Inability to maintain sleep for a long time; 3) Early awakening; 4) Inability to fall asleep at an appropriate time; 5) Inability to sleep alone; (2) Daytime symptoms (one or more of the following criteria are met): 1) General malaise or fatigue; 2) Impaired attention or memory loss; 3) Impaired social, household, occupational, or academic abilities; 4) Mood disturbance / irritability; 5) Daytime sleepiness; 6) Behavioral problems such as hyperactivity, aggression, or impulsiveness; 7) Decreased motivation, energy, or work initiative; 8) Prone to errors or accidents; 9) Concern or dissatisfaction with sleep. (3) Frequency of occurrence: Sleep disturbances and related daytime symptoms occur at least 3 times per week; Disease course: Sleep disturbances and related daytime symptoms have been present for at least 3 months; ① Diagnostic criteria for chronic insomnia: A diagnosis can be made if the criteria (1) to (6) are met.

[0031] ② Diagnostic criteria for short-term insomnia: Based on the diagnosis of chronic insomnia, the duration of the disease is <3 months.

[0032] 4. Inclusion and Exclusion Criteria 4.1 Inclusion Criteria 1. Meets the diagnostic criteria for chronic insomnia disorder according to the International Classification of Sleep Disorders (Third Edition); 2. Able to read, understand, follow research procedures, and independently complete a sleep diary; 3. Patients voluntarily participate in clinical trials and sign informed consent forms.

[0033] 4.2 Exclusion Criteria 1. Comorbid sleep disorders and mental disorders, such as sleep apnea syndrome, obsessive-compulsive disorder, schizophrenia, bipolar disorder, etc.; 2. Individuals experiencing a cold with fever or fever of unknown origin, with a body temperature ≥37.3℃; 3. Individuals with drug or alcohol dependence, addiction, or allergy to the drugs used in this study; 4. Do not take any hypnotics, antidepressants, central nervous system stimulants, or other traditional Chinese medicines or proprietary Chinese medicines that have sleep-improving effects, or other insomnia treatments, within one week before the induction period or within five half-lives of the drug. 5. Those currently taking other clinical trial drugs, or those whose washout period after stopping the trial is less than 2 weeks; 6. Accompanied by severe gastrointestinal diseases, such as gastrointestinal obstruction, gastric ulcers, and other gastrointestinal diseases that affect drug absorption; 7. Patients with concurrent liver or kidney dysfunction, severe neurological diseases, cardiovascular diseases, endocrine diseases such as diabetes and hyperthyroidism, autoimmune diseases, or other contraindications; 8. Women who are pregnant, breastfeeding, or may become pregnant during the research; 9. Individuals who, in the researchers' opinion, are unsuitable to participate in this trial.

[0034] 4.3 Criteria for Termination and Detachment 1. If the patient experiences an allergic reaction, serious adverse event, complication, or special physiological changes, they should not continue taking the medication; 2. The patient requests to withdraw or is lost to follow-up; 3. The study found that the drug had no practical value or that the clinical research protocol had major errors, making it difficult to evaluate the drug's efficacy.

[0035] 5. Probiotic composition intervention Sixty-eight patients were divided into a control group (n=27) receiving a probiotic composition containing only corn starch (350 mg / capsule); a treatment group (n=34) receiving a probiotic composition containing the probiotic composition of this invention (350 mg / capsule); three patients received Clostridium butyricum preparations; and four patients received Bacillus coagulans preparations. All four groups received one capsule twice a day for six months.

[0036] 6. Insomnia Severity Index Scale The Insomnia Severity Index (ISI) is a commonly used clinical tool for assessing insomnia. It is mainly used to quantify the severity of insomnia and its impact on daytime functioning. It is suitable for adults and has both clinical diagnostic reference and research evaluation value.

[0037] Table 1. Insomnia Severity Index (ISI) Scoring criteria: The total score is the sum of the scores for all 7 items, ranging from 0 to 28 points. Level 0: 0-7 points = insomnia with no clinical significance; Level 1: 8-14 points = mild insomnia; Level 2: 15-21 points = moderate insomnia; Level 3: 22-28 points = severe insomnia.

[0038] The Insomnia Severity Index (ISI) was used to rate 68 patients, and the resulting rating scale is shown in the table below: Table 2. Rating scores of 68 patients The following information was obtained by statistically analyzing the insomnia levels of the above 68 patients according to the scoring criteria: Table 3. Rating levels and percentages of 68 patients Based on the insomnia severity index scores of the patients mentioned above, and after understanding the patients' insomnia status, the patients were randomly divided into a probiotic composition control group, a probiotic composition treatment group, a Clostridium butyricum group, and a Bacillus coagulans group. Treatment was administered according to the prescribed drug intervention method. Follow-up visits were conducted and recorded at the first, third, and sixth months after administration. The results are shown in the table below: Table 4. Insomnia Severity Index (ISI) Scale for Probiotic Combinations Note: In the control group, 4 people dropped out during the third month of follow-up, and another 9 people dropped out during the sixth month of follow-up; Nine patients in the treatment group dropped out during the third month follow-up, and another eight dropped out during the sixth month follow-up.

[0039] The proportion of different rating levels in different groups was calculated based on the Insomnia Severity Index scale above, and the results are shown in the table below: Table 5. Statistical table of the proportion of different rating levels of probiotic compositions The experimental results are shown in Tables 4 and 5. Figure 1-6As shown, with the extension of follow-up time, the number of patients with grades 2 and 3 in the treatment group decreased. Before taking the probiotic composition, the proportion of grade 2 patients in the treatment group was relatively high, at 55.88%, meaning more than half of the patients had moderate insomnia; the proportion of patients with grade 1 (mild insomnia) was 29.41%; and the proportion of patients with grade 3 (severe insomnia) was 14.70%. With the extension of follow-up time, the proportion of patients with grade 0 continuously increased, while the proportion of patients with grades 2 and 3 decreased significantly, indicating a significant reduction in the severity of insomnia. This means that the probiotic composition of the present invention can effectively alleviate the severity of insomnia. The Insomnia Severity Index (ISI) suggests that the probiotic composition of the present invention can effectively reduce the impact of nighttime insomnia on daytime sleep while improving sleep duration and quality.

[0040] The ISI score table for three individuals who took Clostridium butyricum is shown in Table 4. Table 4 Clostridium butyricum ISI Scoring Table As shown in Table 4, the three patients had grade 2, grade 3, and grade 3 insomnia respectively at the first month follow-up. The follow-up results at the third and sixth months were consistent with those at the first month, indicating that Clostridium butyricum could not improve the patients' insomnia.

[0041] The ISI score table for 4 people who took Bacillus coagulans is shown in Table 5: Table 5. ISI Scoring Table for Bacillus coagulans As shown in Table 5, during the first month of follow-up, 2 people had grade 2 insomnia and 2 people had grade 3 insomnia. During the third month of follow-up, 3 people had grade 1 insomnia and 1 person had grade 2 insomnia. The results showed that Bacillus coagulans could improve insomnia. During the sixth month of follow-up, 3 people had recovered to grade 1 insomnia and 1 person's insomnia had been cured. That is, Bacillus coagulans can effectively reduce the severity of insomnia. In summary, the probiotic composition of this invention not only significantly improves insomnia symptoms but also synergistically improves sleep quality and related emotional states. Clostridium butyricum alone cannot improve sleep quality or sleep duration, and while Bacillus coagulans alone can improve sleep, the effect is not significant. The probiotic composition of this invention has clear technical effects and application value in the treatment and / or improvement of insomnia.

Claims

1. The use of a probiotic composition in the preparation of an agent to improve sleep disorders, characterized in that, The probiotic composition contains the following strains: Clostridium butyricum JSIM-MCB20040312 with accession number CGMCC No.1647 and Bacillus coagulans JSSW-LA-07 with accession number CGMCC No.2602.

2. The application as described in claim 1, characterized in that, The Clostridium butyricum and Bacillus coagulans were compounded at a live bacteria ratio of 1:5-30 CFU / g.

3. The application as described in claim 1, characterized in that, The concentrations of Clostridium butyricum and Bacillus coagulans were 1×10⁻⁶. 5 Up to 1×10 11 CFU / g.

4. The application as described in claim 1, characterized in that, The sleep disorders include insomnia and daytime functional impairment due to insufficient sleep.

5. The application as described in claim 1, characterized in that, Insomnia refers to a syndrome in which an individual experiences difficulty falling asleep, difficulty maintaining sleep, early awakening, or decreased sleep quality despite suitable sleep conditions and environment, and this condition persists and has a significant impact on daytime functioning.

6. The application as described in claim 1, characterized in that, The preparations include drugs, food, and food additives.

Images