CN101883574A - 牙周病菌增殖抑制剂、口腔卫生品及饮食品 - Google Patents
牙周病菌增殖抑制剂、口腔卫生品及饮食品 Download PDFInfo
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Abstract
牙周病菌增殖抑制剂含有使用至少含有机溶剂的提取溶剂而从血桐提取的血桐提取物作为有效成分。或者,牙周病菌增殖抑制剂含有选自宁芬酚-A、宁芬酚-B和宁芬酚-C中的至少一种作为有效成分。牙周病菌增殖抑制剂可配合于例如口腔卫生品和饮食品中使用。
Description
技术领域
本发明涉及牙周病菌增殖抑制剂、以及含有其的口腔卫生品和饮食品。
背景技术
牙周病菌的感染导致发病的牙周病大致分为牙龈炎、牙周炎和咬合性创伤。最近,已知由于牙周病的慢性化而因牙周病菌引起呼吸系统疾病、心脏病、糖尿病、早产等感染疾病。
作为针对牙周病菌的抗菌剂,已知可可泥中所含的以可可成分作为有效成分的抗菌剂(参照专利文献1)。然而,以可可成分作为有效成分的抗菌剂的制造中,需要繁杂的原料前处理,即,为了得到原料可可泥而对可可豆进行烘焙煎制。
另一方面,如专利文献2中所述,已知属于大戟科血桐属(オオバギ属)的血桐(大叶木)的提取物具有抗菌作用。然而,血桐提取物对于牙周病菌的作用至今尚未弄清,在专利文献2中也无任何记载。
专利文献1:国际公开第2003/99304号
专利文献2:日本特开2007-45754号公报
发明内容
本发明是基于发明人等深入研究的结果,即血桐提取物具有抑制牙周病菌增殖的作用而发现的,其目的在于,提供能够不需要繁杂的原料前处理而制造的牙周病菌增殖抑制剂、以及含有其的口腔卫生品和饮食品。
为了实现上述目的,本发明的第1方案中,提供下述牙周病菌增殖抑制剂,其含有使用至少含有机溶剂的提取溶剂而从血桐提取的血桐提取物作为有效成分。
本发明的第2方案中,提供下述牙周病菌增殖抑制剂,其含有选自宁芬酚-A(nymphaeol-A)、宁芬酚-B(nymphaeol-B)和宁芬酚-C(nymphaeol-C)中的至少一种作为有效成分。
本发明的第3方案中,提供下述口腔卫生品,其含有上述第1和第2方案的牙周病菌增殖抑制剂中的任一者。
本发明的第4方案中,提供下述饮食品,其含有上述第1和第2方案的牙周病菌增殖抑制剂中的任一者。
附图说明
[图1]显示对实施例1所述的血桐提取物以高效液相色谱进行分析得到的结果的色谱图。
[图2]显示对实施例2所述的血桐提取物以高效液相色谱进行分析得到的结果的色谱图。
具体实施方式
以下,对本发明的一实施方案详细地进行说明。
本实施方案的牙周病菌增殖抑制剂含有使用至少含有机溶剂的提取溶剂而从血桐提取的血桐提取物作为有效成分。血桐也被称作流血树(Macaranga tanarius),是属于大戟科血桐属的雌雄异株的常绿阔叶树。血桐生长在冲绳(日本南部)、台湾、中国南部、马来半岛、菲律宾、马来西亚、印度尼西亚、泰国等东南亚、澳大利亚北部等地。血桐在树木中成长极快,即使是在荒地废地也能够生长。
作为供于使用提取溶剂的提取中的提取原料,可以使用血桐的所有器官或各器官的构成要素。提取原料可以使用血桐的单独的器官或其构成要素,也可以将血桐的两种以上的器官或其构成要素混合使用。为了提高所得血桐提取物的牙周病菌增殖抑制作用,优选使用含有血桐的果实、种子、花、根、干、茎的前端部、叶身或分泌物(蜡等)的提取原料。茎的前端部含有茎的生长点和叶芽,较叶身柔软,故容易有效地进行提取操作。此外,相对于血桐整体对各器官所占的比例进行比较时,干、根和叶所占的比例高。因此,使用操作容易的血桐叶身作为提取原料,在原料确保容易的方面有利于工业。
提取原料在采集后原样的状态下、或采集后粉碎、破碎或研碎状态下、或在采集和干燥后粉碎、破碎或研碎状态下、或在采集后粉碎、破碎或研碎并进行干燥的状态下,供于提取操作。为了有效地进行提取,优选将提取原料破碎。提取原料的破碎,可以使用例如切割机、剪断机或破碎机进行。经破碎后的提取原料的形状没有特别限定,可以是例如三角形状、四角形状等多角形状。经破碎后的提取原料为多角形状时,优选其一边的长度为1cm左右。提取原料的粉碎可以用例如磨机、破碎机或研磨机进行。提取原料的研碎可以使用例如捏合机或研钵进行。
从提取原料提取血桐提取物之际使用的提取溶剂可以是水与有机溶剂的混合溶剂,也可以是例如,低级醇、二甲基亚砜、乙腈、丙酮、乙酸乙酯、己烷、甘油、丙二醇之类的有机溶剂。作为能够使用的低级醇,可举出例如:甲醇、乙醇、丙醇、异丙醇、丁醇。有机溶剂可以一种溶剂单独使用,也可以多种溶剂混合使用。甘油和丙二醇有时作为例如溶媒或湿润剂而配合于磨牙剂或漱口剂等口腔卫生品中。所以,将甘油或丙二醇用作提取溶剂所得的血桐提取液配合于口腔卫生品中时,可以省略例如作为溶媒或湿润剂而将甘油或丙二醇进一步配合于口腔卫生品中。故,将血桐提取液配合于口腔卫生品中使用时,优选提取溶剂含有甘油或丙二醇。作为提取溶剂使用水与有机溶剂的混合溶剂时,混合溶剂中的有机溶剂的含量优选50体积%以上,更优选80体积%以上。混合溶剂中的有机溶剂的含量为50体积%以上时,可以特别有效地提取血桐中所含的有效成分。有机溶剂优选低级醇,更优选乙醇。
提取溶剂也可溶解有:有机盐、无机盐、缓冲剂、乳化剂、糊精等。
提取通过使提取原料在提取溶剂中浸渍规定时间而进行。提取之际,为了使提取效率提高,也可根据需要进行例如搅拌、加温或其两者。此外,至少为了防止从提取溶剂中提取不需要的杂质,也可以在使用提取溶剂的提取之前,预先将提取原料供于水提取或热水提取而准备除去提取水的提取原料。被推测具有抑制牙周病菌增殖作用的血桐中所含的成分是宁芬酚类。宁芬酚类是水不溶性。通过将血桐在例如开水中煮沸,可将宁芬酚类以外的杂质有效地转移至提取水中而除去。
为了分离提取原料的残渣,从提取原料中提取的血桐提取物被供于固液分离。固液分离可通过例如,过滤、离心分离之类的公知方法进行。固液分离后的液体状血桐提取物也可根据需要进行浓缩。
将液体状血桐提取物中所含的提取溶剂根据需要除去,由此可以得到固体状血桐提取物。从液体状血桐提取物中除去提取溶剂可以通过例如,减压下的加热来进行,也可以通过冷冻干燥来进行。
使用至少含有机溶剂的提取溶剂而从血桐中提取的血桐提取物中,含有选自宁芬酚-A(别名5,7,3’,4’-四羟基-6-香叶基黄烷酮(5,7,3’,4’-tetrahydroxy-6-geranylflavanone))、宁芬酚-B(别名5,7,3’,4’-四羟基-2’-香叶基黄烷酮(5,7,3’,4’-tetrahydroxy-2’-geranylflavanone))、和宁芬酚-C(别名5,7,3’,4’-四羟基-6-(3”’,3”’-二甲基烯丙基)-2’-香叶基黄烷酮(5,7,3’,4’-tetrahydroxy-6-(3”’,3”’-dimethylallyl)-2’-geranylflavanone))中的至少一种。血桐提取物的主要成分是选自宁芬酚-A、宁芬酚-B和宁芬酚-C中的至少一种,即宁芬酚类,其被推测为具有抑制牙周病菌增殖作用的物质。
血桐提取物中进一步含有蜂胶素A(propolin A、别名5,7,3’,4’-四羟基-2’-(7”-羟基-3”,7”-二甲基-2”-辛烯基)-黄烷酮(5,7,3’,4’-tetrahydroxy-2’-(7”-hydroxy-3”,7”-dimethyl-2”-octenyl)-flavanone)。另外,血桐提取物中,作为微量成分,还含有例如:5,7,3’,4’-四羟基-5’-香叶基黄烷酮(5,7,3’,4’-tetrahydroxy-5’-geranylflavanone、别名异宁芬酚B(isonymphaeol-B))、5,7,3’,4’-四羟基-5’-(7”-羟基-3”,7”-二甲基-2”-辛烯基)-黄烷酮(5,7,3’,4’-tetrahydroxy-5’-(7”-hydroxy-3”,7”-dimethyl-2”-octenyl)-flavanone)、5,7,3’,4’-四羟基-6-(7”-羟基-3”,7”-二甲基-2”-辛烯基)-黄烷酮(5,7,3’,4’-tetrahydroxy-6-(7”-hydroxy-3”,7”-dimethyl-2”-octenyl)-flavanone)、5,7,4’-三羟基-3’-(7”-羟基-3”,7”-二甲基-2”-辛烯基)-黄烷酮(5,7,4’-trihydroxy-3’-(7”-hydroxy-3”,7”-dimetyl-2”-octenyl)-flavanone)、5,7,4’-三羟基-3’-香叶基黄烷酮(5,7,4’-trihydroxy-3’-geranylflavanone)。
从血桐各部位提取的提取液之中,从含有蜡的果实提取的提取液特别以高浓度含有宁芬酚A、B、C和异宁芬酚B。
牙周病菌增殖抑制剂在不损害抑制牙周病菌增殖作用的范围内,也可以含有血桐提取物以外的成分。作为可含有在牙周病菌增殖抑制剂中的血桐提取物以外的成分,可举出例如:赋形剂、基质、乳化剂、稳定剂、香料、甜味剂。
牙周病菌增殖抑制剂可以为液状,也可以为固体状。牙周病菌增殖抑制剂的剂型没有特别限定,可以为例如:散剂、粉剂、颗粒剂、片剂、胶囊剂、丸剂、液体剂、注射剂。
牙周病菌的感染导致发病的牙周病大致分为牙龈炎、牙周炎和咬合性创伤。本实施方案的牙周病菌增殖抑制剂由于抑制牙周病菌的增殖,而在牙周病的治疗和预防中使用。
作为牙周病菌,已知属于放线杆菌(Actinobacillus)属、卟啉单胞菌(Prophyromonas)属、普氏菌(Prevotella)属或梭杆菌(Fusobacterium)属的细菌。作为属于放线杆菌属的牙周病菌的例子,可举出例如:伴放线菌放线杆菌(Actinobacillus actinomycetemcomitans)。作为属于卟啉单胞菌属的牙周病菌的例子,可举出例如:牙龈卟啉单胞菌(Prophyromonas gingivalis)、不解糖卟啉单胞菌(Prophyromonasasaccharolytica)、牙髓卟啉单胞菌(Prophyromonas endodontalis)。作为属于普氏菌属的牙周病菌的例子,可举出例如:中间普氏菌(Prevotellaintermedia)、变黑普氏菌(Prevotella nigrescens)、产黑普氏菌(Prevotellamelaninogenica)。作为属于梭杆菌属的牙周病菌的例子,可举出例如:具核梭杆菌(Fusobaeterium nucleatum)、坏死梭杆菌(Fusobacteriumnecrophorum)、舟型梭杆菌AA(Fusobacterium naviforme)。
这些牙周病菌之中,属于放线杆菌属的牙周病菌和属于卟啉单胞菌属的牙周病菌被定为牙周病的主要原因菌。本实施方案的牙周病菌增殖抑制剂通过对属于放线杆菌属的牙周病菌和属于卟啉单胞菌属的牙周病菌中的至少一者的增殖进行抑制,而在牙周病的预防和治疗中发挥优异的效果。
牙周病菌增殖抑制剂被利用作为口腔卫生品、饮食品、医药品、准标准药物等。牙周病菌增殖抑制剂在易于使有效成分接触牙周组织、易于充分发挥有效成分的作用方面,优选被利用作为口腔卫生品或饮食品。
本实施方案的口腔卫生品含有上述牙周病菌增殖抑制剂。作为口腔卫生品的例子,可举出例如:粉磨牙剂、炼磨牙剂、液状磨牙剂、漱口剂、牙龈按摩膏、局部涂布剂、含片剂、口香糖、牙线。含有牙周病菌增殖抑制剂的口腔卫生品可以抑制口腔内的牙周病菌增殖,因此在牙周病的预防和治疗中有效。口腔卫生品中,按固体成分换算,以优选0.001~10质量%、更优选0.01~1质量%的范围含有血桐提取物。按固体成分换算,口腔卫生品中的血桐提取物的含量为0.001质量%以上时,通过口腔卫生品,牙周病菌的增殖特别有效地受到抑制。但是,按固体成分换算,口腔卫生品中的血桐提取物的含量超过10质量%时,得不到与其含量成比例的高牙周病菌增殖抑制效果。口腔卫生品中,除了对应于其形态的基材之外,也可根据需要含有例如乳化剂、溶媒、稳定剂。
本实施方案的饮食品(饮料品和食品)含有上述牙周病菌增殖抑制剂。作为饮料品的例子,可举出例如:碳酸饮料、茶饮料、清凉饮料、酒类、牛奶、咖啡、水果汁、胶体饮料、营养饮料剂。碳酸饮料可以是例如:汽水、鲜柠檬苏打水、可乐。茶饮料可以是例如:绿茶、乌龙茶、红茶。清凉饮料可以是例如:加果汁饮料、矿泉水。酒类可以是例如:啤酒、发泡酒、日本酒、威士忌、烧酒、鸡尾酒。作为食品的例子,可举出例如:酸乳酪、汤、咖喱、润喉糖、糖果、小甜饼、糕饼、日本点心、小吃点心、果汁。
饮食品可以是例如:健康食品、特定保健用食品、健康饮料、营养辅助食品。按固体成分换算,饮食品中以优选0.001~10质量%、更优选0.01~1质量%的范围含有血桐提取物。按固体成分换算,饮食品中的血桐提取物的含量为0.001质量%以上时,通过饮食品,牙周病菌的增殖特别有效地受到抑制。但是,按固体成分换算,饮食品中的血桐提取物的含量超过10质量%时,得不到与其含量成比例的高牙周病菌增殖抑制效果。饮食品中,除了对应于其形态的基材之外,还可根据需要含有例如乳化剂、溶媒、稳定剂。
根据本实施方案,可获得以下的效果。
本实施方案的牙周病菌增殖抑制剂含有血桐提取物作为有效成分。血桐提取物可通过使用至少含有机溶剂的提取溶剂对血桐进行提取,而容易且有效地得到。总之,本实施方案的牙周病菌增殖抑制剂能够不需要繁杂的原料前处理地制造。
此外,由于血桐在树木中生长极快,在荒地废地中也能够生长,故在栽培上不费功夫。并且,血桐提取物由于是植物来源,故安全性高。所以,本实施方案的牙周病菌增殖抑制剂在原料的供给稳定性、生产率、安全性方面均优异。
本实施方案的口腔卫生品和饮食品由于含有上述牙周病菌增殖抑制剂,故具有与牙周病菌增殖抑制剂相同的效果。
前述实施方案可以如下地进行改变。
牙周病菌增殖抑制剂可以代替血桐提取物或者除了血桐提取物之外,还含有例如选自来源于冲绳产蜂胶之类的、血桐提取物以外的宁芬酚-A、宁芬酚-B和宁芬酚-C中的至少一种作为有效成分。
以下,举出实施例对本发明进一步具体地进行说明。
实施例1
<血桐提取物的制备1>
将在冲绳县采集并冷冻的血桐的鲜叶解冻,用剪子剪细。将剪断的鲜叶30g在90体积份的乙醇与10体积份的水构成的混合溶剂100ml中在静置于室温的状态下浸渍2周。之后,对其进行过滤并将滤液作为血桐提取液回收。通过对血桐提取液进行冷冻干燥,从而制备血桐提取液中所含固体成分的粉末,即血桐提取物。粉末状血桐提取物中的宁芬酚-A、宁芬酚-B和宁芬酚-C的总浓度、即宁芬酚类浓度由图1所示的色谱图算出的结果为50质量%,所述图1所示的色谱图通过在下述HPLC条件下分析血桐提取物而得。
HPLC条件
系统:PDA-HPLC系统(岛津制作所)、LC 10ADvp系列、UV;SPD-10Avp、PDA;SPD-M10Avp
柱:Luna C18(2.0×250mm)(岛津GLC)
溶剂:A:水(5%乙酸)、B:乙腈(5%乙酸)
洗脱条件:0-20min
(梯度洗脱;A∶B=80∶20→A∶B=30∶70)
20-50min
(梯度洗脱;A∶B=30∶70→A∶B=0∶100)
50-60min(A∶B=0∶100)
60-75min(A∶B=80∶20)
流速:0.2ml/min
PDA检测:UV190-370nm
UV检测:UV287nm
注入量:20μl
温度:40℃
<血桐提取物针对牙周病菌的抗菌活性试验>
使用接种环将牙周病菌株1(伴放线菌放线杆菌(JCM8577))和牙周病菌株2(牙龈卟啉单胞菌(JCM12257))分别接种于GAM琼脂培养基后,在35℃培养7天。
使用接种环从GAM琼脂培养基采集由培养而增殖的牙周病菌株1的菌落,并将其溶解在生理盐水中,由此制备104cfu/ml的接种菌液1。使用接种环从GAM琼脂培养基采集由培养而增殖的牙周病菌株2的菌落,并将其溶解在生理盐水中,由此制备104cfu/ml的接种菌液2。
将上述“血桐提取物的制备1”中得到的血桐提取物混合于GAM琼脂培养基中,制备平板培养基。将以血桐提取物的终浓度为100ppm的方式制备的平板培养基作为样品平板培养基1、以血桐提取物的终浓度为250ppm的方式制备的平板培养基作为样品平板培养基2、以血桐提取物的终浓度为500ppm的方式制备的平板培养基作为样品平板培养基3。此外,将乙醇混合于GAM琼脂培养基,将以乙醇的终浓度为0.7%的方式制备的平板培养基作为空白平板培养基。
将样品平板培养基1~3和空白平板培养基分别在115℃、15分钟的条件下灭菌后,用接种环将上述接种菌液1和接种菌液2涂布于各平板培养基上。之后,使用三菱气体化学社制造的厌氧系统,将各平板培养基上的牙周病菌株1和牙周病菌株2供于35℃下7天的培养。
培养后,确认平板培养基上的菌落的有无,结果对于牙周病菌株1,在空白平板培养基和样品平板培养基1上观测到菌落,在样品平板培养基2、3上未观察到菌落。此外,对于牙周病菌株2,在空白平板培养基上观察到菌落,但在样品平板培养基1~3上未观察到菌落。总之,确认了牙周病菌株1的生长,在血桐提取物的浓度为250ppm以上(换算为宁芬酚浓度则为125ppm以上)的条件下受到阻碍,牙周病菌株2的生长,在血桐提取物的浓度为100ppm以上(换算为宁芬酚浓度则为50ppm以上)的条件下受到阻碍。由该结果可知,本实施例的血桐提取物は、与抑制牙周病菌株1增殖的效果相比,抑制牙周病菌株2增殖的作用效果更高。
实施例2
<血桐提取物的制备2>
首先,将血桐的剪断鲜叶30g在95℃的水中浸渍30分钟。通过过滤除去水,并将残留的叶在100%乙醇中浸渍3天。之后,对其进行过滤并将滤液作为血桐提取液回收。通过对血桐提取液进行冷冻干燥,从而制备血桐提取液中所含固体成分的粉末,即血桐提取物。粉末状血桐提取物中的宁芬酚-A、宁芬酚-B和宁芬酚-C的总浓度,即宁芬酚类浓度は、由图2所示的色谱图算出的结果为40质量%,所述图2所示的色谱图通过在上述HPLC条件下分析血桐提取液而得。
<血桐提取物针对牙周病菌的抗菌活性试验>
与实施例1相同地实施抗菌活性试验,结果对于实施例2中制备的血桐提取物,也得到与实施例1中制备的血桐提取物相同的结果,确认了实施例1中制备的血桐提取物与实施例2中制备的血桐提取物的抗菌活性相同。
Claims (6)
1.牙周病菌增殖抑制剂,其特征在于,含有使用至少含有机溶剂的提取溶剂而从血桐提取的血桐提取物作为有效成分。
2.牙周病菌增殖抑制剂,其特征在于,含有选自宁芬酚-A、宁芬酚-B和宁芬酚-C中的至少一种作为有效成分。
3.权利要求2所述的牙周病菌增殖抑制剂,其中,前述选自宁芬酚-A、宁芬酚-B和宁芬酚-C的至少一种来源于血桐提取物。
4.权利要求1~3中任一项所述的牙周病菌增殖抑制剂,其中,所述抑制剂具有抑制属于放线杆菌属的牙周病菌和属于卟啉单胞菌属的牙周病菌的至少一者的增殖的作用。
5.口腔卫生品,其特征在于,其含有权利要求1~4中任一项所述的牙周病菌增殖抑制剂。
6.饮食品,其特征在于,其含有权利要求1~4中任一项所述的牙周病菌增殖抑制剂。
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PCT/JP2008/072134 WO2009072594A1 (ja) | 2007-12-05 | 2008-12-05 | 歯周病菌増殖抑制剤、口腔衛生品、及び飲食品 |
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JP5309302B2 (ja) * | 2009-03-04 | 2013-10-09 | ポッカサッポロフード&ビバレッジ株式会社 | 血中脂質上昇抑制剤 |
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CN102302526B (zh) * | 2011-09-19 | 2013-05-22 | 广东瀚森生物药业有限公司 | 血桐提取物的制备方法及其应用 |
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EP2216037A1 (en) | 2010-08-11 |
CN101883574B (zh) | 2013-06-26 |
US20130115175A1 (en) | 2013-05-09 |
CA2705780A1 (en) | 2009-06-11 |
EP2216037A4 (en) | 2011-01-05 |
US20100247455A1 (en) | 2010-09-30 |
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