JPH03264530A - Anti-retrovirus agent - Google Patents

Abstract

PURPOSE:To obtain an anti-retrovirus agent effective in inhibiting reverse transcriptase activity to suppress the proliferation of virus by using a specific compound having porphyrin skeleton, phthalocyanine skeleton or protoporphyrin skeleton as an active component. CONSTITUTION:The objective agent contains a compound of formula I (R1 to R4 are sulfonyl or H), a compound of formula II (S1 to S4 are H, sulfonyl or group of formula III; S5 is bivalent metal ion), a compound of formula IV (T1 and T5 are ethyl or vinyl; T2 and T4 are carboxyethyl or carboxyl; T3 is H or carboxymethyl; T6 is bivalent metal ion) or their pharmacologically allowable salt as an active component. The compound is administered as an oral drug at a daily dose of 10mg to 7g in several divided doses or as a parenteral drug such as intravenous injection, drip infusion, subcutaneous injection and intramuscular injection at a daily dose of 0.5mg to 1.5g.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to an antiretroviral agent effective in treating various viral diseases caused by retroviruses.

[Prior Art and Problems] As research on viruses continues, treatments for viral diseases are gradually being established.

In particular, acquired immunodeficiency syndrome (AI) has become a problem recently.
HIV (Human Immunitis), which causes DS).

Deficiency Virus) is known as a retrovirus.

Retroviruses contain RNA-dependent DNA within their virus particles.
It is a virus that contains reverse transcriptase (hereinafter referred to as reverse transcriptase) and propagates as follows.

■After infecting host cells, viral RNA is first transcribed into DNA by reverse transcriptase.

(2) This DNA is integrated into the host cell chromosome, and then mRNA is synthesized by RNA synthesis in the host cell.

■Various virus proteins are produced by this mRNA.

There has been a desire to develop a drug that has an epoch-making therapeutic effect on human diseases caused by this retrovirus.

[Means for Solving the Problems] The present inventors conducted research on retrovirus growth inhibition effects on various compounds having a porphyrin skeleton, a phthalocyanine skeleton, and a protoporphyrin skeleton. The present inventors have discovered that several compounds having a porphyrin skeleton have retrovirus growth inhibiting effects, leading to the completion of the present invention.

That is, the present invention is as follows.

(1) An antiretroviral agent containing a compound represented by the following formula (R1, R2, R3 and R4 represent a sulfonyl group or a hydrogen atom) and a pharmacologically acceptable salt thereof as an active ingredient.

(2) An antiretroviral agent containing a compound represented by the following formula II and a pharmacologically acceptable salt thereof as an active ingredient.

(3) The following formula III (Sl, S2, S3 and S4 are hydrogen atoms, groups, or the following formula II-1 sulfonyl (T1 and T5 represent an ethyl group or a vinyl group, T
2 and T4 represent a carboxyethyl group or a carboxyl group, T3 represents a hydrogen atom or a carboxymethyl group, and T6 represents a divalent metal ion. ) An antiretroviral agent containing a compound represented by the following and its pharmacologically acceptable salt as an active ingredient.

S5 represents a group represented by: ) Compounds of divalent metal ions represented by formulas 1 to III are hereinafter referred to as compounds of the formulas.

Specific examples of compounds of the formula include the following compounds.

Compound A (αILγ, δ-tetraphenylporphine tetrasulfonic acid disulfonic acid salt tetrahydrate) Compound B (kappa phthalocyanine-3,4',4",4"2 tetrasulfonic acid tetrasodium salt) Compound C, phthalocyanine tetrasulfonic acid, tetrasodium salt) Compound E (chlorophyllin, covered, trisodium salt) Compound D (Reactive Blue 15) Compound F (hemin) -11= Above The compounds were manufactured by Kagaku Kenkyusho Co., Ltd. and Aldrich Chemical Company (Aldrich Chemical Campa).
It is commercially available from Ny Inc.).

The compound of the formula has been used for colorimetric determination of metals, staining, etc., but it was completely unknown that it had an antiretroviral effect.

Next, the anti-retroviral effect of the compound of the formula will be explained with reference to experimental examples.

Experimental Example 1 <Anti-AIDS virus activity test using 5UP-T1 cells (T cell 1ine system)> 5UP-TI cells (I x 104 cells) are placed per hole in a 96-well microplate. Next, add the serially diluted test substance to each formula. Then, add 10 to 20 TCID50 (50% of Ti5sue) of human immunodeficiency virus (H9/HTLV-III B cell supernatant) to each of these formulas.
Culture InhibitionDose) and infect it. This plate was heated at 3TC under 5% CO2 for 7 days.
After culturing for 12 days, the cytotoxicity was measured by the MTT method, and the maximum inhibition rate and inhibitory concentration of each test substance were calculated. The results are shown in Table 1.

Table 1 Experimental Example 2 <Anti-AIDS virus activity test using MT-4 cells (T cell 1ine system)> MT-4 cells (T
2×104 cells (cell 1ine system) were added. Next, add the serially diluted test substance to each formula. Each of these expressions is associated with human immunodeficiency virus (H9/HTLV-III).
B cell supernatant) was added to 10-20 TCID5Q (50% o
f Ti5sue Culture Inhibitio
n Dose) and infect it. This plate is 3T
After culturing for 5 days under C5% CO2, cytotoxicity was measured by the MTT method, and the 50% inhibitory concentration (IC5Q) for each test substance shown below was calculated. The results are shown in Table 2.

Table 2 Experimental example 3 Effect on reverse transcriptase activity> A reaction mixture having the following composition was prepared.

○Reverse transcriptase (Human immunodeficiency)
ency virus reverse transcr
iptase) 1 unit/ml ○ Polyadenylic acid/oligothymidylic acid complex as template/primer complex [polyadenylic acid pol
yrA, oligothymidylic acid pdT12-18 (1:
1)] Manufactured by Pharmacia, 5 parts 1 rnl Tris-hydrochloric acid (pH 8,0) 25 times. l○Potassium chloride '15mmol○Magnesium chloride 8mmol○Dithiothreitol 2mmol○[3H]deoxythymidine triphosphate (83,OCi/mmot) (1,0mC1/mt)
0.2pmot○deoxythymidine triphosphate
9.8□ol○ Test substance 0.03~
1100p/ml○Purified water Appropriate amount
P15- 16- (deoxynucleic acid triphosphate) is a specific activity unit that consumes 1□ol.

This reaction mixture 5011 was reacted at 37° C. for 30 minutes.

The reaction product DNA was quantified using the ion exchange filter method [3
The incorporation of radioactivity of deoxythymidine triphosphate into DNA was measured using a Beckman scintillation counter, and was determined as reverse transcriptase activity.
The 0% inhibitory concentration (Ic5o) was calculated. The result is the third
Shown in the table.

Table 3 Experimental Example 4 Inhibitory effect on transformation of primary chicken embryo fibroblasts by Crowus sarcoma virus (R8V)> Place 3.5 x 105 cells of primary chicken embryo fibroblasts per hole in a 24-well microplate, 6 at 37℃, 5% CO2
Cultured for hours. After culturing, R8V-8R-A strain, multiplicity of infection (M, O, 1,) approximately 2.0X10-4, and each test substance were added to the cells, and after culturing for 2 hours, the cells were washed, and each test substance was added to the cells. Add the test substance and agar medium, and store at 37°C, 5% CO
After culturing for 8 days under 2, the number of foci was measured,
The 50% inhibitory concentration (IC50) for each test substance was calculated. The results are shown in Table 4.

From the results of Experimental Examples 1.2.3 and 4 in Table 4, it was confirmed that the compound of the formula, which is the active ingredient of the antiretroviral agent of the present invention, has an antiretroviral effect based on the inhibitory effect on reverse transcriptase activity. Ta.

That is, the compound of the formula suppresses the proliferation of retroviruses by inhibiting the reverse transcriptase activity necessary for their proliferation, and therefore can be applied to any retrovirus.

Specific examples of retroviruses include leukemia virus, sarcoma virus, breast cancer virus, visna virus, maedi virus, and HIV (Human Immun. virus).

Deficiency Virus), HTLV-I (
Human T cell Leukemia Viru
s Type I), etc.

Next, the results of acute toxicity experiments will be shown to prove that the compound of the formula has low toxicity and high safety.

Acute toxicity experiments were conducted using test substances A, B, C, D and E1.
00 mg/kg was intraperitoneally administered to 5 mice each, and the number of surviving mice was examined after 7 days. As a result, no deaths were observed for test substances A, B, C, D, and E.

The results of the above acute toxicity experiments showed that these compounds have low toxicity and high safety.

Next, the dosage and formulation of compounds of formula will be described.

The compounds of the formula can be administered to animals and humans neat or with conventional pharmaceutical carriers.

The dosage form is not particularly limited and may be selected and used as required, including tablets, capsules, granules, fine granules,
Examples include oral preparations such as powders, parenteral preparations such as injections, and suppositories.

In order to achieve the desired effect as an oral agent, adults should usually take 10 mg to 7 g of the compound of the formula in divided doses several times a day, although this will vary depending on the age, weight, and severity of the disease of the patient. seems appropriate.

In the present invention, oral preparations such as tablets, capsules, and granules are manufactured according to conventional methods using, for example, starch, lactose, sucrose, mannitol, carboxymethyl cellulose, corns 19-20-turner, inorganic salts, and the like.

In addition to the excipients mentioned above, binders, disintegrants, surfactants, lubricants, fluidity promoters, flavoring agents, coloring agents, fragrances, and the like can be used in this type of preparation as appropriate. Specific examples of each are shown below.

[Combined starch, dextrin, gum arabic powder, gelatin,
Hydroxypropyl starch, methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant 1 Starch, hydroxypropyl starch, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, carboxymethyl cellulose, low substituted hydroxypropyl cellulose.

[Surfactant] Sodium lauryl sulfate, soy lecithin, sucrose fatty acid ester, polysorbate 80° [Lubricant 1: Talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate , polyethylene glycol.

[Fluidity promoter] Light anhydrous silicic acid, dry aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The compounds of the formula can also be administered as suspensions, emulsions, syrups, and elixirs, and these various dosage forms may contain flavorings and colorants.

In order to exert the desired effect as a parenteral agent, an adult dose of 0.5 mg to 1.5 g per day of the compound of the formula is usually required, although it varies depending on the age, weight, and severity of the disease of the patient. Injection, intravenous drip, subcutaneous injection, and intramuscular injection are considered appropriate.

This parenteral preparation is manufactured according to a conventional method, and generally uses distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc. as a diluent. be able to. Furthermore, a bactericide, a preservative, and a stabilizer may be added as necessary. In addition, from the viewpoint of stability, this parenteral preparation may be filled into a vial or the like, then frozen, water removed by ordinary freeze-drying techniques, and a liquid preparation prepared from the freeze-dried product immediately before use. Furthermore, isotonizing agents, stabilizers, preservatives, soothing agents, etc. may be added as appropriate.

Other parenteral preparations include liquid preparations for external use, liniments such as ointments, mold preparations for rectal administration, etc., and are manufactured according to conventional methods.

Next, a manufacturing example will be shown.

Example 1 ■Corn starch 44g ■Crystalline cellulose 40g ■Carboxymethyl cellulose calcium 5g ■Light anhydrous silicic acid 0.5g ■Magnesium stearate 0.5g ■Compound A1 total 100g Mix ■～■ uniformly according to the above recipe and press into tablets. Compression molding was performed using a machine to obtain tablets each weighing 200 mg.

This tablet-tablet contains 20 rng of compound A, and adults should take 3 to 10 tablets a day in several doses.

23-24 - Example 2 ■ Crystalline cellulose 84.5 g ■ Magnesium stearate 0.5 g ■ Carboxymethyl cellulose calcium 5 g ■ Compound B1 total 100 g According to the above recipe, ■, ■, and a part of ■ were uniformly mixed and compression molded. After that, the mixture was pulverized, the remaining amounts of (1) and (2) were added and mixed, and the mixture was compressed using a tablet machine to obtain tablets each weighing 200 mg.

This tablet-tablet contains compound B2orng, and adults should take 3 to 10 tablets a day in several doses.

Example 3 ■ Crystalline cellulose 34.5 g ■ 10% hydroxypropyl cellulose ethanol solution 50 g ■ Carboxymethylcellulose calcium 5 g ■ Magnesium stearate 0.5 g ■ Compound C1 total 100 g Mix ■ and ■ according to the above recipe uniformly, After the mixture was wetted by a conventional method, granulated by an extrusion granulator, dried and crushed, (1) and (2) were mixed and compressed by a tablet machine to obtain tablets each weighing 2 oomg.

These tablets contain the compound c2orng, and are taken by adults in 3 to 10 tablets a day in several doses.

Example 4 ■Corn starch 84g ■Magnesium stearate 0.5g ■Carboxymethylcellulose calcium 5g ■Light anhydrous silicic acid 0.5g ■Compound D1 Total 100g Mix ■~■ uniformly according to the above recipe and compress with a compression molding machine. After molding, the mixture was crushed using a crusher and sieved to obtain granules.

1 g of this granule contains 100 mg of Compound D, and adults should take 0.6 to 2 g in several doses per day.

Example 5 ■ Crystalline cellulose 55 g ■ 10% hydroxypropyl cellulose ethanol solution 35 g ■ Compound E1 Total 100 g According to the above recipe, ■ to ■ were mixed uniformly and made into a paste. After granulation using an extrusion granulator, the mixture was dried and sieved to obtain granules.

This granule 1. contains compound E100. It contains 0.6 to 2 g per day for adults, divided into several doses.

Example 6 ■Corn starch 89.5g ■Light silicic anhydride 0.5g ■Compound F1 Total 100g Mix ■~■ uniformly according to the above recipe and give 200mg
was filled into a No. 2 capsule.

27 28- One capsule of this preparation contains 20 mg of compound F, and adults should take 3 to 10 capsules a day in several doses.

Example 7 ■ Distilled water for injection Appropriate amount ■ Glucose 200 mg ■ Compound B
10mg total amount 15m
1. After dissolving ■ and ■ in distilled water for injection, they were poured into a 5 mt ampoule and autoclaved at 121°C for 15 minutes to obtain an injection.

Claims (3)

[Claims] (1) Compounds represented by the following formula I ▲ Numerical formulas, chemical formulas, tables, etc.▼ I (R_1, R_2, R_3 and R_4 represent a sulfonyl group or a hydrogen atom) and their pharmacologically acceptable compounds An antiretroviral agent whose active ingredient is salt. (2) Formula II below ▲There are mathematical formulas, chemical formulas, tables, etc.▼II (S_1, S_2, S_3 and S_4 are hydrogen atoms, sulfonyl groups, or the following formula II_-_1 ▲There are mathematical formulas, chemical formulas, tables, etc.▼II_ An antiretroviral agent containing a compound represented by -_1 and a pharmacologically acceptable salt thereof as an active ingredient.S_5 represents a divalent metal ion. (3) Formula III below ▲ Numerical formulas, chemical formulas, tables, etc. or carboxymethyl group, and T_6 represents a divalent metal ion.