JPH10194986A - Accelerator for amelioration and regeneration of transplanted hepatic function - Google Patents

Accelerator for amelioration and regeneration of transplanted hepatic function

Info

Publication number
JPH10194986A
JPH10194986A JP9168177A JP16817797A JPH10194986A JP H10194986 A JPH10194986 A JP H10194986A JP 9168177 A JP9168177 A JP 9168177A JP 16817797 A JP16817797 A JP 16817797A JP H10194986 A JPH10194986 A JP H10194986A
Authority
JP
Japan
Prior art keywords
tcf
liver
ii
regeneration
immunosuppressant
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
Application number
JP9168177A
Other languages
Japanese (ja)
Inventor
Keizo Sugimachi
Hideaki Uchiyama
秀昭 内山
圭蔵 杉町
Original Assignee
Snow Brand Milk Prod Co Ltd
雪印乳業株式会社
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
Priority to JP17055596 priority Critical
Priority to JP8-170555 priority
Priority to JP31553296 priority
Priority to JP8-315532 priority
Application filed by Snow Brand Milk Prod Co Ltd, 雪印乳業株式会社 filed Critical Snow Brand Milk Prod Co Ltd
Priority to JP9168177A priority patent/JPH10194986A/en
Publication of JPH10194986A publication Critical patent/JPH10194986A/en
Application status is Pending legal-status Critical

Links

Abstract

PROBLEM TO BE SOLVED: To obtain the subject medicine which accelerates the functional amelioration of transplanted hepatic tissues and its regeneration and can recover the hepatic function in the early stage after operation of liver transplantation by using a hepatocyte proliferation factor, particularly by combining the hepatocyte proliferation factor with an immunosuppressant in combination.
SOLUTION: This medicine contains a hepatocyte proliferation factor or both of the hepatocyte proliferation factor and an immunosuppressant. As the hepatocyte proliferation factor, TCF-11 is preferable, particularly when the TCF-11 and the immunosuppressant are used in combination, the activity is remarkably enhanced. The TCF-11 is a well-known protein desired from human fibroblast and is obtained by the method of purifying it after concentrating the culture medium of human fibroblast and allowing it to absorb into an ion exchanger or by means of genetic engineering. The TCF-11 can be parenterally administrated as an injection and the preparation containing 0.6-600mg/day/adult, preferably 6-60mg/day/adult, as the purified TCF-11 is administrated to patients before liver-transplanting. When the TCF-11 is used in combination with the immunosuppressant (e.g. tacrolimus), 0.1-100mg/day/adult is administrated.
COPYRIGHT: (C)1998,JPO

Description

【発明の詳細な説明】 DETAILED DESCRIPTION OF THE INVENTION

【0001】 [0001]

【発明の属する技術分野】本発明は、肝細胞増殖因子を有効成分とする移植肝機能改善・再生促進剤に関する。 The present invention relates to relates to transplant liver function improving and regeneration promoter having for an active ingredient to hepatocyte growth factor.
さらに、本発明は、肝細胞増殖因子と免疫抑制剤とを併用する移植肝機能改善・再生促進剤に関する。 Furthermore, the invention relates to transplant liver function improving and regeneration-promoting agent used in combination with hepatocyte growth factor and an immunosuppressive agent. 本発明により、移植した肝組織の機能改善及び再生が促進され、 The present invention is promoted improvements and regeneration of the transplanted liver tissue,
肝移植手術後の肝機能を早期に回復することができる。 It can be restored at an early stage liver function after surgery liver transplantation.

【0002】 [0002]

【従来の技術】現在、肝移植は末期肝硬変患者に対する唯一の根本的治療手段である。 At present, liver transplantation is the only fundamental treatment means for end-stage liver cirrhosis patients. しかし、脳死肝臓移植においては、深刻なドナー不足の問題がある。 However, in the brain death liver transplantation, there is a serious shortage of donors of the problem. この問題に対処する一つの方法として、ドナー肝を二つに分け、二人のレシピエントに移植する方法がある(分割肝移植)。 One way to address this problem, split the donor liver in two, there is a method of transplanting to two recipients (split liver transplantation). 当然ながら、レシピエントには理想肝容積よりも小さな移植片が移植されることになる。 Of course, so that the small graft is implanted than the ideal liver volume in the recipient. 小さな移植片は肝の予備能が劣っており、ラットでは理想肝容積の30% Small grafts are poor reserve of the liver, 30% of the ideal liver volume in rats
が限界とされている。 There has been a limit. また日本では、脳死肝移植は未だ社会的合意が得られておらず、当面は生体部分肝移植が主に行われていくことが予想される。 Also in Japan, brain death liver transplantation is not yet social consensus is obtained, the time being, it is expected that the biological part liver transplantation is going to be mainly carried out. この生体部分肝移植では、理想肝容積よりも小さな移植片が移植されるケースが多く、ことに成人の生体部分肝移植では当然のことながら、極めて小さな移植片が移植されることになる。 In the living body portion liver transplantation, many cases of small implant is implanted than the ideal liver volume, it will be appreciated by deciding on biological moiety liver transplantation in adults, would be extremely small graft is implanted. 脳死ドナーからの分割肝移植及び生体部分肝移植における、移植肝の機能をより早期に改善することが可能となれば、小さな部分肝の移植の施行が可能になると考えられる。 In the divided liver transplantation and biological part liver transplantation from brain-dead donors, if it is possible to improve the function of the transplanted liver earlier believed to allow the enforcement of the implantation of a small portion liver. しかし、このような部分移植肝の機能を改善する治療法に関しては、今までのところ、実際に臨床応用の可能性が期待できる報告はなかった。 However, with regard to the treatment method to improve the function of such partial transplant liver, so far, there were no reports that actually can be expected is a possibility of clinical applications.

【0003】 [0003]

【発明が解決しようとする課題】本発明者らは上述の状況に鑑み、肝臓移植後の移植肝機能を改善、また再生を促進させる物質を求め鋭意探索の結果、肝細胞増殖因子、特にTCF−IIが、その効果を有することを見出した。 In view of the present invention have found that the above conditions [0004], improve transplant liver function after liver transplantation, also search intensively sought substances which promote regeneration results, hepatocyte growth factor, in particular TCF -II was found to have the effect. さらに、肝細胞増殖因子、特にTCF−IIと免疫抑制剤とを併用することにより、その活性を著しく増強することを見出した。 Moreover, hepatocyte growth factor, in particular by combined use of TCF-II and immunosuppressant was found to significantly enhance its activity. 即ち、本発明は、肝細胞増殖因子、 That is, the present invention is, hepatocyte growth factor,
特にTCF−IIを有効成分とする移植肝機能改善・再生促進剤、及び肝細胞増殖因子、特にTCF−IIと免疫抑制剤とを併用する移植肝機能改善・再生促進剤を提供することを課題とする。 In particular object of the invention is to provide a TCF-II with an active ingredient transplanted liver function improving and regeneration promoter, and hepatocyte growth factor, in particular TCF-II and in combination with immunosuppressive drugs transplanted liver function improving and regeneration promoter to.

【0004】 [0004]

【課題を解決するための手段】本発明は、肝細胞増殖因子を有効成分とする移植肝機能改善・再生促進剤に関する。 SUMMARY OF THE INVENTION The present invention relates to transplant liver function improving and regeneration promoter having for an active ingredient to hepatocyte growth factor. さらに、本発明は肝細胞増殖因子と免疫抑制剤とを併用する移植肝機能改善・再生促進剤に関する。 Furthermore, the present invention relates to transplant liver function improving and regeneration-promoting agent used in combination with hepatocyte growth factor and an immunosuppressive agent. 本発明における肝細胞増殖因子としては、特にTCF−IIが好ましい。 The hepatocyte growth factor in the present invention, especially TCF-II are preferred. 本発明により、移植した肝組織の機能改善及び再生が促進され、肝移植手術後の肝機能を早期に回復することができる。 The present invention is promoted functional improvement and regeneration of the transplanted liver tissue can recover quickly liver function after surgery liver transplantation.

【0005】 [0005]

【発明の実施の形態】本発明の有効成分である肝細胞増殖因子は特に限定されず、特に好ましくはTCF−IIが用いられる。 Hepatocyte growth factor as an active ingredient of the present invention DETAILED DESCRIPTION OF THE INVENTION is not particularly limited, particularly preferably TCF-II is used. 本発明で用いられるTCF−IIは、ヒト線維芽細胞由来の公知の蛋白質であり、ヒト線維芽細胞由来のものは下記の特性を有する。 TCF-II for use in the present invention is a known protein derived from human fibroblasts, derived from human fibroblast cells have the following characteristics.

【0006】i) 分子量(SDS電気泳動法) 非還元下 : 78,000±2,000 又は74,000±2,000 還元下 : 52,000±2,000 (共通バンドA) 30,000±2,000 (バンドB) 26,000±2,000 (バンドC) ii) 等電点 : 7.4 〜 8.6 [0006] i) Molecular weight (SDS electrophoresis) under non-reducing: 78,000 ± 2,000 or 74,000 ± 2,000 reduction under: 52,000 ± 2,000 (common band A) 30,000 ± 2,000 (band B) 26,000 ± 2,000 (band C) ii) isoelectric point: 7.4 to 8.6

【0007】上記TCF−IIは、ヒト線維芽細胞培養液を濃縮しイオン交換体に吸着させ、その溶出液をアフィニティークロマトグラフィーを用いて精製する方法(WO [0007] The above TCF-II can be adsorbed on the ion exchanger was concentrated human fibroblast cultures, a method of the eluate is purified using affinity chromatography (WO
90/10651号公報)や、或いは遺伝子工学的手法 (WO92/0 90/10651 JP) and, or a genetic engineering technique (WO92 / 0
1053号公報)によって得ることができる。 Can be obtained by 1053 JP). この時、宿主細胞又は微生物の違いによる糖鎖の異なったものや、糖鎖の結合していないものであっても使用可能である。 In this case, different objects or the sugar chain by the difference of host cell or microorganism, be those unbound sugar chain can also be used. しかし、糖鎖は生体内の代謝速度に関係しているため、糖鎖の結合しているものを用いることが望ましい。 However, sugar chains because it relates to metabolic rate in vivo, it is desirable to use a bound sugar chain. これらの方法により得られたTCF−IIは、通常の単離精製法によってさらに濃縮、精製することができる。 TCF-II obtained by these methods can be further concentrated and purified by conventional isolation and purification methods. 例えば、 For example,
有機溶媒による沈殿法、塩析、ゲル濾過、モノクローナル抗体を用いたアフィニティークロマト、電気泳動法等が挙げられる。 Precipitation with organic solvents, salting out, gel filtration, affinity chromatography using monoclonal antibody, electrophoresis, or the like. モノクローナル抗体を用いたアフィニティークロマトによる精製は、特開平5−97号公報に開示されているモノクローナル抗体を用いて精製することができる。 Purification by affinity chromatography using monoclonal antibody can be purified using monoclonal antibodies disclosed in JP-A-5-103. 得られた精製TCF−IIは、凍結乾燥或いは凍結保存することができる。 Purification TCF-II obtained can be stored lyophilized or frozen. その他、TCF−IIと同様の活性を有するものであれば本発明と同様の薬剤として利用可能である。 Other available as similar agents as the present invention as long as it has the same activity as TCF-II. 例えば、TCF−II蛋白質と5アミノ酸の違いを有する蛋白質である肝細胞増殖因子(HGF; For example, hepatocyte growth factor (HGF is a protein having a difference of TCF-II protein and 5 amino acids;
特開昭63-22526号)、あるいは精製ScatterFactor(S JP 63-22526), ​​or purified ScatterFactor (S
F;Gherardi and Stocker, Nature, 346, 228(1990)) F; Gherardi and Stocker, Nature, 346, 228 (1990))
などが挙げられる。 And the like.

【0008】本発明の移植肝機能改善・再生促進剤は、 [0008] transplanted liver function improvement and regeneration-promoting agent of the present invention,
注射剤として静脈、筋肉内、及び皮下に非経口的に投与することができる。 Intravenously as an injection, intramuscular, and it may be administered parenterally subcutaneously. これらの製剤は公知の製剤学的製法に準じ製造され、必要に応じpH調整剤、緩衝剤、安定化剤等を添加することができる。 These formulations are prepared according to known pharmaceutical preparation, pH adjusting agents if necessary, buffering agents, can be added a stabilizer, and the like. 本発明の製剤は、肝移植後の患者、すなわちレシピエントに投与される。 Formulations of the present invention is administered patients after liver transplantation, i.e. to the recipient. また、本発明の製剤は、肝移植前の患者、すなわちドナーに肝移植前に予め投与しておいてもよい。 Further, formulations of the present invention, patients prior to liver transplantation, i.e. may have been administered in advance before liver transplantation in the donor. このような患者に投与する場合、投与患者の症状の程度、健康状態、 When administered such a patient, severity of the symptoms of patients treated, health,
年齢、体重等の条件によって異なり、特に限定されないが、成人患者1日当たり精製TCF−IIとして 0.6mg〜 Age, depends conditions, weight, etc., but are not limited to, 0.6Mg~ as adults daily purified TCF-II
600mg、好ましくは 6mg〜 60mg を含有する製剤を1日1回若しくはそれ以上投与すれば良い。 600 mg, preferably may be administered once a day or more formulations containing the 6mg~ 60mg.

【0009】さらに、本発明の製剤は、免疫抑制剤と併用することにより、その作用を増強することができる。 Furthermore, formulations of the present invention, in combination with immunosuppressive agents, it is possible to enhance the effect.
この時用いられる免疫抑制剤としては、シクロスポリン、ミゾリビン、タクロリムス水和物などが挙げられ、 As the case immunosuppressive agent used, cyclosporin, mizoribine and tacrolimus hydrate and the like,
特に好ましくはタクロリムス水和物が用いられる。 Particularly preferably tacrolimus hydrate is used. これらの投与量は、肝細胞増殖因子と同様に投与患者の条件によって異なり、特に限定されないが、成人患者1日当たり 0.1〜100mg 程度を投与するとよい。 These doses are different depending on conditions of patients administered like the hepatocyte growth factor is not particularly limited, may be administered to an adult patient about 1 day 0.1-100 mg. 投与は、肝細胞増殖因子と免疫抑制剤とを同時に一剤の形あるいは二剤の形で投与してもよいし、また、それぞれの剤を個別に投与時期を変えて投与してもよい。 Administration may be administered in the form of the form or two agents hepatocyte growth factor and an immunosuppressive agent and at the same time one agent, may also be each agent administered in varying individually administration time. 本発明ではそれらの投与形態を併用という。 That together these dosage forms in the present invention.

【0010】以下の実施例によって本発明をより詳細に説明するが、これらは単に例示したのみであり、これらによって本発明は何ら限定されるものではない。 [0010] describing the following examples the present invention in more detail, but these are merely only been illustrated, the present invention these are not intended to be limited.

【0011】 [0011]

【実施例1】 TCF−IIの精製 WO90/10651号公報に開示された方法及び東尾らの方法(Higashio,K. et al, BBRC, vol.170, pp397-404 EXAMPLE 1 TCF-II method disclosed in purified WO90 / 10651 and JP and Higashio et al. Method (Higashio, K. Et al, BBRC, vol.170, pp397-404
(1990)) に準じて細胞を培養し、精製TCF−IIを得た。 According to (1990)) cells were cultured to obtain purified TCF-II. すなわち、ヒト繊維芽細胞IMR-90 (ATCC CCL 18 That is, human fibroblast IMR-90 (ATCC CCL 18
6)細胞を5%仔牛血清を含むDMEM培地 100mlを入れたローラーボトルに3×10 6個移植し、0.5〜2回転/ 6) Cells were 3 × 10 6 cells implanted in roller bottles in DMEM medium 100ml with 5% calf serum, 0.5 to 2 revolutions /
分の速度で回転させながら7日間培養を続けた。 It was continued for 7 days of culture while rotating at a rate of. 総細胞数が1×10 7個になったところでトリプシンにより細胞を剥離し細胞をボトル底面に集め、5〜9メッシュのセラミック100g(東芝セラミック社)を殺菌して投入し、 The total number of cells collected in a detached bottle bottom cells the cells by trypsin was made to 1 × 10 7 cells were placed in sterilized 5-9 mesh ceramic 100g (Toshiba Ceramic Co., Ltd.),
24時間静置して培養した。 They were cultured for 24 hours. その後、上記培養液を 500ml Then, the above-mentioned culture medium 500ml
加え、培養を継続した。 In addition, the culture was continued. 7〜10日ごとに培地を全量回収し、新鮮培地を補給した。 The total amount recovered and the medium every 7 to 10 days, supplemented with fresh medium. このようにして2ヵ月間生産を継続し、ローラーボトル一本当たり4リットルの培養液を回収した。 In this way, to continue the two-month production, it was recovered 4 liters of culture fluid per one roller bottle. このようにして得た培養液当たりの生産量は32u/mlであった。 Production per culture thus obtained was 32u / ml. 培養液 750リットルをメンブランフィルター(MW6000カット;アミコン社)処理によりUF濃縮し、CMセファデックスC-50(ファルマシア社)、 ConAセファロース(ファルマシア社)、MonoS Culture 750 l of membrane filter; and UF concentrated by (MW6000 cut Amicon) process, CM Sephadex C-50 (Pharmacia), ConA-Sepharose (Pharmacia), MonoS
カラム(ファルマシア社)、ヘパリンセファロース(ファルマシア社)による4段階のクロマト精製を行い、精製TCF−IIを得た。 Column (Pharmacia), the chromatographic purification of 4 steps by heparin Sepharose (Pharmacia) to obtain purified TCF-II.

【0012】 [0012]

【実施例2】 遺伝子組換えTCF−IIの生産 WO92/01053号公報に開示された方法に従い、TCF−II According EXAMPLE 2 The method disclosed in Production WO92 / 01053 discloses a recombinant TCF-II, TCF-II
遺伝子を組み込んだ細胞を培養し、精製TCF−IIを得た。 Culturing the cells that have incorporated the gene to obtain purified TCF-II. すなわち、形質転換ナマルワ(Namalwa)細胞を培養し、培養液20リットルを得た。 That is, culturing the transformed Namalwa (Namalwa) cells to obtain a culture liquid 20 liter. この培養液をCM−セファデックスC−50クロマト(ファルマシア社)、Con-A The culture CM- Sephadex C-50 chromatography (Pharmacia), Con-A
セファロースCL−6Bクロマト(ファルマシア社)、 Sepharose CL-6B chromatography (Pharmacia),
MonoSカラム(ファルマシア社)を装着したHPLCの順に処理を行い、約11mgの活性TCF−IIを得た。 It performs processing in the order of HPLC equipped with MonoS column (Pharmacia) to obtain active TCF-II of about 11 mg.

【0013】 [0013]

【実施例3】 TCF−II製剤の製造前記実施例2により得られた遺伝子組換えTCF−II Example 3 TCF-II preparations gene obtained by the production in Example 2 of Recombinant TCF-II
の、注射剤の製造例を示す。 Of, showing an example of producing injections. TCF−II 20 μg ヒト血清アルブミン 100 mg 上記組成をpH 7.0の 0.01Mリン酸緩衝液(PBS)に溶解し全量を20mlに調製し、滅菌後バイアル瓶に2mlづつ分注したものを凍結乾燥後密封した。 TCF-II 20 a μg human serum albumin 100 mg The above composition was dissolved in 0.01M phosphate buffer pH 7.0 (PBS) to prepare a total volume of 20 ml, after freeze drying which was dispensed 2ml increments min in sterile after vial sealed.

【0014】 TCF−II 40 μg ツイーン80 1 mg ヒト血清アルブミン 100 mg 上記組成を注射用生理食塩水に溶解し全量を20mlに調製し、滅菌後バイアル瓶に2mlづつ分注したものを凍結乾燥後密封した。 [0014] The total amount was dissolved in TCF-II 40 μg Tween 80 1 mg Human serum albumin 100 mg The above saline for injection composition prepared 20 ml, after freeze drying which was dispensed 2ml increments min in sterile after vial sealed.

【0015】 TCF−II 20 μg ツイーン80 2 mg ソルビトール 4 g 上記組成をpH 7.0の0.01M PBSに溶解し全量を20mlに調製し、滅菌後バイアル瓶に2mlづつ分注したものを凍結乾燥後密封した。 [0015] TCF-II 20 μg Tween 80 2 mg Sorbitol 4 g The above composition was prepared on the total amount dissolved in 0.01 M PBS of pH 7.0 to 20 ml, after freeze-drying sealed those dispensed 2ml increments min in sterile after vial did.

【0016】 TCF−II 40 μg ツイーン80 1 mg グリシン 2 g 上記組成を注射用生理食塩水に溶解し全量を20mlに調製し、滅菌後バイアル瓶に2mlづつ分注したものを凍結乾燥後密封した。 [0016] The total amount was dissolved TCF-II 40 μg Tween 80 1 mg Glycine 2 g The above composition physiological saline for injection was prepared in 20 ml, was what was dispensed 2ml increments min in sterile after vials were sealed after lyophilization .

【0017】 TCF−II 40 μg ツイーン80 1 mg ソルビトール 2 g グリシン 1g 上記組成を注射用生理食塩水に溶解し全量を20mlに調製し、滅菌後バイアル瓶に2mlづつ分注したものを凍結乾燥後密封した。 [0017] The total amount was dissolved TCF-II 40 μg Tween 80 1 mg Sorbitol 2 g Glycine 1g The above composition physiological saline for injection was prepared in 20 ml, after freeze drying which was dispensed 2ml increments min in sterile after vial sealed.

【0018】 TCF−II 20 μg ソルビトール 4 g ヒト血清アルブミン 50 mg 上記組成をpH7.0 の0.01M PBSに溶解し全量を20mlに調製し、滅菌後バイアル瓶に2mlづつ分注したものを凍結乾燥後密封した。 [0018] TCF-II 20 μg Sorbitol 4 g Human serum albumin 50 mg The above composition was prepared to total dissolved 0.01 M PBS of pH7.0 to 20 ml, lyophilized those dispensed 2ml increments min in sterile after vial and post-sealed.

【0019】 TCF−II 40 μg グリシン 2 g ヒト血清アルブミン 50 mg 上記組成を注射用生理食塩水に溶解し全量を20mlに調製し、滅菌後バイアル瓶に2mlづつ分注したものを凍結乾燥後密封した。 [0019] TCF-II 40 μg glycine 2 g human serum albumin 50 mg The above composition was prepared on the total amount dissolved in physiological saline for injection to 20 ml, after freeze-drying sealed those dispensed 2ml increments min in sterile after vial did.

【0020】 TCF−II 40 μg ヒト血清アルブミン 50 mg 上記組成をpH7.0 の0.01M PBSに溶解し全量を20mlに調製し、滅菌後バイアル瓶に2mlづつ分注したものを凍結乾燥後密封した。 [0020] The total amount was dissolved in TCF-II 40 μg 0.01M PBS of human serum albumin 50 mg The above composition pH7.0 was prepared in 20 ml, was what was dispensed 2ml increments min in sterile after vials were sealed after lyophilization .

【0021】 [0021]

【実施例4】 部分肝移植における効果 Lewis 系ラット(雄、体重 235〜312 g)の肝臓の左葉及び尾状葉を切除後、UW液(ビアスパン(登録商標) EXAMPLE 4 Effect Lewis rats in the partial liver transplantation after resection (male, weighing 235-312 g) the left lobe and the caudate lobe of the liver, UW solution (Biasupan (R)
液、藤沢薬品社)に1時間冷蔵保存し、これをドナー肝とした。 Liquid, stored for 1 hour cold to Fujisawa Co.), which was used as a donor liver. このドナー肝を、肝臓を全摘出した30匹のレシピエントラット(Lewis 系ラット、雄)に同所性に移植した。 The donor liver was implanted orthotopically in Thirty recipient rats totally excised liver (Lewis rats, male). 尚、この時のドナーとレシピエントの体重差は 1 Incidentally, the weight difference between the donor and recipient at this time 1
0g以内とした。 It was within the 0g. このレシピエントラットを各15匹づつ、 Each 15 animals at a time the recipient rats,
TCF−II投与及び非投与の2群に分けた。 They were divided into two groups of TCF-II administration and non-administration. TCF−II TCF-II
投与群は移植直後から12時間毎に実施例2で得られたT Administered group obtained in Example 2 immediately after transplantation every 12 hours T
CF−II 125μg を生理食塩水中に懸濁し、これを静注した。 The CF-II 125 [mu] g was suspended in saline and injected intravenously with this. 術後1、3、7日目に犠牲死させ、肝重量の測定及び血清の採取を行った。 After surgery for 1, 3, 7 day sacrificed, was measured and serum collection of liver weight. 採取した血清は、常法により総蛋白濃度及びアルブミン濃度の測定を行った。 The collected serum was measured of the total protein concentration and the albumin concentration in a conventional manner. 結果を図1〜3に示す。 The results are shown in Figure 1-3.

【0022】この結果、TCF−II投与群は非投与群と比較して、体重10g 当たりの肝重量比、総蛋白濃度及びアルブミン濃度において、有意に上昇していた。 [0022] Consequently, TCF-II administered group as compared to the non-administration group, the liver weight per body weight 10 g, in total protein concentration and albumin concentration was significantly increased. このことから、TCF−IIの移植肝の機能改善、再生促進効果が確認された。 From this fact, functional improvement of the transplanted liver of TCF-II, regeneration promoting effect was confirmed.

【0023】 [0023]

【実施例5】 異系間部分肝移植における効果 Lewis 系ラット(雄、体重 262〜328g)の肝臓の左葉及び尾状葉を切除後、UW液(ビアスパン(登録商標) EXAMPLE 5 Effect Lewis rats in allogeneic partial liver transplantation after resection (male, body weight 262~328G) the left lobe and the caudate lobe of the liver, UW solution (Biasupan (R)
液、藤沢薬品社)に1時間冷蔵保存し、これをドナー肝とした。 Liquid, stored for 1 hour cold to Fujisawa Co.), which was used as a donor liver. このドナー肝を、肝臓を全摘出した18匹のレシピエントラット(BN系、雄、体重 262〜338g)に同所性に移植した。 The donor liver, liver total excised Eighteen recipient rats (BN type, male, body weight 262~338G) were implanted orthotopically into. 尚、この時のドナーとレシピエントの体重差は10g以内とした。 Incidentally, the weight difference between the donor and recipient at this time was within 10g. このレシピエントラットを各9匹づつ、TCF−II投与及び非投与群に分けた。 Each nine increments the recipient rats were divided into TCF-II administration and non-administration group. TC TC
F−II投与群は、実施例2で得られたTCF−IIをクエン酸緩衝液中に懸濁したものを、移植直後から12時間毎に 500μg /kgを1週間静注した。 F-II administration group, those suspended TCF-II obtained in Example 2 in citrate buffer, a 500 [mu] g / kg intravenous injection 1 week every 12 hours immediately after transplantation. 術後3及び7日目に、尾静脈より採血し血清を分離した。 In post-operative 3 and 7 days, and serum was separated blood was collected from the tail vein. 分離した血清を用い、常法により総蛋白濃度、アルブミン濃度、GO Using the separated serum, total protein concentration, albumin concentration by a conventional method, GO
T、及びGPTの測定を行った。 T, and the measurement of GPT were carried out. 結果を表1〜4に示す。 The results are shown in Tables 1-4 to.

【0024】 [0024]

【表1】 [Table 1]

【0025】 [0025]

【表2】 [Table 2]

【0026】 [0026]

【表3】 [Table 3]

【0027】 [0027]

【表4】 [Table 4]

【0028】この結果、TCF−II投与群は、非投与群と比較して総蛋白濃度及びアルブミン濃度が高く、又、 [0028] Consequently, TCF-II administration group, high total protein concentration and albumin concentration as compared to the non-administration group, also,
GOT及びGPTの上昇が抑制された。 Increase in GOT and GPT was suppressed. このことから、 From this,
TCF−IIによる蛋白合成能促進及び細胞拒絶反応に伴う肝炎抑制効果、即ち移植肝の機能改善、再生促進効果が確認された。 TCF-II by protein synthesis-enhancing and hepatitis suppression effect associated with cell rejection, i.e. improvements in transplantation liver regeneration promoting effect was confirmed.

【0029】 [0029]

【実施例6】 TCF−II処理ドナーの肝移植における効果 (1) ドナー肝の準備 Lewis 系ラット(雄、体重約300g) に、移植1週間前より実施例2で得られたTCF−IIを 500μg/kgを1日2 Example 6 TCF-II treatment effects in liver transplant donor (1) Preparation of the donor liver Lewis rats (male, body weight about 300 g) in the TCF-II obtained in Example 2 than 1 week before transplantation 500μg / kg per day 2
回投与した。 Times were administered. 1週間後に肝臓を30%摘出し、UW液(ビアスパン(登録商標)液、藤沢薬品社)に1時間冷蔵保存し、これをドナー肝とした。 The liver was excised 30% after one week, UW solution (Biasupan (registered trademark) solution, Fujisawa Co.) was stored for 1 hour refrigerated, which was used as a donor liver. (2) 部分肝移植実施例6−(1) で得られたドナー肝を、肝臓を全摘出した2匹のレシピエントラット(Lewis 系、雄、体重約 3 (2) partial donor liver obtained in liver transplantation Example 6- (1), two mice recipient rats totally excised liver (Lewis system, male, body weight about 3
00g)に同所性に移植した。 Were transplanted orthotopically to 00g). 尚、この時のドナーとレシピエントの体重差は10g 以内とした。 Incidentally, the weight difference between the donor and recipient at this time was within 10g. 又、TCF−II処理していないドナー肝を、TCF−II処理したものと同様に30%部分移植した同系のラット(3匹)を対照群とした。 Also, the donor liver that is not TCF-II treatment was TCF-II treated ones as well as 30% partial transplanted syngeneic rats (3 animals) and control group. 各群の移植後からの生存日数を指標とした。 The survival number of days from the after transplantation of each group was used as an index. 結果を表5に示す。 The results are shown in Table 5.

【0030】 [0030]

【表5】 [Table 5]

【0031】この結果、TCF−II処理肝レシピエント群は、未処理肝レシピエント群と比較して有意に生存日数が延長された。 [0031] Consequently, TCF-II treated liver recipient group, significantly survival days compared to untreated liver recipients group is extended.

【0032】 [0032]

【実施例7】 異系間部分肝移植時におけるTCF−IIと免疫抑制剤と Example 7 and TCF-II and immunosuppressant during allogeneic portion liver transplantation
の併用効果 Lewis 系ラット(雄、体重 250〜286g) の肝臓の左葉及び尾状葉を切除後、UW液(ビアスパン(登録商標) The combined effect Lewis rats after resection (male, body weight 250~286G) the left lobe and the caudate lobe of the liver, UW solution (Biasupan (R)
液、藤沢薬品社)に1時間冷蔵保存し、これをドナー肝とした。 Liquid, stored for 1 hour cold to Fujisawa Co.), which was used as a donor liver. このドナー肝を、肝臓を全摘出した30匹のレシピエントラット(BN系、雄、体重 244〜282g) に同所性に移植した。 The donor liver, Thirty recipient rats totally excised liver (BN type, male, body weight 244~282G) were implanted orthotopically into. 尚、この時のドナーとレシピエントの体重差は10g 以内とした。 Incidentally, the weight difference between the donor and recipient at this time was within 10g. このレシピエントラットを各10 Each the recipient rat 10
匹づつ、TCF−II+タクロリムス水和物(プログラフ(登録商標)、藤沢薬品社)投与群(I群)、タクロリムス水和物単独投与群(II群)、及び非投与群(III群) Animal increments, TCF-II + tacrolimus hydrate (Prograf (R), Fujisawa Co.) administered group (I group), tacrolimus hydrate alone administration group (II group), and non-administered group (III group)
に分けた。 It was divided into. I群及びII群に、タクロリムス水和物を移植直後から24時間毎に 500μg/kgを1週間筋注し、I群は、さらに実施例2で得られたTCF−IIをクエン酸緩衝液中に懸濁したものを、移植直後から12時間毎に 500 In group I and group II, tacrolimus hydrate immediately after transplantation dispensed week muscle 500 [mu] g / kg every 24 hours, Group I is further the TCF-II obtained in Example 2 citrate buffer 500 those suspended, immediately after transplantation every 12 hours
μg/kgを1週間静注した。 The μg / kg was injected intravenously for one week. 術後7日目に、犠牲死させ、 In post-operative day 7, it was sacrificed,
血清の採取を行った。 The serum collection was carried out. 採取した血清は、常法により総蛋白濃度、血中アルブミン濃度、GOT、及びGPTの測定を行った。 The collected serum was performed total protein concentration by a conventional method, blood albumin concentration, GOT, and the measurement of GPT. 結果を表6に示す。 The results are shown in Table 6.

【0033】 [0033]

【表6】 [Table 6]

【0034】この結果、I群及びII群はいずれにおいてもIII 群と比較して、総蛋白濃度及びアルブミン濃度が上昇し、GOT及びGPTの上昇が抑制された。 [0034] As a result, compared with even Group III in any group I and group II, the total protein concentration and albumin concentration increases, increase in GOT and GPT was suppressed. また、 Also,
その程度はI群でより大きかった。 The extent was greater in group I. このことから、肝移植後の免疫抑制剤投与時にTCF−IIを併用投与することにより、蛋白合成能促進及び細胞性拒絶反応に伴う肝炎抑制効果、即ち移植肝の機能改善、再生促進効果が増強することが確認された。 Therefore, by co-administration of TCF-II during immunosuppressant administration after liver transplantation, hepatitis suppression effects associated with protein synthesis-enhancing and cellular rejection, i.e. improvements in transplantation liver regeneration promoting effect enhanced it was confirmed that.

【0035】 [0035]

【発明の効果】以上の結果より、本発明により、肝細胞増殖因子を有効成分とする移植肝機能改善・再生促進剤、及び肝細胞増殖因子と免疫抑制剤とを併用した移植肝機能改善・再生促進剤が提供される。 From the above results, according to the present invention, the present invention, implanted improving liver function and regeneration promoter having for an active ingredient a hepatocyte growth factor, and transplanted liver function improving and using a combination of hepatocyte growth factor and an immunosuppressive agent Play accelerator is provided. 本発明における肝細胞増殖因子としては、特にTCF−IIが好ましい。 The hepatocyte growth factor in the present invention, especially TCF-II are preferred.
本発明により、移植した肝組織の機能改善及び再生が促進され、肝移植手術後の肝機能を早期に回復することができる。 The present invention is promoted functional improvement and regeneration of the transplanted liver tissue can recover quickly liver function after surgery liver transplantation.

【図面の簡単な説明】 BRIEF DESCRIPTION OF THE DRAWINGS

【図1】実施例4のTCF−II投与群(TCF) 及び非投与群(CONT)における、10g 当たりの肝重量の比較を示す。 [1] TCF-II administered group Example 4 (TCF) and non-administration group in (CONT), shows a comparison of liver weight per 10g.

【符号の説明】 DESCRIPTION OF SYMBOLS

* P<0.05で有意差あり ** P<0.01で有意差あり * There is a significant difference at P <0.05 there is a significant difference in the ** P <0.01

【図2】実施例4のTCF−II投与群(TCF) 及び非投与群(CONT)における、血清総蛋白濃度の比較を示す。 [Figure 2] TCF-II administered group Example 4 (TCF) and non-administration group in (CONT), shows a comparison of serum total protein concentration.

【符号の説明】 DESCRIPTION OF SYMBOLS

* P<0.05で有意差あり ** P<0.01で有意差あり * There is a significant difference at P <0.05 there is a significant difference in the ** P <0.01

【図3】実施例4のTCF−II投与群(TCF) 及び非投与群(CONT)における、血清アルブミン濃度の比較を示す。 [Figure 3] TCF-II administered group Example 4 (TCF) and non-administration group in (CONT), shows a comparison of serum albumin concentration.

【符号の説明】 DESCRIPTION OF SYMBOLS

* P<0.05で有意差あり ** P<0.01で有意差あり * There is a significant difference at P <0.05 there is a significant difference in the ** P <0.01

Claims (3)

    【特許請求の範囲】 [The claims]
  1. 【請求項1】 肝細胞増殖因子を有効成分とする移植肝機能改善・再生促進剤。 1. A graft improving liver function and regeneration promoter having for an active ingredient a hepatocyte growth factor.
  2. 【請求項2】 肝細胞増殖因子と免疫抑制剤とを併用する移植肝機能改善・再生促進剤。 Wherein hepatocyte growth factor and an immunosuppressive agent and transplanted liver function improving and regeneration-promoting agent used in combination.
  3. 【請求項3】 肝細胞増殖因子がTCF−IIである、請求項1または2記載の移植肝機能改善・再生促進剤。 Wherein hepatocyte growth factor is TCF-II, according to claim 1 or 2 transplantation improving liver function and regeneration promoters described.
JP9168177A 1996-06-10 1997-06-10 Accelerator for amelioration and regeneration of transplanted hepatic function Pending JPH10194986A (en)

Priority Applications (5)

Application Number Priority Date Filing Date Title
JP17055596 1996-06-10
JP8-170555 1996-11-12
JP31553296 1996-11-12
JP8-315532 1996-11-12
JP9168177A JPH10194986A (en) 1996-06-10 1997-06-10 Accelerator for amelioration and regeneration of transplanted hepatic function

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP9168177A JPH10194986A (en) 1996-06-10 1997-06-10 Accelerator for amelioration and regeneration of transplanted hepatic function

Publications (1)

Publication Number Publication Date
JPH10194986A true JPH10194986A (en) 1998-07-28

Family

ID=27322970

Family Applications (1)

Application Number Title Priority Date Filing Date
JP9168177A Pending JPH10194986A (en) 1996-06-10 1997-06-10 Accelerator for amelioration and regeneration of transplanted hepatic function

Country Status (1)

Country Link
JP (1) JPH10194986A (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005298392A (en) * 2004-04-09 2005-10-27 Shinji Iwasaki Medicinal composition for prophylaxis or treatment of rejection after liver transplantation
WO2006077675A1 (en) * 2005-01-24 2006-07-27 Kringle Pharma Inc. Fibrosis inhibitor for implanted organ
US7115568B2 (en) 1997-03-14 2006-10-03 Daiichi Pharmaceutical Co., Ltd. Methods using TCF II
US7306791B2 (en) 1997-03-11 2007-12-11 Daiichi Sankyo Co., Ltd. Agent for preventing and/or treating multiple organ failure

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7306791B2 (en) 1997-03-11 2007-12-11 Daiichi Sankyo Co., Ltd. Agent for preventing and/or treating multiple organ failure
US7115568B2 (en) 1997-03-14 2006-10-03 Daiichi Pharmaceutical Co., Ltd. Methods using TCF II
US7138372B2 (en) 1997-03-14 2006-11-21 Daiichi Pharmaceutical Co., Ltd. Agent for preventing and/or treating cachexia
JP2005298392A (en) * 2004-04-09 2005-10-27 Shinji Iwasaki Medicinal composition for prophylaxis or treatment of rejection after liver transplantation
WO2006077675A1 (en) * 2005-01-24 2006-07-27 Kringle Pharma Inc. Fibrosis inhibitor for implanted organ
US7696170B2 (en) 2005-01-24 2010-04-13 Kringle Pharma Inc. Fibrosis inhibitor for implanted organ
US8076289B2 (en) 2005-01-24 2011-12-13 Kringle Pharma Inc. Fibrosis inhibitor for implanted organ
US8383588B2 (en) 2005-01-24 2013-02-26 Kringle Pharma Inc. Fibrosis inhibitor for implanted organ

Similar Documents

Publication Publication Date Title
Macdougall et al. Clinical pharmacokinetics of epoetin (recombinant human erythropoietin)
CA1336680C (en) Method and compositions for the treatment and prevention of septic shock
EP0820299B1 (en) Formulations for il-12
EP1983975B1 (en) Method of treatment for muscular dystrophy
KR100231398B1 (en) Stabilized factor viii preparation
US4760051A (en) Use of GHL-Cu as a wound-healing and anti-inflammatory agent
ES2258154T3 (en) Method for obtaining a protein fraction enriched in TGF-beta in activated form, protein fraction and therapeutic applications.
EP0639079B1 (en) Methods for treating interleukin-1 and tumor necrosis factor mediated diseases
Gorantla et al. Immunosuppressive agents in transplantation: Mechanisms of action and current anti‐rejection strategies
Donohue et al. The fate of interleukin-2 after in vivo administration.
Granholm et al. NGF and anti-transferrin receptor antibody conjugate: short and long-term effects on survival of cholinergic neurons in intraocular septal transplants.
CN1089344C (en) Inhibitor of stem cell proliferation and uses thereof
US20050277106A1 (en) Methods and active substances for protecting organs
Calne et al. Combined immunosuppressive action of phytohaemagglutinin and azathioprine (Imuran) on dogs with renal homotransplants
JP3763479B2 (en) Biologically active TGF-.beta.1 and TGF-.beta.2 peptide
CN1097058C (en) Platelet growth accelerator
DE60219611T2 (en) Modified annexin-proteins and prevention and treatment of thrombosis
EP0213180B1 (en) Neovascularization inhibitors and methods for their production and use
JP3866197B2 (en) A therapeutic agent for ischemic disease
EP0816381A1 (en) Peg-modified hgf
KR100415636B1 (en) A pharmaceutical composition for the treatment of multiple sclerosis comprising interferon consensus and interleukin-1 receptor antagonist
EP1004312A1 (en) Method for treating multiple sclerosis
JP2966592B2 (en) Human monoclonal antibody formulations stabilized
WO1994015625A1 (en) Factor viii - polymeric conjugates
WO1993011793A1 (en) Use of the combination of anti-tumor necrosis factor plus interleukin-6 to treat septic shock

Legal Events

Date Code Title Description
A521 Written amendment

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20040326

A621 Written request for application examination

Free format text: JAPANESE INTERMEDIATE CODE: A621

Effective date: 20040326

A131 Notification of reasons for refusal

Free format text: JAPANESE INTERMEDIATE CODE: A131

Effective date: 20070823

A02 Decision of refusal

Free format text: JAPANESE INTERMEDIATE CODE: A02

Effective date: 20071211