TWI293957B - A superantigen fusion protein and the use thereof - Google Patents
A superantigen fusion protein and the use thereof Download PDFInfo
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- TWI293957B TWI293957B TW093121720A TW93121720A TWI293957B TW I293957 B TWI293957 B TW I293957B TW 093121720 A TW093121720 A TW 093121720A TW 93121720 A TW93121720 A TW 93121720A TW I293957 B TWI293957 B TW I293957B
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- Taiwan
- Prior art keywords
- peptide
- fusion protein
- acute respiratory
- respiratory syndrome
- severe acute
- Prior art date
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- COYHRQWNJDJCNA-NUJDXYNKSA-N Thr-Thr-Thr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O COYHRQWNJDJCNA-NUJDXYNKSA-N 0.000 description 1
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- HPYDSVWYXXKHRD-VIFPVBQESA-N Tyr-Gly Chemical compound [O-]C(=O)CNC(=O)[C@@H]([NH3+])CC1=CC=C(O)C=C1 HPYDSVWYXXKHRD-VIFPVBQESA-N 0.000 description 1
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- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
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- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
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Description
1293957 九、發明說明： 【發明所屬之技術領域】 本發明係關於一種超抗原融合蛋白及其應用方法，尤指 一種可接合於抗原呈現細胞，有效誘發抗体，以抑制病毒感 染以及阻斷超抗原反應之融合蛋白。 【先前技術】 病毒微小，無法獨立繁殖，必須寄生於細胞内方能生 存，所以屬於生物和非生物之間；冠狀病毒（Coronaviruses) 是一單股RNA病毒，和目標細胞上的接受體結合後，形成一 囊泡，經由内吞作用（endocytosis )進入細胞内，藉由反轉 錄，能將病毒的核糖核酸（RNA )轉為DN A，再嵌入細胞染 色體中，利用宿主細胞製造病毒所需的蛋白及遺傳物質，在 酵素的催化下，蛋白和遺傳物質組裝後，從細胞内釋放出， 並破壞宿主細胞。 病毒認定細胞表面上接受體，例如HIV認定T細胞表面 上的CD4和CCR5抗原後，方能進入細胞；而冠狀病毒則認 定肺臟或腎臟等細胞上的胺基（Aminopeptidase-N )，即CD13 接受器之結構方能入侵。因此以生物科技發展對抗病毒的重 要方向，就是設計藥物阻止病毒和表面抗原結合，以阻止病 毒入侵；但是上述病菌具備超抗原，可直接與T細胞上接受 器結合後誘發T細胞產生大量之細胞素（interlukin)或 γ-interferon而導致激烈發炎反應，甚而讓結合之T細胞進入 1293957 死亡路徑（如程式死亡apoptosis)，因此如果能夠去除超抗原 與細胞之結合機制，又可阻斷病毒由細胞接受器侵入，則有 可能防止或減輕感染症狀。 在免疫系統中，每個T細胞的細胞膜上都會表現屬於自 己的T細胞接受體（T Cell Receptor，TCR)，大約有一百萬個 成熟的T細胞在我們的身體中巡邏，透過TCR來偵測身體内 體細胞或者抗原呈現之細胞膜上所帶有的訊息，抗原呈現細 胞會有一種可供辨識外來蛋白質之複合體Major Histocomparibility Complex (MHC)存在，在 MHC與一段胜肽 (peptide)結合後才呈現於細胞膜表面，而MHC與胜肽所形成 的複合體，就是專門來提供訊息給TCR，作為辨識自我與非 我的媒介；然而，經由科學家的致力研究，發現SARS病毒 在侵入體内後，其外套膜上之棘蛋白（spike protein)某一 部份會與T細胞上之TCR結合，且在不需要抗原呈現細胞的 MHC分子的參與，而立即釋出誤認為該正常細胞係一外來侵 入者之訊息，因而誘發T細胞的大量增殖或產生大量細胞 素，因而大肆攻擊自身細胞，引發免疫或發炎反應；根據以 上研究，可確知SARS棘蛋白確實有「超抗原（super antigen)」 之特性，但是此段超抗原之胺基酸序列座落於何部分，尚須 詳實探討，而近來有研究推測該超抗原之位置係座落於 SARS棘蛋白胺基酸序列第680至1050個位置，但仍須進一步 證實。 要預防SARS病毒入侵時引發自體免疫反應，可藉由類 似疫苗的系統達成。而接種疫苗的關鍵在於使最終被感染的 1293957 序列表 <110〉 <120> <130〉 <160〉 <170〉 <210> <211〉 <212> <213〉 <300〉 <308〉 <309〉 <313〉 <400〉 生寶臍帶血銀行 超抗原融合蛋白及其應用方法 P7133/0357 8
Patentln version 3.3 1 71
PRT 嚴重急性呼吸道症候群病毒E2棘蛋白 NP一828851 2004-04-09 ⑴"(71) 1
Asn Thr lie Ala lie Pro Thr Asn Phe Ser lie Ser lie Thr.Thr Glu 15 10 15
Val Met Pro Val Ser Met Ala Lys Thr Ser Val Asp Cys Asn Met Tyr 20 25 30 lie Cys Gly Asp Ser Thr Glu Cys Ala Asn Leu Leu Leu Gin Tyr Gly 22 1293957 35 40 45
Ser Phe Cys Thr Gin Leu Asn Arg Ala Leu Ser Gly lie Ala Ala Glu 50 55 60
Gin Asp Arg Asn Thr Arg Glu 65 70 <210〉 2 <211〉 97
<212> PRT <213〉 嚴重急性呼吸道症候群病毒之E2棘蛋白 <300〉 <308> NP_828851 <309〉 2004-04 - 09 <313〉 (1)..(97)
<400〉 2
Val Phe Ala Gin Val Lys Gin Met Tyr Lys Thr Pro Thr Leu Lys Tyr 1 5 10 15
Phe Gly Gly Phe Asn Phe Ser Gin lie Leu Pro Asp Pro Leu Lys Pro 20 25 23 1293957 30
Thr Lys Arg Ser Phe lie Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 35 40 45
Ala Asp Ala Gly Phe Met Lys Gin Tyr Gly Glu Cys Leu Gly Asp lie 50 55 60
Asn Ala Arg Asp Leu lie Cys Ala Gin Lys Phe Asn Gly Leu Thr Val 65 70 75 80
Leu Pro Pro Leu Leu Thr Asp Asp Met lie Ala Ala Tyr Thr Ala Ala 85 90 95
Leu <210> 3 <211> 90
<212> PRT <213> 嚴重急性呼吸道症候群病毒之E2棘蛋白 24 ， 1293957 <300> <308> NP__828851 <309〉 2004-04-09 <313〉 ⑴"(90) <400> 3
Val Ser Gly Thr Ala Thr Ala Gly Trp Thr Phe Gly Ala Gly Ala Ala 1 5 10 15
Leu Gin lie Pro Phe Ala Met Gin Met Ala Tyr Arg Phe Asn Gly lie
Gly Val Thr Gin Asn Val Leu Tyr Glu Asn Gin Lys Gin lie Ala Asn 35 40 45
Gin Phe Asn Lys Ala lie Ser Gin lie Gin Glu Ser Leu Thr Thr Thr 50 55 60
Ser Thr Ala Leu Gly Lys Leu Gin Asp Val Val Asn Gin Asn Ala Gin 65 70 75 25 1293957 80
Ala Leu Asn Thr Leu Val Lys Gin Leu Ser 85 90 <210> 4
<211> 101 <212> PRT
<213> 嚴重急性呼吸道症候群病毒之E2棘蛋白 <300> <308> NP—828851 <309> 2004-04-09 <313> (1)..(101) <400> 4
Ser Asn Phe Gly Ala lie Ser Ser Val Leu Asn Asp lie Leu Ser Arg 1 5 10 15
Leu Asp Lys Val Glu Ala Glu Val Gin lie Asp Arg Len lie Thr Gly
Arg Leu Gin Ser Leu Gin Thr Tyr Val Thr Gin Gin Leu lie Arg Ala 35 40 26 45 1293957
Ala Glu lie Arg Ala Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu 55 50 60
Cys Val Leu Gly Gin Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr 65 70 75 80
His Leu Met Ser Phe Pro Gin Ala Ala Pro His Gly Val Val Phe Leu 85 90 95
His Val Thr Tyr Val 100 <210> 5 <211〉 219 <212> DNA <213〉 人工合成 <220〉 <223〉 編碼有可於大腸桿菌表現系統中表現出嚴重急性 呼吸道症候群病毒E2棘蛋白之核酸序列 <400> 5 27 1293957 aacaccatcg ctatcccgac caacttctcc atctccatca ccaccgaagt tatgccggtt 60 tccatggcta aaacctccgt tgactgcaac atgtacatct gcggtgactc caccgaatgc 120 gctaacctgc tgctgcagta cggttccttc tgcacccagc tgaaccgtgc tctgtccggt 180 atcgctgctg aacaggaccg taacacccgt gaataatag 219 <210〉 6 <211〉 297 <212> DNA <213> 人工合成 <220〉 <223〉 編碼有可於大腸桿菌表現系統中表現出嚴重急性 呼吸道症候群病毒E 2棘蛋白之核酸序列 <400〉 6 gttttcgctc aggttaaaca gatgtacaaa accccgaccc tgaaatactt cggtggtttc 60 aacttctccc agatcctgcc ggacccgctg aaaccgacca aacgttcctt catcgaagac 120 1293957 ctgctgttca cggtgaatgc ctgggtgaca tctgaccgtt ctgccgccgc gtaatag acaaagttac 180 tcaacgctcg 240 tgctgaccga 297 cctggctgac tgacctgatc cgacatgatc gctggtttca tgaaacagta tgcgctcaga aattcaacgg gctgcttaca ccgctgctct <210> 7 <211> 276 <212〉 DNA <213〉 人工合成 <220> <223> 編碼有可於大腸桿菌表現系統中表現出嚴重急性呼 吸道症候群病毒E2棘蛋白之核酸序列 <400> 7 gtttccggta gcagatcccg ttcgctatgc cgttctgtac gaaaaccaga ccaggaatcc ccgctaccgc 60 agatggctta 120 aacagatcgc 180 tggttggacc ttcggtgctg gtgctgctct ccgtttcaac taaccagttc ggtatcggtg ttacccagaa aacaaagcta tctcccagat 29 1293957 ctgaccacca cctccaccgc tctgggtaaa ctgcaggacg ttgttaacca gaacgctcag 240 gctctgaaca ccctggttaa acagctgtcc taatag 276 <210> 8 <211> 309 <212> DNA <213> 人工合成 <220> <223〉 編碼有可於大腸桿菌表現系統中表現出嚴重急性 呼吸道症候群病毒E2棘蛋白之核酸序列 <400> 8 tccaacttcg ggacaaagtt gaagctgaag gcagacctac gttacccagc ggctgctacc aaaatgtccg taaaggttac gtgctatctc 60 ttcagatcga 120 agctgatccg 180 aatgcgttct 240 ctccgttctg ccgtctgatc tgctgctgaa gggtcagtcc aacgacatcc accggtcgtc atccgtgctt aaacgtgttg tgtcccgtct tgcagtccct ccgctaacct acttctgcgg 30 1293957 cacctgatgt ccttcccgca ggctgctccg cacggtgttg ttttcctgca cgttacctac 300 gtttaatag 309
31
Claims (1)
- 圆§7_u月赃頁 十、申請專利範圍： 1. 一種超抗原融合蛋白，係由以下胜肽片段所組成： 一段嚴重急性呼吸道症候群病毒之E2棘蛋白（E2 spike protein)胜肽片段； 一段假單胞菌屬外毒素（pseudomonas exotoxin)之胜肽片 段；以及 一段橋接該E2棘蛋白胜肽片段及該假單包菌屬外毒素胜 肽片段之胜肽； 其中，該嚴重急性呼吸道症候群病毒E2棘蛋白胜肽片段係 為 SEQ ID ΝΟ·3。 2. 如申請專利範圍第1項所述之超抗原融合蛋白，其中 該超抗原融合蛋白係藉由一大腸桿菌表現系統所產生。 3·如申請專利範圍第1項所述之超抗原融合蛋白，其中 該假早胞議屬外毋素包含一結合功此部位（binding domain) 與移位功能部位（translocation domain) 〇 4.如申請專利範圍第1項所述之超抗原融合蛋白，其中 該嚴重急性呼吸道症候群病毒之E2棘蛋白胜肽片段係為接 合於T細胞之胜肽。 5 · —種超抗原融合蛋白之醫藥組合物，係包含： 一超抗原融合蛋白，該超抗原融合蛋白係由一段嚴重急 性呼吸道症候群病毒之E2棘蛋白胜肽片段、一段假單胞菌屬 外毒素之胜肽片段、以及一段橋接該E2棘蛋白胜肽片段及該 假單包菌屬外毒素胜肽片段之胜肽所組成； 其中，該嚴重急性呼吸道症候群病毒E2棘蛋白胜肽片段係 32 1293957 為 SEQ ID Ν0·3。 6. 如申請專利範圍第5項所述之超抗原融合蛋白，其中 該超抗原融合蛋白係藉由一大腸桿菌表現系統所產生。 7. 如申請專利範圍第5項所述之超抗原融合蛋白，其中 該假單胞菌屬外毒素包含一結合功能部位（binding domain) 與移位功能部位（translocation domain) 〇 8. 如申請專利範圍第5項所述之超抗原融合蛋白，其中 該嚴重急性呼吸道症候群病毒之E2棘蛋白胜肽片段係為接 合於T細胞之胜肽。 9. 如申請專利範圍第5項所述之醫藥組合物，其係用於 治療嚴重急性呼吸道症候群病毒之感染。 10. 如申請專利範圍第5項所述之醫藥組合物，其更包括 醫藥上可接受之載體。 11. 如申請專利範圍第5項所述之醫藥組合物，其係用以 作為製造嚴重急性呼吸道症候群病毒之疫苗。
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| TW093121720A TWI293957B (en) | 2004-07-21 | 2004-07-21 | A superantigen fusion protein and the use thereof |
| JP2004327802A JP2006025782A (ja) | 2004-07-21 | 2004-11-11 | スーパー抗原融合蛋白及びその応用方法 |
| US11/183,796 US7618635B2 (en) | 2004-07-21 | 2005-07-19 | Super-antigen fusion proteins and the use thereof |
| CA002512515A CA2512515A1 (en) | 2004-07-21 | 2005-07-20 | Super-antigen fusion proteins and the use thereof |
| RU2005123050/13A RU2005123050A (ru) | 2004-07-21 | 2005-07-20 | Супер-антигенные фьюжн-белки и их использование |
| KR1020050066103A KR100801810B1 (ko) | 2004-07-21 | 2005-07-21 | 슈퍼-항원 융합 단백질들 및 그 용도 |
| AU2005203173A AU2005203173B2 (en) | 2004-07-21 | 2005-07-21 | Super-antigen fusion proteins and the use thereof |
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| GB0803076D0 (en) | 2008-02-20 | 2008-03-26 | Univ Ghent | Mucosal Membrane Receptor and uses thereof |
| US11246915B2 (en) | 2010-09-15 | 2022-02-15 | Applied Molecular Transport Inc. | Cholix toxin-derived fusion molecules for oral delivery of biologically active cargo |
| US11129906B1 (en) | 2016-12-07 | 2021-09-28 | David Gordon Bermudes | Chimeric protein toxins for expression by therapeutic bacteria |
| EP4082558B1 (en) | 2018-03-08 | 2023-08-23 | Applied Molecular Transport Inc. | Toxin-derived delivery constructs for oral delivery |
| CN113347997A (zh) | 2018-11-07 | 2021-09-03 | 应用分子运输公司 | 用于经口递送异源有效载荷的Cholix衍生的携带体 |
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| DE69629580D1 (de) * | 1995-10-13 | 2003-09-25 | Us Gov Health & Human Serv | Immunotoxin enthaltend ein disulfid-stabilisiertes antikörperfragment |
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| US7618635B2 (en) | 2009-11-17 |
| KR20060053966A (ko) | 2006-05-22 |
| RU2005123050A (ru) | 2007-01-27 |
| JP2006025782A (ja) | 2006-02-02 |
| KR100801810B1 (ko) | 2008-02-11 |
| CA2512515A1 (en) | 2006-01-21 |
| AU2005203173B2 (en) | 2007-08-09 |
| US20060134753A1 (en) | 2006-06-22 |
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