Composition of probiotics and traditional chinese medicines for treating insomnia and use thereof

Abstract

A composition for treating insomnia, comprising a probiotic complex and traditional Chinese medicine components. The probiotic complex comprises Lactobacillus plantarum, Bifidobacterium lactis, Lactobacillus crispatus, Lactobacillus acidophilus, Lactobacillus rhamnosus, Bifidobacterium bifidum, Lactobacillus reuteri, Lactobacillus casei, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus fermentum, Lactobacillus gasseri, Lactococcus lactis, Bifidobacterium longum, Lactobacillus salivarius, Bifidobacterium breve, Streptococcus thermophilus, Lactobacillus paracasei and Lactobacillus helveticus. The traditional Chinese medicine components are semen ziziphi spinosae, bulbus lilii, poria, semen nelumbinis and mori fructus. The composition may further comprise prebiotics selected from L-arabinose, β-glucan, inulin, erythritol, lactitol, vitamin C and γ-aminobutyric acid. The composition can be used for preparing pharmaceutical or food formulations, such as solid beverages, chewable tablets or oral lozenges. Clinical observation shows that the composition has the effect of relieving insomnia.

Description

Probiotic and Traditional Chinese Medicine Combinations for Treating Insomnia and Their Applications Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a composition of probiotic complex and traditional Chinese medicine components and its use in treating or improving gastrointestinal inflammation, ulcers, Helicobacter pylori infection, constipation, and insomnia. Background Technology

[0002] Insomnia, commonly known as "sleeplessness" or "restless sleep," is a common health problem that affects the quality and quantity of sleep an individual. Insomnia can be classified into several types based on clinical manifestations and causes, such as difficulty falling asleep, difficulty maintaining sleep, early awakening, and mixed types. Its causes are complex and diverse, including psychological factors, physiological factors, environmental factors, and lifestyle habits. High-risk groups mainly include the elderly, women (especially menopausal women), those with a history or family history of mental illness, and people who work under long-term high pressure. Although insomnia itself does not directly cause death, long-term insomnia can significantly increase the risk of chronic diseases such as cardiovascular disease and diabetes, indirectly affecting quality of life and life expectancy.

[0003] Statistics show that there are over 300 million people suffering from insomnia in my country, and this number is increasing every year. Traditional treatments, such as taking Western medicine, are highly addictive and have many side effects. They can only work in terms of sedation and inducing sleep, but cannot be taken long-term. Other treatments, such as traditional Chinese medicine and physical therapy, have low efficacy and are time-consuming and laborious, causing a large number of people suffering from insomnia to suffer for a long time.

[0004] Probiotics play a comprehensive and significant role in human health, with clinical and practical applications in the immune system, digestive system, endocrine and metabolic system, nervous system regulation, and reproductive and urinary systems. People hope to treat insomnia by taking probiotics, but in conventional applications, problems such as poor bacterial activity or unclear function (no strain number), unscientific or singular strain combinations, low live bacteria count, and unscientific prebiotic formulations lead to insignificant efficacy of probiotic products, failing to address insomnia caused by various etiologies.

[0005] Current traditional Chinese medicine treatments mostly focus on local regulation, addressing symptoms such as liver fire disturbing the heart or stomach disharmony. However, the causes of insomnia are diverse and complex, and local regulation cannot fundamentally solve the problem. Furthermore, the human body has limitations in absorbing the large molecules of traditional Chinese medicine, making it difficult to maintain stable and noticeable effects.

[0006] Therefore, it is necessary to find a holistic solution by considering the various causes of insomnia and the need for a comprehensive approach. Summary of the Invention

[0007] In order to solve one of the above-mentioned technical problems in the prior art, the present invention provides a composition of probiotic complex and traditional Chinese medicine components and its use in improving insomnia.

[0008] In a first aspect, the present invention provides a composition comprising a probiotic complex and a traditional Chinese medicine component, wherein the probiotic complex comprises or is composed of the following strains: *Lactobacillus plantarum*, *Bifidobacterium lactis*, *Lactobacillus crispatus*, *Lactobacillus acidophilus*, *Lactobacillus rhamnosus*, *Bifidobacterium bifidum*, *Lactobacillus reuteri*, *Lactobacillus casei*, *Lactobacillus bulgaricus*, *Lactobacillus fermentum*, *Lactobacillus gersoni*, *Lactococcus lactis*, *Bifidobacterium longum*, and *Lactobacillus salivarius*. The herbal components include or are composed of the following herbs: jujube seed, lily bulb, poria cocos, lotus seed, and mulberry.

[0009] In some embodiments, the probiotic complex in the composition of the present invention comprises or is composed of the following bacterial species: *Lactobacillus plantarum*, *Bifidobacterium lactis*, *Lactobacillus crispatus*, *Lactobacillus acidophilus*, *Lactobacillus rhamnosus*, *Bifidobacterium bifidum*, *Lactobacillus reuteri*, *Lactobacillus casei*, *Lactobacillus bulgaricus*, *Lactobacillus fermentum*, *Lactobacillus gersoni*, *Lactococcus lactis*, *Bifidobacterium longum*, and *Lactobacillus salivarius*. *Lactobacillus salivarius*, *Bifidobacterium breve*, *Streptococcus thermophilus*, *Lactobacillus paracasei*, and *Lactobacillus helveticus*.

[0010] In some embodiments, the Lactobacillus plantarum is Lactobacillus plantarum LP9B, which was deposited on January 29, 2019, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 17243.

[0011] In some embodiments, the Bifidobacterium lactis is Bifidobacterium lactis Bb108, which was deposited on June 30, 2020, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 20165.

[0012] In some embodiments, the Lactobacillus crispatus is Lactobacillus crispatus LCP018, which was deposited on April 16, 2021, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 22190.

[0013] In some embodiments, the Lactobacillus acidophilus is Lactobacillus acidophilus LA5Y, which was deposited on January 29, 2019, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 17242.

[0014] In some embodiments, the Lactobacillus rhamnosus is Lactobacillus rhamnosus LR2Y, which was deposited on November 4, 2019, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 18800.

[0015] In some embodiments, the Bifidobacterium bifidum is Bifidobacterium bifidum CLME9, which was deposited on September 11, 2023, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 28421.

[0016] In some embodiments, the Lactobacillus reuteri is Lactobacillus reuteri B1r06, which was deposited on June 30, 2020, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 20164.

[0017] In some embodiments, the Lactobacillus casei is Lactobacillus casei BLc08, which was deposited on November 9, 2020, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 21128.

[0018] In some embodiments, the Lactobacillus bulgaricus is Lactobacillus bulgaricus, which was deposited on November 9, 2020, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 21127.

[0019] In some implementations, the Lactobacillus fermentum is Lactobacillus fermentum CCF33, which was deposited on June 7, 2022, at the China General Microbiological Culture Collection Center with accession number CGMCC No. 25015.

[0020] In some embodiments, the Lactobacillus gersoni is Lactobacillus gersoni LGD2, which was deposited on February 1, 2021, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 21785.

[0021] In some embodiments, the lactococcus lactis is Lactococcus lactis AQ001, which was deposited on June 7, 2022, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 25018.

[0022] In some embodiments, the Bifidobacterium longum is Bifidobacterium longum BL11, which was deposited on February 1, 2021, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 21783.

[0023] In some embodiments, the Lactobacillus salivarius is Lactobacillus salivarius LS18, which was deposited on April 16, 2021, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 22192.

[0024] In some embodiments, the Bifidobacterium breve is Bifidobacterium breve BB026, which was deposited on April 16, 2021, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 22189.

[0025] In some embodiments, the thermophilic streptococcus is Streptococcus thermophilus BStr06, which was deposited on November 9, 2020 at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 21129.

[0026] In some embodiments, the Lactobacillus paracasei is deposited on April 16, 2021, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 22191.

[0027] In some embodiments, the Lactobacillus helveticus is deposited on February 1, 2021, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 21782.

[0028] This invention utilizes the aforementioned 18 specific bacterial strains to form a probiotic complex, with a total live bacteria count exceeding 200 billion CFU / g. This complex effectively regulates the intestinal flora, treats and / or alleviates intestinal inflammation, ulcers, and constipation, inhibits and kills Helicobacter pylori, and improves sleep by regulating the axis relationship between intestinal and brain nerves. It also enhances the body's intestinal detoxification and metabolism capabilities and boosts immunity.

[0029] In some embodiments, the traditional Chinese medicine components in the composition of the present invention include or consist of the following traditional Chinese medicines: jujube seed, lily bulb, poria cocos, lotus seed, and mulberry.

[0030] In some embodiments, the composition of the present invention includes jujube seed.

[0031] Sour jujube seed has calming, soothing, heart-nourishing, and liver-tonifying effects. Its main components include sour jujube seed saponins, flavonoids, and trace elements such as zinc and calcium. It can regulate the central nervous system, exert a calming and sleep-promoting effect, and has a significant effect on improving insomnia caused by deficiency of heart and gallbladder qi.

[0032] In some embodiments, the composition of the present invention includes lily.

[0033] Lily has the effects of calming the mind and soothing the nerves, beautifying and moisturizing the skin, relieving dryness and cough. It can be used for symptoms such as mental confusion, insomnia and dreaminess, and depression. It can also treat insomnia caused by lung deficiency and heat.

[0034] In some embodiments, the composition of the present invention includes Poria cocos.

[0035] Poria cocos can promote diuresis, benefit the spleen and stomach, and calm the mind and nerves, thus having a remarkable effect on improving sleep quality. The polysaccharides, proteins, and trace elements contained in Poria cocos can soothe nerves, calm emotions, and make it easier for people to enter deep sleep, thereby improving sleep quality.

[0036] In some embodiments, the composition of the present invention includes lotus seeds.

[0037] Lotus seeds have the effects of strengthening the spleen and stopping diarrhea, tonifying the kidneys and consolidating essence, and nourishing the heart and calming the mind. Lotus seeds are rich in various nutrients, such as protein, vitamins, and minerals, which help nourish the body and balance endocrine function, thus achieving a calming and soothing effect. Some special components in lotus seeds, such as lotus seed alkaloids and lotus leaf alkaloids, have significant antioxidant and anti-inflammatory effects, helping to relieve brain fatigue, improve sleep quality, and have a good regulating effect on insomnia caused by excessive heart fire.

[0038] In some embodiments, the composition of the present invention includes mulberry.

[0039] Mulberries have the effects of nourishing yin and blood, promoting body fluid production, and moistening the intestines. They are primarily used to treat dizziness, vertigo, lower back pain, tinnitus, premature graying of hair, insomnia, excessive dreaming, thirst due to fluid depletion, diabetes, and constipation caused by liver and kidney deficiency and blood and essence deficiency. They are used to treat symptoms such as insomnia, dizziness, and vertigo. For insomnia sufferers, mulberries can help regulate the nervous system, relieve anxiety and irritability, thereby improving sleep quality and alleviating insomnia symptoms caused by qi deficiency.

[0040] The purpose of this invention is to provide a combination of probiotics and traditional Chinese medicine (TCM) that is both food and medicine, which can eliminate insomnia symptoms by treating and / or regulating intestinal diseases, constipation, and regulating brain nerves. The probiotic complex of this invention can promote the absorption of TCM, synergistically enhance the effects of TCM, and work in conjunction with TCM to adjust the body as a whole, achieving the goal of treating or improving insomnia holistically from multiple angles and addressing multiple causes.

[0041] In some embodiments, the compositions of the present invention further include prebiotics, said prebiotics including one or more of L-arabinose, β-glucan, inulin, erythritol, lactitol, vitamin C, and γ-aminobutyric acid.

[0042] In some embodiments, the composition of the present invention comprises L-arabinose.

[0043] L-arabinose is a prebiotic that promotes the growth of beneficial bacteria, including Bifidobacteria, and can help prevent constipation. L-arabinose selectively affects sucrase in the small intestine, thus inhibiting sucrose absorption. As a low-calorie sugar, L-arabinose can help prevent sleep disturbances caused by elevated blood sugar levels from sucrose intake.

[0044] In some embodiments, the compositions of the present invention comprise β-glucan.

[0045] β-glucan is an excellent nutrient source and effective growth factor for beneficial bacteria such as Bifidobacteria in the human gut, and can improve the digestive and absorptive functions of the human gut.

[0046] In some embodiments, the composition of the present invention comprises inulin.

[0047] Inulin can help control blood lipids, promote mineral absorption, prevent constipation, regulate gut microbiota, and improve gut health.

[0048] In some embodiments, the compositions of the present invention comprise erythritol.

[0049] Erythritol is a prebiotic that has a significant proliferative effect on Bifidobacteria in the gut. The short-chain fatty acids (acetic acid and lactic acid) produced by Bifidobacteria metabolism can effectively inhibit the growth of pathogens, thereby improving the body's immunity.

[0050] In some embodiments, the compositions of the present invention comprise lactitol.

[0051] Lactitol can promote intestinal peristalsis, accelerate the elimination of intestinal waste, prevent obesity, and promote the proliferation of probiotics.

[0052] In some embodiments, the composition of the present invention includes vitamin C.

[0053] Vitamin C has antioxidant properties, helps maintain immune function, and fights free radicals, which can improve immunity.

[0054] In some embodiments, the compositions of the present invention comprise γ-aminobutyric acid.

[0055] Gamma-aminobutyric acid (GABA) is an active amino acid that plays an important role in the energy metabolism of the human brain. It has a variety of physiological functions, including activating glucose metabolism in the brain, promoting acetylcholine synthesis, lowering blood ammonia, anticonvulsant effects, lowering blood pressure, improving brain function, stabilizing mental state, and promoting growth hormone secretion. GABA can also combat anxiety, depression, relieve stress, regulate mood, and promote the brain to fall asleep quickly and enter a deep sleep state as soon as possible.

[0056] In the composition of this invention, the synergistic effects of various probiotics and prebiotics are achieved to adjust the balance of intestinal flora, promote the transmission of information between the gut and brain nerves, and thus adjust human sleep. Probiotics regulate the brain by regulating hundreds of millions of neurons in the gut (vagus nerve, enteric nerve, spinal nerve), and establish an axial relationship between microorganisms, gut and brain, which can further improve insomnia caused by diseases or conditions such as gastrointestinal inflammation, ulcers, Helicobacter pylori infection, constipation, and heart-kidney disharmony.

[0057] In some embodiments, the composition of the present invention includes the probiotic complex, jujube seed, lily bulb, poria cocos, lotus seed, mulberry, L-arabinose, β-glucan, inulin, erythritol, lactitol, vitamin C, and γ-aminobutyric acid.

[0058] In some embodiments, the composition of the present invention comprises the following components in parts by weight: 2-6 parts probiotic complex, 2-5 parts jujube seed, 2-7 parts lily bulb, 2-6.5 parts poria cocos, 2-8 parts lotus seed, 2-8 parts mulberry, 2-7 parts L-arabinose, 1-4 parts β-glucan, 5-10 parts inulin, 6-10 parts erythritol, 6-10 parts lactitol, 0.5-3 parts vitamin C, and 2-4.5 parts γ-aminobutyric acid.

[0059] In some embodiments, the composition of the present invention comprises the following components in parts by weight: 3-5 parts probiotic complex, 3-5 parts jujube seed, 2-5 parts lily bulb, 3-5 parts Poria cocos, 2-5 parts lotus seed, 3-5 parts mulberry, 3.3-6.6 parts L-arabinose, 1-3 parts β-glucan, 6-8 parts inulin, 7-9 parts erythritol, 7-9 parts lactitol, 0.5-2 parts vitamin C, and 3-4.5 parts γ-aminobutyric acid.

[0060] In some embodiments, the composition of the present invention comprises the following components in parts by weight: 3 parts probiotic complex, 3 parts jujube seed, 3 parts lily bulb, 3 parts poria cocos, 2 parts lotus seed, 3 parts mulberry, 3.5 parts L-arabinose, 1.5 parts β-glucan, 7 parts inulin, 8 parts erythritol, 8 parts lactitol, 0.5 parts vitamin C, and 4.5 parts γ-aminobutyric acid.

[0061] In some embodiments, the viable counts of the strains in the probiotic complex of the present invention are as follows: *Lactobacillus plantarum* 850-950 billion CFU / g, *Bifidobacterium lactis* 50-150 billion CFU / g, *Lactobacillus curvatureii* 80-120 billion CFU / g, *Lactobacillus acidophilus* 100-180 billion CFU / g, *Lactobacillus rhamnosus* 50-150 billion CFU / g, *Bifidobacterium bifidum* 10-80 billion CFU / g, *Lactobacillus reuteri* 60-150 billion CFU / g, and *Lactobacillus casei* 60-150 billion CFU / g. Lactobacillus bulgaricus 10-60 billion CFU / g, Lactobacillus fermentum 60-100 billion CFU / g, Lactobacillus gasseri 10-60 billion CFU / g, Lactococcus lactis 10-60 billion CFU / g, Bifidobacterium longum 20-80 billion CFU / g, Lactobacillus salivarius 100-150 billion CFU / g, Bifidobacterium breve 10-80 billion CFU / g, Streptococcus thermophilus 50-100 billion CFU / g, Lactobacillus paracasei 120-160 billion CFU / g, Lactobacillus helveticus 50-100 billion CFU / g.

[0062] In some embodiments, the viable counts of the strains in the probiotic complex of the present invention are as follows: *Lactobacillus plantarum* 90-93 billion CFU / g, *Bifidobacterium lactis* 10-13 billion CFU / g, *Lactobacillus curvatureii* 8-11 billion CFU / g, *Lactobacillus acidophilus* 13-16 billion CFU / g, *Lactobacillus rhamnosus* 10-13 billion CFU / g, *Bifidobacterium bifidum* 2-5 billion CFU / g, *Lactobacillus reuteri* 10-13 billion CFU / g, and *Lactobacillus casei* 10-13 billion CFU / g. U / g, Lactobacillus bulgaricus 10-40 billion CFU / g, Lactobacillus fermentum 60-90 billion CFU / g, Lactobacillus gasseri 10-40 billion CFU / g, Lactococcus lactis 10-40 billion CFU / g, Bifidobacterium longum 30-60 billion CFU / g, Lactobacillus salivarius 100-130 billion CFU / g, Bifidobacterium breve 30-60 billion CFU / g, Streptococcus thermophilus 50-80 billion CFU / g, Lactobacillus paracasei 120-150 billion CFU / g, Lactobacillus helveticus 50-80 billion CFU / g.

[0063] In some embodiments, the viable counts of the strains in the probiotic complex of the present invention are as follows: *Lactobacillus plantarum* 90 billion CFU / g, *Bifidobacterium lactis* 10 billion CFU / g, *Lactobacillus curvatureii* 8 billion CFU / g, *Lactobacillus acidophilus* 13 billion CFU / g, *Lactobacillus rhamnosus* 10 billion CFU / g, *Bifidobacterium bifidum* 2 billion CFU / g, *Lactobacillus reuteri* 10 billion CFU / g, *Lactobacillus casei* 10 billion CFU / g, *Lactobacillus bulgaricus* 1 billion CFU / g, *Lactobacillus fermentum* 6 billion CFU / g, *Lactobacillus gasseri* 1 billion CFU / g, *Lactococcus lactis* 1 billion CFU / g, *Bifidobacterium longum* 3 billion CFU / g, *Lactobacillus salivarius* 10 billion CFU / g, *Bifidobacterium breve* 3 billion CFU / g, *Streptococcus thermophilus* 5 billion CFU / g, *Lactobacillus paracasei* 12 billion CFU / g, and *Lactobacillus helveticus* 5 billion CFU / g.

[0064] In some embodiments, the total number of live bacteria in the probiotic complex of the present invention is above 200 billion CFU / g, above 210 billion CFU / g, or above 220 billion CFU / g.

[0065] In a second aspect, the present invention provides a formulation made from the composition of the first aspect of the present invention.

[0066] The formulation of the present invention can be prepared by the following method: according to the composition of the first aspect, a probiotic complex is taken in parts by weight and prepared into probiotic powder by encapsulation technology; each traditional Chinese medicine component is taken in parts by weight and a water extract of traditional Chinese medicine is obtained by water extraction and prepared into powder; each prebiotic is taken in parts by weight and prepared into powder; the probiotic powder, the water extract powder of traditional Chinese medicine, and the prebiotic powder are dried, the dried powders of each component are mixed, and the formulation is prepared into solid beverage, chewable tablet or oral lozenge according to conventional formulation methods.

[0067] The embedding technique can be implemented using conventional techniques in this field.

[0068] In some embodiments, the encapsulation can be performed as follows: a protective agent is evenly sprayed onto the bacterial sludge in an emulsification tank, stirred, and then encapsulated. The protective agent can be, for example, lactic acid bacteria extracellular polysaccharides extracted from high-extracellular polysaccharide-producing strains such as *Bifidobacterium longum*, *Bifidobacterium lactis*, *Streptococcus thermophilus*, and *Lactobacillus casei*.

[0069] The water extract of the traditional Chinese medicine can be prepared using conventional techniques in the field or obtained commercially.

[0070] In some embodiments, the aqueous extract of the traditional Chinese medicine can be prepared as follows: Take jujube seed, lily bulb, poria cocos, lotus seed, and mulberry by weight, pulverize them, soak them in 6-8 times their volume of water for 30-60 minutes, and extract them 2-3 times in an extraction tank. The first extraction is carried out by boiling for 60-80 minutes, and the second extraction is carried out by boiling in 4-5 times their volume of water for 30-50 minutes. Combine the two filtrates, concentrate them to one times the volume of the medicinal materials, sieve, and dry them into a fine powder to obtain the final product.

[0071] In some embodiments, the aqueous extract of traditional Chinese medicine can be obtained by mixing commercially available powders of aqueous extracts of various traditional Chinese medicine components.

[0072] The formulation of this invention can be taken by dissolving in warm water or by direct oral administration.

[0073] Thirdly, the present invention also provides the use of the compositions or preparations described herein in the preparation of medicaments and / or foods for treating or improving gastrointestinal inflammation, ulcers, Helicobacter pylori infection, constipation, and insomnia.

[0074] In some embodiments, the composition or formulation of the present invention is used in the preparation of a medicament for treating gastrointestinal inflammation, ulcers, Helicobacter pylori infection, constipation, and insomnia.

[0075] In some embodiments, the compositions or preparations of the present invention are used in the preparation of foods for improving gastrointestinal inflammation, ulcers, Helicobacter pylori infection, constipation, and insomnia.

[0076] In some embodiments, the composition or formulation of the present invention is used in the preparation of a medicament for treating insomnia.

[0077] In some embodiments, the composition or formulation described herein is used in the preparation of a food for improving insomnia.

[0078] In some implementations, the insomnia includes insomnia caused by gastrointestinal inflammation, ulcers, Helicobacter pylori infection, constipation, heart-kidney disharmony, etc.

[0079] The composition of the present invention includes a probiotic complex and traditional Chinese medicine components that are both food and medicine, as well as prebiotics that promote the growth of probiotics and serve as their nutrient source. The probiotic complex contains more than 200 billion CFU / g of active probiotics. By combining probiotics with traditional Chinese medicine that is both food and medicine, and supplementing them with appropriate prebiotics, it is possible to treat or improve insomnia caused by spleen and stomach disharmony, liver qi stagnation, heart fire excess, yin deficiency and fire excess, heart and kidney disharmony, etc., which are caused by emotions, stress, diseases, etc.

[0080] The composition of the present invention, while the various components work synergistically to treat or improve insomnia, also promotes the absorption of various medicinal and edible herbs and other ingredients, maximizing their effects. At the same time, the various components can play an overall synergistic role, achieving the goal of treating or improving insomnia from multiple angles and multiple causes. Detailed Implementation

[0081] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. The specific embodiments described herein are for illustrative purposes only and are not intended to limit the invention in any way. Furthermore, descriptions of well-known structures and techniques are omitted in the following description to avoid unnecessarily obscuring the concepts of this disclosure. Such structures and techniques have also been described in many publications.

[0082] definition

[0083] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly used in the field to which this invention pertains. For the purposes of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural forms, and vice versa.

[0084] Unless otherwise specified, all references to quantity in this document include their plural forms.

[0085] As used herein, the term "about" indicates a range of ±20% of the following value. In some embodiments, the term "about" indicates a range of ±10% of the following value. In some embodiments, the term "about" indicates a range of ±5% of the following value.

[0086] The numerical ranges mentioned in this article include the extreme values ​​and any values ​​in between. For example, the weight range is 5-10 parts, including 5, 6, 7, 8, 9, 10 parts and any values ​​in between; the viable count of the strain is 85-95 billion CFU / g, including 85 billion CFU / g, 86 billion CFU / g, 87 billion CFU / g, 88 billion CFU / g, 89 billion CFU / g, 90 billion CFU / g, 91 billion CFU / g, 92 billion CFU / g, 93 billion CFU / g, 94 billion CFU / g, 95 billion CFU / g and any values ​​in between; and so on.

[0087] The following embodiments are provided to aid in understanding the present invention. However, it should be understood that these embodiments are for illustrative purposes only and do not constitute any limitation. The actual scope of protection of the present invention is set forth in the claims. It should be understood that any modifications and changes can be made without departing from the spirit of the present invention.

[0088] Unless otherwise specified, the Chinese herbal medicines and prebiotics mentioned in the following examples are also commercially available.

[0089] Example 1.

[0090] This embodiment provides a probiotic composition comprising the following components in parts by weight:

[0091] Three portions of probiotic complex (Lactobacillus plantarum 90 billion CFU / g, Bifidobacterium lactis 10 billion CFU / g, Lactobacillus curvatureii 8 billion CFU / g, Lactobacillus acidophilus 13 billion CFU / g, Lactobacillus rhamnosus 10 billion CFU / g, Bifidobacterium bifidum 2 billion CFU / g, Lactobacillus reuteri 10 billion CFU / g, Lactobacillus casei 10 billion CFU / g, Lactobacillus bulgaricus 1 billion CFU / g). Lactobacillus fermentum 6 billion CFU / g, Lactobacillus gasseri 1 billion CFU / g, Lactococcus lactis 1 billion CFU / g, Bifidobacterium longum 3 billion CFU / g, Lactobacillus salivarius 10 billion CFU / g, Bifidobacterium breve 3 billion CFU / g, Streptococcus thermophilus 5 billion CFU / g, Lactobacillus paracasei 12 billion CFU / g, Lactobacillus helveticus 5 billion CFU / g; total effective viable count 200 billion CFU / g;

[0092] 3 parts jujube seed, 3 parts lily bulb, 3 parts poria cocos, 2 parts lotus seed, 3 parts mulberry, 3.5 parts L-arabinose, 1.5 parts β-glucan, 7 parts inulin, 8 parts erythritol, 8 parts lactitol, 0.5 parts vitamin C, and 4.5 parts γ-aminobutyric acid.

[0093] The above 18 kinds of bacterial powders were processed using an encapsulation technique: the extracellular polysaccharides of lactic acid bacteria were evenly sprayed onto the bacterial mud in an emulsification tank, stirred, encapsulated and protected, and dried to produce probiotic powder.

[0094] The above five Chinese herbal medicine components were processed by water decoction extraction: Ziziphus jujuba seed, lily bulb, Poria cocos, lotus seed, and mulberry were weighed according to their weight proportions, soaked in 6 times their volume of water for 40 minutes, and then extracted twice in an extraction tank. The first extraction was carried out by boiling for 70 minutes, and the second extraction was carried out by boiling in 5 times their volume of water for 40 minutes. The two filtrates were combined, concentrated to 1 times the volume of the medicinal materials, filtered, and dried into a fine powder to obtain the final product.

[0095] The probiotic powder, the water extract powder of traditional Chinese medicine, and the prebiotic component powder are mixed, dried, and made into a solid beverage preparation, 5g / dose.

[0096] Example 2.

[0097] This embodiment provides a probiotic composition comprising the following components in parts by weight:

[0098] Four portions of probiotic complex (Lactobacillus plantarum 91 billion CFU / g, Bifidobacterium lactis 11 billion CFU / g, Lactobacillus curvatureii 10 billion CFU / g, Lactobacillus acidophilus 10 billion CFU / g, Lactobacillus rhamnosus 11 billion CFU / g, Bifidobacterium bifidum 3 billion CFU / g, Lactobacillus reuteri 11 billion CFU / g, Lactobacillus casei 8 billion CFU / g, Lactobacillus bulgaricus 2 billion CFU / g) Lactobacillus fermentum 6 billion CFU / g, Lactobacillus gasseri 1 billion CFU / g, Lactococcus lactis 1 billion CFU / g, Bifidobacterium longum 2 billion CFU / g, Lactobacillus salivarius 10 billion CFU / g, Bifidobacterium breve 1 billion CFU / g, Streptococcus thermophilus 5 billion CFU / g, Lactobacillus paracasei 12 billion CFU / g, Lactobacillus helveticus 5 billion CFU / g; total effective viable count 200 billion CFU / g;

[0099] 3 parts jujube seed, 5 parts lily bulb, 4 parts poria cocos, 4 parts lotus seed, 5 parts mulberry, 4 parts L-arabinose, 2 parts β-glucan, 6 parts inulin, 7 parts erythritol, 9 parts lactitol, 1 part vitamin C, and 4 parts γ-aminobutyric acid.

[0100] Using the same encapsulation and water extraction methods as in Example 1, the obtained component powders were mixed, dried, and prepared into a solid beverage formulation, 5g / dose.

[0101] Example 3.

[0102] This embodiment provides a probiotic composition comprising the following components in parts by weight:

[0103] Three portions of probiotic complex (Lactobacillus plantarum 93 billion CFU / g, Bifidobacterium lactis 10 billion CFU / g, Lactobacillus curvatureii 8 billion CFU / g, Lactobacillus acidophilus 12 billion CFU / g, Lactobacillus rhamnosus 10 billion CFU / g, Bifidobacterium bifidum 3 billion CFU / g, Lactobacillus reuteri 10 billion CFU / g, Lactobacillus casei 10 billion CFU / g, Lactobacillus bulgaricus 2 billion CFU / g). Lactobacillus fermentum 6 billion CFU / g, Lactobacillus gasseri 1 billion CFU / g, Lactococcus lactis 1 billion CFU / g, Bifidobacterium longum 6 billion CFU / g, Lactobacillus salivarius 10 billion CFU / g, Bifidobacterium breve 6 billion CFU / g, Streptococcus thermophilus 5 billion CFU / g, Lactobacillus paracasei 11 billion CFU / g, Lactobacillus helveticus 6 billion CFU / g; total effective viable count 210 billion CFU / g;

[0104] 4 parts jujube seed, 3 parts lily bulb, 5 parts poria cocos, 2 parts lotus seed, 4 parts mulberry, 6.6 parts L-arabinose, 2 parts β-glucan, 6 parts inulin, 9 parts erythritol, 8 parts lactitol, 0.5 parts vitamin C, and 3 parts γ-aminobutyric acid.

[0105] Using the same encapsulation and water extraction methods as in Example 1, the obtained component powders were mixed, dried, and made into chewable tablets, 5g / dose.

[0106] Example 4.

[0107] This embodiment provides a probiotic composition comprising the following components in parts by weight:

[0108] Probiotic complex (5 portions): Lactobacillus plantarum 85 billion CFU / g, Bifidobacterium lactis 6 billion CFU / g, Lactobacillus cremastrae 8 billion CFU / g, Lactobacillus acidophilus 10 billion CFU / g, Lactobacillus rhamnosus 6 billion CFU / g, Bifidobacterium bifidum 2 billion CFU / g, Lactobacillus reuteri 6 billion CFU / g, Lactobacillus casei 6 billion CFU / g, Lactobacillus bulgaricus 2 billion CFU / g, Lactobacillus fermentum The total number of viable bacteria was 10 billion CFU / g, including Lactobacillus gasseri 6 billion CFU / g, Lactococcus lactis 6 billion CFU / g, Bifidobacterium longum 8 billion CFU / g, Lactobacillus salivarius 15 billion CFU / g, Bifidobacterium breve 8 billion CFU / g, Streptococcus thermophilus 10 billion CFU / g, Lactobacillus paracasei 16 billion CFU / g, and Lactobacillus helveticus 10 billion CFU / g; the total number of viable bacteria was 220 billion CFU / g.

[0109] 3 parts jujube seed, 2 parts lily bulb, 3 parts poria cocos, 3 parts lotus seed, 3 parts mulberry, 5 parts L-arabinose, 2 parts β-glucan, 8 parts inulin, 7 parts erythritol, 8 parts lactitol, 0.5 parts vitamin C, and 3 parts γ-aminobutyric acid.

[0110] Using the same encapsulation and water extraction methods as in Example 1, the obtained component powders were mixed, dried, and made into oral lozenges, 5g / dose.

[0111] Comparative Example 1.

[0112] This comparative example provides a probiotic composition comprising the following components in parts by weight:

[0113] Lactobacillus plantarum 40 billion CFU / g, Bifidobacterium lactis 5 billion CFU / g, Lactobacillus curvatureii 8 billion CFU / g, Lactobacillus acidophilus 5 billion CFU / g, Lactobacillus rhamnosus 5 billion CFU / g, Bifidobacterium bifidum 1 billion CFU / g, Lactobacillus reuteri 6 billion CFU / g, Lactobacillus casei 6 billion CFU / g, Lactobacillus fermentum 6 billion CFU / g, Lactobacillus gasseri 1 billion CFU / g, Lactococcus lactis 1 billion CFU / g, Bifidobacterium longum 2 billion CFU / g, Lactobacillus salivarius 6 billion CFU / g, Bifidobacterium breve 3 billion CFU / g, Streptococcus thermophilus 5 billion CFU / g; Total effective viable count 100 billion CFU / g;

[0114] 10 parts of jujube seed, 3 parts of lily bulb, 3 parts of poria cocos, 2 parts of lotus seed, 3 parts of mulberry, 2 parts of longan pulp, 2 parts of L-arabinose, 2 parts of galactooligosaccharide, 7 parts of inulin, 8 parts of erythritol, and 6 parts of γ-aminobutyric acid.

[0115] Using the same encapsulation and water extraction methods as in Example 1, the obtained component powders were mixed, dried, and prepared into a solid beverage formulation, 5g / dose.

[0116] Comparative Example 2.

[0117] This comparative example provides a probiotic composition comprising the following components in parts by weight:

[0118] Lactobacillus plantarum 42 billion CFU / g, Bifidobacterium lactis 6 billion CFU / g, Lactobacillus curvatureii 9 billion CFU / g, Lactobacillus acidophilus 8 billion CFU / g, Lactobacillus rhamnosus 8 billion CFU / g, Bifidobacterium bifidum 2 billion CFU / g, Lactobacillus reuteri 10 billion CFU / g, Lactobacillus casei 10 billion CFU / g, Lactobacillus bulgaricus 2 billion CFU / g, Lactobacillus fermentum 8 billion CFU / g, Lactobacillus gasseri 2 billion CFU / g, Lactococcus lactis 2 billion CFU / g, Bifidobacterium longum 3 billion CFU / g, Lactobacillus salivarius 10 billion CFU / g, Bifidobacterium breve 3 billion CFU / g, Streptococcus thermophilus 5 billion CFU / g; Total effective viable count 130 billion CFU / g;

[0119] 7 parts of jujube seed, 3 parts of lily bulb, 3 parts of poria cocos, 2 parts of lotus seed, 3 parts of mulberry, 3 parts of L-arabinose, 3 parts of galactooligosaccharide, 3 parts of monk fruit glycoside, 6 parts of inulin, and 5 parts of γ-aminobutyric acid.

[0120] Using the same encapsulation and water extraction methods as in Example 1, the obtained component powders were mixed, dried, and prepared into a solid beverage formulation, 5g / dose.

[0121] Comparative Example 3.

[0122] This comparative example provides a probiotic composition comprising the following components in parts by weight:

[0123] Lactobacillus plantarum 52 billion CFU / g, Bifidobacterium lactis 10 billion CFU / g, Lactobacillus curvatureii 8 billion CFU / g, Lactobacillus acidophilus 13 billion CFU / g, Lactobacillus rhamnosus 10 billion CFU / g, Bifidobacterium bifidum 2 billion CFU / g, Lactobacillus reuteri 11 billion CFU / g, Lactobacillus casei 11 billion CFU / g, Lactobacillus bulgaricus 2 billion CFU / g, Lactobacillus fermentum 6 billion CFU / g, Lactobacillus gasseri 1 billion CFU / g, Lactococcus lactis 1 billion CFU / g, Bifidobacterium longum 3 billion CFU / g, Lactobacillus salivarius 12 billion CFU / g, Bifidobacterium breve 3 billion CFU / g, Streptococcus thermophilus 5 billion CFU / g; Total effective viable count 150 billion CFU / g;

[0124] 6 parts of jujube seed, 3 parts of lily bulb, 3 parts of poria cocos, 2 parts of lotus seed, 3 parts of mulberry, 3 parts of L-arabinose, 2 parts of galactooligosaccharide, 8 parts of erythritol, 7 parts of inulin, 7 parts of lactitol, and 3 parts of γ-aminobutyric acid.

[0125] Using the same encapsulation and water extraction methods as in Example 1, the obtained component powders were mixed, dried, and prepared into a solid beverage formulation, 5g / dose.

[0126] Comparative Example 4.

[0127] Cinnabar Tranquilizing Pills (Jilin Xinhui Pharmaceutical Co., Ltd.), 9g / pill.

[0128] Experimental Example 1. Isolation and Screening of Strains

[0129] 1. Separation and screening methods

[0130] Samples: Yogurt, kimchi, human intestinal isolate, human saliva.

[0131] Method: Weigh an appropriate amount of sample into sterile physiological saline (with an appropriate amount of glass beads added) as the stock solution, shake for 10 min to mix thoroughly, and perform 10... -2 ~10 -7 Serial dilutions were performed. The cultures were plated onto Lactobacillus MRS medium and Bifidobacterium BL selective medium, with two replicates for each dilution. The cultures were incubated anaerobicly at 37°C for 48 hours, and colony morphology was observed, numbered, and recorded. Gram-positive, catalase-negative non-spore-forming bacilli were cryopreserved using catalase, gas production, and capsular spore tests. Simultaneously, they were deposited at the China General Microbiological Culture Collection Center.

[0132] 2. Microbiological characteristics

[0133] Lactobacillus plantarum (CGMCC No. 17243):

[0134] On MRS plates, the bacteria are white, round, smooth, moist, with regular edges, and raised, ranging from 0.5 mm to 2 mm in diameter. They are Gram-positive, catalase-negative, and appear as short rods under a microscope.

[0135] Bifidobacterium lactis (CGMCC No. 20165):

[0136] Anaerobic culture on bifidobacteria medium produces white, round, moist, opaque colonies with neat edges. They are Gram-positive and catalase-negative. Under a microscope, the bacteria appear as rod-shaped cells with swollen ends.

[0137] Lactobacillus curvatureii (CGMCC No. 22190):

[0138] The colonies are white, with irregular edges, flaky, and convex in the center. They can utilize mannose, galactose, glucose, maltose, melibiose, raffinose, ribose, sucrose, trehalose, and fructose. They are Gram-positive and appear as slender rods, slightly bent into chains, under a microscope.

[0139] Lactobacillus acidophilus (CGMCC No. 17242):

[0140] Small, net-like, raised, rough-surfaced colonies with curled edges form on the agar plate. Gram-positive, appearing as bacilli under a microscope.

[0141] Lactobacillus rhamnosus (CGMCC No. 18800):

[0142] On agar plates, they appear as white, round, smooth, moist colonies with neat, raised edges. They cannot utilize lactose but can metabolize monosaccharides; they are Gram-positive and appear as bacilli under a microscope.

[0143] Bifidobacterium bifidum (CGMCC No. 28421):

[0144] Gram-positive, rod-shaped cells, arranged singly, in pairs, in V-shapes, or in clusters. Staining is usually irregular. Non-motile, does not produce spores, acid-fast stain negative; fermentation products are mainly acetic acid and lactic acid, no CO2 produced. Does not produce butyric acid or propionic acid. Catalase-negative, strictly anaerobic, ferments glucose to produce lactic acid.

[0145] Lactobacillus reuteri (CGMCC NO.20164):

[0146] Colonies grown on MRS plates are all round, milky white, and smooth; Gram-positive bacteria, when observed under a microscope, appear as slightly curved rods.

[0147] Lactobacillus casei (CGMCC No. 21128):

[0148] It forms round, moist colonies with a diameter of 0.5-2 mm, white (sometimes slightly yellow), Gram-positive, non-spore-forming, non-motile, and varying in length, often in chains.

[0149] Lactobacillus bulgaricus (CGMCC No. 21127):

[0150] Gram-positive, thick and long rod-shaped, slightly curved in chains, non-flagellated, non-spore-forming, colonies grayish-white with irregular edges, viscous, and a raised center; chemoheterotrophic, facultative anaerobic, does not liquefy gelatin, and can utilize fructose, glucose, and lactose. Catalase-negative, oxidase-negative, acid-tolerant, thermophilic, with a growth temperature of 15-40℃.

[0151] Lactobacillus fermentum (CGMCC No. 25015):

[0152] Colonies are round with neat edges and a milky-white color. Cells are rod-shaped. They do not form spores and are Gram-positive. They are non-motile and lack a capsule. They are facultative anaerobic. The optimum temperature is 36℃.

[0153] Lactobacillus gasseri (CGMCC No. 21785):

[0154] The colony morphology of the strain on MRS solid medium is round, with a diameter of 0.5-0.75 mm and a rough surface. Under a microscope, it appears as a Gram-positive short rod-shaped bacterium.

[0155] Lactococcus lactis (CGMCC No. 25018):

[0156] The cells are spherical or oval, Gram-positive, facultatively anaerobic, do not produce capsules or spores, have complex nutritional requirements, and the optimal growth temperature is 30℃.

[0157] Bifidobacterium longum (CGMCC No. 21783):

[0158] Gram-positive, tadpole-shaped with one end enlarged, appearing in clusters, short cells, usually rod-shaped, without flagella, without spores, colonies are white, shiny, with neat, round edges, and a raised center, chemoheterotrophic, anaerobic.

[0159] Lactobacillus salivarius (CGMCC No. 22192):

[0160] Gram-positive, short rods, appearing in clusters, non-flagellated, non-spore-forming, colonies round, milky white, raised, with neat edges, chemoheterotrophic, facultative anaerobic, and does not liquefy gelatin.

[0161] Bifidobacterium breve (CGMCC No. 22189):

[0162] The cells are short, usually rod-shaped, Gram-positive, anaerobic, produce acid from glucose, are catalase-negative, and produce α-galactosidase.

[0163] Streptococcus thermophilus (CGMCC No. 21129):

[0164] Colonies are small, round, raised, and shiny; cells are spherical or oval, occurring in pairs or chains. Gram-positive, catalase-negative. Non-spore-forming, non-capsulated, and non-motile.

[0165] Lactobacillus paracasei (CGMCC No. 22191):

[0166] Gram-positive, catalase-negative. It grows readily on simple nutrient media, and its colonies may be smooth (S), low-convex, moist, glossy, entire, gray, or rough (R).

[0167] Lactobacillus helveticus (CGMCC No. 21782):

[0168] Gram-positive bacteria are probiotics. They are rod-shaped, non-flagellated, non-spore-forming, with round, milky-white colonies and regular edges. They are chemoheterotrophic and facultative anaerobic.

[0169] 3. Proliferation characteristics

[0170] Lactobacillus plantarum (CGMCC No. 17243):

[0171] It is an aerobic or facultative anaerobic, or facultative heterotrophic fermenting lactic acid bacteria, with an optimal growth temperature of 30-37℃ and an optimal pH of around 6.5. The optimal nitrogen source combination is 10 g / L yeast extract, 10 g / L peptone, and 2 g / L triammonium citrate.

[0172] Bifidobacterium lactis (CGMCC No. 20165):

[0173] Anaerobic bacteria have an optimal growth temperature of 37-41℃; an optimal fermentation temperature of 35-40℃; a minimum growth temperature of 25-28℃; a maximum growth temperature of 43-45℃; and an initial growth pH of 6.7-7.0. They do not grow in environments with a pH below 4.5-5.0 or a pH above 8.0-8.5.

[0174] Lactobacillus curvatureii (CGMCC No. 22190):

[0175] It is a facultative anaerobe, produces acid, and prefers warm temperatures, with a growth temperature of 15-40℃ and an optimal growth temperature of 37℃. It tolerates pH 6.5 well and exhibits strong tolerance to acids and bile salts, even growing slowly in an acidic environment with a pH of 3.5. It has complex nutritional requirements and can degrade various carbohydrates, producing L- and D-lactic acid isomers.

[0176] Lactobacillus acidophilus (CGMCC No. 17242):

[0177] It is anaerobic or facultative anaerobic. It is highly acid-tolerant and grows well in slightly acidic environments, with an optimal growth pH of around 5.5-6.0. It is relatively sensitive to temperature, failing to grow below 20℃ and exhibiting poor heat tolerance, generally growing best at around 37℃. It can undergo homofermentation using glucose, fructose, lactose, and sucrose.

[0178] Lactobacillus rhamnosus (CGMCC No. 18800):

[0179] It is a facultative anaerobic bacterium that can grow in a relatively wide pH range of 4.5-5.5, with an optimal pH of 5.5-6.2 and an optimal growth temperature of around 37℃. The fermentation temperature range is 30℃-40℃, with an optimal fermentation temperature of around 35℃.

[0180] Bifidobacterium bifidum (CGMCC No. 28421):

[0181] An obligate anaerobic bacterium, with an optimal growth temperature of 37-38℃ and an optimal pH of 6.0-7.0. It requires carbon dioxide for growth. It has strict nutritional requirements, needing to ferment sugars, casein hydrolysate, various amino acids, and vitamins. It can ferment glucose, fructose, lactose, and galactose.

[0182] Lactobacillus reuteri (CGMCC NO.20164):

[0183] The optimal growth temperature is 36-40℃, the optimal fermentation temperature is 35-40℃, the minimum growth temperature is 25-28℃, the maximum growth temperature is 43-45℃, and the initial growth pH is 6.7-7.0. It generally does not grow in environments with a pH below 4.5-5.0 or a pH above 8.0-8.5.

[0184] Lactobacillus casei (CGMCC No. 21128):

[0185] Microaerophilic bacteria have low oxygen requirements and can survive and reproduce in relatively low-oxygen environments. The suitable growth temperature is 30-37℃, with the optimum temperature being 37℃. They can grow in environments with a pH value of 4.5-9.0, with the optimum pH range being 6.2-6.6. They have good acid resistance and bile resistance.

[0186] Lactobacillus bulgaricus (CGMCC No. 21127):

[0187] Microaerophilic or anaerobic bacteria, *Lactobacillus bulgaricus* grows best in anaerobic environments. The optimal growth temperature is around 40℃; it cannot grow above 50℃ or below 20℃. *Lactobacillus bulgaricus* can grow and reproduce well within a temperature range of 37℃-45℃, but it has poor heat tolerance. It thrives in acidic environments, with an optimal pH of 4.6-5.4. Within this pH range, the bacteria maintain optimal physiological activity.

[0188] Lactobacillus fermentum (CGMCC No. 25015):

[0189] Facultative anaerobic bacteria have a wide suitable temperature range for growth, generally between 25℃ and 40℃. Specifically, *Lactobacillus fermentum* grows best in the temperature range of 30℃ to 37℃, with a suitable pH range of 5.0 to 6.5. Within this pH range, the growth and reproduction of *Lactobacillus fermentum* are most active.

[0190] Lactobacillus gasseri (CGMCC No. 21785):

[0191] This facultative heterogeneous acidophilic strain can grow in both aerobic and anaerobic environments. The optimal growth pH is 5-7, the optimal growth temperature is 30-37℃, and the optimal inoculum size is 0.5%-2% (v / v). It can grow in 0.1% bile salts and in 5% NaCl.

[0192] Lactococcus lactis (CGMCC No. 25018):

[0193] Facultative anaerobic, with an optimal growth temperature of 30℃, it can survive in the range of 20-45℃, but the specific temperature range may vary depending on the strain. The optimal growth pH is 5.5-6.0, and it has strong acid resistance, enabling it to reproduce in acidic environments where other lactic acid bacteria cannot grow.

[0194] Bifidobacterium longum (CGMCC No. 21783):

[0195] Obligate anaerobic bacteria. MRS cultures show good growth under concentrated anaerobic conditions, with an optimal growth temperature of 37-38℃ and an optimal pH of 6.0-7.0. They can ferment glucose, fructose, sucrose, etc., to produce lactic acid and acetic acid. Some strains exhibit weaker fermentation performance in milk but stronger ability to hydrolyze casein.

[0196] Lactobacillus salivarius (CGMCC No. 22192):

[0197] This is a facultative heteroacidic strain that thrives in facultative or absolute anaerobic conditions. The optimal growth temperature is 30-43℃, and the optimal pH is 5.0-5.5 or lower. The most suitable growth temperature is around 37℃. It grows better in anaerobic environments, produces lactic acid, and is acid-resistant, tolerating pH up to 2.5 and bile salts up to 0.4% for 4 hours.

[0198] Bifidobacterium breve (CGMCC No. 22189):

[0199] For obligate anaerobes, the key to cultivation and viable cell counting is providing an anaerobic environment with low redox potential. The optimal growth temperature is 37℃-41℃, the minimum is 25℃-28℃, and the maximum is 43℃-45℃. The initial optimal pH is 6.5-7.0; growth does not occur at pH 4.5-5.0 or pH 8.0-8.5.

[0200] Streptococcus thermophilus (CGMCC No. 21129):

[0201] Facultative anaerobic bacteria have an optimal growth temperature of around 40°C, but can also grow at 45°C under laboratory conditions. Furthermore, studies have indicated that *Streptococcus thermophilus* can also grow at 37°C. Growth is optimal in a culture medium with a pH of 7.0. They can utilize sugars such as galactose, glucose, fructose, lactose, and sucrose for growth.

[0202] Lactobacillus paracasei (CGMCC No. 22191):

[0203] It is a facultative anaerobe, with a growth temperature range of 15-45℃, an optimum temperature of 37℃, and is acid-resistant, tolerating pH up to 2.5, with an optimum pH of 5.3. It primarily produces L-lactic acid during glucose fermentation.

[0204] Lactobacillus helveticus (CGMCC No. 21782):

[0205] These are aerobic or microaerophilic bacteria, with a suitable temperature range of 30-45℃ and an optimum temperature of 37℃. They can utilize sugars such as glucose, fructose, maltose, and lactose in a 1:1 ratio as carbon sources, and produce DL-lactic acid through homofermentation. They cannot utilize disaccharides such as sucrose and rhamnose, or polysaccharides such as raffinose and inulin.

[0206] Experimental Example 2. The effect of the composition of the present invention on improving insomnia.

[0207] Fifty-six patients with clinical insomnia, aged 25-60 years, were selected. The main clinical symptoms of insomnia included difficulty falling asleep, frequent awakenings during the night, early awakening with difficulty falling back asleep, and fatigue and poor concentration upon waking. Patients with long-term insomnia also experienced complications such as memory loss, mood swings, and weakened immunity.

[0208] With informed consent, the subjects were randomly divided into 8 groups of 7 people each, with 28 cases in the treatment group and 28 cases in the control group.

[0209] The treatment group was administered the probiotic composition formulations of Examples 1-4 of this invention, while the control group was administered the formulations of Comparative Examples 1-4. The oral formulations of Examples 1-4 and Comparative Examples 1-3 were administered once daily, 5g each time; Comparative Example 4 was administered 1 pill each time. Administration lasted 1-10 days (D1-D10).

[0210] The overall effect of the probiotic composition formulations of Examples 1-4 and the formulations of Comparative Examples 1-4 on improving the symptoms of insomnia is shown in Table 1.

[0211] Table 1.

[0212] As shown in Table 1, after taking the probiotic composition preparations of Examples 1-4 of this invention for 2-3 days, difficulty falling asleep improved, the number of times one woke up at night decreased, sleep time increased by 2-3 hours, and the mind was not groggy after waking up, with a good mental state, indicating a very good and rapid effect on improving insomnia; after taking it for 5-7 days, normal sleep could basically be restored. In Comparative Examples 1-3, due to a reduced number of live bacteria in the preparations or the lack of appropriate traditional Chinese medicine components or prebiotics, the effect of improving insomnia after taking the probiotic preparations was slower. The traditional Chinese medicine preparation in Comparative Example 4 also had a slow effect on improving insomnia. From the perspective of taste and compliance, the bitterness in Comparative Examples 1-3 was mainly caused by jujube seed, and the numbness was mainly caused by γ-aminobutyric acid (GABA). Patient compliance was poor, the preparations were slow to take effect, and the effect was not obvious. Comparative Example 2 attempted to improve the taste with mogrosides, but the effect on improving insomnia was not obvious. The probiotic complex of this application improves the taste by adding or subtracting ingredients and dosages of traditional Chinese medicine and prebiotics. The bitterness and numbing sensation are masked by adjusting the functional sugars. The preparation has no bitterness or strange taste, and the patient has good compliance. By adjusting the number of live bacteria, the preparation has a fast onset of action and a significant effect on improving insomnia.

[0213] Experimental Example 3. The effect of the composition of the present invention on improving insomnia caused by Helicobacter pylori.

[0214] Patient A, suffering from insomnia for over 10 years, experienced recurrent and severe insomnia. A C14 breath test revealed a DPM value of 1452 for Helicobacter pylori (reference range 0-100), and a serum total bilirubin level of 28.6 (reference range 1.7-17.1 μmol / L) in the five liver function tests.

[0215] The clinical diagnosis of Helicobacter pylori infection is quite serious. Gastroscopy results show chronic gastritis and gastric ulcers. The patient has clinical manifestations such as abdominal pain, bloating, nausea, and acid reflux, which affects sleep quality and causes insomnia.

[0216] Patient A took the probiotic composition solid beverage preparation prepared in Example 1, 5g once a day. The improvement of clinical symptoms is shown in Table 2.

[0217] Table 2.

[0218] As shown in Table 2, the patient had a relatively severe Helicobacter pylori infection, which caused insomnia. After taking the probiotic composition of this invention for 10 days, abdominal pain and bloating were reduced, nausea and acid reflux decreased, the frequency of insomnia decreased, and sleep duration increased; after one month of use, all the above symptoms disappeared, and sleep duration was 6-7 hours; after six months of use, the gastric ulcer healed, and sleep duration was 7-8 hours; after eight months of use, the DPM value of Helicobacter pylori detected by the C14 breath test was 0, indicating a complete cure for Helicobacter pylori infection and the insomnia it caused.

[0219] Experimental Example 4. The effect of the composition of the present invention on improving insomnia.

[0220] Clinical patient B experienced several months of stress and mental stimulation, leading to insomnia. After taking the preparation of Example 2 of this application for 1-2 days, 5g daily, sleep improved significantly. On the second day of taking the preparation, the patient woke up feeling clear-headed, in good mental condition, with reduced stress and a pleasant mood.

[0221] Clinical patient C suffered from insomnia for 3-5 years, with poor sleep quality, shallow sleep, and frequent awakenings. After taking the preparation of Example 3 of this application for 2-3 days, 5g daily, sleep improved significantly, sleep duration was prolonged, and deep sleep was well achieved. On the second day after taking the preparation, the patient woke up feeling clear-headed, energetic, and in a good mood.

[0222] Clinical patient Ding suffered from insomnia for 2-3 years, with sleep duration of 2-3 hours, and required the use of diazepam to aid sleep. After taking the preparation of Example 4 of this application for 3 days, 5g daily, sleep improved significantly, with sleep duration of 5-6 hours. The diazepam dosage was halved, and after one week, diazepam was discontinued. Sleep duration was 6-7 hours.

[0223] The above clinical results indicate that the composition of the present invention includes a probiotic complex and traditional Chinese medicine components that are both food and medicine, as well as prebiotics that promote the growth of probiotics and serve as their nutrient source. The probiotic complex contains more than 200 billion CFU / g of active probiotics. Combining probiotics with traditional Chinese medicine that is both food and medicine, and supplementing them with appropriate prebiotics, can treat or improve insomnia caused by emotional, stress, disease, etc., resulting in disharmony between the spleen and stomach, stagnation of liver qi, excessive heart fire, yin deficiency and excessive fire, and disharmony between the heart and kidneys. It has a rapid onset of action and a significant effect on improving insomnia.

[0224] The technical solutions of the present invention are not limited to the specific embodiments described above. Any technical modifications made in accordance with the technical solutions of the present invention fall within the protection scope of the present invention.

Claims

1. A composition, characterized in that, The composition comprises a probiotic complex and traditional Chinese medicine components, wherein the probiotic complex comprises or is composed of the following strains: *Lactobacillus plantarum*, *Bifidobacterium lactis*, *Lactobacillus crispatus*, *Lactobacillus acidophilus*, *Lactobacillus rhamnosus*, *Bifidobacterium bifidum*, *Lactobacillus reuteri*, *Lactobacillus casei*, *Lactobacillus bulgaricus*, *Lactobacillus fermentum*, *Lactobacillus gersoni*, *Lactococcus lactis*, *Bifidobacterium longum*, and *Lactobacillus salivarius*. The herbal components include or are composed of the following herbs: jujube seed, lily bulb, poria cocos, lotus seed, and mulberry.

2. The composition according to claim 1, characterized in that, The composition further includes a prebiotic selected from one or more of L-arabinose, β-glucan, inulin, erythritol, lactitol, vitamin C, and γ-aminobutyric acid.

3. The composition according to claim 2, characterized in that, The composition includes the probiotic complex, jujube seed, lily bulb, poria cocos, lotus seed, mulberry, L-arabinose, β-glucan, inulin, erythritol, lactitol, vitamin C, and γ-aminobutyric acid.

4. The composition according to any one of claims 1-3, characterized in that, The composition comprises the following components in parts by weight: 2-6 parts probiotic complex, 2-5 parts jujube seed, 2-7 parts lily bulb, 2-6.5 parts poria cocos, 2-8 parts lotus seed, 2-8 parts mulberry, 2-7 parts L-arabinose, 1-4 parts β-glucan, 5-10 parts inulin, 6-10 parts erythritol, 6-10 parts lactitol, 0.5-3 parts vitamin C, and 2-4.5 parts γ-aminobutyric acid.

5. The composition according to claim 4, characterized in that, The composition comprises the following components in parts by weight: 3-5 parts probiotic complex, 3-5 parts jujube seed, 2-5 parts lily bulb, 3-5 parts poria cocos, 2-5 parts lotus seed, 3-5 parts mulberry, 3.3-6.6 parts L-arabinose, 1-3 parts β-glucan, 6-8 parts inulin, 7-9 parts erythritol, 7-9 parts lactitol, 0.5-2 parts vitamin C, and 3-4.5 parts γ-aminobutyric acid; Preferably, the composition comprises the following components in parts by weight: 3 parts probiotic complex, 3 parts jujube seed, 3 parts lily bulb, 3 parts poria cocos, 2 parts lotus seed, 3 parts mulberry, 3.5 parts L-arabinose, 1.5 parts β-glucan, 7 parts inulin, 8 parts erythritol, 8 parts lactitol, 0.5 parts vitamin C, and 4.5 parts γ-aminobutyric acid.

6. The composition according to any one of claims 1-3, characterized in that, The probiotic complex contains the following live bacteria counts: *Lactobacillus plantarum* 850-950 billion CFU / g, *Bifidobacterium lactis* 50-150 billion CFU / g, *Lactobacillus curvatureii* 80-120 billion CFU / g, *Lactobacillus acidophilus* 100-180 billion CFU / g, *Lactobacillus rhamnosus* 50-150 billion CFU / g, *Bifidobacterium bifidum* 10-80 billion CFU / g, *Lactobacillus reuteri* 60-150 billion CFU / g, *Lactobacillus casei* 60-150 billion CFU / g, and *Lactobacillus bulgaricus*. Bacteria: 10-60 billion CFU / g, Lactobacillus fermentum: 60-100 billion CFU / g, Lactobacillus gasseri: 10-60 billion CFU / g, Lactococcus lactis: 10-60 billion CFU / g, Bifidobacterium longum: 20-80 billion CFU / g, Lactobacillus salivarius: 100-150 billion CFU / g, Bifidobacterium breve: 10-80 billion CFU / g, Streptococcus thermophilus: 50-100 billion CFU / g, Lactobacillus paracasei: 120-160 billion CFU / g, Lactobacillus helveticus: 50-100 billion CFU / g.

7. The composition according to claim 6, characterized in that, The probiotic complex contains the following live bacteria counts: *Lactobacillus plantarum* 90-93 billion CFU / g, *Bifidobacterium lactis* 10-13 billion CFU / g, *Lactobacillus curvatureii* 8-11 billion CFU / g, *Lactobacillus acidophilus* 13-16 billion CFU / g, *Lactobacillus rhamnosus* 10-13 billion CFU / g, *Bifidobacterium bifidum* 2-5 billion CFU / g, *Lactobacillus reuteri* 10-13 billion CFU / g, *Lactobacillus casei* 10-13 billion CFU / g, and *Bifidobacterium breve*. Lactobacillus lizarda 10-40 billion CFU / g, Lactobacillus fermentum 60-90 billion CFU / g, Lactobacillus gasseri 10-40 billion CFU / g, Lactococcus lactis 10-40 billion CFU / g, Bifidobacterium longum 30-60 billion CFU / g, Lactobacillus salivarius 100-130 billion CFU / g, Bifidobacterium breve 30-60 billion CFU / g, Streptococcus thermophilus 50-80 billion CFU / g, Lactobacillus paracasei 120-150 billion CFU / g, Lactobacillus helveticus 50-80 billion CFU / g; Preferably, the probiotic complex contains the following live bacteria counts: *Lactobacillus plantarum* 90 billion CFU / g, *Bifidobacterium lactis* 10 billion CFU / g, *Lactobacillus curvatureii* 8 billion CFU / g, *Lactobacillus acidophilus* 13 billion CFU / g, *Lactobacillus rhamnosus* 10 billion CFU / g, *Bifidobacterium bifidum* 2 billion CFU / g, *Lactobacillus reuteri* 10 billion CFU / g, *Lactobacillus casei* 10 billion CFU / g, *Lactobacillus bulgaricus* 1 billion CFU / g, *Lactobacillus fermentum* 6 billion CFU / g, *Lactobacillus gasseri* 1 billion CFU / g, *Lactococcus lactis* 1 billion CFU / g, *Bifidobacterium longum* 3 billion CFU / g, *Lactobacillus salivarius* 10 billion CFU / g, *Bifidobacterium breve* 3 billion CFU / g, *Streptococcus thermophilus* 5 billion CFU / g, *Lactobacillus paracasei* 12 billion CFU / g, and *Lactobacillus helveticus* 5 billion CFU / g.

8. A formulation, characterized in that, The formulation is made from the composition of any one of claims 1-7.

9. The formulation according to claim 8, characterized in that, The dosage form of the preparation is a solid beverage, a chewable tablet, or a lozenge.

10. Use of the composition of any one of claims 1-7 or the preparation of any one of claims 8-9 in the preparation of a medicament and / or food for treating or improving gastrointestinal inflammation, ulcers, Helicobacter pylori infection, constipation, insomnia.

Images