KR102598905B1 - Deodorant composition comprising Quillaia saponaria - Google Patents

Abstract

This relates to a deodorizing composition containing quillaja extract. According to one aspect, a composition containing quillaja extract as an active ingredient is not only used as a deodorant through its excellent deodorizing effect, but also can be recycled because it uses quillaja extract, a natural plant. Its effectiveness and safety have been recognized, making it useful in everyday life and various industrial fields.

Description

Deodorant composition comprising Quillaia saponaria}

It relates to a deodorizing composition containing Quillaja extract.

Quillaia saponaria M. , also called soap tree, is an evergreen tree of the Rosaceae family distributed in Chile, Bolivia, and Peru in South America. South American natives peeled the bark of a tree, soaked it in water, and used it like soap for washing or washing hair. It has been used as a foaming agent in beverages, especially non-alcoholic beverages and shake drinks, in many countries. In addition to food uses, Quillaja extract is widely used in various fields such as food, cosmetics, pharmaceuticals, detergents, building materials, and wastewater treatment. Most of the bioactive effects of Quillaja known so far are due to its active ingredients, triterpenoid saponins, of which Quillaja sapogenin and Quillaic acid are the main ingredients. These saponin components are known to have various beneficial functions such as surface activity, hemolysis, antibacterial activity, animal growth promotion, and complex formation with cholesterol. Due to these functions, it is used as a functional additive in various industrial fields such as feed, medicine, environment, and cosmetics (Waller, G., Yamasaki, K., Saponins used in traditional and Modern medicine, 1996).

Coronavirus (CoV) is a virus that can infect humans and various animals and is an RNA virus with a gene size of 27 to 32 kb. The Coronaviridae family can be classified into four genera (alpha, beta, gamma, and delta). Alpha and Beta can infect humans and animals, while Gamma and Delta can only infect animals. The shape, like the name of the coronavirus, appears spherical when observed under an electron microscope, and the external spike protein is characterized in the form of a crown. There are currently six types of human-infecting coronaviruses known. Types that cause colds include 229E, OC43, NL63, and HKU1, and types that can cause severe pneumonia include SARS-CoV and MERS-CoV. And the new coronavirus that has emerged since 2019 is SARS-CoV-2, which causes various respiratory infections ranging from mild to severe, such as fever, malaise, cough, difficulty breathing, and pneumonia. Other symptoms include phlegm, sore throat, headache, hemoptysis, nausea, and diarrhea. It is classified as a new infectious disease syndrome, a class 1 infectious disease in Korea, and is classified as a statutory infectious disease. It is spread through droplets and contact. The incubation period is 1 to 14 days, and the global fatality rate is about 3.4% (WHO, 3.5 standard), and there is still no treatment (Korea Centers for Disease Control and Prevention, http://ncov.mohw.go.kr/).

Quillaja extract protects against viral infections such as Reovirus (gastroenteritis), Vaccinia Virus (smallpox), VZV (herpes zoster), HSV-1 (herpes), HIV-1 and HIV-2 (autoimmune disease), and Rotavirus (diarrhea-causing). (Michael R Roner et al, Antiviral activity ovtained from aqueous extracts of the Chilean soapbark tree, 2007, Michael R Roner et al, Prevention of rotavirus infections in vitro with aqueous extracts of Quillaja saponaria Molina). However, there has been no report on infection resistance to any type of coronavirus.

Waller, G.R. & Yamasaki, K. (1996) Saponins used in traditional and modern medicine, Plenum Press, New York, NY. Michael R. Roner et al., Journal of General Virology (2007), 88, 275-285 Michael R. Roner et al., Future Med Chem. (2010), 2(7), 1083-1097

One aspect is to provide an antiviral pharmaceutical composition containing Quillaia saponaria MOLIMA extract as an active ingredient.

Another aspect is to provide an antiviral external skin preparation containing Quillaia saponaria MOLIMA extract as an active ingredient.

Another aspect is to provide an antiviral health functional food composition containing Quillaia saponaria MOLIMA extract as an active ingredient.

Another aspect is to provide a quasi-drug composition for preventing virus attachment containing Quillaia saponaria MOLIMA extract as an active ingredient.

Another aspect is to provide an antiviral cosmetic composition containing Quillaia saponaria MOLIMA extract as an active ingredient.

Another aspect is to provide a deodorizing composition containing Quillaia saponaria MOLIMA extract as an active ingredient.

Another aspect is to provide a deodorizing cosmetic composition containing Quillaia saponaria MOLIMA extract as an active ingredient.

One aspect is to provide an antiviral pharmaceutical composition containing quillaja extract as an active ingredient.

In this specification, “Quillaja” may refer to an evergreen tree of the Quillajaceae family distributed in Chile, Bolivia, and Peru in South America. Its scientific name is Quillaia saponaria MOLINA , and it is native to Chile and mainly lives at 2000m above sea level. It grows to about 15-20m and has thick, dark bark and smooth, oval-shaped leaves that are 3-5cm long. It is also called the soap tree, and the inner bark contains saponin, a type of natural soap.

In this specification, “extract” refers to a result such as a liquid component obtained by immersing the target material in various solvents and then extracting it at room temperature or at a heated state for a certain period of time, or a solid content obtained by removing the solvent from the liquid component. You can. In addition, in addition to the above results, it can be comprehensively interpreted to include all dilutions of the above results, concentrates thereof, crude preparations and purifications thereof, etc.

The extract may be Quillaja extract. Extraction solvents include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, liquid diluent, ethoxylated isostearyl alcohol. , suspending agents such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, Isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linoline derivative, or ethoxy. It may include sylated glycerol fatty acid ester, etc. Specifically, the extraction solvent may be a polar solvent, a non-polar solvent, or a combination thereof. The polar solvent includes water, anhydrous or hydrous alcohol having 1 to 6 carbon atoms (methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol or 2-butoxyethanol, etc.), or It may be an alcoholic compound (ethylene glycol, propylene glycol, or butylene glycol, etc.), acetic acid, DMFO (dimethyl-formamide), DMSO (dimethyl sulfoxide), or a combination thereof. Nonpolar solvents include hexane, oroalkane, pentane, hexane, Ethyl acetate, methyl acetate, butyl acetate, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, acetone, 1-chloropentane, o-xyl Ren, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, diethyl ether, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline , diethylamine, ether, carbon tetrachloride, THF, or a combination thereof. In this specification, the extract is not particularly limited thereto, and may include extracts showing substantially the same effect prepared using other solvents as well as the above solvent.

In one embodiment, the extraction method using the solvent may be performed according to a method known in the art. For example, a cold needle extraction method in which quillaya is immersed in the above solvent and extracted at a room temperature of 10 to 25°C, a heat extraction method in which the extract is extracted by heating to 40 to 100°C, a reflux extraction method using a reflux condenser, an ultrasonic extraction method using an ultrasonic device, and It can be performed through methods such as high-frequency extraction using a high-frequency device.

In one embodiment, the virus may be a coronavirus. The coronaviruses include alphacoronavirus, betacoronavirus, gammacoronavirus, deltacoronavirus, human coronavirus 229E (HCoV-229E), human coronavirus OC43 (HCoV-OC43), and severe acute respiratory syndrome coronavirus (SARS-CoV). ), human coronavirus NL63 (HCoV-NL63, New Haven coronavirus), human coronavirus HKU1, Middle East respiratory syndrome coronavirus (MERS-CoV) or severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2 or 2019 novel coronavirus) or 2019-nCoV).

As used herein, “antiviral” may mean the ability to inhibit the production of coronavirus particles, that is, the ability to effectively inhibit the formation and/or replication of coronaviruses in host cells, especially mammals including humans. Compositions containing the quillaja extract as an active ingredient can have an antiviral effect by interfering with the mechanisms necessary for viral infection, replication, or proliferation.

According to one embodiment, the content of the quillaja extract in the above or the following composition is 0.0001 to 100% by weight, 0.001 to 100% by weight, 0.01 to 100% by weight, 0.1 to 100% by weight, 1 to 100% by weight, 10 to 100% by weight. %, 0.0001 to 80% by weight, 0.001 to 80% by weight, 0.01 to 80% by weight, 0.1 to 80% by weight, 1 to 80% by weight, 10 to 80% by weight, 0.0001 to 70% by weight, 0.001 to 70% by weight, 0.01 to 70% by weight, 0.1 to 70% by weight, 1 to 70% by weight, 10 to 70% by weight, 0.0001 to 60% by weight, 0.001 to 60% by weight, 0.01 to 60% by weight, 0.1 to 60% by weight, 1 to 60% by weight, 10 to 60% by weight, 0.0001 to 50% by weight, 0.001 to 50% by weight, 0.01 to 50% by weight, 0.1 to 50% by weight, 1 to 50% by weight, 10 to 50% by weight, 0.0001 to 40% by weight %, 0.001 to 40% by weight, 0.01 to 40% by weight, 0.1 to 40% by weight, 1 to 40% by weight, 10 to 40% by weight, 0.0001 to 30% by weight, 0.001 to 30% by weight, 0.01 to 30% by weight, 0.1 to 30% by weight, 1 to 30% by weight, 10 to 30% by weight, 0.0001 to 20% by weight, 0.001 to 20% by weight, 0.01 to 20% by weight, 0.1 to 20% by weight, 1 to 20% by weight, 10 to It may be 20% by weight, 0.0001 to 10% by weight, 0.001 to 10% by weight, 0.1 to 10% by weight, or 1 to 10% by weight.

The number of times the composition according to one embodiment is administered is 1 to 3 times a day, or may be administered more times as needed. However, the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, gender, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity, and may be prescribed by those skilled in the art. If so, the dosage can be adjusted appropriately by taking these factors into consideration. The number of administrations can be one time or two or more times within the range of clinically acceptable side effects, and the administration site can be administered at one or two or more places. For animals other than humans, the dosage per kg is the same as that for humans, or the above dosage is converted into, for example, the volume ratio (e.g., average value) of the organs (heart, etc.) between the target animal and human. One dose can be administered. Possible routes of administration include oral, sublingual, parenteral (e.g., subcutaneous, intramuscular, intraarterial, intraperitoneal, intrathecal, or intravenous), rectal, topical (including transdermal), inhalation, and injection, or implantable. It may involve insertion of a device or substance. Examples of animals subject to treatment according to one embodiment include humans and other mammals, specifically humans, monkeys, mice, rats, rabbits, sheep, cows, dogs, horses, pigs, etc. Included. However, the dosage, frequency, and subject are not intended to limit the scope of the present specification.

The pharmaceutical composition according to one embodiment may include a pharmaceutically acceptable carrier and/or additive. For example, it contains sterile water, physiological saline, common buffers (phosphoric acid, citric acid, other organic acids, etc.), stabilizers, salts, antioxidants (ascorbic acid, etc.), surfactants, suspending agents, isotonic agents, or preservatives. can do. For topical administration, it may also include combinations with organic materials such as biopolymers and inorganic materials such as hydroxyapatite, specifically collagen matrices, polylactic acid polymers or copolymers, polyethylene glycol polymers or copolymers, and chemical derivatives thereof. You can.

According to one embodiment, the pharmaceutical composition may be used as a suspending agent, solubilizing agent, stabilizer, isotonic agent, preservative, anti-adsorption agent, surfactant, diluent, excipient, pH adjuster, analgesic agent, if necessary depending on the administration method or dosage form. , buffering agents, reducing agents, antioxidants, etc. may be appropriately included. The pharmaceutical composition according to one embodiment is formulated in unit dosage form using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by a person skilled in the art to which the invention pertains. It can be manufactured by or by placing it in a multi-capacity container. The formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of powder, granules, tablets or capsules.

Another aspect is to provide an antiviral external skin preparation containing Quillaja extract as an active ingredient.

At this time, the meanings of quillaja, extract, virus and antiviral are as described above.

In this specification, “external preparation” may be a preparation provided for external use, and an external skin preparation composition may be formulated containing a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, such as solutions, gels, solids, pasty anhydrous products, emulsions obtained by dispersing the oil phase in the water phase, suspensions, microemulsions, microcapsules, microgranules or ionic forms (liposomes) and It may be provided in the form of a non-ionic vesicular dispersion, or in the form of a cream, skin, lotion, powder, ointment, spray, pack or concealer stick. It may be used in the form of foam or in the form of an aerosol composition further containing compressed propellant, but is not limited thereto. These compositions can be prepared according to conventional methods in the art.

The composition for external skin application may have an antiviral effect.

Another aspect is to provide an antiviral health functional food composition containing Quillaja extract as an active ingredient.

At this time, the meanings of quillaja, extract, virus and antiviral are as described above.

In this specification, “food” refers to meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, It can be a vitamin complex, health functional food, health food, etc., and can include all foods in the conventional sense.

The food composition according to one embodiment can be consumed on a daily basis and can be expected to have excellent antiviral effects, so it can be very useful for health promotion purposes.

In this specification, "functional food" is the same term as food for special health use (FoSHU), and is a medical product processed to efficiently exhibit bioregulatory functions in addition to providing nutrition. It may mean food with high medical effects. Here, ‘function’ may mean controlling nutrients for the structure and function of the human body or obtaining useful effects for health purposes such as physiological effects. In this specification, food can be manufactured by methods commonly used in the industry, and can be manufactured by adding raw materials and ingredients commonly added in the industry. Additionally, the food formulation can be manufactured without limitation as long as it is a formulation recognized as a food. In this specification, the composition for food can be manufactured in various types of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long time because it is made from food as a raw material, and is excellent in portability, so it can be used for daily use. The food according to the embodiment can be consumed as an adjuvant to enhance the antiviral effect.

The above-mentioned health food may refer to food that has a more active health maintenance or promotion effect compared to general food, and health supplement food may refer to food for the purpose of health supplementation. In some cases, the terms health functional food, health food, or health supplement may be used interchangeably.

The health functional food is a food manufactured by adding the composition to food materials such as beverages, teas, spices, gum, or confectionery, or by encapsulating, powdering, or suspending it, and has a specific health effect when consumed. However, unlike regular drugs, it may have the advantage of not having any side effects that may occur when taking the drug for a long time since it is made from food.

The health functional food composition may further include a physiologically acceptable carrier. The type of carrier is not particularly limited, and any carrier commonly used in the art can be used.

In addition, the health functional food composition may include additional ingredients that are commonly used in food compositions to improve smell, taste, vision, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, and panthotenic acid. Additionally, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr); and amino acids such as lysine, tryptophan, cysteine, and valine.

In addition, the food composition contains preservatives (potassium sorbate, sodium benzoate, salicylic acid and sodium dehydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder and sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA) and butylhydroxide) roxitoluene (BHT), etc.), coloring agents (tar color, etc.), coloring agents (sodium nitrite and sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG and monosodium glutamate, etc.), sweeteners (dulcine, cyclemate, etc.) saccharin and sodium, etc.), flavorings (vanillin and lactones, etc.), leavening agents (alum and D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (grease), coating agents, gum base agents, anti-foam agents, solvents or improvers, etc. May contain food additives. The additives can be selected depending on the type of food and used in an appropriate amount.

The food composition according to one embodiment may be a health drink composition. The health drink composition may contain various flavoring agents or natural carbohydrates as additional ingredients like ordinary drinks. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; polysaccharides such as dextrins and cyclodextrins; It may be a sugar alcohol such as xylitol, sorbitol, or erythritol. Sweeteners include natural sweeteners such as thaumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be 0.01 to 0.04 g, specifically about 0.02 to 0.03 g, per 100 ml of the health drink composition of the present invention, but is not limited thereto.

In addition to the above, the health drink composition includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohol or carbonating agent. In addition, it may contain pulp for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These ingredients can be used independently or in combination. The ratio of these additives is not very important, but may be selected in the range of 0.01 to 1.0 parts by weight per 100 parts by weight of the health drink composition of the present invention.

Another aspect is to provide a quasi-drug composition that prevents virus attachment containing Quillaja extract as an active ingredient.

At this time, the meaning of Quillaja, extract and virus is as described above.

In this specification, “quasi-drugs” may refer to products that have a milder effect than pharmaceuticals among products used for the purpose of diagnosing, treating, improving, alleviating, treating, or preventing diseases in humans or animals. For example, according to the Pharmaceutical Affairs Act of the Republic of Korea, quasi-drugs exclude products used for medicinal purposes, and may include products used to treat or prevent diseases in humans and animals, and products that have a mild or no direct effect on the human body.

The quasi-drug composition may be manufactured in a form selected from the group consisting of body cleanser, foam, soap, mask, ointment, cream, lotion, essence, and spray, but is not limited thereto.

The quasi-drug composition can be used by adding the composition containing the quillaja extract as an active ingredient as is or with other quasi-drugs or quasi-drug components, and can be used appropriately according to conventional methods.

Another aspect is to provide an antiviral cosmetic composition containing Quillaja extract as an active ingredient.

At this time, the definitions of quillaja, extract, virus and antiviral are as described above.

The cosmetic composition includes solutions, external ointments, creams, foams, nutritional lotions, softening lotions, skin softeners, skin toners, astringents, perfumes, packs, softeners, emulsions, makeup bases, essences, soaps, liquid cleansers, bath additives, and sunscreens. Cream, sun oil, suspension, emulsion, paste, gel, lotion, milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, foundation, essence, nutritional essence, pack, powder, soap, Formulations selected from the group consisting of surfactant-containing cleansers, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, oils, powder foundations, emulsion foundations, wax foundations, patches, mask packs, aerosols, sprays and capsules. It can be manufactured, but is not limited to this.

The cosmetic composition may further include one or more cosmetically acceptable carriers that are blended with general skin cosmetics, and common ingredients include, for example, oil, water, surfactants, moisturizers, lower alcohols, thickeners, and chelating agents. , colorants, preservatives, fragrances, etc. may be appropriately mixed, but are not limited thereto. Cosmetically acceptable carriers included in the cosmetic composition may vary depending on the formulation.

When the formulation of the cosmetic composition is ointment, paste, cream or gel, the carrier ingredients include animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide. Or a mixture thereof can be used.

When the formulation of the cosmetic composition is powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silcate, polyamide powder, or mixtures thereof may be used as carrier ingredients, and especially in the case of spray, additional May contain propellants such as chlorofluorohydrocarbons, propane/butane or dimethyl ether.

When the formulation of the cosmetic composition is a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent may be used as a carrier component. For example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, and 1,3-butyl glycol oil can be used, especially cotton seed oil, peanut oil, corn germ oil, and olive oil. , castor oil and sesame oil, glycerol fatty esters, polyethylene glycol or fatty acid esters of sorbitan can be used.

When the formulation of the cosmetic composition is a suspension, the carrier component includes water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.

When the formulation of the cosmetic composition is a capsule, it may be formulated in the form of an alginate capsule, agar capsule, gelatin capsule, wax capsule, or double capsule. Not limited.

In addition, the cosmetic composition contains commonly used auxiliaries, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic activators, preservatives, antioxidants, solvents, fragrances, fillers, blocking agents, pigments, odorants and dyes, in addition to the quillaja extract. may include.

Another aspect is to provide a deodorizing composition containing quillaja extract as an active ingredient.

At this time, the meaning of Quillaja and extract, the dosage of Quillaja extract in the composition, and the formulation of the composition are as described above.

Another aspect provides a deodorizing cosmetic composition containing Quillaja extract as an active ingredient.

At this time, the meaning of Quillaya, extract and cosmetics, the dosage of Quillaya extract in the composition, and the formulation of the composition are as described above.

According to one aspect, a composition containing Quillaya extract as an active ingredient is not only used as a deodorant through its excellent deodorizing effect, but also uses Quillaya extract, a natural plant, so its recyclability and safety are recognized, making it suitable for use in daily life and various industrial fields. It has an effect that can be useful.

Figure 1 is a graph showing the difference in total saponin content between Quillaja general extract and Quillaja high-frequency extract. (Example 1: Quillaja general extract; Example 2: Quillaja high-frequency extract)
Figure 2 is a graph showing the antiviral effect of Emodin.
Figure 3 is a graph showing the antiviral effect of the same amount of Quillaja high-frequency extract as Emodin.

Hereinafter, preferred embodiments are presented to facilitate understanding of the present specification. However, the following examples are provided only to make the present invention easier to understand, and the content of this specification is not limited by the following examples.

Example 1: Preparation of Quillaja general extract

Add 10 to 20 times the amount of purified water to the dried and shredded Quillaja raw materials, extract at 80 to 90 ℃ for 2 to 3 hours, and naturally cool to room temperature. The precipitate was removed using a centrifuge and the supernatant was collected, or the supernatant filtered through filter paper (Filter paper ADVANTEC, No. 2, 110 mm) was used as an extract.

Example 2: Preparation of Quillaja radiofrequency extract

After adding 10 to 20 times as much purified water to the dried and shredded Quillaja raw materials, using a high-frequency device (HS9760, Alltech Co., Ltd.), 0.2 to 2 MHz at a temperature of 25℃ to 35℃, 80 to 90 MHz for 1 to 10 minutes. It was manufactured by extraction under kJ/kg conditions. The precipitate was removed using a centrifuge, and the supernatant was used as an extract.

Experimental Example 1: Comparison of total saponin content of Quillaja general extract and Quillaja high-frequency extract

In order to compare and analyze the total saponin content of the Quillaja general extract of Example 1 and the Quillaya high-frequency extract of Example 2, an analysis was requested from Nature & Biotech Co., Ltd.

The test report issuance number for the Quillaja general extract according to Example 1 is NNB-2020-N0978, and the issuance date is September 11, 2020. In addition, the test report issuance number of the Quillaja high-frequency extract according to Example 2 is NNB-2020-N-0976, and the issuance date is September 11, 2020.

Figure 1 shows the total saponin content in the Quillaja extract according to the extraction method. While the total saponin content of the general extract was 2.67%, the total saponin content of the high-frequency extract was 3.59%, which increased the total saponin content in the extract by a whopping 36.46%. These results confirmed that the efficiency of the high-frequency extraction method was superior to that of the general extraction method.

Experimental Example 2: Skin safety effect of Quillaja high-frequency extract

To confirm the skin safety of the Quillaja high-frequency extract of Example 2, a primary skin irritation test was conducted using a skin patch.

Specifically, patches containing about 0.5 g each of purified water, Quillaja general extract, or Quillaja high-frequency extract were attached to the back area, removed after 24 hours, and skin reactions were visually determined after 30 minutes, 24 hours, and 48 hours. . The experiment was conducted on a total of 33 adult men and women (27 women, 6 men), and the judgment criteria were determined according to the standards of the International Contact Dermatitis Research Group (ICDRG) shown in Table 1 below.

Range of Mean Score Criteria 0.00 ≤ R ＜1.01 hypoallergenic 1.01 ≤ R ＜ 2.01 light stimulation 2.01 ≤ R ＜3.51 medium irritation 3.51 ≤ R strong stimulation

The experimental results are shown in Table 2 below.

test substance Mean Score Judgment Purified water 1.01 light stimulation Quillaja regular extract 0.55 hypoallergenic Quillaja High Frequency Extract 0.51 hypoallergenic

As shown in Table 2, it was confirmed that the Quillaja extract, as well as the general extract and the high-frequency extract, are hypoallergenic and safe for the skin.

Experimental Example 3: Deodorizing effect of Quillaja high-frequency extract and spray containing it

We sought to confirm the deodorizing effect of Quillaja high-frequency extract through gas detection tube method.

Specifically, it was confirmed by requesting the Korea Apparel Testing and Research Institute (KATRI). The test conditions provided by the requesting institution are as follows.

1) Sample input amount: 50 ml

2) Test gas: ammonia (NH ₃ )

3) Concentration of injected test gas: 100 ㎍/㎖

4) Test gas injection volume: 1.5 L

5)Test container: 3 L tetler bag

6) Test environment: temperature (20 ± 2) ℃, humidity (40 ± 5) % RH

7) Deodorization power (%) = [(Blank gas concentration - Sample gas concentration)/Blank gas concentration] x 100

The composition of the spray for the experiment is shown in Table 3 below.

No. ingredient Formulation Example 1 Comparative formulation example 1 One Quillaja High Frequency Extract 77.5 - 2 Butylene Glycol 15 15 3 1,2-Hexanediol 2.5 2.5 4 2,3-Butanediol 5 5 5 Water To 100 To 100

The experimental results are shown in Table 4 below.

Test time, minutes Deodorization power (%), gas detection tube method Quillaja High Frequency Extract Formulation Example 1 Comparative formulation example 1 30 > 99 > 99 89.7 60 > 99 > 99 91.5 90 > 99 > 99 94.4 120 > 99 > 99 97.2

As shown in Table 4, the Quillaja high-frequency extract and the composition containing it (Formulation Example 1) had the effect of removing more than 99% of odors for up to 120 minutes.

Experimental Example 4: Antiviral test of Quillaja high-frequency extract

To determine the antiviral activity of Quillaja high-frequency extract, screening was conducted for inhibitors of the SARS-CoV-2 spike that binds to ACE2.

Specifically, antiviral testing against Coronavirus 2 (SARS-CoV-2) was confirmed through Spike RBD (SARS-CoV-2): ACE2 Inhibitor Screening Assay Kit (Catalog # 79931). The Spike RBD (SARS-CoV-2) kit is composed as shown in Table 5 below.

Catalog # Component Amount 100684 SARS-CoV-2 Spike Protein (RBD), mFc Tag 5 ㎍ 11003 ACE2, His-Tag 5 ㎍ 79311 3x Immuno Buffer 1 50ml 79728 Blocking Buffer 2 50ml Anti-His-HRP 10㎕ 79670 ELISA ECL substrate A (Transparent bottle) 6ml ELISA ECL substrate B (Brown bottle) 6ml 79699 96-well white microplate One

The test method was conducted according to the manual (Assay Protocol) provided by the kit manufacturer, and the test was repeated twice for all samples. To coat the SARS-CoV-2 Spike on the plate, the SARS-CoV-2 Spike was thawed on ice. The thawed reagent contents were distributed appropriately and immediately stored at -80°C. Afterwards, SARS-CoV-2 Spike was diluted with a PBS solution to a concentration of 1㎍/㎖. 50㎕ of the SARS-CoV-2 Spike diluted solution was added to each plate well and left at 4°C for one day. Afterwards, the well plate solution was discarded and washed with 100㎕ of 1x Immno buffer 1 solution. At this time, 1x Immno buffer 1 is 3x Immuno buffer 1 diluted using sterilized distilled water. After washing three times, the plate was tapped on clean paper to remove moisture. Afterwards, 100㎕ of Blocking Buffer 2 was added to the plate well and shaken slowly for 1 hour at room temperature. After this, the supernatant was discarded, and the above washing process was performed again to complete the coating of SARS-CoV-2 Spike on the plate.

20㎕ of the 1x Immuno Buffer was added to each well coated with SARS-CoV-2 Spike, and 10㎕ of the virus inhibition sample solution (Emodin) to be tested was added. At the same time, 10 ㎕ each of positive control (Emodin, Sigma Cat. #324694) and blank were added, and both were left at room temperature for 1 hour. Afterwards, the solution was added to each well according to the composition in Table 6 below.

Blank Positive Control (Emodin) Test Inhibitor 1x Immno Buffer 1 40㎕ 20㎕ 20㎕ Test Inhibitor - - 10㎕ 5% DMSO in water (Inhibitor buffer) 10㎕ 10㎕ - ACE2-His (2.5 ng/㎕) - 20㎕ 20㎕ Total 50㎕ 50㎕ 50㎕

The ACE2-His solution was thawed on ice and then diluted with 1x Immuno buffer 1 to 2.5 ng/㎕. 20㎕ of the diluted ACE2-His solution was added to each well, and reacted by shaking slowly at room temperature for 1 hour. Afterwards, the supernatant was discarded, washed three times with 1x Immno buffer 1 at 100㎕/well, and the plate was tapped on clean paper to remove moisture. Afterwards, 100 ㎕ of Blacking Buffer 2 was added to each well and left at room temperature for 10 minutes. Then, the supernatant was discarded and the wells were washed in the same manner as the washing process above. Afterwards, 20㎕ of the ACE2-His solution diluted in the above process was replaced with 100㎕ of a solution in which Anti-His-HRP was diluted 1000 times with Blocking Buffer 2, and the same process was performed again. Afterwards, 50㎕ of ELISA ECL Substrate A and 50㎕ of ELISA ECL Substrate B were added to the washed wells, and then immediately placed in a reader to measure the fluorescence value during the immune reaction. As the substance in the sample has an antiviral effect, the fluorescence value decreases, and the inhibition rate compared to the blank is calculated and shown in Figures 2 and 3. Figure 2 shows the results for Emodin, the positive control group of the experiment, and Figure 3 shows the antiviral effect of Quillaja high-frequency extract at different concentrations.

Spike RBD of Coronavirus (SARS-CoV-2): As a result of comparing the antiviral activity with Quillaja high-frequency extract using Emodin, known as ACE2 Inhibitor, as a positive control, the inhibition rate of binding between SARS-CoV spike protein and ACE-2 was found to be Emodin showed an inhibition rate of 56%, while Quillaja high-frequency extract at 10% concentration showed an inhibition rate of 99%, and Quillaja high-frequency extract at 100% concentration showed an inhibition rate of 100%. Through these facts, it was confirmed that Quillaja extract has more excellent antiviral efficacy than Emodin, an existing antiviral substance.

Claims (3)

A cosmetic composition for ammonia deodorization containing Quillaia saponaria (Molina) high-frequency water extract,
The Quillaya high-frequency water extract is extracted by adding water to Quillaya and then applying a high frequency of 0.2 to 2 MHz for 1 to 10 minutes at a temperature of 25 to 35 ° C.
The Quillaya high-frequency water extract is a cosmetic composition for ammonia deodorization that removes ammonia molecules. The cosmetic composition for deodorizing ammonia according to claim 1, wherein the content of the Quillaja high-frequency water extract is 0.0001 to 100% by weight. The cosmetic composition for ammonia deodorization according to claim 1, which is in a spray formulation.