Antibacterial composition as well as preparation method and application thereof

Abstract

The invention discloses an antibacterial composition which comprises the following components in parts 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 glossy privet fruit polysaccharide. The invention also discloses a preparation method of the antibacterial composition and application of the antibacterial composition in preparation of medicines for inhibiting bacteria and / or fungi or treating inflammation.

Description

Technical Field

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

[0002] Infection is a pathological state in which pathogens (such as bacteria, viruses, fungi, etc.) invade the body and trigger the immune system to respond. During the infection process, inflammatory factors in peripheral blood, as immune regulatory molecules, play a key role in the host defense response. The dynamic changes of inflammatory factors can reflect the intensity and characteristics of the body's response to infection, so peripheral blood testing has become an important means of evaluating infectious diseases. In some pathogen infections, excessive release of inflammatory factors may lead to an "inflammatory storm", which in turn leads to systemic inflammatory response syndrome (SIRS) and multiple organ failure (MODS). In patients with sepsis caused by bacterial infection, the levels of TNF-α, IL-6, etc. in peripheral blood are significantly increased, which is closely related to the severity of the disease and prognosis. New coronavirus infection can trigger excessive release of proinflammatory factors (such as IL-6 and interferon-γ), leading to lung injury and acute respiratory distress syndrome (ARDS). Fungal infections, such as Candida albicans infection, trigger systemic inflammatory responses 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 the total global death toll, second only to ischemic heart disease, becoming the world's second leading cause of death. Among them, Staphylococcus aureus, Escherichia coli, Streptococcus pneumoniae, Klebsiella pneumoniae and Pseudomonas aeruginosa are the main pathogens causing death. The types of infections caused by these bacteria include lower respiratory tract infections, bloodstream infections and intra-abdominal infections. The growing problem of antimicrobial resistance (AMR) has made bacterial infections more difficult to treat. In 2022, the World Health Organization reported that the level of bacterial resistance that causes fatal bloodstream infections is high, and several bacteria that are common infections in the community are also becoming more resistant to treatment. Bacterial infections pose a major threat to global health, especially 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 essential to reducing the burden of bacterial infections.

[0004] However, the efficacy of current antibacterial preparations is still not ideal or the stability is insufficient. The effective ingredients of many traditional antibacterial preparations are from a single source, and the formula structure is relatively simple. Traditional antibacterial schemes often focus on the use of a certain chemical drug or a specific Chinese medicine extract alone, ignoring the potential synergistic effect between nutrients, antioxidants and Chinese medicine monomers. The process of active ingredients taking effect in the body lacks multi-level synergistic effects, which makes the comprehensive regulation ability of the formula under physiological conditions insufficient, resulting in clinical efficacy that is not as stable or significant as expected. In addition, there may be certain side effects and safety hazards. Some commonly used antibacterial drugs are prone to adverse reactions such as gastrointestinal irritation and increased liver and kidney burden during long-term use. This is because some chemical drugs have potential stimulating or damaging effects on the corresponding organs during metabolism and excretion. Furthermore, the mechanism of action of existing antibacterial preparations is relatively single, and the overall regulation ability is lacking. Traditional antibacterial drugs often focus on specific links in the antibacterial response, but the infection process is more complicated and is not limited to a single pathway. The lack of a comprehensive regulation scheme with multiple targets and multiple pathways may result in the inability of drugs to fully and permanently relieve infections in actual use. 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 view of the above technical problems to be solved.

[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 application of the antibacterial composition.

[0008] In order to achieve the above invention objectives, 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) adding angelica polysaccharide and Ligustrum lucidum polysaccharide into water and stirring 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, and then slowly adding the vitamin E to the mixed solution, stirring and homogenizing to make 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 the use of the antibacterial composition in the preparation of a drug 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 a variety of 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 dosage of about 8-80 mg / kg body weight / day, etc.

[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 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 a certain role 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 oxidative stress. The effect of vitamin E on proinflammatory factors in patients with schizophrenia has been found to have 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 on radiation-induced inflammation, and they effectively reduce radiation damage by regulating the expression of inflammatory factors. In addition, 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, which has a wide range of functions. Angelica polysaccharides show 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 bone marrow.

[0028] The chemical components of Fructus ligustri lucidi polysaccharides include polysaccharides, phenolic acid compounds and triterpenoid compounds. Its research covers multiple effects in anti-aging, immunomodulation and anti-tumor. The specific mechanisms include free radical scavenging, enhancing immune cell function and regulating tumor-related factors. Fructus ligustri lucidi polysaccharides have significant anti-aging effects, can inhibit the degeneration of immune organs, enhance the body's immune function, and scavenge free radicals. Fructus ligustri lucidi polysaccharides have an inhibitory effect on certain tumor cells, and its mechanism may involve enhancing immune function and inhibiting tumor cell proliferation. Fructus ligustri lucidi polysaccharides can improve immunity by regulating T cell activity and enhancing macrophage function. The synergistic use of Fructus ligustri lucidi polysaccharides and Cuscuta australis polysaccharides can significantly improve the anti-aging effect, and its mechanism may be related to enhancing antioxidant capacity. Fructus ligustri lucidi polysaccharides can increase the antigenicity of lymphoma cell membranes, thereby enhancing the body's immune response to tumors.

[0029] By rationally combining vitamin C, vitamin E with angelica polysaccharide and Ligustrum lucidum polysaccharide 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, and thus obtains better treatment and prevention 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. The synergistic effect of multiple components is utilized to improve the comprehensive regulatory ability of the effective ingredients in the body, 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 3Colony growth of Shigella flexneri is shown.

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

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

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

[0037] The present invention will be further described below in conjunction with specific examples. It should be understood that the following examples are only used to illustrate the present invention, rather than to limit the scope of the present invention.

[0038] Vitamin C is an existing commercial raw material purchased from Shanghai Bio-Tech Biotechnology Co., Ltd., product number ST1434.

[0039] Vitamin E is an existing commercial raw material purchased from Shanghai Bio-Tech Biotechnology Co., Ltd., product number S0079.

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

[0041] Fructus Ligustri Lucidi polysaccharide is an existing 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 embodiments 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 2g of Angelica polysaccharide and 2g of Ligustrum lucidum polysaccharide to 200mL 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, 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 above 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 easy to dissolve.

[0047] (3) 2g of vitamin E is first mixed with 1g of a food-grade emulsifier (such as lecithin) and 1mL of vegetable oil, and a uniform emulsion is formed by high shear stirring or ultrasonic dispersion. Then it 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 vitamin E in a relatively uniform and stable dispersed 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 water 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 made 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 effect 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) Compound vitamin group: a complex of vitamin C and vitamin E, with a weight ratio of 5:1; (3) Chinese medicine 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 them.

[0053] Escherichia coli (Beina Biotech, catalog number BNCC336902) and Shigella flexneri (Beina Biotech, catalog number BNCC232380) were selected and cultured in nutrient broth (LB) liquid medium (Shanghai Biotech Co., Ltd., catalog number ST163) at 37°C overnight. Then, the bacterial concentration was adjusted to 1.0 McFarland concentration with physiological saline to obtain bacterial suspension.

[0054] 3. Bacterial stimulation of peripheral blood experiment

[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 6 healthy adult subjects using glass tubes containing lithium heparin anticoagulant. 500 μl of anticoagulated whole blood was transferred to a sterile glass tube without additives and divided into 4 groups, and the above-mentioned compositions of different groups were added respectively (blank control group, normal saline with the same volume as anticoagulated whole blood; multivitamin group, 60 μg / ml; Chinese medicine polysaccharide group, 20 μg / ml; combined medication group, 80 μg / ml), and mixed thoroughly.

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

[0058] Take 50 μl of the whole blood containing the above composition and bacterial suspension, spread it on a 10 cm plate containing nutrient broth (LB) solid culture medium (Shanghai Biyuntian Biotechnology Co., Ltd., product number ST163), culture it at 37°C overnight, count the number of bacterial monoclonal colonies on the plate, and thus 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 Colony growth of Shigella flexneri is shown. Figure 4 The relative number of viable bacteria from a Shigella flexneri culture experiment is shown. Figures 1 to 4 It shows that the multivitamin group significantly promotes peripheral blood sterilization, the Chinese medicine polysaccharide group has no obvious effect, and the combination group significantly promotes peripheral blood sterilization. Compared with the multivitamin group, the combination group has a more significant effect on promoting peripheral blood sterilization. This shows that the antibacterial composition of the present invention has a good bactericidal effect, especially on Escherichia coli and Shigella flexneri. In addition, the antibacterial composition of the present invention can also have a significant inhibitory effect on 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 medicine polysaccharide group.

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

[0061] The results of IL-6 cytokine concentration test are as follows Figure 5 and Figure 6 As shown. Among them, Figure 5 Shown is the IL-6 cytokine concentration in plasma after the addition of E. coli. Figure 6The concentration of IL-6 cytokine in plasma after adding Shigella flexneri is shown. The results show that the multivitamin group significantly inhibited the secretion of IL-6, an inflammatory factor in peripheral blood induced by bacteria, the Chinese medicine polysaccharide group had no significant effect, and the combined drug group significantly inhibited the secretion of IL-6, an inflammatory factor in peripheral blood induced by bacteria. Compared with the multivitamin group, the combined drug group inhibited the secretion of IL-6, an inflammatory factor in peripheral blood induced by bacteria, more significantly.

[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 invention 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.

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) adding angelica polysaccharide and Ligustrum lucidum polysaccharide into water and stirring to dissolve; (2) adding vitamin C powder and stirring until completely dissolved to obtain a mixed solution; (3) Mixing vitamin E emulsifier and vegetable oil, stirring evenly, and then slowly adding the vitamin E to the mixed solution, stirring and homogenizing to make it evenly dispersed 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 and / or fungi.

9. The use according to claim 8, characterized in that: 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.

10. Use of the antibacterial composition according to any one of claims 1 to 6 in the preparation of a drug for treating inflammation.

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