HOUSE DUST MITE ALLERGY PROTEIN REMOVER, AND COMPOSITION THEREOF

Abstract

The present invention relates to an agent for removing house dust mite allergens and a composition comprising the same. The composition comprises, as an active ingredient, the agent which is composed of a natural plant essential oil or its steam distillate and can effectively remove allergens derived from house dust mites. Thus, the agent and the composition can be used for the prevention, alleviation, mitigation or treatment of various allergic diseases, which can be caused by allergens derived from house dust mites, in an environmentally friendly and efficient manner without causing particular side effects. In addition, the agent and the composition can significantly reduce the prevalence of diseases caused by house dust mites and the frequency of exposure of people to house dust mites.

Claims

1. An inactivator for house dust mite allergen proteins, which has an ability to denature or neutralize house dust mite allergen proteins, the inactivator being composed of at least one compound selected from the group consisting of allyl isothiocyanate, anethole, beta-asarone, camphor, delta-3-carene, citronellol, cumene, cuminaldehyde, geranial, isopulegol, lavandulol, linalool oxide, myristicin, neral, nerol, nerolidol, perillaldehyde, perilla alcohol, and thymol.

2. An inactivator for house dust mite allergen proteins, which has an ability to denature or neutralize house dust mite allergen proteins, the inactivator being composed of either an essential oil or a steam distillate from at least one plant selected from the group consisting of basil (Ocimum basilicum), cade (Juniperus oxycedrus), caraway (Carum carni), carrot seed (Daucus carota), catnip (Nepeta cataria), celery (Apium graveolens), coriander herb (Coriandrum sativum), cypress (Cupressus sempervirens), lemon eucalyptus (Eucalyptus citriodora), garlic (Allium sativum), juniperberry (Juniperus communis), lime (Citrus aurantifolia), mandarin (Citrus reticulata), oregano (Origanum vulgare), palmarosa (Cymbopogon martinii), pennyroyal (Mentha pulegium), pine (Pinus sylvestris), Dalmatian sage (Salvia officinalis), spearmint (Mentha spicata), and summer savory (Satureja hortensis).

3. The inactivator for house dust mite allergen proteins according to claim 2, wherein the inactivator is composed of a mixture of the plant essential oil and the steam distillate.

4. A composition for inactivating house dust mite allergen proteins or killing house dust mites, the composition comprising an effective amount of an activator for denaturating or neutralizing house dust mite allergens proteins, the inactivator being composed of either an essential oil or a steam distillate from at least one plant selected from the group consisting of basil (Ocimum basilicum), cade (Juniperus oxycedrus), caraway (Carum carvi), carrot seed (Daucus carota), catnip (Nepeta cataria), celery (Apium graveolens), coriander herb (Coriandrum sativum), cypress (Cupressus sempervirens), lemon eucalyptus (Eucalyptus citriodora), garlic (Allium sativum), juniperberry (Juniperus communis), lime (Citrus aurantifolia), mandarin (Citrus reticulata), oregano (Origanum vulgare), palmarosa (Cymbopogon martinii), pennyroyal (Mentha pulegium), pine (Pinus sylvestris), Dalmatian sage (Salvia officinalis), spearmint (Mentha spicata), and summer savory (Satureja hortensis).

5. The composition of claim 4, wherein the effective amount is 0.1-50 wt % based on the total weight of the composition.

6. The composition of claim 5, wherein the effective amount is 0.1-10 wt % based on the total weight of the composition.

7. The composition of claim 6, further comprising at least one additive selected from the group consisting of tannic acid, pesticides, acaricides, and aromatics.

8. The composition of claim 4, further comprising ethanol as a solvent, polyoxyethylene+polyoxypropylene (9:1) styrenated phenyl ether as a surfactant, and distilled water.

9. The composition of claim 4, further comprising polyoxyethylene lauryl ether, polysorbate 80, ethanol as a solvent, and distilled water.

10. The composition of claim 4, which is in the form of aerosol, spray, solution, solution, suspension, powder, or capsule.

11. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] FIG. 1 shows the results of SDS-PAGE electrophoresis of reaction products obtained by reactions between plant essential oils and house dust mite proteins, and shows a comparison between the pattern of known proteins and the pattern of proteins of the reaction products.

[0028] FIG. 2 shows the results of a dot-blot immunoassay performed to examine the human-specific IgE reactivity of plant essential oils.

DETAILED DESCRIPTION OF THE INVENTION

[0029] The present invention is directed to an essential oil or an oil obtained by steam distillation from plant tissue (leaf, root, fruit, stem, etc.), which has acaricidal activity against house dust mites and, at the same time, has the ability to inactivate allergen proteins, produced by house dust mites, by denaturation or inactivation.

[0030] As used herein, the term house dust mites are intended to include all mites that are found in house dust. In Japan, 17-36 species of mites found in house dust were reported, and in Korea, 12-26 species of house dust mites have been reported to date since the first report by Joo et al. in 1967.

[0031] The inactivator for house dust mite allergen proteins according to the present invention has the ability to inactivate house dust mite allergen proteins by denaturation or inactivation and, at the same time, has acaricidal activity against house dust mites. This agent according to the present invention is composed of either at least one compound selected from the group consisting of allyl isothiocyanate, anethole, beta-asarone, camphor, delta-3-carene, citronellol, cumene, cuminaldehyde, geranial, isopulegol, lavandulol, linalool oxide, myristicin, neral, nerol, nerolidol, perillaldehyde, perilla alcohol, and thymol.

[0032] In another embodiment, the inactivator for dust mite allergen proteins according to the present invention has the ability to inacivate house dust mite allergen proteins by denaturation or neutralization and, at the same time, has acaricidal activity against house dust mites, and this inactivator may be composed of either an essential oil or a steam distillate from at least one plant selected from the group consisting of basil (Ocimum basilicum), cade (Juniperus oxycedrus), caraway (Carum carvi), carrot (Daucus carota), catnip (Nepeta cataria), celery (Apium graveolens), coriander herb (Coriandrum sativum), cypress (Cupressus sempervirens), lemon eucalyptus (Eucalyptus citriodora), garlic (Allium sativum), juniperberry (Juniperus communis), lime (Citrus aurantifolia), mandarin (Citrus reticulata), oregano (Origanum vulgare), palmarosa (Cymbopogon martinii), pennyroyal (Mentha pulegium), pine, (Pinus sylvestris), Dalmatian sage (Salvia officinalis), spearmint (Mentha spicata), and summer savory (Satureja hortensis).

[0033] The present invention also provides a composition for inactivating house dust mite allergen proteins, comprising an effective amount of the inactivator for removing house dust mites, which has acaricidal activity against house dust mites. As used herein, the term effective amount refers to an amount required to reduce the amount of mites to a level that does not cause allergy and to denature or neutralize allergen proteins. Preferably, the effective amount may be 0.1-50 wt % based on the total weight of the composition. Further, the composition of the present invention may comprise, in addition to the allergen proteins inactivator, an additive. The additive that may be used in the present invention may be one or more selected from the group consisting of tannic acid, pesticides, acaricides, and aromatics.

[0034] The present invention is directed to an essential oil or a steam distillate from plant tissue, which has the ability to inactivate allergen proteins, produced by Dermatophagoides farinae and Dermatophagoides pteronyssinus, which are the most predominant house dust mite species worldwide, and also has acaricidal activity against Dermatophagoides farinae and Dermatophagoides pteronyssinus.

[0035] The composition of the present invention may further comprise, in addition to the plant essential oil or its steam distillate, at least one additive selected from among conventional pesticides, acaricides, aromatics, industrial products, quasi-drugs, medical drugs, etc., which are frequently used at home. In this case, the plant essential oil or its steam distillate according to the present invention is contained in an amount of 0.1-50 wt % based on the total weight of composition of the present invention. It is to be understood that the content of the plant essential oil, the steam distillate or the additive in the composition may vary depending on various conditions, including the density of house dust mites, the formulation of the composition, a place to which the composition is to be applied, a method for application of the composition, etc.

[0036] The present invention is directed to a plant essential oil or a steam distillate from plant tissue, which has a strong ability to inactivate important allergens derived from house dust mites.

[0037] The compound of the present invention may further comprise, in addition to the plant essential oil or its steam distillate or any compound contained in the plant essential oil or steam distillate, tannic acid which is generally used as an agent for neutralizing house dust mite allergens. In this case, the plant essential oil, the steam distillate or the compound is preferably contained in an amount of 0.1-50 wt % based on the total weight of the composition of the present invention. In addition, the composition of the present invention may further comprise, in addition to the plant essential oil or the steam distillate or the compound, at least one additive selected from among conventional pesticides, acaricides, aromatics, industrial products, quasi-drugs, medical drugs, etc., which are frequently used at home. In this case, the plant essential oil or its steam distillate according to the present invention is contained in an amount of 0.1-10 wt % based on the total weight of the present invention. It is to be understood that the content of the plant essential oil, the steam distillate or the compound or the additive in the composition may vary depending on various conditions, including the amount of allergens produced by house dust mites, the formulation of the neutralizing agent, a place to which the composition is to be applied, a method for application of the composition, etc.

[0038] The present invention also provides a composition for inactivating house dust mite allergen proteins, which contains the compound alone or a mixture of two or more of the compounds.

[0039] The composition for killing house dust mites is used for the purpose of killing house dust mites, and may be formulated in the form of aerosol, spray, solution, suspension, powder, capsule, etc.

[0040] The inactivator is used for the purpose of denaturating or neutralizing house dust mite allergen proteins, and may be formulated in the form of aerosol, spray, solution, suspension, powder, capsule, etc.

[0041] The composition for inactivating house dust mite allergen proteins, which contains the above-described plant essential oil, steam distillate, compound or mixture, exhibits the effect of inactivating allergen proteins produced by Dermatophagoides farinae and Dermatophagoides pteronyssinus.

[0042] The most potent allergens that cause allergic diseases are divided mainly into group I having a molecular weight of 25 kDa and group II having a molecular weight of 14 kDa, which all show an incidence of 90% or more.

[0043] The present invention also provides a carrier comprising the above-described inactivator for denaturating or neutralizing house dust mite allergen proteins. Examples of such a carrier include, but are not limited to, wall paper, woven or nonwoven fabric, a coating layer on an interior finishing material, etc.

[0044] Hereinafter, the present invention will be described in further detail with reference to experimental examples. It is to be understood, however, that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0045] In addition, the numbering of Examples of essential oils and their steam distillates will be omitted.

EXPERIMENTAL EXAMPLE 1

Acaricidal Activity against House Dust Mites

[0046] A vapor-phase mortality bioassay was used to evaluate the acaricidal effect of plant essential oils or steam distillates against adult American house dust mites (Dermatophagoides farina), as described previously [Yun, Y. K., H. K. Kim, J. R. Kim, K. Hwang & Y. J. Ahn. 2012. Contact and fumigant toxicity of Armoracia rusticana essential oil, allyl isothiocyanate and related compounds to Dermatophagoides farinae. Pest Manag. Sci. 68: 788794]. Groups of 40-50 adult mites (both sexes, 7-10 days old) were placed separately onto untreated cotton-fabric circles on the bottom section of polystyrene containers (5 cm diameter2 cm), and each container was sealed with the original tight-fitting lid which had a fine wire screen covering a 3 cm diameter central hole. Appropriate amounts of the test materials in 100 L of ethanol were applied to 5 cm diameter black cotton-fabric circles. Each treated fabric circle was placed on top of the wire screen, which prevented direct contact of adult mites with the test material. Each container was sealed with another solid lid to investigate the potential vapor-phase toxicity of the test materials. Control cotton-fabric circles received 100 L of ethanol only. Mortalities were determined 24 h post-treatment under a dissecting microscope (20). A mite was considered dead if its body and appendages did not move when prodded with a fine wooden dowel, as described previously [Yun, Y. K., H. K. Kim, J. R. Kim, K. Hwang & Y. J. Ahn. 2012. Contact and fumigant toxicity of Armoracia rusticana essential oil, allyl isothiocyanate and related compounds to Dermatophagoides farinae. Pest Manag. Sci. 68: 788794]. All bioassays were replicated three times using 40-50 adults per replicate.

[0047] Plant essential oils and their steam distillates, confirmed to have acaricidal activity against Dermatophagoides farina in the above experiments, are summarized in Table 1 below.

TABLE-US-00001 TABLE 1 Acaricidal activities of plant essential oils against Dermatophagoides farina, measured using a vapor-phase mortality bioassay duringa 24 h exposure Mortality % Concentration (standard Essential oil (mg/cm.sup.2) error) Basil (Ocimum basilicum) 0.35 100 Cade (Juniperus oxycedrus) 0.35 90 1.3 Caraway (Carum carvi) 0.35 98 1.7 Carrot seed (Daucus carota) 0.35 100 Catnip (Nepeta cataria) 0.35 84 2.9 Cedar (Thuja occidentalis) 0.35 100 Celery (Apium graveolens) 0.35 100 Coriander herb (Coriandrum sativum) 0.35 95 4.6 Cypress (Cupressus sempervirens) 0.35 100 Lemon eucalyptus (Eucalyptus citriodora) 0.35 100 Garlic (Allium sativum) 0.35 100 Juniperberry (Juniperus communis) 0.35 95 5.8 Lime (Citrus aurantifolia) 0.35 92 1.2 Mandarin (Citrus reticulata) 0.35 100 Oregano (Origanum vulgare) 0.35 100 Palmarosa (Cymbopogon martinii) 0.35 100 Pennyroyal (Mentha pulegium) 0.35 100 Pine (Pinus sylvestris) 0.35 100 Dalmatian sage (Salvia officinalis) 0.35 94 2.3 Spearmint (Mentha spicata) 0.35 100 Summer savory (Satureja hortensis) 0.35 100

EXPERIMENTAL EXAMPLE 2

Acaricidal Activity of Binary Mixtures of Essential Oils against House Dust Mites

[0048] The acaricidal activity of binary mixtures of plant essential oils or steam distillates against Dermatophagoides farina was evaluated using a vapor-phase mortality assay (0.17 mg/cm.sup.2).

TABLE-US-00002 TABLE 2 Essential oil or Concentration Mortality (%, essential oil mixture or mixing ratio standard error) Spearmint 0.17 mg/cm.sup.2 100 Oregano 0.17 mg/cm.sup.2 100 Palmarosa 0.17 mg/cm.sup.2 99 4.6 Pennyroyal 0.17 mg/cm.sup.2 98 2.9 Carrot seed 0.17 mg/cm.sup.2 100 Summer savory 0.17 mg/cm.sup.2 95 1.7 Spearmint + Oregano 1:1 100 2:1 100 1:2 100 Spearmint + Palmarosa 1:1 100 2:1 100 1:2 100 Spearmint + Pennyroyal 1:1 100 2:1 100 1:2 100 Spearmint + carrot seed 1:1 100 2:1 100 1:2 100 Spearmint + Summer 1:1 100 savory 2:1 100 1:2 100 Carrot seed + Oregano 1:1 100 2:1 100 1:2 100 Carrot seed + Palmarosa 1:1 100 2:1 100 1:2 100 Carrot seed + Pennyroyal 1:1 100 2:1 100 1:2 100 Carrot seed + Summer 1:1 100 savory 2:1 100 1:2 100 Summer savory + 1:1 100 Oregano 2:1 100 1:2 100 Summer savory + 1:1 100 Palmarosa 2:1 100 1:2 100 Summer savory + 1:1 100 Pennyroyal 2:1 100 1:2 100 Oregano + Palmarosa 1:1 100 2:1 100 1:2 100 Oregano + Pennyroyal 1:1 100 2:1 100 1:2 100 Palmarosa + Pennyroyal 1:1 100 2:1 100 1:2 100

EXPERIMENTAL EXAMPLE 3

Acaricidal Activity of Formulations against House Dust Mites

[0049] The acaricidal effect of each of carrot seed essential formulations and spearmint essential oil formulations, dissolved in ethanolpolyoxyethylene+polyoxypropylene (9:1) styrenated phenyl ether, was evaluated using a spray bioassay.

[0050] The results of the evaluation are shown in Tables 3 and 4 below.

[0051] In brief, a spray bioassay was used to evaluate the efficacy of four experimental spray formulations against adult American house dust mites (Dermatophagoides farina) [Lee, J. H., J. R. Kim, Y. R. Koh & Y. J. Ahn. 2013. Contact and fumigant toxicity of Pinus densiflora needle hydrodistillate constituents and related compounds and efficacy of spray formulations containing the oil to Dermatophagoides farinae. Pest Manag. Sci. 69: 696702]. Each test solution was sprayed two times successively at 10 cm upwards onto the 5 cm diameter black cotton-fabric circle. After drying for 5 min, each fabric circle was placed onto the bottom section of a disposable Petri dish (5 cm diameter1 cm height). Groups of 40-50 adult mites (both sexes, 7-10 days old) were placed separately onto the treated fabric circles, and each dish was sealed with the original tight-fitting lid. 2.5 g/L of the commercial acaricide d-phenothrin or permethnrin (cis:trans, 25:75) spray served as a positive control. A negative control consisted of a solution of ethanol-polyoxyethylene+polyoxypropylene (9:1) styrenated phenyl ether in distilled water or water. Mortalities were determined 24 h post-treatment under a dissecting microscope (20). A mite was considered dead if its body and appendages did not move when prodded with a fine wooden dowel, as described previously [Yun, Y. K., H. K. Kim, J. R. Kim, K. Hwang & Y. J. Ahn. 2012. Contact and fumigant toxicity of Armoracia rusticana essential oil, allyl isothiocyanate and related compounds to Dermatophagoides farinae. Pest Manag. Sci. 68: 788794.]. Each treatment was replicated three times using 40-50 adults per replicate.

[0052] The acaricidal activities of carrot seed essential oil formulations and spearmint essential oil formulations against Dermatophagoides farina are shown in Tables 5 and 6 below, respectively.

TABLE-US-00003 TABLE 3 Content (%) Spray Carrot seed Distilled formulation essential oil Surfactant .sup.1) Ethanol water Carrot seed 0.5 2 5 92.5 essential oil-5.sup.2) Carrot seed 1 2 5 92 essential oil-10.sup.2) Carrot seed 2 2 5 91 essential oil-20.sup.2) Carrot seed 3 2 5 90 essential oil-30.sup.2) .sup.1) Polyoxyethylene + polyoxypropylene (9:1) styrenated phenyl ether. .sup.2)5, 10, 20 and 30 g/L of carrot seed essential oil.

TABLE-US-00004 TABLE 4 Content (%) Spray Spearmint seed Distilled formulation essential oil Surfactant .sup.1) Ethanol water Spearmint 1 3 5 91 essential oil-1.sup.2) Spearmint 2 3 5 90 essential oil2.sup.2) Spearmint 3 3 5 89 essential oil-3.sup.2) Spearmint 4 3 5 88 essential oil-4.sup.2) .sup.1) Polyoxyethylene + polyoxypropylene (9:1) styrenated phenyl ether. .sup.2)1, 2, 3 and 4% spearmint essential oils.

TABLE-US-00005 TABLE 5 Mortality (%, Spray formulation standard error).sup.1) Carrot seed essential oil-5 78 5.8 c Carrot seed essential oil-10 92 5.8 b Carrot seed essential oil-20 100 a Carrot seed essential oil-30 100 a d-Phenothrin 8.3 1.7 d .sup.1)P = 0.05 (Bonferroni method).

TABLE-US-00006 TABLE 6 Mortality (%, Spray formulation standard error).sup.1) Spearmint essential oil-1 33 3.3 c Spearmint essential oil-2 73 5.8 b Spearmint essential oil-3 97 3.3 a Spearmint essential oil-4 100 a 2.5 g/L of Permethrin) 17.3 3.3 d.sup. (cis:trans, 25:75) .sup.1)P = 0.05 (Bonferroni method).

EXPERIMENTAL EXAMPLE 4

Neutralizing Activity of Plant Extracts against House Dust Mite Proteins

[0053] Adult mites (both sexes, 6-8 days old) were extracted, as described previously [Nakada, S., K. Ito, C. Urata, Y. Sakamoto, M. Hasegawa, T. Nakagawa & T. Miyamoto. 1985. Allergenecity and immunogenicity of house-dust mite (Dermatophagoides farinae) antigens treated with glutaraldehyde. Ann. Allergy 54: 437441; Tovey, E. R. & B. A. Baldo. 1987. Comparison by electroblotting of IgE-binding components in ectracts of house dust mite bodies and spent mite culture. J. Allergy Clin. Immunol. 79: 93102.]. In brief, whole bodies (80 mg fresh weight/4 mL buffer) were homogenized on ice in 0.01 M phosphate-buffered saline (PBS) (pH 7.4) for 15 min. The homogenate was then stirred at 0 C. for 1 hr, centrifuged for 30 min at 14000 g, and concentrated by ultrafiltration to 1 mg/mL of protein.

[0054] The protein content was determined using a Bradford protein assay kit (Pierce, Rockford, Ill.). Bovine serum albumin (Pierce, Rockford, Ill.) was used as the standard. Precision Plus Protein Standards was supplied by Bio-Rad (Hercules, Calif.).

[0055] SDS-PAGE was performed according to the modified method of Laemmli [Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680685]. Based on the preliminary test results, mixtures of mite proteins (20 mg) and each test material (100, 250, and 500 mg) in 4.5 L of dimethyl sulfoxide (DMSO) were incubated for 1 hr at 251 C. Negative controls consisted of 4.5 L of DMSO. Tannic acid or 3% tannic acid solution served as a positive control and was similarly formulated. Samples were mixed with an equal volume of SDS sample buffer (4% SDS, 4% mercaptoethanol, and 100 mM Tris-HCl, pH 8.0), boiled for 10 min, and resolved by electrophoresis on 12% (w/v) polyacrylamide gels by using a Mini-Protean 3 electrophoresis cell (Bio-Rad, Hercules, Calif.). After electrophoresis at 120 V for 80 min, the gels were stained with 0.1% Coomassie brilliant blue R-250 for quantification. The intensity of denaturing activity of each test material toward the two major HDM allergens group 1 and 2 was determined using a Gel Doc XR image analyzer with Quantity One Software (Bio-Rad, Hercules, Calif.) according to the manufacturer's instructions.

[0056] FIG. 1 shows the results of the gel electrophoresis performed as described above. In FIG. 1, lane M represents a protein marker; land Ext represents a negative control loaded only with untreated house dust mite protein; lanes 5 and 6 indicate moderate activity; lanes 1, 2, 3 and 4 indicate strong activity; and lane TA represents a positive control treated with tannic acid.

[0057] Plant essential oils and compounds, confirmed to have neutralizing activity against house dust mites in the above experiments, are summarized in Tables 7 and 8 below, respectively.

TABLE-US-00007 TABLE 7 Neutralizing activity Essential oil Activity.sup.1) Spearmint +++ Basil +++ Cade +++ Caraway ++ Carrot +++ Catnip ++ Cedar +++ Celery ++ Coriander ++ Cypress +++ Lemon +++ Eucalyptus Garlic ++ Juniperberry ++ Lime ++ Mandarin ++ Oregano +++ Palmarosa ++ Pennyroyal ++ Pine +++ Dalmatian Sage ++ Summer Savory +++ .sup.1)+++: comparable to or higher than the activity of the positive control tannic acid; ++: neutralizing activity somewhat lower than that of the positive control.

TABLE-US-00008 TABLE 8 Neutralizing activity Compound Activity .sup.a Allyl isothiocyanate +++ Anethole ++ Beta-asarone ++ Camphor ++ Delta-3-carene +++ Citronellol ++ Cuminaldehyde +++ Cumene ++ Geranial +++ Isopulegol ++ Lavandulol ++ Linalool oxide ++ Myristicin +++ Neral ++ Nerol ++ Nerolidol ++ Thymol +++ Perillaldehyde +++ Perilla alcohol ++ .sup.a +++: comparable to or higher than the activity of the positive control tannic acid; ++: neutralizing activity somewhat lower than that of the positive control.

[0058] In addition, a dot-blot immunoassay for human-specific IgE reactivity was performed, as described previously [Tsai, L. C., Y. C. Sun, P. L. Chao, M. W. Hung, I. C. Kuo, et al. 2000. Protein sequence analysis and mapping of IgE and IgG epitopes of an allergenic 98 kDa Dermatophagoides farinae paramyosin, Der f 11. Allergy 55: 141147]. House dust mite proteins (10 g) were applied to a PVDF membrane using a Hybri-Dot Manifold (Life Technologies, Gaithersburg, Md.) according to the manufacturer's instructions. After blocking with 0.1% PBS-T containing 5% skim milk, the membrane was incubated with 1:500 diluted allergic patient's antiserum at 4 C. overnight. After washing with 0.1% PBS-T three times, the membrane was incubated with 1:20000 diluted goat anti-human IgG (H+L)-HRP and IgE-HRP conjugate (Zymed Laboratories, Sanfrancisco, Calif.) for 1 hr at 251 C. The essential oils and compounds (500 g each) were tested with the patient's serum which was diluted 1:250, and then incubated 1:5000 diluted IgE-HRP conjugate antibodies. After additional washing, as stated previously, the blot was incubated with ECL chemiluminescence reagent for 1 min and exposed to X-ray films for 5-20 min at room temperature. The intensity of the IgE-reacting dot was determined using a Bio-Rad Gel Doc XR image analyzer, as stated previously. Tannic acid served as a standard reference and was similarly formulated.

[0059] The results of the dot-blot immunoassay performed as described above are shown in FIG. 2. The reference numerals used in FIG. 2 indicate the following: C: untreated control; T: tannic acid; 1: spearmint essential oil; 2: Summer savory essential oil; 3: Oregano essential oil; 4: Carrot seed essential oil; 5: Catnip essential oil; 6: Mandarin essential oil; 7: Lime essential oil; and 8: Celery essential oil.

[0060] The inhibitory activities of the above-described plant essential oils and compounds against the allergenicity of two species of house dust mite proteins are shown in Tables 9 and 10 below, respectively.

TABLE-US-00009 TABLE 9 Inhibition of allergenicity Dermatophagoides Dermatophagoides Essential oil farina pteronyssinus Basil 72.6 75.7 Cade 71.5 75.1 Caraway 64.6 68.7 Carrot 72.4 62.2 Catnip 61.1 53.3 Cedar 67.4 66.5 Celery 59.7 52.8 Coriander 61.5 56.4 Cypress 75.4 67.2 Lemon Eucalyptus 68.6 74.2 Garlic 66.5 56.4 Juniperberry 67.7 58.4 Lime 48.3 69.2 Mandarin 57.6 65.3 Oregano 76.5 62.4 Palmarosa 67.5 62.9 Pennyroyal 71.3 57.0 Pine 68.9 71.7 Dalmatian sage 57.8 48.9 Summer savory 75.1 74.7 Spearmint 78.4 80.3 Tannic acid 87.2 81.1

TABLE-US-00010 TABLE 10 Inhibition of allergenicity Dermatophagoides Dermatophagoides Compound farina pteronyssinus Allyl isothiocyanate 82.1 73.0 Anethole 62.3 52.1 Beta-asarone 64.9 68.5 Camphor 78.2 77.9 Delta-3-carene 77.9 78.1 Citronellol 61.2 49.2 Cuminaldehyde 80.4 76.1 Cumene 60.9 47.8 Geranial 72.6 68.1 Isopulegol 76.5 57.4 Lavandulol 74.4 62.7 Linalool oxide 61.5 66.2 Myristicin 76.3 67.0 Neral 77.2 56.5 Nerol 67.9 42.3 Nerolidol 42.3 67.7 Perillaldehyde 79.1 76.4 Perilla alcohol 68.7 56.5 Thymol 79.4 67.9

EXPERIMENTAL EXAMPLE 5

Neutralizing Activity of Mixtures of Cassia or Cinnamon Essential Oil and Other Essential Oils Against House Dust Mite Proteins

[0061] Cassia or Cinnamon essential oil and other essential oils were mixed at a ratio of 2:1. According to the method of Experimental Example 4, the neutralizing activity of the mixtures was measured, and the results of the measurement are shown in Table 11 below.

TABLE-US-00011 TABLE 11 Inhibition of allergenicity Dermatophagoides Dermatophagoides Essential oil mixture farina pteronyssinus Cinnamon + Oregano 85.2 79.6 Cinnamon + Palmarosa 76.3 70.1 Cinnamon + carrot seed 82.5 74.7 Cinnamon + Summer savory 74.4 78.8 Cassia + Oregano 86.9 78.7 Cassia + Palmarosa 79.7 76.4 Cassia + carrot seed 82.3 79.5 Cassia + Summer savory 80.7 79.8

[0062] As described above, the inactivator for denaturating or neutralizing house dust mite allergen proteins and the composition comprising the same according to the present invention can be effectively used for the prevention, alleviation, mitigation or treatment of various allergic diseases that can be caused by allergens derived from house dust mites. In addition, the inactivator and the composition can significantly reduce the prevalence of diseases caused by house dust mites, and can also significantly reduce the frequency of exposure of people to house dust mites.