JP6040233B2 - 孵化液から美容組成物を製造する方法および皮膚の美容上の外観を改善するためのその使用 - Google Patents
孵化液から美容組成物を製造する方法および皮膚の美容上の外観を改善するためのその使用 Download PDFInfo
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- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
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Description
a)サケ科の卵を、最小量の水(例えば、前記卵の量と同量以下)に懸濁させる工程;
b)当該卵の同期化させた急速孵化を、(好ましくは、95%を超える胚について、2時間未満に孵化が完了するように)誘発する工程;
c)任意に、孵化した卵を濾過し、孵化液を得る工程;および
d)前記孵化液を濾過し、ポリペプチドおよび/または組成物を得る工程
を含むサケ科孵化液(例えば、サケ孵化液)から本明細書に記載の美容組成物を調製する方法であって、
前記孵化液を濾過する工程が、少なくとも下記工程:
(i)孔径が5μm以上、好ましくは5〜15μm、特に好ましくは孔径7μmのフィルターを用いて前記孵化液を濾過し、その濾液を回収する工程;
(ii)孔径が0.30〜0.60μm、好ましくは孔径0.35〜0.55μm、特に好ましくは0.40〜0.50μm、最も好ましくは0.45μmのフィルターを用いて、工程(i)の前記濾液を濾過し、その濾液を回収する工程;
(iii)工程(ii)の前記濾液中の水を、医薬上許容される緩衝液と交換する工程;
(iv)孔径が0.15〜0.30μm、好ましくは孔径0.22μmのフィルターを用いて、工程(iii)において得られた溶液を濾過し、その濾液を回収する工程;および
(v)工程(iv)の前記濾液から、前記美容組成物を調製する工程
を含む方法、を提供する。
前記美容組成物の0.0001〜10%(重量比)(または10〜40%以下)、例えば、前記方法に従って調製された美容組成物の0.0001%〜5%、0.0001%〜3%、0.0001%〜2%、0.0001%〜1%、0.0001%〜0.5%、0.0001%〜0.1%(重量比)の範囲内で(組合せで)存在していてもよく、或いは、以下記載するように、例えば、希釈後の状態であってもよい。従って、サケ科孵化液から抽出可能な個々のポリペプチドまたはポリペプチドの一部は、美容組成物の1x10-6〜10%(重量比)の範囲内で存在していてもよい。実施の形態によっては、サケ科孵化液から抽出可能な当該個々のポリペプチドまたはポリペプチドの一部は、美容組成物の1x10-6〜5%(重量比)、例えば、美容組成物の1x10-6〜4%、1x10-6〜3%、1x10-6〜2%、1x10-6〜1%、1x10-6〜0.5%、1x10-6〜0.1%または1x10-6〜0.01%(重量比)の範囲内で存在していてもよい。
図1は、本発明の孵化液組成物を用いて治療した被験者の、治療前(ベースライン)、治療の2週間後および12週間後の写真を示す。老化皮膚の様々な徴候の低減が、2週間後および12週間後のいずれにおいても明らかである。記載されている値は、35名の参加者に関する平均変化を示す。
図2は、本発明の孵化液組成物を用いた治療の2週間後および12週間後に見られた小皺および皺の低減を強調する、図1の被験者のクローズアップ写真を示す。
図3は、本発明の孵化液組成物を用いて治療した被験者の、治療前(ベースライン)および治療の12週間後のクローズアップ写真を示す。円で囲んだ部位は、治療の12週間後の明瞭な皺の低減を示す。
図4は、顔面の両側に関する触覚的/視覚的臨床評価に基づき、老化の様々な徴候が改善されたとみなされる被験者の割合を示す棒グラフである。
図5は、水、5%グリコール酸、1mU/mlのブロメラインまたは1%孵化液組成物で12時間治療したヒトの再構成皮膚の走査電子顕微鏡写真(倍率400倍、5000倍および15000倍)を示す。
図6は、水、5%グリコール酸、1mU/mlのブロメラインまたは1%孵化液組成物で48時間治療したヒトの再構成皮膚の走査電子顕微鏡写真(倍率400倍、5000倍および15000倍)を示す。
図7は、5%グリコール酸、1mU/mlのブロメラインまたは1%孵化液組成物で12時間または24時間治療したまたは未治療のヒトの再構成皮膚の断面の光学マイクロ写真を示す。矢印(A)は、細胞増殖および分化を示し、矢印(B)は、より高い密度の顆粒層およびより高い濃度の層状顆粒を示す。
前記組成物を、サケ孵化液から調製した。孵化液のタンパク質濃度を高めるため、サケ卵は、孵化に先立って最小量の水に移した。高度に同期した孵化は、高(室)温により、または脱酸素化(オッペン−バルンツンら、1990年、アクアカルチャー、86巻、417〜430頁)により誘発することができ、未精製のポリペプチドおよびポリペプチドの一部の高度に濃縮された調製物が少量得られる。孵化は、95%を超える胚について、2時間未満に完了するものとする。
前記孵化液組成物を、実施例1に記載の如く調製した。前記組成物を、試験に際し2つの活性スキンローションである、1%および3%スキンローション[容量比](ローション単位容量当りの組成物の総容量)として調製し、有効成分、つまり、孵化液組成物を含んでいない対照スキンローションと比較した。前記スキンローションは、水中油型(O/W)乳剤であった。この油相は、組成物全体の9%に相当し、水素添加レシチンで乳化した。
101名の女性被験者がこの試験への参加を完了した(3つの治療、すなわち、1つのプラセボと、前記孵化液組成物を異なる濃度で含む2つの組成物に関して;N>30)。
被験者は、40〜65歳の健康な女性であり、臨床試験番号と共に使用した場合、その試験における各被験者を一意的に特定する3桁の番号を割り当てた。この番号は、この試験の全期間を通して被験者に使用され、実験の匿名性を維持した。
<試験対象被験者基準>
1.健康および適格性アンケートにより判断した、全般的に健康である、40〜65歳までの女性。
2.試験の期間中、試験の必要条件に従い、協力および参加しようとする意志並びにいかなる有害な症状も直ちに報告しようとする意志。
3.スコア3〜7の修正グリフィス評価尺度(modified Griffith’s grading scale)に相応する、臨床的に判断した軽度から中等度の顔面における光損傷(小皺、皺、色素沈着過剰、緩みおよび荒れ)。
4.テスト部位の評価を損なう、または試験への参加により被験者の健康リスクが増える可能性のあるいかなる疾病状態または物理的な顔面皮膚疾患(例えば、アトピー性皮膚炎、湿疹、乾癬、脂漏性皮膚炎)もないこと。
5.試験の期間中、長時間に渡る太陽への露出および日焼けマシーンの全使用を避けようとする意志。サングラスを含め保護用の衣服を着用し、午前10時〜午後4時まで太陽への露出を避けるよう特別な注意を払うものとする。
6.試験の期間中、全ての通常のブランドのカラー化粧品、洗顔料、化粧水(該当する場合)およびメイク落としの使用を継続しようとする意志。各個人は、割り当てられた試験物質以外のいかなる抗老化製品または皮膚美白製品も使用を控えなければならなかった。
7.指定された各クリニック訪問に先立ち、少なくとも20分前に、全ての化粧を落とそうとする意志。試験訪問が終わるまで、他の局所製品を、顔面または目の周りに適用しないものとした。被験者が化粧を全て落とすことなく到着した場合は、クリニックで残りの化粧を落とし、処置に先立って少なくとも20分待たなければならなかった。
8.ホルモン補充療法を受けているか、または受胎調節用ホルモンを服用している個人は、試験開始に先立って少なくとも1カ月間は、安定したレジメンに従っていなければならず、また、試験の期間中、この薬剤を変えることなく続ける意志があることが必要であった。試験開始の時点でホルモン補充療法(HRT)を受けていないまたはホルモンを服用していない個人は、試験過程の間、使用を開始しない意志があることが必要であった。
9.試験の必要条件に従い、協力および参加しようとする意志並びにいかなる有害な症状も直ちに報告しようとする意志。
1.任意のパーソナルケア製品に対する不耐性またはアレルギーの既往歴のある個人。
2.試験エントリーの前30日未満に、任意の処方薬または市販の皮膚美白製品を使用した個人。
3.検査する研究者が、参加に不適当とみなした皮膚の疾患および/または疾病を有していた個人。
4.授乳していた、妊娠していた、またはこの試験期間中に妊娠を予定していた個人。
5.試験エントリーの前30日未満に、何らかの抗老化、抗皺、局所的抗酸化剤を日常的に使用していた個人。
6.試験開始前6カ月以内に酵素によるフェイシャルスキントリートメントをした個人。
7.レチン−A(Retin−A;登録商標)、レチン−Aマイクロ(Retin−A Micro;登録商標)、レノバ(Renova;登録商標)、アビタ(Avita;登録商標)、タゾラック(Tazorac;登録商標)、Avage(登録商標)若しくはディフェリン(Differin;登録商標)または他の局所レチノイドの試験開始前3カ月以内における使用、あるいは、アキュテイン(Accutane)または経口レチノイドの過去6カ月以内における服用。
8.α−、β−またはポリ−ヒドロキシ酸(サリチル酸およびラクハイドリン(Lachydrin)を含む)、レチノール若しくはレチノールの誘導体を含有する製品、または他の「抗老化」製品の試験開始前30日以内における顔面への日常的使用。
9.治療前3カ月以内または試験期間中に顔の皮膚擦傷術またはケミカルピーリングトリートメントを受けた個人。
10.治療前6カ月以内または試験期間中に、治療エリア内の治療部位に、ライトRF(light RF)または他の装置を使用した治療を受けた個人。
11.治療前9カ月以内または試験期間中に、治療部位にボトックス、コラーゲン、脂肪注入またはその他の注射物質または注入物質を用いたオーグメンテーション法を受けた個人。
12.この試験の開始前12カ月以内にリサーフェシング治療、フェースリフトまたは目若しくはまぶたの手術を受けた個人。
13.顔面に既存のおよび/若しくは潜伏皮膚疾患(例えば、白斑、アトピー性皮膚炎、乾癬、酒さ、湿疹、脂漏性皮膚炎、重度の表皮剥離など)または研究者の見解において、試験の結果に干渉し得る医学的疾患/疾病を有していた個人。
14.免疫抑制剤/免疫不全障害(ヒト免疫不全ウイルス(HIV)または後天性免疫不全症候群(AIDS)を含む)の既往歴を有していたか、またはその時点で免疫抑制薬を使用していた個人。
15.任意の他の臨床使用試験(パッチ試験は容認)に参加していた個人。
16.糖尿病、高血圧症、甲状腺機能亢進症または甲状腺機能低下症といった抑制不可能な疾病を有していた個人。複数の健康上の疾患を有していた数人に関しては、たとえ、それらの疾患が、食事制限、薬剤などにより抑制されるとしても、参加を拒否した。
17.この試験への参加前28日以内に何らかの臨床試験に参加した個人。
二重盲検プラセボ対照臨床試験を行い、軽度から中等度の光損傷を受けた、すなわち、老化した顔面皮膚を有する女性における局所皮膚治療の有効性を評価した。この試験の期間は、12週間であり、ベースライン、第2週目、第6週目および第12週目に来院日を設けた。視覚的評価、計測手段、VISIA−CRデジタル写真および被験者による自己評価アンケートを用いて有効性を評価した。
熟練した臨床評価者が、下記に示すパラメーターに関して、顔面の右側および左側を評価した。修正グリフィス尺度を使用し、0=なし、1〜3=軽度、4〜6=中等度および7〜9=重度とした。皮膚の状態をよりよく描写する上で必要な場合は、半ポイントを使用した。
・小皺
・皺
・色素沈着過剰
・緩み
・くすみ/艶が無い(透明度)
・触覚的粗さ
・紅斑
・乾燥/鱗屑
・ヒリヒリ感/チクチク感
・かゆみ
・突っ張り感/乾燥感
顔面の右側および左側のVISIA−CR画像を撮影した。被験者は、髪は後方に流し、宝石類は外し、目は閉じ、フレーム内の中央に位置させ、またニュートラルな表情をした状態で撮影した。
計器測定に先立ち、被験者は、少なくとも20分間、クリニックの環境条件に平衡させた。環境条件は、試験訪問中、1時間ごとに記録した。この時間の間に、被験者は評価され、アンケートに記入し、なお且つ/またはVISIA−CR画像撮影が行われた。
全被験者は、全訪問時において、キュートメーター測定を行った。キュートメーターを使用し、皮膚の粘弾性特性(すなわち、伸張性および弾力性)を評価した。当該装置は、皮膚の小さな部位に真空を適用し、皮膚の弾性応答(開口部内および開口部外への皮膚の動き)を光学的手法により測定する。
被験者は、顔の右側および左側における、自己の顔の皮膚外観および状態を被験者がどのように知覚しているかを記述する質問を含む、皮膚自己評価アンケートに記入した。
<小皺>
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において、プラセボ(変化率4.58%)に比べて、小皺の低減(例えば、変化率5.59%(1%溶液)および5.65%(3%溶液))を示した。この小皺の低減は、第6週目(例えば、14.34%(1%溶液)、14.86%(3%溶液)および8.98%(プラセボ))および第12週目(例えば、23.43%(1%溶液)、25.99%(3%溶液)および14.68%(プラセボ))でも続いた。図1および図2は、小皺が28%低減した被験者を示す。
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において、プラセボ(変化率0.70%)に比べて、皺の低減(例えば、変化率2.15%(1%溶液)および1.75%(3%溶液))を示した。この皺の低減は、第6週目(例えば、6.13%(1%溶液)、7.32%(3%溶液)および3.70%(プラセボ))および第12週目(例えば、14.72%(1%溶液)、15.15%(3%溶液)および9.57%(プラセボ))でも続いた。図1は、皺が12.5%低減した被験者を示す。図3は、皺が26.32%低減した被験者を示す。
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において、プラセボ(変化率0.40%)に比べて、色素沈着過剰の低減(例えば、変化率2.11%(1%溶液)および2.68%(3%溶液))を示した。この色素沈着過剰の低減は、第6週目(例えば、5.61%(1%溶液)、7.91%(3%溶液)および3.16%(プラセボ))および第12週目(例えば、10.53%(1%溶液)、15.35%(3%溶液)および5.73%(プラセボ))でも続いた。図1は、12週間後、年齢による染みの色素沈着が15%低減した被験者を示す。
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において、プラセボ(変化率0.87%)に比べて、緩みの低減(例えば、変化率2.64%(1%溶液)および1.62%(3%溶液))を示した。この緩みの低減は、第6週目(例えば、6.33%(1%溶液)および6.61%(3%溶液)、2.51%(プラセボ))および第12週目(例えば、10.55%(1%溶液)および11.33%(3%溶液)、5.18%(プラセボ))でも続いた。図1は、12週間後、緩み(たるみ)が7.69%低減した被験者を示す。
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において、プラセボ(変化率10.67%)に比べて、皮膚透明度の向上(例えば、変化率12.95%(1%溶液)および16.00%(3%溶液))を示した。この向上は、第6週目(例えば、29.26%(1%溶液)、28.50%(3%溶液)および19.07%(プラセボ))および第12週目(例えば、37.17%(1%溶液)、39.18%(3%溶液)および26.72%(プラセボ))でも続いた。図1は、くすみが33.33%低減した被験者を示す。
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において、プラセボ(変化率13.38%)に比べて、皮膚の触覚的粗さの低減(例えば、変化率16.51%(1%溶液)および20.24%(3%溶液))を示した。この改善は、第6週目(例えば、24.77%(1%溶液)、26.65%(3%溶液)および16.79%(プラセボ))でも続いたが、第12週目(例えば、26.61%(1%溶液)、29.19%(3%溶液)、および15.79%(プラセボ))での更なる低減はなかった。図1は、触覚的粗さが12.5%低減した被験者を示す。
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において、プラセボ(変化率64.71%)に比べて、皮膚の乾燥/鱗屑の低減(例えば、変化率72.09%(1%溶液)および100.00%(3%溶液))を示した。しかしながら、第6週目(例えば、86.05%(1%溶液)、84.62%(3%溶液)および100.00%(プラセボ))および第12週目(例えば、90.70%(1%溶液)、100.00%(3%溶液)および89.47%(プラセボ))では、プラセボに比べて更なる低減はなかった。
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において、TEWLの低減を示したが、これは、プラセボ(変化率17.26%)に比べて、プラセボとの明らかな違いはなかった(例えば、変化率15.35%(1%溶液)および14.53%(3%溶液))。しかしながら、第6週目(例えば、29.46%(1%溶液)、26.66%(3%溶液)および22.96%(プラセボ))および第12週目(例えば、37.46%(1%溶液)、40.04%(3%溶液)、および34.21%(プラセボ))においては、プラセボよりも大きく更なる低減が見られた。
有効組成物の1つを含むスキンローションで治療した被験者は、第2週目において皮膚の伸張性の改善を示したが、これは、プラセボ(変化率10.82%)と比べて、プラセボとわずかに異なるのみであった(例えば、変化率16.18%(1%溶液)および17.21%(3%溶液))。第6週目において、3つの治療間における明らかな違いはなかった(例えば、18.04%(1%溶液)、17.18%(3%溶液)および19.90%(プラセボ))が、第12週目において、有効成分を含む組成物を用いた皮膚治療に関して、更なる改善が見られ、これは、プラセボよりも大きく改善した(例えば、31.84%(1%溶液)、33.57%(3%溶液)、および16.48%(プラセボ))。
ヒトの生体外再構成表皮は、合成培地において、気液界面での不活性なポリカーボネートフィルター上で培養された正常ヒトケラチノサイトから成る。この生体外モデルは、ヒトの生体内表皮のそれと組織学的に類似している。
1%孵化液組成物;
5%グリコール酸(AHA);
1mU/mlブロメライン(フルーツ酵素);または
dH2O(対照)。
Claims (20)
- 哺乳類動物に美容組成物を投与することを含む、当該動物の皮膚における皺、小皺、色素沈着過剰、緩み、乾燥皮膚、鱗屑および/もしくは経表皮水分喪失の広がりまたは美容上の外観を低減または予防する方法であって、前記組成物が、少なくとも下記工程:
a)最小量の水にサケ科の卵を懸濁させる工程;
b)前記卵の同期化させた急速孵化を、誘発する工程;
c)孵化した卵を濾過し、孵化液を得る工程;および
d)前記孵化液を濾過し、前記組成物を得る工程
を含む方法により得られたか、または得ることが可能であり、
前記孵化液を濾過する工程が、少なくとも下記工程:
(i)孔径が5μm以上のフィルターを用いて、前記孵化液を濾過し、その濾液を回収する工程;
(ii)孔径が0.30〜0.60μmのフィルターを用いて、工程(i)の前記濾液を濾過し、その濾液を回収する工程;
(iii)排除サイズが15kDa未満のフィルターを使用するダイアフィルトレーションを用いて行うことにより、工程(ii)の前記濾液中の水を、医薬上許容される緩衝液と交換する工程;
(iv)孔径が0.15〜0.30μmのフィルターを用いて、工程(iii)の前記溶液を濾過し、その濾液を回収する工程;および
(v)工程(iv)の前記濾液から前記美容組成物を調製する工程
を含む、方法。 - 前記サケ科の卵が、サケ卵である請求項1に記載の方法。
- 95%を超える胚について、2時間未満に孵化が完了する請求項1または2に記載の方法。
- a)工程(i)における前記フィルターの孔径が5〜15μmであり;
b)工程(ii)における前記フィルターの孔径が0.35〜0.55μmであり;および/または
c)工程(v)における前記フィルターの孔径が0.22μmである、請求項1〜3のいずれか一項に記載の方法。 - a)工程(i)における前記フィルターの孔径が7μmであり;および/または
b)工程(ii)における前記フィルターの孔径が0.45μmである請求項4に記載の方法。 - 前記サケ科の卵が、タイセイヨウサケまたはタイヘイヨウサケのものである、請求項1〜5のいずれか1つに記載の方法。
- 前記皮膚が老化皮膚である、請求項1〜6のいずれか1つに記載の方法。
- 前記動物の皮膚に潤いを与える、請求項1〜7のいずれか1つに記載の方法。
- 前記美容組成物が、前記皮膚への局所投与用である、請求項1〜7のいずれか1つに記載の方法。
- 請求項1〜9のいずれか1つに記載の方法であって、前記美容組成物は、製品、素材または道具に、被覆、含浸または化学結合される方法。
- 請求項1〜10のいずれか1つに記載の方法であって、皺および/もしくは緩みの広がりまたは美容上の外観を低減または予防する方法。
- 哺乳類動物の皮膚の皺、小皺、色素沈着過剰、緩み、乾燥皮膚、鱗屑および/若しくは経表皮水分喪失の広がりまたは美容上の外観の低減あるいは予防に使用するための請求項1〜6のいずれか1つに記載の美容組成物。
- 前記皮膚が、請求項7に記載の通りであり、前記皮膚の美容上の外観が、請求項8に記載の通りであり、および/または前記投与が、請求項9に記載の通りである、請求項12に記載の美容組成物。
- 皺および/または緩みの広がりまたは美容上の外観を低減または予防するための請求項12または13に記載の美容組成物。
- 哺乳類動物の皮膚の皺、小皺、色素沈着過剰、緩み、乾燥皮膚、鱗屑および/若しくは経表皮水分喪失の広がりまたは美容上の外観を低減あるいは予防するための薬剤製造における、請求項1〜6のいずれか1つに記載の美容組成物の使用。
- 前記皮膚が、請求項7に記載の通りであり、前記皮膚の美容上の外観が、請求項8に記載の通りであり、および/または前記投与が、請求項9に記載の通りである、請求項15に記載の美容組成物の使用。
- 前記薬剤が、皺および/もしくは緩みの広がりまたは美容上の外観を低減または予防するための薬剤である請求項15または16に記載の使用。
- 少なくとも下記工程:
a)最小量の水にサケ科の卵を懸濁させる工程;
b)前記卵の同期化させた急速孵化を、誘発する工程;
c)孵化した卵を濾過し、孵化液を得る工程;および
d)前記孵化液を濾過し、美容組成物を得る工程
とを含む、前記美容組成物を調製する方法であって、
前記孵化液を濾過する工程が、少なくとも下記工程:
(i)孔径が5μm以上のフィルターを用いて、前記孵化液を濾過し、その濾液を回収する工程;
(ii)孔径が0.30〜0.60μmのフィルターを用いて、工程(i)の前記濾液を濾過し、その濾液を回収する工程;
(iii)排除サイズが15kDa未満のフィルターを使用するダイアフィルトレーションを用いて行うことにより、工程(ii)の前記濾液中の水を、医薬上許容される緩衝液と交換する工程;
(iv)孔径が0.15〜0.30μmのフィルターを用いて、工程(iii)の前記溶液を濾過し、その濾液を回収する工程;および
(v)工程(iv)の前記濾液から前記美容組成物を調製する工程
を含む、方法。 - 前記サケ科の卵が、サケ卵である請求項18に記載の方法。
- 95%を超える胚について、2時間未満に孵化が完了する請求項18または19に記載の方法。
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GB201110783D0 (en) | 2011-06-24 | 2011-08-10 | Aqua Bio Technology Asa | Methods and uses |
BR112014017913A8 (pt) * | 2012-01-23 | 2017-07-11 | Restorsea Llc | Cosmético na forma de uma emulsão, kit e processo para melhora da aparência da pele |
GB201223330D0 (en) | 2012-12-21 | 2013-02-06 | Aqua Bio Technology Asa | Products, methods and uses |
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- 2012-06-25 CN CN201280031250.0A patent/CN103732292B/zh active Active
- 2012-06-25 RU RU2014102049/15A patent/RU2558848C1/ru active
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CA2839641A1 (en) | 2012-12-27 |
US10987302B2 (en) | 2021-04-27 |
NZ620189A (en) | 2015-12-24 |
CN103732292A (zh) | 2014-04-16 |
RU2558848C1 (ru) | 2015-08-10 |
CA2839641C (en) | 2019-04-23 |
KR101932307B1 (ko) | 2019-03-20 |
JP2014517059A (ja) | 2014-07-17 |
RU2014102049A (ru) | 2015-07-27 |
PT2723453T (pt) | 2017-04-03 |
DK2723453T3 (en) | 2017-04-10 |
ZA201400559B (en) | 2015-07-29 |
CN103732292B (zh) | 2017-03-15 |
ES2621833T3 (es) | 2017-07-05 |
US20190350837A1 (en) | 2019-11-21 |
WO2012175743A2 (en) | 2012-12-27 |
AU2012273931B2 (en) | 2016-12-15 |
EP2723453B1 (en) | 2017-01-18 |
WO2012175743A3 (en) | 2013-08-08 |
US10328021B2 (en) | 2019-06-25 |
EP2723453A2 (en) | 2014-04-30 |
HUE031610T2 (en) | 2017-07-28 |
KR20140072016A (ko) | 2014-06-12 |
US20140220088A1 (en) | 2014-08-07 |
GB201110783D0 (en) | 2011-08-10 |
BR112013033212A2 (pt) | 2016-09-06 |
AU2012273931A1 (en) | 2014-02-06 |
PL2723453T3 (pl) | 2017-08-31 |
HK1197205A1 (en) | 2015-01-09 |
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