WO1993002671A1

WO1993002671A1 - Retrovirus proliferation inhibitor

Info

Publication number: WO1993002671A1
Application number: PCT/JP1992/000945
Authority: WO
Inventors: Takashi Tsuruo; Keisuke Yusa; Mitsugi Kozawa; Kimiye Baba; Hiroshi Hara
Original assignee: Taisho Pharmaceutical Co., Ltd.
Priority date: 1991-07-30
Filing date: 1992-07-24
Publication date: 1993-02-18
Also published as: AU2346292A

Abstract

A retrovirus proliferation inhibitor containing xanthoangelol represented by formula (I), as an active ingredient, which has such a low molecular weight that it is readily absorbable, is lowly toxic and inhibits the proliferation of retrovirus intensely, so that it can be used in preventing or treating diseases caused by retrovirus, such as AIDS, adult T-cell leukemia and HAM. Conceivable mechanisms of action of xanthoangelol include those other than the inhibition of reverse transcriptase, for example, the inhibition of syncytium formation (that of giant cell formation).

Description

Small

Specification

Retrovirus growth inhibitor Technical field

The present invention relates to a retrovirus growth inhibitor which is effective in preventing and treating diseases caused by a retrovirus such as an AIDS virus, and more particularly to a retrovirus growth inhibitor comprising xanthoangelol as an active ingredient. Related to inhibitors. Background art

The disease retro virus occurs because of AIDS, adult T-cell leukemia, HAM (HTLV one 1 A ssociated My elopathy) force there ^s. Therefore, if the growth of retrovirus can be inhibited, the above diseases can be prevented or treated. Among these diseases, AIDS is a disease that occurs in the case of HIV (Human Immu nodeficiency Virus), a type of retrovirus, and the method for evaluating its effects has been established by Yamamoto et al. [Sience 22 9 56 3 (1 1985)].

As an anti-AIDS drug, 3-azidode beef cytidine (ddI) has already been marketed. Compounds having an inhibitory effect on HIV reverse transcriptase, such as ddI, have serious side effects such as suppression of bone marrow formation, and such improvements are desired.

Also, dextran sulfate is known as having an inhibitory action on the syncytium formation of HIV (giant cell formation inhibitory action). However, this dextran sulfate has the disadvantage that when administered orally, it is degraded in the stomach and has poor absorption due to its high molecular weight.

The active ingredient of the present invention, bovine santoangelol, is a known compound and can be obtained from Ashitapa (Angericakeiskei K0IDZUMI) by Ozawa et al. In Pharmaceutical Magazine Vol. 98, No. 210 (19778). _c However, it has been reported on), it has not been reported for retro viral growth inhibitory effects of cattle San Tangeroru. Disclosure of the invention

The present inventors have sought to search for compounds having a retrovirus growth inhibitory effect on plant components widely, and as a result of diligently examining their activities, bovine santoangelol strongly inhibits the growth of retrovirus. Thus, the present invention has been completed.

Hereinafter, the present invention will be described in detail.

Xanthoangelol, which is an active ingredient of the present invention, can be widely used in the Aceritaceae family according to the method of

It is a compound represented by these.

The active ingredient of the present invention, xanthoangelol, showed almost no reverse transcriptase inhibitory activity. For example, the inhibitory activity of HIV on the reverse transcriptase is 205 / i% / m.1 in terms of ED 5B value, and the inhibitory activity of AMV (Avian My clobastosis Virus) on the reverse transcriptase is ED ₅₀ The value was 1000 gZm 1 or more. The mechanism of action of xanthoangelol may be a mechanism other than reverse transcriptase inhibitory action, for example, syncytium formation inhibitory action (giant cell formation inhibitory action).

The amount of xanthoangelol to be incorporated into the retrovirus growth inhibitor of the present invention is preferably 50 mgZ days to 100 mg days, more preferably 100 mg / B to 600 mg days. preferable.

As the administration form of the retrovirus growth inhibitor according to the present invention, various forms can be selected according to the purpose of treatment. Examples include oral preparations such as powders, syrups, tablets and granules, and parenteral preparations such as injections and suppositories. In shaping into these dosage forms, carriers conventionally known in this field can be used, and they are produced by means commonly used in this field. For example, commonly used excipients for oral preparations (lactose, glucose, mannitol, crystalline cellulose, etc.), disintegrants (potato starch, corn starch, low-substituted hydroxypropylcellulose, Carboxymethylcellulose, etc.), lubricants (light gay anhydride, talc, magnesium stearate, etc.) and binders (gelatin, arabian gum, polyvinyl alcohol, hydroxypropylcellulose, etc.). Yes, flavoring agents, flavors, preservatives, etc. can be added as needed. BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples.

Example 1

Tablets were produced by a conventional method according to the following formulation.

Xan Toange's mouth 0 0 mg

Lactose 60 0 mg

Cornstarch 17 mg

Microcrystalline cellulose 45 mg

Light gay anhydride 15 mg

Hydroxypropyl cellulose 10 mg

Magnesium stearate 3 mg

250 mg per tablet

Example 2

Capsules were produced according to the following formulation by a conventional method.

Xanthoangelol 200 mg

Lactose 3 5 mg

Low substituted hydroxypropylcellulose 40 mg

Magnesium aluminate metasilicate 5 mg

Hydroxypropyl pill cellulose 15 mg

Gan-dani castor oil 5 mg

Talc 10 mg

3 10 mg per tablet Test example 1 (anti-HIV activity measurement method)

(Preparation method of retrovirus growth inhibitor)

Xanthoangelol was dissolved in DMS0 to 200 mg / ml. This solution was serially diluted with DMS0 every 10 fold to prepare 7-step samples up to 0.2 g / 1.

(Culture of HIV cells)

Incubate HI Vl (HT L V-ΠΒ) infected MOL T-4 cells (1 × 10 ⁵ ce 11 s / m 1) for 3 days, remove the cells by centrifugation, Filtered, one-70. Virus solution that was stored at ^{C (1 X 1 0 5 P} FU / m 1, PFU: plaque forming unit) was used as a. RPMI 1164 medium containing 10% fetal bovine serum was used for cell culture.

(Test method)

M.0 the HI V-1 in MT-4 cells. I. (Multiplicity of infection) 0.0 0 2 after infection ^{with, 1.6 xi 0 5 cel 1 s} / m 1 and cells were U-bottom 9 6 Uweru plate 50/5 1 each.

1 n1 of the sample prepared in advance was added to 1 ml of the culture medium and stirred to obtain a test solution. Next, this test solution was added at a rate of 501 per cell, and cultured under the conditions of 5% C02 and 37 ° C, and the number of viable cells was counted after XTT (2,3-bis [2- methoxy-4-ni tro-5-sulf ophenyl] -5- [(p phenylaraino) -carbonyl] -2H-tetrazolium nydroxide). X TT solution [0.5 mgZm i XTT, 0.3 ^ g / 1 PMS (N-methylphenazo niira) ] each © - le 50 1 In addition 5% C0 _2, 37. After incubating for 4 hours under C conditions, the absorbance at 450 nm was measured.

(Judgment method)

Xanthoangelol anti-HIV activity, bovine San Toangeroru is HIV - it was assessed by 4 concentration inhibiting 50% cytopathic effect on the cells (ED ₅ B) - 1 of MT. That is, in the absence of xanthoangelol, the absorbance of virus-uninfected cells is set at 100%, and the absorbance of virus-infected cells is set at 0%. The concentration of xanthoangelol was determined. Test Example 2 (Effect on cells)

Xanthoangelol cytotoxicity was examined simultaneously with anti-HIV activity. HIV - 1 uninfected MT-4 cells ^{(1. 6 xl 0 5 cells /} 1) was dispensed at 5 0 1 minute U bottom 9 6 Uyurupure Bok. It was added followed by similarly each Ueru per 5 0 / l to test solution prepared in Test Example _{1, 5% C 0 2,} 3 7 ° cultured C conditions, measuring the number of viable cells by XTT method after 6 days did. The XTT solution was added to each table in 501, and incubated for 4 hours under the conditions of 5% CO ₂ and 37 ° C., and then the absorbance at 450 nm was measured. The absorbance of non-virus-infected cells in the absence of xanthoangelol was set at 100%, and the concentration of xanthoangelol (ID50) that inhibited this color development by ₅₀ % was determined. did.

The results of Test Examples 1 and 2 are summarized in Table 1 and FIG.

Test Example 3 (Acute toxicity test)

S prague--Dawley rats S (body weight: 190-230 g) were used as a group, and the results of observation of the course of 14 days after oral administration of xanthoangelol SOO mgZkg once orally No toxicity was observed. No abnormalities were observed during the subsequent autopsy, 14 said. The same dose was similarly observed after intraperitoneal administration and subcutaneous administration.

Five ICR mice S (body weight: 22.5 to 23.5 g) were used as a group, and 100 mg / kg of xanthrangerol was orally administered once daily for 3 days. As a result of observation, no toxicity was observed. No abnormalities were observed in the autopsy following the statement. The same dose was similarly observed intraperitoneally and subcutaneously, followed by observation over the same period. No toxicity was observed. b—brief description of the drawing

FIG. 1 shows the antiviral effect of xanthoangelol on virus-infected and uninfected cells at various concentrations. Industrial applicability

Xanthoangel, which is an active ingredient of the retroviral growth inhibitor of the present invention, is a low molecular compound, has no problem in its absorbability, has little toxicity, and strongly inhibits the growth of retrovirus. AIDS caused by the virus, adults

Diseases such as T cell leukemia and HAM can be prevented or treated.

Claims

The scope of the claims

Expression,

A retrovirus growth inhibitor containing xanthoangelol as an active ingredient c