KR100532542B1 - Pharmaceutical composition comprising arsenic acid ,meta-arsenite,and pharmaceutically allowable salts - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating AIDS containing arsenic acid, metaarsenite, or a pharmaceutically acceptable salt thereof, and more particularly, arsenic acid, which is an in vivo metabolite of arsenic trioxide. ), Meta-arsenite, and pharmaceutically acceptable salts thereof, exhibit activity against anti-HIV-1 and anti-HIV-2, and thus new pharmaceutical compositions for preventing and treating AIDS.

Description

Pharmaceutical composition for preventing and treating AIDS containing arsenic acid, metaarsenite or pharmaceutically acceptable salts thereof

The present invention relates to a pharmaceutical composition for preventing and treating AIDS containing arsenic acid, metaarsenite, or a pharmaceutically acceptable salt thereof, and more particularly, arsenic acid, which is an in vivo metabolite of arsenic trioxide. ), Meta-arsenite, and pharmaceutically acceptable salts thereof, exhibit activity against anti-HIV-1 and anti-HIV-2, and thus new pharmaceutical compositions for preventing and treating AIDS.

Since AIDS was first reported in San Francisco, USA in 1981, the number of newly infected patients has been around 5 million in 2002, and about 3 million people have died of AIDS.

It is estimated that the number of people infected with HIV reported to the World Health Organization by the end of December 2002 is 42 million.

In Korea, the number of people living with HIV has increased steadily since it was first confirmed in December 1985. As of the end of September 2003, the number of people living with HIV was 2,405.

With the rapid increase in HIV and AIDS patients, the United States, Canada, France, Japan, and other countries around the world have accelerated the development of AIDS treatments.At previously known anti-HIV substances, AZT (3-azido) has been approved by the US FDA. There are only a few such as -3-deoxythymidine, dideoxyinosine (ddI), and dideoxycytidine (ddC). Most of these compounds are currently used only in cells that are already infected. Because it does not inhibit viral replication, it only has the effect of prolonging life rather than treating the disease completely. In addition, it causes side effects such as a decrease in platelet count and a decrease in bone marrow blood cells. The problem is pointed out, such as a lot of viruses are found, the development of new drugs is required.

Therefore, while the inventors of the present invention have been searching for a new drug to solve the above problems, the in vivo metabolites of arsenic trioxide, arsenic acid, metaarsenite, and their pharmaceutically acceptable salts are anti-HIV-1 and anti-HIV-. The present invention was completed by confirming that the activity of 2 was shown.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing and treating AIDS, which contains arsenic acid, metaarsenite, or a pharmaceutically acceptable salt thereof.

The present invention is characterized by a pharmaceutical composition for preventing and treating AIDS, which contains one or two or more selected from arsenic acid, metaarsenite and pharmaceutically acceptable salts thereof as an active ingredient.

Referring to the present invention in more detail as follows.

The present invention shows that arsenic trioxide, an in vivo metabolite, arsenic acid, meta-arsenite, and pharmaceutically acceptable salts thereof, exhibit activity against anti-HIV-1 and anti-HIV-2. It relates to a pharmaceutical composition for preventing and treating AIDS.

Arsenic acid, which is an active ingredient of the pharmaceutical composition according to the present invention, is one of the in vivo metabolites of arsenic trioxide, and may be obtained from blood or organs in vivo or prepared by a synthetic method, and has been shown to have anticancer efficacy in leukemia and solid cancer. It is known.

In addition, metaalsenite, another active ingredient, is an in vivo metabolite of arsenic trioxide, such as arsenic acid, which may be obtained from blood or organs in vivo or manufactured by a synthetic method. To date, anti-cancer efficacy in leukemia, solid cancer, etc. It is known.

On the other hand, the arsenic acid and meta arsenite may be pharmaceutically acceptable salts of the arsenic acid and meta arsenite together with acids such as inorganic acids such as hydrochloric acid, acetic acid and bromic acid. And a pharmaceutically acceptable salt of arsenic acid and a pharmaceutically acceptable salt of metaarsenite, because both may react with alkali metal ions such as sodium and potassium to form pharmaceutically acceptable salts. Included in

The degree of inhibition of CPE (Cytopathic Effect) on MT-4 cells infected with HIV-1 and HIV-2 with the arsonic acid, metaarsenite and their pharmaceutically acceptable salts of the present invention was determined by MTT ( Arsenic acid, metaarsenite, and their pharmaceutically acceptable salts showed better antiviral activity and less cytotoxicity at the same concentration than AZT, which is currently used for the treatment of AIDS. Confirmed.

In addition, as arsonic acid, metaarsenite, and pharmaceutically acceptable salts thereof exhibit activity against anti-HIV-1 and anti-HIV-2, respectively, mixtures thereof are also excellent anti-HIV-1 and anti-HIV-2. It is obvious to show activity against.

Therefore, the present invention can be used as an active ingredient of the anti- AI treatment and preventive treatment of arsenic acid, metaarsenite and pharmaceutically acceptable salts thereof, in addition to the active ingredient in the manufacture of a pharmaceutical containing the same Possible carriers or excipients may be used in formulated form as a unit or multiple dosage form for tablets, powders, granules, capsules, suspensions, emulsions or parenteral administration.

The effective dose of the active ingredient represented by the composition may be varied depending on the age, physical condition, weight, etc. of the patient, preferably 0.1 to 30 mg / kg / day, more preferably 0.5 to 3 mg / It is administered within a kg / day range. In addition, within a daily effective dosage range is divided into once a day or several times a day.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Reference Example

(1) cell and virus lines

Human T cell transformed cell line MT-4 was used in culture in RPMI1640 medium containing 10% fetal bovine serum.

In addition, HIV-1 IIIB and HIV-2 strains were distributed from the National Institute for Biological Standards and Control (NIBSC) in the United Kingdom. Each virus was passaged at intervals of 3 to 4 days after infection with H9 cells, and centrifuged supernatants from which cells were removed were taken as virus seed stock, stored at -70 ° C, and used at 37 ° C immediately before the experiment.

(2) sample

Lepatox was provided with sodium arshenic acid salt and metaarsenite sodium salt, which were stored at 4 ° C. or lower. For in vitro studies, sodium arsenic and metaarsenite sodium salts were dissolved in DMSO and diluted in culture medium to a concentration of DMSO of 0.3% or less. Meanwhile, AZT was used as a standard drug.

(3) virus titer measurement

The viral titer of the virus solution used for the test was expressed as ₅₀ % tissue culture infection (CCID ₅₀ ) by titration by limiting dilution on MT-4 cells.

Example 1: in vitro Anti-HIV effects

In vitro experiments were carried out to investigate the anti-HIV effects of sodium arsenic salt and metaarsenite sodium salt.

1.0 × 10 ⁴ MTs infected with HIV-1 and HIV-2 to 100 μl of arsonic acid and metaarsenite sodium salts diluted twice-well per well into 96 well plates and inoculated to 100 CCID ₅₀ 100 μl of -4 cells were added and incubated in a CO ₂ incubator at 37 ° C. for 5 days, and then the number of surviving cells was compared with the CC (cell count) using MTT search and microscopic observation. The antiviral effect and cytotoxicity of metaarsenite sodium salt were measured. _{A; (IC 50 50% effective concentration} ) , the 50% of the non-infected with a virus (mocked infected) cells compared with the CC to the concentration of drug showing the antiviral effect of 50% of the cells to survive virus-infected EC ₅₀ The concentration of drug that was toxic to death was calculated as CC ₅₀ (50% cytotoxic concentration).

Comparison of Cytotoxic Effects of Each Sample sample CC ₅₀ (μg / ml) AZT 2.9 Arsenic acid sodium salt 6.0 Metaarsenite sodium salt 4.6

Comparison of antiviral (anti-HIV) effects of each sample sample IC ₅₀ (μg / ml) HIV-1 HIV-2 AZT 0.009 0.027 Arsenic acid sodium salt 0.007 0.0009 Metaarsenite sodium salt 0.003 0.0004

As shown in Table 1 and Table 2, the sodium arsenic acid salt and the metaarsenite sodium salt according to the present invention had superior antiviral activity at the same concentration than AZT, which is currently used for AIDS treatment, and had less cytotoxicity. It was confirmed.

Example 2: Subacute Toxicity Test in Mice

Subacute toxicity test by oral administration was performed using SPF CD-1 mice at 5 weeks of age.

The sodium arshenic acid salt and metaarsenite sodium salt of the present invention were dissolved in 10% glucose solution, respectively, and orally once daily for 14 days at a dose of 10, 20, 40, and 80 mg / kg / day for 10 animals in each group. Administered. After the recovery period for 14 days, biochemical and hematological examinations were performed on the 29th day, and autopsy was performed to observe the abnormalities of organs. In addition, the clinical symptoms, weight change, and death of animals were observed every day after administration of the test substance.

As a result, in the group administered 10, 20 and 40 mg / kg of the two test substances, there were no animals showing significant clinical symptoms or dead animals in all animals. Weight change, blood test, blood biochemical test, autopsy No change in toxicity was observed.

On the other hand, in the group of 80 mg / kg of sodium arsenic acid salt, one animal died on the 13th day after oral administration, and two animals developed abnormal symptoms such as dyspnea, but no more deaths occurred.

In addition, in the group administered 80 mg / kg of metaarsenite sodium salt, three died on the fourth day after oral administration, one died on the seventh day, and four of the others had abnormalities such as dyspnea. Symptoms occurred but did not die.

As a result of the subacute test of the present invention, when the oral sodium arsenic salt and metaarsenite sodium salt were orally administered to CD-1 mice, the toxicity was not changed up to 40 mg / kg, and the oral harmless dose was 40 mg / kg. It was determined to be a safe substance that is more than kg

Formulation Example 1 Preparation of Tablet

One tablet (200 mg) was mixed with 100 mg of active ingredient (arsenic acid, metaarsenite or a pharmaceutically acceptable salt thereof), 40 mg of lactose, 51 mg of corn starch, and 2 mg of colloidal silicon dioxide. 3% polyvinylpyrrolidone solution was added to the mixture and ground, and then passed through a 14 mesh sieve. After drying and passing the 14 mesh sieve again, the mixture obtained by adding 1 mg of magnesium stearate was made into a tablet.

Formulation Example 2: Preparation of Injection

In 10 ml of an ampoule, 10 mg of the active ingredient (arsenic acid, metaarsenite or a pharmaceutically acceptable salt thereof) is dissolved in 200 mg of polyoxyethylene hydrogenated castor oil, and distilled water for injection is made into 10 ml. Injections containing a concentration of 1 mg in 1 ml were prepared.

As described above, the pharmaceutical composition according to the present invention is effective in treating arsenic acid, metaarsenite or their pharmaceutically acceptable salts with low cytotoxicity and excellent activity against anti-HIV-1 and anti-HIV-2. It is expected to be very useful for the prevention and treatment of AIDS by using as an ingredient.

Claims (1)

A pharmaceutical composition for preventing and treating AIDS, characterized in that it contains one or two or more selected from arsenic acid, metaarsenite, and pharmaceutically acceptable salts thereof as an active ingredient.