JP2018516926A - ブタ生殖器呼吸器症候群及びブタサーコウイルス関連疾患に対するワクチン組成物 - Google Patents
ブタ生殖器呼吸器症候群及びブタサーコウイルス関連疾患に対するワクチン組成物 Download PDFInfo
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Abstract
Description
本発明は、一般にワクチンに関し、より具体的にはサブユニットワクチンに関する。
免疫細胞(T細胞、B細胞、樹状細胞、単球、又はマクロファージなど)に感染するウイルスとしては、ブタ生殖器呼吸器症候群ウイルス(PRRSV)、ブタサーコウイルスタイプII(PCV2)、及びヒト免疫不全ウイルス(HIV)が含まれる。免疫細胞は、免疫化応答を誘発することはできないが、ウイルスを運ぶことができる。これらのウイルスに感染した動物には、他の病原体が容易に感染する可能性がある。ブタ生殖器呼吸器症候群ウイルス(PRRSV)は、毎年、畜産に大きな損失をもたらす。ブタだけでなく、アヒルも同様に、PRRSVに感染する可能性がある。一般に、ウイルスに感染した動物は、顕著な症状を有しないが、感染した動物の免疫性は低下する。このウイルスは、(肺胞及び脾臓中の)マクロファージ、脳ミクログリア及び単球に感染し、感染した動物の血液及び器官に存在し得る。これは、二次感染に起因する体重増加の減少及び死亡率の増加につながる。
一つの態様において、本発明は、ブタ生殖器呼吸器症候群ウイルス(PRRSV)融合タンパク質に関する。このブタ生殖器呼吸器症候群ウイルス(PRRSV)融合タンパク質は、
(a)融合タンパク質のN末端に位置する、抗原提示細胞(APC)結合ドメイン又はCD91レセプター結合ドメイン;
(b)APC結合ドメイン又はCD91レセプター結合ドメインのC末端に位置する、配列番号:4、2、3、又は6と少なくとも90%同一のアミノ酸配列を含む、34−112アミノ酸残基長のトランスロケーションペプチド;
(c)以下の(i)〜(iv)を含む融合抗原;
(i)ブタ生殖器呼吸器症候群ウイルス(PRRSV)ORF7抗原;
(ii)PRRSV ORF1b抗原;
(iii)PRRSV ORF6抗原;及び
(iv)PRRSV ORF5抗原;
(d)融合タンパク質のC末端に位置する、小胞体保持配列;及び
(e)任意選択的に、抗原と小胞体保持配列との間又はトランスロケーションペプチドと抗原との間に位置する、配列番号:13のアミノ酸配列を含む核輸送シグナル
を含み、
ここで、融合抗原が、完全長ORF7、ORF6、ORF5、及びORF1bタンパク質配列を含まない。
(i)本発明のPRRSV融合タンパク質;及び
(ii)ブタサーコウイルスタイプ2(PCV2)融合タンパク質
を含み、
(ii)ブタサーコウイルスタイプ2(PCV2)融合タンパク質が、
(a)融合タンパク質のN末端に位置する、抗原提示細胞(APC)結合ドメイン又はCD91レセプター結合ドメイン;
(b)APC結合ドメイン又はCD91レセプター結合ドメインのC末端に位置する、配列番号:4、2、3、又は6と少なくとも90%同一のアミノ酸配列を含む、34−112アミノ酸残基長のトランスロケーションペプチド;及び
(c)PCV2 ORF2抗原;
(d)融合タンパク質のC末端に位置する、小胞体保持配列;及び
(e)抗原と小胞体保持配列との間又はトランスロケーションペプチドと抗原との間に位置する、配列番号:13のアミノ酸配列を含む核輸送シグナル
を含み、
ここで、PCV2 ORF2抗原が、PCV2 ORF2タンパク質のC末端部アミノ酸配列を含み、PCV2融合タンパク質が、PCV2 ORF2タンパク質のN末端部アミノ酸配列を含まない。
本発明は、以下の実施例においてより具体的に説明されている。これらの実施例は、単なる例示であることが意図される。なぜなら、実施例における多数の改変及び変更が当業者には明らかであるためである。本発明の様々な実施形態をここで詳細に説明する。図面を参照すると、図面全体において、類似の番号は類似の構成要素を示す。この詳細な説明において、及び以下に続く特許請求の範囲全体において使用されるように、「a」、「an」、及び「the」の意味は、文脈上他に明確に指示されない限り、複数の参照を含む。また、この詳細な説明において、及び以下に続く特許請求の範囲全体において使用されるように、「in」の意味は、文脈上他に明確に指示されない限り、「in」及び「on」を含む。さらに、読者の便宜のために、タイトル又はサブタイトルが明細書中で使用され得る。このことは、本発明の範囲に影響を与えるものではない。加えて、この明細書において使用されるいくつかの用語が、以下により具体的に定義される。
(i)PRRSV ORF7のC末端部の2回の反復を含む抗原をコードする標的DNA。この抗原は、「DGD」又は「D」と呼ばれる。
(ii)PRRSV NSP 10(C末端ドメイン配列)及びNSP 11(N末端ドメイン配列)を融合して得られる融合抗原をコードする標的DNA。この抗原は、「M12」又は「M」と呼ばれる。
(iii)ORF6配列とORF5配列との間に架橋/リンカー配列を有しない、PRRSV ORF6のN末端部及びORF5のN末端部を融合して得られる融合抗原をコードする標的DNA。この抗原は、「RSAB」又は「R」と呼ばれる。
(iv)ORF6配列とORF5配列との間に架橋/リンカー配列を有しない、PRRSV ORF6のN末端部及びORF5のN末端部を融合して得られる融合抗原をコードする標的DNA。この抗原は、「PQAB」又は「P」と呼ばれる。
**:ボールド体の文字は、人工核輸送シグナルのアミノ酸配列を表す;下線の付された文字は、小胞体保持シグナルのアミノ酸配列を表す。
***:M(M12)は、PRRSV NSP 10(C末端ドメイン配列)及びNSP 11(N末端ドメイン配列)の融合によって調製された融合ポリペプチドである。すなわち、このポリペプチドは、非構造タンパク質であるORF1b NSP 10 C末端部及びNSP 11 N末端部に由来する。
****:P(PQAB)は、PRRSV ORF6 a.a.2−a.a.26及びORF5 aa31−aa63の融合によって調製されたポリペプチドである。米国特許第7465455号を参照。普通の文字で示される配列はPRRSV ORF6/マトリクスタンパク質配列に由来し、ボールド体の文字で示される配列はPRRSV ORF5配列に由来する。PRRSVのORF5によってコードされる主要なエンベロープタンパク質(GP5)は、ウイルス中和(VN)抗体及びPRRSVの異なる株間の交差防御の誘導において、決定的役割を有する。異なる株間には配列変異があるため、本明細書中の配列は、例示の目的で開示される。
Claims (15)
- 融合タンパク質であって、
(a)前記融合タンパク質のN末端に位置する、抗原提示細胞(APC)結合ドメイン又はCD91レセプター結合ドメイン;
(b)前記APC結合ドメイン又は前記CD91レセプター結合ドメインのC末端に位置する、配列番号:4、2、3、又は6と少なくとも90%同一のアミノ酸配列を含む、34−112アミノ酸残基長のトランスロケーションペプチド;
(c)以下の(i)〜(iv)を含む融合抗原;
(i)ブタ生殖器呼吸器症候群ウイルス(PRRSV)ORF7抗原;
(ii)PRRSV ORF1b抗原;
(iii)PRRSV ORF6抗原;及び
(iv)PRRSV ORF5抗原;
(d)前記融合タンパク質のC末端に位置する、小胞体保持配列;及び
(e)任意選択的に、前記抗原と前記小胞体保持配列との間又は前記トランスロケーションペプチドと前記抗原との間に位置する、配列番号:13のアミノ酸配列を含む核輸送シグナル
を含み、
ここで、前記融合抗原が、完全長ORF7、ORF6、ORF5、及びORF1bタンパク質配列を含まない、
融合タンパク質。 - 前記ORF7又はORF1b抗原が前記ORF6抗原のN末端に位置し、前記ORF5抗原が前記ORF6抗原のC末端に位置する、請求項1に記載の融合タンパク質。
- 前記融合抗原が、前記ORF7抗原の2つのタンデム反復を含む、請求項1に記載の融合タンパク質。
- 前記ORF5抗原が前記ORF6抗原のC末端に位置する、請求項1に記載の融合タンパク質。
- 前記ORF6抗原が、前記PRRSV ORF6のN末端部アミノ酸配列を含み、前記ORF5抗原が、前記PRRSV ORF5のN末端部アミノ酸配列を含み、前記融合抗原が、前記ORF6及びORF5のC末端部アミノ酸配列を含まない、請求項1に記載の融合タンパク質。
- 前記ORF1b抗原が、ORF1b NSP 10のC末端部アミノ酸配列及びORF1b NSP 11のN末端部アミノ酸配列を含み、前記融合抗原が、前記ORF1bのN末端及びC末端部アミノ酸配列を欠いている、請求項5に記載の融合タンパク質。
- 前記APC結合ドメイン又は前記CD91レセプター結合ドメインが、配列番号:1、8、9、10、11、又は32と少なくとも90%同一のアミノ酸配列を含むポリペプチドである、請求項1に記載の融合タンパク質。
- 前記トランスロケーションペプチドが34−61アミノ酸残基長を有する、請求項1に記載の融合タンパク質。
- 前記小胞体保持配列がアミノ酸配列KDEL(配列番号:15)を含み、前記アミノ酸KDELのタンデム反復を含まず、但し、前記核輸送シグナルが存在する、請求項1に記載の融合タンパク質。
- 前記核輸送シグナル及び前記ER保持配列が、配列番号:12と少なくとも90%同一のアミノ酸配列と融合ペプチドを形成する、請求項9に記載の融合タンパク質。
- 前記小胞体保持配列が配列番号:16、17、18、又は19のアミノ酸配列を有し、但し、前記核輸送シグナルが存在しない、請求項1に記載の融合タンパク質。
- 前記APC結合ドメイン又は前記CD91レセプター結合ドメインが、配列番号:8と少なくとも90%同一のアミノ酸配列を含むポリペプチドである、請求項11に記載の融合タンパク質。
- 組成物であって、
(i)請求項1に記載の融合タンパク質;及び
(ii)ブタサーコウイルスタイプ2(PCV2)融合タンパク質を含み、
(ii)ブタサーコウイルスタイプ2(PCV2)融合タンパク質が、
(a)前記融合タンパク質のN末端に位置する、抗原提示細胞(APC)結合ドメイン又はCD91レセプター結合ドメイン;
(b)前記APC結合ドメイン又は前記CD91レセプター結合ドメインのC末端に位置する、配列番号:4、2、3、又は6と少なくとも90%同一のアミノ酸配列を含む、34−112アミノ酸残基長のトランスロケーションペプチド;及び
(c)PCV2 ORF2抗原;
(d)前記融合タンパク質のC末端に位置する、小胞体保持配列;及び
(e)前記抗原と前記小胞体保持配列との間又は前記トランスロケーションペプチドと前記抗原との間に位置する、配列番号:13のアミノ酸配列を含む核輸送シグナル
を含み、
ここで、前記PCV2 ORF2抗原が、PCV2 ORF2タンパク質のC末端部アミノ酸配列を含み、前記PCV2融合タンパク質が、前記PCV2 ORF2タンパク質のN末端部アミノ酸配列を含まない、
組成物。 - 前記トランスロケーションペプチドが34−61アミノ酸残基長を有する、請求項1に記載の融合タンパク質。
- 抗原特異的細胞媒介性応答及び体液性応答を、それを必要とする被験体において誘導するための医薬品の製造における、治療的に有効量の、請求項1に記載の融合タンパク質又は請求項13に記載の組成物を含む組成物の使用。
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KR102041032B1 (ko) | 2019-11-27 |
EP3302543A4 (en) | 2018-12-05 |
CA2985381A1 (en) | 2016-12-08 |
AU2016271144A1 (en) | 2017-11-30 |
JP6676661B2 (ja) | 2020-04-08 |
KR20180015185A (ko) | 2018-02-12 |
EP3302543B1 (en) | 2020-04-29 |
TW201706296A (zh) | 2017-02-16 |
WO2016196383A1 (en) | 2016-12-08 |
CN107847575B (zh) | 2021-12-17 |
TWI621627B (zh) | 2018-04-21 |
EP3302543A1 (en) | 2018-04-11 |
AU2016271144B2 (en) | 2018-10-25 |
US20160346377A1 (en) | 2016-12-01 |
US10010602B2 (en) | 2018-07-03 |
ES2790527T3 (es) | 2020-10-28 |
DK3302543T3 (da) | 2020-05-18 |
CA2985381C (en) | 2020-03-24 |
CN107847575A (zh) | 2018-03-27 |
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