KR102229760B1 - Fraction of Melissa Leaf Extract and Novel Pharmaceutical Composition Comprising the Same - Google Patents

Abstract

The present invention relates to a Melissa officinalis leaf extract fraction and a novel pharmaceutical composition comprising the same. In the case of a pharmaceutical composition for the prevention or treatment of obesity comprising the Melissa officinalis leaf extract fraction according to the present invention as an active component, it is possible to prevent or treat obesity by exhibiting significant weight suppression, blood triglyceride suppression, blood sugar suppression, and total cholesterol suppression effects.

Description

TECHNICAL FIELD [Fraction of Melissa Leaf Extract and Novel Pharmaceutical Composition Comprising the Same}

The present invention relates to a novel pharmaceutical composition comprising the extract fraction of melissa leaf and the same as an active ingredient.

Melissa officinalis is a perennial herb belonging to the Lamiaceae family ( Labiatae ), and has the nickname of Lemon Balm.

The extract of melissa leaf contains flavonoids, triterpene acids, volatile oils, glycosides such as alcohol and phenolic compounds, and caffeic acid derivatives. In particular, the flavonoids contained in large amounts in Melissa leaf are cinnaroside, cosmosin, rhamnocitrin, isoquercitrin, etc., and ursolic acid is contained as terpenic acid. have. It contains derivatives of hydroxycinnamic acid such as rosmaric acid, which is one of the nonvolatile components that have recently attracted attention, and volatile oils include geraniol, neal, and sheet. It contains citronellal and eugenol.

Obesity is a condition that can cause various health problems due to excessive accumulation of fat in the body due to an imbalance between energy intake and consumption. The cause is mostly due to lifestyles such as excessive nutritional intake and lack of physical activity, and in rare cases, it can be secondary to medication or disease.

According to statistics from the World Health Organization (WHO) 2014, worldwide 1.9 billion of adults over the age of 18 are overweight, of which 600 million have reached obesity. Obesity is one of the main factors that increase the incidence of cardiovascular disease, type 2 diabetes, and several types of cancer. More than 2.8 million people die each year from being overweight or obese.

As the living standards of modern people improve, obesity has become a global social problem. According to the 2015 National Health and Nutrition Survey, Korean people's fat intake has increased by 5.9g over the past 10 years, and the prevalence of obesity has increased by 1.9% compared to 2005. In particular, the prevalence of obesity in men increased significantly to 39.7%, and the prevalence of hypercholesterolemia increased by 9.9% to 17.9%. As the living standards of modern people improve, obesity has become a global social problem.

Various drugs have been developed to improve obesity, and drugs are largely divided into appetite suppressants and liposuction suppressants according to their mechanism of action. However, drugs such as phentermine and diethylpropion, which suppress appetite, have side effects such as elevated blood pressure, dizziness, headache, tremor, and dry mouth. Absorption of dissolved vitamins is inhibited, and there are fat stools, fat excretion, frequent bowel movements, and stool incontinence.

Therefore, there is a need to develop safe drugs with few side effects in order to prevent or treat obesity.

Nonalcoholic hepatic steatosis refers to an excessive accumulation of triglycerides in the liver regardless of alcohol consumption. Among healthy and lean people, when the triglyceride content in the liver is higher than 95% or the triglyceride particle ratio in the cytoplasm is 5% or more in the liver cells, it is defined as simple fatty liver among non-alcoholic fatty liver diseases. Non-alcoholic fatty liver is reported to be 10-24% in the general population and 58-74% in obese people.

Nonalcoholic steatohepatitis (NASH) is a disease in which fat accumulates in the liver, causing hepatocytes to balloon and inflame, followed by fibrosis, resulting in cirrhosis, and in some cases, liver cancer.

The number of patients with non-alcoholic steatohepatitis is rapidly increasing, but the pathogenesis of the disease is not clear and various causes are acting on it, making it difficult to treat by controlling one mechanism or cause, and there is no approved treatment.

Since non-alcoholic steatohepatitis progresses slowly and chronically, there are generally no specific symptoms, but liver functions such as an increase in the activity of alkaline phosphatase (ALP) or aminotransferase in the blood, which are a measure of liver function. It gets worse gradually.

If left untreated, non-alcoholic steatohepatitis progresses to fibrosis or hardening of the liver, resulting in poor prognosis. Therefore, in order to block the progression of non-alcoholic steatohepatitis to cirrhosis, it is necessary to suppress the accumulation of fat in the liver and prevent further inflammation and fibrosis, but no treatment plan has been suggested. Therefore, there is a need to develop an effective therapeutic agent that can treat non-alcoholic steatohepatitis.

The macula lutea is a part of the retina of the eye that receives light most clearly and accurately because the eye cells are dense, and a disease that causes vision impairment due to various causes of this macular degeneration is called macular degeneration. do. The macular degeneration is one of the three major causes of blindness along with glaucoma and diabetic retinopathy. The biggest cause of macular degeneration is age increase, and family history, race, and smoking are also known causes.When the macula is damaged, the ability to recognize details such as small prints, facial features, or small objects in the eyes is lost. It is done.

There are two types of macular degeneration: non-exudative (dry) macular degeneration and exudative (wet) macular degeneration, and 90% of those with macular degeneration have dry symptoms. In dry macular degeneration, waste products can form yellow deposits called drusen, which can accumulate in the tissues below the macula. The presence of drusen interferes with blood flow to the retina, especially the macula, and when blood flow decreases, it reduces the supply of nutrients to the macula, stopping or atrophy of the efficient action of photosensitive cells. In the case of wet macular degeneration, new weak blood vessels can grow in or below the retina, causing fluid and blood to leak into the space below the macula.

This macular degeneration is considered a common disease in the elderly, but recently, it is known that patients in their 4's and 50's are increasing rapidly, and as a major cause of lowering the age group onset of macular degeneration, westernization of diet, such as an increase in fat intake, has been pointed out.

Lutein is a carotenoid that is naturally produced and does not have vitamin A activity and exists in a natural state. It is contained in large amounts in green and yellow vegetables. Lutein, one of the carotenoids most frequently present in food and human blood, is known to play an antioxidant effect in plants and to protect plants from ultraviolet rays.In humans, it is a major component of the macula and lens of the eye. It is known as an ingredient that plays an important role in improving health and vision.

Recently, however, a research team at the Moran Eye Hospital affiliated with Utah State University found a round, yellowish crystal-like substance in the center and inside of the retina of a woman who took excessive lutein for a long time through the JAMA Ophthalmology. Reported as being done. In the case of the patient, it was found that he took a lutein supplement of 20 mg every day for 8 years and steadily consumed spinach, broccoli, kale, and avocado rich in lutein. Therefore, the research team estimated that lutein was precipitated in the eyeball, crystals were formed, and macular degeneration occurred.

Therefore, although lutein has an effect of improving visual acuity in macular degeneration, long-term or excessive ingestion rather causes side effects that cause macular degeneration. Therefore, it is necessary to develop a method to prevent, alleviate, or inhibit this.

Psoriasis is a chronic inflammatory skin disease in which the skin of the whole body is repeatedly covered with silvery-white scales, reddish papules or plaques of varying sizes, and is characterized by proliferation of the epidermis and inflammation of the dermis. And appears at a frequency of 1 to 2% of the population. Psoriasis is a chronic skin disease that occurs when a small millet-like rash occurs on the skin, and dead skin cells such as white dandruff are piled up over the affected area. So, if it spreads a lot, almost all of the skin on the whole body is covered with the rash. Through this process, psoriasis progresses chronically, sometimes getting better on its own, and in many cases, it spreads throughout the body.

In psoriasis, the skin cells that make up dead skin cells rapidly increase, causing layers of dead skin such as dandruff to build up on the skin. The cause of psoriasis is not yet clearly identified, but it is reported as an immune disorder. In addition, genetic factors, environmental factors, drugs, skin irritation, dryness, upper respiratory tract inflammation, and mental stress are factors that cause or worsen psoriasis. It is being discussed.

To treat psoriasis, vitamin D derivative ointment, narrow-band UVB treatment, bath photochemotherapy (bath-PUVA), cyclosporine, and the like are used. The causal treatment method for psoriasis is not known yet. This is because the pathological cause of the outbreak has not been accurately identified. Treatment methods such as vitamin D derivative ointment, narrow-band UVB treatment, bath-PUVA, and cyclosporine are only temporary symptom relief methods and recur. It can't be called a method.

Cancer is one of the most common causes of death worldwide. About 10 million new cases occur every year, accounting for about 12% of all causes of death, making it the third leading cause of death.

Among cancers, breast cancer is the most common malignant tumor that causes more than 40,000 deaths in women every year, and early diagnosis is very important.However, despite the treatment of many known anticancer drugs, the survival rate when the cancer progresses or metastases. This is a situation that has not improved.

Chemotherapy, a typical anticancer therapy, is currently being used as the most efficient treatment for cancer, alone or in combination with other treatments such as radiotherapy. However, the efficacy of a cancer treatment drug in chemotherapy depends on its ability to kill cancer cells, but there is a problem that it can act on not only cancer cells but also general cells when the drug is used, and thus drugs for efficiently treating cancer are required.

As the duration of diabetes increases, various complications occur throughout the body. Diabetic retinopathy is a chronic complication that is directly referenced in the diagnosis of diabetes. In diabetic patients, more than 60% within 10 years of diabetes diagnosis, within 20 years. It appears in more than 90%.

Diabetic retinopathy is well known to be more specific to hyperglycemia than other chronic diabetes-related complications.

Diabetic retinopathy can be classified into early non-proliferative diabetic retinopathy (NPDR) and late proliferative diabetic retinopathy (PDR) according to the degree of progression. In addition, when accompanied by macular edema (DME, diabetic macular edema), severe visual acuity may decrease even at this time.

If such diabetic retinopathy is detected early, the progression and worsening of retinopathy can be prevented with appropriate management, but if the condition is not properly managed, severe vision loss or blindness may occur. Currently, diabetic retinopathy is considered to be a major cause of blindness in adults, and despite this clinical significance, the development of drugs for preventing or treating diabetic retinopathy has not been actively made.

Therefore, the need for drug development for preventing or treating diabetic retinopathy has been raised.

Glaucoma is a disease in which the optic nerve that transmits visual information to the brain is disturbed, resulting in atrophy of the optic nerve and narrowing the field of vision. As types of glaucoma, primary open-angle glaucoma (POAG), normal tension glaucoma, primary angle-closure glaucoma, developmental glaucoma, secondary glaucoma, and the like are known. In addition, ocular hypertension, in which the visual field is normal, but the intraocular pressure is chronically high, is one of the risk factors for glaucoma.

In the treatment of glaucoma, lowering the intraocular pressure and preventing further visual field disturbances are considered the top priority, and to lower the intraocular pressure, medication, laser treatment, and surgery are performed, but these conventional treatments are not satisfactory for all patients. There is a need for a glaucoma treatment drug containing an active ingredient with a new mechanism of action or a new structure, which is not present in the existing treatment drugs.

Sjogren's syndrome is a chronic disease that causes secretion disorders due to the invasion of lymphocytes into the exocrine glands. Characteristic symptoms are dry eye and dry mouth caused by lymphocytic infiltrates of the lacrimal and salivary glands. With the loss of tears and saliva, characteristic changes in the eyeball (referred to as aqueous tear deficiency or dry keratitis conjunctivitis) and characteristic changes in the oral cavity (this leads to tooth damage, increased oral infections, difficulty swallowing, and mouth dryness). Pain) can be triggered. The patient may also have joint inflammation (arthritis), muscle inflammation (myositis), nerve inflammation (neuropathy), thyroid inflammation (thyroiditis), kidney inflammation (nephritis), lung inflammation or inflammation of other parts of the body, or lymph nodes Swelling can develop. In addition, the patient may experience fatigue and sleep disturbances. Sjogren syndrome mainly affects middle-aged women.

Currently, there is no known treatment for Sjogren's syndrome, and there is no effective treatment to restore glandular secretion. Existing treatments are generally symptomatic and supportive, and include hydration therapy (e.g., to relieve symptoms of dry eyes and mouth) and various forms of lubrication. Prescription drugs are available, including cyclosporine, which is helpful in treating chronic dry eye, and sebimeline or pilocarpine, which is helpful in stimulation of salivary fluid flow. Anti-inflammatory agents such as methotrexate and hydroxychloroquine have also been prescribed for improvement of musculoskeletal symptoms. However, none of the drugs currently available are ideal due to the wide range of serious side effects.

Atherosclerosis, also called atherosclerosis, is a disease in which arterial wall thickening or tissue degeneration occurs and hardens. As the inner diameter of the artery becomes thicker and the inner diameter narrows, a disorder appears in the blood flow that supplies oxygen and various nutrients, thereby causing various diseases. Atherosclerosis is likely to occur in the cerebral or coronary arteries. In the case of cerebral atherosclerosis, it is known to cause headache, dizziness, and mental disorder and cause encephalopathy, and in the case of coronary atherosclerosis, it is known to cause pain and arrhythmia in the heart, causing angina pectoris and myocardial infarction. In addition, arteriosclerosis is deeply related to the development and progression of diseases such as metabolic syndrome or lipid dysemia such as obesity (overweight), blood clots, hypercoagulation and thrombosis-inducing conditions (arteries and veins), including central obesity, and diabetic Causes or worsens chronic complications.

In addition, when the concentration of triglycerides in the blood is increased due to obesity, hyperlipidemia, etc., the rate at which macrophages are converted into foam cells or killed by triglycerides is increased. That is, as macrophages die, their immune function and triglyceride metabolism function decrease, and the function of removing formed foam cells decreases, thereby increasing the likelihood of developing or worsening atherosclerosis. Therefore, in order to prevent the onset or worsening of arteriosclerosis, it is important to keep macrophages functioning to remove foam cells even when the blood fat concentration is high, or to minimize their death by triglycerides.

Currently, statin drugs such as rosuvastatin and simvastatin are mainly used for the treatment of arteriosclerosis, but this is known to have a side effect of reducing the activity of macrophages that remove foam cells. In addition, statin drugs have been pointed out as a problem in causing side effects such as diabetes, muscle dystrophy, hyperglycemia, and the like, and research on natural products with fewer side effects is required.

Arthritis is largely divided into rheumatoid arthritis and osteoarthritis (degenerative arthritis). Rheumatoid arthritis is a chronic, inflammatory, multisystem disorder that typically involves joints. Symptoms include pain and swelling in various joints such as fingers, hands, feet, wrists, ankles, and knees, and cause abnormalities in various organs such as muscles, skin, lungs, and eyes. The cause of rheumatoid arthritis is caused by genetics, infection by pathogens, and abnormal immune responses, but it is not known precisely. The pathogenesis is interleukin-1 (IL-1), tumor necrosis factor (TNF), prostaglandin E2 (PGE2), collagenase, proteinase, substance-P, etc., which are released from immune cells through hypersensitivity to autologous or foreign antigens. It reduces and collagen and induces proliferation and inflammation of synovial fluid, resulting in cartilage and bone destruction.

Osteoarthritis refers to the degenerative condition of the synovial joint and is also called degenerative arthritis. It is a symptom of aging that occurs in middle or old age and causes movement disorders after 65 years of age. The main causes are thought to be caused by obesity, hypermotor function syndrome, recurrent dislocation, recurrent angioplasty, internal joint dislocation, inflammatory arthritis, and gout.

Osteoarthritis is mainly involved in the gradual damage to articular cartilage.When arthritis progresses, tumor necrosis factor-α (TNF-α), interleukin-1 β (IL-1 β), and nitric oxide are inflammatory cytokines. As the production of (NO) increases, it causes severe pain in tissues such as muscles, tendons, and ligaments.

Currently used arthritis treatment methods are largely divided into non-pharmaceutical treatment, drug treatment, and surgical treatment. Non-pharmaceutical treatment methods include reduction of joint load, patella fixation, heat treatment, and exercise. Drug treatment includes oral drugs and intra-articular injection treatment. The drugs used in the initial treatment are non-steroidal anti-inflammatory drugs (NSAIDs), and these NSAIDs simply reduce pain and relieve symptoms, but do not prevent joint cartilage loss or disease progression. None, and long-term use may cause heartburn and gastric bleeding as a side effect of gastrointestinal tract, kidney, heart, liver, and blood coagulation. Therefore, there is a need for research on natural products with fewer side effects.

Inflammatory bowel disease (IBD) is an incurable disease in which chronic inflammation or ulcers are caused in the mucous membranes of the large intestine and small intestine, and diarrhea and bloody stools continue for a long time, and recurrence is repeated.

Inflammatory bowel disease is a more common disease in Westerners, but it has been increasing rapidly in Korea since the 1980s, and the prevalent age of inflammatory bowel disease is 15-35 years old, and it is reported in all age groups, of which 15% are over 60 years old, and inflammatory. About 15% of patients with bowel disease have a family history in their immediate family.

The exact etiology of inflammatory bowel disease is still unclear, but it is estimated that the activation of inflammatory mediators and immune cells is an important etiology due to environmental or genetic factors as well as autoimmune diseases.

Inflammatory bowel disease is classified into two diseases, ulcerative colitis and Crohh's disease, which are clinically similar and differ from each other in terms of histological findings, endoscopic and immunological aspects, and these inflammatory bowel diseases are inflammatory mediated. It is known that the activation of factors and immune cells is an important etiology.

Persistent or inappropriate activation of the intestinal immune system plays an important role in the pathophysiology of chronic mucositis inflammation, especially due to infiltration of neutrophils, macrophages, lymphocytes and mast cells, eventually leading to mucosal destruction and ulcers.

In the inflammatory response that is mainly involved in the onset of inflammatory bowel disease, inflammation such as TNF-α (Tumor necrosis factor-α), interleukin-6 (IL-6), and interleukin-8 (IL-8) Response-promoting cytokines play a major role.

In particular, TNF-α is high in the colon lumen and colon epithelial cells of patients with ulcerative colitis, and according to recent studies, it is known that TNF-α plays an important role in the etiology of ulcerative colitis. Infliximab, an anti-TNF-α antibody, is known to be effective not only in the treatment of boils, but also in the treatment of Crohn's disease, which has not been treated previously. However, these treatments are expensive and cause side effects such as fluid reactions or infectious complications in some patients.

Currently, treatments for inflammatory bowel disease include 5-aminosalicylic acid (5-ASA) drugs that block the production of prostaglandins, such as sulfasalazine and mesalazine. In the case of using steroids or immunosuppressants such as steroids and not responding to steroid therapy, immunosuppressants such as azathioprine, 6-mercaptopurine, and Cyclosporin are also used, but there are no drugs that can expect a cure.

Alzheimer's disease is the most common cause of dementia and accounts for about 60 to 80% of dementia. About 33.9 million people worldwide have Alzheimer's, and the prevalence is expected to triple with an increase in life expectancy after 40 years. Currently, there is no medicine that can expect a clear improvement for Alzheimer's disease, and since it goes through a process of deteriorating for several decades after the onset, there is a trend of worldwide research on the prevention of Alzheimer's disease.

Alzheimer's disease is known to be caused by complex interactions of various risk factors. Along with genetic risk factors, socio-demographic risk factors such as age, sex and education, and environmental risk factors such as smoking, alcohol consumption, nutrition and social activity are also known. Representative genetic markers for Alzheimer's include APOE e4, but these genetic markers are not altered or modified. However, acquired factors (e.g., vascular risk factors, lifestyle habits) are modifiable factors, so studies investigating how these modifiable factors regulate the expression of dementia, along with reversing these factors, lowers the incidence of dementia or dementia. The timing of manifestation can be delayed.

Among Alzheimer's treatment drugs, Memantine was originally developed by Merz Pharmaceuticals in Germany in the 1980s as a treatment for Parkinson's disease and movement disorders, and has been marketed as a treatment for dementia in Germany. In 1999, as a result of a study that was effective in treating severe Alzheimer's disease was announced, it was approved as a treatment for Alzheimer's disease in Europe in 2002, and in October 2003, it was approved by the US FDA as a treatment for moderate to severe Alzheimer's disease. Recently, it is marketed as EBIXA® by Lundbeck in Korea. Commercially available Ebicsa contains memantine hydrochloride in the form of tablets or liquids, and the administration is carried out within a range not exceeding the maximum required dose of 20 mg per day in consideration of both aspects of efficacy and tolerance.

Memantine's mechanism of action is to block glutamate, an excitatory neurotransmitter, and glutamate excessively stimulates the nerves of the brain, causing excessive calcium inflow into the cranial nerve cells, causing damage and death of the cranial nerves. Memantine is known to prevent this. In Alzheimer's patients, it is known that glutamate is excessively secreted, so that the calcium ion channel of the NMDA receptor is always opened even in the stable period, and excessive calcium is introduced into the cells, which is associated with the death of the cranial nerve. Memantine acts as a low-affinity antagonist in the calcium transport channel located in the center of the NMDA receptor, blocking the inflow of calcium into the cells and preventing the destruction of cranial nerve cells. Excitement is reduced, and when physiological actions such as memory or learning are needed, it exits the calcium channel and depolarization occurs normally, stabilizing physiological nerve signals to normalize the function of nerve cells.

However, as side effects caused by memantine, loss of appetite, diarrhea, weight loss, dry mouth, dizziness, sleep disturbance, and fatigue have been reported. In addition, it has been reported to cause dizziness, headache, constipation, drowsiness, and high blood pressure in rare cases. Therefore, there is a need for research on natural products with fewer side effects.

Periodontitis (Periodontitis) is a type of inflammation that occurs in the supporting tissues around the teeth by toxins, which are metabolites of microorganisms living in the oral cavity. These periodontitis start from initial gingivitis, and if gingivitis is left untreated, the gums become swollen and develop into periodontitis accompanied by bleeding and severe bad breath. In addition, in the case of continuous periodontitis, the collagen supporting the fascia is destroyed and the alveolar bone supporting the tooth is dissolved, and the periodontal ligaments are separated to form a periodontal sac, and in severe cases, it develops into a progressive periodontal disease that damages the tooth.

The etiology of periodontitis differs according to sex, race, and age. This periodontal disease worsens for several years and repeats itself in a lull state, and the infection state persists, and the incidence rate is high in patients with systemic diseases such as diabetes, AIDS, neutropenia, and Down's syndrome.

The most effective method for preventing periodontitis can be achieved by using substances that inhibit the activity of these microorganisms and smooth blood flow to the inflamed gums. However, among the drugs used for the purpose of preventing and/or treating periodontitis, antibiotics such as penicillin, erythromycin, and tetracycline are effective for sterilization and growth inhibition, but have large side effects such as causing the appearance of resistant bacteria, such as chlorhexidine. The antibacterial agent has the effect of inhibiting the formation of cavities, but has a problem with strong toxicity. Therefore, there is a need for research on natural products with fewer side effects.

Endometriosis is a disease in which the endometrium or tissue similar thereto occurs outside the uterus depending on an increase in estrogen. Endometriosis is a benign disease, and causes pain including menstrual pain and lowering of dyspareunia, and significantly lowers the quality of life of women in social and reproductive activities. In endometriosis, menstrual pain appears to be very frequent, and pain symptoms such as lower abdominal pain, low back pain, dyspareunia, and bowel pain other than menstruation are also confirmed at high frequency.

There are many patients who repeat or recurrence until menopause unless radical surgery is performed for endometriosis, and treatment and management over a long period of time are required. In the treatment of endometriosis, drug therapy is often the first choice. Drug therapy is largely divided into symptomatic therapy and endocrine therapy. As a coping therapy, pain relievers and the like are mainly used for the purpose of improving pain caused by endometriosis. As an endocrine therapy, low-dose estrogen-progestin preparations, dienogest, gonadotropin-releasing hormone (GnRH) agonists, etc. are used for the purpose of suppressing estrogen-dependent endometrial proliferation in addition to improving pain.

However, in 10% to 30% of patients with endometriosis, it is said that pain medication cannot control pain caused by endometriosis. In addition, when using low-dose estrogen-progestin preparations, attention should be paid to thrombosis and liver failure. Dienogest is reported to have 71.9% of adverse events of negative stage bleeding in a long-term administration test, and may lead to severe anemia. The GnRH agonist is, in principle, not allowed for administration for more than 6 months, because there are cases where a decrease in the amount of bone salt based on an estrogen-lowering action is observed.

As described above, in the drug therapy for the treatment of endometriosis, there are many patients who have difficulty in continuing administration because the side effects peculiar to each drug are confirmed. Therefore, it is desired to develop new drugs that can reduce side effects and can be administered over a long period of time.

KR 10-1055920

KR 10-1292931

It is an object of the present invention to provide a melissa leaf extract fraction containing caffeic acid, EDPA (Ethyl 2-(3,4-dihydroxyphenyl) acetate), RME (Rosmarinic acid methyl ester), and rosmarinic acid.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of obesity comprising the extract fraction of melissa leaf as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of age-related macular degeneration, comprising the extract fraction of Melissa leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis comprising the extract fraction of melissa leaf as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of psoriasis comprising the extract fraction of Melissa leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer comprising the extract fraction of melissa leaf as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of diabetic retinopathy comprising the extract fraction of Melissa leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of glaucoma comprising the extract fraction of Melissa leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of Sjogren's syndrome comprising the extract fraction of melissa leaf as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating arteriosclerosis comprising a fraction of the extract of melissa leaf as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of arthritis comprising the extract fraction of Melissa leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory bowel disease comprising the extract fraction of Melissa leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of Alzheimer's, comprising the extract fraction of Melissa leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prophylaxis or treatment of periodontitis, comprising the extract fraction of Melissa leaf extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver comprising the extract fraction of melissa leaf as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of endometriosis, comprising the extract fraction of Melissa leaf extract as an active ingredient.

The present inventors are obesity, age-related macular degeneration, non-alcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren syndrome, arteriosclerosis, arthritis, inflammatory bowel disease, Alzheimer's, periodontitis, non-alcoholic fatty liver, prevention of endometriosis, or Recognizing the necessity of research for treatment, we attempted to achieve the purpose of research on natural substances that can prevent or treat these diseases.

Specifically, the inventors of the present invention are obesity, age-related macular degeneration, nonalcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren's syndrome, arteriosclerosis, arthritis , Inflammatory bowel disease, Alzheimer's, periodontitis, non-alcoholic fatty liver, found that there is an effect on the prevention and treatment of endometriosis, and completed the present invention.

Hereinafter, the present invention will be described in detail.

Melissa leaf extract fraction

The present invention is to provide a melissa leaf extract fraction containing caffeic acid, EDPA, RME and rosmarinic acid.

In an embodiment of the present invention, the Melissa leaf extract fraction is 0.1 to 5% by weight of caffeic acid, 0.1 to 5% by weight of EDPA, 0.01 to 2% by weight of RME, and 5 to 50% by weight of rosmarinic acid based on the total of the Melissa leaf extract fraction. It may contain, specifically (1) 0.2 to 4.0% by weight of caffeic acid, 0.15 to 4.0% by weight of EDPA, 0.02 to 1.5% by weight of RME, and 6 to 40% by weight of rosmarinic acid, and (2) caffeic acid 0.3 to 3.0 wt%, EDPA 0.2 to 3.5 wt%, RME 0.03 to 1.0 wt%, and rosmarinic acid 7 to 35 wt%, (3) caffeic acid 0.4 to 2.0 wt%, EDPA 0.3 to 3.0 wt% , 0.04 to 0.7% by weight of RME and 8 to 30% by weight of rosmarinic acid may be included, but the present invention is not limited thereto. Hereinafter, in the present specification, the fraction of the extract of Melissa leaf of the present invention may contain caffeic acid, EDPA, RME, and rosmarinic acid in the above-described weight% unless otherwise specified.

Melissa leaf extract fraction of the present invention is obesity, age-related macular degeneration, non-alcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren's syndrome, arteriosclerosis, arthritis, inflammatory bowel disease, Alzheimer's, periodontitis, non-alcoholic fatty liver, It is effective in preventing and treating endometriosis, and has excellent pharmacological effects while having low side effects.

Melissa leaf extract fraction of the present invention is obesity, age-related macular degeneration, non-alcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren's syndrome, arteriosclerosis, arthritis, inflammatory bowel disease, Alzheimer's, periodontitis, non-alcoholic fatty liver, It is effective in preventing and treating endometriosis, and has excellent pharmacological effects while having low side effects.

The fraction of the extract of Melissa leaf of the present invention does not contain rutin.

In an embodiment of the present invention, after reflux extraction of Melissa leaves with alcohol, the alcohol extract is concentrated, the concentrate is suspended with water, and fractions obtained by fractionating with ethyl acetate may be dried.

In an embodiment of the present invention, the fraction may be dried using a hot air drying method, a freeze drying method, or a spray drying method. For example, the fraction may be hot air dried using a hot air dryer, the fraction may be frozen and then freeze-dried by lowering the atmospheric pressure, or the fraction may be sprayed and spray dried by blowing hot air.

In the preparation of the extract fraction of melissa leaf of the present invention, dried melissa leaf or undried melissa leaf or a mixture thereof may be used. For effective extraction, the melissa leaves can be crushed and used.

In an embodiment of the present invention, the alcohol may be 70 to 80% (v/v) alcohol.

In the present invention, the term "alcohol" refers to a compound in which a hydroxyl group is bonded to a carbon atom of an alkyl or substituted alkyl group, and the term "alkyl" is a linear saturated hydrocarbon group or branched It means a saturated hydrocarbon group, and the term "substituted alkyl group" means that a substituent bonded to carbon of the alkyl group is substituted with hydroxy, cyano, halo, or the like.

The alcohol refers to a C1-C6 alcohol including 70 to 80% (v/v) ethanol, methanol, and the like, and may be preferably ethanol or methanol.

In the present invention, the term "C1-C6" refers to a functional group or main chain having 1 to 6 carbon atoms.

In an embodiment of the present invention, the alcohol extract may be obtained by the following steps, but is not limited thereto.

(S1) a first extraction step of obtaining a first extract by reflux extraction of Melissa leaves with 50-100% (v/v) alcohol at 80 to 85° C. for 2 to 6 hours;

(S2) a second extraction step of obtaining a second extract by reflux extraction of the remaining Melissa residue after the first extraction at 80 to 85° C. with 50 to 100% (v/v) alcohol for 2 to 6 hours; And

(S3) mixing the first extract and the second extract.

The alcohol extract can be obtained by mixing the first extract and the second extract through the first and second extraction steps above.

To prepare the alcohol extract, using 50 to 100% alcohol (v/v), preferably 70 to 80% alcohol (v/v) of 5 to 15 times (v/w) of Melissa leaf Can be extracted.

In an embodiment of the present invention, the concentrate may be obtained by the following steps, but is not limited thereto.

(S4) a first concentration step of obtaining a first concentrate by concentrating the alcohol extract for 5 to 15 hours under a temperature condition of 50 to 60° C. and a pressure condition of -0.066 to -0.070 MPa, and

(S5) A second concentration step of obtaining a second concentrate by concentrating the first concentrate for 3 to 10 hours under a temperature condition of 55 to 60° C. and a pressure condition of -0.063 to -0.065 MPa.

In an embodiment of the present invention, before drying the fraction, the following concentration step may be further included, but is not limited thereto.

(S6) the third concentration step of concentrating the fraction.

The third concentration step of concentrating the fractions for 3 to 10 hours under a temperature condition of 55 to 60° C. and a pressure condition of -0.063 to -0.065 MPa.

In the present invention, the term "fraction" means a result obtained by performing fractionation in order to separate a specific component or a specific component group from a mixture containing various various constituents.

In the present invention, the fractionation method for obtaining the fraction of the extract is not particularly limited, and may be performed according to a method commonly used in the art. Specifically, a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing an ultrafiltration membrane having a certain molecular weight cut-off value, and various chromatography (separation according to size, charge, hydrophobicity or affinity) And a chromatographic fractionation method for performing a method, and combinations thereof. Specifically, there may be mentioned a method of obtaining a fraction from the extract by treating the extract obtained by extracting the Melissa leaf of the present invention with a predetermined solvent.

In the present invention, the type of fractionation solvent used to obtain the fraction is not particularly limited, and any solvent known in the art may be used, and specifically, ethyl acetate, which is a non-polar solvent.

The fraction of the melissa leaf extract may be an ethyl acetate fraction of the ethanol extract of melissa leaf.

The above method effectively extracts water-soluble substances and non-aqueous substances by using 50 to 100% alcohol (v/v), and the solubility in water is low, but dissolution in ethyl acetate enables extraction of high water-insoluble substances. There is.

The ethyl acetate fraction of the ethanol extract of Melissa leaf may include 0.1 to 5% by weight of caffeic acid, 0.1 to 5% by weight of EDPA, 0.01 to 2% by weight of RME, and 5 to 50% by weight of rosmarinic acid based on the total weight of the fraction. More specifically, (1) 0.2 to 4.0% by weight of caffeic acid, 0.15 to 4.0% by weight of EDPA, 0.02 to 1.5% by weight of RME, and 6 to 40% by weight of rosmarinic acid, and (2) 0.3 to 3.0% by weight of caffeic acid , EDPA 0.2 to 3.5% by weight, RME 0.03 to 1.0% by weight, and may contain 7 to 35% by weight of rosmarinic acid, (3) caffeic acid 0.4 to 2.0% by weight, EDPA 0.3 to 3.0% by weight, RME 0.04 to 0.7% by weight % And 8 to 30% by weight of rosmarinic acid.

Advantages and features of the present invention, and a method of achieving them will become apparent with reference to the embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in a variety of different forms. It is provided to completely inform the scope of the invention to the possessor, and the invention is only defined by the scope of the claims.

Pharmaceutical composition for the prevention or treatment of obesity comprising the extract fraction of Melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of obesity comprising the extract fraction of melissa leaf as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "obesity" refers to a state in which fat is excessively accumulated in the body, resulting in an increase in weight beyond the limits of skeletal and physical demands.

In the present invention, the term ``active ingredient'' includes a substance or group of substances that is expected to directly or indirectly express the efficacy effect of the composition by its inherent pharmacological action (including herbal medicines for which pharmacologically active ingredients, etc. are not known. It means to include the main component as ).

The pharmaceutical composition for the prevention or treatment of obesity comprising the melissa leaf extract fraction of the present invention as an active ingredient contains the melissa leaf extract fraction as an active ingredient, and may further include a pharmaceutically acceptable carrier. Depending on the method, it may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, oral dosage forms such as aerosols, external preparations, and sterile injectable solutions.

In the present invention, the term "pharmaceutically acceptable" refers to a composition that is physiologically acceptable and does not usually cause an allergic reaction such as gastrointestinal disorders or dizziness or similar reactions when administered to humans.

In the present invention, the term "pharmaceutically acceptable carrier" typically includes a liquid or non-liquid basis of a pharmaceutical composition. If the pharmaceutical composition is provided in liquid form, the carrier typically comprises pyrogen-free water; Isotonic saline or buffered (aqueous) solutions, such as phosphate, citric acid, etc. buffer solutions. The injection buffer can be on a hypertonic, isotonic or hypotonic basis for a specific reference media, i.e. the buffer can have a high, the same or a low salt content for a specific reference medium, preferably osmotic pressure or other concentration effects. Such concentrations of the aforementioned salts, which do not induce cell damage by, can be used. The reference medium is blood, lymph, cytoplasmic liquid, or other bodily fluid, or a body fluid that can be used as a reference medium in the "ex vivo" method, for example as a general buffer or liquid, arising from, for example, "in vivo" methods. It's a liquid. Such general buffers or liquids are known to the person skilled in the art.

The pharmaceutically acceptable carriers are those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.

In addition, the pharmaceutical composition of the present invention may contain diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and other pharmaceutically acceptable additives.

The pharmaceutical composition of the present invention may be prepared in the form of a liquid, suspension, powder, granule, tablet, capsule, pill, or extract.

The composition of the present invention may be administered orally or parenterally (eg, application or intravenous, subcutaneous, intraperitoneal injection).

In the present invention, the term "oral administration" is a method of injecting a drug for improving pathological symptoms by mouth, and in the present invention, the term "parenteral administration" refers to subcutaneous, intramuscular, and It refers to a method of intraperitoneal administration using an intravenous or tube.

Solid preparations for oral administration include powders, granules, tablets, capsules, soft capsules, pills, and the like. Liquid preparations for oral administration include suspensions, liquid contents, emulsions, syrups, aerosols, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. May be included.

Formulations for parenteral administration include aqueous solutions, liquids, non-aqueous solutions, suspensions, emulsions, eye drops, ointments, syrups, suppositories, aerosols, and other external preparations and sterile injections, each sterilized according to a conventional method. It may be used in combination, and preferably, a pharmaceutical composition of cream, gel, patch, spray, ointment, warning agent, lotion, liniment, eye ointment, eye drop, pasta or cataplasma may be prepared and used. However, it is not limited thereto. Compositions for topical administration may be anhydrous or aqueous, depending on the clinical prescription. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like can be used.

The pharmaceutically acceptable additive according to the present invention may include 0.1 to 99.9 parts by weight, specifically 0.1 to 50 parts by weight, with respect to the composition, but is not limited thereto.

The pharmaceutical composition for preventing or treating obesity containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

In the case of a pharmaceutical composition for the prevention or treatment of obesity comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it exhibits remarkable weight suppression, blood triglyceride suppression, blood sugar suppression, and total cholesterol suppression effect to prevent or treat obesity. There is.

The pharmaceutical composition for preventing or treating obesity containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal It can be administered in a conventional manner through the route, and specifically, it can be administered orally.

The pharmaceutical composition for preventing or treating obesity containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered in a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. .

A pharmaceutical composition for the prevention or treatment of age-related macular degeneration comprising a fraction of melissa leaf extract as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of age-related macular degeneration, comprising the extract fraction of Melissa leaf extract as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term ``macular degeneration'' refers to a disease in which the macular deteriorates due to aging, genetic factors, toxicity, inflammation, etc., resulting in decreased visual acuity, and in severe cases, complete loss of vision. .

In the case of a pharmaceutical composition for preventing or treating age-related macular degeneration comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it exhibits an effect of suppressing the size of macular degeneration lesions, thereby preventing or treating age-related macular degeneration. .

The pharmaceutical composition for the prevention or treatment of age-related macular degeneration containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for the prevention or treatment of age-related macular degeneration containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular Alternatively, it may be administered in a conventional manner through the intradermal route, and specifically, it may be administered orally.

The pharmaceutical composition for the prevention or treatment of age-related macular degeneration containing the extract fraction of Melissa leaf of the present invention as an active ingredient is administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. desirable.

As long as the matters mentioned in the pharmaceutical composition for preventing or treating obesity containing the Melissa leaf extract fraction as an active ingredient are not contradictory to each other, the pharmaceutical for the prevention or treatment of age-related macular degeneration containing the Melissa leaf extract fraction as an active ingredient The same applies to the composition.

Pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis comprising the extract fraction of Melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis comprising the extract fraction of melissa leaf as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "non-alcoholic steatohepatitis" refers to a disease having a form similar to that of alcoholic liver disorder even though there is no drinking history.

In the case of a pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, remarkable fibrosis inhibitory effect, Col1A2, TGF beta, inhibition of protein expression levels of IL-6 and IL-10 It has an effect that can prevent or treat non-alcoholic steatohepatitis.

The pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, ocular It may be administered in a conventional manner through an intraoral or intradermal route, and specifically, it may be administered orally.

The pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. It is desirable.

As long as the matters mentioned in the pharmaceutical composition for preventing or treating obesity containing the melissa leaf extract fraction as an active ingredient do not contradict each other, the pharmaceutical for the prevention or treatment of non-alcoholic steatohepatitis containing the melissa leaf extract fraction as an active ingredient The same applies to the appropriate composition.

Pharmaceutical composition for the prevention or treatment of psoriasis comprising a fraction of melissa leaf extract as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of psoriasis comprising the extract fraction of Melissa leaf extract as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "psoriasis" refers to a chronic inflammatory skin disease characterized by clear bordered papules and plaques covered with silvery white scales.

In the case of a pharmaceutical composition for the prevention or treatment of psoriasis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it is possible to prevent or treat psoriasis by suppressing remarkable epidermal thickness, inflammatory cell infiltration of ear tissue, and mRNA expression level of ear tissue. There is an effect.

The pharmaceutical composition for preventing or treating psoriasis containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for the prevention or treatment of psoriasis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal It can be administered in a conventional manner through the route, and specifically, it can be administered orally.

The pharmaceutical composition for the prevention or treatment of psoriasis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered in a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. .

Matters mentioned in the pharmaceutical composition for the prevention or treatment of obesity containing the melissa leaf extract fraction as an active ingredient are not contradictory to each other, even in the pharmaceutical composition for the prevention or treatment of psoriasis containing the melissa leaf extract fraction as an active ingredient The same applies.

Pharmaceutical composition for the prevention or treatment of cancer comprising the extract fraction of melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer comprising the extract fraction of melissa leaf as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "cancer" refers to a disease in which the cell cycle is not regulated and thus cell division continues.

In the case of a pharmaceutical composition for preventing or treating cancer comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the size of the cancer tumor is significantly reduced, thereby preventing or treating cancer.

The pharmaceutical composition for preventing or treating cancer containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The cancer is lung cancer, non-small cell lung cancer (NSCL), bronchial alveolar cell lung cancer, gastric cancer, gastrointestinal cancer, liver cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or eye melanoma, ovarian cancer, rectal cancer, colon cancer, colon cancer, Breast cancer, fallopian duct carcinoma, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, esophageal cancer, laryngeal cancer, small intestine cancer, thyroid cancer, parasensitivity carcinoma, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, multiple myeloma, chronic or acute leukemia, childhood It may be any one of solid tumor, lymphoma, bladder cancer, kidney cancer, renal cell carcinoma, renal pelvic carcinoma, contractile tumor, brain stem glioma, or pituitary adenoma, and specifically colon cancer.

The pharmaceutical composition for the prevention or treatment of cancer containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal It can be administered in a conventional manner through the route, and specifically, it can be administered orally.

The pharmaceutical composition for the prevention or treatment of cancer containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. .

Matters mentioned in the pharmaceutical composition for the prevention or treatment of obesity containing the extract fraction of Melissa leaf as an active ingredient are not contradictory to each other, even in the pharmaceutical composition for the prevention or treatment of cancer containing the extract fraction of Melissa leaf as an active ingredient The same applies.

Pharmaceutical composition for preventing or treating diabetic retinopathy comprising a fraction of melissa leaf extract as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of diabetic retinopathy comprising the extract fraction of Melissa leaf extract as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "diabetic retinopathy" refers to a state in which blood vessels in the retina in our eyes are damaged by diabetes.

In the case of a pharmaceutical composition for preventing or treating diabetic retinopathy comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it has an effect of preventing or treating diabetic retinopathy because it has an excellent inhibitory effect on retinal vessel leakage.

The pharmaceutical composition for preventing or treating diabetic retinopathy containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for preventing or treating diabetic retinopathy containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular Alternatively, it may be administered in a conventional manner through the intradermal route, and specifically, it may be administered orally.

The pharmaceutical composition for preventing or treating diabetic retinopathy containing the extract fraction of Melissa leaf of the present invention as an active ingredient is administered in a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. desirable.

As long as the matters mentioned in the pharmaceutical composition for preventing or treating obesity containing the melissa leaf extract fraction as an active ingredient are not contradictory to each other, the pharmaceutical for the prevention or treatment of diabetic retinopathy containing the melissa leaf extract fraction as an active ingredient The same applies to the composition.

Pharmaceutical composition for preventing or treating glaucoma comprising a fraction of melissa leaf extract as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of glaucoma comprising the extract fraction of Melissa leaf extract as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "glaucoma" refers to a disease occurring in the optic nerve including retinal plexus cells while taking the form of optic nerve atrophy.

In the case of a pharmaceutical composition for preventing or treating glaucoma comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, an increase in intraocular pressure is significantly suppressed, thereby preventing or treating glaucoma.

The pharmaceutical composition for preventing or treating glaucoma containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for preventing or treating glaucoma containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal It can be administered in a conventional manner through the route, and specifically, it can be administered orally.

The pharmaceutical composition for preventing or treating glaucoma containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. .

Matters mentioned in the pharmaceutical composition for the prevention or treatment of obesity containing the melissa leaf extract fraction as an active ingredient are not contradictory to each other, even in the pharmaceutical composition for the prevention or treatment of glaucoma containing the melissa leaf extract fraction as an active ingredient The same applies.

Pharmaceutical composition for the prevention or treatment of Sjogren's syndrome comprising the extract fraction of Melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of Sjogren's syndrome comprising the extract fraction of melissa leaf as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "Sjogren's syndrome" refers to an autoimmune disease in which the human body's immune system attacks glands (glands) that produce moisture (water).

In the case of a pharmaceutical composition for preventing or treating Sjogren's syndrome comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the amount of saliva secretion and the amount of tear secretion increase, thereby preventing or treating Sjogren's syndrome.

The pharmaceutical composition for the prevention or treatment of Sjogren's syndrome containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for the prevention or treatment of Sjogren's syndrome containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or It can be administered in a conventional manner through the intradermal route, and specifically, it can be administered orally.

The pharmaceutical composition for the prevention or treatment of Sjogren's syndrome containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

As long as the matters mentioned in the pharmaceutical composition for preventing or treating obesity containing the melissa leaf extract fraction as an active ingredient do not contradict each other, a pharmaceutical composition for the prevention or treatment of Sjogren's syndrome comprising the melissa leaf extract fraction as an active ingredient The same applies to

Pharmaceutical composition for preventing or treating arteriosclerosis comprising a fraction of melissa leaf extract as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating arteriosclerosis comprising a fraction of the extract of melissa leaf as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "arteriosclerosis" refers to a disease in which cholesterol or triglycerides accumulate in the innermost membrane of blood vessels, the blood vessels are narrowed and hardened and blocked.

In the case of a pharmaceutical composition for preventing or treating arteriosclerosis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the increase in atherosclerotic plaque is significantly suppressed, thereby preventing or treating arteriosclerosis.

The pharmaceutical composition for preventing or treating arteriosclerosis containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for preventing or treating arteriosclerosis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or It can be administered in a conventional manner through the intradermal route, and specifically, it can be administered orally.

The pharmaceutical composition for preventing or treating arteriosclerosis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

As long as the matters mentioned in the pharmaceutical composition for preventing or treating obesity containing the melissa leaf extract fraction as an active ingredient are not contradictory to each other, a pharmaceutical composition for preventing or treating arteriosclerosis comprising the melissa leaf extract fraction as an active ingredient The same applies to

Pharmaceutical composition for the prevention or treatment of arthritis comprising the extract fraction of Melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of arthritis comprising the extract fraction of Melissa leaf extract as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "arthritis" refers to a disease in which damage or inflammation occurs due to various causes in a joint, which is a site where bone and bone meet.

In the case of a pharmaceutical composition for preventing or treating arthritis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it has an excellent arthritis treatment effect and an excellent weight-bearing increase effect, thereby preventing or treating arthritis.

The pharmaceutical composition for preventing or treating arthritis containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The arthritis may be any one of osteoarthritis, degenerative arthritis, detachable osteochondritis, joint ligament injury, psoriatic arthritis, ankylosing spondylitis and rheumatoid arthritis, and specifically osteoarthritis and rheumatoid arthritis.

The pharmaceutical composition for preventing or treating arthritis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal It can be administered in a conventional manner through the route, and specifically, it can be administered orally.

The pharmaceutical composition for the prevention or treatment of arthritis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. .

Matters mentioned in the pharmaceutical composition for the prevention or treatment of obesity containing the extract fraction of Melissa leaf as an active ingredient are not contradictory to each other, even in the pharmaceutical composition for the prevention or treatment of arthritis containing the extract fraction of Melissa leaf as an active ingredient The same applies.

Pharmaceutical composition for the prevention or treatment of inflammatory bowel disease comprising the extract fraction of melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory bowel disease comprising the extract fraction of Melissa leaf extract as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "inflammatory bowel disease" refers to a disease in which abnormal chronic inflammation in the intestine repeats improvement and recurrence.

In the case of a pharmaceutical composition for preventing or treating inflammatory bowel disease comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it has an effect of preventing or treating inflammatory bowel disease because it exhibits a low disease activity index.

The pharmaceutical composition for preventing or treating inflammatory bowel disease containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The inflammatory bowel disease may be any one of ulcerative colitis, Crohn's disease, intestinal tuberculosis, ischemic colitis, and intestinal ulcers due to Behcet's disease, and specifically, may be ulcerative colitis.

The pharmaceutical composition for the prevention or treatment of inflammatory bowel disease containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular Alternatively, it may be administered in a conventional manner through the intradermal route, and specifically, it may be administered orally.

The pharmaceutical composition for preventing or treating inflammatory bowel disease containing the extract fraction of Melissa leaf of the present invention as an active ingredient is administered in a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. desirable.

As long as the matters mentioned in the pharmaceutical composition for preventing or treating obesity containing the melissa leaf extract fraction as an active ingredient are not contradictory to each other, the pharmaceutical for the prevention or treatment of inflammatory bowel disease containing the melissa leaf extract fraction as an active ingredient The same applies to the composition.

Pharmaceutical composition for the prevention or treatment of Alzheimer's containing the extract fraction of Melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of Alzheimer's, comprising the extract fraction of Melissa leaf extract as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "alzheimer" refers to the most common cause of dementia, which causes dementia symptoms due to degenerative changes in brain cells.

In the case of a pharmaceutical composition for the prevention or treatment of Alzheimer, comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, cognitive function is improved, thereby preventing or treating Alzheimer's.

The pharmaceutical composition for the prevention or treatment of Alzheimer's containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for the prevention or treatment of Alzheimer's containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal It can be administered in a conventional manner through the route, and specifically, it can be administered orally.

The pharmaceutical composition for the prophylaxis or treatment of Alzheimer, containing the extract fraction of Melissa leaf of the present invention as an active ingredient, is preferably administered in a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. .

Matters mentioned in the pharmaceutical composition for the prevention or treatment of obesity containing the melissa leaf extract fraction as an active ingredient are not contradictory to each other, even in the pharmaceutical composition for the prevention or treatment of Alzheimer's containing the melissa leaf extract fraction as an active ingredient The same applies.

Pharmaceutical composition for the prevention or treatment of periodontitis comprising the extract fraction of Melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prophylaxis or treatment of periodontitis, comprising the extract fraction of Melissa leaf extract as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "periodontal" refers to diseases appearing in tissues around teeth, such as the gingiva, periodontal ligaments and alveolar bones surrounding the teeth.

In the case of a pharmaceutical composition for the prevention or treatment of periodontitis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the expression of pro-collagen increases, thereby effectively regenerating the gum tissue, thereby having a significant therapeutic effect on periodontitis. There is.

The pharmaceutical composition for preventing or treating periodontitis containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for the prevention or treatment of periodontitis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal It can be administered in a conventional manner through the route, and specifically, it can be administered orally.

The pharmaceutical composition for the prevention or treatment of periodontitis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered in a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. .

Matters mentioned in the pharmaceutical composition for the prevention or treatment of obesity containing the extract fraction of Melissa leaf as an active ingredient are not contradictory to each other, even in the pharmaceutical composition for the prevention or treatment of periodontitis containing the extract fraction of Melissa leaf as an active ingredient The same applies.

Pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver comprising the extract fraction of Melissa leaf as an active ingredient

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver comprising the extract fraction of melissa leaf as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

In the present invention, the term "non-alcoholic fatty liver disease" refers to a disease that exhibits a form similar to that of alcoholic fatty liver even though there is no alcoholic fatty liver disease.

In the case of a pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it exhibits remarkable weight suppression, blood triglyceride suppression, blood sugar suppression, and total cholesterol suppression effect to prevent or It has a curative effect.

The pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver containing the melissa leaf extract fraction as an active ingredient has a melissa leaf extract fraction in an amount of 20% to 80% by weight based on the total weight.

The pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular Alternatively, it may be administered in a conventional manner through the intradermal route, and specifically, it may be administered orally.

The pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver containing the extract fraction of Melissa leaf of the present invention as an active ingredient is administered at a dose of 0.001 mg/kg to 50 mg/kg once to several times a day. desirable.

As long as the matters mentioned in the pharmaceutical composition for preventing or treating obesity containing the melissa leaf extract fraction as an active ingredient are not contradictory to each other, the pharmaceutical for the prevention or treatment of non-alcoholic fatty liver containing the melissa leaf extract fraction as an active ingredient The same applies to the composition.

Pharmaceutical composition for the prevention or treatment of endometriosis containing the extract fraction of Melissa leaf as an active ingredient

The present invention is to provide a pharmaceutical composition for the prevention or treatment of endometriosis comprising the extract fraction of Melissa leaf as an active ingredient.

In the present invention, the term "endometriosis" refers to a disease in which various symptoms such as pain and bleeding occur due to the presence of endometrial tissue in the ovary, posterior wall of the uterus, uterine ligaments, and pelvic wall.

In the case of a pharmaceutical composition for preventing or treating endometriosis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the size of the uterine lesion is significantly reduced, thereby preventing or treating endometriosis.

The pharmaceutical composition for preventing or treating endometriosis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or It can be administered in a conventional manner through the intradermal route, and specifically, it can be administered orally. The pharmaceutical composition for preventing or treating endometriosis containing the extract fraction of Melissa leaf of the present invention as an active ingredient is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. The Melissa leaf extract fraction is the same as described above.

Food composition containing the extract fraction of melissa leaf as an active ingredient

Another object of the present invention is to provide a food composition comprising a melissa leaf extract fraction as an active ingredient.

The fraction of the Melissa leaf extract and its preparation method are the same as those of salpin.

The composition is any one of obesity, age-related macular degeneration, nonalcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren syndrome, arteriosclerosis, arthritis, inflammatory bowel disease, Alzheimer's periodontitis, nonalcoholic fatty liver, and endometriosis. It is to prevent or improve.

The cancer is lung cancer, non-small cell lung cancer (NSCL), bronchial alveolar cell lung cancer, gastric cancer, gastrointestinal cancer, liver cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or eye melanoma, ovarian cancer, rectal cancer, colon cancer, colon cancer, Breast cancer, fallopian duct carcinoma, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, esophageal cancer, laryngeal cancer, small intestine cancer, thyroid cancer, parasensitivity carcinoma, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, multiple myeloma, chronic or acute leukemia, childhood It may be any one of solid tumor, lymphoma, bladder cancer, kidney cancer, renal cell carcinoma, renal pelvic carcinoma, contractile tumor, brain stem glioma, or pituitary adenoma, and specifically colon cancer.

The arthritis may be any one of osteoarthritis, degenerative arthritis, detachable osteochondritis, joint ligament injury, psoriatic arthritis, ankylosing spondylitis and rheumatoid arthritis, and specifically osteoarthritis and rheumatoid arthritis.

In the food composition according to the present invention, when the extract fraction of Melissa leaf is used as an additive to the food composition, it may be added as it is or may be used with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient can be appropriately determined according to each purpose of use, such as prevention, health or treatment.

The formulation of the food composition may be in the form of powders, granules, pills, tablets, capsules, as well as general foods or beverages.

The type of food is not particularly limited, and examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , Various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and may include all foods in the usual sense.

In general, in the manufacture of food or beverage, the extract fraction of Melissa leaf may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on 100 parts by weight of raw materials. However, in the case of long-term intake for the purpose of health and hygiene or for health control purposes, the amount may be less than or equal to the above range, and the present invention has no problem in terms of safety in terms of using fractions from natural products. It can also be used in the above amount.

In the food composition according to the present invention, the beverage may contain various flavoring agents or natural carbohydrates as an additional component, like a conventional beverage. The natural carbohydrates described above may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the beverage according to the present invention.

In addition to the above, the food composition according to the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain carbonates used in alcohol and carbonated beverages. In addition, the composition for improving sleep of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These ingredients may be used independently or in combination. The ratio of these additives is not limited, but it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the food composition of the present invention.

Prevention or treatment method

Another object of the present invention is to provide a method for preventing or treating obesity by administering a therapeutically effective amount of an extract fraction of melissa leaf to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating age-related macular degeneration by administering a therapeutically effective amount of an extract fraction of Melissa leaf to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating psoriasis by administering a therapeutically effective amount of the extract fraction of Melissa leaf extract to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating cancer by administering a therapeutically effective amount of the extract fraction of melissa leaf to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating diabetic retinopathy by administering a therapeutically effective amount of the extract fraction of melissa leaf to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating glaucoma by administering a therapeutically effective amount of an extract fraction of Melissa leaf to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating Sjogren's syndrome by administering a therapeutically effective amount of a fraction of a melissa leaf extract to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating arteriosclerosis by administering a therapeutically effective amount of the extract fraction of Melissa leaf to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating arthritis by administering a therapeutically effective amount of the extract fraction of melissa leaf to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating inflammatory bowel disease by administering a therapeutically effective amount of melissa leaf extract fraction to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating Alzheimer's by administering a therapeutically effective amount of the extract fraction of Melissa leaf extract to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating periodontitis by administering a therapeutically effective amount of the extract fraction of Melissa leaf extract to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating non-alcoholic fatty liver by administering a therapeutically effective amount of the extract fraction of melissa leaf to a subject in need of treatment.

Another object of the present invention is to provide a method for preventing or treating endometriosis by administering a therapeutically effective amount of an extract fraction of Melissa leaf to a subject in need of treatment.

In the present specification, the term "subject in need of treatment" refers to mammals including humans, and the term "administration" refers to providing a predetermined substance to a patient by any suitable method. In the treatment method of the present invention, the pharmaceutical composition containing the extract fraction of Melissa leaf of the present invention as an active ingredient is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal It can be administered in a conventional manner via the route.

In the method for preventing or treating obesity of the present invention, the extract fraction of Melissa leaf of the present invention is a conventional oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal route. It can be administered in a manner, and specifically, it can be administered orally.

In addition, in the method for preventing or treating age-related macular degeneration of the present invention, the extract fraction of Melissa leaf of the present invention can be used for oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a conventional manner, and specifically, it can be administered orally.

In addition, in the method of preventing or treating non-alcoholic steatohepatitis of the present invention, the extract fraction of Melissa leaf of the present invention is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal route It can be administered in a conventional manner through, specifically, it can be administered orally.

In addition, in the method for preventing or treating psoriasis of the present invention, the extract fraction of Melissa leaf of the present invention is conventionally used through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a phosphorus manner, and specifically, it can be administered orally.

In addition, in the method for preventing or treating cancer of the present invention, the extract fraction of Melissa leaf of the present invention is conventionally used through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a phosphorus manner, and specifically, it can be administered orally.

In addition, in the method for preventing or treating diabetic retinopathy of the present invention, the extract fraction of Melissa leaf of the present invention can be used for oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a conventional manner, and specifically, it can be administered orally.

In addition, in the method for preventing or treating glaucoma of the present invention, the extract fraction of Melissa leaf of the present invention is conventionally used through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a phosphorus manner, and specifically, it can be administered orally.

In addition, in the method for preventing or treating Sjogren's syndrome of the present invention, the extract fraction of Melissa leaf of the present invention is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal route. It can be administered in a conventional manner, and specifically, it can be administered orally.

In addition, in the method of preventing or treating arteriosclerosis of the present invention, the extract fraction of Melissa leaf of the present invention is oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal route. It can be administered in a conventional manner, and specifically, it can be administered orally.

In addition, in the method for preventing or treating arthritis of the present invention, the extract fraction of Melissa leaf of the present invention is conventionally used through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a phosphorus manner, and specifically, it can be administered orally.

In addition, in the method of preventing or treating inflammatory bowel disease of the present invention, the extract fraction of Melissa leaf of the present invention can be used for oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a conventional manner, and specifically, it can be administered orally.

In addition, in the method for preventing or treating Alzheimer of the present invention, the extract fraction of Melissa leaf of the present invention is conventionally used through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a phosphorus manner, and specifically, it can be administered orally.

In the method for preventing or treating periodontitis of the present invention, the extract fraction of Melissa leaf of the present invention is a conventional oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal route. It can be administered in a manner, and specifically, it can be administered orally.

In addition, in the method of preventing or treating non-alcoholic fatty liver of the present invention, the extract fraction of Melissa leaf of the present invention can be used for oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a conventional manner, and specifically, it can be administered orally.

In the method for preventing or treating endometriosis of the present invention, the extract fraction of Melissa leaf of the present invention is conventionally used through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a phosphorus manner, and specifically, it can be administered orally.

The term "therapeutically effective amount" as used herein refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal, or human thought by a researcher, veterinarian, doctor, or other clinician. , Which includes an amount that induces relief of symptoms of the disease or disorder being treated. It is apparent to those skilled in the art that the therapeutically effective dosage and frequency of administration of the active ingredient of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient. It can be adjusted according to various factors including condition, sex and diet, time of administration, route of administration and rate of secretion of the composition, duration of treatment, and drugs used concurrently.

In the method for preventing or treating obesity of the present invention, in the case of adults, the extract fraction of Melissa leaf of the present invention is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

In the method for preventing or treating age-related macular degeneration of the present invention, in the case of adults, the extract fraction of Melissa leaf of the present invention is administered at a dose of 0.001 mg/kg to 50 mg/kg once to several times a day. It is desirable.

In the method of preventing or treating nonalcoholic steatohepatitis of the present invention, in the case of adults, the Melissa leaf extract fraction of the present invention is administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. It is desirable to do it.

In the method for preventing or treating psoriasis of the present invention, in the case of an adult, the extract fraction of Melissa leaf of the present invention is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

In the method of preventing or treating cancer of the present invention, in the case of an adult, the extract fraction of Melissa leaf of the present invention is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

In the method for preventing or treating diabetic retinopathy of the present invention, in the case of adults, the extract fraction of Melissa leaf of the present invention is administered at a dose of 0.001 mg/kg to 50 mg/kg once to several times a day. It is desirable.

In the method of preventing or treating glaucoma of the present invention, in the case of an adult, the extract fraction of Melissa leaf of the present invention is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

In the method of preventing or treating Sjogren's syndrome of the present invention, in the case of adults, the extract fraction of Melissa leaf of the present invention is administered at a dose of 0.001 mg/kg to 50 mg/kg once to several times a day. desirable.

In the method for preventing or treating arteriosclerosis of the present invention, in the case of an adult, the extract fraction of Melissa leaf of the present invention is administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. desirable.

In the method of preventing or treating arthritis of the present invention, in the case of an adult, the extract fraction of Melissa leaf of the present invention is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

In the method of preventing or treating inflammatory bowel disease of the present invention, in the case of an adult, the extract fraction of Melissa leaf of the present invention is administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. It is desirable.

In the method of preventing or treating Alzheimer of the present invention, in the case of adults, the extract fraction of Melissa leaf of the present invention is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

In the method for preventing or treating periodontitis of the present invention, in the case of adults, the extract fraction of Melissa leaf of the present invention is preferably administered at a dose of 0.001 mg/kg to 50 mg/kg when administered once to several times a day. Do.

In the method for preventing or treating non-alcoholic fatty liver of the present invention, in the case of adults, the extract fraction of Melissa leaf of the present invention is administered at a dose of 0.001 mg/kg to 50 mg/kg once to several times a day. It is desirable.

In the method for preventing or treating endometriosis of the present invention, in the case of an adult, the extract fraction of Melissa leaf of the present invention is administered at a dose of 0.001 mg/kg to 50 mg/kg once to several times a day. desirable.

In the present invention, the term "prevention" means obesity, age-related macular degeneration, nonalcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren's syndrome, arteriosclerosis, Any action that inhibits or delays arthritis, inflammatory bowel disease, Alzheimer's periodontitis, non-alcoholic fatty liver, or endometriosis.

In the present invention, the term ``treatment'' refers to obesity, age-related macular degeneration, nonalcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren syndrome, arteriosclerosis, Arthritis, inflammatory bowel disease, Alzheimer's, periodontitis, non-alcoholic fatty liver, or endometriosis is any action that improves or beneficially alters.

Use of a pharmaceutical composition comprising a fraction of melissa leaf extract for prophylaxis or treatment as an active ingredient

It is an object of the present invention to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of obesity.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of age-related macular degeneration.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising an extract fraction of Melissa leaf for the prophylaxis or treatment of non-alcoholic steatohepatitis as an active ingredient.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of psoriasis.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of cancer.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising an extract fraction of Melissa leaf for the prevention or treatment of diabetic retinopathy as an active ingredient.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of glaucoma.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of Sjogren's syndrome.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of arteriosclerosis.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising an extract fraction of Melissa leaf for the prevention or treatment of arthritis as an active ingredient.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of inflammatory bowel disease.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of Alzheimer's.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of periodontitis.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of non-alcoholic fatty liver.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the prevention or treatment of endometriosis.

Use of a pharmaceutical composition comprising the extract fraction of melissa leaf for the manufacture of drugs as an active ingredient

An object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for preventing or treating obesity.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of age-related macular degeneration.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of non-alcoholic steatohepatitis.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of psoriasis.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of cancer.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for preventing or treating diabetic retinopathy.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for preventing or treating glaucoma.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of Sjogren's syndrome.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for preventing or treating arteriosclerosis.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of arthritis.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of inflammatory bowel disease.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of Alzheimer's.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for the prevention or treatment of periodontitis.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for preventing or treating non-alcoholic fatty liver.

Another object of the present invention is to provide a use of a pharmaceutical composition comprising a melissa leaf extract fraction as an active ingredient for the manufacture of a medicament for preventing or treating endometriosis.

In one embodiment of the present invention, the pharmaceutical composition comprising the extract fraction of Melissa leaf for the manufacture of a drug as an active ingredient may be mixed with an acceptable carrier, etc., and may further include other agents.

Matters mentioned in the pharmaceutical composition, treatment method, and use of the present invention are the same as long as they do not contradict each other.

The melissa leaf extract fraction of the present invention has superior obesity, age-related macular degeneration, nonalcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren's syndrome, arteriosclerosis, arthritis, inflammatory properties compared to the melissa leaf extract extracted by other extraction methods. It is effective in preventing and treating bowel disease, Alzheimer's, periodontitis, non-alcoholic fatty liver, and endometriosis. Therefore, the composition comprising the extract fraction of Melissa leaf of the present invention as an active ingredient is obesity, age-related macular degeneration, non-alcoholic steatohepatitis, psoriasis, cancer, diabetic retinopathy, glaucoma, Sjogren's syndrome, arteriosclerosis, arthritis, inflammatory bowel disease, It can be usefully used for the prevention or treatment of Alzheimer's, periodontitis, non-alcoholic fatty liver, and endometriosis, and has remarkably low side effects and excellent pharmacological effects.

Specifically, in the case of a pharmaceutical composition for the prevention or treatment of obesity comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it exhibits remarkable weight suppression, blood triglyceride suppression, blood sugar suppression, and total cholesterol suppression effect to prevent or treat obesity. There is an effect that can be.

In the case of a pharmaceutical composition for preventing or treating age-related macular degeneration comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it exhibits an effect of suppressing the size of macular degeneration lesions, thereby preventing or treating age-related macular degeneration. .

In the case of a pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it has a remarkable fibrosis inhibitory effect, suppresses the protein expression levels of Col1A2, TGF beta, IL-6 and IL-0 It has an effect that can prevent or treat non-alcoholic steatohepatitis.

In the case of a pharmaceutical composition for the prevention or treatment of psoriasis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it is possible to prevent or treat psoriasis by suppressing remarkable epidermal thickness, inflammatory cell infiltration of ear tissue, and mRNA expression level of ear tissue. There is an effect.

In the case of a pharmaceutical composition for preventing or treating cancer comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the size of the cancer tumor is significantly reduced, thereby preventing or treating cancer.

In the case of a pharmaceutical composition for preventing or treating diabetic retinopathy comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it has an effect of preventing or treating diabetic retinopathy because it has an excellent inhibitory effect on retinal vessel leakage.

In the case of a pharmaceutical composition for preventing or treating glaucoma comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, an increase in intraocular pressure is significantly suppressed, thereby preventing or treating glaucoma.

In the case of a pharmaceutical composition for preventing or treating Sjogren's syndrome comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the amount of saliva secretion and the amount of tear secretion increase, thereby preventing or treating Sjogren's syndrome.

In the case of a pharmaceutical composition for preventing or treating arteriosclerosis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the increase in atherosclerotic plaque is significantly suppressed, thereby preventing or treating arteriosclerosis.

In the case of a pharmaceutical composition for preventing or treating arthritis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it has an excellent arthritis treatment effect and an excellent weight-bearing increase effect, thereby preventing or treating arthritis.

In the case of a pharmaceutical composition for preventing or treating inflammatory bowel disease comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it has an effect of preventing or treating inflammatory bowel disease because it exhibits a low disease activity index.

In the case of a pharmaceutical composition for the prevention or treatment of Alzheimer, comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, cognitive function is improved, thereby preventing or treating Alzheimer's.

In the case of a pharmaceutical composition for the prevention or treatment of periodontitis comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the expression of pro-collagen increases, thereby effectively regenerating the gum tissue, thereby having a significant therapeutic effect on periodontitis. There is.

In the case of the pharmaceutical composition for the prevention or treatment of non-alcoholic fatty liver comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, it inhibits fat accumulation in hepatocytes, inhibits blood triglycerides, inhibits blood sugar, and inhibits total cholesterol. There is an effect that can be prevented or cured.

In the case of the pharmaceutical composition comprising the extract fraction of Melissa leaf according to the present invention as an active ingredient, the size of the uterine lesion is significantly reduced, so that endometriosis can be prevented or treated.

1 is a graph showing the change in body weight of obese rats induced by a high fat diet.
2 is a graph showing changes in feed intake of obese rats induced by high fat diet.
3 is a graph showing the change in the concentration of triglyceride in the blood of obese rats induced by a high fat diet.
4 is a graph showing changes in blood sugar concentration in obese rats induced by a high fat diet.
5 is a graph showing changes in blood total cholesterol concentration of obese rats induced by high fat diet.
6 is a graph showing the size of macular degeneration lesions in a laser-induced CNV model.
7 is a result of picrosirius red staining to detect fibrosis.
8 is a graph showing SMA deposition in the liver.
9 is a graph showing the protein expression level of Col1A2, which is known as a protein associated with liver fibrosis.
10 is a graph showing the protein expression level of TGF beta known as a protein associated with liver fibrosis.
11 is a graph showing the protein expression level of IL-6 known as a protein associated with liver fibrosis.
12 is a graph showing the protein expression level of IL-10 known as a protein associated with liver fibrosis.
13 is a graph showing the epidermal thickness of the extracted ear tissue.
14 is a graph showing the growth curve of tumor size in nude mice injected with DLD1 cells.
FIG. 15 is a result of testing the effect of retinal blood vessel leakage in a rat model of diabetic retinopathy induced by streptozotocin.
Figure 16 is a graph showing the intraocular pressure-lowering effect of the extract fraction of Melissa leaf according to the embodiment in the strut net scar glaucoma model induced by hypertonic saline.
17 is a graph analyzing the amount of tear secretion when the extract fraction of Melissa leaf according to the Example was administered using NOD/shilt, which is an animal model of Sjogren's syndrome.
18 is a graph showing the results of measuring and evaluating the formation of atherosclerotic plaques in the aorta in an ApoE deficient mouse model.
19 is a graph comparing the efficacy of arthritis treatment in a collagen-induced arthritis (CIA) induction model.
20 is a graph measuring the weight-bearing rate in the osteoarthritis model induced by MIA.
21 is a graph measuring the disease activity index (DIA) in an animal model of inflammatory bowel disease induced by dextran sodium sulfate (DSS).
22 is a graph showing a reference memory during an underwater maze test when a fraction of a Melissa leaf extract according to an example is administered.
23 is a graph showing the improvement rate of the probe test during the underwater maze test when the extract fraction of Melissa leaf extract according to the embodiment is administered.
FIG. 24 is a graph of RT-PCR results analyzing the gene expression level of pro-collagen indicating tissue regeneration in a periodontitis model induced by ligation.
25 is a graph showing the concentration values of AST and ALT in liver tissue.
26 is a graph showing the quantification of fat accumulation in liver tissue.
27 is a graph confirming the volume of uterine lesions in the experimental model of the experimental group to which the extract fraction of Melissa leaf extract was administered and the control group to which the extract of Melissa leaf extract was not administered.

Terms used in the present specification have selected general terms that are currently widely used as possible while taking functions of the present invention into consideration, but this may vary according to the intention or precedent of a technician working in the field, the emergence of new technologies, and the like. In addition, in certain cases, there are terms arbitrarily selected by the applicant, and in this case, the meaning of the terms will be described in detail in the description of the corresponding invention. Therefore, the terms used in the present invention should be defined based on the meaning of the term and the overall contents of the present invention, not a simple name of the term.

Unless otherwise defined, all terms used herein including technical or scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms such as those defined in a commonly used dictionary should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and should not be interpreted as an ideal or excessively formal meaning unless explicitly defined in the present application. Does not.

The numerical range includes the numerical values defined in the above range. All maximum numerical limits given throughout this specification include all lower numerical limits as if the lower numerical limits were expressly written. All minimum numerical limits given throughout this specification include all higher numerical limits as if the higher numerical limits were expressly written. All numerical limits given throughout this specification will include all better numerical ranges within the wider numerical range, as if the narrower numerical limits were expressly written.

In order to help understand the present invention, examples and manufacturing examples are presented. The following examples and preparation examples are provided for easier understanding of the present invention, and the contents of the present invention are not limited by the examples and preparation examples.

Example

Example 1. Preparation of melissa leaf extract fraction

After reflux extraction of 400.4 kg of dried Melissa leaves (origin: Europe) at 81° C. for 4 hours using 4000 L of 75% (v/v) ethanol, after 35 minutes of reflux extraction was completed, 10 μm of Filtered through a cartridge filter for 55 minutes to obtain 3100 L of the first extract.

The remnant of Melissa leaf was refluxed for 4 hours at 82°C using 4000 L of 75 (v/v)% ethanol, and then 35 minutes had elapsed after the reflux extraction was completed, and then 55 to 65 minutes with a cartridge filter of 10 μm. During filtration, 4100 L of a second extract was obtained.

After mixing the first and second extracts (a total of 7200 L of extract), concentration was performed to obtain 600 L of a first concentrate.

<First concentration condition>

Temperature: 56~58 ℃

Pressure: -0.066 ~ -0.070 MPa

Time: 9 hours

600 L of the first concentrate was concentrated to obtain 250 L of a second concentrate.

<Second concentration conditions>

Temperature: 56~58 ℃

Pressure: -0.063 to -0.065 MPa

Time: 5 hours 50 minutes

250 L of the second concentrate and 200 L of purified water were mixed to obtain a suspension (total of 450 L), and 450 L of ethyl acetate was added to the suspension to obtain 900 L of a mixed solution.

900 L of the mixed solution was stirred for 1 hour, and then allowed to stand for 2 hours to separate the ethyl acetate layer (450 L) to obtain a first fraction.

Thereafter, 450 L of ethyl acetate (the ratio of the mixture and ethyl acetate (v/v) = 1: 1) was added to the remaining suspension, followed by stirring for 1 hour, and then allowed to stand for 2 hours to obtain an ethyl acetate layer (450 L). Separated to obtain a second fraction.

The first fraction and the second fraction were combined (900 L) and concentrated to obtain 41 L (42.5 kg) of a third concentrate.

<Third concentration condition>

Temperature: 58 ℃

Pressure: -0.064 MPa

Time: 3 hours 40 minutes

The third concentrate was collected in a stainless X collection container for 20 minutes (42.5 kg).

Thereafter, after complete drying at 80∼82℃ for 36 hours using a hot air dryer (19.1 kg, 44.94% (w/w) of the concentrate), the dried product was put in a grinder and pulverized for 3 hours 50 minutes, and then the ethanol extract of Melissa leaf The ethyl acetate fraction of melissa leaf extract fraction was obtained (yield amount: 18.9 kg).

Example 2. Components of the extract fraction of melissa leaf

(1) Analysis method

The analysis method of the Melissa leaf extract was liquid chromatography (HPLC), and after injecting 10 l of the ethyl acetate extract sample prepared above into a Zorbax Eclipse Plus C18 (150 Х 4.6 mm) column, mobile phase A (5% Formic acid aqueous solution) and mobile phase B (methanol) were operated at a rate of 1 ml per minute according to the ratios shown in Table 1 below to analyze the active ingredients at a wavelength of 285 nm.

(2) result

The Melissa leaf extract fraction obtained according to Example 1 according to the above analysis method contained caffeic acid, EDPA, RME, and rosmarinic acid, and the contents thereof were as follows.

Caffeic acid: 0.96% by weight

EDPA: 0.71% by weight

RME: 0.11% by weight

Rosmarinic acid: 16.1% by weight

In addition, rutin was shown in the HPLC results of the extract fraction of Melissa leaf obtained according to Example 1. Therefore, it was found that the extract fraction obtained according to Example 1 did not contain rutin.

Experimental example

Experimental Example 1. Effect of inhibiting obesity in obese rats induced by high fat diet

(1) Experiment method

The effect of the extract fraction of Melissa leaf extract according to the examples in the treatment of obesity was evaluated in a high fat diet-induced obesity animal model.

Sprague-Dawley (SD) species in male rats (Purchased from: JoongAng Experimental Animal Co., Ltd.) in high-fat diet induced obese male rats ingested with high-fat feed (manufacturer: Saeron Bio Co., Ltd.) according to the embodiment Melissa The group to which the leaf extract fraction was administered was divided into an experimental group, and the group to which 0.5% carboxymethyulcellulose (CMC) was administered was divided into a control group.

The control group and the experimental group were adapted to the environment with a solid blended feed (purchased from: Saeron Bio Co., Ltd.) for 1 week before breeding with an experimental high fat diet, and then reared with an experimental diet for 10 weeks.

The administration method of the extract fraction of Melissa leaf was administered orally once a day, the dose was administered at 100 mg/kg, and 0.5% carboxymethyulcellulose (CMC) was administered to the control group. Body weight and feed intake were measured twice a week on the same day of the week. For the measurement of feed intake, a certain amount of feed was filled in a powder feed container at 3 pm, and their total weight was measured, and then the weight of the reduced feed was measured at 3 pm the next day. This was calculated as the amount of feed consumed for 24 hours, and it was carried out constantly until the end of the test. Blood samples were collected at the 5th and 10th weeks from the start of the test. Blood was collected by anesthesia with ether, and then a heparin-treated glass capillary tube was inserted into the orbital venous plexus of SD rats. Total cholesterol (total cholesterol) concentration, triglyceride (triglyceride) concentration and insulin (insulin) concentration were measured from the collected blood.

(2) result

1 is a graph showing the change in body weight of obese rats induced by a high fat diet. Referring to FIG. 1, in the case of the control group, continuous weight gain was observed, and this weight gain continued until the 10th week of the end of the test. However, in the case of the experimental group, the weight was significantly decreased in obese rats induced by high fat diet.

2 is a graph showing changes in feed intake of obese rats induced by high fat diet. 2, no significant change in feed intake was observed in the control group and the experimental group.

3 is a graph showing the change in the concentration of triglyceride in the blood of obese rats induced by a high fat diet. Referring to FIG. 3, in the case of the experimental group, blood triglycerides were significantly reduced in obese rats induced by a high fat diet. In particular, the amount of triglycerides in the blood did not increase in the experimental group even after ingesting a high fat diet for 10 weeks.

4 is a graph showing changes in blood sugar concentration in obese rats induced by a high fat diet. Referring to FIG. 4, in the case of the experimental group, blood sugar was significantly reduced in obese rats induced by a high fat diet.

5 is a graph showing changes in blood total cholesterol concentration of obese rats induced by high fat diet. Referring to FIG. 5, in the case of the experimental group, total cholesterol was significantly reduced in obese rats induced by high fat diet.

Experimental Example 2. Age-related macular degeneration inhibitory effect of the extract fraction of Melissa leaf in laser-induced CNV model

(1) Experiment method

The effect of treatment of age-related macular degeneration of the extract fraction of Melissa leaf according to the example was evaluated in a laser-induced choroidal neovascularization animal model.

A 6-week-old C57BL/6 mouse was damaged with a diode green laser (532 nm, 150 mW, 0.1 sec), and a laser-induced CNV model with a damaged retinal optic nerve was prepared.

The group to which the extract fraction of Melissa leaf according to the Example was administered to the laser-induced CNV model was divided into an experimental group, and the group to which 0.5% CMC was administered was divided into a control group.

The administration method of the extract fraction of Melissa leaf was administered orally once a day, the dose was 100 mg/kg, and was administered from two days before laser treatment, and 0.5% CMC was administered to the control group. The dosing period lasted 10 days after Lazer treatment.

After 10 days of laser irradiation, the retinal-choroid flat mount was performed to confirm the CNV lesion size inhibition effect. Mice were anesthetized and 25 mg/ml of FITC-dextran was injected retro-orbital. After 30 minutes, the mouse was euthanized, the eyeball was removed, fixed with 10% formalin, the cornea and lens were removed, and flat mounted on a cover glass. The size of the macular degenerative lesion size stained with FITC-dextran was measured.

(2) result

6 is a graph showing the size of macular degeneration lesions in a laser-induced CNV model. Referring to FIG. 6, it was shown that the size of macular degenerative lesions by laser irradiation in the control group increased, and in the case of the experimental group, the size of the neovascularization was significantly reduced in the laser-induced choroidal neovascularization model.

Experimental Example 3. Non-alcoholic steatohepatitis inhibitory effect of Melissa leaf extract fraction in a methionine/choline-deficient high fat diet model

(1) Experiment method

The effect of treatment of non-alcoholic steatohepatitis of the extract fraction of Melissa leaf according to the Example was evaluated in a methionine/choline-deficient high-fat diet animal model.

The study was conducted using C57Bl/6 mice fed methionine and choline-deficient high-fat diet (MCD-HFD), and methionine/choline-deficient high-fat dietary animal model according to the embodiment of Melissa leaf extract The group to which the fraction was administered was divided into an experimental group, and the group to which 0.5% CMC was administered was divided into a control group.

The administration method of the melissa leaf extract fraction was administered with the MCD-HFD feed and the melissa leaf extract fraction according to the example was orally administered once a day, the dose was 200 mg/kg, and the control group was administered 0.5% CMC. I did. The administration period proceeded for 9 weeks. In the case of the normal group, non-alcoholic steatohepatitis was not induced by supplying a normal diet.

In order to confirm the degree of fibrosis of the liver, picrosirius red staining was performed on the liver tissue. Sirius Red staining involves embedding a liver tissue sample fixed in 10% formalin in paraffin, cutting into 5 μm sections, incubating the tissue sections in 0.5% thiosemicarbazide for 10 minutes, and then saturating for 1 hour. Staining in 0.1% Sirius Red F3B in picric acid, then washed with acetic acid solution (0.5%). Immunohistochemical tests were performed using an antibody of Alpha-SMA (alpha-smooth muscle actin) to observe myofibroblast, which is an indicator of liver fibrosis. Paraffin-embedded liver tissue was made into a 4 μm section, attached to a slide glass treated with poly-L-lysine, deparaffinized, and then sequentially immersed in ethanol. To remove endogenous peroxidase, it was _{treated with 3% H 2} O ₂ /methanol solution for 10 minutes, and then reacted with normal goat serum for 40 minutes in a moisture chamber to remove non-specific reactions. , 0.05% fat-free milk powder and 0.01M phosphate-buffered saline solution containing 0.3% Triton X-100 (PBS, pH 7.4). The primary antibody was reacted for 1 hour each, and then reacted with the secondary antibody for 60 minutes, followed by color development. Western blot was performed to confirm the protein expression levels of Col1A2, TGF beta, IL-6, and IL-10. Specifically, the liver tissue was dissolved in a buffer containing a protease inhibitory cocktail, and the protein was extracted. The extracted protein was quantified using a BCA assay kit, mixed with SDS gel loading buffer, and boiled at 100°C for 5 minutes to denature. Protein electrophoresed on the SDS-PAGE gel was transferred to a nitrocellulose membrane. After that, to block non-specific protein binding, the membrane was left in 5% skim milk for 1 hour, and then Col1A2, TGF beta, IL-6, IL- A general immunoblot was performed by treatment with 10 and beta-actin, and then the intensity of the finally appeared band was measured and graphed with gel documentation software.

(2) result

7 is a result of Sirius Red staining to detect fibrosis. Referring to FIG. 7, fibrosis induced in a non-alcoholic steatohepatitis animal model induced by a high fat diet deficient in methionine/choline was inhibited in the experimental group to which the extract fraction of Melissa leaf according to the example was administered orally.

8 is a graph showing SMA deposition in the liver. Referring to FIG. 8, fibrosis induced in a non-alcoholic steatohepatitis animal model induced by a methionine/choline-deficient high-fat diet was suppressed in the experimental group to which the extract fraction of Melissa leaf according to the Example was administered orally.

9 to 12 are graphs showing protein expression levels of Col1A2, TGF beta, IL-6, and IL-10, which are known as proteins associated with liver fibrosis, respectively. 12 to 14, it was found that all of these proteins were increased in the control group, and the increased expression level of these proteins was suppressed by treatment (experimental group) with the extract fraction of Melissa leaf according to the Example.

Experimental Example 4. Psoriasis Inhibitory Effect of Fractions of Melissa Leaf Extract in IL-23 Induced Psoriasis Animal Model

(1) Experiment method

The effect of psoriasis treatment of the extract fraction of Melissa leaf according to the example was evaluated according to the method of Ma in an IL-23 induced psoriasis animal model [J. Clin. Cell Immunol. 4:6. (2013)].

An IL-23-induced psoriasis animal model was prepared by intradermally administering 500 ng of IL-23 to the ears of C57BL/6 mice (6 weeks old, Orient Bio) every other day for 16 days.

The group administered with the extract fraction of Melissa leaf according to the example to the IL-23-induced psoriasis animal model was classified as an experimental group, the group administered with 0.5% CMC was classified as a control group, and the control group treated with PBS instead of IL-23 was a PBS control group. It was classified as (PBS control).

As for the administration method of the melissa leaf extract fraction, the melissa leaf extract fraction according to the example was orally administered once a day, the dose was 100 mg/kg, and 0.5% CMC was administered to the control group. The administration period was orally administered from the day before IL-23 injection and proceeded for a total of 16 days, and the experiment was terminated on the 17th day of administration.

After the experiment was completed, the ear thickness of the mouse was measured, and the results are shown in FIG. 13.

(2) result

13 is a graph showing the epidermal thickness of the extracted ear tissue. 13, in the case of the experimental group, the epidermal thickness was significantly reduced in the IL-23 induced psoriasis animal model.

Experimental Example 5. Cancer Growth Inhibitory Effect of Melisa Leaf Extract Fraction in Tumor Formation Model Using Nude Mouse

(1) Experiment method

The anticancer effect of the extract fraction of Melissa leaf according to the example was evaluated in a tumor formation model using nude mice.

Colon cancer (SW480 and DLD1) cells were cultured in RPMI-1640 medium, and 6-week-old male nude mice (Orient Bio) were managed in a sterile facility. Cultured DLD1 cells were resuspended in DMEM medium, and 5 × 10 ⁶ cells were injected subcutaneously into the right flank of a nude mouse, and the group to which the Melissa leaf extract fraction according to the Example was administered was divided into an experimental group, and 0.5% The group to which CMC was administered was divided into a control group and tested.

As for the administration method of the melissa leaf extract fraction, the melissa leaf extract fraction was orally administered once a day for 28 days from the day of tumor injection, the dose was 100 mg/kg, and 0.5% CMC was administered to the control group. The administration period was administered for 28 days, and the tumor size was measured every 3 days with a caliper.

(2) result

14 is a graph showing the growth curve of tumor size in nude mice injected with DLD1 cells. In the control rats, visible tumors showing exponential growth were observed until the 28th day, but in the case of the experimental group to which the extract fraction of Melissa leaf extract according to the example was administered orally, the tumor was significantly reduced in size and showed a strong anticancer effect.

Experimental Example 6. The inhibitory effect of the extract fraction of Melissa leaf extract in a rat model of diabetic retinopathy induced by streptozotocin (STZ) on diabetic retinopathy

(1) Experiment method

The effect of the extract fraction of melissa leaf according to the example on the treatment of diabetic retinopathy was evaluated in a rat model of diabetic retinopathy induced by streptozotocin.

Using a citric acid buffer (100 mM, pH 4.5), a streptozotocin solution (100 mM) was injected into the abdominal cavity of rats at 150 mg/kg, and 10% sucrose was sufficiently supplied to prevent hypoglycemic shock. . Blood glucose was measured using a blood glucose meter from 2 days later, and a diabetic animal model in which non-fasting blood glucose was maintained at 300 mg/dl or more within 1 to 2 weeks was used.

Among the diabetes-induced rats, 0.5% CMC was administered to the control group, and the Melissa leaf extract fraction according to the example was administered to the experimental group, and the normal group did not induce diabetes.

The administration method of the melissa leaf extract fraction was to select the diabetes-induced rat and administer the streptozotocin solution (100 mM), and the melissa leaf extract fraction according to the example was orally administered once a day for 16 weeks from 2 weeks later. The dose was 100 mg/kg, and 0.5% CMC was administered to the control group. In order to quantitatively analyze the level of retinal leakage after 16 weeks, 1.25 mg of 500-kDa FITC-dextran (SigmaAldrich) was injected into the left ventricle of the rat, and blood was circulated for 5 minutes to stain. The eyeball was removed and immediately 4% para Fix for 45 minutes using formaldehyde. The retina was cut from the fixed eyeball, cut in a Maltese cross shape, and the cut retina was placed on a slide glass, and then observed using a confocal microscope. The level of retinal leakage was measured by measuring the intensity of FITC-dextran exuding from the entire retinal tissue.

(2) result

FIG. 15 is a result of testing the effect of retinal blood vessel leakage in a rat model of diabetic retinopathy induced by streptozotocin. Referring to FIG. 15, as a result of testing the effect of inhibiting retinal vessel leakage, it was confirmed that the degree of leakage of retinal vessels was high in the control group. Significantly inhibited the leakage of retinal blood vessels by significantly inhibiting the leakage.

Experimental Example 7. Inhibitory effect of melissa leaf extract fraction in glaucoma model induced by hypertonic saline solution

(1) Experiment method

The glaucoma treatment effect of the extract fraction of Melissa leaf according to the example was evaluated in a strut net scar glaucoma model induced by hypertonic saline.

Rats were anesthetized by intraperitoneal injection of ketamine-xylazine, and 50 μl of 1.8 M hypertonic saline was injected into the episcleral veins in the left eye. scar) induces an increase in intraocular pressure due to the resistance of the ocular discharge path. The right eyeball was used as the control site, and the intraocular pressures of the left and right eyes were measured for 2 weeks before and at 2 days intervals after glaucoma induction.

After selecting the glaucoma-induced rats, 0.5% CMC was administered to the control group, and the melissa leaf extract fraction according to the example was administered to the experimental group, and glaucoma was not induced in the normal group.

The administration method of the experimental group was administered orally once a day with the extract fraction of Melissa leaf according to the example for 2 weeks from the date of induction of glaucoma in order to test the effect of the test substance, the dose was 100 mg/kg, and the control group was 0.5%. CMC was administered.

Figure 16 is a graph showing the intraocular pressure-lowering effect of the extract fraction of Melissa leaf according to the embodiment in the strut net scar glaucoma model induced by hypertonic saline. Referring to FIG. 16, in the case of the control group, it was confirmed that the intraocular pressure reached the highest value and continued to be maintained on the 6th day after the hypertonic saline injection, and in the case of the experimental group, the intraocular pressure increased in the strut net scar glaucoma model induced by hypertonic saline. Significantly alleviated.

Experimental Example 8. Efficacy evaluation of the extract fraction of Melissa leaf using Sjogren's syndrome animal model

(1) Experiment method

Experiments were performed using NOD/shilt mice, which is an animal model of Sjogren's syndrome.

In NOD/shilt, which is an animal model of Sjogren's syndrome, the control group was administered 0.5% CMC, and the experimental group was administered the extract fraction of Melissa leaf extract according to the example.

As for the administration method of the melissa leaf extract fraction, the amount of tear secretion of the mouse was analyzed after administering the melissa leaf extract fraction according to the example at 100 mg/kg each for 12 weeks. After anesthetizing NOD/shilt mice with isoflurane, tear secretion using a cotton thread soaked with phenol red according to Zoukhri et al. (Exp Eye Res. 2007; 84:894-904) Was measured.

(2) result

17 is a graph analyzing the amount of tear secretion when the extract fraction of Melissa leaf according to the Example was administered using NOD/shilt mouse, which is an animal model of Sjogren's syndrome. Referring to FIG. 17, in the case of the experimental group, the amount of tear secretion was significantly increased.

Through the above contents, it was confirmed that the amount of saliva secretion and the amount of tear secretion increased by administering the extract fraction of Melissa leaf extract according to the Example, thereby improving Sjogren's syndrome.

Experimental Example 9. Inhibitory effect of extract fraction of Melissa leaf extract in apolipoprotein E (ApoE) deficient mouse model on arteriosclerosis

(1) Experiment method

The effect of the extract fraction of Melissa leaf extract according to the example in the treatment of arteriosclerosis was evaluated in a mouse model deficient in ApoE. Mice that induced arteriosclerosis through a high-fat diet for 16 weeks were selected, and those with no arteriosclerosis were classified as a normal group by proceeding with a free diet instead of a high-fat diet.

0.5% CMC was administered to the control group, and the Melissa leaf extract fraction according to the example was administered to the experimental group.

Diet and water were fed freely for 16 weeks, and the extract fraction of Melissa leaf according to the example was orally administered once a day at 100 mg/kg to rat mice that induced arteriosclerosis through a high fat diet for 16 weeks. 0.5% CMC was administered.

After 16 weeks, the mice were sacrificed and resected from the heart and the ascending aorta to the thoracic aorta and fixed in a 10% neutral buffered formalin solution. After trimming the fixed heart with a razor, embedding it in a frozen tissue embedding agent (OCT compound) and freezing it in a deep freezer, the aortic arch is 10 μm with a frozen section. Cut to thickness to prepare a slide. For fat dyeing, the slides were immersed in distilled water, treated with propylene glycol for 1 minute, dyed in an oil-red solution for 16 hours, and then treated in 85% propylene glycol for 2 minutes. After washing the slide with distilled water, it was encapsulated with an aqueous encapsulant and observed with an optical microscope.

(2) result

18 is a graph showing the results of measuring and evaluating the formation of atherosclerotic plaques in the aorta in an ApoE deficient mouse model. Compared to the normal group, it was confirmed that the formation of atherosclerotic plaque was increased in the control group, and in the case of the experimental group, the increase in atherosclerotic plaque was significantly suppressed.

Experimental Example 10. Effect of arthritis treatment in an animal model of collagen-induced arthritis

The effect of the extract fraction of Melissa leaf extract according to the example in the treatment of arthritis was evaluated in a collagen-induced arthritis model.

The collagen induced arthritis (CIA) animal model was prepared by inducing an immune response by injecting collagen, which is believed to be the cause of rheumatoid arthritis.

4 mg of chicken type II collagen was mixed with 1 ml of 100 mM acetic acid and dissolved at 4° C. for one day. The mixture was emulsified with 1 ml of Freund's complete adjuvant containing 4 mg/ml of Mycobacterium tuberculosis, and then 150 μl (300 μg) was injected intradermally into the tail of a 6-week-old Lewis rat to immunize it. On the 7th day after the primary immunization, the chicken's type II collagen was injected intradermally to induce a secondary immune response, and arthritis symptoms could be confirmed 1 to 2 weeks after the secondary immunization.

The control group was administered 0.5% CMC, and the experimental group was administered the extract fraction of Melissa leaf according to the example.

For 47 days from the next day after induction of the secondary immune response, the extract fraction of Melissa leaf according to the example was orally administered once a day, the dose was 100 mg/kg, and 0.5% CMC was administered to the control group.

In order to verify the treatment effect, the degree of swelling and erythema of joints and nodes were observed for each rat's foot, and the clinical score was calculated according to the scoring index shown in the table below.

(2) result

19 is a graph comparing the efficacy of arthritis treatment in a collagen-induced arthritis (CIA) induction model. Referring to FIG. 19, arthritis symptoms appeared from about 20 days after the CIA-induced immune response. In the case of the experimental group, the collagen-induced arthritis induction model showed a significant treatment effect.

Experimental Example 11. Effect of treatment on osteoarthritis in an animal model of osteoarthritis induced by MIA (Monosodium iodoacetate)

(1) Experiment method

The effect of the extract fraction of Melissa leaf extract according to the Examples in the treatment of osteoarthritis was evaluated in a model of osteoarthritis induced by MIA.

Osteoarthritis inducing substance MIA (diluted at a concentration of 60 mg/ml with 0.9% saline) was administered at a rate of 50 μl into the joint cavity of the right hind limb of 7-week-old SD rats to induce osteoarthritis and selected.

0.5% CMC was administered to the control group, and the Melissa leaf extract fraction according to the example was administered to the experimental group.

The administration method of the experimental group was administered orally once a day with the extract fraction of Melissa leaf according to the example for 21 days, the dose was 100 mg/kg, the control group was administered 0.5% CMC, and body weight at 7 days intervals after administration. The load factor was measured. The group that did not induce osteoarthritis was classified as a normal group.

The hind limb weight load was measured using a foot weigher. In the holder of the tester, the rat with osteoarthritis induces strength by relying on a normal foot without MIA due to pain, so the weight of both feet was out of balance and the weight of the foot administered with MIA was relatively light compared to the weight of the normal foot. When measuring the weight of the feet, the weight (g) of both feet was measured in the state that the stomach of the SD rat did not touch the sensor of the device.

Using the measured weight of the foot, the weight-bearing rate (%) was calculated by the following equation.

[Equation 1]

Weight-bearing rate (%) = [weight of hind limbs causing osteoarthritis/(weight of hind limbs of both feet)] × 100

20 is a graph measuring the weight-bearing rate in the osteoarthritis model induced by MIA. Referring to FIG. 20, it was confirmed that the weight-bearing ratio (%) of the control group compared to the normal group was statistically significantly decreased compared to the normal group as the period elapsed.

In contrast, in the case of the experimental group, the decreased weight-bearing rate was significantly increased, thereby showing an excellent effect of increasing weight-bearing (osteoarthritis treatment effect).

Experimental Example 12. Inhibitory effect of inflammatory bowel disease in an inflammatory bowel disease model induced by dextran sodium sulfate (DSS)

The effect of treatment of ulcerative colitis of the extract fraction of Melissa leaf according to the example was evaluated in an animal model induced by sodium dextran sulfate.

7-week-old C57BL/6J mice (central laboratory animals) were acclimated for 1 week while freely supplied with feed and water, and then randomly divided into a control group, a normal group, and an experimental group.

For 11 days after group separation, the Melissa leaf extract fraction according to the example was orally administered once a day, the dose was 200 mg/kg, and 0.5% CMC was administered to the control group. The induction of inflammatory bowel disease was induced by mixing water containing 3% DSS in drinking water instead of water and ingesting it freely from the 3rd day of group separation. On the other hand, in the case of the normal group, DSS was not administered and did not cause inflammatory bowel disease.

During the experiment period, the appearance of the stool was observed by inducing a bowel movement of the mouse once a day, and the presence or absence of occult blood was observed using coulter hemoccult single slides, and scored according to the criteria in Table 3 below.

[Equation 2]

DAI (Disease Activity Index) = stool concentration + stool color 21 is a graph measuring the disease activity index (DIA) in an inflammatory bowel disease animal model caused by sodium dextran sulfate (DSS). At this time, the DIA of the normal group represents 0. As a result of measuring DAI in rats with ulcerative colitis induced by sodium dextran sulfate (DSS), the control group showed a high DAI, whereas the experimental group showed a significantly low DAI level, showing a remarkable therapeutic effect.

Experimental Example 13. The effect of improving cognitive function in an animal model of Scopolamine-induced dementia

For the cognitive function improvement test, 9-week-old dominant ICR mice were used for each treatment group. The group to which the extract fraction of Melissa leaf extract was administered was classified as an experimental group, and as a vehicle, a control group treated with scopolamine after administration of physiological saline instead of the drug for 2 weeks was used.

For the administration of the experimental group, 200 mg/kg was administered orally at a dose of 7 times a week for 2 weeks, and scopolamine was administered orally at 1 mg/kg 60 minutes before the start of the test to induce memory impairment. A water maze test was conducted.

During the underwater maze test, the reference memory test was conducted after acclimatization by allowing them to swim freely for 60 seconds in a tank without a platform one day before the start of the test. The reference memory test was performed by measuring the time (escape latency, unit: seconds) to go up to find a platform submerged in water 4 to 5 times a day for 5 days, and the maximum allowable time was 60 seconds. Limited. If the platform was not located until the second day of the test, the platform was guided to find the platform within the time limit of 60 seconds.

24 hours after the end of the reference memory test, the platform in the tank was removed and allowed to swim freely for 60 seconds to measure the time to stay at the position where the platform was, and a probe test was performed.

After administration of the extract fraction of Melissa leaf according to the example, a reference memory test was conducted for 5 days, and the average value of the time to find the platform of the scopolamine administration group on the 5th day and the time to find the platform of each sample were compared with the administration of scopolamine. The time to find the platform compared to one control group was compared.

(2) result

Referring to FIG. 22, when the extract fraction of Melissa leaf according to Example 1 was administered, the effect of improving cognitive function was confirmed (37.2±2.1).

In the probe test conducted after the end of the reference memory test, the average time of staying at the site of the platform in the scopolamine-administered group was administered with the fraction of Melissa leaf extract according to the example, and compared with the average time of staying, it was expressed as an improvement rate.

[Equation 3]

Probe test improvement rate = (tA-ts)/tsХ 100(%)

The ts is the average time of staying at the site where the scopolamine administration group was at the platform, and tA is the average time at which the administration group of the Melissa leaf extract fraction according to the embodiment stayed at the site of the platform.

Referring to FIG. 23, even in the probe test, the experimental group had a longer stay at the platform than the control group, and the experimental group (88.3±9.9%) showed an excellent improvement rate compared to the control group.

Experimental Example 14. Inhibitory effect of periodontitis in a periodontitis model induced by ligation

(1) Experiment method

The experimental animals were 7-week-old male SD rats, and they were fully supplied with general laboratory animal feed and water, and were used for the experiment after 1 week of acclimatization. To induce periodontitis, periodontitis was induced by ligating the mandibular first molar with a sterilized suture (3-0, nylon thread) after general anesthesia, and rats without ligation were classified as normal after checking only the tooth neck.

Of the group in which periodontitis was induced, the extract fraction of Melissa leaf according to the Example was administered to the experimental group.

The administration route of the experimental group was orally administered once a day, the dose was 100 mg/kg, and 0.5% CMC was administered to the control group.

After the sacrifice of the experimental animal, the surrounding tissues of the mandible were removed, and then the gene expression level of procollagen indicating tissue regeneration was confirmed.

In periodontal disease, regeneration and recovery of periodontal tissue made of collagen is important. The gene expression level of procollagen from the gums was confirmed through RT-PCR.

10 μg of RNA obtained by crushing the extracted gum tissue with a homogenizer using the Trizol method was synthesized using RT-PCR (reverse transcriptase-polymerase chain reaction) to synthesize cDNA. The nucleotide sequence of the PCR primer for each gene is shown in Table 4 below.

(2) result

FIG. 24 is a graph of the results of RT-PCR analyzing the gene expression level of pro-collagen indicating tissue regeneration in a periodontitis model induced by ligation. Referring to FIG. 30, it can be seen that the administration of the extract fraction of Melissa leaf according to the Example increases the expression of pro-collagen to effectively regenerate the gum tissue, thereby showing a significant therapeutic effect on periodontitis.

Experimental Example 15. Non-alcoholic fatty liver inhibitory effect of the extract fraction of Melissa leaf in a methionine/choline deficient high fat diet model

(1) Experiment method

As experimental animals, 30 SD rats were purified for 1 week and then randomly separated into three groups of 10 animals.

For each of the three classified groups, the group that supplied the solid compound feed (purchaser: Saeron Bio Co., Ltd.) was divided into the normal group, and the group supplied with a high fat diet (HFD) and administered the melissa leaf extract fraction of the example The experiment was divided into an experimental group, and the group fed a high fat diet (HFD) and administered 0.5% CMC was divided into a control group and tested.

The normal group was fed a solid blended feed. The experimental group was fed HFD feed instead of the solid compound feed, and at the same time as feeding the HFD feed, the Melissa leaf extract fraction according to the example was orally administered once a day, and the dose of the Melissa leaf extract fraction was 100 mg/kg. The control group was experimented in the same manner as the experimental group, except that 0.5% CMC was administered instead of the extract of Melissa leaf of the example. The administration period proceeded for 10 weeks.

In order to measure liver values (AST, ALT) of each of the normal group, the experimental group, and the control group, rats were autopsied to extract a blood sample from the abdominal aorta. The extracted blood sample was centrifuged to obtain serum. Serum liver values (AST, ALT) were measured using an automatic biochemical analyzer.

Three rats were randomly selected from each of the normal group, the experimental group, and the control group to check the degree of fat accumulation in the liver tissue of the normal group, the experimental group, and the control group. Four stained slides stained with oil red on the frozen liver tissue of the selected rat were prepared for each individual, and then the degree of fat accumulation in the liver tissue was quantified using an analysis program (ImageJ).

(2) result

25 is a value (unit: IU/L) obtained by measuring liver values (AST, ALT) of each of the normal group, the experimental group, and the control group. Referring to FIG. 25, in the case of the normal group, it was confirmed that fatty liver did not occur as a bar showing low liver values (AST, ALT). The experimental group to which the extract fraction of Melissa leaf extract according to the embodiment was administered showed lower liver values (AST, ALT) than the control group, and it was confirmed that fatty liver was suppressed compared to the control group.

26 is a graph showing the degree of fat accumulation of each of the experimental group and the control group by quantifying it with an analysis program (ImageJ). Referring to Figure 26, in the case of the normal group, it is confirmed that the degree of accumulation of fat in the liver tissue is insignificant. It was confirmed that the experimental group administered with the extract fraction of Melissa leaf extract according to the Example had suppressed the degree of fat accumulation in the liver tissue compared to the control group administered with CMC.

Experimental Example 16. Endometrial growth inhibitory effect of the extract fraction of Melissa leaf in the endometrial autograft model

(1) Experiment method

After wearing the SD rat, in order to match the female's estrus cycle, a bedding moistened with male urine was placed in the female cage at a 5-day cycle, and vaginal cytology was performed daily for a week before surgery to confirm the menstrual cycle.

Before surgery, the surgical site was shaved with an electric razor and the surgical site was disinfected with 70% ethanol. The left uterus was exposed after an approximately 1 cm incision was made at a location of 0.5 cm to 1.0 cm in the vaginal opening. Both parts of the exposed left uterus at 6 cm to 8 cm positions were tightly tied with 5-0 sutures. Cut the uterus between the tied positions, place it in a sterile glass Petri dish containing 100 μL of PBS containing penicillin (100 U/ml) and streptomycin (100 μg/ml), remove the fat, and incise the uterus vertically into 3 pieces of 2 mm size. I cut it out.

After finding the intestine from the incision site, pulling it out, spreading it on pre-soaked gauze, checking the three arteries between the mesentery, and gently suturing the uterine piece. Then, the intestine was put into the abdominal cavity, and the abdominal wall was sewn using sutures, and the skin was closed with an autoclip. After the operation, heat was applied to prepare SD rats to lower their body temperature. The surgical site was disinfected with povidone, and Acetaminophen, an analgesic drug, was mixed with drinking water at 2 mg/ml and fed.

The auto clip was removed 7 days after the operation, and the extract fraction of Melissa leaf according to the example was orally administered once a day for 4 weeks to the experimental group (n = 5). The dosage of the extract fraction of melissa leaf was 100 mg/kg. Group separation was performed so that the average size of uterine lesions in each group was the same.

To the control group (n = 5), 0.5% of CMC was orally administered.

Four weeks after surgery, SD rats were euthanized and the peritoneum was incised. Thereafter, the uterine lesion implanted in the mesentery was isolated and the size and weight were measured.

(2) result

27 is a graph confirming the volume of uterine lesion tissue in the experimental model of the experimental group to which the extract fraction of Melissa leaf extract was administered and the control group to which the extract of Melissa leaf extract was not administered. In Fig. 27, the average value of the volume of uterine lesion tissues of the experimental group and the control group is described.

Table 5 is a table describing the size and volume of uterine lesions in each of the experimental and normal groups.

Width(mm) Length(mm) Volume (mm ³ ) Example SD Rat 1 1.82 1.51 2.16 Example SD Rat 2 1.75 1.53 2.13 Example SD Rat 3 1.59 1.39 1.60 Example SD Rat 4 1.63 1.28 1.39 Example SD Rat 5 1.99 1.16 1.39 Comparative Example SD Rat 1 2.56 2.01 5.38 Comparative Example SD Rat 2 2.39 2.06 5.27 Comparative Example SD Rat 3 2.47 1.98 5.04 Comparative Example SD Rat 4 2.27 1.82 3.91 Comparative Example SD Rat 5 2.45 1.89 4.55

Referring to Table 5, the average volume of uterine tissue in the experimental group is about 1.73 mm ³ and the standard deviation is about 0.38. The average volume of uterine tissue of the control group is about 4.83 mm ³ and the standard deviation is about 0.61. Referring to Figs. 27 and 5, it is confirmed that the volume of the uterine tissue of the experimental group is significantly smaller than the volume of the uterine tissue of the control group . Through this, it can be seen that the extract of Melissa leaf of the embodiment has an excellent effect of inhibiting the growth of endometrial tissue.

Claims (27)

As a pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis using the extract of the melissa leaf fraction as an active ingredient, wherein the extract of the melissa leaf fraction comprises the step of extracting and concentrating the melissa leaf with 50% to 100% alcohol, and then fractionating. A pharmaceutical composition comprising 0.1 to 5% by weight of EDPA (Ethyl 2-(3,4-dihydroxyphenyl) acetate) obtained by an extraction process, wherein the extract of the Melissa leaf fraction comprises 0.1 to 5% by weight of EDPA (Ethyl 2-(3,4-dihydroxyphenyl) acetate). The pharmaceutical according to claim 1, wherein the extract of the Melissa leaf fraction comprises 0.1 to 5% by weight of caffeic acid, 0.1 to 5% by weight of EDPA, 0.01 to 2% by weight of RME, and 5 to 50% by weight of rosmarinic acid. Composition. delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete As a method for preparing the extract of the Melissa leaf fraction as defined in claim 1 or 2,
The first step of extracting the Melissa leaf with 50% to 100% alcohol;
A second step of concentrating the product of the first step; And
Suspending the product of the second step and fractionating it with ethyl acetate,
The Melissa leaf fraction extract is used as an active ingredient in a pharmaceutical composition for the prevention or treatment of non-alcoholic steatohepatitis. The method of claim 24, wherein the alcohol is 70 to 80% (v/v) of ethanol. The method of claim 24, wherein the second step
(1) a first concentration step of concentrating the alcohol extract for 5 to 15 hours in a temperature condition of 50 to 60° C. to obtain a first concentrate, and
(2) A method comprising a second concentration step of concentrating the first concentrate for 3 to 10 hours at a temperature of 55 to 60° C. to obtain a second concentrate. The method of claim 26, further comprising a third concentration step of concentrating the fractions for 3 to 10 hours under a temperature condition of 55 to 60 °C after performing the first concentration step and the second concentration step. The manufacturing method.

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