JP3127484B2 - Hepatitis treatment - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a therapeutic agent for hepatitis.

[0002]

BACKGROUND OF THE INVENTION Hepatitis is caused by large or long-term ingestion of hepatitis A, B, and C viruses and drugs.
In such a case, it is known that liver function is significantly reduced due to reduction of liver mitochondrial function and hepatocyte necrosis. Particularly in acute hepatitis and fulminant hepatitis, plasma transaminase is significantly increased due to extensive hepatocyte necrosis. It is also known that when these symptoms become chronic, the necrotic lesions fibrillate, followed by liver fibrosis or cirrhosis.

[0003] Interferon therapy as an antiviral therapy has been used for the treatment of such liver disorders. However, interferon has not been found to have a direct effect on improving mitochondrial function or liver regeneration. Glutathione, tathione for injection (Yamanouchi Pharmaceutical Co., Ltd.) and potent neominophagency (Minophagen), which are commonly used for hepatitis, have the effect of improving plasma transaminase levels, but have poor liver mitochondrial function and liver regeneration effects. . In addition, these drugs are injections, and development of drugs that are effective even by oral administration is expected.

On the other hand, in acute liver injuries such as viral hepatitis and drug-induced hepatitis, and diseases in which these have become chronic, the plasma transaminase concentration is often used as an index for determining the condition of a patient. Therefore, the conventional therapeutic agents for hepatitis, for example, potent neominophagen C (Minophagen) has a main effect of suppressing plasma transaminase elevation by protecting cell membranes, and has a clear effect on improvement of liver function and liver regeneration. Absent. The interferon is virus removal that cause in the case of viral hepatitis, glutathione has been recognized the free-radical removal as efficacy. However, these drugs only remove the cause of liver injury, and in the future, treatment of liver injury is expected to promote liver regeneration and to develop liver fibrosis inhibitor.

[0005]

An object of the present invention is to develop a therapeutic agent for hepatitis which can improve the liver function and can be administered orally or intravenously.

[0006]

Means for Solving the Problems The present inventors administered various amino acids to improve the pathology of galactosamine-administered rats used as a model for viral hepatitis or fulminant hepatitis, and found that alanine, glutamine and ornithine were the most excellent. The effect was obtained. That is, the present invention
A therapeutic agent for hepatitis comprising at least one of alanine, glutamine and ornithine.

[0007] It is known that the liver mitochondrial function is reduced during liver failure, and the redox potential of the liver and peripheral blood shifts to the reduced form due to suppression of the TCA cycle or β-oxidation of fat. That is, it is known that the ketone body ratio (acetoacetic acid / β-hydroxybutyric acid ratio), which reflects the redox potential of peripheral blood, is remarkably reduced when mitochondrial function is reduced, whereas alanine, glutamine and ornithine are The effect of suppressing the decrease in the peripheral blood ketone body ratio after the administration of galactosamine and maintaining the normal function of mitochondria was observed.

[0008] When various amino acids are administered after the induction of liver injury by the administration of galactosamine, the amino acids alanine, glutamine and ornithine improve the ODC activity, which is an index of liver regeneration, and have an effect of promoting liver regeneration. Plasma transaminase levels were markedly suppressed. From the above points, it has been clarified that these amino acids have a therapeutic effect that is not observed in conventional therapeutic agents for hepatitis. The therapeutic agent for hepatitis of the present invention is effective for acute or chronic, viral or drug-induced hepatitis, or fulminant hepatitis, and is also useful as a therapeutic agent for cirrhosis.

The therapeutic agent for hepatitis of the present invention contains one amino acid of alanine, glutamine and ornithine, and further effects can be obtained by containing these three amino acids in any combination. When two or three amino acids are contained, they may be contained as dipeptides or tripeptides.

[0010] The form in which the medicament of the present invention is provided includes:
For oral administration, for example, powders, granules, tablets, dragees,
For parenteral administration, for example, suspensions,
Detergents, emulsions, ampoules, injection solutions and the like can be mentioned, or a combination thereof can be provided. The injection may be in the form of an amino acid infusion and contains at least one of alanine, glutamine and ornithine, and the ratio of the total amount thereof to the total amino acid amount is 1%.
2% or more, or the daily dose of these amino acids
Preparations with 0 g or more can also be provided.

The form in which the food of the present invention is provided includes:
Powders, sugar-coated tablets, capsules, liquid preparations and the like can be mentioned.

The dose can be easily determined according to the symptoms, but the total amount of alanine and glutamine per adult per day is 1%.
g or more may be administered. Alanine and glutamine are each recognized as foods and are not likely to be toxic, especially acute.

[0013]

The present invention will be described more specifically with reference to the following examples.

[0014]

Example 1 Male SD rats (body weight 180-220)
g) was fasted for 5 hours, and 30% galactosamine hydrochloride (hereinafter abbreviated as Gal.N) saline solution was intraperitoneally administered (6.
1 hour before and 25 hours after administration of Gal · N, Ala, Gln, Orn, an amino acid mixture (casein composition except Ala, Gln) or an aqueous glucose solution was orally administered (2 g / kg BW),
This was compared with the case where only Gal · N was administered (group 1). 1 hour before Gal · N administration, 8, 24, 36 after administration
At 48 hours, blood was collected from the subclavian vein, and the ketone body ratio (plasma acetoacetic acid / β-hydroxybutyric acid ratio) was measured.

Table 1 below shows the treatments administered in each experimental group.
Table 2 shows the ketone body ratio after administration of Gal · N.

[0016]

[Table 1]

[0017]

[Table 2]

As shown in Table 2, no difference was observed between the groups up to 8 hours after administration of Gal · N, but at 24 hours, groups 1 (+ Gal · N) and 5 (Gal · N + amino acid mixture) ) And 6 groups (Gal · N + glucose), a decrease in the ketone body ratio was observed. Even in the 36th hour,
The decrease in ketone body ratio in groups 5 and 6 was remarkable, but 2
Group (Gal · N + Ala), 3 groups (Gal · N + Gl)
n) In group 4 (Gal · N + Orn), no decrease in ketone body ratio was observed, and almost normal values were maintained.

Although the peripheral blood ketone body ratio is known as an index reflecting the oxidation-reduction potential of the liver, the decrease in the ketone body ratio observed in the groups 1, 5, and 6 after administration of Gal · N is 2,
Ala, Gln was not observed in groups 3 and 4,
Alternatively, it is considered that mitochondrial function was normally maintained by administration of Orn.

[0020]

Example 2 SD male rats (body weight 180-220)
g) was fasted for 5 hours, and 30% galactosamine hydrochloride (G
al.N) saline solution intraperitoneally (600 mg /
1 kg before administration of Gal · N, Ala, Gl
n, Orn, amino acid mixture (casein composition but Al
a, except Gln) or an aqueous solution of glucose orally (2 g / kg BW), and compared with the case where only Gal · N was administered (group 1). Twenty-four hours after the administration of Gal · N, each animal was bled and killed, and the liver was excised, and then the hepatic ODC activity was measured.

Table 3 below shows the treatments administered in each experimental group.
FIG. 1 shows hepatic ODC activity.

[0022]

[Table 3]

The hepatic ODC activity was compared with that of one group (control group).
It was induced in group 2 (Ala), group 3 (Gln), and group 4 (Orn), but was more suppressed in group 5 (mixture of amino acids) and group 6 (glucose) than in the control. Ala, Gl
There has been no report that hepatic ODC activity was induced by administering n or Orn to a living body, and it was shown that these amino acids are effective for liver regeneration.

[0024]

Example 3 SD male rats (weight: 280-320)
g) was fasted for 5 hours, and 30% galactosamine hydrochloride (G
al.N) saline solution intraperitoneally (600 mg /
1% before and 25 hours after administration of Gal · N, 20% Ala, Gln (1: 1 mixture, 2 g / kg)
BW, 4 groups) or oral glucose solution (2
g / kg body weight, 3 groups) and compared with the case where only Gal · N was administered (2 groups). As a control, Gal
-The N non-administration group (normal animal) was also provided. Gal ・ N administration 4
Blood was collected from the subclavian vein 8 hours before and at 12, 24 and 36 hours after administration, and plasma GOT and GPT concentrations were measured. Ga
The animals were fasted from 44 hours after the administration of 1N, and the animals were bled and killed at 48 hours.

Table 4 below shows the administration treatment in each experimental group, Table 5 shows blood biochemical indices, and Table 6 shows.

The plasma cholesterol, triglyceride, phospholipid, total protein, and albumin concentrations were significantly reduced in the two groups (Gal.N only group) and three groups (Gal.N, Glucose group). However, such a decrease was not observed in the four groups (Gal · N, Ala + Gln administration group), and the plasma concentration was maintained at substantially the same level as in the Gal · N non-administration group.

According to the results of histopathological examination of the liver,
In the group, sporadic necrosis due to administration of Gal · N was observed, but in group 4, the occurrence of sporadic necrosis was suppressed, and Gal · N was suppressed.
Despite the administration of N, two cases were normal.

As shown in FIGS. 2 and 3, in groups 2 and 3 to which Ala and Gln were not administered, plasma GOT and GP were used.
Although the T concentration increased remarkably, the increase in plasma GOT and GPT concentrations was suppressed in the four groups to which Ala and Gln were administered. The plasma bilirubin concentration at the time of blood sampling and sacrifice also markedly increased by Gal administration in groups 2 and 3, but the increase in plasma bilirubin concentration was suppressed in group 4 to which Ala and Gln were administered. (FIG. 4)

[0029]

Example 4 SD male rats (body weight 180-220)
g) was fasted for 5 hours, and 30% galactosamine hydrochloride (G
al.N) saline solution intraperitoneally (600 mg /
kg) and 1 hour before administration of GalN
ucose, Ala, Ala + Gln, Gln (2 g each)
/ Kg BW) or a 20% aqueous solution of glutathione (50 mg / kg body weight) orally.
This was compared with the case where only N was administered (2 groups). Also,
As a control, a group without Gal · N administration (normal animals) was also provided. Blood was collected from the subclavian vein 48 hours before Gal · N administration and 12 hours after administration, and plasma GOT and GPT concentrations were measured. After fasting the animals 20 hours after administration of Gal · N,
Animals were bled and killed at 24 hours.

Table 7 below shows the administration treatment in each group, and Table 8 shows blood biochemical indices.

[0031]

[Table 7]

[0032]

[Table 8]

As shown in FIGS. 5 and 6, the plasma GOT and GPT concentrations in the Gal.N only administration group (group 2), the glucose administration group (group 3) and the glutathione administration group (group 7) were remarkable. Ala + Gln, Ala, Gln
In the group administered with, the increase in plasma GOT and GPT concentrations was suppressed, and the group administered with Ala + Gln was suppressed to the lowest level. The plasma bilirubin concentration was significantly increased by Gal administration in groups 2 and 3, but the increase in plasma bilirubin concentration was suppressed in group 4 to which Ala and Gln were administered.
(FIG. 7)

[0034] Plasma triglycerides, phospholipids, total protein,
Albumin concentration was 2 groups, Gal · N only administration group and 3 groups G
In the lucose-administered group, the decrease in plasma concentration due to liver injury was remarkable, but in the group Ala + Gln-administered group 5, the decrease in plasma lipid / protein concentration was the least. From the above results, the effects of the administration of Ala and Gln indicate that
Excellent effects were obtained as compared with the case of administration of Gln or Gln alone, and good results were obtained as compared with the existing drug, glutathione.

[0035]

Example 5 Male SD rats (body weight 180-220)
g) was fasted for 5 hours, and 30% galactosamine hydrochloride (G
al.N) saline solution intraperitoneally (600 mg /
kg) and 1 hour before administration of GalN
a + Gln, casein composition amino acid mixture, branched chain amino acid mixture (Val: Leu: Ile = 1: 2: 1.
2) Oral administration of Arg aqueous solution (2 g / kg body weight)
However, comparison was made with the case where only Gal · N was administered (2 groups). As a control, a group without Gal · N administration (normal animals) was also provided. 48 hours before Gal · N administration, 12 hours after administration
At the time, blood was collected from the subclavian vein and the plasma GOT and GPT concentrations were measured. The animals were fasted from 20 hours after administration of Gal · N, and then blood samples were killed 24 hours later.

Table 9 below shows administration treatments in each experimental group, and Table 10 shows blood biochemical indices.

[0037]

[Table 9]

[0038]

[Table 10]

As shown in FIGS. 8 and 9, the group administered with Gal · N alone (group 2), the group administered with the amino acid mixture (group 4), the group administered with the branched chain amino acid mixture (group 5), and the group administered with Arg (group 6) )), The plasma GOT and GPT concentrations were significantly increased, but in the Ala + Gln administration group (group 3),
The increase in GPT concentration was suppressed. Also in the plasma bilirubin concentration, the Ala + Gln administration group was suppressed to the lowest level,
In groups 2, 4, 5, and 6, there was a marked increase. (FIG. 10)

In this experiment, the degree of Gal liver injury was relatively mild as compared with the previous experiments. Therefore, there was no significant difference between the groups in the plasma cholesterol, phospholipid, total protein, and albumin concentrations, but no decrease in the plasma triglyceride concentration was observed in the groups 2, 4, 5, and 6. However, the three groups maintained the same level as the non-administration group. Although branched-chain amino acids and Arg are amino acids used for treatment of cirrhosis and liver failure, respectively.
In the case of Gal / N hepatic disorder that causes lesions similar to fulminant hepatitis, the effects of administration of branched-chain amino acids or Arg were not observed. In the amino acid mixture administration group, Ala,
The same effect as in the Gln group was not observed.

[0041]

Example 6 Male SD rats (body weight 180-220)
g) was fasted for 5 hours, and 30% galactosamine hydrochloride (G
al.N) saline solution intraperitoneally (600 mg /
kg) and 1 hour before administration of GalN
a + Gln is 0.5, 0.75, 1, 2, 3 g / kg
Was orally administered and only Gal · N was administered (2
Group). As a control, a group without Gal · N administration (normal animals) was also provided. 48 hours before Gal / N administration,
Blood was collected from the subclavian vein 12 hours after administration, and plasma GO was collected.
The T and GPT concentrations were measured. The animals were fasted from 20 hours after administration of Gal · N, and then blood samples were killed 24 hours later.

Table 11 shows the administration treatment in each experimental group.
Table 12 shows the blood biochemical indexes.

[0043]

[Table 11]

[0044]

[Table 12]

As shown in FIGS. 11 and 12, the plasma transaminase concentration at 24 hours after administration of Gal · N was Ala,
The increase in plasma concentration was suppressed depending on the dose of Gln.
In particular, in the groups administered with Ala and Gln at a dose of 2, 3 g / kg, a marked effect of suppressing the increase in plasma GOT and GPT concentrations was observed.
Plasma bilirubin concentration also depends on the dose of Ala, Gln,
The rise in plasma concentration was suppressed. (FIG. 13)

In two groups (Gal.N administration group), one group of Ga
Compared with the l · N non-administration group, the plasma cholesterol, triglyceride, phospholipid, and total protein concentrations, which are considered to be caused by liver damage, were reduced, but the reduction was improved according to the dose of Ala, Gln. A tendency was observed, and the values of the seven groups were almost the same as those of the first group.

[0047]

Example 7 Male SD rats (BW; 170-180)
g), after fasting for about 4 hours, Ala, Ala + Gln, O
0.3% of rn, Ala + Orn (2 g / kg BW each)
A carboxymethylcellulose (CMC) aqueous solution and an equivalent amount of a CMC aqueous solution alone were orally administered as a control, and one hour later, a saline solution of Gal · N (600 mg / kg BW) was intraperitoneally administered. Blood was collected 10 hours after administration, and GOT and GPT activities in plasma were measured. Twenty-four hours after administration, blood was collected and sacrificed, and blood biochemical indices were measured and liver histopathology was examined.

Table 13 below shows the treatments administered in each experimental group.

[0049]

[Table 13]

As compared to the Gal.N administration group, the four groups other than the Gal.N non-administration group tended to suppress the increase in plasma transaminase concentration. The effect of this suppression is
n, Ala + Orn, Ala + Gln, and Ala groups. From this result,
Orn was found to have an equal or greater effect of improving liver damage as compared to Ala and Gln.

[0051]

Example 8 Male SD rats (BW; 280-320)
g), Gal.N (500 mg / kg) after fasting for about 5 hours
BW) saline solution was injected from a pre-attached external jugular vein cannula. From 12 hours after Gal administration,
Peptide Ala · Gln saline solution (6 g / kgB
W) As a control, an equal volume of a physiological saline solution alone was injected from an external jugular vein catheter. 12, 24, 3 after Gal administration
Blood was collected at 4 hours, and GOT and GPT activities in plasma were measured. At 48 hours after administration, blood was collected and sacrificed, and blood biochemical indices were measured and liver histopathological examination was performed. (FIG. 16)

Even when Ala.Gln as a peptide was administered intravenously, a remarkable effect of suppressing an increase in plasma GOT concentration was observed, similarly to the case where a mixture of Ala and Gln was administered orally.

[0053]

According to the present invention, it is possible to provide a drug which improves liver function and has an excellent therapeutic effect on hepatitis, which is extremely useful in the pharmaceutical industry.

[Brief description of the drawings]

FIG. 1 is a diagram showing hepatic ODC activity in Example 2.

FIG. 2 is a view showing a plasma GOT concentration in Example 3.

FIG. 3 is a view showing a plasma GPT concentration in Example 3.

FIG. 4 is a graph showing the concentration of plasma bilirubin in Example 3.

FIG. 5 is a view showing a plasma GOT concentration in Example 4.

FIG. 6 is a graph showing the plasma GPT concentration in Example 4.

FIG. 7 is a graph showing the concentration of plasma bilirubin in Example 4.

FIG. 8 is a view showing a plasma GOT concentration in Example 5.

FIG. 9 is a graph showing the plasma GPT concentration in Example 5.

FIG. 10 is a graph showing the concentration of plasma bilirubin in Example 5.

FIG. 11 is a view showing a plasma GOT concentration in Example 6.

FIG. 12 is a graph showing the plasma GPT concentration in Example 6.

FIG. 13 is a graph showing the concentration of plasma bilirubin in Example 6.

FIG. 14 is a view showing a plasma GOT concentration in Example 7.

FIG. 15 is a view showing a plasma GPT concentration in Example 7.

FIG. 16 is a diagram showing a plasma GOT in Example 8.

[Table 4]

[Table 5]

[Table 6]

──────────────────────────────────────────────────続 き Continuation of the front page (56) References US Pat. No. 4,987,123 (US, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) A61K 31/198

Claims (3)

(57) [Claims] 1. A therapeutic agent for acute or chronic viral hepatitis, comprising at least one of alanine, glutamine and ornithine. 2. The therapeutic agent for acute or chronic viral hepatitis according to claim 1, which is any of alanine, glutamine, ornithine, alanine and glutamine, alanine and ornithine, or alanylglutamine. 3. The therapeutic agent for acute or chronic viral hepatitis according to claim 1, which is any of alanine, glutamine, ornithine, alanine and ornithine, or alanylglutamine.