WO2001036391A1

WO2001036391A1 - Thiamin disulfide derivatives

Info

Publication number: WO2001036391A1
Application number: PCT/JP2000/007431
Authority: WO
Inventors: Shozo Shoji; Kuniomi Tachibana
Original assignee: Nissui Pharmaceutical Co., Ltd.
Priority date: 1999-11-18
Filing date: 2000-10-24
Publication date: 2001-05-25
Also published as: AU7955900A; JP2001139557A

Abstract

Thiamin disulfide derivatives represented by general formula (1) or salts thereof and drugs containing the same as the active ingredient, wherein X?1 and X2¿ are the same or different and each represents hydrogen or halogeno; and R1 represents linear or branched alkylene. The compounds (1) or salts thereof are free from the problem of hardly water-soluble properties, which is a shortcoming of the conventional thiamin disulfides, and has a potent anti-HIV activity, thereby being useful as remedies for HIV-infection, in particular, remedies for AIDS.

Description

Description Thiamine disulfide derivative Technical field

The present invention relates to a thiamine disulfide derivative useful as a medicament for treating HIV infection. Background art

AIDS is a disease caused by HIV (Human Immunodeficiency Virus) infection, and the number of patients has been increasing rapidly since its discovery in the United States in 1983. Known therapeutic agents for AIDS include anti-HIV agents such as azidothymidine (AZT) and didanosine (DDI).

However, although AZT has a remarkable life-prolonging effect, it has the problem that side effects include headache, gastrointestinal disorders, and bone marrow suppression. In addition, many of these anti-HIV drugs, which have been studied in the past, are based on the mechanism of action that inhibits DNA synthesis during the replication process of HIV and suppresses the growth of HIV. At the same time, it also suppressed the DNA synthesis of normal cells, resulting in a decrease in the number of normal cells and an increasingly serious situation for patients. Disclosure of the invention

Accordingly, an object of the present invention is to provide a novel compound which is excellent in safety and anti-HIV action and is useful as an AIDS therapeutic agent.

Under such circumstances, the present inventors have screened the anti-HIV activity of various compounds using the recently developed screening method for anti-HIV drugs by suppressing Tat (transactivator). Widely used It has been found that thiamine disulfide and its 0-acylated product exert an anti-HIV effect by an action mechanism which is not present in existing anti-HIV drugs, and a patent application was previously filed (WO 94/02124, WO 98/20877). . However, these thiamine disulfides have a problem that they are poorly water-soluble. Thus, the present inventors have further studied and found that a compound obtained by converting two hydroxyethyl groups of thiamine disulfide into a hydroxyalkylaminoethyl group has excellent water solubility and a strong anti-HIV activity. And found that the present invention was completed.

That is, the present invention provides the following general formula (1)

(Wherein X ¹ and X ² are the same or different and each represent a hydrogen atom or a halogen atom, and R ¹ represents a linear or branched alkylene group)

A thiamine disulfide derivative represented by or a salt thereof, a medicament containing the compound as an active ingredient, a pharmaceutical composition containing the compound and a pharmaceutically acceptable carrier, use of the compound as a medicament, and It is intended to provide a method for treating HIV infection by administering the compound. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a diagram showing a mass spectrum of the compound (TDH) obtained in Example 1, FIG. 2 is a diagram showing a mass spectrum of the compound obtained in Example 2, and FIG. FIG. 1 shows anti-Tat activity. BEST MODE FOR CARRYING OUT THE INVENTION

In the general formula (1) representing the compound of the present invention, examples of the halogen atom represented by X ¹ and X ² include a chlorine atom, a bromine atom, a fluorine atom and an iodine atom, and a chlorine atom is particularly preferred. X ¹ and X ² are the same or different and are particularly preferably a hydrogen atom or a chlorine atom. As the alkylene group represented by R ¹ , a linear or branched one having 2 to 16 carbon atoms is preferable, and a linear alkylene group having 2 to 16 carbon atoms is more preferable. Specific examples of Ri include an ethylene group, a trimethylene group, a tetramethylene group, a hexamethylene group, an octamethylene group, a decamethylene group, a dodecamethylene group, and a tetradecamethylene group, and a hexamethylene group is particularly preferable. The salt of the compound (1) of the present invention is not particularly limited as long as it is a pharmaceutically acceptable salt, but acid addition salts, especially inorganic acid salts such as hydrochloric acid, nitric acid, and sulfuric acid, acetic acid, butyric acid, and succinic acid And the like are preferred. The compound (1) of the present invention or a salt thereof may exist as a solvate such as a hydrate, or may form a complex with other components. The compound (1) of the present invention or a salt thereof can be produced, for example, according to the following reaction formula.

(2)

(Four)

(In the formula, R ² represents an alkyl group, a fluoroalkyl group or a p-methylphenyl group, and X ¹ , X ² and R ¹ are the same as described above.)

That is, a thiamine disulfide (2) is reacted with a sulfonylating agent (3) such as trifluoromethanesulfonyl chloride to obtain a compound (4), and then a hydroxyalkylamine (5) is added thereto. By the reaction, a hydroxyalkylaminated thiamine disulfide derivative (1) is obtained. Examples of the sulfonylating agent (3) include methanesulfonyl chloride, trifluoromethanesulfonyl chloride, p-toluenesulfonyl chloride and the like. The reaction of the thiamine disulfide (2) with the sulfonylating agent (3) can be carried out in the presence of a base such as triethylamine in a solvent such as acetonitrile at a temperature of 0 to 60 ° C. Good. The reaction of the compound (4) with the hydroxyalkylamine (5) may be performed in a solvent such as, for example, acetonitrile at a temperature of 0 to 60 ° C.

In order to isolate the target substance from the reaction mixture, it is preferable to carry out washing, extraction, recrystallization, chromatography and the like in an appropriate combination.

The thus-obtained compound (1) or a salt thereof has high solubility in water, has excellent anti-HIV activity against persistently infected cells of HIV, and is useful for treating HIV infection and AIDS. It is useful as a drug such as an agent.

The medicament of the present invention may contain, as necessary, an excipient, a binder, a lubricant, a disintegrant, a coating agent, an emulsifier, a suspending agent, a solvent, a stabilizer, an absorbent, Pharmaceutically acceptable carriers such as adjuvants and ointment bases may be added as appropriate, or ribosome formation etc. may be formulated in dosage forms for oral administration, injection, rectal administration, etc. according to the usual methods. This is obtained.

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

The dose varies depending on the administration route, the age of the patient, the symptoms, and the like. Administer separately.

In addition, the medicament of the present invention can be used in combination with other anti-HIV agents represented by adithymidine and dysinosin, and these other anti-HIV agents can be incorporated into the medicament of the present invention. EXAMPLES Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.

Example 1

1 g of thiamine disulfide was placed in a 50 ml tube, and 8 ml of acetonitrile was added and mixed uniformly. To the mixed solution was added a total amount of 5 g of trifluoromethanesulfonyl chloride mixed well with 3 ml of acetonitrile, and after stirring, a mixed solution of 1.4 ml of triethylamine and 1.4 ml of acetonitrile was gradually added dropwise with stirring. The solution was stirred until the color of the solution became transparent (about 25 minutes), and when the solution became transparent, a solution of 1.25 g of acetonitrile in 6-aminohexanol prepared in advance was added dropwise with stirring. The reaction was carried out with shaking in a warm bath at 37 ° C for 1.5 hours. After the reaction, the same volume of acetonitrile-saturated n-hexane was added to remove unreacted and by-products, and the intermediate layer solution was removed. After leaving at 4 ° C for a while, the resulting precipitate layer was removed, and the supernatant was stored at 1-20. Remove the resulting white precipitate, add 2 volumes of water to the supernatant, collect the resulting mucous precipitate and supernatant, wash the mucous precipitate twice with an equal volume of water, and freeze-dry the supernatant to mucus After washing the aqueous solution similarly, it was purified by reversed-phase HP LC. The obtained solution was analyzed by MALD I-TOF-MS. As a result, a compound in which the hydroxy group of thiamine disulfide was substituted with a hydroxyhexylamino group (X ¹ = X ² = H, R ′ in the general formula (1)) =-(CH ₂ ) ₆ —; hereinafter abbreviated as “TDH”) (Fig. 1). This compound was water soluble.

Example 2

In the same manner as in Example 1 except that dichlorothiamine disulfide (X ^ X ^ Cl) in the general formula (2) was used instead of the thiamine disulfide in the example 1, Xi in the general formula (1) = X ² = C to give a compound of R '=-(CH ₂ ) _s —. Figure 2 shows the mass spectrum of this compound.

Test example 1

Measurement of HIV-1 anti-Tat activity (MAG I ZCCR 5—T atass ay) (principle)

Tat is a viral transcriptional activation protein that is produced at an early stage when proviral DNA is transcribed during HIV infection. Tat has the ability to bind RNA and binds to a sequence having a stem-loop structure immediately below the transcription start site, t a r (T a t a c t i v a t in g r e g i o n), and enhances translation. It also has the function of promoting elongation by acting on RNA polymerase. When HIV-1 recombinant Tat is administered from outside the cell to HeLa cells into which a gene having the HIV-1 LTR has been introduced upstream of the gene La-Z expressing β-ga1, Tat Due to the transcriptional activity of HIV-1, the enzyme —ga1 is expressed from LacZ, which is encoded downstream of HIV-1 LTR, and after 48 hours, the cells are immobilized and the added substrate X_ga1 is degraded. As a result, HIV_1 infected cells or cells transcriptionally activated by HIV-1 Tat are stained blue. The stained cells were counted under a microscope and used as an indicator of the transcriptional activity of HIV-lTat.

(Method)

HeLa cells transfected with the gene encoding HIV-1 shochu and acZ were fractionated into 1 × 10 ⁴ cellsZml in a 48-well plate and cultured for 24 hours. After removing the culture medium, the cells were cultured in the presence and absence of TDH (0-; L00M) for 37 and 60 minutes, and then added with HIV-1 recombinant Tat (6 Opmol wells) for 48 hours. Cultured in a well. After culturing, they were fixed and stained. Stained cells, which are indicative of HIV-1 infection, are determined under a microscope, and their relative activities to control experiments are determined.

(%), Each of which is shown as the average of two experiments.

(Result)

TDH is a water-soluble compound that overcomes the drawback of the thiamine derivative's poor solubility in water.When MAG IZCCR 5-Tassay was performed, as shown in Fig. 3, TDH was reduced to about 50 M at a concentration of 50 M. It showed 50% HIV-1 anti-Tat activity. Industrial applicability

The compound (1) of the present invention or a salt thereof overcomes the poor water solubility which is a drawback of conventional thiamine disulfides, and has a strong anti-HIV activity. Useful as an agent.

Claims

The scope of the claims

1. The following general formula (1)

Or a salt thereof.

2. The thiamine disulfide derivative according to claim 1, wherein R ¹ is a hexamethylene group, or a salt thereof.

3. A medicament comprising the thiamine disulfide derivative according to claim 1 or 2 or a salt thereof as an active ingredient.

4. The medicament according to claim 3, which is a therapeutic agent for HIV infection.

5. A pharmaceutical composition comprising the thiamine disulfide derivative according to claim 1 or 2, or a salt thereof, and a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, which is a therapeutic composition for HIV infection.

7. Use of the thiamine disulfide derivative or a salt thereof according to claim 1 or 2 as a medicament.

8. The use according to claim 7, wherein the medicament is a therapeutic agent for HIV infection.

9. A method for treating HIV infection, which comprises administering the thiamine disulfide derivative according to claim 1 or 2 or a salt thereof.