JP3665049B2 - Cell activator, antioxidant, and skin external preparation - Google Patents
Cell activator, antioxidant, and skin external preparation Download PDFInfo
- Publication number
- JP3665049B2 JP3665049B2 JP2002301489A JP2002301489A JP3665049B2 JP 3665049 B2 JP3665049 B2 JP 3665049B2 JP 2002301489 A JP2002301489 A JP 2002301489A JP 2002301489 A JP2002301489 A JP 2002301489A JP 3665049 B2 JP3665049 B2 JP 3665049B2
- Authority
- JP
- Japan
- Prior art keywords
- asclepias
- extract
- milkweed
- antioxidant
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
Description
【0001】
【発明の属する技術分野】
本発明は、細胞賦活剤、抗酸化剤、並びに係る細胞賦活剤及び抗酸化剤を含有する皮膚外用剤に関する。さらに詳しくは、ガガイモ科(Asclepiadaceae)のトウワタ属(Asclepias)植物抽出物を有効成分とする細胞賦活剤及び抗酸化剤、並びに係る細胞賦活剤及び抗酸化剤を含有し、老化症状の防止・改善に優れた効果を発揮する皮膚外用剤に関する。
【0002】
【従来の技術】
加齢などによる真皮線維芽細胞の機能低下は、コラーゲンやエラスチン等の真皮マトリックスの減少や変性を惹き起こし、シワや皮膚の弾性低下といった老化症状の重要な要因となっている。また、紫外線等の外来ストレスによる酸化傷害も、シワ,シミ,皮膚の弾性低下といった老化症状の原因となっている。これまでの皮膚外用剤の分野では、係る細胞の機能低下や酸化傷害による老化症状を防止・改善するために、様々な細胞賦活剤や抗酸化剤の検索及び配合検討がなされてきた。細胞賦活剤としては、ポンカンのエッセンス(特許文献1参照)、ツリガネニンジン属,クサギ及びそれらの抽出物(特許文献2参照)、有機溶媒によるクロレラ抽出物(特許文献3参照)が知られており、抗酸化剤としては、キク科へテロテカ属植物抽出物(特許文献4参照)、サルオガセ科サルオガセ属植物の抽出物(特許文献5参照)が知られている。
【0003】
なお、本発明に係るガガイモ科のトウワタ属植物の利用に関しては、アスクレピアス属(トウワタ属)の溶媒抽出物を消臭成分として配合してなる口腔用組成物が開示されている(特許文献6参照)。しかし、この文献公知発明には、ガガイモ科(Asclepiadaceae)のトウワタ属(Asclepias)植物抽出物の細胞賦活剤及び抗酸化剤としての利用や老化症状の防止・改善のために皮膚外用剤へ配合することに関する記載は全く認められなかった。
【0004】
【特許文献1】
特開2001−131045号公報
【特許文献2】
特開2000−178198号公報
【特許文献3】
特開平11−335293号公報
【特許文献4】
特開平11−180886号公報
【特許文献5】
特開平10−182413号公報
【特許文献6】
特開昭64−26512号公報
【0005】
【発明が解決しようとする課題】
従来用いられている細胞賦活剤及び抗酸化剤は、老化現象の一部の過程にのみ作用している場合が多く、本質的な老化改善効果としては不十分であると考えられた。また、皮膚外用剤の基剤中に配合した場合、有効な効果を得るにはかなりの高濃度を配合しなければならず、製剤に好ましくない色や臭いを付与してしまう場合があるなど、作用効果や安定性の面ですべてを満足できるものが少ないのが現状であった。このため、より優れた有効成分の開発が期待されており、本発明はこのような事情に鑑みてなされたものである。従って、本発明の目的は、優れた効果を有する細胞賦活剤及び優れた効果を有する抗酸化剤、並びに係る細胞賦活剤及び抗酸化剤を含有し、老化症状の防止・改善に優れた効果を発揮する皮膚外用剤を提供することにある。
【0006】
【課題を解決するための手段】
本発明者らは、皮膚の老化症状の防止・改善に優れた成分を見出すために、種々の物質について細胞賦活作用と抗酸化作用に関する検討を行った。その結果、トウワタ属(Asclepias)植物抽出物に優れた真皮線維芽細胞賦活作用と抗酸化作用を見出し、さらに検討を重ね、本発明を完成するに至った。
【0007】
【発明の実施の形態】
本発明の原料として用いられる植物は、ガガイモ科(Asclepiadaceae)のトウワタ属(Asclepias)の植物であればよい。トウワタ属(Asclepias)植物としては、トウワタ(Asclepias curassavica L.),オオトウワタ(Asclepias syriaca L.),ヤナギトウワタ(Asclepias tuberosa L.),アスクレピアス シラカ(Asclepias syraca),アスクレピアス スペキオサ(Asclepias speciosa)など約120種ほどが知られている。
【0008】
本発明に用いられる原料となる植物は、ガガイモ科(Asclepiadaceae)のトウワタ属(Asclepias)の植物であれば特に限定されないが、入手が比較的容易などの理由から原料として適当なものとしては、トウワタ(Asclepias curassavica L.),オオトウワタ(Asclepias syriaca L.),ヤナギトウワタ(Asclepias tuberosa L.)などが挙げられる。
【0009】
これらトウワタ属(Asclepias)植物を使用する際は、抽出物を用いるのが一般的である。抽出には、トウワタ属(Asclepias)植物の根,葉,花,茎,幹皮などのいずれの部位を用いても構わないが、簡便に利用するには全草を用いるとよい。抽出の際は、生のまま用いてもよいが、抽出効率を考えると、細切,乾燥,粉砕等の処理を行った後に抽出を行うことが好ましい。抽出は、抽出溶媒に浸漬するか、超臨界流体や亜臨界流体を用いた抽出方法でも行うことができる。抽出効率を上げるため、撹拌や抽出溶媒中でホモジナイズしてもよい。抽出温度は、特に限定はされないが、5℃程度から抽出溶媒の沸点以下の温度とするのが好ましい。抽出時間は抽出溶媒の種類や抽出温度によっても異なるが、1時間〜14日間程度とするのが好ましい。
【0010】
抽出溶媒としては、水の他、メタノール,エタノール,プロパノール,イソプロパノール等の低級アルコール、1,3−ブチレングリコール,プロピレングリコール,ジプロピレングリコール,グリセリン等の多価アルコール、エチルエーテル,プロピルエーテル等のエーテル類、酢酸ブチル,酢酸エチル等のエステル類、アセトン,エチルメチルケトン等のケトン類などの溶媒を用いることができ、これらより1種又は2種以上を選択して用いる。また、生理食塩水,リン酸緩衝液,リン酸緩衝生理食塩水等を用いてもよい。さらに、水や二酸化炭素,エチレン,プロピレン,エタノール,メタノール,アンモニアなどの1種又は2種以上の超臨界流体や亜臨界流体を用いてもよい。
【0011】
トウワタ属(Asclepias)植物の上記溶媒による抽出物は、そのままでも使用することができるが、濃縮,乾固した物を水や極性溶媒に再度溶解したり、或いはこれらの生理作用を損なわない範囲で脱色,脱臭,脱塩等の精製処理を行ったり、カラムクロマトグラフィー等による分画処理を行った後に用いてもよい。トウワタ属(Asclepias)植物の前記抽出物やその処理物及び分画物は、各処理及び分画後に凍結乾燥し、用時に溶媒に溶解して用いることもできる。また、リポソーム等のベシクルやマイクロカプセル等に内包させて用いることもできる。
【0012】
本発明における細胞賦活剤及び抗酸化剤は、上述のトウワタ属(Asclepias)植物抽出物を有効成分とする。また、係るトウワタ属(Asclepias)植物抽出物を細胞賦活剤及び抗酸化剤として皮膚外用剤に配合することにより、老化症状の予防・改善に優れた効果を発揮する。
【0013】
本発明におけるトウワタ属(Asclepias)植物抽出物の配合量は、皮膚外用剤の種類や目的等によって調整することができるが、皮膚外用剤の全量に対して、0.0001〜10.0重量%が好ましく、より好ましくは、0.001〜5.0重量%である。
【0014】
本発明に係る皮膚外用剤は、ローション,乳液,ゲル,クリーム,軟膏剤,粉末,顆粒等、種々の剤型で提供することができる。
【0015】
なお、本発明に係る皮膚外用剤には、トウワタ属(Asclepias)植物抽出物の他に、通常医薬品,医薬部外品,皮膚化粧料,毛髪用化粧料及び洗浄料に配合される、油性成分,保湿剤,粉体,色素,乳化剤,可溶化剤,洗浄剤,紫外線吸収剤,増粘剤,薬剤,香料,樹脂,防菌防黴剤,アルコール類等を適宜配合することができる。また、本発明の効果を損なわない範囲において、他の細胞賦活剤,抗酸化剤,植物抽出物との併用も可能である。
【0016】
【実施例】
さらに実施例により、本発明の特徴について詳細に説明する。まず、本発明のトウワタ属(Asclepias)植物抽出物の調製方法について示す。
【0017】
[調製方法1]
トウワタ属(Asclepias)植物の全草の乾燥粉砕物1kgに50重量%エタノール水溶液を10リットル加え、室温で7日間浸漬した。抽出液をろ過して回収し、溶媒を除去した後、トウワタ属(Asclepias)植物抽出物を得た。
【0018】
[調製方法2]
トウワタ属(Asclepias)植物の全草の乾燥粉砕物1kgに水を9リットル加え、90℃にて6時間還流して抽出した。抽出液をろ過して回収し、溶媒を除去した後、トウワタ属(Asclepias)植物抽出物を得た。
【0019】
[調製方法3]
トウワタ属(Asclepias)植物の全草の乾燥粉砕物1kgにメタノールを9リットル加え、室温で7日間浸漬した。抽出液をろ過して回収し、溶媒を除去した後、トウワタ属(Asclepias)植物抽出物を得た。
【0020】
[調製方法4]
超臨界抽出装置にトウワタ属(Asclepias)植物の全草を投入し、40℃において15MPaの気圧下で二酸化炭素の超臨界流体を用いて抽出した。抽出物を回収し、トウワタ属(Asclepias)植物抽出物を得た。
【0021】
次に、トウワタ属(Asclepias)植物抽出物の真皮線維芽細胞賦活作用を示す。試料には、トウワタ(Asclepias curassavicaL.)より調製方法1を用いて抽出したトウワタ抽出物を用いた。
【0022】
評価は、以下の手順で行った。正常ヒト真皮線維芽細胞を1ウェル当たり2.0×104個となるように96穴マイクロプレートに播種した。播種培地には、ダルベッコ改変イーグル培地(DMEM)に1%のウシ胎児血清を添加したものを用いた。24時間培養後、任意の濃度の試料を添加した試験培地に交換し、さらに48時間培養した。次いで3−(4,5−ジメチル−2−チアゾリル)−2,5−ジフェニルテトラゾリウムブロミド(MTT)を400μg/mL含有する培地に交換して2時間培養し、テトラゾリウム環の開環により生じるフォルマザンを2−プロパノールにて抽出し、マイクロプレートリーダーにて550nmの吸光度を測定した。同時に濁度として650nmにおける吸光度を測定し、両測定値の差により細胞賦活作用を評価した。また、測定法の妥当性を確認するために、試料を添加した培地の代わりに、ダルベッコ改変イーグル培地(DMEM)に5%のウシ胎児血清を添加したものを陽性コントロールとし、測定を行った。評価結果を、試料無添加のブランクにおける細胞賦活作用を100とした相対値にて表1に示す。
【0023】
【表1】
【0024】
表1より明らかなように、トウワタ抽出物を添加した培地では、有意な真皮線維芽細胞賦活作用が認められた。特に、トウワタ抽出物を8.0μg/mL添加した場合に、ブランクと比較して、危険率1%未満で有意な真皮線維芽細胞賦活作用が認められた。このことから、トウワタ抽出物は、優れた真皮線維芽細胞賦活作用を有することが明らかとなった。
【0025】
次に、トウワタ属(Asclepias)植物抽出物の抗酸化作用について示す。試料には、トウワタ(Asclepias curassavica L.)より調製方法1を用いて抽出したトウワタ抽出物を用いた。
【0026】
評価は、以下の手順で行った。50重量%エタノール水溶液にて1mg/mLに希釈したトウワタ抽出物溶液を96穴マイクロプレートに100μL添加した。次に、0.2mMの濃度になるようにエタノールにて調製した1,1−ジフェニル−2−ピクリルヒドラジル(DPPH)溶液を96穴マイクロプレートに100μL添加した。攪拌しながら暗所に放置し、24時間後に516nmの吸光度を測定した。試料が無添加のブランクの吸光度を(A)、試料を添加したときの吸光度を(B)としたとき、式(1)の値をラジカル消去率とした。
式(1) {1−(B)/(A)}×100(%)
トウワタ抽出物の実験結果を表2に示す。
【0027】
【表2】
【0028】
表2より明らかなように、トウワタ抽出物は優れた抗酸化作用を有することが分かった。
【0029】
続いて、本発明に係るトウワタ属(Asclepias)植物抽出物を配合した皮膚外用剤の処方を示す。実施例1〜5の処方には、表3に示すトウワタ属(Asclepias)植物抽出物を配合し、実施例6〜18の各処方には、それぞれの処方に記載のトウワタ属(Asclepias)植物を配合した。
【0030】
製法:(1)〜(6)の油相成分を80℃にて加熱溶解する。一方(7)〜(10)の水相成分を80℃にて加熱溶解する。これに前記油相成分を攪拌しながら加え、ホモジナイザーにより均一に乳化する。乳化終了後、冷却を開始し、(11)と(12)を順次加え、均一に混合する。
【0031】
【表3】
【0032】
[実施例6]化粧水
(1)エタノール 15.0(重量%)
(2)ポリオキシエチレン(40E.O.)硬化ヒマシ油 0.3
(3)香料 0.1
(4)精製水 81.36
(5)クエン酸 0.02
(6)クエン酸ナトリウム 0.1
(7)グリセリン 3.0
(8)ヒドロキシエチルセルロース 0.1
(9)トウワタ抽出物[調製方法3] 0.02
製法:(1)に(2)及び(3)を溶解する。溶解後、(4)〜(8)を順次添加した後、十分に攪拌し、(9)を加え、均一に混合する。
【0033】
[実施例7]クリーム
(1)スクワラン 10.0(重量%)
(2)ステアリン酸 2.0
(3)水素添加パーム核油 0.5
(4)水素添加大豆リン脂質 0.1
(5)セタノール 3.6
(6)親油型モノステアリン酸グリセリン 2.0
(7)グリセリン 10.0
(8)パラオキシ安息香酸メチル 0.1
(9)アルギニン(20重量%水溶液) 15.0
(10)精製水 40.7
(11)カルボキシビニルポリマー(1重量%水溶液) 15.0
(12)トウワタ抽出物[調製方法1] 1.0
製法:(1)〜(6)の油相成分を80℃にて加熱溶解する。一方(7)〜(10)の水相成分を80℃にて加熱溶解する。これに前記油相成分を攪拌しながら加え、ホモジナイザーにより均一に乳化する。乳化終了後、(11)を加え、冷却を開始し、40℃にて(12)を加え、均一に混合する。
【0034】
製法:(1)〜(6)の水相成分を混合し、75℃にて加熱溶解する。一方、(7)〜(14)の油相成分を混合し、75℃にて加熱溶解する。次いで、上記水相成分に油相成分を添加して予備乳化を行った後、ホモミキサーにて均一に乳化する。乳化終了後に冷却を開始し、50℃にて(15)を加える。さらに40℃まで冷却し、(16)を加え、均一に混合する。
【0035】
[実施例9]水性ジェル
(1)カルボキシビニルポリマー 0.5(重量%)
(2)精製水 86.4
(3)水酸化ナトリウム(10重量%水溶液) 0.5
(4)エタノール 10.0
(5)パラオキシ安息香酸メチル 0.1
(6)香料 0.1
(7)トウワタ抽出物[調製方法4] 0.1
(8)ポリオキシエチレン(60E.O.)硬化ヒマシ油 0.1
(9)グリセリン 2.0
(10)ヤナギトウワタ抽出物[調製方法3] 0.1
(11)オオトウワタ抽出物[調製方法2] 0.1
製法:(1)を(2)に加え、均一に攪拌した後、(3)を加える。均一に攪拌した後,(4)に予め溶解した(5)を加える。均一に攪拌した後、予め混合しておいた(6)〜(8)を加える。均一に攪拌した後、(9)〜(11)を順次加え、均一に混合する。
【0036】
[実施例10]クレンジング料
(1)スクワラン 81.9(重量%)
(2)イソステアリン酸ポリオキシエチレングリセリル 15.0
(3)精製水 3.0
(4)トウワタ抽出物[調製方法4] 0.05
(5)ヤナギトウワタ抽出物[調製方法4] 0.05
製法:(1)と(2)を均一に溶解する。これに、(3)〜(5)を順次加え、均一に混合する。
【0037】
[実施例11]洗顔フォーム
(1)ステアリン酸 16.0(重量%)
(2)ミリスチン酸 16.0
(3)親油型モノステアリン酸グリセリン 2.0
(4)グリセリン 20.0
(5)水酸化ナトリウム 7.5
(6)ヤシ油脂肪酸アミドプロピルベタイン 1.0
(7)精製水 36.5
(8)トウワタ抽出物[調製方法3] 0.5
(9)オオトウワタ抽出物[調製方法2] 0.5
製法:(1)〜(4)の油相成分を80℃にて加熱溶解する。一方(5)〜(7)の水相成分を80℃にて加熱溶解し、油相成分と均一に混合撹拌する。冷却を開始し、40℃にて(8)と(9)を加え、均一に混合する。
【0038】
[実施例12]メイクアップベースクリーム
(1)スクワラン 10.0(重量%)
(2)セタノール 2.0
(3)グリセリントリ−2−エチルヘキサン酸エステル 2.5
(4)親油型モノステアリン酸グリセリル 1.0
(5)プロピレングリコール 11.0
(6)ショ糖脂肪酸エステル 1.3
(7)精製水 70.4
(8)酸化チタン 1.0
(9)ベンガラ 0.1
(10)黄酸化鉄 0.4
(11)香料 0.1
(12)トウワタ抽出物[調製方法2] 0.1
(13)ヤナギトウワタ抽出物[調製方法1] 0.1
製法:(1)〜(4)の油相成分を混合し、75℃にて加熱溶解する。一方、(5)〜(7)の水相成分を混合し、75℃にて加熱溶解し、これに(8)〜(10)の顔料を加え、ホモミキサーにて均一に分散させる。この水相成分に前記油相成分を加え、ホモミキサーにて乳化する。乳化終了後に冷却を開始し、40℃にて(11)〜(13)の成分を加え、均一に混合する。
【0039】
製法:(1)〜(6)の油相成分を混合し、75℃にて加熱溶解する。一方、(7)〜(10)の水相成分を混合し、75℃にて加熱溶解し、これに(11)〜(15)の顔料を加え、ホモミキサーにて均一に分散する。油相成分を加え、乳化を行う。乳化終了後に冷却を開始し、40℃にて(16)〜(18)の成分を順次加え、均一に混合する。
【0040】
[実施例14]油中水型エモリエントクリーム
(1)流動パラフィン 30.0(重量%)
(2)マイクロクリスタリンワックス 2.0
(3)ワセリン 5.0
(4)ジグリセリンオレイン酸エステル 5.0
(5)塩化ナトリウム 1.3
(6)塩化カリウム 0.1
(7)プロピレングリコール 3.0
(8)1,3−ブチレングリコール 5.0
(9)パラオキシ安息香酸メチル 0.1
(10)トウワタ抽出物[調製方法1] 0.5
(11)精製水 45.9
(12)香料 0.1
(13)オオトウワタ抽出物[調製方法4] 2.0
製法:(5)と(6)を(11)の一部に溶解して50℃とし、50℃に加熱した(4)に撹拌しながら徐々に加える。これを混合した後、70℃にて加熱溶解した(1)〜(3)に均一に分散する。これに(7)〜(10)を(11)の残部に70℃にて加熱溶解したものを撹拌しながら加え、ホモミキサーにて乳化する。乳化終了後に冷却を開始し、40℃にて(12)と(13)を加え、均一に混合する。
【0041】
[実施例15]ヘアートニック
(1)エタノール 50.0(重量%)
(2)精製水 49.898
(3)オオトウワタ抽出物[調製方法3] 0.001
(4)トウワタ抽出物[調製方法4] 0.001
(5)香料 0.1
製法:(1)〜(5)の成分を混合,均一化する。
【0042】
[実施例16]パック
(1)精製水 68.9(重量%)
(2)ポリビニルアルコール 12.0
(3)エタノール 10.0
(4)グリセリン 5.0
(5)ポリエチレングリコール(平均分子量1000) 2.0
(6)ヤナギトウワタ抽出物[調製方法1] 1.0
(7)トウワタ抽出物[調製方法2] 1.0
(8)香料 0.1
製法:(2)と(3)を混合し、80℃に加温した後、80℃に加温した(1)に溶解する。均一に溶解した後、(4)〜(5)を加え、攪拌しながら冷却を開始する。40℃まで冷却し、(6)〜(8)を加え、均一に混合する。
【0043】
製法:(1)〜(6)の油相成分を混合し、75℃にて加熱溶解後する。一方、(7)〜(10)の水相成分を75℃にて加熱溶解し、前記油相成分を加え、ホモミキサーにて乳化する。乳化終了後に冷却を開始し、40℃にて(11)〜(13)の成分を加え、均一に混合する。
【0044】
[実施例18]入浴剤
(1)香料 0.3(重量%)
(2)トウワタ抽出物[調製方法1] 0.5
(3)オオトウワタ抽出物[調製方法3] 0.5
(4)炭酸水素ナトリウム 50.0
(5)硫酸ナトリウム 48.7
製法:(1)〜(5)を均一に混合する。
【0045】
本発明の実施例1〜5について使用試験を行い、シワ,タルミ,肌のハリの改善効果を評価した。その際、実施例1において、配合したトウワタ抽出物を精製水に代替し、比較例1として同時に使用試験を行った。
【0046】
各試料について、シワ,タルミ,肌のハリの低下といった症状が顕著に認められる50〜60才代の男女パネラー各20名を一群とし、ブラインドにて1カ月間使用させ、使用前後の皮膚状態の変化を観察して評価した。皮膚症状の指標として、シワ,タルミ,肌のハリについて、「改善」,「やや改善」,「変化なし」の三段階で評価し、表4に各評価を得たパネラー数にて示した。
【0047】
【表4】
【0048】
表4より、シワ,タルミ,肌のハリについて、トウワタ属(Asclepias)植物抽出物を含有しない比較例使用群においては、半数以上のパネラーに改善が認められなかったが、トウワタ属(Asclepias)植物抽出物を配合した実施例使用群においては、6割以上のパネラーに明確な改善が認められた。
【0049】
以上のように、本発明の実施例においては、従来の比較例よりも、シワ,タルミ,肌のハリといった老化症状の改善に優れた効果を有していた。
【0050】
【発明の効果】
以上詳述したように、本発明により、優れた効果を有する細胞賦活剤及び抗酸化剤を得ることが出来た。また、係る細胞賦活剤及び抗酸化剤を有効成分として配合することにより、皮膚の老化症状の防止・改善に優れた効果を有する皮膚外用剤を得ることが出来た。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a cell activator, an antioxidant, and a skin external preparation containing such a cell activator and an antioxidant. More particularly, contains Asclepias (Asclepias) cell activator and an antioxidant as an active ingredient a plant extract, cell activators according to the arrangement and antioxidants Asclepiadaceae (Asclepiadaceae), prevention and improvement of senile symptoms The present invention relates to a skin external preparation that exhibits an excellent effect on skin.
[0002]
[Prior art]
A decrease in the function of dermal fibroblasts due to aging causes a decrease or degeneration of the dermal matrix such as collagen and elastin, and is an important factor of aging symptoms such as wrinkles and a decrease in skin elasticity. In addition, oxidative damage caused by extraneous stress such as ultraviolet rays also causes aging symptoms such as wrinkles, spots, and a decrease in skin elasticity. Until now, in the field of topical skin preparations, various cell activators and antioxidants have been searched for and investigated in order to prevent and improve the aging symptoms caused by such functional deterioration of cells and oxidative damage. As cell activators, the essence of Ponkan (refer to Patent Document 1), genus Genus, Wedge and their extracts (refer to Patent Document 2), chlorella extract using an organic solvent (refer to Patent Document 3), As antioxidants, Asteraceae heterotheca plant extracts (see Patent Document 4) and Salogaceae Salogase plant extracts (see Patent Document 5) are known.
[0003]
In addition, regarding the utilization of the plant of the genus Calyceae which concerns on this invention, the composition for oral cavity formed by mix | blending the solvent extract of the genus Asclepias (genus Towata) as a deodorizing component is disclosed (patent document 6). reference). However, this literature known invention, formulated into Asclepias (Asclepias) skin external agent for the prevention and improvement of use and aging symptoms as a cell activator and antioxidant plant extracts of Asclepiadaceae (Asclepiadaceae) There was no statement regarding this.
[0004]
[Patent Document 1]
JP 2001-131045 A [Patent Document 2]
JP 2000-178198 A [Patent Document 3]
Japanese Patent Laid-Open No. 11-335293 [Patent Document 4]
JP-A-11-180886 [Patent Document 5]
JP-A-10-182413 [Patent Document 6]
JP-A 64-26512
[Problems to be solved by the invention]
Conventionally used cell activators and antioxidants often act only in part of the aging phenomenon, and are considered to be insufficient as an essential aging improvement effect. In addition, when blended into the base of the external preparation for skin, to obtain an effective effect, it must be blended in a fairly high concentration, and may give an unfavorable color and odor to the formulation, etc. At present, there are few things that can satisfy all of the effects and stability. For this reason, development of a more effective active ingredient is anticipated and this invention is made | formed in view of such a situation. Therefore, the object of the present invention is to contain a cell activator having an excellent effect and an antioxidant having an excellent effect, and such a cell activator and an antioxidant, and has an excellent effect in preventing and improving aging symptoms. It is in providing the skin external preparation which exhibits.
[0006]
[Means for Solving the Problems]
In order to find a component excellent in prevention and improvement of skin aging symptoms, the present inventors have studied cell activation and antioxidant effects of various substances. As a result, an excellent dermal fibroblast activation action and antioxidant action were found in the Asclepias plant extract, and further studies were made to complete the present invention.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
Plants used as a raw material of the present invention may be any plant of Asclepias of Asclepiadaceae (Asclepiadaceae) (Asclepias). The Asclepias (Asclepias) plants, milkweed (Asclepias curassavica L.), Ootouwata (Asclepias syriaca L.), pleurisy (Asclepias tuberosa L.), Asclepias Shiraka (Asclepias syraca), Asclepias Supekiosa (Asclepias speciosa), etc. About 120 species are known.
[0008]
Plants as a raw material used in the present invention is not particularly limited as long as it is a plant of Asclepias of Asclepiadaceae (Asclepiadaceae) (Asclepias), as being suitable as a raw material for reasons such as availability is relatively easy, milkweed ( Asclepias curassavica L.), Japanese cotton ( Asclepias syriaca L.), Willow milkweed ( Asclepias tuberosa L.) and the like.
[0009]
When using these Asclepias plants, it is common to use an extract. For extraction, any part of the roots, leaves, flowers, stems, stem bark, etc. of Asclepias plants may be used, but whole plants may be used for convenient use. In the extraction, it may be used as it is, but considering the extraction efficiency, it is preferable to perform the extraction after performing processing such as shredding, drying, and pulverization. The extraction can be performed by immersing in an extraction solvent or by an extraction method using a supercritical fluid or a subcritical fluid. In order to increase the extraction efficiency, the mixture may be homogenized in stirring or an extraction solvent. Although extraction temperature is not specifically limited, It is preferable to set it as the temperature below about the boiling point of an extraction solvent from about 5 degreeC. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but is preferably about 1 hour to 14 days.
[0010]
Extraction solvents include water, lower alcohols such as methanol, ethanol, propanol, and isopropanol, polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin, and ethers such as ethyl ether and propyl ether. , Solvents such as esters such as butyl acetate and ethyl acetate, and ketones such as acetone and ethyl methyl ketone can be used, and one or more of these are selected and used. Further, physiological saline, phosphate buffer, phosphate buffered saline, or the like may be used. Furthermore, you may use 1 type, or 2 or more types of supercritical fluids and subcritical fluids, such as water, a carbon dioxide, ethylene, propylene, ethanol, methanol, ammonia.
[0011]
An extract of Asclepias plants using the above solvent can be used as it is, but the concentrated and dried product can be dissolved again in water or a polar solvent, or the physiological action thereof is not impaired. You may use, after performing purification processes, such as decoloring, deodorizing, and desalting, or performing the fractionation process by column chromatography etc. The above-mentioned extract of the genus Asclepias , its treated product and fractions can be freeze-dried after each treatment and fractionation and dissolved in a solvent before use. It can also be used by encapsulating in vesicles such as liposomes or microcapsules.
[0012]
The cell activator and antioxidant in the present invention contain the above-mentioned Asclepias plant extract as an active ingredient. Moreover, the effect which was excellent in the prevention and improvement of an aging symptom is shown by mix | blending the plant extract ( Asclepias ) which concerns with a skin external preparation as a cell activator and an antioxidant.
[0013]
The blending amount of the plant extract of the genus Asclepias in the present invention can be adjusted according to the type and purpose of the external preparation for skin, but is 0.0001 to 10.0% by weight based on the total amount of the external preparation for skin. Is more preferable, and 0.001 to 5.0% by weight is more preferable.
[0014]
The external preparation for skin according to the present invention can be provided in various dosage forms such as lotion, emulsion, gel, cream, ointment, powder, granule and the like.
[0015]
It should be noted that the skin external preparation according to the present invention includes oily ingredients that are usually blended in pharmaceuticals, quasi-drugs, skin cosmetics, hair cosmetics, and cleansing agents in addition to plant extracts of the genus Asclepias. , Moisturizers, powders, pigments, emulsifiers, solubilizers, detergents, UV absorbers, thickeners, drugs, fragrances, resins, antibacterial / antifungal agents, alcohols, and the like can be appropriately blended. Moreover, in the range which does not impair the effect of this invention, combined use with another cell activator, an antioxidant, and a plant extract is also possible.
[0016]
【Example】
Further, the features of the present invention will be described in detail by way of examples. First, the method for preparing the Asclepias plant extract of the present invention will be described.
[0017]
[Preparation Method 1]
10 kg of 50% by weight aqueous ethanol solution was added to 1 kg of a dry pulverized whole plant of Asclepias plants, and immersed for 7 days at room temperature. The extract was collected by filtration, and after removing the solvent, an Asclepias plant extract was obtained.
[0018]
[Preparation Method 2]
Nine liters of water was added to 1 kg of a dry pulverized whole plant of Asclepias plants, and the mixture was extracted by refluxing at 90 ° C. for 6 hours. The extract was collected by filtration, and after removing the solvent, an Asclepias plant extract was obtained.
[0019]
[Preparation Method 3]
Nine liters of methanol was added to 1 kg of a dry pulverized whole plant of Asclepias plants and immersed for 7 days at room temperature. The extract was collected by filtration, and after removing the solvent, an Asclepias plant extract was obtained.
[0020]
[Preparation Method 4]
The whole plant of Asclepias plant was put into a supercritical extraction apparatus, and extracted using a supercritical fluid of carbon dioxide at 40 ° C. under a pressure of 15 MPa. The extract was collected to obtain an Asclepias plant extract.
[0021]
Next, the dermal fibroblast activation action of the Asclepias plant extract is shown. As a sample, milkweed extract extracted from milkweed ( Asclepias curassavica L.) using preparation method 1 was used.
[0022]
The evaluation was performed according to the following procedure. Normal human dermal fibroblasts were seeded in a 96-well microplate at 2.0 × 10 4 cells per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 1% fetal bovine serum. After culturing for 24 hours, the culture medium was replaced with a test medium to which a sample having an arbitrary concentration was added, and further cultured for 48 hours. Next, the medium containing 400 μg / mL of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyltetrazolium bromide (MTT) was exchanged and cultured for 2 hours. Formazan produced by the opening of the tetrazolium ring was removed. Extraction was performed with 2-propanol, and absorbance at 550 nm was measured with a microplate reader. At the same time, the absorbance at 650 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values. Further, in order to confirm the validity of the measurement method, instead of the medium to which the sample was added, measurement was performed using Dulbecco's modified Eagle medium (DMEM) with 5% fetal bovine serum as a positive control. The evaluation results are shown in Table 1 as relative values with the cell activation effect in the blank with no sample as 100.
[0023]
[Table 1]
[0024]
As is clear from Table 1, a significant dermal fibroblast activation effect was observed in the medium supplemented with milkweed extract. In particular, when Towata extract was added at 8.0 μg / mL, a significant dermal fibroblast activation effect was observed at a risk rate of less than 1% compared to the blank. From this, it was revealed that milkweed extract has an excellent dermal fibroblast activation effect.
[0025]
Next, the antioxidant effect of the Asclepias plant extract is shown. As a sample, milkweed extract extracted from milkweed ( Asclepias curassavica L.) using preparation method 1 was used.
[0026]
The evaluation was performed according to the following procedure. 100 μL of milkweed extract solution diluted to 1 mg / mL with 50 wt% aqueous ethanol was added to a 96-well microplate. Next, 100 μL of a 1,1-diphenyl-2-picrylhydrazyl (DPPH) solution prepared with ethanol to a concentration of 0.2 mM was added to a 96-well microplate. The mixture was left in the dark with stirring, and the absorbance at 516 nm was measured after 24 hours. When the absorbance of the blank with no sample added was (A) and the absorbance when the sample was added was (B), the value of formula (1) was defined as the radical elimination rate.
Formula (1) {1- (B) / (A)} × 100 (%)
The experimental results of milkweed extract are shown in Table 2.
[0027]
[Table 2]
[0028]
As is apparent from Table 2, milkweed extract was found to have an excellent antioxidant effect.
[0029]
Then, the prescription of the skin external preparation which mix | blended the Asperias plant extract based on this invention is shown. The formulas of Examples 1 to 5 are blended with the plant extracts of Asclepias shown in Table 3, and the formulas of Examples 6 to 18 are the plants of the genus Asclepias described in the respective formulas. Blended.
[0030]
Production method: The oil phase components (1) to (6) are heated and dissolved at 80 ° C. On the other hand, the aqueous phase components (7) to (10) are dissolved by heating at 80 ° C. The oil phase component is added to this while stirring and uniformly emulsified with a homogenizer. After emulsification, start cooling and add (11) and (12) sequentially and mix uniformly.
[0031]
[Table 3]
[0032]
[Example 6] Lotion (1) Ethanol 15.0 (% by weight)
(2) Polyoxyethylene (40E.O.) hydrogenated castor oil 0.3
(3) Fragrance 0.1
(4) Purified water 81.36
(5) Citric acid 0.02
(6) Sodium citrate 0.1
(7) Glycerin 3.0
(8) Hydroxyethyl cellulose 0.1
(9) Milkweed extract [Preparation method 3] 0.02
Production method: (2) and (3) are dissolved in (1). After dissolution, (4) to (8) are sequentially added, and then sufficiently stirred, (9) is added and mixed uniformly.
[0033]
[Example 7] Cream (1) Squalane 10.0 (% by weight)
(2) Stearic acid 2.0
(3) Hydrogenated palm kernel oil 0.5
(4) Hydrogenated soybean phospholipid 0.1
(5) Cetanol 3.6
(6) Lipophilic glyceryl monostearate 2.0
(7) Glycerin 10.0
(8) Methyl paraoxybenzoate 0.1
(9) Arginine (20% by weight aqueous solution) 15.0
(10) Purified water 40.7
(11) Carboxyvinyl polymer (1% by weight aqueous solution) 15.0
(12) Milkweed extract [Preparation method 1] 1.0
Production method: The oil phase components (1) to (6) are heated and dissolved at 80 ° C. On the other hand, the aqueous phase components (7) to (10) are dissolved by heating at 80 ° C. The oil phase component is added to this while stirring and uniformly emulsified with a homogenizer. After the emulsification is completed, add (11), start cooling, add (12) at 40 ° C., and mix uniformly.
[0034]
Production method: The aqueous phase components (1) to (6) are mixed and dissolved by heating at 75 ° C. On the other hand, the oil phase components (7) to (14) are mixed and dissolved by heating at 75 ° C. Next, the oil phase component is added to the aqueous phase component and preliminary emulsification is performed, followed by uniform emulsification with a homomixer. Cooling is started after completion of emulsification, and (15) is added at 50 ° C. Cool further to 40 ° C, add (16) and mix evenly.
[0035]
[Example 9] Aqueous gel (1) Carboxyvinyl polymer 0.5 (% by weight)
(2) Purified water 86.4
(3) Sodium hydroxide (10% by weight aqueous solution) 0.5
(4) Ethanol 10.0
(5) Methyl paraoxybenzoate 0.1
(6) Fragrance 0.1
(7) Milkweed extract [Preparation method 4] 0.1
(8) Polyoxyethylene (60E.O.) hydrogenated castor oil 0.1
(9) Glycerin 2.0
(10) Willow milkweed extract [Preparation method 3] 0.1
(11) Milkweed extract [Preparation method 2] 0.1
Manufacturing method: (1) is added to (2), and after stirring uniformly, (3) is added. After stirring uniformly, add (5) previously dissolved in (4). After stirring uniformly, the previously mixed (6) to (8) are added. After stirring uniformly, (9) to (11) are sequentially added and mixed uniformly.
[0036]
[Example 10] Cleansing fee (1) Squalane 81.9 (% by weight)
(2) Polyoxyethylene glyceryl isostearate 15.0
(3) Purified water 3.0
(4) Milkweed extract [Preparation method 4] 0.05
(5) Willow milkweed extract [Preparation Method 4] 0.05
Manufacturing method: (1) and (2) are uniformly dissolved. To this, (3) to (5) are sequentially added and mixed uniformly.
[0037]
[Example 11] Face washing foam (1) Stearic acid 16.0 (wt%)
(2) Myristic acid 16.0
(3) Lipophilic glyceryl monostearate 2.0
(4) Glycerin 20.0
(5) Sodium hydroxide 7.5
(6) Palm oil fatty acid amidopropyl betaine 1.0
(7) Purified water 36.5
(8) Milkweed extract [Preparation method 3] 0.5
(9) Milkweed extract [Preparation method 2] 0.5
Production method: The oil phase components (1) to (4) are heated and dissolved at 80 ° C. On the other hand, the aqueous phase components (5) to (7) are heated and dissolved at 80 ° C., and mixed and stirred uniformly with the oil phase components. Cooling is started, and (8) and (9) are added at 40 ° C. and mixed uniformly.
[0038]
[Example 12] Make-up base cream (1) Squalane 10.0 (% by weight)
(2) Cetanol 2.0
(3) Glycerin tri-2-ethylhexanoate 2.5
(4) Lipophilic glyceryl monostearate 1.0
(5) Propylene glycol 11.0
(6) Sucrose fatty acid ester 1.3
(7) Purified water 70.4
(8) Titanium oxide 1.0
(9) Bengala 0.1
(10) Yellow iron oxide 0.4
(11) Fragrance 0.1
(12) Milkweed extract [Preparation method 2] 0.1
(13) Willow milkweed extract [Preparation method 1] 0.1
Production method: The oil phase components (1) to (4) are mixed and dissolved by heating at 75 ° C. On the other hand, the aqueous phase components (5) to (7) are mixed and dissolved by heating at 75 ° C., and the pigments (8) to (10) are added thereto and dispersed uniformly with a homomixer. The oil phase component is added to the water phase component and emulsified with a homomixer. Cooling is started after the emulsification is completed, and the components (11) to (13) are added at 40 ° C. and mixed uniformly.
[0039]
Production method: The oil phase components (1) to (6) are mixed and dissolved by heating at 75 ° C. On the other hand, the aqueous phase components (7) to (10) are mixed and dissolved by heating at 75 ° C., and the pigments (11) to (15) are added thereto and uniformly dispersed with a homomixer. Add oil phase ingredients and emulsify. Cooling is started after the emulsification is completed, and the components (16) to (18) are sequentially added at 40 ° C. and mixed uniformly.
[0040]
[Example 14] Water-in-oil emollient cream (1) Liquid paraffin 30.0 (wt%)
(2) Microcrystalline wax 2.0
(3) Vaseline 5.0
(4) Diglycerin oleate 5.0
(5) Sodium chloride 1.3
(6) Potassium chloride 0.1
(7) Propylene glycol 3.0
(8) 1,3-butylene glycol 5.0
(9) Methyl paraoxybenzoate 0.1
(10) Milkweed extract [Preparation method 1] 0.5
(11) Purified water 45.9
(12) Fragrance 0.1
(13) Milkweed extract [Preparation method 4] 2.0
Production method: Dissolve (5) and (6) in a part of (11) to 50 ° C., and gradually add to (4) heated to 50 ° C. with stirring. After mixing this, it disperse | distributes uniformly to (1)-(3) heated and melt | dissolved at 70 degreeC. (7) to (10) are added to the remainder of (11) heated and dissolved at 70 ° C. while stirring and emulsified with a homomixer. Cooling is started after the emulsification, and (12) and (13) are added at 40 ° C. and mixed uniformly.
[0041]
[Example 15] Hair artic (1) Ethanol 50.0 (wt%)
(2) Purified water 49.898
(3) Milkweed extract [Preparation method 3] 0.001
(4) Milkweed extract [Preparation method 4] 0.001
(5) Fragrance 0.1
Production method: Components (1) to (5) are mixed and homogenized.
[0042]
[Example 16] Pack (1) 68.9 (% by weight) purified water
(2) Polyvinyl alcohol 12.0
(3) Ethanol 10.0
(4) Glycerin 5.0
(5) Polyethylene glycol (average molecular weight 1000) 2.0
(6) Willow milkweed extract [Preparation method 1] 1.0
(7) Milkweed extract [Preparation method 2] 1.0
(8) Fragrance 0.1
Production method: (2) and (3) are mixed, heated to 80 ° C, and then dissolved in (1) heated to 80 ° C. After uniformly dissolving, (4) to (5) are added, and cooling is started while stirring. Cool to 40 ° C., add (6) to (8) and mix uniformly.
[0043]
Production method: The oil phase components (1) to (6) are mixed and heated and dissolved at 75 ° C. On the other hand, the aqueous phase components (7) to (10) are dissolved by heating at 75 ° C., the oil phase component is added, and the mixture is emulsified with a homomixer. Cooling is started after the emulsification is completed, and the components (11) to (13) are added at 40 ° C. and mixed uniformly.
[0044]
[Example 18] Bath agent (1) Fragrance 0.3 (% by weight)
(2) Milkweed extract [Preparation method 1] 0.5
(3) Milkweed extract [Preparation method 3] 0.5
(4) Sodium bicarbonate 50.0
(5) Sodium sulfate 48.7
Production method: (1) to (5) are mixed uniformly.
[0045]
Examples 1 to 5 of the present invention were subjected to use tests to evaluate the effect of improving wrinkles, tarmi and skin firmness. At that time, in Example 1, the blended milkweed extract was replaced with purified water, and a use test was simultaneously conducted as Comparative Example 1.
[0046]
For each sample, a group of 20 male and female panelists in their 50s and 60s who have noticeable symptoms such as wrinkles, tarmi, and reduced skin firmness, were used blindly for 1 month, and the skin condition before and after use Changes were observed and evaluated. As indices of skin symptoms, wrinkles, tarmi, and skin firmness were evaluated in three stages, “improved”, “slightly improved”, and “no change”, and Table 4 shows the number of panelists that obtained each evaluation.
[0047]
[Table 4]
[0048]
As shown in Table 4, the wrinkles, tarmi, and skin firmness were not improved in more than half of the panelers in the comparative example use group not containing the extract of the plant of the genus Asclepias , but the plant of the genus Asclepias In the example use group containing the extract, a clear improvement was observed in 60% or more of the panelists.
[0049]
As mentioned above, in the Example of this invention, it had the effect excellent in improvement of the aging symptom, such as a wrinkle, a talmi, and skin elasticity, compared with the conventional comparative example.
[0050]
【The invention's effect】
As described above in detail, according to the present invention, a cell activator and an antioxidant having excellent effects could be obtained. Moreover, the skin external preparation which has the effect excellent in prevention and improvement of the skin aging symptom was able to be obtained by mix | blending the cell activator and antioxidant which were such as an active ingredient.
Claims (4)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2002301489A JP3665049B2 (en) | 2002-10-16 | 2002-10-16 | Cell activator, antioxidant, and skin external preparation |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2002301489A JP3665049B2 (en) | 2002-10-16 | 2002-10-16 | Cell activator, antioxidant, and skin external preparation |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2004137161A JP2004137161A (en) | 2004-05-13 |
JP3665049B2 true JP3665049B2 (en) | 2005-06-29 |
Family
ID=32449814
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2002301489A Expired - Fee Related JP3665049B2 (en) | 2002-10-16 | 2002-10-16 | Cell activator, antioxidant, and skin external preparation |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP3665049B2 (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1763358A4 (en) * | 2004-04-07 | 2010-03-24 | Univ Rutgers | Appetite-suppressing compositions and methods |
CN104523792B (en) * | 2014-12-24 | 2017-11-07 | 暨南大学 | A kind of milkweed latex extract rich in cardiac glycoside and preparation method and application |
-
2002
- 2002-10-16 JP JP2002301489A patent/JP3665049B2/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
JP2004137161A (en) | 2004-05-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5557414B2 (en) | Moisturizer, cell activator, and whitening agent | |
JP3754025B2 (en) | Cell activator, antioxidant, and skin external preparation | |
JP3665049B2 (en) | Cell activator, antioxidant, and skin external preparation | |
JP4748962B2 (en) | Moisturizer, cell activator, whitening agent, and antioxidant | |
JP3734222B2 (en) | External preparation for skin, cell activator, and antioxidant | |
JP2004137166A (en) | Skin care preparation for external use, cell activator and antioxidant | |
JP2003267851A (en) | Skin care preparation | |
JP3627862B2 (en) | Topical skin preparation | |
JP4245862B2 (en) | Topical skin preparation | |
JP4515423B2 (en) | Moisturizer, cell activator, and antioxidant | |
JP4295473B2 (en) | Topical skin preparation | |
JP4050727B2 (en) | Moisturizer, cell activator, whitening agent, antioxidant, and skin external preparation | |
JP4994684B2 (en) | Moisturizer, cell activator, and antioxidant | |
JP2003137766A (en) | Collagen production promoter for external use for skin, and hyaluronic acid production promoter for external use for skin | |
JP4480366B2 (en) | Cell activator, whitening agent, hyaluronic acid production promoter, collagen production promoter, antioxidant, and skin external preparation | |
JP4515425B2 (en) | Moisturizer, cell activator, whitening agent, and antioxidant | |
JP4291647B2 (en) | Cell activator, whitening agent, collagen production promoter, antioxidant, and skin external preparation | |
JP4931452B2 (en) | Moisturizer, cell activator, whitening agent, and antioxidant | |
JP3543306B2 (en) | Cell activator and antioxidant | |
JP4018605B2 (en) | Cell activating agent, whitening agent, antioxidant, and skin external preparation | |
JP4916150B2 (en) | Moisturizer, cell activator, antioxidant, and skin external preparation, | |
JP4050674B2 (en) | Cell activator, whitening agent, hyaluronic acid production promoter, elastin production promoter, antioxidant, and external preparation for skin | |
JP4033349B2 (en) | Cell activator, whitening agent, hyaluronic acid production promoter, decorin production promoter, antioxidant, and external preparation for skin | |
JP5009544B2 (en) | Moisturizer, cell activator, antioxidant, and collagenase inhibitor | |
JP4936754B2 (en) | Moisturizer, cell activator, and collagenase inhibitor |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20041125 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20041201 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20050131 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20050308 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20050330 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 3665049 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20090408 Year of fee payment: 4 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20100408 Year of fee payment: 5 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20100408 Year of fee payment: 5 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110408 Year of fee payment: 6 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130408 Year of fee payment: 8 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130408 Year of fee payment: 8 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20140408 Year of fee payment: 9 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
LAPS | Cancellation because of no payment of annual fees |