JPWO2016194692A1 - 抗う蝕剤及び抗う蝕用組成物 - Google Patents
抗う蝕剤及び抗う蝕用組成物 Download PDFInfo
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- JPWO2016194692A1 JPWO2016194692A1 JP2017521832A JP2017521832A JPWO2016194692A1 JP WO2016194692 A1 JPWO2016194692 A1 JP WO2016194692A1 JP 2017521832 A JP2017521832 A JP 2017521832A JP 2017521832 A JP2017521832 A JP 2017521832A JP WO2016194692 A1 JPWO2016194692 A1 JP WO2016194692A1
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- agent
- cariogenic
- periodontal disease
- bifidobacteria
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Abstract
Description
また、歯周病の予防効果又は治療効果が得られ、かつ副作用のない歯周病予防剤又は歯周病治療剤を提供することにある。
1.ビフィズス菌を含む抗う蝕剤。
2.前記ビフィズス菌がビフィドバクテリウム・ビフィダム(Bifidobacterium bifidum)である前記1に記載の抗う蝕剤。
3.前記ビフィズス菌がビフィドバクテリウム・ビフィダムOLB6378菌株(Bifidobacterium bifidum OLB6378、受託番号:NITE BP−31)である前記1又は2に記載の抗う蝕剤。
4.前記ビフィズス菌が加熱処理された菌体である、前記1〜3のいずれか1に記載の抗う蝕剤。
5.一日当たり108個以上の菌数の前記ビフィズス菌を4週間以上連続して適用される前記1〜4のいずれか1に記載の抗う蝕剤。
6.前記1〜5のいずれか1項に記載の抗う蝕剤を含有する抗う蝕用組成物。
7.さらに分散剤を含有する、前記6に記載の抗う蝕用組成物。
8.前記分散剤がデキストリンである、前記7に記載の抗う蝕用組成物。
9.前記抗う蝕用組成物が抗う蝕用食品組成物である前記6〜8のいずれか1に記載の抗う蝕用組成物。
10.前記抗う蝕用組成物が抗う蝕用医薬組成物である前記6〜8のいずれか1に記載の抗う蝕用組成物。
11.前記10に記載の抗う蝕用組成物を含有する経口用製剤。
12.前記6〜10のいずれか1に記載の抗う蝕用組成物と包材を含み、該抗う蝕用組成物が包材内に包装される、包装体。
13.ビフィズス菌を含む歯周病予防剤又は歯周病治療剤。
14.ビフィズス菌を含む抗う蝕剤、歯周病予防剤又は歯周病治療剤。
15.加熱処理されたビフィドバクテリウム・ビフィダムOLB6378菌株(Bifidobacterium bifidum OLB6378、受託番号:NITE BP−31)。
本発明は、ビフィズス菌を含む抗う蝕剤又は抗う蝕用組成物に関する。
また、本発明は、ビフィズス菌を含む歯周病予防剤もしくは歯周病治療剤、又は歯周病予防用組成物もしくは歯周病治療用組成物に関する。
本発明に使用されるビフィズス菌は、ビフィドバクテリウム属に属する細菌であり、その種類や由来に制限がない。具体的には、例えば、ビフィドバクテリウム・ビフィダム(Bifidobacterium bifidum)、ビフィドバクテリウム・ロンガム(Bifidobacterium longum)、ビフィドバクテリウム・ブレーベ(B.breve)、ビフィドバクテリウム・アドレセンティス(B.adolescentis)、ビフィドバクテリウム・インファンティス(B.infantis)、ビフィドバクテリウム・シュードロンガム(B.pseudolongum)、ビフィドバクテリウム・ラクティス(B.lactis)、ビフィドバクテリウム・サーモフィルム(B.thermophilum)等のビフィズス菌が挙げられる。
(1)寄託機関名:独立行政法人製品評価技術基盤機構 特許微生物寄託センター
(2)連絡先:〒292−0818 千葉県木更津市かずさ鎌足2−5−8(現:千葉県木更津市かずさ鎌足2−5−8 122号室)
電話番号0438−20−5580
(3)受託番号:NITE BP−31
(4)識別のための表示:Bifidobacterium bifidum OLB6378
(5)原寄託日:2004年10月26日
(6)ブダペスト条約に基づく寄託への移管日:2006年1月18日
(1)寄託機関名:独立行政法人産業技術総合研究所(現:国立研究開発法人産業技術総合研究所) 特許生物寄託センター
(現寄託機関名:独立行政法人製品評価技術基盤機構 特許生物寄託センター)
(2)連絡先:〒305−8566 茨城県つくば市東1−1−1 中央第6
電話番号029−861−6029
(現連絡先:〒292−0818 千葉県木更津市かずさ鎌足2−5−8 120号室
電話番号0438−20−5910)
(3)受託番号:FERM BP−11242
(4)識別のための表示:Bifidobacterium longum No.7
(5)原寄託日:平成5年4月20日
(6)ブダペスト条約に基づく寄託への移管日:2010年3月2日
(1)寄託機関名:独立行政法人製品評価技術基盤機構 特許微生物寄託センター
(2)連絡先:〒292−0818 千葉県木更津市かずさ鎌足2−5−8(現:千葉県木更津市かずさ鎌足2−5−8 122号室)
電話番号0438−20−5580
(3)受託番号:NITE P−75
(4)識別のための表示:Bifidobacterium longum OLB6290
(5)寄託日:平成17年2月3日
本明細書ではこれらを、「ビフィズス菌処理物」又は「処理物」と略記することがある。
Bifidobacterium bifidum OLB 6378株(受託番号:NITE BP−31)を、表1に示す組成で調製した嫌気EG培地で37℃にて一夜培養した。
a)加熱処理したOLB6378株の凍結乾燥粉末の調製
実施例1で得られたOLB6378株の原菌末(生菌数、3.9×1011cfu/g)180gを、45℃に調整した原料水2800mlに撹拌しながら完全に懸濁させた。その後、撹拌しながら加温して、90℃15分間保持し、冷却した。得られた加熱菌体の懸濁液を凍結乾燥し、加熱処理したOLB6378株の凍結乾燥粉末156gを得た。なお、加熱処理したOLB6378株にはMRS寒天平板培地上でビフィズス菌の生菌は見られなかった。また、得られたOLB6378株の凍結乾燥粉末156g中には、概算して、7.02×1013個の菌数のビフィズス菌が存在する(3.9×1011(cfu/g)×180(g)=7.02×1013cfu)。熱処理された菌体数は、生菌数(cfu)に換算して示されている。
加熱処理したOLB6378株の凍結乾燥粉末120gと顆粒デキストリン(松谷化学社)2880gを均一に混合し、各1gずつ分包し、本発明の剤とした。この本発明の剤には、概算して、1.8×1010個の菌数のビフィズス菌が存在する(7.02×1013(cfu)/156(g)×120/3000(g)=1.8×1010cfu)。熱処理された菌体数は、生菌数(cfu)に換算して示されている。
実施例2の剤の代わりに、顆粒デキストリン(松谷化学社)3000gを、各1gずつ分包し、比較例1のプラセボとした。
試験例1では、実施例2の剤を摂取した被験者群を「本発明の剤摂取群」とし、比較例1のプラセボを摂取した被験者群を「プラセボ摂取群」とした。被験者群は、後述する被験者の選択基準及び除外基準に従い、両群あわせて30名選定した。このうち、本発明の剤摂取群を15名、プラセボ摂取群を15名とした。両群はいずれも一日二回、実施例2の剤又は比較例1のプラセボを摂取した。摂取期間は5週間である。両群の評価は、後述する評価項目に従って行った。
被験者の選択基準は、以下のA)〜E)のすべてに該当するものとした。
A)20歳以上40歳以下の女性
B)通常1日に3回食事を摂取する者
C)FDI歯コード16(上顎右側第1大臼歯)、21(上顎左側中切歯)、24(上顎左側第1小臼歯)、36(下顎左側第1大臼歯)、41(下顎右側中切歯)、44(下顎右側第1小臼歯)の6歯の唇側・頬側面のうち、4歯以上が測定可能な者。前記の6代表歯の代替としてFDI歯コード17,15,11,22,25,37,35,31,42,45が存在する者
D)スクリーニング検査時に、う蝕及び歯周疾患の治療を受けておらず、歯が20本以上存在する者
E)試験の目的・内容について十分な説明を受け、同意能力があり、よく理解した上で自発的に参加を志願し、書面で試験参加に同意した者
被験者の除外基準は、以下のa)〜j)の少なくとも1に該当するものとした。
a)スクリーニング検査時に、う蝕(C3以上)又は重度の歯周疾患と判断された者
b)糖尿病、慢性腎臓病、胃腸障害、肺疾患、悪性腫瘍等に罹患し、薬剤を服用している者
c)他の医薬品や食品の試験等に現在参加中の者、参加の意志のある者
d)5分間の刺激唾液量が3.0ml以下の者
e)刺激唾液のpHが6.2以下の者
f)可撤性義歯を装着している者
g)スクリーニング検査前1ヶ月以内に、洗口剤、抗生物質、抗菌剤を使用した者、及び歯科を受診した者
h)食物アレルギーを有する者
i)妊娠している者、試験期間中に妊娠の意思がある者、授乳中の者
j)その他、試験責任歯科医師が被験者として不適当と判断した者
1.う蝕菌比率
う蝕菌比率は、本発明の剤が抗う蝕効果を有するか否かの指標となるもので、唾液中のレンサ球菌に占めるミュータンス菌数の割合を示す。すなわち、う蝕菌比率は以下の式で表される。
う蝕菌比率=(ミュータンス菌数/レンサ球菌数)×100
歯肉炎指数は、本発明の剤が歯周病予防効果及び歯周病治療効果を有するか否かの指標となるもので、非特許文献1(Periodontal disease in pregnancy. Lee H,Silness J.,Acta Odontologica Scandinavica 1963 21:533−51)に記載の方法に沿って測定を行った。具体的には、所定箇所の6歯の舌側、唇側・頬側、近心側、遠心側の合計24歯面を目視により確認し、下記の評価基準及び評価方法で測定した。なお、上記所定箇所の6歯とは、上顎右側第1大臼歯、上顎左側中切歯、上顎左側第1小臼歯、下顎左側第1大臼歯、下顎右側中切歯、下顎右側第1小臼歯の6歯を指す。
0:臨床的に正常な歯肉。
1:軽度の炎症、歯肉のわずかな色調変化があるもので、プローブによる歯肉辺縁内縁部の擦過により出血が認められない。
2:中等度の炎症、歯肉に発赤を伴う浮腫と光沢があり、歯肉辺縁内縁部の擦過により出血が認められる。
3:高度の炎症、著しい発赤、浮腫があり、自然出血、潰瘍形成がある。
被験者らの刺激唾液(ガムを噛んで出る唾液)を5分間採取した後、歯科医師による問診と口腔診断を行った記録から、歯肉炎指数を算出した。歯肉炎指数は、下記の式により算出し、スクリーニング日(摂取開始日の1ヶ月前)及び摂取開始から5週間後に行った。
(歯肉炎指数)=(各歯の歯肉炎評価の合計)/(被検歯面数)
プラーク指数は、本発明の剤が歯周病予防効果及び歯周病治療効果を有するか否かの指標となるもので、非特許文献2(Comparative cleaning efficiency of manual and power brushing.Quigley H,Hein JN.,J Am Dent Ass 1962 65:26)に記載の方法に沿って測定を行った。具体的には、所定箇所の6歯の舌側、唇側・頬側、近心側、遠心側の合計24歯面を目視により確認し、下記の評価基準及び評価方法で測定した。なお、上記所定箇所の6歯とは、上顎右側第1大臼歯、上顎左側中切歯、上顎左側第1小臼歯、下顎左側第1大臼歯、下顎右側中切歯、下顎右側第1小臼歯の6歯を指す。
0:プラークの付着が認められない。
1:歯肉縁に沿ってプラークが点状に存在。
2:歯肉縁に沿ってプラークが線状に存在。
3:歯面の歯肉側3分の1以内にプラークが存在。
4:歯面の歯肉側3分の2以内にプラークが存在。
5:歯面の歯肉側3分の2以上にプラークが存在。
被験者の刺激唾液(ガムを噛んで出る唾液)を5分間採取した後、歯科医師による問診と口腔診断を行った記録から、プラーク指数を算出した。プラーク指数は、下記の式により算出し、スクリーニング日(摂取開始日の1ヶ月前)及び摂取開始から5週間後に行った。
(プラーク指数)=(各歯のプラーク評価の合計)/(被検歯面数)
1.う蝕菌比率
本発明の剤摂取群では、摂取開始日のう蝕菌比率の平均が0.5であったのに対し、4週間後のう蝕菌比率の平均は0.2と有意に低下した。一方、プラセボ摂取群では、摂取開始日のう蝕菌比率の平均が0.2であったのに対し、4週間後のう蝕菌比率の平均は0.2と有意な変化はなかった。
なお、本発明の剤摂取群とプラセボ摂取群の両群間の比較として、統計解析(両群間の比較はWilcoxon順位和検定、各群の摂取期間前後の比較はWilcoxon符号付順位検定)を行ったところ、摂取開始日のp値は0.218、4週間後のp値は0.973であった。
本発明の剤摂取群では、摂取開始日の歯肉炎指数の平均が0.13であったのに対し、5週間後の歯肉炎指数の平均は0.04と有意に低下した。一方、プラセボ摂取群では、摂取開始日の歯肉炎指数の平均が0.21であったのに対し、5週間後の歯肉炎指数の平均は0.505と有意な低下は見られなかった。
なお、本発明の剤摂取群とプラセボ摂取群の両群間の比較として、統計解析(両群間の比較はWilcoxon順位和検定、各群の摂取期間前後の比較はWilcoxon符号付順位検定)を行ったところ、スクリーニング日のp値は0.378であり、5週間後のp値は0.065であった。
本発明の剤摂取群では、摂取開始日のプラーク指数の平均が0.17であったのに対し、5週間後のプラーク指数の平均は0.17と有意な上昇は見られなかった。一方、プラセボ摂取群では、摂取開始日のプラーク指数の平均が0.17であったのに対し、5週間後のプラーク指数の平均は0.33と有意な高い値となった。
なお、本発明の剤摂取群とプラセボ摂取群の両群間の比較として、統計解析(両群間の比較はWilcoxon順位和検定、各群の摂取期間前後の比較はWilcoxon符号付順位検定)を行ったところ、スクリーニング日のp値は0.849であり、5週間後のp値は0.043であった。
また、歯肉炎指数の有意な低下とプラーク指数の悪化防止が、本発明の剤摂取群のみで認められプラセボ摂取群では認められなかったことから、本発明の剤は歯周病予防効果及び歯周病治療効果を有していることが分かった。
Claims (12)
- ビフィズス菌を含む抗う蝕剤。
- 前記ビフィズス菌がビフィドバクテリウム・ビフィダム(Bifidobacterium bifidum)である請求項1に記載の抗う蝕剤。
- 前記ビフィズス菌がビフィドバクテリウム・ビフィダムOLB6378菌株(Bifidobacterium bifidum OLB6378、受託番号:NITE BP−31)である請求項1又は2に記載の抗う蝕剤。
- 前記ビフィズス菌が加熱処理された菌体である、請求項1〜3のいずれか1項に記載の抗う蝕剤。
- 一日当たり108個以上の菌数の前記ビフィズス菌を4週間以上連続して適用される請求項1〜4のいずれか1項に記載の抗う蝕剤。
- 請求項1〜5のいずれか1項に記載の抗う蝕剤を含有する抗う蝕用組成物。
- さらに分散剤を含有する、請求項6に記載の抗う蝕用組成物。
- 前記分散剤がデキストリンである、請求項7に記載の抗う蝕用組成物。
- 請求項6〜8のいずれか1項に記載の抗う蝕用組成物と包材を含み、該抗う蝕用組成物が包材内に包装される、包装体。
- ビフィズス菌を含む歯周病予防剤又は歯周病治療剤。
- ビフィズス菌を含む抗う蝕剤、歯周病予防剤又は歯周病治療剤。
- 加熱処理されたビフィドバクテリウム・ビフィダムOLB6378菌株(Bifidobacterium bifidum OLB6378、受託番号:NITE BP−31)。
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