CN111407773A - Natural biological anti-inflammatory antibacterial agent - Google Patents

Natural biological anti-inflammatory antibacterial agent Download PDF

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Publication number
CN111407773A
CN111407773A CN202010374364.1A CN202010374364A CN111407773A CN 111407773 A CN111407773 A CN 111407773A CN 202010374364 A CN202010374364 A CN 202010374364A CN 111407773 A CN111407773 A CN 111407773A
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inflammatory
natural biological
antibacterial agent
biological anti
extract
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CN202010374364.1A
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Chinese (zh)
Inventor
马旭岩
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Harbin Ruiyan Biomedical Technology Co ltd
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Harbin Ruiyan Biomedical Technology Co ltd
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Abstract

The natural biological anti-inflammatory antibacterial agent of the invention is composed of citric acid, zinc oxide, L-arginine, natural extracts and water, wherein the natural extracts are licorice extract, ginger extract, schisandra extract, coptis extract and green tea extract.

Description

Natural biological anti-inflammatory antibacterial agent
Technical Field
The present invention relates to an anti-inflammatory and antibacterial agent.
Background
Inflammation is a normal defense protective phenomenon in the body that occurs locally to tissue damage due to physical trauma, harmful chemicals, microbial infection, or irritants in metabolites in the body. This inflammation is caused by a variety of chemical mediators produced by damaged tissues and moving cells (migrating cells), which assume different states depending on the morphology of the inflammatory process, and in normal cases, the living body neutralizes or eliminates pathogenic factors by an inflammatory reaction and regenerates the damaged tissues to restore normal structure and function, but in other cases, develops into a disease state, such as chronic inflammation. In addition, if the stimulus is caused by an autoimmune response (e.g. asthma or rheumatoid arthritis) or by inappropriate triggering of harmless substances (e.g. pollen), the defense response itself may damage the tissue and thus require anti-inflammation. Inflammatory responses are observed in almost all clinical conditions, and some bacterial diseases among these inflammatory diseases can be etiologically treated by antibiotics, but most are tissue damage caused by autoimmune reactions, and are therefore referred to as problematic complications without special therapies.
The most common anti-inflammatory agents for the treatment of these inflammatory diseases are classified into steroidal and non-steroidal anti-inflammatory agents. Since most of these synthetic anti-inflammatory drugs are accompanied by many side effects in addition to their main functions, there is an urgent need for development of anti-inflammatory drugs with good efficacy and low side effects. That is, treatment of inflammatory conditions with appropriate non-steroidal anti-inflammatory drugs (NSAIDs) alleviates inflammation, using a steroid formulation if the inflammation is severe or without the efficacy of NSAIDs, and using an immunosuppressive or surgical therapy if the condition is a difficult complication. NSAIDs are mainly used for inhibiting Cyclooxygenase (COX) and thus inhibiting the production of prostaglandins (prostaglandins) involved in inflammatory reactions, thereby producing anti-inflammatory effects, but they also have side effects such as gastrointestinal disorders, hepatic disorders, and renal disorders, and thus are difficult to use for a long period of time. Compared to these NSAIDs, steroid formulations are the fastest available for use in patients and are called fast-acting, well-acting anti-inflammatory drugs. However, as is well known, this drug has very strong toxicity, which lowers its own resistance to bacterial infection, and also causes exacerbation of diabetes, renal failure, mental dysfunction, and the like. And are difficult to stop halfway once treatment is initiated, care must be taken in using such agents, if possible, to avoid them.
Since synthetic anti-inflammatory agents as such are often accompanied by various side effects, development of anti-inflammatory drugs which are "potent and less side-effects" as mentioned above has been desired in reality. From the viewpoint of therapeutic effects and side effects, natural preparations which have been clinically experienced and have been widely evaluated in terms of safety since ancient times will be very good candidates for the development of preventive and therapeutic agents for inflammatory diseases.
In addition, human skin is an organ that is in direct contact with the surrounding environment, and is susceptible to infection with bacteria, fungi, and the like, which are involved in various skin diseases. Antibacterial compounds of triclocarban and triclosan have been used as antibacterial substances against these bacteria, but they cause side effects on the skin such as burning and redness of the skin when used for a long time. Therefore, there is a need to develop a new drug having anti-inflammatory and antibacterial activities with less side effects.
Disclosure of Invention
The invention provides a natural biological anti-inflammatory antibacterial agent, aiming at solving the technical problem of great side effect of the existing anti-inflammatory antibacterial agent.
The natural biological anti-inflammatory antibacterial agent consists of citric acid, zinc oxide, L-arginine, a natural extract and water, wherein the natural extract is a liquorice extract, a ginger extract, a schisandra extract, a coptis extract and a green tea extract.
The natural biological anti-inflammatory antibacterial agent comprises, by mass, 0.01% -5% of citric acid, 0.0001% -5% of zinc oxide, 0.01% -5% of L-arginine, 0.01% -5% of a natural extract and the balance of water.
The anti-inflammatory and antibacterial composition comprises natural extracts, citric acid, zinc oxide, L-arginine and water, has the characteristics of obvious and efficient anti-inflammatory and antibacterial effects compared with the components contained independently, has the effects of blocking or inhibiting the generation of inflammatory factors, and can treat or relieve inflammation.
In the present invention, citric acid has not only excellent antioxidant and anti-inflammatory properties but also an effect of neutralizing acidified components in the human body.
Zinc oxide, citric acid and L-arginine together serve as active ingredients in the present invention to enhance anti-inflammatory and antibacterial activity.
L-arginine is an amino acid constituting a protein, which is not essential for adults but essential for infants, belongs to a protein present in fish sperm, in animals such as herring, salmon, etc., in addition to which about 70% of the constituting amino acids are arginine, L-arginine is present in a plant seed in a free form, and as a metabolic pathway in vivo, it is a component of the ornithine cycle found by H.A. Krebs et al, and is decomposed into urea and ornithine by arginase, and is produced from citrulline and aminosuccinic acid in vivo L-arginine plays a role in protecting the human body from the toxic action of ammonia or a large number of amino acids, and increases anti-inflammatory activity together with citric acid and zinc oxide.
The licorice extract, ginger extract, schisandra extract, coptis extract and green tea extract of the present invention are not limited to the extraction methods thereof. Can be prepared by hot liquid extraction, cold soaking extraction, heating extraction, circulation cooling extraction, ultrasonic extraction, etc. The solvent may be water, ethanol, or a mixed solvent thereof.
The natural biological anti-inflammatory antibacterial agent of the present invention can prevent and treat inflammation-related diseases by inhibiting the expression of inflammatory factors nitrite, interleukin-1 β, interleukin-6, TNF- α.
The natural biological anti-inflammatory antibacterial agent is used for inhibiting the generation of inflammatory factors and also used for inhibiting escherichia coli, staphylococcus aureus and candida.
The natural biological anti-inflammatory antibacterial agent can be used as a medicinal component and can be prepared into oral medicines and external preparations according to an injection way.
When the pharmaceutical composition of the present invention is formulated into oral dosage forms, powders, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers and the like can be prepared with a suitable carrier according to methods known in the art. In this case, suitable pharmaceutically acceptable carriers include starch, lactose, glucose, sucrose, dextrose, sorbitol, mannitol, xylitol, corn starch, potato starch, wheat starch, methyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose and hydroxypropyl methyl cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyols, vegetable oils, and the like. The formulation may contain diluents such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like, or excipients, as required by the formulation.
When the pharmaceutical composition of the present invention is prepared into external preparations, it can be formulated in the form of injections, transdermal injections, nasal inhalants and suppositories, together with suitable carriers according to methods known in the art. When formulated as an injection, suitable carriers include sterile water, ethanol, polyhydric alcohol (e.g., glycerol or propylene glycol) or mixtures thereof, preferably ringer's solution, triethanolamine-containing PBS (phosphate buffered saline) or sterile water for injection, isotonic solution like 5% Textrose, etc. When formulated into transdermal injection, it can be formulated into ointment, cream, lotion, gel, topical solution, Pasta agent, liniment, aerosol, etc. In the case of nasal inhalants, dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide may be formulated into an aerosol form using a suitable propellant. When formulated into suppositories, witepsol, tween61, polyethylene glycols, cacao butter, taurine resins, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters and the like can be used as the base.
The injection amount of the pharmaceutical composition of the present invention is determined according to the condition, body weight, sex, age, severity of the patient, and injection route of the patient, and may be 0.001mg/kg to 10g/kg, preferably 0.001mg/kg to 1g/kg, for 1 day. Injections can be divided into 1 or several injections per day.
In addition, the natural biological anti-inflammatory antibacterial agent is applied to preparing food, has anti-inflammatory and antibacterial functions, contains stable physiologically active substances or minerals, and is used as food in the form of health functional food, nutritional supplement, health food or food additive. The natural biological anti-inflammatory antibacterial agent of the present invention can be produced in any form, for example, drinks such as tea, fruit juice, carbonated beverage and ionic beverage; processed milk such as milk and yogurt; foods such as chewing gum, rice cake, Korea fruit, bread, biscuit, and flour; can be made into health functional food preparations such as capsule, pill, granule liquid, powder, tablet, paste, syrup, gel, jelly, etc.
In addition, the natural biological anti-inflammatory antibacterial agent is applied to cosmetics and can relieve inflammatory skin irritation. The natural biological anti-inflammatory antibacterial agent of the present invention is used as a cosmetic composition, and further comprises ingredients commonly used in cosmetic compositions, such as stabilizers, solubilizers, surfactants, vitamins, pigments, and conventional adjuvants. The cosmetic composition of the present invention may be manufactured in any formulation conventionally prepared in the art, such as, but not limited to, a solution, a suspension, an emulsion, a cream, a gel, a cream, an emulsion, a powder, a soap, a surfactant-containing cleanser, an essential oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray. More specifically, it can be made into smoothing toner, nutrient solution, nourishing cream, massage cream, essence, eye cream, makeup removing cream, facial cleanser, makeup removing lotion, facial mask, spray or powder.
The natural biological anti-inflammatory antibacterial agent is prepared into an external agent in a pharmaceutical composition or a cosmetic composition, and further comprises 0.0001-0.1 mass percent of propolis wax, 0.1-1 mass percent of glycerol and 0.0001-0.1 mass percent of octanediol. The above effective components can further enhance antiinflammatory and antibacterial effects, and also have moisturizing and skin irritation relieving effects.
Drawings
FIG. 1 is a graph showing the cytotoxicity results of test two;
FIG. 2 is a graph showing the results of an assay for the inhibitory activity of trinitrate production;
FIG. 3 is a graph showing the results of an assay for the inhibitory activity of TNF- α in assay four;
FIG. 4 is a graph showing the results of the test for the development inhibitory activity of I L-1 β in test IV;
FIG. 5 is a graph showing the results of the inhibitory activity assay for I L-6 in assay four;
fig. 6 is a photograph of the test object in test five.
Detailed Description
The first embodiment is a natural biological anti-inflammatory antibacterial agent, which is composed of citric acid, zinc oxide, L-arginine, natural extracts and water.
The second embodiment is different from the first embodiment in that the natural biological anti-inflammatory and antibacterial agent comprises, by mass, 0.01-5% of citric acid, 0.0001-5% of zinc oxide, 0.01-5% of L-arginine, 0.01-5% of a natural extract, and the balance of water.
The third concrete implementation mode: the present embodiment differs from the first or second embodiment in that: the natural extracts are licorice extract, ginger extract, schisandra extract, coptis extract and green tea extract. The others are the same as in the first or second embodiment.
The fourth concrete implementation mode: the third difference between the present embodiment and the specific embodiment is that: the mass fractions of the five natural extracts are equal. The rest is the same as the third embodiment.
The fifth concrete implementation mode: the embodiment is an application of the natural biological anti-inflammatory antibacterial agent in the first embodiment, and the natural biological anti-inflammatory antibacterial agent is used as a medicine component.
The sixth specific implementation mode: the fifth embodiment is different from the fifth embodiment in that: the medicine can be prepared into oral medicines and external preparations according to the injection route. The rest is the same as the fifth embodiment.
The seventh embodiment: the fifth embodiment is different from the fifth embodiment in that: the dosage of the natural biological anti-inflammatory antibacterial agent is 0.001 mg/kg-10 g/kg in 1 day. The rest is the same as the fifth embodiment.
The specific implementation mode is eight: the embodiment is an application of the natural biological anti-inflammatory antibacterial agent in the first embodiment, and the natural biological anti-inflammatory antibacterial agent is used as a food ingredient.
The specific implementation method nine: the embodiment is an application of the natural biological anti-inflammatory antibacterial agent in the first embodiment, and the natural biological anti-inflammatory antibacterial agent is used as a cosmetic component.
The detailed implementation mode is ten: this embodiment differs from one of the sixth or ninth embodiments in that: when the natural biological anti-inflammatory antibacterial agent is used as a medicine external agent or a cosmetic ingredient, the natural biological anti-inflammatory antibacterial agent also comprises 0.0001-0.1 mass percent of propolis wax, 0.1-1 mass percent of glycerin and 0.0001-0.1 mass percent of octanediol. The above effective components can further enhance antiinflammatory and antibacterial effects, and also have moisturizing and skin irritation relieving effects. The others are the same as in one of the sixth or ninth embodiments.
The invention was verified with the following tests:
test I, the test is a natural biological anti-inflammatory antibacterial agent, which comprises the following components of 0.3 percent of citric acid, 0.007 percent of zinc oxide, 0.07 percent of L-arginine, 1 percent of natural extract and the balance of water according to mass fraction;
the natural extracts are licorice extract, ginger extract, schisandra extract, coptis extract and green tea extract, and the mass of the five components is equal.
The preparation method of the natural biological anti-inflammatory antibacterial agent comprises the following steps: all the raw materials are weighed and mixed evenly, stirred for 1h at 50 ℃, cooled to room temperature, then dried and powdered.
Test two, this test is a cell culture test in which RAW264.7 cells are cultured at 1.0 × 106CellIs/m L were dispensed into 24 microliter plates at 37 ℃ with 5% CO2After incubation for 18h, test one prepared natural biological anti-inflammatory antibacterial agent and L PS (1. mu.g/m L) were added simultaneously and incubated for 24h, after incubation, MTT was added at a concentration of 500ug/m L, reaction was carried out at 37 ℃ for 3h, then the supernatant was removed, the resulting Formazan sediment was dissolved after DMSO was added to react with the live cells, and then transferred to a 96-well plate and absorbance was measured at 540 nm.
The cytotoxicity results of test two are shown in fig. 1, and the abscissa is the amount of the natural anti-inflammatory agent prepared by adding test one thereto, and it can be seen from fig. 1 that the natural anti-inflammatory agent prepared by adding test one does not show any specific cytotoxicity.
And (3) test III: this assay is an anti-inflammatory activity assay:
NO production inhibitory activity test:
RAW264.7 cells were cultured at 1.0 × 106CellIs/m L were dispensed into 24 microliter plates at 37 ℃ with 5% CO2After incubation for 18h under conditions of (1 μ g/m L), test one prepared natural biological anti-inflammatory antibacterial agent and L PS (1 μ g/m L) were added simultaneously, and incubated for 24h, after which cell culture supernatant 100u L and Griess reagent 100u L were mixed together, reacted in a 96-well plate for ten minutes, and absorbance was measured at 540 nm. the nitrite content in the sample was quantified using standard curves of sodium nitrite dissolved in the same medium at various concentrations, as shown in FIG. 2, the abscissa is the amount of natural biological anti-inflammatory antibacterial agent prepared by addition of test one, and as shown in the drawing, it was confirmed that the treated sample inhibited the production of nitrite according to concentration.
The test is the expression inhibition activity test of interleukin-1 β, interleukin-6 and tumor necrosis factor-a:
RAW264.7 cells were cultured at 1.0 × 106CelIs/m L to 24 μIn a liter plate, 5% CO at 37 ℃2After incubation for 18h, test one prepared natural biological anti-inflammatory antibacterial agent and L PS (1 μ g/m L) were added simultaneously and incubated for 24h, and after the incubation was completed, the culture medium was centrifuged to obtain supernatant I L-1 β, 1L-6 and TNF- α, which were quantified using E L ISA kit.
The results of the TNF- α inhibition activity assay are shown in FIG. 3.
The results of the test for the development inhibitory activity of I L-1 β are shown in FIG. 4.
The results of the inhibition activity test of I L-6 are shown in FIG. 5.
As shown in the figure, the treated samples were confirmed to inhibit the occurrence of L-1 β, 1L-6 and TNF- α in concentration dependence, and in addition, had similar level of inhibitory effect as the anti-inflammatory component DEX.
And (5) testing: this test is an antibacterial activity test: the results of the antibacterial activities against escherichia coli, staphylococcus aureus and candida were shown in table 1 and fig. 6 below, in which the three graphs in the first row of fig. 6 are blank group photos of escherichia coli, staphylococcus aureus and candida from left to right in sequence, and the three graphs in the second row are test group photos of escherichia coli, staphylococcus aureus and candida from left to right in sequence.
TABLE 1
Figure BDA0002479545820000071
As shown in tables 1 and 6, it was confirmed that the natural biological anti-inflammatory agent of test one has excellent antibacterial activity against Escherichia coli, Staphylococcus aureus and Candida.

Claims (10)

1. A natural biological anti-inflammatory antibacterial agent is characterized in that the natural biological anti-inflammatory antibacterial agent is composed of citric acid, zinc oxide, L-arginine, natural extracts and water.
2. The natural biological anti-inflammatory and antibacterial agent as claimed in claim 1, wherein the natural biological anti-inflammatory and antibacterial agent comprises, by mass, 0.01-5% of citric acid, 0.0001-5% of zinc oxide, 0.01-5% of L-arginine, 0.01-5% of natural extract, and the balance of water.
3. The natural biological anti-inflammatory and antibacterial agent as claimed in claim 1 or 2, wherein the natural extract is licorice extract, ginger extract, schisandra extract, coptis extract and green tea extract.
4. The natural biological anti-inflammatory and antibacterial agent as claimed in claim 3, wherein the natural extracts comprise equal mass fractions of licorice extract, ginger extract, schisandra extract, coptis extract and green tea extract.
5. Use of a natural biological anti-inflammatory agent according to claim 1, characterised in that the natural biological anti-inflammatory agent is used as a pharmaceutical ingredient.
6. The use of a natural biological anti-inflammatory and antibacterial agent as claimed in claim 5, wherein said medicament is formulated into oral and topical preparations; when the medicine is used as a medicine external preparation, the medicine also comprises 0.0001-0.1 mass percent of propolis wax, 0.1-1 mass percent of glycerin and 0.0001-0.1 mass percent of octanediol.
7. The use of a natural antiinflammatory biocide according to claim 6, wherein the natural antiinflammatory biocide is used in an amount of: 0.001 mg/kg-10 g/kg in 1 day.
8. The use of a natural biological anti-inflammatory agent as claimed in claim 1, wherein said natural biological anti-inflammatory agent is used as a food ingredient.
9. Use of a natural biological anti-inflammatory antibacterial agent according to claim 1, characterized in that said natural biological anti-inflammatory antibacterial agent is used as a cosmetic ingredient.
10. The use of a natural biological anti-inflammatory antibacterial agent according to claim 9, characterized in that the natural biological anti-inflammatory antibacterial agent further comprises 0.0001-0.1% by mass of propolis wax, 0.1-1% by mass of glycerin, and 0.0001-0.1% by mass of octanediol when used as a cosmetic ingredient.
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