KR20260043473A - Pharmaceutical composition comprising the extract of sumihyang hot pepper leaves as an effective component for prevention or treatment of thrombosis and health functional food comprising the same - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and a health functional food for the prevention or treatment of thrombosis containing an extract of leaves of Sumihyang, a Korean traditional hot pepper variety ( Capsicum annuum L. ) as an active ingredient. More specifically, the invention relates to a pharmaceutical composition and a health functional food for the prevention or treatment/improvement of thrombosis through inhibition of blood coagulation containing a butanol fraction of an ethanol extract of Sumihyang hot pepper leaves, which has strong prothrombin inhibitory and blood coagulation factor inhibitory activities, as an active ingredient. The extract of leaves of the Korean traditional hot pepper variety Sumihyang ( Capsicum annuum L. ), used as an active ingredient in the pharmaceutical composition and health functional food for the prevention or treatment of thrombosis according to the present invention, exhibits strong antithrombotic activity by inhibiting prothrombin, an enzyme related to thrombus formation, and blood coagulation factors. At the same time, it does not exhibit any hemolytic activity against human red blood cells, has excellent thermal stability, and does not show loss of inhibitory effects on blood coagulation factors or enzymes related to thrombosis formation even under acidic conditions of pH 2 and in plasma. Therefore, it is expected to be usable for the prevention and treatment of thrombosis, such as ischemic stroke and hemorrhagic stroke, through the improvement of blood circulation. Furthermore, since the active ingredient has excellent effects that allow it to be processed into various forms such as extracts, powders, pills, and tablets, and prepared in a form suitable for regular consumption, this invention is highly useful for the pharmaceutical and food industries.

Description

Pharmaceutical composition for the prevention or treatment of thrombosis and health functional food containing Sumihyang hot pepper leaf extract as an active ingredient

The present invention relates to a pharmaceutical composition and a health functional food for the prevention or treatment of thrombosis containing an extract of leaves of Sumihyang, a Korean traditional hot pepper variety ( Capsicum annuum L. ) as an active ingredient. More specifically, the invention relates to a pharmaceutical composition and a health functional food for the prevention or treatment/improvement of thrombosis through inhibition of blood coagulation containing a butanol fraction of an ethanol extract of Sumihyang hot pepper leaves, which has strong prothrombin inhibitory and blood coagulation factor inhibitory activities, as an active ingredient.

As a component of the human body, blood possesses various important functions, including the transport of oxygen, nutrients, and waste products, buffering, maintaining body temperature, regulating osmotic pressure and ion balance, maintaining constant fluid levels, regulating humor, maintaining and regulating blood pressure, and providing biological defense. Normal blood circulation is facilitated by the complementary regulation of the blood coagulation and thrombolytic systems within the body. Among these, the mechanism of the blood coagulation system is reported to involve platelets adhering to and aggregating on the blood vessel walls to form a platelet thrombus, after which the blood coagulation system is activated to form a fibrin thrombus centered around the platelet aggregate.

The formation of fibrin clots involves a multi-step reaction of numerous blood coagulation factors, through which thrombin, which is involved in fibrin coagulation, is activated. This ultimately leads to the generation of fibrin monomers from fibrinogen. These fibrin monomers are polymerized by calcium and bind to platelets and endothelial cells, forming a permanent blood clot by creating a fibrin polymer cross-linked by Factor XIII. Furthermore, thrombin plays a pivotal role in clot formation by activating platelets, Factor V, and Factor VII to promote blood coagulation reactions. Therefore, thrombin inhibitors can be used as highly useful preventive and therapeutic agents for various thrombotic diseases resulting from excessive blood coagulation abnormalities. Meanwhile, the endogenous thrombosis pathway is known to involve the sequential activation of Factor XII, Factor XI, Factor IX, and Factor X, followed by the activation of prothrombin, which ultimately leads to the activation of thrombin; thus, the specific inhibition of blood coagulation factors is also an important target for the development of treatments for thrombotic diseases. To date, various anticoagulants, antiplatelet agents, and thrombolytics such as heparin, coumarin, aspirin, and urokinase have been used for the prevention and treatment of thrombotic diseases; however, their use is limited due to their very high cost, as well as hemorrhagic side effects, gastrointestinal disorders, and hypersensitivity reactions.

Meanwhile, the chili pepper ( Capsicum annuum L.) is an annual plant belonging to the Solanaceae family and is known by various names such as Gocho, Nammancho, Namcho, Dangcho, and Waecho. Its fruits and leaves are edible, and in traditional Korean medicine, the ripe fruit is called Beoncho and is used to treat frostbite, beriberi, and dog bites.

It is known that the origin of the chili pepper is the Amazon River basin in South America, where it was brought to Spain by Columbus in 1493 and spread to Europe, and then to China and Japan around the 17th century. Cultivated chili peppers are native to tropical America, distributed from the southern United States to Argentina, and it is estimated that they were already cultivated quite extensively before the time of Columbus.

Chili peppers have a large number of hybrids resulting from wind-pollinated varieties, and various traditional (Joseon chili) and improved varieties are sold domestically. In Korea, white flowers typically bloom during the summer, and the berry fruits are harvested from July to October; the peppers turn red when ripe. Although there are significant differences depending on the variety, chili peppers generally contain 11–14% moisture, 15–18% crude fat, 5–6% ash, 21–27% crude fiber, 12–15% protein, 25–47% total sugar, and 340 mg/100g of vitamin C. They are also rich in beta-carotene and B vitamins, and the seeds contain a large amount of vitamin E, giving them nutritional excellence beyond being a simple spicy spice. The red color of chili peppers is produced by the interplay of dozens of carotenoid pigments, such as capsanthin, capsorubin, β-carotene, and cryptoxanthin; some of these are converted into Vitamin A after digestion and absorption, serving as a source of Vitamin A for the human body. Furthermore, chili peppers are known to contain a high amount of Vitamin C—twice that of citrus fruits and fifty times that of apples—making them a recognized health food ingredient for boosting immunity, increasing physical strength, and suppressing obesity. In particular, chili peppers contain large amounts of capsaicin, which gives them their characteristic spicy taste. Capsaicin is known to inhibit rancidity and promote the growth of lactic acid bacteria. As a food ingredient enjoyed by those who like spicy flavors, chili peppers in Korea are consumed not only raw but also in fermented foods such as kimchi, gochujang, jangajji, and chogochujang. Additionally, chili leaves are consumed after simmering or blanching, and are also used for pickling.

In 2022, domestic chili pepper production was 68,984 tons, with Gyeongbuk accounting for 17,456 tons, or 25.3% of the national total. However, domestic production and consumption of chili peppers are declining annually. This is attributed to a combination of factors, including the aging of the rural population, labor concentration during cultivation and harvesting, imports of Chinese chili peppers, and climatic influences. Therefore, it is a critical time to develop new chili pepper varieties with superior sensory and functional properties to revitalize the domestic chili pepper industry.

In the case of Cheongyang chili peppers, which are widely consumed in Korea, the multinational agricultural corporation Monsanto holds the ownership of the seeds, resulting in the payment of massive royalties every year. With the recent strengthening of the Nagoya Protocol and the increasing consumer interest in health benefits, it is now necessary to develop domestic seeds and research the excellence of domestic traditional varieties to revitalize the domestic chili pepper industry.

The Yeongyang Chili Pepper Research Institute in Yeongyang-gun, Gyeongbuk, has been continuously conducting research on securing traditional chili pepper varieties to respond to the Nagoya Protocol and secure domestic biodiversity. As of 2022, it has bred more than 26 varieties, restored various varieties such as Subicho, Sumihyang, Yuwolcho, Chilseongcho (Bungeocho), and Goeunbit, and is breeding disease-resistant varieties of traditional chili peppers.

Among domestic traditional chili pepper varieties, Sumihyang was established by cultivating the Ogicho chili pepper, collected in Gyedong-ri, Subi-myeon, Yeongyang-gun in 2000, in an isolated field in Cheonggi-myeon, Yeongyang-gun from 2001 to 2006, selecting individuals with characteristics such as a narrow pedicel, a wrapping calyx shape, and a pointed tip, and cultivating them in a group. After 2007, superior individuals were selected from the group and separated into lines, and then selected through four rounds of self-pollination selection. The finally selected CD-5-7-3-3 was named the line ‘Sumihyang’, and was established by conducting productivity and regional adaptation trials for three years from 2011 to 2013 in Yeongyang-eup (elevation 250m) and Subi-myeon (elevation 400m) in the Yeongyang region.

Sumihyang is a mid-season variety with large fruit and thick skin, resulting in high yield and strong vigor. It has a slightly spicy taste, good gloss, and high sugar content, making it a traditional variety suitable for dried chili peppers due to its excellent dried fruit quality.

Meanwhile, research related to chili peppers has been conducted in various ways, including cultivation, efficacy, and development of processed products. In particular, the antioxidant activity of chili peppers (Byun Eui-baek et al., 2016. Journal of the Korean Society of Food Science and Nutrition 45: 765-770; Kang Hyun-min, 2008. Journal of the Korean Society of Pharmacopuncture and Moxibustion 11: 109-118), antibacterial activity (Seo Hwa-jung, 1999. Journal of the Korean Society of Food Science and Nutrition 28: 94-99), cancer cell growth inhibitory activity (Jung Mi-ra et al., 2011. Journal of the Korean Society of Food Science and Nutrition 40: 642-648), and the antioxidant, antibacterial, and tyrosinase inhibitory activities of chili pepper leaves (Kim Ji-hye et al., 2003. Journal of the Korean Society of Food Preservation and Distribution 10: 540-546) are known. Recently, improved varieties such as Dangjo peppers are being sold commercially, and it has been reported that Dangjo peppers contain five times more α-glucosidase inhibitor (AGI), which prevents diabetes, than regular peppers, and that Dangjo pepper leaves contain more than five times more AGI than the fruit. Studies related to the antidiabetic activity of chili peppers include a study on line selection for antidiabetic activity (Cho Myung-cheol et al., 2007. Journal of Biological Environmental Control 233-239), lipid metabolism and effects of Dangjo chili peppers in a diabetes animal model (Lee Seung-beom, 2023. Master's thesis, Chung-Ang University), a study on the antidiabetic effects of Dangjo chili pepper leaves in mice (Ju Ah-shin, 2015. Master's thesis, Wonkwang University), the effect of capsaicin diet on the metabolism of diabetic rats (Jeon Seong-hyun, 2011. Master's thesis, Chonnam National University), metabolite and genome-wide association analysis of alpha-glucosidase inhibitors in chili pepper leaves (Shamuel Tillahun, 2022. Ph.D. thesis, Jeonbuk National University), and the molecular mechanism of capsaicin-induced anti-obesity and anti-metastatic effects (Lee Gong-rak, 2014. Ph.D. thesis, Ulsan University).

Studies on Sumihyang include only “Development of the high-yield traditional chili pepper variety Sumihyang” (Hwang Ji-eun et al., 2015. Presentation in the Korean Society of Horticultural Sciences 135), and there are currently no studies related to the useful physiological activities of Sumihyang.

Studies related to the antithrombotic activity of chili peppers include the investigation of thrombolytic ability, immunogenicity, and cytotoxic effects of strains isolated from chili peppers (Seo et al., 2007. Korean Journal of Food Science and Technology 39: 315-322), the identification and functional study of strains isolated during chili pepper fermentation (Seo, 2006. Master's thesis, Korea University), and the search for thrombolytic enzyme-producing strains and the investigation of optimal conditions for enzyme production (Choi, 2003. Korean Journal of Food Science and Nutrition 32: 976-980), but these are reports related to the thrombolysis of strains involved in chili pepper fermentation.

Emran T.B. et al. reported that chloroform extracts of Bangladeshi medicinal plants (Trema orientalis L., Bacopa monnieri L., Capsicum frutescens L., Brassica oleracea L. and Urena sinuata L.) have thrombolytic activity (Emran TB. et al., 2015. Effects of organic extracts and their different fractions of five Bangladeshi plants on in vitro thrombolysis. BMC Complement Altern Med. Apr 23;15:128. doi: 10.1186/s12906-015-0643-2), but these are chloroform extracts of plants different from chili peppers, and are known for their thrombolytic activity that dissolves formed blood clots. Therefore, to date, there have been no studies reporting that the chili pepper itself inhibits blood clot formation and exhibits strong antithrombotic activity, and in particular, no studies on the strong antithrombotic activity of the traditional chili pepper variety Sumihyang are known.

Most patents related to chili peppers involve the development of processed products such as gochujang and cheonggukjang containing chili peppers, as well as patents for eco-friendly antimicrobial microorganisms capable of controlling diseases such as chili pepper blight. Regarding the beneficial activities of chili peppers, Korean registered patents disclosed include No. 10-2298042 [Antioxidant and anti-inflammatory composition containing chili pepper seed extract], No. 10-1899917 [Antioxidant composition containing chili pepper extract as an active ingredient and method of manufacturing the same], No. 10-0487945 [Chili pepper leaf extract having immune-enhancing activity and method of manufacturing the same], No. 10-2205201 [Composition for removing senile odor and nonenal containing chili pepper extract], No. 10-1834834 [Method of manufacturing functional soap containing chili pepper or paprika extract], and No. 10-1539688 [Cosmetic composition effective for improving skin photoaging containing chili pepper capsicum resin and method of manufacturing the same], as well as Korean published patents Disclosure No. 10-2013-0051717 [Anticancer active chili seed extract and functional food containing the same], Disclosure No. 10-2022-0100260 [Effective extraction method of a chili extract high in polyamine having anti-inflammatory activity], etc.

Regarding patents related to the antithrombotic activity of chili peppers, Korean registered patent No. 10-0810532 discloses [Bacillus bacteria having thrombolytic ability, immune cell activation ability and cytotoxic activity and gochujang containing the same], and Korean published patent No. 10-2021-0103788 discloses [a composition for preventing thrombosis and improving blood circulation] containing a mixed extract of yuzu, stevia, balloon flower root, Cheongyang chili pepper, ginger, and radish, but no patents regarding the antithrombotic activity of chili peppers are known.

In addition, regarding chili peppers, patents related to antidiabetic activity are most concentrated. Patents related to the antidiabetic activity of chili peppers include [Oil-roasted chili leaf tea extract having antidiabetic activity] in Patent No. 10-1237694, [Korean confectionery containing antidiabetic chili leaves as an active ingredient and method of manufacturing the same] in Patent No. 10-1412068, [Food composition for improving diabetes and pharmaceutical composition for preventing or treating diabetes containing a new variety of chili pepper (deposit number KCTC18413P) or its extract] in Patent No. 10-1793256, [Fermented chili leaf vinegar and method of manufacturing the same] in Patent No. 10-1356611, [Method of manufacturing a functional beverage using chili peppers and the beverage] in Patent No. 10-0689870, and [Effervescent granular tea composition containing Dangjo chili pepper extract and method of manufacturing the same] in Patent No. 10-2015-0075151. In addition, [Gochujang for blood sugar control in diabetic patients] is disclosed in Public Patent No. 10-2011-0014418, [Method for manufacturing gochujang with anti-diabetic efficacy] in Public Patent No. 10-2015-0067857, and [Anti-diabetic and anti-obesity fermented composition containing fermented chili peppers and fermented chili pepper pomace] fermented with selected lactic acid bacteria in Public Patent No. 10-2020-0047939. Furthermore, regarding Korean traditional chili peppers, [Composition for skin whitening containing Subicho chili pepper seed extract as an active ingredient] is known in Public Patent No. 10-2018-0108981.

Meanwhile, only the young shoots of chili pepper leaves are used as a vegetable, and the rest are discarded, making their utilization extremely limited. Furthermore, research data regarding the components, efficacy evaluation, processing, and storage of chili pepper leaves is severely lacking. Currently, dried chili pepper leaves are the most common product related to chili pepper leaves. Recently, beverages containing Salicho chili pepper leaves and Jerusalem artichoke juice, known for their excellent anti-diabetic activity, are being sold. Additionally, chili pepper leaf beverages and pill products containing Wonki No. 2, known for its excellent anti-diabetic activity, have also been developed.

Studies related to chili pepper leaves to date include the antioxidant, antibacterial, and tyrosinase inhibitory activities of solvent fractions of chili pepper leaves (Kim Ji-hye et al., Journal of the Korean Society of Food Preservation and Distribution 2003, 10, 540-546), the antioxidant and cancer cell proliferation inhibitory effects of chili pepper leaf extracts (Jeon Geon-wook et al., Journal of the Korean Society of Food Science and Nutrition 2008, 37, 1079-1083), antioxidant activity and antioxidant components by chili pepper leaf variety (Koo Kang-mo et al., Journal of Applied Biological Chemistry 2009, 52, 70-76), quinone reductase activity and functional components by chili pepper leaf variety (Kang Young-hwa et al., 2007 Autumn Conference of the Korean Society of Plant Resources, 2007, 368.2), phenolic compounds of chili pepper leaves (Lee, Jae-Woong et al., 2008 Autumn Conference of the Korean Society of Medicinal Crops, 2008, 412), and studies using different pretreatment methods Studies on the physicochemical properties and antioxidant capacity of chili pepper leaf extracts (Hong, Seong-Wook, 2015, Master's thesis, Hoseo University), isolation of phenolic compounds with radical scavenging activity from chili pepper leaves (Choi, Jang-Ki et al., Journal of the Korean Society of Pharmacognosy, 2007, 38, 258-262), isolation and characterization of complement system-activating polysaccharides from chili pepper leaf hot water extracts (Na, Kyung-Soo et al., Journal of the Korean Society of Life Sciences, 2002, 12, 87-95), isolation and identification of secondary metabolites from chili pepper leaves (Cho, Jin-Kyung, 2009, Master's thesis, Kyung Hee University) and reporting of hACAT-1 inhibitory activity (human-derived Acetyl-CoA acetyltransferase) of isolated substances, isolation of chili pepper leaf (leaf of Capsicum annuum L.) extract and search for α-glucosidase inhibitory active substances using an automated HPLC/SPE/HPLC system (Sepboxsystem) (Kim, Min-Sun et al., Journal of the Korean Society of Applied Biological Chemistry). Antioxidant, antidiabetic, and anticancer activities of alpha-glucosidase inhibitors in chili pepper leaves have been reported, such as in *2021. 64, 25-32* and metabolite and whole-genome association analysis (Samuel Tillahun, 2022, Ph.D. dissertation, Jeonbuk National University). Additionally, regarding studies on light stress in chili pepper leaves, the photosynthetic response and photoprotection mechanism of chili pepper leaves to light stress have been reported (Lee et al., Journal of the Korean Society of Environmental Agriculture, 2006. 25, 109-117), and regarding studies on pesticide residue characteristics in chili pepper leaves, pesticide residue characteristics in chili peppers and chili pepper leaves according to cultivation environment, variety, and processing method have been reported (Lee et al., Journal of the Korean Society of Pesticide Science, 2006. 10, 99-106). However, to date, the antithrombotic activity of the butanol fraction of Sumihyang chili pepper leaf extract has not been reported.

To date, patents related to chili pepper leaves include: Korean Registered Patent No. 10-1714392 [Method for manufacturing noodles with added chili pepper leaf powder and noodles with added chili pepper leaf powder manufactured by said method]; No. 10-1483938 [Method for manufacturing Wonki No. 1 chili pepper leaf kimchi and Wonki No. 1 chili pepper leaf kimchi manufactured by said method]; No. 10-1548592 [Method for manufacturing chili pepper leaf tea containing Salvia miltiorrhiza]; No. 10-2133804 [Method for manufacturing seasoning sauce for chicken using chili pepper leaf powder]; No. 10-2042151 [Composition for improving intestinal flora containing chili pepper leaf extract]; No. 10-2066845 [Plasma-treated chili pepper leaf extract with enhanced anti-inflammatory effect and method for manufacturing the same]; and No. 10-2016994 [Chili pepper leaf extract with enhanced skin whitening effect and method for manufacturing the same]. Korean Patent Publication No. 10-2034178 [Plasma-treated chili pepper leaf extract with enhanced skin whitening effect and method for preparing the same], No. 10-1963630 [Anti-inflammatory composition containing chili pepper leaf extract obtained through enzyme treatment as an active ingredient], No. 10-2233744 [Composition for improving, preventing, or treating leaky gut syndrome containing chili pepper leaf extract], No. 10-2018643 [Skin whitening composition containing chili pepper leaf extract obtained through enzyme treatment and ultra-high pressure homogenization process as an active ingredient], and No. 10-2416786 [Composition for improving, preventing, or treating obesity and metabolic diseases containing chili pepper leaf extract] are disclosed, and Korean Patent Publication No. 10-2020-0082637 [Fermented vinegar of chili pepper leaf extract having anti-obesity effect], and No. 10-2022-0041735 [New variety Although [a composition for lowering blood sugar containing chili pepper leaf extract as an active ingredient], [a composition for the prevention, improvement, or treatment of diabetes containing chili pepper leaf extract and Jerusalem artichoke extract as active ingredients] in Patent No. 10-2024-0015980, [a composition for the improvement, prevention, or treatment of obesity and metabolic diseases containing chili pepper leaf extract] in Patent No. 10-2020-0125156, and [a composition for the prevention, improvement, or treatment of metabolic diseases containing a new variety of chili pepper leaf extract as an active ingredient] in Patent No. 10-2022-0132349 have been disclosed, the antithrombotic activity of the butanol fraction of the Sumihyang chili pepper leaf extract has not been known to date.

KR 1020220132349 A

Koo Kang-mo et al., Journal of applied biological chemistry 2009, 52, 70-76

The present invention was devised to solve the problems of the prior art as described above. The problem to be solved by the present invention is to provide a pharmaceutical composition and a health functional food for the prevention or treatment/improvement of thrombosis through inhibition of blood coagulation, containing an extract of the leaves of the Korean traditional hot pepper variety Sumihyang (Leaves of Sumihyang, Korean traditional hot pepper, Capsicum annuum L.) as an active ingredient.

In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention or treatment of thrombosis containing, as an active ingredient, an extract of the leaves of the Sumihyang variety (Leaves of Sumihyang, Korean traditional hot pepper, Capsicum annuum L. ), registered under Korean Plant Variety Protection No. 7248.

It is preferable that the above chili pepper leaf extract is a butanol fraction derived from an ethanol extract.

In addition, the present invention provides a blood coagulation inhibitor containing, as an active ingredient, an extract of the leaves of the Sumihyang variety, which is registered under Korean Plant Variety Protection No. 7248.

It is preferable that the above chili pepper leaf extract is a butanol fraction derived from an ethanol extract.

In addition, the present invention provides a health functional food for preventing or improving thrombosis containing, as an active ingredient, an extract of the leaves of the Sumihyang variety, registered under Korean Plant Variety Protection No. 7248.

It is preferable that the above chili pepper leaf extract is a butanol fraction derived from an ethanol extract.

The extract of leaves of the Korean traditional hot pepper variety Sumihyang ( Capsicum annuum L. ), used as an active ingredient in the pharmaceutical composition and health functional food for the prevention or treatment of thrombosis according to the present invention, exhibits strong antithrombotic activity by inhibiting prothrombin, an enzyme related to thrombus formation, and blood coagulation factors. At the same time, it does not exhibit any hemolytic activity against human red blood cells, has excellent thermal stability, and does not show loss of inhibitory effects on blood coagulation factors or enzymes related to thrombosis formation even under acidic conditions of pH 2 and in plasma. Therefore, it is expected to be usable for the prevention and treatment of thrombosis, such as ischemic stroke and hemorrhagic stroke, through the improvement of blood circulation. Furthermore, since the active ingredient has excellent effects that allow it to be processed into various forms such as extracts, powders, pills, and tablets, and prepared in a form suitable for regular consumption, this invention is highly useful for the pharmaceutical and food industries.

Figure 1 is a photograph of the mature fresh fruit, dried fruit, and fruit set status of Sumihyang.
Figure 2 shows the whole plant powder of Sumihyang, Cheongyang, and Baekduhong chili pepper leaves from the left.

The present invention will be described in detail below.

The inventors of the present invention, in order to test the antithrombotic efficacy of the leaves of the Korean native variety of Sumihyang chili pepper, prepared an extract of Sumihyang chili pepper leaves by a specific method and prepared sequential organic solvent fractions thereof, and evaluated their antithrombotic and antioxidant activities. They recovered the butanol fraction of the ethanol extract of the leaves of the Korean native variety of Sumihyang chili pepper as an antithrombotic active component. By confirming that the butanol fraction exhibits no hemolytic activity against human red blood cells while possessing excellent thermal and acid stability, they intended to utilize it as a pharmaceutical composition and health functional food for the prevention or treatment/improvement of thrombosis.

The chili pepper variety "Sumihyang" of the present invention was registered as Korean Plant Variety Protection No. 7248 on June 8, 2018. In addition, it was registered as Korean Variety Name Registration No. 02-0004-2693 on March 20, 2015, under the variety name "Sumihyang".

Specifically, in order to develop pharmaceutical compositions and health functional foods for the prevention or treatment/improvement of thrombosis using chili pepper leaves, which are known in folk medicine to be effective for various diseases of the vascular and circulatory systems, digestive systems, and metabolic systems, the inventors prepared various solvent extracts and their sequential organic solvent fractions using the leaves of the Korean native variety Sumihyang chili pepper. They evaluated the antithrombotic activity of these extracts and fractions by measuring direct thrombin inhibition (Thrombin Time: TT), prothrombin inhibition (Prothrombin Time: PT), and activated partial thromboplastin time (aPTT) against human thrombin, and confirmed that the butanol fraction of the ethanol extract of the Korean native variety Sumihyang chili pepper leaves exhibited very strong anticoagulant activity. Furthermore, they confirmed that the fractions did not exhibit hemolytic activity against human red blood cells, unlike other Sumihyang fractions, thereby confirming their practical applicability.

Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of thrombosis containing an extract of leaves of Sumihyang, Korean traditional hot pepper ( Capsicum annuum L. ) as an active ingredient.

It is preferable that the above-mentioned Korean traditional variety Sumihyang chili pepper leaf extract is a butanol fraction of an ethanol extract of a mature Korean traditional variety Sumihyang chili pepper leaf.

As a preferred embodiment, the chili pepper leaf extract of the present invention may be a butanol fraction obtained by sequentially fractionating hexene, ethyl acetate, and butanol after ethanol extraction.

In addition, the present invention provides a blood coagulation inhibitor containing an extract of leaves of Sumihyang, Korean traditional hot pepper ( Capsicum annuum L. ) as an active ingredient.

It is preferable that the above-mentioned Korean traditional variety Sumihyang chili pepper leaf extract is a butanol fraction of an ethanol extract of a mature Korean traditional variety Sumihyang chili pepper leaf.

As a preferred embodiment, the chili pepper leaf extract of the present invention may be a butanol fraction obtained by sequentially fractionating hexene, ethyl acetate, and butanol after ethanol extraction.

In addition, the present invention provides a health functional food for preventing or improving thrombosis containing an extract of leaves of Sumihyang, Korean traditional hot pepper ( Capsicum annuum L. ) as an active ingredient.

It is preferable that the above-mentioned Korean traditional variety Sumihyang chili pepper leaf extract is a butanol fraction of an ethanol extract of a mature Korean traditional variety Sumihyang chili pepper leaf.

As a preferred embodiment, the chili pepper leaf extract of the present invention may be a butanol fraction obtained by sequentially fractionating hexene, ethyl acetate, and butanol after ethanol extraction.

Below, the method for preparing the Korean traditional variety Sumihyang chili pepper leaf extract and efficacy experiments of the present invention will be explained in more detail.

The present invention comprises the steps of: extracting leaves of the Korean native variety Sumihyang chili pepper with various solvents and preparing sequential organic solvent fractions thereof; evaluating the antithrombotic activity of the extracts and fractions; and investigating the stability of the butanol fraction of the Sumihyang chili pepper leaf ethanol extract.

The "Korean traditional variety Sumihyang chili pepper leaf extract" included in the composition of the present invention can be obtained by the steps of extracting mature Korean traditional variety Sumihyang chili pepper leaves with an organic solvent, filtering the extract using a filter mesh of 0.06 mm or less, and concentrating it under reduced pressure.

The organic solvent used in the present invention may be water (cold water, hot water), hexene, methylene chloride, acetone, ethanol, anhydrous or aqueous lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, ethanol, propanol, butanol, etc.), a mixed solvent of the lower alcohol and water, etc., and hot water or 95% ethanol extraction is most preferable.

The above ethanol extract can be further obtained by sequentially or separately fractionating it with organic solvents of hexene, ethyl acetate, and butanol to obtain a hexene fraction, an ethyl acetate fraction, a butanol fraction, and a water residue.

As a preferred embodiment, the Korean traditional variety Sumihyang chili pepper leaf extract of the present invention may use the butanol fraction of a mature Sumihyang chili pepper leaf ethanol extract.

In the present invention, an ethanol extract of the leaves of the Korean native variety Sumihyang chili pepper and its fractions were prepared, and when these were adjusted to a concentration of 5 mg/ml and the thrombin time, prothrombin time, and APT time were measured, it was confirmed that the butanol fraction of the ethanol extract exhibited stronger inhibition than aspirin, thereby inhibiting the formation of blood clots. Furthermore, unlike other fractions, the butanol fraction of the Sumihyang ethanol extract showed no hemolytic activity against human red blood cells, suggesting that it can replace existing antithrombotic agents such as aspirin, which have a high risk of side effects.

The Korean traditional variety Sumihyang chili pepper leaf extract of the present invention can be prepared into a powder through conventional pulverization processes such as vacuum drying, freeze-drying, or spray drying. These are not degraded by various degrading enzymes in plasma and maintain their activity even under heat treatment at 100°C and at a pH of 2 in the human stomach.

The active ingredient of the present invention can be used for the prevention or treatment of various diseases related to thrombosis. The diseases include, for example, arterial thrombosis such as acute myocardial infarction, chest pain, shortness of breath, loss of consciousness, ischemic stroke, hemorrhagic stroke, headache, motor abnormalities, sensory abnormalities, personality changes, visual impairment, epileptic seizures, pulmonary thrombosis, deep vein thrombosis, lower extremity edema, pain, and acute peripheral arterial occlusion, and venous thrombosis such as deep vein thrombosis, portal vein thrombosis, acute renal vein occlusion, cerebral sinus thrombosis, and central retinal vein occlusion.

A pharmaceutical composition containing the active ingredient of the present invention can be formulated into various forms according to conventional methods to suit each intended use, such as oral formulations like powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, and injectable formulations like sterile injectable solutions, and can be administered orally or through various routes including intravenous, intraperitoneal, subcutaneous, rectal, and local administration.

These pharmaceutical compositions may additionally include carriers, excipients, or diluents, and examples of suitable carriers, excipients, or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Additionally, the pharmaceutical compositions of the present invention may additionally include fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc.

As a preferred embodiment, solid formulations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid formulations are formulated by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc., with the above pharmaceutical composition. In addition, in addition to simple excipients, lubricants such as magnesium stearate, talc, etc. may be used.

As a preferred embodiment, oral liquid formulations may be exemplified as suspensions, liquid formulations, emulsions, syrups, etc., and may include various excipients in addition to commonly used simple diluents such as water and liquid paraffin, such as humectants, sweeteners, flavorings, preservatives, etc.

As a preferred embodiment, formulations for parenteral administration may be exemplified as sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized agents, suppositories, etc. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. Injectables may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, etc.

The active ingredient of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level may be determined based on factors including the type and severity of the patient's disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as a single or multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects, taking all of the above-mentioned factors into consideration, and this can be easily determined by a person skilled in the art.

As a preferred embodiment, the effective amount of the active ingredient in the pharmaceutical composition of the present invention may vary depending on the patient's age, gender, and body weight, and generally, 1 to 5,000 mg, preferably 100 to 3,000 mg per kg of body weight, may be administered daily or every other day, or divided into 1 to 3 doses per day. However, since the dosage may be increased or decreased depending on the route of administration, severity of the disease, gender, body weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention may be administered to a subject via various routes. Any mode of administration may be anticipated, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intradural, or intracerebroventricular injection.

In the present invention, "administration" means providing a specific substance to a patient by any appropriate method, and the route of administration of the pharmaceutical composition of the present invention may be oral or parenteral through any general route capable of reaching the target tissue. Additionally, the composition of the present invention may be administered using any device capable of delivering the active ingredient to target cells.

In the present invention, "object" includes, but is not specifically limited to, humans, monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, or guinea pigs, and preferably means mammals, more preferably humans.

In addition, the health functional food of the present invention can be used in various ways in foods and beverages, etc., that are effective in preventing or improving thrombosis. Foods containing the active ingredients of the present invention include, for example, various types of food, beverages, chewing gum, tea, vitamin complexes, health supplements, etc., and can be used in the form of powder, granules, tablets, capsules, or beverages.

The active ingredient of the present invention can generally be added in an amount of 0.01 to 15% by weight of the total food weight, and the health drink composition can be added in a ratio of 0.02 to 10g, preferably 0.3 to 1g, based on 100ml.

In addition to containing the above compound as an essential component in the indicated proportions, the health functional food of the present invention may contain food-grade acceptable food additives, such as natural carbohydrates and various flavoring agents, as additional components.

Examples of the above natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, as well as common sugars and sugar alcohols such as xylitol, sorbitol, and erythritol.

As the above flavoring agents, natural flavoring agents such as taumatin; rebaudioside A or glycyrrhizin, such as stevia, and synthetic flavoring agents such as saccharin and aspartame may be used. The ratio of the above natural carbohydrates is generally about 1 to 20g, preferably about 5 to 12g, per 100ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention may contain various nutritional supplements, vitamins, minerals, flavoring agents such as synthetic and natural flavoring agents, coloring agents and thickening agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. Furthermore, the health functional food of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, vegetable beverages, etc. These ingredients may be used independently or in combination. The proportion of such additives is generally selected in the range of 0.01 to about 20 parts by weight per 100 parts by weight of the active ingredient of the present invention.

The present invention will be explained in more detail below through examples. The following examples are merely preferred embodiments of the present invention, and the scope of the present invention is not limited to the scope of the following examples.

[Example]

Example 1: Preparation of Korean traditional Sumihyang chili pepper leaf powder and color difference of the powder

Mature Sumihyang chili pepper leaves cultivated in a rain-sheltered greenhouse at the Yeongyang Chili Pepper Research Institute in Gyeongbuk in 2023 were harvested, dried at 50°C for one day, and ground to produce Sumihyang chili pepper leaf powder (Figs. 1 and 2). The average particle size of the prepared powder was 0.5–2.0 mm, and the color difference was 59.37, showing a color difference similar to that of conventional improved varieties Cheongyang and Baekduhong chili pepper leaf powders (Table 1, Fig. 2).

[Table 1] Color difference of Sumihyang chili pepper leaf powder

Example 2: Preparation and component analysis of Sumihyang chili pepper leaf ethanol extract and its sequential organic solvent fractions

An ethanol extract was prepared using the Sumihyang chili pepper leaf powder from Example 1. Specifically, 10 times the amount of 95% ethanol was added to the prepared sample, and the extract was extracted twice at room temperature. The extract was collected, filtered, and then concentrated under reduced pressure to produce a powder, thereby preparing the ethanol extract. The prepared ethanol extract was suspended in water and subjected to sequential organic solvent fractionation with hexene, ethyl acetate, and butanol, and finally, the water residue was recovered. The fractionation efficiency and polyphenol content analysis results are shown in Table 2.

[Table 2] Analysis of Yield and Polyphenol Content of Sumihyang Pepper Leaf Ethanol Extract and Its Fractions

The yield of the ethanol extract of the leaves of the Korean native variety Sumihyang chili pepper was 13.7%, and the yield of the butanol fraction was 35.77%. This means that approximately 4.9g of butanol extract can be recovered from 100g of Sumihyang chili pepper leaf powder. The total polyphenol content of the ethanol extract of the Korean native variety Sumihyang chili pepper leaves was 28.49mg/g, which was similar to 31.3mg/g of the Sumihyang chili pepper extract. Among the sequential organic solvent fractions, the polyphenol content of the butanol fraction was 26.5mg/g, notably showing the highest value of 33.64mg/g in the water residue. Therefore, it was expected that the butanol fraction of the Sumihyang chili pepper leaf extract would exhibit various useful physiological activities. At this time, the total polyphenol content was analyzed by adding 50 μl of Folin-Ciocalteau and 100 μl of _{saturated Na₂CO₃} solution to 400 μl of the extract solution, leaving it at room temperature for 1 hour, and measuring the absorbance at 725 nm. Tannic acid was used as the standard reagent.

Example 3: Evaluation of the antithrombotic activity of Sumihyang chili pepper leaf extract and its fractions

The blood coagulation inhibitory activity of the ethanol extract of the leaves of the Korean native variety Sumihyang pepper and its fractions from Example 2 was evaluated, and the results are shown in Table 3. The blood coagulation inhibitory activity was evaluated according to previously reported methods (Sohn et al., 2004. Kor. J. Pharmacogn 35. 52-61; Kwon et al., 2004. J. Life Science, 14. 509-513; Ryu et al., 2010. J. Life Science, 20. 922-928), and thrombin time, prothrombin time, and APT time were measured. Commercially available control plasma (MD Pacific Technology Co., Ltd, Huayuan Industrial Area, China) was used, and the thrombin time, prothrombin time, and APT time measurements were performed as follows.

Thrombin Time

At 37°C, 50 μl of 0.5 U thrombin (Sigma Co., USA), 50 μl _of 20 mM CaCl₂, and 10 μl of sample extracts of various concentrations were mixed in the tube of an Amelung coagulometer KC-1A (Japan) and reacted for 2 minutes. Afterward, 100 μl of plasma was added, and the time until the plasma coagulated was measured. Aspirin (Sigma Co., USA) was used as a control, and DMSO was used as the solvent control instead of the sample. In the case of DMSO, a coagulation time of 32.1 seconds was observed. The thrombin inhibitory effect was expressed as the average of experiments repeated at least three times, and thrombin inhibitory activity was expressed as the value obtained by dividing the coagulation time upon sample addition by the coagulation time of the solvent control.

Prothrombin time

70 μl of standard plasma (MD Pacific Co., China) and 10 μl of sample solutions of various concentrations were added to the tubes of an Amelung coagulometer KC-1A (Japan). After heating at 37°C for 3 minutes, 130 μl of PT reagent was added, and the time until the plasma coagulated was recorded as the average of three repeated experiments. Aspirin (Sigma Co., USA) was used as a control, and DMSO was used as the solvent control instead of the sample. In the case of DMSO, a coagulation time of 18.1 seconds was observed. Prothrombin inhibitory activity was expressed as the value obtained by dividing the coagulation time upon sample addition by the coagulation time of the solvent control.

aPTT (activated Partial Thromboplastin Time)

100 μl of plasma and 10 μl of sample extracts of various concentrations were added to the tube of an Amelung coagulometer KC-1A (Japan) and heated at 37°C for 3 minutes. Then, 50 μl of aPTT reagent (Sigma, ALEXIN ^™ ) was added, and the mixture was incubated again at 37°C for 3 minutes. Afterward, 50 μl of _CaCl₂ (35 mM) was added, and the time until the plasma coagulated was measured. DMSO was used as the solvent control instead of the sample, in which case a coagulation time of 55.1 seconds was observed. The aPTT result was expressed as the average of three repeated experiments, and blood coagulation factor inhibitory activity was expressed as the value obtained by dividing the aPTT time at the time of sample addition by the aPTT time of the solvent control.

[Table 3] Inhibitory activity of Sumihyang chili pepper leaf extract and its fractions

As shown in Table 3, when aspirin (antithrombotic agent trade name: Protect), used as a control, was measured at a concentration of 1.5 mg/ml, the thrombin time, prothrombin time, and APT time were extended by 1.37, 1.31, and 1.56 times, respectively, confirming that it exhibited excellent antithrombotic activity. On the other hand, when the ethanol extract of Sumihyang pepper was measured at a concentration of 5 mg/ml, the thrombin time, prothrombin time, and APT time increased by 1.11, 1.23, and 1.12 times, respectively, compared to the control group, showing relatively weak activity compared to aspirin. However, in the case of the butanol fraction of the Sumihyang ethanol extract, the thrombin time, prothrombin time, and APT time increased by 1.24, 2.45, and 1.31 times, respectively, compared to the control group at a concentration of 5 mg/ml, demonstrating excellent anticoagulant activity. Therefore, it was determined that the butanol fraction of the ethanol extract of the leaves of the Korean native variety Sumihyang chili pepper can effectively inhibit blood clot formation by inhibiting various blood coagulation factors and prothrombin, and can replace aspirin, which is known to cause side effects such as gastrointestinal disorders.

Example 4: Evaluation of the Antioxidant Activity of Korean Traditional Sumihyang Pepper Leaf Extract and Its Fractions

Since polyphenol components of natural products are closely related to antioxidant activity and oxidative stress is also closely related to thrombus formation (Wang, L. et al., Sci. Rep. 2017, 7, 12429; Martinez, M. et al., Free Radic. Biol. Med. 2013, 65, 411-418; Bijak, M. et al., Thromb. Res. 2012, 130, e123-e128), the antioxidant activity of the leaf extract of the Korean native variety Sumihyang chili pepper and its fractions was evaluated, and the results are shown in Table 4. At this time, the extract was dissolved in DMSO (dimethylsulfoxide) and diluted to an appropriate concentration to be used for measuring DPPH (1,1-diphenyl-2-picryl hydrazyl) anion radical scavenging activity and ABTS [2,2-azobis (3-ethylbenzo thiazoline-6-sulfonate)] cation radical scavenging activity. First, for DPPH scavenging activity, 380 μl of 2 x ^10⁻⁴ M DPPH solution dissolved in 99.5% ethanol was added to 20 μl of samples diluted to various concentrations, mixed, and reacted at 37°C for 30 minutes. Subsequently, the absorbance was measured at 516 nm using a microplate reader (Asys Hitech, Expert96, Asys Co., Austria). DPPH radical scavenging activity was expressed as a percentage of the sample-added group versus the non-added group. For ABTS scavenging activity, 5 ml of 7 mM ABTS (Sigma Co., USA) and 88 ml of 140 mM potassium persulfate were mixed and the mixture was kept at room temperature in a light-blocked environment for 16 hours to form ABTS cations. Subsequently, this solution was diluted with ethanol until the absorbance value at 414 nm was 1.5. 190 ml of the prepared diluted solution was mixed with 10 ml of samples prepared at various concentrations, reacted at room temperature for 6 minutes, and the absorbance was measured at 734 nm. The ABTS radical scavenging activity was then calculated using the following formula.

Meanwhile, for the measurement of nitrite scavenging activity, the sample solution was added to a nitrite solution (1 mM), and 0.1 N HCl was added to adjust the pH to 1.2. After reacting at 37°C for 1 hour, Griess reagent (Sigma Co., USA) was added and mixed. Subsequently, the amount of residual nitrite was determined by measuring the absorbance at 520 nm after leaving the solution at room temperature for 15 minutes. The nitrite scavenging activity (%) was calculated using the following formula.

For the evaluation of reducing power, a modified method of Oyaizu et al. (Ahn et al., 2011. J. Life Sci. 21: 576-583) was used. To 2.5 ml of the sample dissolved in ethanol, 2.5 ml of 0.2 M sodium phosphate buffer (pH 6.6) and 2.5 ml of 10% potassium ferricyanide were added and reacted at 50°C for 20 minutes. Afterward, 2.5 ml of 10% trichloroacetic acid was added to terminate the reaction, and the supernatant was collected by centrifuging at 4000 rpm for 10 minutes. The collected supernatant was diluted twofold with distilled water, mixed with a freshly prepared 0.1% ferric chloride solution at a ratio of 5:1 (v/v), and evaluated by measuring the absorbance at 700 nm.

[Table 4] Antioxidant activity of Sumihyang chili pepper leaf ethanol extract and its fractions

As shown in Table 4, vitamin C used as a control exhibited very strong antioxidant activity at a concentration of 0.5 mg/ml, showing 88.9% DPPH anion scavenging activity, 90.4% ABTS cation scavenging activity, 65.3% nitrite scavenging activity, and a reducing power of 1.631. The ethanol extract of the leaves of the Korean native variety Sumihyang chili pepper showed excellent antioxidant activity at a concentration of 0.5 mg/ml, exhibiting 43.07% DPPH anion scavenging activity, 73.63% ABTS cation scavenging activity, 59.49% nitrite scavenging activity, and a reducing power of 0.410. Meanwhile, the butanol fraction of the ethanol extract of Sumihyang chili pepper leaves exhibited excellent antioxidant activity, showing 49.07% DPPH anion scavenging activity, 36.43% ABTS cation scavenging activity, 33.16% nitrite scavenging activity, and 0.425 reducing power at a concentration of 0.5 mg/ml. Therefore, it is judged that the strong antioxidant activity of the butanol fraction of Sumihyang chili pepper leaf extract contributes to antithrombotic activity.

Example 5: Human erythrocyte hemolytic activity of Sumihyang chili pepper leaf extract and its fractions

To evaluate the potential for acute toxicity of the leaf extract of the Korean native variety Sumihyang chili pepper and its fractions, human erythrocyte hemolytic activity was evaluated. The hemolytic activity was evaluated according to a previous report (Son Ho-yong, 2014 · Korean J. Microbiol. Biotechnol. 42: 285-292). Simply put, 100 μl of human erythrocytes washed three times with PBS were placed in a 96-well microplate, 100 μl of sample solutions of various concentrations were added, and the mixture was reacted at 37°C for 30 minutes. Afterward, the reaction mixture was centrifuged (1,500 rpm) for 10 minutes, and 100 μl of the supernatant was transferred to a new microtiter plate, after which the degree of hemoglobin leakage due to hemolysis was measured at 414 nm. DMSO (2%) was used as the solvent control for the samples, and Triton X-100 (1 mg/ml) was used as the experimental control for erythrocyte hemolysis. Hemolytic activity was calculated using the following formula.

[Table 5] Human erythrocyte hemolytic activity of Sumihyang chili pepper leaf extract and its fractions

As shown in Table 5, DMSO and water used as controls showed no hemolytic activity, and it was confirmed that Triton X-100 lysed 100% of red blood cells at a concentration of 1 mg/ml. In addition, it was confirmed that amphotericin B, which is used as an anticancer and antifungal agent, lysed more than 50% of red blood cells at a concentration of 0.025 mg/ml.

Meanwhile, it was confirmed that the ethanol extract of Sumihyang chili pepper leaves and its ethyl acetate fraction, butanol fraction, and water residue of the present invention did not exhibit erythrocyte hemolysis up to a concentration of 1 mg/ml, indicating no acute toxicity or erythrocyte hemolytic activity. On the other hand, the hexene fraction of the Sumihyang chili pepper ethanol extract showed 25.12% hemolytic activity at a concentration of 1 mg/ml. These results suggest that the butanol fraction of the Sumihyang chili pepper ethanol extract of the present invention can safely replace antithrombotic agents that cause side effects, such as aspirin, in the future without exhibiting separate hemolytic activity.

Example 6: Evaluation of plasma, acid, and thermal stability of the butanol fraction of Sumihyang chili pepper leaf ethanol extract

The plasma stability, thermal stability, and acid stability regarding antithrombotic activity were confirmed for the butanol fraction of the Sumihyang chili pepper leaf ethanol extract obtained in Example 2 above. The butanol fraction did not show a decrease in blood coagulation inhibitory activity even after heat treatment at 100°C for 1 hour, treatment at pH 2 (0.01M HCl) for 1 hour, and treatment in plasma for 1 hour. Therefore, it was confirmed that the butanol fraction of the Sumihyang chili pepper leaf contains an antithrombotic active substance with acid resistance and heat resistance, and thus confirmed that there is a high possibility of practical use as an antithrombotic food or pharmaceutical material in the future.

[Research and Development Project]

- Project ID: PJ0169582023

- Supervising Department: Research Policy Division

- Specialized research management agency: Rural Development Administration

- Research Project Name: Development of Regionally Specialized Crop Technology

- Research Project Title: Development of Technology for High-Quality Stable Production and Value-Added Enhancement to Expand Consumption of Regionally Specialized Traditional Chili Peppers

- Contribution rate: 100%

- Research period:

- Total research period: April 1, 2021 – December 31, 2024 (33 months)

- Multi-year agreement research period: April 1, 2021 – December 31, 2024 (33 months)

- Research period for the current year: 2024.01.01.–2024.12.31.(12 months)

- Supervising Organization: Yeongyang Chili Pepper Research Institute, Gyeongsangbuk-do Agricultural Research and Extension Services

- Commissioned Research Institution: Andong National University Industry-Academic Cooperation Foundation

- Actual research implementing agencies: Yeongyang Chili Pepper Research Institute, Gyeongsangbuk-do Agricultural Research and Extension Services, Industry-Academic Cooperation Foundation, Andong National University

Claims (4)

A pharmaceutical composition for the prevention or treatment of thrombosis containing, as an active ingredient, an extract of the leaves of the Sumihyang chili pepper variety, Korean Plant Variety Protection No. 7248. A pharmaceutical composition for the prevention or treatment of thrombosis according to claim 1, characterized in that the extract is a butanol fraction after ethanol extraction. A blood coagulation inhibitor comprising the active ingredient described in claim 1 or 2. A health functional food for the prevention or improvement of thrombosis containing the active ingredient described in Paragraph 1 or 2.