WO2003101471A1 - Calcium-containing composition from bone tissue - Google Patents

Calcium-containing composition from bone tissue Download PDF

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Publication number
WO2003101471A1
WO2003101471A1 PCT/NL2003/000400 NL0300400W WO03101471A1 WO 2003101471 A1 WO2003101471 A1 WO 2003101471A1 NL 0300400 W NL0300400 W NL 0300400W WO 03101471 A1 WO03101471 A1 WO 03101471A1
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WO
WIPO (PCT)
Prior art keywords
calcium
bone tissue
containing composition
bones
process according
Prior art date
Application number
PCT/NL2003/000400
Other languages
French (fr)
Inventor
Dick Teune
Johannes Adrianus Hage
Original Assignee
Gelatine Smits Beheer B.V.
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 Gelatine Smits Beheer B.V. filed Critical Gelatine Smits Beheer B.V.
Priority to EP03741622A priority Critical patent/EP1509238A1/en
Priority to AU2003273569A priority patent/AU2003273569A1/en
Publication of WO2003101471A1 publication Critical patent/WO2003101471A1/en

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Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/32Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00Drugs for skeletal disorders
    • A61P19/08Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis

Definitions

  • the present invention relates to a calcium-containing composition obtained from animal bone tissue for treating disorders connected with the growth or healing of bones or bone tissue-
  • a composition is known from EP 0 255 565.
  • the known composition has a total protein content of at least 30.6 wt. %, the collagen content being at least 23.4 wt. %.
  • the known composition has a calcium content of maximally 23.4 wt. %, and the particle size of the composition can be 300 ⁇ m.
  • the calcium-containing compositions known from WO 94/26283 are based on organic or inorganic calcium salts, for example in the form of aragonite or calcite, possibly obtained from external skeletons of corals and crustaceans.
  • disorders connected with the growth or healing of bones or bone tissue is understood to mean not only disorders relating to an insufficient mineralisation or a decreasing mineralisation (decalcification), such as a dysplasia, an osteodystrophy, a paradontal disease or osteoporosis, but also the local healing of bone fractures.
  • a calcium-containing composition having a matrix structure of natural bone tissue and a very high biological availability for a better and more efficient treatment of such disorders.
  • Known calcium-containing composition based on calcium salts such as calcium chloride and calcium carbonate lack the phosphate ion that is essential for the building up of bone tissue.
  • calcium-containing compositions obtained from the exoskeleton of lower animals does not have the suitable matrix structure and the required high biological availability. According to the present invention, this need is met by a calcium-containing composition, which is characterised in that the composition is a calcium phosphate complex having a protein content of less than 25 wt. % and that the composition has a matrix of natural bone tissue.
  • composition according to the present invention is the fact that there is no risk of contaminations with, for example, BSE causing agents, in particular because of the low protein content (obtained by using the extraction method to be described hereinafter).
  • the protein content amounts to less than 10 wt. %.
  • the calcium content amounts to more than 30%.
  • the bone tissue may be obtained from various animal species, for example cattle, poultry or pigs.
  • the calcium phosphate complex is obtained from natural porcine bone tissue. Not only is the matrix structure of porcine bone tissue more similar to the matrix structure of human bone tissue, but in addition there is no danger of contamination with BSE causing agents when porcine bone tissue is used. Research has shown that the biological availability is higher if the calcium phosphate complex has been sufficiently reduced in size, and preferably the particle size of the obtained calcium complex is maximally 75 ⁇ m.
  • the present invention also relates to a process for preparing a calcium-containing composition starting from natural bone tissue, comprising the defatting and grinding of bones. Such a method is generally known.
  • the process according to the invention is characterized in that the bones, which may be crushed to form small pieces, are defatted at a temperature of less than 90 "C, after which proteins are extracted by first treating the bones with a liquid having an acidic pH value and subsequently treating the bones with steam, after which the bones are grinded.
  • the bones are dried with heated air following the defatting step and prior to the extracting step.
  • a very good extraction is obtained if the liquid used in the extracting step has a pH value of 2-5.
  • a process step which leads to good extraction results and which is nevertheless sufficiently mild to maintain a natural matrix structure of bone tissue comprises treating with steam under pressure of at least 3 bar for a period of at least 20 minutes.
  • the bones are washed with water having a temperature of maximally 80 °C following the treatment with steam and before the grinding step.
  • the present invention also relates to a pharmaceutical preparation on the basis of a calcium-containing composition for treating disorders connected with the growth or healing of bones or bone tissue, which preparation comprises at least one active ingredient and one auxiliary substance, wherein the preparation contains a calcium- containing composition according to the present invention as the active ingredient.
  • the present invention relates to a pharmaceutical preparation on the basi's of a calcium-containing composition for treating osteoporosis, which comprises at least one active ingredient and one auxiliary substance, wherein the preparation contains a calcium- containing composition according to the present invention as the active ingredient.
  • An auxiliary substance may be added for the purpose of improving the running properties; in this connection, good results have been obtained with carboxylmethyl cellulose as the auxiliary substance.
  • the calcium-containing composition incorporated in a pharmaceutical preparation can be administered, for example orally, to a human or animal patient, in amounts and at a frequency to be determined by the physician in attendance, inter alia depending on the gravity of the disorder in question or the extent of the required healing, the age and the body weight of the patient.
  • a soft capsule containing an amount of a calcium-containing composition corresponding to 100 mg of calcium ions may be orally administered to the patient, four times a day.
  • Alternative dosage forms are hard capsules, each containing 125 mg of calcium ions or granulates containing 30 wt.% of calcium ions.
  • Administration in liquid form for example in the form of a suspension in a buffered citrate solution, is also suitable.
  • porcine bones An amount of 100 g of porcine bones was crushed into pieces of maximally 2 cm.
  • the porcine bones were defatted with water at 80 ° C, followed by the removal of the soft material and the finest bone particles. Finally, the defatted bone material was dried with heated air.
  • the defatted and dried bone material was introduced into an autoclave for carrying out the extraction process and was treated with a solution of 0.1 M sulphuric acid, for a period of 1 hour. The thus obained maceration liquid was removed. Subsequently, the bone material was saturated with steam, after which the pressure was increased to 3 bar, which pressure was maintained for 20 minutes. After expansion, the treated bone material was washed with water having a temperature of 60 "C. This extraction procedure was repeated seven times.
  • the defatted and extracted bone material was dried with heated air (60 °C) and finally ground in a ball mill to a particle size of maximally 75 ⁇ m.
  • the fat content was 1%.
  • the calcium content was 32%.
  • the calcium composition that was eventually obtained was subjected to a test regarding the biological availability thereof.
  • the calcium composition was stirred in a slightly acidic cyclic acid solution at 65 ° C for one hour, and was subsequently subjected to filtration.
  • the calcium content of the filtrate was determined.
  • the determination of the biological availability or the capacity of biological incorporation was carried out substantially as described in Dellaert B.N. et al , Netherlands Journal of Agricultural Science, 38, pp. 555-566 (1990). The biological availability was shown out to be 97.2%.
  • Example 2 The determination of the biological availability or the capacity of biological incorporation was carried out substantially as described in Dellaert B.N. et al , Netherlands Journal of Agricultural Science, 38, pp. 555-566
  • Example 2 Using the calcium-containing composition based on a calcium phosphate complex obtained from porcine bone tissue, obtained as described in Example 1, a pharmaceutical preparation suitable for treating disorders connected with the growth or healing of bones or bone tissue was prepared. An amount of 99 g of the calcium-containing composition obtained by using the method described in Example 1 was mixed with an amount of 1 gram of carboxylmethylcellulose.
  • the pharmaceutical preparation was administered orally in the form of hard capsules (each containing an amount of calcium- containing composition corresponding to 125 mg of calcium ions) to 50 adult female volunteers, using a dosing scheme of four times one capsule, distributed over the day. Two other groups of 50 volunteers each were given a known calcium composition and a placebo, respectively. The test was carried out as a double-blind test. The test results showed an increased biological availability and an excellent incorporation of calcium obtained by using the calcium composition according to the invention.
  • the pharmaceutical preparation as prepared was shown to be suitable for helping to treat disorders connected with the growth of bones or bone tissue, such as, by way of example, osteoporosis.

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  • Health & Medical Sciences (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Rheumatology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical & Material Sciences (AREA)
  • Cell Biology (AREA)
  • Biomedical Technology (AREA)
  • Epidemiology (AREA)
  • Virology (AREA)
  • Immunology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The present invention relates to a calcium-containing composition obtained from animal bone tissue for treating disorders connected with the growth or healing of bones or bone tissue, wherein the composition is a calcium phosphate complex having a protein content of less than 25 wt. % and wherein the composition has a matrix of natural bone tissue. The present invention furthermore relates to a process for preparing a calcium-containing composition of natural bone tissue and a pharmaceutical preparation on the basis of said composition.

Description

CALCIUM-CONTAINING COMPOSITION FROM BONE TISSUE
DESCRIPTION The present invention relates to a calcium-containing composition obtained from animal bone tissue for treating disorders connected with the growth or healing of bones or bone tissue- Such a composition is known from EP 0 255 565. The known composition has a total protein content of at least 30.6 wt. %, the collagen content being at least 23.4 wt. %. Moreover, the known composition has a calcium content of maximally 23.4 wt. %, and the particle size of the composition can be 300 μm.
The calcium-containing compositions known from WO 94/26283 are based on organic or inorganic calcium salts, for example in the form of aragonite or calcite, possibly obtained from external skeletons of corals and crustaceans.
In the context of the present invention, the phrase "disorders connected with the growth or healing of bones or bone tissue" is understood to mean not only disorders relating to an insufficient mineralisation or a decreasing mineralisation (decalcification), such as a dysplasia, an osteodystrophy, a paradontal disease or osteoporosis, but also the local healing of bone fractures.
In the state of the art, there is a need for a calcium- containing composition having a matrix structure of natural bone tissue and a very high biological availability for a better and more efficient treatment of such disorders. Known calcium-containing composition based on calcium salts such as calcium chloride and calcium carbonate lack the phosphate ion that is essential for the building up of bone tissue. Furthermore, it has become apparent that calcium-containing compositions obtained from the exoskeleton of lower animals does not have the suitable matrix structure and the required high biological availability. According to the present invention, this need is met by a calcium-containing composition, which is characterised in that the composition is a calcium phosphate complex having a protein content of less than 25 wt. % and that the composition has a matrix of natural bone tissue.
An additional advantage of the composition according to the present invention is the fact that there is no risk of contaminations with, for example, BSE causing agents, in particular because of the low protein content (obtained by using the extraction method to be described hereinafter). In this connection it is preferable that the protein content amounts to less than 10 wt. %.
Preferably, the calcium content amounts to more than 30%. In principle, the bone tissue may be obtained from various animal species, for example cattle, poultry or pigs. Preferably, however, the calcium phosphate complex is obtained from natural porcine bone tissue. Not only is the matrix structure of porcine bone tissue more similar to the matrix structure of human bone tissue, but in addition there is no danger of contamination with BSE causing agents when porcine bone tissue is used. Research has shown that the biological availability is higher if the calcium phosphate complex has been sufficiently reduced in size, and preferably the particle size of the obtained calcium complex is maximally 75 μm.
The present invention also relates to a process for preparing a calcium-containing composition starting from natural bone tissue, comprising the defatting and grinding of bones. Such a method is generally known.
In order to prepare calcium-containing composition that has a matrix structure of natural bone tissue and a high biological availability and, in addition, and moreover has a low protein content, special process steps are required, during which mild conditions have to be used in order to maintain the natural matrix structure as much as possible.
The process according to the invention is characterized in that the bones, which may be crushed to form small pieces, are defatted at a temperature of less than 90 "C, after which proteins are extracted by first treating the bones with a liquid having an acidic pH value and subsequently treating the bones with steam, after which the bones are grinded.
Preferably, the bones are dried with heated air following the defatting step and prior to the extracting step. A very good extraction is obtained if the liquid used in the extracting step has a pH value of 2-5. A process step which leads to good extraction results and which is nevertheless sufficiently mild to maintain a natural matrix structure of bone tissue comprises treating with steam under pressure of at least 3 bar for a period of at least 20 minutes. Preferably, the bones are washed with water having a temperature of maximally 80 °C following the treatment with steam and before the grinding step.
In order to ensure a satisfactory extraction when using the present process conditions, it is advisable to carry out the extraction procedure a few times, preferably at least three times, and very good results are obtained if the procedure is repeated at least seven times.
The present invention also relates to a pharmaceutical preparation on the basis of a calcium-containing composition for treating disorders connected with the growth or healing of bones or bone tissue, which preparation comprises at least one active ingredient and one auxiliary substance, wherein the preparation contains a calcium- containing composition according to the present invention as the active ingredient. Finally, the present invention relates to a pharmaceutical preparation on the basi's of a calcium-containing composition for treating osteoporosis, which comprises at least one active ingredient and one auxiliary substance, wherein the preparation contains a calcium- containing composition according to the present invention as the active ingredient. An auxiliary substance may be added for the purpose of improving the running properties; in this connection, good results have been obtained with carboxylmethyl cellulose as the auxiliary substance. The calcium-containing composition incorporated in a pharmaceutical preparation can be administered, for example orally, to a human or animal patient, in amounts and at a frequency to be determined by the physician in attendance, inter alia depending on the gravity of the disorder in question or the extent of the required healing, the age and the body weight of the patient. For example, in the case of a human patient, in particular a female patient, a soft capsule containing an amount of a calcium-containing composition corresponding to 100 mg of calcium ions may be orally administered to the patient, four times a day. Alternative dosage forms are hard capsules, each containing 125 mg of calcium ions or granulates containing 30 wt.% of calcium ions. Administration in liquid form, for example in the form of a suspension in a buffered citrate solution, is also suitable.
The invention will now be illustrated in more detail by means of the following examples. Example 1.
An amount of 100 g of porcine bones was crushed into pieces of maximally 2 cm. The porcine bones were defatted with water at 80 °C, followed by the removal of the soft material and the finest bone particles. Finally, the defatted bone material was dried with heated air. The defatted and dried bone material was introduced into an autoclave for carrying out the extraction process and was treated with a solution of 0.1 M sulphuric acid, for a period of 1 hour. The thus obained maceration liquid was removed. Subsequently, the bone material was saturated with steam, after which the pressure was increased to 3 bar, which pressure was maintained for 20 minutes. After expansion, the treated bone material was washed with water having a temperature of 60 "C. This extraction procedure was repeated seven times.
The defatted and extracted bone material was dried with heated air (60 °C) and finally ground in a ball mill to a particle size of maximally 75 μm. The fat content was 1%. The calcium content was 32%. The calcium composition that was eventually obtained was subjected to a test regarding the biological availability thereof. The calcium composition was stirred in a slightly acidic cyclic acid solution at 65 °C for one hour, and was subsequently subjected to filtration. The calcium content of the filtrate was determined. The determination of the biological availability or the capacity of biological incorporation was carried out substantially as described in Dellaert B.N. et al , Netherlands Journal of Agricultural Science, 38, pp. 555-566 (1990). The biological availability was shown out to be 97.2%. Example 2. Using the calcium-containing composition based on a calcium phosphate complex obtained from porcine bone tissue, obtained as described in Example 1, a pharmaceutical preparation suitable for treating disorders connected with the growth or healing of bones or bone tissue was prepared. An amount of 99 g of the calcium-containing composition obtained by using the method described in Example 1 was mixed with an amount of 1 gram of carboxylmethylcellulose.
The pharmaceutical preparation was administered orally in the form of hard capsules (each containing an amount of calcium- containing composition corresponding to 125 mg of calcium ions) to 50 adult female volunteers, using a dosing scheme of four times one capsule, distributed over the day. Two other groups of 50 volunteers each were given a known calcium composition and a placebo, respectively. The test was carried out as a double-blind test. The test results showed an increased biological availability and an excellent incorporation of calcium obtained by using the calcium composition according to the invention.
The pharmaceutical preparation as prepared was shown to be suitable for helping to treat disorders connected with the growth of bones or bone tissue, such as, by way of example, osteoporosis.

Claims

1. Calcium-containing composition obtained from animal bone tissue for treating disorders connected with the growth or healing of bones or bone tissue, characterised in that the composition is a calcium phosphate complex having a protein content of less than 25 wt. %, and that the composition has a matrix of natural bone tissue.
2. Calcium-containing composition according to claim 1, characterised in that the protein content is less than 10%.
3. Calcium-containing composition according to claim 1 or 2, characterised in that the calcium content is more than 30%.
4. Calcium-containing composition according to any one of the claims 1 - 3, characterised in that the calcium phosphate complex is obtained from natural porcine bone tissue.
5. Calcium-containing composition according to any one of the claims 1 - 4, characterised in that the particle size of the obtained calcium phosphate complex is maximally 75 μm.
6. Process for preparing a calcium-containing composition starting from natural bone tissue, comprising the defatting and grinding of bones, characterised in that the bones, which may be crushed to form small pieces, are defatted at a temperature of less than 90 °C, after which proteins are extracted by first treating the bones with a liquid having an acidic pH value and subsequently treating the bones with steam, after which the bones are grinded.
7. Process according to claim 6, characterised in that, following the defatting step and prior to the extracting step, the bones are dried with heated air.
8. Process according to any one of the claims 6 or 7, characterised in that the liquid used for the extraction has a pH value of 2-5.
9. Process according to any one of the claims 6 - 8, characterised in that the treatment with steam is carried out for a period of at least 20 minutes and under pressure of at least 3 bar.
10. Process according to any one of the claims 6 - 9, characterised in that, after the treatment with steam and before the grinding step, the bones are washed with water having a temperature of maximally 80 °C.
11. Process according to any one of the claims 6 - 10, wherein the extraction procedure is carried out at least three times.
12. Process according to claim 11, wherein the extraction procedure is carried out at least seven times.
13. Pharmaceutical preparation on the basis of a calcium- containing composition obtained from animal bone tissue for treating disorders connected with the growth or healing of bones or bone tissue, which comprises at least one active ingredient and one auxiliary substance, characterised in that the preparation contains a calcium- containing composition according to any one of the claims 1 - 5 as the active ingredient.
14. Pharmaceutical preparation on the basis of a calcium- containing composition obtained from animal bone tissue for treating osteoporosis, which preparation comprises at least one active ingredient and one auxiliary substance, characterised in that the preparation contains a calcium-containing composition according to any one of the claims 1 - 5 as the active ingredient.
15. Pharmaceutical preparation according to claim 13 or claim 14, characterised in that the auxiliary substance is carboxylmethyl - cellulose.
PCT/NL2003/000400 2002-05-31 2003-05-28 Calcium-containing composition from bone tissue WO2003101471A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP03741622A EP1509238A1 (en) 2002-05-31 2003-05-28 Calcium-containing composition from bone tissue
AU2003273569A AU2003273569A1 (en) 2002-05-31 2003-05-28 Calcium-containing composition from bone tissue

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
NL1020729A NL1020729C2 (en) 2002-05-31 2002-05-31 Calcium-containing composition, method for preparing thereof, and pharmaceutical composition based on the composition.
NL1020729 2002-05-31

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WO2003101471A1 true WO2003101471A1 (en) 2003-12-11

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AU (1) AU2003273569A1 (en)
NL (1) NL1020729C2 (en)
WO (1) WO2003101471A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018220141A1 (en) 2017-06-01 2018-12-06 Fundació Institut De Bioenginyeria De Catalunya (Ibec) Controllable ion release calcium particles, method for producing same and use thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0255565A2 (en) * 1986-08-05 1988-02-10 Robapharm AG Composition for stimulating chondrocytes and osteoblasts (ossein hydroxyapatite compound), method for its production and pharmaceutical products containing said composition)

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0255565A2 (en) * 1986-08-05 1988-02-10 Robapharm AG Composition for stimulating chondrocytes and osteoblasts (ossein hydroxyapatite compound), method for its production and pharmaceutical products containing said composition)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018220141A1 (en) 2017-06-01 2018-12-06 Fundació Institut De Bioenginyeria De Catalunya (Ibec) Controllable ion release calcium particles, method for producing same and use thereof
US11491263B2 (en) 2017-06-01 2022-11-08 Fundacio Institut De Bioenginyeria De Catalunya (Ibec) Controllable ion release calcium particles, method for producing same and use thereof

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NL1020729C2 (en) 2003-12-02
AU2003273569A1 (en) 2003-12-19
EP1509238A1 (en) 2005-03-02

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