WO2005103089A1 - 魚類由来のコンドロイチン硫酸/デルマタン硫酸ハイブリッド鎖 - Google Patents
魚類由来のコンドロイチン硫酸/デルマタン硫酸ハイブリッド鎖 Download PDFInfo
- Publication number
- WO2005103089A1 WO2005103089A1 PCT/JP2005/008326 JP2005008326W WO2005103089A1 WO 2005103089 A1 WO2005103089 A1 WO 2005103089A1 JP 2005008326 W JP2005008326 W JP 2005008326W WO 2005103089 A1 WO2005103089 A1 WO 2005103089A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- unit
- galnac
- glcua
- 3galnac
- chondroitin sulfate
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0069—Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
Definitions
- the present invention relates to a chondroitin sulfate / dermatan sulfate hybrid chain derived from fish, in particular, from shark skin or chrysalis.
- chondroitin sulfate is composed of disaccharides of D-glucuronic acid and N-acetyl-D-galactosamine and sulfate residues. Chondroitin sulfate is classified into types such as A, C, D, E, and K, depending on the binding position of the sulfate group. Chondroitin sulfate L is composed of L-iduronic acid instead of D-glucuronic acid, and is called dermatan sulfate.
- Chondroitin sulphate and dermatan sulphate contain a large number of overlapping sequences built up of disaccharide units with different patterns of sulphation and show huge structural diversity comparable to heparan sulphate.
- the present inventors have worked to elucidate that various polysulfated chondroitin sulfate and dermatan sulfate chains are involved in neural development.
- the “D” unit a polysulfated disaccharide unit in chondroitin sulfate and dermatan sulfate with various biological functions [GlcUA (2S) — GalNAc (6S)], “iD”
- IdoUA stands for L-iduronic acid
- GlcUA stands for D-Darctic acid
- GalNAc stands for N-acetyl-D-galatatosamine
- 2S, 4S and 6S indicate that the binding site of the sulfate group is C-2. Position
- each neuron develops a characteristic morphology and elongates its axon along a specific pathway to find the correct target cell and form a synapse.
- Many researchers have found that treating developing brain tissue with chondroitinase ABC and decomposing and removing chondroitin sulfate contained in the tissue causes major abnormalities in neuronal morphogenesis and axonal running. I have. These observations suggest that chondroitin sulfate is involved in neurite axon guidance or in morphological differentiation of neurons.
- Such polysulfated chondroitin sulfate and dermatan sulfate are known to bind to many heparin-binding growth factors, and their signaling is involved in morphogenesis of hippocampal neurons. Is suggested.
- Non-Patent Document 1 Hikino, M., Mikami, ⁇ ⁇ , Faissner, A., Vilela-Silva, AC, Pavao, MS, and Sugahara, K. (2003) J. Biol. Chera. 278, 43744-43754 Literature 2 Seno, N. and Meyer, K., Biochim. Biophys. Acta. 78 (1963) 258-264.
- a new chondroitin sulfate / dermatan sulfate hybrid chain is separated and purified from a part of the fish body excluding teleost fish such as shark skin, and this fraction binds to various growth factors and promotes neurite outgrowth. It has been found that it has an anticoagulant effect, and can be expected to have an effect as a composition for treating neurological diseases. Accordingly, the present invention provides the following aspects:
- GlcUA (2S) -GalNAc (4S) B unit
- GlcUA-GalNAc (4S) A unit
- GlcUA-GalNAc (6S) C unit
- GlcUA (2S) -GalNAc (6S) D unit
- at least one unit selected from the group consisting of GlcUA-GalNAc (4S, 6S) E unit
- unsulfated GlcUA-GalNAc (0 unit)
- IdoUA IdoUA.
- chondroitin sulfate / dermatan sulfate hybrid chain according to any one of the above 1 to 3, wherein the degree of sulfation per disaccharide molecule is in the range of 0.70 or more and less than 1.2.
- a glycosaminodalican comprising the chondroitin sulfate / dermatan sulfate hybrid chain according to any of 1 to 5 above;
- a growth factor binding agent comprising the glycosaminodalican according to 6 above as an active ingredient
- a neurite outgrowth promoting agent comprising the glycosaminoglycan according to 6 above as an active ingredient
- An anticoagulant comprising the glycosaminoglycan according to 6 above as an active ingredient
- a pharmaceutical composition comprising the glycosaminodalican according to 6 above as an active ingredient;
- a pharmaceutical composition for preventing or treating a neurodegenerative disease comprising the glycosaminodalican according to 6 above as an active ingredient;
- glycosaminodalican as described in 6 above for producing a growth factor binding agent, a neurite outgrowth promoter, an anticoagulant or a composition for treating a neurodegenerative disease
- GlcUA-GalNAc (4S) (A unit), GlcUA (2S)-GalNAc (4S) (B unit).
- GlcUA-GalNAc (6S) (C unit), GlcUA (2S) -GalNAc (6S) ( D unit), GlcUA-GalNAc (4S, 6S) (E unit), GlcUA (2S); 3l-3GalNAc (4S, 6S) (T unit) and unsulfated GlcUA-GalNAc (0 [ At least one unit selected from the group consisting of IdoUA al-3GalNAc (4S) (iA unit), IdoUA (2S) -GalNAc (4S) (iB unit), and IdoUA ⁇ 1 -3GalNAc (6S) (iCut), IdoUA (2S) -GalNAc (6S) (iD unit), IdoUA- GalNAc (4S, 6S) (iE unit) and IdoUA (2S) al-
- a glycosaminodalican comprising the chondroitin sulfate / dermatan sulfate hybrid chain according to 13 above, comprising at least one unit selected from the group consisting of 3GalNAc (4S, 6S) (iTut); 15.
- a growth factor binding agent comprising as an active ingredient the daricosaminodalican according to 13 or 14 above;
- a neurite outgrowth promoter comprising the glycosaminoglycan according to 13 or 14 above as an active ingredient
- a composition for treating a neurodegenerative disease comprising the glycosaminoglycan according to the above 13 or 14 as an active ingredient;
- a pharmaceutical composition comprising the glycosaminodalican as described in 13 or 14 above as an active ingredient;
- a pharmaceutical composition for preventing or treating a neurodegenerative disease comprising the glycosaminoglycan according to the above 13 or 14 as an active ingredient;
- daricosaminoglycan according to 13 or 14 above for producing a growth factor binding agent, a neurite outgrowth promoting agent, or a composition for treating a neurodegenerative disease.
- FIG. 1 shows the results of anion-exchange HPLC analysis of chondroitinase digests of shark skin-derived chondroitin sulfate / dermatan sulfate hybrid chains (SS-CS / DS) (Native).
- FIG. 2 is a diagram showing the results of gel filtration analysis of digests of isolated SS-CS / DS (Native) with various chondroitinases.
- SS-CS / DS (Native) (1 ⁇ g) with chondroitinase AC-1 (A), chondroitinase B (B), or chondroitinase B followed by chondroitinase AC-1 (C)
- the digest was labeled with 2AB and analyzed on a Superdex Peptide column.
- the elution positions of the standard disaccharide oligosaccharides are indicated by arrows: 1 (CS-10 saccharide); 2 (CS-8 saccharide); 3 (CS-6 saccharide); 4 (CS-4 saccharide); Five
- FIG. 3 shows SS-CS / DS (Native) ( ⁇ ), SS-CS / DS1.0M NaCl elution fraction (1.0M) ( ⁇ ), and SS-CS / DS1.5M for molecular size determination.
- FIG. 4 shows the results of gel filtration column mouth chromatography on Superdex-200 performed on the NaCl-eluting fraction (1.5 M) (fraction).
- FIG. 4 shows the binding of various growth factors to SS-CS / DS (1.0 M, 1.5 M).
- A The results of the responses to SS-CS / DS (1.0M) (open bars) and SS-CS / DS (1.5M) (solid bars) are shown.
- B The results of the response to heparin are shown as a control experiment.
- FIG. 6 is a diagram showing the results of analyzing the binding between SS-CS / DS (1.5 M) and BDNF or GDNF using a BIAcore J system.
- the black bars are SS-CS / DS (1.5M), and the white bars are chondroitin sulfate / dermatan sulfate hybrid chains (HN-
- B and C show sensorgrams (B: BDNF and C: GDNF) obtained with various concentrations of BDNF and GDNF. Arrows indicate the start of each of the association and dissociation phases.
- Fig. 7-1 shows the neurite outgrowth-promoting action of three SS-CS / DS preparations (Native, 1.0M, 1.5M).
- Three samples of SS-CS / DS (Native, 1.0M, 1.5M) (0.67 // g as peronic acid) were coated on a cover glass pre-coated with P-0RN, E16 Primary cultures of hippocampal neurons derived from mouse embryos were cultured thereon for 24 hours in the minimum essential medium for idal.
- the average length of the longest neurites is 100 randomly selected cells cultured on three SS-CS / DS preparations (Native, 1.0M, 1.5M). Neurons were weighed.
- Chondroitin sulfate chain (CS-E) derived from squid cartilage was designated as positive control (P ⁇ 0RN), and the coat with P-0RN alone was designated as negative control / (P-0RN) ( ⁇ : p ⁇ 0.001, ⁇ : p ⁇ 0 . 005).
- B is the result of similarly examining SS-CS / DS (Native) digested with various chondroitinases (ns: no significant difference, ⁇ : p ⁇ 0. 001, *: p ⁇ 0. 05) ).
- the number of neurites induced by each fraction not digested with the enzyme is graphed (n.s .: no significant difference, ⁇ : p ⁇ 0.001).
- the value obtained from two separate experiments is expressed as the average soil SE.
- Figure 7-2 is a photograph showing the neurite outgrowth-promoting effect of three SS-CS / DS preparations (Native, 1.0M, 1.5M).
- D shows the morphology of neurons cultured on SS-DS
- E shows the morphology of neurons cultured on a cover glass coated only with P-0RN.
- FIG. 8 shows the results of measuring the anti-factor Ila activity of three samples of SS-CS / DS (Native, 1.0M, 1.5).
- Heparin (Hata) and DS (Garden) derived from pig skin are positive control It is a rule.
- SS-CS / DS (Native) ( ⁇ ), SS-CS / DS (1.OM) ( ⁇ ), and SS-CS / DS (1.5M) (X) were tested.
- Figure 9-1 shows a sensorgram of immobilized HN-CS / DS depending on the growth factor concentration.
- the growth factors tested are FGF-2 (A) and FGF-10 (B).
- FIG. 9-2 is a diagram showing a sensorgram of immobilized HN-CS / DS depending on the growth factor concentration.
- the growth factors tested are FGF-16 (C) and FGF-18 (D).
- FIG. 9-3 is a diagram showing a sensorgram of immobilized HN-CS / DS depending on the growth factor concentration.
- the growth factors tested are position (E) and PTN (F).
- FIG. 9-4 is a diagram showing a sensorgram of immobilized HN-CS / DS depending on the growth factor concentration.
- the growth factors tested are HB-EGF (G) and VEGF (H).
- FIG. 10 is a diagram showing sensorgrams superimposed when HN-CS / DS is immobilized on a sensor chip and BDNF and GDNF are allowed to flow on the sensor chip at various concentrations.
- FIG. 11-1 is a photograph showing the neurite outgrowth-promoting effect of the HN-CS / DS digest.
- A is a neuron cultured on a cover glass coated with HN-CS / DS, and B is a cell cultured only on the P-0RN coat on the cover glass.
- FIG. 11-2 is a diagram showing the neurite outgrowth-promoting effect of the HN-CS / DS digest.
- C indicates the length of the neurites, and D indicates the number of neurites.
- the present invention relates to a chondroitin sulfate / dermatan sulfate hybrid chain and glycosaminodalican containing the same. Further, the present invention relates to a chondroitin sulfate / dermatan sulfate hybrid chain containing a disaccharide of GlcUA (2S) -GalNAc (4S), and a glycosaminoglycan containing the same.
- Such glycosaminoglycan of the Haiburitsu de chain 60% or more, preferably 70% or more, more preferably 80% or more, further preferred properly 90% or more, particularly preferably 95% or more (all w / w ), Most preferably, no other sugar chain component is detected, and glycosaminoglycans substantially consisting only of the above-mentioned hybrid chains.
- the chondroitin sulfate / dermatan sulfate hybrid chain and daricosaminodalican containing the same in the present invention are covalently bonded to the bran chain.
- the chondroitin sulfate / dermatan sulfate hybrid chain refers to a polysaccharide chain having a disaccharide unit structure, including both a disaccharide unit possessed by chondroitin sulfate and a disaccharide unit possessed by dermatan sulfate.
- the type and content of the disaccharide unit containing the chondroitin sulfate / dermatan sulfate hybrid chain are not limited. For example, the following structure has been isolated from chondroitin sulfate.
- Such a chondroitin sulfate / dermatan sulfate hybrid chain can be separated and purified from fish bodies other than teleost fish.
- fish other than teleost fish include cartilaginous fish and fish belonging to the conch.
- the fish body is not limited, but can be appropriately selected depending on the type of fish, the content of the chondroitin sulfate / dermatan sulfate hybrid chain, and the ease of separation / purification.
- the notochord of shark skin ⁇ ⁇ can be used. From the sample, acetone extraction and protease treatment.
- the daricosaminodarican fraction obtained by extraction with triclo-mouth acetate was further subjected to CPC precipitation, hyaluronidase digestion, nitrite treatment, desalting, and chondroitin sulfate / dermatan sulfate.
- a hybrid chain fraction is obtained.
- the chondroitin sulfate / dermatan sulfate hybrid chain fraction with an anion exchange resin the chondroitin sulfate / dermatan sulfate hybrid chain adsorbed on the resin is removed.
- fractions of displacement has a substantially equal molecular weight, the average molecular weight, for example, shark skin-derived chondroitin sulfate / dermatan sulfate Haipuriddo chain cases, about 65 ⁇ 7 5 kDa, preferably about 68 ⁇ 7 2 kDa, and most preferably about 70 kDa, if the j Asian swamp eel notochord derived chondroitin sulfate / dermatan sulfate Haipuriddo chain is about 18 kDa.
- shark skin-derived chondroitin sulfate / dermatan sulfate Haipuriddo chain cases about 65 ⁇ 7 5 kDa, preferably about 68 ⁇ 7 2 kDa, and most preferably about 70 kDa, if the j Asian swamp eel notochord derived chondroitin sulfate / dermatan sulfate Haipuriddo
- the degree of sulfation per disaccharide molecule of the chondroitin sulfate / dermatan sulfate hybrid chain derived from shark skin in the present invention is less than 1.2, that is, preferably about 0.70 or more and less than 1.20, more preferably about 0.80 or more and less than about 1.20, particularly preferably in the range of about 0.87 to 1.17.
- Each step of separating and purifying chondroitin sulfate / dermatan sulfate hybrid chains from fish other than teleost fish such as shark skin and eel chords may be performed by methods known to those skilled in the art. As an example, embodiments of a series of steps are described below in the Examples section, but are not limited to the described method.
- chondroitin sulfate / dermatan sulfate hybrid chain may be referred to as CS / DS.
- those derived from shark skin may be prefixed with SS (shark skin), and those derived from dung eel chord may be prefixed with HN (hagfish notochord).
- the fraction obtained from the above-mentioned hyaluronidase digestion, nitrous acid treatment, and desalting from a shark skin sample is subjected to SS-CS / DS (Native) and further to an anion exchange resin to obtain about 1.0 M N
- the fraction eluted with .aCl and the fraction eluted with about 1.5M NaCl are called SS-CS / DS (1.0M) and SS-CS / DS (1.5M), respectively.
- each fraction of these chondroitin sulfate / dermatan sulfate hybrid chains derived from shark skin was performed by digestion with chondroitinase ABC, B or AC-1.
- the major component of each SS-CS / DS fraction was GlcUA j3 1-3GalNAc (4S) (A unit) or]: doUA o _3GalNAc (4S) (iA unit) and GlcUA ⁇ l-3GalNAc (6S) (Cunit) or IdoUAall-3GalNAc (6S) (iCunit).
- chondroitin sulfate and dermatan sulfate in these fractions consisted of very diverse disaccharide molecules, and in particular, disaccharide molecules having the structure of GlcUA (2S) -GalNAc (4S) were detected.
- Such a chondroitin sulfate / dermatan sulfate hybrid chain containing a disaccharide molecule having the structure of GlcUA (2S) -GalNAc (4S) was found for the first time in the present invention, and chondroitin sulfate / dermatan sulfate derived from shark skin. It is considered to be a property peculiar to the hybrid town.
- the chondroitin sulfate / dermatan sulfate hybrid chain derived from shark skin always contains GlcUA (2S)-GalNAc (4S) (B unit), and further contains GlcUA-
- GalNAc (4S) A unit
- GlcUA- GalNAc (6S) C unit
- One unit ie, one, two, three, four or four units
- one IdoUA 1 3GalNAc (4S) iA unit
- IdoUA-GalNAc (4S, 6S) iD unit
- at least one unit ie one, two, three, four or five units selected from the group consisting of IdoUA-GalNAc (4S, 6S) (iE unit). Including. Since it is a chondroitin sulfate / dermatan sulfate hybrid chain, GlcUA (2S)-GalNAc (4S) (B unit) and GlcUA-GalNAc (4S)
- At least one member of the group of chondroitin sulfate units consisting of GlcUA-GalNAc (0 [unit]) and IdoUA al-3GalNAc (4S) (iA unit), IdoUA (2S) -GalNAc (4S)
- IB unit IdoUA al-3GalNAc (6S) (iC unit), IdoUA (2S) -GalNAc (6S) (iD unit) and at least one of the dermatan sulfate units consisting of IdoUA-GalNAc (4S, 6S) (iE unit).
- the chondroitin sulfate / dermatan sulfate hybrid chain derived from the eel chord of the green eel GlcUA ⁇ l-3GalNAc (4S) (A unit), GlcUA ⁇ 1-3GalNAc (6S) (C unit), GlcUA j8 3GalNAc (4S, 6S) (E unit), GlcUA (2S) ⁇ At least one unit selected from the group consisting of l-3GalNAc (4S, 6S) (T unit) and unsulfated GlcUA-GalNAc (0 unit) (ie, one, two, three units) , 4 or 5 units), and IdoUA ⁇ 1-3GalNAc (4S).
- IA unit IdoUA11GalNAc (6S) (iC unit), IdoUA11-3GalNAc (4S) , 6S) (iE unit) and IdoUA (2S) 1-3GalNAc (4S, 6S) (iT unit) at least one unit selected from the group consisting of forces (ie one, two, three or 4 units).
- chondroitin sulfate / dermatan sulfate hybrid chains derived from fish other than teleost fish including sharks and blue eels are GlcUA-GalNAc (4S) (A unit).
- the chondroitin sulfate / dermatan sulfate hybrid chain of the present invention can be used for various growth factors (fibroblast growth factor (FGF) -2, FGF-10, FGF-18, heparin-binding epithelial cell growth factor (HB-EGF)). , Mitodocaine (MK), pleiotropin pin (PTN), brain-derived neurotrophic factor (BDNF) and glial cell-derived neurotrophic factor (GDNF) with high affinity.
- FGF fibroblast growth factor
- FGF-10 fibroblast growth factor
- FGF-18 heparin-binding epithelial cell growth factor
- HB-EGF heparin-binding epithelial cell growth factor
- MK Mitodocaine
- PDN pleiotropin pin
- BDNF brain-derived neurotrophic factor
- GDNF glial cell-derived neurotrophic factor
- the chondroitin sulfate / dermatan sulfate hybrid chain of the present invention exhibits a neurite outgrowth promoting action and an anticoagulant action. .
- chondroitin sulfate / dermatan sulfate hybrid chains derived from fish other than teleost fish, such as those derived from shark skin and scallop chordae are useful as therapeutic or preventive agents for neurodegenerative diseases and promotion of neural system development. Furthermore, it is useful as an anticoagulant.
- the present invention relates to a glycosaminoglycan comprising a chondroitin sulfate / dermatan sulfate hybrid chain derived from fish other than teleost fish, such as a chondroitin sulfate / dermatan sulfate hybrid chain contained in the above-mentioned fraction derived from shark skin and hagfish chord.
- growth factors FGF-2, FGF-10, FGF-18, HB-EGF, MK, PTN, BDNF and GDNF binding agents, neurite outgrowth promoters, and anticoagulants.
- daricosaminodalican containing a chondroitin sulfate / dermatan sulfate hybrid chain is completely or partially purified from shark skin, and the obtained fraction is, if necessary, added to a suitable carrier and / or solvent. It is good to mix with.
- the daricosaminodalican can be used as a pharmaceutical composition.
- the glycosaminoglycan can be used as a pharmaceutical composition for preventing or treating neurodegenerative diseases. Since a neurodegenerative disease involves degeneration and loss of neurites of nerve cells, the glycosaminodalican can prevent this or promote the growth of new neurites, so that the progression or symptom of the disease is caused by the disease. Can be reduced.
- ⁇ 20 / _f g / ml preferably :! ⁇ 10 g / ml, more preferably :! At a concentration of ⁇ 5 / ig / ml, It is preferred that they act.
- glycosaminodalican containing the above-mentioned chondroitin sulfate / dermatan sulfate hybrid chain or a fraction containing the same is applied as a neurite outgrowth promoter
- a glycoprotein containing a chondroitin sulfate / dermatan sulfate hybrid chain as described above is obtained from shark skin.
- the fraction obtained by fully or partially purifying the saminoglycan is administered in such a way that it can contact the nerve cells of the target individual in need of promoting neurite outgrowth. be able to.
- a person skilled in the art can select an appropriate administration method, but it is preferable to administer it locally so that the elongation can be delivered to a nerve cell or a periphery of the desired nerve cell. It is preferable to administer such that the concentration at the time of delivery is about 1 to 10 ⁇ g / ml, more preferably about 1 to 5 / z g / ml, and still more preferably about 2 ⁇ g / ml.
- the same applies to fish other than teleost fish such as glycosaminoglycans containing chondroitin sulfate / dermatan sulfate hybrid chains derived from the eel chord.
- the above-mentioned dalicosaminodalican or a fraction containing it is applied as a growth factor binding agent, an appropriate amount (depending on the target growth factor) of the above glycosaminodalican or The fraction containing it is mixed with a growth factor and brought into contact.
- glycosaminoglycans Dali cans 0. l / g or more, preferably mixed with the blood at a concentration of 1 ⁇ 10 ⁇ ⁇ / ⁇ 1, by contacting, anticoagulant activity is exhibited.
- the subject of these applications may be any mammal.
- the skin of the blue shark Prionace glauca (85 g dry weight) was defatted three times by acetone extraction and air-dried completely. After suspending in water, the suspension was kept in boiling water for 30 minutes to inactivate the protease.
- borate-NaOH buffer and calcium chloride to a final concentration of 0.1 and 10 respectively, and for 24 hours at 60 ° C protease (actinase: 2% by weight of the sample) Digested. And after 24 hours and 48 hours, allowed to newly Akuchinaze after (1 wt 0/0 of the sample) was added digestion, centrifuged added so that 50% of trichloroacetic port acetate to 5% final concentration, precipitates proteins Then, each suspension was resuspended in 5% trichloroacetic acid and centrifuged again to recover each supernatant.
- glycosaminoglycan-containing precipitate is solubilized with 0.02 M Na 2 SO 4 , and 10% (w / v) cetylpyridinium chloride (CPC) /0.02 M Na 2 SC ⁇ is added.
- CPC cetylpyridinium chloride
- the obtained flocculent sediment was redissolved in a 100: 15 (v / v) 2M NaCl / ethanol solution, and 3 volumes of 99.5% ethanol was added to precipitate again.
- the above steps were repeated three times and finally precipitated in water and dried.
- a similar CPC precipitation step was performed while maintaining the critical electrolyte concentration at 0.5 M NaCl to remove hyaluronic acid and concentrate glycosaminodalican.
- the precipitate obtained after hyaluronidase digestion were treated with 40 minutes at room temperature to stand at is nitrous acid obtained by mixing 0. 5 mmole nitrite Bariumu with the same amount of 0. 5 mmol e sulfate. A freshly prepared aliquot of nitrite was added again and allowed to stand for another 40 minutes.
- the desalted glycosaminodalican was passed through a Sep-Pak C18 cartridge column and eluted with water, followed by 100% methanol.
- the free peptide-free chondroitin sulfate / dermatan sulfate hybrid chain eluted here was called SS-CS / DS (Native).
- This preparation contains a small amount of a peptide covalently bonded to a sugar chain (about 0.6 to 1.2% by weight).
- SS-CS / DS (Native) is passed through an anion exchanger cartridge and stepped with 300 mM phosphate buffer containing 0.15, 0.5, 1.0, 1.5 or 2.0 M NaCl. After elution, the mixture was desalted with 50 mM pyridine acetate buffer (pH 5.0; eluent) using PD-10 force column.
- the fractions eluted with the 1.0 and 1.5 M buffers containing NaCl contain 20% and 80% chondroitin sulfate / dermatan sulfate hybrid chains, respectively, and the SS-CS / DS ( 1.0M) and SS-CS / DS (1.5M).
- Table 1 shows the purification yields.
- table 1 SS-CS / DS yield
- Disaccharide composition analysis was performed by analyzing digests of chondroitinase ABC, B or AC-1.
- the disaccharide labeled with 2AB is 16 mM Diluted by 1 and analyzed by anion exchange HPLC (PA-03 silica column using 16mM and 530mM Na ⁇ PO ⁇ solvents).
- the 3/3 samples (Native, 1.0M, 1.5M) were digested with lOmlU chondroitinase ABC (total volume of 10 il), and 1/3 each was digested with lOmlU chondro 4- or chondro 6-.
- the digest was subjected to sulfatase digestion, and the digest was labeled with 2AB and analyzed by HPLC as described above.
- Table 2 shows the results of the digestion and analysis of three SS-CS / DS samples (Native, l.OM, 1.5M).
- SS-CS / DS (Native), SS-CS / DS (l.OM) and SS-CS / DS (1.5M) all have AHexUAal-3GalNAc (ADi-OS), ⁇ at various ratios HexUA Hi 1—3GalNAc (6S)
- the main constituents of the three SS-CS / DS standards are ⁇ Di-4S and ⁇ Di-6S, SS-CS / DS (Native), SS-CS / DS ( l.OM) and the ratio of ADi-4S and ⁇ -6S to the total disaccharide in SS-CS / DS (1.5M) were 48.9% and 21.3 ° /, respectively. , 43.5%, 21.3%, 55.0% and 16.6% (Table 2).
- the ratio of ⁇ Di-0S is SS-CS / DS (1.0M) force 23.8%, compared with SS-CS / DS (Native) (10.5%) and SS-CS / DS (1.5M) (5.8%) And it was expensive.
- SS- CS / DS (1.5M) is disulfated disaccharide (ADi - diS D, ADi - diS D Oyopi ADi - diS c) ratio of is 22.6%
- SS-CS / DS (Native) has a wide variety of different sulfation degrees.
- chondroitin sulfate and dermatan sulfate derived from marine organisms including shark cartilage, squid cartilage, sea squirt, sea squirt, sea urchin and sea eel, which have been isolated so far, are ⁇ E '' unit, ⁇ iE '' unit, and ⁇ D Unit, iD unit or iBJ
- iA unit or IdoUA (2S) -GalNAc (4S) (iB unit), but rather as GlcUA-GalNAc (4S) (A unit) or GlcUA (2S) -GalNAc (4S) (Bunit) Suggest to do. Also, the majority of the iA units or iB units do not form clusters and are therefore considered to be resistant to chondroitinase B digestion.
- Chondroitinase B did not release A Di-6S. Since the chondroitinase was detected to some extent after AC-1 digestion, the ability of all 6-0 sulfated units to exist as GlcUA-GalNAc (6S) (C unit) or IdoUA-GalNAc (6S) ( (iC unit). These may be resistant to the enzyme or may be contained in sequences that are resistant to the enzyme.
- the SS-CS / DS sample was hardly detected by the chondroitinase AC-1 digestion of three SS-CS / DS samples (Native, 1.0M, 1.5M), and the SS-CS / DS sample Non-sulfated and monosulfated disaccharides, including ADi-0S, ADi-6S and ADi-4S, were detected in only about 10% of the total amount of disaccharides in them (Table 2). This result suggests that at least a part of the 4-0 sulfated unit and the 6-0 sulfated unit are present as “A” and “CJ units”.
- Decasaccharide or larger oligosaccharides were less in chondroitinase AC-I digest (FIG. 2A) than in chondroitinase B digest (FIG. 2B). This was in favor of a somewhat higher ratio of GlcUA than IdoUA.
- the disaccharide obtained by digestion of either chondroitinase probably consists of a unit containing GlcUA (10 ° /.) And a unit containing IdoUA (2%). Indicates the presence of adjacent sequences.
- fractions corresponding to the two disaccharides chondroitinase AC- 1 digests, respectively A Di-diS E as a major component, A Di as a minor component - there is diS B To do so.
- disulfated disaccharide chondroitinase B digest, A Di- diS B as main components, A Di as a small amount component - indicates that there is diS E.
- the “B” unit containing GlcUA (GlcUA (2S) -GalNAc (4S)) was detected for the first time. Although the values for these units shown in Table 2 are very low, they are only cuttable units. Therefore, it is highly probable that other B units are frequently included. This B unit may be involved in the function of SS-CS / DS.
- the molecular size of the chondroitin sulfate / dermatan sulfate hybrid chain was measured by gel filtration on a Superdex 200 column (10 ⁇ 300 mm). Three samples of SS-CS / DS (Native, 1. OM, 1.5 M) (40 ⁇ g) were loaded on the column, and eluted with 0.2 M ammonium acetate at a flow rate of 0.3 ml / min. The eluted fraction for 3 minutes was collected in one fraction, evaporated to dryness, and dissolved in 100 1 water. Measurements were as previously reported (Chandrasekhar, S., et al (1987) Anal Biochem. 161, 103-108).
- This molecular weight is as follows: chrysalis of chrysalis (18 kDa), pig skin (l 9 kDa), eel skin
- any SS-CS / DS preparation (1.0 ⁇ , 1.5M) Also showed a high interaction response with all growth factors except FGF-1, but the response was dependent on the growth factor.
- the degree of coupling also differs depending on the SS-CS / DS standard (1.0M, 1.5M), and SS-CS / DS (1.5M) is higher than SS-CS / DS (1.0M) Responsiveness was induced ( Figure 4).
- SS-CS / DS In terms of the number of growth factors that bind to glycosaminodalican chains of the same length, SS-CS / DS (1.5M) has several times higher binding ability than CS-H, even when the binding ability is expressed. (Table 3, right column). Despite several times lower than Hep, the degree of sulfation of SS-CS / DS (1.5 M)
- Hippocampus were isolated from el 6 mouse embryos - Yulong dispersed on the cover glass at a density of 10, 000 cells / mm 2 a, 37 ° under C0 2 5% by Eagle's minimum essential medium (:, 24 hours The cells were then fixed and immunostained using monoclonal antibodies to neurofilament (NF) and microtubule-associated protein (MAP2).
- NF neurofilament
- MAP2 microtubule-associated protein
- Immunostained cells on each coverslip were counted under a microscope. The length of the longest neurites and the number of neurites in randomly selected cells were measured using image analysis software (MacSCOPE; Mitani Corporation, Fukui, Japan).
- the neurite outgrowth-promoting activity is indicated by the length of the longest neurite outstretched.
- the neurite outgrowth promotion was observed in the three samples of SS-CS / DS (Native, 1.0 ⁇ , 1.5 ⁇ ). Also exceeded CS--(Fig. 7-1A).
- the neurite outgrowth promoting activity was higher in the order of SS-CS / DS (Native), SS-CS / DS (1.5M), and then SS-CS / DS (1.0M).
- Anti-factor Ila activity is 1 ⁇ 10 // ⁇ / ⁇ 1 SS-CS / DS containing 3 standards (Native, 1.0M, 1.5M) 50mM Tris-HC1 buffer containing 227mM NaCl 100 ⁇ 1 ( ⁇ 8.3) and normal human plasma 60 ⁇ 1 and human thrombin (1.2 ⁇ units / ml) 40 zl were measured by incubating at 25 ° C for 1 minute (Sakai, S et al. (2003) Carbohydr. Res. 338, 263-269). The araidolytic activity of thrombin was determined by measuring the change in absorbance at 405 nm for 100 seconds after adding 100 1 of 1.9 mol / ml chromozym TH. The change in absorbance was proportional to the residual thrombin activity. Heparin and dermatan sulfate from pig skin were used as positive control.
- SS-CS / DS Three samples of SS-CS / DS (Native, 1.0M, 1.5M) showed the same inhibition of thrombin activity as the positive control. SS-CS / DS (Native) showed slightly higher anti-factor Ila activity than the other two SS-CS / DS (1.0 M 1.5 M) (FIG. 8).
- Notochord of hagfish Eptatretus burgeri was treated with acetone, and the suspension was digested with pronase. The protein was precipitated and centrifuged by adding 20% trichloroacetic acid, and the supernatant was collected. Two times the amount of ethanol containing 2.5% calcium acetate was added to precipitate daricosaminodalican. Darikosami Nogurikan mixture through Dowex 1-C1- column and eluted stepwise with NaCl solution was removed by dividing the heparan sulfate for 2 ⁇ 0M fraction to follow the method of Shively and Cornad. After nitrous acid treatment, neutralized with Na 2 C0 3 in 0.
- the fraction eluted with 2.0 M NaCl contained a chondroitin sulfate / dermatan sulfate hybrid chain (CS / DS chain).
- the CS / DS chain thus isolated from the notochord of the hagfish Eptatretus burgeri is referred to herein as HN-CS / DS.
- Chondroitin sulfate / dermata is obtained by gel filtration on Superdex 200 column (10 X 300 mm). The molecular size of the CS / DS chain was measured. The HN- CS / DS and loaded into column and eluted with acetic acid Anmoniumu of 0. 2M at a flow rate of 0. 3 ml / min. The eluted fraction for 3 minutes was collected in one fraction, evaporated to dryness, and dissolved in 100 1 water. Measurement already reported
- HN-CS / DS When the molecules of HN-CS / DS were examined by gel filtration column chromatography, the average molecular weight of HN-CS / DS was 18 kDa, which was close to that of dermatan sulfate (19 kDa) derived from pig skin and dermatan sulfate (14 kDa) derived from eel skin. was something.
- Disaccharide composition analysis was performed by analyzing digests of chondroitinase ABC, B or AC-1.
- HN-CS / DS was digested with either lOmlU chondroitinase ABC, 5 mlU chondroitinase AC-1, or lmlU chondroitinase B, and the digests were reported (Kinoshita , A., Sugahara, K (1999) Anal. Biochera. 269, 367-378), labeled with 2-aminobenzamide (2AB) (excess 2AB is water: cloth form 1: 1 (v / v) Removed with the mixture).
- 2-aminobenzamide (2AB 2-aminobenzamide
- HN-CS / DS was digested with chondroitinase ABC, AC-1 or B.
- Orchid-CS / DS is composed mainly of A HexUA and 1-3GalNAc (4S, 6S) ( ⁇ Di-diS E ), ⁇ HexUA was 1-3GalNAc (4S) (ADi-4S). Also, AHexUA ⁇ 1-3GalNAc (6S) Di-6S), ⁇ HexUA al-3GalNAc ( ⁇ Di—OS), ⁇ HexUA (2S) al-3GalNAc (4S, (ADi-triS) (Table 6) Table 6
- HN-CS / DS The interaction analysis between HN-CS / DS and various growth factors (FGF-2, FGF-10, FGF-16, FGF-18, PTN, MK, HB-EGF, VEGF) was performed using the BIAcore system (BIAcore, Uppsala, Sweden). Power rice tick parameters k a, k d Oyopi were calculated using the BIA evaluation 3 ⁇ 1 software.
- Fig. 9-1 shows the FGF-2 (A), FGF-10 ( ⁇ ) (Fig. 9-1), FGF-16 (C), and FGF-18 (Fig. 9-4) of HN-CS / DS.
- D (Fig. 9-2), ⁇ ( ⁇ ), PTN (F) (Fig. 9-3), HB-EGF (G), VEGF (H) (Fig. 9-4). All growth factors examined showed a typical growth factor binding pattern.
- ⁇ -Analysis of the interaction between CS / DS, BDNF and GDNF was performed using the BIAcore J system (BIAcore, Uppsala, Sweden). Biotin-labeled micro-CS / DS was immobilized on a sensor chip coated with streptavidin, and BDNF and GDNF were allowed to flow at various concentrations over the sensor chip (high flow rate (601 / min)). . The binding phase was 2 minutes and the dissociation phase was 3 minutes. The force parameters k a , k d and K d were calculated using BIA evaluation 3.1 software (Table 8).
- HN-CS / DS is fixed on the sensor chip, and BDNF and GDNF are mixed at various concentrations.
- Figure 10 shows the sensorgram when it was flown over the sensor chip.
- HN-CS / DS The affinity of HN-CS / DS for GDNF was about 10 times higher than that of heparan sulfate (HS) and GDNF, and the affinity for BDNF was significantly higher at 360 times. These high binding affinities are due to chondroitin sulfate, dermatan sulfate or
- NF eurofilament
- MAP2 microtubule-associated protein
- the immunostained cells of each coverslip were counted under a microscope.
- the length of the longest neurites and the number of neurites in randomly selected cells were measured using Image Analysis Software (Mac SCOPE; Mitani Corp., Tokyo, Japan).
- the neurite outgrowth-promoting activity was indicated by the length of the longest neurite outgrowth, indicating that neurite outgrowth was promoted as compared to control (Fig. 11-12C).
- the present invention provides a novel chondroitin sulfate / dermatan sulfate hybrid chain and a fraction containing the same, which bind to various growth factors and exhibit a neurite outgrowth promoting action and an anticoagulant action.
- a composition for neurological treatment is provided.
- the chondroitin sulfate / dermatan sulfate hybrid chain of the present invention is derived from fishes other than teleost fish such as shark skin and hagfish, and useful uses of fish have been found by the present invention. All publications cited herein are incorporated by reference in their entirety. It will be readily apparent to those skilled in the art that various modifications and alterations of the present invention are possible without departing from the technical concept and the scope of the invention described in the appended claims. The present invention is intended to cover such modifications and variations.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Neurology (AREA)
- Biomedical Technology (AREA)
- Neurosurgery (AREA)
- Materials Engineering (AREA)
- Epidemiology (AREA)
- Dermatology (AREA)
- Biochemistry (AREA)
- Polymers & Plastics (AREA)
- Hospice & Palliative Care (AREA)
- Psychiatry (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP05736922A EP1746108A1 (en) | 2004-04-27 | 2005-04-25 | Fish-origin chondroitin sulfate/dermatan sulfate hybrid chain |
US11/587,789 US20070232564A1 (en) | 2004-04-27 | 2005-04-25 | Fish-Origin Chondroitin Sulfate/Dermatan Sulfate Hybrid Chain |
JP2006512664A JPWO2005103089A1 (ja) | 2004-04-27 | 2005-04-25 | 魚類由来のコンドロイチン硫酸/デルマタン硫酸ハイブリッド鎖 |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US56551104P | 2004-04-27 | 2004-04-27 | |
US60/565,511 | 2004-04-27 | ||
JP2004308584 | 2004-10-22 | ||
JP2004-308584 | 2004-10-22 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2005103089A1 true WO2005103089A1 (ja) | 2005-11-03 |
Family
ID=46045414
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2005/008326 WO2005103089A1 (ja) | 2004-04-27 | 2005-04-25 | 魚類由来のコンドロイチン硫酸/デルマタン硫酸ハイブリッド鎖 |
Country Status (4)
Country | Link |
---|---|
US (1) | US20070232564A1 (ja) |
EP (1) | EP1746108A1 (ja) |
JP (1) | JPWO2005103089A1 (ja) |
WO (1) | WO2005103089A1 (ja) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2007016099A (ja) * | 2005-07-06 | 2007-01-25 | Japan Science & Technology Agency | プレイオトロフィン結合性および非結合性のコンドロイチン硫酸/デルマタン硫酸オリゴ糖 |
JP2007224286A (ja) * | 2006-01-25 | 2007-09-06 | Tottori Univ | 生体組織からの多糖類の取得方法 |
JP2007291322A (ja) * | 2006-03-31 | 2007-11-08 | Maruha Corp | 新規多糖およびオリゴ糖 |
US8481711B2 (en) | 2010-07-06 | 2013-07-09 | Hidenori Kamanishi | Neurite outgrowth agent |
JP2020173228A (ja) * | 2019-04-12 | 2020-10-22 | 株式会社島津製作所 | グリコサミノグリカンの分析方法 |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101633701B (zh) * | 2008-07-23 | 2011-10-05 | 上海景峰制药有限公司 | 一种透明质酸的纯化方法 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0930979A (ja) * | 1995-07-24 | 1997-02-04 | Seikagaku Kogyo Co Ltd | 神経疾患の治療剤 |
-
2005
- 2005-04-25 US US11/587,789 patent/US20070232564A1/en not_active Abandoned
- 2005-04-25 JP JP2006512664A patent/JPWO2005103089A1/ja active Pending
- 2005-04-25 WO PCT/JP2005/008326 patent/WO2005103089A1/ja active Application Filing
- 2005-04-25 EP EP05736922A patent/EP1746108A1/en not_active Withdrawn
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0930979A (ja) * | 1995-07-24 | 1997-02-04 | Seikagaku Kogyo Co Ltd | 神経疾患の治療剤 |
Non-Patent Citations (8)
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2007016099A (ja) * | 2005-07-06 | 2007-01-25 | Japan Science & Technology Agency | プレイオトロフィン結合性および非結合性のコンドロイチン硫酸/デルマタン硫酸オリゴ糖 |
JP2007224286A (ja) * | 2006-01-25 | 2007-09-06 | Tottori Univ | 生体組織からの多糖類の取得方法 |
JP2007291322A (ja) * | 2006-03-31 | 2007-11-08 | Maruha Corp | 新規多糖およびオリゴ糖 |
US8481711B2 (en) | 2010-07-06 | 2013-07-09 | Hidenori Kamanishi | Neurite outgrowth agent |
JP2020173228A (ja) * | 2019-04-12 | 2020-10-22 | 株式会社島津製作所 | グリコサミノグリカンの分析方法 |
JP7185232B2 (ja) | 2019-04-12 | 2022-12-07 | 株式会社島津製作所 | グリコサミノグリカンの分析方法 |
Also Published As
Publication number | Publication date |
---|---|
JPWO2005103089A1 (ja) | 2008-03-13 |
EP1746108A1 (en) | 2007-01-24 |
US20070232564A1 (en) | 2007-10-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Shriver et al. | Heparin and heparan sulfate: analyzing structure and microheterogeneity | |
Shi et al. | Glycosaminoglycan-protein interactions and their roles in human disease | |
CN103739743B (zh) | 糖胺聚糖的分离和鉴定 | |
Meneghetti et al. | Heparan sulfate and heparin interactions with proteins | |
Gandhi et al. | The structure of glycosaminoglycans and their interactions with proteins | |
Coombe et al. | Heparan sulfate-protein interactions: therapeutic potential through structure-function insights | |
Westergren-Thorsson et al. | Increased deposition of glycosaminoglycans and altered structure of heparan sulfate in idiopathic pulmonary fibrosis | |
Lyon et al. | Interaction of hepatocyte growth factor with heparan sulfate. Elucidation of the major heparan sulfate structural determinants | |
Chen et al. | The structure property and endothelial protective activity of fucoidan from Laminaria japonica | |
Bolten et al. | Heparin: role in protein purification and substitution with animal-component free material | |
Zhang et al. | Modification, characterization and structure–anticoagulant activity relationships of persimmon polysaccharides | |
Pan et al. | Glycosaminoglycans from fish swim bladder: isolation, structural characterization and bioactive potential | |
Kunze et al. | Sulfated hyaluronan derivatives reduce the proliferation rate of primary rat calvarial osteoblasts | |
CN104470530B (zh) | 来自prasinococcale的多糖 | |
WO2006083328A2 (en) | Biologically active surfaces and methods of their use | |
WO2005103089A1 (ja) | 魚類由来のコンドロイチン硫酸/デルマタン硫酸ハイブリッド鎖 | |
Ikeda et al. | Synthesis and biological activities of a library of glycosaminoglycans mimetic oligosaccharides | |
Coltrini et al. | Biochemical bases of the interaction of human basic fibroblast growth factor with glycosaminoglycans: new insights from trypsin digestion studies | |
Volpi | Dermatan sulfate: Recent structural and activity data | |
JP6741277B2 (ja) | 硫酸化ヘパリン由来オリゴ糖及びその調製法と施用 | |
Yates et al. | Highly diverse heparan sulfate analogue libraries: providing access to expanded areas of sequence space for bioactivity screening | |
Ramachandra et al. | Brittlestars contain highly sulfated chondroitin sulfates/dermatan sulfates that promote fibroblast growth factor 2-induced cell signaling | |
Kaneda et al. | Structural characteristics of heparin-like domain required for interaction of midkine with embryonic neurons | |
JP2004043645A (ja) | 低分子化糖の製造方法 | |
Rapraeger | Heparan sulfate-growth factor interactions |
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 BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KM KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SM SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU MC NL PL PT RO SE SI SK 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 | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2006512664 Country of ref document: JP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 11587789 Country of ref document: US Ref document number: 2007232564 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWW | Wipo information: withdrawn in national office |
Country of ref document: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2005736922 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 2005736922 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 11587789 Country of ref document: US |