An antibacterial composition and its preparation method and application

Abstract

The present invention discloses an antibacterial composition comprising the following components by weight: 4-6 parts vitamin C, 1-2 parts vitamin E, 1-2 parts angelica polysaccharide, and 1-2 parts ligustrum lucidum polysaccharide. The present invention also discloses a preparation method of the antibacterial composition and its use in preparing a medicament for inhibiting bacteria and / or fungi or treating inflammation.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to an antibacterial composition, a preparation method and an application thereof. Background Art

[0002] Infection is a pathological condition in which pathogens (such as bacteria, viruses, and fungi) invade the body, triggering an immune system response. During infection, inflammatory factors in peripheral blood act as immune regulatory molecules and play a key role in the host's defense response. The dynamic changes in inflammatory factors can reflect the intensity and characteristics of the body's response to infection, making peripheral blood testing an important tool for assessing infectious diseases. In certain pathogen infections, excessive release of inflammatory factors may lead to an "inflammatory storm," which in turn triggers systemic inflammatory response syndrome (SIRS) and multiple organ failure (MODS). In patients with sepsis caused by bacterial infection, levels of TNF-α, IL-6, and other proteins in the peripheral blood are significantly elevated, closely related to the severity of the disease and prognosis. Novel coronavirus infection can trigger excessive release of proinflammatory factors (such as IL-6 and interferon-γ), leading to lung damage and acute respiratory distress syndrome (ARDS). Fungal infections, such as Candida albicans infection, trigger a systemic inflammatory response by activating the release of proinflammatory factors TNF-α and IL-1β.

[0003] Bacterial infections have become a major challenge to global public health. In 2019, bacterial infections caused approximately 7.7 million deaths, accounting for 13.6% of all global deaths and becoming the second leading cause of death worldwide, after ischemic heart disease. Staphylococcus aureus, Escherichia coli, Streptococcus pneumoniae, Klebsiella pneumoniae, and Pseudomonas aeruginosa were the leading pathogens responsible for these deaths. These bacteria cause infections such as lower respiratory tract infections, bloodstream infections, and intra-abdominal infections. Antimicrobial resistance (AMR) is a growing problem, making bacterial infections more difficult to treat. A 2022 World Health Organization report revealed high levels of antibiotic resistance in bacteria causing fatal bloodstream infections, and increasing resistance to treatment for several common community-acquired bacteria. Bacterial infections pose a significant threat to global health, particularly in the context of rising antimicrobial resistance. Strengthening infection prevention, optimizing antibiotic use, improving microbial detection capabilities, and developing new antimicrobial drugs and vaccines are crucial to reducing the burden of bacterial infections.

[0004] However, current antimicrobial preparations remain suboptimal in efficacy or stability. Many traditional antimicrobial preparations have a single source of active ingredients and a relatively simple formulation structure. Traditional antimicrobial regimens often focus on using a single chemical drug or specific Chinese herbal extract, overlooking the potential synergistic effects between nutrients, antioxidants, and the individual Chinese herbal components. This lacks multifaceted synergy in the in vivo function of the active ingredients, resulting in insufficient comprehensive regulatory capacity under physiological conditions and, consequently, less stable or significant clinical efficacy than anticipated. Furthermore, there may be side effects and safety concerns. Some commonly used antimicrobial drugs are prone to adverse reactions such as gastrointestinal irritation and increased liver and kidney burden with long-term use. This is due to the potential irritation or damage to corresponding organs during metabolism and excretion by some chemical drugs. Furthermore, existing antimicrobial preparations have a relatively simple mechanism of action and lack overall regulatory capacity. Traditional antimicrobial drugs often target specific pathways in the antimicrobial response, but the infection process is complex and not limited to a single pathway. The lack of a comprehensive, multi-target, multi-pathway regulatory approach may result in the inability of these drugs to provide comprehensive and lasting relief of infection in practice. Summary of the Invention

[0005] One object of the present invention is to provide an antibacterial composition with a sustained, mild and efficient antibacterial effect in response to the above technical problems.

[0006] Another object of the present invention is to provide a method for preparing the antibacterial composition.

[0007] Another object of the present invention is to provide applications of the antibacterial composition.

[0008] In order to achieve the above object of the invention, the present invention provides an antibacterial composition, which comprises the following components by weight: 4-6 parts of vitamin C, 1-2 parts of vitamin E, 1-2 parts of angelica polysaccharide, and 1-2 parts of Ligustrum lucidum polysaccharide.

[0009] As a preferred embodiment, the weight ratio of vitamin C, vitamin E, angelica polysaccharide and Ligustrum lucidum polysaccharide is 5:1:1:1.

[0010] As a preferred embodiment, the antibacterial composition further comprises a solvent. Preferably, the solvent is physiological saline, glucose solution or other solvents suitable for injection, such as water.

[0011] As a preferred embodiment, the antibacterial composition further comprises a stabilizer. Further preferably, the stabilizer is an antioxidant. More preferably, the antioxidant is selected from one or more of tropol, panthenol, and butylated hydroxyanisole (BHT).

[0012] On the other hand, the present invention also provides a method for preparing the antibacterial composition, which comprises the following steps:

[0013] (1) Add angelica polysaccharide and glossy privet polysaccharide into water and stir to dissolve;

[0014] (2) adding vitamin C powder and stirring until completely dissolved to obtain a mixed solution;

[0015] (3) Mixing vitamin E emulsifier and vegetable oil, stirring evenly, then slowly adding it to the above mixed solution, stirring homogenously, and making it evenly dispersed to form a stable dispersion;

[0016] (4) Drying, crushing, and sieving the above dispersion to obtain the antibacterial composition.

[0017] On the other hand, the present invention also provides use of the antibacterial composition in the preparation of a medicament for inhibiting bacteria and / or fungi.

[0018] As a preferred embodiment, the bacteria are Gram-negative bacteria or Gram-positive bacteria.

[0019] As a preferred embodiment, the bacteria are one or more of the following: Gram-negative bacteria including Escherichia coli, Shigella flexneri, Shigella sonnei, Klebsiella pneumoniae, Gram-positive bacteria including Staphylococcus aureus, and fungi including Candida albicans.

[0020] On the other hand, the present invention also provides the use of the antibacterial composition in preparing a drug for treating inflammation.

[0021] Preferably, the antibacterial composition of the present invention can be prepared into various known dosage forms, including but not limited to solid dosage forms, semisolid dosage forms, liquid dosage forms, gas and aerosol dosage forms, implant dosage forms and special dosage forms.

[0022] Preferably, the antibacterial composition of the present invention can be administered in a variety of different ways, including but not limited to oral administration, injection, transdermal administration, inhalation, topical administration, rectal administration, sublingual or buccal administration, implantation and special route administration.

[0023] Preferably, the antibacterial composition of the present invention can be administered at a dose of about 8-80 mg / kg body weight / day.

[0024] As a preferred embodiment, the inflammation includes inflammation caused by immune response, cytokine activation or oxidative damage, such as rheumatoid arthritis, arteriosclerosis, hepatitis and skin diseases.

[0025] Vitamin C is a water-soluble antioxidant that can directly scavenge ROS and inhibit the release of inflammatory factors. Vitamin C supplementation can reduce the levels of inflammatory markers C-reactive protein (CRP) and IL-6, and improve a variety of chronic inflammatory-related diseases. Liposome-encapsulated vitamin C regulates the inflammatory factors IL-6, MCP-1, and MIP-1α, and has been found to significantly improve immune function. Vitamin C plays an important role in reducing proinflammatory factors and controlling inflammation. Vitamin C (ascorbic acid) has certain effects in immunomodulation and antibacterial properties.

[0026] Vitamin E is a fat-soluble antioxidant that protects cell membranes from oxidative damage by preventing lipid peroxidation. Vitamin E has shown a significant effect in reducing TNF-α and IL-6 levels, especially under conditions of oxidative stress. Studies on the effects of vitamin E on proinflammatory factors in patients with schizophrenia have revealed a significant role in anti-oxidation and inflammation regulation. Vitamin E has potential value in reducing postprandial inflammatory responses, especially its ability to regulate inflammatory factors. Members of the vitamin E family have a protective effect against radiation-induced inflammation, effectively mitigating radiation damage by regulating the expression of inflammatory factors. Furthermore, vitamin E can enhance the bactericidal effect of antibiotics by inhibiting the binding of bacterial lipocalin (such as BcnA produced by Burkholderia cenocepacia) to antibiotics.

[0027] Angelica polysaccharides are an active ingredient extracted from the traditional Chinese medicine Angelica sinensis, and they have a wide range of functions. Angelica polysaccharides have shown a certain inhibitory effect on various tumor cells in vitro, and its mechanism may be related to inducing cell apoptosis and inhibiting tumor cell proliferation. Angelica polysaccharides have a certain hypoglycemic effect in diabetic rats, and its mechanism may be achieved by improving pancreatic islet function and regulating insulin sensitivity. As a multifunctional drug, Angelica polysaccharides have been shown to have a wide range of pharmacological effects, including antioxidant, immunomodulatory, and hematopoietic promotion. Angelica polysaccharides can significantly promote the proliferation and differentiation of hematopoietic stem cells and enhance the hematopoietic function of the bone marrow.

[0028] The chemical components of Fructus ligustri lucidi polysaccharides include polysaccharides, phenolic acid compounds, and triterpenoids. Research has shown their multiple anti-aging, immunomodulatory, and anti-tumor effects, with specific mechanisms including free radical scavenging, immune cell enhancement, and regulation of tumor-related factors. Fructus ligustri lucidi polysaccharides have significant anti-aging effects, inhibiting the degeneration of immune organs, enhancing immune function, and scavenging free radicals. Fructus ligustri lucidi polysaccharides have inhibitory effects on certain tumor cells, possibly through enhancing immune function and inhibiting tumor cell proliferation. Fructus ligustri lucidi polysaccharides can enhance immunity by regulating T cell activity and enhancing macrophage function. The synergistic use of Fructus ligustri lucidi polysaccharides and Cuscuta australis polysaccharides significantly improves anti-aging effects, possibly through enhanced antioxidant capacity. Fructus ligustri lucidi polysaccharides can also increase the antigenicity of lymphoma cell membranes, thereby enhancing the immune response to tumors.

[0029] By rationally combining vitamin C, vitamin E with angelica polysaccharides and glossy privet polysaccharides in a specific ratio, and utilizing the different bactericidal and anti-inflammatory mechanisms of the four to form a synergistic effect, the composition of the present invention achieves a composite antibacterial effect, improves the overall antibacterial and anti-inflammatory efficacy and stability, thereby obtaining better treatment and preventive effects in clinical applications. Experiments have shown that the composition of the present invention can significantly reduce the concentration of peripheral blood inflammatory factors (such as TNF-α, IL-6) and promote peripheral blood sterilization. This multi-pathway synergistic effect can provide a more comprehensive bactericidal and anti-inflammatory effect compared to traditional single drug treatment.

[0030] Compared with the prior art, the present invention improves the stability and sustainability of the overall antibacterial effect. By utilizing the synergistic effect of multiple components, the comprehensive regulatory ability of the effective ingredients in the body is improved, so that the final preparation is improved in terms of stability and efficacy. In addition, the present invention also reduces the risk of potential adverse reactions and side effects. When selecting and matching ingredients, the biosafety of Chinese medicine monomer components and vitamins is taken into account, in order to reduce the adverse stimulation and damage to the gastrointestinal tract, liver and kidneys that may be caused by traditional chemical antibacterial drugs. Furthermore, the present invention further enriches the antibacterial mechanism of action and enhances the overall conditioning ability. With the help of the multi-link intervention effect of vitamins and Chinese medicine monomer components, it is expected to appropriately balance and regulate multiple signal pathways in the antibacterial process, so as to obtain a more comprehensive, lasting and mild antibacterial effect in clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The colony growth of E. coli is shown.

[0032] Figure 2 The relative number of viable bacteria for the E. coli culture experiment is shown.

[0033] Figure 3The colony growth of Shigella flexneri is shown.

[0034] Figure 4 The relative number of viable bacteria from the Shigella flexneri culture experiment is shown.

[0035] Figure 5 Shown are the IL-6 cytokine concentrations in plasma after the addition of E. coli.

[0036] Figure 6 Shown are the IL-6 cytokine concentrations in plasma after the addition of Shigella flexneri . DETAILED DESCRIPTION

[0037] The present invention will be further described below with reference to specific examples. It should be understood that the following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0038] Vitamin C is a commercial raw material purchased from Shanghai Biyuntian Biotechnology Co., Ltd. with the product number ST1434.

[0039] Vitamin E is a commercial raw material purchased from Shanghai Biyuntian Biotechnology Co., Ltd., product number S0079.

[0040] Angelica polysaccharide is a commercial raw material purchased from Shanghai Yuanye Biotechnology Co., Ltd., product number S27815.

[0041] Ligustrum lucidum polysaccharide is a commercial raw material purchased from Shanghai Ronghe Pharmaceutical Technology Development Co., Ltd. with the product number TDT044.

[0042] Unless otherwise specified, the reagents, instruments, etc. used in the examples of the present invention are well known to those skilled in the art and can be purchased through commercial channels.

[0043] Example

[0044] 1. Preparation of antibacterial composition (taking the preparation of solid composition as an example)

[0045] (1) Add 2 g of Angelica sinensis polysaccharide and 2 g of Ligustrum lucidum polysaccharide to 200 mL of purified water and stir to dissolve evenly. If necessary, gently heat in a 40-60°C water bath to accelerate the dissolution and dispersion of the polysaccharide until a uniform polysaccharide solution is obtained. The pH and temperature of the polysaccharide solution can be appropriately controlled according to the characteristics of the polysaccharide and the requirements of the subsequent steps, and are usually in the neutral or slightly acidic range, such as pH 6-7.

[0046] (2) Slowly add 10 g of vitamin C (L-ascorbic acid) to the polysaccharide solution and continue stirring until the vitamin C is completely dissolved to obtain a clear, uniform polysaccharide-vitamin C mixed solution. Vitamin C is water-soluble and generally dissolves easily.

[0047] (3) 2 g of vitamin E is first mixed with 1 g of a food-grade emulsifier (such as lecithin) and 1 mL of vegetable oil, and a uniform emulsion is formed by high shear stirring or ultrasonic dispersion. Then, the mixture is slowly added dropwise to the polysaccharide-vitamin C mixed solution while stirring to form a stably dispersed emulsion system of vitamin E. Other similar methods can also be used to make the vitamin E in a relatively uniform and stable dispersion state in the system. The emulsion system can be homogenized or stirred to ensure that the oil phase (vitamin E) in the system is refined and stably distributed in the aqueous phase.

[0048] (4) The uniform dispersion obtained above is spray-dried or vacuum-freeze-dried to obtain a solid powder composition containing vitamin C, vitamin E, angelica polysaccharide and Ligustrum lucidum polysaccharide.

[0049] (5) The dried solid powder composition is crushed and sieved to obtain a final product, which can be further capsule filled, tablet pressed or prepared into a granular preparation according to the final purpose.

[0050] 2. Bacterial Culture

[0051] In order to study and evaluate the in vitro bacterial growth inhibition efficacy of the antibacterial composition of the present invention, a bacterial culture experiment was performed.

[0052] Experimental groups: There were four groups in total. (1) Blank control group: normal saline; (2) Multivitamin group: a complex of vitamin C and vitamin E, with a weight ratio of 5:1; (3) Chinese herbal polysaccharide group: a complex of angelica polysaccharide and Ligustrum lucidum polysaccharide, with a weight ratio of 1:1; (4) Combined medication group: a combination of vitamin C, vitamin E, angelica polysaccharide, and Ligustrum lucidum polysaccharide prepared above, with a weight ratio of 5:1:1:1 among these three groups.

[0053] Escherichia coli (Beina Biotech, Catalog No. BNCC336902) and Shigella flexneri (Beina Biotech, Catalog No. BNCC232380) were cultured in nutrient broth (LB) liquid medium (Shanghai Beyotime Biotechnology Co., Ltd., Catalog No. ST163) at 37°C overnight. The bacterial concentration was then adjusted to a McFarland concentration of 1.0 with physiological saline to obtain a bacterial suspension.

[0054] 3. Bacterial stimulation of peripheral blood experiments

[0055] In order to study and evaluate the antibacterial and anti-inflammatory effects of the antibacterial composition of the present invention on peripheral blood, a bacterial stimulation peripheral blood experiment was performed.

[0056] Peripheral blood was collected from six healthy adult subjects using glass tubes containing lithium heparin anticoagulant. 500 μL of anticoagulated whole blood was transferred to sterile, additive-free glass tubes and divided into four groups. The aforementioned compositions were added to each group (blank control group, the same volume of normal saline as the anticoagulated whole blood; multivitamin group, 60 μg / ml; traditional Chinese medicine polysaccharide group, 20 μg / ml; combination drug group, 80 μg / ml) and thoroughly mixed.

[0057] Add 50 μl of bacterial suspension (Escherichia coli or Shigella flexneri) to the glass tube containing the composition and whole blood, mix thoroughly, and incubate at 37°C for 2 hours.

[0058] Fifty microliters of whole blood containing the above composition and bacterial suspension was taken and spread onto a 10-cm plate containing nutrient broth (LB) solid culture medium (Shanghai Biyuntian Biotechnology Co., Ltd., catalog number ST163), cultured overnight at 37°C, and the number of bacterial monoclonal colonies on the plate was counted to estimate the relative number of live bacteria in the whole blood.

[0059] The experimental results are as follows Figures 1 to 4 shown. Figure 1 The colony growth of E. coli is shown. Figure 2 The relative number of viable bacteria for the E. coli culture experiment is shown. Figure 3 The colony growth of Shigella flexneri is shown. Figure 4 The relative number of viable bacteria from the Shigella flexneri culture experiment is shown. Figures 1 to 4 The results showed that the multivitamin group significantly promoted peripheral blood bactericidal activity, the Chinese herbal polysaccharide group had no significant effect, and the combination group significantly promoted peripheral blood bactericidal activity. Compared with the multivitamin group, the combination group had a more significant effect in promoting peripheral blood bactericidal activity. This indicates that the antibacterial composition of the present invention has a good bactericidal effect, particularly against Escherichia coli and Shigella flexneri. In addition, the antibacterial composition of the present invention can also significantly inhibit Gram-negative bacteria such as Shigella sonnei and Klebsiella pneumoniae, Gram-positive bacteria such as Staphylococcus aureus, and fungi such as Candida albicans, and the effect is better than that of the multivitamin group and the Chinese herbal polysaccharide group.

[0060] The concentration of IL-6 cytokine in plasma was detected by electrochemiluminescence method on Roche cobas e 601 electrochemiluminescence immunoassay.

[0061] The results of IL-6 cytokine concentration test were as follows Figure 5 and Figure 6 As shown. Among them, Figure 5 Shown are the IL-6 cytokine concentrations in plasma after the addition of E. coli. Figure 6The results show the plasma IL-6 cytokine concentrations after the addition of Shigella flexneri. The results indicate that the multivitamin group significantly inhibited bacterial-induced secretion of the inflammatory cytokine IL-6 in peripheral blood, while the Chinese herbal polysaccharide group had no significant effect. The combined drug group significantly inhibited bacterial-induced secretion of the inflammatory cytokine IL-6 in peripheral blood. Compared to the multivitamin group, the combined drug group significantly inhibited bacterial-induced secretion of the inflammatory cytokine IL-6 in peripheral blood.

[0062] It can be seen that the antibacterial composition of the present invention has significantly better anti-inflammatory effect.

Claims

1. An antibacterial composition, characterized in that The antibacterial composition is prepared from the following components by weight: 4-6 parts of vitamin C, 1-2 parts of vitamin E, 1-2 parts of angelica polysaccharide, and 1-2 parts of ligustrum lucidum polysaccharide; the antibacterial composition inhibits Escherichia coli or Shigella flexneri.

2. The antibacterial composition according to claim 1, characterized in that The weight ratio of vitamin C, vitamin E, angelica polysaccharide and ligustrum lucidum polysaccharide is 5:1:1:

1.

3. The antibacterial composition according to claim 1, characterized in that The antimicrobial composition further comprises a solvent.

4. The antibacterial composition according to claim 3, characterized in that The solvent is water.

5. The antibacterial composition according to claim 1, characterized in that The antimicrobial composition further comprises a stabilizer.

6. The antibacterial composition according to claim 5, characterized in that The stabilizer is an antioxidant.

7. The method for preparing the antibacterial composition according to any one of claims 1 to 6, characterized in that The following steps are involved: (1) Add Angelica sinensis polysaccharide and Ligustrum lucidum polysaccharide into water and stir to dissolve; (2) Add vitamin C powder and stir until completely dissolved to obtain a mixed solution; (3) Mix the vitamin E emulsifier and vegetable oil, stir evenly, then slowly add it to the above mixed solution, stir homogenously, and disperse it evenly to form a stable dispersion; (4) Drying, crushing, and sieving the above dispersion to obtain the antibacterial composition.

8. Use of the antibacterial composition according to any one of claims 1 to 6 in the preparation of a medicament for inhibiting bacteria; the bacteria are Escherichia coli or Shigella flexneri.

9. Use of the antibacterial composition according to any one of claims 1 to 6 in the preparation of a medicament for treating inflammation caused by Escherichia coli or Shigella flexneri.

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