KR102488806B1 - A pharmaceutical composition for preventing or treating dry mouth comprising Chamaecyparis obtusa extract - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating xerostomia containing a cypress tree extract.

Description

A pharmaceutical composition for preventing or treating dry mouth comprising Chamaecyparis obtusa extract}

The present invention relates to a pharmaceutical composition for preventing or treating xerostomia containing a cypress tree extract.

Xerostomia is a disease in which the amount of saliva secretion is reduced due to various causes and the oral mucosa becomes dry. It mainly complains of abnormalities in chewing and language functions, and severe bad breath and tooth decay occur, depending on the degree of oral mucosa. Burning sensation in the mouth or ulceration of the oral mucosa will cause severe pain.

Saliva plays an important role in maintaining the oral environment and oral function. That is, saliva has two functions: a food intake function and an oral environment maintenance function. In the food intake function of saliva, saliva maintains the sense of taste through digestion, such as formation of food lumps and action of digestive enzymes, or solubilization of taste stimulants or secretion of gastin (a carbonate dehydrogenase). indicates action. In terms of the oral environment maintenance function, saliva exhibits self-cleaning effects of teeth and mucous membranes, remineralization of teeth, antibacterial action, immune action, tissue repair-promoting action by growth factors, and anti-inflammatory action.

Recently, an increasing number of patients complain of a decrease in salivary secretion caused by various factors. Therefore, social demand for its treatment is increasing. Hypo-salivation accompanies abnormalities in the salivary gland itself caused by radiation therapy or mumps, metabolic diseases (e.g., Basedow's disease, diabetes, etc.), or other diseases such as collagen disease, and also hypo-salivation is caused by stress. It is also caused by factors or side effects of various medicines. With the recent aging of the population, it is thought that the number of patients complaining of decreased salivary secretion as a result of various drug therapies for various complex diseases in which the function of the salivary glands is reduced by aging and that occurs in the elderly will increase in the future. It is becoming.

Hypo-salivation is a result of dry mouth, and the tongue turns red and sometimes cracks, so patients with hypo-salivation complain of pain when eating and difficulty chewing or swallowing. In addition, hyposalivation is known to cause discomfort in the mouth, or taste and speech disorders, and cause denture instability, tooth decay, onset of periodontitis, stomatitis, pneumonia, and digestive dysfunction.

Topical application of artificial saliva is used as a treatment for hyposalivation, but its effect is temporary and limited. Anethol trithione, known as a muscarinic receptor agonist, and cevimelin hydrochloride are used as saliva secretion stimulants, but they are unstable and have problems with the possibility of digestive system side effects such as nausea, vomiting, loss of appetite, and abdominal discomfort. There are some downsides. Under these circumstances, the development of new therapeutic agents for the treatment of hyposalivation is desired.

In order to prevent and treat dry mouth accompanied by severe pain as described above, it is urgently needed to develop a composition for promoting salivary secretion of natural ingredients without side effects other than compounds already known to promote salivary secretion. For example, Korean Patent Publication No. 10-2016-0028423 discloses a composition for preventing and treating xerostomia containing a compound (ID-56D2) isolated from an extract of Squamula japonica as an active ingredient, and Korean Patent Publication No. No. 10-2011-0049015 discloses a composition for enhancing saliva secretion containing Rehmannia glutinosa extract.

Cypress is a member of the Cypress family, and has good wood quality and excellent unique fragrance, so it is very practical. Most of them form a colony in Jeollanam-do, and some of them grow wild in Gyeongsang-do and Jeju-do. Cypress trees are widely known for their antibacterial and sterilizing effects, skin health, insomnia relief, and deodorizing effects.

As a result of research for the treatment of dry mouth, the inventors of the present invention confirmed that the cypress tree extract was effective in treating dry mouth and completed the present invention.

As a result of devising to solve the above problems, the present inventors confirmed that the cypress tree extract has a therapeutic effect on dry mouth, and completed the present invention.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating xerostomia containing a cypress tree extract.

Another object of the present invention is to provide a health functional food composition for preventing or improving dry mouth containing a cypress tree extract.

Another object of the present invention is to provide an oral composition for improving dry mouth containing a cypress tree extract.

The present invention may provide a pharmaceutical composition for preventing or treating xerostomia containing a cypress tree extract.

The cypress tree extract may be a hot water extract.

The dry mouth may be due to at least one selected from the group consisting of Sjogren's syndrome, diabetes, anemia, nutritional deficiency, and aging.

The concentration of the cypress tree extract may be 50 to 500 mg/kg.

The present invention can provide a health functional food composition for preventing or improving dry mouth containing a cypress tree extract.

The present invention can provide an oral composition for improving dry mouth containing a cypress tree (Chamaecyparis obtusa) extract.

The oral composition may be a liquid toothpaste, mouthwash, oral spray or oral ointment.

In experiments using the cypress extract of the present invention, there is an effect of removing methylglyoxal and inhibiting the production of advanced glycation end products (AGEs) derived from methylglyoxal. In addition, as a result of administering cypress tree extract to aging rats, salivary secretion increased in a concentration-dependent manner compared to untreated rats. Therefore, when the cypress tree extract is treated, there is an effect of preventing or treating dry mouth.

Figure 1 measures the amount of methylglyoxal remaining when the cypress tree extract is treated with methylglyoxal.
Figure 2 measures the degree of inhibition of advanced glycation end products produced when bovine serum albumin is treated with methylglyoxal and glucose.
Figure 3 is the result of measuring the amount of saliva secretion when the cypress tree extract was administered to the aging rat.
Figure 4 shows the distribution of blood vessels and serous cells by proceeding with hematoxylin and eosin staining of the salivary glands of rats subjected to aging and administered with cypress extract.
Figure 5 is a TUNEL stain (terminal deoxynucleotidyl transferase (TdT) dUTP nick end labeling) of the salivary glands of mice administered with senescence and cypress tree extract to measure apoptotic cells.

Hereinafter, the present invention will be described in detail.

Xerostomia is a disease in which the amount of saliva secretion is reduced due to various causes and the oral mucosa becomes dry. It mainly complains of abnormalities in chewing and language functions, and severe bad breath and tooth decay occur, depending on the degree of oral mucosa. It is a disease that causes severe pain such as burning sensation in the mouth or ulceration of the oral mucosa. Accordingly, the present inventors confirmed that the cypress tree extract had a therapeutic effect and completed the present invention.

The present invention may provide a pharmaceutical composition for preventing or treating xerostomia containing a cypress tree extract.

The cypress tree extract is effective in removing methylglyoxal related to advanced glycation products (FIG. 1) and has an effect of inhibiting the formation of advanced glycation products (FIG. 2). Furthermore, as a result of administering the cypress extract to aging rats, it was confirmed that salivary secretion increased in a concentration-dependent manner. Through hematoxylin and eosin staining of the salivary glands of rats, a decrease in blood vessels and an increase in serous cells were confirmed in the salivary glands of aged rats administered with cypress extract compared to the salivary glands of aged rats not administered with cypress extract. TUNEL stain Through this, it was confirmed that apoptosis was reduced.

The cypress tree may be, for example, cypress with the scientific name Chamaecyparis obtusa, but is not limited thereto.

Each of the above extraction parts is not particularly limited and all parts can be used, but leaves and stems are preferably used.

The extraction solvent is not particularly limited, and for example, water or an organic solvent may be used, and the organic solvent includes methanol, ethanol, propanol, isopropanol, butanol, etc. Alcohol containing 1 to 4 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclo Various solvents such as cyclohexane may be used alone or in combination, but are not limited thereto.

As an extraction method, any one of methods such as hot water extraction, cold brew extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, and compression may be selected and used, preferably hot water extraction. In addition, the desired extract may be additionally subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation on the preparation method of the extract according to the present invention, and any known method may be used.

As a specific example of hot water extraction, for example, 3 to 20 times the weight of the medicinal material, specifically 5 to 15 times the amount of water added, 80 to 100 ℃, specifically 85 to 95 ℃ for 1 hour to 24 hours , Specifically, it may be extracted for 5 hours to 12 hours, but is not limited thereto.

The extract according to the invention may be subjected to processes such as filtration, concentration, and drying as necessary after the extraction. The method and conditions are not limited, and may be performed by a method known in the art or a condition commonly performed.

The dry mouth may be due to at least one selected from the group consisting of Sjogren's syndrome, aging, diabetes, nutritional deficiency, and anemia, and may preferably be due to aging, but is not limited thereto.

The concentration of the cypress tree extract is not particularly limited, and may be preferably 50 to 500 mg/kg, and more preferably 100 to 250 mg/kg.

The pharmaceutical composition containing the extract according to the present invention is in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions, respectively, according to conventional methods. It can be formulated and used.

Liquid preparations for oral use include suspensions, solutions for oral use, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogeratin and the like may be used.

The amount of the extract according to the present invention may vary depending on the patient's age, sex, and weight, but may be administered in an amount of 01 to 100 mg/kg, preferably 01 to 10 mg/kg once or several times a day. In addition, the dosage may be increased or decreased depending on the route of administration, severity of disease, sex, weight, age, and the like. Therefore, the dosage is not intended to limit the scope of the present invention in any way.

The present invention can provide a health functional food composition for preventing or improving dry mouth containing a cypress tree extract.

The cypress tree extract may be within the above range.

The health functional food of the present invention may be formulated as one selected from the group consisting of tablets, pills, powders, granules, powders, capsules and liquid formulations by further including one or more of carriers, diluents, excipients and additives. Examples of foods to which the extract of the present invention can be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gum, tea, vitamin complexes, health functional foods, and the like.

Additives that may be further included in the present invention include natural carbohydrates, flavors, nutrients, vitamins, minerals (electrolytes), flavors (synthetic flavors, natural flavors, etc.), colorants, fillers (cheese, chocolate, etc.), One or more components selected from the group consisting of pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, antioxidants, glycerin, alcohols, carbonating agents and fruit flesh may be used. .

Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents (thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.

In addition to the above, the health functional food of the present invention is various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like. In addition, the composition according to the present invention may contain fruit flesh for preparing natural fruit juice and vegetable beverages. These components may be used independently or in combination.

Specific examples of the carrier, excipient, diluent, and additive include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium phosphate, calcium Silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate And at least one selected from the group consisting of mineral oil is preferably used.

When formulating the health functional food of the present invention, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

The content of the extract according to the present invention as an active ingredient in the above-described formulation can be appropriately adjusted according to the type and purpose of use, the patient's condition, the type and severity of symptoms, etc., and is preferably 0.001 to 99.9% by weight based on the weight of solid content. May be 0.01 to 50% by weight, but is not limited thereto.

The dosage of the health functional food of the present invention may vary depending on the patient's age, weight, sex, dosage form, state of health and degree of disease, and may be administered once or several times a day at regular time intervals according to the judgment of a doctor or pharmacist. Split doses may also be used. For example, the daily dosage may be 0.5 to 500 mg/kg, preferably 1 to 300 mg/kg, based on the active ingredient content. The above dosage is an example of an average case, and the dosage may be higher or lower depending on individual differences. If the daily dose of the health functional food of the present invention is less than the above dose, a significant effect may not be obtained, and if it exceeds more than that, it is uneconomical and may cause undesirable side effects because it is out of the range of the usual dose. there is.

The present invention may provide an oral composition for improving dry mouth containing a cypress tree (Chamaecyparis obtusa) extract.

The oral composition may be a liquid toothpaste, mouthwash, oral spray or oral ointment.

Hereinafter, examples will be described in detail to explain the present invention in detail.

Example 1. Production of cypress tree extract

The standard extract of Cypress (Chamaecyparis obtusa) was purchased and used from the Korea Plant Extract Bank of the Korea Research Institute of Bioscience and Biotechnology, and the production method of the extract is as follows. After adding 10 times ethanol to each of the leaves, stems or branches of cypress trees collected in the Jeollanam-do region, they were extracted for 3 days, filtered, concentrated under reduced pressure, and freeze-dried.

Example 2. Efficacy test for removing methylglyoxal and inhibiting advanced glycation end products

2-1. Efficacy of removing methylglyoxal

In order to confirm the removal efficiency of methylglyoxal, which is the main cause of final glycation end products, methylglyoxal was treated with methylglyoxal (1 mM, Sigma, MO, USA) at different concentrations to verify the removal efficiency of methylglyoxal. Methylglyoxal was mixed with Cypress extract and incubated with 2N HCI-0.1% 2.4-DNPH at 37°C for 30 minutes. Thereafter, 2.5N NaoH was added to stop color development, and the amount of remaining methylglyoxal was measured by measuring absorbance at 567 nm after 15 minutes of incubation at room temperature. As a result, it was verified that the amount of methylglyoxal remaining in all of the cypress leaf, branch, and stem extracts decreased in a concentration-dependent manner.

2-2 AGEs (Advanced glycation end products) inhibition effect

To confirm the efficacy of inhibiting the production of advanced glycation end products, AGEs, which are one of the main causes of aging, bovine serum albumin (10 mg/ml, Sigma, MO, USA) were mixed together to induce AGEs production, and the cypress extract was treated by concentration, cultured at 37 ° C for 2 to 4 weeks, respectively, and fluorescence was measured. As a result, the production of AGEs was suppressed in a concentration-dependent manner in all extracts of cypress leaves, branches, and stems.

Example 3. Salivary secretion promotion and salivary gland analysis in in vivo aged rats

3-1 Analysis of salivary secretion and salivary glands in rats administered with methylglyoxal

Six-week-old male SD rats (DBL, Korea) were used after acclimatization for one week. Feed and drinking water were provided ad libitum. To induce aging, methylglyoxal (17.25mg/ml, ip) was intraperitoneally administered, and the test drug was administered orally once a day for 2 weeks. The test drug was administered by separating the groups as follows. (1) normal group, (2) aging induction group, (3) cypress tree extract 100 mg/kg administration group, (4) cypress tree extract 250 mg/kg administration group. Autopsies were performed 2 weeks after drug administration for each group. To measure salivary secretion, pilocarpine (1.5 mg/kg) was intraperitoneally injected into rats to induce salivation. Thereafter, the pre-weighed cotton ball was inserted into the oral cavity and left for 20 minutes to absorb the secreted saliva. After 20 minutes, saliva was absorbed and wet cotton balls were collected, weighed, and centrifuged at 10,000 rpm to collect saliva and quantify the amount of saliva. Measurement of salivary secretion was performed 2 weeks after administration. As shown in FIG. 3, it was confirmed that salivary secretion decreased in rats administered with methylglyoxal, and increased in concentration-dependent manner in rats administered with cypress extract.

Analysis of salivary secretion and salivary glands in rats administered with 3-2 D-galactose

Six-week-old male SD rats (DBL, Korea) were used after acclimatization for one week. Feed and drinking water were provided ad libitum. To induce aging, D-galactose (300 mg/ml, ip) was intraperitoneally administered, and the test drug was administered orally once a day for 4 weeks. The test drug was administered by separating the groups as follows. (1) normal group, (2) aging induction group, (3) cypress tree extract 100 mg/kg administration group, (4) cypress tree extract 250 mg/kg administration group. Autopsies were performed 4 weeks after drug administration for each group. To measure salivary secretion, pilocarpine (1.5 mg/kg) was intraperitoneally injected into rats to induce salivation. Thereafter, the pre-weighed cotton ball was inserted into the oral cavity and left for 20 minutes to absorb the secreted saliva. After 20 minutes, saliva was absorbed and wet cotton balls were collected, weighed, and centrifuged at 10,000 rpm to collect saliva and quantify the amount of saliva. Measurement of salivary secretion was performed 2 weeks after administration. As shown in FIG. 3, it was confirmed that the amount of salivary secretion decreased in the rats administered with D-galactose, and the amount of saliva increased in a concentration-dependent manner in the rats administered with the cypress extract.

Example 4. Histopathological analysis of the salivary glands of senescence-induced mice

4-1 Salivary gland analysis through hematoxylin and eosin staining

The salivary glands extracted during autopsy were fixed overnight in 10% neutralized formalin, dehydrated, replaced with xylene three times, and embedded in paraffin. The embedded tissue block was used by making serial sections with a thickness of 4 μm and placing them on slides. Hematoxylin and eosin staining was performed to observe the morphological changes of the salivary glands. The stained slides were observed under an optical microscope. As a result, blood vessels were reduced in the salivary glands of rats administered with the cypress tree extract, compared to the salivary glands of rats without the administration, and an increase in the number of serous cells was confirmed.

4-2 Observation of apoptosis in the salivary glands of senescence-induced mice

The salivary glands extracted during autopsy were fixed overnight in 10% neutralized formalin, dehydrated, replaced with xylene three times, and embedded in paraffin. The embedded tissue block was used by making serial sections with a thickness of 4 μm and placing them on slides. In order to confirm apoptosis of the salivary gland, it was observed through TUNEL staining kit (promega, Wisconsin, United States) staining. As a result, it was confirmed that the number of dead cells in the salivary glands of mice administered with cypress tree extract was smaller than that of mice not administered with cypress tree extract. That is, it was confirmed that the cypress tree extract protects cells from apoptosis due to aging in the salivary gland.

Claims (7)

A pharmaceutical composition for preventing or treating xerostomia, comprising an extract of cypress tree (Chamaecyparis obtusa).
According to claim 1, wherein the cypress tree extract is an ethanol extract, a pharmaceutical composition for preventing or treating xerostomia.
The pharmaceutical composition for preventing or treating dry mouth according to claim 1, wherein the dry mouth is caused by Sjogren's syndrome, diabetes, aging, nutritional deficiency or anemia.
According to claim 1, wherein the concentration of the cypress tree extract is 50 to 500mg / kg, xerostomia prevention or treatment pharmaceutical composition.
A health functional food composition for preventing or improving dry mouth, containing an extract of cypress tree (Chamaecyparis obtusa).
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