JPH0987193A - Anti-human immunodeficiency virus agent containing extract of ginger-family plant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an anti-human immunodeficiency virus agent effective for preventing the crisis of and healing acquired immunodeficiency syndrome, containing, as active ingredient, an extract of a ginger-family plant. SOLUTION: This agent is obtained by subjecting a plant of ginger family, such as Amomum aculeatum, Amomum foetens and Amomum hochreutineri of Amomum genus, and Nicolaia solaris and Nicolaia speciosal of Nicolais genus, to extraction with such a solvent as water, methanol, ethanol, benzene, toluene, pentane, or hexane. When water is to be used as extractant, it is added to the dried plant material at a rate of 1,000g of water per 50g of the dried plant material, and the extraction is continued at 25-100 deg.C for 0.5-24hr. This extract is made into a pharmaceutical preparation in the form of injection, extract, infusion, powder, capsule, supprository, or poultice, and administered at an oral dose of 100-1,000mg a day per adult.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an anti-human immunodeficiency virus agent, and more specifically to an anti-human immunodeficiency virus agent containing an extract of a ginger family plant.

[0002]

2. Description of the Related Art Human Immunodeficiency Virus
ficiency Virus: hereinafter referred to as HIV) is a type of retrovirus that causes abnormalities in the entire human immune system. When infected with HIV, the immune function is lowered, so that the disease is extremely susceptible to infection, and opportunistic infections such as carinii pneumonia and malignant tumors such as Kaposi's sarcoma are accompanied, and eventually death occurs. Therefore, this disease is called Acquired Immune Deficiency Syndrome (AIDS).

Currently, AIDS therapeutic agents used worldwide are AZT (3'-azido-2 ', 3'-dideoxythymidine), ddI (2', 3'-dideoxyinosine), ddC (2 ', 3'-
Dideoxycytidine). These are all nucleic acid compounds and reverse transcriptase inhibitors. However, it is necessary to administer AIDS therapeutic agents to patients for a long period of time, and there is a problem that these agents also inhibit normal DNA synthesis and cause serious side effects such as myelosuppressive action and carcinogenesis. In addition, there is a new problem of the emergence of resistant viruses, and combination therapy with drugs by another mechanism is expected for this.

[0004]

Since a therapeutic drug for AIDS is premised on long-term administration to patients, it is necessary to provide a drug which suppresses HIV proliferation and has few side effects. Further, administration of only a limited drug causes problems such as the emergence of drug resistant virus and enhancement of side effects due to increased dose, and it is necessary to develop various kinds of drugs.
An object of the present invention is to provide an anti-human immunodeficiency virus agent containing, as an active ingredient, an extract of a tropical ginger plant whose safety of raw materials has been confirmed.

[0005]

[Means for Solving the Problems] As a result of searching for plant-derived anti-HIV active substances, the present inventors have found that tropical ginger plants, particularly Amomum genus, Nicolaia genus (Nicolaia) We found that extracts of plants belonging to the genus Zingiber, the genus Hornstedtia, the genus Kaempferia or the genus Alpinia have the activity of suppressing HIV proliferation. And completed the present invention.

That is, the present invention is an anti-human immunodeficiency virus agent characterized by containing an extract of a ginger family plant. Further, in the present invention, the ginger family is a genus of Amomum, a genus of Nicolaia, a genus of Zingiber, and a genus of Hornstettia.
rnstedtia genus), Kaempferia genus (Kaempferia genus), or Alpinia genus (Alpinia genus).

In the present invention, the ginger family plant may be Amomum aculeatum, Amomum foetens, Amomum hochreutineri, Amomum maxima, Amomum maxima. Amomum
walang), Nicolaia solari
s), Nicolaia speciosa
a), Zingiber zerumbe
t), Hornstedtia molli
s), Kaempferia galanga
Alternatively, it is an anti-human immunodeficiency virus agent which is Alpinia sanderae.

A specific example of the tropical ginger family plant used in the present invention is Amom.
um aculeatum), Amomum foete
ns), Amomum hochreuti
neri), Amomum maxima, Amomum walang, Nicolaia solaris, Nicolaia speciosa, Zingiber Zernbeth (Z)
ingiber zerumbet), Hornstettia Morris (Ho
rnstedtia mollis), Kempferia galanga (Kaempf)
eria galanga), Alpinia saunderi (Alpinia sa)
nderae) and other plants. These plants are cultivated in the Southeast Asia-Malaysia region or grow naturally, and the rhizomes are used for food, spices, folk medicine and the like. The raw plant may be raw,
A dried product is preferred for storage and transportation, and a crushed product is preferred for extraction.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The extract of the present invention can be obtained as an extract by subjecting the rhizome or aerial part of a plant of the type described above to an ordinary extraction operation, that is, an operation such as extraction using a solvent. You can Alternatively, the extract thus obtained may be pulverized by a conventional means such as evaporation of the solvent under reduced pressure, freeze-drying, spray-drying, and alcohol precipitation.

In order to obtain an extract in the present invention, it is desirable to extract with water, but the extraction solvent is not limited to these, alcohols such as methanol and ethanol, benzene, toluene, chloroform, dichloroethane, pentane, hexane, Ethyl acetate, methyl acetate, ether and the like can be used. The extraction condition is not particularly limited, but when water is used, it is 100 to 10 per 50 g of dried plant material.
Add 00 ml and extract at a temperature of 25-100 ° C for 0.5-24 hours. The extraction may be repeated twice or more.

The anti-human immunodeficiency virus agent of the present invention can be formulated by a known technique and administered by parenteral administration such as intravenous injection or intramuscular injection, oral administration or transdermal absorption. The dosage form may be any suitable form for the administration method, and examples thereof include injections, extracts, infusions, powders, capsules, suppositories, and suppositories.

The dose of the anti-human immunodeficiency virus agent of the present invention varies depending on the symptoms, administration method, administration timing, administration period, age of patient, etc. About 1000 mg is suitable. However, the present invention is not limited by dose.

The anti-human immunodeficiency virus agent of the present invention comprises H
When an IV infection is suspected or confirmed, it can be administered to prevent the onset. Further, it can be administered as a therapeutic agent after the onset of AIDS.

[0014]

The present invention will be described in more detail with reference to the following examples. (1) Preparation of Extract Dried products of various plant materials shown in Table 1 and Table 2 were crushed with a grinder, and then sequentially extracted with chloroform, methanol and hot water. That is, 30 g of the above-mentioned dried and ground sample
Was first extracted with chloroform, and then the extraction residue was extracted with methanol. Finally, remove the methanol extraction residue by 2
Transfer to a liter beaker, add 300 ml of pure water and add 2
It was extracted by heating for an hour. After the heat extraction, suction filtration was performed to obtain a hot water extract and an extraction residue. A new solvent was added to the extraction residue and the same operation was repeated to perform hot water extraction three times in total. The hot water extracts thus obtained were combined and the solvent was distilled off under reduced pressure to obtain a hot water extract.

(2) HIV growth inhibitory activity MT-4 cells, a T cell line which is persistently infected with HTLV-1 (human T lymphocyte-tropic virus type I), has a high susceptibility to HIV and is susceptible to infection. After 3-5 days, a cytopathic effect (CPE) appears and most cells die.
CPE is a signature of viral growth and is easily observed by light microscopy. The HIV growth inhibitory effect of hot water extracts of various plants was examined by the following method.

HIV-1 (HTLV-I
IIB) at a ratio of 0.001 TCID ₅₀ / cell for 1 hour, and then RPM diluted with two-step diluted specimen
I1640 medium (containing 10% fetal bovine serum and antibiotics) was suspended at a concentration of 1.5 × 10 ⁴ cells / ml, and ² cells were used per well in a 96-well round bottom microplate.
Cultured in 00 μl. As a control, infected cells and non-infected cells suspended in a medium containing no drug were cultured.
On the 5th day of culture, cell growth and the degree of CPE inhibition were examined by microscopy. The HIV growth inhibitory effect was represented by the minimum effective concentration (EC) for inhibiting CPE, and the cytotoxicity was evaluated by the viability of cells, and represented by the minimum concentration (CC) showing cytotoxicity. Table 1 shows the results.

[0017]

[Table 1]

(3) Inhibition of multinucleated giant cell formation Molt-4 / HIV-1 persistently infected with HIV-1
When cells and non-infected Molt-4 cells are mixed and cultured, it is easily observed by an optical microscope that both cell membranes fuse with each other to form multinucleated giant cells. The following method was used to examine the effect of inhibiting the formation of multinucleated giant cells in a sample in which the HIV growth inhibitory effect was observed. In a flat-bottom 96-well microplate, 100 μl / we of a sample solution diluted to a predetermined concentration with a medium (containing 10% fetal calf serum and antibiotics)
ll added. Next, persistent infection Molt suspended in the medium
-4 / HIV-1 cell suspension (1 × 10 ⁶ cells /
ml) was added at 10 μl / well, and then non-infected Mol
t-4 cell suspension (1 × 10 ⁶ cells / ml) was added to 9
0 μl / well was added and stirred, and mixed culture was performed at 37 ° C. in the presence of 5% CO ₂ . 24 hours after the mixed culture, the presence or absence of multinucleated giant cells was examined under a microscope. The effect of inhibiting the formation of multinucleated giant cells was expressed as the minimum effective concentration (EC) that inhibits the formation of multinucleated giant cells. The results are shown in Table 2.

[0019]

[Table 2]

(4) Results From the results in Table 1, it is clear that Amomum acureatum (Amom)
um aculeatum), Amomum foetens, Amomum hochreutiner
i), Amom maxim
a), Amommwaran
g), Nicolaia so
laris), Nikolaia Speciosa (Nicol)
aia speciosa), Zingiber zerumbet, Hornstedtia molis (Hornstedia mol)
lis), Kaempfer Garanga
ia galanga), Alpinia Sanderei (A
HI to the hot water extract of Lpinia sanderae)
A V growth inhibitory action was observed. Further, none of these showed cytotoxicity at a concentration of 500 μg / ml. Next, from the results shown in Table 2, all of these specimens exhibited a multinucleated giant cell formation inhibitory action and were correlated with the HIV growth inhibitory activity. From this, it was revealed that the anti-human immunodeficiency virus agent of the present invention has an action of inhibiting the binding between HIV and cells, and therefore is effective as a preventive or therapeutic agent for the onset of acquired immunodeficiency syndrome.

(5) Acute toxicity test When an oral acute toxicity test was conducted on ICR mice, the acute toxicity (LD ₅₀ ) of the anti-human immunodeficiency virus agent of the present invention was 1000 mg / kg or more.

[0022]

INDUSTRIAL APPLICABILITY The tropical ginger family of the present invention is of the genus Amomum and genus Nicola.
ia), Zingiber genus (Zingiber genus), Hornstedtia genus (Hornstedtia genus), Kaempferia genus (Kaempferia genus) or Alpinia genus (Alpinia genus), the safety of the raw material has been confirmed. Therefore, it can be effectively used for a long time as an anti-human immunodeficiency virus agent that suppresses the proliferation of HIV.

Claims (3)

[Claims] 1. An anti-human immunodeficiency virus agent comprising an extract of a ginger family plant. 2. The ginger family is of the genus Amomum.
Genus), Nikolaia (Nicolaia), Zingiber (Zi
genus ngiber), Hornstedtia
Genus), Kaempferia genus or Alpinia genus, and the anti-human immunodeficiency virus agent according to claim 1. 3. The ginger family plants are Amomum aculeatum and Amomum photens.
omum foetens), Amomum
hochreutineri), Amomum maxim
a), Amomum walang, Nicolaia solaris, Nicolaia speciosa, Zingiber zerumbet, Hornstettia
The anti-human immunodeficiency virus agent according to claim 1 or 2, which is Morris (Hornstedtia mollis), Kaempferia galanga or Alpinia sanderae.