TWI781415B - 用於皮下注射之包含人類玻尿酸酶ph20變異體及藥物的醫藥組合物 - Google Patents

Abstract

本發明係關於一種醫藥組合物，其包含(a)藥物及(b)人類PH20變異體。根據本發明之醫藥組合物所包含之人類PH20變異體在選自在具有SEQ ID NO: 1之胺基酸序列的野生型人類PH20中之α螺旋8域(S347至C381)及介於α螺旋7與α螺旋8之間的連接域(A333至R346)中之一或多個區域包含胺基酸殘基取代，其中位於胺基端或羧基端的胺基酸殘基被選擇性地切除。此外，根據本發明之醫藥組合物可更包含藥學上可接受的添加劑，特別是穩定劑。根據本發明之醫藥組合物可因PH20變異體的效果而使與其結合之藥物的治療效果最大化。

Description

用於皮下注射之包含人類玻尿酸酶PH20變異體及藥物的醫藥組合物

本發明係關於醫藥組合物，包含具有提升的酵素活性及熱穩定性之人類玻尿酸酶(human hyaluronidase) PH20變異體及一或多種藥物，以及使用其治療疾病的方法。

根據本發明之醫藥組合物可較佳地使用於皮下注射。

應以高劑量或多劑量給藥的藥物，尤其是抗體藥物等藥物，一般透過靜脈注射給藥，此種注射需要約90分鐘或更久，靜脈注射伴隨額外的準備程序，因此對患者、醫生及醫護人員相當不便，且會產生額外的費用。相較之下，皮下注射具有能夠立即給藥的優點，但相較於靜脈注射，吸收速率相對較低，且當注射量為3-5毫升或更多時，因吸收緩慢，故在注射處可能會造成腫脹及疼痛。因此，蛋白質治療劑的皮下注射通常限於2毫升或更少的少量的溶液注射。然而，當玻尿酸酶與治療藥物一起皮下給藥(或皮下注射)時，分布於胞外基質的玻尿酸藉由玻尿酸酶的作用而被水解，因此皮下區域的黏度降低且物質的滲透性增加，因此，高劑量或多劑量的藥品可輕易輸送到體內。

人體中有六種玻尿酸酶基因：Hyal1、Hyal2、Hyal3、Hyal4、HyalPS1及PH20/SPAM1。Hyal1及Hyal2表現於大多數組織，PH20/SPAM1(以下稱為PH20)表現於精細胞膜及頂體膜(acrosomal membrane)。HyalPS1為偽基因所以不表現。PH20為切斷N-乙醯葡萄胺糖與葡萄醣醛酸之間的β-1,4鍵的酶(EC 3.2.1.35)，N-乙醯葡萄胺糖與葡萄醣醛酸為構成玻尿酸的糖。人類玻尿酸酶PH20的最佳pH值為5.5，但即使於pH 7-8亦展現出一些活性，而包含Hyal1之其他人類玻尿酸酶的最佳pH值為3-4，且於pH 7-8具有相當弱的活性。人體中皮下區域的pH值約為7.4，實質上為中性，因此，在多種類型的玻尿酸酶之中，PH20被廣泛應用於臨床使用。PH20之臨床使用的示例包含抗體治療劑的皮下注射、在眼科手術中作為眼部鬆弛劑(eye relaxant)及麻醉添加劑使用、藉由水解位於腫瘤細胞的胞外間質的玻尿酸用來增加抗癌治療劑進入腫瘤細胞以及用於促進組織中過量存在的體液及血液的吸收(resorption)。

同時，目前市售可得的PH20為從牛或羊的睪丸萃取的形式。其示例包含Amphadase® (牛玻尿酸酶)及Vitrase® (羊玻尿酸酶)。

牛睪丸玻尿酸酶(BTH)係透過在轉譯後修飾的過程中從牛野生型PH20的羧基端移除訊息肽及56個胺基酸而獲得。BTH亦為糖蛋白，並且基於其包含胺基酸之總組成，甘露糖含量為5%且葡萄糖胺含量為2.2% (Borders and Raftery, 1968)。當源自動物的玻尿酸酶以高劑量重複給予人體時，會產生中和抗體(neutralizing antibody)，而除了PH20之外還含有其他源自動物的生物材料作為不純物可能造成過敏反應。尤其，從牛萃取之PH20的用途因考量狂牛症而被限制。為了克服這些問題，已進行重組人類PH20蛋白的研究。

已報導將重組人類PH20蛋白表現於酵母菌(P. pastoris )、DS-2昆蟲細胞、動物細胞等 (Chen et al., 2016, Hofinger et al., 2007)。昆蟲細胞及酵母菌產生的重組PH20蛋白在轉譯後修飾的過程中的N-醣苷化模式方面不同於人類PH20。

在玻尿酸酶之中，已確定Hyal1 (PDB ID: 2PE4) (Chao et al., 2007)及蜂毒玻尿酸酶(bee venom hyaluronidase) (PDB ID: 1FCQ, 1FCU, 1FCV)的蛋白質結構。Hyal1由兩個結構域組成，催化結構域及類EGF結構域，催化結構域為α螺旋(alpha helix)及β股(beta-strand)各重複八次之(β/α)₈ 的形式(Chao et al., 2007)，其中α螺旋及β股表示蛋白質的二級結構。類EGF結構域在Hyal1之羧基端被不同地裁切之各變異體中為完全保留的(conserved)。Hyal1及PH20的胺基酸序列有35.1%相同，PH20的蛋白質三級結構尚未被發現。

在人類PH20的結構/功能關係的研究中，發現PH20的羧基端域對於蛋白質表現及酵素活性相當重要，尤其，已報導具有胺基酸477-483之羧基端的末端對於酵素表現及活性相當重要(Frost, 2007)。全長的PH20(胺基酸1-509)或在位置467羧基端被截斷之PH20變異體的活性僅為在位置477至483中之一個位點羧基端被截斷之PH20變異體的活性的10%或更低(Frost, 2007)。Halozyme Therapeutics發展出rHuPH20 (amino acids 36-482)，其為成熟的PH20在Y482羧基端被切斷的重組蛋白 (Bookbinder et al., 2006; Frost, 2007)。

同時，儘管正在進行研究以發展出使用人類PH20之皮下注射的形式的多種治療藥物，但人類PH20本身的低穩定性仍未解決。

在此技術背景下，本發明的發明人證實包含一或多個胺基酸殘基取代的人類PH20變異體具有非常高的酵素活性及熱穩定性，並因此提出專利申請案(PCT/KR 2019/009215)，其中一或多個胺基酸殘基取代位於野生型玻尿酸酶PH20之胺基酸序列中α螺旋8域(S347至C381)及介於α螺旋7及α螺旋8之間的連接域(A333至R346)，並且，在人類PH20變異體中位於PH20的胺基端及/或羧基端的一些胺基酸被切斷。

本發明之發明人亦證實根據本發明之PH20變異體可應用於醫藥組合物或包含藥物的製劑，藥物例如抗體藥物，尤其是高劑量的anti-HER2抗體或免疫檢查點抗體(immune checkpoint antibody)，故根據本發明之包含PH20變異體以及如anti-HER2抗體或免疫檢查點抗體之藥物的醫藥組合物及製劑可使用於皮下注射，如抗體藥物之藥物及PH20變異體的活性非常穩定且可長時間維持，進而完成本發明。

本發明鑑於前述問題而完成，本發明之一目的在於提供新穎的醫藥組合物，尤其是可使用於皮下注射的醫藥組合物，醫藥組合物包含具有提升的酵素活性及熱穩定性之PH20變異體及藥物，其中藥物及PH20變異體的熱穩定性及活性可長時間維持。

本發明另一目的在於提供治療疾病的方法，包含將根據本發明之醫藥組合物給予需要治療的受試者。

根據本發明，上述及其他目的可透過包含(a)藥物及(b)PH20變異體的醫藥組合物來完成。

根據本發明之醫藥組合物所包含之PH20變異體在具有SEQ ID NO: 1之胺基酸序列的野生型人類PH20中包含選自由S343E、M345T、K349E、L353A、L354I、N356E及I361T組成之群組之一或多個胺基酸殘基取代，並在選自α螺旋8域(S347至C381)及/或介於α螺旋7與α螺旋8之間的連接域(A333至R346)之一或多個區域中可更包含胺基酸殘基取代，其中位於胺基端及/或羧基端的一些胺基酸殘基被選擇性地切斷。

根據本發明之醫藥組合物可更包含選自藥學上可接受的添加劑之一或多者，尤其是緩衝液、穩定劑及界面活性劑。

根據本發明之醫藥組合物可以用於皮下注射之注射製劑的形式使用。

除非另有定義，否則本文中所使用之所有技術及科學術語具有如本發明所屬領域具有通常知識者所理解般相同的意義。一般而言，本文所使用之命名法為在本領域中習知且常用的。

本發明一實施例係關於醫藥組合物，包含(a)藥物及(b)PH20變異體，根據本發明之醫藥組合物可使用於預防或治療疾病，較佳地適用於皮下注射。

根據本發明之醫藥組合物所包含之PH20變異體在(具有SEQ ID NO: 1之胺基酸序列之)野生型PH20的胺基酸序列中，較佳地在(具有由SEQ ID NO: 1之胺基酸序列中之L36至S490組成的序列之)成熟的野生型PH20的胺基酸序列中，在對應α螺旋域及/或其連接域的區域中具有一些胺基酸殘基取代，較佳地在α螺旋8域(S347至C381)及/或介於α螺旋7與α螺旋8之間的連接域(A333至R346)，更佳地在T341至N363之中的胺基酸區域，最佳地在T341至I361、L342至I361、S343至I361、I344至I361、M345至I361或M345至N363。

在本發明中，「成熟的野生型PH20」係指包含之SEQ ID NO: 1之胺基酸殘基L36至S490的蛋白質，其缺少在具有SEQ ID NO: 1之序列的野生型PH20的胺基酸序列中形成訊息肽之M1至T35以及與PH20的實質功能無關之A491至L509。

表1，野生型PH20的胺基酸序列 (SEQ ID NO: 1) MGVLKFKHIFFRSFVKSSGVSQIVFTFLLIPCCLTLNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSIMRSMKSCLLLDNYMETILNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLSATMFIVSILFLIISSVASL

具體而言，根據本發明之醫藥組合物所包含之PH20變異體及其片段在具有SEQ ID NO: 1之序列的野生型PH20中包含一或多個突變，較佳地選自由S343E、M345T、K349E、L353A、L354I、N356E及I361T組成之群組之胺基酸殘基取代，最佳地選自由L354I及N356E組成之群組之一或多個胺基酸殘基取代。

在本發明中，「PH20變異體」之用語旨在包含一些胺基酸殘基的突變，較佳地在野生型人類PH20的序列中之胺基酸殘基的取代，以及與此胺基酸殘基的取代一起之在胺基端及/或羧基端的一些胺基酸殘基的缺失，並實質上與以「PH20變異體或其片段」之表示相同的意義使用。

本發明的發明人已證實，根據實驗結果，當在人類PH20的α螺旋8域及介於α螺旋7與α螺旋8之間的連接域的胺基酸序列部分被具有高親水性之Hyal1的α螺旋8域及介於α螺旋7及α螺旋8之間的連接域的胺基酸序列取代時，在中性pH下酵素活性及蛋白質聚集溫度(protein aggregation temperature，Tagg)會增加，透過此先前研究，可提供相較於野生型PH20而具有提高的酵素活性及熱穩定之新穎的PH20變異體及其片段性。

因此，根據本發明之醫藥組合物所包含之PH20變異體在(具有SEQ ID NO: 1之胺基酸序列之)野生型PH20的胺基酸序列中，較佳地在(具有由SEQ ID NO: 1之胺基酸序列中之L36至S490組成的序列之)成熟的野生型PH20的胺基酸序列中，包含選自由S343E、M345T、K349E、L353A、L354I、N356E及I361T組成之群組之一或多個胺基酸殘基取代，較佳地選自由L354I及N356E組成之群組之一或多個胺基酸殘基取代。

其中，在對應α螺旋域及/或其連接域的區域中，較佳地在α螺旋8域(S347至C381)及/或介於α螺旋7與α螺旋8之間的連接域(A333至R346)，更佳地在對應T341至N363、T341至I361、L342至I361、S343至I361、I344至I361、M345至I361或M345至N363之胺基酸區域中，有一或多個胺基酸殘基被取代。

具體而言，在根據本發明之醫藥組合物所包含之PH20變異體中，在野生型PH20中，較佳地在成熟的野生型PH20中，α螺旋8域(S347至C381)及/或介於α螺旋7及α螺旋8之間的連接域(A333至R346)可被一些胺基酸殘基取代，其為具有SEQ ID NO: 51之序列(請見表2及表3)的Hyal1之對應區域的胺基酸序列中之一些胺基酸殘基，但本發明不限於此。

表2，野生型Hyal1 (SEQ ID NO: 51)的胺基酸序列 MAAHLLPICALFLTLLDMAQGFRGPLLPNRPFTTVWNANTQWCLERHGVDVDVSVFDVVANPGQTFRGPDMTIFYSSQLGTYPYYTPTGEPVFGGLPQNASLIAHLARTFQDILAAIPAPDFSGLAVIDWEAWRPRWAFNWDTKDIYRQRSRALVQAQHPDWPAPQVEAVAQDQFQGAARAWMAGTLQLGRALRPRGLWGFYGFPDCYNYDFLSPNYTGQCPSGIRAQNDQLGWLWGQSRALYPSIYMPAVLEGTGKSQMYVQHRVAEAFRVAVAAGDPNLPVLPYVQIFYDTTNHFLPLDELEHSLGESAAQGAAGVVLWVSWENTRTKESCQAIKEYMDTTLGPFILNVTSGALLCSQALCSGHGRCVRRTSHPKALLLLNPASFSIQLTPGGGPLSLRGALSLEDQAQMAVEFKCRCYPGWQAPWCERKSMW

表3，PH20及Hyal1之胺基酸序列之間的比較

α螺旋	PH20的胺基酸序列	Hyal1的胺基酸序列
α螺旋1	P56至D65	N39至G48
α螺旋3	S119至M135	S101至I117
α螺旋4'	K161至N176	K144至H159
α螺旋4	S180至R211	P163至R194
α螺旋5	F239至S256	P222至S239
α螺旋6	A274至D293	K257至G277
α螺旋7	S317至G332	P299至G314
α螺旋8	S347至C381	T329至C363

更具體而言，根據本發明之醫藥組合物所包含之PH20變異體或其片段在野生型PH20的胺基酸序列中，較佳地在成熟的野生型PH20的胺基酸序列中，較佳地包含L354I及/或N356E之胺基酸殘基取代；並且，較佳地更包含在選自T341至N363之一或多個位置的胺基酸殘基取代，較佳地在選自由T341、L342、S343、I344、M345、S347、M348、K349、L352、L353、D355、E359、I361及N363組成之群組之一或多個位置，但本發明不限於此；並且，更佳地更包含選自由T341S、L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、D355K、E359D、I361T及N363G組成之群組之一或多個胺基酸殘基取代，但本發明不限於此。

較佳地，根據本發明之醫藥組合物所包含之PH20變異體可包含選自M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T之胺基酸殘基取代；並且，可更包含選自由T341S、L342W、S343E、I344N及N363G組成之群組之一或多個胺基酸殘基取代，但本發明不限於此。

更佳地，根據本發明之醫藥組合物所包含之PH20變異體或其片段可包含，但不限於，選自以下群組之任一取代： (a) T341S、L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T； (b) L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T； (c) M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T； (d) M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D、I361T及N363G； (e) I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T；以及 (f) S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D以及I361T。

在本發明中，以一個字母之胺基酸殘基代號與數字一起的表達方式，例如「S347」，表示胺基酸殘基在SEQ ID NO: 1之胺基酸序列中對應的位置。

舉例而言，「S347」表示在SEQ ID NO: 1之胺基酸序列中之位置347的胺基酸殘基為絲胺酸(serine)。此外，「S347T」表示在SEQ ID NO: 1之位置347的絲胺酸被取代成蘇胺酸(threonine)。

根據本發明之醫藥組合物所包含之PH20變異體被解釋為包含在特定胺基酸殘基位置的胺基酸殘基被保留地取代(conservatively substituted)的變異體。

如本文所使用之「保留的取代(conservative substitution)」之用語係指PH20變異體的修飾，其涉及具有相似生化性質的一或多個胺基酸的取代，其並不會導致失去對應之PH20變異體的生物或生化功能。

「保留的胺基酸取代」係胺基酸殘基被具有相似的側鏈的胺基酸殘基取代者。具有相似的側鏈的胺基酸殘基家族已被定義且為本領域習知的。這些家族包含具有鹼性側鏈的胺基酸(例如離胺酸、精胺酸及組胺酸)、具有酸性側鏈的胺基酸(例如天冬胺酸及麩胺酸)、具有不帶電極性側鏈的胺基酸(例如甘胺酸、天冬醯胺酸、麩醯胺酸、絲胺酸、蘇胺酸、酪胺酸及半胱胺酸)、具有非極性側鏈的胺基酸(例如丙胺酸、纈胺酸、白胺酸、異白胺酸、脯胺酸、苯丙胺酸、甲硫胺酸及色胺酸)、具有β支鏈側鏈的胺基酸(例如蘇胺酸、纈胺酸及異白胺酸)以及具有芳香族側鏈的胺基酸(例如酪胺酸、苯丙胺酸、色胺酸及組胺酸)。

可預期儘管具有保留的胺基酸取代，根據本發明之醫藥組合物所包含之PH20變異體仍保有其活性。

此外，根據本發明之醫藥組合物所包含之PH20變異體或其片段被解釋為包含PH20變異體或其片段，其具有與根據本發明之PH20變異體或其片段功能或作用實質上相同的功能及/或作用的，且具有與根據本發明之PH20變異體或其片段至少80%或85%之胺基酸序列同源性(amino acid sequence homology)，較佳地至少90%，更佳地至少95%，最佳地至少99%。

根據本發明之PH20變異體具有在動物細胞中增加的表現程度以及增加的蛋白質摺疊速率，從而相較於成熟的野生型PH20而具有增加的熱穩定性。此外，儘管熱穩定性增加，PH20變異體的酵素活性仍超過或相似於野生型PH20的酵素活性。

同時，已知當在成熟的野生型PH20的羧基端的一些胺基酸，例如S490，額外地被切除時，酵素活性會降低，但根據本發明之PH20變異體在即使成熟的野生型PH20的羧基端具有額外地被切除的序列時，相較於成熟的野生型PH20仍顯現出提升的熱穩定性及提升或相似的酵素活性。此外，最多五個胺基酸殘基從胺基端的胺基酸被切除時，PH20變異體仍保持其酵素活性，這表示從胺基端之P41起的殘基在蛋白質表現及酵素活性上有重要的作用。

因此，根據本發明之醫藥組合物所包含之PH20變異體在野生型PH20之α螺旋8域(S347至C381)及/或介於α螺旋7及α螺旋8之間的連接域(A333至R346)中包含一些胺基酸殘基取代，並在羧基端及/或胺基端更包含一些胺基酸殘基缺失，但本發明不限於此。

在一些實施例中，根據本發明之醫藥組合物所包含之PH20變異體可包含在胺基端的一些胺基酸殘基缺失及/或在羧基端的一些胺基酸缺失，其中在胺基端的一些胺基酸殘基缺失係源自在選自由SEQ ID NO: 1之胺基酸序列的胺基端之M1至P42組成的群組的胺基酸殘基之前的切除(cleavage)，較佳地在胺基酸殘基L36、N37、F38、R39、A40、P41或P42之前，而在羧基端的一些胺基酸缺失係源自在選自由羧基端之V455至W509組成之群組之胺基酸殘基之後的切除，較佳地在選自由V455至S490組成之群組之後，最佳地在V455、C458、D461、C464、I465、D466、A467、F468、K470、P471、P472、M473、E474、T475、E476、P478、I480、Y482、A484、P486、T488或S490之後。

「在胺基端之L36、N37、F38、R39、A40、P41或P42之前的切除」之說法分別表示在SEQ ID NO: 1之胺基酸序列中緊接L36之前從M1至T35的所有胺基酸殘基、緊接N37之前從M1至L36的所有胺基酸殘基、緊接F38之前從M1至N37的所有胺基酸殘基、緊接R39之前從M1至F38的所有胺基酸殘基、緊接A40之前從M1至R39的所有胺基酸殘基、緊接P41之前從M1至A40的所有胺基酸殘基或緊接P42之前從M1至P41的所有胺基酸殘基被切斷並移除。「在SEQ ID NO: 1之胺基端之M1之前的切除」之說法表示在胺基端沒有發生任何切除。

此外，「在羧基端之V455、C458、D461、C464、I465、D466、A467、F468、K470、P471、P472、M473、E474、T475、E476、P478、I480、Y482、A484、P486、T488或S490之後的切除」之說法分別表示切斷並移除在SEQ ID NO: 1之序列中V455、C458、D461、C464、I465、D466、A467、F468、K470、P472、M473、E474、T475、E476、P478、I480、Y482、A484、P486、T488或S490之後的胺基酸殘基。舉例而言，在S490的切除表示在S490及A491之間的切除。

較佳地，根據本發明之醫藥組合物所包含之人類PH20變異體可具有選自由SEQ ID NOS: 5至50之胺基酸序列組成之群組之胺基酸序列，較佳地具有SEQ ID NO: 44之胺基酸序列，但本發明不限於此。在根據本發明之特定實施例中所建構的PH20變異體中，經取代或切除的胺基酸的序列如以下表4所示。

表4，根據本發明之PH20變異體的胺基酸序列及其取代/切除特性

名稱	序列號	取代	序列
HM1	5	12個胺基酸被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D、I361T及N363G。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T L G PYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM2	6	7個胺基酸被取代為Y365F、I367L、L371S、A372G、K374L、M375L及V379A。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSITRTKESCQAIKEYMDTTLNP F I L NVT SG A LL CSQ A LCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM3	7	19個胺基酸被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D、I361T、N363G、Y365F、I367L、L371S、A372G、K374L、M375L及V379A。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T L G P F I L NVT SG A LL CSQ A LCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM4	8	17個胺基酸被取代為G340V、T341S、L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D、I361T及N363G。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIW VSWENT R TKE SC QAIKE YM D T T L G PYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM6	9	11個胺基酸殘基被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM7	10	16個胺基酸被取代為G340V、T341S、L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIW VSWENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM8	11	12個胺基酸被取代為I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLS NT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM9	12	13個胺基酸被取代為S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTL ENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HMl0	13	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HMl1	14	13個胺基酸殘基被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D、I361T、Y365F及I367L。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNP F I L NVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM12	15	15個胺基酸殘基被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D、I361T、Y365F、I367L、L371S及A372G。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNP F I L NVT SG AKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM13	16	11個胺基酸殘基被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及 I361T，並在胺基端之殘基F38之前進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM14	17	11個胺基酸被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並在I465之羧基之後進行切除。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCI
HM15	18	11個胺基酸被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並在F468之羧基之後進行切除。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAF
HM16	19	11個胺基酸被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並在P471之羧基之後進行切除。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKP
HM17	20	胺基酸L36至V47被取代為FRGPLLPNR，並且11個胺基酸被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。	FRGPLLPNR PFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM18	21	胺基酸L36至A52被取代為FRGPLLPNRPFTTV，並且11個胺基酸被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。	FRGPLLPNRPFTTV WNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM19	22	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基K470之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLK
HM20	23	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基F468之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAF
HM21	24	15個胺基酸殘基被取代為T341S、L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWG SWENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM24	25	11個胺基酸殘基被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基 A40之前進行切除。	APPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM25	26	11個胺基酸被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基P42之前進行切除。	PVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM29	27	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基L36之前以及在羧基端之殘基A467之後進行切除。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDA
HM30	28	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基L36之前以及在羧基端之殘基 C464之後進行切除。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVC
HM31	29	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基 L36之前以及在羧基端之殘基 D461之後進行切除。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIAD
HM32	30	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基L36之前以在羧基端之殘基C458之後進行切除。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVC
HM33	31	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基L36之前以及羧基端之殘基V455之後進行切除。	LNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAV
HP34	32	15個胺基酸殘基被取代為T341S、L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基K470之後 進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWG SWENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLK
HM35	33	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基P472之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPP
HM36	34	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基M473之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPM
HM37	35	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基E474之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPME
HM38	36	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基T475之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMET
HM39	37	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基E476之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETE
HM40	38	11個胺基酸殘基被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基N37之前進行切除。	NFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM41	39	11個胺基酸殘基被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基R39之前進行切除。	RAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM42	40	11個胺基酸殘基被取代為M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基P41之前進行切除。	PPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSI T R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS
HM43	41	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基I465之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGS WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCI
HM44	42	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及以及在羧基端之殘基D466之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGS WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCID
HM45	43	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及以及在羧基端之殘基A467之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGS WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDA
HP46	44	15個胺基酸殘基被取代為T341S、L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基F468之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWG SWENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAF
HM47	45	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基P478之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEP
HM48	46	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基I480之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQI
HM49	47	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基Y482之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFY
HM50	48	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及以及在羧基端之殘基A484之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNA
HM51	49	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基P486之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASP
HM52	50	14個胺基酸殘基被取代為L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T，並且在胺基端之殘基F38之前以及在羧基端之殘基T488之後進行切除。	FRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGT WENT R TKE SC QAIKE YM D T T LNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPST

同時，先前研究報導野生型PH20的酵素活性會依據位於羧基端之胺基酸殘基的切除位置而改變。然而，在本發明中，形成PH20之二級結構之特定的α螺旋被人類玻尿酸酶的α螺旋取代，從而建構出相較於野生型PH20而言具有較高之穩定性的PH20變異體，並且在這些變異體中，被取代的α螺旋結構域與PH20的其他二級結構之間的相互作用顯現出不同於野生型PH20之模式的模式，使得此變異體不論羧基端的切除位置，皆具有某種程度或較高的酵素活性。

此外，在本發明中，試圖藉由使用在動物細胞中展現高蛋白表現程度的其他蛋白質的訊息肽取代原本的人類PH20的訊息肽，來增加重組PH20蛋白的表現。

因此，在另一實施例中，根據本發明之醫藥組合物所包含之PH20變異體在其胺基端可包含源自人類玻尿酸酶-1(human hyaluronidase-1，Hyal1)、人類生長激素或人類血清白蛋白的訊息肽，取代由M1至T35組成之野生型PH20的訊息肽，並較佳地可包含具有根據SEQ ID NO: 2之MATGSRTSLLLAFGLLCLPWLQEGSA之胺基酸序列的源自人類生長激素的訊息肽、具有根據SEQ ID NO: 3之MKWVTFISLLFLFSSAYS之胺基酸序列的源自人類血清白蛋白的訊息肽或具有根據SEQ ID NO: 4之MAAHLLPICALFLTLLDMAQG之胺基酸序列的源自人類Hyal1的訊息肽，如表5所示，但本發明不限於此。

表5，人類生長激素、人類血清白蛋白或人類Hyal1的訊息肽的胺基酸序列

訊息肽的來源	胺基酸序列	SEQ ID NO:
人類生長激素	MATGSRTSLLLAFGLLCLPWLQEGSA	2
人類血清白蛋白	MKWVTFISLLFLFSSAYS	3
人類Hyal1	MAAHLLPICALFLTLLDMAQG	4

在根據本發明之醫藥組合物所包含之PH20變異體之中，具有組胺酸標籤(6xHis-tag)附接於羧基端的變異體命名為HM，沒有組胺酸標籤的變異體命名為HP。此外，具有組胺酸標籤附接於羧基端之成熟的野生型PH20 (L36-S490)命名為WT，沒有組胺酸標籤且羧基端在Y482之後被切除的成熟的野生型PH20 (L36至Y482)命名為HW2。

HP46 (SEQ ID NO: 44)係藉由以下方式而獲得的PH20變異體：使用具有已知的蛋白質三級結構的人類玻尿酸酶之Hyal1 (PDB ID: 2PE4)來模擬(modeling)蛋白質結構(Chao et al., 2007)，接著以Hyal1的胺基酸序列取代α螺旋8及介於α螺旋7與α螺旋8之間的連接域之胺基酸的胺基酸序列，並使胺基端在F38被切除且使羧基端在F468之後被切除。尤其，α螺旋8位於PH20的蛋白質三級結構之外，且相較於其他α螺旋而言與相鄰的α螺旋或β股具有較少的交互作用。一般而言，酵素活性及熱穩定性在其之間具有權衡關係(trade-off relationship)，故蛋白質的熱穩定性愈高，酵素活性愈低，而當酵素活性因蛋白質結構的靈活性(flexibility)改善而增加時，熱穩定性會傾向降低。然而，在pH7下由濁度法(Turbidimetric assay)量測之HP46的特定活性約為46單位/微克，其被評估約為野生型PH20的特定活性(約為23單位/微克)的兩倍。

蛋白質的熱穩定性可依據50%之蛋白質三級結構變性(denature)的熔點(melting temperature，Tm)以及蛋白質之間發生聚集的聚集溫度(aggregation temperature，Tagg)來評估。一般而言，蛋白質的聚集溫度傾向低於其熔點。相較於PH20的α螺旋8而言，Hyal1的α螺旋8展現出較高的親水性。經取代之Hyal1的α螺旋8提高HP46的蛋白質表面親水性，從而造成因疏水性作用所致之蛋白質之間的聚集效應延緩，故聚集溫度為51°C，其相較於野生型PH20的聚集溫度(46.5°C)而言，觀察到4.5°C的增加。

HP46係在α螺旋8及介於α螺旋7與α螺旋8之間的連接域中的胺基酸殘基被取代的變異體，其中T341被絲胺酸取代。當胺基酸殘基341為蘇胺酸，酵素活性相似於野生型PH20的酵素活性，但當取代為絲胺酸時，酵素活性增加約2倍，並證實即使在受質凝膠分析(substrate gel assay)中，相較於野生型PH20而言，所得之變異體水解5至6倍的玻尿酸。受質凝膠分析涉及蛋白質變性(denaturation)及再摺疊(refolding)過程，其代表相較於野生型PH20而言，HP46的蛋白質三級結構的再摺疊及恢復受到提升。

在根據本發明之醫藥組合物中PH20變異體的量微至少為50單位/毫升，較佳地在100單位/毫升至20,000單位/毫升的範圍內，更佳地在約150單位/毫升至約18,000單位/毫升的範圍內，又更佳地在1,000單位/毫升至16,000單位/毫升的範圍內，最佳地在1,500單位/毫升至12,000單位/毫升的範圍內。

根據本發明之醫藥組合物所包含之藥物的示例包含，但不限於，蛋白質藥物、抗體藥物、小分子、適體、RNAi、反義RNA及細胞治療劑，例如嵌合抗原受體T細胞(chimeric antigen receptor T-cell，CAR-T)或嵌合抗原受體自然殺手細胞(CAR-natural killer，CAR-NK)，其不僅可使用目前市售可得的藥物，亦可使用在臨床試驗中或開發中的藥物。

作為藥物，較佳地可使用蛋白質藥物或抗體藥物。

根據本發明之醫藥組合物所包含之「蛋白質藥物」係由胺基酸組成的藥物，故透過蛋白質的活性而展現出治療或防止疾病的功效，且係由不同於抗體藥物的蛋白質組成之藥物，並可選自由細胞介素(cytokine)、治療性酵素、激素(hormone)、可溶性受體及其融合蛋白、胰島素或其類似物、骨成形性蛋白質(bone morphogenetic protein，BMP)、紅血球生成素(erythropoietin)及血清衍生蛋白(serum-derived protein)組成之群組，但本發明不限於此。

根據本發明之醫藥組合物所包含之細胞介素可選自由干擾素(interferon)、介白素(interleukin)、群落刺激因子(colony-stimulating factor，CSF)、腫瘤壞死因子(tumor necrosis factor，TNF)及組織生長因子(tissue growth factor，TGF)組成之群組，但本發明不限於此。

治療性酵素可包含，但不限於，β葡萄糖腦苷酶(β-glucocerebrosidase)及阿加糖酶(agalsidase β)。

根據本發明之醫藥組合物所包含之可溶性受體係受體的細胞外結構域(extracellular domain)，其融合蛋白係抗體的Fc區域或類似者融合至可溶性受體的蛋白質。可溶性受體為與疾病相關之配體所結合之受體的可溶形式，其示例包含Fc區域融合至TNF-α可溶性受體的形式(例如包含etanercept成分及相似於其之形式的產品)、Fc區域融合至血管內皮細胞生長因子(VEGF)可溶性受體的形式(例如包含alefacept成分及相似於其之形式的產品)、Fc區域融合至CTLA-4的形式(例如包含abatacept或belatacept成分及相似於其形式之產品)、Fc區域融合至介白素1可溶性受體的形式(例如包含rilonacept成分及相似於其形式之產品)以及Fc區域融合至LFA3可溶性受體的形式(例如包含alefacept成分及相似於其形式之產品)，但本發明不限於此。

根據本發明之醫藥組合物所包含之激素係指用於治療或預防由激素缺乏等所致之疾病而注射至人體內的激素或其類似物，其示例包含，但不限於，人類生長激素、雌激素(estrogen)及黃體素(progesterone)。

根據本發明之醫藥組合物所包含之血清衍生蛋白係存在於血漿的蛋白質，並包含從血漿中萃取的蛋白質及產生的重組蛋白，其示例可包含，但不限於血纖維蛋白原(fibrinogen)、溫韋伯氏因子(von Willebrand factor)、白蛋白、凝血酶、因子II (FII)、因子V (FV)、因子VII (FVII)、因子IX (FIX)、因子X (FX)及因子 (FXI)。

根據本發明之醫藥組合物所包含之抗體藥物可為單株抗體藥物或多株抗體藥物。

根據本發明之單株抗體藥物係包含單株抗體及單株抗體片段的蛋白質，其中單株抗體及單株抗體片段能夠特異地結合至與特定疾病相關之抗原。單株抗體亦包含雙特異性抗體，包含單株抗體或其片段的蛋白質概念上包含抗體藥物複合體(antibody-drug conjugate，ADC)。

與特定疾病相關之抗原的示例包含4-1BB、整合素(integrin)、類澱粉蛋白β(amyloid beta)、血管生長素(angiopoietin) (血管生長素1或2)、血管生長素類似物3、B細胞活化因子(B-cell-activating factor，BAFF)、B7-H3、補體5(complement 5)、CCR4、CD3、CD4、CD6、CD11a、CD19、CD20、CD22、CD30、CD33、CD38、CD52、CD62、CD79b、CD80、CGRP、密連蛋白-18(Claudin-18)、補體因子D(complement factor D)、CTLA4、DLL3、EGF受體、血友病因子(hemophilia factor)、Fc受體、FGF23、葉酸受體(folate receptor)、GD2、GM-CSF、HER2、HER3、干擾素受體、干擾素γ、IgE、IGF-1受體、介白素1、介白素2受體、介白素4受體、介白素5、介白素5受體、介白素6、介白素6受體、介白素7、介白素12/23、介白素13、介白素17A、介白素17受體 A、介白素31受體、介白素36受體、LAG3、LFA3、NGF、PVSK9、PD-1、PD-L1、RANK-L、SLAMF7、組織因子、TNF、VEGF、VEGF受體及溫韋伯氏因子(von Willebrand factor，vWF)，但本發明不限於此。

以下為，但不限於，包含針對與特定疾病相關之單株抗體或單株抗體片段的蛋白質： utomilumab作為抗4-1BB抗體； natalizumab、etrolizumab、vedolizumab及bimagrumab作為針對整合素的抗體； bapineuzumab、crenezumab、solanezumab、aducanumab及gantenerumab作為針對類澱粉蛋白β的抗體； 針對血管生長素的抗體，例如針對血管生長素1及2之AMG780、MEDI 3617及針對血管生長素2之nesvacumab，以及針對血管生長素2及VEGF為雙特異性抗體之vanucizumab； Evinacumab作為針對血管生長素類似物3的抗體； tabalumab、lanalumab、and belimumab作為針對細胞活化因子(BAFF)的抗體； omburtamab作為B7-H3的抗體； ravulizumab及eculizumab作為針對補體5的抗體； mogamulizumab作為針對CCR4的抗體； otelixizumab、teplizumab及muromonab作為針對CD3的抗體，tebentafusp作為針對GP100及CD3的雙特異性抗體，blinatumomab作為針對CD19及CD3的雙特異性抗體，及REGN1979作為針對CD20及CD3的雙特異性抗體； ibalizumab及zanolimumab作為針對CD4的抗體； itolizumab作為針對CD6的抗體； efalizumab作為針對CD11a的抗體； inebilizumab、tafasitamab及loncastuximab tesirine(其為抗體藥物複合體)作為針對CD19的抗體； ocrelizumab、ublituximab、obinutuzumab、ofatumumab、rituximab、tositumomab、及ibritumomab tiuxetan(其為抗體藥物複合體)作為針對CD20的抗體； epratuzumab、inotuzumab ozogamicin(其為抗體藥物複合體)及moxetumomab pasudotox作為針對CD22的抗體； brentuximab vedotin作為針對的CD30的抗體藥物複合體； vadastuximab talirine及gemtuzumab ozogamicin作為針對CD33的抗體藥物複合體； daratumumab及isatuximab作為針對CD38的抗體藥物複合體； alemtuzumab作為針對CD52的抗體； crizanlizumab作為針對CD62的抗體； polaruzumab vedotin作為針對CD79b的抗體藥物複合體； galiximab作為針對CD80的抗體； eptinezumab、fremanezumab、galcanezumab及erenumab作為針對CGRP的抗體； zolbetuximab作為針對密連蛋白-18的抗體； lampalizumab作為針對補體因子D的抗體； tremelimumab、zalifrelimab及ipilimumab作為針對CTLA4的抗體； rovalpituzumab tesirine作為針對DLL3的抗體藥物複合體； cetuximab、depatuxizumab、zalutumumab、necitumumab及panitumumab作為針對EGF受體的抗體； emicizumab作為針對凝血因子IX及因子X(其為血友病因子)的雙特異性抗體； nipocalimab及rozanolixizumab 作為針對Fc受體的抗體； burosumab作為針對FGF23的抗體； farletuzumab作為針對葉酸受體的抗體，以及mirvetuximab soravtansine作為針對葉酸受體的抗體藥物複合體； dinutuximab及naxitamab作為針對GD2的抗體； otilimab作為針對GM-CSF的抗體； margetuximab、pertuzumab及trastuzumab作為針對HER2的抗體，以及trastuzumab deruxtecan、trastuzumab emtansine及trastuzumab duocarmazine作為針對HER2的抗體藥物複合體； patritumab作為針對HER3的抗體； anifrolumab作為針對干擾素受體的抗體； emapalumab作為針對干擾素γ的抗體； ligelizumab及omalizumab作為針對IgE的抗體； dalotuzumab、figitumumab及teprotumumab作為針對IGF-1受體的抗體； gebokizumab及canakinumab作為針對介白素1的抗體； daclizumab及basiliximab作為針對介白素2受體的抗體； dupilumab作為針對介白素4受體的抗體； mepolizumab及reslizumab作為針對介白素5的抗體； benralizumab作為針對介白素5受體的抗體； clazakizumab、olokizumab、sirukumab及siltuximab作為針對介白素6的抗體； sarilumab、satralizumab、tocilizumab及REGN88作為針對介白素6受體的抗體； secukinumab作為針對介白素7的抗體； ustekinumab及briakinumab作為針對介白素12/23的抗體； lebrikizumab及tralokinumab作為針對介白素13的抗體； ixekizumab及bimekizumab作為針對介白素17A的抗體； brodalumab作為針對介白素17受體A的抗體； brazikumab、guselkumab、risankizumab、tildrakizumab及mirikizumab作為針對介白素23的抗體； nemolizumab作為針對介白素31受體的抗體； spesolimab作為針對介白素36受體的抗體； relatlimab作為針對LAG3的抗體； narsoplimab作為針對NASP2的抗體； fasinumab及tanezumab作為針對NGF的抗體； alirocumab、evolocumab及bococizumab作為針對PVSK9的抗體； lambrolizumab、balstilimab、camrelizumab、cemiplimab、dostarlimab、prolgolimab、shintilimab、spartalizumab、tislelizumab、pembrolizumab及nivolumab作為針對PD-1的抗體； atezolizumab、avelumab、envafolimab及durvalumab作為針對PD-L1的抗體，以及bintrafusp alpha作為針對TGF beta及PD-L1的雙特異性抗體； denosumab作為針對RANK-L的抗體； elotuzumab作為針對SLAMF7的抗體； concizumab及marstacimab作為針對組織因子的抗體； 針對TNF的抗體，尤其是TNFα，包含infliximab、adalimumab、golimumab、抗體片段certolizumab pegol及ozoralizumab(其為針對TNF及白蛋白的雙特異性抗體)； 針對VEGF的抗體，包含brolucizumab、ranibizumab、bevacizumab及faricimab(其為針對VEGF及Ang2的雙特異性抗體)； ramucirumab作為針對VEGF受體的抗體；以及 caplacizumab作為針對vWF的抗體

同時，在約20-25%之乳癌患者中觀察到人類表皮生長因子受體(human epidermal growth factor receptor 2，HER2)的過度表現，其促進細胞分裂，HER2過度表現的乳癌進展快速，具侵略性，且相較於HER2低表現的乳癌而對於化療的響應低，故其預後(prognosis)為不利的。Trastuzumab為靶向(targeting) HER2的單株抗體藥物，其特異性地結合於過度表現HER2的癌細胞的表面上的HER2，以抑制細胞複製及增生的訊息傳遞，從而減緩腫瘤的進展。Trastuzumab已在1998年經美國食品藥物管理局(United States Food and Drug Administration，FDA)核准在美國用於治療乳癌，並在2003年經韓國食品藥物管理局(KFDA)核准。從那時起，trastuzumab的功效在HER2過度表現的胃癌中亦得到認可，因此已作為胃癌的治療劑使用。

Roche的賀癌平皮下注射製劑(商品名：Herceptin)由440毫克之trastuzumab作為主要成分而組成，冷凍乾燥的trastuzumab與生理食鹽水混合並注射至靜脈。另一方面，trastuzumab的皮下注射製劑(商品名：賀癌平 SC)係5毫升之液體製劑，並包含600毫克(120毫克/毫升)之trastuzumab作為主要成分，並包含20 mM組胺酸(pH 5.5)、210 mM海藻糖、10 mM甲硫胺酸、0.04%聚山梨醇酯20及10,000單位之rHuPH20 (2,000 單位/毫升，0.004%，40微克/毫升)作為添加物。

賀癌平皮下注射製劑的保存期為21個月。Trastuzumab之靜脈注射製劑為冷凍乾燥形式且具有30個月的保存期，但trastuzumab之皮下注射製劑為液體狀態且具有較短之21個月的保存期。為此，可估計一或多個trastuzumab及重組人類玻尿酸酶PH20在液體製劑中的穩定性受到限制。

在本文中，在本發明中，就根據本發明之PH20變異體的特性的觀點而言，相較於野生型人類玻尿酸酶PH20及可從Halozyme獲得之重組人類PH20而言，PH20變異體不僅具有增加的酵素活性，亦具有高的蛋白質聚集溫度，故展現出提升的熱穩定性，皮下注射製劑的保存期定為長期，較佳地21個月或更久。

根據本發明之醫藥組合物中，抗體藥物的含量可為5毫克/毫升至 500毫克/毫升之範圍內，較佳地20毫克/毫升至200毫克/毫升，更佳地100毫克/毫升至150毫克/毫升，最佳地120 ± 18 毫克/毫升，舉例而言，約110毫克/毫升，約120毫克/毫升，或約130毫克/毫升。

根據本發明之醫藥組合物所包含之多株抗體較佳地為從血清萃取的血清抗體，例如免疫球蛋白，但不限於此。

在小分子化合物的情況下，可不受限制地使用對於治療或預防需要快速效果的任何藥物。舉例而言，可使用基於嗎啡的止痛劑(morphine-based painkiller) (Thomas et al., 2009)。此外，當作為用於由抗癌藥物所致之組織壞死的治療劑時，小分子化合物可單獨使用或與解毒藥物(例如長春花屬生物鹼(Vinca alkaloids)及紫杉烷(Taxanes))結合使用(Kreidieh et al., 2016)。

根據本發明之醫藥組合物可更包含由緩衝液、穩定劑及界面活性劑組成之群組之一或多者。

本發明之醫藥組合物所包含之緩衝液可不受限制地使用，只要能夠實現pH 4至8，較佳地為5至7，緩衝液較佳地選自由蘋果酸鹽、甲酸鹽、檸檬酸鹽、乙酸鹽、丙酸鹽、吡啶、哌𠯤、二甲砷酸鹽、琥珀酸鹽、2-(N-𠰌啉基)乙磺酸(MES)、組胺酸、三羥甲基胺基甲烷(Tris)、雙-(2-羥基乙基)胺基三羥甲基胺基甲烷(bis-Tris)、磷酸鹽、乙醇胺、碳酸鹽、哌𠯤-N,N'-雙(2-乙磺酸)(PIPES)、咪唑、1,3-雙(三(羥甲基)甲基胺基)丙烷(BIS-TRIS propane)、N,N-雙(2-羥乙基)-2-胺基乙磺酸(BES)、3-(N-𠰌啉基)丙磺酸(MOPS)、羥乙基哌𠯤乙磺酸(HEPES)、焦磷酸鹽及三乙醇胺組成之群組之一或多者，較佳地為組胺酸緩衝液，例如L-組胺酸/HCl，但不限於此。

緩衝液的濃度可為0.001 mM至200 mM之範圍，較佳地1 mM至50 mM，更佳地5 mM至40 mM，最佳地10 mM至30 mM。

在根據本發明之醫藥組合物中，穩定劑可不受限制地使用，只要他們通常在本領域中用於穩定蛋白質，較佳地，穩定劑例如可為選自由碳水化合物、糖或其水合物、糖醇或其水合物及胺基酸組成之群組之一或多者。

作為穩定劑使用之碳水化合物、糖或糖醇可為選自由海藻糖或其水合物、蔗糖、糖精、甘油、丁四醇、蘇糖醇、木糖醇、阿拉伯糖醇、核糖醇、甘露醇、山梨醇、半乳糖醇、海藻糖醇、艾杜糖醇、肌醇、庚七醇、異麥芽酮糖醇、麥芽糖醇、聚醣醇、環糊精、羥丙基環糊精及葡萄糖組成之群組之一或多者，但不限於此。

胺基酸可為選自由麩醯胺酸、麩胺酸、甘胺酸、離胺酸、離胺基離胺酸(lysilysine)、白胺酸、甲硫胺酸、纈胺酸、絲胺酸、硒甲硫胺酸、瓜胺酸、精胺酸、天冬醯胺酸、天冬胺酸、烏胺酸、異白胺酸、牛磺酸、茶胺酸、蘇胺酸、色胺酸、酪胺酸、苯丙胺酸、脯胺酸、吡咯離胺酸、組胺酸及丙胺酸組成之群組之一或多者，但不限於此。

在根據本發明之醫藥組合物中，作為穩定劑使用之糖或糖醇的濃度可為0.001 mM至500 mM之範圍，較佳地100 mM至300 mM，更佳地150 mM至250 mM，最佳地180 mM至230 mM，具體而言，可約為210 mM。

此外，在根據本發明之醫藥組合物中，作為穩定劑使用之胺基酸可為1 mM至100 mM之範圍，較佳地3 mM至30 mM，更佳地5 mM至25 mM，最佳地7 mM至20 mM，具體而言，可為8 mM至15 mM之範圍。

根據本發明之醫藥組合物可更包含界面活性劑。

較佳地，界面活性劑可為非離子界面活性劑，例如聚氧乙烯-山梨醇脂肪酸酯(聚山梨醇酯或Tween)、聚乙烯-聚丙二醇、聚氧乙烯-硬脂酸酯、聚氧乙烯烷基醚(例如聚氧乙烯單月桂基醚、烷基苯基聚氧乙烯醚[Triton-X]及聚氧乙烯-聚氧丙烯共聚物[Poloxamer and Pluronic])以及十二烷基磺酸鈉(SDS)，但不限於此。

更佳地，可使用聚山梨醇酯。聚山梨醇酯可為聚山梨醇酯20或聚山梨醇酯80，但不限於此。

在根據本發明之醫藥組合物中，非離子界面活性劑的濃度可為0.0000001 %(w/v)(重量/體積)至0.5%(w/v)之範圍，較佳地0.000001 %(w/v)至0.4 %(w/v)，更佳地0.00001 %(w/v)至0.3 %(w/v)，最佳地0.001 %(w/v)至0.2 %(w/v)。

在一實施例中，根據本發明之醫藥組合物可包含50-350毫克/毫升之抗體(例如抗HER2抗體或免疫檢查點抗體)、提供pH 5.5 ± 2.0之組胺酸緩衝液、10-400 mM α,α-海藻糖、1-50 mM甲硫胺酸及0.0000001 %(w/v)至0.5 %(w/v)之聚山梨醇酯。

在更具體實施例中，根據本發明之醫藥組合物可包含120毫克/毫升之抗HER2抗體或免疫檢查點抗體、提供pH 5.5 ± 2.0之20 mM胺酸緩衝液、210 mM α,α-海藻糖、10 mM甲硫胺酸及2,000 單位/毫升之PH20變異體，並可更包含0.005 %(w/v)至0.1 %(w/v)之聚山梨醇酯。

根據本發明之醫藥組合物可透過以下方式給藥：靜脈注射、皮下注射、肌內注射、腹膜內注射、內皮給藥、局部給藥、肺內給藥、直腸內給藥及類似者，皮下給藥較佳地透過皮下注射來進行，更佳地使用醫藥組合物作為用於皮下注射的注射製劑。

因此，本發明另一實施例提供一製劑，包含根據本發明之醫藥組合物，較佳地用於皮下注射的注射製劑。

用於皮下注射的注射製劑可提供為不須額外稀釋過程的準備注射形式(ready-to-inject form)，並可在盛裝於預填充的注射筒、玻璃安瓿或塑膠容器中而被提供。

本發明亦關於一種治療疾病的方法，其使用根據本發明之醫藥組合物或製劑。

可使用根據本發明之醫藥組合物或製劑來治療的疾病並不特別限定，只要係可使用與根據本發明之PH20變異體結合之藥物來治療的疾病，即無限制。

可使用根據本發明之醫藥組合物或製劑來治療的疾病可為癌症或自體免疫疾病，但不限於此。

可使用根據本發明之醫藥組合物或製劑來治療的癌症或惡性腫瘤(carcinoma)並不特別限定，包含實體癌症(solid cancer)及血癌(blood cancer)兩者。這些癌症的示例包含皮膚癌(例如黑色素瘤)、肝癌、肝細胞腫瘤(hepatocellular carcinoma)、胃癌、乳癌、肺癌、卵巢癌、支氣管癌、鼻咽癌、喉癌、胰臟癌、膀胱癌、大腸直腸癌(colorectal cancer)、大腸癌(colon cancer)、子宮頸癌、腦癌、前列腺癌、骨癌、甲狀腺癌、副甲狀腺癌、腎癌、食道癌、膽道癌、睪丸癌、直腸癌(rectal cancer)、頭頸癌、子宮頸癌、輸尿管癌、骨肉瘤、神經母細胞瘤、纖維肉瘤、橫紋肌肉瘤、星狀細胞瘤、神經母細胞瘤及神經膠瘤，但不限於此。較佳地，可使用根據本發明之醫藥組合物或製劑來治療的癌症可選自由胃癌、大腸直腸癌、乳癌、肺癌及腎癌組成的群組，但不限於此。

可使用根據本發明之醫藥組合物或製劑來治療的自體免疫疾病包含類風溼性關節炎、哮喘、牛皮癬、多發性硬化症、過敏性鼻炎、克隆氏疾病(Crohn’s disease)、潰瘍性結腸炎、全身性紅斑狼瘡、第一型糖尿病、發炎性腸道疾病(inflammatory bowel disease，IBD)以及異位性皮炎，但不限於此。

本發明亦提供一種治療疾病的方法，包含將根據本發明之醫藥組合物或製劑給予需要治療的受試者，本發明更提供一種根據本發明之醫藥組合物或製劑用於治療疾病的用途。

除非另有定義，否則本發明中所使用之技術及科學術語具有如本發明所屬領域具有通常知識者所通常理解般的意義。此外，與習知技術中的那些相同的技術配置及操作的重複說明將會被省略。

在下文中，將參考示例進一步詳細說明本發明。這些示例僅提供用於說明目的，對於本領域中具有通常知識者而言顯而易見的是，這些示例不應被解釋為限制本發明的範圍。

〔示例〕

〔示例1，製劑研發〕

如表6所示，製備四種trastuzumab皮下注射製劑。製劑1至製劑4通常包含120毫克/毫升之trastuzumab，並由20 mM組胺酸/組胺酸-HCl (pH 5.5)、210 mM海藻糖、10 mM甲硫胺酸及PH20變異體組成。製劑1至製劑4之中的差異在於非離子界面活性劑的濃度，其中製劑1：0%聚山梨醇酯20，製劑2：0.005%聚山梨醇酯20，製劑3：0.04%聚山梨醇酯20，製劑4：0.1%聚山梨醇酯20。

表6，製劑的組成

	製劑1	製劑2	製劑3	製劑4
抗體	Trastuzumab (120毫克/毫升)
緩衝液	20 mM組胺酸/組胺酸-HCl
穩定劑1	210 mM海藻糖
穩定劑2	10 mM甲硫胺酸
聚山梨醇酯20	0%	0.005%	0.04%	0.1%
玻尿酸酶	SEQ ID NO: 44之HP46 (2,000單位/毫升)

〔示例2，使用分光光度計的量測〕

將製劑1至製劑4在45°C下放置14天，使用Beckman製造的分光光度計來分析蛋白質濃度的變化。各個樣品以蒸餾水稀釋，使得樣品的濃度為0.4毫克/毫升，接著使用分光光度計量測蛋白質在280奈米的吸收值。在嚴苛條件下(即在45°C下14天)的穩定性測試中，製劑1至製劑4的蛋白質濃度沒有顯著的變化。然而，玻尿酸酶的活性在45°C下急遽降低，因此在本示例中，沒有量測酵素活性(請見圖6)。

〔示例3，使用粒徑篩析層析研究各個製劑中Trastuzumab的單體比例〕

關於粒徑篩析層析分析(size-exclusion chromatography analysis)，使用Shimadzu Prominence提供之HPLC系統及TSK-gel G3000SWXL (7.8 X 300毫米，5微米)及TSK保護管柱(TSK guard column，6.0 x 4.0 毫米，7微米)。使用包含0.25 M氯化鉀的0.2 M磷酸鉀(pH 6.2)作為移動相。藉由應用等度分離模式(isocratic separation mode)以0.5毫升/分鐘之流速進行分析35分鐘。樣品以分析溶劑稀釋，使得最終濃度為10毫克/毫升，在將20微升之樣品注射至HPLC管柱之後，記錄管柱析出液在280奈米的吸收值。計算HPLC層析圖中Trastuzumab的單體比例並作圖。

當在嚴苛的條件下(即在45°C下14天)之穩定性測試中進行粒徑篩析層析分析時，製劑1至製劑4展現出相似的變化模式。主在變化在於高分子量(HMW)及低分子量(LMW)的降解產物增加，以及單體含量降低(約1.5%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在45°C)之穩定性測試中進行粒徑篩析層析分析的結果是，依據聚山梨醇酯20的濃度(0-0.1 %(w/v))，製劑之間的穩定性並沒有顯著的差異(請見圖1A~圖1B)。

〔示例4，量測包含Trastuzumab及HP46之製劑的蛋白質聚集溫度〕

使用動態光散射(Dynamic light scattering，DLS)分析因熱而造成之蛋白質的變性性質(denaturation property)。在本實驗中，量測依據溫度變化之蛋白質分子的尺寸變化，並將其用於計算蛋白質聚集溫度。關於動態光散射分析，使用Malvern提供之Zetasizer-nano-ZS儀器及石英光析槽(quartz cuvette，ZEN2112)。在分析過程中，溫度以1°C之間隔從25°C升高至85°C，使用各個製劑緩衝液將樣品稀釋至1毫克/毫升，接著將150µL之樣品加入光析槽中用於分析。

不包含聚山梨醇酯20的製劑1的聚集溫度為74°C，製劑2至製劑4的聚集溫度為76°C(請見圖2A及圖2B)。

〔示例5，包含Trastuzumab及HP46之製劑的弱陽離子交換層析量測〕

關於弱陽離子交換層析分析(WCX chromatography analysis)，使用Shimadzu Prominence提供之HPLC系統並使用TSKgel CM-STAT管柱(TSKgel CM-STAT column，4.6 x 100毫米，7微米)、TSKgel 保護凝膠CMSTAT (TSKgel guard gel CMSTAT，內徑3.2毫米x 1.5公分)及類似者作為管柱。移動相A為10 mM磷酸鈉(pH 7.5)，移動相B為包含0.1 M氯化鈉的10 mM磷酸鈉(pH 7.2)。以0.8毫升/分鐘之流速以0-30%移動相B之線性濃度梯度進行分析55分鐘。使用移動相A稀釋樣品，使得最終濃度為1.0毫克/毫升，將80微升之樣品注射至HPLC，接著記錄管柱析出液在280奈米的吸收值。計算HPLC層析圖中trastuzumab的單體比例並作圖。

當在嚴苛的條件下(即在45°C下14天)之穩定性測試中進行弱陽離子交換層析分析時，製劑1至製劑4展現出相似的變化模式。具體變化包含酸性變異體的相對含量增加(14天變化約30%)、主峰相對含量減少(14天變化約44%)以及鹼性變異體的相對含量增加(14天變化約15%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在45°C)之穩定性測試中的弱陽離子交換層析分析中，依據聚山梨醇酯20(0-0.1%)之蛋白質穩定性為相似的(請見圖3A~圖3D)。

〔示例6，製劑研發〕

如表7所示，製備三種trastuzumab皮下注射製劑。製劑5至製劑7通常包含120毫克/毫升之trastuzumab、20 mM組胺酸/組胺酸-HCl (pH 5.5)、210 mM海藻糖、10 mM甲硫胺酸及HP46。製劑5至製劑7之中的差異在於穩定劑3的成分，其中製劑5：0.04%聚山梨醇酯20，製劑6：50 mM Lys-Lys，製劑7：甘胺酸。

表7，製劑的組成

	製劑5	製劑6	製劑7
抗體	Trastuzumab (120毫克/毫升)
緩衝液	20 mM組胺酸/組胺酸-HCl
穩定劑 1	210 mM海藻糖
穩定劑 2	10 mM甲硫胺酸
穩定劑 3	0.04% 聚山梨醇酯20	50 mM Lys-Lys	50 mM甘胺酸
玻尿酸酶	SEQ ID NO: 44之HP46 (2,000單元/毫升)

〔示例7，使用分光光度計量測〕

將製劑5至製劑7在45°C下放置14天，使用Beckman製造的分光光度計來分析蛋白質濃度的變化。各個樣品以蒸餾水稀釋，使得樣品的濃度為0.4毫克/毫升，接著使用分光光度計量測蛋白質在280奈米的吸收值。在嚴苛條件下(即在45°C下14天)的穩定性測試中，製劑5至製劑7的蛋白質濃度沒有顯著的變化。然而，玻尿酸酶的活性在45°C下急遽降低，因此在本示例中，沒有量測酵素活性(請見圖6)。

〔示例8，使用粒徑篩析層析研究各個製劑中Trastuzumab的單體比例〕

關於粒徑篩析層析分析，使用Shimadzu Prominence提供之HPLC系統並使用TSK-gel G3000SWXL (7.8 X 300毫米，5微米)及TSK保護管柱(6.0 x 4.0毫米，7微米)作為管柱。使用包含0.25 M氯化鉀的0.2 M磷酸鉀(pH 6.2)作為移動相。以0.5毫升/分鐘之流速應用等度分離模式(isocratic separation mode)進行35分鐘。樣品以分析溶劑稀釋，使得最終濃度為10毫克/毫升，在將20微升之樣品注射至HPLC管柱之後，量測在280奈米的吸收值。計算HPLC層析圖中trastuzumab的單體比例並作圖。

當在嚴苛的條件下(即在45°C下14天)之穩定性測試中進行粒徑篩析層析分析時，製劑5至製劑7展現出相似的變化模式。主在變化在於高分子量(HMW)及低分子量(LMW)的雜質增加，以及單體含量降低(約1.5%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在45°C)之穩定性測試中進行粒徑篩析層析分析的結果是，在0.04%聚山梨醇酯20、50 mM Lys-Lys及50 mM甘胺酸的製劑中展現出相似的蛋白質穩定性(請見圖4)。

〔示例9，包含Trastuzumab及HP46之製劑的弱陽離子交換層析分析〕

關於弱陽離子交換層析分析，使用Shimadzu Prominence提供之HPLC系統並使用TSKgel CM-STAT (4.6 x 100毫米，7微米)、TSKgel保護凝膠CMSTAT (內徑3.2毫米x 1.5公分)及類似者作為管柱。移動相A為10 mM磷酸鈉(pH 7.5)，移動相B為包含0.1 M氯化鈉的10 mM磷酸鈉(pH 7.2)。藉由以0.8毫升/分鐘之流速使用0-30%移動相B之線性濃度梯度的分離模式進行分析55分鐘。使用移動相A稀釋樣品，使得最終濃度為1.0毫克/毫升，將80微升之樣品注射至HPLC，接著記錄在280奈米的吸收值。計算HPLC層析圖中trastuzumab的單體比例並作圖。

當在嚴苛的條件下(即在45°C下14天)之穩定性測試中進行弱陽離子交換層析分析時，製劑5至製劑7展現出相似的變化模式。具體變化包含酸性變異體的相對含量增加(14天變化約30%)、主峰相對含量減少(14天變化約44%)以及鹼性變異體的相對含量增加(14天變化約15%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在45°C)之穩定性測試中進行弱陽離子交換層析分析的結果是，在0.04%聚山梨醇酯20、50 mM Lys-Lys及50 mM甘胺酸的製劑中展現出相似的蛋白質穩定性(請見圖5A~圖5C)。

〔示例10，在Trastuzumab及HP46的皮下注射製劑中依據溫度40°C及45°C之HP46的穩定性評估〕

為了評估在trastuzumab的皮下注射製劑中HP46的穩定性，將trastuzumab (120 毫克/毫升)與PH20 (2000單位/毫升)混合。此時，所使用之緩衝液包含20 mM組胺酸(pH 5.5)、210 mM海藻糖、10 mM甲硫胺酸及0.04%聚山梨醇酯20。控制組樣品的酵素活性在第0天被量測，而實驗組樣品在40°C或45°C下放置1天，接著量測各個樣品的酵素活性。

將各個賀癌平皮下注射製劑(trastuzumab + HW2及trastuzumab + HP46)在40°C下放置1天，接著量測玻尿酸酶的活性，其結果，各情況分別展現出51%、47%及94%之活性，這表示HP46在40°C具有最佳的熱穩定性(請見圖6A~圖6B)。此外，將賀癌平皮下注射製劑(trastuzumab + HW2及trastuzumab + HP46)在45°C下放置1天，接著量測玻尿酸酶的活性，其結果，賀癌平皮下注射製劑及trastuzumab + HW2不具有酵素活性，但trastuzumab + HP46的酵素活性仍存在(請見圖6~圖6B)。

〔示例11，製劑研發〕

如表8所示，製備三種trastuzumab皮下注射製劑。製劑8至製劑10通常包含120毫克/毫升之trastuzumab、20 mM組胺酸/組胺酸-HCl (pH 5.5)、210 mM海藻糖、10 mM甲硫胺酸及PH20變異體。製劑8至製劑10之中的差異在於非離子界面活性劑的濃度，其中製劑8：0%聚山梨醇酯20，製劑9：0.005%聚山梨醇酯20，製劑10：0.04%聚山梨醇酯20。

表8，製劑的組成

	製劑8	製劑9	製劑10
抗體	Trastuzumab (120毫克/毫升)
聚山梨醇酯20	0%	0.005%	0.04%
緩衝液	20 mM組胺酸/組胺酸-HCl
穩定劑 1	210 mM海藻糖
穩定劑 2	10 mM甲硫胺酸
pH	5.5
玻尿酸酶	SEQ ID NO: 44之HP46 (2,000單位/毫升)

〔示例12，使用分光光度計的量測〕

將製劑8至製劑10在40°C下放置14天，使用Beckman製造的分光光度計來分析蛋白質濃度的變化。各個樣品以蒸餾水稀釋，使得樣品的濃度為0.4毫克/毫升，接著使用分光光度計量測蛋白質在280奈米的吸收值。在嚴苛條件下(即在40°C下14天)的穩定性測試中，製劑8至製劑10的蛋白質濃度沒有顯著的變化。

〔示例13，使用粒徑篩析層析研究各個製劑中Trastuzumab的單體比例〕

關於粒徑篩析層析分析，使用Shimadzu Prominence提供之HPLC系統並使用TSK-gel G3000SWXL (7.8 X 300毫米，5微米)及TSK保護管柱(6.0 x 4.0毫米，7微米)作為管柱。使用包含0.25 M氯化鉀的0.2 M磷酸鉀(pH 6.2)作為移動相。藉由應用等度分離模式(isocratic separation mode)以0.5毫升/分鐘之流速進行分析35分鐘。樣品以分析溶劑稀釋，使得最終濃度為10毫克/毫升，在將20微升之樣品注射至HPLC管柱之後，量測在280奈米的吸收值。計算HPLC層析圖中Trastuzumab的單體比例並作圖。

當在嚴苛的條件下(即在40°C下14天)之穩定性測試中進行粒徑篩析層析分析時，製劑8至製劑10展現出相似的變化模式。主在變化在於高分子量(HMW)及低分子量(LMW)的降解產物增加，以及單體含量降低(約低於1.0%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在40°C)之穩定性測試中進行粒徑篩析層析分析的結果是，依據聚山梨醇酯20的濃度(0-0.04%)，製劑之間的穩定性並沒有顯著的差異(請見圖7)。

〔示例14，量測包含Trastuzumab及HP46之製劑的蛋白質聚集溫度〕

在蛋白質藥物領域中，使用動態光散射(Dynamic light scattering，DLS)分析因熱而造成之蛋白質的變性性質(denaturation property)。在本實驗中，量測依據溫度變化之蛋白質分子的尺寸變化，並將其用於計算蛋白質聚集溫度。關於動態光散射分析，使用Malvern提供之Zetasizer-nano-ZS儀器及石英光析槽(ZEN2112)。在分析過程中，溫度以1°C之間隔從25°C升高至85°C，使用各個製劑緩衝液將樣品稀釋至1毫克/毫升，接著將150µL之樣品加入光析槽中用於分析。

不包含聚山梨醇酯20的製劑8的聚集溫度為78.3°C，製劑9展現出77.3°C之聚集溫度，製劑10展現出77.7°C之聚集溫度。在示例13中，即使不包含聚山梨醇酯20，蛋白質的單體比例也沒有出現變化，比較包含聚山梨醇酯20的情況與不包含聚山梨醇酯20的情況之結果，證實這些蛋白質之間的聚集沒有差異。這些結果表示對於trastuzumab的皮下注射製劑，不需要使聚山梨醇酯20的量最小化(請見圖8A~圖8B)。

〔示例15，包含Trastuzumab及HP46之製劑的弱陽離子交換層析分析〕

關於弱陽離子交換層析分析，使用Shimadzu Prominence提供之HPLC系統並使用TSKgel CM-STAT管柱(4.6 x 100毫米，7微米)、TSKgel保護凝膠CMSTAT (內徑3.2毫米 x 1.5公分)及類似者作為管柱。移動相A為10 mM磷酸鈉(pH 7.5)，移動相B為包含0.1 M氯化鈉的10 mM磷酸鈉(pH 7.2)。以0.8毫升/分鐘之流速以0-30%移動相B之線性濃度梯度進行分析55分鐘。使用移動相A稀釋樣品，使得最終濃度為1.0毫克/毫升，將80微升之樣品注射至HPLC，接著記錄管柱析出液在280奈米的吸收值。計算HPLC層析圖中trastuzumab的單體比例並作圖。

當在嚴苛的條件下(即在40°C下14天)之穩定性測試中進行弱陽離子交換層析分析時，製劑8至製劑10展現出相似的變化模式。具體變化包含酸性變異體的相對含量增加(14天變化約10%)、主峰相對含量減少(14天變化約40%)以及鹼性變異體的相對含量增加(14天變化約300%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在40°C)之穩定性測試中的弱陽離子交換層析分析中，依據聚山梨醇酯20(0-0.04%)之蛋白質穩定性為相似的(請見圖9A~圖9D)。

〔示例16，量測包含Trastuzumab及HP46之製劑的酵素活性〕

用於量測酵素活性的濁度法(Turbidimetric assay)係一種方法，其藉由吸收值來測量由殘留的玻尿酸與酸化的白蛋白(BSA)結合而形成之聚集的程度，當玻尿酸被PH20水解時，結合至白蛋白的量降低，造成吸收值降低。將作為標準品的BTH (Sigma)稀釋成1單位/毫升、2單位/毫升、5單位/毫升、7.5單位/毫升、10單位/毫升、15單位/毫升、20單位/毫升、30單位/毫升、50單位/毫升及60單位/毫升，並配製於各個管(tube)中。經純化的PH20變異體樣品以酵素稀釋緩衝液(20 mM Tris-HCl，pH 7.0，77 mM氯化鈉，0.01%(w/v)牛血清白蛋白)稀釋成100X、300X、600X、1200X及2400X，並配製於各管中。在乾淨的管中，將具有3毫克/毫升之濃度的玻尿酸酶溶液稀釋10倍變成0.3毫克/毫升之濃度，使得各管的體積變成180微升。將60微升之包含玻尿酸酶的樣品加入至經稀釋的玻尿酸溶液並與其混合，並在37°C下反應45分鐘。在反應完成之後，將50微升之經反應的酵素與250微升之酸性白蛋白溶液加入至96孔盤的各個孔中並搖晃10分鐘，接著使用分光光度計量測在600奈米的吸收值。

在嚴苛的條件下(即在40°C下14天)之穩定性測試中進行活性分析的結果是，證實聚山梨醇酯20的濃度愈高，活性隨時間降低愈多(請見圖10)。

〔示例17，製劑研發〕

如表9所示，製備三種trastuzumab皮下注射製劑。製劑11至製劑13通常包含120毫克/毫升之trastuzumab、20 mM組胺酸/組胺酸-HCl (pH 5.5)、210 mM海藻糖、10 mM甲硫胺酸及PH20變異體。製劑11至製劑13之中的差異在於非離子界面活性劑的濃度，其中製劑11：0%聚山梨醇酯80，製劑12：0.005%聚山梨醇酯80，製劑13：0.04%聚山梨醇酯80。

表9，製劑的組成

	製劑11	製劑12	製劑13
抗體	Trastuzumab (120 毫克/毫升)
聚山梨醇酯80	0%	0.005%	0.04%
緩衝液	20 mM組胺酸/組胺酸-HCl
穩定劑 1	210 mM海藻糖
穩定劑 2	10 mM甲硫胺酸
pH	5.5
玻尿酸酶	SEQ ID NO: 44之HP46 (2,000單位/毫升)

當在嚴苛的條件下(即在40°C下14天)之穩定性測試中進行粒徑篩析層析分析時，製劑11至製劑13展現出相似的變化模式。主在變化在於高分子量(HMW)及低分子量(LMW)的降解產物增加，以及單體含量降低(約低於1.0%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在40°C)之穩定性測試中進行粒徑篩析層析分析的結果是，依據聚山梨醇酯80的濃度(0-0.04%)，製劑之間的穩定性並沒有顯著的差異(請見圖11)。

〔示例18，包含Trastuzumab及HP46之製劑的弱陽離子交換層析分析〕

關於弱陽離子交換層析分析，使用Shimadzu Prominence提供之HPLC系統並使用TSKgel CM-STAT管柱 (4.6 x 100毫米，7微米)、TSKgel保護凝膠CMSTAT (內徑3.2毫米x 1.5公分)及類似者作為管柱。移動相A為10 mM磷酸鈉(pH 7.5)，移動相B為包含0.1 M氯化鈉的10 mM磷酸鈉(pH 7.2)。以0.8毫升/分鐘之流速以0-30%移動相B之線性濃度梯度進行分析55分鐘。使用移動相A稀釋樣品，使得最終濃度為1.0毫克/毫升，將80微升之樣品注射至HPLC，接著記錄管柱析出液在280奈米的吸收值。計算HPLC層析圖中trastuzumab的單體比例並作圖。

當在嚴苛的條件下(即在40°C下14天)之穩定性測試中進行弱陽離子交換層析分析時，製劑11至製劑13展現出相似的變化模式。具體變化包含酸性變異體的相對含量增加(14天變化約10%)、主峰相對含量減少(14天變化約40%)以及鹼性變異體的相對含量增加(14天變化約300%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在40°C)之穩定性測試中的弱陽離子交換層析分析中，依據聚山梨醇酯80(0-0.04%)之蛋白質穩定性為相似的(請見圖12A~圖12D)。

〔示例19，量測包含Trastuzumab及HP46之製劑的酵素活性〕

用於量測酵素活性的濁度法(Turbidimetric assay)係一種方法，其藉由吸收值來測量由殘留的玻尿酸與酸化的白蛋白(BSA)結合而形成之聚集的程度，當玻尿酸被PH20水解時，結合至白蛋白的量降低，造成吸收值降低。將作為標準品的BTH (Sigma)稀釋成1單位/毫升、2單位/毫升、5單位/毫升、7.5單位/毫升、10單位/毫升、15單位/毫升、20單位/毫升、30單位/毫升、50單位/毫升及60單位/毫升，並配製於各個管(tube)中。經純化的蛋白質樣品以酵素稀釋緩衝液(20 mM Tris-HCl，pH 7.0，77 mM氯化鈉，0.01%(w/v)牛血清白蛋白)稀釋成100X、300X、600X、1200X及2400X，並配製於各管中。在乾淨的管中，將具有3毫克/毫升之濃度的玻尿酸酶溶液稀釋10倍變成0.3毫克/毫升之濃度，使得各管的體積變成180微升。將60微升之包含玻尿酸酶的樣品加入至經稀釋的玻尿酸溶液並與其混合，並在37°C下反應45分鐘。在反應完成之後，將50微升之經反應的酵素與250微升之酸性白蛋白溶液加入至96孔盤的各個孔中並搖晃10分鐘，接著使用分光光度計量測在600奈米的吸收值。

在嚴苛的條件下(即在40°C下14天)之穩定性測試中進行活性分析的結果是，證實聚山梨醇酯80的濃度愈高，活性隨時間降低愈多(請見圖13)。

〔示例20，製劑研發〕

如表10所示，製備三種rituximab製劑。製劑14至製劑16通常包含120毫克/毫升之rituximab、20 mM組胺酸/組胺酸-HCl (pH 5.5)、210 mM海藻糖、10 mM甲硫胺酸及PH20變異體。製劑14至製劑16之中的差異在於非離子界面活性劑的濃度，其中製劑14：0%聚山梨醇酯80，製劑15：0.005%聚山梨醇酯80，製劑16：0.06%聚山梨醇酯80。

表10，製劑的組成s

	製劑14	製劑15	製劑16
Rituximab	120 毫克/毫升 (± 10)
PS 80	0%	0.005%	0.06%
緩衝液	20 mM組胺酸/組胺酸-HCl
穩定劑 1	210 mM海藻糖
穩定劑 2	10 mM甲硫胺酸
pH	5.5
玻尿酸酶	SEQ ID NO: 44之HP46 (2,000單位/毫升)

當在嚴苛的條件下(即在40°C下7天)之穩定性測試中進行粒徑篩析層析分析時，製劑14至製劑16展現出相似的變化模式。主在變化在於高分子量(HMW)及低分子量(LMW)的降解產物增加，以及單體含量降低(約低於1.0%)，並且不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在40°C)之穩定性測試中進行粒徑篩析層析分析的結果是，依據聚山梨醇酯80的濃度(0-0.06%)，製劑之間的穩定性並沒有顯著的差異(請見圖14)。

〔示例21，量測包含Rituximab及HP46之製劑的酵素活性〕

用於量測酵素活性的濁度法(Turbidimetric assay)係一種方法，其藉由吸收值來測量由殘留的玻尿酸與酸化的白蛋白(BSA)結合而形成之聚集的程度，當玻尿酸被PH20水解時，結合至白蛋白的量降低，造成吸收值降低。將作為標準品的BTH (Sigma)稀釋成1單位/毫升、2單位/毫升、5單位/毫升、7.5單位/毫升、10單位/毫升、15單位/毫升、20單位/毫升、30單位/毫升、50單位/毫升及60單位/毫升，並配製於各個管(tube)中。經純化的蛋白質樣品以酵素稀釋緩衝液(20 mM Tris-HCl，pH 7.0，77 mM氯化鈉，0.01%(w/v)牛血清白蛋白)稀釋成100X、300X、600X、1200X及2400X，並配製於各管中。在乾淨的管中，將具有3毫克/毫升之濃度的玻尿酸酶溶液稀釋10倍變成0.3毫克/毫升之濃度，使得各管的體積變成180微升。將60微升之包含玻尿酸酶的樣品加入至經稀釋的玻尿酸溶液並與其混合，並在37°C下反應45分鐘。在反應完成之後，將50微升之經反應的酵素與250微升之酸性白蛋白溶液加入至96孔盤的各個孔中並搖晃10分鐘，接著使用分光光度計量測在600奈米的吸收值。

在嚴苛的條件下(即在40°C下7天)之穩定性測試中進行活性分析的結果是，證實聚山梨醇酯80的濃度愈高，活性隨時間降低愈多(請見圖15)。

〔示例22，量測不包含聚山梨醇酯之市售可得之製劑的酵素活性〕

如表11所示，製備兩種市售可得之rituximab製劑。製劑17係用於皮下注射製劑之市售可得的緩衝液，製劑18係用於靜脈注射製劑之市售可得的緩衝液。製劑17及製劑18包含PH20變異體且分別包含120毫克/毫升之rituximab及100毫克/毫升之rituximab，但不同於市售可得之產品而不包含聚山梨醇酯80。

表11，製劑的組成

	製劑17	製劑18
Rituximab	120 毫克/毫升	100 毫克/毫升
緩衝液	20 mM 組胺酸/組胺酸-HCl	25 mM檸檬酸鈉
穩定劑 1	210 mM海藻糖	145 mM氯化鈉
穩定劑 2	10 mM甲硫胺酸	10 mM甲硫胺酸
pH	5.5	6.5
玻尿酸酶	SEQ ID NO: 44之HP46 (2,000單位/毫升)

用於量測酵素活性的濁度法係一種方法，其藉由吸收值來測量由殘留的玻尿酸與酸化的白蛋白(BSA)結合而形成之聚集的程度，當玻尿酸被PH20水解時，結合至白蛋白的量降低，造成吸收值降低。將作為標準品的BTH (Sigma)稀釋成1單位/毫升、2單位/毫升、5單位/毫升、7.5單位/毫升、10單位/毫升、15單位/毫升、20單位/毫升、30單位/毫升、50單位/毫升及60單位/毫升，並配製於各個管中。經純化的蛋白質樣品以酵素稀釋緩衝液(20 mM Tris·HCl，pH 7.0，77 mM氯化鈉，0.01%(w/v)牛血清白蛋白)稀釋成100X、300X、600X、1200X及2400X，並配製於各管中。在乾淨的管中，將具有3毫克/毫升之濃度的玻尿酸酶溶液稀釋10倍變成0.3毫克/毫升之濃度，使得各管的體積變成180微升。將60微升之包含玻尿酸酶的樣品加入至經稀釋的玻尿酸溶液並與其混合，並在37°C下反應45分鐘。在反應完成之後，將50微升之經反應的酵素與250微升之酸性白蛋白溶液加入至96孔盤的各個孔中並搖晃10分鐘，接著使用分光光度計量測在600奈米的吸收值。

在嚴苛的條件下(即在40°C下6天)之穩定性測試中進行活性分析的結果是，證實即使在製劑中不包含聚山梨醇酯80，仍可維持高活性，尤其，製劑18維持高活性(請見圖16)。

〔示例23：製劑研發〕

如表12所示，製備四種pembrolizumab製劑。製劑19、製劑20及製劑21通常包含25毫克/毫升之pembrolizumab、10 mM組胺酸(pH 5.5)、7%蔗糖、10 mM甲硫胺酸及PH20變異體。製劑19至製劑21之中的差異在於非離子界面活性劑的濃度，其中製劑19：0%聚山梨醇酯80，製劑20：0.005%聚山梨醇酯80，製劑21：0.02%聚山梨醇酯80。製劑22包含25毫克/毫升之pembrolizumab並由10 mM組胺酸(pH 5.5)、210 mM海藻糖、10 mM甲硫胺酸、0.02%聚山梨醇酯80及PH20變異體組成。

表12，製劑的組成

	製劑19	製劑20	製劑21	製劑22
抗體	Pembrolizumab (25 毫克/毫升)
緩衝液	10 mM組胺酸 (pH 5.5)
穩定劑 1	7%蔗糖	7%蔗糖	7%蔗糖	210 mM 海藻糖
穩定劑 2	10 mM 甲硫胺酸	10 mM 甲硫胺酸	10 mM 甲硫胺酸	10 mM 甲硫胺酸
聚山梨醇酯80	0%	0.005%	0.02%	0.02%
玻尿酸酶	SEQ ID NO: 44之HP46 (2,000單位/毫升)

〔示例24，使用分光光度計的量測〕

將製劑19至製劑22在40°C下放置7天，使用Beckman製造的分光光度計來分析蛋白質濃度的變化。各個樣品以蒸餾水稀釋，使得樣品的濃度為0.4毫克/毫升，接著使用分光光度計量測蛋白質在280奈米的吸收值。

在嚴苛條件下(即在40°C下7天)的穩定性測試中，製劑19至製劑22的蛋白質濃度沒有顯著的變化。

〔示例25，使用粒徑篩析層析研究各個製劑中Pembrolizumab的單體比例〕

關於粒徑篩析層析分析，使用Shimadzu Prominence提供之HPLC系統並使用TSK-gel G3000SWXL (7.8 X 300毫米，5微米)及TSK保護管柱(6.0 x 4.0毫米，7微米)作為管柱。使用包含0.25 M氯化鉀的0.2 M磷酸鉀(pH 6.2)作為移動相。藉由應用等度分離模式(isocratic separation mode)以0.5毫升/分鐘之流速進行分析35分鐘。樣品以分析溶劑稀釋，使得最終濃度為10毫克/毫升，在將20微升之樣品注射至HPLC管柱之後，量測在280奈米的吸收值。計算HPLC層析圖中pembrolizumab的單體比例並作圖。

當在嚴苛的條件下(即在40°C下7天)之穩定性測試中進行粒徑篩析層析分析時，製劑19至製劑22展現出相似的變化模式。在高分子量(HMW)及低分子量(LMW)的降解產物中不依據製劑而有顯著的差異。結論，在嚴苛的條件下(即在40°C)之穩定性測試中進行粒徑篩析層析分析的結果是，製劑19至製劑22並未展現出顯著的差異，且亦不依據糖的類型而有差異(請見圖17)。這些結果與根據先前示例之trastuzumab及rituximab的情況之結果一致。

〔示例26，量測包含Pembrolizumab及HP46之製劑的酵素活性〕

用於量測酵素活性的濁度法係一種方法，其藉由吸收值來測量由殘留的玻尿酸與酸化的白蛋白(BSA)結合而形成之聚集的程度，當玻尿酸被PH20水解時，結合至白蛋白的量降低，造成吸收值降低。將作為標準品的BTH (Sigma)稀釋成1單位/毫升、2單位/毫升、5單位/毫升、7.5單位/毫升、10單位/毫升、15單位/毫升、20單位/毫升、30單位/毫升、50單位/毫升及60單位/毫升，並配製於各個管(tube)中。經純化的蛋白質樣品以酵素稀釋緩衝液(20 mM Tris·HCl，pH 7.0，77 mM氯化鈉，0.01%(w/v)牛血清白蛋白)稀釋成100X、300X、600X、1200X及2400X，並配製於各管中。在乾淨的管中，將具有3毫克/毫升之濃度的玻尿酸酶溶液稀釋10倍變成0.3毫克/毫升之濃度，使得各管的體積變成180微升。將60微升之包含酵素的樣品加入至經稀釋的玻尿酸溶液並與其混合，並在37°C下反應45分鐘。在反應完成之後，將50微升之經反應的酵素與250微升之酸性白蛋白溶液加入至96孔盤的各個孔中並搖晃10分鐘，接著使用分光光度計量測在600奈米的吸收值。

在嚴苛的條件下(即在40°C下7天)之穩定性測試中進行活性分析的結果是，證實隨著聚山梨醇酯80的濃度增加，活性的下降大。亦證實，當包含相同量的聚山梨醇酯80時，在含海藻糖之製劑中降低的活性小於在含蔗糖之製劑中降低的活性(請見圖18)。

〔示例27，HP46及野生型HW2的pH-活性圖〕

關於用以確認HP46及野生型HW2的pH-活性圖的實驗，使用微量濁度分析法(microturbidimetric assay method)。對於各個pH值，製備用於溶解作為受質之玻尿酸的玻尿酸緩衝液及用於稀釋酵素的酵素緩衝液。

準備共三個96孔盤用於酵素與受質之間的反應，並將其標示為A、B及C，並進行實驗。

使用20 mM乙酸及70 mM氯化鈉來製備pH為4.0、4.5或5.0的玻尿酸緩衝液，使用20 mM磷酸鈉及70 mM氯化鈉來製備pH為5.5、6.0、6.5、7.0或8.0的玻尿酸溶液。將20毫克之玻尿酸溶解於10毫升之各個準備好的玻尿酸緩衝液中，以製備最終玻尿酸受質溶液，接著將其以依據pH製備之各個玻尿酸緩衝液稀釋，以製備成濃度為0.1毫克/毫升、0.25毫克/毫升、0.45毫克/毫升或0.7毫克/毫升的500微升之最終溶液，將100微升之各個溶液分配至標示為A之96孔盤的各個孔中。使用依據濃度製備並稀釋的玻尿酸緩衝液作為檢量曲線以量測玻尿酸的濃度。

使用20 mM乙酸、0.01 %(w/v) BSA及70 mM氯化鈉來製備pH為4.0、4.5或5.0的酵素緩衝液，使用20 mM磷酸鈉、0.01 %(w/v) BSA及70 mM氯化鈉來製備pH為5.5、6.0、6.5、7.0或8.0的酵素緩衝液。

HP46及野生型HW2酵素以依據pH值製備的酵素緩衝液配製成10單位/毫升，將50微升之最終溶液分配至標示為B之96孔盤的各個孔中。

將50微升之樣品從標示為A之96孔盤的各個孔中轉移至標示為B之96孔盤的各個孔中，接著在37°C反應，搖晃培養箱45分鐘。在反應完成前的15分鐘，將200微升之酸性白蛋白溶液分配於標示為C之96孔盤的各個孔中，當酵素受質反應完成時，將40微升之樣品從標示為B之96孔盤的各個孔中轉移至標示為C之96孔盤的各個孔中，接著反應20分鐘。在20分鐘後，量測在600奈米的吸收值，計算酵素受質反應之後殘留的玻尿酸的量，完成依據pH值的酵素活性圖(請見圖19)。

〔示例28，在史道二氏大鼠(Sprague-Dawley Rat)中使用賀癌平皮下注射製劑與Trastuzumab及HP46測試藥物動力學〕

為了測試trastuzumab及HP46的皮下注射製劑是否與賀癌平皮下注射製劑展現出相同的藥物動力學性質，使用9週齡之史道二氏大鼠進行實驗。賀癌平及trastuzumab的給藥劑量為18毫克/公斤(老鼠體重)，包含於賀癌平皮下注射製劑之rHuPH20的量為100 U，HP46的量亦為100 U。在藥物動力學的測試中，trastuzumab及HP46與賀癌平皮下注射製劑展現出相同的曲線下面積(Area Under the Curve，AUC)(請見圖20)。

根據本發明之醫藥組合物可用於皮下注射並且亦相當穩定，PH20變異體與藥物的活性可維持長時間，藥物較佳地為抗體藥物或類似物。因此，醫藥組合物不僅有助於降低生產皮下注射製劑的成本，亦可降低醫療成本，對患者的便利性而言相當有效。

儘管已為了說明而揭露本發明之較佳的實施例，但本領域具有通常知識者應理解，在不脫離如請求項所揭露之本發明的範圍及精神下，可做多種修改、添加及替代。

〔參考文獻〕 Bookbinder, L.H., Hofer, A., Haller, M.F., Zepeda, M.L., Keller, G.A., Lim, J.E., Edgington, T.S., Shepard, H.M., Patton, J.S., and Frost, G.I. (2006). A recombinant human enzyme for enhanced interstitial transport of therapeutics. J Control Release 114, 230-241. Borders jr., C.L. and Raftery, A. (1968) Purification and Partial Characterization of Testicular Hyaluronidase. J Biol Chem 243, 3756-3762 Chao, K.L., Muthukumar, L., and Herzberg, O. (2007). Structure of human hyaluronidase-1, a hyaluronan hydrolyzing enzyme involved in tumor growth and angiogenesis. Biochemistry 46, 6911-6920. Chen, K.J., Sabrina, S., El-Safory, N.S., Lee, G.C., and Lee, C.K. (2016) Constitutive expression of recombinant human hyaluronidase PH20 by Pichia pastoris. J Biosci Bioeng. 122, 673-678 Frost, G.I. (2007). Recombinant human hyaluronidase (rHuPH20): an enabling platform for subcutaneous drug and fluid administration. Expert Opin Drug Deliv 4, 427-440 Hofinger, E.S., Bernhardt, G., and Buschauer, A. (2007) Kinetics of Hyal-1 and PH-20 hyaluronidases: comparison of minimal substrates and analysis of the transglycosylation reaction. Glycobiology 17, 963-971 Kreidieh, F.Y., Moukadem, H.A., and Saghir, N.S.E. (2016) Overview, prevention and management of chemotherapy extravasation. World J Clin Oncol 7, 87-97. Thomas, J.R., Yocum, R.C., Haller, M.F., and Flament J. (2009) The INFUSE-Morphine IIB Study: Use of Recombinant Human Hyaluronidase (rHuPH20) to Enhance the Absorption of Subcutaneous Morphine in Healthy Volunteers. J Pain Symptom Manag 38, 673-682

無。

由以下詳細描述一併參考附圖將更清楚地理解本發明之上述及其他目的、特徵及其他優點。 圖1A示出在45°C之嚴苛的條件下之穩定性測試中trastuzumab的粒徑篩析層析圖。 圖1B示出在45°C之嚴苛的條件下之穩定性測試中依據製劑之trastuzumab單體蛋白的純度的變化。 圖2A及圖2B示出量測包含trastuzumab及新穎的PH20變異體HP46之製劑的蛋白質聚集溫度的結果。 圖3A為在45°C之嚴苛的條件下之穩定性測試中trastuzumab的弱陽離子交換(weak cation exchange，WCX)層析圖。 圖3B示出在45°C之嚴苛的條件下之穩定性測試中在製劑中之酸性變異體的相對量的變化(%)。 圖3C示出在45°C之嚴苛的條件下之穩定性測試中在製劑中之主峰的相對量的變化(%)。 圖3D示出45°C之嚴苛的條件下之穩定性測試中在製劑中之鹼性變異體的相對量的變化(%)。 圖4示出在45°C之嚴苛的條件下之穩定性測試中在製劑5-7中之trastuzumab單體蛋白的純度的變化。 圖5A示出在45°C之嚴苛的條件下之穩定性測試中在製劑5-7中之酸性變異體的相對量的變化(%)。 圖5B示出在45°C之嚴苛的條件下之穩定性測試中在製劑5-7中之主峰的相對量的變化(%)。 圖5C示出在45°C之嚴苛的條件下之穩定性測試中製劑5-7中之鹼性變異體的相對量的變化(%)。 圖6A示出在40°C之嚴苛的條件下之穩定性測試中量測在第0天及第1天之賀癌平(Herceptin)皮下注射製劑(賀癌平SC)、trastuzumab +野生型PH20 (HW2)及trastuzumab + PH20變異體HP46之殘餘的酵素活性的結果。 圖6B示出在在45°C之嚴苛的條件下之穩定性測試中量測在第0天及第1天之賀癌平皮下注射製劑、trastuzumab +野生型PH20 (HW2)及trastuzumab + PH20變異體HP46之殘餘的酵素活性的結果。 圖7示出在40°C之嚴苛的條件下14天之穩定性測試中製劑8-10的粒徑篩析層析分析結果。 圖8A示出使用動態光散射(Dynamic light scattering，DLS)設備量測蛋白質顆粒尺寸的變化的結果。 圖8B示出量測蛋白質聚集溫度的結果。 圖9A示出在40°C之嚴苛的條件下之穩定性測試中製劑8的弱陽離子交換(weak cation exchange，WCX)層析圖。 圖9B示出在40°C之嚴苛的條件下之穩定性測試中在製劑8-10中之酸性變異體的相對量的變化(%)。 圖9C示出在40°C之嚴苛的條件下之穩定性測試中在製劑8-10中之主峰的相對量的變化(%)。 圖9D示出在40°C之嚴苛的條件下之穩定性測試中在製劑8-10中之鹼性變異體的相對量的變化(%)。 圖10示出在40°C之嚴苛的條件下之穩定性測試中製劑8-10的相對酵素活性的變化(%)。 圖11示出在40°C之嚴苛的條件下之穩定性測試中製劑11-13之trastuzumab單體的純度的變化。 圖12A示出在40°C之嚴苛的條件下之穩定性測試中製劑11的弱陽離子交換(weak cation exchange，WCX)層析圖。 圖12B示出在40°C之嚴苛的條件下之穩定性測試中在製劑11-13中之酸性變異體的相對量的變化(%)。 圖12C示出在40°C之嚴苛的條件下之穩定性測試中在製劑11-13中之主峰的相對量的變化(%)。 圖12D示出在40°C之嚴苛的條件下之穩定性測試中在製劑11-13中之鹼性變異體的相對量的變化(%)。 圖13示出在40°C之嚴苛的條件下之穩定性測試中製劑11-13的相對酵素活性的變化(%)。 圖14示出在40°C之嚴苛的條件下之穩定性測試中製劑14-16中之rituximab單體的純度的變化。 圖15示出在40°C之嚴苛的條件下之穩定性測試中製劑14-16的相對酵素活性的變化(%)。 圖16示出在40°C之嚴苛的條件下之穩定性測試中製劑17及18的相對酵素活性的變化。 圖17示出在40°C之製劑19-22的粒徑篩析層析分析結果。 圖18示出在40°C之嚴苛的條件下之穩定性測試中製劑19-22的相對酵素活性的變化。 圖19示出重組人類PH20及HP46根據pH變化之酵素活性的變化。 圖20示出在9週齡之史道二氏大鼠(Sprague-Dawley rats)中賀癌平皮下注射產品(賀癌平 SC)及賀癌平皮下注射之生物相似候選者(trastuzumab + HP46; 賀癌平 SC BS)的藥物動力學的實驗結果，其中賀癌平及賀癌平生物相似候選者以每隻18毫克/公斤之量注射，皮下注射製劑包含100單位之rHuPH20及100單位之HP46(於pH 5.3)。

<110> 南韓商阿特根公司ALTEOGEN INC.

<120> 用於皮下注射之包含人類玻尿酸酶PH20變異體及藥物的醫藥組合物

<130> PP-B2376

<160> 51

<170> PatentIn version 3.5

<210> 1

<211> 509

<212> PRT

<213> 人工序列

<220>

<223> 合成序列

<400> 1

Gly Ile Pro Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys 115                 120                 125 Lys Asp Ile Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val 130                 135                 140 Ile Asp Trp Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro 145                 150                 155                 160 Lys Asp Val Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn 165                 170                 175 Val Gln Leu Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe 180                 185                 190 Glu Lys Ala Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys 195                 200                 205 Leu Leu Arg Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys 210                 215                 220 Tyr Asn His His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn 225                 230                 235                 240 Val Glu Ile Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser 245                 250                 255 Thr Ala Leu Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val 260                 265                 270 Ala Ala Thr Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val 275                 280                 285 Ser Lys Ile Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr 290                 295                 300 Arg Ile Val Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu 305                 310                 315                 320 Leu Val Tyr Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile 325                 330                 335 Val Ile Trp Gly Thr Leu Ser Ile Met Arg Ser Met Lys Ser Cys Leu 340                 345                 350 Leu Leu Asp Asn Tyr Met Glu Thr Ile Leu Asn Pro Tyr Ile Ile Asn 355                 360                 365 Val Thr Leu Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln 370                 375                 380 Gly Val Cys Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu 385                 390                 395                 400 Asn Pro Asp Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr 405                 410                 415 Val Arg Gly Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys 420                 425                 430 Phe Tyr Cys Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp 435                 440                 445 Val Lys Asp Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys 450                 455                 460 Ile Asp Ala Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile 465                 470                 475                 480 Phe Tyr Asn Ala Ser Pro Ser Thr Leu Ser Ala Thr Met Phe Ile Val 485                 490                 495 Ser Ile Leu Phe Leu Ile Ile Ser Ser Val Ala Ser Leu 500                 505 ＜210＞    2 ＜211＞    26 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    2 Met Ala Thr Gly Ser Arg Thr Ser Leu Leu Leu Ala Phe Gly Leu Leu 1               5                  10                  15 Cys Leu Pro Trp Leu Gln Glu Gly Ser Ala 20                  25 ＜210＞    3 ＜211＞    18 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    3 Met Lys Trp Val Thr Phe Ile Ser Leu Leu Phe Leu Phe Ser Ser Ala 1               5                  10                  15 Tyr Ser ＜210＞    4 ＜211＞    21 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    4 Met Ala Ala His Leu Leu Pro Ile Cys Ala Leu Phe Leu Thr Leu Leu 1               5                  10                  15 Asp Met Ala Gln Gly 20 ＜210＞    5 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    5 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Gly Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    6 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    6 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Phe Ile Leu Asn Val Thr Ser 325                 330                 335 Gly Ala Leu Leu Cys Ser Gln Ala Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    7 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    7 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Gly Pro Phe Ile Leu Asn Val Thr Ser 325                 330                 335 Gly Ala Leu Leu Cys Ser Gln Ala Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    8 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    8 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Val Ser Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Gly Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    9 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    9 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    10 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    10 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Val Ser Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    11 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    11 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    12 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    12 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    13 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    13 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    14 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    14 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Phe Ile Leu Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    15 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    15 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Phe Ile Leu Asn Val Thr Ser 325                 330                 335 Gly Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    16 ＜211＞    453 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    16 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser 435                 440                 445 Pro Ser Thr Leu Ser 450 ＜210＞    17 ＜211＞    430 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    17 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile 420                 425                 430 ＜210＞    18 ＜211＞    433 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    18 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe ＜210＞    19 ＜211＞    436 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    19 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro 435 ＜210＞    20 ＜211＞    452 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    20 Phe Arg Gly Pro Leu Leu Pro Asn Arg Pro Phe Leu Trp Ala Trp Asn 1               5                  10                  15 Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met 20                  25                  30 Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln 35                  40                  45 Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile 50                  55                  60 Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile 65                  70                  75                  80 Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr 85                  90                  95 Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp 100                 105                 110 Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn 115                 120                 125 Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr 130                 135                 140 Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp 145                 150                 155                 160 Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His 165                 170                 175 Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys 180                 185                 190 Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn 195                 200                 205 Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser 210                 215                 220 Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val 225                 230                 235                 240 Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala 245                 250                 255 Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp 260                 265                 270 Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly 275                 280                 285 Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu 290                 295                 300 Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met 305                 310                 315                 320 Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys 325                 330                 335 Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys 340                 345                 350 Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala 355                 360                 365 Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr 370                 375                 380 Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr 385                 390                 395                 400 Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala 405                 410                 415 Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys 420                 425                 430 Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro 435                 440                 445 Ser Thr Leu Ser 450 ＜210＞    21 ＜211＞    452 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    21 Phe Arg Gly Pro Leu Leu Pro Asn Arg Pro Phe Thr Thr Val Trp Asn 1               5                  10                  15 Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met 20                  25                  30 Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln 35                  40                  45 Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile 50                  55                  60 Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile 65                  70                  75                  80 Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr 85                  90                  95 Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp 100                 105                 110 Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn 115                 120                 125 Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr 130                 135                 140 Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp 145                 150                 155                 160 Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His 165                 170                 175 Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys 180                 185                 190 Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn 195                 200                 205 Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser 210                 215                 220 Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val 225                 230                 235                 240 Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala 245                 250                 255 Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp 260                 265                 270 Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly 275                 280                 285 Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu 290                 295                 300 Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met 305                 310                 315                 320 Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys 325                 330                 335 Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys 340                 345                 350 Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala 355                 360                 365 Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr 370                 375                 380 Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr 385                 390                 395                 400 Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala 405                 410                 415 Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys 420                 425                 430 Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro 435                 440                 445 Ser Thr Leu Ser 450 ＜210＞    22 ＜211＞    433 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    22 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys ＜210＞    23 ＜211＞    431 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    23 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe 420                 425                 430 ＜210＞    24 ＜211＞    455 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    24 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Ser Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 Phe Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn 435                 440                 445 Ala Ser Pro Ser Thr Leu Ser 450                 455 ＜210＞    25 ＜211＞    451 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    25 Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp Asn Ala 1               5                  10                  15 Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met Ser 20                  25                  30 Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln Gly 35                  40                  45 Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile Asp 50                  55                  60 Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile Ser 65                  70                  75                  80 Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr Met 85                  90                  95 Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp Arg 100                 105                 110 Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn Arg 115                 120                 125 Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr Glu 130                 135                 140 Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp Phe 145                 150                 155                 160 Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His Leu 165                 170                 175 Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys Lys 180                 185                 190 Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn Asp 195                 200                 205 Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser Ile 210                 215                 220 Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val Arg 225                 230                 235                 240 Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala Lys 245                 250                 255 Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp Gln 260                 265                 270 Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly Glu 275                 280                 285 Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu Ser 290                 295                 300 Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met Asp 305                 310                 315                 320 Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys Met 325                 330                 335 Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys Asn 340                 345                 350 Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala Ile 355                 360                 365 Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr Leu 370                 375                 380 Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr Ser 385                 390                 395                 400 Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala Val 405                 410                 415 Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys Pro 420                 425                 430 Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro Ser 435                 440                 445 Thr Leu Ser 450 ＜210＞    26 ＜211＞    449 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    26 Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp Asn Ala Pro Ser 1               5                  10                  15 Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met Ser Leu Phe 20                  25                  30 Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln Gly Val Thr 35                  40                  45 Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile Asp Ser Ile 50                  55                  60 Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile Ser Leu Gln 65                  70                  75                  80 Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr Met Pro Val 85                  90                  95 Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp Arg Pro Thr 100                 105                 110 Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn Arg Ser Ile 115                 120                 125 Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr Glu Ala Thr 130                 135                 140 Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp Phe Leu Val 145                 150                 155                 160 Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His Leu Trp Gly 165                 170                 175 Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys Lys Pro Gly 180                 185                 190 Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn Asp Asp Leu 195                 200                 205 Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser Ile Tyr Leu 210                 215                 220 Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val Arg Asn Arg 225                 230                 235                 240 Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala Lys Ser Pro 245                 250                 255 Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp Gln Val Leu 260                 265                 270 Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly Glu Thr Val 275                 280                 285 Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu Ser Ile Thr 290                 295                 300 Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met Asp Thr Thr 305                 310                 315                 320 Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys Met Cys Ser 325                 330                 335 Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys Asn Trp Asn 340                 345                 350 Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala Ile Gln Leu 355                 360                 365 Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr Leu Glu Asp 370                 375                 380 Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr Ser Thr Leu 385                 390                 395                 400 Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala Val Asp Val 405                 410                 415 Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys Pro Pro Met 420                 425                 430 Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro Ser Thr Leu 435                 440                 445 Ser ＜210＞    27 ＜211＞    432 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    27 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 ＜210＞    28 ＜211＞    429 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    28 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys 420                 425 ＜210＞    29 ＜211＞    426 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    29 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys Ile Ala Asp 420                 425 ＜210＞    30 ＜211＞    423 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    30 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val Asp Val Cys 420 ＜210＞    31 ＜211＞    420 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    31 Leu Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp 1               5                  10                  15 Ala Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro 20                  25                  30 Leu Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala 35                  40                  45 Thr Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr 50                  55                  60 Pro Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro 65                  70                  75                  80 Gln Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile 85                  90                  95 Thr Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp 100                 105                 110 Glu Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val 115                 120                 125 Tyr Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu 130                 135                 140 Ser Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala 145                 150                 155                 160 Gly Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg 165                 170                 175 Pro Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His 180                 185                 190 His Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile 195                 200                 205 Lys Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu 210                 215                 220 Tyr Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr 225                 230                 235                 240 Leu Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile 245                 250                 255 Pro Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val 260                 265                 270 Phe Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr 275                 280                 285 Thr Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp 290                 295                 300 Gly Thr Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys 305                 310                 315                 320 Glu Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu 325                 330                 335 Ala Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys 340                 345                 350 Ile Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp 355                 360                 365 Asn Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly 370                 375                 380 Lys Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys 385                 390                 395                 400 Ser Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp 405                 410                 415 Thr Asp Ala Val 420 ＜210＞    32 ＜211＞    433 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    32 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys ＜210＞    33 ＜211＞    435 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    33 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro 435 ＜210＞    34 ＜211＞    436 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    34 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met 435 ＜210＞    35 ＜211＞    437 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    35 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu 435 ＜210＞    36 ＜211＞    438 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    36 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr 435 ＜210＞    37 ＜211＞    439 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    37 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr Glu 435 ＜210＞    38 ＜211＞    454 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    38 Asn Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala 1               5                  10                  15 Trp Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu 20                  25                  30 Asp Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr 35                  40                  45 Gly Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro 50                  55                  60 Tyr Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln 65                  70                  75                  80 Lys Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr 85                  90                  95 Phe Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu 100                 105                 110 Glu Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr 115                 120                 125 Lys Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser 130                 135                 140 Leu Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly 145                 150                 155                 160 Lys Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro 165                 170                 175 Asn His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His 180                 185                 190 Tyr Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys 195                 200                 205 Arg Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr 210                 215                 220 Pro Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu 225                 230                 235                 240 Tyr Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro 245                 250                 255 Asp Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe 260                 265                 270 Thr Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr 275                 280                 285 Phe Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly 290                 295                 300 Thr Leu Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu 305                 310                 315                 320 Tyr Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala 325                 330                 335 Ala Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile 340                 345                 350 Arg Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn 355                 360                 365 Phe Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys 370                 375                 380 Pro Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser 385                 390                 395                 400 Cys Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr 405                 410                 415 Asp Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe 420                 425                 430 Leu Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala 435                 440                 445 Ser Pro Ser Thr Leu Ser 450 ＜210＞    39 ＜211＞    452 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    39 Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp Asn 1               5                  10                  15 Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met 20                  25                  30 Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln 35                  40                  45 Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile 50                  55                  60 Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile 65                  70                  75                  80 Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr 85                  90                  95 Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp 100                 105                 110 Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn 115                 120                 125 Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr 130                 135                 140 Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp 145                 150                 155                 160 Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His 165                 170                 175 Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys 180                 185                 190 Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn 195                 200                 205 Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser 210                 215                 220 Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val 225                 230                 235                 240 Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala 245                 250                 255 Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp 260                 265                 270 Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly 275                 280                 285 Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu 290                 295                 300 Ser Ile Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met 305                 310                 315                 320 Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys 325                 330                 335 Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys 340                 345                 350 Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala 355                 360                 365 Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr 370                 375                 380 Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr 385                 390                 395                 400 Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala 405                 410                 415 Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys 420                 425                 430 Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro 435                 440                 445 Ser Thr Leu Ser 450 ＜210＞    40 ＜211＞    450 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    40 Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp Asn Ala Pro 1               5                  10                  15 Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp Met Ser Leu 20                  25                  30 Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly Gln Gly Val 35                  40                  45 Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr Ile Asp Ser 50                  55                  60 Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys Ile Ser Leu 65                  70                  75                  80 Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe Tyr Met Pro 85                  90                  95 Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu Trp Arg Pro 100                 105                 110 Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys Asn Arg Ser 115                 120                 125 Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu Thr Glu Ala 130                 135                 140 Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys Asp Phe Leu 145                 150                 155                 160 Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn His Leu Trp 165                 170                 175 Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr Lys Lys Pro 180                 185                 190 Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg Asn Asp Asp 195                 200                 205 Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro Ser Ile Tyr 210                 215                 220 Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr Val Arg Asn 225                 230                 235                 240 Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp Ala Lys Ser 245                 250                 255 Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr Asp Gln Val 260                 265                 270 Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe Gly Glu Thr 275                 280                 285 Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr Leu Ser Ile 290                 295                 300 Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr Met Asp Thr 305                 310                 315                 320 Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala Lys Met Cys 325                 330                 335 Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg Lys Asn Trp 340                 345                 350 Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe Ala Ile Gln 355                 360                 365 Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro Thr Leu Glu 370                 375                 380 Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys Tyr Ser Thr 385                 390                 395                 400 Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp Ala Val Asp 405                 410                 415 Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu Lys Pro Pro 420                 425                 430 Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser Pro Ser Thr 435                 440                 445 Leu Ser 450 ＜210＞    41 ＜211＞    428 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    41 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile 420                 425 ＜210＞    42 ＜211＞    429 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    42 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp 420                 425 ＜210＞    43 ＜211＞    430 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    43 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala 420                 425                 430 ＜210＞    44 ＜211＞    431 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    44 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Ser 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe 420                 425                 430 ＜210＞    45 ＜211＞    441 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    45 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr Glu Glu Pro 435                 440 ＜210＞    46 ＜211＞    443 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    46 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile 435                 440 ＜210＞    47 ＜211＞    445 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    47 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr 435                 440                 445 ＜210＞    48 ＜211＞    447 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    48 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala 435                 440                 445 ＜210＞    49 ＜211＞    449 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    49 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser 435                 440                 445 Pro ＜210＞    50 ＜211＞    451 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    合成序列 ＜400＞    50 Phe Arg Ala Pro Pro Val Ile Pro Asn Val Pro Phe Leu Trp Ala Trp 1               5                  10                  15 Asn Ala Pro Ser Glu Phe Cys Leu Gly Lys Phe Asp Glu Pro Leu Asp 20                  25                  30 Met Ser Leu Phe Ser Phe Ile Gly Ser Pro Arg Ile Asn Ala Thr Gly 35                  40                  45 Gln Gly Val Thr Ile Phe Tyr Val Asp Arg Leu Gly Tyr Tyr Pro Tyr 50                  55                  60 Ile Asp Ser Ile Thr Gly Val Thr Val Asn Gly Gly Ile Pro Gln Lys 65                  70                  75                  80 Ile Ser Leu Gln Asp His Leu Asp Lys Ala Lys Lys Asp Ile Thr Phe 85                  90                  95 Tyr Met Pro Val Asp Asn Leu Gly Met Ala Val Ile Asp Trp Glu Glu 100                 105                 110 Trp Arg Pro Thr Trp Ala Arg Asn Trp Lys Pro Lys Asp Val Tyr Lys 115                 120                 125 Asn Arg Ser Ile Glu Leu Val Gln Gln Gln Asn Val Gln Leu Ser Leu 130                 135                 140 Thr Glu Ala Thr Glu Lys Ala Lys Gln Glu Phe Glu Lys Ala Gly Lys 145                 150                 155                 160 Asp Phe Leu Val Glu Thr Ile Lys Leu Gly Lys Leu Leu Arg Pro Asn 165                 170                 175 His Leu Trp Gly Tyr Tyr Leu Phe Pro Asp Cys Tyr Asn His His Tyr 180                 185                 190 Lys Lys Pro Gly Tyr Asn Gly Ser Cys Phe Asn Val Glu Ile Lys Arg 195                 200                 205 Asn Asp Asp Leu Ser Trp Leu Trp Asn Glu Ser Thr Ala Leu Tyr Pro 210                 215                 220 Ser Ile Tyr Leu Asn Thr Gln Gln Ser Pro Val Ala Ala Thr Leu Tyr 225                 230                 235                 240 Val Arg Asn Arg Val Arg Glu Ala Ile Arg Val Ser Lys Ile Pro Asp 245                 250                 255 Ala Lys Ser Pro Leu Pro Val Phe Ala Tyr Thr Arg Ile Val Phe Thr 260                 265                 270 Asp Gln Val Leu Lys Phe Leu Ser Gln Asp Glu Leu Val Tyr Thr Phe 275                 280                 285 Gly Glu Thr Val Ala Leu Gly Ala Ser Gly Ile Val Ile Trp Gly Thr 290                 295                 300 Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile Lys Glu Tyr 305                 310                 315                 320 Met Asp Thr Thr Leu Asn Pro Tyr Ile Ile Asn Val Thr Leu Ala Ala 325                 330                 335 Lys Met Cys Ser Gln Val Leu Cys Gln Glu Gln Gly Val Cys Ile Arg 340                 345                 350 Lys Asn Trp Asn Ser Ser Asp Tyr Leu His Leu Asn Pro Asp Asn Phe 355                 360                 365 Ala Ile Gln Leu Glu Lys Gly Gly Lys Phe Thr Val Arg Gly Lys Pro 370                 375                 380 Thr Leu Glu Asp Leu Glu Gln Phe Ser Glu Lys Phe Tyr Cys Ser Cys 385                 390                 395                 400 Tyr Ser Thr Leu Ser Cys Lys Glu Lys Ala Asp Val Lys Asp Thr Asp 405                 410                 415 Ala Val Asp Val Cys Ile Ala Asp Gly Val Cys Ile Asp Ala Phe Leu 420                 425                 430 Lys Pro Pro Met Glu Thr Glu Glu Pro Gln Ile Phe Tyr Asn Ala Ser 435                 440                 445 Pro Ser Thr 450 ＜210＞    51 ＜211＞    435 ＜212＞    PRT ＜213＞    人工序列 ＜220＞ ＜223＞    Hyal1 ＜400＞    51 Met Ala Ala His Leu Leu Pro Ile Cys Ala Leu Phe Leu Thr Leu Leu 1               5                  10                  15 Asp Met Ala Gln Gly Phe Arg Gly Pro Leu Leu Pro Asn Arg Pro Phe 20                  25                  30 Thr Thr Val Trp Asn Ala Asn Thr Gln Trp Cys Leu Glu Arg His Gly 35                  40                  45 Val Asp Val Asp Val Ser Val Phe Asp Val Val Ala Asn Pro Gly Gln 50                  55                  60 Thr Phe Arg Gly Pro Asp Met Thr Ile Phe Tyr Ser Ser Gln Leu Gly 65                  70                  75                  80 Thr Tyr Pro Tyr Tyr Thr Pro Thr Gly Glu Pro Val Phe Gly Gly Leu 85                  90                  95 Pro Gln Asn Ala Ser Leu Ile Ala His Leu Ala Arg Thr Phe Gln Asp 100                 105                 110 Ile Leu Ala Ala Ile Pro Ala Pro Asp Phe Ser Gly Leu Ala Val Ile 115                 120                 125 Asp Trp Glu Ala Trp Arg Pro Arg Trp Ala Phe Asn Trp Asp Thr Lys 130                 135                 140 Asp Ile Tyr Arg Gln Arg Ser Arg Ala Leu Val Gln Ala Gln His Pro 145                 150                 155                 160 Asp Trp Pro Ala Pro Gln Val Glu Ala Val Ala Gln Asp Gln Phe Gln 165                 170                 175 Gly Ala Ala Arg Ala Trp Met Ala Gly Thr Leu Gln Leu Gly Arg Ala 180                 185                 190 Leu Arg Pro Arg Gly Leu Trp Gly Phe Tyr Gly Phe Pro Asp Cys Tyr 195                 200                 205 Asn Tyr Asp Phe Leu Ser Pro Asn Tyr Thr Gly Gln Cys Pro Ser Gly 210                 215                 220 Ile Arg Ala Gln Asn Asp Gln Leu Gly Trp Leu Trp Gly Gln Ser Arg 225                 230                 235                 240 Ala Leu Tyr Pro Ser Ile Tyr Met Pro Ala Val Leu Glu Gly Thr Gly 245                 250                 255 Lys Ser Gln Met Tyr Val Gln His Arg Val Ala Glu Ala Phe Arg Val 260                 265                 270 Ala Val Ala Ala Gly Asp Pro Asn Leu Pro Val Leu Pro Tyr Val Gln 275                 280                 285 Ile Phe Tyr Asp Thr Thr Asn His Phe Leu Pro Leu Asp Glu Leu Glu 290                 295                 300 His Ser Leu Gly Glu Ser Ala Ala Gln Gly Ala Ala Gly Val Val Leu 305                 310                 315                 320 Trp Val Ser Trp Glu Asn Thr Arg Thr Lys Glu Ser Cys Gln Ala Ile 325                 330                 335 Lys Glu Tyr Met Asp Thr Thr Leu Gly Pro Phe Ile Leu Asn Val Thr 340                 345                 350 Ser Gly Ala Leu Leu Cys Ser Gln Ala Leu Cys Ser Gly His Gly Arg 355                 360                 365 Cys Val Arg Arg Thr Ser His Pro Lys Ala Leu Leu Leu Leu Asn Pro 370                 375                 380 Ala Ser Phe Ser Ile Gln Leu Thr Pro Gly Gly Gly Pro Leu Ser Leu 385                 390                 395                 400 Arg Gly Ala Leu Ser Leu Glu Asp Gln Ala Gln Met Ala Val Glu Phe 405                 410                 415 Lys Cys Arg Cys Tyr Pro Gly Trp Gln Ala Pro Trp Cys Glu Arg Lys 420                 425                 430 Ser Met Trp 435

Claims (32)

1. 一種醫藥組合物，包含：(a)一藥物，為蛋白質藥物、抗體、小分子、適體、RNAi、反義RNA或細胞治療劑；以及(b)一PH20變異體，其中該PH20變異體在具有SEQ ID NO：1序列的一野生型PH20中包含M345T、K349E、L353A、L354I、N356E及I361T之胺基酸殘基取代。
2. 如請求項1所述之醫藥組合物，其中該PH20變異體更包含S343E之胺基酸殘基取代。
3. 如請求項1所述之醫藥組合物，其中該PH20變異體在選自由SEQ ID NO：1之該野生型PH20的一α螺旋域及對應該α螺旋域之一連接域的一區域組成之群組之一或多個區域中更包含一或多個胺基酸殘基取代。
4. 如請求項3所述之醫藥組合物，其中SEQ ID NO：1之該野生型PH20的該α螺旋域為一α螺旋8域(S347至C381)，該連接域為介於一α螺旋7與一α螺旋8之間的連接域(A333至R346)。
5. 如請求項4所述之醫藥組合物，其中該α螺旋域及對應該α螺旋域之該連接域為SEQ ID NO：1之該野生型PH20中之T341至N363、T341至I361、L342至I361、S343至I361、I344至I361、M345至I361或M345至N363。
6. 如請求項4所述之醫藥組合物，其中選自由SEQ ID NO：1之該野生型PH20的該α螺旋8域(S347至C381)及介於該α螺旋7與該α螺旋8之間的該連接域(A333至R346)組成之群組之一或多個區域係被人類玻尿酸酶-1(Hyal1)對應區域之胺基酸序列的一或多個胺基酸殘基所取代。
7. 如請求項1所述之醫藥組合物，其中該PH20變異體更包含在選自由T341、L342、S343、I344、S347、M348、L352、D355、E359及N363組成之群組之一或多個位置的一或多個胺基酸殘基取代。
8. 如請求項7所述之醫藥組合物，其中該PH20變異體更包含選自由T341S、L342W、S343E、I344N、S347T、M348K、L352Q、D355K、E359D及N363G組成之群組之一或多個胺基酸殘基取代。
9. 如請求項7所述之醫藥組合物，其中該PH20變異體包含選自S347T、M348K、L352Q、D355K及E359D之一胺基酸殘基取代。
10. 如請求項9所述之醫藥組合物，其中該PH20變異體更包含選自由T341S、L342W、S343E、I344N及N363G組成之群組之一或多個胺基酸殘基取代。
11. 如請求項10所述之醫藥組合物，其中該PH20變異體包含選自下列群組之任一胺基酸殘基取代： (a)T341S、L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T；(b)L342W、S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T；(c)M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T；(d)M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D、I361T及N363G；(e)I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T；以及(f)S343E、I344N、M345T、S347T、M348K、K349E、L352Q、L353A、L354I、D355K、N356E、E359D及I361T。
12. 如請求項1所述之醫藥組合物，其中該PH20變異體更包含在一羧基端及一胺基端之至少一者的一或多個胺基酸殘基的缺失。
13. 如請求項12所述之醫藥組合物，其中在該PH20變異體中，在選自由M1至P42組成之群組的一胺基酸殘基之前，藉由切除使在該胺基端的一或多個胺基酸殘基缺失。
14. 如請求項13所述之醫藥組合物，其中在該PH20變異體中，在胺基酸殘基L36、N37、F38、R39、A40、P41或P42之前，藉由切除使在該胺基端的一或多個胺基酸殘基缺失。
15. 如請求項12所述之醫藥組合物，其中在該PH20變異體中，在選自由V455至L509組成之群組的一胺基酸殘基之後，藉由切除使在該羧基端的一或多個胺基酸殘基缺失。
16. 如請求項15所述之醫藥組合物，其中在該PH20變異體中，在選自由V455至S490組成之群組之一胺基酸殘基之後，藉由切除使在該羧基端的一或多個胺基酸殘基缺失。
17. 如請求項16所述之醫藥組合物，其中在該PH20變異體中，在胺基酸殘基V455、C458、D461、C464、I465、D466、A467、F468、K470、P471、P472、M473、E474、T475、E476、P478、I480、Y482、A484、P486、T488、或S490之後，藉由切除使在該羧基端的一或多個胺基酸殘基缺失。
18. 如請求項1至17之任一項所述之醫藥組合物，其中該PH20變異體在該胺基端更包含一訊息肽，該訊息肽源自人類玻尿酸酶-1(Hyal1)、人類生長激素或人類血清白蛋白。
19. 如請求項1至11之任一項所述之醫藥組合物，其中該PH20變異體具有選自SEQ ID NO：5至SEQ ID NO：50之胺基酸序列的一胺基酸序列。
20. 如請求項19所述之醫藥組合物，其中該PH20變異體具有SEQ ID NO：44之序列。
21. 如請求項1所述之醫藥組合物，其中該藥物為一抗體、一可溶性受體或融合一可溶性受體與一Fc區域的一融合蛋白。
22. 如請求項21所述之醫藥組合物，其中該抗體結合到選自由下列抗原組成之群組之一或多個抗原：4-1BB、整合素、類澱粉蛋白β、血管生長素、血管生長素類似物3、B細胞活化因子(BAFF)、B7-H3、補體5、CCR4、CD3、CD4、CD6、CD11a、CD19、CD20、CD22、CD30、CD33、CD38、CD52、CD62、CD79b、CD80、CGRP、密連蛋白-18、補體因子D、CTLA4、DLL3、EGF受體、血友病因子、Fc受體、FGF23、葉酸受體、GD2、GM-CSF、HER2、HER3、干擾素受體、干擾素γ、IgE、IGF-1受體、介白素1、介白素2受體、介白素4受體、介白素5、介白素5受體、介白素6、介白素6受體、介白素7、介白素12/23、介白素13、介白素17A、介白素17受體A、介白素31受體、介白素36受體、LAG3、LFA3、NGF、PVSK9、PD-1、PD-L1、RANK-L、SLAMF7、組織因子、TNF、VEGF及vWF。
23. 如請求項21所述之醫藥組合物，其中該抗體為選自由下列抗體組成之群組之一或多者：utomilumab、natalizumab、etrolizumab、vedolizumab、bimagrumab、bapineuzumab、crenezumab、solanezumab、aducanumab、gantenerumab、AMG 780、MEDI 3617、nesvacumab、vanucizumab、evinacumab、tabalumab、lanalumab、belimumab、omburtamab、ravulizumab、eculizumab、mogamulizumab、otelixizumab、teplizumab、muromonab、tebentafusp、blinatumoma、REGN1979、ibalizumab、zanolimumab、itolizumab、efalizumab、inebilizumab、tafasitamab、loncastuximab tesirine、ocrelizumab、 ublituximab、obinutuzumab、ofatumumab、rituximab、tositumomab、ibritumomab tiuxetan、epratuzumab、inotuzumab ozogamicin、moxetumomab pasudotox、brentuximab vedotin、vadastuximab talirine、gemtuzumab ozogamicin、daratumumab、isatuximab、alemtuxumab、crizanlizumab、polatuzumab vedotin、galiximab、eptinezumab、fremanezumab、galcanezumab、erenumab、zolbetuximab、lampalizumab、tremelimumab、zalifrelimab、ipilimumab、rovalpituzumab tesirine、cetuximab、depatuxizumab、zalutumumab、necitumumab、panitumumab、emicizumab、nipocalimab、rozanolixizumab、burosumab、farletuzumab、mirvetuximab soravtansine、dinutuximab、naxitamab、otilimab、margetuximab、pertuzumab、trastuzumab、trastuzumab deruxtecan、trastuzumab emtansine、trastuzumab duocarmazine、patritumab、anifrolumab、emapalumab、ligelizumab、omalizumab、dalotuzumab、figitumumab、teprotumumab、gebokizumab、canakinumab、daclizumab、basiliximab、dupilumab、mepolizumab、reslizumab、benralizumab、clazakizumab、olokizumab、sirukumab、siltuximab、sarilumab、satralizumab、tocilizumab、REGN88、secukinumab、ustekinumab、briakinumab、lebrikizumab、tralokinumab、ixekizumab、bimekizumab、brodalumab、brazikumab、guselkumab、risankizumab、tildrakizumab、mirikizumab、nemolizumab、spesolimab、relatlimab、narsoplimab、fasinumab、tanezumab、alirocumab、evolocumab、 bococizumab、lambrolizumab、balstilimab、camrelizumab、cemiplimab、dostarlimab、prolgolimab、sintilimab、spartalizumab、tislelizumab、nivolumab、atezolizumab、avelumab、envafolimab、durvalumab、bintrafusp alpha、denosumab、elotuzumab、concizumab、marstacimab、infliximab、adalimumab、golimumab、certolizumab pegol、ozoralizumab、brolucizumab、ranibizumab、bevacizumab、faricimab、ramucirumab及caplacizumab。
24. 如請求項21所述之醫藥組合物，其中該可溶性受體或融合該可溶性受體與該Fc區域的該融合蛋白所包含之該可溶性受體係選自由TNF-α可溶性受體、VEGF可溶性受體、CTLA-4、介白素1可溶性受體及LFA3可溶性受體組成之群組。
25. 如請求項24所述之醫藥組合物，其中融合該可溶性受體與該Fc區域的該融合蛋白係選自由etanercept、aflibercept、abatacept、belatacept、rilonacept及alefacept組成之群組。
26. 如請求項1所述之醫藥組合物，更包含選自由一緩衝劑、一穩定劑及一界面活性劑組成之群組之一或多者。
27. 如請求項26所述之醫藥組合物，其中該緩衝液包含選自由蘋果酸鹽、甲酸鹽、檸檬酸鹽、乙酸鹽、丙酸鹽、吡啶、哌

    

    、二甲砷酸鹽、琥珀酸鹽、2-(N-

    

    啉基)乙磺酸(MES)、組胺酸、三羥甲基胺基甲烷(Tris)、雙-(2-羥基乙基)胺基三羥甲基胺基甲烷(bis-Tris)、磷酸鹽、乙醇胺、碳酸鹽、哌

    

    -N,N'-雙(2-乙 磺酸)(PIPES)、咪唑、1,3-雙(三(羥甲基)甲基胺基)丙烷(BIS-TRIS propane)、N,N-雙(2-羥乙基)-2-胺基乙磺酸(BES)、3-(N-

    

    啉基)丙磺酸(MOPS)、羥乙基哌

    

    乙磺酸(HEPES)、焦磷酸鹽及三乙醇胺組成之群組之一或多者；該穩定劑包含選自由一碳水化合物、一糖或其水合物、一糖醇或其水合物及一胺基酸組成之群組之一或多者；並且該界面活性劑包含選自由聚氧乙烯-山梨醇脂肪酸酯、聚乙烯-聚丙二醇、聚氧乙烯-硬脂酸酯、聚氧乙烯烷基醚、聚氧乙烯-聚氧丙烯共聚物及十二烷基磺酸鈉(SDS)組成之群組之一或多個非離子界面活性劑。
28. 如請求項27所述之醫藥組合物，其中該碳水化合物、該糖或該糖醇包含選自由海藻糖或其水合物、蔗糖、糖精、甘油、丁四醇、蘇糖醇、木糖醇、阿拉伯糖醇、核糖醇、甘露醇、山梨醇、半乳糖醇、海藻糖醇、艾杜糖醇、肌醇、庚七醇、異麥芽酮糖醇、麥芽糖醇、聚醣醇、環糊精、羥丙基環糊精及葡萄糖組成之群組之一或多者，並且該胺基酸包含選自由麩醯胺酸、麩胺酸、甘胺酸、離胺酸、離胺基離胺酸(lysilysine)、白胺酸、甲硫胺酸、纈胺酸、絲胺酸、硒甲硫胺酸、瓜胺酸、精胺酸、天冬醯胺酸、天冬胺酸、烏胺酸、異白胺酸、牛磺酸、茶胺酸、蘇胺酸、色胺酸、酪胺酸、苯丙胺酸、脯胺酸、吡咯離胺酸、組胺酸及丙胺酸組成之群組之一或多者。
29. 如請求項26所述之醫藥組合物，其中該醫藥組合物包含一組胺酸緩衝液、海藻糖及甲硫胺酸，該組胺酸緩衝液提供5.5±2.0之pH。
30. 如請求項26所述之醫藥組合物，其中該醫藥組合物包含一組胺酸緩衝液、海藻糖、甲硫胺酸及聚山梨醇酯，該組胺酸緩衝液提供5.5±2.0之pH。
31. 如請求項30所述之醫藥組合物，其中該醫藥組合物包含該組胺酸緩衝液、10-400mM α,α-海藻糖、1-50mM甲硫胺酸及0.0000001%(w/v)至0.5%(w/v)之聚山梨醇酯，該組胺酸緩衝液提供5.5±2.0之pH。
32. 一種用於皮下注射的注射製劑，該注射製劑包含如請求項1至31之任一項所述之醫藥組合物。

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