A composition for preventing, ameliorating or treating gastritis or peptic ulcer comprising a cinnamomum extract, a fraction of the extract, an isolate of the fraction or a compound isolated therefrom

Abstract

This invention relates to a pharmaceutical or food composition comprising a pretreated extract and isolate of cinnamon exhibiting pharmacological activity, as well as the isolated and purified product. It relates to the active ingredients in cinnamon extracts that exhibit physiological activity.

Description

Technical Field

[0001] This invention relates to a composition capable of preventing, improving, or treating gastritis or peptic ulcers (e.g., gastric ulcers) and exhibiting improved pharmacological or food effects, said composition comprising cinnamon (Cinnamomum cassia) extract, fractions of said extract, isolates of said fractions, or compounds isolated and purified therefrom. Background Technology

[0002] The stomach, part of the digestive tract, is a pouch-like bulge located between the esophagus and the small intestine (duodenum). It stores food that enters through the esophagus, breaks it down for easier digestion, and regulates its delivery to the duodenum, coordinating with the secretion of digestive enzymes for efficient digestion and absorption. Factors that adversely affect human gastrointestinal function are extremely diverse in nature, occurring in the upper, lower, or both digestive tracts, and include a variety of gastrointestinal disease factors, including genetic, physiological, environmental, and psychological factors. Representative diseases of the upper digestive tract include gastritis and peptic ulcers, a general term for both gastric and duodenal ulcers. Gastritis refers to damage and inflammation of the gastric mucosa, while a gastric ulcer refers to damage that penetrates the mucosa and invades the submucosal tissue and muscle layer. Furthermore, a duodenal ulcer is an ulcer that occurs in the duodenum, and gastric and duodenal ulcers are collectively referred to as peptic ulcers. These gastritis and peptic ulcers are known to be caused by factors such as gastric acid imbalance, anti-inflammatory drugs and bacterial infection (called attacking factors) and factors such as mucus, cell regeneration and alkaline secretion (called defensive factors).

[0003] Treatment for gastritis and peptic ulcers primarily includes antacids to neutralize excessive gastric acid secretion, histamine antagonists to inhibit gastric acid secretion, proton pump inhibitors, cholinergic inhibitors, and gastric mucosal protectants that increase the resistance of the gastric lining to digestive juices and aid in recovery. Recently, a pharmacological treatment has been developed that combines the above-mentioned drugs with antibiotics to eradicate Helicobacter pylori. Antacids are characterized by their rapid action, neutralizing gastric acid by raising the pH level in the stomach, thereby protecting the gastric mucosa from acid damage. However, the administration of inorganic substances may affect the smooth muscle of the gastrointestinal tract, leading to constipation, diarrhea, or allergic reactions.

[0004] Cimetidine is a representative histamine receptor-blocking secretion inhibitor widely used for peptic ulcers. Its derivatives include ranitidine, famotidine, and roxatidine. These block histamine receptors and histamine molecules in the gastric mucosa, thus preventing gastric cells from secreting acid. While they have demonstrated excellent anti-ulcer effects in clinical practice, their drawback is that the regenerated mucosa and submucosal tissue after treatment have a weaker structure than normal tissue. Therefore, they are easily damaged again by gastric acid and other attacking factors after drug discontinuation, resulting in a high recurrence rate. Furthermore, ranitidine is ineffective against conditions such as acute gastritis caused by ethanol, thus its gastric mucosal protective ability is poor. Moreover, according to the US Food and Drug Administration, the nitrosamine and dimethylamine groups in ranitidine can decompose and combine over time to produce NMDA (a carcinogen called N-nitrosodimethyl), therefore its use is no longer permitted.

[0005] Recently developed proton pump inhibitors, such as omeprazole and lansoprazole, are known to have a strong inhibitory effect on acid secretion by inhibiting the secretion of acid by parietal cells in the final stage. However, they have a high relapse rate and have been reported to have side effects such as diarrhea, fever, headache, and fatigue.

[0006] In the case of gastric mucosal protectants, long-term treatment is usually required, and the disadvantage is that the dosage is relatively high. However, unlike aggressive inhibitors, the regenerated mucosa is considered to recover to a state similar to normal.

[0007] Helicobacter pylori is a bacterium that resides in the stomach and is known to be a representative cause of recurrent peptic ulcers. It is a Gram-negative bacillus that lives at the junction of gastric mucosal epithelial cells and causes chronic gastric ulcers. While eradication therapy has been successfully implemented, issues remain regarding efficacy, side effects, and the emergence of drug-resistant strains. A safe and reliable eradication method has yet to be established. Summary of the Invention

[0008] Technical issues

[0009] Therefore, the inventors have made great efforts to find a substance that can be used to prevent, improve or treat gastritis and peptic ulcers and exhibit excellent improving pharmacological effects without producing side effects on the human body. Ultimately, they have confirmed that cinnamon extract, fractions of said extract, isolates of said fractions or compounds isolated and purified therefrom can be used to prevent, improve or treat gastritis and peptic ulcers.

[0010] Therefore, the object of the present invention is to provide a pharmaceutical or food composition for the prevention, improvement or treatment of gastritis or peptic ulcers, comprising cinnamon extract, fractions of the extract, isolates of the fractions, an active ingredient isolated therefrom, or a pharmaceutically acceptable salt thereof as the active ingredient.

[0011] Technical solution

[0012] To achieve the above objectives, the present invention provides a pharmaceutical or food composition for the prevention, improvement or treatment of gastritis or peptic ulcers, comprising cinnamon extract, fractions of the extract, isolates of the fractions, and an active ingredient isolated therefrom or a pharmaceutically acceptable salt thereof as the active ingredient.

[0013] As an example, cinnamon extract, fractions of the extract, or isolates of the fraction may include one or more isolated active ingredients selected from the group consisting of compounds represented by the following chemical formulas 1 to 6.

[0014] As an example, the isolated active ingredient can be selected from the group consisting of compounds represented by the following chemical formulas 1 to 6.

[0015] [Chemical Formula 1] Ferulic Acid

[0016]

[0017] [Chemical Formula 2] 4-Hydroxycinnamaldehyde

[0018]

[0019] [Chemical Formula 3] 3-(2-hydroxyphenyl)propionic acid

[0020]

[0021] [Chemical Formula 4] 3,4-Dihydroxybenzaldehyde

[0022]

[0023] [Chemical Formula 5] Syringic acid

[0024]

[0025] [Chemical Formula 6] Vanillic acid

[0026]

[0027] Beneficial effects

[0028] The cinnamon extract, fractions of the extract, or isolates of the fractions of the present invention, as well as the active ingredients of chemical formulas 1 to 6 isolated and purified therefrom, have excellent anti-inflammatory and gastric ulcer-inhibiting effects, and are therefore very suitable as agents for the prevention, improvement, or treatment of gastritis or peptic ulcers, or as functional foods. Attached Figure Description

[0029] Figure 1This is a graph showing the HPLC data of the cinnamon extract contained in the composition of the present invention.

[0030] Figure 2 The graph shows the NO inhibition rate of the cinnamon solvent fraction contained in the composition of the present invention, categorized by type.

[0031] Figure 3 The graph shows the NO inhibition rate of the isolates contained in the compositions of the present invention, categorized by type (N1 to N6).

[0032] Figure 4 The graph shows the NO inhibition rate of the isolates contained in the compositions of the present invention, categorized by type (N2-1 to N2-6).

[0033] Figure 5 The graph shows the NO inhibition rate of the isolates contained in the compositions of the present invention, categorized by type (R2-1 to R2-8).

[0034] Figure 6 This is a schematic diagram of the grading, separation and purification method of cinnamon extract according to the present invention.

[0035] Figure 7 This is a graph comparing the gastric ulcer inhibition rates of cinnamon extract, fractions and active ingredients, ferulic acid, p-cumaraldehyde and 3,4-dihydroxybenzaldehyde contained in the compositions of the present invention with those of Artemisia asiatica extract and rebamipide. Detailed Implementation

[0036] Therefore, the present invention will be described in detail.

[0037] In one aspect, the present invention relates to a pharmaceutical or food composition for the prevention, improvement or treatment of gastritis or peptic ulcers, comprising a cinnamon extract, fractions of the extract, isolates of the fractions, an active ingredient isolated therefrom, or a pharmaceutically acceptable salt thereof as the active ingredient.

[0038] In this invention, "cinnamon" refers to the branches or bark of cinnamon trees that grow to about 8 meters tall in mountainous areas. It is an evergreen broad-leaved tree belonging to the order Ranunculus japonicus and family Cinnamomum camphora. It is native to China and distributed in Sri Lanka, the Indochina Peninsula, and South Korea (Jeju Island).

[0039] In this invention, the term "cinnamon extract" refers to an extract obtained by extracting cinnamon. As a specific example, cinnamon extract can be prepared by drying cinnamon using known conventional methods, or by cutting or crushing it into a suitable size for extraction and extracting it using a suitable extraction solvent, and pretreatment can be performed prior to extraction.

[0040] As an extraction method, known herbal extraction methods can be used, such as hot water extraction, cold water extraction, reflux cooling extraction, or ultrasonic extraction, but are not limited to these. Furthermore, the extract may include the extraction solution itself, a dilution or concentrate of the extraction solution, and a dried substance obtained by drying the extraction solution.

[0041] In one aspect, the cinnamon extract of the present invention may be an extract that has been pretreated before cinnamon is extracted with a polar solvent (i.e., a polar solvent extract of pretreated cinnamon).

[0042] As one aspect, pretreatment can be done by treating cinnamon with a nonpolar solvent.

[0043] As a specific example, a nonpolar solvent could be ethyl acetate.

[0044] The amount of nonpolar solvent used can be 0.5 to 5 times, 0.7 to 4 times, or 1 to 3 times the weight of cinnamon (preferably dry weight), but is not limited thereto.

[0045] As a specific example, pretreatment can be achieved by soaking and stirring chopped or crushed cinnamon in a nonpolar solvent (such as ethyl acetate) for approximately 10 minutes to 5 hours, 20 minutes to 4 hours, or 30 minutes to 3 hours at a temperature of 20°C to 35°C or at room temperature.

[0046] On the one hand, after pretreatment, cinnamon can be washed before extraction with a polar solvent.

[0047] Furthermore, the cinnamon extract according to the present invention can be a polar solvent (preferably water) extract of pretreated cinnamon as described above.

[0048] The amount of polar solvent used for extraction can be 5 to 12 times, 6 to 10 times, or 8 times the weight of cinnamon (preferably dry weight), but is not limited thereto.

[0049] As a specific example, cinnamon pretreated with a nonpolar solvent can be extracted at temperatures of 70°C to 100°C or 80°C to 100°C for 1 to 7 hours, 2 to 6 hours, or 5 hours. Extraction can be performed once or multiple times, once or three times, or once or twice. The pretreated extract can be further filtered, concentrated, and / or dried, and the methods used can be those commonly used in extract preparation, without limitation.

[0050] In one embodiment of the invention, cinnamon is dried and chopped, then ethyl acetate (2 times the volume of cinnamon) is added, and the mixture is soaked and stirred at room temperature for 1 hour. After removing the ethyl acetate, the pretreated cinnamon herbs are washed with water, and then water (8 times the volume of cinnamon herbs) is added. Extraction is carried out at approximately 90°C for 5 hours, and this process is repeated twice. The obtained extract is then filtered, concentrated under reduced pressure, vacuum dried, or spray-dried to prepare a cinnamon ethyl acetate pretreated aqueous extract.

[0051] In this invention, the term "fraction" refers to the result obtained by a fractionation method that separates specific components or groups from a mixture containing various components. The fractionation of the cinnamon extract of this invention can be achieved by fractionating the cinnamon extract using polar solvents (e.g., water, methanol, ethanol, butanol) or non-polar solvents (e.g., n-hexane, ethyl acetate, chloroform) as fractionation solvents to obtain polar solvent fractions and non-polar solvent fractions, respectively. As a specific example, the fractionation of the cinnamon extract can be the ethyl acetate fraction of the cinnamon extract.

[0052] In one aspect, the amount of polar or non-polar solvent (e.g., water, alcohols with a carbon number of 1 (C1) to 4 (C4), chloroform, ethyl acetate, hexane, butanol, or mixtures thereof) added as the fractionating solvent based on 1 kg of cinnamon extract can be extracted and separated 1 to 10 times, preferably 2 to 5 times, to obtain a polar solvent-soluble layer or a non-polar solvent-soluble layer. Furthermore, this solvent fractionation can be performed sequentially.

[0053] In one embodiment of the present invention, the aqueous extract (1 kg) of cinnamon pretreated with ethyl acetate obtained above was fractionated sequentially with H2O / n-hexane, H2O / chloroform, H2O / ethyl acetate, and H2O / n-butanol solvents, 800 ml of each solvent, and then the solvents were removed using a concentrator and a vacuum dryer to prepare cinnamon solvent fractions (n-hexane, chloroform, ethyl acetate, and n-butanol fractions).

[0054] In one instance, fractions of the cinnamon extract showed NO inhibition rates of 4% or more, 7% or more, 19% or more, 50% or more, or 60% or more. In a preferred aspect, the fractions showed NO inhibition rates of 60% to 70%, 60% to 80%, or 65% to 85%.

[0055] In this invention, the term "separate" refers to a substance obtained by further fractionation through conventional separation processes.

[0056] As an example, the isolate can be obtained by various other purification methods, such as passing fractions of the cinnamon extract of the present invention through an ultrafiltration membrane with a constant molecular weight cutoff, or by various chromatographic methods (separating based on size, charge, hydrophobicity, or affinity).

[0057] On the one hand, such a fraction can be obtained by separating fractions with higher physiological activity than the fractions, which are also called active fractions or effective fractions.

[0058] On one hand, the chromatography can be column chromatography, thin-layer chromatography (TLC), or high-performance liquid chromatography (HPLC), but is not limited thereto, and can be performed using various chromatographic methods known in the art. Column chromatography can be used to separate and purify compounds by performing column chromatography using packing materials selected from the group consisting of silica gel, Sephadex, LH-20, ODS gel, C-18 (RP-18), Toyopearl, and XAD resins. If desired, column chromatography can be performed several times by selecting appropriate packing materials, but is not limited thereto. For the use of chromatography, the elution solvent, elution rate, and elution time can be adapted to solvents, rates, or times commonly used in the art.

[0059] In one embodiment of the invention, silica gel (normal phase) and chloroform:methanol were used as elution solvents to elute the ethyl acetate solvent fraction obtained by fractionating cinnamon extract with ethyl acetate, and the separated fractions were concentrated and dried.

[0060] In one instance, the cinnamon extract isolates exhibited NO inhibition rates of 4% or more, 7% or more, 15% or more, 20% or more, 50% or more, or 60% or more. In a preferred aspect, the fractions exhibited NO inhibition rates of 60% to 100%, 70% to 100%, or 75% to 100%.

[0061] In addition, specific active ingredients can be separated and purified from cinnamon isolates using the chromatographic method described above. The isolated and purified active ingredients refer to single substances with physiological activity.

[0062] In one example, chromatography and / or Prep-HPLC (preparative high-performance liquid chromatography) can be used to obtain the isolated and purified active ingredient according to the present invention. Then, a C-18 reversed-phase column can be used as the column for HPLC, and acetonitrile and water can be used as the elution solvent, but are not limited thereto.

[0063] In one instance, the active ingredient isolated and purified according to the invention may be selected from the group consisting of compounds represented by the following chemical formulas 1 to 6.

[0064] [Chemical Formula 1] Ferulic Acid

[0065]

[0066] [Chemical Formula 2] 4-Hydroxycinnamaldehyde

[0067]

[0068] [Chemical Formula 3] 3-(2-hydroxyphenyl)propionic acid

[0069]

[0070] [Chemical Formula 4] 3,4-Dihydroxybenzaldehyde

[0071]

[0072] [Chemical Formula 5] Syringic acid

[0073]

[0074] [Chemical Formula 6] Vanillic acid

[0075]

[0076] The cinnamon extract, fractions of the extract, or isolates of the fractions contained in the compositions of the present invention may include one or more, two or more, three or more, four or more, five or more, six, one to two, one to three, one to four, one to five, one to six, two to three, two to four, two to five, two to six, three to four, three to five, three to six, four to five, four to six, or five to six.

[0077] Furthermore, the compositions of the present invention may include pharmaceutically acceptable salts of isolated and purified active ingredients. As salts, acid addition salts formed from pharmaceutically acceptable free acids are useful. The term "pharmaceutically acceptable salt" as used in this invention refers to any concentration of compound that has an effective effect and is relatively non-toxic and harmless to the patient, and is any organic or inorganic addition salt of the compound wherein the side effects of the salt do not diminish the beneficial therapeutic effect of the compound of the present invention.

[0078] Acid addition salts are prepared by conventional methods, such as dissolving the compound in an excess of aqueous acid and precipitating the salt using a water-miscible organic solvent (e.g., methanol, ethanol, acetone, or acetonitrile). Alternatively, equal molar amounts of the compound and an acid or alcohol (e.g., glycol monomethyl ether) can be heated in water, and the mixture can be evaporated to dryness, or the precipitated salt can be filtered off.

[0079] Then, as free acids, both organic and inorganic acids can be used. As inorganic acids, chloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. can be used. As organic acids, methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutamate, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, etc. can be used, but are not limited to these.

[0080] Furthermore, pharmaceutically acceptable metal salts can be prepared using alkalis. For example, alkali metal or alkaline earth metal salts can be obtained by dissolving the compound in an excess solution of an alkali metal hydroxide or alkaline earth metal hydroxide, filtering the undissolved compound salt, and then evaporating and drying the filtrate. It is then pharmaceutically suitable, but not limited to, to prepare sodium, potassium, or calcium salts as metal salts. Additionally, the corresponding silver salts can be obtained by reacting the alkali metal or alkaline earth metal salts with a suitable silver salt (e.g., silver nitrate).

[0081] Unless otherwise stated, pharmaceutically acceptable salts of the compounds of the present invention include salts of acidic or basic groups that may be present in the compounds described in the above chemical formulas 1 to 6.

[0082] The compositions of the present invention comprise an active ingredient selected from the group consisting of the above chemical formulas 1 to 6, or a cinnamon extract, fractions of the extract, or isolates thereof comprising such an active ingredient, thereby exhibiting excellent effects in preventing, improving, or treating gastritis or peptic ulcers (e.g., gastric ulcers and duodenal ulcers).

[0083] In one instance, the composition of the present invention may be a pharmaceutical composition.

[0084] Based on the total weight of the composition, the pharmaceutical composition of the present invention may contain 10% to 90% by weight of cinnamon extract, fractions of said extract, or isolates of said fraction.

[0085] On the other hand, the pharmaceutical composition may include one or more of the following as active ingredients: ferulic acid, 4-hydroxycinnamaldehyde, 3-(2-hydroxyphenyl)propionic acid, 3,4-dihydroxybenzaldehyde, syringic acid and vanillic acid, respectively represented by chemical formulas 1 to 6.

[0086] When formulating the composition, the content of extracts, fractions, isolates, and active ingredients in the composition can be increased or decreased according to the type of formulation, route of administration, and other requirements.

[0087] The pharmaceutical compositions of the present invention can be administered orally or parenterally and can be used in conventional pharmaceutical formulations. Preferred pharmaceutical formulations include oral administration formulations, such as tablets, pills, powders, granules, hard capsules or soft capsules, liquids, and suspensions. These pharmaceutical formulations can be prepared using pharmaceutically acceptable conventional carriers, such as excipients, binders, disintegrants, glydentants, solubilizers, suspending agents, preservatives, or extenders in the case of oral administration formulations.

[0088] The dosage of the pharmaceutical composition of the present invention can be determined by experts based on various factors such as the patient's condition, age, weight, and disease progression level. However, generally, as an extract, it can be administered once daily or in several separate doses of 9.7 mg to 2919 mg, preferably 29.2 mg to 2919 mg, and more preferably 68.1 mg to 2919 mg. However, if taken long-term, the dosage may be lower than the above range, and the amount of active ingredient used can exceed the above range, as there are no safety concerns.

[0089] Furthermore, the compositions of the present invention can be food compositions. These foods are health supplements, functional foods, or other similar foods, but are not limited to these. The food includes foods in which the cinnamon pretreated extract of the present invention is added to natural foods, processed foods, or common food ingredients. In this document, the term "functional" refers to achieving beneficial effects for health purposes, such as physiological effects that regulate nutrients or modulate human structure and function.

[0090] The health supplements, functional foods, and health products of this invention can be prepared using methods commonly used in the art. During preparation, commonly used raw materials and ingredients can be added. Furthermore, unlike common pharmaceuticals, using currently consumed herbs as raw materials for long-term use does not cause any side effects, and they are highly portable. Therefore, the functional foods of this invention can be taken as adjuvants to enhance the prevention or improvement of gastritis or gastric ulcers. The mixing amount of active ingredients can be appropriately determined according to the intended use (prevention, health care, or treatment). Typically, when preparing food, the content of the fractions or fraction isolates of this invention can be from 10% by weight to 90% by weight.

[0091] On the other hand, the food composition may include one or more of the following as active ingredients: ferulic acid, 4-hydroxycinnamaldehyde, 3-(2-hydroxyphenyl)propionic acid, 3,4-dihydroxybenzaldehyde, syringic acid and vanillic acid, respectively represented by chemical formulas 1 to 6.

[0092] When preparing this composition into food, the content of extracts, fractions, isolates, and active ingredients in the composition can be increased or decreased according to the type of food, route of application, and other requirements.

[0093] The effective dose can be used according to the effective dose of the pharmaceutical composition, but in the case of long-term intake to improve or maintain gastritis or gastric ulcer, it can be lower than the above range, and since cinnamon is usually used as food, there are no safety issues, so its dosage can exceed the above range.

[0094] There are no particular limitations on the type of food. The food composition comprises an active ingredient selected from cinnamon extract, fractions of said extract, isolates of said fraction, or the group consisting of chemical formulas 1 to 6, and can be used as an oral administration formulation, such as tablets, pills, powders, granules, hard capsules or soft capsules, liquids, and suspensions. These formulations may also contain acceptable common food supplements. The term "food supplement" is added to the manufacturing process of the various formulations of the health functional food, and those skilled in the art can appropriately select and use food supplements. Examples of food supplements include various nutrients, vitamins, minerals (electrolytes), flavoring agents (e.g., synthetic and natural flavoring agents), coloring agents and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, ethanol, carbonating agents for carbonated beverages, etc., but the types of food supplements of the present invention are not limited to the above examples.

[0095] Furthermore, the food compositions of the present invention can be added directly or used in conjunction with other foods or food compositions, and can be used appropriately according to common methods. The mixing amount of the active ingredients can be appropriately determined according to their intended use (prevention, improvement, or treatment).

[0096] Examples of food include, but are not limited to, meat, sausage, bread, chocolate, candy, snacks, sweets, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes and other nutritional supplements.

[0097] According to one embodiment of the present invention, the cinnamon extract, fractions of the extract or isolates of the fractions, and active ingredients of chemical formulas 1 to 6 isolated and purified therefrom reduce NO production and inhibit PGE2, thereby exhibiting excellent anti-inflammatory effects and also exhibiting excellent inhibitory effects on gastric ulcers.

[0098] The mode of the present invention

[0099] The invention will be described in more detail below by way of examples. These examples are intended to illustrate the invention only and should not be construed as limiting the scope of the invention to these examples.

[0100] Example 1. Preparation of the extract and solvent fraction of the present invention

[0101] 1) Preparation of cinnamon pretreated extract

[0102] The cinnamon pretreated extract of the present invention was prepared by adding 2 times the amount of ethyl acetate to cinnamon herbs and soaking and stirring at room temperature for at least 1 hour. After removing the ethyl acetate and washing the cinnamon herbs with water, 8 times the amount of water was added, and extraction was carried out at about 90°C for 5 hours (repeated twice). The extract was then filtered, concentrated under reduced pressure, vacuum dried, or spray dried to prepare the cinnamon ethyl acetate pretreated aqueous extract (the resulting extract: 16-26 → 1).

[0103] 2) Preparation of cinnamon solvent fraction

[0104] The cinnamon solvent fraction of the present invention was prepared as follows. Specifically, 1 kg of the cinnamon pretreated extract from Example 1 was sequentially fractionated using 800 ml of each of H2O / n-hexane, H2O / chloroform, H2O / ethyl acetate, and H2O / n-butanol, and then the solvent was removed using a concentrator and a vacuum dryer to prepare the cinnamon solvent fraction.

[0105] [Table 1]

[0106] Yield of cinnamon solvent fraction

[0107] solvent Item obtained (g) Yield (%) Remark n-Hexane 1.3 0.1 - Chloroform 2.6 0.3 - Ethyl acetate 10.2 1.0 active n-Butanol 52.1 5.1 -

[0108] Example 2. Isolation and purification of the bioactive substance of the present invention 1

[0109] 1) Separation of bioactive substances by silica gel column chromatography

[0110] According to the present invention, bioactive substances are separated by silica gel column chromatography as follows. Specifically, the ethyl acetate solvent fraction from the cinnamon solvent fraction of Examples 1 and 2 is used, along with silica gel (normal phase) as the resin of the open column. After packing the silica gel column, the ethyl acetate solvent fraction is separated by adsorption characteristics. The elution solvent is chloroform:methanol = 5:1. After elution, the degree of separation of spots is confirmed by TLC, and the solution is concentrated and dried.

[0111] [Table 2]

[0112] Yield of silica gel column chromatography

[0113] Separate Item obtained (mg) Yield (%) Remark N1 600 6.28 - N2 870 9.10 active N3 1270 13.28 - N4 3020 31.59 - N5 1450 15.17 - N6 2350 24.58 -

[0114] *N=Normal phase

[0115] 2) Separation of bioactive substances by silica gel column chromatography

[0116] According to the present invention, bioactive substances are separated by silica gel column chromatography as follows. Specifically, the isolate N2 from the column chromatography of Example 2-1 and silica gel (normal phase) as an open column resin are used, and after packing the silica gel column, the isolate N2 is separated by adsorption characteristics. The elution solvent is hexane:ethyl acetate = 1:3. After elution, the degree of separation of the spots is confirmed by TLC, and the solution is concentrated and dried.

[0117] [Table 3]

[0118] Yield of silica gel column chromatography

[0119] Separate Item obtained (mg) Yield (%) Remark N2-1 315 43.2 active N2-2 194 26.6 - N2-3 45 6.2 - N2-4 39 5.4 - N2-5 27 3.6 - N2-6 109 15.0 -

[0120] *N=Normal phase

[0121] 3) Purification of single bioactive substances by Prep-HPLC

[0122] According to the present invention, bioactive substances are separated by Prep-HPLC as follows. Specifically, using the silica gel column chromatography isolate N2-1 from Example 2-2 and Prep-HPLC equipped with a C18 column, N2-1 is separated into PH2-1-1 and PH2-1-2 by utilizing the characteristics of the C18 column. The elution solvent is acetonitrile:H2O. After elution, the separation degree of the spots is confirmed on TLC, and the solution is concentrated and dried. PH2-1-1 is purified to compound A by utilizing the characteristics of the C18 column. The elution solvent is acetonitrile:H2O. After elution, the separation degree of the spots is confirmed on TLC, and the solution is concentrated and dried. PH2-1-2 is purified to compound B and compound C by utilizing the characteristics of the C18 column. The elution solvent is acetonitrile:H2O. After elution, the separation degree of the spots is confirmed on TLC, and the solution is concentrated and dried.

[0123] [Table 4]

[0124] Yield of Prep-HPLC purified product

[0125] Separate Item obtained (mg) Yield (%) Notes (purified product) PH2-1-1 313.3 29.5 Compound A PH2-1-2 181.3 17.1 Compound B and Compound C

[0126] *pH = Prep-HPLC

[0127] Example 3. Isolation and purification of the bioactive substances of the present invention 2

[0128] 1) Separation of bioactive substances by C18 column chromatography

[0129] According to the present invention, bioactive substances are separated by C18 column chromatography as follows. Specifically, the separator N2 from the column chromatography of Example 2-1 and C18 resin (reversed phase) as an open column are used, and after packing the C18 column, the separator N2 is separated by adsorption characteristics. The elution solvent is methanol:H2O = 1:1. After elution, the degree of separation of the spots is confirmed on TLC, and the solution is concentrated and dried.

[0130] [Table 5]

[0131] Yield of C18 column chromatography separation

[0132] Separate number Item obtained (mg) Yield (%) Remark R2-1 57 8.5 - R2-2 79 11.7 active R2-3 76 11.3 - R2-4 142 21.0 - R2-5 105 15.6 - R2-6 106 15.8 - R2-7 40 5.9 - R2-8 69 10.2 -

[0133] *R = Inverted

[0134] 2) Purification of single bioactive substances by Prep-HPLC

[0135] According to the present invention, bioactive substances are separated by Prep-HPLC as follows. Specifically, using the isolate R2-2 from the C18 column chromatography of Example 3-1 and Prep-HPLC equipped with a C18 column, the isolate R2-2 is separated into PH2-2-1 and PH2-2-2 by utilizing the characteristics of the C18 column. The elution solvent is acetonitrile:H2O. After elution, the degree of separation of the spots is confirmed on TLC, and the solution is concentrated and dried. PH2-2-1 is purified to compound D by utilizing the characteristics of the C18 column. The elution solvent is acetonitrile:H2O. After elution, the degree of separation of the spots is confirmed on TLC, and the solution is concentrated and dried.

[0136] [Table 6]

[0137] Yield of Prep-HPLC purified product

[0138]

[0139] *pH = Prep-HPLC

[0140] Example 4. Identification of the structure of the purified compound of the present invention by means of physical and chemical properties.

[0141] Measurement of nuclear magnetic resonance (NMR) spectrum

[0142] The NMR spectra were measured by dissolving 10 mg of the purified product in each solvent using 1H-NMR (400 MHz) and 13C-NMR (100 MHz).

[0143] Mass spectrometry measurement

[0144] After separation and purification, a 1 mg dry powder solid sample was measured by chemical analysis using negative ion FAB-mass spectrometry under reduced pressure (10 mmHg). Then, thioglycerol was used as the measurement solvent, and the measurement conditions were an emitter current of 22 eV to 28 eV and an accelerating pressure of 6 kV to 7 kV at the ion source.

[0145] The structural identification results of the purified material obtained by the above method are as follows.

[0146] 1) Compound A

[0147] As a result of the structural identification of compound A, its molecular structure is C. 10 H 10 The molecular weight of O4, as determined by FAB-MS, was 194.18. 1 The H-NMR (CD3OD, 400MHz) spectrum is δ H7.57 (1H,d,J=16.0Hz), 7.17 (1H,d,J=2.0Hz), 7.05 (1H,dd,J=8.0,2.0Hz), 6.80 (1H,d,J=8.0Hz), 6.30 (1H,d,J=16.0Hz), 3.88 (3H,s)ppm, and 13 The C-NMR (CD3OD, 100MHz) spectrum is δ C The concentrations were 170.2, 149.1, 148.0, 144.2, 128.0, 121.6, 116.6, 114.6, 110.3, and 54.4 ppm. Therefore, compound A was identified as ferulic acid.

[0148] 2) Compound B

[0149] As a result of the structural identification of compound B, its molecular structure is C9H8O2, and its molecular weight determined by FAB-MS is 148.16. 1 The H-NMR (CDCl3, 400MHz) spectrum is δ H and 13 The C-NMR (CDCl3, 100MHz) spectrum is δ C The concentrations were 193.9, 161.3, 153.7, 131.6, 127.0, 126.8, and 116.8 ppm, thus compound B was identified as 4-hydroxycinnamaldehyde.

[0150] 3) Compound C

[0151] As a result of the identification of the structure of compound C, its molecular structure is C9H8O3, and its molecular weight determined by FAB-MS is 164.16. 1 The H-NMR spectrum (DMSO-d6, 400MHz) is δ H 7.0 (4H, m), 2.78 (2H, t, J = 7.0 Hz), 2.50 (2H, t, J = 7.0 Hz) ppm, and 13 The C-NMR spectrum (DMSO-d6, 100MHz) is δ C The concentrations were 174.6, 155.6, 130.1, 127.6, 127.3, 119.3, 115.3, 34.1, and 25.9 ppm. Therefore, compound C was identified as 3-(2-hydroxyphenyl)propionic acid.

[0152] 4) Compound D

[0153] As a result of the structural identification of compound D, its molecular structure is C7H6O3, and its molecular weight was determined to be 138.12 by FAB-MS. 1 The H-NMR (CD3OD, 400MHz) spectrum is δ H 9.68 (1H, s), 7.30 (1H, dd, J = 9.48, 1.70 Hz), 7.28 (1H, d, J = 1.41 Hz), 6.90 (1H, d, J = 8.1 Hz) ppm, and 13 The C-NMR (CD3OD, 100MHz) spectrum is δ C The concentrations were 193.8, 154.5, 148.0, 131.6, 127.2, 117.0, and 116.2 ppm, thus compound D was identified as 3,4-dihydroxybenzaldehyde.

[0154] 5) Compound E

[0155] As a result of the structural identification of compound E, its molecular structure is C9H. 10 The molecular weight of O5, as determined by FAB-MS, was 198.17. 1 The H-NMR (CD3OD, 400MHz) spectrum is δ H 7.31 (2H, s) ppm, 3.87 (6H, s) ppm, and 13 The C-NMR (CD3OD, 100MHz) spectrum is δ C The concentrations were 167.2, 147.4, 140.1, 120.6, 106.8, and 55.9 ppm, thus compound E was identified as syringic acid.

[0156] 6) Compound F

[0157] As a result of the structural identification of compound F, its molecular structure is C8H8O4, and its molecular weight was determined to be 168.15 by FAB-MS. 1 The H-NMR (CD3OD, 400MHz) spectrum is δ H 7.55 (2H, m), 6.84 (d, J = 8.8 Hz), 3.89 (3H, s) ppm, and 13 The C-NMR (CD3OD, 100MHz) spectrum is δ C The concentrations were 170.1, 152.6, 148.7, 125.3, 123.2, 115.8, 113.9, and 56.4 ppm, thus compound F was identified as vanillic acid.

[0158] [Measurement of anti-inflammatory effects in Raw 264.7 cells]

[0159] Experimental Example 1. Determination of Nitric Oxide Formation Inhibitory Activity

[0160] Measurement of nitric oxide (NO) involves determining the amount of nitric oxide (NO) in the nitrite and nitrate forms in the cell supernatant. The safe form after oxidation from nitrite to nitrate was measured using Griess reagent (Sigma, USA). Cells were cultured in 2-well plates at a density of 3 × 10⁶ cells / wells. 5 Cells were treated for 24 hours. Within 24 hours, solvent fractions, column isolates, and Prep-HPLC purified products were concentrated and processed into each well. Then, all wells except the normal group were stimulated by adding 100 ng / ml lipopolysaccharide (LPS). Absorbance was measured at 540 nm after 5 minutes of reaction with Gliese's reagent. Calibration curves were prepared using sodium nitrite solution, and nitrite concentration was calculated from absorbance. NO inhibition rate was assessed by comparison with the group treated with LPS only, according to Equation 1 below. Results are shown in Tables 7, 8, 9, 10, 11, and 12 below. Figure 2 , 3 As shown in Figures 4 and 5.

[0161] [Equation 1]

[0162] NO inhibition rate (%) = 100 - [(NO production in the group with added sample × 100) / NO production in the group without added sample]

[0163] *Lipopolysaccharide (LPS) treatment group

[0164] [Table 7]

[0165] Inhibition rate of NO by cinnamon solvent fraction

[0166] solvent NO(μM) NO inhibition rate (%) Remark n-Hexane 57.87 7.1 - Chloroform 49.88 19.9 - Ethyl acetate 20.34 67.4 active n-Butanol 59.59 4.4 -

[0167] [Table 8]

[0168] Inhibition of NO by silica gel column chromatography separations

[0169] Separate NO(μM) NO inhibition rate (%) Remark N1 53.56 16.6 - N2 8.92 86.1 active N3 12.03 81.3 - N4 49.53 22.8 - N5 59.22 7.7 - N6 60.77 5.3 -

[0170] *N=Normal phase

[0171] [Table 9]

[0172] NO inhibition rate of silica gel column chromatography

[0173] Separate NO(μM) NO inhibition rate (%) Remark N2-1 1.54 97 active N2-2 11.57 80 - N2-3 19.93 66 - N2-4 25.21 57 - N2-5 23.40 61 - N2-6 36.23 39 -

[0174] *N=Normal phase

[0175] [Table 10]

[0176] NO inhibition rate of Prep-HPLC purified product

[0177] Separate NO(μM) NO inhibition rate (%) Notes (purified product) PH2-1-1 49.57 0 Compound A PH2-1-2 35.18 29 Compound B PH2-1-2' 48.54 0 Compound C

[0178] *pH = Prep-HPLC

[0179] [Table 11]

[0180] Inhibition rate of NO by C18 column chromatography

[0181] Separate number NO(μM) NO inhibition rate (%) Remark R2-1 37.13 37.8 - R2-2 8.72 85.4 active R2-3 11.38 80.9 - R2-4 40.34 32.4 - R2-5 53.02 11.2 - R2-6 49.15 17.7 - R2-7 40.90 31.5 - R2-8 18.56 68.9 -

[0182] *R = Inverted

[0183] [Table 12]

[0184] NO inhibition rate of Prep-HPLC purified product

[0185] Separate NO(μM) NO inhibition rate (%) Notes (purified product) PH2-2-1 44.41 10 Compound D PH2-2-2 47.83 0 Compound E PH2-2-2' 49.88 0 Compound F

[0186] *pH = Prep-HPLC

[0187] [Measured the inhibitory effect on gastric ulcers in animals]

[0188] Experimental Example 2. Determination of Gastric Ulcer Inhibition Rate

[0189] Assessment of the gastric ulcer index in an animal model of indomethacin-induced gastric ulcer.

[0190] For experimental animals, 7-week-old specific pathogen-free (SPF) male rats were randomly assigned to each group to ensure the most even distribution of average body weight after the acclimatization period (7 days). Administration was performed via single oral doses of Artemisia extract, rebamipide and ethyl acetate-pretreated cinnamon aqueous extract, fraction (N2-1), 3,4-dihydroxybenzaldehyde, ferulic acid, and 4-hydroxycinnamaldehyde, calculated and administered at a dose of 10 mL / kg based on body weight measured on the day of administration. All animals were fasted for 48 hours before administration of the test substances and control drugs, followed by oral administration of each test substance and control drug. Thirty minutes after administration, a pre-prepared indomethacin was administered orally at a dose of 80 mg / kg. Five hours after indomethacin administration, the stomach was removed under diethyl ether anesthesia, and the gastric mucosa was photographed using a digital camera. The area of ​​the damaged portion was analyzed using ImageJ software (NIH, Bethesda, MD). The gastric ulcer index was measured according to the following equation. Results are shown in Table 13 and... Figure 7 As shown.

[0191] [Equation 2]

[0192] Gastric ulcer index (%) = (damaged area / total area) × 100

[0193] [Table 13]

[0194] Group Test material Dosage (mg / kg) Inhibition rate (%) G1 - - - G2 - - - G3 Artemisia extract 20 22.8 G4 Rebapat 30 21.5 G5 Cinnamon extract 24 43.5 G6 Grading (N2-1) 5 74.3 G7 3,4-Dihydroxybenzaldehyde 1.5 61.3 G8 ferulic acid 1.5 61.8 G9 4-Hydroxycinnamaldehyde 2.0 46.1

[0195] like Figure 7 As shown in Table 13, it can be confirmed that cinnamon extract, fractions, and 3,4-dihydroxybenzaldehyde, ferulic acid, and 4-hydroxycinnamaldehyde isolated and purified from them exhibit superior gastric ulcer inhibition rates compared to Stillen, a conventionally available artemisia extract, and rebamipide, a commercially available gastric ulcer treatment. In particular, 4-dihydroxybenzaldehyde, ferulic acid, and 4-hydroxycinnamaldehyde showed very high levels of gastric ulcer inhibition rates even in very low amounts, thus confirming that formulations containing them as active ingredients may be very useful for gastritis and peptic ulcers.

Claims

1. A pharmaceutical composition for preventing, ameliorating or treating gastritis or gastric ulcer, comprising a component selected from the group consisting of a fraction of a cinnamon extract and an isolate of the fraction, the cinnamon extract is a water extract of ethyl acetate-pretreated cinnamon, the fraction is fractionated by using ethyl acetate, and wherein the fraction or the isolate comprises compounds represented by the following Chemical Formulas 1, 2 and 4: [Chemical Formula 1] [Chemical Formula 2] [Chemical Formula 4] 2.A pharmaceutical composition for preventing, ameliorating or treating gastritis or gastric ulcer, comprising a compound represented by the following Chemical Formulas 1, 2 and 4 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] [Chemical Formula 2] [Chemical Formula 4] the pharmaceutical composition further comprises one or more isolated active ingredients selected from the group consisting of compounds represented by the following Chemical Formulas 3, 5 and 6 or pharmaceutically acceptable salts thereof:

3. The pharmaceutical composition of claim 1 or 2, wherein, [Chemical Formula 3] [Chemical Formula 5] [Chemical Formula 6] the isolate is separated by an ultrafiltration membrane or chromatography.

4. The pharmaceutical composition of claim 1, wherein, the chromatography is silica gel column chromatography or column chromatography using C-18.

5. The pharmaceutical composition of claim 4, wherein, 6.The pharmaceutical composition of claim 1 or 2, further comprising a pharmaceutically acceptable carrier. 7.Use of a fraction of a cinnamon extract or an isolate of the fraction in the preparation of a pharmaceutical composition for preventing, ameliorating or treating gastritis or gastric ulcer, the cinnamon extract is a water extract of ethyl acetate-pretreated cinnamon, the fraction is fractionated by using ethyl acetate; and wherein the fraction or the isolate comprises compounds represented by the following Chemical Formulas 1, 2 and 4: [Chemical Formula 1] [Chemical Formula 2] [Chemical Formula 4] 8.Use of a combination of compounds represented by the following Chemical Formulas 1, 2 and 4 in the preparation of a pharmaceutical composition for preventing, ameliorating or treating gastritis or gastric ulcer: [Chemical Formula 1] [Chemical Formula 2] [Chemical Formula 4] ​

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