Pharmaceutical compositions for the treatment or prevention of Epstein-Barr virus hepatitis

Abstract

To provide novel agents for the treatment or prevention of Epstein-Barr virus hepatitis. [Solution] A pharmaceutical composition for the treatment or prevention of Epstein-Barr virus hepatitis in a subject, comprising ibudilast, dipyridamole, or both as active ingredients.

Description

[Technical Field]

[0001] This disclosure relates to pharmaceutical compositions for the treatment or prevention of Epstein-Barr virus (EBV) hepatitis. [Background technology]

[0002] Viral hepatitis is an inflammatory disease of the liver caused by infection with hepatitis viruses. Viral hepatitis is classified into different types depending on the type of causative virus. The main types are hepatitis A, B, C, and E, but it is estimated that so-called non-ABCE hepatitis accounts for approximately one-third of all viral hepatitis cases. In Japan, the annual number of deaths from non-ABCE viral hepatitis can reach approximately 100 per 100,000 people.

[0003] One of the major non-ABCE viral hepatitis types is Epstein-Barr (EB) virus hepatitis. Epstein-Barr virus (EBV) is a virus belonging to the Herpesviridae family, Gammaherpesvirinae subfamily. EBV infects humans and is associated with a variety of diseases, but its infection process involves multiple stages. These stages include "primary infection," "latent infection" (where the virus remains dormant in the host after primary infection, expressing only a portion of its genes without necessarily causing disease), and "reactivation" (which is triggered by factors such as a weakened immune system in the host). EBV infection itself is not uncommon; in fact, most adults have a latent EBV infection. EBV is known to be transmitted from person to person through saliva. Primary infection often occurs in infancy and is usually asymptomatic, but sometimes (especially if the primary infection occurs after puberty) it can cause infectious mononucleosis (IM). It is believed that there is an incubation period of approximately 4 to 6 weeks between actual viral infection and the onset of disease. Infectious mononucleosis is characterized by fever, redness of the throat, and enlargement of the spleen, and about 70% of patients also develop hepatitis, or EBV hepatitis. EBV hepatitis can be severe and accompanied by jaundice.

[0004] In contrast to ABCE viruses, which infect the liver, EBV does not infect liver cells, so the mechanism of EBV hepatitis development in the liver is unknown. However, it is thought to be a secondary outcome of viral infection and involves the host immune system in response to viral antigens (Non-Patent Literature 1). Non-Patent Literature 2 and 3, with the present inventor as the lead author, describe mouse experiments using MHV68, a mouse model of EBV that also causes IM-like symptoms, in which lymphocyte infiltration was observed in the liver tissue of virus-infected mice, and the levels of liver enzymes (serum concentrations of AST (aspartate aminotransferase) and ALT (alanine aminotransferase)), which are liver damage markers, increased. They also describe that the above lymphocyte infiltration and liver enzyme levels were suppressed by oral administration of intestinal non-penetrating antibiotics such as neomycin and vancomycin. The latter finding suggested that bacteria in the intestinal tract have some indirect influence on inflammation in the liver. Non-patent document 3 further describes that administration of anti-TLR4 (Toll-like receptor 4) antibodies, or the TLR4 inhibitor C34, or TLR4 knockout suppresses hepatitis in virus-infected mice, as indicated by the lymphocyte infiltration and hepatic enzyme levels described above. TLR4 is a receptor that recognizes lipopolysaccharide (LPS), a component of the outer membrane of Gram-negative bacteria, and induces an innate immune response. The disclosure in non-patent document 3 also describes hepatic enzyme levels and IFN-γ + Regarding the degree to which hepatitis can be suppressed, as expressed by the level of hepatic infiltration of CD8 cells and NK (natural killer) cells, there were no significant differences among anti-TLR4 antibodies, TLR4 inhibitor C34, and TLR4 knockout. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] American Journal of Pathology (2006) 168: 1057-1059. [Non-Patent Document 2] Journal of Immunology (2018) 200: 2703-2713. [Non-Patent Document 3] Journal of Immunology (2023) 211: 1550-1560. [Overview of the project]

[0006] This disclosure is based on the surprising finding that dipyridamole, known as an antiplatelet agent, and ibudilast, known as a bronchial asthma agent, can exhibit significantly superior antiviral hepatitis-suppressing effects compared to C34, and provides novel agents for the treatment or prevention of Epstein-Barr virus hepatitis.

[0007] More specifically, this disclosure includes the following embodiments. [1] A pharmaceutical composition comprising ibudilast, dipyridamole, or both, as active ingredients, for the treatment or prevention of Epstein-Barr virus hepatitis in a subject. [2] The pharmaceutical composition described above comprises ibudilast, as in [1]. [3] The pharmaceutical composition described above comprises dipyridamole, as described in [1]. [4] The subject is a subject having a primary infection with the Epstein-Barr virus, as described in any one of the claims [1] to [3]. [5] The subject is a subject having infectious mononucleosis, and the pharmaceutical composition is as described in any one of [1] to [4]. [Brief explanation of the drawing]

[0008] [Figure 1] Figure 1 shows (a) the change in mouse body weight over the number of days after viral inoculation, and (b) the amount of liver enzymes released, for experiments involving dipyridamole (DPM) administration. [Figure 2] Figure 2 shows (a) the change in mouse body weight over the number of days after viral inoculation, and (b) the amount of liver enzymes released, for experiments involving ibudilast (IBD) administration. [Figure 3] Figure 3 shows the antiviral hepatitis-suppressing effects of ibudilast and dipyridamole compared to C34. [Modes for carrying out the invention]

[0009] This disclosure provides pharmaceutical compositions for the treatment or prevention of EBV hepatitis in subjects. The pharmaceutical compositions comprise ibudilast, dipyridamole, or both as active ingredients. In some embodiments, the pharmaceutical composition comprises ibudilast. In some embodiments, the pharmaceutical composition comprises dipyridamole. In some embodiments, the pharmaceutical composition comprises both ibudilast and dipyridamole. Various embodiments of this disclosure may also be described as ibudilast and / or dipyridamole for the treatment or prevention of EBV hepatitis in subjects. Various embodiments of this disclosure may also be described as methods for the treatment or prevention of EBV hepatitis in subjects, comprising administering ibudilast and / or dipyridamole to subjects. Various embodiments of this disclosure may also be described as the use of ibudilast and / or dipyridamole in the manufacture of a drug for the treatment or prevention of EBV hepatitis.

[0010] Epstein-Barr virus (EBV) is a virus that infects humans and, like MHV68 which infects mice, belongs to the Herpesviridae family, Gammaherpesvirinae subfamily. EBV hepatitis is well known to those skilled in the art and is typically caused by primary EBV infection, particularly typically in association with infectious mononucleosis. Therefore, in some embodiments of this disclosure, the subjects are subjects having primary EBV infection. In some embodiments of this disclosure, the subjects are subjects having infectious mononucleosis. "Subjects" may also be called "patients". In some embodiments, the subjects are subjects who do not have one or more of the following conditions: bronchial asthma, history of cerebral infarction, history of stroke, cerebral ischemia, angina pectoris, myocardial infarction, ischemic heart disease, congestive heart failure, history of heart valve replacement, nephrotic syndrome, chronic glomerulonephritis, arteriosclerosis obliterans, peripheral artery disease, pulmonary hypertension, multiple sclerosis, and neuropathic pain.

[0011] Ibudilast, also known as AV-411 or MN-166, is approved in Japan for the treatment of bronchial asthma and dizziness caused by chronic cerebral circulatory disorders associated with post-stroke sequelae. While ibudilast's primary mechanism of action is phosphodiesterase inhibition, it is also known to potentially inhibit TLR4 (e.g., Transplant Proc. 2020, 52(6):1869-1874). Non-patent document 3 disclosed results suggesting a link between EBV hepatitis and TLR4. However, the results disclosed in non-patent document 3 indicated that anti-TLR4 antibodies, the TLR4 inhibitor C34, and TLR4 knockout suppressed hepatitis to a similar degree, and did not suggest, for example, that other TLR4 inhibitors could show a significantly superior hepatitis-suppressing effect compared to C34. Furthermore, ibudilast is known to have liver dysfunction accompanied by elevated AST, ALT, etc., as a major side effect (Ibudilast Capsules "Ketas Capsules 10mg" Drug Interview Form, Revised May 2024, Kyorin Pharmaceutical Co., Ltd.). While many compounds are known to those skilled in the art to exhibit TLR4 inhibitory activity, the fact that ibudilast showed a hepatitis-suppressing effect superior to that of C34 was an unexpected discovery.

[0012] Dipyridamole is a drug known as a coronary circulation improving agent and antiplatelet agent. Thome et al., Antiviral Research 172 (2019) 104615 reported that dipyridamole inhibited the reactivation of EBV from B cell lines with latent EBV infection. However, the fact that dipyridamole could suppress EBV hepatitis, which is understood to occur as a secondary consequence of primary EBV infection, was not necessarily a predictable finding.

[0013] As described in Non-Patent Documents 2 and 3, the suppression of EBV hepatitis can be observed and measured as a decrease in serum concentrations of hepatic enzymes such as AST and ALT. Alternatively, as also described in Non-Patent Documents 2 and 3, the suppression of EBV hepatitis can be measured as a decrease in IFN-γ infiltrating the liver. + CD8+ T cell count or IFN-γ + This can be observed and measured as a decrease in NK cell count. AST (aspartate aminotransferase) is sometimes called GOT (glutamate oxaloacetate transaminase). ALT (alanine aminotransferase) is sometimes called GPT (glutamate pyruvate transferase). If the pharmaceutical composition is administered after primary EBV infection but before these hepatitis conditions develop, it may prevent EBV hepatitis; if the pharmaceutical composition is administered after these hepatitis conditions have developed, it may treat EBV hepatitis. Administration of the pharmaceutical composition of the embodiment may result in serum concentrations of the above-mentioned liver enzymes and IFN-γ infiltration into the liver compared to when it is not administered or compared to before administration. + CD8 + T cell count and IFN-γ + This can lead to a decrease in the number of NK cells.

[0014] A pharmaceutical composition according to an embodiment of the present disclosure may include a pharmaceutically acceptable carrier or excipient in addition to an active ingredient. Therefore, the manufacture of the medicament of the embodiment may include combining ibudilast and / or dipyridamole with a pharmaceutically acceptable carrier or excipient. Specific examples of such carriers or excipients include water (generally, an aqueous solution as a liquid preparation may also contain other components such as buffers and inorganic salts as known to those skilled in the art), propylene glycol, ethanol, vegetable oil, mineral oil, dimethyl sulfoxide, glycerin, lactose, sucrose, mannitol, starch, dextrin, cellulose, cellulose ether, polyethylene glycol, polyvinyl pyrrolidone, magnesium stearate, talc, and any combination thereof, but are not limited thereto. The pharmaceutical composition according to an embodiment of the present disclosure can administer a therapeutically effective amount of ibudilast and / or dipyridamole in various dosage forms and administration routes to a subject who needs treatment or prevention of EBV hepatitis. Since ibudilast and dipyridamole are drugs that have been used as pharmaceuticals for different purposes, dosage forms and administration routes suitable for them are known to those skilled in the art. For example, the pharmaceutical composition can be administered in the form of an aqueous solution, tablet, pill, powder, granule, or capsule. Suitable administration routes include, but are not limited to, intraperitoneal, intravenous, and oral administration.

Example

[0015] Hereinafter, examples are shown to explain the embodiment in more detail. However, the examples are for illustrative purposes only, and the embodiments of the invention are not limited to these examples. In this example, experiments were conducted using the MHV68 infection system established as a mouse model of EBV infection. In the figure, * indicates statistical significance of P < 0.05.

[0016] All mice received 10 6Mice were infected by intratracheal inoculation with the pfu MHV68 virus. Dipyridamole (DPM) at 200 μg / kg was administered intraperitoneally daily to the DPM group, and the corresponding amount of solvent (0.8% DMSO in phosphate-buffered saline (PBS)) was administered intraperitoneally daily to the control group starting from the 4th day after virus inoculation. Figure 1a shows a plot of the change in mouse body weight over the number of days after virus inoculation. Each point represents the average value of three mice per group, and the error bars represent the standard deviation. Virus infection causes a rapid decrease in body weight starting around the 5th day, but administration of DPM can significantly suppress the weight loss.

[0017] Fourteen days after virus inoculation, sera were prepared from the peripheral blood of each mouse, and the amounts of hepatic escape enzymes AST and ALT were measured based on enzyme activity by a conventional method as described in Non-Patent Documents 2 and 3. The results shown in Figure 1b represent the average values of six mice each in the DPM group and the control (DMSO) group, and the error bars represent the standard deviation. Compared to the non-infected control (PBS), virus infection (MHV68) causes a significant increase in the amount of hepatic escape enzymes (the third from the left in each graph in Figure 1b), and + CD8 + a significant infiltration of CD8 T cells and IFN-γ + NK cells into the liver also occurs (not shown; see Non-Patent Documents 2 and 3). This corresponds to the state of EBV hepatitis in human infectious mononucleosis. It can be seen that viral hepatitis was significantly suppressed in the group that received DPM following virus infection.

[0018] Figure 2 shows an experiment of ibudilast (IBD) administration. Similar to the above, all mice were given 10 6PFU-mediated MHV68 virus was inoculated intratracheally to induce infection. The IBD group received 100 μg / kg of IBD, while the control group received a corresponding amount of solvent (0.8% DMSO in PBS), administered intraperitoneally daily starting 4 days after viral inoculation. Figure 2a plots the change in mouse body weight against the number of days after viral inoculation. Each point represents the mean value for 3 mice in each group, and the error bars represent the standard deviation. IBD showed a noticeable effect in suppressing weight loss, similar to, or even more significantly than, DPM described above. Serum levels of liver enzymes were also measured in the same manner as above (Figure 2b). As can be seen in this figure, IBD was able to demonstrate a dramatic suppressive effect against viral hepatitis.

[0019] Figure 3 shows a side-by-side comparison of the suppression of hepatic enzyme levels by IBD and DPM in virus-infected mice with the suppression of hepatic enzyme levels using C34 (a TLR4 inhibitor described in Non-Patent Literature 3). The procedures for viral infection, drug administration, and measurement of hepatic enzyme levels were as described above. The dosages of C34, IBD, and DPM were 1000 μg / kg, 100 μg / kg, and 200 μg / kg, respectively. It can be seen that IBD and DPM exerted significantly greater inhibitory effects on viral hepatitis than C34, respectively.

Claims

1. A pharmaceutical composition comprising ibudilast, dipyridamole, or both as active ingredients, for the treatment or prevention of Epstein-Barr virus hepatitis in a subject.

2. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises ibudilast.

3. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises dipyridamole.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the subject is a subject having a primary infection with the Epstein-Barr virus.

5. The pharmaceutical composition according to any one of claims 1 to 3, wherein the subject is a subject having infectious mononucleosis.

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