WO2006098332A1 - 硬組織形成促進剤 - Google Patents
硬組織形成促進剤 Download PDFInfo
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- WO2006098332A1 WO2006098332A1 PCT/JP2006/305052 JP2006305052W WO2006098332A1 WO 2006098332 A1 WO2006098332 A1 WO 2006098332A1 JP 2006305052 W JP2006305052 W JP 2006305052W WO 2006098332 A1 WO2006098332 A1 WO 2006098332A1
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-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- 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
Definitions
- the present invention relates to a hard tissue formation promoter containing glycosaminodarlican or a salt thereof.
- ALP alkaline phosphatase
- a MEM amost, J, essential 3 ⁇ 4 ⁇ ground (minimum essential medium alpha medium)
- BMP-2 Bone morphogenetic protein-2
- BSP bone sialoprotein
- FBS fetal bovine serum
- GAG glycosaminoglycan
- GlcN Gnorecosamine (glucosamine)
- HexA Hexuronic acid
- RT reverse transcription PCR
- 2DSH 2— O—Desulfated Hep (2-0-desulfated heparin) (Hydoxyl group at HexA residue in Hep is desulfated)
- 6DSH 6— 0—Desulfated Hep (6-0-desulfated heparin) (Hydoxyl group at position 6 of GlcN residue in Hep is desulfated)
- NDSH N—Desulfated Hep (N-desulfated heparin) (position 2 of GlcN residue in Hep) Of the amino group is desulfurized)
- Patent Document 1 contains, as an active ingredient, a GAG having a sulfate group and having a repeating structure of a disaccharide consisting of a HexA residue and a GlcN residue as an active ingredient, and the GAG has a 2-position hydroxyl group.
- an intercellular communication enhancer characterized in that it is sulfated and the HexA residue or 2-amino group is sulfaminated to contain a GlcN residue.
- Patent Document 2 contains, as an active ingredient, a GAG having a sulfate group and having a repeating structure of a disaccharide consisting of a HexA residue and a GlcN residue as a basic skeleton.
- an intercellular communication inhibitor characterized in that a sulfhydryl group is sulfated to contain a ⁇ Glc residue.
- Patent Document 3 discloses a certain kind of desulfated ⁇ Hep, which is a basic fibroblast growth factor activity promoter containing this as an active ingredient, and basic fibroblast growth Disclosed is a composition in which the activity of a basic fibroblast growth factor containing the factor is enhanced for cell proliferation.
- Patent Document 4 discloses a hyaluronic acid fraction having an osteoinductive activity having a molecular weight of 20 K Daltonka et al. And 40 K Dalton.
- Patent Document 1 Japanese Patent Application Laid-Open No. 2003-113090
- Patent Document 2 Japanese Patent Laid-Open No. 2003-119146
- Patent Document 3 International Publication No. 96Z01278 Pamphlet
- Patent Document 4 Japanese Patent No. 3333205
- a disaccharide repeating structure consisting of a HexA residue and a GlcN residue is retained as a basic skeleton, and the hydroxyl group at the 2-position of the HexA residue in the basic skeleton, the 6-position of the GlcN residue.
- GAG or its salt itself directly promotes the formation of hard tissue, and the sulfate group is retained in any one or less of the hydroxyl group of GlcN residue and the amino group at the 2-position of GlcN residue. It has the effect of promoting cell differentiation and enhancing the ALP activity of cells! / Neither disclosed nor suggested! / ⁇ .
- An object of the present invention is to provide an agent that directly acts on a cell to cause the promotion of hard tissue formation, the promotion of cell differentiation, and the enhancement of the ALP activity of the cell.
- the present inventor directly acts on a certain type of GAG or its salt cell to promote hard tissue formation, cell differentiation and cell AL.
- the present invention has a sulfate group and has the following properties (1) and (2):
- a hard tissue forming agent (hereinafter referred to as “the present invention formation accelerator”) containing AG or a salt thereof as an active ingredient
- the basic skeleton is a disaccharide repeat structure consisting of HexA residues and GlcN residues.
- the HexA residue is preferably a GlcA residue or an IdoA residue.
- the hard tissue is preferably bone.
- the present invention also relates to a cell differentiation promoting agent (hereinafter referred to as "the present invention") comprising GAG having a sulfate group and having the following characteristics (1) and (2) or a salt thereof as an active ingredient. It is called “differentiation promoter”.
- the basic skeleton is a disaccharide repeat structure consisting of HexA residues and GlcN residues.
- the HexA residue is preferably a GlcA residue or an IdoA residue.
- GAGs that retain sulfate groups and have the above-described characteristics are specifically Hep and Hep. Hold the sulfate group at the 2nd hydroxyl group of the xA residue! /, NA! /, Hold the sulfate group at the 6th hydroxyl group of the GlcN residue in Hep! /, NA! /, 1 or 2 or more selected from the group forces consisting of those having no sulfate group in the amino group at the 2-position of the GlcN residue in Hep.
- the cells are preferably bone marrow-derived mesenchymal cells or osteoblasts.
- the present invention provides a cellular ALP activity enhancer (hereinafter referred to as “this") containing GAG having a sulfate group and the following properties (1) and (2) or a salt thereof as an active ingredient.
- this a cellular ALP activity enhancer
- the basic skeleton is a disaccharide repeat structure consisting of HexA residues and GlcN residues.
- the HexA residue is preferably a GlcA residue or an IdoA residue.
- the cells are preferably bone marrow-derived mesenchymal cells or osteoblasts.
- agents of the present invention are collectively referred to as "agents of the present invention”.
- the agent of the present invention is extremely useful because GAG, which is an active ingredient, exerts an excellent effect by directly acting on hard tissue formation, cell differentiation promotion or cell ALP activity enhancement. It is.
- Active ingredient of the agent of the present invention contains GAG or a salt thereof having a sulfate group and having the following properties (1) and (2) as an active ingredient.
- the basic skeleton is a disaccharide repeat structure consisting of HexA residues and GlcN residues.
- the HexA residue is not particularly limited, but is preferably a GlcA residue or an IdoA residue.
- the GlcA residue is preferably a D-GlcA residue.
- the IdoA residue is preferably a L IdoA residue.
- the GlcN residue is not particularly limited, but is preferably a D-GlcN residue.
- the amino group of the GlcN residue may be acetylated.
- the GlcN residue part is N-acetylylcosamine (GlcNAc).
- GAG which is an active ingredient of the agent of the present invention, has a basic skeleton of such a disaccharide repeating structure consisting of HexA residue and GlcN residue. That is, when the HexA residue is “a” and the GlcN residue is “b”, GAG, which is an active ingredient of the agent of the present invention, is “a-b-a-b” or “b-a-b”. “A” also means basic skeletal power. “-” Indicates a glycosidic bond. When a is a GlcA residue, the glycosidic bond (ab) between a and b is preferably a ⁇ 1 ⁇ 4 bond.
- the glycosidic bond (a ⁇ b) between a and b is preferably an ⁇ 1 ⁇ 4 bond.
- the glycosidic bond between b and a (b ⁇ a) is preferably an ⁇ 1 ⁇ 4 bond.
- Such GAG which is an active ingredient of the agent of the present invention, is characterized by retaining a sulfate group.
- the position at which the sulfate group can be retained in the basic skeleton is not particularly limited, but is selected from the hydroxyl group at the 2-position of the HexA residue, the hydroxyl group at the 6-position of the GlcN residue and the amino group at the 2-position of the GlcN residue. One or more positions are preferred. Examples of GAG in which a sulfate group is held at such a position include Hep and HS.
- the hydroxyl group at the 2-position of the HexA residue, the hydroxyl group at the 6-position of the GlcN residue, and the 2-position of the GlcN residue in the basic skeleton described above There must be no sulfate group at any one or less of the amino groups. It is important. Therefore, as an active ingredient of the agent of the present invention, for example, Hep (hydroxyl group at the 2-position of HexA residue, hydroxyl group at the 6-position of GlcN residue, and amino group at the 2-position of GlcN residue are sulfated. Groups are retained), 2DSH, 6DSH, NDSH, etc.
- GAG is a polysaccharide (polymer), and the size of all GAG molecules present in a fraction of a GAG, the composition and arrangement of constituent sugars, the position of sulfate groups, the number of sulfate groups, etc. are completely present. It goes without saying that it is technical common sense in this technical field that it is virtually impossible to be the same.
- the origin of GAG that is an active ingredient of the agent of the present invention is not particularly limited, and natural products that may be used as they are are processed using chemical methods, enzymatic methods, or other methods. Use a chemically synthesized material.
- Hep when Hep is used as an active ingredient of the agent of the present invention, Hep obtained from a natural product or the like by a known method may be used as it is. Hep already sold as a pharmaceutical or reagent may be used. It may be used. Alternatively, a method in which the degree of sulfuric acid in HS is increased by a known chemical method, enzymatic method, or other methods to the same extent as Hep may be used.
- the term “Hep” in the application documents of this application includes not only pure Hep but also what is recognized as substantially the same as Hep.
- the size (weight average molecular weight) of GAG that can be used as an active ingredient of the agent of the present invention is as long as it is recognized as "polysaccharide" by those skilled in the art. Although not particularly limited, for example, those having a weight average molecular weight of about 5000 to 20000 are preferred, and those having a weight average molecular weight of about 10,000 to 15000 are preferred.
- the weight average molecular weight can be measured according to the method described in WOOO / 06608.
- the purity of GAG as an active ingredient of the agent of the present invention is not particularly limited, and can be appropriately selected according to the scene and purpose of application of the agent of the present invention.
- the agent of the present invention when the agent of the present invention is applied to a subject requiring high sterility or cleanliness, such as a tissue in a living body or cells cultured under aseptic conditions, it is purified to a high purity, It is desirable to use a substance that does not substantially contain a substance that is not scientifically or culturically acceptable (for example, endotoxin or microorganism).
- a substance that is not scientifically or culturically acceptable for example, endotoxin or microorganism.
- the agent of the present invention when the agent of the present invention is applied to a subject whose sterility or cleanliness is not so much required, the degree of purification may be appropriately reduced according to the situation and purpose.
- the "GAG salt” is not particularly limited, and can be appropriately selected depending on the scene and purpose of application of the agent of the present invention.
- a medically acceptable salt may be selected.
- the salt of GAG include alkali metal salts (sodium salts, lithium salts, potassium salts, etc.), alkaline earth metal salts, salts with inorganic bases such as ammonium salts, diethanolamine salts, cyclohexylamine salts, amino acid salts, etc. And salts with organic bases. Of these, sodium salts are preferred.
- the agent of the present invention may contain only one type of the GAG or a salt thereof as an active ingredient, or may contain a plurality of types of the GAG or a salt thereof as an active ingredient.
- GAG or its salt By using such GAG or its salt, it can be set as the formation accelerator of this invention, this differentiation promoter, and this activity enhancer which have the outstanding effect.
- the dosage form of the agent of the present invention contains the aforementioned GAG or a salt thereof, and has an action of promoting hard tissue formation, an action of promoting differentiation of cells, and an action of enhancing ALP activity of cells by parentheses. It is not particularly limited as long as it is exhibited, and can be appropriately selected according to the scene and purpose of applying the agent of the present invention.
- the agent of the present invention may be a powder, granule or other solid agent as a solution-state agent containing the GAG or a salt thereof.
- a solution agent when it is provided as a solution agent, it may be provided in a frozen state or as a solution, or it may be provided as a dissolving agent at the time of use.
- the concentration of the GAG or a salt thereof in the agent of the present invention is not particularly limited, and can be appropriately selected according to the scene or purpose of application of the agent of the present invention.
- Formulation of the agent of the present invention can be performed by a known method.
- other drug-removing components such as an isotonic agent, a corrigent, a preservative, a pH adjuster, a soothing agent, a colorant, an excipient, a binder, a lubricant, and a disintegrant can be blended.
- the agent of the present invention can be a hard tissue formation promoter, a cell differentiation promoter, or a cell ALP activity enhancer.
- the “hard tissue” is preferably bone. “Promotion of hard tissue formation” includes the significance of promoting calcification.
- the “cell” may be a bone marrow-derived mesenchymal cell or an osteoblast. preferable.
- the agent of the present invention may be a single agent intended for one of these uses, or a single agent intended for a plurality of these uses.
- the agent of the present invention can be administered or administered as long as the GAG or its salt molecule, which is the active ingredient, is in contact with the target biological tissue or cell.
- the method of application is not limited.
- the agent of the present invention can be used, for example, as a drug or reagent for the purpose of promoting hard tissue formation, promoting cell differentiation, or enhancing cell ALP activity.
- the dosage is determined according to the purpose of administration (prevention, maintenance (prevention of deterioration), reduction (improvement of symptoms) or treatment), type of disease, patient symptoms, gender, year For age, weight, administration site, administration method, etc. Therefore, although it should be set individually and is not particularly limited, approximately 0.03 mg / dose site to 3 mg / dose site as GAG or a salt thereof can be generally administered per adult.
- the agent of the present invention is used as a medicine
- the animal to which it is administered is not particularly limited, but a human who prefers a mammal preferred by a vertebrate is particularly preferred. In this case, according to each use, it can administer to the following animals, for example.
- a cell differentiation promoting agent When used as a cell differentiation promoting agent, it can be administered to an animal in a state where cell sorting is desired. In particular, it is preferably administered to an animal in a state where it is desired to separate bone marrow-derived mesenchymal stem cells, osteoblasts, and the like. Examples of the state in which such cell differentiation is desired are the same as in the case of the hard tissue formation promoter.
- the agent of the present invention when used as a reagent, its application target is not particularly limited, and examples thereof include tissues and cultured cells obtained by collecting vital force.
- the GAG or a salt thereof which is an active ingredient of the agent of the present invention, is fixed on the surface of an appropriate insoluble carrier or mixed with an appropriate material to promote hard tissue formation. It may also be used as a material for the purpose of promoting cell differentiation or enhancing cell ALP activity.
- insoluble carriers materials
- Such insoluble carriers have various shapes including beads, films, plates, monofilaments, non-woven fabrics, sponges, woven fabrics, knitted fabrics, short fibers, tubes, hollow fibers, fibers, idroxypatites, etc.
- it can be used as a medical composite material for implants, bone cements, bone fillers, root canal fillers, fracture plates, artificial joints, and the like.
- the agent of the present invention is applied as a material in the field of regenerative medicine. Say it with a word.
- the ST2 cell line, a mouse bone marrow-derived mesenchymal cell line, and the MC3T3-E1 cell line (both obtained from RIKEN CELL BANK), a mouse osteoblast-like cell line with a higher degree of differentiation, were used.
- these cells were all cultured under conditions of 37 ° C and 5% CO. Also these
- Antibiotics (final concentration 100 U / ml penicillin G, final concentration 100 g / ml streptomycin and final concentration 250 g / ml gentamicin) coexisted in the medium used for cell culture.
- Hyaluronic acid with a weight average molecular weight of 60,000 (hyaluronic acid 60,000)
- Hyaluronic acid with a weight average molecular weight of 350,000 (hyaluronic acid 350,000)
- Hyaluronic acid with a weight average molecular weight of 1.5 million (hyaluronic acid 1.5 million)
- Hyaluronic acid with a weight average molecular weight of 2.5 million (hyaluronic acid 2.5 million)
- NSH the amino group at position 2 of the GlcN residue in Hep is sulfated, and the hydroxyl group at position 2 of the HexA residue and the hydroxyl group at position 6 of the GlcN residue do not retain a sulfate group
- 6SH 6- O-sulfated Hep; 6-0- sulfated heparin; 6-position hydroxyl of GlcN residue The group is sulfated, and the hydroxyl group at position 2 of the HexA residue and the amino group at position 2 of the GlcN residue do not retain a sulfate group.
- CDS skin sulfate
- Hep, 2DSH, 6DSH and NDSH used here were analyzed by the "enzymatic disaccharide analysis method combining digestion with GAG degrading enzyme and high performance liquid chromatography" described in WO00 / 06608 ( table 1).
- DiHS-OS is 2-acetamido-2-deoxy-4-0- (4-deoxy- ⁇ -L-threo-hex-4-enopyranosyluronic acid)-D-glucose
- ⁇ DiHS- NS is 2-deoxy-2-sulfamino-4-0- (4-deoxy-a-L-threo-hex-4-enobilanosyluronic acid)-D-glucose
- ⁇ DiHS-6S is 2-acetamido-2- Deoxy-4-0- (4-deoxy-a-L-threo-hex- 4-enopyranosyluronic acid) -6-0-sulfo-D-glucose
- ⁇ Di HS-US is 2-acetamido -2-deoxy-4-0- (4-deoxy-2-0-sulfo-a-L-threo-hex-4-enopyranosyluronic acid)-D-glucose with ⁇ DiHS-di (6 ,
- weight average molecular weights were measured by the method described in WO00 / 06608, they were Hep force S 14000, 2DSH force 12000, 6DSH force 12500, and NDSH force S 11000.
- the FBS manufactured by EQUITECH-BIO was used.
- Cellular ALP activity was measured as an indicator of cell sorting.
- Cells were seeded on a 24-well culture plate at a rate of 5 ⁇ 10 3 cells / well, and the test sample was added to ⁇ MEM containing 2% FBS at a final concentration of 50 g / ml and cultured.
- the ALP activity was also measured in the first week by the following enzymatic chemistry method (Bessey-Lowry method).
- the cultured cells are washed with PBS, then disrupted for 40 seconds in 10 mM Tris-HCl buffer (pH 7.4, 500 1) using an ultrasonic homogenizer (Handy Sonic model UR-20P; manufactured by Tommy Industries) and stirred. did. Next, add this sample solution 25 1 to 125 ⁇ 1 of ALP buffer (0.1 M carbonate buffer pH 9.8, 6.7 mM p-diphenyl phosphate, 2 mM MgCl 2) at 37 ° C for 30 minutes.
- ALP buffer 0.1 M carbonate buffer pH 9.8, 6.7 mM p-diphenyl phosphate, 2 mM MgCl 2
- the reaction was terminated by adding 125 ⁇ l of 0.2N NaOH, and the absorbance at 405 nm was measured using a microplate reader (Model 550, Bio-Rad Laboratories). Calculated ALP activity is in units per cell. It showed in. In addition, a test sample was used as a control without adding a test sample.
- Figure 1 shows the results of adding various test samples to the MC3T3-E1 cell line. Separately, test samples (Hep, 2DSH, 6DSH, or NDSH) were added to the MC3 T3-E1 cell line.
- Fig. 2 shows the results, and
- Fig. 3 shows the results of adding the test sample (Hep, 2DSH, 6DSH, or NDSH) to the ST2 cell line.
- Hep, 2DSH, 6DSH and NDSH all enhanced the ALP activity of the MC3T3-El cell line.
- Hep was shown to be particularly effective for enhancing ALP activity in the MC3T3-E1 cell line.
- FIG. 4 shows the results of adding the test sample (Hep, 2DSH, 6DSH, or NDSH) to the MC3T3-E1 cell line, and the test sample (Hep, 2DSH, 6DSH, or NDSH) to the ST2 cell line. The results are shown in Fig. 5. The calculated ALP activity was expressed as a value per cell DNA weight.
- Hep, 2DSH, 6DSH and NDSH all enhanced the ALP activity of the MC3T3-E1 cell line under serum-free conditions.
- Hep, 2DSH and NDSH were shown to be particularly effective for enhancing ALP activity in the MC3T3-E1 cell line.
- Hep, 2DSH, 6DSH and NDSH are all ST2 under serum-free conditions. ALP activity of the cell line was enhanced. In particular, 2DSH was shown to be particularly effective in enhancing ALP activity in the ST2 cell line.
- Hep, 2DSH, 6DSH and NDSH all enhance the ALP activity of bone marrow-derived mesenchymal cells and osteoblasts under serum-free conditions.
- Hep in which a sulfate group is held at two or more positions selected from the group consisting of 2-position, 6-position and N-position in Hep skeleton, has various factors contained in serum. This indicates that it acts on these cells directly without intervening to enhance ALP activity.
- Hep, 2DSH and NDSH particularly effectively enhanced ALP activity of osteoblasts, and 2DSH particularly effectively enhanced ALP activity of bone marrow-derived mesenchymal cells.
- alizarin staining (AL Z staining; stained red) in accordance with a conventional method one week after the cell reaches confluence Calcification was promoted so much).
- the cells were washed with PBS, fixed with 10 neutral buffered formalin, and stained with 0.01 Alizarin Red S (Wako Pure Chemical Industries).
- a hydroxyapatite block (size: length 3 mm x width 2 mm x height 2 mm; absorbs about 0.1 ml of solution) is immersed in a test sample with a concentration of 3 mg / ml for 30 minutes. Used for transplantation.
- test sample containing no test sample and using PBS was used as a control.
- 2DSH and NDSH are bone marrow-derived mesenchymal cells It has been shown to be particularly effective for increasing ALP activity in bone and osteoblasts and forming bone (hard tissue).
- Purified water was added to the above ingredients to make an lg cream.
- hydroxypropyl cellulose was granulated After a 30mg followed by kneading example mosquitoes ⁇ a solution in methanol (hydroxypropylcellulose 10 weight 0/0). This was extruded through a screen with a diameter of 0.8 mm to form granules, dried, and then added with 15 mg of magnesium stearate and tableted in 200 mg to obtain tablets.
- (A) and (B) were used as one set to produce an injection for dissolution at the time of use.
- (A) can be dissolved in (B) and used.
- the agent of the present invention can be used as a drug or reagent for the purpose of promoting hard tissue formation, promoting cell differentiation, enhancing cellular ALP activity, and the like.
- Fig. 1 is a graph showing enhancement of ALP activity in MC3T3-E1 cells.
- FIG. 2 is a graph showing enhancement of ALP activity in MC3T3-E1 cells.
- FIG. 3 is a graph showing enhancement of ALP activity in ST2 cells.
- FIG. 4 shows enhancement of ALP activity in MC3T3-E1 cells under serum-free conditions.
- FIG. 5 is a graph showing enhancement of ALP activity in ST2 cells under serum-free conditions.
- FIG. 6 is a diagram showing a ratio of an area of a portion where a bone (hard tissue) is formed.
- FIG. 7 is a diagram showing the ratio of the area of a portion where a bone (hard tissue) is formed.
- FIG. 8 is a graph showing the influence on the proliferation of MC3T3-E1 cells.
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Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
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JP2007508162A JP5153324B2 (ja) | 2005-03-14 | 2006-03-14 | 硬組織形成促進剤 |
US11/908,614 US20090012278A1 (en) | 2005-03-14 | 2006-03-14 | Promoter For Hard Tissue Formation |
EP06729086.6A EP1859803B1 (en) | 2005-03-14 | 2006-03-14 | Promoter for hard tissue formation |
US13/196,713 US8604003B2 (en) | 2005-03-14 | 2011-08-02 | Promoter for hard tissue formation |
Applications Claiming Priority (4)
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JP2005071023 | 2005-03-14 | ||
JP2005-071023 | 2005-03-14 | ||
JP2005-176311 | 2005-06-16 | ||
JP2005176311 | 2005-06-16 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
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US11/908,614 A-371-Of-International US20090012278A1 (en) | 2005-03-14 | 2006-03-14 | Promoter For Hard Tissue Formation |
US13/196,713 Division US8604003B2 (en) | 2005-03-14 | 2011-08-02 | Promoter for hard tissue formation |
Publications (1)
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WO2006098332A1 true WO2006098332A1 (ja) | 2006-09-21 |
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US (2) | US20090012278A1 (ja) |
EP (1) | EP1859803B1 (ja) |
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JP2016520697A (ja) * | 2013-05-16 | 2016-07-14 | エージェンシー フォー サイエンス,テクノロジー アンド リサーチ | ヘパラン硫酸 |
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ES2327480B1 (es) * | 2007-06-15 | 2010-08-10 | Bioiberica, S.A. | "disacaridos para el tratamiento de tendones, ligamentos y huesos". |
WO2017027474A1 (en) * | 2015-08-10 | 2017-02-16 | The Trustees Of Princetion University | Nano-encapsulation using gras materials and applications thereof |
TWI769176B (zh) * | 2016-09-07 | 2022-07-01 | 瑞瑟勒綜合技術協會 | 生合成肝素 |
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JP4035628B2 (ja) * | 2001-10-09 | 2008-01-23 | 生化学工業株式会社 | ギャップ機能亢進剤 |
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2006
- 2006-03-14 WO PCT/JP2006/305052 patent/WO2006098332A1/ja active Application Filing
- 2006-03-14 US US11/908,614 patent/US20090012278A1/en not_active Abandoned
- 2006-03-14 EP EP06729086.6A patent/EP1859803B1/en not_active Not-in-force
- 2006-03-14 JP JP2007508162A patent/JP5153324B2/ja not_active Expired - Fee Related
-
2011
- 2011-08-02 US US13/196,713 patent/US8604003B2/en not_active Expired - Fee Related
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Cited By (1)
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JP2016520697A (ja) * | 2013-05-16 | 2016-07-14 | エージェンシー フォー サイエンス,テクノロジー アンド リサーチ | ヘパラン硫酸 |
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US20090012278A1 (en) | 2009-01-08 |
EP1859803B1 (en) | 2016-10-19 |
JPWO2006098332A1 (ja) | 2008-08-21 |
JP5153324B2 (ja) | 2013-02-27 |
EP1859803A1 (en) | 2007-11-28 |
US8604003B2 (en) | 2013-12-10 |
EP1859803A4 (en) | 2012-02-08 |
US20110288048A1 (en) | 2011-11-24 |
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