JP2005281186A - Wound healing agent, agent for amelioration, promotion of healing or prevention of injure or the like of tendon or ligament, functional food and medicine - Google Patents
Wound healing agent, agent for amelioration, promotion of healing or prevention of injure or the like of tendon or ligament, functional food and medicine Download PDFInfo
- Publication number
- JP2005281186A JP2005281186A JP2004097205A JP2004097205A JP2005281186A JP 2005281186 A JP2005281186 A JP 2005281186A JP 2004097205 A JP2004097205 A JP 2004097205A JP 2004097205 A JP2004097205 A JP 2004097205A JP 2005281186 A JP2005281186 A JP 2005281186A
- Authority
- JP
- Japan
- Prior art keywords
- agent
- wound healing
- tendon
- ligament
- gly
- Prior art date
- Legal status (The legal status 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 status listed.)
- Pending
Links
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
本発明は、創傷治癒剤、腱の損傷または断裂に対する改善、治癒促進または予防剤、腱の改善または強化剤、靱帯の損傷または断裂に対する改善、治癒促進または予防剤、靱帯の改善または強化剤、競走馬の屈腱の炎症、損傷または断裂に対する改善、治癒促進または予防剤、競走馬の屈腱の改善または強化剤、競走馬の靱帯の損傷または断裂に対する改善、治癒促進または予防剤、競走馬の靱帯の改善または強化剤、機能性食品ならびに医薬品に関するものである。 The present invention relates to a wound healing agent, improvement against tendon damage or tear, healing promotion or prevention agent, tendon improvement or strengthening agent, improvement against ligament damage or tear, healing promotion or prevention agent, ligament improvement or strengthening agent, Racehorse flexor tendon inflammation, damage or rupture improvement, healing promotion or prevention agent, racehorse flexion tendon improvement or strengthening agent, racehorse ligament damage or rupture improvement, healing promotion or prevention agent, racehorse ligament It relates to improving or enhancing agents, functional foods and pharmaceuticals.
コラーゲンは体を構成するタンパク質の約３０％を占めており、皮膚、血管、内臓および骨組織などいたるところに存在している。皮膚では真皮の７０％がコラーゲンからできており、個々の筋肉を包む筋膜もコラーゲンで構成されている。さらに細胞本体の外側を取り囲み、細胞集団を支える足場となっている細胞マトリックスの主要成分がやはりコラーゲンであることが知られており、旧来から、コラーゲンやコラーゲンを加熱変性させたゼラチン、およびそれらの加水分解物は化粧品及び食品向けの原材料、更に医薬品向けの生体機能性材料として幅広く利用されてきた。 Collagen accounts for about 30% of the protein that makes up the body, and is present throughout the skin, blood vessels, viscera and bone tissue. In the skin, 70% of the dermis is made of collagen, and the fascia enclosing individual muscles is also made of collagen. Furthermore, it is known that the main component of the cell matrix that surrounds the outside of the cell body and serves as a scaffold for supporting the cell population is collagen, and from the past, collagen and collagen obtained by heat-denaturing collagen, and their Hydrolysates have been widely used as raw materials for cosmetics and foods, and further as biofunctional materials for pharmaceuticals.
そして、近年、コラーゲンタンパクおよびその分解物の経口摂取の薬理効果について多くの研究がなされるようになった。これまでに、主な効果として、骨粗鬆症の予防・改善に結びつく骨芽細胞増殖促進作用、骨強化作用（例えば、特許文献１，２参照。）、加齢に伴う生体組織の機能の低下を改善させる、生体組織の新陳代謝促進作用（例えば、特許文献３，４参照。）、皮膚代謝促進作用、皮膚賦活作用（例えば、特許文献４，５参照。）など、数多くの効果が明らかとなっている。 In recent years, many studies have been conducted on the pharmacological effects of oral intake of collagen proteins and their degradation products. So far, the main effects include osteoblast proliferation promotion action, bone strengthening action (for example, see Patent Documents 1 and 2), which leads to prevention / improvement of osteoporosis, and improvement of the deterioration of biological tissue function with aging. Numerous effects have been clarified, such as the metabolism promoting action of living tissues (see, for example, Patent Documents 3 and 4), the skin metabolism promoting action, and the skin activation action (see, for example, Patent Documents 4 and 5). .
さらに、本発明者らの研究により、コラーゲン成分あるいはゼラチン成分をコラゲナーゼ酵素を用いて特異的に分解して得られる、アミノ酸配列が Ｇｌｙ−Ｘ−Ｙのコラーゲントリペプチドが、従来のコラーゲン、ゼラチン、あるいはその加水分解物よりも速やかに効率よく消化吸収され、前記の機能をはじめとした種々の機能を発揮することが明らかとなり（特許文献６参照。）、少量の摂取でも効率よく速やかに消化吸収される、コラーゲン産生促進剤、機能性食品、医薬品、骨芽細胞または線維芽細胞の増殖促進剤、骨の強化または密度向上剤、生体組織の新陳代謝促進剤、皮膚賦活剤、皮膚または骨の加齢による機能低下の予防または改善剤、慢性リウマチ、変形関節症の改善剤および骨折治癒などの創傷治癒促進剤の提供が可能となった。 Furthermore, according to the study by the present inventors, a collagen tripeptide having an amino acid sequence of Gly-XY, which is obtained by specifically degrading a collagen component or a gelatin component using a collagenase enzyme, is obtained by using conventional collagen, gelatin, Or it becomes clear that it is digested and absorbed more quickly and efficiently than the hydrolyzate, and exhibits various functions including the above-mentioned functions (see Patent Document 6). Collagen production promoters, functional foods, pharmaceuticals, osteoblast or fibroblast growth promoters, bone strengthening or density improvers, biological tissue metabolism promoters, skin activators, skin or bone additives It is possible to provide agents for preventing or improving functional deterioration due to age, chronic rheumatism, osteoarthritis improving agents, and wound healing promoters such as fracture healing became.
主にスポーツ時に発症するアキレス腱断裂や靭帯損傷などの治癒には長期間要することが多く、復帰の遅れにより結果的に選手生命を絶たれてしまうアスリートも少なくない。また、人間だけでなく競走馬においてもヒトのアキレス腱に相当する屈腱部で発症する屈腱炎の問題が深刻化している。年間1,200頭の競走馬が屈腱炎を発症し、症状の重い馬は引退を余儀なくされ、また、回復の見込みがあっても競走復帰が大幅に遅れる一方、再発率も高く、競走馬にとって屈腱炎は不治の病と考えられている。このような背景から、腱断裂、靭帯損傷、屈腱炎などの治癒を促進する効果を有する創傷治癒剤、機能性食品、医薬品が切望されている。 Healing of Achilles tendon rupture and ligament injury that occurs mainly during sports often takes a long time, and many athletes lose their lives as a result of a delay in return. Moreover, the problem of flexor tendinitis that develops at the flexor tendon corresponding to the human Achilles tendon is becoming serious not only in humans but also in racehorses. 1,200 racehorses develop flexor tendinitis annually, severely ill horses are forced to retire, and even if they are expected to recover, the return to race is significantly delayed, but the recurrence rate is high, and tendonitis for racehorses Is considered an incurable disease. Under such circumstances, wound healing agents, functional foods, and pharmaceuticals having an effect of promoting healing of tendon rupture, ligament injury, flexor tendonitis and the like are eagerly desired.
本発明は、このような課題を解決するためになされたもので、新規で有用な創傷治癒剤、腱の損傷または断裂に対する改善、治癒促進または予防剤、腱の改善または強化剤、靱帯の損傷または断裂に対する改善、治癒促進または予防剤、靱帯の改善または強化剤、競走馬の屈腱の炎症、損傷または断裂に対する改善、治癒促進または予防剤、競走馬の屈腱の改善または強化剤、競走馬の靱帯の損傷または断裂に対する改善、治癒促進または予防剤、競走馬の靱帯の改善または強化剤、その創傷治癒剤を含む機能性食品ならびに医薬品を提供することを目的としている。 The present invention has been made to solve such problems, and is a novel and useful wound healing agent, improvement against tendon damage or rupture, healing promotion or prevention agent, tendon improvement or strengthening agent, ligament damage. Or improvement for rupture, healing or prevention agent, ligament improvement or strengthening agent, inflammation of racehorse flexor tendon, improvement for injury or tear, healing promotion or prevention agent, racehorse flexor tendon improvement or strengthening agent, racehorse An object of the present invention is to provide an improvement agent for ligament damage or rupture, an agent for promoting or preventing healing, an agent for improving or reinforcing a ligament of a racehorse, a functional food containing the wound healing agent, and a medicine.
本発明者らは、コラーゲン成分あるいはゼラチン成分をコラゲナーゼ酵素を用いて特異的に分解して得られる、アミノ酸配列がＧｌｙ−Ｘ−Ｙのコラーゲントリペプチドが、少量の摂取でも効率よく速やかに消化吸収され、コラーゲン産生促進、骨芽細胞または線維芽細胞の増殖促進、骨の強化または密度向上、生体組織の新陳代謝促進、皮膚賦活、皮膚または骨の加齢による機能低下の予防または改善、慢性リウマチ、変形関節症の改善および骨折治癒促進などの機能を有することを明らかにした（特許文献６参照。）。さらに、本発明者らは、そのコラーゲントリペプチドが腱断裂、靭帯損傷、屈腱炎などの治癒促進効果も有することを見出した。 The present inventors have been able to efficiently digest and absorb a collagen tripeptide having an amino acid sequence of Gly-XY, which is obtained by specifically degrading a collagen component or a gelatin component using a collagenase enzyme, even in a small amount. Promotes collagen production, promotes osteoblast or fibroblast proliferation, strengthens or increases bone density, promotes metabolism of living tissue, activates skin, prevents or improves functional deterioration due to aging of skin or bone, chronic rheumatism, It has been clarified that it has functions such as improving osteoarthritis and promoting healing of fractures (see Patent Document 6). Furthermore, the present inventors have found that the collagen tripeptide also has healing promoting effects such as tendon rupture, ligament damage, flexor tendinitis and the like.
即ち、本発明に係る創傷治癒剤は、コラーゲンまたはゼラチンをコラゲナーゼにより分解して得られる分解物であって、アミノ酸配列が（Ｇｌｙ−Ｘ−Ｙ）n（式中、Ｇｌｙはグリシン残基を表し、ＸおよびＹはグリシン以外の任意のアミノ酸残基を表し、ｎは正の整数を表す）で表されるペプチドを含むものを有効成分とすることを特徴とする。 That is, the wound healing agent according to the present invention is a degradation product obtained by degrading collagen or gelatin with collagenase, and the amino acid sequence is (Gly-XY) n (wherein Gly represents a glycine residue). , X and Y represent any amino acid residue other than glycine, and n represents a positive integer).
本発明に係る創傷治癒剤は、コラーゲンまたはゼラチンをコラゲナーゼにより分解して得られる分解物であって、アミノ酸配列が（Ｇｌｙ−Ｘ−Ｙ）n（式中、Ｇｌｙはグリシン残基を表し、ＸおよびＹはグリシン以外の任意のアミノ酸残基を表し、ｎは正の整数を表す）で表されるペプチドを有効成分とすることが好ましい。
本発明に係る創傷治癒剤で、前記分解物はアミノ酸配列がＧｌｙ−Ｘ−Ｙで表されるトリペプチドを５質量％以上含むことが好ましい。
The wound healing agent according to the present invention is a degradation product obtained by degrading collagen or gelatin with collagenase, and the amino acid sequence is (Gly-XY) n (wherein Gly represents a glycine residue, X And Y represents any amino acid residue other than glycine, and n represents a positive integer).
In the wound healing agent according to the present invention, the degradation product preferably contains 5% by mass or more of a tripeptide whose amino acid sequence is represented by Gly-XY.
さらに、本発明に係る創傷治癒剤は、コラーゲンまたはゼラチンをコラゲナーゼにより分解して得られる分解物であって、アミノ酸配列がＧｌｙ−Ｘ−Ｙ（式中、Ｇｌｙはグリシン残基を表し、ＸおよびＹはグリシン以外の任意のアミノ酸残基を表す）で表されるトリペプチドを有効成分とすることが好ましい。
本発明に係る創傷治癒剤は、創傷の治癒を促進し、早期回復に寄与する。
Further, the wound healing agent according to the present invention is a degradation product obtained by degrading collagen or gelatin with collagenase, and has an amino acid sequence of Gly-XY (where Gly represents a glycine residue, X and Y represents an arbitrary amino acid residue other than glycine).
The wound healing agent according to the present invention promotes wound healing and contributes to early recovery.
本発明に係る創傷治癒剤は、アスコルビン酸またはその誘導体が配合されていることが好ましい。この場合、創傷の治癒を促進する効果を高めることができる。 The wound healing agent according to the present invention preferably contains ascorbic acid or a derivative thereof. In this case, the effect of promoting wound healing can be enhanced.
本発明に係る腱の損傷または断裂に対する改善、治癒促進または予防剤は、前述の創傷治癒剤から成ることを特徴とする。本発明に係る腱の損傷または断裂に対する改善、治癒促進または予防剤は、腱の損傷または断裂を改善、治癒促進または予防する効果を有する。
本発明に係る腱の改善または強化剤は、前述の創傷治癒剤から成ることを特徴とする。本発明に係る腱の改善または強化剤は、腱を改善または強化する効果を有する。
The improvement, healing promotion or preventive agent for tendon damage or tear according to the present invention is characterized by comprising the aforementioned wound healing agent. The agent for improving, healing promoting or preventing tendon damage or tear according to the present invention has the effect of improving, promoting or preventing healing of tendon damage or tear.
The tendon improving or reinforcing agent according to the present invention is characterized by comprising the aforementioned wound healing agent. The tendon improving or reinforcing agent according to the present invention has an effect of improving or strengthening a tendon.
本発明に係る靱帯の損傷または断裂に対する改善、治癒促進または予防剤は、前述の創傷治癒剤から成ることを特徴とする。本発明に係る靱帯の損傷または断裂に対する改善、治癒促進または予防剤は、靱帯の損傷または断裂を改善、治癒促進または予防する効果を有する。
本発明に係る靱帯の改善または強化剤は、前述の創傷治癒剤から成ることを特徴とする。本発明に係る靱帯の改善または強化剤は、靱帯を改善または強化する効果を有する。
The agent for improving, healing or preventing ligament damage or rupture according to the present invention is characterized by comprising the aforementioned wound healing agent. The agent for improving, healing or preventing ligament damage or rupture according to the present invention has the effect of improving, promoting or preventing ligament damage or rupture.
The ligament improving or reinforcing agent according to the present invention is characterized by comprising the aforementioned wound healing agent. The ligament improving or reinforcing agent according to the present invention has an effect of improving or reinforcing the ligament.
本発明に係る競走馬の屈腱の炎症、損傷または断裂に対する改善、治癒促進または予防剤は、前述の創傷治癒剤から成ることを特徴とする。本発明に係る競走馬の屈腱の炎症、損傷または断裂に対する改善、治癒促進または予防剤は、競走馬の屈腱の炎症、損傷または断裂を改善、治癒促進または予防する効果を有する。
本発明に係る競走馬の屈腱の改善または強化剤は、前述の創傷治癒剤から成ることを特徴とする。本発明に係る競走馬の屈腱の改善または強化剤は、競走馬の屈腱を改善または強化する効果を有する。
The agent for improving, accelerating or preventing healing of inflammation, damage or tear of a flexor tendon of a racehorse according to the present invention is characterized by comprising the aforementioned wound healing agent. The agent for improving, healing or preventing the bending, tendon inflammation, damage or tear of the racehorse according to the present invention has the effect of improving, promoting or preventing the inflammation, damage or tear of the flexor tendon of the racehorse.
The agent for improving or reinforcing a flexor tendon of a racehorse according to the present invention comprises the aforementioned wound healing agent. The agent for improving or reinforcing the flexor tendon of the racehorse according to the present invention has the effect of improving or strengthening the flexor tendon of the racehorse.
本発明に係る競走馬の靱帯の損傷または断裂に対する改善、治癒促進または予防剤は、前述の創傷治癒剤から成ることを特徴とする。本発明に係る競走馬の靱帯の損傷または断裂に対する改善、治癒促進または予防剤は、競走馬の靱帯の損傷または断裂を改善、治癒促進または予防する効果を有する。
本発明に係る競走馬の靱帯の改善または強化剤は、前述の創傷治癒剤から成ることを特徴とする。本発明に係る競走馬の靱帯の改善または強化剤は、競走馬の靱帯を改善または強化する効果を有する。
The agent for improving, healing or preventing a ligament damage or rupture of a racehorse according to the present invention comprises the aforementioned wound healing agent. The improvement, healing promotion or prevention agent for damage or rupture of the ligament of the racehorse according to the present invention has an effect of improving, promoting or preventing the damage or rupture of the ligament of the racehorse.
The ligament improving or reinforcing agent for racehorses according to the present invention is characterized by comprising the aforementioned wound healing agent. The ligament improving or reinforcing agent for a racehorse according to the present invention has an effect of improving or strengthening the ligament of a racehorse.
すなわち、本発明に係る腱の損傷または断裂に対する改善、治癒促進または予防剤・本発明に係る腱の改善または強化剤・本発明に係る靱帯の損傷または断裂に対する改善、治癒促進または予防剤・本発明に係る靱帯の改善または強化剤・本発明に係る競走馬の屈腱の炎症、損傷または断裂に対する改善、治癒促進または予防剤・本発明に係る競走馬の屈腱の改善または強化剤は、前述の創傷治癒剤・本発明に係る競走馬の靱帯の損傷または断裂に対する改善、治癒促進または予防剤・本発明に係る競走馬の靱帯の改善または強化剤（以下、腱等改善剤という）は、コラーゲンまたはゼラチンをコラゲナーゼにより分解して得られる分解物であって、アミノ酸配列が（Ｇｌｙ−Ｘ−Ｙ）n（式中、Ｇｌｙはグリシン残基を表し、ＸおよびＹはグリシン以外の任意のアミノ酸残基を表し、ｎは正の整数を表す）で表されるペプチドを含むものを有効成分とする。好ましくは、腱等改善剤は、コラーゲンまたはゼラチンをコラゲナーゼにより分解して得られる分解物であって、アミノ酸配列が（Ｇｌｙ−Ｘ−Ｙ）n（式中、Ｇｌｙはグリシン残基を表し、ＸおよびＹはグリシン以外の任意のアミノ酸残基を表し、ｎは正の整数を表す）で表されるペプチドを有効成分とする。さらに好ましくは、腱等改善剤は、前記分解物はアミノ酸配列がＧｌｙ−Ｘ−Ｙで表されるトリペプチドを５質量％以上含む。また、腱等改善剤は、コラーゲンまたはゼラチンをコラゲナーゼにより分解して得られる分解物であって、アミノ酸配列がＧｌｙ−Ｘ−Ｙ（式中、Ｇｌｙはグリシン残基を表し、ＸおよびＹはグリシン以外の任意のアミノ酸残基を表す）で表されるトリペプチドを有効成分とすることが好ましい。また、腱等改善剤は、アスコルビン酸またはその誘導体が配合されていることが好ましい。 That is, an improvement, healing promotion or prevention agent for tendon damage or rupture according to the present invention. An improvement or strengthening agent for tendon according to the present invention. An improvement, healing promotion or prevention agent for ligament damage or rupture according to the present invention. Improvement or reinforcement agent for ligaments according to the invention- Improvement, healing promotion or prevention agent for inflammation, damage or tearing of the tendon of the racehorse according to the invention- Improvement or reinforcement agent for the tendon of the racehorse according to the invention is the aforementioned Wound healing agent ・ Improvement, healing promotion or prevention of damage or rupture of ligament of racehorse according to the present invention ・ Improvement or strengthening agent of racehorse ligament according to the present invention (hereinafter referred to as a tendon improving agent) is collagen Alternatively, a degradation product obtained by degrading gelatin with collagenase, the amino acid sequence of which is (Gly-XY) n (where Gly represents a glycine residue, and X and Y are An active ingredient includes a peptide represented by any amino acid residue other than lysine, and n represents a positive integer. Preferably, the tendon-improving agent is a degradation product obtained by degrading collagen or gelatin with collagenase, and the amino acid sequence is (Gly-XY) n (wherein Gly represents a glycine residue, X And Y represents any amino acid residue other than glycine, and n represents a positive integer). More preferably, in the agent for improving tendon and the like, the degradation product contains 5% by mass or more of a tripeptide whose amino acid sequence is represented by Gly-XY. An agent for improving tendons and the like is a degradation product obtained by degrading collagen or gelatin with collagenase, and has an amino acid sequence of Gly-XY (where Gly represents a glycine residue, and X and Y represent glycine. It is preferable to use a tripeptide represented by (representing any amino acid residue other than) as an active ingredient. The tendon and the like improving agent preferably contains ascorbic acid or a derivative thereof.
腱等改善剤は、ヒト、競走馬のほかの哺乳類、鳥類、爬虫類、魚類に用いられてもよい。対象となる腱は、アキレス腱のほか、肩、腕その他、部位を問わない。対象となる靱帯もまた、部位を問わない。対象となる競走馬の屈腱、靭帯は、脚の屈腱、靭帯であることが好ましい。 The tendon improving agent may be used for mammals other than humans, racehorses, birds, reptiles, and fish. The target tendons may be Achilles tendons, shoulders, arms, or other parts. The target ligament may be any part. The flexion tendon and ligament of the target racehorse are preferably the flexion tendon and ligament of the leg.
本発明に係る機能性食品は、前述の創傷治癒剤または腱等改善剤を０．００５質量％以上含むことを特徴とする。
本発明に係る医薬品は、前述の創傷治癒剤または腱等改善剤を０．００５質量％以上含むことを特徴とする。
The functional food according to the present invention contains 0.005% by mass or more of the above-described wound healing agent or tendon improving agent.
The pharmaceutical product according to the present invention is characterized by containing 0.005% by mass or more of the aforementioned wound healing agent or tendon improving agent.
本発明において、コラゲナーゼには、Clostridium histolyticum, Bacillusなどの細菌、Streptomyces parvulus などの放線菌、乳酸菌、酵母または真菌など由来で、コラーゲン特有のアミノ酸配列：（Gly-X-Y ）n のグリシンのアミノ基側を特異的に切断する酵素を用いる。また、コラゲナーゼは、これらの酵素遺伝子を遺伝子工学的に特定のベクターに組み込んで、乳酸菌や酵母などの他の菌体または動物に産生させて得られた遺伝子組み替えによる酵素で、類似の基質特異性を有するコラゲナーゼ様酵素であってもよい。 In the present invention, collagenase is derived from bacteria such as Clostridium histolyticum and Bacillus, actinomycetes such as Streptomyces parvulus, lactic acid bacteria, yeasts or fungi, and has a unique amino acid sequence of collagen: (Gly-XY) n on the amino group side of glycine An enzyme that specifically cleaves is used. Collagenase is a gene recombination enzyme obtained by incorporating these enzyme genes into specific vectors by genetic engineering and producing them in other cells or animals such as lactic acid bacteria and yeast, and has similar substrate specificity. It may be a collagenase-like enzyme having
コラゲナーゼは遊離の形で使用しても良いし、コラゲナーゼを物理吸着法または化学結合法によって各種の担体に結合させた固定化酵素として使用しても良い。また、コラゲナーゼによる酵素分解の方法には、(a) バッチ法、(b) カラム法または(c) これらを組み合わせた方法などがある。これら（ａ）〜（ｃ）の方法による製造ラインと、使用するコラゲナーゼの形態との組み合わせは、いろいろな方式を採用することが可能である。 Collagenase may be used in a free form, or may be used as an immobilized enzyme in which collagenase is bound to various carriers by a physical adsorption method or a chemical binding method. Examples of the enzymatic degradation method using collagenase include (a) a batch method, (b) a column method, and (c) a method combining these. Various combinations of the production line by the methods (a) to (c) and the form of collagenase to be used can be employed.
前記トリペプチドは、アミノ酸配列が、（Gly-Ala-Ｘ１）、Ｘ１はAla 、Arg 、Asp 、Hyp 、Lys 、Ser もしくはVal 、（Gly-Asp-Ｘ２）、Ｘ２はAla もしくはThr 、(Gly-Gln- Ｘ３）、Ｘ３は Glu、 HypもしくはSer 、（Gly-Pro-Ｘ４）、Ｘ４はAla 、Arg 、Hyp 、Hyl 、Gln 、Met 、Ile 、Lys 、Pro 、Ser 、Thr もしくはVal 、(Gly-Ser- Ｘ５）、Ｘ５は Ala、Glu もしくはHyp 、（Gly-Lys-Ｘ６）、Ｘ６はAsp 、Pro もしくはSer 、（Gly-Leu-Hyp ）、または(Gly-Val- Ｘ７）、Ｘ７は HypもしくはArg の一般式で示されるトリペプチドの１種または２種以上の混合物から成ることが好ましい。特に、前記トリペプチドは、アミノ酸配列が、（Gly-Ala-Arg ）、（Gly-Ala-Hyp ）、（Gly-Ala-Lys ）、（Gly-Pro-Ala ）、（Gly-Pro-Arg ）、（Gly-Pro-Hyp ）および(Gly-Pro-Ser)の一般式で示されるトリペプチドの混合物であることが好ましい。前記トリペプチドは、アミノ酸を用いた合成技術または発酵技術によって得られたものであってもよい。 The tripeptide has an amino acid sequence of (Gly-Ala-X1), X1 is Ala, Arg, Asp, Hyp, Lys, Ser or Val, (Gly-Asp-X2), X2 is Ala or Thr, (Gly- Gln-X3), X3 is Glu, Hyp or Ser, (Gly-Pro-X4), X4 is Ala, Arg, Hyp, Hyl, Gln, Met, Ile, Lys, Pro, Ser, Thr or Val, (Gly- Ser-X5), X5 is Ala, Glu or Hyp, (Gly-Lys-X6), X6 is Asp, Pro or Ser, (Gly-Leu-Hyp), or (Gly-Val-X7), X7 is Hyp or It is preferably composed of one or a mixture of two or more tripeptides represented by the general formula of Arg. In particular, the tripeptide has an amino acid sequence of (Gly-Ala-Arg), (Gly-Ala-Hyp), (Gly-Ala-Lys), (Gly-Pro-Ala), (Gly-Pro-Arg) , (Gly-Pro-Hyp) and (Gly-Pro-Ser) are preferred to be a mixture of tripeptides represented by the general formulas. The tripeptide may be obtained by a synthesis technique using amino acids or a fermentation technique.
本発明に係る創傷治癒剤、腱等改善剤、機能性食品および医薬品は、カルシウムおよびビタミンＤのいずれか一方または両方を含んでいてもよい。また、グリチルリチン酸エステルまたはその誘導体を含んでいてもよい。また、ヒアルロン酸を含んでいてもよい。本発明に係る創傷治癒剤、腱等改善剤、機能性食品および医薬品は、液剤、固形剤、カプセル剤、その他、形態を問わない。また、甘味剤、賦形剤、充填剤、増量剤、表面活性剤、着色剤、香料、防腐剤などその他の添加物を添加してあってもよい。本発明に係る創傷治癒剤、腱等改善剤および医薬品は、経口投与のほか、肛門投与、静脈投与、皮下投与、筋肉内投与などの方法で投与されてもよい。投与量は、投与対象の年齢、体重、症状などに応じて、適宜、変更することが好ましい。 The wound healing agent, tendon improving agent, functional food and pharmaceutical product according to the present invention may contain either or both of calcium and vitamin D. Further, it may contain glycyrrhizic acid ester or a derivative thereof. Moreover, hyaluronic acid may be included. The wound healing agent, tendon and other improving agents, functional foods and pharmaceuticals according to the present invention may be in the form of a liquid, a solid, a capsule, or the like. Moreover, you may add other additives, such as a sweetening agent, an excipient | filler, a filler, a bulking agent, a surface active agent, a coloring agent, a fragrance | flavor, and an antiseptic | preservative. In addition to oral administration, the wound healing agent, tendon-improving agent and pharmaceutical agent according to the present invention may be administered by methods such as anal administration, intravenous administration, subcutaneous administration, and intramuscular administration. It is preferable to appropriately change the dose according to the age, weight, symptoms, etc. of the administration subject.
本発明によれば、新規で有用な創傷治癒剤、腱の損傷または断裂に対する改善、治癒促進または予防剤、腱の改善または強化剤、靱帯の損傷または断裂に対する改善、治癒促進または予防剤、靱帯の改善または強化剤、競走馬の屈腱の炎症、損傷または断裂に対する改善、治癒促進または予防剤、競走馬の屈腱の改善または強化剤、競走馬の靱帯の損傷または断裂に対する改善、治癒促進または予防剤、競走馬の靱帯の改善または強化剤、その創傷治癒剤を含む機能性食品ならびに医薬品を提供することができる。 According to the present invention, a novel and useful wound healing agent, improvement against tendon damage or tear, healing promotion or prevention agent, tendon improvement or strengthening agent, improvement against ligament damage or tear, healing promotion or prevention agent, ligament Improvement or strengthening agent, racehorse flexor tendon inflammation, damage or rupture improvement, healing promotion or prevention agent, racehorse flexion tendon improvement or enhancement agent, racehorse ligament damage or rupture improvement, healing promotion or prevention An agent, a racehorse ligament improvement or strengthening agent, a functional food containing the wound healing agent, and a medicine can be provided.
本発明の実施の形態の創傷治癒剤は、コラーゲン成分あるいはゼラチン成分をコラゲナーゼ酵素を用いて特異的に分解して得られるペプチド組成物であって、アミノ酸配列が Ｇｌｙ−Ｘ−Ｙ のトリペプチド（以下コラーゲントリペプチド）を５％以上含有するものから成る。アミノ酸配列の Ｇｌｙ−Ｘ−Ｙ の式で、ＸおよびＹはグリシン（Ｇｌｙ）以外の任意のアミノ酸残基、例えば、プロリン、アラニン、あるいはヒドロキシプロリンなどである。 The wound healing agent according to the embodiment of the present invention is a peptide composition obtained by specifically degrading a collagen component or a gelatin component using a collagenase enzyme, and is a tripeptide (Gly-XY) having an amino acid sequence of Gly-XY The collagen tripeptide) contains 5% or more. In the amino acid sequence Gly-XY, X and Y are any amino acid residue other than glycine (Gly), such as proline, alanine, or hydroxyproline.
本発明の実施の形態の創傷治癒剤は、特開平7-82299号公報に記載のペプチド組成物の調整法、特開平9-176196号公報に記載の非抗原性安定化剤の調整法、または特開平11-12196号公報に記載の低抗原性安定化剤の調整法と同一の方法で調整することができ、概略的には、コラーゲン成分あるいはゼラチン成分をコラゲナーゼ酵素を用いて特異的に分解して得られる。 The wound healing agent of the embodiment of the present invention is a method for preparing a peptide composition described in JP-A-7-82299, a method for preparing a non-antigenic stabilizer described in JP-A-9-176196, or It can be prepared by the same method as the preparation method of the low antigenic stabilizer described in JP-A-11-12196. In general, the collagen component or the gelatin component is specifically decomposed using a collagenase enzyme. Is obtained.
本発明の実施の形態の創傷治癒剤は、ゼラチン成分またはコラーゲン成分を出発原料とし、酵素の回収率を良くするために固定化コラゲナーゼ酵素を用いたバッチ法やカラム法によるバイオリアクター方式で製造することができる。すなわち、コラゲナーゼ酵素を物理的吸着法あるいは化学結合法によって各種の担体、例えば、キトパールなどに結合させ、クロマトグラフィー用のカラムに充填し、一定温度下、可溶化させたゼラチン溶液あるいは分解が生じない程度の温度、望ましくは４０〜４５℃で変性させたコラーゲンをカラムに通して酵素分解をさせる。この際のコラーゲン原料の送液速度は、固定化酵素の活性および必要とされる分解度合いに応じて適宜選択される。 The wound healing agent according to the embodiment of the present invention is manufactured by a bioreactor method using a batch method or a column method using an immobilized collagenase enzyme in order to improve the enzyme recovery rate, using a gelatin component or a collagen component as a starting material. be able to. That is, collagenase enzyme is bound to various carriers such as chitopearl by physical adsorption method or chemical bonding method, and packed in a chromatography column, solubilized gelatin solution or decomposition does not occur at a constant temperature. Collagen denatured at a moderate temperature, preferably 40-45 ° C., is passed through the column for enzymatic degradation. In this case, the feeding rate of the collagen raw material is appropriately selected according to the activity of the immobilized enzyme and the required degree of degradation.
この調整法により得られた、アミノ酸配列が（Ｇｌｙ−Ｘ−Ｙ）nで表されるペプチドを含む分解物、好ましくはコラーゲントリペプチドを５％以上含有するもの、さらに好ましくはコラーゲントリペプチドは、創傷治癒剤、腱の損傷または断裂に対する改善、治癒促進または予防剤、腱の改善または強化剤、靱帯の損傷または断裂に対する改善、治癒促進または予防剤、靱帯の改善または強化剤、競走馬の屈腱の炎症、損傷または断裂に対する改善、治癒促進または予防剤、競走馬の屈腱の改善または強化剤、競走馬の靱帯の損傷または断裂に対する改善、治癒促進または予防剤、競走馬の靱帯の改善または強化剤として用いることができる。 A degradation product containing a peptide represented by the amino acid sequence (Gly-X—Y) n obtained by this adjustment method, preferably containing 5% or more of a collagen tripeptide, more preferably a collagen tripeptide, Wound healing agent, improvement against tendon damage or tear, healing promotion or prevention agent, tendon improvement or strengthening agent, improvement against ligament damage or tear, healing promotion or prevention agent, ligament improvement or strengthening agent, racehorse flexor tendon Inflammation, injury or tear improvement, healing promotion or prevention agent, racehorse flexor tendon improvement or strengthening agent, racehorse ligament injury or tear improvement, healing promotion or prevention agent, racehorse ligament improvement or enhancement It can be used as an agent.
本発明の実施の形態の創傷治癒剤は、コラーゲントリペプチドを５％以上含有する前記ペプチド組成物にアスコルビン酸もしくはその誘導体を配合することにより、さらに効果を増す。 The effect of the wound healing agent according to the embodiment of the present invention is further increased by blending ascorbic acid or a derivative thereof into the peptide composition containing 5% or more of collagen tripeptide.
本発明の実施の形態の創傷治癒剤は、もともと消化吸収されやすいタンパク質であることが知られているコラーゲンやゼラチン、およびその加水分解物と同等ないしはそれ以上に速やかに、且つ効率よく消化吸収される。 The wound healing agent according to the embodiment of the present invention is digested and absorbed as quickly and efficiently as collagen or gelatin, which is known to be a protein that is easily digested and absorbed, and hydrolysates thereof. The
本発明の実施の形態の機能性食品は、前記創傷治癒剤を０．００５質量％以上含むことを特徴とする。その機能性食品は、健康食品であっても、栄養機能食品や特定保健用食品などの保健機能食品であっても良く、コラーゲン産生促進、骨芽細胞または線維芽細胞の増殖促進、骨の強化または密度向上、生体組織の新陳代謝促進、皮膚賦活、皮膚または骨の加齢による機能低下の予防または改善、慢性リウマチ、変形関節症の改善および骨折治癒促進などの機能性に加え、腱断裂、靭帯損傷、屈腱炎などの治癒を促進する。 The functional food according to an embodiment of the present invention includes 0.005% by mass or more of the wound healing agent. The functional food may be a health food, a health functional food such as a nutritional functional food or a food for specified health use, promote collagen production, promote osteoblast or fibroblast proliferation, strengthen bone In addition to functionalities such as density enhancement, vital tissue metabolism promotion, skin activation, prevention or improvement of functional deterioration due to aging of skin or bone, chronic rheumatism, osteoarthritis improvement and fracture healing promotion, tendon rupture, ligament Promotes healing of injuries, tendonitis, etc.
本発明の実施の形態の医薬品は、前記創傷治癒剤を０．００５質量％以上含むことを特徴とする。その医薬品の範疇には、医薬部外品、化粧品も含み、コラーゲン産生促進、骨芽細胞または線維芽細胞の増殖促進、骨の強化または密度向上、生体組織の新陳代謝促進、皮膚賦活、皮膚または骨の加齢による機能低下の予防または改善、慢性リウマチ、変形関節症の改善および骨折治癒促進などの機能性に加え、腱断裂、靭帯損傷、屈腱炎などの治癒を促進する機能性を有する。 The pharmaceutical of the embodiment of the present invention is characterized by containing 0.005% by mass or more of the wound healing agent. The category of pharmaceuticals includes quasi-drugs and cosmetics, promotes collagen production, promotes proliferation of osteoblasts or fibroblasts, strengthens or increases bone density, promotes metabolism of biological tissues, activates skin, skin or bone In addition to functionalities such as prevention or improvement of functional deterioration due to aging, chronic rheumatism, improvement of osteoarthritis and promotion of fracture healing, it has the ability to promote healing of tendon rupture, ligament injury, flexor tendonitis and the like.
本発明の実施の形態の創傷治癒剤の出発原料であるコラーゲンまたはゼラチンには、牛、豚、鳥、鯨その他の鳥獣類などの骨、軟骨、皮、腱、または鮭、鮫その他の魚類などの魚皮、ウロコなどを原料として調製されたものを用いることができ、そのタイプは問わない。コラゲナーゼには、コラーゲン特有のアミノ酸配列：（Gly-X-Y）n のグリシンのアミノ基側を特異的に切断する酵素を用いる。 Collagen or gelatin, which is a starting material of the wound healing agent according to the embodiment of the present invention, includes bone, cartilage, skin, tendon, or shark, salmon, other fish, etc. such as cattle, pigs, birds, whales and other birds and beasts. What was prepared using fish skin, scales, etc. as a raw material can be used. Collagenase uses an enzyme that specifically cleaves the amino group side of glycine in the amino acid sequence unique to collagen: (Gly-X-Y) n.
以下、実施例により本発明を詳細に説明するが、この実施例は、本発明の範囲を何ら制限するものではない。また、本発明に係る創傷治癒剤において、コラーゲントリペプチドまたはコラーゲントリペプチドを５質量％以上含むコラーゲンペプチドを機能性食品、医薬品に配合する方法は、既に公知となっている方法に準じて実施することができる。 EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this Example does not restrict | limit the scope of the present invention at all. Further, in the wound healing agent according to the present invention, a method of blending collagen tripeptide or collagen peptide containing 5% by mass or more of collagen tripeptide into functional foods and pharmaceuticals is performed in accordance with a method already known. be able to.
以下の方法により、創傷治癒剤を製造した。
高純度ゼラチン（ゼライス社製） 50 gを 1,000 mlの 20 mM Tris-HCl 緩衝液（pH 7.4）／0.1M NaCl に加温しながら溶解後、50℃に冷却した。 酵素分解用の固定化酵素は、100 mg のコラゲナーゼ酵素（ワシントン社製、type IVから精製した）を 50 g の粒子担体に結合させて調製した。担体への結合量は結合前後の 280 nm における吸光度の変化を計測して算出したが、９９％以上の結合率であった。使用時、本固定化酵素をカラム式バイオリアクターに充填し、20 mM Tris-HCl 緩衝液（pH 7.4）／0.1M NaCl 緩衝液で良く洗浄・平衡化を行った。
A wound healing agent was produced by the following method.
50 g of high-purity gelatin (manufactured by Zerais) was dissolved in 1,000 ml of 20 mM Tris-HCl buffer (pH 7.4) /0.1 M NaCl while heating, and then cooled to 50 ° C. The immobilized enzyme for enzymatic degradation was prepared by binding 100 mg of collagenase enzyme (purified from type IV, manufactured by Washington) to 50 g of particle carrier. The amount bound to the carrier was calculated by measuring the change in absorbance at 280 nm before and after the binding, and the binding rate was 99% or more. At the time of use, this immobilized enzyme was packed in a column type bioreactor and washed and equilibrated well with 20 mM Tris-HCl buffer (pH 7.4) /0.1 M NaCl buffer.
原料であるコラーゲンあるいはゼラチンを、上記工程で調製されたコラゲナーゼ酵素固定化カラムにアプライし、カラム法による酵素分解を行った。この間、流速は毎分 20 〜 60 ml に、また、カラムの温度は 39 ± 1 ℃にコントロールした。カラムから出てきた酵素反応終了液を分取し、0.45 μm のフィルターで濾過を行った。この濾液をスプレードライ（噴霧乾燥器）で粉末化した。 Collagen or gelatin as a raw material was applied to the collagenase enzyme-immobilized column prepared in the above step, and enzymatic degradation was performed by the column method. During this time, the flow rate was controlled at 20-60 ml / min, and the column temperature was controlled at 39 ± 1 ° C. The enzyme reaction finished solution that came out of the column was collected and filtered through a 0.45 μm filter. The filtrate was pulverized by spray drying (spray dryer).
得られたコラーゲントリペプチド含量が５質量％以上（本実施例では約１５質量％）のコラーゲンペプチドを凍結乾燥した後、Ｎ末端アミノ酸についてエドマン分解法によって検定した。その結果、Ｎ末端側のアミノ酸は 97.5％がグリシンであることが判明し、本発明によるところのコラーゲンに特徴的なアミノ酸配列である (Gly-X-Y)n 構造を保持していることが証明された。本実施例の創傷治癒剤は、こうして製造されたコラーゲンペプチドから成る。 The obtained collagen peptide having a collagen tripeptide content of 5% by mass or more (about 15% by mass in this example) was freeze-dried, and then the N-terminal amino acid was assayed by the Edman degradation method. As a result, 97.5% of the amino acids on the N-terminal side were found to be glycine, and it was proved to retain the (Gly-XY) n structure, which is an amino acid sequence characteristic of collagen according to the present invention. It was. The wound healing agent of this example is composed of the collagen peptide thus produced.
＜体内動態試験＞
実施例１により得られた創傷治癒剤：コラーゲントリペプチドを放射性同位元素：トリチウム（３Ｈ）で標識したもの（以下、３Ｈ-コラーゲントリペプチド）、あるいは比較対照として３Ｈ-プロリンを、ラットに経口投与し、（１）血漿中放射能濃度の測定、（２）全身オートラジオグラフィーにより、被験物質のラット体内における吸収及び分布について検討した。
<Pharmacokinetic study>
Oral administration of the wound healing agent obtained in Example 1: collagen tripeptide labeled with radioisotope: tritium (3H) (hereinafter 3H-collagen tripeptide) or 3H-proline as a comparative control to rats. Then, the absorption and distribution of the test substance in the rat body were examined by (1) measurement of radioactivity concentration in plasma and (2) whole body autoradiography.
（１）血漿中放射能濃度の測定
血漿中放射能濃度の測定の結果、３Ｈ-コラーゲントリペプチドを経口投与から 5分後において、既に血漿中放射能濃度は 1.4μg eq./mL を示し、その後、経時的に上昇し、最大濃度：42.7μg eq./mL に達した。次いで、徐々に低下する様子が観察され、対照として用いたアミノ酸：３Ｈ-プロリンよりも、吸収速度が速いことが確認された。
(1) Measurement of plasma radioactivity concentration As a result of measurement of plasma radioactivity concentration, plasma radioactivity concentration was already 1.4 μg eq./mL 5 minutes after oral administration of 3H-collagen tripeptide, Thereafter, it increased with time and reached the maximum concentration: 42.7 μg eq./mL. Subsequently, a gradual decrease was observed, and it was confirmed that the absorption rate was faster than that of the amino acid used as a control: 3H-proline.
（２）全身オートラジオグラフィー
３Ｈ-コラーゲントリペプチドを投与したラットをエーテル麻酔死させた後、全身オートラジオグラムを作成し、各種組織への放射能分布および経時変化について検討した結果、１回投与２４時間後の全身オートラジオグラムにより、腱、靭帯、骨髄、骨膜、胸腺、消化管、粘膜、皮膚などへの分布が確認された。投与回数５回、１２回の全身オートラジオグラムでは、さらに、分布密度が濃く観察され、各組織への分布が明らかとなった。その全身オートラジオグラフィーを図１に示す。
(2) Whole body autoradiography After the rats administered with 3H-collagen tripeptide were killed by ether anesthesia, whole body autoradiogram was prepared, and the radioactivity distribution to various tissues and changes with time were examined. Whole body autoradiogram after 24 hours confirmed distribution to tendons, ligaments, bone marrow, periosteum, thymus, gastrointestinal tract, mucous membrane, skin and the like. In the whole body autoradiogram with 5 and 12 administrations, the distribution density was further observed, and the distribution to each tissue became clear. The whole body autoradiography is shown in FIG.
＜コラーゲン産生促進作用＞
５％ ＦＢＳ−Ｅａｇｌｅ ＭＥＭ培地に、０μＭ あるいは １μＭ のアスコルビン酸（ＡＳＡ）、および ０μｇ／ｍＬ あるいは ３．０μｇ／ｍＬ 濃度の実施例１により得られた創傷治癒剤：コラーゲントリペプチドを培養液に添加し、その培養液でヒト正常線維芽細胞を３日間培養した。培養終了後それぞれの培養上清を採取し、各培養上清中に分泌された typeＩ コラーゲンをウサギ抗ヒト typeＩ コラーゲン抗血清を用いたＥＬＩＳＡ法により測定した。その結果、図２に示すように、a.実施例１により得られた創傷治癒剤の単独添加により、コラーゲン産生能が上昇する、b.１μＭ ＡＳＡ単独添加によりコラーゲン産生が誘導される、c.実施例１により得られた創傷治癒剤は、ＡＳＡ添加によって誘導されるコラーゲン産生をさらに増強する、ことが明らかとなった。
<Promoting collagen production>
To 5% FBS-Eagle MEM medium, 0 μM or 1 μM ascorbic acid (ASA), and the wound healing agent obtained in Example 1 at a concentration of 0 μg / mL or 3.0 μg / mL: collagen tripeptide added to the culture medium Then, normal human fibroblasts were cultured for 3 days in the culture solution. After completion of the culture, each culture supernatant was collected, and type I collagen secreted in each culture supernatant was measured by ELISA using rabbit anti-human type I collagen antiserum. As a result, as shown in FIG. 2, a. The ability to produce collagen is increased by adding the wound healing agent obtained in Example 1 alone, b. Collagen production is induced by adding only 1 μM ASA, c. It was revealed that the wound healing agent obtained in Example 1 further enhances collagen production induced by addition of ASA.
＜骨折治癒促進作用＞
７週齢のＳＤ系雄ラットの左大腿骨に人為的に骨折を施して骨折モデルラットを作製し、モデル作製翌日から１２週間、１日１回、実施例１により得られた創傷治癒剤：コラーゲントリペプチドをゾンデで、０，８０，５００ｍｇ/ｋｇ経口投与した。投与終了翌日、大腿骨（左右）を摘出し、軟Ｘ線写真撮影（ソフテックス株式会社製ＳＯＦＴＥＸ-Ｍ-６０使用）及び骨強度測定（マルト−株式会社製ＭＺ５００Ｄ使用）を実施し、骨折の治癒状態を判定した。
<Fracture healing promotion effect>
A fracture model rat was prepared by artificially fractured the left femur of a 7-week-old SD male rat. The wound healing agent obtained in Example 1 once a day for 12 weeks from the day after the model was prepared: Collagen tripeptide was orally administered with a sonde at 0,80,500 mg / kg. The day after the administration was completed, the femur (left and right) was removed, soft X-ray photography (using Softex-M-60, Softex-M-60) and bone strength measurement (using Malto-MZ500D) were performed. The healing state was determined.
（１）軟Ｘ線写真
軟Ｘ線写真（図３）から、実施例１により得られた創傷治癒剤：コラーゲントリペプチドを投与した群（８０,５００ｍｇ/ｋｇ）では、対照群（０ｍｇ/ｋｇ）よりも皮質骨の連続性が増し、骨折線が薄くなる傾向が認められ、骨折治癒の亢進傾向が確認された。
(1) Soft X-ray photograph From the soft X-ray photograph (FIG. 3), the wound healing agent obtained by Example 1: collagen tripeptide administered group (80,500 mg / kg), the control group (0 mg / kg) ), The continuity of cortical bone increased, the fracture line tended to be thinner, and a tendency to promote fracture healing was confirmed.
（２）骨強度測定
左右大腿骨の骨幹部で三点曲げ試験（支点間距離：１８ｍｍ、前面を上方、近位側を右側とし２０ｍｍ/６０ｓで破断）を行ったところ、ａ．破断時間、ｂ．破断変位、ｃ．破断荷重、ｄ．最大荷重、e．堅さ、のいずれのパラメータについても、対照群（０ｍｇ/ｋｇ）では、骨折処置を施した左大腿骨と骨折処置を施していない右大腿骨の測定値間で大きな差が認められたのに対し、実施例１により得られた創傷治癒剤：コラーゲントリペプチドを投与した群（８０,５００ｍｇ/ｋｇ）では、差が小さく、健常な骨（右大腿骨）と同程度まで回復していることが確認された（図４乃至図８参照）。
(2) Measurement of bone strength When a three-point bending test (distance between fulcrums: 18 mm, front surface is upward, proximal side is right side and fractured at 20 mm / 60 s) was performed on the left and right femur shafts, a. Breaking time, b. Breaking displacement, c. Breaking load, d. Maximum load, e. For both parameters of stiffness, a large difference was observed in the control group (0 mg / kg) between the measurements of the left femur with and without the fracture treatment. In contrast, in the group (80,500 mg / kg) administered with the wound healing agent: collagen tripeptide obtained in Example 1, the difference was small, and it was restored to the same level as that of healthy bone (right femur). Was confirmed (see FIGS. 4 to 8).
＜アキレス腱断裂時における治癒促進作用＞
７週齢のＳＤ系雄ラットの左アキレス腱を人為的に切断してアキレス腱切断モデルラットを作製し、モデル作製直後から、１日１回２、３週間および４週間、実施例１により得られた創傷治癒剤：コラーゲントリペプチドをゾンデで、０，８０ｍｇ/ｋｇ経口投与した。投与終了日の翌日、アキレス腱周囲の腱を取り出し、組織観察および引っ張り試験を実施してアキレス腱の修復程度を判定した。
<Healing promotion effect when Achilles tendon is torn>
The 7-week-old SD male rat left Achilles tendon was artificially cut to produce an Achilles tendon cut model rat, which was obtained according to Example 1 once a day for 2, 3 and 4 weeks. Wound healing agent: Collagen tripeptide was orally administered at 0.80 mg / kg with a sonde. On the next day after the administration end, the tendon around the Achilles tendon was taken out, and a tissue observation and a tensile test were performed to determine the degree of repair of the Achilles tendon.
（１）組織観察
２週間目において、実施例１により得られた創傷治癒剤：コラーゲントリペプチドを投与した群（８０ｍｇ/ｋｇ）では、対照群（０ｍｇ/ｋｇ）よりも切断部位付近の炎症が抑制されており、血管の増生、弾性線維量の増加傾向が認められた。さらに、腫骨の破骨細胞数も少なく、骨の形状および腱と骨との結合部位の状態も正常側（右）に近づいており、修復の亢進傾向が認められた。４週目では、対照群においても大部分が修復している様子ではあったが、コラーゲントリペプチド投与群の方が、より正常状態に近かった。
(1) Tissue observation In the second week, the wound healing agent obtained in Example 1: collagen tripeptide administered group (80 mg / kg) showed more inflammation near the cut site than the control group (0 mg / kg). Suppression was observed, and vascular proliferation and a tendency to increase the amount of elastic fibers were observed. Furthermore, the number of osteoclasts in the tumor was small, and the shape of the bone and the state of the joint site between the tendon and the bone were close to the normal side (right), indicating a tendency for increased repair. At 4 weeks, most of the control group seemed to be restored, but the collagen tripeptide administration group was closer to normal.
（２）引っ張り試験
腱の部分と筋肉部分に縫合糸を巻き、巻いた糸の部分を圧縮し保定し、さらに腫骨の部分を押さえて引っ張り試験（５０ｍｍ/ｍｉｎ）を行ったところ、２週目において、破断荷重、最大荷重ともコラーゲントリペプチド投与群（８０ｍｇ/ｋｇ）の値が高く、より正常側（右）に近い値を示し、修復の亢進傾向が認められた（図９）。
(2) Tensile test A suture was wound around the tendon part and the muscle part, and the wound thread part was compressed and held, and further, a tensile test (50 mm / min) was carried out by pressing the tumor part. In the eyes, both the breaking load and the maximum load were high in the collagen tripeptide administration group (80 mg / kg), showing values closer to the normal side (right), and a tendency to promote repair was observed (FIG. 9).
Claims (15)
A pharmaceutical comprising 0.005% by mass or more of the wound healing agent according to any one of claims 1 to 5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2004097205A JP2005281186A (en) | 2004-03-29 | 2004-03-29 | Wound healing agent, agent for amelioration, promotion of healing or prevention of injure or the like of tendon or ligament, functional food and medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2004097205A JP2005281186A (en) | 2004-03-29 | 2004-03-29 | Wound healing agent, agent for amelioration, promotion of healing or prevention of injure or the like of tendon or ligament, functional food and medicine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2005281186A true JP2005281186A (en) | 2005-10-13 |
Family
ID=35179964
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2004097205A Pending JP2005281186A (en) | 2004-03-29 | 2004-03-29 | Wound healing agent, agent for amelioration, promotion of healing or prevention of injure or the like of tendon or ligament, functional food and medicine |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2005281186A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008231065A (en) * | 2007-03-23 | 2008-10-02 | Chisso Corp | Bone metabolism improver |
| JP2009035491A (en) * | 2007-07-31 | 2009-02-19 | Morinaga Milk Ind Co Ltd | Bedsore ameliorating agent |
| WO2010103837A1 (en) | 2009-03-11 | 2010-09-16 | ゼライス株式会社 | Progression inhibitor or prophylactic agent for atherosclerosis, blood cholesterol level-lowering agent, and functional food or food for specified health uses |
| JP2017088561A (en) * | 2015-11-13 | 2017-05-25 | ニュートリー株式会社 | Nutritive composition for promoting recovery of postoperative wound site and/or anastomotic site |
| CN107854716A (en) * | 2017-09-27 | 2018-03-30 | 广州润虹医药科技股份有限公司 | A kind of antibacterial dehumidification type dressing patch and preparation method thereof |
-
2004
- 2004-03-29 JP JP2004097205A patent/JP2005281186A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008231065A (en) * | 2007-03-23 | 2008-10-02 | Chisso Corp | Bone metabolism improver |
| JP2009035491A (en) * | 2007-07-31 | 2009-02-19 | Morinaga Milk Ind Co Ltd | Bedsore ameliorating agent |
| WO2010103837A1 (en) | 2009-03-11 | 2010-09-16 | ゼライス株式会社 | Progression inhibitor or prophylactic agent for atherosclerosis, blood cholesterol level-lowering agent, and functional food or food for specified health uses |
| US9339524B2 (en) | 2009-03-11 | 2016-05-17 | Jellice Co., Ltd. | Drug inhibiting the progression of atherosclerosis, preventive drug, blood cholesterol-lowering drug, functional food, and specific health food |
| JP2017088561A (en) * | 2015-11-13 | 2017-05-25 | ニュートリー株式会社 | Nutritive composition for promoting recovery of postoperative wound site and/or anastomotic site |
| CN107854716A (en) * | 2017-09-27 | 2018-03-30 | 广州润虹医药科技股份有限公司 | A kind of antibacterial dehumidification type dressing patch and preparation method thereof |
| CN107854716B (en) * | 2017-09-27 | 2020-10-02 | 广州润虹医药科技股份有限公司 | Antibacterial and dehumidifying dressing patch and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Nimni | Collagen | |
| Raman et al. | Fish collagen and its applications in food and pharmaceutical industry: A review | |
| DE69727332T2 (en) | Preparations containing collagen, vitamin D3 and calcium to strengthen the bone | |
| JP4947475B2 (en) | Hydrolyzate of avian cartilage and its preparation method and use | |
| EP2478923B1 (en) | A bone regeneration agent comprising gelatin | |
| JP5276813B2 (en) | Elastin-degrading peptide, method for producing elastin and its enzyme-degrading peptide | |
| RU2671401C2 (en) | Collagen hydrolysate and use thereof | |
| JP2003137807A (en) | Collagen-producing promoter, cosmetic, food and pharmaceutical containing the same and external preparation for preventing or improving dermatosis | |
| EP2428215B1 (en) | Recombinant gelatin for use in the treatment of ischemic and heart disease and others | |
| JP2002255847A (en) | Collagen production promoter, functional food and pharmaceutical preparation | |
| WO2010147109A1 (en) | Angiogenesis inducer comprising genetically modified gelatin and basic fibroblast growth factor | |
| KR100828494B1 (en) | A composition of a cosmetic essense and the method of prepating it for improvenent of wrinkle | |
| JP5459826B2 (en) | Elastin production promoter and vascular endothelial cell growth promoter in dermal fibroblasts | |
| JP3802721B2 (en) | Biological collagen synthesis promoter | |
| JP2805194B2 (en) | Peptide for increasing blood triglyceride concentration and blood triglyceride concentration increase inhibitor containing the peptide as an active ingredient | |
| WO2022067445A1 (en) | Bioactive collagen peptides, method of production thereof, and use thereof | |
| CN107412861B (en) | Bone repair gel of recombinant collagen compounded with chondroitin sulfate and polyethylene glycol | |
| Dybka et al. | Collagen hydrolysates as a new diet supplement | |
| JP2005281186A (en) | Wound healing agent, agent for amelioration, promotion of healing or prevention of injure or the like of tendon or ligament, functional food and medicine | |
| JP5778692B2 (en) | Disease inhibitor | |
| JP2006056904A (en) | Accelerator for synthesizing biogenic collagen | |
| JP7343864B2 (en) | Elastin production promoter in fibroblasts or chondrocytes | |
| JP6143218B2 (en) | Tendon and ligament function improving agent and pharmaceutical composition, food and feed containing the same | |
| Musayeva et al. | A review on collagen as a food supplement | |
| JP2004244369A (en) | Peptide composition, method for producing the same and peptide-added material |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20050902 |