JP2021513859A - ナチュラルキラー細胞の活性化および拡大のための方法ならびにその使用 - Google Patents
ナチュラルキラー細胞の活性化および拡大のための方法ならびにその使用 Download PDFInfo
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Abstract
Description
本願は、2018年2月21日出願の米国特許仮出願第62/633,592号の利益を主張し、その全体が参照により本明細書中に援用される。
I.定義
II.ナチュラルキラー(NK)細胞
A.キメラ抗原レセプター
B.抗原提示細胞
III.使用方法
IV.キット
以下の実施例は、本発明の好ましい実施形態を示すために含められる。以下の実施例に開示される手法は、本発明の実施においてうまく機能すると本発明者が発見した手法であり、ゆえに、それらの手法は、実施するための好ましい形態を構成すると見なされ得ることが、当業者によって認識されるべきである。しかしながら、当業者は、本開示に鑑みて、開示される具体的な実施形態では、本発明の趣旨および範囲から逸脱することなく多くの変更が行われ得、なおも同様または類似の結果が得られることを認識するはずである。
実施例1−NK細胞のプレ活性化および拡大
以下の参考文献は、それらが本明細書に記載されたものを補足する例示的な手順または他の詳細を提供する範囲で、参照により本明細書に具体的に援用される。
国際公開番号 WO2007/103009
Leong et al., Biol Blood Marrow Transplant. 20(4):463-73, 2014.
Singh et al., Cancer Research, 71:3516-3527, 2011.
U.S. Patent No. 4,690,915
U.S. Patent No. 6,225,042
U.S. Patent No. 6,355,479
U.S. Patent No. 6,362,001
U.S. Patent No. 6,410,319
U.S. Patent No. 6,790,662
U.S. Patent No. 7,109,304
U.S. Patent Publication No. 2009/0004142
U.S. Patent Publication No. 2009/0017000
Claims (75)
- ナチュラルキラー(NK)細胞を拡大するためのインビトロ方法であって、
(a)NK細胞の集団を得る工程;
(b)該NK細胞の集団を、有効濃度のIL−12、IL−15およびIL−18を含むプレ活性化培養物中でプレ活性化して、プレ活性化されたNK細胞を得る工程;および
(c)該プレ活性化されたNK細胞を、CD137リガンドを発現する人工抗原提示細胞(aAPC)を含む拡大培養物中で拡大することにより、拡大されたNK細胞を生成する工程
を含む、方法。 - 前記NK細胞の集団が、臍帯血(CB)、末梢血(PB)、幹細胞または骨髄から得られる、請求項1に記載の方法。
- 前記幹細胞が、誘導多能性幹細胞である、請求項2に記載の方法。
- 前記NK細胞の集団が、CBから得られる、請求項1に記載の方法。
- 前記CBが、2単位以上の個々の臍帯血単位からプールされる、請求項4に記載の方法。
- 前記CBが、3、4、5、6、7または8単位の個々の臍帯血単位からプールされる、請求項4に記載の方法。
- 前記NK細胞の集団が、CB単核細胞(CBMC)である、請求項1に記載の方法。
- 前記NK細胞の集団がさらに、CD56+NK細胞と定義される、請求項1に記載の方法。
- 前記aAPCがさらに、膜結合型サイトカインを発現する、請求項1に記載の方法。
- 前記膜結合型サイトカインが、膜結合型IL−21(mIL−21)または膜結合型IL−15(mIL−15)である、請求項9に記載の方法。
- 前記膜結合型サイトカインが、mIL−21である、請求項9に記載の方法。
- 前記aAPCが、内在性のHLAクラスI、IIまたはCD1d分子を本質的に発現しない、請求項1〜11のいずれか1項に記載の方法。
- 前記aAPCが、ICAM−1(CD54)およびLFA−3(CD58)を発現する、請求項1に記載の方法。
- 前記aAPCがさらに、白血病細胞由来aAPCと定義される、請求項1に記載の方法。
- 前記白血病細胞由来aAPCが、CD137リガンドおよび/またはmIL−21を発現するように操作されたK562細胞である、請求項14に記載の方法。
- 前記K562細胞が、CD137リガンドおよびmIL−21を発現するように操作される、請求項15に記載の方法。
- 前記aAPCが、レトロウイルス導入によって操作されたものである、請求項15に記載の方法。
- 前記aAPCが、照射される、請求項1に記載の方法。
- 前記プレ活性化工程が、10〜20時間である、請求項1に記載の方法。
- 前記プレ活性化工程が、14〜18時間である、請求項1に記載の方法。
- 前記プレ活性化工程が、16時間である、請求項1〜20のいずれか1項に記載の方法。
- 前記プレ活性化培養物が、IL−18および/またはIL−15を10〜100ng/mLの濃度で含む、請求項1に記載の方法。
- 前記プレ活性化培養物が、IL−18および/またはIL−15を40〜60ng/mLの濃度で含む、請求項1に記載の方法。
- 前記プレ活性化培養物が、IL−18および/またはIl−15を50ng/mLの濃度で含む、請求項1に記載の方法。
- 前記プレ活性化培養物が、IL−12を0.1〜150ng/mLの濃度で含む、請求項1に記載の方法。
- 前記プレ活性化培養物が、IL−12を1〜20ng/mLの濃度で含む、請求項1に記載の方法。
- 前記プレ活性化培養物が、IL−12を10ng/mLの濃度で含む、請求項1に記載の方法。
- 前記プレ活性化されたNK細胞を拡大前に洗浄する工程をさらに含む、請求項1に記載の方法。
- 洗浄が、複数回行われる、請求項28に記載の方法。
- 拡大が、5〜20日間である、請求項1に記載の方法。
- 前記拡大が、12〜16日間である、請求項1に記載の方法。
- 前記拡大が、14日間である、請求項1に記載の方法。
- 前記プレ活性化されたNK細胞とaAPCとが、前記拡大培養物中に3:1〜1:3の比で存在する、請求項1に記載の方法。
- 前記プレ活性化されたNK細胞とaAPCとが、前記拡大培養物中に1:2の比で存在する、請求項1に記載の方法。
- 前記拡大培養物が、IL−2をさらに含む、請求項1〜34のいずれか1項に記載の方法。
- 前記IL−2が、10〜500U/mLの濃度で存在する、請求項35に記載の方法。
- 前記IL−2が、100〜300U/mLの濃度で存在する、請求項35に記載の方法。
- 前記IL−2が、200U/mLの濃度で存在する、請求項35に記載の方法。
- 前記IL−12、IL−18、IL−15および/またはIL−2が、組換えヒトIL−2である、請求項35に記載の方法。
- 前記IL−2が、前記拡大培養物に2〜3日ごとに補充される、請求項35に記載の方法。
- 前記aAPCが、少なくとも2度、前記拡大培養物に加えられる、請求項1に記載の方法。
- 前記方法が、無血清培地中で行われる、請求項1に記載の方法。
- 前記NK細胞が、キメラ抗原レセプター(CAR)を発現するように操作される、請求項1〜42のいずれか1項に記載の方法。
- 前記CARが、ヒト化抗原結合ドメインを含む、請求項43に記載の方法。
- 前記CARが、CD19、CD123、メソテリン、CD5、CD47、CLL−1、CD33、CD99、U5snRNP200、CD200、CS1、BAFF−R、ROR−1またはBCMAの抗原結合ドメインを含む、請求項43に記載の方法。
- 前記CARが、CD19またはCD123の抗原結合ドメインを含む、請求項43に記載の方法。
- 前記CARが、IL−15を含む、請求項43に記載の方法。
- 前記CARが、自殺遺伝子を含む、請求項43に記載の方法。
- 前記自殺遺伝子が、CD20、CD52、EGFRv3または誘導性カスパーゼ9である、請求項43に記載の方法。
- 請求項1〜49のいずれか1項に記載の方法に従って作製され、拡大されたNK細胞の集団。
- 請求項50に記載の拡大されたNK細胞の集団および薬学的に許容され得るキャリアを含む薬学的組成物。
- 被験体における疾患または障害の処置において使用するための、有効量の請求項50に記載の拡大されたNK細胞を含む組成物。
- 前記疾患または障害が、癌、炎症、移植片対宿主病、移植片拒絶、自己免疫障害、免疫不全症、B細胞悪性疾患または感染症である、請求項52に記載の組成物。
- 前記癌が、白血病である、請求項53に記載の組成物。
- 前記白血病が、急性リンパ芽球性白血病(ALL)、慢性リンパ性白血病(CLL)、急性骨髄性白血病(AML)または慢性骨髄性白血病(CML)である、請求項54に記載の組成物。
- 前記障害が、移植片対宿主病(GVHD)である、請求項52に記載の組成物。
- 前記障害が、多発性硬化症、炎症性腸疾患、関節リウマチ、I型糖尿病、全身性エリテマトーデス、接触過敏症、喘息またはシェーグレン症候群である、請求項52に記載の組成物。
- 被験体における疾患または障害を処置する方法であって、治療有効量の請求項50に記載の拡大されたNK細胞を該被験体に投与する工程を含む、方法。
- 前記疾患または障害が、癌、炎症、移植片対宿主病、移植片拒絶、自己免疫障害、免疫不全症、B細胞悪性疾患または感染症である、請求項58に記載の方法。
- 前記癌が、白血病である、請求項58に記載の方法。
- 前記白血病が、急性リンパ芽球性白血病(ALL)、慢性リンパ性白血病(CLL)、急性骨髄性白血病(AML)または慢性骨髄性白血病(CML)である、請求項60に記載の方法。
- 前記NK細胞が、同種異系である、請求項58に記載の方法。
- 前記NK細胞が、自家性である、請求項58に記載の方法。
- 前記障害が、移植片対宿主病(GVHD)である、請求項58に記載の方法。
- 前記障害が、多発性硬化症、炎症性腸疾患、関節リウマチ、I型糖尿病、全身性エリテマトーデス、接触過敏症、喘息またはシェーグレン症候群である、請求項58に記載の方法。
- 前記被験体が、ヒトである、請求項58に記載の方法。
- 少なくとも1つの第2の治療薬を投与する工程をさらに含む、請求項58〜66のいずれか1項に記載の方法。
- 前記少なくとも1つの第2の治療薬が、治療有効量の抗癌剤、免疫調節剤または免疫抑制剤である、請求項67に記載の方法。
- 前記抗癌剤が、化学療法、放射線療法、遺伝子療法、手術、ホルモン療法、抗血管新生療法または免疫療法である、請求項68に記載の方法。
- 前記免疫抑制剤が、カルシニューリン阻害剤、mTOR阻害剤、抗体、化学療法剤照射、ケモカイン、インターロイキン、またはケモカインもしくはインターロイキンの阻害剤である、請求項68に記載の方法。
- NK細胞および/または前記少なくとも1つの第2の治療薬が、静脈内に、腹腔内に、気管内に、腫瘍内に、筋肉内に、内視鏡的に、病巣内に、経皮的に、皮下に、領域性に、または直接注射もしくは灌流によって投与される、請求項67に記載の方法。
- 前記第2の治療薬が、抗体である、請求項67に記載の方法。
- 前記抗体が、モノクローナル抗体、二重特異性抗体または三重特異性抗体である、請求項72に記載の方法。
- 前記抗体が、モノクローナル抗体である、請求項72に記載の方法。
- 前記抗体が、リツキシマブである、請求項72に記載の方法。
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US20210077527A1 (en) * | 2019-09-13 | 2021-03-18 | The Research Institute At Nationwide Children's Hospital | Universal donor selection method to identify nk-cell-donors |
US20210260115A1 (en) * | 2020-02-21 | 2021-08-26 | FullHope Biomedical Co., Ltd. | Modified natural killer cells, pharmaceutical composition, manufacturing method thereof, and method of using the same |
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