CN108135184A - 屋尘螨过敏性蛋白质去除剂及其组合物 - Google Patents

屋尘螨过敏性蛋白质去除剂及其组合物 Download PDF

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CN108135184A
CN108135184A CN201580083094.6A CN201580083094A CN108135184A CN 108135184 A CN108135184 A CN 108135184A CN 201580083094 A CN201580083094 A CN 201580083094A CN 108135184 A CN108135184 A CN 108135184A
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dermatophagoides pteronyssinus
composition
remover
allergic proteins
dermatophagoides
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高永律
全光柱
金贤敬
H·伯鲁马尔萨米
K·萨卡拉帕迪安
安龙浚
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(plant) An Dragon Dredge Study
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Abstract

本发明涉及屋尘螨过敏性蛋白质去除剂及其组合物，以天然植物由来精油或其水蒸气蒸馏物作为有效成分，通过有效去除屋尘螨由来的过敏原，可以预防、减轻以及缓和、治疗由这些过敏原引起的多种过敏性疾病，而且没有其他副作用，可以亲环境及有效地使用，可以显著降低由屋尘螨诱发的各种疾病的发病率及普通人的暴露几率。

Description

屋尘螨过敏性蛋白质去除剂及其组合物
技术领域
本发明涉及屋尘螨过敏性蛋白质去除剂及其组合物，具体地说涉及天然植物精油或水蒸气蒸馏物由来的屋尘螨过敏性蛋白质的去除剂及含有该去除剂的屋尘螨过敏性蛋白质的去除用或屋尘螨的灭杀用组合物，该去除剂通过将屋尘螨由来的各种过敏性蛋白质变性及中和来有效去除，而且保持对屋尘螨的灭杀能力。
背景技术
由屋尘螨引起的哮喘、过敏性皮肤炎、鼻炎等各种过敏性疾病的流行和严重是全世界性最严峻的公共健康问题[Arlian,L.G.2002.Arthropod allergens and humanhealth.Annu.Rev.Entomol.47,395433；Bessor,J.C.&G.Pauli.201l.Mite allergens:anoverview.Eur.Ann.Allergy Clin.lmmunol.43:141156.]。屋尘螨是最重要的过敏症状的原因，全世界有6500万名至1亿2千万名受到影响[Colloff,M.J.2009.DustMites.Springer,Dordrecht.]。屋尘螨多生长于角质或皮屑等堆积在人们常活动或生活区域的床垫、家具、地毯等上。所有屋尘螨中最优势种类为美洲尘螨[American house dustmite(Dermatophagoides farinae)]和欧洲尘螨[European house dust mite(Dermatophagoides pteronyssinus)]，认定为职业及室内环境中过敏原(allergen)的主要出处[Bessor,J.C.&G.Pauli.2011.Mite allergens:an overview.Eur.Ann.AllergyClin.lmmunol.43:141156.]。这些过敏原产生于屋尘螨的胃肠管，与尿粪便一起排泄。
在由屋尘螨引起的过敏性疾病中，哮喘由于高死亡率[Mart,H.Q.&N.Ferguson.2009.Life-threatening asthma:focus on lung protection.IntensiveCare Medicine,Springer.pp.372382.]而被认定为最重要的疾病。在全世界，5-10％的成人、10-20％的儿童患有哮喘[UK BIVDA.2002.Diagnostics in Healthcare.The BritishIn Vitro Diagnostics Association.]。有报道全世界人口每百万名就有2至87名死亡(平均死亡22名)。大部分的过敏原是分子量为14-60kDa的蛋白质，最值得重视的是这属于21个屋尘螨过敏原中的I组和II组[Arlian,L.G.&T.A.E.Platts-Mills.200l.The biology ofdust mites and the remediation of mite allergens in allergicdisease.J.Allergy Clin.lmmunol.107:S406S413；Thomas,W.R.,W.A.Smith,B.J.Hales,K.L.Mills&R.M.O 2002.Characterization and immunobiology of house dust miteallergens.lnt.Arch.Allergy lmmunol.129:118.]。
屋尘螨数与过敏原的量成正比，由此疾病诱发危险性也增加。在I组的情况，1g的过敏原的量相当于50个左右的屋尘螨。当过敏原的量为2g时，就会引起致敏症状，当10g以上时，就会发病。因此，2-10g的过敏原量作为哮喘危险标准，这是指每克家庭灰尘中100-500个左右的屋尘螨数(Platts-Mills,T.A.E&M.D.Chapman.1987.Dust mites；Immunology,allergic diseases,and environmental control.J.AllergyClin.lmmunol.80:755757；Korsgaard,J.1983.Mite asthma and residency；a case-control study on the impact of exposure to house dust mites indwellings.Am.Rev.Respir.Dis.128,231235；Platts-Mills,T.A.E.et al.1989.Dustmite allergen and asthma-A worldwide problem.J.Allergy Clin.lmmunol.83:416427.)。在韩国的情况，有报告指出，被褥的灰尘和布沙发中采集的灰尘及地毯灰尘中的螨虫个数和过敏原含量很相似，检测到2g以上的情况为31.2％，检测到10g以上的家庭为10％以上(洪泉洙，李美京，1990.家庭灰尘中屋尘螨的I组过敏原(Group 1allergen)含量调查，过敏10:347348)，可以确认很多人暴露在屋尘螨引起的疾病危险性中。
为了防治屋尘螨，主要开发出并使用林丹(gamma-BHC)，甲基嘧啶磷(pirimiphos-methyl)，苯甲酸苄酯(benzyl benzoate)，二乙基间甲苯甲酰胺(diethyl-m-toluamide)，邻苯二甲酸二丁酯(dibutyl phthalate)等合成制剂(Pollart,S.M.,G.W.Ward,Jr.&T.A.E.Platts-Mills.1987.House dust sensitivity and environmentalcontrol.lmmunol.Allergy Clin.North Am.7:447461)。可知的屋尘螨过敏原有排泄物、脱皮角质、分泌物、细胞等[Tovey,E.R.,M.D.Chapman,&T.A.E.Platts-Mills.1981.Mitefaeces are a major source of house dust allergens.Nature 289:592593；Arlian,L.G.&T.A.E.Platts-Mills.2001.The biology of dust mites and the remediation ofmite allergens in allergic disease.J.Allergy Clin.lmmunol.107:S406S413.]。即使防治活螨虫，含有过敏性蛋白质的死螨虫也以该状态存在数个月[Green,W.F.1984.Abolition of allergens by tannic acid.Lancet 324:160.]。因此，即使用杀螨虫剂防治，也难以摆脱其死体存在的过敏性疾病。
目前为止，能够变性或中和屋尘螨过敏原的药剂有丹宁酸(tannic acid)，其1-3％喷雾剂产品在美国市场销售[Woodfolk,J.A.,M.L.Hayden,J.D.Miller,G.Rose,M.D.Chapman&T.A.E.Platts-Mills.1994.Chemical treatment of carpets to reduceallergen:a detailed study of the effects of tannic acid on indoorallergens.J.Allergy Clin.lmmunol.94:1926.]。这些产品虽然在去除Der p I和Der f I等过敏原方面有效(Ehnert,B.,S.Lau-Schadendorf,A.Weber,P.Buettner,C.Schou&U.Wahn.1992.Reducing domestic exposure to dust mite allergen reducesbronchial hypersensitivity in sensitive children with asthma.J.AllergyClin.lmmunol.90:135138)，但是由于屋尘螨广泛存在，其效果有限[Visitsunthom，N.,V.Chirdjirapong,V.Pootong,O.Jirapongsananuruk,P.Pacham,S.Weeravejsukit,V.Mahakittikun&P.Vichyanond.2010.The accumulation of dust mite allergens onmattresses made of different kinds of materials.Asian Pac.J.Allergylmmunol.28:155161.]。另外，用在布上时，就会因丹宁酸产生斑点，并用在地毯或其他材质时，因丹宁酸而引起变色[Woodfolk，J.A.,M.L.Hayden,N.Couture&T.A.E.Platts-Mills.1995.Chemical treatment of carpets to reduce allergen:comparison of theeffects of tannic acid and other treatments on proteins derived from dustmites and cats.J.Allergy Clin.lmmunol.96:325333.]，为此引起消费者的不满，但是目前为止还没有开发出能够替代丹宁酸的有效性单体或复合化合物或组合物。
关于利用以往植物提取物或植物精油的屋尘螨的灭杀或过敏原非活性化剂的典型以往技术列举如下。
韩国公开专利第10-2003-0015822号(2003.02.25公开)提出含有植物提取精油的螨虫灭杀性组合物和含有单萜化合物的螨虫灭杀性组合物，其中上述植物为选自由白千层(Melaleuca leucadendron)、花旗松(Pseudotsuga menziesii)、古蓬阿魏(Ferulagalbaniflua)、极香天竺葵(Pelargonium ordoratissimum)、锉叶天竺葵(Pelargoniumradens)、头状天竺葵(Pelargonium capitatum)、蜡菊(Helichrysum augustifolium)、岩兰草(Andropogon muricatus)、薰衣草(Lavendula officinalis)、马郁兰(Origanummajorana)、香蜂花(Milissa officinalis)、白树(Melaleuca viridiflora)、罗文沙叶(Ravensara aromatica)、洋擦木(Sassafras albudum)、万寿菊(Tagetes erecta)、孔雀草(Tagetes patula)以及马鞭草(Verbena officinalis)组成的组中一种以上，上述单萜化合物为选自由香叶醇(geraniol)、芳樟醇(linalool)、(-)-顺式桃金娘烷醇((-)-cis-myrtanol)、反式桃金娘烷醇(myrtanol)、(-)-桃金娘烯醛(myrtenal)、(-)-桃金娘烯醇(myrtenol)、侧柏酮(thujone)、顺式马鞭草烯醇(verbenol)、(-)-马鞭草烯酮(verbenone)、薄荷酮(menthone)以及吲哚啉(indoline)组成的组中一种以上。
但是，上述以往技术仅仅涉及螨虫灭杀用组合物，完全没有提及屋尘螨的过敏性蛋白质的去除。
另外，韩国公开专利第10-2005-0084666号(2005.08.26公开)公开了灰尘螨排泄物的中和方法及组合物，其中，将由水溶性植物提取物组成的植物性提取物水溶液与螨虫排泄物接触，上述水溶性植物提取物为选自苹果绿茶、特选维司力农(Arkin special)、特选山金车(Arnica special)、鳄梨、鳄梨GW、黑加仑(Black Currant)绿茶、玫瑰(CabbageRose)、猫爪草(Cat's Claw)、红花大果油茶(Cemila Oleifera)、积雪草(Centella)、蔓藤越桔绿茶、蒲公英、藤黄(Garcinia)、葡萄籽、葡萄柚绿茶、绿茶、绿茶浓缩物、绿茶HS、六种植物萃取液(Hexaplant Richter)、特选木槿(special)、天胡荽(Hydrocotyle)GR、薰衣草、七叶树、奶蓟草、橙子绿茶、山金车萃取液(Phytexcell Arnica)、紫松果菊(PurpleConeflower)、鼠尾草GW、特选鼠尾草、
草本提取物(Sedaplant Richter)、圣约翰草(St.John’s Wort)、金缕梅(Witchhazel)GW、西洋蓍草以及这些组合组成的组。
但是，上述以往技术仅仅提及过敏诱发源的灰尘螨的排泄物的中和，没有提及屋尘螨本身或其死体或部分死体等的过敏诱发源。
发明内容
因此，本发明的第一目的是提供含有植物精油或水蒸气蒸馏物的屋尘螨过敏性蛋白质的去除剂，其具有对屋尘螨的强灭杀能力，而且通过对屋尘螨引起的过敏性蛋白质有效非活性化或变性化而得到强去除力。
本发明的第二目的是提供有效的屋尘螨过敏性蛋白质的去除用组合物，该组合物含有根据上述第一目的所述的屋尘螨过敏性蛋白质的去除剂作为有效成分。
本发明的第三目的是提供有效的屋尘螨灭杀用组合物，该组合物含有根据上述第一目的所述的屋尘螨过敏性蛋白质的去除剂作为有效成分。
根据为顺利完成本发明上述第一目的的本发明的一个优选实施方式，提供屋尘螨过敏性蛋白质的去除剂，其中，含有选自由异硫氰酸丙烯酯(Allyl Isothiocyanate)、茴香醚(Anethole)、β-细辛醚(-Asarone)、樟脑(camphor)、Δ-3-蒈烯(-3-carene)、香茅醇(citronellol)、异丙苯(Cumene)、对异丙基苯甲醛(cuminaldehyde)、香叶醛(Geranial)、异胡薄荷醇(Isopulegol)、薰衣草醇(Lavandulol)、氧化芳樟醇(linalool oxide)、肉豆蔻醚(Myristicin)、柠檬醛(Neral)、橙花醇(nerol)、橙花叔醇(Nerolidol)、紫苏醛(Perillaldehyde)、紫苏醇(Perilla alcohol)、萜品油烯(terpinolene)以及百里酚(Thymol)组成的组中至少一种的植物精油或水蒸气蒸馏物，并将屋尘螨过敏性蛋白质进行非活性化或变形。
另外，根据为顺利完成本发明上述第一目的的本发明的一个优选实施方式，提供屋尘螨过敏性蛋白质的去除剂，其中，含有选自由罗勒(basil,Ocimum basilicum)、酸刺柏(cade,Juniperus oxycedrus)、葛缕子(caraway,Carum carvi)、胡萝卜籽(carrot seed,Daucus carota)、荆芥(catnip,Nepeta cataria)、雪松(cedar,Thuja occidentalis)、香芹(celery,Apium graveolens)、香菜(coriander herb,(Coriandrum sativum)、地中海柏木(cypress,Cupressus sempervirens)、柠檬桉(lemon eucalyptus,Eucalyptuscitriodora)、洋葱(garic,Allium sativum)、杜松果(juniperberry,Juniperuscommunis)、莱姆(lime,Citrus aurantifolia)、柑橘(mandarin,Citrus reticulata)、牛至(oregano,Origanum vulgare)、玫瑰草(palmarosa,Cymbopogon martinii)、唇萼薄荷(pennyroyal,Mentha pulegium)、欧洲赤松(pine,Pinus sylvestris)、鼠尾草(Dalmatiansage,Salvia officinalis)、薄荷(spearmint,Mentha spicata)以及夏季香薄荷(summersavory,Satureja hortensis)组成的组中至少一种的植物的精油或其水蒸气蒸馏物，并将屋尘螨过敏性蛋白质进行非活性化或变形。
在这里，上述屋尘螨过敏性蛋白质的去除剂可以为由植物精油和水蒸气蒸馏物的混合物组成的屋尘螨过敏性蛋白质的去除剂。
根据为顺利完成本发明的上述第二目的至第三目的的本发明的一个优选实施方式，提供屋尘螨过敏性蛋白质的去除用或屋尘螨灭杀用组合物，该组合物含有有效量的上述屋尘螨过敏性蛋白质的去除剂。
上述有效量可以为相对于总重量的0.1至50重量％，特别是0.1至10重量％。
另外，上述组合物可以进一步含有由丹宁酸、杀虫剂、杀螨剂、芳香剂或这些混合物组成的添加剂。
选择性地，上述组合物作为溶剂可以含有乙醇、作为表面活性剂可以含有聚氧乙烯+聚氧丙烯(9:1)苯乙烯化苯醚以及蒸馏水。
另外，选择性地，上述组合物可以含有月桂醇聚氧乙烯醚(polyoxyethylenelauryl ether)、吐温(polysorbate)80、作为溶剂的乙醇以及蒸馏水。
另一方面，上述组合物的剂型可以为气雾剂、喷剂、液剂、悬浊剂、粉剂或胶囊剂。
另外，根据本发明，提供含有上述屋尘螨过敏性蛋白质去除剂的载体(carrier)。
发明效果
根据本发明的屋尘螨过敏性蛋白质的去除剂及其组合物，可以有效去除屋尘螨由来的过敏原，可以预防、减轻以及缓和、治疗由这些过敏原引起的多种过敏性疾病，而且没有其他副作用，因此可以亲环境及有效地使用，为此可以期待由屋尘螨诱发的各种疾病的发病率及普通人暴露在疾病环境的几率显著下降。
附图说明
图1是将与植物精油反应的蛋白质的反应液进行电泳(SDS-PAGE)，将既有蛋白质的图案与反应液的蛋白质变化状态相互比较的图。
图2是将植物精油与人特异性免疫球蛋白E(Immunoglobulin E,IgE)抗体结合反应按斑点(dot-blot)法进行后的相互比较图。
具体实施方式
本发明涉及的屋尘螨过敏性蛋白质的去除剂，含有选自由异硫氰酸丙烯酯(AllylIsothiocyanate)、茴香醚(Anethole)、β-细辛醚(-Asarone)、樟脑(camphor)、Δ-3-蒈烯(-3-carene)、香茅醇(citronellol)、异丙苯(Cumene)、对异丙基苯甲醛(cuminaldehyde)、香叶醛(Geranial)、异胡薄荷醇(Isopulegol)、薰衣草醇(Lavandulol)、氧化芳樟醇(linalool oxide)、肉豆蔻醚(Myristicin)、柠檬醛(Neral)、橙花醇(nerol)、橙花叔醇(Nerolidol)、紫苏醛(Perillaldehyde)、紫苏醇(Perilla alcohol)、萜品油烯(terpinolene)以及百里酚((Thymol)组成的组中至少一种的植物精油或水蒸气蒸馏物，并将屋尘螨过敏性蛋白质进行非活性化或变形。
另外，本发明涉及的屋尘螨过敏性蛋白质的去除剂，含有选自由罗勒油(basil,Ocimum basilicum)、酸刺柏(cade,Juniperus oxycedrus)、葛缕子(caraway,Carumcarvi)、胡萝卜籽(carrot seed,Daucus carota)、荆芥(catnip,Nepeta cataria)、雪松(cedar,Thuja occidentalis)、香芹(celery,Apium graveolens)、香菜(coriander herb,Coriandrum sativum)、地中海柏木(cypress,Cupressus sempervirens)、柠檬桉(lemoneucalyptus,Eucalyptus citriodora)、洋葱(garic,Allium sativum)、杜松果(juniperberry,Juniperus communis)、莱姆(lime,Citrus aurantifolia)、柑橘(mandarin,Citrus reticulata)、牛至(oregano,Origanum vulgare)、玫瑰草(palmarosa,Cymbopogon martinii)、唇萼薄荷(pennyroyal,Mentha pulegium)、欧洲赤松(pine,Pinussylvestris)、鼠尾草(Dalmatian sage,Salvia officinalis)、薄荷(spearmint,Menthaspicata)以及夏季香薄荷(summer savory,Satureja hortensis)组成的组中至少一种的植物的精油或其水蒸气蒸馏物，并将屋尘螨过敏性蛋白质进行非活性化或变形。
本发明利用植物体组织(叶子、根、果实、茎等)由来的精油或水蒸气蒸馏方法得到精油，该精油具有对屋尘螨的灭杀能力，而且通过将螨虫产生的过敏性蛋白质进行非活性化或变形来去除。
上述螨虫是指屋尘螨(house dust mite)，是屋灰尘中发现的所有螨虫的总称，在屋灰尘中发现的螨虫的种类有报告在日本有17科36种，在韩国1967年头一次报告株等以来，到目前为止报告的韩国的屋尘螨达到总12科26种。
本发明的过敏性蛋白质的去除剂，不仅通过非活性化或变形来保持屋尘螨过敏性蛋白质的去除能力，而且具有对屋尘螨的灭杀能力，该去除剂包括：异硫氰酸丙烯酯(AllylIsothiocyanate)、茴香醚(Anethole)、β-细辛醚(-Asarone)、樟脑(camphor)、Δ-3-蒈烯(-3-carene)、香茅醇(citronellol)、异丙苯(Cumene)、对异丙基苯甲醛(cuminaldehyde)、香叶醛(Geranial)、异胡薄荷醇(Isopulegol)、薰衣草醇(Lavandulol)、氧化芳樟醇(linalool oxide)、肉豆蔻醚(Myristicin)、柠檬醛(Neral)、橙花醇(nerol)、橙花叔醇(Nerolidol)、紫苏醛(Perillaldehyde)、紫苏醇(Perilla alcohol)、萜品油烯(terpinolene)以及百里酚(Thymol)或者这些的任意混合精油或其水蒸气蒸馏物。
本发明的过敏性蛋白质的去除剂，不仅通过非活性化或变形来保持屋尘螨过敏性蛋白质的去除能力，而且具有对屋尘螨的灭杀能力，该去除剂可以包括：罗勒(basil,Ocimum basilicum)、酸刺柏(cade,Juniperus oxycedrus)、葛缕子(caraway,Carumcarvi)、胡萝卜(carrot,Daucus carota)、荆芥(catnip,Nepeta cataria)、雪松(cedar,Thuja occidentalis)、香芹(celery,Apium graveolens)、香菜(coriander herb,(Coriandrum sativum)、地中海柏木(cypress,Cupressus sempervirens)、柠檬桉(lemoneucalyptus,Eucalyptus citriodora)、洋葱(garic,Allium sativum)、杜松果(juniperberry,Juniperus communis)、莱姆(lime,Citrus aurantifolia)、柑橘(mandarin,Citrus reticulata)、牛至(oregano,Origanum vulgare)、玫瑰草(palmarosa,Cymbopogon martinii)、唇萼薄荷(pennyroyal,Mentha pulegium)、欧洲赤松(pine,Pinussylvestris)、鼠尾草(Dalmatian sage,Salvia officinalis)、薄荷(spearmint,Menthaspicata)以及夏季香薄荷(summer savory,Satureja hortensis)或者这些的任意植物混合物的精油或其水蒸气蒸馏物。
另外，本发明提供过敏性蛋白质的去除剂组合物，其特征在于，含有有效量的过敏性蛋白质的去除剂，该去除剂具有对上述屋尘螨的灭杀能力。有效量是指将螨虫的量减少至不会引起过敏所需的量且能够去除或变性过敏性蛋白质。优选地，上述有效量可以为相对于总重量份的0.1至50重量％。另外，上述过敏性中和物质之外还可以含有添加剂。当然，作为添加剂可以选择由丹宁酸、杀虫剂、杀螨剂、芳香剂组成的组中一个或一个以上的添加剂。
本发明中，植物组织的精油或水蒸气蒸馏物具有对美洲尘螨和欧洲尘螨的过敏性蛋白质的去除和灭杀能力，该美洲尘螨和欧洲尘螨为屋尘螨种类中全世界上最优势种类。
进一步包括在上述植物精油或水蒸气蒸馏物中添加通常使用的杀虫剂、杀螨剂、芳香剂等在家庭中常用的工业产品、医药外品、医药品的混合剂，此时本发明的植物精油或水蒸气蒸馏物优选含有0.1至50重量％。植物精油、水蒸气蒸馏物或上述混合剂的使用量可以根据屋尘螨的密度、剂型、适用场所及方法等使用环境而调节。
本发明中，植物组织的精油或水蒸气蒸馏物为对屋尘螨由来的主要过敏性蛋白质具有强去除力的植物体。
在含有上述化合物的植物精油或水蒸气蒸馏物或上述化合物中进一步包括添加通常使用的作为屋尘螨中和剂的丹宁酸的混合剂，此时本发明的植物精油、水蒸气蒸馏物、或上述化合物优选含有0.1至50重量％，另外，在该植物精油、水蒸气蒸馏物、或上述化合物中进一步包括添加杀虫剂、杀螨剂、芳香剂等在家庭中常用的工业产品、医药外品、医药品的混合剂，此时本发明的植物精油或水蒸气蒸馏物优选含有0.1至10重量％。植物精油、水蒸气蒸馏物、上述化合物或上述混合剂的使用量可以根据屋尘螨产生的过敏原的量、中和剂剂型、适用场所及方法等使用环境而调节。
另外，本发明提供屋尘螨过敏性蛋白质的去除剂用组合物，该组合物为含有上述化合物单独或两种或其以上的化合物的混合物。
上述屋尘螨灭杀剂用组合物的使用目的为灭杀屋尘螨，其剂型可以制备成气雾剂、喷剂、液剂、悬浊剂、粉剂、胶囊剂等。
上述屋尘螨过敏性蛋白质的去除剂或变性剂用组合物的使用目的为去除屋尘螨过敏性蛋白质，其剂型可以制备成气雾剂、喷剂、液剂、悬浊剂、粉剂、胶囊剂等。
含有上述植物精油、水蒸气蒸馏物、化合物或混合物的屋尘螨过敏性蛋白质的去除剂用组合物对美洲尘螨和欧洲尘螨产生的过敏性蛋白质具有除去效果。作为过敏性疾病诱发源最强的过敏原组主要为分子量为25kDa的1组和分子量为14kDa的2组，它们都具有90％以上的疾病诱发性。
另外，本发明提供含有上述屋尘螨过敏性蛋白质的去除剂的载体(carrier)，上述载体的例没有特别的限制，可以为壁纸、纺织或无纺织物(fabric)、油漆、内部装修材料的涂层等。
下面通过实验例进一步详细说明本发明，但是这仅仅是为了举例说明本发明，并不限定本发明。
阐述一下，在这里省略各精油或水蒸气蒸馏物的实施例编号(numbering)。
实验例1：对屋尘螨的灭杀能力
关于植物精油或水蒸气蒸馏物的美洲尘螨的灭杀效果，通过蒸气中生物鉴定法进行调查。简略地说，将各4050只成虫(710日龄)接种在聚苯乙烯容器(直径5cm，高2cm)底部的圆形的棉片上。在容器盖中央以直径3cm打孔，在其上面放置细铁丝网后各容器上盖住盖子。将溶解在100μl乙醇的试剂利用直径5cm的滤纸处理后，将滤纸放置在铁丝网上，再盖住新盖子。对照区是只进行100μl的乙醇处理。杀虫率是在处理24小时后在20倍解剖显微镜下进行调查，利用末端尖的木头碰到身体或附件时没有动的螨虫认定为死螨虫。
关于上述确认美洲尘螨的灭杀活性的植物精油和水蒸气蒸馏物的结果，整理并示于表1。
表1
[表1]
对美洲尘螨的植物精油的灭杀活性
实验例2：两种精油混合物对屋尘螨的灭杀能力
关于植物精油或水蒸气蒸馏物对美洲尘螨的灭杀效果，以蒸气中生物鉴定法进行调查(0.17mg/cm2)，结果示于表2。
表2
[表2]
实验例3：对屋尘螨的剂型的灭杀能力
关于溶解于聚氧乙烯+聚氧丙烯(9:1)苯乙烯化苯醚[Ethanolpolyoxyethylene+polyoxypropylene(9:1)styrenated phenyl ether]溶液的胡萝卜籽精油剂型和薄荷精油剂型对美洲尘螨的灭杀效果，利用喷雾生物鉴定法进行调查。
结果分别示于下述表3和表4。
简略地说，将实验剂型从10cm高度连续两次喷雾在直径5cm的圆形黑色棉片上，然后干燥5分钟。将已处理的各圆形黑色棉片放置在一次性碗(直径5cm，高1cm)底面后，将各4050只成虫(710日龄)接种在黑色棉片，盖上碗盖子。将作为常用灭杀剂的优选右旋苯醚菊酯(d-phenothrin)或苄氯菊酯(permethrin顺式：反式，25:75)2.5g/l的喷雾剂用作阳性对照群，将聚氧乙烯+聚氧丙烯(9:1)苯乙烯化苯醚(Ethanolpolyoxyethylene+polyoxypropylene(9:1)styrenated phenyl ether)溶液用作阴性对照群。杀虫率是在处理24小时后在20倍解剖显微镜下进行调查，利用末端尖的木头碰到身体或附件时没有动的螨虫认定为死螨虫。
关于上述胡萝卜籽精油剂型和薄荷精油剂型对美洲尘螨的灭杀活性，分别示于表5和表6。
表3
[表3]
1)聚氧乙烯+聚氧丙烯(9:1)苯乙烯化苯醚(Ethanolpolyoxyethylene+polyoxypropylene(9:1)styrenated phenyl ether)。
2)胡萝卜籽精油5，10，20和30g/L。
表4
[表4]
1)聚氧乙烯+聚氧丙烯(9∶1)苯乙烯化苯醚(Ethanolpolyoxyethylene+polyoxypropylene(9∶1)styrenated phenyl ether)。
2)薄荷精油1，2，3和4％。
表5
[表5]
喷雾处理 杀虫率，1)％(标准误差)
胡萝卜籽精油-5 78±5.8c
胡萝卜籽精油-10 92±5.8b
胡萝卜籽精油-20 100a
胡萝卜籽精油-30 100a
右旋苯醚菊酯(d-phenothrin) 8.3±1.7d
1)P＝0.05(Bonferroni method)
表6
[表6]
1)P＝0.05(Bonferroni method)
实验例4：植物提取物对屋尘螨虫体蛋白质的中和能力
将屋尘螨成虫(羽化后第6-8天)利用磨碎装置在0.01M冷磷酸溶液(pH7.2)中磨碎(例如，屋尘螨80mg/4mL磷酸溶液)。将磨碎物在14000g下离心分离30分钟后，浓缩至蛋白质含量为1mg/mL，之后在零下70℃保管。利用Bradford蛋白质分析设备(kit)测定蛋白质含量，将牛血清白蛋白用作蛋白质标准品。
将屋尘螨蛋白质(20mg)与实验化合物(100，250，500mg)混合，在25℃放置1小时。此时，作为阳性对照化合物使用目前用作屋尘螨中和剂的丹宁酸，阴性对照群只进行4.5μl二甲基亚砜的处理。样品与同量的SDS缓冲液混合，加热10分钟，在12％聚丙烯酰胺中进行电泳，然后在120V电泳80分钟后，将凝胶利用0.1％染色液(Coomassie Blue R-250)染色，确认带型。利用图像分析仪确认具有25kDa和14kDa的分子量的屋尘螨的重要过敏原1组和2组蛋白质的变性。
如图1所示，在凝胶上确认的以M表示的泳道(lane)是蛋白质标记，以Ext表示的泳道是仅加载屋尘螨无处理虫体蛋白质的阴性对照群，5号和6号泳道是显示普通活性的情况，1号、2号、3号和4号是显示强活性的情况，以TA表示的泳道是丹宁酸处理区，为阳性对照群。
关于上述确认屋尘螨中和活性的植物精油及其组合物，整理结果分别示于表7和表8。
表7
[表7]
中和活性 中和活性
精油 活性1) 精油 活性1)
薄荷(Spearmint) +++ 洋葱(Garic) ++
罗勒(Basil) +++ 杜松果(Juniperberry) ++
酸刺柏(Cade) +++ 莱姆(Lime) ++
葛缕子(Caraway) ++ 柑橘(Mandarin) ++
胡萝卜(Carrot)籽 +++ 牛至(Oregano) +++
荆芥(Catnip) ++ 玫瑰草(Palmarosa) ++
雪松(Cedar) +++ 唇萼薄荷(Pennyroyal) ++
香芹(Celery) ++ 欧洲赤松(Pine) +++
香菜(Coriander) ++ 鼠尾草(Sage,Dalmatian) ++
地中海柏木(Cypress) +++ 夏季香薄荷(Savory,summer) +++
柠檬桉(Eucalyptus,lemon) +++
1)与阳性对照区的丹宁酸相当或其以上，+++；
相比阳性对照区，显示有些微小的带(band)去除力的情况，++。
表8
[表8]
a+++，与阳性对照区的丹宁酸相当或其以上；
++，相比阳性对照区，显示有些微小带去除力的情况。
另外，为了确认植物精油或水蒸气蒸馏物及其组合物的中和活性，以斑点(dot-blot)方法进行人特异性免疫球蛋白E(Immunoglobulin E,IgE)抗体结合反应。利用Whatman Hybri-dot manifold，将屋尘螨提取物(20μg/200μl)各自处理在PVDF膜上。用0.1％的PBS-T和5％的skim milk进行封闭(blocking)后，将膜与以1:500稀释的过敏性儿童患者的抗血清结合，在4℃彻夜放置。用0.1％的PBS-T洗涤后，将膜与以1:20000稀释的山羊anti-human IgG(H+L)-HRP一起在常温培养1小时。将试验化合物(各500μg)与以1:250稀释的患者的血清进行试验，然后与以1:5000稀释的IgG-HRP conjugate抗体进行培养。用0.1％的PBS-T洗涤后，将印迹(blot)与Westem blotting检测试剂一起培养一分钟，在室温下在X光胶片上曝光1-10分钟。
效力判定中，作为阳性对照群使用目前作为屋尘螨中和剂使用的丹宁酸(tannicacid)。利用图像分析仪测定与IgE反应的点(dot)的情况。将丹宁酸作为阳性对照群。用于上述结果判读的典型实例示于图2。
图2中，C是无处理对照区；T是丹宁酸(Tannic acid)；1是薄荷(Spearmint)精油；2是夏季香薄荷(Savory,summer)精油；3是牛至(Oregano)精油；4是胡萝卜籽(Carrot seed)精油；5是荆芥(Catnip)精油；6是柑橘(Mandarin)精油；7是莱姆(Lime)精油；8是香芹(Celery)精油。
关于上述植物精油及化合物对两种屋尘螨蛋白质的过敏抗原性(allergenicity)降解，分别示于表9和表10。
表9
[表9]
表10
[表10]
实验例5：决明子(cassia)或肉桂(cinnamon)精油及其他精油混合物对屋尘螨蛋 白质的中和能力
根据实验例4的方法，将决明子(cassia)或肉桂(cinnamon)精油及其他精油混合物以2：1的比例混合时，对屋尘螨虫体蛋白质的中和能力示于表11。
表11
[表11]
工业上利用可能性
本发明涉及的屋尘螨过敏性蛋白质的去除剂及其组合物，可以有效使用于预防、减轻以及缓和、治疗由屋尘螨由来的过敏原引起的多种过敏性疾病，可以期待由屋尘螨诱发的发病率显著降低。

Claims (11)

1.一种屋尘螨过敏性蛋白质的去除剂，其中，所述去除剂含有选自由异硫氰酸丙烯酯(Allyl Isothiocyanate)、茴香醚(Anethole)、β-细辛醚(-Asarone)、樟脑(camphor)、Δ-3-蒈烯(-3-carene)、香茅醇(citronellol)、异丙苯(Cumene)、对异丙基苯甲醛(cuminaldehyde)、香叶醛(Geranial)、异胡薄荷醇(Isopulegol)、薰衣草醇(lavandulol)、氧化芳樟醇(linalool oxide)、肉豆蔻醚(Myristicin)、柠檬醛(Neral)、橙花醇(nerol)、橙花叔醇(Nerolidol)、紫苏醛(Perillaldehyde)、紫苏醇(Perilla alcohol)、萜品油烯(terpinolene)以及百里酚((Thymol)组成的组中至少一种的植物精油或水蒸气蒸馏物，并将屋尘螨过敏性蛋白质进行非活性化或变形。
2.一种屋尘螨过敏性蛋白质的去除剂，其中，所述去除剂含有选自由罗勒(basil,Ocimum basilicum)、酸刺柏(cade,Juniperus oxycedrus)、葛缕子(caraway,Carumcarvi)、胡萝卜籽(carrot seed,Daucus carota)、荆芥(catnip,Nepeta cataria)、雪松(cedar,Thuja occidentalis)、香芹(celery,Apium graveolens)、香菜(coriander herb,Coriandrum sativum)、地中海柏木(cypress,Cupressus sempervirens)、柠檬桉(lemoneucalyptus,Eucalyptus citriodora)、洋葱(garic,Allium sativum)、杜松果(juniperberry,Juniperus communis)、莱姆(lime,Citrus aurantifolia)、柑橘(mandarin,Citrus reticulata)、牛至(oregano,Origanum vulgare)、玫瑰草(palmarosa,Cymbopogon martinii)、唇萼薄荷(pennyroyal,Mentha pulegium)、欧洲赤松(pine,Pinussylvestris)、鼠尾草(Dalmatian sage,Salvia officinalis)、薄荷(spearmint,Menthaspicata)以及夏季香薄荷(summer savory,Satureja hortensis)组成的组中至少一种的植物的精油或其水蒸气蒸馏物，并将屋尘螨过敏性蛋白质进行非活性化或变形。
3.根据权利要求1或2所述的屋尘螨过敏性蛋白质的去除剂，其中，所述屋尘螨过敏性蛋白质的去除剂由植物精油和水蒸气蒸馏物的混合物组成。
4.一种屋尘螨过敏性蛋白质的去除用或屋尘螨灭杀用组合物，其中，所述组合物含有有效量的权利要求1或2所述的屋尘螨过敏性蛋白质的去除剂。
5.根据权利要求4所述的屋尘螨过敏性蛋白质的去除用或屋尘螨灭杀用组合物，其中，以总重量为基准，所述有效量为0.1至50重量％。
6.根据权利要求5所述的屋尘螨过敏性蛋白质的去除用或屋尘螨灭杀用组合物，其中，以总重量为基准，所述有效量为0.1至10重量％。
7.根据权利要求6所述的屋尘螨过敏性蛋白质的去除用或屋尘螨灭杀用组合物，其中，所述组合物进一步含有选自由丹宁酸、杀虫剂、杀螨剂以及芳香剂组成的组中至少一种的添加剂。
8.根据权利要求4所述的屋尘螨过敏性蛋白质的去除用或屋尘螨灭杀用组合物，其中，所述组合物作为溶剂含有乙醇、作为表面活性剂含有聚氧乙烯+聚氧丙烯(9:1)苯乙烯化苯醚以及蒸馏水。
9.根据权利要求4所述的屋尘螨过敏性蛋白质的去除用或屋尘螨灭杀用组合物，其中，所述组合物含有月桂醇聚氧乙烯醚(polyoxyethylene lauryl ether)、吐温(polysorbate)80、作为溶剂的乙醇以及蒸馏水。
10.根据权利要求4所述的屋尘螨过敏性蛋白质的去除用或屋尘螨灭杀用组合物，其中，所述组合物的剂型为气雾剂、喷剂、液剂、悬浊剂、粉剂或胶囊剂。
11.一种含有权利要求1或2所述的屋尘螨过敏性蛋白质去除剂的载体(carrier)。
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CN109938054A (zh) * 2019-04-16 2019-06-28 宁波芮颂生物科技有限公司 一种室内尘螨抗过敏喷雾剂的制备方法
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