JP6790189B2 - Hsv−2のためのワクチン - Google Patents
Hsv−2のためのワクチン Download PDFInfo
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Description
本出願は、米国特許法119条(e)に基づき、2012年5月16日に出願された米国仮特許出願第61/647,764号、2012年8月3日に出願された同第61/679,387号、および2012年10月15日に出願された同第61/714,158号の利益を主張するものであり、それらは全て参照によりその全体が本明細書に組み込まれる。
配列表への参照
(技術分野)
(項目1)
(a)配列番号4のアミノ酸1〜450のうちの少なくとも75%が欠けており、また配列番号4の1055〜1374のアミノ酸のうちの少なくとも75%が欠けている、UL19ポリペプチドの免疫原性断片、
(b)配列番号12に記載される配列、
(c)少なくとも15個の隣接アミノ酸にわたり少なくとも85%のアミノ酸同一性を保持する、(a)または(b)の免疫原性変異体、
(d)(a)または(b)の免疫原性断片、および
(e)(a)、(b)、(c)、または(d)のキメラ融合物から成る群から選択される、HSV−2ポリペプチドの免疫原性断片。
(項目2)
項目1に記載のポリペプチドをコードする、単離されたポリヌクレオチド。
(項目3)
免疫原性薬学的組成物であって、
(i)
(a)配列番号4のアミノ酸1〜450のうちの少なくとも75%が欠けており、また配列番号4の1055〜1374のアミノ酸のうちの少なくとも75%が欠けている、UL19ポリペプチドの免疫原性断片、
(b)配列番号12に記載される配列、
(c)少なくとも15個の隣接アミノ酸にわたり少なくとも85%のアミノ酸同一性を保持する、(a)または(b)の免疫原性変異体、
(d)(a)または(b)の免疫原性断片、および
(e)(a)、(b)、または(c)のキメラ融合物から成る群から選択される、HSV−2ポリペプチドの免疫原性断片と、
(ii) 任意に、先天性免疫を活性化する薬剤と、
(iii) 薬学的に許容される担体と、を含む、組成物。
(項目4)
UL25またはその免疫原性断片をさらに含む、項目3に記載の組成物。
(項目5)
gD2またはその免疫原性断片をさらに含む、項目3および4のいずれか1項に記載の組成物。
(項目6)
前記薬剤は、アジュバントである、項目3〜5のいずれかに記載の組成物。
(項目7)
前記アジュバントは、GLAである、項目6に記載の組成物。
(項目8)
前記GLAは、水中油型エマルジョンの形態であるか、または水性形態である、項目7に記載の組成物。
(項目9)
前記水中油型エマルジョンは、スクアレンを含む、項目8に記載の組成物。
(項目10)
対象においてHSV−2感染を治療するための方法であって、前記対象に、項目3〜9のいずれか1項に記載の組成物を投与することを含む、方法。
(項目11)
対象において免疫応答を生じさせる方法であって、前記対象に、項目3〜9のいずれか1項に記載の組成物を投与することを含む、方法。
(項目12)
対象に、HSV−2に対して免疫付与するための方法であって、前記対象に、項目3〜9のいずれか1項に記載の組成物を投与することを含む、方法。
(項目13)
前記投与経路が、皮内、粘膜、筋肉内、皮下、舌下、直腸内、または膣内である、項目9〜12のいずれか1項に記載の方法。
(項目14)
前記対象に、項目3〜9のいずれか1項に記載の第2、第3、または第4の組成物を投与することをさらに含む、項目9〜13のいずれかに記載の方法。
本発明のこれらのおよび他の態様および実施形態は、以下の詳細な説明および添付の図面への参照により、明らかになるであろう。
A. ワクチンの構成要素としてのHSV−2タンパク質
et al. J Clin Invest 104:R63−R67, 1999;Altman et al., Science 274:94−96, 1996)が、とりわけ好適な分析である。場合により、断片(単数または複数)は、免疫優勢ペプチド配列を含む。幾つかの免疫優勢エピトープが、HSV−2糖タンパク質および構造タンパク質に関して同定されている(例えば、Kim et al. J Immunol 181:6604−6615, 2008;Chentoufi et al., J Virol.82:11792−11802, 2008;Koelle et al., Proc Natl Acad Sci USA 100:12899−12904, 2003、全ての参考文献はその全体が本明細書に組み込まれる)。 免疫原性ペプチドはまた、生物情報学ソフトウエアによって予測され得る(Flower, Methods in Molecular Biology vol.409, 2007)幾つかの例示的なプログラムおよびデータベースとしては、FRED(Feldhahn et al. Bioinformatics 15:2758−9, 2009)、SVMHC(Donnes and Kohlbacher,Nucleic Acids Res 34:W1940197,2006)、AntigenDB(Ansari et al., Nucleic Acids Res 38:D847−853,2010)、TEPITOPE(Bian and Hammer Methods 34:468−475, 2004)が挙げられる。
B. 先天性免疫を活性化する薬剤
C. 薬学的組成物および使用
1. 製剤
2. 投与
組換え発現ベクター、ウイルスベクター、およびウイルス様粒子
A. タンパク質の組換え産生
B. 対象へのタンパク質の送達のための組換え発現ベクター
(1993)を参照)。
C. ウイルスベクター
Tartaglia et al., AIDS Research and Human Retroviruses 8:1445−47 (1992)を参照)、またはMVA(例えば、Gheradi et al., J. Gen.Virol.86:2925−36 (2005); Mayr et al., Infection 3:6−14 (1975)を参照)を含む。また、Hu et al.(癌治療用のベクターとしてのヤバ様疾患ウイルスの使用を説明する、J.Virol.75:10300−308 (2001))、米国特許第5,698,530号および第6,998,252号も参照されたい。また、例えば、米国特許第5,443,964号も参照されたい。また、米国特許第7,247,615号および第7,368,116号も参照されたい。
D. レンチウイルスベクター
E. ウイルス様粒子
調節発現配列
and Taira, Human Gene Therapy, 11:577−585 (2000)およびMeissner et al., Nucleic Acids Res., 29:1672−1682 (2001)に提供されるPol IIまたはIIIプロモーターとしては、テトラサイクリン応答プロモーターが挙げられる。
al. 2004 Nat. Biotechnol. 22: 589−594を参照)に由来する2A様配列と結合されたフーリン開裂部位配列(RAKR)(例えば、Fang et al. 2005 Nat. Biotech. 23: 584−590を参照)等のオリゴヌクレオチドが、マルチシストロン性ベクター内の遺伝要素を分離するために使用される。特定のマルチシストロン性ベクターの有効性は、標準プロトコルを用いて遺伝子の各々の発現を検出することによって、容易に試験され得る。
マウスにおいて、複数のワクチン接種後にアジュバントGLA−SEとともに処方したときの、HSV−2GD2タンパク質に対するCD4T細胞ベースの免疫原性の強化
実施例2
GLAは、マウスにおいてCD8T細胞応答を増大させる
実施例3
マウスにおける複数のワクチン接種後の、個々のHSV−2 GD2、UL19、およびUL25タンパク質に対するCD4T細胞ベースの免疫原性
実施例4
マウスにおける、三価製剤の複数のワクチン接種の後の、それぞれの個々のHSV−2サブユニットタンパク質に対するCD4T細胞およびB細胞ベースの免疫原性
実施例5
マウスにおけるGLA−SEを伴うHSV−2UL19の単一および複数の免疫付与の後の、抗原特異的CD4T細胞応答
実施例6
マウスにおけるGLA−SEとともに処方される三価HSVワクチンを用いた免疫付与の後の、抗原特異的CD4T細胞応答
実施例7
マウスにおいて、複数のワクチン接種後にアジュバントGLA−SEとともに処方したときの、HSV−2GD2タンパク質に対する抗体ベースの免疫原性の強化
実施例8
アジュバントGLA−SEとともに処方したときの、HSV−2 UL19UDタンパク質に対するCD8T細胞ベースの免疫原性の強化
実施例9
アジュバントGLA−SEとともに処方されるときの、組換えHSV−2タンパク質ワクチンの予防的抗ウイルス有効性の強化
実施例10
アジュバントGLA−SEとともに処方されるときの、組換えHSV−2タンパク質ワクチンによる既存の記憶CD8T細胞の拡大の強化
実施例11
モルモットにおける再発性HSV−2を治療する組換えHSV−2タンパク質ワクチンの能力
実施例12
HSV−2エンベロープ糖タンパク質に由来し、リーダー配列を含有する、免疫原性タンパク質の構築
実施例13
有毒なHSV−2による致死的攻撃に対するGLA/SEおよび組換え三価タンパク質ワクチンの防御的有効性
10匹のC57BL/6マウスの群に、5μgのGLA−SEかまたは5%デキストロース媒介物かのいずれかと組み合わせて、各々5μgの組換えgD2、UL19ud(配列番号12参照)、およびUL25から成る三価ワクチンの2回の筋肉内免疫付与を、28日空けて行った。5μgのGLA−SE単独で免疫付与したマウスは、陰性対照の役割を果たした。追加の対照群は、5μgのGLA−SE、および飲料水中の1ml当たり1ミリグラムのアシクロビル(ACV)を用いて、攻撃の24時間後に開始して免疫付与されたマウスで構成された。第2の免疫付与の22日後、マウスは、デポ型酢酸メドロキシプロゲステロンで処理され、次いで6日後に、50xLD50用量の野生型HSV−2で膣内に攻撃された。マウスは、生殖器損傷形成および生存に関して毎日モニタリングされた。感染後第1日目、第3日目、および第5日目に、膣内スワブをPCRによるHSV−2DNAの定量化のために収集した。
図15に示されるように、三価組換えgD2、UL19ud、およびUL25を用いてGLA−SEとともに免疫付与したマウスは、三価タンパク質ワクチン単独かまたはGLA−SE単独かのいずれかを用いて免疫付与したマウスと比較して、劇的に低減された損傷形成(パネルA)を有し、また増加された生存率(パネルB)を有する。同様に、図16に示されるように、GLA−SEとともにgD2/UL19ud/UL25を用いて免疫付与した10匹のマウスのうち7匹は、第5日までに検出可能な膣内HSV−2DNAを有さず、一方、3つの全ての対照群のマウスは、第5日までを通して膣内に持続したレベルのHSV−2を有した。アシクロビルを受けた動物もまた、第1日、第3日、および第5日に、同程度に高いHSV−2DNAウイルス負荷を有した。GLA/SEおよびgD2/UL19ud/UL25の活性ワクチンを受けた動物は、特により低いウイルス負荷を有し、第5日までに多くの動物が無菌であった(すなわち、検出可能なウイルス負荷がなかった)。要約すると、これらの実験は、GLA/SE+組換え三価gD2/UL19ud/UL25タンパク質ワクチンの、有毒なHSV−2による致死的攻撃に対する生体内の防御的有効性を実証する。
実施例14
ヒトにおけるワクチンの安全性および免疫原性
Claims (10)
- 免疫原性薬学的組成物であって、
(a)(i) 配列番号4のアミノ酸1〜450のうちの少なくとも75%が欠けており、配列番号4の1055〜1374のアミノ酸のうちの少なくとも75%が欠けている、UL19ポリペプチドの免疫原性断片であるHSV−2ポリペプチドの免疫原性断片、または前記免疫原性断片の配列の全長にわたり少なくとも85%のアミノ酸同一性を保持する、その免疫原性変異体;
(ii)HSV−2 UL25タンパク質またはその免疫原性断片;および
(iii)HSV−2 gD2タンパク質またはその免疫原性断片と、
(b) モノホスホリル脂質Aアジュバントと、
(c) 薬学的に許容される担体と、を含み、
ここで、UL19ポリペプチドの免疫原性断片は、異種ペプチドに融合されている、免疫原性薬学的組成物。 - 異種ペプチドが、タグまたはマーカー配列である、請求項1に記載の免疫原性薬学的組成物。
- タグが、開裂配列を含んでいてもよいヒスチジンタグである、請求項2に記載の免疫原性薬学的組成物。
- 免疫原性薬学的組成物であって、
(a)(i) 配列番号4のアミノ酸1〜450のうちの少なくとも75%が欠けており、配列番号4の1055〜1374のアミノ酸のうちの少なくとも75%が欠けている、UL19ポリペプチドの免疫原性断片であるHSV−2ポリペプチドの免疫原性断片、または前記免疫原性断片の配列の全長にわたり少なくとも85%のアミノ酸同一性を保持する、その免疫原性変異体;
(ii)HSV−2 UL25タンパク質またはその免疫原性断片;および
(iii)HSV−2 gD2タンパク質またはその免疫原性断片と、
(b) モノホスホリル脂質Aアジュバントと、
(c) 薬学的に許容される担体と、を含み、
ここで、gD2タンパク質またはその免疫原性断片、および/または、UL25タンパク質またはその免疫原性断片が、異種ペプチドに融合されている、免疫原性薬学的組成物。 - 前記モノホスホリル脂質Aアジュバントは、GLAである、請求項1〜4のいずれか1項に記載の組成物。
- 前記GLAは、水中油型エマルジョンの形態であるか、または水性形態である、請求項5に記載の組成物。
- 前記水中油型エマルジョンは、スクアレンを含む、請求項6に記載の組成物。
- 対象においてHSV−2感染を治療するための請求項1〜7のいずれか1項に記載の組成物。
- 対象においてHSV−2に対する免疫応答を生じさせるための請求項1〜7のいずれか1項に記載の組成物。
- 皮内、粘膜、筋肉内、皮下、舌下、直腸内、または膣内投与されることを特徴とする、請求項1〜9のいずれか1項に記載の組成物。
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