RU2292875C2 - Quinone therapeutic agent for treatment of liver disease - Google Patents

Abstract

FIELD: medicine, oncology, pharmacy.

SUBSTANCE: invention describes a medicament used in treatment of liver disease by prevention of invasion onset of portal vein. This agent represents the effective medicament containing menatetrenone as an active component used in treatment/prophylaxis of liver disease. Indicated agent used in treatment/prophylaxis of liver disease is the effective medicament against liver cancer, in particular, against DCP (des-γ-carboxyprothrombin)-positive liver cancer. Agent used in aims for treatment/prophylaxis of liver disease contains menatetrenone as an active component, it improves prognosis significantly after carrying out the anticancer therapy and possesses the excellent effect directed on prevention of liver cancer relapses.

EFFECT: valuable medicinal property of agent.

4 cl, 2 tbl, 10 dwg, 2 ex

Description

FIELD OF THE INVENTION

The present invention relates to an agent for the treatment of liver diseases, and more particularly to an agent that improves the prognosis of liver cancer and contains menatetrenone as an active ingredient.

State of the art

It is known that patients with hepatocellular carcinoma (hereinafter referred to as “HCC”) have a high level of portal vein invasion (hereinafter referred to as “IVV”), and after IVV the prognosis becomes very poor. It is known that a high level of des-γ-carboxyprothrombin (hereinafter referred to as “DCP”) in patients with HCC is closely associated with the subsequent development of TRS (see Koike Y., Cancer 2001, 91: 561-9). DCP is also referred to as PIVKA-II (a protein induced by a lack of vitamin K or an antagonist II). DCP is prothrombin, which does not have normal coagulating activity, and it is known that in the event of a deficiency of vitamin K (hereinafter referred to as “VK”), its level increases; thus, DCP is a protein used as a marker for VK deficiency or impaired VK absorption.

In addition, it was reported that after administration of VK to patients with HCC having a high level of DCP, serum DCP decreased (see Cancer 1992, 69: 31-8), and after administration of vitamin K-II (hereinafter referred to as “VK-II” ") in DCP-producing fcc cell lines in vitro , inhibition of cell proliferation was observed (see Hepatology, 1995, 22: 876-82).

However, until now, no clinical data have been obtained indicating the possibility of preventing IVV and relapse of hepatocellular carcinoma by administering VK-II to patients after a course of HCC treatment, where such an introduction would lead to an improvement in the prognosis of this disease.

In accordance with the above, an object of the present invention is to provide a highly effective agent for treating or preventing liver diseases.

Description of the invention

The basis of the present invention was mainly the authors' discovery of the fact that oral administration of VK-II to patients suffering from HCC with the production of DCP leads to inhibition of IVV after treatment with HCC and to an improvement in the prognosis of this disease, as well as to suppress relapse liver cancer after treatment.

The above goal is achieved by using an agent for the treatment or prevention of liver diseases containing menatetrenone as an active ingredient.

In accordance with a preferred aspect of the present invention, the aforementioned agent is used when said liver disease is liver cancer.

In accordance with a preferred aspect of the present invention, the aforementioned agent is used when said liver cancer is a liver cancer in which des-γ-carboxyprothrombin (DCP) is produced.

In accordance with a preferred aspect of the present invention, the above agent improves the prognosis of liver cancer after treatment.

In accordance with a preferred aspect of the present invention, the aforementioned agent is an agent that prevents portal vein invasion (IVV).

In addition, the aforementioned goal is achieved using the means for preventing portal vein invasion (IVV), wherein said agent contains menatetrenone as an active ingredient.

In addition, the aforementioned goal is achieved using an agent for increasing patient survival after treatment for liver cancer, wherein said agent contains menatetrenone as an active ingredient.

In addition, the above objective is achieved using an agent for preventing relapse of hepatocellular carcinoma containing menatetrenone as an active ingredient.

In addition, the above objective is achieved by using a DCP-lowering agent and containing menatetrenone as an active ingredient.

In addition, the above goal is achieved by applying a method for preventing portal vein invasion (IVV), comprising administering to the patient an effective dose of a medicament containing menatetrenone as an active ingredient.

In addition, the above objective is achieved by applying a method for preventing relapse of hepatocellular carcinoma, comprising administering to the patient an effective dose of a medicament containing menatetrenone as an active ingredient.

In addition, the above goal is achieved by applying a method of regulating the level of DCP in the blood of a patient, comprising administering to the patient an effective dose of a medicament containing menatetrenone as an active ingredient.

In addition, the above goal is achieved using menatetrenone in order to obtain funds to prevent IVV.

In addition, the above goal is achieved using menatetrenone in order to prevent recurrence of hepatocellular carcinoma.

In addition, the above goal is achieved using an agent for treating or preventing liver disease, wherein said agent contains vitamin K as an active ingredient.

An agent containing menatetrenone for the treatment of liver diseases in accordance with the present invention has a highly effective effect, preventing IVV in liver disease, and in particular, DCP-positive liver cancer, and, in addition, has a highly effective effect improving the prognosis after treatment of liver cancer. In addition, an agent containing menatetrenone for treating liver diseases in accordance with the present invention is most suitable for preventing liver cancer after treatment.

A brief description of the graphic material

Figure 1 presents a block diagram illustrating the selection of patients.

2 is a graph illustrating a change in serum DCP level.

Figure 3 presents a graph illustrating the change in the frequency of cases of TRS.

4 is a graph illustrating changes in survival.

Fig. 5 is a graph illustrating the inhibitory effect of VK-II administration to prevent relapse of liver cancer (50% of relapses).

6 is a graph illustrating the results only in the case of HCV infections in tests conducted to confirm the inhibitory effect of the introduction of VK-II, aimed at preventing relapse of liver cancer (50% relapse).

7 is a graph illustrating the results in the case of eliminating the occurrence of local relapses in tests conducted to confirm the inhibitory effect of the introduction of VK-II, aimed at preventing relapse of liver cancer (50% of relapses).

On Fig presents a graph illustrating the results in the case of eliminating the occurrence of relapse within 6 months in trials conducted in order to confirm the inhibitory effect of the introduction of VK-II, aimed at preventing relapse of liver cancer (50% of relapses).

9 is a diagram illustrating the results of the assessment of the relative risk (RR) of relapse using the Cox proportional risk model and

Figure 10 presents a graph illustrating the results of analyzes on the level of DCP before treatment and after relapse.

BEST MODES FOR CARRYING OUT THE INVENTION

The following is a more detailed description of the present invention with reference to examples, which, however, should not be construed as limiting the invention.

Liver cancer often occurs in patients with chronic hepatitis and cirrhosis of the liver, the treatment of which the present invention is directed, and after courses of such treatment its relapses are very often observed. So, for example, type C hepatitis or type B hepatitis leads to cirrhosis of the liver, and relapses occur after excision of the tumors. Using the agent of the present invention for the treatment of liver disease, the prognosis after such treatment for liver cancer can be significantly improved (i.e., relapses can be prevented or removed). In addition, with the help of this tool can be successfully prevented the invasion of the portal vein (IVV), which is a form of relapse of liver cancer with a poor prognosis.

Menatetrenone used in the present invention has the chemical name 2-methyl-3-tetraprenyl-1,4-naphthoquinone, and its structural formula is presented below.

Menatetrenone is a yellow crystalline or oily substance that has neither taste nor smell and is easily decomposed by light. In addition, menatetrenone is almost insoluble in water. Menatetrenone is also called vitamin K-II (VK-II), and with regard to its pharmacological action, then in the process of protein synthesis of blood coagulation factors (prothrombin VII, IX and X), menatetrenone is involved in the carboxylation reaction, where glutamic acid residues turn into physiologically active γ-carboxyglutamic acid, and also stimulates the synthesis of normal prothrombin in the liver, etc. and activates the mechanism of hemostasis in a living organism, that is, contributes to the implementation of hemostasis in the body.

Menatetrenone, which is the active ingredient in the drug of the present invention, may be present in the form of an anhydride or hydrate. In addition, menatetrenone has crystalline polymorphs, but is not limited to them; however, menatetrenone may be present in any crystalline forms, which are, but are not limited to, crystalline forms or mixtures. In addition, the metabolite produced by menatetrenone of the present invention and decomposable in a living organism is also included in the scope of the claims of the present invention.

Menatetrenone used in the present invention can be easily produced by a well-known method, and a representative example of a method for its preparation is the method described in Japanese Patent Application Laid-Open No. 49-55650; or, alternatively, such menatetrenone may be purchased from chemical manufacturers. In addition, menatetrenone can be purchased as a capsule or injection drug. As for the drug of the present invention, menatetrenone can be used in pure form, or it can be prepared by the standard method in the form of a pharmaceutical preparation by mixing with ingredients that are commonly used as starting materials for the preparation of drugs, such as well-known pharmaceutically acceptable carriers, etc. (e.g. fillers, binders, disintegrants, lubricants, dyes, flavoring agents, stabilizers, emulsifiers, absorption stimulants, surfactants, pH regulators, preservatives, antioxidants, etc.). In addition, if necessary, other ingredients such as vitamins and amino acids can be mixed. Examples of pharmaceutical formulations are tablets, powders, granules, capsules, syrups, suppositories, injectable preparations, ointments, poultices, and the like.

In addition, according to the present invention, the administration method of menatetrenone is not particularly limited, although oral administration is preferred. At the same time, male-containing capsules can be purchased in the form of capsules under the name “Kaytwo capsules” (the patented name given by Eisai Co., Ltd.) and in the form of capsules under the name “Glakay capsules” (the patented name given by Eisai Co., Ltd.) , a methethenone-containing syrup can be purchased in the form of a syrup called "Kaytwo syrup" (the proprietary name given to Eisai Co., Ltd.), and an injection drug can be purchased in the form of a drug called "Kaytwo N" (the patented name given by Eisai Co., Ltd.).

The menatetrenone-containing drug of the present invention can be used to treat or prevent liver disease. A preferred dose of menatetrenone is generally 10-200 mg / day, more preferably 30-135 mg / day.

Examples

The following are examples of tests of the present invention, however, these examples are for illustrative purposes only and should not be construed as limiting the present invention. Each specialist can implement the present invention not only in accordance with the following test examples, but he can also make various modifications without departing from the scope of the claims, and such modifications are also included in the scope of the claims of the present invention.

Test Example I

Clinical trials (randomized prospective controlled trials) were performed as follows.

Patients with hepatocellular carcinoma with a DCP level above 60 IU / L (DCP-positive liver cancer) were included in this trial. On the other hand, patients with portal vein invasion and patients who already had VK metabolism as a result of VK administration or a VK antagonist were excluded from this trial. A detailed description of the tests is given in table 1.

Table 1

Individuals participating in the trial:

The following individuals were included in the trial:

1. Patients with liver cancer.

2. Patients with a serum DCP level of ≥ 60 IU / L.

The following individuals were excluded from the tests:

1. Individuals with invasion of the portal vein.

2. Individuals with metastases of liver cancer to peripheral organs.

3. Individuals with unregulated ascites production.

4. Individuals with bilirubin levels> 3.0 mg / dl.

5. Individuals taking the vitamin K drug, warfarin.

Groups administered by VK-II:

Patients who took 45 mg of vitamin K-II (Glakay) three times after liver cancer treatment.

Groups not administered K-II:

Patients who have undergone only liver cancer treatment.

Criteria for evaluation:

1. The occurrence of portal vein invasion.

2. Death.

Figure 1 presents a block diagram illustrating the selection of patients. 126 patients with liver cancer were treated from February 1999 to November 2001. Treatment of liver cancer consisted of therapy with HCC by percutaneous cauterization (RFA and / or PEIT); treatment through blood vessels (TAE or TAI); or in a surgical operation. 5 patients were excluded from this trial.

Then the rest (121 patients) were randomly divided into a group of patients who took VK-II (n = 60) and a group of patients (control group) who did not take VK-II (n = 61). The group that took VK-II received orally 45 mg / day of VK-II (the proprietary name Glakay given by Eisai Co., Ltd.) after treatment for liver cancer, and the control group did not receive VK-II.

After liver cancer treatment, patients were monitored. This observation of patients consisted of an ultrasound every 3 months (abdominal ultrasound imaging), computed tomography every 6 months and an assessment of the levels of tumor markers, alpha-fetoprotein and DCP, every month.

Table 1 presents the clinical data for the studied patients. There were no significant differences between the clinical parameters for the group taking the drug in question and for the group not taking such a drug.

Table 1
Description of clinical data for patients The group taking the drug in question
(n = 60) Group not taking the drug in question
(n = 61) P Age 66.9 ± 7.0 67.3 ± 7.5 0.8 Gender (male / female) 36/24 45/16 0.12 Virus (HCV / non HCV) 50/10 52/9 0.81 Tumor Size (mm) 32 ± 11 35 ± 18 0.27 The number of tumors 4.0 ± 3.2 4.3 ± 3.5 0.66 Infantile hepatitis, class (A / B or C) 18/42 27/34 0.13 Albumin (g / dl) 3.4 ± 0.5 3.5 ± 0.5 0.3 Bilirubin (mg / dl) 1.2 ± 0.7 1.1 ± 0.9 0.4 ALT (ME / L) 55 ± 38 61 ± 47 0.47 Prothrombin (%) 78 ± 16 78 ± 14 0.99 Blood platelets (10 ⁴ / mm ³ ) 10.8 ± 6.0 11.5 ± 6.6 0.52 AFP (ng / l) 2668 ± 7666 1539 ± 7036 0.42 DCP (ME / L) 985 ± 2639 1178 ± 5108 0.80 PTA with / without drug administration 48/12 41/20 0.15 mean ± cf sq. from. (median)

2 is a graph illustrating changes in serum DCP levels. The solid line indicates the group that has taken the drug, and the dotted line means the group that has not taken the drug. For both groups, after treatment for liver cancer, the level of DCP after decreased. Then, the DCP level in the group taking the drug remained approximately constant for 12 months, while the DCP level in the group not taking this drug gradually increased.

Figure 3 presents a graph illustrating changes in the frequency of cases of TRS. As shown in figure 3, in the group taking the drug in question, the number of cases of TRS was 2% after 1 year, and 23% after 2 years. On the other hand, in the group taking the drug in question, the number of cases of IVS was 23% after 1 year, and 47% after 2 years (P = 0.018).

4 is a graph illustrating a change in survival. As shown in figure 4, the survival rate of patients in the group taking the drug in question was 66% after 2 years, while the survival rate of patients in the group not taking this drug was 28% (P = 0.044).

A statistical analysis was conducted on the incidence of IVV and the survival of patients in these two groups. That is, the calculations were performed using the Cox proportional risk model, and this test was performed using the log-rank criterion. The mean follow-up period was 12 ± 8 months.

Based on the above results, it was hypothesized that with the oral administration of VK-II, there was a very sharp decrease in the number of cases of IVS in patients with DCP-positive HCC, and, in addition, survival in these patients increased significantly, and therefore significantly improved prognosis after treatment of liver cancer.

Test Example 2

The test was carried out as described below, in order to study the inhibitory effect of VK-II, aimed at preventing the recurrence of hepatocellular carcinoma after its treatment, and its safety.

For the period from March 1999 to March 2001, a list was compiled, which included 61 patients with diagnosed hepatocellular carcinoma and subsequent complete necrosis after treatment (or surgery), as indicated by contrast computed tomography; and the patients who were included in this list were divided into two groups, namely, the group that took VK-II and in which the final number of patient IDs was odd, and the group that did not receive VK-II (control group) and in which the final number of patient IDs was even; while patients of the VK-II group were orally administered VK-II (the proprietary name Glakay, given by Eisai Co., Ltd.) at a dose of 45 mg / day. Then, every 3 months, a contrast computed tomography or NMR tomography was performed, as well as a statistical analysis of the period of time until relapse occurred. In particular, a comparison was made using the Kaplan-Mayer criterion (log-rank test) and an assessment of the relative risk of relapses using the Cox proportional risk model.

As shown in Table 2, the mean follow-up period for 61 trial participants (32 patients in the VK-II group and 29 patients in the VK-II group) was 19.6 months (7-32).

table 2 Individuals Group hosted VK-II (32 cases) Control group (29 cases) Age 63.3 ± 7.5 (48-75) 64.5 ± 67 (45-74) Gender (male / female) 23/9 18/11 Disease causing factor (type C / type B / type B + C) 28/3/1 26/2/1 Background to alcohol use (alcoholic / non-alcoholic) 10/22 3/26 Primary swelling / relapse 15/17 14/15 Tumor Size (mm) 17.7 ± 5.1 (10-30) 19.4 ± 6.9 (10-38) The number of tumors 1.50 ± 0.88 (1-4) 1.48 ± 0.74 (1-3) Log AFP (ng / ml) 1.47 ± 0.61
(0.60-3.09) 1.72 ± 0.91
(0.48-3.88) PIVKA-II (mAE / ml) 41.8 ± 65.4
(8-346) 70.3 ± 104.1
(7-417) Liver Function (LD A / B / C) 15/16/1 13/15/1 Method of treatment (surgical / non-surgical) 1/31 3/26 Average follow-up period (months) 24.3 ± 7.1 (13-37) 24.2 ± 8.3 (12-37)

After calculating the total number of cases of relapses, it was found that the number of relapses 1 year after treatment was 12.5% for patients in the group receiving VK-II, and 55.2% for patients in the control group, while the number of relapses after 2 years after treatment, the patients in the group taking VK-II amounted to 39.6%, and in patients in the control group - 85.5%. Based on these results, it was found that, compared with the control group, the group taking VK-II showed a statistically significant decrease in the total number of cases of liver cancer recurrence.

Fig. 5 is a graph illustrating the effect of VK-II administration aimed at preventing relapse of liver cancer (50% of relapses). As shown in FIG. 5, the period before the onset of liver cancer relapses (50% of relapses) in the group that took VK-II was 26 months, and in the control group it was 10 months.

Moreover, after calculating the total number of relapse cases, taking into account only HCV cases (hepatitis C cases), it was found that the occurrence of relapse 1 year after treatment in patients treated with VK-II was 7.1%, and in patients with control group - 61.5%, and the occurrence of relapse 2 years after treatment in patients of the group taking VK-II was 37.8%, and in patients of the control group - 87.2%. Based on these results, it was found that, compared with the control group, the group that took VK-II showed a statistically significant decrease in the overall frequency of liver cancer recurrences when considering only HCV cases.

6 is a graph illustrating the results obtained in tests only for HCV cases conducted in order to confirm the inhibitory effect of the introduction of VK-II, aimed at preventing relapse of liver cancer (50% relapse). As shown in FIG. 6, the period before the onset of relapse of liver cancer (50% of relapses) in the group that took VK-II was 26 months, and in the control group it was 10 months.

Fig. 9 is a diagram illustrating the results of assessing the relative risk (RR) of relapse using the Cox proportional risk model. As shown in Fig. 9, if we take the relative risk for the control group as 1, then the relative risk for the group taking VK-II was approximately one third, that is, 0.329; in particular, for HCV cases, the relative risk was 0.210, that is, for the group taking VK-II, this risk was reduced by five times.

7 is a graph illustrating the results for cases of exclusion of local relapses in trials to confirm the inhibitory effect of VK-II administration aimed at preventing liver cancer recurrence (50% relapse) (VK-II group: 29 patients; group not taking VK-II: 22 patients). In addition, Fig. 8 is a graph illustrating the results in the case of eliminating the occurrence of relapse within 6 months in tests conducted to confirm the inhibitory effect of the introduction of VK-II, aimed at preventing relapse of liver cancer (50% of relapses) (group that took VK-II: 31 patients; group not taking VK-II: 22 patients). As shown in FIGS. 7 and 8, in these cases it was again established that the group that took VK-II showed a statistically significant decrease in the overall frequency of liver cancer relapses compared with the control group.

Figure 10 presents a graph illustrating the results of analyzes on the level of DCP before treatment and after relapse. As shown in figure 10, in all cases of relapse in patients of the group receiving VK-II, DCP was not detected, and, in addition, there were no side effects and cases of dropping out of the tests.

The effect of VK-II of the present invention on the development of invasion / metastasis of liver cancer cells has been investigated in vitro . The invasive ability was investigated using an invasion assay using HepG2 cells and a Matrigel chamber. The results showed that the number of cells passing through the Matrigel chamber decreased with the addition of VK-II, and this decrease depended on the concentration of VK-II. The ability to act on metastases was studied by the effect of VK-II on the expression of intercellular matrix metalloproteinases (MMPs) by Western blot analysis. After testing for the expression of MMP-1 and MMP-3 proteins when VK-II was added to liver cancer cells, it was found that expression was inhibited in this case. Based on these results, although they were obtained in vitro , it can be assumed that VK-II inhibits the invasion / metastasis of liver cancer cells.

Industrial application

As described above, the menatetrain-containing agent used to treat or prevent liver diseases in accordance with the present invention has an excellent inhibitory effect on TRS in liver disease, and in particular, on DCP-positive liver cancer, and, in addition, it has an excellent effect. in terms of prognosis after treatment of liver cancer.

In addition, the menatetrain-containing agent used to treat or prevent liver diseases in accordance with the present invention is very effective in preventing recurrence of liver cancer after treatment.

Claims (4)

1. An agent for inhibiting the recurrence of hepatocellular carcinoma after treatment of liver cancer, containing menatetrenone as an active ingredient. 2. An agent that reduces the level of des-γ-carboxyprothrombin and contains menatetrenone as the active ingredient. 3. A method of inhibiting the recurrence of hepatocellular carcinoma, comprising administering to the patient after treatment for liver cancer, an effective dose of a medicament containing menatetrenone as an active ingredient. 4. A method for modulating the level of des-γ-carboxyprothrombin in the blood of a patient, comprising administering to said patient an effective dose of a medicament containing menatetrenone as an active ingredient.

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