KR100567456B1 - Extract of Kalopanax pictus having inhibitory effect on replication of human immunodificiency virus and therapeutic agent for AIDS containing the same - Google Patents

Abstract

The present invention relates to an extract of Yin-tree having HIV inhibitory activity and an AIDS therapeutic containing the same as an active ingredient. Yin-tree extract of the present invention is excellent in inhibiting HIV reverse transcriptase activity, inhibiting protease activity, inhibiting glucosidase activity and inhibiting HIV growth, and thus can be useful for treating AIDS, inhibiting progression, and inhibiting infection.

Description

Extract of Kalopanax pictus having inhibitory effect on replication of human immunodificiency virus and therapeutic agent for AIDS containing the same}             

1 is a result of HPLC analysis of a substrate confirming that the extract of the present invention inhibits the activity of HIV-I protease.

Figure 2 is a graph showing the HIV replication inhibitory activity of the lacquer extract of the present invention.

The present invention relates to an AIDS therapeutic agent, and more particularly, to an extract of Yin-tree having HIV growth inhibitory activity and an AIDS therapeutic agent containing the same as an active ingredient.

Human immunodeficiency virus type I (HIV-I), a species of retrovirus and an aetiological agent of AIDS, is genetically based on RNA (Kaminchik, J. et al ., AIDS res. hum.retroviruses, 1994, 10: 1003-1010; Peliska, JA et. al ., Science, 1992, 258: 1112-1118). HIV-I is a surface protein (glycoprotein 120, gp120) that interacts with the CD4 + receptor on helper T cells (Weiss, S. et. Al ., Gene, 1992, 111: 183-197; Hoxie, JA et. Al ., J. Immunol., 1986, 136: 361-363) Insert RNA into host cells and reverse transcriptase (RT) (Prasad, VR and Goff, SP Proc. Natl. Acad. Sci. USA, 1989, 86: 3104-3108), which are transcribed into viral DNA and present in the form of proviral DNA on the host chromosome. Viral mRNA is produced by RNA polymerase when the gene is expressed through the incubation period in the host cell, and processing of the viral protein is performed by protease (PR) (Katz, RA and Skalka, AM). , Annu. Rev. Biochem ., 1994, 63: 133-173). Viral surface glycoproteins are formed by the action of glucosidase (GL) (Taylor, DL, Antimicrobacterial Agents & Chemotherapy, 1994, 38 (8): 1780-1787; Mohan, P., Pharmaceutical Research, 1992, 9 (6): 703-714) Germinate into mature virus particles. As such, viruses are known to cause serious impairment of human immune system by inserting their genes into helper T cells and destroying host cells in the process of replicating themselves through incubation period.

As a chemotherapy to treat HIV-I, drugs such as sumarin are used (Mitsuya, H. et. Al ., Science, 1984, 226: 172-174), and reverse transcription of HIV-I Drugs for inhibiting enzymes include nucleoside analogs azidothymidine (AZT) (Ostertag, W. et. Al ., Proc. Natl. Acad. Sci. USA., 197, 471: 4980-4985), didanosine (ddI) and zalcitabine (ddC) are used, and non-nucleoside analogues TIBO, nevirapine and pyridinone are used in the clinic. have. However, AZT is myopathy (Schroder, JM et. Al ., Acta Neuropathologica, 1996, 92 (2): 138-149) and oral mucosal pigmentation (Tadini, G. et . , Archives of Dermatology, 1991, 127 (2): 267-268). It has also been reported that ddI causes pancreatitis (Levin, TL et. Al ., Pediatric Radiology, 1997, 27 (2): 189-191).

Therefore, since AIDS therapeutic drugs used to date have many side effects, development of various therapeutic drugs to replace them is urgently required. In line with this, the development of AIDS therapeutic drugs using natural products has recently been activated. For example, gossypol and its synthetic derivatives (Prusoff, W. et. Al ., Pharmacology & Therapeutics, 1993, 60 (2): 315-329), the marine natural alkaloid pyrido [4,3,2] -mn] thiazolo [5,4-b] acridine (pyrido [4,3,2-mn] thiazolo [5,4-b] acridine) (Taraporewala, IB et. al ., J. Med. Chem , 1992, 35 (15): 2744-2752), tannins (Nakashima, H. et. Al ., Antiviral Research, 1992, 18: 91-103), flavonoids (Kusumoto, IT et. Al ., Shoyakugaku Zasshi, 1993, 47: 291-294) have been reported to have HIV-I inhibitory activity. In addition, extensive screening of natural products X to find substances with HIV inhibitory activity is being conducted (Yu et al ., 1998, 29 (4): 338-346; Kusumoto, IT et. Al ., Phytother. Res., 1995. 9: 180-184).

Yin-tree has been called thawing in the oriental medicine, and its bark has been called thawing blood. The name of the tree is Kalopanax pictus, a deciduous tree of 20-25m in height that belongs to the Araliaceae and grows in the mountains of the country. The bark is earthy brown, deeply divided and the leaves are 10-30cm long, lanceolate, with fine teeth on the edge. The flowers are 4-5mm in diameter, light yellow, and the petals and stamens are 4-5 each (Korean Plant Book, Ko Kyung-sik, 1991, 275).
Chemical compounds of saponins include kalopanaxsaponin A (Khorlin A Ya, et al. CA 1964 60: 15964d), kalopanaxsaponin A (Korean Journal of Pharmacognosy, 33 (4), 277-284 (2002)) , Alkaloids, tannins, essential oils (Suprynov NI, et al., CA, 1964, 61: 962 g), and the like, and lignan component lyodendrin (Korean Journal of Pharmacognosy 26 (2) 122-129, 1995) has also been reported. The pharmacological activity of M. bark was methanol extract from M. bark experimentally recovering hepatic dysfunction in rabbits (Korean Journal of Food and Nutrition, 18 (3), 273-278, 1989). Recently, it has been known to have various pharmacological effects such as anti-inflammatory action, renal cortical hormone-like action, tonic action, hypoglycemic action, etc. In addition, leaf and bark extracts inhibit inflammatory edema, Mice fed with fruit extract were predominantly subjected to high sugar loading experiments. In addition, the leaf extract has an inhibitory effect on the central nervous system, and the root extract is reversely stimulated.The saponin of Ringi is similar to Ginseng and is used as a substitute. It has not been reported to do.

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Therefore, the present inventors completed the present invention by preparing an extract from the tree using water or methanol, and confirming that the extract has HIV reverse transcriptase, protease and glucosidase and HIV virus proliferation inhibitory activity.

It is an object of the present invention to provide an extract of Yin-wood extracted using water, alcohol or water and an alcohol mixture, and an AIDS therapeutic agent containing the yin-wood extract as an active ingredient.

In order to achieve the above object, the present invention provides an extract of the yeast tree having HIV reverse transcriptase inhibitory activity, HIV protease inhibitory activity, glucosidase inhibitory activity, and HIV replication inhibitory activity, which is extracted in water, alcohol or aqueous alcohol solution.

In addition, the present invention provides an AIDS therapeutic agent containing the extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides an extract of Uk, which has HIV reverse transcriptase inhibitory activity, HIV protease inhibitory activity, glucosidase inhibitory activity, and HIV replication inhibitory activity, and is extracted with water, alcohol or aqueous alcohol solution.

Extract of the present invention is an extract of the tree using water, alcohol or an aqueous alcohol solution. The alcohol in the above may be ethanol or methanol, preferably methanol. The aqueous alcohol solution may be selected from the group consisting of 10% to 100% ethanol and 10% to 100% methanol. In addition, the tree may be selected from the group consisting of leaves, stems, side bottles and roots of the tree.

In a preferred embodiment of the present invention, extracts 1 to 6 were prepared by extracting reflux using water or methanol as a solvent and then freeze-drying the leaves, stems, or side bottles of the nectar (see Table 1). Specifically, water extract (extract 1) of the leaves of Methanol, methanol extract (extract 2) of the leaves, water extract (extract 3) of the stem, methanol extract (extract 4) of the stem, water extract (extract 5) of the side bottle and side bottles Methanol Extract (Extract 6) was prepared.

In order to determine the effect of the extract of the present invention on the HIV proliferation-related enzyme activity and HIV proliferation, which are the main etiologies of AIDS, AIDS inhibition experiments were performed using extracts 1 to 6 extracted from the present invention. Are slightly different depending on the extraction site and extraction solvent, but most of them have HIV-I reverse transcriptase inhibitory activity (see Table 2), protease inhibitory activity (see Table 3 and FIG. 1), and glucosidase inhibitory activity (see Table 4). And HIV-I proliferation inhibitory activity (see Table 5 and FIG. 2). Therefore, the extract of the present invention, which is a species of retrovirus and is excellent in inhibiting the proliferation of HIV, which is a major etiology of AIDS, can be usefully used to inhibit or treat AIDS.

In addition, the present invention provides an AIDS therapeutic agent containing the extract as an active ingredient.

AIDS therapeutics containing the extract of the present invention as an active ingredient can be administered orally or parenterally during clinical administration and can be used in the form of general pharmaceutical preparations.

That is, the formulation of the present invention can be administered in various oral and parenteral formulations during actual clinical administration, and when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used Is prepared using. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, water Prepared by mixing sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

In the AIDS therapeutic of the present invention, the effective dose of the extract of Yin is 10 to 1000 mg / kg, preferably 10 to 100 mg / kg, and may be administered 1 to 3 times.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

Example 1 Preparation of Extract

The leaves, stems, and side bottles of the nectar were collected, washed, and dried in a well ventilated area. 100 g of water or 100 ml of 100% methanol was added to 5 g of the dried yeast sample, followed by extraction for 3 hours. The extracted solution was concentrated under reduced pressure and lyophilized. In the case of methanol extract, the final concentration of methanol was lyophilized so as not to exceed 2% and used as a material for the activity test. Samples were used after dilution at 100 μg / ml in sterile distilled water or DMSO at the time of activity experiment (Table 1).

Used part Extraction solvent extract leaf water Extract 1 leaf Methanol Extract 2 stem water Extract 3 stem Methanol Extract 4 Side disease water Extract 5 Side disease Methanol Extract 6

<Example 2> Effect of Yin-tree extract on HIV-related enzyme activity

<2-1> Inhibition of HIV-I Reverse Transcriptase Activity

RT detection kit (RT-Detect ^™ kit NEK-070A) (Dupont medical products, Boston, MA to determine whether the extract of the present invention inhibits the activity of HIV reverse transcriptase (abbreviated as "RT"). , USA) was performed by ELOSA (Enzyme Linked Oligonucleotide Sorbent Assay) method according to the manufacturer's policy. The RT enzyme was used to synthesize HIV-I RT (10 U / μl, 100 mM potassium phosphate, pH 7.1, 1 mM dithiothreitol, 50% glycerol) produced by genetic recombination (Dupont medical products, Boston, MA, USA). In enzyme buffer (100 mM Tris-HCl, pH 8.0, 160 mM KCl, 1 mM EDTA, 3 mM dithiothreitol, 0.3% (v / v) Triton X-100, 10% (v / v) glycerol) Diluted to 0.005 U / μl concentration was used. The primer-attached RNA template (Dupont medical products, Boston, MA, USA) and the triphosphate-DNA nucleoside were diluted in the reaction mixture buffer (200 mM Tris-HCl, pH 8.0, 40 mM MgCl ₂ ) and the reaction mixture ( reaction mixture) was prepared. Inhibition of reverse transcriptase activity was carried out by adding 10 µl of the reaction mixture buffer, 20 µl of the reaction mixture, and 5.2 µl of the enzyme buffer, and adding 4 µl (100 µg / ml) of the extract of each tree to a 500 µl test tube (ependorf). Preincubation for minutes. 0.8 µl of RT enzyme was added to the pre-cultured tube, followed by reverse transcription for 1 hour at 37 ° C, and 1 minute reaction at 90 ° C to stop reverse transcription. 10 μl of alkaline solution (5.62% potassium hydroxide, 94.38% water) was added to the finished RNA template and hydrolyzed at 37 ° C. for 15 minutes, followed by a buffer solution (13.6% sodium phosphate, monobasic monohydrate, and 86.4% water). 10 μl was added to neutralize. Transfer 50 μl of the neutralized reaction solution to the streptavidin coated microplate wells for ELOSA reaction, and attach horseradish peroxidase (HRP) -labeled detection probe and biotin-labeled capture probe. 50 μl of the containing ELOSA solution was added and incubated at 37 ° C. for 2 hours (Weber, PC et al., Science, 1989, 243: 85-88). After the reaction was completed, the microplate washer (Immunowash model 1250) (Biorad, Nippon Biorad KK, Tokyo, Japan) was used as a wash solution (2-chloroacetamide solution) to remove remaining reagents or hydrolyzed RNA in the microplate well. Wash three times. 100 μl of a fluorescence detection solution (3,3 ', 5,5'-tetramethylbenzidine substrate solution) was added to the washed microplates for 1 hour at room temperature, followed by a stop solution (2.2% citric acid, 1.5% HCl). 100 μl of 1.5% sulfonic acid, 94.8% water) was added to stop the reaction (Josephy, PD et al ., Journal of Biological Chemistry , 1989, 257: 3669-3675). After the reaction was completed, the absorbance was measured at 450 nm using a spectrophotometer (Biorad 3550-UV) (Biorad, Nippon Biorad KK, Tokyo, Japan), and the RT-activity inhibition rate was calculated according to Equation 1 below. .

As a result, the extracts 1, 2, and 4 of the present invention extracted from the tree were low or almost no reverse transcriptase inhibitory activity, and the extracts 3, 5, and 6 were 82.7%, 94.4%, and 50.1%, respectively, at a concentration of 100 μg / ml. HIV reverse transcriptase inhibitory activity was shown (Table 2).

Extract (100 μg / ml) Reverse transcriptase inhibition (%) Extract 1 20.7 ± 1.7 Extract 2 ND Extract 3 82.7 ± 3.3 Extract 4 ND Extract 5 94.4 ± 0.2 Extract 6 50.1 ± 8.8

In the above, ND indicates that it was not detected.

<2-2> HIV Protease Activity Inhibition

The degree of cleavage of the substrate by the HIV-1 protease produced by genetic recombination was measured by HPLC to analyze the effect of the extract of Yin on the activity of protease, an enzyme involved in viral replication of HIV. The enzyme source used was determined by Kusumoto et al . (Kusumoto, IT et. Al ., Phytother. Res., 1995, 9: 180-184). Ready. Specifically, HIV-I protease was obtained from JM 105 (Kusumoto, IT et. Al ., Phytother. Res., 1995, 9: 180-184), an E. coli strain expressing DNA encoding the HIV-I protease. The obtained HIV-I protease was diluted in a solution of [50 mM NaOAc (pH 5.0), 1 mM EDTA-2Na, 2 mM 2-mercaptoethanol]: glycerol = 75: 25, and was used as a protein institute (Osaka). Oligopeptide described in SEQ ID NO: 1 purchased from Japan, Japan) (His-Lys-Ala-Arg-Val-Leu- (pNO ₂ -Phe) -Glu-Ala-Nle-Ser-NH ₂ (MW 1315)} Diluted at 2 mg / ml in buffer (50 mM NaOAc, pH 5.0) was used as substrate. The oligopeptide recognizes and cleaves the seventh pNO ₂ -Phe to produce a degradation product (pNO ₂ -Phe) -Glu-Ala-Nle-Ser-NH ₂ as set forth in SEQ ID NO: 2. A total of 5 µl of the reaction mixture was prepared by adding 1 µl of buffer, 1.0 µl of substrate, 1 µl of 100 ml of Methanol extract of the present invention, and 2 µl of enzyme solution, respectively. The reaction mixture was reacted at 37 ° C for 1 hour and then 90 ° C. Heating for 60 seconds to stop the HIV-I protease enzyme reaction. The reaction mixture in which the enzyme reaction was stopped was diluted with 35 µl of distilled water and analyzed by HPLC. For HPLC analysis, the column was a LiChrospher 100 RP-18 column (Merck, Darmstadt, FRG) with a column size of 250 × 4 mm, and the solvent was 0.1% TFA and acetonitrile (20% -50% concentration). Gradient), the flow rate was 1.0 ml / min, and the analysis was set at a UV wavelength of 280 nm, and the HPLC apparatus (System controller: Shimadzu SCL-6B, Pump: Shimadzu LC-9A, Detector: Shimadzu SPD-6A (UV spectrophotometric) was used. detector), Recorder & Integrator: Shimadzu C-R6A Chromatopac).

As a result, all of the extracts of the present invention had protease inhibitory activity, of which the protease inhibitory activity of extract 5 was 58.9%, indicating that the extract had the highest inhibitory activity (Table 3 and Fig. 1).

Extract (100 μg / ml) Protease Inhibition (%) Extract 1 14.3 ± 9 Extract 2 7.5 ± 10.1 Extract 3 14.2 ± 14.7 Extract 4 16.4 ± 5.4 Extract 5 58.9 ± 7.4 Extract 6 24.7 ± 14.2

<2-3> Glucosidase Activity Inhibition

Activity of α-glucosidase cleaving ρ-nitrophenyl-α-D-glucoside, a substrate, to confirm whether the extract of the present invention inhibits glucosidase activity The degree was measured by spectroscopy. Α-glucosidase was used for assaying glucosidase activity in a buffer solution (10 mM sodium phosphate, pH 7.0, α-glucosidase (Toyobo Company, Osaka, Japan) obtained from Saccharomyces sp. Diluted to a concentration of 0.5 U / ml in 20% glycerol), and the substrate cleaved by α-glucosidase was ρ-nitrophenyl-α-D-glucopyranoside (ρ-nitrophenyl-α-D-glucopyranoside) ( Nacalai Tesque Inc., Osaka, Japan) was used by diluting to 10 mM concentration in sterile distilled water. 50 μl of the buffer solution (100 mM sodium phosphate, pH 7.0), 100 μl of substrate solution, and 20 μl (100 μg / ml) of the extract of Mt. were prepared to prepare an α-glucosidase reaction solution, which was preliminarily reacted at 37 ° C. for 5 minutes. After 30 μl of α-glucosidase was added, the enzyme was reacted at 37 ° C. for 10 minutes. After stopping the reaction by adding 140 µl of the enzyme stop solution (0.2 M sodium carbonate), the reaction solution was transferred to a microplate well and the absorbance was measured at 405 nm. % inhibition of α-glucosidase activity was calculated according to the following equation.

As a result, the extracts 1, 2, 3, and 5 extracted from the tree did not show glucosidase inhibitory activity, but the extracts 4 and 6 showed 23.9 ± 4.03% and 15.8 ± 4.78%, respectively. It was found that it had a inhibitor of oxidase (Table 4).

Extract (100 μg / ml) Glucosidase Inhibition (%) Extract 1 ND Extract 2 ND Extract 3 ND Extract 4 23.9 ± 4.03 Extract 5 ND Extract 6 15.8 ± 4.78

Example 3 Effect of Yin-tree Extract on HIV Replication

HIV-I replication according to the method of Otake et al . (Otake, T. et. Al ., J. Traditional medicines, 1994, 11: 188-193) to determine whether the extract of the present invention inhibits the replication of HIV-I. Inhibitory activity was measured. The cell used in the experiment was an MT-4 cell line infected with HTLV-1 (Otake, T. et. Al ., J. Traditional medicines, 1994, 11: 188-193), which cells were penicillin G (Banyu Pharmaceutical). , Tokyo, Japan) RPMI provided 100 U / mL, 100 μg / mL streptomycin (Meiji Seika, Tokyo, Japan) and 10% fetal calf serum (FCS) (Flow Laboratories, North Ryde, Australia) Incubation was performed at -1640 medium (Flow Laboratories, Irvine, Scotland) while maintaining 37 ° C with 5% CO ₂ . The virus used HIV-I (strain HTLV-III _B ) obtained from MOLT-4 / HTLV-3III _B cells (Otake, T. et. Al ., J. Traditional medicines, 1994, 11: 188-193). First, MT-1 cells were infected with HIV-1 (HTLV-III _B ) for 1 hour at 50% -tissue culture infective dose (TCID ₅₀ ). Infected cells were resuspended in RPMI-1640 medium at 1 ± 10 ⁵ cell number / ml and put in 200 μl per well in a 96 well culture plate, and at the same time, each of the extracts of the present invention was added at a concentration of 0 to 100 μg / μl. Incubated daily. HIV-1 infected cells and uninfected cells without the addition of the Yin-Thu extract were used as controls. HIV proliferation inhibitors 3'-Azido-3'-deoxythymidine (AZT or zidovudine) and dextran sulfate 8000 (DS8000) (Sigma (St. Louis, Mo.) Cells added with each were used as a positive control group, and were observed by light microscopy after incubation to completely inhibit HIV-1 induced cytopathic effect (CPE, cytopathic effect) on MT-4 cells (IC ₁₀₀ , inhibitory concentration) and The concentration of the extract of cypress that causes cytotoxicity (CC ₀ , cytotoxic concentration) was confirmed, and the extent to which the viability of MT-4 cells was not reduced was determined to be non-cytotoxic.

As a result, the minimum concentration at which the extract of the present invention completely inhibits HIV virus proliferation (cloning) is 100 μg / ml for extract 1, 50 μg / ml for extract 3, 25 μg / ml for extract 5, and 100 for extract 6 Μg / ml (Table 5), HIV virus proliferation inhibitory activity increased with increasing treatment concentration (Fig. 2). However, extract 2 and extract 4 had no virus growth inhibitory activity (Table 5).

In addition, the minimum concentration exhibiting cytotoxicity when treated by cultivating the Yin-wood extract of the present invention was found to be more than 100 ㎍ / ㎖ in extract 1 to extract 6 all showed little cytotoxicity (Table 5).

extract Minimum concentration (µg / ml) that completely inhibits MT-4 / HIV-1 virus replication Minimum concentration that causes cytotoxicity (㎍ / ㎖) Extract 1 100 > 100 Extract 2 NE > 100 Extract 3 50 > 100 Extract 4 NE > 100 Extract 5 25 > 100 Extract 6 100 > 100

NE means no activity.

Example 3 Acute Toxicity Test

Acute toxicity test was conducted using mice and rats to determine whether the extract of the present invention of the present invention is toxic. ICR mice (28 ± 6 g) and Sprague Dawley rats (250 ± 12 g) were used, divided into 5 groups of 15 rats each. After the oral administration to the mouse and rat at the dose of 5, 20, 100 and 500 mg / kg, respectively, the extract of the stem of the present invention was observed for toxicity for 2 weeks. Water was administered as a control.

As a result, none of the five groups died, and no apparent symptoms were found in the control group. Therefore, it was confirmed that the extract of the stem water of the present invention has almost no acute toxicity upon oral administration.

From the above results, the extract of the present invention can be usefully used as an AIDS therapeutic because it has excellent ability to inhibit HIV-related enzyme activity and inhibit HIV replication.

<110> YU, Young-Beob          CHOI, Seung-Hoon          AHN, Kyoo-Seok          SHIM, BUM-SANG <120> Extract of Kalopanax pictus having inhibitory effect on          replication of human immunodificiency virus and therapeutic          agent for AIDS containing the same <130> 3p-08-19 <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> substrate of HIV-1 protease <220> <221> MOD_RES <222> (7) <223> AMIDATION, <220> <221> MOD_RES <222> (10) <223>, Nle <400> 1 His Lys Ala Arg Val Leu Phe Glu Ala Xaa Ser   1 5 10 <210> 2 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> hydrolysate of peptide represented by SEQ. ID. No. One <220> <221> MOD_RES <222> (1) <223> AMIDATION, <220> <221> MOD_RES <222> (4) <223>, Nle <400> 2 Phe Glu Ala Xaa Ser   1 5

Claims (5)

Yin extract extracted by water, alcohol or aqueous alcohol solution to inhibit HIV replication through HIV reverse transcriptase inhibitory activity, HIV protease inhibitory activity. The tree extract according to claim 1, wherein the tree is selected from the group consisting of leaves, stems and side bottles of the tree. The tree extract according to claim 1, wherein the alcohol is ethanol or methanol. The extract of claim 1, wherein the aqueous alcohol solution is selected from the group consisting of 10% to 100% ethanol, 10% to 100% methanol. The AIDS therapeutic containing the extract of claim 1 as an active ingredient.

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