JP4463885B2 - Fulminant hepatitis disease treatment - Google Patents

Description

【００２４】
試験例２
ＨＧＦ投与によるアポトーシス肝障害の抑制効果
ＨＧＦがFas誘起アポトーシスによる肝障害を抑制できるかどうか、in vivoで検討した。
材料としてJo-2抗体（Fas抗体）、BALB/Cマウス雄（８週令）及び組換えヒトＨＧＦ(human recombinant ＨＧＦ)を使用した。方法は以下のとおりである。
Jo-2抗体投与量として、マウス当たり5μgを腹腔内（i.p.）投与（マウス９匹）した。Jo-2抗体投与６時間前に、ＨＧＦ（マウス４匹）あるいはコントロールとして食塩水（マウス５匹）をi.p.投与した。さらに、抗体投与時、抗体投与６時間後、１２時間後に連続してＨＧＦあるいは食塩水のi.p.投与を行った。Jo-2抗体投与１２時間後に尾静脈より採血し、２４時間後に剖検、組織を採取した。検討項目としては、肝機能マーカーとして、GPTを測定し、肝臓の一部を１０％ホルマリン固定して、顕微鏡下Ｈ＆Ｅ染色による肝障害の判定、さらにアポトーシスの評価を行った。
【００２５】
結果を表２並びに図２及び３（病理組織標本の顕微鏡写真）に示す。なお、図２はＨＧＦ投与群のマウスの肝の病理組織標本の顕微鏡写真であり、Ａは試験Ｎｏ．１６のマウスの肝の病理組織標本の顕微鏡写真（倍率２００倍）、Ｂは試験Ｎｏ．１９のマウスの肝の病理組織標本の顕微鏡写真（倍率１００倍）である。また、図３は食塩水投与群（コントロース）のマウス（試験Ｎｏ．１４のマウス）の肝の病理組織標本の顕微鏡写真であり、Ａは倍率１００倍、Ｂは倍率２００倍である。
表２に示されたように、ＨＧＦ投与マウスは４例とも肝機能GPTが低値を示した。また、図２に示されるように、アポトーシスによる肝障害は明らかに抑制された。一方、ＨＧＦの代わりに食塩水を投与したマウスは１例がJo-2抗体投与後８時間に死亡したが、その肝組織は広範性びまん性にほとんどの肝細胞がアポトーシスによる壊死の状態を呈しており、肝不全による死亡であった。また、残りの４例でも肝機能GPTが高値を示した。更に、図３に示されるように、門脈域周囲(Zone 1)から中間帯(Zone 2)にかけて、著明な肝細胞のアポトーシスとそれによる細胞死という肝病理組織像を呈する重度の肝障害を伴っていた。
以上の結果から、ＨＧＦはFas抗体によって誘起されるアポトーシス肝障害を抑制することが認められた。
【００２６】
【表２】

【００２７】
実施例１
生理食塩水１００ｍｌ中にＨＧＦ１ｍｇ、マンニトール１ｇ及びポリソルベート８０ １０ｍｇを含む溶液を無菌的に調製し、１ｍｌずつバイアルに分注した後、凍結乾燥して密封することにより凍結乾燥製剤を得た。
【００２８】
実施例２
０．０２Ｍリン酸緩衝液（０．１５Ｍ ＮａＣｌ及び０．０１％ポリソルベート８０含有、ｐＨ７．４）１００ｍｌ中にＨＧＦ１ｍｇ及びヒト血清アルブミン１００ｍｇを含む水溶液を無菌的に調製し、１ｍｌずつバイアルに分注した後、凍結乾燥して密封することにより凍結乾燥製剤を得た。
【図面の簡単な説明】
【図１】試験例１のマウスの肝の病理組織標本の顕微鏡写真の一例を示すものである（生物の形態）。図において、Ａ及びＢは倍率１００倍、Ｃは倍率２００倍の顕微鏡写真である。
【図２】試験例２におけるＨＧＦ投与群のマウスの肝の病理組織標本の顕微鏡写真である（生物の形態）。図において、Ａは試験Ｎｏ．１６のマウスの肝の病理組織標本の顕微鏡写真（倍率２００倍）、Ｂは試験Ｎｏ．１９のマウスの肝の病理組織標本の顕微鏡写真（倍率１００倍）である。
【図３】試験例２における食塩水投与群（コントロール）のマウス（試験Ｎｏ．１４のマウス）の肝の病理組織標本の顕微鏡写真である（生物の形態）。図において、Ａは倍率１００倍、Ｂは倍率２００倍である。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a therapeutic agent for fulminant hepatitis. More specifically, the present invention relates to a therapeutic agent for fulminant hepatitis, which contains hepatocyte growth factor (hereinafter referred to as HGF) as an active ingredient and is useful for the treatment and prevention of fulminant hepatitis.
[0002]
[Prior art]
The liver is essential for living organisms with many physiological functions such as gluconeogenesis, amino acid metabolism, lipid metabolism, plasma protein synthesis, secretion, production and secretion of bile, detoxification, storage of sugar as an energy source, storage of vitamins, etc. It is an organ. This organ develops hepatitis due to viral infection and prolonged alcohol consumption.
Fulminant hepatitis is a severe liver disorder that causes hepatic necrosis and shedding of hepatocytes due to hepatitis viruses and drugs, and has a high fatality rate. According to the diagnostic criteria of the 12th Inuyama Symposium (August 1981), severe hepatitis refers to hepatic coma based on severe liver dysfunction within about 8 weeks after onset of symptoms of hepatitis, and a prothrombin time of 40 %, And there are an acute type in which encephalopathy develops within 10 days after onset and a subacute type that develops after 11 days. According to the clinical picture, jaundice is advanced within a few days after the onset of hepatitis symptoms, and it is rapidly accompanied by disturbance of consciousness. There is a tendency to bleed from an early stage, and there is a reduction in the cloudy sound field. The serum total bilirubin level is often 10 mg / dl or more, and the serum transaminase level shows a markedly high value in the initial stage, but falls significantly within several days. The prothrombin value shows a remarkable decrease of 40% or less. Cerebral edema, gastrointestinal bleeding, renal failure, intravascular coagulation syndrome, systemic infections are likely to accompany these complications and often cause death. The prognosis is extremely poor, and the survival rate in Japan is 20-30%. As for treatment, there is no established treatment, and systemic management is required at present.
[0003]
Plasma exchange, exchange transfusion, artificial liver assist device, and glucagon / insulin therapy are performed. Thus, it is no exaggeration to say that there is no therapeutic agent for treating this fulminant hepatitis. Although plasma exchange is performed at about 80% in Japan, there is almost no difference in survival rate between the enforcement group and the non-treatment group. In addition, glucagon / insulin therapy is also performed in 90 cases, but there is not much difference in survival rate between the administration group and the non-treatment group. Special composition amino acid administration is also performed in most cases, but there is almost no difference in survival rate between the two groups. In other words, 80% of the cases have undergone plasma exchange, glucagon / insulin therapy, and special composition amino acid therapy, which are typical treatments in Japan, but the survival rate is almost the same as before such treatment. It is said that it has not risen, and there is no choice but to be very skeptical about its effects.
[0004]
[Problems to be solved by the invention]
As for fulminant hepatitis, according to the diagnostic criteria of the 1981 Inuyama Symposium, hepatitis II and prothrombin time (PT) increased to 40% based on severe liver dysfunction within 8 weeks after hepatitis manifestation. It is defined as a decreasing case.
Since the liver is an organ with a large reserve capacity, if about 30% remains, it is usually compensated sufficiently. However, below that, liver failure is a failure of liver function.
Specifically, the function of the liver can be broadly divided into two functions: a function for synthesizing substances and a function for metabolizing substances. Due to the failure of the function, PT is reduced as an expression of decreased synthetic ability. PT is a test that measures the function of all five coagulation factors, I, II, V, VII, and X, all of which are made in the liver. Among them, in particular, Factor VII has a short half-life and is halved in about several hours. That is, when synthetic ability falls, factor VII falls from the beginning, and PT time containing it falls. The other is coma due to a decrease in detoxification metabolic function. The reason for setting the degree of II or higher was that the degree of degree II or higher was considered significant because the subjectivity of the attending physician entered the judgment.
[0005]
The cause of the disease is hepatitis in Japan. Hepatitis includes both viral and drug hepatitis. In foreign countries, fulminant hepatitis is considered as a disease of the category of acute hepatitis, but in Japan there are many HBV carriers, and fulminant carriers are rarely seen. In the case of carriers, the onset time of hepatitis is unknown, and as long as the virus is present, chronic hepatitis may be present, and it may become fulminant.
On the other hand, in foreign countries such as the UK, viruses are almost half and slightly more than half are pharmaceutical. In particular, in the UK, there are many fulminant hepatitis caused by the use of the analgesic antipyretic drug paracetamol, that is, acetaminofeton, for the purpose of suicide. Usually, drug-induced liver damage experienced clinically is rather allergic hepatitis. In this case, the dose is too low and liver damage occurs, and even if the causative drug is removed, the inflammation maintenance mechanism is already activated. The liver damage cannot be immediately terminated.
[0006]
It is said that there are two conditions for the mechanism by which hepatitis becomes fulminant. One is that many hepatocytes are infected. The other is that there is a strong cytotoxic T lymphocyte immune response that actively destroys hepatocytes. In other words, fulminant hepatitis does not occur unless there are two factors: that many hepatocytes are infected and that the immune response is intense and hepatocyte destruction is severe. Based on this theory, hepatitis A virus is eliminated quickly, and as it spreads to the point where there is an infection, the elimination reaction occurs and is eliminated, and it is cured as acute hepatitis without becoming fulminant. In the case of hepatitis C virus, infection is advanced to many hepatocytes, and the virus does not elicit the host's immune response so strongly that it is difficult to become fulminant. It is understood that hepatitis B is most likely to become fulminant because it is intermediate and spreads with viruses and has a strong immune response.
[0007]
In 1972, pathologist Kerr et al. Described a form of cell death characterized by "nuclear aggregation and fragmentation" and called it apoptosis. Since then, cell death has been divided into two categories: necrosis and apoptosis. Necrosis is caused by pathological and non-physiological factors such as viruses, ischemia, and toxins. Morphologically, it shows an image of cell membrane breakdown after swelling of cytoplasm, nucleus, and mitochondria. On the other hand, apoptosis is a physiological death, and it is considered that cells are caused by the activation of a self-death device incorporated in the cell by comprehensively judging information by itself. It shows the morphological characteristics of nuclear enrichment and fragmentation and the biochemical characteristics of DNA fragmentation. In other words, death controlled by a gene is apoptosis, and death not controlled by a gene is necrosis.
In recent years, analysis at the gene level of apoptosis has progressed due to advances in molecular biology, and the relationship with apoptosis diseases has also attracted attention. In the liver, the acidophilic body often seen in viral hepatitis, etc. is actually apoptosis of hepatocytes. It has been shown that it is apoptotic, and it has been suggested that apoptosis is involved in many liver diseases. In particular, the Fas / Fas ligand system is the most studied as a system that induces apoptosis.In 1993, Nagata et al. Reported that intraperitoneal administration of anti-Fas antibody to mice caused extensive hepatocyte death. The possibility that the Fas system could cause fulminant hepatitis was shown (Nature, 364 , 806, 1993).
In any case, 90% or more of fulminant hepatitis is viral in Japan, the fatality rate is 50% or more in the acute type, and the fatality rate exceeds 90% in the subacute type. In addition, it is a disease with a poor prognosis, and elucidation of its cause and development of an effective treatment method are eagerly desired.
[0008]
From these viewpoints, the present inventors have found that HGF has an action of suppressing, improving or preventing liver damage caused by apoptosis, that is, Fas-induced apoptosis and accompanying liver damage, and HGF prevents fulminant hepatitis disease.・ It proved useful for treatment.
The above HGF is a protein found as a factor for growing hepatocytes in vitro (Biochem. Biophys. Res. Commun., 122 , 1450, 1984, Proc. Natl. Acad. Sci. USA, 83 , 6489, 1986, FEBS Letter, 22 , 311, 1987, Nature, 342 , 440, 1989, Proc. Natl. Acad. Sci. USA, 87 , 3200, 1990). HGF, which was discovered as a factor that specifically proliferates liver parenchymal cells, exhibits various activities such as tissue injury healing in vivo according to recent research results by many researchers including the present inventors. As a result, it is expected to be applied not only as a research target but also to therapeutic drugs for humans and animals.
[0009]
It has been elucidated that HGF is mainly produced by mesenchymal cells, and it has been clarified that a so-called paracrine mechanism in which HGF is supplied from neighboring cells as needed is established. However, when the liver or kidney is injured, the production of HGF is also increased in non-injured organs, such as the lung, so it is considered that HGF is also supplied by the so-called endocrine mechanism.
Regarding such HGF receptors, recent studies have established that the c-Met proto-oncogene encodes the HGF receptor (Bottaro et al., Science 251 , 802-804, 1991). Naldini et al., Oncogene 6 , 501-504, 1991).
As described above, a lot of knowledge about HGF has been obtained. However, it is a new knowledge that has not been known so far that HGF can suppress, improve, or prevent liver damage caused by apoptosis. This invention is made | formed based on this knowledge, and this invention aims at providing the novel therapeutic agent of fulminant hepatitis disease.
[0010]
[Means for Solving the Problems]
The present invention made to solve the above problems
(1) A therapeutic agent for fulminant hepatitis, characterized by containing HGF as an active ingredient;
(2) The therapeutic agent for fulminant hepatitis described in (1) above, wherein the fulminant hepatitis disease is a fulminant hepatitis mainly composed of liver damage caused by apoptosis.
[0011]
As the HGF used in the present invention, those prepared by various methods can be used as long as they are purified to the extent that they can be used as pharmaceuticals.
Various methods are known as methods for preparing HGF. For example, extracted and purified from liver, spleen, lung, bone marrow, brain, kidney, placenta and other organs such as rats, cows, horses and sheep, blood cells such as platelets and leukocytes, plasma and serum (See FEBS Letters, 224 , 312, 1987, Proc. Natl. Acad. Sci. USA, 86 , 5844, 1989, etc.).
[0012]
Further, HGF can also be obtained by culturing primary cultured cells or established cells that produce HGF, and separating and purifying them from the culture (culture supernatant, cultured cells, etc.). Alternatively, the gene encoding HGF may be incorporated into an appropriate vector by genetic engineering techniques, transformed into a suitable host, and the desired recombinant HGF may be obtained from the culture supernatant of the transformant. (See, for example, Nature, 342 , 440, 1989, JP-A-5-111383, Biochem. Biophys. Res. Commun., 163 , 967, 1989, etc.). The host cell is not particularly limited, and various host cells conventionally used in genetic engineering techniques such as Escherichia coli, Bacillus subtilis, yeast, filamentous fungi, plant or animal cells can be used.
[0013]
More specifically, as a method for extracting and purifying HGF from living tissue, for example, carbon tetrachloride is intraperitoneally administered to a rat, and the liver of a rat in a hepatitis state is extracted, pulverized, pulverized, It can be purified by usual protein purification methods such as gel column chromatography such as heparin sepharose and HPLC.
In addition, using gene recombination methods, animal cells such as Chinese hamster ovary (CHO) cells, mouse C127 cells, monkeys are expressed by an expression vector in which a gene encoding the amino acid of human HGF is incorporated into a vector such as bovine papillomavirus DNA. COS cells can be transformed and obtained from the culture supernatant.
[0014]
As long as the HGF thus obtained has substantially the same effect as HGF, a part of its amino acid sequence is deleted or substituted by another amino acid, a part of the other amino acid sequence is inserted, the N-terminal And / or one or more amino acids may be bonded to the C-terminus, or the sugar chain may be similarly deleted or substituted.
[0015]
The therapeutic agent of the present invention contains the above-mentioned HGF as an active ingredient, and HGF suppresses, improves, or prevents HGF against liver damage caused by apoptosis by proliferating hepatocytes as shown in the following test examples. Have Further, HGF has the advantage that it has no risk of causing side effects because it does not act on tissues that have not been damaged, but only acts on tissues that have been damaged. Therefore, the therapeutic agent of the present invention is effective for the treatment and prevention of fulminant hepatitis disease, particularly lesions mainly composed of hepatocyte proliferation.
The therapeutic agent of the present invention is applied to the treatment and prevention of various diseases caused by liver damage in various mammals (for example, cattle, horses, pigs, sheep, dogs, cats, etc.) in addition to humans.
[0016]
The therapeutic agent of the present invention can take various preparation forms (for example, liquids, solids, capsules, etc.). Generally, only the active ingredient HGF or a conventional carrier and an injection, inhalant, suppository are used. Or oral. The injection can be prepared by a conventional method, for example, HGF is dissolved in an appropriate solvent (for example, sterilized water, buffer solution, physiological saline, etc.), then filtered through a filter or the like, and sterilized. It can then be prepared by filling a sterile container. The HGF content in the injection is usually adjusted to about 0.0002-0.2 (w / v%), preferably about 0.001-0.1 (w / v%). Oral drugs are formulated into dosage forms such as tablets, granules, powders, soft or hard capsules, solutions, emulsions, suspensions, syrups, etc. It can be prepared according to Suppositories can also be prepared by a conventional method using a conventional base (for example, cacao butter, lauric butter, glycerogelatin, macrogol, witepsol). Inhalants can also be prepared according to conventional means on pharmaceutical preparations.
The HGF content in the preparation can be appropriately adjusted according to the dosage form, applicable disease and the like.
[0017]
In formulating, a stabilizer is preferably added, and examples of the stabilizer include albumin, globulin, gelatin, glycine, mannitol, glucose, dextran, sorbitol, ethylene glycol and the like. Furthermore, the preparation of the present invention may contain additives necessary for formulation, such as excipients, solubilizers, antioxidants, soothing agents, tonicity agents and the like. In the case of a liquid preparation, it is desirable to store it after removing moisture by freeze storage or freeze drying. The freeze-dried preparation is used by re-dissolving and adding distilled water for injection at the time of use.
The therapeutic agent of the present invention can be administered by an appropriate administration route according to the formulation form. For example, it can be administered in the form of an injection into veins, arteries, subcutaneous, intramuscular or the like. The dosage is appropriately adjusted according to the patient's symptoms, age, body weight, etc., but is usually 0.05 mg to 500 mg, preferably 1 mg to 100 mg as HGF, which is administered once or several times a day. It is appropriate to do.
[0018]
【The invention's effect】
In the present invention, HGF, which is an active ingredient, has an action of suppressing, improving or preventing liver damage caused by apoptosis, which is the cause of fulminant hepatitis, that is, Fas-induced apoptosis and accompanying liver damage. Therefore, the therapeutic agent of the present invention is effective for the treatment and prevention of the aforementioned fulminant hepatitis disease. Furthermore, since HGF acts only on damaged tissues, there is an effect that a drug with few side effects can be obtained.
[0019]
【Example】
EXAMPLES Hereinafter, although this invention is demonstrated in detail based on an experiment example and an Example, this invention is not limited to these examples.
Test example 1
Basic condition setting for apoptotic liver injury <br/> In the test of hepatocyte death due to apoptosis examined by Nagata et al., Using 3-5 week old BALB / C mice, Jo-2 antibody (Fas antibody) 100μg And 10 μg are administered intraperitoneally (Nature, 364 , 806, 1993). In this test, the optimal antibody dose for determining the inhibitory effect of apoptotic liver injury by HGF administration was set, and the basic conditions were set to minimize the variation among individuals with apoptotic liver injury.
As materials, Jo-2 antibody (Fas antibody) and BALB / C mouse male (8 weeks old) were used. The method is as follows.
[0020]
As a dose of Jo-2 antibody, 5 μg of Jo-2 antibody per mouse was administered intraperitoneally (ip) (2 mice), and 5 μg of Jo-2 antibody (2 mice), 2 μg (3 mice), 1 μg (mouse) 3 mice) were administered via the tail vein (iv), and as a control, saline was administered iv (1 mouse). Jo-2 antibody or saline was administered, blood was collected from the tail vein 6 hours later, and necropsy and tissue were collected 24 hours later. As examination items, GPT, TP, ALB, GOT were measured as liver function markers, a part of the liver was fixed with 10% formalin, liver damage was determined by H & E staining under a microscope, and apoptosis was evaluated. . A part of the liver was frozen, extracted with DNA, and subjected to DNA fragmentation analysis by electrophoresis. Analysis of DNA extraction and DNA fragmentation is as follows.
[0021]
1) and homogenized with a cell lysis buffer of the frozen liver 20-30mg 2ml (320mM Saccharose, 5mM MgCl 2, 10mM Tris-HCl, 1% Triton X-100, pH 7.5).
2) 100 μl of 20 mg / ml Protease was added and incubated at 50 ° C. for 2 hours.
3) RNase A was added to 200 μg / ml and further incubated at 50 ° C. for 1 hour.
4) 500 μl was taken and precipitated with isopropanol, washed with ethanol, and dissolved in 20 μl TE buffer.
5) The entire amount (20 μl) was electrophoresed with ethidium bromide-containing 2% agarose, and DNA fragmentation was observed.
[0022]
The results are shown in Table 1 and FIG. 1 (micrograph of pathological tissue specimen).
As shown in Table 1, 5 μg iv or ip administration was optimal as a result of setting the basic conditions.
FIG. 1 shows an example of a micrograph of a pathological specimen of liver. In the figure, A and B are photomicrographs at a magnification of 100 times and C at a magnification of 200 times. As shown in the figure, in the pathological tissue of the liver, apoptosis induced by administration of Jo-2 antibody occurs from Zone 1 (hepatocytes around the portal vein region), and from Zone 1 due to an increase in the amount of Jo-2 antibody. Apoptosis extends to Zone 2, Zone 3 (hepatocytes around the central vein), the degree of necrosis progresses, and it has been found that almost all hepatocytes fall into necrosis due to apoptosis widely and diffusely.
[0023]
[Table 1]

[0024]
Test example 2
Inhibitory effect of HGF administration on apoptotic liver injury It was examined in vivo whether HGF can suppress liver injury caused by Fas-induced apoptosis.
As materials, Jo-2 antibody (Fas antibody), BALB / C mouse male (8 weeks old) and recombinant human HGF (human recombinant HGF) were used. The method is as follows.
As a dose of Jo-2 antibody, 5 μg per mouse was intraperitoneally (ip) administered (9 mice). Six hours before administration of Jo-2 antibody, HGF (4 mice) or saline (5 mice) was administered ip as a control. Furthermore, at the time of antibody administration, ip administration of HGF or saline was performed continuously 6 hours and 12 hours after antibody administration. Blood was collected from the tail vein 12 hours after administration of Jo-2 antibody, and necropsy and tissue were collected 24 hours later. As examination items, GPT was measured as a liver function marker, a part of the liver was fixed with 10% formalin, liver damage was determined by H & E staining under a microscope, and apoptosis was further evaluated.
[0025]
The results are shown in Table 2 and FIGS. 2 and 3 (micrographs of pathological tissue specimens). 2 is a micrograph of a histopathological specimen of the liver of a mouse in the HGF administration group. 16 is a micrograph (magnification 200 times) of a histopathological specimen of the liver of 16 mice. It is a microscope picture (magnification 100 times) of the pathological tissue sample of the liver of 19 mice | mouths. 3 is a photomicrograph of a liver histopathological specimen of a saline-administered group (controse) mouse (mouse of test No. 14), A is 100 times magnification and B is 200 times magnification.
As shown in Table 2, all four HGF-treated mice showed low liver function GPT. In addition, as shown in FIG. 2, liver damage due to apoptosis was clearly suppressed. On the other hand, one mouse that received saline instead of HGF died 8 hours after the administration of the Jo-2 antibody, but the liver tissue was diffuse and most hepatocytes were necrotic due to apoptosis. He died of liver failure. In the remaining 4 cases, liver function GPT was also high. Furthermore, as shown in FIG. 3, severe hepatic disorder presenting a hepatic histopathological image of significant hepatocyte apoptosis and resulting cell death from the portal zone (Zone 1) to the intermediate zone (Zone 2). Was accompanied.
From the above results, it was confirmed that HGF suppresses apoptotic liver injury induced by Fas antibody.
[0026]
[Table 2]

[0027]
Example 1
A solution containing 1 mg of HGF, 1 g of mannitol and 10 mg of polysorbate 80 in 100 ml of physiological saline was prepared aseptically, dispensed into vials in 1 ml portions, freeze-dried and sealed to obtain a freeze-dried preparation.
[0028]
Example 2
An aqueous solution containing 1 mg of HGF and 100 mg of human serum albumin in 100 ml of 0.02 M phosphate buffer (containing 0.15 M NaCl and 0.01% polysorbate 80, pH 7.4) is aseptically prepared, and 1 ml is dispensed into vials. After that, freeze-dried and sealed to obtain a freeze-dried preparation.
[Brief description of the drawings]
FIG. 1 shows an example of a micrograph of a histopathological specimen of the liver of a mouse of Test Example 1 (organism form). In the figure, A and B are photomicrographs at a magnification of 100 times and C at a magnification of 200 times.
FIG. 2 is a photomicrograph of a histopathological specimen of a liver of a mouse in an HGF administration group in Test Example 2 (organism form). In FIG. 16 is a micrograph (magnification 200 times) of a histopathological specimen of the liver of 16 mice. It is a microscope picture (magnification 100 times) of the pathological tissue sample of the liver of 19 mice | mouths.
3 is a photomicrograph of a histopathological specimen of the liver of a saline-administered group (control) mouse (Test No. 14 mouse) in Test Example 2 (form of organism). In the figure, A is 100 times magnification and B is 200 times magnification.

Claims (1)

A therapeutic agent for fulminant hepatitis, comprising human HGF as an active ingredient.

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