JP2014501777A - アロエ多糖体の組成物及び方法 - Google Patents
アロエ多糖体の組成物及び方法 Download PDFInfo
- Publication number
- JP2014501777A JP2014501777A JP2013548389A JP2013548389A JP2014501777A JP 2014501777 A JP2014501777 A JP 2014501777A JP 2013548389 A JP2013548389 A JP 2013548389A JP 2013548389 A JP2013548389 A JP 2013548389A JP 2014501777 A JP2014501777 A JP 2014501777A
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- JP
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- Prior art keywords
- aloe
- polymannan extract
- polysaccharide
- daltons
- powder
- 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.)
- Granted
Links
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Classifications
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0084—Guluromannuronans, e.g. alginic acid, i.e. D-mannuronic acid and D-guluronic acid units linked with alternating alpha- and beta-1,4-glycosidic bonds; Derivatives thereof, e.g. alginates
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Abstract
Description
機器:HPLCシステムは、Hitachi社製L-7100ポンプ、及びWaters社製410示差屈折計と対になった7250オートサンプラーである。SECは、カラムヒーター内、70℃で動作する、Tosoh Biosep社製G6000 PWXL TSK Gel 30cm×7.8mmである。分子量標準物質は、Sigma社製−2,000,000ダルトン、1,000,000ダルトン、480,000ダルトン、66,000ダルトン及び180ダルトン(グルコース)である。移動相は、流速0.70mL/分の脱イオン水である。注射体積は10uLである。SEC法がPughらにより記述されている(Pugh N., Ross S.A., ElSohly M.A., and Pasco, D.S. (2001). Characterization of Aloeride, a new high-molecular weight polysaccharide from Aloe vera with potent immunomodulatory activity. J Agr. Food Chem., 49, 1030-1034)。
United States Patent No. 7,196,072: High Molecular Weight Polysaccharide Fraction From Aloe Vera with Immunostimulatory Activity
United States Patent No. 6,083,508: Method of Processing Aloe Leaves
United States Patent Application No. 2006/0084629: Immune System Activating Formula Composed of Selected Long Chain Polysaccharides From Natural Sources
1 Pugh N., Ross S.A., ElSohly M.A., and Pasco, D.S. (2001). Characterization of Aloeride, a new high-molecular weight polysaccharide from Aloe vera with potent immunomodulatory activity. J Agr. Food Chem., 49, 1030-1034
Claims (48)
- ポリマンナン抽出物の微粉末の調製方法であって、
所定量のフリーズドライされたアロエ粉末を秤量するステップであって、前記量が水分含有量に対して補正されるステップと、
前記フリーズドライされたアロエ粉末を脱イオン水に溶解して、溶液を形成するステップと、
有機溶媒を前記溶液に添加して第1の混合物を形成するステップであって、前記有機溶媒と前記脱イオン水との比が少なくとも2.5:1であるステップと、
前記第1の混合物を少なくとも8時間静置させるステップと、
所定体積の上清を前記第1の混合物から抜き取り、過剰体積の前記上清溶液を添加して、第2の混合物を形成するステップと、
前記第2の混合物を遠心分離するステップと、
第2の混合物中における沈殿物の存在を観察するステップと、
第2の混合物中において任意の沈殿物が観察された場合に、追加量の前記有機溶媒を第1の混合物に添加するステップと、
吸い上げることによって第1の混合物の上清をデカンテーションするステップであって、前記デカンテーションが、第2の混合物中において沈殿物が観察されない場合にのみ実施されるステップと、
減圧下、濾紙及び吸引漏斗を使用して第1の混合物から前記沈殿物を濾過するステップと、
削り取ることによって前記吸引漏斗から前記ポリマンナン抽出物の粉末を回収するステップと、
キャップをした凍結乾燥フラスコ中の前記ポリマンナン抽出物の粉末を、冷凍庫内に少なくとも8時間置くステップと、
前記ポリマンナン抽出物の冷凍粉末を、凍結乾燥機内で凍結乾燥させるステップと、
前記ポリマンナン抽出物の凍結乾燥粉末を、粉砕機内で細粒質に粉砕するステップと
を含む、前記方法。 - ポリマンナン抽出物の微粉末を、秤量し、ラベル付けし、容器内に保存するステップをさらに含む、請求項1に記載の方法。
- フリーズドライされたアロエ粉末が、アロエ・ベラ、キダチアロエ、アヤニシキ、タカロカイ、アロエ・ニエリエンシス、チヨダニシキ(Aloe variegate)、ホンロカイ及びアロエ・ウィルディからなる群から選択されるアロエ種に由来する、請求項1に記載の方法。
- 有機溶媒が、メタノール、エタノール、イソプロピルアルコール及びプロパノールからなる群から選択される、請求項1に記載の方法。
- フリーズドライされたアロエ粉末がアロエ多糖体を含み、前記アロエ多糖体が、1又は2以上の短鎖、中鎖、長鎖、極長鎖多糖体、又はそれらの任意の組合せを含む、請求項1に記載の方法。
- アロエ多糖体が、グルコース、マンノース、アラビノース及びガラクトースからなる群から選択される単糖をさらに含む、請求項5に記載の方法。
- アロエ多糖体が、11,500ダルトン〜10,000,000ダルトン超の範囲の分子量を有する、請求項5に記載の方法。
- フリーズドライされたアロエ粉末が、少なくとも25%のアロエ多糖体を有する、請求項1に記載の方法。
- フリーズドライされたアロエ粉末が、25%、30%、40%、50%、60%、70%、80%、90%又は95%のアロエ多糖体を有する、請求項1に記載の方法。
- フリーズドライされたアロエ粉末が、66,000ダルトンの分子量を有するアロエ多糖体を少なくとも14%含む、請求項1に記載の方法。
- フリーズドライされたアロエ粉末が、480,000ダルトンの分子量を有するアロエ多糖体を少なくとも9%含む、請求項1に記載の方法。
- フリーズドライされたアロエ粉末が、1,000,000ダルトンの分子量を有するアロエ多糖体を少なくとも3.5%含む、請求項1に記載の方法。
- フリーズドライされたアロエ粉末が、2,000,000ダルトンの分子量を有するアロエ多糖体を少なくとも2.4%含む、請求項1に記載の方法。
- フリーズドライされたアロエ粉末中における2,000,000ダルトンの分子量を有するアロエ多糖体の量が、1.32%〜6.36%の範囲である、請求項1に記載の方法。
- フリーズドライされたアロエ粉末中における1,000,000ダルトンの分子量を有するアロエ多糖体の量が、2.55%〜3.89%の範囲である、請求項1に記載の方法。
- フリーズドライされたアロエ粉末中における480,000ダルトンの分子量を有するアロエ多糖体の量が、63.85%〜73.36%の範囲である、請求項1に記載の方法。
- フリーズドライされたアロエ粉末が、グルコース、有機酸、乳酸、リンゴ酸、クエン酸及びアスパラギン酸からなる群から選択される1又は2以上の残留低分子量種を含有し得る、請求項1に記載の組成物。
- 1又は2以上の残留低分子量種が、14%〜24%の範囲の量で存在する、請求項17に記載の方法。
- ポリマンナン抽出物の微粉末が、医薬組成物の調製において使用される、請求項1に記載の方法。
- 医薬組成物が、白血病及びリンパ腫、前立腺がん、乳がん並びに結腸がんからなる群から選択される1又は2以上の悪性腫瘍の治療において、並びに1又は2以上の免疫障害の治療のために使用される、請求項19に記載の方法。
- 脱イオン水に溶解された所定量の微細なポリマンナン抽出物と、
1又は2以上の医薬品保存剤と
を含む、ポリマンナン抽出物の注射用製剤。 - 注射用製剤が無菌である、請求項21に記載の組成物。
- 1又は2以上の医薬品保存剤が、パラベン、安息香酸及びそれらの塩、水銀剤、第4級アンモニウム塩、ベンジルアルコール及び他の関連アルコール、並びにフェノールからなる群から選択される、請求項21に記載の組成物。
- 保存剤がベンジルアルコールである、請求項21に記載の組成物。
- ポリマンナン抽出物がアロエ多糖体を含み、前記アロエ多糖体が、1又は2以上の短鎖、中鎖、長鎖、極長鎖多糖体、又はそれらの任意の組合せを含む、請求項21に記載の組成物。
- アロエ多糖体が、グルコース、マンノース、アラビノース及びガラクトースからなる群から選択される単糖をさらに含む、請求項25に記載の組成物。
- アロエ多糖体が、11,500ダルトン〜10,000,000ダルトン超の範囲の分子量を有する、請求項25に記載の組成物。
- ポリマンナン抽出物が少なくとも25%のアロエ多糖体を有する、請求項21に記載の組成物。
- ポリマンナン抽出物が、25%、30%、40%、50%、60%、70%、80%、90%又は95%のアロエ多糖体を有する、請求項21に記載の組成物。
- ポリマンナン抽出物が、66,000ダルトンの分子量を有するアロエ多糖体を少なくとも14%含む、請求項21に記載の組成物。
- ポリマンナン抽出物が、480,000ダルトンの分子量を有するアロエ多糖体を少なくとも9%含む、請求項21に記載の組成物。
- ポリマンナン抽出物が、1,000,000ダルトンの分子量を有するアロエ多糖体を少なくとも3.5%含む、請求項21に記載の組成物。
- ポリマンナン抽出物が、2,000,000ダルトンの分子量を有するアロエ多糖体を少なくとも2.4%含む、請求項21に記載の組成物。
- 組成物が、白血病及びリンパ腫、前立腺がん、乳がん並びに結腸がんからなる群から選択される1又は2以上のがんの治療において使用される、請求項21に記載の組成物。
- 注射用製剤が、免疫調節、免疫刺激のために、又は免疫系の不全若しくは免疫疾患を有する個体の治療のために使用される、請求項21に記載の組成物。
- 1又は2以上のナチュラルキラー(NK)細胞の75〜80%の増加を引き起こす、請求項21に記載の組成物。
- 白血病及びリンパ腫、前立腺がん、乳がん並びに結腸がんからなる群から選択される1又は2以上のがんの治療のための方法であって、
1又は2以上のがんに対する治療を必要とする個体を特定するステップと、
無菌注射用ポリマンナン抽出物製剤を、週に2〜3回、前記1又は2以上のがんを治療するために十分な投薬量で注射するステップであって、前記無菌注射用ポリマンナン抽出物製剤が、脱イオン水に溶解された所定量の微細なポリマンナン抽出物及び1又は2以上の医薬品保存剤を含むステップと
を含む、前記方法。 - 血液試料を個体から1又は2以上の所定の間隔で抜き取るステップと、
前記血液中のカスパーゼ3タンパク質のレベルを測定し、取得した前記レベルを注射前のレベルと比較するステップであって、カスパーゼ3のレベル増加が、1又は2以上のがん細胞のアポトーシスのレベル増加と直接関係しているステップと
をさらに含む、請求項37に記載の方法。 - 無菌注射用ポリマンナン抽出物製剤の投薬量が、個体の体重、年齢、民族性及び性別によって決まる、請求項37に記載の方法。
- ポリマンナン抽出物がアロエ多糖体を含み、前記アロエ多糖体が、1又は2以上の短鎖、中鎖、長鎖、極長鎖多糖体、又はそれらの任意の組合せを含む、請求項37に記載の方法。
- アロエ多糖体が、11,500ダルトン〜10,000,000ダルトン超の範囲の分子量を有する、請求項40に記載の方法。
- ポリマンナン抽出物が少なくとも25%のアロエ多糖体を有する、請求項37に記載の方法。
- ポリマンナン抽出物が、1又は2以上のナチュラルキラー(NK)細胞の75〜80%の増加を引き起こす、請求項37に記載の方法。
- 免疫系の不全又は免疫疾患を有する個体における免疫調節又は免疫刺激の方法であって、
免疫系の不全又は免疫疾患を有し、免疫調節又は免疫刺激を必要とする個体を特定するステップと、
所定投薬量の無菌注射用ポリマンナン抽出物製剤を静脈内に投与するステップであって、前記無菌注射用ポリマンナン抽出物製剤が、脱イオン水に溶解された所定量の微細なポリマンナン抽出物及び1又は2以上の医薬品保存剤を含み、前記無菌注射用ポリマンナン抽出物製剤の投薬量が、前記個体の体重、年齢、民族性及び性別によって決まるステップと、
血液試料を前記個体から1又は2以上の所定の間隔で抜き取るステップと、
前記血液中の腫瘍壊死因子アルファ(TNFα)のレベルを測定し、取得された前記レベルを注射前のレベルと比較するステップであって、前記TNFαのレベル増加が免疫調節又は免疫刺激を示すステップと
を含む、前記方法。 - 免疫疾患が免疫に関連する腫瘍である、請求項44に記載の方法。
- ポリマンナン抽出物がアロエ多糖体を含み、前記アロエ多糖体が、1又は2以上の短鎖、中鎖、長鎖、極長鎖多糖体、又はそれらの任意の組合せを含む、請求項44に記載の方法。
- ポリマンナン抽出物が、1又は2以上のナチュラルキラー(NK)細胞の75〜80%の増加を引き起こす、請求項44に記載の方法。
- ポリマンナン抽出物が少なくとも25%のアロエ多糖体を有する、請求項44に記載の方法。
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