Perilla frutescens extract having antibacterial efficacy of porphyromonas gingivalis and use thereof

Abstract

The invention provides a perilla frutescens extract with an antibacterial effect of Porphyromonas gingivalis, wherein the perilla frutescens extract is obtained by extracting a whole plant of perilla frutescens, and the perilla frutescens extract has an antibacterial effect of Porphyromonas gingivalis. The invention also provides application of the perilla frutescens extract in preparation of the health-care composition for resisting porphyromonas gingivalis.

Description

Technical Field

[0001] The present invention provides a perilla plant extract having antibacterial effect on Porphyromonas gingivalis and uses thereof. Background Art

[0002] Perilla (Perilla frutescens) is a plant with rich nutritional and medicinal properties. Both the plant itself and its extracts are widely used in food and traditional medicine. Perilla is often cited for its antioxidant properties due to its rich content of flavonoids, polyphenols, and vitamin C, all of which help neutralize free radicals in the body and reduce cellular damage. Perilla leaves have a refreshing flavor and are ideal for eating raw. They are also often used in salads or as wraps for raw food.

[0003] Porphyromonas gingivalis is a Gram-negative bacterium that primarily lives in the gingival spaces, dental plaque, and other oral tissues. It is a common oral bacterium. It produces enzymes and toxins, such as gingipains and osteolytic proteases, which contribute to the destruction of teeth and gum tissue. Previous studies have confirmed that P. gingivalis is one of the main pathogens causing periodontal disease. It is also believed to interfere with the host's immune response, leading to chronic inflammation of the periodontal tissues. Other studies have shown that P. gingivalis genetic material can be detected in the brain tissue of patients with Alzheimer's disease, suggesting that it may enter the brain through the bloodstream or other mechanisms, causing symptoms such as dementia. Epidemiological statistics also show that patients with periodontal disease are more likely to suffer from Alzheimer's disease, indicating that Porphyromonas gingivalis has a certain degree of correlation between oral health and nervous system health.

[0004] There is an urgent need to develop an oral health care composition that inhibits oral pathogens, such as Porphyromonas gingivalis. Summary of the Invention

[0005] Therefore, the present invention provides a perilla extract having antibacterial activity against Porphyromonas gingivalis, which can be used to develop an oral health care composition for inhibiting Porphyromonas gingivalis.

[0006] In one aspect, the present invention provides a Perilla frutescens extract having antibacterial efficacy against Porphyromonas gingivalis, wherein the Perilla frutescens extract is extracted from the whole plant of Perilla frutescens.

[0007] According to an embodiment of the present invention, the extract is obtained by solvent-free microwave essential oil extraction.

[0008] According to an embodiment of the present invention, the composition of the extract comprises, by total weight, 0.5-1% of trans-2-pentenal, 3-5% of 2-ethylacrolein, 1-2% of cis-2-penten-1-ol, 1-5% of tert-butylcyclohexane, 1-5% of (E)-2-heptenal, 10-15% of cis-2,4-heptadienal, 25-35% of trans-2,4-heptadienal, 3-5% of limonene, 5-10% of l-perillaldehyde and 30-35% of vitamin E.

[0009] In another aspect, the present invention provides a use of a perilla extract for preparing a health-care composition for combating Porphyromonas gingivalis. For example, the health-care composition has the efficacy of preventing or inhibiting periodontal disease, promoting oral hygiene, or preventing Alzheimer's disease.

[0010] In another aspect, the present invention provides an oral health care composition having antibacterial efficacy against Porphyromonas gingivalis, comprising the perilla extract of the present invention and an oral health care acceptable carrier.

[0011] According to an embodiment of the present invention, the oral health care composition has the efficacy of preventing or inhibiting periodontal disease and cleaning the oral cavity.

[0012] According to an embodiment of the present invention, the oral care composition has the efficacy of preventing Alzheimer's disease.

[0013] According to an embodiment of the present invention, the oral care composition can be made into toothpaste, mouthwash or oral freshener.

[0014] It is to be understood that the foregoing summary and the following detailed description are exemplary and explanatory only and are not restrictive of the scope of the invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The foregoing summary of the invention and the following detailed description of the invention will be better understood when read in conjunction with the accompanying drawings. For the purpose of illustrating the present invention, there are shown in the accompanying drawings presently preferred embodiments.

[0016] Figure 1 Shown is the GC / MS chromatogram of the perilla extract of the present invention.

[0017] Figure 2 Shown is the distribution diagram of components contained in Perilla extract.

[0018] Figure 3 Shown are the antibacterial test results of the perilla extract of the present invention against Porphyromonas gingivalis.

[0019] The details of one or more embodiments of the present invention are set forth in the following description. Other features and advantages of the present invention will become apparent from the following detailed description of several specific embodiments and the appended claims. DETAILED DESCRIPTION

[0020] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which this invention belongs.

[0021] The present invention is further described by the following examples, which are provided for illustration and not for limitation.

[0022] As used herein, the article "a" or "an" refers to one or to more than one (ie, to at least one) of the grammatical object of the article. For example, "an element" refers to one element or more than one element.

[0023] The term "perilla" refers to a multifunctional plant that not only has a flavoring effect on food, but also has some medicinal value. Perilla includes two different species: green perilla (Perilla frutescens var. crispa) and red perilla (Perilla frutescens var. acuta). The perilla that can be used in the present invention includes the above two types. Perilla is a plant with green or purple-red leaves belonging to the Lamiaceae family. It is commonly used for food seasoning, especially in Asian cuisine, such as Japanese and Korean cuisine. In traditional Chinese medicine and traditional medicine, perilla is sometimes used to treat colds, indigestion, respiratory problems, etc.

[0024] The term "periodontal disease" refers to a disease of the mouth that typically involves infection and inflammation of the tissues surrounding the teeth. It is a chronic infection caused by bacteria that affects the tooth-supporting structures in the mouth, including the gums, alveolar bone, and the periodontal ligament surrounding the teeth. The primary cause of periodontal disease is the accumulation of dental plaque, but other factors may also affect the risk of developing the disease, including smoking, genetic factors, and diabetes. Periodontal disease is divided into two main stages: gingivitis and periodontitis.

[0025] The term "Alzheimer's disease" refers to a chronic, progressive neurological disease that typically develops in older adults. It is a type of dementia and one of the most common forms of Alzheimer's disease. It is caused by the gradual breakdown and death of nerve cells in the brain, leading to a gradual loss of cognitive function. Symptoms of Alzheimer's disease include cognitive decline, behavioral and personality changes, speech impairment, disorientation, decreased ability to function in daily life, and brain pathology.

[0026] According to the present invention, a Perilla frutescens extract having antibacterial efficacy against Porphyromonas gingivalis is provided.

[0027] The perilla extract of the present invention is obtained from the whole plant of Perilla frutescens and can be obtained using conventional standard extraction methods. In one embodiment, the extract is obtained by solvent-free microwave essential oil extraction. This method produces an extract with higher levels of bioactive substances than organic solvent extraction.

[0028] According to an embodiment of the present invention, the perilla extract comprises, by total weight, 0.5-1% of trans-2-pentenal, 3-5% of 2-ethylacrolein, 1-2% of cis-2-penten-1-ol, 1-5% of tert-butylcyclohexane, 1-5% of (E)-2-heptenal, 10-15% of cis-2,4-heptadienal, 25-35% of trans-2,4-heptadienal, 3-5% of limonene, 5-10% of l-perillaldehyde, and 30-35% of vitamin E.

[0029] According to an embodiment of the present invention, the perilla extract can be used to prepare a health care composition having antibacterial efficacy against Porphyromonas gingivalis.

[0030] Therefore, the present invention also provides an oral health care composition having antibacterial efficacy against Porphyromonas gingivalis, comprising the perilla extract of the present invention and an oral health care acceptable carrier.

[0031] According to the present invention, the perilla extract or its oral health care composition can be used to prevent or inhibit periodontal disease and oral hygiene, for example, to prepare toothpaste, mouthwash or oral freshener.

[0032] The term "oral health acceptable carrier" refers to an acceptable carrier, suspending agent, flow control agent, emulsifier, preservative, lubricant, diluent, or excipient, etc., in the sense that it is compatible with the other ingredients of the formulation and is not harmful to the individual to whom the oral health composition is administered. Oral health acceptable carriers include aqueous and non-aqueous liquid media and various solid and semi-solid dosage forms. In addition to the active agent, such carriers may also contain various ingredients and additives, such as stabilizing active agents, binders, etc.

[0033] The following non-limiting examples are provided to aid those skilled in the art in the art in implementing the present invention. These examples should not be construed as limiting the present invention. Those skilled in the art may modify and vary the embodiments discussed herein without departing from the spirit or scope of the present invention and remain within the scope of the present invention.

[0034] Example

[0035] Example 1 Preparation of Perilla Extract

[0036] A quantitative sample of Perilla frutescen plants, including leaves, was placed in a microwave natural product extractor (ETHOS X 2.0, Milestone, Italy) and subjected to solvent-free microwave extraction (SFME). This method, which relies on the absorption of microwaves by the water content of the extracted sample, disrupts the cells and extracts the essential oil.

[0037] Example 2 Analysis of components of perilla extract

[0038] (1) Test method

[0039] In this study, gas chromatography / mass spectrometry (GC / MS) was used to analyze the main components of the perilla extract, wherein the separation column HP-5MS (25 m×0.20 mm×0.33 mm) of the gas chromatograph was used.

[0040] The gas chromatograph temperature gradient was set as follows: initial analysis temperature 35.0°C, maintained for 3.0 min, increased by 5.0°C per minute, maintained at 200.0°C for 0 min, then increased by 15.0°C per minute to 280.0°C, maintained for 8.0 min, and the final column temperature was 270.0°C.

[0041] The injection settings for the perilla extract sample were as follows: injection volume of 1.0 μL, carrier gas of helium (99.9995%), injection port temperature of 270.0°C, ion source temperature of 230.0°C, splitless injection mode; carrier gas flow rate of 20.0 mL / min, column flow rate of 1.4 mL / min, and mass spectrometer temperature of 150.0°C.

[0042] After the experiment is completed, the mass spectra at different retention times are analyzed and compared with the mass spectrum database of the National Institute of Standards and Technology (NIST) for essential oil components.

[0043] (2) Results

[0044] The components of perilla extract were analyzed by GC / MS. Figure 1 , containing different compound peak segments, mainly composed of 10 components (as shown in Table 1), namely: (1) 0.64% trans-2-pentenal ((E)-2-Pentenal, 5.016min); (2) 3.07% 2-ethylacrolein (2-Ethylacrolein, 5.290min); (3) 1.47% cis-2-penten-1-ol ((Z)-2-Penten-1-ol, 5.714min); (4) 2.86% tert-butylcyclohexane (1,1-dimethylethyl-Cyclohexan, 6.654min); (5) 2.21% (E)-2-heptenal ((E)-2-heptenal ,11.910min);(6)13.46%cis-2,4-heptadienal((E,Z)-2,4-Heptadienal,13.254min);(7)30.38%trans-2,4-heptadienal((E,E)-2,4-Heptadienal,13.698min);(8)4.19%limonene(Limonene,14.306min);(9)8.62%l-perillaldehyde(1-Cyclohexene-1-carboxaldehyde,4-(1-methylethenyl),21.755min);(10)33.08%vitamin E(Vitamin E,42.439min).

[0045] Table 1. Analysis of components of perilla extract

[0046]

[0047] According to the results of principal component analysis in Table 1 above, the components of perilla extract are classified and distributed as follows: Figure 2 As shown, it includes: 49.17% aldehydes, 3.93% monoterpenes, 2.88% ketones, 2.07% alkenes, 5.77% alcohols, and 5.18% amines.

[0048] Table 2. Content of components of Perilla extract

[0049] Aldehydes Monoterpenes Ketones Olefins Alcohols Amines 49.17％ 3.93％ 2.88％ 2.07％ 5.77％ 5.18％

[0050] Example 3 Antibacterial Activity Test of Perilla Extract against Porphyromonas gingivalis

[0051] The perilla extract of the present invention was tested for antimicrobial activity against Porphyromonas gingivalis according to ASTM E2315-16, Standard Guide for Assessment of Antimicrobial Activity Using a Time-Kill Procedure.

[0052] The test was divided into (1) an experimental group comprising the perilla extract prepared according to Example 1, and (2) a control group comprising 0.85% sterile saline.

[0053] First, adjust the strain of Porphyromonas gingivalis ATCC 33277 to the desired concentration of approximately 1.0 x 108 to 1.0 x 109 CFU / mL. Add 0.1 mL of the suspension to 10 mL of the experimental and control groups. Incubate at 35°C ± 2°C for 10 days and then at 25°C ± 2°C for 4 minutes. Immediately after inoculation, perform 10-fold serial dilutions in 9 mL of SCLP broth. After serial dilutions, inoculate the suspension onto appropriate culture media, incubate the culture media, observe the growth, and record the colony count.

[0054] The antibacterial capacity was calculated using the following antibacterial capacity calculation formula:

[0055] Calculation formula for antibacterial ability

[0056] LR=Log(A)-Log(D)

[0057] Antibacterial rate (R) = 100% × (1-10-LR)

[0058] A: Average bacterial count of the control group immediately diluted.

[0059] D: Average bacterial count of the experimental group after dilution.

[0060] The test results are as follows Figure 3 As shown in Table 3, the perilla extract of the present invention has an antibacterial rate of 89.03%.

[0061] Table 3. Content of components of Perilla extract

[0062]

[0063] 1 Antibacterial rate (%): If the antibacterial rate is less than 0.00%, it will be expressed as 0.00%.

[0064] Table 4. Determination of test establishment conditions

[0065]

[0066] 1 Conditions for establishment: All conditions must be met (except when the action time is greater than 1 hour).

[0067] Those skilled in the art will appreciate that changes may be made to the above-described specific embodiments without departing from the broad inventive concept thereof. Therefore, it should be understood that the present invention is not limited to the particular embodiments disclosed, but is intended to cover modifications within the spirit and scope of the present invention as defined by the appended claims.

Claims

1. A Greenperilla frutescens extract having antibacterial efficacy against Porphyromonas gingivalis, wherein the Greenperilla frutescens extract is extracted from the whole plant of Greenperilla frutescens, and comprises, by total weight, 0.5-1% of trans-2-pentenal, 3-5% of 2-ethylacrolein, 1-2% of cis-2-penten-1-ol, 1-5% of tert-butylcyclohexane, 1-5% of (E)-2-heptenal, 10-15% of cis-2,4-heptadienal, 25-35% of trans-2,4-heptadienal, 3-5% of limonene, 5-10% of L-perillaldehyde, and 30-35% of vitamin E.

2. The perilla plant extract as claimed in claim 1, wherein the extract is obtained by solvent-free microwave essential oil extraction.

3. Use of a perilla extract for preparing a health-care composition for combating Porphyromonas gingivalis.

4. The use according to claim 3, wherein the health care composition has the efficacy of preventing or inhibiting periodontal disease and oral hygiene.

5. The use according to claim 3, wherein the health care composition has the efficacy of preventing Alzheimer's disease.

6. An oral health care composition having antibacterial efficacy against Porphyromonas gingivalis, comprising the perilla extract according to claim 1 or 2, and an oral health care acceptable carrier.

7. The oral care composition according to claim 6, wherein the extract has the efficacy of preventing or inhibiting periodontal disease and oral hygiene.

8. The oral care composition according to claim 6, wherein the extract has an effect of preventing Alzheimer's disease.

9. The oral care composition according to claim 6, wherein the oral care composition can be made into toothpaste, mouthwash or oral freshener.

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