KR101515822B1 - Virus and target cell interaction inhibition - Google Patents

Abstract

The present invention relates to a composition for inhibiting the interaction of a virus with a target cell. The composition comprises a totemron. Tumerone may include ar-tomerone, alpha-toomerone and beta -tumerone. The turmeric extract containing Tumeron also appears to have an inhibitory effect on the interaction of virus and target cells. And a process or a method for extracting turmeron from turmeric. Compositions for inhibiting viral and target cell interactions can be used against H5N1 AI virus infection and from H5N1 AI virus infection. In addition, the composition may be incorporated into chicken feed to defend against viruses such as the H5N1 AI virus.

Description

{VIRUS AND TARGET CELL INTERACTION INHIBITION}

The present invention generally relates to a composition for inhibiting the interaction between a virus and its target cell. More particularly, the present invention relates to a composition comprising a turmerone for inhibiting interaction between an H5N1 AI virus and its target cell.

Curcuma longa (turmeric) is a traditional medicine widely used in Thailand for the treatment of various skin diseases. It has antioxidant activity, anti-protozoan activity, anti-microbial activity, antidote activity, anti- Several activities have been demonstrated such as anti-tumor activity and anti-inflammatory activity.

Three types of Tumeron, ar-Tumerone, alpha-Tumerone and beta-Tumerone are sesquiterpenes, which can be separated from turmeric. ar-Tumelone can be synthesized from a prochiral diol, a styrene derivative and phenylacetic acid without separating from the sulfur.

ar-Tumeron provides a spice flavor of turmeric. It also has anti-thrombogenic, insecticidal and insecticidal properties. In addition, ar-turmerone inhibits the growth of leukemic cell lines.

H5N1, an avian influenza virus strain, is an infectious agent that causes disease in humans and other animal species. Because the H5N1 AI virus is easily transferred between birds, it is a major threat to livestock and farmers. The H5N1 AI has spread to almost all continents, urging the urgent search for disease prevention and defense methods.

The H5N1 AI virus targets the epithelial cell lining of the respiratory tract. The binding of the H5N1 AI virus to the cell surface of the target cell is mediated by the hemagglutinin, a molecule expressed on the surface of the virus. When hemagglutinin binds to a sialic acid molecule on the epithelial cell lining, the epithelial cells introduce the virus into the cell by endocytosis. The virus replicates itself inside the host cell. Then, the virus is cleaved from the cell surface using an enzyme called neuraminidase, and the virus is released from the host cell. This incision may cause the host cell to die because it causes serious damage to the host cell.

Currently, the most commonly used drug for influenza therapy is Oseltamivir (commonly known as Tamiflu). Oxamavivir acts as a neuraminidase inhibitor, controlling the replication of the H5N1 AI virus. However, there are some side effects and problems with this drug. First, the efficacy of Tamiflu against the H5N1 AI virus has not yet been established. In addition, because of the extremely high price of drugs and sudden drug demand, Tamiflu is difficult for anyone to buy easily. Side effects such as nausea, vomiting, dizziness and headache have also been reported in patients treated with tamiflu. Another problem with Tamiflu is that patients treated with this drug develop flu-resistant strains of this drug, alleviating the effects of subsequent drug administration. Other antiviral agents such as amantadine, rimantadine, and zanamavir also have some of the aforementioned problems. Thus, it became clear that new, improved, and widely available drugs are needed. However, due to continued mutagenization of the H5N1 AI virus, the use of such vaccines is limited.

Thus, it is highly desirable to provide an effective and efficient composition that protects host cells from H5N1 AI infection without undue side effects present in existing drugs.

A potential alternative to defend host cells against H5N1 AI infection is to block or down-regulate sialic acid molecules on the epithelial cell lining. In recent years, evidence has been identified that sialic acid can be down-regulated by curcumin. Cucumins are a component of turmeric. In addition, chickens fed with turmeric in livestock feed were defended against avian influenza, but chickens from neighboring farms that did not feed turmeric were found to be infected. Thus, turmeric and its extracts appear to be a very interesting and suitable candidate for drug development for H5N1 AI virus infection.

Jayaprakasha G K, Jagen Mohan Rao L, Sakariah K K, Chemistry and biological activities of C. longa. Trend Food Science and Technology. 2005; 16; 533-548. Singh G, Singh O P and Maurya S. Chemical and biocidal investigations on some essential oils of Indian Curcuma species. Prog. Crystal Growth and Charact. 2002; 45; 75-81. Chatterjee S, Variyar P S, Gholapha S, Padwal-Desai S R, Bongirwar DR. Effect of γ-irradiation on the volatile oil constituents of turmeric (Curcuma longa). Food Res. Int. 2000; 33; 103-106. Leela N K, Tava A, Shafi P M, John S P, Chempakam B. Chemical composition of essential oils of turmeric (Curcuma longa L.). Acta Pharm. 2002; 137-141. Chang, H., Jang, T., Huang, H., Nien, F., Chang, M. J. Supercritical carbon dioxide extraction of turmeric oil from Curcuma longa Linn and purification of turmerones. Separation and Purification Technology. 2006; 47; 119-125. Manzan C C M, Toniolo F S, Bredow E and Povh N P. Extraction of essential oils and pigments from Curcuma longa [L.] by steam distillation and extraction with volatile solvents. J Agric Food Chem. 2003; 51 (23); 6802-6807. Araujo Ca C and Leon L. Biological activities of Curcuma longa L. Mem Inst Oswaldo, Rio de Janeiro. 2001; 96 (5); 723-728. Aratanechemuge Y, Komiya T, Moteki H, Katsuzaki H, Imai K and Hibasami. Selective induction of apoptosis by ar-turmerone isolated from Turmeric (curcuma longa L) in two human leukemia cell lines, but not in human stomach cancer cell line. Int J Mol Med. 2002; 9; 481-484. On the other hand, the genetically diabetic KK-Ay mice (Hypoglycemic effect of turmeric (Curcuma longa L.) on genetically diabetic mice. Biol. Pharm. Bull. 2005; 28 (5); 937-939. Lee H. S. Antiplatelet property of Curcuma longa L. rhizome-derived ar-turmerone. Biores. Tech, 2006; 97; 1372-1376. Pirrung M C, Moreheade T Jr. A sesquidecade of sesquiterpenes; Total Synthesis, 1980-1994. Part A: Acyclic and Monocyclic Sesquiterpens. In The Total Synthesis of Natural Products; Goldsmith D, Ed .; John Wisley: New York. 1997; 10; 29-44. Vyvyan J R, Loitz C, Looper R E, Matting C S, Peterson EA, Staben S T. Synthesis of Aromatic Bisabolene Natural Products via Palladium-Catalyzed Cross-Couplings of Organozinc Reagents. J Org Chem. 2004; 69; 2461-2468. Shishido K and Bando T. Lipase-mediated asymmetric acetylation of prochiral diols directed toward total syntheses of biologically active molecules. J. Mol. Cat. B: Enzymatic. 1998; 5; 183-186. Total asymmetric synthesis of (7S, 9R) - (+) - bisacumol. Tetrahedron: Asymmetry, 2003; 14; 75-78. Mori K. Synthesis of (R) -ar-turmerone and its conversion to (R) -ar-himachalene, a pheromone component of the flea beetle: (R) -ar-himachalene is dextrorotatory in hexane, while levorotatory in chloroform. Tetrahedron: Asymmetry, 2005; 16; 685-692. (Homoptera: Delphacidae) and Plutella xyostella (Lepidoptera: Yponomeutidae) were investigated. The isolates were identified as Curcuma longa L. rhizome against Nilaparvata lugens (Homoptera: Delphacidae) and Plutella xyostella (Lepidoptera: Yponomeutidae). J Asia-Pacific Entomol. 2001; 4; 181-185. Su H C F, Horvat R and Jilani G. Isolation purification, and characterization of insect repellents from Curcuma longa L. J. Agric. Food Chem. 1982; 30; 290-292. Centers for Disease Control and Prevention (CDC). Update: Influenza activity - United States and worldwide, 2006-07 season, and composition of the 2007-08 influenza vaccine. MMWR Morb Mortal Wkly Rep. 2007; 56 (31); 789-94. Hong Kong / 156/97 (H5N1) and A / Hong Kong / 1074 (1984). The neuraminidase inhibitor GS4104 (oseltamivir phosphate) is efficacious against A / Hong Kong / 156 / / 99 (H9N2) influenza viruses. Antiviral Res. 2000; 48 (2); 101-15. E. curcumin effects on sialic acid levels and sialidase activity in Ehrlich ascites tumor bearing mice. Ozen, N. Uslu, E. Ozen, M. Aydin, S. Altug, T. Belce, A, Kokoglu. Tohoku J. Exp. Med. 2002; 197: 221-7. Swayne, DE, Suaze, DL, Schultz-Cherry, S, Tumpey, TM, King, DM, Nakaya, T, Palese, P, Garcia-Satre, A. Recombinant Paramyxovirus Type-1Aan Influenza-H7 Virus as a Vaccine for Protection of Chicken Against Influenza and New Castle Disease. Avian Dis. 2003; 47: 1047-50.

A first aspect of the present invention provides a composition comprising a tomerone for inhibiting interaction between a virus and a target cell.

A second aspect of the invention provides a composition comprising a tomerone and one or more pharmaceutically acceptable carriers to combat H5N1 AI virus infection.

A third aspect of the present invention provides a composition for protecting against H5N1 AI virus infection, comprising a tomerone and one or more pharmaceutically acceptable carriers.

A fourth aspect of the present invention provides a composition comprising a turmeric extract containing Turmerone, for inhibiting interaction between a virus and a target cell.

A fifth aspect of the present invention provides a composition for protecting chicken from viruses, comprising a tomerone and a chicken feed.

Hereinafter, the present invention will be described with reference to the accompanying drawings.
1A shows a structural formula of ar-tomerone that can be used in one embodiment of the present invention.
Figure 1B shows the structure of alpha -tumerone that can be used in embodiments of the present invention.
Fig. 1c shows a structural formula of? -Tumerone replacing the ar-tomerone of Fig. 1a, which can be used in embodiments of the present invention.
Figure 2 shows a structural formula of curcumin according to an embodiment of the present invention.

Hereinafter, the description of the present invention is limited to the composition for inhibiting viruses, the method for producing the composition, and the use of the composition for the sake of brevity and clarity. It is not intended, however, to exclude other applications that would require basic principles, such as general operational features, functional features, or performance features, in various embodiments of the present invention, from various implementations of the present invention.

Exemplary embodiments of the invention include a composition that inhibits the interaction between a virus and a target cell. Each composition provided by the present invention comprises at least a turmeric. Preferably, the tomerone comprises a turmeric extract comprising ar-tomerone (as shown in Figure 1a), alpha -tumerone (as shown in Figure 1b), beta -tumerone (as shown in Figure 1c) . In another embodiment, the compositions provided herein comprise a tomerone and a pharmaceutically active carrier.

Tumelone can be isolated from some pharmaceutical plants, such as turmeric, synthesized from starting materials, or purchased commercially. Tumelen can also be extracted from turmeric using water, hexane, ethyl acetate, ethanol, methanol, chloroform, dichloromethane, basic water or acidic water as a solvent. An example of extracting tetramone from turmeric using hexane, ethyl acetate and ethanol will be described below.

Plant material

To isolate turmeron from turmeric, turmeric ( Curcuma longa L., Zingiberaceae) were collected in Kanchanaburi. The evidence sample BK 63868 was deposited with the Bangkok Plant Specimens of the Ministry of Agriculture, Bangkok Thailand.

turmeric  Preparation of extract

Hexane turmeric  extract:

To obtain a hexane turmeric extract, dry rhizome (1.15 kg) of turmeric was extracted with hexane by a Soxhlet extractor for 8 hours. This hexane solution was concentrated to dryness in a rotary evaporator to give the hexane turmeric extract as a yellow oil (36.88 g, 3.21% dry weight).

Ethyl acetate turmeric  extract:

Ethyl acetate To obtain the turmeric extract, dry rhizome (10 g) of turmeric was extracted with ethyl acetate by a Soxhlet extractor for 8 hours. The solution was concentrated in vacuo and dried to give ethyl acetate turmeric extract as dark red gum (2.69 g, 26.9% dry weight).

70% ethanol turmeric  extract:

To obtain an ethanol turmeric extract, the dry rhizome (10 g) of turmeric was extracted with 70% ethanol by a Soxhlet extractor for 8 hours. The solution was concentrated in vacuo and dried to give 70% ethanol turmeric extract as dark red gum (3.29 g, 32.9% dry weight).

ethanol turmeric  extract:

To obtain an ethanol turmeric extract, the dry rhizome (1.15 kg) of turmeric was extracted with ethanol by a Soxhlet extractor for 8 hours. The solution was concentrated in vacuo and dried to give an ethanol turmeric extract as dark red gum (230 g, 20% dry weight).

Smoke ( Macerated ) ethanol turmeric  extract:

To obtain an ethanolic turmeric extract of burnished ethanol, the dry rhizome of turmeric (10 g) was blanketed with ethanol for 48 hours. The solution was filtered and evaporated to give the crude ethanolic turmeric extract as dark red gum (1.86 g, 18.6% dry weight).

From turbulence ar - Tomeron  detach

To obtain ar-tomerone, ethanol turmeric extract (51.4 g) was dissolved in hexane, dichloromethane / hexane (1-80%), dichloromethane , Eluted with ethyl acetate / dichloromethane (1-40%), ethyl acetate and ethanol, and 10 fractions (F ₁ -F ₁₀ ) were obtained based on thin layer chromatography (TLC) analysis.

Fraction F ₁ (2 g) was purified by column chromatography (silica gel 60 100 g, 70-230 mesh) eluting with a hexane / acetone gradient solvent system and ethanol to give 9 fractions A ₁ -A ₉ ). The double fraction A ₂ (114.4 mg) was further purified by medium pressure liquid chromatography (MPLC). Specifically, it was purified using the system described in U.S. Patent No. 6565745 and eluted with hexane to give ar-tumerone (26 mg, 0.0023% dry weight). Fraction A ₃ (410 mg) was further purified by column chromatography (silica gel 60 60 g, 70-230 mesh) eluting with hexane, hexane / ethyl acetate (1-99%), ethyl acetate, 195 mg, 0.017% dry weight).

From turbulence Coccumine  detach

A fraction F ₃ was recrystallized using ethanol to afford yellow crystals (7.61 g, 0.66% dry weight) of the larger cumin. Fractions F ₄ (4.74 g) also VLC by eluting with 0.1% methanol / dichloromethane, separated in (silica gel, 200 g, 230-400 mesh), larger cumin (670 mg, 0.058% dry wt) was obtained.

Cucuminoid  Extraction and separation

To obtain the cucuminoids, the dried rhizome (1.0 kg) of turmeric was first extracted with hexane for 10 hours in a Soxhlet extractor. This hexane solution was concentrated to dryness in a rotary evaporator to give a hexane turmeric extract as a yellow oil (53.75 g, 5.38% dry weight). It was then extracted with ethyl acetate for 40 hours by a syringe extractor. The ethyl acetate solution was concentrated, filtered and recrystallized with ethanol to give cocuminoids as orange crystals (32.76 g, 3.28% dry weight), mp 176-178 캜. In high pressure liquid chromatography (HPLC) of the orange crystals, 86.5% curcumin (FIG. 2), 13.4% demethoxycucumine and 0.1% bisdemethoxycocumin were identified.

Spectrometer measurement To data  Structure of pure compounds

Infrared (IR) spectra were obtained using a Fourier transform infrared spectrometer. Specifically, a Perkin-Elmer 2000 Fourier transform infrared spectrometer was used. Mass spectra (MS) were recorded on a mass spectrometer such as AGILENT 1100 series LC / MSB TRAP. Nuclear magnetic resonance (NMR) spectra were obtained using an NMR spectrometer system, in particular VARIAN ^UNITY INOVA 400 (proton NMR ( ¹ H NMR): 400 MHz; carbon-13 NMR ( ¹³ C NMR): 100 MHz). The chemical shift (δ) value is expressed in parts per million (ppm), and the coupling constant ( J value) is expressed in Hz. Tetramethylsilane (TMS) was used as an internal standard.

The spectroscopic data obtained for ar-tomerone are as follows: colorless oil; ^{1 H NMR (400 MHz, CDCl} 3): δ1.16 (d, J = 7.0 Hz, 3H), 1.78 (d, J = 1.3Hz, 3H), 2.03 (d, J = 1.2 Hz, 3H), 2.23 (s, 3H), 2.53 ( dd, J = 15.6 and 8.3 Hz, 1H), 2.63 (dd, J = 15.6 and 6.1 Hz, 1H), 3.15-3.25 (m, 1H), 5.95 (sept, J = 1.3 Hz, 1 H) and 7.03 (d, J = 1.6 Hz, 4H); ¹³ C NMR (100 MHz, CDCl ₃ ):? 20.7, 21.0, 22.0, 27.6, 35.3, 52.7, 124.1, 126.6, 129.1, 135.5, 143.7, 155.2 and 199.9; IR (neat): 2962, 2917, 1686, 1621, 1514, 1445, 1373, 1117 and 1011 cm ^-1 ; Electron ionization mass spectroscopy (EI-MS), mass to charge ratio, m / z: 216 (25), 201 (16.7), 132 (16.7), 119 (59.7), 83 (100) and 55 (12.5).

The spectroscopic data obtained for coccumine are as follows: yellow speckle; Melting point: 182-184 占 폚; ^{1 H NMR (400 MHz, CDCl} 3): δ3.87 (s, 6H), 5.72 (s, 1H), 6.40 (d, J = 15.8 Hz, 2H), 6.86 (d, J = 8.1 Hz, 2H) , 6.98 (d, J = 1.8 Hz, 2H), 7.04 (dd, J = 8.1 and 1.8 Hz, 2H) and 7.51 (d, J = 15.8 Hz, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ):? 55.9, 101.1, 109.7, 114.9, 121.7, 122.8, 127.6, 140.5, 146.9, 148.0 and 183.2; IR (KBr): 3406, 1628, 1597, 1508, 1427, 1279, 1156 and 1028 cm ^-1 ; UV / Vis:? _Max 422 nm; Mass spectroscopy (MS) in m / z: 368.8 (100), 284.9 (12.1), 244.9 (26.3), 176.8 (39.5) and 117.1 (9.5).

Pharmacologically acceptable carrier

To prepare a usable pharmaceutical formulation comprising Tumorone, a pharmaceutically acceptable carrier is added. Examples of pharmaceutically acceptable carriers include starch, water, milk, sugar, gelatin, stearic acid or its salts, magnesium stearate, calcium stearate, talc, vegetable fats, vegetable oils, gums, glycols or other excipients do.

Experimental data showing antiviral activity and virus defense properties of the turmeric extract provided by the present invention are shown below.

In vitro Hemyglutination - inhibition analysis

Hemagglutinin analysis was performed to test the virus defense properties and antiviral activity of the turmeric extract. The turmeric extract was diluted with a solvent at a concentration of 0.0001 ng / ml to 100,000 ng / ml. Extracts of each concentration were incubated with 0.5% chicken erythrocyte (rbc) for 30 min followed by addition of H5N1 AI virus-containing emulsion, or incubation with H5N1 AI virus-containing emulsion, followed by addition of 0.5% chicken rbc suspension Respectively. We investigated the hemoglutinization of chicken rbc. Inhibition of hemoglobulinization was detected by the red dot of chicken rbc on the plate bottom. This method is a modification of the method of Swayne et al., 2003.

Reagent preparation

The antiviral and viral defense activity of the turmeric extract was confirmed by in vitro hemagglutination-inhibition assay. The turmeric extracts, including hexane trifoliate extract, ethyl acetate trifoliate extract, 70% ethanol trifoliate extract, ethanol triphosphate extract, ar-tomerone, curcuminoid and pure curcumin, were dissolved in 1% dimethyl (DMSO) at a concentration of 0.05 ng / ml to 500 ng / ml. The test was performed in a 96 well microplate.

The H5N1 AI virus was isolated from infected chickens, inoculated with eggs, and the amniotic fluid from the infected chicken embryos was collected. Verification of H5N1 AI virus strains in amniotic fluid was performed using polymerase chain reaction (PCR).

To determine optimal viral titers, the following steps were performed. The H5N1 AI virus was serially diluted 10 times with 100 mM PBS and mixed with equal volume of 0.5% chicken red blood cells in PBS. The hemogulutylation reaction was observed after incubation at room temperature for 30 minutes. The final concentration of H5N1 AI virus positive for chicken rbc was used in subsequent experiments.

Antiviral activity

To examine the antiviral activity of the turmeric extract, 25 μl of the diluted extract was mixed with the same volume of H5N1 AI virus-containing emulsion for 30 minutes at room temperature, and then 50 μl of 0.5% chicken rbc suspension was added.

A positive control of hemoglobulinization is a mixture of H5N1 AI virus and 0.5% chicken rbc, and the negative control is a mixture of chicken brc and 1% DMSO in PBS.

The inhibition reaction of the turmeric extract was evaluated after 30 minutes. The inhibition of hemoglobulinization was detected as the red spot of chicken rbc on the plate bottom.

The composition, the concentration of the turmeric extract and the number of wells, which show positive amyelinization in the antiviral test, are shown in Table 1 below. Specifically, the number of wells that are positive for the hemoglutinization reaction is inversely proportional to the level of hemogulutylation inhibition by the compound. According to the results in Table 1, no wells showing hemagglutination positive reaction were observed when ar-Turmerone was tested at concentrations of 250 ng / ml, 25 ng / ml, 2.5 ng / ml and 0.25 ng / ml , Which suggests that ar-tomerone inhibits the hemogulutylation of the H5N1 AI virus in antiviral testing. On the other hand, when testing for cucuminoids and pure curcumin, all wells showed a hemagglutination positive reaction, indicating that cucuminoids and pure curcumin did not inhibit hemagglutination, .

Virus defense activity

To investigate the virus defense activity of the turmeric extract, the extract was incubated with chicken rbc for 30 minutes at room temperature, and then 50 占 퐇 of an emulsion containing H5N1 AI virus was added.

A positive control of hemoglobulinization is a mixture of H5N1 AI virus and 0.5% chicken rbc, and the negative control is a mixture of chicken brc and 1% DMSO in PBS.

The inhibition reaction of the turmeric extract was evaluated after 30 minutes. The inhibition of hemoglobulinization was detected as the red spot of chicken rbc on the plate bottom.

The composition showing the hemagglutination positive reaction in the virus defense test, the concentration of the turmeric extract and the number of wells are shown in Tables 2 and 3 below. As with antiviral assays, the number of wells that are positive in the hemogulutylation reaction is inversely proportional to the level of hemagglutination inhibition by the compound. As a result, in the virus defense test, no wells showing a positive hemagglutination test were observed in the four turmeric extracts, i.e., hexane trifoliate extract, ethyl acetate trifoliate extract, 70% ethanol trifoliate extract and ethanol triphosphate extract. The ability of these extracts to protect chicken rbc from the hemagglutination of the H5N1 AI virus after minute incubations has been demonstrated. In contrast, when the cucuminoids and pure cucumins were tested, all wells showed a hemoglobulin positive reaction. Therefore, cucuminoids and cucumins do not have viral defense properties.

The experimental data described above demonstrate antiviral and viral defense capabilities of turmeric extracts including turmerone and turmeric. In the above-described manner, a composition for inhibiting viruses is disclosed. Although only some embodiments of the present invention have been described, it will be apparent to those skilled in the art that numerous modifications and / or modifications can be made, without departing from the scope and spirit of the invention, on the basis of the present invention.

ar - Turmeron , Cookermanino  And Curcumin  Antiviral activity The compounds derived from turmeric The final concentration of the compound derived from turmeric Number of wells showing positive hemagglutination ar-tomerone 2.5 mg / ml (3/3) 250 / / ml (3/3) 25 [mu] g / ml (1/3) 2.5 / / ml (1/3) 250 ng / ml (0/3) 25 ng / ml (0/3) 2.5 ng / ml (0/3) 0.25 ng / ml (0/3) Cookermanino 2.5 mg / ml (3/3) 250 / / ml (3/3) 25 [mu] g / ml (3/3) 2.5 / / ml (3/3) 250 ng / ml (3/3) 25 ng / ml (3/3) 2.5 ng / ml (3/3) 0.25 ng / ml (3/3) Coccumine 2.5 mg / ml (3/3) 250 / / ml (3/3) 25 [mu] g / ml (3/3) 2.5 / / ml (3/3) 250 ng / ml (3/3) 25 ng / ml (3/3) 2.5 ng / ml (3/3) 0.25 ng / ml (3/3)

curcuma  Virus defense effect of extract The compounds derived from turmeric The final concentration of the compound derived from turmeric Number of wells showing positive hemagglutination Hexane trisulfide extract 1 mg / ml (0/3) 100 [mu] g / ml (0/3) 10 [mu] g / ml (0/3) 1 ug / ml (0/3) 100 ng / ml (0/3) 10 ng / ml (0/3) 1 ng / ml (0/3) 0.1 ng / ml (0/3) Ethyl acetate trisulfide extract 2.5 mg / ml (3/3) 250 / / ml (3/3) 25 [mu] g / ml (3/3) 2.5 / / ml (0/3) 250 ng / ml (3/3) 25 ng / ml (3/3) 2.5 ng / ml (3/3) 0.25 ng / ml (3/3) 70% ethanol trioxide extract 2.5 mg / ml (3/3) 250 / / ml (3/3) 25 [mu] g / ml (3/3) 2.5 / / ml (3/3) 250 ng / ml (3/3) 25 ng / ml (0/3) 2.5 ng / ml (3/3) 0.25 ng / ml (3/3)

curcuma  Virus defense effect of extract The compounds derived from turmeric The final concentration of the compound derived from turmeric Number of wells showing positive hemagglutination Ethanol trioxide extract 2.5 mg / ml (3/3) 250 / / ml (0/3) 25 [mu] g / ml (0/3) 2.5 / / ml (0/3) 250 ng / ml (1/3) 25 ng / ml (1/3) 2.5 ng / ml (3/3) 0.25 ng / ml (3/3) Cucuminoid 2.5 mg / ml (3/3) 250 / / ml (3/3) 25 [mu] g / ml (3/3) 2.5 / / ml (3/3) 250 ng / ml (3/3) 25 ng / ml (3/3) 2.5 ng / ml (3/3) 0.25 ng / ml (3/3) Coccumine 2.5 mg / ml (3/3) 250 / / ml (3/3) 25 [mu] g / ml (3/3) 2.5 / / ml (3/3) 250 ng / ml (3/3) 25 ng / ml (3/3) 2.5 ng / ml (3/3) 0.25 ng / ml (3/3)

Claims (18)

A composition for treating or preventing an H5N1 avian influenza (AI) viral infection, comprising a toomerone of one of the following: ar-tomerone,? -tumerone and? -tumerone. A composition for treating or preventing an H5N1 AI viral infection, comprising at least one telomer of one of the following: ar-tomerone, alpha-toomerone and beta -tumerone, and one or more pharmaceutically acceptable carriers. 3. The composition of claim 2 wherein said at least one pharmaceutically acceptable carrier is selected from the group consisting of starch, water, milk, sugar, gelatin, stearic acid or a salt thereof, magnesium stearate, calcium stearate, talc, Glycol. ≪ / RTI > 3. The composition of claim 2, wherein the at least one pharmaceutically acceptable carrier is starch or water. A composition for treating or preventing H5N1 AI viral infectious disease, comprising one to morone of one of the following: ar-tomerone, alpha-toomerone and beta -tumerone, and one or more pharmaceutically acceptable carriers. 6. The composition of claim 5 wherein said at least one pharmaceutically acceptable carrier is selected from the group consisting of starch, water, milk, sugar, gelatin, stearic acid or a salt thereof, magnesium stearate, calcium stearate, talc, Glycol. ≪ / RTI > 6. The composition of claim 5, wherein the at least one pharmaceutically acceptable carrier is starch or water. delete A composition for treating or preventing an H5N1 avian influenza virus infection in a chicken, comprising a turmerone and a chicken feed of one of the following: ar-tomerone, alpha-toomerone and beta -tumerone. delete delete delete delete delete delete delete delete delete

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