Application of probiotic composition in preparation of medicine for improving intestinal flora
By combining the probiotic composition of Sophora japonica flower powder, broccoli seed water extract, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis HN019, the problem of intestinal flora imbalance was solved, and the intestinal flora was restored and its health was optimized.
Patent Information
- Application Number
- CN202511057762.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-30
- Publication Date
- 2025-10-31
AI Technical Summary
There are no reports in the existing technology of using broccoli seed water extract, sophora japonica powder, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis to improve the intestinal flora, which leads to intestinal flora imbalance and causes a variety of diseases.
A probiotic composition is provided, comprising Sophora japonica powder, broccoli seed water extract, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis HN019, with optimized proportions and administration method to restore intestinal flora balance and diversity.
It significantly increases the abundance of beneficial bacteria, restores the balance of intestinal flora, improves the intestinal microecological imbalance index, optimizes the flora structure, and restores intestinal health.
Smart Images
Figure CN120860093A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical technology, and in particular relates to the application of a probiotic composition in the preparation of drugs that improve intestinal flora. Background Technology
[0002] The human gut is home to a large number of symbiotic microorganisms. These symbiotic microbial communities form the body's "second organ," not only cooperating with the host in the digestion and absorption of nutrients but also playing a vital role in maintaining human health. Under healthy conditions, the gut microbiota forms a balanced and stable micro-ecosystem in the gastrointestinal tract, maintaining bodily health. However, once the proportion of the microbiota changes significantly, the balance is disrupted, leading to dysbiosis, which can cause various diseases such as inflammatory bowel disease, irritable bowel syndrome, colorectal cancer, obesity, tumors, and depression.
[0003] Broccoli seed water extract is rich in glucoraphanin, which can be converted into sulforaphane after action by myrosinase or gut microbiota, activating the expression of transcription factor NRF2 to produce antioxidant effects. Sophora japonica flower powder is rich in quercetin, a flavonol compound; numerous in vitro and in vivo studies have shown that it plays a role in various tumor diseases by regulating biological processes such as oxidative stress and apoptosis. Lactobacillus rhamnosus GG is a third-generation probiotic, a safe and effective strain that promotes digestive health and maintains immune function. Bifidobacterium lactis HN019 is a strain isolated from yogurt. However, there are no reports on the combined use of broccoli seed water extract, Sophora japonica flower powder, Lactobacillus rhamnosus, and Bifidobacterium lactis to improve gut microbiota. Summary of the Invention
[0004] Therefore, the purpose of this invention is to provide an application of a probiotic composition in the preparation of a drug for improving intestinal flora.
[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:
[0006] This invention provides the application of a probiotic composition in the preparation of a drug to improve intestinal flora, the probiotic composition comprising Sophora japonica powder, broccoli seed water extract, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis.
[0007] Preferably, the *Lactobacillus rhamnosus* is *Lactobacillus rhamnosus* GG; and the *Bifidobacterium animalis* subsp. *mammus* is *Bifidobacterium animalis* subsp. *mammus* HN019.
[0008] Preferably, the probiotic composition can restore the balance and diversity of the intestinal flora, and significantly change the intestinal microecological imbalance index.
[0009] Preferably, the probiotic composition increases the abundance of beneficial bacteria.
[0010] Preferably, the beneficial bacteria are one or both of Bacteroides and Verrucous microbes.
[0011] Preferably, the mass ratio of the Sophora japonica powder to the broccoli seed water extract is 1:1; the effective viable count ratio of Lactobacillus rhamnosus to Bifidobacterium animalis subsp. lactis is (18-22):3; and the mass effective viable count ratio of the Sophora japonica powder to Lactobacillus rhamnosus is 1 mg: 5.5-6.5 billion CFU.
[0012] This invention provides the application of the above-mentioned probiotic composition in the preparation of a drug that promotes the growth of beneficial bacteria.
[0013] Preferably, the beneficial bacteria are one or both of Bacteroides and Verrucous microbes.
[0014] Preferably, the probiotic composition is the sole active ingredient of the drug.
[0015] The present invention provides a drug for improving gut microbiota, the drug comprising pharmaceutically acceptable excipients and the above-described probiotic composition.
[0016] Compared with the prior art, the present invention has the following beneficial effects:
[0017] This invention provides the application of a probiotic composition in the preparation of drugs for improving intestinal flora. Studies have found that the probiotic composition of this invention increases the abundance of beneficial bacteria, restores the balance and diversity of intestinal flora, and significantly alters the intestinal microecological imbalance index. Therefore, this probiotic composition has the effect of improving intestinal flora. This invention provides a new option for clinical drugs for improving intestinal flora. Attached Figure Description
[0018] Figure 1 To analyze the effects of probiotic compositions on mouse gut microbiota using 16S sequencing, A was principal component analysis (PCA) to assess the dispersion of microbiota among different groups; B was Venn diagram analysis to analyze differences in the detected phyla among different groups; C was the Kruskal-Wallis H statistical test to assess the impact of different groups on the diversity index; and D was a multi-group difference test to assess the phyla with significant differences in microbial composition among different groups. The x-axis represents the average relative abundance in different groups, and the y-axis represents the species name.
[0019] Figure 2In the middle section, bar chart A shows the effect of different groups on the enrichment of bacteria with relevant phenotypes, while bar B shows the results of intestinal microecological dysbiosis analysis for different groups. Detailed Implementation
[0020] This invention provides the use of a probiotic composition in the preparation of a drug for the prevention and / or treatment of pneumonia, wherein the probiotic composition comprises Sophora japonica powder, broccoli seed water extract, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis.
[0021] In this invention, as a preferred embodiment, the probiotic composition comprises Sophora japonica bud powder, broccoli seed water extract, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis. The Lactobacillus rhamnosus is preferably Lactobacillus rhamnosus GG; the Bifidobacterium animalis subsp. lactis is preferably Bifidobacterium animalis subsp. lactis HN019. This invention does not have specific limitations on the sources of the Sophora japonica bud powder, broccoli seed water extract, Lactobacillus rhamnosus GG, and Bifidobacterium animalis subsp. lactis HN019; commercially available products in the art can be used.
[0022] In this invention, the mass ratio of Sophora japonica bud powder to broccoli seed water extract is 1:1; the preferred ratio of effective viable bacteria count of Lactobacillus rhamnosus to Bifidobacterium animalis subsp. lactis is (18-22):3, more preferably (19-21):3, and even more preferably 20:3; the preferred ratio of effective viable bacteria count of Sophora japonica bud powder to Lactobacillus rhamnosus is 1 mg:5.5-6.5 billion CFU, more preferably 1 mg:5.7-6.3 billion CFU, and even more preferably 1 mg:6 billion CFU. This invention uses a probiotic composition with the above-mentioned content or effective viable bacteria count to increase the abundance of beneficial bacteria, wherein the beneficial bacteria are one or both of Bacteroides and Verrucous microbes. This probiotic composition can restore the balance and diversity of the intestinal flora, significantly changing the intestinal microecological imbalance index. This probiotic composition improves the abundance ratio (F / B) of Firmicutes and Bacteroides, optimizes the flora structure, and restores the balance of the intestinal flora.
[0023] Based on this, the present invention also provides the use of the above-mentioned probiotic composition in the preparation of a drug that promotes the growth of beneficial bacteria and / or inhibits harmful bacteria. The beneficial bacteria are one or both of Bacteroides and Verrucous microbes.
[0024] The present invention provides a drug for improving gut microbiota, the drug comprising pharmaceutically acceptable excipients and the above-described probiotic composition.
[0025] In this invention, the Sophora japonica powder, broccoli seed water extract, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis in the probiotic composition can be administered independently, or they can be mixed in a specific dosage and administered together. The administration methods include oral or injection.
[0026] In this invention, the drug comprises pharmaceutically acceptable excipients. The excipients include one or more of solvents, flavoring agents, excipients, antioxidants, and preservatives. The probiotic composition constitutes 60% to 90% of the drug by mass. The probiotic composition of this invention can be formulated into any conventional preparation using conventional methods, such as granules, tablets, lyophilized forms, capsules, syrups, or solutions.
[0027] In this invention, the drug can be used as the sole active ingredient of a probiotic composition to improve intestinal flora, or it can be combined with other active drugs that improve intestinal flora to achieve the same goal. Preferably, the probiotic composition is the sole active ingredient of the drug. When the probiotic composition of this invention improves intestinal flora, the effective dose range of the Sophora japonica powder and broccoli seed water extract is 4–7 mg / kg / day, the effective viable count of Lactobacillus rhamnosus is 60 billion CFU / kg / day, and the effective viable count of Bifidobacterium animalis subsp. lactis is 9 billion CFU / kg / day. The effective dose or effective viable count in this invention refers to the amount of the therapeutic agent used to treat, alleviate, or prevent the target disease or condition.
[0028] In this invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.
[0029] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0030] In the following examples, *Lactobacillus rhamnosus* GG was purchased from DSM-Firmenich, product code: 5018209, with a specification of 350 billion cfu / g.
[0031] Bifidobacterium animalis subsp. lactis HN019, purchased from Fonterra Cooperative Group Ltd., product code: 61018295, specification: 300 billion cfu / g.
[0032] The Sophora japonica powder was purchased from Idina Biotechnology (Shanghai) Co., Ltd., product number: 9047510A01.
[0033] Broccoli seed water extract was purchased from Ganzhou Huahan Biotechnology Co., Ltd., product number: 60202007.
[0034] MRS medium: tryptone 10 g / L, yeast extract 5 g / L, beef extract 10 g / L, glucose 20 g / L, sodium acetate 5 g / L, diammonium citrate 2 g / L, Tween 80 1 ml / L, magnesium sulfate 0.58 g / L, manganese sulfate 0.25 g / L, pH adjusted to approximately 6.2, sterilized at 115℃.
[0035] Example 1
[0036] (1) Pharmaceuticals and Pharmaceutical Preparation
[0037] S1. A probiotic composition for improving gut microbiota, wherein the probiotic composition comprises the following components:
[0038] Sophora japonica powder, broccoli seed water extract, Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis HN019.
[0039] Among them, 1 mg / mL Sophora japonica powder solution and 1 mg / mL broccoli seed water extract solution were prepared by using physiological saline.
[0040] Lactobacillus rhamnosus GG: Incubate Lactobacillus rhamnosus GG on MRS medium at 37°C for 20 hours, centrifuge at 7000 rpm for 30 minutes, and collect the precipitate to obtain Lactobacillus rhamnosus GG.
[0041] Bifidobacterium animalis subsp. lactis HN019: Bifidobacterium animalis subsp. lactis HN019 was cultured in MRS medium at 37°C for 20 hours, centrifuged at 7000 rpm for 30 min, and the precipitate was collected to obtain Bifidobacterium animalis subsp. lactis HN019.
[0042] S2, Dexamethasone solution: Prepared with physiological saline to a final concentration of 500 μg / mL.
[0043] S3. Preparation of lipopolysaccharide (LPS) solution: A lipopolysaccharide solution with a final concentration of 2 mg / mL was prepared using physiological saline.
[0044] (2) Experimental grouping and drug administration
[0045] C57BL / 6 mice (female, 4 weeks old, 18-22g) were divided into four groups of 6-8 mice each: blank group (Vehicle), model group (LPS), LPS+dexamethasone group (LPS+Dexa), and LPS+probiotic combination group (LPS+Combined).
[0046] In this study, the blank control group and the model group were administered physiological saline to each C57BL / 6 mouse by gavage once daily for 3 consecutive days; the LPS + dexamethasone group was administered dexamethasone solution at a dose of 5 mg / kg to each C57BL / 6 mouse by gavage once daily for 3 consecutive days; the LPS + probiotic combination group was administered the above-mentioned Sophora japonica powder solution at a dose of 10 mg / kg, the above-mentioned broccoli seed water extract solution at a dose of 10 mg / kg, and the above-mentioned probiotic combination solution at a dose of 60 billion CFU / kg to each C57BL / 6 mouse. Mice were administered a probiotic composition by gavage, consisting of *Lactobacillus rhamnosus* GG and *Bifidobacterium lactis* subsp. *HN019* at a concentration of 9 billion CFU / kg, once daily for 3 consecutive days. One hour after the last administration, except for the control group, each mouse in the model group, LPS+dexamethasone group, and LPS+probiotic composition group was treated with a single intranasal instillation of 5 mg / kg of the above-mentioned LPS solution for 12 hours. Feces from each group were collected, and the effects of different treatments on the changes in the intestinal flora of mice were analyzed by 16S sequencing.
[0047] Figure 1 The results of the A study showed that PLS-DA analysis revealed significant differences in gut microbiota between the LPS model group and the control group. Furthermore, compared to the control group or the LPS group, both the LPS+dexamethasone group and the LPS+probiotic combination group exhibited a certain degree of dispersion, suggesting that all treatments interfered with the distribution of the gut microbiota to some extent.
[0048] Figure 1 Classification analysis of B bacteria showed that only 15.86% of the phyla were detected under all treatment conditions, suggesting that the distribution of gut microbiota in mice in the LPS group, LPS+dexamethasone group and LPS+probiotic combination group was significantly different.
[0049] Figure 1 Box plots of the gut microbiota diversity index (Ace) in group C showed that LPS significantly increased the Ace index (p<0.05), while the Ace indexes of the LPS+dexamethasone group and the LPS+probiotic combination group decreased, suggesting that dexamethasone and probiotic combination treatment could partially reverse the gut microbiota dysbiosis caused by LPS, thereby restoring gut microbiota diversity. Specifically, compared with the control group, the relative abundance of Firmicutes was reduced and the relative abundance of Bacteroidetes was increased in the LPS group, and the abundance ratio of Firmicutes to Bacteroidetes (F / B value) was reduced. The imbalance of F / B value is closely related to chronic inflammation. In the LPS+dexamethasone and LPS+probiotic combination groups, the supplementation of prebiotics and probiotics optimized the gut microbiota structure, improved the F / B value, and restored the balance of gut microbiota.
[0050] Furthermore, compared with the control group and the LPS group, the LPS + dexamethasone group and the LPS + probiotic combination group increased the relative abundance of Verrucous microbes. Verrucous microbes have anti-inflammatory properties, and their abundance is closely related to gut health, suggesting that the LPS + dexamethasone group and the LPS + probiotic combination group can partially reverse the LPS-induced dysbiosis (see...). Figure 1 (D in the middle).
[0051] Figure 1 The results showed that the combination of LPS+dexamethasone and LPS+probiotics could partially reverse the LPS-induced dysbiosis.
[0052] Figure 2 Disease-related phenotype predictions suggest that, compared with the LPS group, the proportion of disease-related flora (especially pathogenic potentially) was reduced in the LPS+probiotic combination group, initially suggesting that the probiotic combination has a potential role in regulating the gut microbiota.
[0053] Specifically, the control group had a higher proportion of Firmicutes, while the relative abundance of Firmicutes decreased after LPS treatment. In the LPS + probiotic combination group, the proportions of Verrucous and Bacteroidetes were higher, suggesting that the probiotic combination may improve the gut microbiota by increasing the relative abundance of beneficial bacteria. Compared to the control group, the LPS group showed an increased relative abundance of anaerobic, facultative anaerobic, pathogenic, and biofilm-forming bacteria, and a decreased relative abundance of aerobic bacteria containing motile elements. Biofilm-forming bacteria exhibit strong resistance to antibiotics and host immune defense mechanisms, potentially affecting the efficacy of antibiotics. In the LPS + probiotic combination group, the relative abundance of anaerobic and Gram-negative bacteria increased (see...). Figure 2 A in the middle.
[0054] Figure 2 Analysis of the gut microbiota dysbiosis index in group B showed that, compared with the LPS group, treatment with dexamethasone and probiotic combination significantly altered the gut microbiota dysbiosis index, suggesting that the probiotic combination has a potential regulatory effect on gut microbiota.
[0055] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. The use of a probiotic composition in the preparation of drugs for improving intestinal flora, characterized in that, The probiotic composition includes Sophora japonica powder, broccoli seed water extract, Lactobacillus rhamnosus, and Bifidobacterium animalis subsp. lactis.
2. The application according to claim 1, characterized in that, The Lactobacillus rhamnosus is Lactobacillus rhamnosus GG; the Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis HN019.
3. The application according to claim 1, characterized in that, The probiotic composition can restore the balance and diversity of the intestinal flora, and significantly change the intestinal microecological imbalance index.
4. The application according to claim 1, characterized in that, The probiotic composition increases the abundance of beneficial bacteria.
5. The application according to claim 4, characterized in that, The beneficial bacteria are one or both of Bacteroides and Verrucous microbes.
6. The application according to claim 1, characterized in that, The mass ratio of Sophora japonica bud powder to broccoli seed water extract is 1:1; the effective viable count ratio of Lactobacillus rhamnosus to Bifidobacterium animalis subsp. lactis is (18-22):3; and the mass effective viable count ratio of Sophora japonica bud powder to Lactobacillus rhamnosus is 1 mg: 5.5-6.5 billion CFU.
7. The use of the probiotic composition according to any one of claims 1 to 6 in the preparation of a drug that promotes the growth of beneficial bacteria.
8. The application according to claim 7, characterized in that, The beneficial bacteria are one or both of Bacteroides and Verrucous microbes.
9. The application according to claim 1 or 7, characterized in that, The probiotic composition is the sole active ingredient of the drug.
10. A drug for improving intestinal flora, characterized in that, The drug comprises pharmaceutically acceptable excipients and the probiotic composition of any one of claims 1 to 9.