WO2011112002A2 - 들쭉 추출물 또는 들쭉 분획물을 유효 성분으로 함유하는 황반 변성증의 치료, 예방 또는 개선용 조성물 - Google Patents

들쭉 추출물 또는 들쭉 분획물을 유효 성분으로 함유하는 황반 변성증의 치료, 예방 또는 개선용 조성물 Download PDF

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WO2011112002A2
WO2011112002A2 PCT/KR2011/001631 KR2011001631W WO2011112002A2 WO 2011112002 A2 WO2011112002 A2 WO 2011112002A2 KR 2011001631 W KR2011001631 W KR 2011001631W WO 2011112002 A2 WO2011112002 A2 WO 2011112002A2
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fraction
blueberry
macular degeneration
extract
pharmaceutical composition
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PCT/KR2011/001631
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English (en)
French (fr)
Korean (ko)
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WO2011112002A3 (ko
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정세영
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주식회사 호일바이오메드
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Priority to CN201180016617.7A priority Critical patent/CN102821773B/zh
Priority to JP2012556977A priority patent/JP5860418B2/ja
Publication of WO2011112002A2 publication Critical patent/WO2011112002A2/ko
Publication of WO2011112002A3 publication Critical patent/WO2011112002A3/ko

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/45—Ericaceae or Vacciniaceae (Heath or Blueberry family), e.g. blueberry, cranberry or bilberry
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

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  • the present invention relates to a composition for the treatment, prevention or improvement of macular degeneration containing a blueberry (Vaccinium uliginosum) extract or a blueberry fraction as an active ingredient.
  • a blueberry Vaccinium uliginosum
  • macular degeneration the nerve tissue located in the central part of the inner retina of the eye. Most of the cells are gathered here, and the place where the image of the object is also the center of the macular and plays a very important role in vision. Eye disease that causes degeneration of the macula due to various causes and causes visual impairment is called macular degeneration (macular degeneration), and macular degeneration is known as one of the three major blindness diseases along with glaucoma and diabetic retinal disease.
  • This macular degeneration affects central vision, causing blurred vision in the center of vision, making it difficult to perform sophisticated activities such as reading and driving due to central dark spots, schizophrenia (a symptom of being distorted) or loss of vision in the local area. It is a very serious disease that, in severe cases, can lead to blindness.
  • Age-related Macular Degeneration has long been the most common disease causing blindness in older people over 55 in Western society, and in the United States alone, more than 8 million people are currently a form of senile macular degeneration. It has dry AMD, and more than 3 million people are reported to lose sight each year.
  • AMD Age-related Macular Degeneration
  • macular degeneration patients are increasing exponentially in Korea, which is expected to be caused by increased exposure to ultraviolet light due to ozone depletion in addition to westernized eating habits.
  • macular degeneration was originally a disease mainly occurring in the elderly, but in recent years, the age of the invention has been lowered from the elderly in their 60s to the 4 and 50s in the elderly, and the incidence of middle-aged people has also increased rapidly in recent years.
  • Macular degeneration can be classified into two types, dry and wet, and about 90% of people with macular degeneration are patients with dry macular degeneration.
  • Dry macular degeneration is caused by thinning or shrinking macular tissues, resulting in the accumulation of waste products to kill retinal cells, and wet macular degeneration causes damage to the macula due to rupture of new neoplasms due to the buildup of waste products in or below the retina. This happens.
  • the dry form it is characterized by difficulty in self-diagnosis because there is no sign at first, and it progresses slowly, and there is a relatively low risk of blindness, but it is possible to convert into wet macular degeneration with high risk of blindness. It is very important to prevent this disease.
  • Ranibizumab a humanized monoclonal antibody that recognizes and blocks vascular endothelial growth factor (VEGF A) and is known to be effective in restoring or maintaining vision in wet macular degeneration.
  • VEGF A vascular endothelial growth factor
  • ranibizumab may increase the risk of stroke, and ranibizumab is expensive and injectable in the eye.
  • the disadvantage is that it is inconvenient.
  • Lutein protects the macula of the retina and maintains the macular pigment density to prevent macular degeneration, but studies show that prolonged use of supplements containing carotenoids, such as lutein, increases the risk of developing lung cancer, especially for smokers Appeared. Therefore, it can be dangerous for smokers to take lutein-related products.
  • zeaxanthin which is present as macular pigment, such as lutein, and anthocyanin contained in berries.
  • zeaxanthin which is present as macular pigment, such as lutein, and anthocyanin contained in berries.
  • the present invention is to recognize and improve the problems with the existing treatment and prevention methods for macular degeneration as described above, more specifically, the present invention is mainly from the natural plant blueberry (Vaccinium uliginosum) used for food It is an object of the present invention to provide a composition for the treatment or prevention of new macular degeneration with excellent therapeutic and prophylactic effects without any side effects even after long-term use by using the extracted extract as a main component.
  • the natural plant blueberry Vaccinium uliginosum
  • the present invention is to provide a cosmetic composition, food composition and pharmaceutical composition for improving macular degeneration including blueberry extract.
  • the present invention provides a composition for improving or treating macular degeneration containing an extract derived from blueberry (Vaccinium uliginosum) as an active ingredient.
  • an extract derived from blueberry Vaccinium uliginosum
  • A2E pyridnium bis-retinoid
  • ultraviolet radiation increases the risk of macular degeneration or accelerates the progression of the disease, since radical changes in retinal pigment epithelial cells are dramatically increased when A2E is converted to A2E in the photooxidized form by ultraviolet radiation.
  • the present inventors have been studying to find a substance having inhibitory ability to accumulate A2E and A2E oxidation in retinal pigment epithelial cells for the fundamental treatment and prevention of macular degeneration, while extracts from blueberries inhibit A2E accumulation in retinal pigment epithelial cells. And inhibiting A2E oxidation by ultraviolet rays and found to have an effect of protecting retinal pigment epithelial cells, and scientifically proved this by experimental results to confirm that the treatment and prevention of macular degeneration can be confirmed. To complete.
  • Blueberry which is used for the first time as an ingredient for the treatment or improvement of macular degeneration, is a botanical and deciduous shrub that grows in Hallasan, Geumgangsan, Baekdusan, etc. on the Korean Peninsula, which blooms in June-July and bears fruit in August. .
  • the main components of blueberry include sugar (8-11.8%), fruit acid (2-2.25%), tannin (0.15-0.25%), fiber, and the like.
  • the blueberry which has been mainly used for food, is adopted as a raw material for improving, preventing or treating macular degeneration, and the composition for preventing or treating macular degeneration of the present invention is also suitable for oral use.
  • the composition for preventing or treating macular degeneration of the present invention is also suitable for oral use.
  • eye drops, eye ointments, etc. for the treatment or improvement of macular degeneration is very suitable for use directly to the eye.
  • the components extracted from blueberries using ARPE-19 cells have an effect of inhibiting A2E accumulation and inhibiting A2E oxidation, as described below.
  • the blueberry fraction can fundamentally cure the pathogenesis of macular degeneration, such as A2E accumulation and A2E oxidation, and can be very useful for the treatment, prevention and improvement of macular degeneration through a pharmaceutical composition or functional food containing the same as an active ingredient. have.
  • it can be usefully used for the treatment, prevention and improvement of senile macular degeneration.
  • the present invention provides a composition for treating, preventing or ameliorating macular degeneration, which contains blueberry extract or a fraction fractionated therefrom as an active ingredient.
  • the blueberry extract is extracted from the flowers, berries, leaves or bark of the blueberry.
  • the extraction method may be separated according to a conventional method known in the art for extracting natural products, that is, using a solvent conventionally used in the art under conditions of ordinary temperature, pressure, and time.
  • a solvent which can be generally used in the extraction process can be used, and it is preferable to extract using water, C 1-4 anhydrous or hydrous lower alcohol. It can be extracted using a solvent selected from the group consisting of acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol.
  • the blueberry fraction may be obtained from the flowers, fruits, leaves or bark of the blueberry, and may also be obtained from the blueberry extract.
  • the blueberry fraction which exhibits a particularly significant effect in the present invention is the polyphenol fraction of the blueberry, the pigment fraction of the blueberry or a mixture thereof, and preferably the polyphenol fraction of the blueberry.
  • the blueberry fraction may be fractionated by a fractionation method such as pigment fractionation, polyphenolic fractionation, etc., which are well known in the art, for example, solid phase extraction (SPE) as used in the present embodiment.
  • SPE solid phase extraction
  • the blueberry fraction can be separated according to the appropriate order by selecting an elution solvent suitable for the component to be separated, and a polycarbohydrate / acid fraction containing a large amount of carbohydrate / acid component and a polyphenolic compound (flavonol, etc.) It can be separated into a phenol fraction, a pigment fraction containing a large amount of pigment components (such as anthocyanin) and the like.
  • the elution solvent may be appropriately selected by those skilled in the art, and as used in this example, ethyl acetate may be used to obtain a polyphenol fraction, and HCl / methanol aqueous solution may be used to obtain a pigment fraction. Only the polyphenol fraction, the pigment fraction of the blueberry or a mixture thereof among the blueberry fractions show a significant effect, and the polyphenol fraction is most preferred.
  • the composition of the present invention may be provided in the form of a pharmaceutical composition comprising a pharmaceutically effective amount of a blueberry extract.
  • pharmaceutically effective amount means an amount sufficient for the above-mentioned effective ingredient compound to achieve the effect of treating or preventing macular degeneration.
  • the pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier in addition to the blueberry extract or blueberry fraction.
  • pharmaceutically acceptable carriers are those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.
  • the pharmaceutical composition of the present invention may include diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and other pharmaceutically acceptable additives.
  • Suitable dosages of the pharmaceutical compositions of the present invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and reaction sensitivity of the patient.
  • it can be administered in a dose of 0.001 to 1000 mg / kg per day, the administration can be administered once or several times a day.
  • the pharmaceutical composition of the present invention can be administered orally or parenterally.
  • the dosage form include eye drops, eye ointments, injections, tablets, capsules, granules, powders, and the like. These can be formulated using techniques that are widely used.
  • eye drops include isotonic agents such as sodium chloride and concentrated glycerin, buffering agents such as sodium phosphate and sodium acetate, surfactants such as polyoxyethylene sorbitan monooleate, polyoxyl stearic acid 40 and polyoxyethylene hydrogenated castor oil, and citric acid Stabilizers, such as sodium and sodium edetate, and preservatives, such as benzalkonium chloride and paraben, etc. can be used as needed.
  • pH should just be in the range which is acceptable for ophthalmic preparations, the range of 4-8 is preferable.
  • ophthalmic ointment can be prepared using a common base such as white petrolatum, liquid paraffin.
  • oral preparations such as tablets, capsules, granules, and powders include bulking agents such as lactose, crystalline cellulose, starch, vegetable oil, glidants such as magnesium stearate and talc, and binders such as hydroxypropyl cellulose and polyvinylpyrrolidone.
  • Disintegrating agents such as calcium carboxymethyl cellulose, low-substituted hydroxypropyl methyl cellulose, coating agents such as hydroxypropyl methyl cellulose, macrogol, silicone resins, and coating agents such as gelatin coatings, etc. Can be.
  • the compositions of the invention may be provided in the form of food compositions, in particular functional food compositions.
  • Functional food compositions of the present invention include ingredients that are commonly added in the manufacture of food, and include, for example, proteins, carbohydrates, fats, nutrients and seasonings.
  • natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.); Disaccharides (eg maltose, sucrose, etc.); oligosaccharide; Polysaccharides (eg, dextrins, cyclodextrins, etc.); And sugar alcohols (eg, xylitol, sorbitol, erythritol, and the like).
  • natural flavoring agents eg, taumartin, stevia extract, etc.
  • synthetic flavoring agents eg, saccharin, aspartame, etc.
  • the invention provides a method for treating macular degeneration comprising administering the above-mentioned pharmaceutical composition or food composition to a patient with macular degeneration or a subject at risk for macular degeneration, or depending on the formulation of the composition.
  • a method of treatment, prevention or improvement is provided.
  • the subject is a mammal, including a human.
  • the blueberry extract and the blueberry fraction which are the active ingredients of the present invention, have the effect of fundamentally preventing the etiology by inhibiting A2E accumulation in retinal pigment epithelial cells and inhibiting the oxidation of A2E. Therefore, the composition of the present invention can be developed into a variety of products, such as drugs for the treatment or improvement of excellent macular degeneration and health functional food.
  • Figure 1 is a graph showing the test results for the antioxidant activity of the blueberry extract and blueberry fraction for A2E oxidized by ultraviolet A.
  • Figure 2 is a graph showing the test results of the protective effect of the blueberry and its fractions against apoptosis by UVA in ARPE-19 cells accumulated A2E.
  • Figure 3 is a graph showing the test results for the inhibition of the accumulation of A2E and iso A2E by treatment with blueberry extract and blueberry fraction in ARPE-19 cells.
  • the good condition was selected and washed thoroughly. Then, 2000 ml of water was added to 100 g of blueberries, followed by extraction of hot water twice at 16C for 16 hours to obtain a 10% blueberry extract.
  • the blueberry extract obtained as described above was freeze-dried, processed into a powder form, and then stored at -40 ° C. until used to prepare the blueberry fraction and used in the experiments described below.
  • the blueberry extract powder prepared in Example 1 was dissolved in distilled water, followed by connecting two C18 Sep-Pak cartridges, preconditioning with 10 ml ethyl acetate and absolute methanol, and then 0.01 N aqueous HCl. Spilled into cartridge.
  • the whole extract thus obtained was loaded into a cartridge and carbohydrates, acids and other water soluble compounds were removed with 6 ml of 0.01 N aqueous HCl.
  • the layer thus removed was used as a carbohydrate / acid fraction in the following experiment as a comparative example.
  • the solvent of the fraction was concentrated at 40 °C to remove all the residual solvent to prepare a blueberry fraction.
  • the carbohydrate / acid fraction and pigment fraction obtained through this process were dissolved in distilled water, respectively, and the blueberry polyphenol fraction was dissolved in 50% aqueous ethanol, and then stored at -70 ° C. for use in the experiment and analysis described below.
  • ARPE-19 Human adult ARPE cells (ARPE-19; catalog no. CRL-2302) used in the experiment and analysis of the present invention were used from the clergy Medical University Institute of Ophthalmology.
  • the ARPE cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS), 100 U / ml penicillin, and 100 mg / ml streptomycin, and used for the experiment. It was set as.
  • the cells were used for the experiment by inoculating 6-well plates with 5 ⁇ 10 4 cell numbers when used for the experiment.
  • An ultraviolet A lamp (Sankyo Denki, Tokyo, Japan) having a wavelength between 355 nm and 375 nm was used for the oxidation experiment of A2E.
  • the irradiation method of ultraviolet A is to wash the well plate with phosphate buffered saline (PBS) before irradiation, and then open the lid of the well plate with only PBS that the cells remain dry so that the irradiation density of 2.35 ⁇ 0.3 mW / cm2 Ultraviolet A was irradiated with an energy of 3 J / cm 2.
  • PBS phosphate buffered saline
  • the cells were sufficiently dissolved in dimethyl sulfoxide (DMSO), and then the absorbance was measured at 550 nm using an ELISA microplate reader, followed by the percentage of the group (normal control) in which neither A2E nor sample accumulated (normal control). ).
  • DMSO dimethyl sulfoxide
  • the blueberry extract and the blueberry fraction were treated for each concentration, and then cultured for 2 days to accumulate A2E as in the previous experiment.
  • the cells were then lysed using 25 mM HEPES buffer, including protease inhibitors (Complete TM, Roche, Mannheim, Germany) and quantitatively analyzed for A2E accumulation using high performance liquid chromatography (HPLC).
  • HPLC high performance liquid chromatography
  • A2E uses a known concentration of A2E solution for quantification and A2E levels were corrected by measuring the protein by the Bradford method (Bio-Rad Protein Assay, Bio-Rad, Hercules, CA). Samples were used after extracting A2E using CHCl 3 from ARPE-19 cells before HPLC quantitation, and conditions and specific experimental methods of HPLC quantitation were performed as shown below.
  • a reversed C18 column 250 4.6 mm, Cosmosil 5C18, Nacalai Tesque, Osaka, Japan
  • Waters 600E HPLC equipped with the Waters 996 photodiode array detector A.
  • A2E is dissolved by dissolving with acetonitrile in water containing 0.1% trifluoroacetic acid (TFA).
  • TFA trifluoroacetic acid
  • blueberry fraction was 33, 71, 85% lower than the untreated A2E treated cells at 25, 50, 100 ⁇ g / ml concentration of the polyphenol fraction, respectively It can be seen that the high A2E oxidation inhibitory ability.
  • the pigment fraction is 32, 53, 66% lower for the untreated A2E at the same concentration as the polyphenol fraction, respectively.
  • Carbohydrate / acid fractions were found to have no antioxidant activity. Therefore, the blueberry fraction showed the ability to inhibit the A2E oxidation in the order of polyphenol fraction> pigment fraction >> carbohydrate / acid fraction.
  • Non A2E-UV Non A2E-UV
  • A2E-non UV cells not irradiated with ultraviolet A after A2E accumulation
  • the survival rate of cells treated with the blueberry extract 125, 250, 500 ⁇ g / ml concentration is 63, 134, 167% higher than the cell survival rate of the negative control, respectively, Blueberry extract demonstrated concentration dependent protection of ARPE-19 cells.
  • the viability of the cells treated with the polyphenol fraction 12.5, 25, 50, and 100 ⁇ g / ml concentration in the blueberry fraction was 63, 80, 161 based on the cell survival rate of the negative control, respectively.
  • the highest cell protection was shown by 207%, and the cell survival rate of the pigment fraction was 14, 26, 44, 75% higher than that of the negative control at the same concentration as the polyphenol fraction.
  • Blueberry carbohydrate / acid fractions on the other hand, were found to be incapable of protecting. Therefore, the blueberry fraction was found to protect the ARPE-19 cells from oxidation by UVA in the order of polyphenol fraction »pigment fraction> carbohydrate / acid fraction.
  • A2E / isoA2E concentrations accumulated in ARPE-19 cells treated with 50, 100, 250, and 500 ⁇ g / ml concentrations of blueberry extracts were measured. 18, 26, 37, 34% / 8, 11, 18, 13% (A2E / iso A2E) lower than the blueberry extract was found to inhibit the accumulation of A2E / iso A2E concentration dependently.
  • the blueberry fraction has a concentration of A2E / isoA2E accumulated in ARPE-19 cells treated with pigment fractions 25, 50, and 100 ⁇ g / ml, compared with cells not treated with blueberry fraction. It was 36, 45, 51/13, 14, 40% lower, showing the highest inhibition of A2E / iso A2E accumulation, and the concentration of A2E / isoA2E accumulated in cells treated with polyphenol fractions was 19, 20, 40 at the same concentration. / 0.1, 3.3, 15% lower.
  • Carbohydrate / acid fractions on the other hand, were found to be incapable of inhibiting A2E / isoA2E accumulation. Therefore, the blueberry fraction demonstrated the effect of inhibiting A2E and isoA2E accumulation in ARPE-19 cells in the order of pigment fraction> polyphenol fraction »carbohydrate / acid fraction.
  • the blueberry extract of the present invention and the fractions fractionated therefrom protect ARPE-19 cells by inhibiting A2E accumulation in ARPE-19 cells and inhibiting oxidation of A2E by UV light. It was found out.
  • composition of the present invention can fundamentally prevent the pathogenesis of macular degeneration, it is expected that the composition of the present invention can be very useful for improving or alleviating macular degeneration.

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PCT/KR2011/001631 2010-03-10 2011-03-09 들쭉 추출물 또는 들쭉 분획물을 유효 성분으로 함유하는 황반 변성증의 치료, 예방 또는 개선용 조성물 WO2011112002A2 (ko)

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CN201180016617.7A CN102821773B (zh) 2010-03-10 2011-03-09 包含笃斯越橘提取物或笃斯越橘部分作为活性成分的用于治疗、预防或改善黄斑变性的组合物
JP2012556977A JP5860418B2 (ja) 2010-03-10 2011-03-09 クロマメノキ抽出物またはクロマメノキ分画物を有効成分として含む黄斑変性症の治療、予防または改善用組成物

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KR10-2010-0021449 2010-03-10

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KR101484386B1 (ko) * 2013-01-23 2015-01-19 정수영 들쭉 추출물로부터 분리된 페놀류 화합물을 유효성분으로 함유하는 황반 변성증의 치료, 예방 또는 개선용 조성물
KR20170036352A (ko) * 2015-09-24 2017-04-03 동아에스티 주식회사 들쭉 추출물을 유효성분으로 함유하는 황반변성증 치료용 조성물
KR102481709B1 (ko) 2015-12-21 2022-12-27 동아제약 주식회사 들쭉 추출물을 유효성분으로 포함하는 안구건조증 치료용 조성물
KR20210142445A (ko) 2020-05-18 2021-11-25 (주)비지엔케어 울금 추출물을 포함하는 망막질환 예방 및 치료용 조성물

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JP2013522188A (ja) 2013-06-13
KR101330850B1 (ko) 2013-11-18
CN102821773A (zh) 2012-12-12

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