WO1996033712A1

WO1996033712A1 - Remedy for acquired immunodeficiency syndrome

Info

Publication number: WO1996033712A1
Application number: PCT/JP1996/001106
Authority: WO
Inventors: Masanori Baba; Ikuro Muruyama; Hiroyuki Masayasu
Original assignee: Daiichi Pharmaceutical Co., Ltd.
Priority date: 1995-04-25
Filing date: 1996-04-24
Publication date: 1996-10-31
Also published as: AU5513796A

Abstract

A remedy for AIDS and infection with HIV which comprises 2-phenyl-1,2-benzisoselenazol-3(2H)-one or its salt as the active ingedient. This drug is excellent in the effects of suppressing HIV-1 infection and suppressing the proliferation of HIV-1 in a cell line persistently infected with HIV-1 by a cytokine such as TNF-α and has a high safety, which makes it useful in the treatment of AIDS.

Description

Description Treatment for acquired immunodeficiency syndrome Technical field

The present invention relates to a therapeutic agent for acquired immunodeficiency syndrome (AIDS) and a therapeutic agent for HIV infection, which have excellent anti-HIV effects in both acute and persistent HIV infection systems. Background art

AIDS is a disease caused by HIV infection, the number of which has increased rapidly since its discovery in the United States in 1983 and is now widespread worldwide.

HIV, the virus that causes AIDS, includes type 1 (HIV-1) and type 2

(HIV-2). In particular, HIV-1 often has its gene mutated during viral replication, which makes it difficult to develop an effective vaccine, and inhibits reverse transcriptase, which is currently used as an anti-HIV-1 agent The emergence of drug-resistant viruses also hinders effective chemotherapy for drugs and protease inhibitors under development. Recent studies indicate that virus turnover in infected individuals

(t urn ove r) is extremely fast, and it has been reported that the virus in the body is replaced in about 2 weeks (Wei, X., Ghosh, S.K.,

Tay l or M. E. e t a 1.: Na t u r e, 373: 1 1 7-1 22

(1 995), Ho, D.D., Neumann, A.U., Per el s on, A.S.e ta 1 .: Natur e, 373: 123-126 (1 995)]. Such vigorous growth of HIV-1 suggests that even with effective antiviral agents, the virus mutates very quickly and acquires drug resistance. Azidothymidine (AZT) and didequininosine in reality

It has been reported that the use of (dd I) etc. diminishes efficacy in a few months and detects correspondingly resistant viruses to these drugs

[La rder, BA, Dar by, G. and Richman, D. D.. Sc i enc e, 243: 1731-1 734 (1 989),

L arder, B. A. and Kemp, S. D .: S cience, 246: 1 1 55-1 1 58 (1 989), St. C lair, MH, Martin, JL, Tudor-Wi Li ams, G. eta] .: Sc i enc e, 253: 1 557-1 559 (1 991)].

On the other hand, recent studies have increasingly revealed the fact that many host factors are involved in the formation and progression of the disease, AIDS. In particular, it has been reported that the expression and activation of the HIV-1 gene in infected cells are deeply related to various site forces and intracellular transcription factors (Fau ci, AS: Scienc). e, 262: 1101-1018 (1993)], in which TNF-hi remarkably enhances the proliferation of HIV-1 in T cells and macrophages, ie, TNF-hi transcriptional regulation. It has been shown to enhance the proliferation of HIV-1 through the factor NF-cB, and it is said that the state of intracellular redox such as active oxygen in the cell is deeply involved at this time. Duh, EJ, Maury, WJ, Folks, TM eta 1 .: Proc. Natl. Ac ad. Sci. USA, 86: 5974-5978 (19989), 0 sborn, L., Knkel, S. and Nabel, GJ: Proc. Natl. Ac ad. Sci. USA, 86: 2336-2340 (1 989)

Therefore, drugs that suppress the proliferation of HIV-1 may be new AIDS therapeutics. N-acetyl-L-cysteine (NAC) has been reported to have an inhibitory effect on the growth of HIV-1 [Ro ederer, M., Staa1, FJT, Raju, PAeta 1 .: Proc. Na USA, 87: 4884-4888 (1990)). Clinically tested as a therapeutic agent for AIDS.

However, NAC was weak in activity, required large-dose administration, and had a problem with increased side effects.

Accordingly, an object of the present invention is to provide a therapeutic agent for AIDS which has a high activity and effectively suppresses the proliferation of HIV-1 even at a low concentration. Disclosure of the invention

In view of such a situation, the present inventor conducted intensive research and found that Ebselen

(Ebse 1 en) 2-phenyl-1,2,1-benzisoselenazo-1-ru 3 (2H) 1-on-force Shows an excellent anti-HIV effect in acutely infected systems and is a persistently infected strain The present invention has been found to have an activity of suppressing TNF-mediated HIV proliferation and to be useful as a therapeutic agent for AIDS, thus completing the present invention.

That is, the present invention provides an AIDS therapeutic agent comprising 2-phenyl-1,2-benzisoselenazole-3 (2H) -one or a salt thereof as an active ingredient. The present invention also provides a therapeutic agent for HIV infection, comprising 2-phenyl-1,2-benbuisoselenazole-3 (2H) one (1) or a salt thereof as an active ingredient. Further, the present invention provides the use of 2-phenyl-1,2-benbuisoselenabul-13 (2H) -one or a salt thereof for producing an AIDS therapeutic agent. Further, the present invention provides the use of 2-phenyl-1,2-benzisoselenazoluru 3 (2H) -one or a salt thereof for the manufacture of a therapeutic agent for HIV infection. The present invention further provides a method for treating AIDS, which comprises administering to a patient an effective amount of 2-phenyl-1,2-benbuisoselenabul-3 (2H) one or a salt thereof. To provide.

Furthermore, the present invention provides a method for treating HIV infection, which comprises administering to a patient an effective amount of 2-phenyl-1,2-benzoisoselenabul-3 (2H) one or a salt thereof. It provides a method. BEST MODE FOR CARRYING OUT THE INVENTION

The 2-phenyl-1,2-benzoisoselenabul-13 (2H) one used in the present invention is a known compound, and has already been used as an antirheumatic agent (JP-A-57-56427), Agents (JP-A-57-67568), therapeutic agents for diseases caused by oxidative stress (JP-A-62-294613), therapeutic agents for liver diseases (JP-A-63-27431), malignant tumors Therapeutic agent (Japanese Patent Application Laid-Open No. 63-183528), renal disease therapeutic agent (Japanese Patent Application Laid-Open No. 64-56615), -1 311 13), Heart disease therapeutic agent (Japanese Patent Application Laid-Open No. 1-113114), Gastrointestinal disease therapeutic agent (Japanese Patent Application Laid-Open No. 1132522), radiation injury treatment (Japanese Patent Application Laid-Open (JP-A) No. 113577-18), an agent for infectious diseases (Japanese Patent Application Laid-Open No. 11-82025), and an agent for suppressing restenosis after percutaneous coronary angioplasty (Japanese Patent Application Laid-open No. No. 5,255,084). However, nothing is known about its effects on HIV.

The 2-phenyl-1,2-benzoisoselenazol-3 (2H) -one may be used as its salt or may be a hydrate.

2-phenyl-1,2-benzoisoselenazole-3 (2H) — (hereinafter referred to as “ebselen”) is preferred. This compound has the following formula in vivo:

(-S G indicates one S-gluthiothione group)

An active metabolite represented by Therefore, even if these compounds are administered, ebselen The same effect as described above can be obtained.

Ebselen or a salt thereof strongly suppresses cell death due to acute HIV infection, as shown in the test examples below. It also strongly suppresses HIV proliferation caused by TNF-stimulation in HIV persistently infected cells. Therefore, it is considered to be effective as a therapeutic agent for AIDS because it suppresses infection and onset of HIV.

In addition, the toxicity of Ebselen was extremely low as shown by the following LD ₅₀ (mg / kg) as a result of oral or intraperitoneal administration to mice and rats. In addition, there were no findings at the time of high-dose administration that would cause adverse effects.

table 1

The therapeutic agent for AIDS and the therapeutic agent for HIV infection of the present invention may be prepared by adding excipients, binders, lubricants, disintegrants, coating agents, emulsifiers, suspending agents, solvents, and solvents to ebselen or a salt thereof as necessary. Pharmaceutical carriers such as auxiliaries, absorption aids, and softening bases are added as appropriate, or riboforming is performed, and manufactured according to standard methods for oral administration, injection, rectal administration, etc. It can be obtained by making the agent.

Formulations for oral administration include granules, tablets, dragees, capsules, soft carbs, pills, solutions, emulsions, suspensions, etc. Formulations for injection administration include intravenous injection, muscle Suppositories, capsules, and the like are preferable as preparations for intrarectal administration, such as for intravenous injection, subcutaneous injection, and intravenous injection.

The dosage varies depending on the route of administration, age of the patient, symptoms, etc., but in general, for oral administration per adult daily, 100 to 2.0 mg of ebselen or a salt thereof, particularly 200 to 1 mg , 100 mg is preferable, and it is administered once or in several divided doses. Example

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto. Example 1 Anti-HIV-1 Effect on Acutely Infected Cells

Pau we 1 infected HIV-1 to MT-4 cells, a cultured cell line derived from CD4-positive lymphocytes, and measured the anti-HIV-1 effect of the drug based on the emerging cytopathic effect. MTT dye by the method of S. et al., CPauwe ls, R, Balza rini, J., Β aba, M. eta 1 .: J. Virol. Methods, 20: 309-321 (1988)] This was performed using

Specifically adjusts the MT- 4 cells 1 1 s / 1 X 1 0 5 ce, there to 2 of HIV one first Paiiota _beta strains, 000 CC ID ₅ o a (50% cellcultureinfecti ve do se) was added Infected. Next, this was poured into a micro-tie evening tray already containing ebselen diluted to various concentrations at a rate of 100 portions, and cultured at 37 ° C. As a control, cells not infected with the virus (uninfected cells: mock) were similarly cultured. On the fourth day, the cells were observed under a microscope, and the number of viable cells was measured by the MTT method.

The results are shown in Table 2 with the viable cell count of uninfected cells (moc k) in the absence of the drug as 100%.

Table 2 Drug concentration Number of living cells (%)

(M)

Infected cells (INFECTED) Uninfected cells (MOCK)

20 55. 8 66. 7

10 48. 3 90. 4

5 33. 1 1 09. 9

2.5 31. 7 1 12.0

1.25 26. 0 1 1 2.2

0 24. 0 1 00.0 As shown in Table 2, in the absence of the drug, HIV-1 infected cells were denatured and killed by more than 75% of uninfected cells due to the growth of the virus. The number of viable cells increased as the concentration of DNA was increased, and virus growth was suppressed. On the other hand, in uninfected cells, a slight decrease in the number of viable cells was observed at concentrations of 10 M or more. Force Observed under a microscope at 20〃M (Ebselen did not show any obvious cytotoxicity. Based on the above results, this drug has an inhibitory effect on the growth of HIV-1. I understand.

Example 2 Anti-HIV-1 Effect on Persistently Infected Cells

After infecting cells, HIV-1 is incorporated into the host DNA, and for some reason (site force, heat, ultraviolet light, etc.), transcription from the DNA occurs and multiplies. Whether ebselen has such an inhibitory effect on the proliferation of HIV-1 was determined by determining whether OM10.1 cells, which are persistently infected with HIV-1 cells and usually do not show virus proliferation, [Bu tera, ST, Perez, VL, Wu, B. — Y.eta 1 .: J. Virol., 65: 464 5-4653 (1991)].

Specifically, OM1 0. 1 cells 5 X 1 0 ⁴ ce 1 1

Then, after preincubation for 24 hours in the presence of various concentrations of ebselen, TNF—, a kind of cytokin, was added to a concentration of 10 UZm. After further culturing for 3 days, the amount of p24 antigen (proportional to the amount of virus) in the supernatant was measured using an ELISA kit. Table 3 shows the results.

Table 3

Anti-HIV-1 effect of ebselen on OM10.1 cells

As shown in Table 3, after culture for 3 days, the amount of p24 antigen in the supernatant was more than 2,000 times higher in the presence of TNF-free (PC) than in the absence (NC) of TNF-free. Increased. When Ebselen was used at a concentration of 20 mg, the amount of antigen in the supernatant was reduced to 43% of that in the presence of TNF alone (PC). On the other hand, under the microscope, even at 20 M, there was no apparent cytotoxicity of ebselen in OM10.1 cells. Based on the above results, this drug is dependent on TNF-H in persistently infected cell lines. It was found to have the effect of suppressing the growth of HIV-1. Industrial applicability

The therapeutic agent for AIDS and the therapeutic agent for HIV infection of the present invention are excellent in the inhibitory effect on HIV-1 infection and the inhibitory effect on HIV-1 proliferation by cytokines such as TNF-H in HIV-1 persistently infected cell lines, Because of its high safety, it is useful for treating AIDS.

Claims

The scope of the claims

1. A drug for the treatment of acquired immunodeficiency syndrome (AIDS) containing 2-phenyl-1,2-benzisoselenazoyl-3 (2H) -one or a salt thereof as an active ingredient.

2. A therapeutic agent for HIV infectious diseases comprising, as an active ingredient, 2-phenyl-1,2-benzoisoselenazole-3 (2H) -one or a salt thereof.

3. The therapeutic agent for infectious disease according to claim 2, wherein HIV is HIV-1.

4. Use of 2-phenyl-1,2-benzoisoselenabul-1 (2H) -one or a salt thereof for the manufacture of a therapeutic agent for acquired immunodeficiency syndrome.

5. Use of 2-phenyl-1,2-benzisoselenazole-3 (2H) -one or a salt thereof for the manufacture of a therapeutic agent for HIV infection.

6. The use according to claim 5, wherein the HIV is HIV-1.

7. A method for treating acquired immunodeficiency syndrome, which comprises administering to a patient an effective amount of 2-phenyl-1,2-benzoisoselenazoyl-3 (2H) -one or a salt thereof.

8. A method for treating HIV infection, which comprises administering to a patient an effective amount of 2-phenyl-1,2-benzoisoselenazoyl-3 (2H) -one or a salt thereof.

9. The method according to claim 8, wherein the HIV is HIV-1.