TW205005B

TW205005B -

Info

Publication number: TW205005B
Application number: TW080105045A
Authority: TW
Original assignee: Bristol Myers Squibb Co
Priority date: 1990-06-20
Filing date: 1991-06-28
Publication date: 1993-05-01
Also published as: WO1991019513A1; ZA914750B; IL98569A0

Description

205005 A 6 B6 經濟部中央標準局员工消f合作社印製五、發明説明（1 ) 1 .瀵諂本發明有關轉形生長因子一 /3 (TGF_;3)及TG F_/S拮抗劑對諝和血壓（BP)之用途。發明方法以實例説明，其中成熟之重組TGF — /?1 (ctTGF — /5) 用以快速降低成熟獮猴之血壓。但是，發現範畴不限於α TGF — /3 1之用途而包括有效調和血壓之TGF_ —冷群之所有成員的成熟及先質形式之用途，包括天然及重组之成熟 TGF-/31, TGF — /32, TGF — 召 3, TGF — /34等，以及TGF_/8雜種，類似物及潛在 TGF_;S複體。相同地，發明包括任何及所有具拮抗T GF — 活性之組成物，包括但不限於抗一 TGF — >3抗體及TGF-/8受體。 2 .發明背暑 2 · 1雜形牛唇因芊一厶 TGF — /3為最近所述調和細胞分化及增殖之多胜肽群之一種。此族中其他成員包括苗勒氏抑制物質（〇&166_ t al.，1986，Cell 45:685-698)，抑制素（Mason et al*» 1985，Nature 318:659-663)及由蜂蠅之 decapentapleg-ic基因複體之轉錄預測之蛋白質（Padgett et al.，1987, Nature 325：81-84)〇已確認四種TGF — /3並名為TGF — yS 1 , TGF-/32, TGF — >31. 2，及 TGF-;93。首先描述之一種，TGF- /9 1由兩個相同之具太蚯麿功闲+ KK«l^iiMCNSVP4規柊m〇x297公你) - 3 - (請先閲讀背面之注意事項再填寫本頁) 裝- 訂_ · 205005 經濟部屮央標準局員工消費合作社印製 A6 B6 五、發明説明（2 ) 1 3, 0 0 0分子量且由二硫鍵聯結之次單元組成（Asso-ian et al., 1983, J. Biol. Chem. 258:7155-7160; F-rolik et a 1. > 1983. Proc. Natl. Acad. Sc i· USA 80: 3676-3680； Frolik et al.. 1984t J. Biol. Chem· 260 :10995-11000)。其由各種組織純化包括胎盤（Frolik et al.，1983，Nature 325'81-84 ),血小板（Childs …et.^il. ，1982, Proc. Natl . Acad. Sci. USA 79：5312-5316； Assoian et al.， 1983» J. Biol. Chem. 258^7155-7160 )，腎臓（Roberts et al·，1983，Biochemistry 22:5692 -5698)，及脱礦骨（Seyedin et al.，1985，Proc. Natl· Acad.Sci·USA82:119-123)。已單離编碼人類（DeΓyn-ck et al·，1985，Nature 316:701-705),老鼠（Derync-k et al · , 1986, J . Biol. chem. 261:4377-4379)及人遠（Sharpies et al., 1987, DNA 6:239-244)之純種糸。此類純種条之DNA序列分析顯示TGF — /3 1是以大先質多肽合成，其羧端切除産生成熟之TGF — ；9單體。在所有上述來源之TGF — /3 1先質蛋白質中發現強烈之序列同質性。在1 0 %血清及表皮生長因子存在下，T G F — ；8 1 促進正常鼠腎纖維母細胞之非勾引性生長（Roberts et a-1., 1981, Proc. Natl. Acad. Sci. USA 78：5339-5343； Roberts et al.，1982. Nature 295 J 417-419 ϊ Twardz-ik et al., 1985, J. Cell. Biochem. 28:289-297);僅存有10%血清時，可誘致AKR—2B纖維母細胞之菌 (請先閲讀背面之注意事項再填窝本頁) 裝· 訂. 線< 太κ 疳 ΙΛ 用 Ψ 因 K Ψ4掛格（210x297公势） -4 - 經濟部屮央標準局員工消#合作社印製 205005 A6 _ B6 五、發明説明（3 ) 落形成（Tuckert et al.，1983，Cancer Res· 43:1518-1 586)。TGF—>S1顯然亦使幼鼠肌肉間質細胞分化産生軟骨特定巨細胞（Seyedin et al.，1986, J. Biol. Che-m. 261:5693-5695)〇與其對細胞增殖之影響不同的，純化自人類血小板之 T G F — /3 1在培養時顯然抑制特定細胞之生長（Jucky et al.， 1984， Science 226:705-707)。 T G F - /3 1 顯亦抑制多種人類痛細胞条之生長（Roberts et al., 1985, Prco. Natl. Acad. Sci. USA 82:119-123)。此種 T G F _/3 1之抑制/剌激效果依各種因素而定包括細胞種類及細胞生理狀態（評論參見Sporn et al., 1986, S-cience 233:532-534)〇 TGF — >32,如TGF — /91,為由兩個相同 13, 000道耳呑次單元雙硫鍵結组成之分子量 2 6, 0 0 0 的多肽（Chiefetz et al., 1987, Cel 1 48: 409-415； Ikeda et al·» 1987， Biochemistry 26：2406-2410),且單離自牛之脱礦骨（Seydin et al·, 1987, J. Biol. Chem· 262:1946-1949)，豬血小板 Chiefetz et a-1., 1987, 48:409-415),人類前列腺癌細胞条 ,P C — 3 (Ikeda eta 1· > 1987. biochemistry 26：2406 -2410),及人類神經真瘤細胞条（Wrann et al.，1987, ΕΜΒ0 6:1633-1636)。已單離编碼人類及人猿T G F — /3 之 cDNA 純種条（Madisen et al.，1988，DMA 7:1-8; Webb et al.，1988,DNA 7:493-497)。由兩個大的先質 (請先閲讀背面之注意事項再填寫本頁) 裝- 訂_ 線· 太《.後κ m Ψ因Η宅燋m(CNS)甲4姐格（210x297公修） -5 - 經濟部中央標準局員工消费合作社印製 205005 A6 _______B6 五、發明説明（4 ) 多肽之一切除成熟之TGF — /32,其mRNAs可經由分類接合産生（Webb et al., 1988, DNA 7:493-497)。 TGF-yS 1及TGF —点2在其成熟區中具7 1% 之胺基酸序列同一性，且其先質結構有41%之同一性。 TGF — /33,其胺基酸序列最近由cDNA純種糸衍出 ,顯示與TGF — /3 1及TGF — /32之成熟單匾具約 80%同質性之C—端1 1 2胺基酸序列（Dijke et al.， 1988，Proc. Natl . Acad . Sci . USA 85：4715-4719)〇 TGF- /3 1. 2為包括/3 1及；S2次單元（以二硫鍵聯結）之異二聚體形式（Chiefetz et al·, 1987，Cell 48: 409-415)。 3 ·發明镀論本發明有關使用TGF-/9多肽，TGF — >3拮抗劑，及/或其组合物調和血壓之方法。發明為了說明計可僅分為兩部分。首先，發明有關TGF — /3作為可迅速明顯降低血壓之抗高血壓劑之抗高血壓劑之用途。此部分之發明包括具高血壓活性之任何及所有T G F - /9多肽，包括成熟及先質形成之 TGF — /31、TGF-/S2、TGF — /33, 雜種TGF — /S4,潛在TGF —卢複體，TGF — /3類似物等。在發明特例中，藉實例更充分描述（部分6，下文中），人猿重組TGF — /3 1非經腸給藥以在獮猴中誘致動脈血壓之快速明顯，及持缠降低。相關實例中，太紙张β泞ΐΛΐίΐΨ团因玄熄姐格（210x297公舟） -β - (請先閲讀背面之注意事項再填寫本頁) 205005 經濟部屮央標準局貝工消费合作社印製五、發明説明（5 ) T G F — /3可在患有急性高血壓及伴有極度高血壓之緊急情況中用以將血壓迅速降至正常檫準。其次，發明有開TGF — /0拮抗劑之用途，其經由 T G F — >8及/或相關因素所致低血壓之抑制以提高血壓。任何拮抗TGF — /3活性之組成物可用於此，包括例如，抗一 TGF — /3抗鰻及TGF — /3受體。此外，.降低及 /或保持人體中循環TGF — /9濃度之方法造成類似之加壓效果。例如，抗一TGF — /3抗感應RNA分子可抑制生物活性TGF — /S之合成及釋出，由是防止過度訊息産生及形成之低血壓。 4.附圓説明人猿T G F —卢1 c一p巧A多核^酸序列及推演之胺基酸序列。PTGF_/?1_2之1600 B P 嵌入體繼代無性蘩殖成Ml 3mp 1 8及Ml 3mp 1 9 無性繁殖媒介（Y a n i s c h - P e r r ο η β t a 1 ♦，19 8 5，G e n e 3 3 : 103-119)且兩股皆使用二去氧錐終止法定序（Sanger et al·，1977，Proc · Natl* Acad· Sc i. USA 74·5463*5467 )。推演之人猿胺基酸序列直接示於cDNA序列上。人類TGF — /8 1核苷酸序列經校正並直接示於人猿 c DNA序列下，·打點表示序列中同質核苷酸殘基。人類及人猿蛋白質間之胺基酸差異示於上行。成熟之TGF-/3 1序列加框且訊息胜肽於上加線。 G~^ —- β 2 - 4 4 2 cDNA 之核苷太板移反泞ϋί闲Ψ团S3宅棵谁（CNS) Ψ4規核（210x297公婊） -7 (請先閲讀背面之注意事項再填窝本頁) 裝- 訂_ 線· 205005 A 6 B6 經濟部中央標準局負工消费合作社印製五、發明説明（6 ) |序列及之胺基_酸序到_1 P C 一 21之2597BP 嵌入體繼代無性繁殖成P EMBL (Dante et al .，1983, Nucleic Acids Res. 11:1645-1654)且二股上之序列皆使用二去氣鏈終止法定出（Sanger et al.，1977，Proc. Natl. Acad. Sci. USA 74:5463-5467)。顯示编碼序列且推演之胺基酸序列直示於上。成熟之TGF — /S 2.序列加框且訊息胜肽於上割線。潛在糖化部位示以星號。箭號顯示推論之訊息序列切除部位。人猿TGF- /32 — 4 14 之核苷酸序列同於人類TGF—32—442 cDNA 序列，除（a)去除核苷酸346至432 (括弧者）並代之一序列AAT,及（b)各種無轚核苷酸變化發生於結構中之其他部位（在變化核苷酸下直接以單一字母表示 )外。人猿TGF — /32 — 414先質之推演胺基酸序列同於人類TGF — /32 — 442先質胺基酸序列，除天門冬醛胺取代人類TGF — /82 — 442結構中之胺基酸殘基 1 16至 144外。人類TGF — ；92 — 414 c DNA之核苷酸序列經其下繼缠割線之區定序且發現與人類TGF — /32 — 442 cDNA序列具完美同一性，除去除核苷酸346至432並代之一序列AAT外。 1//32 先質 DAN 之核苷酸 — |, ----—— » '* " 序列及推構之胺基酸序列。顯示编碼序列並直接於上面顯 -'· ' ·-' 示推演之胺基酸序列。成熟之TGF — /S序列加框且先質訊息胜肽於上割線。糖化部位以星號表示。箭號表示推論之訊息序列切除部位。所示之TGF — /32成熟编碼序列太蚯张只团团玄坻rniCNS)平4姐格（210x297公:修） _ g - (請先閲讀背面之注意事項再填窝本頁) 裝- 訂- 線. 經濟部中央標準局負工消费合作社印製 205005 A6 _五、發明説明（7 ) 為人類者。人猿TGF- >32成熟编碼序列幾乎同於人類序列：僅有3個無磬鹽基變化且直接以單以字母示於變化之核苷酸之下。 5.發明詳沭本發明有蘭使用TGF — /9多胜肽，拮抗劑及/或其组合物在動物中調和血壓之方法。發明是基於非經腸給予成熟之rTGF — /8 1可迅速明明降低獮猴血壓之發現。因此，發明一方面有關TGF — /8 s作為抗高血壓/低血壓劑之用途。相對效應，即，提高及/或保持血壓，確實可藉可抑制TGF — 抗高血壓/低血壓效果之TGF — 泠拮抗劑完成。就此而論，發明包括可抑制TGF — >9 一誘致高血壓之抗一T G F _办抗體，T G F — /8受體及其他組成物之用途。 5 . 1 TGF-i8作為杭窩血颸劑之用涂發明一方面有闋TGF — 作為抗高血壓/低血壓劑之用途。申請人之原始數據顯示在人猿試驗體中生理可容受極限或附近劑量下r TGF — /3 1顯然可迅速明顯地降低血壓。就此而論，此劑量之TGF — /3非經腸給藥可為需要強勢治療之高血壓緊急情況所接受。較低劑量之 T G F — /3亦可減低血壓，此劑量亦適合中至嚴重高血壓之患者。此種患者中，可能期望較不強勢之治療，其中之相反副作用不能為人承受。太蚯张β泞ϋίϊίΙΦ因因玄熄m(CNS)V4捣格（210x297公修） —9 一 (請先閱讀背面之注意事項再填窝本頁) 装- 訂，線，經濟部中央標準局貝工消費合作社印製 205005 A 6 _B_6 五、發明説明（8 ) 舒張血壓大於130mmHg並患病諸高血壓性腦病 ,進行性腎衰竭，急性肺水腫，腦中風，視乳頭f腫或多重新生視網膜出血之患者通強以非經腸抗高血壓劑諸如，例如，硝基氫酸鹽及重氮氧化物強勢治療。此種急症性高血壓之治療通常針對於在3 0至6 0分鐘中將B P降至 1 0 0 m m H g ,因為迅速降低為患此緊急狀況之患者存活的閬鍵因素。 TGF — /3抗高血壓劑可單獨或與其他抗高血壓劑在適當藥學載體中經任何適當途徑給藥。高血壓緊急狀況中，非經腸給藥可最快降低BP,因此為此情況之建議給藥途徑。此外，TGF — /3可在體|铪藥前鍵結於載體或尋靶分子且/或摻入脂質體，撤囊，及控釋配方中。 5.1.1 T G F — θ 夾漉依本發明，成熟及/或先質形成之TGF — /3 1、 TGF-/82、 TGF — /83、 TGF-/91/;S2 等，可用以降低高血壓。所用之TGF — /8可得自各種來源，包括但不限於由適當來源單離TGF — /3,藉重組DNA 技術，或化學合成法産生TGF_yS等。 5 . 1 . 1 . 1 T G F - θ 1 天然TGF — yS 1可單離自各種來源。此種細胞行為之潛在調和劑藉各種正常及經轉形細胞在培養時合成（R〇-berts et al.,1981, Proc， Natl. Acad. Sci. USA 78:5 (請先閲讀背面之注意事項再蜞寫本頁) 裝. 訂- 線< 太R 疳镝用 Ψ 因 K Ψ4規格（210x297公；«·) -10 - 205005 A 6 B6 經濟部屮央標準局員工消费合作社印製五、發明説明（9 ) 339-5343)並已由各種來源包括胎盤（Frolik et al.，198 3, Proc. Natl. Acad. Sci. USA 80:3676-3680),腎（R-oberts et a 1. > 1983, Biochemistry 22:5692-5698), 尿（Twardzik et al., 1985，J. Cell. . Biochem. 28:289 -297)及血小板（Childs et al.，1982，Proc. Natl. Acad. Sci. USA 79:5312-5316)。此外，描述人類（Peryno-k et al·，1985，Nature 316:701-705)，鼠類（Derynck et al . , 1986，J . Biol. Chem. 261 ： 4377-4379),及猿類（Sharpies et al.， 1987， DNA 6:239-244) T G F — /3 1 純化。大量TGF — /S 1可藉重組DNA技術使用真核宿主細胞，其以含有表現調節單位所控制之TGF — /91编碼序列之重組DNA媒介轉移感染，得到。此法之例述於 1989年8月17日申請之共審理申請案 07/353, 728中，彼案在此完全併為參考。簡言之，编碼人猿TGF— >3 1先質之CDNA純種条得自由非洲綠猴細胞細，BSC—40所製之cDNA庫。示於圖1之成熟人猿TGF — /3 1推演胺基酸序列與成熟人類 TGF — /3 1者具100%同質性。構成表現媒介，其在 SV40表現單位控制下含有完整之人猿TGF_i8 1先質用之编碼序列。其用以轉移感染獮猴卵巢細胞（CHO 細胞）。形成之CHO轉移感染物主要産生並分泌高分子量複體，由彼藉例行酸化法釋出成熟生物活性之TGF—β 〇太泞诮 ffl 4»因 H 甲4组格（210x297公货） _ n _ (請先閲讀背面之注意事項再填窝本頁) 205005 A 6 B6 經濟部中央標準局貝工消费合作社印製五、發明説明（10) 5 . 1 · 1 . 2 Ύ G F - β Z 用於發明中之TGF — /92可得自各種來源。自牛脱礦骨單離之蛋白質確實與T GF —冷有閬（3^以丨114&-1·，1987，J· Biol. Chem. 262:1946-1949)。蛋白質亦單離自豬血小板（Cheifetz et al., 1987, Cell 48:409- « · · 415),人類前列腺癌細胞細糸PC — 3(Ikeda et al., 1987，Biochemistry 26:2406-2410),及人類神經真瘤細胞条（Wrann et al.，1987，EMBO 6:1633-1636)。此蛋白質之部分胺基酸序列顯示其同於T G F — /S且稱為T G F —β 2, 〇大量T G F _ /β可使用真核細胞，其以含由表現調節單位控制之TGF —；32编碼序列之重组DNA媒介轉移感染，藉重組DNA技術得到。此法之例述於1 989年 12月5日提申之共審理申請案07/446, 020, 彼案在此完全併為參考。簡言之，编碼人類TGF — /82 先質之c DNA純種条得自由經tamoxifen處理之人類前列腺癌細胞条PC — 3,之cDNA庫。此一純種条之 cDNA序列示於圔2並預期TGF_y32合成為442 胺基酸多肽先質，由彼藉蛋白水解切除衍生成熟之112 胺基酸TGF — /52次單元。此TGF — /92先質，名為 TGF->32 — 442,與 TGF — >31 有 41% 同質性。另一實例中，编碼人猿TGF — /32先質之cDNA純種条得自由非洲綠猴細胞条，BCS—40所製之太ΛϊΑιηΨ团斑格（210x297公贽） -12 - (請先閲讀背面之注意事項再填寫本頁) 205005 A 6 B6 經濟部中央標準局貝工消t合作社印製五、發明説明（11) cDNA庫。此純種条之一 cDNA序列預期 T G F — /3 2亦合成為4 1 4胺基酸多肽先質，由彼藉蛋白水解切除衍生成熟之1 12胺基酸TGF — /92次單元。此 TGF — /S2 先質，稱為 TGF — /S2 — 414,具有4 1 4胺基酸殘基之胺基酸序列且同於TGF-/32 — 442之胺基酸序列，除其含單一天門冬醛胺殘綦而非人類TGF — /52 — 442序列之1 16至135殘基數之 2 9胺基酸序列外。亦確認來自编碼人猿TGF—卢2—442先質之 BSC—40 cDNA庫之純種条以及來自编碼人類 TGF — /32 — 414 先質之人類 PC — 3 cDNA 庫之純種条。人類及人猿TGF — /32 — 442先質顯然在胺基酸標準為完美同質，如人類及人猿T G F — /3 2 — 4 1 4 般。 5.1.1.3雜種成孰及先質T G F —冷雜種成熟T G F 分子可使用重組DNA技術或合成法製備。此法之例亦述於1988年12月15曰提呈之共審理申請案284, 927,彼案在此完全併為參考 Ο 雜種先質T G F - /8分子可使用重組DNA技術或合成法製備，如1987年8月17日提呈之共審理申請案 07/353，728所述。簡言之，構成含有在相關聯結（SL,相同之轉釋密碼中）於TGF — /3訊息及先質序太站用ψ因囷玄摁迆（(：Ν5)Ψ4姐格（210x297公册） -13 - (請先閱讀背面之注意事項再填窝本頁) 205005 A 6 B6 經濟部中央標準局貝工消t合作社印製五、發明説明（12) 列之TGF — /92成熟编碼序列（見圖3)之表現媒介，並用以轉移感染獮猴卵巢細胞（CH ◦細胞）。形成之 CHO轉移感染物産生並分泌成熟，生物活性之TGF_β。 5 . 1 . 1.4 辞改皙之 TGF — iS ..... 本文所示不同TGF — /S分子所示之胺基酸序列變化 ,以及立體結構，鍵結胺基酸殘基之共價鍵種類，及/或胺基一或羧基一終端殘基之基圃添加的變化皆為本發明範畴。例如，發明所用之TGF - >3分子可包括交替序列諸如保留性變化*其導致無聲變化因而産生官能上相當之分子。因此，示於圖1一3之胺基酸序列可有多種變化諸如嵌入，切除及取代，或為保留性或為非保留性，而此變化可在其使用中提供特定優點。如本文所用，保留性取代包括所示序列中一或多锢胺基酸以另一値具相似極性及疏水性/親水性胺基酸取代，導置無轚置換及官能性相當之分子。此種保留性取代包括但不限於下群胺基酸中之取代：甘胺酸，丙胺酸；纈胺酸，異白胺酸，白胺酸；天門冬胺酸，穀胺酸；絲胺酸，蘇胺酸；賴胺酸，精胺酸；苯基丙胺酸，酪胺酸；及甲硫胺酸，正白胺酸。 5.1.1.5潜在TGF—占複餺 TGF — /9 1可單離自組織或組織培養細胞之非活化，生物潛型，其可藉Chaot rop i c agent，蛋白酶，或體内太切.张；？/?功用+因因它说维（〇^)平4相.格（210乂297公势） -14 - (請先閲讀背面之注意事項再場寫本頁) 裝· 訂- 線- 205005 A 6 B6 經濟部屮央標準局只工消費合作社印製五、發明説明（13) 活化。相同的，以人猿TGF_;S1先質编碼序列轉移感染之CHO细胞分泌高分子量澝在複體，其包括TGF— 先質之成熟及★前位〃區二者。TGF — /3先質之★前位〃區的伴隨出現亦可在單離自血小板之潛在TGF — /3 1複體中察得。雖然體内活化機構未知，但潛在複髖可提供TGF - /3 1生物活性之重要調節度。依發明方法，潛在TGF — /9複體可藉在天然體内活化機構之情況下釋出彼者而作為控制T G F — /8生物活性型所致之高血壓效應的工具。來自重組CHO細胞之潛在TGF — /3 1複體之確認，單離及定性更充分描述於1 989年8月17曰提呈之共審理申請案07/353, 728中，彼案在此完全併為參考。 5 . 2 TGF — /3桔抗割作為加壓皙的用涂申請人發現r TGF —々1可迅速明顯降低血壓顯示 BP調節及/或低血壓之産生中可能牽涉TGF —卢。就此而論，經由其阻礙TGF— /3之低血壓效應的能力， TGF —々之桔抗劑可用為可提高B P之加壓/減壓劑。有效拮抗TGF — >9之低血壓效應的任何組成物皆可用於此，包括但不限於抗一 TGF 8抗體及TGF — /3受體 Ο 例如，TGF — /3拮抗劑可用於喪失BP性之治療醫藥條件下，其中期望將BP提高至正常程度。此條件包括，例如，血量喪失所致之休克，心臓病危險，及急性腎衰 (請先閲讀背面之注意事項再填寫本頁) 裝· 訂_ 線. 太蚯张κ疳ϋί用Ψ因Η玄捃m(CNS)平4規格（210x297公贽） -15 - 205005 A 6 B6 經濟部中央標準局貝工消費合作社印製五、發明説明（14) 竭中之低血壓。TGF_ /3可單獨或與其他加壓質/減壓質諸如多坡明（dopamine),埃平弗因（epinephrine),胺非林（aminophylline)等組合且/或共同給藥。與適當藥學載體調製含有效劑量之TGF — >3拮抗劑之複合物可經任何適當途徑（包括但不限於注射，輸入及選擇性導入）給予患者低血壓或伴有低血壓症狀之患者以提高B P。_此外，TGF - /3拮抗劑可在鳢藥前聯結於載體或尋靶子且/或摻入脂質體，撤囊，及控釋配方。 5,2.1 抗一TGF — θ 杭賻可抑制T G F — /3之低血壓效應的抗體可用為加壓劑。各種技蓊界已知方法可用以産製針對TGF — /3抗原決定部位之多株抗體。就抗體産製而言，各種宿主動物可注射TGF — /3,或合成之TGF — /S胜肽以免疫，包括但不限於兔，小老鼠，大白鼠等。可用各種佐劑以增加免疫反應，依宿主種類，包括但不限於Freund’s (完全及不完全），無機膠諸如氫氣化鋁，表面活性物質諸如溶血卵磷脂，複合多元醇（pluronic polyol),多陰離子，配狀（p-eotide),油乳液，栓孔啉配（lyiBpet)血藍質，二硝基酚 ,及潛在可用之人類佐劑諸如BCG(Bacille Calmette-Guerin)及 Corynebacterium parvum。針對於TGF — /3抗原決定基之單株抗體可藉任何藉培養連缠細胞条産製單株抗體用之技術製備。此包括但不限於 Kohler and Milstein (1975，Nature 256，495-497 (請先閲讀背面之注意事項再填寫本頁) 太蚯袼泞ifi m Ψ03 63¾:熄miCNS)平4媒格（210x297公货）一 16 一 205005 經濟部中央標準局貝工消费合作社印製五、發明説明（15) )所述之雜種瘤技術，及最近之人類B-cell雜種瘤技術（K-osbor e t a 1 . , 1983，Immunology Today 4:72)與 EBV —雜種瘤技術（C o 1 e e t a 1 .，19 8 5，Μ ο η o c 1 ο n a 1 A n t i b o-dies and Cancer Therapy. Alan R· Liss， Inc.，pp. 77 -96)。含有遣傳性型分子之抗體碎片藉已知技術産生。例如，此類碎片包括但不限於F (ab /) 2碎片，其藉胃蛋白酶水解抗體分子産生，·可藉還原F (ab>) 2碎片之二硫鍵産生之Fab/碎片，及可藉以木瓜蛋白酶及還原劑處理抗體分子産生之兩F a b碎片。抗一 TGF —卢抗體之産生述於1 989年8月1 7 日申請之共審理申請案7/353, 728,及1989 年12月5日申請之共審理申請案07/446, 020 Ο 5.2.2 T G F - /3 夸體外源TGF — /3受體分子可為有用之加壓劑，因其可鍵循環之TGF_;S及/或外競之内源受體，其導發 TGF_y9之低血壓效應。TGF — /3可藉1 988年 11月11日所申請之共審理申請案269, 524中所述之方法製備，彼案在此完全併為參考。 6 .管施例：非經腸給藥之r TGF-θ 1對獨猴血限之 (請先閲讀背面之注意事項再填寫本頁) 裝- 線· 太饭珞K m巾因囷宅说迆（CNS)V4提格（210乂297公修）一 17 — 205005 經濟部中央標準局兵工消費合作社印製五、發明説明（16) 下述者為用以在毎日靜脈内輸予獮猴(Macaca f aso i -cularis)後評估r TGF — /3 1之藥物毒性效果的部分研究。在此所述之結果顯示r TGF - /3 1對血壓具有深厚之研究效果。 6.1方法步驟 .....6.1.1猫猴偵測一隻成熟雄及兩隻成熟雌獮猴來自毎日TGF— 召1處理之血壓變化。猴子飼以市售食品並隨意給水。經由在開始研究前至少5日外科植入之長期動脈導管量血壓。其中一隻雌猴（39 — 181)進行一般麻醉並在右骺靜脈植入長期靜脈導管。另一隻雌猴（2 9 — 8 2 5)及雄猴（B — 3 44)亦在髂動脈及靜脈中植入長期動脈及靜脈導管。導管經由靜脈注射用之澠管伸出以利試驗品或賦形劑對照物之給藥。 6.1.2試驗品配方試驗及/或對照品配方每日於給藥前完成。每 10mi試驗品給藥溶液（0. 0213mg/min藉混合 0. 66mi rTGF-/31 母液（0. 46mg /mi?在 5mM HC)2 中）與 9. 34m)2 在 PBS 溶液中之Ο.1%猴血清蛋白。每曰在調配後及給藥前記錄給藥溶液之pH。基於最近未插管之體重並約略最近之 0 . 1 m i?計算劑量。太认张Κ 因因iliSmfCNSVP/l掛格（210X297公炻） _ jg _ (請先閱讀背面之注意事項再填寫本頁) 裝- 線· 經濟部中央標準局貝工消f合作社印製 205005 A6 __B_6_ 五、發明説明（17) 重組成熟之TGF — /3 1由轉移感染以完全人猿 TGF- /3 1先質编碼序列之獮猴卵巢細胞的培養物上清液箪離純化，如1989年8月17日申請之共審理申請案07/353, 728所述，其在此完全併為參考。 6.1.3廉理猴39 — 1 8 1接受毎日8m)2/kg體積在PBS 中之1%猴蛋白（賦形劑對照組）連缠5日。猴69— 168以劑量0.17mg/kg及濃度 0. 0213mg/m 又連缅處理 5 日。rTGF — /31 以0. 51mg/kg劑量及0. 0639mg/m又濃度給予猴29—825, 39—181及B344。猴 29_829處理連缠3日，猴39-181連續3日，猴B — 344 1日。用以計算試驗品或對照组劑量之體重為未埋入管裝置所得之最近體重。 r TGF —々1或賦形劑對照組經長期靜脈管以 1 . 6 0 m $給藥溶液/分鐘經輸入泵（Harvard Appara-tuS)靜脈内注射，並經試驗備之檫準操作法校正。給予試驗品及對照物前，經0. 9%滅菌後之鹽水定期清洗導管以保持導管之明顯度。記錄r TGF — /3 1及對照組給藥之體積，時間，及日期。 6.1.4 rfn懕測鼉血壓測量經長期動脈管由猴29—825, 39— 太呔稂R泞边用Ψ因因犮德迆- 19 - (請先閲讀背面之注意事項再填寫本頁) 裝. 線< 205005 A 6 B 6 五、發明説明（18) 1 8 1及B344記錄。血壓測量在給予試驗品前及完成後至少持缠1分鐘。血壓測量亦記為修正者或依研究指導者之指示記錄，若此測量為臨床相關性。 6. 2臨庆觀察毎日觀察猿猴之臨床異常，食物及水攝取，辑溫，呼吸率，血壓，及其他變數29日。此外，由各猿猴收集血液試樣作為血液學，血清化學及免疫分析用之試樣。 6.2.1 1^丁0卩一/31對8卩之效果接受t TGF — /3 1注射之三猴中之二者顯示適當，明顯及持續之動脈血壓降低。第三隻實驗猴亦有BP損失 ,但此結果就給藥前此動物所具之極低血壓而言稍難判斷。在對照猴中亦觀察明顯之BP波動。此三實驗猴之個別 B P觀察示於表I。 • · · ..........·(.........裝.....玎· · · ·線·· (請先閲讀背面之注意事項再填寫本頁) 經濟部屮央標準局貝工消f合作社印製太冷iA Ψ因03¾:捣逛（CNS)甲4規格（210x297公焙） -20 - 205005 A6 B6 五、發明説明（19) 表 u.l以TGF—jg 1處理：>雄及雌獮猴之楠別ifn壓一袢g 動物编號性別劑量 (mg/kg) 研究曰餐時動脈血壓 (mm Hg) 39-181205005 A 6 B6 Printed by the Employees ’Cooperative of the Central Bureau of Standards of the Ministry of Economy V. Description of the invention (1) 1. This invention relates to the transforming growth factor 1/3 (TGF_; 3) and TG F_ / S antagonist antagonists. And blood pressure (BP) use. The method of the invention is illustrated by an example, in which the mature recombinant TGF — /? 1 (ctTGF — / 5) is used to rapidly reduce the blood pressure of the mature marmoset monkey. However, the scope of the discovery is not limited to the use of α TGF — / 3 1 but includes the use of the mature and progenitor forms of all members of the cold group TGF_ effective to regulate blood pressure, including natural and recombinant mature TGF- / 31, TGF — / 32, TGF — Zhao 3, TGF — / 34, etc., as well as TGF_ / 8 hybrids, analogs and potential TGF_; S complexes. Similarly, the invention includes any and all compositions with TGF-antagonizing activity, including but not limited to anti-TGF-> 3 antibodies and TGF- / 8 receptors. 2. The invention of backstroke 2 · 1 Miscellaneous bull's lips due to Qianyi Ji TGF — / 3 is one of the multi-peptide group that recently regulated cell differentiation and proliferation. Other members of this family include Muller's inhibitory substances (〇 & 166_t al., 1986, Cell 45: 685-698), inhibin (Mason et al * »1985, Nature 318: 659-663) and Youbei A protein predicted by transcription of the decapentapleg-ic gene complex of flies (Padgett et al., 1987, Nature 325: 81-84). Four TGF — / 3 have been identified and named TGF — yS 1, TGF- / 32, TGF — > 31. 2, and TGF-; 93. The first one is described. TGF- / 9 1 consists of two identical devices with the same function: KK «l ^ iiMCNSVP4 regulation 柊 m〇x297 you)-3-(please read the precautions on the back before filling this page ) Installation-Order _ · 205005 A6 B6 printed by the Employee Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (2) 1 3, 0 0 0 Molecular weight and composed of subunits linked by disulfide bonds (Asso-ian et al ., 1983, J. Biol. Chem. 258: 7155-7160; F-rolik et a 1. > 1983. Proc. Natl. Acad. Sc i · USA 80: 3676-3680; Frolik et al .. 1984t J . Biol. Chem. 260: 10995-11000). It is purified from various tissues including placenta (Frolik et al., 1983, Nature 325'81-84), platelets (Childs ... et. ^ Il., 1982, Proc. Natl. Acad. Sci. USA 79: 5312-5316; Assoian et al., 1983 »J. Biol. Chem. 258 ^ 7155-7160), nephrolithiasis (Roberts et al., 1983, Biochemistry 22: 5692-5698), and demineralized bone (Seyedin et al., 1985, Proc . Natl · Acad.Sci · USA82: 119-123). Coded humans (DeΓyn-ck et al., 1985, Nature 316: 701-705), mice (Derync-k et al., 1986, J. Biol. Chem. 261: 4377-4379) and humans (Sharpies et al., 1987, DNA 6: 239-244). DNA sequence analysis of such pure-breeding bars shows that TGF — / 3 1 is synthesized from a large precursor peptide, and its carboxyl end is excised to produce mature TGF —; 9 monomers. A strong sequence homogeneity was found in the TGF — / 3 1 precursor protein of all the above sources. In the presence of 10% serum and epidermal growth factor, TGF-; 8 1 promotes the non-sedulous growth of normal rat kidney fibroblasts (Roberts et a-1., 1981, Proc. Natl. Acad. Sci. USA 78: 5339-5343; Roberts et al., 1982. Nature 295 J 417-419 ϊ Twardz-ik et al., 1985, J. Cell. Biochem. 28: 289-297); when there is only 10% serum, it can be induced AKR-2B fibroblast cell bacteria (please read the precautions on the back before filling the nest page) Binding · Order. Thread & Too κ 疳 ΙΛ Use Ψ due to K Ψ4 hanging grid (210x297 public potential) -4-Ministry of Economic Affairs Printed by the staff of the Central Standards Bureau # 205 Cooperative Society 205005 A6 _ B6 V. Description of the invention (3) Formation (Tuckert et al., 1983, Cancer Res. 43: 1518-1 586). TGF— > S1 obviously also differentiates muscle stromal cells of young rats to produce cartilage-specific giant cells (Seyedin et al., 1986, J. Biol. Che-m. 261: 5693-5695). It has different effects on cell proliferation In fact, TGF — / 3 1 purified from human platelets obviously inhibits the growth of specific cells during culture (Jucky et al., 1984, Science 226: 705-707). TG F-/ 3 1 also inhibits the growth of various human pain cell strips (Roberts et al., 1985, Prco. Natl. Acad. Sci. USA 82: 119-123). The inhibitory / stimulating effect of such TGF_ / 3 1 depends on various factors including cell type and cell physiological state (for review see Sporn et al., 1986, S-cience 233: 532-534). TGF — > 32 , Such as TGF — / 91, is a polypeptide with a molecular weight of 2, 6, 0 0 0 composed of two identical 13,000 Daud subunit disulfide bonds (Chiefetz et al., 1987, Cel 1 48: 409-415 ; Ikeda et al · »1987, Biochemistry 26: 2406-2410), and demineralized bone isolated from cattle (Seydin et al ·, 1987, J. Biol. Chem · 262: 1946-1949), Piget Platelet Chiefetz et a-1., 1987, 48: 409-415), human prostate cancer cell strips, PC — 3 (Ikeda eta 1 · > 1987. biochemistry 26: 2406 -2410), and human neuromyoma cell strips (Wrann et al., 1987, EMB0 6: 1633-1636). The pure cDNA strips encoding human and ape TG F — / 3 have been isolated (Madisen et al., 1988, DMA 7: 1-8; Webb et al., 1988, DNA 7: 493-497). Consists of two large predecessors (please read the precautions on the back and then fill out this page) to install-order _ line · too ". After κ m Ψ due to Η 宅燋 m (CNS) A 4 sister grid (210x297 public repair) -5-Printed 205005 A6 _______B6 by the Staff Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economy V. Description of the invention (4) One of the peptides is excised from mature TGF — / 32, and its mRNAs can be generated by classification (Webb et al., 1988, DNA 7: 493-497). TGF-yS 1 and TGF-point 2 have 71% amino acid sequence identity in their mature regions, and their precursor structures have 41% identity. TGF — / 33, the amino acid sequence of which was recently derived from cDNA pure-breeding Shitou, shows that the mature single plaque with TGF — / 3 1 and TGF — / 32 has about 80% homology to the C-terminal 1 1 2 amino group Acid sequence (Dijke et al., 1988, Proc. Natl. Acad. Sci. USA 85: 4715-4719). TGF- / 3 1.2 includes / 3 1 and S2 subunits (linked by disulfide bonds) Heterodimer form (Chiefetz et al., 1987, Cell 48: 409-415). 3. Plating Theory of the Invention The present invention relates to a method for regulating blood pressure using TGF- / 9 polypeptide, TGF — > 3 antagonist, and / or a combination thereof. The invention can only be divided into two parts for the purpose of illustration. First, the invention relates to the use of TGF / 3 as an antihypertensive agent that can rapidly and significantly lower blood pressure. The inventions in this part include any and all TGF- / 9 polypeptides with hypertensive activity, including mature and precursor-forming TGF — / 31, TGF- / S2, TGF — / 33, hybrid TGF — / S4, potential TGF — Lu Fu body, TGF — / 3 analogs, etc. In the special case of the invention, more fully described by way of example (Part 6, below), the parental administration of recombinant TGF — / 3 1 in apes induces rapid and obvious arterial blood pressure and reduced entanglement in marmosets. In the related example, the Tai paper β 択 ΐΛΙίΐΨ 團 because of mysterious extinction (210x297 Gongzhou) -β-(please read the precautions on the back and then fill out this page) 205005 Printed by Beigong Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (5) TGF / 3 can be used to rapidly lower blood pressure to normal spurs in emergency situations with acute hypertension and extreme hypertension. Secondly, the invention has the use of prescribing TGF — / 0 antagonists, which can increase blood pressure by suppressing hypotension caused by T G F —> 8 and / or related factors. Any composition that antagonizes TGF-3 activity can be used here, including, for example, anti-TGF-3 anti-eel and TGF-3 receptors. In addition, the method of reducing and / or maintaining the circulating TGF— / 9 concentration in the human body results in a similar boosting effect. For example, anti-TGF — / 3 anti-sensing RNA molecules can inhibit the synthesis and release of biologically active TGF — / S, thereby preventing the generation and formation of excessive blood pressure from hypotension. 4. Attached to the circle description Apes T G F —Lu 1 c-p Qiao A polynucleotide sequence and deduced amino acid sequence. The 1600 BP inserts of PTGF _ /? 1_2 were cloned into Ml 3mp 1 8 and Ml 3mp 1 9 asexual propagation medium (Y anisch-P err ο η β ta 1 ♦, 19 8 5, G ene 3 3 : 103-119) and the use of two deoxygenated cones to terminate the legal order (Sanger et al., 1977, Proc. Natl * Acad. Sc i. USA 74. 5463 * 5467). The deduced amino acid sequence of simian is shown directly on the cDNA sequence. The human TGF — / 8 1 nucleotide sequence is corrected and displayed directly under the simian c DNA sequence, and dotted dots represent homologous nucleotide residues in the sequence. The differences in amino acids between human and ape proteins are shown on the top. The mature TGF- / 3 1 sequence is framed and the message peptide is added on the line. G ~ ^ —- β 2-4 4 2 The nucleoside of the cDNA is transferred from the nucleoside plate to the S3 house who is in the S3 house (CNS) Ψ4 regulation core (210x297 public bitch) -7 (please read the precautions on the back first Fill this page) Binding-Order _ Line · 205005 A 6 B6 Printed by the Ministry of Economic Affairs Central Standards Bureau Negative Work Consumer Cooperative V. Invention Instructions (6) | Sequence and Amino Groups_ Acid Order to _1 PC One 21 of 2597BP The embedding clones were cloned into P EMBL (Dante et al., 1983, Nucleic Acids Res. 11: 1645-1654) and the sequences on the two strands were legally released using two degassing chain terminations (Sanger et al., 1977, Proc. Natl. Acad. Sci. USA 74: 5463-5467). The coding sequence is shown and the deduced amino acid sequence is shown directly above. Mature TGF — / S 2. The sequence is framed and the message peptide is cut on the upper line. The potential saccharification sites are shown with an asterisk. The arrow shows the excision site of the inferred message sequence. The nucleotide sequence of TGF- / 32 — 4 14 is the same as the cDNA sequence of human TGF—32—442, except (a) nucleotides 346 to 432 (brackets) are removed and a sequence AAT is substituted, and (b) Various non-successful nucleotide changes occur in other parts of the structure (directly represented by a single letter under the changed nucleotide). The deduced amino acid sequence of the precursor TGF — / 32 — 414 is the same as that of the human TGF — / 32 — 442, except that the asparagine replaces the amino acid in the structure of human TGF — / 82 — 442 Residues 1 16 to 144. The nucleotide sequence of human TGF —; 92 — 414 c DNA is sequenced by the area following the tangent line and found to have perfect identity with the human TGF — / 32 — 442 cDNA sequence, except that nucleotides 346 to 432 are removed Generation one sequence outside AAT. 1 // 32 The nucleotide of precursor DAN — |, ----—— »'* " sequence and deduced amino acid sequence. Display the coding sequence and directly display-'·' ·-'above to indicate the deduced amino acid sequence. The mature TGF — / S sequence is framed and the precursor message is on the upper secant. The saccharification site is indicated by an asterisk. The arrow indicates the location of the deduced message sequence. The TGF — / 32 mature coding sequence shown is too earthworm Zhang Zhituan Xuandi rniCNS) Ping 4 sister grid (210x297 g: repair) _ g-(please read the precautions on the back and fill in this page) -Line. Printed 205005 A6 by the Consumer Labor Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs. V. Description of Invention (7) For human beings. The ape TGF- > 32 mature coding sequence is almost the same as the human sequence: there are only three non-salt base changes and are shown directly under the changed nucleotides in single letters. 5. Details of the invention The present invention has a method for regulating blood pressure in animals using TGF- / 9 polypeptide, antagonists and / or combinations thereof. The invention is based on the discovery that parenteral administration of mature rTGF — / 8 1 can quickly and clearly reduce blood pressure in marmosets. Therefore, one aspect of the invention relates to the use of TGF / 8 s as an antihypertensive / hypertensive agent. The relative effect, that is, raising and / or maintaining blood pressure, can indeed be accomplished with TGF-ling antagonists that can inhibit TGF—antihypertensive / hypertensive effects. In this connection, the invention includes the use of anti-T G F _ antibody, T G F / 8 receptor and other components that can inhibit TGF —> 9 — induced hypertension. 5.1. Application of TGF-i8 as an anti-hypertensive / hypotensive agent for the application of Hangwo blood scavenger. The original data of the applicant shows that r TGF — / 3 1 at the physiologically tolerable limit or near dose in the anthropoid test subjects can obviously and rapidly lower blood pressure. In this regard, parenteral administration of this dose of TGF / 3 can be accepted for emergency treatment of hypertension requiring strong treatment. Lower doses of T G F — / 3 can also lower blood pressure, and this dose is also suitable for patients with moderate to severe hypertension. In such patients, less powerful treatment may be expected, and the opposite side effects cannot be tolerated. Taizhang Zhang β 泞 ϋίϊίΙΦ due to mysterious methane (CNS) V4 stamping (210x297 public repair) — 9 1 (please read the precautions on the back before filling the nest page) Binding-Order, Line, Central Bureau of Standards, Ministry of Economic Affairs Printed by Beigong Consumer Cooperative Society 205005 A 6 _B_6 V. Description of the invention (8) Diastolic blood pressure greater than 130 mmHg and suffering from various hypertensive encephalopathy, progressive renal failure, acute pulmonary edema, cerebral stroke, optic papilloma or multiple newborn retina Hemorrhagic patients are usually treated with parenteral antihypertensive agents such as, for example, nitrohydrogenate and diazo oxide. This type of treatment for acute hypertension is usually aimed at reducing B P to 100 mm Hg in 30 to 60 minutes, because the rapid reduction of the factor of survival for patients suffering from this emergency. TGF— / 3 antihypertensive agents can be administered by any suitable route alone or in combination with other antihypertensive agents in a suitable pharmaceutical carrier. In emergency situations of hypertension, parenteral administration can reduce BP the fastest, so the recommended route of administration for this situation. In addition, TGF-3 can be bound to carriers or targeting molecules before the body | hafnium drugs and / or incorporated into liposomes, withdrawn capsules, and controlled release formulations. 5.1.1 TGF — θ In accordance with the present invention, mature and / or precursor formed TGF — / 3 1, TGF- / 82, TGF — / 83, TGF- / 91 /; S2, etc., can be used to reduce hypertension . The TGF — / 8 used can be obtained from various sources, including but not limited to isolation of TGF — / 3 from an appropriate source, TGF_yS produced by recombinant DNA technology, or chemical synthesis. 5.1.1.1 T G F-θ 1 natural TGF — yS 1 can be isolated from various sources. Potential mediators of this cell behavior are synthesized by various normal and transformed cells during culture (R〇-berts et al., 1981, Proc, Natl. Acad. Sci. USA 78: 5 (please read the notes on the back Matters will be written on this page). Packing. Order-Line < Too R for dysprosium due to K Ψ4 specifications (210x297 g; «·) -10-205005 A 6 B6 Printed by the Employee Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. , Description of the invention (9) 339-5343) and has been from various sources including placenta (Frolik et al., 198 3, Proc. Natl. Acad. Sci. USA 80: 3676-3680), kidney (R-oberts et a 1 > 1983, Biochemistry 22: 5692-5698), urine (Twardzik et al., 1985, J. Cell.. Biochem. 28: 289 -297) and platelets (Childs et al., 1982, Proc. Natl. Acad . Sci. USA 79: 5312-5316). In addition, describe humans (Peryno-k et al., 1985, Nature 316: 701-705), rodents (Derynck et al., 1986, J. Biol. Chem. 261: 4377-4379), and apes (Sharpies et al., 1987, DNA 6: 239-244) TGF — / 3 1 purification. A large number of TGF — / S 1 can be obtained by recombinant DNA technology using eukaryotic host cells, which are transferred and infected with recombinant DNA vectors containing TGF — / 91 coding sequences controlled by expression regulatory units. An example of this law is described in the co-pending application 07/353, 728 filed on August 17, 1989, which is fully incorporated herein by reference. In short, the pure DNA strips of CDNA encoding the apes TGF-> 3 1 precursor are free from the cDNA library made by BSC-40 of African green monkey cells. The deduced amino acid sequence of mature ape TGF — / 3 1 shown in Figure 1 is 100% homogenous with that of mature human TGF — / 3 1. It constitutes the expression medium, which contains the complete coding sequence for the apes TGF_i8 1 precursor under the control of the SV40 expression unit. It is used to transfer infection to marmoset ovarian cells (CHO cells). The formed CHO transfer infection mainly produces and secretes high-molecular-weight complexes, and the mature biologically active TGF-β is released by him through routine acidification method. 汞泞诮 ffl 4 n _ (Please read the precautions on the back before filling the nest page) 205005 A 6 B6 Printed by the Beigong Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs V. Invention Instructions (10) 5.1.1.2 Ύ GF-β Z TGF — / 92 used in the invention can be obtained from various sources. The protein isolated from bovine demineralized bone is indeed different from T GF-cold and cold (3 ^ Yi Shu 114 & -1 ·, 1987, J · Biol. Chem. 262: 1946-1949). The protein is also isolated from porcine platelets (Cheifetz et al., 1987, Cell 48: 409- «· 415), human prostate cancer cell line PC-3 (Ikeda et al., 1987, Biochemistry 26: 2406-2410) , And human neuromyoma cell strips (Wrann et al., 1987, EMBO 6: 1633-1636). Part of the amino acid sequence of this protein shows that it is the same as TGF — / S and is called TGF — β 2, 〇 A large number of TGF _ / β can be used in eukaryotic cells, which contains TGF controlled by performance regulating units —; 32 The infection of the recombinant DNA vector of the code sequence is obtained by recombinant DNA technology. An example of this law is described in the application for co-examination on December 5, 1 989, 07/446, 020. This case is fully incorporated herein by reference. In short, the pure DNA strip of c DNA encoding human TGF — / 82 precursor is obtained from the cDNA library of PC-3, a human prostate cancer cell strip treated with tamoxifen. The cDNA sequence of this pure seed bar is shown in 圔 2 and TGF_y32 is expected to be synthesized as 442 amino acid polypeptide precursor, which is proteolytically cleaved to derive the mature 112 amino acid TGF — / 52 subunit. This TGF — / 92 precursor, named TGF-> 32 — 442, has 41% homogeneity with TGF —> 31. In another example, the pure cDNA encoding the ancestor TGF — / 32 precursor is obtained from African green monkey cell strips, and the ΛϊΑιηΨ cluster patch (210x297 Gongzhi) made by BCS-40 -12-(please read first (Notes on the back and then fill out this page) 205005 A 6 B6 Printed by the Beigongxiaot Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (11) cDNA library. The cDNA sequence of one of the purebreds is expected to be TG F — / 3 2 also synthesized into 4 1 4 amino acid peptide precursors, which are hydrolyzed and cleaved from the protein to derive the mature 1 12 amino acid TGF — / 92 subunit. This TGF — / S2 precursor, called TGF — / S2 — 414, has an amino acid sequence of 414 amino acid residues and is the same as the amino acid sequence of TGF- / 32 — 442, except that it contains a single Asparagine residues are not human TGF — / 52 — 442 sequence 1 16 to 135 residue number 2 9 amino acid sequence. Also confirmed are the purebred strips from the BSC-40 cDNA library encoding the ape TGF—Lu 2-442 precursor and the purebred strips from the human PC-3 cDNA library encoding the human TGF— / 32—414 precursor. The human and ape TGF — / 32 — 442 precursor is obviously perfectly homogenous at the amino acid standard, as is the case for human and ape T G F — / 3 2 — 4 1 4. 5.1.1.3 Hybrid formation and precursor TG F-cold Hybrid mature TG F molecules can be prepared using recombinant DNA technology or synthetic methods. Examples of this method are also described in the co-trial application filed on December 15, 1988, 284, 927, the case is complete and for reference Ο Hybrid precursor TGF-/ 8 molecules can be prepared using recombinant DNA technology or synthesis , As described in the co-pending application 07 / 353,728 submitted on August 17, 1987. In short, the composition is contained in the related link (SL, the same transfer code) in the TGF — / 3 message and the pre-primary order is used by the station ψ due to mystery ((: Ν5) Ψ4 sister grid (210x297 book ) -13-(Please read the precautions on the back before filling the nest page) 205005 A 6 B6 Printed by Beigongxiaot Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs V. TGF — / 92 mature code listed in (12) Sequence (see Figure 3) expression medium, and used to transfer infection of marmoset ovarian cells (CH ◦ cells). The formed CHO transfer infection produces and secretes mature, biologically active TGF_β. 5.1.1.4 Resignation of TGF — IS ..... The amino acid sequence changes shown in the different TGF — / S molecules shown in this article, as well as the three-dimensional structure, the type of covalent bond bonding the amino acid residue, and / or the amino group or carboxyl group Variations of the addition of a terminal residue to the base are within the scope of the invention. For example, the TGF-> 3 molecules used in the invention may include alternating sequences such as retention changes * which result in silent changes and thus produce functionally equivalent molecules. Therefore , The amino acid sequence shown in Figures 1 to 3 can have many changes such as embedding Excision and substitution, either reserved or non-reserved, and this change can provide specific advantages in its use. As used herein, reserved substitutions include one or more amino acids in the sequence shown as another Similar polar and hydrophobic / hydrophilic amino acid substitutions, leading to non-replaceable substitutions and functionally equivalent molecules. Such reserved substitutions include but are not limited to substitutions in the following group of amino acids: Glycine, Alanine; Valine, isoleucine, leucine; aspartic acid, glutamic acid; serine, threonine; lysine, arginine; phenylalanine, tyrosine; and methylthio Amino acid, n-leucine. 5.1.1.5 Potential TGF—Zhan Fujun TGF— / 9 1 can be isolated from tissues or tissue culture cells without activation, biologically latent, which can be borrowed from Chaot rop ic agent, protease, or The body is too cut. Zhang;? /? Function + because it says to maintain (〇 ^) flat 4 phases. Grid (210 297 public power) -14-(please read the precautions on the back before writing this page) Binding · Ordering-Line-205005 A 6 B6 Printed by the Workers and Consumers Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs V. Invention Instructions (13) Activation. Phase The CHO cells transfected with the coding sequence of the apes TGF_; S1 secrete high molecular weight secreted in the complex, which includes both the mature of the TGF-precursor and the anterior 〃 region. TGF — / 3 of the precursor The concomitant appearance of the anterior 〃 region can also be detected in latent platelet-derived potential TGF — / 3 1 complex. Although the activation mechanism in vivo is unknown, the potential hip complex can provide important regulation of TGF-/ 3 1 biological activity According to the method of the invention, the potential TGF — / 9 complex can be used as a tool to control the hypertensive effect caused by the TGF — / 8 bioactive type by releasing the other in the context of the natural in vivo activation mechanism. The identification, isolation, and characterization of potential TGF — / 3 1 complexes from recombinant CHO cells are more fully described in the co-trial application filed on August 17, 1989, 07/353, 728. And for reference. 5.2 TGF — / 3 orange anti-cut as a pressure-applied coating Applicant found that r TGF — 々 1 can quickly and significantly lower blood pressure, indicating that BP regulation and / or hypotension may involve TGF — Lu. In this connection, through its ability to hinder the hypotensive effect of TGF- / 3, TGF-々citrus resistance can be used as a pressurization / decompression agent that can increase B P. Any composition that effectively antagonizes the hypotensive effect of TGF —> 9 can be used here, including but not limited to anti-TGF 8 antibody and TGF — / 3 receptor Ο For example, TGF — / 3 antagonists can be used to lose BP Under medical conditions, it is expected to increase BP to normal levels. This condition includes, for example, shock due to loss of blood volume, the risk of cardiomyopathy, and acute renal failure (please read the precautions on the back before filling this page). Binding · Order_ Line. Η 軍捃 m (CNS) Ping 4 specifications (210x297 gong) -15-205005 A 6 B6 Printed by the Beigong Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economy V. Description of invention (14) Exhausted hypotension. TGF_ / 3 can be administered alone or in combination with other compressive / decompression substances such as dopamine, epinephrine, aminophylline, etc. and / or co-administer. A complex containing an effective dose of TGF- > 3 antagonist with an appropriate pharmaceutical carrier can be administered to patients with hypotension or symptoms accompanied by hypotension via any appropriate route (including but not limited to injection, infusion and selective introduction) Improve BP. _In addition, TGF-/ 3 antagonists can be linked to carriers or targeting targets and / or incorporated into liposomes, withdrawal capsules, and controlled-release formulations before the snakehead drug. 5,2.1 Anti-TGF — θ Hangye Antibody that can inhibit the hypotensive effect of T G F — / 3 can be used as a pressurizer. Various methods known in the art can be used to produce multiple strains of antibodies against TGF-3 antigen-determined sites. In terms of antibody production, various host animals can be injected with TGF — / 3, or synthetic TGF — / S peptides for immunization, including but not limited to rabbits, mice, and rats. Various adjuvants can be used to increase the immune response, depending on the host type, including but not limited to Freund's (complete and incomplete), inorganic gums such as aluminum hydroxide, surface active substances such as lyso-lecithin, pluronic polyols, polyanions , P-eotide, oil emulsion, lyiBpet blood blue, dinitrophenol, and potentially available human adjuvants such as BCG (Bacille Calmette-Guerin) and Corynebacterium parvum. Monoclonal antibodies directed against the TGF / 3 epitope can be prepared by any technique used to produce monoclonal antibodies by culturing entangled cell strips. This includes but is not limited to Kohler and Milstein (1975, Nature 256, 495-497 (please read the precautions on the back before filling in this page) too earthworm ifif m Ψ03 63¾: quench miCNS) flat 4 media (210x297 public goods ) 161 205005 Printed by the Beigong Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 5. The hybridoma technology described in (15)), and the most recent human B-cell hybridoma technology (K-osbor eta 1., 1983 , Immunology Today 4:72) and EBV—Hybrid tumor technology (C o 1 eeta 1., 19 8 5, Μ ο η oc 1 ο na 1 A ntib o-dies and Cancer Therapy. Alan R · Liss, Inc., pp. 77-96). Antibody fragments containing deportable molecules are produced by known techniques. For example, such fragments include but are not limited to F (ab /) 2 fragments, which are produced by the hydrolysis of antibody molecules by pepsin, and Fab / fragments produced by reducing the disulfide bonds of F (ab>) 2 fragments, and can be used to Papain and reducing agents treat the two F ab fragments produced by the antibody molecule. The production of anti-TGF-Lu antibody was described in the co-examination application 7/353, 728, which was applied on August 17, 1989, and the co-examination application, 07/446, 020 Ο 5.2 applied on December 5, 1989 5.2 .2 TGF-/ 3 exaggerated exogenous TGF — / 3 receptor molecule can be a useful pressurizing agent, because of its cyclic TGF_; S and / or foreign endogenous receptors, it induces low TGF_y9 Blood pressure effect. TGF — / 3 can be prepared by the method described in the co-examination application 269, 524 filed on November 11, 1988. This case is fully incorporated herein by reference. 6. Tube administration: parenteral administration of r TGF-θ 1 to the blood limit of only monkeys (please read the precautions on the back and then fill out this page) Pack-line · Taifan Luo Km towel because of the house said (CNS) V4 Teague (210 to 297) 1. 17 — 205005 Printed by the Ordnance Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (16) The following are used for intravenous infusion to marmosets every day (Macaca f aso i-cularis) Part of the study to evaluate the toxic effect of r TGF — / 3 1 The results described here show that r TGF-/ 3 1 has a profound research effect on blood pressure. 6.1 Method steps ..... 6.1.1 Cat monkey Detect the blood pressure change of one mature male and two mature female marmosets from the daily TGF-Zhao 1 treatment. Monkeys feed on commercially available food and give water at will. Blood pressure was measured by a long-term arterial catheter implanted at least 5 days before the start of the study. One of the female monkeys (39-181) underwent general anesthesia and implanted a long-term venous catheter in the right epiphyseal vein. Another female monkey (2 9-8 2 5) and male monkey (B-3 44) also implanted long-term arterial and venous catheters in the iliac arteries and veins. The catheter is extended through an IV tube for intravenous injection to facilitate administration of the test article or vehicle control. 6.1.2 Test product formulation The test and / or control product formulation is completed daily before administration. Each 10mi of test article administration solution (0.0213mg / min by mixing 0. 66mi rTGF- / 31 mother liquor (0.46mg / mi? In 5mM HC) 2) and 9. 34m) 2 in PBS solution Ο. 1% monkey serum protein. The pH of the dosing solution was recorded every day after preparation and before administration. Calculate the dose based on the weight of the most recent unintubated tube and approximately the nearest 0.1 m i ?. Too recognized Zhang K due to iliSmfCNSVP / l hanging grid (210X297 public stoneware) _ jg _ (please read the precautions on the back before filling out this page) 装-线 · Printed by Beigongxiaof Cooperative, Central Standards Bureau, Ministry of Economic Affairs 205005 A6 __B_6_ V. Description of the invention (17) Recombinant mature TGF — / 3 1 The supernatant of the culture of marmoset ovary cells infected with complete human ape TGF- / 3 1 precursor coding sequence is purified and purified, as in 1989 The co-examination application filed on August 17 is described in 07/353, 728, which is fully incorporated herein by reference. 6.1.3 Lian Li Monkey 39—181 receives 1% monkey protein (excipient control group) in a volume of 2 / kg in PBS for 5 days. Monkeys 69-168 were treated with Burmese for 5 days at a dose of 0.17 mg / kg and a concentration of 0.0213 mg / m. rTGF — / 31 was given to monkeys 29-825, 39-181 and B344 at a dose of 0.51 mg / kg and a concentration of 0.0639 mg / m. Monkey 29_829 was treated for 3 consecutive days, monkey 39-181 for 3 consecutive days, and monkey B — 344 for 1 day. The body weight used to calculate the dose of the test article or the control group is the most recent body weight obtained from the unburied device. r TGF —々1 or excipient control group is injected intravenously via a long-term venous tube at 1.60 m $ administration solution / min through an input pump (Harvard Appara-tuS) and corrected by the sham operation method prepared by the test . Before giving the test article and the control substance, the catheter is washed regularly with 0.9% sterilized saline to maintain the visibility of the catheter. Record the volume, time, and date of r TGF — / 3 1 and the control group. 6.1.4 Rfn measurement of blood pressure measurement by long-term arterial tube by monkey 29—825, 39— Taimeng Rian side ΨIn Yinluan Deyi-19-(please read the precautions on the back before filling this page) Pack. Line < 205005 A 6 B 6 V. Description of the invention (18) 1 8 1 and B344 records. Blood pressure measurement is held for at least 1 minute before and after administration of the test article. The blood pressure measurement is also recorded as the revision or as directed by the study guide, if the measurement is clinically relevant. 6. 2 Linqing Observation Every day, observe the clinical abnormalities of apes, food and water intake, temperature, respiration rate, blood pressure, and other variables on the 29th. In addition, blood samples collected from each monkey were used as samples for hematology, serum chemistry, and immunoassay. 6.2.1 Effect of 1 ^ Ding 0/31/31 on 8 / B Two of the three monkeys who received t TGF — / 3 1 injection showed appropriate, obvious and sustained arterial blood pressure reduction. The third experimental monkey also had BP loss, but this result is somewhat difficult to judge in terms of the extremely low blood pressure of this animal before administration. Obvious BP fluctuations were also observed in control monkeys. The individual B P observations of these three experimental monkeys are shown in Table I. • ·························· (Please read the precautions on the back before filling this page ) Printed too cold iA Ψ 因 03¾: Pegging (CNS) A4 specification (210x297 public baking) -20-205005 A6 B6 V. Invention description (19) Table ul to TGF-jg 1 treatment: > Nanbei ifn of male and female marmoset monkeys pressed a loop g animal number sex dose (mg / kg) study arterial blood pressure at meal (mm Hg) 39-181

F 29-825F 29-825

F 0.51 B-344 0.51 經濟部中央標準局貝工消費合作社印製 0.51 1(前） 44 1(後） 48 1(4 Hr) 50 2(前） 60 2(後） 32 2(4 Hr) 46 3(前） 40 3(後） 32 3(4 Hr) 12 4 18 5 32 1(前） 108 1(後） 80 1(4 Hr) 48 2(前） 76 2(後） 76 3(前） 20 4 52 5 50 1(前） 104 1(後） 60 1(4 Hr) 36 2 20 3 32 4 22 5 20 9 36 (請先閲讀背面之注意事項再填寫本頁)F 0.51 B-344 0.51 Printed by Beigong Consumer Cooperative of Central Bureau of Standards of the Ministry of Economic Affairs 0.51 1 (front) 44 1 (rear) 48 1 (4 Hr) 50 2 (front) 60 2 (rear) 32 2 (4 Hr) 46 3 (front) 40 3 (rear) 32 3 (4 Hr) 12 4 18 5 32 1 (front) 108 1 (rear) 80 1 (4 Hr) 48 2 (front) 76 2 (rear) 76 3 (front) 20 4 52 5 50 1 (front) 104 1 (rear) 60 1 (4 Hr) 36 2 20 3 32 4 22 5 20 9 36 (please read the notes on the back before filling this page)

T 太蚯珥R泞LS丨fl Ψ K团MM报格（210x29?公货） -21 ~ 經濟部屮央橾準局兵工消费合作社印製 205005 A6 _ B6_ 五、發明説明（20) 猿猴29 — 825及B — 344各具適度（給藥後1 小時）之26%及42%BP減低。處理4小時後，猿痕 29—825之血壓降55%且猿猴B—344為65% 。原始BP降在後缠處理日中仍持缠，導致低血壓及休克。遠猴39—181對初TGF — /3處理（1日）無BP 降低之反應，可能因其預存之低血壓。令人感興趣的意，在處理前2日察得稍提高之BP,之後則察得持缠之BP 降。除在三隻經處理動物皆察得之戯刺性BP降及伴隨之休克/低血壓外，其他所察得歸因於TGF — /3 1之效果包括血液變化（紅血球，淋巴球及血小板減少）及免疫折衷方案（淋巴球對有絲分裂原之反應度降低）。此外，所有經處理動物無或只有最小食慾，且在處理期間皆無活力。猿猴在第4日艄下，並在第4及5日需流體治療，並在第5日因其垂死狀況而給予安樂死。猿猴39 — 1 8 1在第3及日4具黑血，在第8日産生敗血症，並在第8曰因其受損狀況而給予安樂死。猿猴B—344在第6至29 日皆正常。本發明不限於所示細胞糸，TGF — yS分子及檢定之範圍，其僅用於說明之部分。事實上，除本文所示者外，熟習此技藝者可由以上描述及附圖明白發明之各種改良。此類改良亦為本發明之範畴。 (請先閱讀背面之注意事項再填窝本頁) 太极误尺疳ΐΛ用+因囷S:JsmiCNS)甲4規格（210x297公帑） -22 - 2〇5〇〇5 S/-3〕°v 13〕°3v0i°9591〕Vi3si 313313 9°°Vi°0i30l0033°i3 91392 9i°°s:nl0ms00iv 51-*1J*1^01-0·【-Λ naj ttJl 391 n4*-1ou°-na-*=91 na-*0*-°-n·*-*3^1 WJV 3--1^【g J·*0OJJ 0 ^ °」d、l°—13- - -- ! 〇T 駿-< .............................................3J.....................................V.....19..... %iiii〕 ΛΧ ......................0.....0..................................................................... _<·< 001 - 3vi<_l3i;n3oos3i:J〕〕ui3<5i33-3sis33i3ii333soi35353i3333i3o3vvoois33odv3svoi3vivs9io33iiiv 0y^ ..............0.............i. ·3<ο·3ν·3----3ii ·1333. .V· :V33. :3:3313^Y B6I- iisovi3i53:Jsi3i353visi33iLJis!-3iv3vv-!sssi33v333i3i3is3sv sz-0S/ SI 〕0〕°iv ΰν°3°iv〕s 35 S3 3531391335 3iV 33 3 δαν4*-ϋ iu041-rvslswv SAl-Λ s-'nGwaxttsv·- "IV sr u> u卜 m «c。 C9 >»<· 〕9133ν 331 VI〕v°0y0s〕300〕3 sxu Jf ts 2-J-9*e【v,0lv0上 -T 太華珥 R 泞 LS 丨 fl Ψ K group MM report (210x29? Public goods) -21 ~ Printed by the Ministry of Economic Affairs, Pyongyang Bureau of Ordnance Industry and Consumer Cooperatives 205005 A6 _ B6_ V. Description of the invention (20) Ape 29 — 825 and B — 344 each had moderate (1 hour after dosing) 26% and 42% BP reduction. After 4 hours of treatment, the blood pressure of Ape Scar 29-825 dropped by 55% and Ape B-344 was 65%. The original BP is still entangled in the post-entanglement treatment day, resulting in hypotension and shock. Yuanhou 39-181 did not respond to the initial TGF / 3 treatment (1 day) without BP reduction, possibly due to its pre-existing hypotension. Interestingly, the BP that was slightly improved was observed 2 days before the treatment, and then the BP that was held around was lowered. In addition to the dramatic BP drop observed in the three treated animals and the accompanying shock / hypotension, the other effects attributed to TGF — / 3 1 include blood changes (red blood cells, lymphocytes, and thrombocytopenia) ) And immunocompromise (the lymphocyte response to mitogens is reduced). In addition, all treated animals had no or minimal appetite, and were inactive during the treatment period. The apes were sterned on the 4th day, and required fluid therapy on the 4th and 5th days, and were given euthanasia on the 5th day due to their dying condition. Ape 39 — 1 8 1 had 4 black blood on the 3rd and the 8th day, developed sepsis on the 8th day, and was euthanized on the 8th day due to its damaged condition. Apes B-344 were normal on days 6 to 29. The present invention is not limited to the range of cells, TGF-yS molecules and assays shown, it is only used for illustration. In fact, in addition to those shown here, those skilled in the art can understand various improvements of the invention from the above description and the accompanying drawings. Such improvements are also within the scope of the invention. (Please read the precautions on the back before filling the nest page) Tai Chi Misrule Use + Intrinsic S: JsmiCNS) A 4 specifications (210x297 public money) -22-2〇5〇〇5 S / -3] ° v 13 〔° 3v0i ° 9591〕 Vi3si 313313 9 °° Vi ° 0i30l0033 ° i3 91392 9i °° s: nl0ms00iv 51- * 1J * 1 ^ 01-0 · 【-Λ naj ttJl 391 n4 * -1ou ° -na- * = 91 na- * 0 *-° -n · *-* 3 ^ 1 WJV 3--1 ^ 【g J · * 0OJJ 0 ^ ° 」d 、 l ° —13---! 〇T 骏-< ....................................... 3J ... ................................. V ..... 19 .....% iiii) ΛΧ ...................... 0 .... .0 ................................................ ..................... _ < · < 001-3vi <_l3i; n3oos3i: J〕] ui3 < 5i33-3sis33i3ii333soi35353i3333i3o3vvoois33odv3svoi3vivs9io33iiiv 0y ^ ...... ........ 0 ............. i. · 3 < ο · 3ν · 3 ---- 3ii · 1333. .V ·: V33.: 3: 3313 ^ Y B6I- iisovi3i53: Jsi3i353visi33iLJis! -3iv3vv-! Sssi33v333i3i3is3sv sz-0S / SI〕 0] ° iv ΰν ° 3 ° iv) s 35 S3 353 1391335 3iV 33 3 δαν4 * -ϋ iu041-rvslswv SAl-Λ s-'nGwaxttsv ·-" IV sr u > ubm «c. C9 >» < · 〔9133ν 331 VI〕 v ° 0y0s〕 300〕 3 sxu Jf ts 2-J-9 * e [v, 0lv0 up-

U CJ a. uU CJ a. U

3 CD 4) ►-3 CD 4) ►-

O o o i. o a. o >>〇产〇O o o i. O a. O > > 〇 Production 〇

Ο O o o 〇Ο O o o 〇

卜 »- LJ ο. o o o U CJ a. t_)卜 »-LJ ο. O o o U CJ a. T_)

·-» 〇 m H-> o c o t5 CJ u o 4i C9 \Λ <.·-»〇 m H- > o c o t5 CJ u o 4i C9 \ Λ <.

〇 O u o a. o o u t. o a. o〇 O u o a. O o u t. O a. O

i. o 49 O «/) < «I卜 J Ui. o 49 O «/) <« I 卜 J U

σ»〇 U (S < o 3 0 •卜 _l o Μ O >»< -I < t. CJ «I o 的卜σ »〇 U (S < o 3 0 • Bu _l o Μ O >» < -I < t. CJ «I o 的卜

3 0 •J卜 J CJ *» 〇3 0 • J Bu J CJ * »〇

9 O r—式〇〇9 O r—Form 〇〇

〇 Ο I u o a. o 3 < ' #- < 〇 C9〇 Ο I u o a. O 3 <'#-< 〇 C9

O U U O a. o a o »- < C9 C3 moo 0)U o a. u 3 O 'O U U O a. O a o »-< C9 C3 moo 0) U o a. U 3 O '

〇 O « o · »— CJ < t9 L. t- «I o (/) < s o »- < « u> *— u < o *— 〇衊卜 > o σι〇 ft. o〇 O «o ·» — CJ < t9 L. t- «I o (/) < s o»-< «u > * — u < o * — 〇卜卜 > o σι〇 ft. O

< CJ 0. u> M < < o< CJ 0. u > M < < o

O CTtCJ CO L. C9 < 〇 u o -C 〇 I- < 1. Ci 4» O </) < C O y% *C < < i. LJ >»< Η- I— a o 邋»卜 __i 〇 4>卜O CTtCJ CO L. C9 < 〇uo -C 〇I- < 1. Ci 4 »O < /) < CO y% * C < < i. LJ >» < Η- I— ao sloppy »Bu__i 〇4 > Bu

_J CJ »— o <〇 ►-> cs (Q CJ ·— l_)_J CJ »— o < 〇 ►- > cs (Q CJ · — l_)

«Λ CS x< <«Λ CS x < <

4< O fr：4 < O fr:

vt O >t< J <vt O > t < J <

CX U < o O o fvrf ·— h- a «c -C o H- < a < 产< 〇 o r- 〇 <β ►—CX U < o O o fvrf · — h- a «c -C o H- < a < production < 〇 o r- 〇 < β ►—

> O W〇 ❸卜 Σ < a <> O W〇 ❸ 卜 Σ < a <

ti h-J CJ »— o 縐卜 > o O cncj 一 u C9 -"·<!> L. CJ -C o 卜*< ·— 〇 <〇 t— > C9 3 0 o o vi O >>< «J << H* »-O.U>ti hJ CJ »— o crepe > o O cncj a u C9-" · <! > L. CJ -C o 卜 * < · — 〇 < 〇t— > C9 3 0 oo vi O > > < «J < < H *» -O.U >

VI < «Ο O r— CJVI < «Ο O r— CJ

4>卜 J U 3 C9 0» — r- O <〇 t— 〇 o *— t. o a. o 3 < 〇 o 〇*>〇♦— V u 〇 ·-·〇.〇 <a ►— > 〇 « <，4 > Bu JU 3 C9 0 »— r- O < 〇t— 〇o * — t. O a. O 3 < 〇o 〇 * > 〇 ♦ — V u 〇 ·-· 〇.〇 < a ►— > 〇 «<,

r- O «< o 3 < o o u o < or- O «< o 3 < o o u o < o

3 CJ 4J I-J U 3 O ·— 〇 o u < U1卜 L. < •C CJ t— < •Λ < < < cn φ cj m «c卜 4/ o jc卜〇卜 w 〇 •i卜 X < 卜卜 «/ < ·— ►- L· U 41 Ο C/l <3 CJ 4J IJ U 3 O · — 〇ou < U1 Bu L. < • C CJ t— < • Λ < < < cn φ cj m «c Bu 4 / o jc Bu〇 Bu w 〇 • i Bu X < Bu Bu «/ < · — ►- L · U 41 Ο C / l <

•C ♦— I • 〇 W1 < x<• C ♦ — I • 〇 W1 < x <

J <X c 〇 —< 〇 o i. CJ 卜卜 \ 3 0 • 41卜J < X c 〇 — < 〇 o i. CJ 卜卜 \ 3 0 • 41 卜

• -J LJ• -J LJ

3 O <J C3 ·— O <β ►- > o 卜< X u> C O «- <3 O < J C3 · — O < β ►- > o BU < X u > C O «-<

〇 CJ a cs *— < 〇 C3 r— 〇 « »- > o〇 CJ a cs * — < 〇 C3 r— 〇 «»-> o

«Λ «C >.«c —} «c a «C 41 h-«Λ« C >. «C —}« c a «C 41 h-

*-· _J l_J σχ〇 u 〇 < < σι I I ( U I f c < » » ·*-· _J l_J σχ〇 u 〇 < < σι I I (U I f c < »» ·

3 O 4) H--JO 3 0 似卜"3 O 4) H--JO 3 0 似卜 "

_J O CT»f~_J O CT »f ~

U (J < o 3 0 4J h--J o 3 O ·— *i O <3 «« »— o < o 〇>〇 L. 〇U (J < o 3 0 4J h--J o 3 O · — * i O < 3 «« »— o < o 〇 > 〇 L. 〇

< LJ L. O O 〇o >— 盤駿 A-n rrr^bs « 205005 m cz® m il< LJ L. O O 〇o > — Pan Jun A-n rrr ^ bs «205005 m cz® m il

SI szlSI szl

OOCI v> <C < 〇 ❸►- JZ »— a- ^ L· — «I O </!卜 3 〇， «I卜 m 〇.〇 Oi L· is h- 3 O o o o o « k o a. kj wo v CJ 4Λ卜OOCI v > < C < 〇❸►- JZ »— a- ^ L · —« IO < /! 卜 3 〇, «I 卜 m 〇.〇Oi L · is h- 3 O oooo« ko a . kj wo v CJ 4Λ 卜

M (Λ < < < < IM (Λ < < < < I

<U C9 4Λ < o o U CJ a. cj «a 〇 < r- O < o 3 13 4J »--J LJ 9 U Φ »- —I LJ< U C9 4Λ < o o U CJ a. cj «a 〇 < r- O < o 3 13 4J» --J LJ 9 U Φ »-—I LJ

09 1. 〇 *-· < CJ < *- —J CJ L. CJ ^>v< 〇n3100vs usossv 13V 3ii°s〕vv〕iv3v0°i9 V2°13 5V3VV 15 vvvusvus 131 JMiilvaaldsvxl0J^i^Md^lsesv^uttsv-ACGSlJieiGSV s-Jastlsv sx〕 ϊζ ^ gi ..........................................i................................ ΰι 391:5333333113 3S ill〕S°°V3 iivvv3000v0°3s:}0v 3ii siDVDSSDSiiDVS'-'uv'-'l0 」as SX3SIH-V ts nal6J<4,f>',_0n,-0a::nG>'t0>'Gc,,JV ts 21dji UGSJV-A-A^-JMi-A 02Z 2~ 〕〕1 3°vv3°i3 ivuv3 Dssvs0913°〕3ΰν〕u°°iv 3i0il3°i3 3U 133 S3〕vv 9iv°9°i5 iiV ts ts lu°2155 usilvrv alsslojd ϋ iv SI nalnalaf OJdBi 5V iw^o-H'I OZZ S3 u o · f— H- · *— 〇 ♦ C3 O · >,〇 « *— i3 . O t3 . Pro CCC o · < · LJ . O . O · O · O · o · CJ · < o o · o . u · o . o · o · CJ · CJ * CJ · o · CJ · o · o . KJ . CJ · CJ , CJ · LJ · CJ卜 o · o · Leu CTG Leu CTG ..C Pro CCG *Λ O >>< -J < Gin CAG Cys TGC Asn AAC C7t〇 L. O < O 90 · u 〇 · >*-> 和O O 〇 « 4Γ • O' • »— o · 钃卜· · > o · c <v. r· Asn AAC u* r* Gin CAG G 卜 r· —O ，卜 > O 〇 ^ O . >%〇 · 。卜 · <〇 o · »— Ci · <.o . Asn AAC t. O 卜卜 «Λ CJ . >>ta · 1- « His CAT L· · >>< · 卜卜 · k. CJ >»< 卜卜 c «-> · *A < · «c < * wo · >>< · 卜卜 · 3 0 · V »- · O » —o 40卜 > o «Λ O · >t< · -J < · #— 19 〇 (£ · « CJ · —<J · < o . 41 (-1 产►· 一 < < · o · 卜 · 3 O . o u · «A o · X · J < « Leu CTG Pro CCC Ser AGC Thr ACG Pro CCC ^ 〇 · m »— ♦ > o . Leu CTG *A CJ · >>〇 · CJ卜 · Ser TCC Glu GAG y% 〇 · >»< · -J < · Pro CCA ..G wi < 〇 >>< _ —1 < · Ser AGC His CAC 1. o · 〇 o · t/l < , Glu GAG ΙΛ < ♦ >>10 · CJ卜 ♦ Phe TTC O · —· —< · &. o · · Leu CTG Ser TCC * «* • cc • fs VI 〇 · >»〇 · cl卜 · c 严 0.0 · L. C9 · u- r* Gin CAG C VC r" Ale GCG ir oc n σ»ο · U LD · < o · k> CJ 卜 >»< · 卜卜 · VI C9 · >>< · < · u o · « CJ . H- < · Gin CAG o · <e I- · > C9 · c o · < · < < , 0.0 · L. O · 0.0 . < o · Pro CCG 4/ CJ · r— ►- ♦ —< . Thr ACC · f- O · o o · LCU CTG —o <〇卜 > C9 Met ATG 0.0 * yt «C ♦ < C9 · Leu CTC Ser AGC •Λ U» · o卜 Asn AAC 3 0 · υ i- · J CJ · As p GAC CL· 〇 . U t3 · y% CJ XC3 CJ卜 Ser TCC to o ♦ «— o . < o · M o >»< · 4t h~ · 一 < · 〇〇 L. O a. cj Leu CTG 〇*<. · u 〇 . <.<-)· 〇1〇 · t. 〇 . < o · &. o . >^< · ►— Η* 礴〇 r- O < 〇 Gin CAG OIO · i. o . < o · o o ♦ It : Pro CCC <〇 o o < o Glu GAG V% CJ · —< · X CJ · 0.0 · < · < · vl H- <-* . O H-. L. C9 Φ CJ \Λ h- 〇 · <〇卜，· >► c? · σ*ο · i. 〇 · < o · fti H- · 产卜 · —< · o o ♦ &. o . a, o . « O CJ < O v% C9 · <C . .....................V.......u.........0----0----v..........v^vvvi 12:0:^3991:3:13:^953:^3313:5^5535111^1.91309939.1133331131313333333533:^33:}000000000000 i #r J®一缺 ¾ ^ m 205005 5/.CJUVooloCJv laiaiCJoovo13 08arju3voio313i-s OLDolvoiDt-u lououvoloaiD iol 3ViCJvoaJ.V135 5^.^11.35 3-1^0)5 31^3 15 114115 33-1012 3-141113-1351:3-^5 5-)15(0^3.^,1,52 1¾ 02 - -- ------------------- --- -----------T VVVVVLLLLUo3o.LLLLLLU3vsvvvvvl.LLi35.L3LLV.LLLLLLLL3LLLVuii3.UVVD3.LLLovlvl3.LvaLX3aL313v5vv33vvv001 · vvsvsvvuvsvvvvv3ivCJutJ.LU,3vvvsaLLLauvDvlosx.LSoouiluJJX.LUlo3vl5t3voaVS.LLVVOOVVV.LLLLiu{-.ulo3066l - 3iuvl3vsuiJ.UOS3.Luvlvv.LLVluyulayooi3iuy.LLoou51DVVS381(-)vilo.LL3.LLLLLU.VVovs31yilv3i3i3o3o 86f\l- ,LLU.Ui31u.Lo3t-3s3Dvlvv.uoV.LoillviilvvvJJ.u18x3u113sCJ13vslu5v3si3ss3335333io«x33a;Di3511ov·」 z.6?CJ33s3VQOyo3iliDo3ix3ioisli.LL333oo33yusa3133vo83f-iD313yoloCJa33 i9tj — oic 〇〇 ε § ics §9 38oviuvi 3S vvoos 33V 3UV 3vo333avou3i 333 33US3 sv suuov S3 9VV Dvoovu3X3 3X1 33 <eIV J>>1 J>.1 nG 33asv JuioMJV nG 3J«nGn>>uES<3IV civ MJVeaJVL.w«0«3ηλι nG UGnansq d5 lr>6 ςβ uuv 33 VuuvuvvCJuolvoul 150100V0000000U13 so 3338 3S iuCJicci 33VS 50333 ιανυυν eoJV JJCl ts U5 J>»i 3U t 的 3二 T5 fuCJOJCL.OJjt5 nG nG oinIuOJa.J>»i d5 nG OJa.0Ja.J3toJMl • 0卜.09 uluovvoloovvoavolDalvovaooooooolv DtJoosulvoov DVV 38U1V 3X1O3CJVO01V1V0 3¾ 5V π«_]Μλ_ιηνη»ιΛη Jws na-IoucToiobaJV aHfgs nG4)11 31.V sfrMJV 13Σ3£ ulodsv qaxdsv nal-lJlu Sfls iuluouvav 5 V llollovouuululoiuv jas OJ0.JM1 JMi 15 15 £d εχοΙΒΛ JMH SI , 3U 10V33303001Vouu aMauJMX OJa.OJa.au 52 V503108311 ·」11383Β31νο5 «IV VVVCJvHlluovuovv Π3 Jaco2d «£0.«£ OJaLOJa.4J3Ha.l;v 3TI nxl Jxl 15 ntocxl Oil09 1. 〇 *-· < CJ < < UO «IO 4/1 < 3 O 4 > *-—J CJ L. CJ ^ > v < 〇n < uo < o 0.0 uo 1. o tl O ～卜 c 卜 V »< -c · < co < < < uu 4« iD% / \ < ^ rkr% / '0 ............ .................................................. ° 3 .......... 3V〕〕 0 ° 13:> 3100vs usossv 13V 3ii ° s〕 vv〕 iv3v0 ° i9 V2 ° 13 5V3VV 15 vvvusvus 131 JMiilvaaldsvxl0J ^ i ^ Md ^ lsesv ^ uttsv- ACGSlJieiGSV s-Jastlsv sx〕 ϊζ ^ gi ......................................................... .i .......................... ΰι 391: 5333333113 3S ill〕 S °° V3 iivvv3000v0 ° 3s:} 0v 3ii siDVDSSDSiiDVS '-' uv '-' l0 '' as SX3SIH-V ts nal6J < 4, f > ', _ 0n, -0a :: nG >' t0 > 'Gc ,, JV ts 21dji UGSJV-AA ^ -JMi- A 02Z 2 ~〕〔1 3 ° vv3 ° i3 ivuv3 Dssvs0913 °〕 3ΰν〕 u °° iv 3i0il3 ° i3 3U 133 S3) vv 9iv ° 9 ° i5 iiV ts ts lu ° 2155 usilvrv alsslojd ϋ iv SI nalnalaf OJd ^ o-H'I OZZ S3 uo · f— H- · * — ○ ♦ C3 O · >, 〇 «* — i3. O t3. Pro CCC o · < · LJ. O. O · O · O · O · CJ · < oo · o. U · o. O · o · CJ · CJ * CJ · o · CJ · o · o. KJ. CJ · CJ, CJ · LJ · CJ Buo · o · Leu CTG Leu CTG ..C Pro CCG * Λ O > > < -J < Gin CAG Cys TGC Asn AAC C7t〇L. O < O 90 · u 〇 · > * -> and OO 〇 «4Γ • O '•» — o · 钃卜 · > o · c < v. r · Asn AAC u * r * Gin CAG G 卜 r · —O ，卜> O 〇 ^ O. &Gt;% 〇 ·. ·································································· · K. CJ > »< Bubu c«-> · * A < · «c < * wo · > > < · Bubu · 3 0 · V»-· O »—o 40 Bu > o «Λ O · > t < · -J < · # — 19 〇 (£ ·« CJ · — < J · < o. 41 (-1 production ► · one < < · O · Bu · 3 O. Ou · «A o · X · J <« Leu CTG Pro CCC Ser AGC Thr ACG Pro CCC ^ 〇 · m »— ♦ > o. Leu CTG * A CJ · > >; 〇 · CJ Bu · Ser TCC Glu GAG y% 〇 · > »< · -J < · Pro CCA ..G wi < 〇 > > < _ —1 < · Ser AGC His CAC 1. o · 〇o · t / l <, Glu GAG ΙΛ < ♦ > > 10 · CJ Bu ♦ Phe TTC O · — · — < · &. o · · Leu CTG Ser TCC * « * • cc • fs VI 〇 · > »〇 · cl bu · c Yan 0.0 · L. C9 · u- r * Gin CAG C VC r " Ale GCG ir oc n σ» ο U LD · < o · k > CJ BU > »< · BU · VI C9 · > > < · < · uo ·« CJ. H- < · Gin CAG o · < e I- · > C9 · co · < · &Lt; <, 0.0 · L. O · 0.0. &Lt; o · Pro CCG 4 / CJ · r— ►- ♦ — < .Thr ACC · f- O · oo · LCU CTG —o < 〇 BU > C9 Met ATG 0.0 * yt «C ♦ < C9 · Leu CTC Ser AGC • Λ U» · o BU Asn AAC 3 0 · υ i- · J CJ · As p GAC CL · 〇. U t3 · y % CJ XC3 CJ Bu Ser TCC to o «— o. &Lt; o · M o >» < · 4t h ~ · one < · 〇〇L. O a. Cj Leu CTG 〇 * <. · u 〇. <. <-) · 〇1〇 · t. 〇. < o · &. o. > ^ < · ►— Η * 礴〇r- O < 〇Gin CAG OIO · i. o. < o · oo It: Pro CCC < 〇oo < o Glu GAG V% CJ · — < · X CJ · 0.0 · < · < · vl H- <-*. O H-. L. C9 Φ CJ \ Λ h- 〇 · < 〇卜， · > ► c? · Σ * ο · i. 〇 · < o · fti H- · 产卜 · — < · oo ♦ &. o. a, o . «O CJ < O v% C9 · < C. ..................... V ....... u ..... .... 0 ---- 0 ---- v .......... v ^ vvvi 12: 0: ^ 3991: 3: 13: ^ 953: ^ 3313: 5 ^ 5535111 ^ 1.91309939 .1133331131313333333533: ^ 33:} 000000000000 i #r J®One missing ¾ ^ m 205005 5 / .CJUVooloCJv laiaiCJoovo13 08arju3voio313i-s OLDolvoiDt-u lououvoloaiD iol 3ViCJvoaJ.V135 5 ^. ^ 11.35 3-1 ^ 0) 15 114115 33-1012 3-141113-1351: 3- ^ 5 5-) 15 (0 ^ 3. ^, 1,52 1¾ 02------------------ --- --- ----------- T VVVVVLLLLUo3o.LLLLLLU3vsvvvvvl.LLi35.L3LLV.LLLLLLLL3LLLVuii3.UVVD3.LLLovlvl3.LvaLX3aL313v5vv33vvv001 · vvsvsvvuvsvvvvv3ivCJutJ.LU, 3vvvsaLLLauvDvlosx.LSoouiluJJX.LUlo3vl5t3voaVS.LLVVOOVVV.LLLLiu {-. Di3511ov · "z.6 CJ33s3VQOyo3iliDo3ix3ioisli; ulo3066l - - 3iuvl3vsuiJ.UOS3.Luvlvv.LLVluyulayooi3iuy.LLoou51DVVS381 () vilo.LL3.LLLLLU.VVovs31yilv3i3i3o3o 86f \ l-, LLU.Ui31u.Lo3t-3s3Dvlvv.uoV.LoillviilvvvJJ.u18x3u113sCJ13vslu5v3si3ss3335333io x33a«? .LL333oo33yusa3133vo83f-iD313yoloCJa33 i9tj — oic 〇〇ε § ics §9 38oviuvi 3S vvoos 33V 3UV 3vo333avou3i 333 33US 3 sv suuov S3 9VV Dvoovu3X3 3X1 33 < eIV J > > 1 J > .1 nG 33asv JuioMJV nG 3J «nGn > > uES < 3IV civ MJVeaJVL.w« 0 «3ηλι nG UGnansd 6 VuuvuvvCJuolvoul 150100V0000000U13 so 3338 3S iuCJicci 33VS 50333 ιανυυν eoJV JJCl ts U5 J > »i 3U t's 3 二 T5 fuCJOJCL.OJjt5 nG nG oinIuOJa.J >» I0JoJoVoJoVJV0VJ0V0VJ0VJ0 3X1O3CJVO01V1V0 3¾ 5V π «_] Μλ_ιηνη» ιΛη Jws na-IoucToiobaJV aHfgs nG4) 11 31.V sfrMJV 13Σ3 £ ulodsv qaxdsv nal-lJlu Sfls iuluouvav 5 Vlli OJa.OJa.au 52 V503108311 · "11383B31νο5« IV VVVCJvHlluovuovv Π3 Jaco2d «£ 0.« £ OJaLOJa.4J3Ha.l; v 3TI nxl Jxl 15 ntocxl Oil

Is .LLuuuoouxaLCJloulousCJui 33 SV0UHU0J.UJ.Luovooouiuvuluoo ds« n31J>.l><G SKo311lcs»5oJatnu3MJVS«n«>»3na_lJa</l>>I3I5Jas>.Io•ςει 0 i013 llyovuivlvluas ilv.uouVV3V5 luuDrJ 5V yosv VBDVVovoull Jbuli.Loio5vt_lixo5 S14;u CG J>>卜 n3_lnto4;二 MJVU5 n3 OJa.15eeJV eT«sc>.-JOJd CSV UG na_)boJV4>£ 15 -Jeov aMa-nG SI sool 0 53 VVV oluOLL iVV 3ui.LCJuIs ovvovoolv V3USH31U3V0 111 vouxio lHv 3V 31iuvHu33 vuv 3V1 Μτνσ>>>-)ΙΒΛ 5_lusv J3COBI«Scev S>.J nToqvH Hs Jvs ΙΒΛα,εν a£eoJV 15VIJboJV aMa-Jlu oi MJV JS-.10卜一 . 〇9一 - ®cvl缺 2〇5〇〇5 vs VXD VVD VVD l°vvvv 1VV ss Π3 na-'n-'onGJS^exlucv 0卜2 . OSL Viv vs 111 S3DJJ 3VVUDV wu sXi aqdxG S1USV3JV s- ssV3U°°V5 33 3V v°v VVV nI0>'TunTufl,5MJ<:JMXlo>>-J 0¾ 0°6Is .LLuuuoouxaLCJloulousCJui 33 SV0UHU0J.UJ.Luovooouiuvuluoo ds «n31J > .l > < G SKo311lcs» 5oJatnu3MJVS «n« > »3na_lJa < / l > I3I5I0I0 VBDVVovoull Jbuli.Loio5vt_lixo5 S14; u CG J > > Bu n3_lnto4; II MJVU5 n3 OJa.15eeJV eT «sc > .- JOJd CSV UG na_) boJV4 > £ 15 -Jeov aMa-nG SIVOL 0 VV LCJuIs ovvovoolv V3USH31U3V0 111 vouxio lHv 3V 31iuvHu33 vuv 3V1 Μτνσ > > >-) ΙΒΛ 5_lusv J3COBI «Scev S > .J nToqvH Hs Jvs ΙΒΛαJ aJJVJJVV 9 一-®cvl missing 2〇〇〇〇5 vs VXD VVD VVD l ° vvvv 1VV ss Π3 na-'n-'onGJS ^ exlucv 0 Bu 2. OSL Viv vs 111 S3DJJ 3VVUDV wu sXi aqdxG S1USV3JV s- ° ss °° V5 33 3V v ° v VVV nI0 > 'TunTufl, 5MJ <: JMXlo > > -J 0¾ 0 ° 6

Gsl seu 0°2l °HuiJbvvv t5 15 s- 3S VVV 1V3 ds**51==Gsl seu 0 ° 2l ° HuiJbvvv t5 15 s- 3S VVV 1V3 ds ** 51 ==

333¾°¾ 2dΦΤΙWII 3V1 Is 1VV 1°1 vyviom 13V J^HUCV USVJa^OJ41·Tr/vdx·1·Jm 092 33 ixosl vs J.V3 110 i°0 13 •-H naidjj, nGSS T5 H3dsv s y^Dsuiv 3V1 3UU°V3°3V 53 」品 dsv OH J-WJV UG JMi o」d °lz u°i 331 333 lai°V3 Vli 30V "xusx0OJ11·5^3^---30-1J^^ i 13V vl°iva3xl 33i 313001 JMX IB5*dsv 3 131 V3V V1X Is vvvu31°s USU3V OS V33XUVD liusvuvl 331333011-=1135 V13°l°ui0 13 V3UU3VUSU°D Iguvvsv c5 JMi u-UIQJa^nGS-'HJ^JXXt^o上 sls-lyvH nsnans-ls-IH 2d JMlsl-DJVSesvsx-1 ol£ vvvsv J,UV3U1°VV V1V luv sv 33 HVD iuu5v 33V 1U V3V 33i33V 300 Isilv 185D.LLL VDV 3° exnsj**Ji ja的"xnan JMlsxluGdcv^G JWSJqx Jx*-JMH α^έ,ι^ΙοαεναΙΙ^ΙΟΛΙν^Μίιθν^ιν 562 ^ i 113 να3103ίυ301 001 1VV 13 D3D1D 1VV svixxoDl 1? 3°D 0°0 13 Dll 18 SJOiMJVn^JexucK0U5 d"v CI3 ΧΛΛ USVWJVWMdM^UJ^l^IV-'Tvdcv S-JH-,V --S3- luuuvv uoo ouo 133 iuialiuvvu3ulvvuvluD0 VVV 33uv3°v3vlvuui vvv00iu°uVXD is DDVUVVUX1 1VD11V3V1 ^2^^331^ ucvelvcsv JKlATSSXIOinGCJH^uaJisxndJli0naldevMJV"xnaMd d53【I」λχ -0i£-。名- vyivv V1V33V Is 1V1 VI! 3uvuiu31uuDV0DV 3SU3 luv°s SI Hovsl Vll lvl°30u°l 3D 3u OJdu5 ou」M1 CSV J-nvnjas 51-5 SJVJVS S-*H U5 JqHdsv JSJa^dJl naqj^l OJCjs-EIVit3 I-S6£-^- 3V VVVODD ilv3vluvl 3X0 X1V33V Viui33v3 V1X 1VD ν503131ϋ301301 lylui 18131 S°V3 JMlwi^TDVULfJ^l 31^11 JU1 naqoinlo nalds ut°ts 15^^3532^^3^^31^:^1^33 02t?-〇U- ° 95EI VVVUUVUV30U<1UVVV3011UX1VV VV1 3°v 3ul VVV331 i3i〇vv viu-.iv°lv 1VV 131 110 U5 V3 llv°vv°y ^3^5^^-1 *J4,XU"<£J4,S SHUI0nsausxqoj*1· •s-^- /1必 j licsl贼 205005333¾ ° ¾ 2dΦΤΙWII 3V1 Is 1VV 1 ° 1 vyviom 13V J ^ HUCV USVJa ^ OJ41 · Tr / vdx · 1 · Jm 092 33 ixosl vs J.V3 110 i ° 0 13 • -H naidjj, nGSS T5 H3dsv sy ^ Dsuiv 3V1 3UU ° V3 ° 3V 53 ”Product dsv OH J-WJV UG JMi o” d ° lz u ° i 331 333 lai ° V3 Vli 30V " xusx0OJ11 · 5 ^ 3 ^ --- 30-1J ^^ i 13V vl ° iva3xl 33i 313001 JMX IB5 * dsv 3 131 V3V V1X Is vvvu31 ° s USU3V OS V33XUVD liusvuvl 331333011- = 1135 V13 ° l ° ui0 13 V3UU3VUSU ° D Iguvvsv c5 JMi u-UIQJa ^ nGS-ts ^ lyvH nsnans-ls-IH 2d JMlsl-DJVSesvsx-1 ol £ vvvsv J, UV3U1 ° VV V1V luv sv 33 HVD iuu5v 33V 1U V3V 33i33V 300 Isilv 185D.LLL VDV 3 ° exnsj ** JiGJ &G; lslsx JWSJqx Jx * -JMH α ^ έ, ι ^ ΙοαεναΙΙ ^ ΙΟΛΙν ^ Μίιθν ^ ιν 562 ^ i 113 να3103ίυ301 001 1VV 13 D3D1D 1VV svixxoDl 1? 3 ° D 0 ° 0 13 Dll 18 SJOiMJVn ^ VexJJVK3VJJVJV3 UJ ^ l ^ IV-'Tvdcv S-JH-, V --S3- luuuvv uoo ouo 133 iuialiuvvu3ulvvuvluD0 VVV 33uv3 ° v3vlvuui vvv00iu ° uVXD is DDVUVVUX1 1VD11V3V1 ^ 2 ^^ 331 ^ KlxJJJJJJXJJJJJJ ldevMJV " xnaMd d53 【I 」λχ -0i £-. Name-vyivv V1V33V Is 1V1 VI! 3uvuiu31uuDV0DV 3SU3 luv ° s SI Hovsl Vll lvl ° 30u ° l 3D 3u OJdu5 ou 」M1 CSV J-nvnjas 51-5 SJVJVS S- * H U5 JqHdsv JSJa ^ lJ NaJ I-S6 £-^-3V VVVODD ilv3vluvl 3X0 X1V33V Viui33v3 V1X 1VD ν503131ϋ301301 lylui 18131 S ° V3 JMlwi ^ TDVULfJ ^ l 31 ^ 11 JU1 naqoinlo nalds ut ° ts 15 ^^ 3532 ^^ 3 ^ 33 02t? -〇U- ° 95EI VVVUUVUV30U < 1UVVV3011UX1VV VV1 3 ° v 3ul VVV331 i3i〇vv viu-.iv ° lv 1VV 131 110 U5 V3 llv ° vv ° y ^ 3 ^ 5 ^^-1 * J4, XU " < £ J4, S SHUI0nsausxqoj * 1 · s-^-/ 1 must j licslthief 205005

Ills {v^odo3.LDSXE33vvvvlvv.LLDJ.3v3vvvvvs\mv3:l§mm3x votnos 13DV50vvluuDlo3slvl31vllulsv3vvvsl333vovvvl3VV.LL331313vv338slvlv.Llv.LLovulovvCJyvvvslvsu 9ln6l lvvlxvlvo130svvvvvv5voocls3vvv313vvo113vv8<3:v.u8oi-voo.LVCJlovvvosuDlllCJ«J:lvvsovuVDolDvvvlvisloas 9ln8l vvvvlvvvs3.uooo9.LXV3illouvv31VV.LVlvl3v.u.o131all<:.LLlv.u3slu3vvlsvDV.LL3xCJV.LLuxl3uuxluoovooovxvv3 glnAl0ivl0ollv5x3.LLLLlv3il3v3x3u33o0:>aivl81x030VIDovlDlu0lltJVDuJ.uVVJ.lv03;uvvvv033vvvuv3v3<svc_)v5v5v 9ΙΠ91 νονουννοννϋιοινιχννιχοχανιχνιϋννονδοιονανννινννννννονννϋαχαιχνννυοννοννυνανονονοιοννχοιυονιουν 9ITV91 3xilsvl3i.Lvil33uo<3:sxiCJvvvvvv3va51ul3oD.L3VVVV.LLox.uul:ulDloEuVSDEuvCJvos.u.ovlovloo3uDVVVS volntn lix.LL«3:v«c<cv<£u31o31S.LLS.U3ooVOVDVVlv3vvvvoVSVV.LD.LL81sisuvss3su3131slvvlsxvol51vv3voiv ®6· 001- 一丨 ςζ OS lIlls {v ^ odo3.LDSXE33vvvvlvv.LLDJ.3v3vvvvvs \ mv3: l§mm3x votnos 13DV50vvluuDlo3slvl31vllulsv3vvvsl333vovvvl3VV.LL331313vv338slvlv.Llv.LLovulovvCJyvvvslvsu 9ln6l lvvlxvlvo130svvvvvv5voocls3vvv313vvo113vv8 < 3: v.u8oi-voo.LVCJlovvvosuDlllCJ «J: lvvsovuVDolDvvvlvisloas 9ln8l vvvvlvvvs3.uooo9.LXV3illouvv31VV.LVlvl3v. u.o131all <:. LLlv.u3slu3vvlsvDV.LL3xCJV.LLuxl3uuxluoovooovxvv3 glnAl0ivl0ollv5x3.LLLLlv3il3v3x3u33o0: >aivl81x030VIDovlDlu0lltJVDuJ.uVVJ.lv03; uvvvv033vvvuv3v3 < svc_) v5v5v 9ΙΠ91 νονουννοννϋιοινιχννιχοχανιχνιϋννονδοιονανννινννννννονννϋαχαιχνννυοννοννυνανονονοιοννχοιυονιουν 9ITV91 3xilsvl3i.Lvil33uo < 3: sxiCJvvvvvv3va51ul3oD.L3VVVV.LLox.uul: ulDloEuVSDEuvCJvos.u .ovlovloo3uDVVVS volntn lix.LL «3: v« c < cv < £ u31o31S.LLS.U3ooVOVDVVlv3vvvvoVSVV.LD.LL81sisuvss3su3131slvvlsxvol51vv3voiv ®6 · 001- a

OOC s- § ls 2卜9 卜hi °t ou°13 iyvs ly V1D 5D V5 V03°t°s v°i V3V3VV-U 3J.luiv SI V1V°3VUS 5V 30V 33 S121I5 o.ldns oiicp HI**33 fv Sjn3 Jasti 5V aMaafiw 3-ts ςΞ JM1 ts UIU s ςει 0v**uxx°3031vi US V50vvu33uv vv°°10 D1V vx0OL000uyvulu0v0uvv0u:)·°ν1 3V1°3 3°3 "^IWMdcxldcvJxiiu3-·01125=1 jmi Π3 ΙΕΛ-ί slle/VWJVJMi-'HAnGsxl-v J^l Lxi αί:νΛ【ν οει oil 3νο03υννο3υοον0ουυονοο:>0luv030ooy0olD083vo00uyvuov:}vvuvl0.Gy0;u:}u10 y° nloointooJ^nG 2d Π3 5sts n3-ue3 JisJvdcvMJV LMX ts ccv sl-vnvnJVA e—v s- 0v038DXD000u00yu0yuiu0v0o000300vu0000000vs0ui300u°iD0vv yl°Guiv os°8 33 OJdnaJoi-OOJJOis/y nGXG UGtsoi oit*0·0-·*1nalwjv na-'fx-'Ja^na-'^UCGXG 01 s °8ulv bv 03°XV3S0VVU03°VVUXO 0100s0lv°3 35-3v0vvuniu°v 331 s.°33 33 ΰ°3ΰ 33 Λί Λί SJ 35 &2 ^£- wi ni ^£- ^-.0 m3 "ςε 3033¾ iyouvolo OL00i°vi3u0i OLDο1οο30νχοαιοοχ3οπυοχ3ο13ο8ο1ο°οο031uuu0y°iv sija^oijqx 3115 na-·s-ldjl s-'s-'OJJna-'s-'sloinalnalMJV 5-1^3 is oiojdtw U2 ^ UUOUOGUOOU0080000UOOHU<OOOOHOHOOUOOt.oHoaao?us^oo<oaooyoooooo80<aoHUUOOO<ooooo<ooosouo^t.^o 3ν1υο138ϋονο018υ1ί2ν0&ν3ο3χΰυ1υ1Ηοΰϋ:5αι330331υ3ΰχο:}ον3ο1ο3υ:χ}0:}3νο3ΧΓν13οο1ομ-μ1ν 1χοον131ν33381υ1ν03°ν,ιο01υυ105:>ίο1νϋνν!-3νοο318νυ810101νοο0ν s' °vi ννν0νυ3νχαιο°3 olDlsuvrJUVDOlQSV**!!0***1uio°uv JXICXJUIO JXH 2-13315"111 US n-,u-,5"><-J3-,c>'-Jn«,-JMJV 0-091 OHO OHO Hou ot 0<0 <ou oou OUH nannJWJV slnGelv^」**匕4140 3V〕〕0°lu°s l00v000lD°0uyv iuvulluDD JVVDivDVOOLDSUUlDSVDVV is VVV33V 33 i°l Jmes naldsvxGW'-VWJVXGJMX JMXVfx-10112-1^215=5 n^IJMXCSV dMv CAItsdcv cxus s UUH UUH 0<u ouo 00< tu oou hp uou 0<u 50 000 «00 000 00< ut· 00H 0<υ uuu οδ Fo <00 uo< 05 J»-的 βΛυςτΗ'"--'**ts nelMJVaMd-DnGa-nGXIQXGBJYJO^naldJl UGW·-**15 15"°Lux 15 OS SN 13 111 luiohto01ovo03u031uvvuovu3us00G0i30ou3vvuov3i3 3U 53 001 331 1VV3VV DOV dcvafjasnidjin-·0oj丄 J^^ucv .^^2,1-3 nwJS-'MJVCCV J°5 391.^1^^^1-^^=5 u"v J4*40 S6l£l οδI 20 51 lt>691 9? r-6:l8251 § ooOJl6二一tlsl696 s s auluav VSU13 IvoV5 3U0U3VCJS 3ios333vCJCJD alvoxo iloOLooxi 13308CJvvoiv 30315 11V Ja<oJals υ-Ηοπα)ηolJH U3BTV 抝 J<snG331 OJa.JMXSJH lvH πβ)η ηαιιπνη a£ OJdtlaJVcsv SHirasfHon 0卜2 S2OOC s- § ls 2 Bu 9 Bu hi ° t ou ° 13 iyvs ly V1D 5D V5 V03 ° t ° sv ° i V3V3VV-U 3J.luiv SI V1V ° 3VUS 5V 30V 33 S121I5 o.ldns oiicp HI ** 33 fv Sjn3 Jasti 5V aMaafiw 3-ts ςΞ JM1 ts UIU s ςει 0v ** uxx ° 3031vi US V50vvu33uv vv °° 10 D1V vx0OL000uyvulu0v0uvv0u:) ° ν1 3V1 ° 3 3 ° 3 " ^ IWMdcxld3 -ί slle / VWJVJMi-'HAnGsxl-v J ^ l Lxi αί: νΛ [ν οει oil 3νο03υννο3υοον0ουυονοο: > 0luv030ooy0olD083vo00uyvuov:} vvuvl0.Gy0s3 = y0; u: 3) ccv sl-vnvnJVA e—v s- 0v038DXD000u00yu0yuiu0v0o000300vu0000000vs0ui300u ° iD0vv yl ° Guiv os ° 8 33 OJdnaJoi-OOJJOis / y nGXG UGtsoi oit * 0 · 0- · * 1nalwjv na-G'J'na- '' ° 8ulv bv 03 ° XV3S0VVU03 ° VVUXO 0100s0lv ° 3 35-3v0vvuniuni ° v 331 s. ° 33 33 ΰ ° 3ΰ 33 Λί Λί SJ 35 & 2 ^ £-wi ni ^ £-^-. 0 m3 " ς ε 3033¾ iyouvolo OL00i ° vi3u0i OLDο1οο30νχοαιοοχ3οπυοχ3ο13ο8ο1ο ° οο031uuu0y ° iv sija ^ oijqx 3115 na- · s-ldjl s-'s-'OJJna-'s- ' sloinalnalMJV 5-1 ^ 3 is oiojdtw U2 ^ UUOUOGUOOU0080000UOOHU < OOOOHOHOOUOOt.oHoaao us ^ oo < oaooyoooooo80 < aoHUUOOO < ooooo < ooosouo ^ t ^ o 3ν1υο138ϋονο018υ1ί2ν0 & ν3ο3χΰυ1υ1Ηοΰϋ:?. 5αι330331υ3ΰχο:} ον3ο1ο3υ: χ} 0:} 3νο3ΧΓν13οο1ομ-μ1ν 1χοον131ν33381υ1ν03 ° ν, ιο01υυ105: > ίο1νϋνν! -3νοο318νυ810101νοο0ν s' ° vi ννν0νυ3νχαιο ° 3 olDlsuvrJUVDOlQSV ** !! 0 *** 1uio ° uv JXICXJUIO JXH 2-13315 > 111 US &; -J3-, c > '-Jn «,-JMJV 0-091 OHO OHO Hou ot 0 < 0 < ou oou OUH nannJWJV slnGelv ^' '** dagger 4140 3V〕] 0 ° lu ° s l00v000lD ° 0uyv iuvulluDD JVVDivDVOOLDVUlV VVV33V 33 i ° l Jmes naldsvxGW'-VWJVXGJMX JMXVfx-10112-1 ^ 215 = 5 n ^ IJMXCSV dMv CAItsdcv cxus s UUH UUH 0 < u ouo 00 < tu oou hp uou 0 < u 50 000 < 00 00 < 00H 0 < υ uuu οδ Fo < 00 uo < 05 J »-of βΛυςτΗ '"-' ** ts nelMJVaMd-DnGa-nGXIQXGBJYJO ^ naldJl UGW ·-** 15 15 " ° Lux 15 OS SN 13 111 luiohto01ovo03u03 vu3us00G0i30ou3vvuov3i3 3U 53 001 331 1VV3VV DOV dcvafjasnidjin- · 0oj 丄 J ^^ ucv. ^^ 2,1-3 nwJS-'MJVCCV J ° 5 391. ^ 1 ^^^ 1-^^ = 5 u " v J4 * 40 S6l £ l οδI 20 51 lt > 691 9? R-6: l8251 § ooOJl6 21 tlsl696 ss auluav VSU13 IvoV5 3U0U3VCJS 3ios333vCJCJD alvoxo iloOLooxi 13308CJvvoiv 30315 11V Ja < oJals3) ) η ηαιιπνη a £ OJdtlaJVcsv SHirasfHon 0 Bu 2 S2

Hxo vaDiouccioDolCJal iVV 13 33 DID 1VV 5V ii-L 30J. ivxuuo03015oix ioo 31 OJdeoJV nal-Jsxasxuusv dsvctotlas»usvMJV4>Mds>>3 J>»'J,«l3(q3a.3 SIBTV -562-Irlfs- • 9.Hxo vaDiouccioDolCJal iVV 13 33 DID 1VV 5V ii-L 30J. Ivxuuo03015oix ioo 31 OJdeoJV nal-Jsxasxuusv dsvctotlas »usvMJV4 > Mds > > 3 J >»-3 (I-V3.

VDUUS 3L.V MJVVDUUS 3L.V MJV

03 S3 S1H60JV louiolali 3VVUUU1VV 3V1 sovvv333<c3ovovivsi VVV so<c1313 03V DVV 3U is 11V 3V1 (assxu«£ CSV BI«SCSV J>li>>To«>>-Jot-d 33 «salua.Ji s>«_jdji >3 SI dsvboJV sx_Ja£ dsv auj>\l -§-2cn--- O <1 c u a. a L. *C CJ H C H ca <£ <c <x (/} < X -J « a； <c η H ㈣这 t- o JZ u H <C >.〇 —< o a t3 〇ί H —«X C f-tn «c <« <x u o 1/ z 、i-r x -i H 6- 3 <S c ^ u CJ • X 0 H f- 3 O ai t--J u L- O Oi O </l c o H —卜 ·-» <x a a u u jr o H <E CJ rt H > a 3 <X a> h -J o to o t- o <£ <£ Ο H u a a. u S- CJ 01 o (/) <s 3 <C <C o o M o <s X u 3 sr： c u a u Q.H (0 < <s a c- H «c a t- < \ cxo > ff) < 、《 c: c < o u ir n- 〇 c. o e v〇 Oi o ro to 卜 L· < 01 CJ ¢0 t- ^ a flj卜 > 〇 t- t— V o c〇 <x 0.0 c a Η H V) U >%〇 U H <n o >^o ο ί D <S ilt ο H L 〇 C- U t- t— X Η ί- U H· CJ o tn e Ο O t. CJ a. a -H 〇 <s a 〇 >>〇 o e- U e ai cj c〇卜 <Q <X —CJ < o a u a >> 叫C3 CJ O 3 *X —< < o o .ΙΠ8Γ vv33sv3UDiovvv3Dll:U.LVV VVii UDV 301 «vvuDl iul OVV vl3t-J.Voive-vvlul llousvvuilvovvouo JVM sxaSAn MxoJv«0w>.-1ΐΑΙΛVII uvu UMVL.3CO33-1CG 35a;TI S>,_1oJd •06rn- 1νν11ν131ο03«£ννννννονοο353ν<χ3{-υν311ον33<ενϋοο315αινυ1ονν3ν38ϋ1ον1νν5υ3νο31υννν1ν13Ε-3ο5 vvvvxvvvvoutuuou仁 3.ux3uv3Ivviv.Lvl3vi.L3.L31Qllvliivl.Lo51Q5vvllo3vlI313vll3ixva33118CJ50ovivv3 31vlaolxv.Lula.LLLLi31XD313u3oouuulvlou.u8uvlo3vlul3oilo3oi-ovvl.LVaoulvvvsc)5vvsv3<c3vvov3v3v vov8vvuvvc)i31<s.LLVVl.LoloV.LLV.LUVVOVSDi:}vovvvlvvvvvvvovvvJ-L.LLLLU.VVV33vvov5vo5551ovvlul.LL3vltJ3v 3J-LL3vlu.uvxluyc)VS.LLuvvvvvvvu53(-aLVoualDVVVV.LLu.LLLolullulc3xll3vvoo.ut-u333.uovl3vioSJ8avvv3 ,LLLLLVVVVVV.LL31uvo.L3.u3U.LLyo5voVV.LVovvv5vv3VV.L3.LL81VDi5<_)vs535u515t-51vvlv3(-5isivsvoiv sgl i3av3avvlyulou5ivl31vlxolvo5vvvsiooov3vvvlsvvLLSU131ovv5533volvivllv.uo51uvvyc_)vvvvoi5uo ?6ι (ν>>ΙΟα.ον31ον31ϋ3νν5νννν1ννχ1υχ<-)νο<£νννν1:}ννννδ3130ϊ«ϊ1ρα3103 S3 S1H60JV louiolali 3VVUUU1VV 3V1 sovvv333 < c3ovovivsi VVV so < c1313 03V DVV 3U is 11V 3V1 (assxu «£ CSV BI« SCSV J > li > > To «> &s;-Jot-d; 33 «_Jdji > 3 SI dsvboJV sx_Ja £ dsv auj > \ l -§-2cn --- O < 1 cu a. A L. * C CJ HCH ca < £ < c < x (/} < X -J «a; < c η H ㈣这 t- o JZ u H < C > .〇— < oa t3 〇ί H —« XC f-tn «c <« < xuo 1 / z, ir x -i H 6- 3 < S c ^ u CJ • X 0 H f- 3 O ai t--J u L- O Oi O < / lco H — Bu ·-»< xaauu jr o H < E CJ rt H > a 3 < X a > h -J o to o t- o < £ < £ Ο H ua a. u S- CJ 01 o (/) < s 3 < C < C oo M o < s X u 3 sr: cuau QH (0 < < sa c- H «ca t- < \ cxo > ff) < 、 c: c < ou ir n- 〇c. oev〇Oi o ro to BU L · < 01 CJ ¢ 0 t- ^ a flj BU > 〇t- t— V oc〇 < x 0.0 ca Η HV) U >% 〇UH < no > ^ o ο ί D < S ilt ο HL 〇C- U t- t— X Η ί- UH · CJ o tn e Ο O t. CJ a. a -H 〇 < sa 〇 > > 〇o e- U e ai cj c〇卜 < Q < X -CJ < oaua > > called C3 CJ O 3 * X - < < oo .ΙΠ8Γ vv33sv3UDiovvv3Dll: U.LVV VVii UDV 301 «vvuDl iul OVV vl3t-J.Voive-vvlul llousvvuilvovvouo JVM sxaSAn MxoJv« 0w > .-1ΐΑΙΛVII uvu UMVL.3CO33-1CG 35a; TI S >, _ 1oJd • 06rn- 1νν11ν131ο03 «£ ννννννονοο353ν < χ3 {-υν311ον33 < ενϋοο315αινυ1ονν3ν38ϋ1ον1νν5υ3νο31υννν1ν13Ε-3ο5 vvvvxvvvvoutuuou kernel 3.ux3uv3Ivviv.Lvl3vi.L3.L31Qllvliivl.Lo51Q5vvllo3vlI313vll3ixva33118CJ50ovivv3 31vlaolxv.Lula.LLLLi31XD313u3oouuulvlou .u8uvlo3vlul3oilo3oi-ovvl.LVaoulvvvsc) 5vvsv3 < c3vvov3v3v vov8vvuvvc) i31 < s.LLVVl.LoloV.LLV.LUVVOVSDi:} vovvvlvvvvvvvovvvJ-L.LLLLU.VVV33vvov5vo5551ovvlul.LL3vltJ3v 3J-LL3vlu.uvxluyc) VS.LLuvvvvvvvu53 (-aLVoualDVVVV.LLu.LLLolullulc3xll3vvoo .ut-u333.uovl3vioSJ8avvv3, LLLLLVVVVVV.LL31uvo.L3.u3U.LLyo5voVV.LVovvv5vv3VV.L3.LL81VDi5 < _) vs535u 515t-51vvlv3 (-5isivsvoiv sgl i3av3avvlyulou5ivl31vlxolvo5vvvsiooov3vvvlsvvLLSU131ovv5533volvivllv.uo51uvvyc_) vvvvoi5uo 6ι (ν > > ΙΟα.ον31ον31ϋ3νν5νννν1ννχ1υχ <-) νο < £ νννν1:?} Ννννδ3130ϊ «ϊ1ρα31

Claims

This i amendment supplement 81. 9. 2 6 This month A7 B7 C7 D7 β Six 'application special issue model ffl Annex I (A): Patent application No. 80105045 Chinese application patent range amendments September 81 revision 1. A pharmaceutical composition for the treatment of high blood pressure, which contains TGF-Zhao 1 〇 (please read the notes on the back of this page first and then return to this page) Printed by the β-Consumer Cooperative Society of the Ministry of Economic Affairs The size of this paper uses the Chinese B standard (C «S) T4 specification (210X297 mm)