+

WO2002076487A1 - Agents anticancereux contenant des peptides antigenotoxiques et immunostimulants produits a partir d'hydrolysate de cocon de vers a soie - Google Patents

Agents anticancereux contenant des peptides antigenotoxiques et immunostimulants produits a partir d'hydrolysate de cocon de vers a soie Download PDF

Info

Publication number
WO2002076487A1
WO2002076487A1 PCT/KR2002/000418 KR0200418W WO02076487A1 WO 2002076487 A1 WO2002076487 A1 WO 2002076487A1 KR 0200418 W KR0200418 W KR 0200418W WO 02076487 A1 WO02076487 A1 WO 02076487A1
Authority
WO
WIPO (PCT)
Prior art keywords
hydrolysate
fibroin
peptides
silk thread
anticancer
Prior art date
Application number
PCT/KR2002/000418
Other languages
English (en)
Inventor
Yuk-Hyun Joo
Chang-Kee Hyun
Sung-Hee Lee
Deock-Hyoung Cho
Geum-Ju Park
Original Assignee
Aminogen Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Aminogen Co., Ltd. filed Critical Aminogen Co., Ltd.
Publication of WO2002076487A1 publication Critical patent/WO2002076487A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/01—Hydrolysed proteins; Derivatives thereof
    • A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1767—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/308—Foods, ingredients or supplements having a functional effect on health having an effect on cancer prevention

Definitions

  • the present invention relates to an anticancer agent comprising peptides of silkworm cocoons as an active ingredient, and more particularly to an anticancer agent comprising, as an active ingredient, peptides obtained by hydrolyzing fibroin extracted from a silk thread or waste silk thread of silkworm cocoons.
  • Silkworms have been raised traditionally in Korea to produce raw silk. With emergence of artificial silk or synthetic fibers, however, competitiveness of silk threads produced from silkworm cocoons was weakened, and also the traditional sericultural industry has been reduced. Recently, such a sericultural industry has pursued to development of hypoglycemic agents utilizing silkworms, production of vegetable worms, health beverages and cosmetics utilizing silk powder, and health foods made from mulberry leaves.
  • Cocoon threads produced from silkworms are composed of fibroin, a fibrous protein, and sericin, a glue protein, which coats the fibroin part.
  • the protein of cocoon threads has an amino acid composition as follows: 45 % glycine, 30 % alanine, 12 % serine, and 5 % tyrosine. That is, main amino acids account for more than 90 % in the composition of silk.
  • Japan with the aim of providing edibility of these proteins, research into how such proteins can be digested into low molecular weight peptides or amino acids by hydrolysis thereof using acids, alkalis, or enzymes, and digested and absorbed into the body in an efficient manner, is underway.
  • 96-0015242 entitled "Therapeutic agent containing silk fibroin for insulin-independent diabetes” discloses a novel use of an aqueous solution of a silk fibroin peptide as a hypoglycemic agent.
  • physiological activities of hydrolysates of the silk protein known so far include promotion of alcohol metabolism, hypoglycemic effect, decrease of level of blood cholesterol, and anti-dementia effect.
  • these activities are not yet systemically studied.
  • few studies on peptides having an anticancer effect have achieved success, despite increasing demand for development of anticancer agents.
  • the present inventors have conducted research to develop an anticancer agent capable of inhibiting DNA damage caused by carcinogens and improving immune activity.
  • the inventors found that hydrolysates of a silk protein, fibroin, extracted from silkworm cocoons have effects of inhibiting DNA damage caused by a foreign carcinogen, and improvement of immune activity by enhancing a macrophage activity, thereby inhibiting proliferation of tumor cells. Accordingly, it is an object of the present invention to provide an anticancer agent comprising, as an active ingredient, anticancer peptides having effects of inhibition of DNA damage caused by carcinogens and improvement of immune activity.
  • an anticancer agent comprising, as an active ingredient, peptides obtained by hydrolyzing fibroin extracted from a silk thread or waste silk thread of silkworm cocoons, using an acid or a protease.
  • a processed food prepared by adding anticancer peptides in powder or other edible formulations, the peptides being obtained by hydrolyzing fibroin extracted from a silk thread or waste silk thread of silkworm cocoons, using an acid or a protease.
  • an anticancer agent comprising a fibroin hydrolysate as an active ingredient according to the invention
  • sericin is removed by treating cocoons with sodium carbonate or sodium bicarbonate.
  • sodium carbonate was used herein at a concentration of 1 to 10 % to remove sericin.
  • fibroin was treated with an acid or protease, preparing a hydrolysate.
  • the acid may be oxalic acid or HC1, and HC1 is more preferable.
  • the concentration of HC1 is preferably 1 to 5 N. More particularly, after treating with HC1, fibroin is heated at 90 to 100 ° C for 4 to 8 hrs, followed by neutralization with sodium hydroxide, adjusting to pH 7.0 to 7.4, thus preparing a hydrolysate.
  • various substances may be used. Especially, an active carbon was used herein.
  • the amount of active carbon is preferably 3 to 10 % relative to a total amount of the solution.
  • the protease may be bacteria- derived proteases and bacteria-derived proteases for use in industry.
  • the proteases used herein are trypsin, pepsin, Alcalase, and Neutrase.
  • each concentration of the enzymes is preferably 0.1 to 5 %.
  • an active peptide fraction was isolated from the hydrolysate.
  • the peptide fraction may be isolated using any common methods known in the art.
  • the peptide fraction was isolated using chromatography, and especially, gel filtration chromatography on Sephadex G-25. It is preferable that a step of obtaining a peptide fraction using Sephadex G-15 from the peptide fraction isolated using the above chromatography is further conducted. In such a way, a peptide fraction with molecular weights between 500 and 1000, exhibiting an increased activity in terms of inhibition of DNA damage, compared to a crude hydrolysate, could be obtained.
  • the hydrolysate isolated from fibroin inhibits DNA damage by a carcinogen.
  • Various carcinogens may be used to induce DNA damage.
  • the carcinogen used herein is a direct acting carcinogen, MNNG (N-methyl-N'-nitro-N-nitrosoguanidine).
  • MNNG N-methyl-N'-nitro-N-nitrosoguanidine
  • Inhibition effects of DNA damage by hydrolysate according to the present invention may be measured by performing Ames test and SOS chromotest.
  • the test used herein is Comet test.
  • the two former tests mentioned above employ Salmonella and E. coli. For this reason, they may produce inaccurate results due to different biological characteristics between microorganisms and animal cells, upon measurement of inhibition of DNA damage with respect to the animal cells.
  • the Comet test used herein it has an advantage in that physiologically active substances inhibiting DNA damage can be screened more accurately.
  • the Comet test is a method for characterizing DNA damage and repair thereof in various mammalian cells. Using this test, it is possible to detect DNA damage which can't be detected by a conventional method. Thus, this method is applicable to various fields including screening of various substances involved in carcinogenesis, genetic toxicology associated with DNA damage and repair thereof, monitoring of environmental pollution, and determination of a cancer prevention effect by Lactobacillus .
  • effects of the fibroin hydrolysate for improving immune activity were measured.
  • a common method known in the art may be used for measuring improvement effect of immune activity.
  • the method used herein is to measure reactive nitrogen species, for example, nitric oxide, produced by macrophages upon treatment of the hydrolysate, the reactive nitrogen species being representative chemical substances involved in immune responses.
  • the increase in production of the reactive nitrogen species directly reflects the increase of TNF (Tumor Necrosis Factor) production. Accordingly, detection of increased production of the reactive nitrogen species indicates an effect of improved immune activity of the hydrolysate.
  • Macrophages responsible for a first line of immune surveillance in the body are known to play a significant role in a defense system against tumors.
  • activated macrophages selectively recognize and eliminate tumor cells through a direct contact with the cells.
  • Such activated macrophages secret various cytokines, nitric oxide and hydrogen peroxide.
  • secretion of nitric oxide by macrophages, induced by the hydrolysate was measured.
  • amounts of nitric oxide were increased by peptide components of the hydrolysate, resulting in improving immune activity, which is attributable to the hydrolysate, as shown in Fig. 2.
  • hydrolysate of the invention exhibits the effects as anticancer substances.
  • the invention is also directed to a processed food prepared by adding anticancer peptides in powder or other edible formulations, the peptides being obtained by hydrolyzing fibroin extracted from a silk thread or waste silk thread of silkworm cocoons, using an acid or a protease.
  • the fibroin hydrolysate according to the invention exhibits an anticancer effect, it is possible to produce functional foods containing the hydrolysate. Ingestion of such foods can inhibit DNA damage caused by a foreign carcinogen, and improve immune activity. If the fibroin hydrolysate containing peptides is added to foods in an edible formulation, the formulation is not limited. But a powder form of the hydrolysate is preferable for addition to processed foods.
  • Fig. la is a result of gel filtration chromatography using Sephadex G-25 with respect to a fibroin hydrolysate
  • Fig. lb is a result of gel filtration chromatography using Sephadex G-15 with respect to a peptide fraction corresponding to a third peak of Fig. la;
  • Fig. 2 is a graph showing a change in amounts of nitric oxide produced upon treatment of a fibroin hydrolysate to intraperitoneal macrophages of mouse.
  • Selected cocoons were treated with 10 % sodium carbonate (Na 2 CO 3 ) and heated for 1 hr. The resulting solution was filtered, removing solubilized sericin, thereby obtaining a silk thread consisting of fibroin.
  • One part of the cocoon consisting of fibroin obtained as in Example 1-1 was added with 80 parts by weight of 2 N HC1, and heated at 100 ° C for 48 firs, thus hydrolyzing fibroin.
  • the hydrolysate thus prepared was dark brown.
  • the neutralized hydrolysate was then added with active carbon in an amount of 6 % relative to a total amount of the solution.
  • the solution was stirred for 60 min to remove diverse non-dissolved substances and abnormal odor generated in the process.
  • the solution was then filtered, obtaining a clear liquid hydrolysate.
  • the hydrolysate solution was finally dialyzed with distilled water for 1 day, using a dialysis membrane, thereby removing salts.
  • Example 1-3 Preparation of enzymatic hydrolysate The silk thread consisting of fibroin obtained as in Example 1-1 was digested into a variety of sizes of peptides by using protease.
  • the silk thread consisting of fibroin was added with a 30 % CaCl 2 solution and heated at 90 ° C for 30 min to solubilize it.
  • the solubilized fibroin was then dialyzed with distilled water for 3 days. After dialysis, the resulting solution was treated with trypsin (Sigma) at 37 ° C (pH7.5), pepsin (Sigma) at 37 ° C (pH 2.0),
  • the hydrolysate thus prepared was centrifuged at 13,000 rpm for 15 min. The supernatant was collected and subjected to ultrafiltration using a centrifugal ultrafiltration kit (Vivaspin 20, Satorius, Germany). The filtrate was assayed for an anticancer effect. Meanwhile, with respect to the filtrates prepared thus, peptide quantification was performed by the Lowry method. Based on differences between the measurements of the peptide and the concentrations of proteins measured before hydrolysis, peptide yields, that is, degree of hydrolysis (DH), were calculated. Concentration and reaction time of enzymes were determined by finding the maximal hydrolysis conditions where DH is not further increased with increased reaction time.
  • DH degree of hydrolysis
  • the cell line used herein was a normal mouse embryo 3T3 cell line (ATCC CCL No. 163), provided from the Korean Collection for Type Cultures (KCTC).
  • KCTC Korean Collection for Type Cultures
  • DMEM Gibco, USA
  • fetal calf serum Hyclon, USA
  • penicillin G Sigma
  • streptomycin sulfate Sigma, USA
  • the 3T3 cell line was maintained on a 10 cm round culture dish (Falcon, USA) containing the DMEM medium at 37 ° C, and 5 % CO 2 in a humidified incubator (Labline instruments, USA). The cells were subcultured every 3 days.
  • the direct acting carcinogen, N-methyl-N'-nitro-N-nitrosoguanidine was diluted in HBSS (10 mM, pH 7.4) to a concentration of 100 g/ml.
  • HBSS HBSS
  • Each of the hydrolysates obtained as in Example 1 was also dissolved in HBSS (10 mM, pH 7.4) to concentrations of 10 mg/ml, 5 mg/ml, 2 mg/ml, and 1 mg/ml, respectively.
  • 10 ⁇ i of MNNG dissolved in HBSS in the above way was added, giving a final concentration of 1 g/ml MNNG.
  • the samples were pre-incubated for 30 min. As a negative control, HBSS only was used.
  • HBSS containing MNNG at a final concentration of 1 zg/ml was used as a positive control.
  • each sample was added to the 3T3 cells, which had been cultured to a density of 1 x 10 5 cells/plate on 3 cm diameter round culture dishes (Falcon, USA) at 37 ° C, and 5 % CO 2 in the humidified incubator, followed by incubation for 30 min.
  • NMA Normal Melting Point Agarose, Sigma, USA
  • PBS Normal Melting Point
  • ERIE Scientific a fully frosted slide glass
  • the slide glass was again smoothly applied with 75 ⁇ l of the NMA solution, using a cover glass (24 x 50 mm, Superior, Germany), and the agarose layer was hardened by placing the slide glass on ice in a flat stainless steel tray.
  • the pre-coated agarose gel slide was transferred to a humidified slide box and stored at 4 ° C .
  • the 3T3 cells were cultured for 72 hrs, while being subcultured by placing 3 x 10 4 cells/plate on a 3 cm round culture dish. After 4 days, the cells were washed twice with Ca 2+ , Mg 2+ -free PBS. The cells were then treated with a mixture of MNNG and the hydrolysate, the hydrolysate being at a concentration of 10 mg/ml, 5 mg/ml, 2 mg/ml, or 1 mg/ml, prepared as in Example 2-2. The cells treated with HBSS containing MNNG served as a positive control, while the cells treated with HBSS only served as a negative control.
  • the cells thus treated were incubated at 37 ° C for 30 min at a speed of 25 rpm in a shaking incubator. After incubation, the cells were immediately washed three times with PBS. With a 100 ⁇ l of proteinase K (Sigma, USA) solution, the cells were completely detached from the tissue culture dish. The cells were resuspended in a 1 ml culture medium and transferred to an Eppendorf tube. The cell suspension was centrifuged at 1,000 rpm for 5 min, and the supernatant was discarded. The cell pellet was resuspended in a 300 ⁇ l of medium. From this, 30 ⁇ i was again centrifuged, obtaining a cell pellet. 3-3: Comet test
  • the cell pellet obtained as in Example 3-2 which had been treated with a mixture of MNNG and varying concentrations of the hydrolysate, HBSS containing MNNG, or HBSS only, were suspended in 75 ⁇ l of a 0.75 % LMA (Low Melting Point Agarose) solution which had been maintained at 45 °C in a water bath.
  • LMA Low Melting Point Agarose
  • the cell suspension mixed with LMA was transferred onto the coated slide prepared as in Example 3-1, using a pipet.
  • the slide was covered with a coverslip to ensure that the cell suspension was smoothly spread, and placed in a flat stainless steel tray filled with ice, to harden for 10 min.
  • Electrophoresis was then conducted at 25V/300mA for 20 min. After electrophoresis, each slide was washed with a neutralization buffer. The slide was stained with 80 ⁇ l of YOYO-1 (Molecular Probes, USA), a fluorescent dye, at a concentration of 10 g/ml to examine the degree of DNA damage in the 3T3 cells treated with a mixture of MNNG and varying concentrations of the hydrolysate, HBSS containing MNNG, or HBSS only, as in Example 3-2. The stained slide was observed at a magnification of 250X, using a fluorescent microscope (CSB-FEI, China) with a 200 W mercury lamp (Osram, Germany).
  • Tail moment hydrolysate treatment (mean ⁇ SD) (mean ⁇ SD)
  • the sample 1 refers to cells with a certain degree of DNA damage where the hydrolysate and MNNG were pre-incubated, allowing a reaction therebetween, and then the mixture was added to the 3T3 cells.
  • the sample 2 refers to cells with a certain degree of DNA damage where the hydrolysate and MNNG were separately added to the 3T3 cells, without pre-incubation thereof.
  • Each of the samples 3 and 4 refers to cells with a certain degree of DNA damage where the hydrolysate was first pre-incubated with the 3T3 cells for 30 min, allowing a reaction between the peptides and cells to occur.
  • the hydrolysate added to the cells was washed three times with PBS, and then the cells were incubated with MNNG for 30 min, that is, without the hydrolysate.
  • the cells were further incubated with MNNG for 30 min, with the hydrolysate.
  • no effect of inhibition of DNA damage was seen in the sample 2.
  • the samples 3 and 4 exhibited reduced DNA damage. Consequently, it can be inferred that the mechanism by which the hydrolysate inhibits DNA damage is by reacting with the carcinogen and cells, thereby functioning to inhibit DNA damage.
  • each of the hydrolysates obtained as in Example 1 was subjected to gel filtration chromatography, and fractionated according to molecular weights of peptides. With respect to respective peaks for the hydrolysate, the degrees of inhibition of DNA damage were measured.
  • Gel filtration chromatography on Sephadex G-25 showed three peaks, having molecular weights between 1,000 and 5,000, as shown in Fig. la. The peptides corresponding to respective peaks were separated and each peptide fraction was lyophilized.
  • the degrees of inhibition of DNA damage versus a carcinogen were measured.
  • the fraction of a third peak whose activity had been confirmed, was refractionated using Sephadex G-15.
  • the chromatography showed again three peaks. Among those fractions, the peptides corresponding to a third peak were subjected to the
  • mice Culture of macrophage BALB/c mice were injected intraperitoneally with 1 ml of a 1 % solution of thioglycollate. After 4 days, the mice were sacrificed by cervical dislocation. 10 ml of RPMI 1640 was injected to the abdominal cavity, and intraperitoneal cells were collected after flushing three times, followed by centrifuging. The intraperitoneal cells were suspended in RPMI 1640 containing 10 % FBS, and 200 ⁇ l each was plated on a 96 well plate. The cell number per well was 2 x 10 5 . The plate was incubated at 37 ° C under 5 % CO 2 .
  • the cells were washed with phosphate buffered saline (pH 7.4) to remove non-adherent cells from the plate.
  • the adherent macrophages were subject to continued incubation.
  • the filtered sterile hydrolysate sample was added at a concentration of 0.01 to 10 mg/ml, and interferon- ⁇ was added after 16 hrs, thereby activating macrophages.
  • a portion of the culture was collected, and the amount of nitric oxide produced during incubation was measured.
  • the cell culture obtained by treating with 1 ⁇ gl l of lipopolysaccharide served as a positive control.
  • Amounts of nitric oxide, one of reactive nitrogen species produced from macrophages, were measured by quantifying amounts of nitrite accumulated in the culture of the activated macrophages.
  • the present invention provides an anticancer agent comprising peptides obtained from the fibroin hydrolysate which was extracted from silk threads or waste silk threads of silkworm cocoons.
  • the peptides are capable of inhibiting DNA damage caused by foreign carcinogens and improving immune activity.
  • Those peptides of the invention may be applied in the future to produce medications and functional foods exhibiting anticancer effects, thereby contributing to development of medicines for prevention and treatment of cancer.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Nutrition Science (AREA)
  • Zoology (AREA)
  • Epidemiology (AREA)
  • Polymers & Plastics (AREA)
  • Food Science & Technology (AREA)
  • Immunology (AREA)
  • Mycology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne des agents anticancéreux comprenant des peptides produits à partir d'un cocon de vers à soie utilisé en tant que composant efficace. Plus particulièrement, l'invention concerne des agents anticancéreux contenant des peptides produits par hydrolyse d'une fibroïne obtenue par extraction d'un fil de soie ou d'un fil de soie de schappe d'un cocon de vers à soie. Selon cette invention, les peptides anticancéreux pouvant inhiber l'endommagement de l'ADN par des carcinogènes étrangers et améliorer l'activité immunitaire, peuvent être produits à partir d'hydrolysate de fibroïne extraite du fil de soie ou du fil de soie de schappe d'un cocon de vers à soie. Les peptides anticancéreux décrits dans cette invention peuvent être appliqués à des médicaments et à des aliments fonctionnels ayant une activité anticancéreuse, de sorte qu'ils peuvent servir au développement de médicaments destinés à la prévention ou au traitement du cancer.
PCT/KR2002/000418 2001-03-21 2002-03-11 Agents anticancereux contenant des peptides antigenotoxiques et immunostimulants produits a partir d'hydrolysate de cocon de vers a soie WO2002076487A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020010014706A KR20020074746A (ko) 2001-03-21 2001-03-21 누에고치 단백질로부터 제조된 항유전독성 및 면역활성증강 효과를 갖는 항암제
KR2001/14706 2001-03-21

Publications (1)

Publication Number Publication Date
WO2002076487A1 true WO2002076487A1 (fr) 2002-10-03

Family

ID=19707207

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2002/000418 WO2002076487A1 (fr) 2001-03-21 2002-03-11 Agents anticancereux contenant des peptides antigenotoxiques et immunostimulants produits a partir d'hydrolysate de cocon de vers a soie

Country Status (2)

Country Link
KR (1) KR20020074746A (fr)
WO (1) WO2002076487A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006014033A1 (fr) 2004-07-31 2006-02-09 Brainguard Co., Ltd. Peptide de soie ameliorant les effets neuroprotecteurs et neurofonctionnels et methode de preparation
WO2007048985A2 (fr) * 2005-10-28 2007-05-03 Engelhard Lyon Substance pour restaurer une co-expression et une interaction normales entre les proteines lox et nrage
CN103613652A (zh) * 2013-11-15 2014-03-05 苏州大学 一种丝素蛋白的提纯方法
EP2748177A1 (fr) * 2011-08-26 2014-07-02 Agricultural Research Development Agency (Public Organization) Compositions oligopeptidiques biologiquement actives à base de soie et leur procédé de fabrication
CN105331661A (zh) * 2015-11-24 2016-02-17 浙江汇能生物股份有限公司 一种水酶法制备丝胶蛋白多肽的方法
CN107099571A (zh) * 2017-04-28 2017-08-29 安徽生物肽产业研究院有限公司 一种仿生酶解制备蚕蛹小肽的生产方法

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101036796B1 (ko) * 2008-04-17 2011-05-25 화인코주식회사 미세 캡슐을 포함하는 에멀젼 및 미세 캡슐 제조 방법
KR101359502B1 (ko) * 2012-06-18 2014-02-10 대한민국 누에를 이용한 골다공증 예방 및 개선용 식품 조성물 및 이를 함유하는 건강보조식품
KR101966892B1 (ko) * 2017-03-06 2019-04-09 농업회사법인 에스에스바이오팜 주식회사 효소분해에 의한 실크피브로인의 분해방법.
KR102590057B1 (ko) 2023-03-14 2023-10-17 (주)씨앤비오 누에추출 단백질을 이용한 식품조성물

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH10298199A (ja) * 1997-04-22 1998-11-10 Puroza Tec:Kk 新規ペプチド及びそれを含有するアンギオテンシンｉ変換酵素阻害剤
KR100207935B1 (ko) * 1996-05-09 1999-07-15 조재연 견피브로인을 함유하는 인슐린 비의존형 당뇨병치료제
CN1230548A (zh) * 1998-12-22 1999-10-06 金明官 丝素肽的催化循环制备法

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4755325B2 (ja) * 1997-11-04 2011-08-24 一丸ファルコス株式会社 絹蛋白質加水分解物由来の生理活性作用組成物
KR100286388B1 (ko) * 1998-03-14 2001-05-02 대한민국 효소분해에 의한 실크분말 펩타이드의 제조방법
KR19980081941A (ko) * 1998-08-06 1998-11-25 조병남 누에고치 등을 이용한 기능성 실크아미노산, 펩티드 소재의 제조방법 중 효소 분해법에 의한 제조방법
KR100383798B1 (ko) * 2000-03-02 2003-05-12 이영우 실크 펩타이드를 함유하는 기능성 음료의 제조방법
KR20010025612A (ko) * 2001-01-11 2001-04-06 조석형 저분자 펩타이드의 제조방법
KR101183290B1 (ko) * 2010-02-19 2012-09-14 주식회사화신 버링장치 및 그 제어방법
KR101194147B1 (ko) * 2010-04-19 2012-10-23 연세대학교 산학협력단 상변화 물질을 함유한 나노 캡슐의 제조방법

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100207935B1 (ko) * 1996-05-09 1999-07-15 조재연 견피브로인을 함유하는 인슐린 비의존형 당뇨병치료제
JPH10298199A (ja) * 1997-04-22 1998-11-10 Puroza Tec:Kk 新規ペプチド及びそれを含有するアンギオテンシンｉ変換酵素阻害剤
CN1230548A (zh) * 1998-12-22 1999-10-06 金明官 丝素肽的催化循环制备法

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHEN K. ET AL.: "Production of soluble fibroin powder by hydrolysis with hydrochloric acid and physical properties", NIPPON SANSHIGAKU ZASSHI, vol. 60, no. 5, 1991, pages 358 - 362 *
GOTOH K. ET AL.: "Sulfated fibroin, a novel sulfated peptide derived from silk, inhibits human immunodeficiency virus replication in vitro", BIOSCI. BIOTECHNOL. BIOCHEM., vol. 64, no. 8, 2000, pages 1664 - 1670 *
HIGUCHI A. ET AL.: "Enhanced CEA production associated with aspirin in a culture of CW2 cells on some polymeric films", CYTOTECHNOLOGY, vol. 31, no. 3, 1999, pages 233 - 242, XP019236661, DOI: doi:10.1023/A:1008030730814 *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1773864A1 (fr) * 2004-07-31 2007-04-18 Biogrand Co., Ltd. Peptide de soie ameliorant les effets neuroprotecteurs et neurofonctionnels et methode de preparation
EP1773864A4 (fr) * 2004-07-31 2009-02-18 Biogrand Co Ltd Peptide de soie ameliorant les effets neuroprotecteurs et neurofonctionnels et methode de preparation
US20110105402A1 (en) * 2004-07-31 2011-05-05 Brainguard Co., Ltd. Silk Peptide For Improving Neuroprotective And Neurofunctional Effects And A Method Of Its Prepartion
AU2004322006B2 (en) * 2004-07-31 2012-05-24 Brainguard Co., Ltd. Silk peptide improving neuroprotective and neurofunctional effects and a method of its preparation
WO2006014033A1 (fr) 2004-07-31 2006-02-09 Brainguard Co., Ltd. Peptide de soie ameliorant les effets neuroprotecteurs et neurofonctionnels et methode de preparation
US10010574B2 (en) * 2004-07-31 2018-07-03 Brainguard Co., Ltd. Silk peptide for improving neuroprotective and neurofunctional effects and a method of its preparation
US9308161B2 (en) 2005-10-28 2016-04-12 Basf Beauty Care Solutions France S.A.S. Substance for restoring normal co-expression and interaction between the LOX and NRAGE proteins
WO2007048985A2 (fr) * 2005-10-28 2007-05-03 Engelhard Lyon Substance pour restaurer une co-expression et une interaction normales entre les proteines lox et nrage
WO2007048985A3 (fr) * 2005-10-28 2008-04-10 Engelhard Lyon Substance pour restaurer une co-expression et une interaction normales entre les proteines lox et nrage
EP2345405A1 (fr) * 2005-10-28 2011-07-20 BASF Beauty Care Solutions France SAS Substance pour restaurer une co-expression et une interaction normales entre les protéines LOX et NRAGE
EP2748177A1 (fr) * 2011-08-26 2014-07-02 Agricultural Research Development Agency (Public Organization) Compositions oligopeptidiques biologiquement actives à base de soie et leur procédé de fabrication
EP2748177A4 (fr) * 2011-08-26 2015-04-08 Agricultural Res Dev Agency Public Organization Compositions oligopeptidiques biologiquement actives à base de soie et leur procédé de fabrication
CN103613652A (zh) * 2013-11-15 2014-03-05 苏州大学 一种丝素蛋白的提纯方法
CN103613652B (zh) * 2013-11-15 2015-12-30 苏州大学 一种丝素蛋白的提纯方法
CN105331661A (zh) * 2015-11-24 2016-02-17 浙江汇能生物股份有限公司 一种水酶法制备丝胶蛋白多肽的方法
CN107099571A (zh) * 2017-04-28 2017-08-29 安徽生物肽产业研究院有限公司 一种仿生酶解制备蚕蛹小肽的生产方法

Also Published As

Publication number Publication date
KR20020074746A (ko) 2002-10-04

Similar Documents

Publication Publication Date Title
AU2007343749B2 (en) Milk protein hydrolyzates with reduced immunogenic potential
MATAR et al. Immunomodulating effects of milks fermented by Lactobacillus helveticus and its non-proteolytic variant
JP5689222B2 (ja) コラーゲンペプチド組成物及びこれを含有する飲食品
TWI659746B (zh) 包含膠原蛋白胜肽、彈力蛋白胜肽以及蛋白聚醣之組成物
Li et al. Purification and identification of novel peptides with inhibitory effect against angiotensin I-converting enzyme and optimization of process conditions in milk fermented with the yeast Kluyveromyces marxianus
Hao et al. Proteolysis and ACE-inhibitory peptide profile of Cheddar cheese: Effect of digestion treatment and different probiotics
Xing et al. Gelling behavior of bio-tofu coagulated by microbial transglutaminase combined with lactic acid bacteria
Ghassem et al. Kinetic characterization of Channa striatus muscle sarcoplasmic and myofibrillar protein hydrolysates
WO2002076487A1 (fr) Agents anticancereux contenant des peptides antigenotoxiques et immunostimulants produits a partir d'hydrolysate de cocon de vers a soie
KR100699324B1 (ko) 어린단백질 가수분해물의 제조방법
Yuan et al. Effects of peptide–calcium complexes from sunflower seeds and peanuts on enhancing bone mineral density
Ahmed et al. In-vitro self-assembly and antioxidant properties of collagen type I from Lutjanus erythropterus, and Pampus argenteus skin
CA2765992A1 (fr) Hydrolysat de proteines de poissons pour son utilisation dans l'inhibition de la prise de poids et/ou la perte de poids
Diao et al. Anti-inflammatory effects of mung bean protein hydrolysate on the lipopolysaccharide-induced RAW264. 7 macrophages
CN108935912A (zh) 一种具有dpp-ⅳ抑制和抗疲劳功能的鱼肉蛋白肽及其制备方法
Zhang et al. Silver carp (Hypophthalmichthys molitrix) by-product hydrolysates: A new nitrogen source for Bifidobacterium animalis ssp. lactis BB-12
Hua et al. Hydrolyzing condition and immunocompetence of sheep bone protein enzymatic lysates
KR102644928B1 (ko) 갈색거저리 유충 단백질을 이용한 근육 질환의 예방 또는 개선용 두부의 제조방법
RU2665589C2 (ru) Способ получения гидролизата рыбного коллагена
KR102755539B1 (ko) 갈색거저리 유충 단백질을 이용한 근육 질환의 예방 또는 개선용 두부
CN114073754A (zh) 大鲵骨粉提取物、制备方法及其应用
CA3164511A1 (fr) Hydrolysats de collagene de dinde et procedes de preparation
KR20020023866A (ko) 누에고치 소재 단백가수분해법에 의한 실크 아미노산과 실크 펩타이드의 제조방법 및 이의 항암, 항유전독성 효과
KR100370883B1 (ko) 대장균에 대해 항균활성을 갖는 한국산 흑염소 락토페린 유래 가수분해물 및 펩타이드
CN112126669B (zh) 免疫调节用的泥丁肽及其制备方法

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SD SE SG SI SK SL TJ TM TN TR TT TZ UA UG US UZ VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载