eesCe5i=:SgpEeSs
`i.ag
`
` th:fee~~fEbOZvegeh2As
`'~Me
`>
`4slew
`
` ¢$22xze2858a58
`
` usQ=ConkSsenegsSjOn_
`we
`>
`
`OZAnomoS©©awn
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`=a=8cS3usa36&Qo2-22£28ee rene©}tot££6uweeg35e5W535SSthiI>5©wooO8O928S§gaS <62ZceS222=&oZz ih,penZeOsn— O50&O.;BiXeNCOatERBOda3geqoaeSaPsatazeeZx2c55oa29& $229e542aOiwoOd222
`
`
`oe
`
`i~355ZO5O29=O
`cyQZzaeot
`ah
`recat
`
` Ba2cDaeheace
`id,
`Nt
`Reag!el
`Zz
`Zz
`
`
`
`
`
`
`=Nisoasé 5”S'O~feiiolTtegySSaf©&SSFey aSxOStiwoiearIroSanata4ZO5502
`
`
`Hso0on
`c2Ge
`
`:
`20543oi+Ls}ctoo
`
`o==S&Sac2©2O
`<2888
`BZG2&ea342S2ue>m0RS2ioa9OFNSgsFiao
`
`tOwWsabataSSOLayOs
`SZSit=@ou&&-<L
`
`
`
` 2Lost<gyoz=S=C«SaGa83wBe.SIi0.83to5eyeSazaoS
`
`
`=>:iaOe
`rha
`
`Pm!123a
`Lu*.=
`ne
`anee=<>SMS
`omZanWw
`ef
`pomod
`
`
`
`—tUT8
`
`pon,£3,
`
`ns
`
`
`
`

`

`(12) HRS AAERA A2 Fh BY Bl Bn IB
`
`(19) thFR4a ik F* 4X SHR
`A rR &
`
`(43) BIBRA0
`2007 “£1 A 25 H (25.01.2007)
`
`
`
`eS) MITN00 01AA
`
`(51) Bs SAS:
`CO7K 14/575 (2006.01)
`C12N 15/16 (2006.01)
`A61K 38/22 (2006.01)
`
`A61P 17/02 (2006.01)
`A61P 9/00 (2006.01)
`A61P 27/00 (2006.01)
`
`PCT/CN2006/001679
`
`
`
`
`
`
`2006 4F7 A 14 1] (14.07.2006)
`
`
`
`mye
`
`HH xe
`
`(21) Bi Pigs:
`
`(22) Elbe dia G:
`
`(25) Hig:
`
`(26) AAS:
`
`(30) fR4E4:
`2005 10083894.6
`
`200510103293.7
`
`
`2005 42.7 1S E (15.07.2005)
`
`CN
`
`2005 4£9 A23 EG (23.09.2005)
`
`CN
`
`
`
`
`
`AMRAAEZAl(BETINGNORTHAND
`(TW) HBA (RAB SP TE es IER eS
`
`
`
`
`
`
`
`
`
`
`
`
`BIOTECH CO., LTD.) [CN/CN]; + HALE THRE
`
`
`
`
`
`
`
`“ESL
`IESE BRS 2ht AE HB 2h Lal A402
`, Beijing
`
`
`100085 (CN).
`
`(72) KAA; &
`
`
`
`(75) RAAHBA (2H £O
`): ## WONIE,Liya)
`
`
`
`
`
`
`[CN/CN}; "FEB AE Be TT ee EK _L HP RSS eT
`
`
`
`
`
`
`
`HE
`
`
`Ea Bs 24 ful A402 8 , Beijing 100085 (CN). GAk
`
`
`
`
`
`
`
`ws
`MA, Suyong) [CN/CN]; Fl dE ae Th eve KE He
`
`
`
`FRRRS S PSN EW EA A402 8, Beijing 100085
`
`
`
`
`
`—|o>
`
`CN). VFR LCXU, Songshan) [CN/CN]; + El db ae
`
` EES S PRN EWE A A402
`
`
`
`
`ye hE RL Mh
`
`33, Beijing 100085 (CN).
`3c 3 E (WEN, Meiyu)
`
`
`
`
`
`
`
`
`
`
`(10) EixasAS
`WO 2007/009359 Al
`
`
`
`
`
`
`
`
`
`
`
`[CN/CN]; 1 BAG Ta EK EMT eS Be
`
`
`
`
`HAW fil A402 2, Beijing 100085 (CN).
`
`
`
`(74) KHA: PRE RRAZRHRBASS Al Bes
`
`
`4 Ait (CCPIT PATENT AND TRADEMARK LAW
`
`
`
`
`
`
`
`
`OFFICE);El dba 1kh K 2-5 0 3 Br
`FL IM@8EE, Beijing 100037 (CN).
`
`(81) HEH RAAB, BREATH DE
`tka): AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG,
`BR, BW, BY, BZ, CA, CH, CN, CO, CR, CU, CZ, DE,
`
`DK, DM, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH,
`GM, HN, HR, HU,ID, IL,IN,IS, JP, KE, KG, KM, KN,
`KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LV, LY, MA,
`MD, MG,MK, MN, MW, MX, MZ, NA, NG, NI, NO,
`NZ, OM,PG, PH,PL, PT, RO, RS, RU, SC, SD, SE, SG,
`SK, SL, SM, SY, TJ, TM, TN, TR, TT, TZ, UA, UG, US,
`UZ, VC, VN, ZA, ZM, ZW.
`
`
`
`(84) REM (RAR, BRA—HTRAEMWE
`+4") : ARIPO (BW, GH, GM, KE, LS, MW, MZ, NA,
`
`SD, SL, SZ, TZ, UG, ZM, ZW), WIE (AM, AZ, BY,
`KG, KZ, MD, RU, ‘IJ, IM), ERY] (AL, BE, BG, CH,
`CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HU,IE,IS,
`IT, LT, LU, LV, MC,NL,PL, PT, RO, SE, SI, SK, TR),
`OAPI(BE,BJ, CE, CG, CI, CM, GA, GN, GQ. GW, ML,
`MR, NE, SN, TD, TG).
`
`
`
`
`
`A Bp A Ai:
`eae Be RARE.
`4hhe oy W132LE ALA BA oN
`A a AA EL TR BE
`
`RMAF
`
`PI AREAKRBREREBEAS, FASTRAK
`SAPCTAGR ANT ede SRA SASRH.
`
`E THEREOF
`
`
`
`
`
`WoO2007/009359A.[IMINOANIMATTTUITIGAMMYNA
`
`
`(54) Title: THYMOSIN BETA 4 DERIVATIVES AND US]
`
`
`
`
`
`
`
`
`(54) RAAB PR: AAR B 4 ATER
`
` YF
`
`(57) Abstract: Novel thymosin beta 4 (TB4) derivatives and pharmaceutical compositions containing themare provided.
`Suchderivatives are Gly-TB4 and Ala-TB4. Use of these derivatives as pharmaccuticals in the treatment of skin lesions, heart
`injury, cornea injury and coronary heart diseases are also provided.
`
`(57) iz:
`
`PEGE T or WERE B4 (TI) HEM RSAEMAMAS. ERM Gly-T64 A Ala-Té4.
`
`th
`
`BERTERTTTTRRRTAG. LEAR FRAGF/RESHNFI
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`ROARS B 4 HTReFyA
`
`FRARAR,
`KARAARAM IRE B4 (1B 4) AAGly-T B 4 Fe Ala-T
`64. ARARUASA MIRTB4 HEM MSY, KAA
`BPR TB 4 TEA Fae RRR. SARAER AAG
`ARG Fe /RDS MAN BMA. ARVIEPRAA PR TB 4
`AT IG FF ROARSEER TAA OS ARCB OR TA. RAFe/RFESY
`FIR tk.
`|
`
`RAR
`RERBRAARMAGHRA. FSARARS HY EZATK
`2H). Bak Cealaze Bake, MO ARATRE ORA. AME
`PLES ES. BRL RAS (AME) ATH IRAA
`EMBRACE, HPT BAI LAA FeMHA
`ESARRAS. ARMA, TH42ColdsteinFA,
`(THE
`JOURNAL OF BIOLOGICAL CHEMISTRY, Vol. 257, No.
`2, pp. 1000
`-1006, 1982, Chemical Characterization of Thymosin beta) m
`FAD AIR RRA AREALS (TFS) PAB wy RA —APS AK,
`B43RRM, woSEQ ID NO: LAT, PFE4.96IKD, AMS
`HEBM.
`TB-4 RAMEE SERATRANHEA, SHES
`WE ey HO WY 3 AG FEY 7m 2K HE om J LAE ( Thymosin beta 4
`activates integrin-linked kinase and promotes cardiac cell
`
`migration, survival and cardiac repair. Nature, Vol. 432, 2004
`#11A 258), LARRARRN, RHEE RBALHEHHS
`RETBNESe IE BARAY UT FH IR) ERO TG MALE. TB -4
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`AJ FeRRS, VERBAG HER. FHAR HE BAA
`RK, TR-4 RASA EMF 1 RREHR.
`EteEGR TB 4 AQISPEPTRAYEE TAA 4 RA,TB 4 HY
`38 bnopt 3%Uy th PAAh ASUSIEE HEA, EK RERH
`AEIUGBURA EEE, AROSEAFee Akt Ae
`WHR, EA RM, PANTSRARARARAR—TA HG
`FF AAR.
`HE RPMNAPMARARALALRY ZARA-PHE SER
`FitsMEO (FP:TB-4) BSRRAEGP SIE
`Bra HAZ, LLAAREEYRAWRARAA TRAIN
`PBF RAHLT RAR EPAR A SERMAREAA.
`KAWAIa Ce TR-4 TRA RERAMAM, RAB
`ALF IMMRE A NARA, HABA RGA
`ARMAAER SRARRUELSA OEEALATARIB-4
`Mat TR -4 tte,
`
`RARE
`KRRAW-ADH, PRA1B 4ATA, Hoa A SEQ ID
`NO:6 Praségy Gly-T 64,
`42SEQ ID NO:5 ATHY Ala-TB 4.
`KARIMRAHH, ATH Cly-TB4 WARAla-TB 4VA
`MitRASMARMFERSUFSRMAKHS. LKRGF,
`Prk Cly-TB4 ARAla-TB4 SMUKRALABSMRANAERG
`a4 «
`
`ARRMM-ARBARY, WitteFHRRG N- Kiar
`aMARK B4 ATA: Cly-TB4 KR Ala-TB4, GatoFR:
`1) RRA AR
`eit 240TB 4ARES (SEQ ID NO: 1), EA AMATSF
`
`2.
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`FURYAARBTTyER
`$FSAAR TB 4DNA ARS,
`HlZN R3eAe C RIGALIRA, EAT N- RSH, Ft
`3) 4, TB 4 ATA: Gly-TB 4% Ala-TB 4 ABAD. ROA
`GST RABFP NIA KOEASRETSRASRATE DNA
`5 LAASTPTRRBN RGAE C KHAaLR
`tt, FRAT URBRF A.
`BPA KwLRASOAFKP, 184 TEMES VAR Sip 3
`JApBSK AIRAEA eR, BURSAERRER-RER
`WAAT4 EIR A, Ba, RH EAT.
`Aa iT ROSS RA BHAA S A RaBH
`a ADApOM (AMRRAFMARRAMI, BARBRA
`CSTERAEA, AAADHR) HH, AMBAANEAMEAT.
`2) STR 4TEMNRABDE BME
`MeoLikPPPSSHERAGER, ARM ARAR
`Ria, AGEZAM, BAHEO SARAH LEA. BEAM
`pure BHR SSA MHALAR,BLAALAA OST
`Hoth, ABLAMHASZOHAROBBWS, Beea
`BtBUT, RERAEGMERKT UWNHAPM.
`KERMA —-FH, PA-HGMUSCH, KOAARZA MIEN
`TB 4 4744: Gly-T 6 4 Fe/& Ala-T B 4,
`VA BAIAYY AkTHES
`8 BM
`ERA He, RE BWLTRLH’” BRARARRAAN
`PATH, SNE ATAM. KiB “RUA”A.
`7] (Blte (RERMABBE) PKA) AGA. RAT AMARA
`18 FFF) BOA
`kB ARATUARABAA, iekieid, LIRA,
`Shin, Hn, RAMI, BeebhAwh. ABI. Tih. SARA,
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`Bwewe, BRABABABSON, KARAM RA. BKER
`UL BIKE eaFoA ERA BAK, HAA TE
`wim, SRARABI. HAA. F488. Re. WR.
`ZR. ARK. GP. OS. BR. RRBA. BRR. A,
`RUL4GH. WP. the. A. OBR. RK. GH. Beeb. WR
`EB, BSWTUALSRY KEY BBR FUGA ois BAA,
`RE pHBHPF), MRMASWAMARRBIR, BHR. FRA, HF),
`FUP. ARR. RAHA. BAA BA,
`ERRAND EMARP, PRAMAS ART ERA
`X, HSA 0. 01%-0. 0H MRAACly-T B 4 & Ala-T B 4,
`VAR SUES, ARAWETESM RA,
`BRR Spite]MEREBAS. 4H
`FB) Kl LIGAAIRS Bob, ideaRRAL RAL RF. OAR,
`RA (BRA). BRK (HseHB). BAR, FeAeA. A
`REMKREARP, RR PARES ith] RIESBIAL K
`F. LAA. CE. RAL RARE TFARMAAMSH, BF,
`ATBRRKA BA HASMRARSBEKERHR THER, WRE
`BZ, ASMETV ASBA FeEyEAhe KARA AEM
`
`KRRAN-PRBEARY, HARMSOWARRA, HPSH
`50-500ug/ml 44RRAGly-T B 4 #2/K Ala-TB4, ARHA
`KRERRAAYBHREAE, VBBZMLTRSHRA ZARV
`WA-RMFARP, HRASMARRR, HSA 50-500ug/ml
`84 AA BRATEW Cly-T B 4 Fa/%& Ala-TB 4, BHHAAMRA,
`RAAELTRLMRA, ZARAHA-KRARP, PRASW
`ABR, HPSA 50-500pe/ml HMREM Cly-T B 4 Fe/H
`Ala-TB 4,2SOW RRA, RSRRAAVEZ, VBZOLTE
`-4.
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`ZO RK.
`HERRERA MASH, NTShe RAKE,
`SUR. RRAME. Pea). BR. HA. HRA. TRA. BR. GL
`ROBAREAARERARAMAPMAEBA. SM GlHO CF
`BF i i, 25AL fa FlHE) ( Remington’ s Pharmaceutical
`Sciences and Introduction to Pharmaceutical Dosage Forms),
`$197k, Mack BRAS), HMM, BHREL, 1995 +.
`MEARTREARNSUA, BHRALSS ARAM MAH
`RAR —FP RS AHR A LAARAAT RH RHHES
`LEAR HAGA, SiHHF LISAARTBR. BHR. UR
`me. GLH. AE. BH. HH. BR. BeeR, WREZ, KA
`BORSA (HieRRA. BEA, BRAMTA. RB)
`RAMUS, Biot] wREE, KE SEH HBA SA
`aieTELEEASA, RPRAKS OHARA ERAN EE
`RFOEZESAMEERY (BRERA, Bie RF) HS
`MALBRMP. WKREL, BTUAAMASwPAMSVAP Bm
`WAR MLIA, LEMRT PAARLBe.
`EKRRWMHR-RHARY, PRAWASMARKEBA, FH
`ASAT 0. 01%-0. 204gARKATE Gly-TB 4Ala-TB4, Fem
`LyFES 04 IA Fi] eeLEH,
`ERAWMAK LEASH RAM, NOM Bel mR
`RSA REN, BEA MANHBA. LAME, RRARH
`SF AHRDSBI PRATORSIEAREAL (GridHLSAL
`SA2E PR. —MOMOEM. —AtK. AFCSBEE)
`LARA REEMMBA HEARNERRAARHRE. £
`WEREA GML, PURTRAANKRARA SHE, WR
`HERE, RABRKASPEAHRAMRER A (olde ey
`
`~5-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`BR) LVR RSAMSWEEDREAdeSUREDDAR
`AD
`ERRAQL-RHARP, HFRBEDMSCMARA, HPS
`A 5-500ug/m1l MARATAM Cly-T B 4 & Ala-TB 4, AAO. 003%
`HAE OR, FBHMLTRLHR.
`BRAVA AKG IRAGA, MASA Beel A A. BR
`©. a. RRE. BR. ASR. BaehAKHWA. TUMNF
`RFRA BATELRBAGHSA (PieMRRERED. RK
`CMe. RAAARPRABR ), BR (seGlHe. fhin A
`H. ABBAS); HH (PWR. HG. RAED), DHF
`It (PeDASRED CHB );, STARA ( O]ae A AR
`4h) HASURR. PRAMASAM AEARA A. AF
`FREE RAS TARR RR, ER, REF RHY
`XK, ARH ENTOAUF RFE BHBABA, RAWARRAS
`RIE.TVAFALFRARA TEL HAH, He RFA (4
`jo RBIR. FRAME. RAMEEAA); FEF] (Hl de HP
`FENG ATABTA); SEAR( Pl dedead. eR. GR.
`SyaA); VA BGBF ( Hide Ff RRA B-M-RAR PF RB
`KLE) HSERANA. HARTA ASCHHH. FHA.
`HERA. FoeA. TATE HAC TOaARA
`LBB, ARFRRAMGEAREF LAR.
`RRA WAKER A FMR ( PwTPERE
`He) MAA HASH. AT ERHERA TAU SAR
`GBA OPH SBA (PlwHRAPRESHEASS ) FE.
`Bay VA REhoREIN PO IFRR RGR
`EBX, METAS RHF, te BFA. RSA. Fe/RAK.
`RA, SAMATARPABA, ARAN SEINE (Fle AB.
`6-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`ARARK) BH.
`TATRAAM BSASEA FARR
`ARBEEAMRARH. EREKKEARY, BTMWIMEVA
`ayANE Lde42IREEAAl 7K USP; REAM LAIRT ROE
`Sty. Ringer KZA. BAER. HAEUCR.
`VARSLBA Ringer KiEAIR; KARINE LIAR CH. RO
`8. FPRARB;, UARAEREIE 46/48RRS BRI. HI.
`i, Sib. MOR. ALBRHAN. PAPRATE.
`KREWMMR—-AH, PARA Cly-TH4 FAla-TH4ZTBH4 N
`RNSTA RGIS SERA RURRAARARA. FAR
`TG Fo /SES AY BHA.
`KRRAABASHMAM, AMARAKBARLASIE
`speieR. KARR. ARRAN EEA, RAW
`4 Gly-T B44 RA ALATARIBD A HAE.
`KEWAGABET Cly-T B 4 Ae Ala-T B ARP SMALE
`A, RARE 60 RRAK RW ERARAIRWSAR, MK
`HRAKREROARENEAEILARARS., ERERMRET
`Ady Gly-TB 4, Ala-TB 4 RMRETB4 AMARA SEF,
`sak Tete, HASERABKARET SHED, MH Cly-T
`B 4X Ala-TB 4 HG 77 KRTR4 WB ARWARMAMBTE
`Yy Gly-T B 4 Bu Ala-T B 4 34) GECR Smaaese we RRA
`Ss MLSHI, 22e beLARAS BARe REAL.
`ARAMAT Cly-TB4 fo Ala-TB 4 PLMRRRAR AG
`Al, RAR 20 RRAKR MORAG, AABAIARRRM
`FRE), HEGREGly-TB 4. Ala-TB4 & TB4 S RMKAY
`He 0 ee eTRRs 50%, to Gly-T B 4 & Ala-T B 4 Wie yy BOR
`TB 4 WAS.
`
`-7-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`KARAT Cly-TB4 SHARE, RAR
`130 RRAKB HY AR IN RUG, Atea] AR A aR
`Al, AS IRSAWAREK Cly-T64 R184 4 AWARWRMARE
`AWE, MALBARMAHHMARE BAKE, CRSP Sn
`RFE Fe FM,
`LRA RRA, RAH TB 4TE Gly-T B 4Fe/HAla-T B 4
`AGF SURG. RRARREURARARGGS a HA
`AB ATARTIB 4 Oe,
`KRG YWL—-FA BBA ARTB 4 We Cly-TB4 Fe/K
`Ala-T B 4 i697 SAM BAR ZAM SIARUAG. RRBRUAGM
`RABBRRAAMAK. ARRAN -PRMEARYE, MRS
`RELA. RRA RARVA A REAROH LISAIRS
`AM RIA ARMEMRAA IE TR 4 AM Cly-TB4 Fe/X
`
`Ala-T B 4,
`KARRBBRRAR OR, BAYMAK. WRAA SHAE
`Fi YATE. FAVAAW AR OH. HR, PALER (6)
`WRAL MAL PRAR KT). Bee (PloRIGA) Ri Aes,
`Fl (we IRA) SARA. WTR RAPA AA AES
`Be AISAMHEAAR. SHNANSA HHA LRH AWBSH
`et BA ROG SMEAR. RRR ARR
`“a=.
`Rit, MFOHARW AIRTB AEM Cly-TB4 Fo/K
`Ala-T 84 WHEwWBOwW, FETFHR-AHNEPHEKAA
`100ug-Smg. SE ASH So VHARAMEYRAAA AMF, (2
`—FRAAKH 0. 1m1-Sm1.
`
`LA
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`Bl St ARMA ERTB 4Fe Cly-TB4 BM ARSEERR
`6) 33 9 28 RK
`Ad SxtA RPA EA TB4FeCly-TB4 EMH ARSEEYA
`AGEFLW BS 28 RR
`B32KRU TB 4 fe Cly-TB4BH XKRARGUR
`PFLLVS R.
`42a K RAAT B4 AeCly-T64 GH AROMAS AR
`RBI BALL BS ER,
`BS 2x KAMA EM TB 4 fe Ala-TB4 BM ARCRERR
`84 33° 2 OR
`A 6 txt A SABER TB 4FeAla-TB4GMAR SRSA
`ACFEFL 09 33 7 ER
`A7 2st XK BBATB 4feAla-TB4 EH KRARKRGUR
`4 it TA] 9 Bo eR
`B&8 Sx KBR EA TB 4AeAla-TB4 EH AKRARGOR
`JRAZJL 0 BS 9 ER
`Ao EHRRARPG TB 448 (C). Ala -TB4
`21(B) fe PBS 48 (A) BLAM, MRA ARELRRRBRMAR.
`B10 ET RA A Rae ARe! $A TB428 (B), Ala -TSB
`428 (C) fePBS48 (A) BRSAN, MRA ARELRKRAHER.
`
`ARAB FS FALE A IAAT,RAAF 2006-7 A 5 8
`FAIA LALSAR S48 wRPGMT «6 4-A/BL21 4ePGMT
`B 4-G/BL21 RTP AMARR LARSHERED PS
`(CGMCC, aby, PKATAL—#, )
`, HRIR-FH CGMCC 1750 Ae CGMCC
`
`1751.
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`UAE BEA RY fo KaeRtAAEEA (2 RT
`RB AA EAERA
`
`KA1. MARE B 4 Ay Cly-T B 4 wy ls
`1. EAAAS pOMT B 4-6 Wy
`AR4E B4e TB 4 BAB F (SEQ ID NO: 1), deF:
`
`Ser Asp Lys Pro Asp Met Ala Glu Ile Glu Lys Phe Asp Lys Ser Lys Leu Lys Lys
`
`Thr Glu Thr Gln Glu Lys Asn Pro Leu Pro Ser Lys Glu Thr Ile Glu Gln Glu Lys
`
`Gln Ala Gly Glu Ser
`
`(SEQ ID NO:1);
`iFE] KMATEFBTSR ASAT B 4 DNAS PHY Gly-T
`64 BRAD), AFRABEMRH, ANAM MT HRS AZ N
`AS Fa C R34AoLRH HBSS BamHI. Xhol, A N R3S14 E+
`KAM Gly, ke SEQ ID NO: 2 ATA.
`
`GGATCC GAC AAA CCC GAT ATG GCT GAG ATC GAG AAA TTC GAT AAG TCG
`
`BamHI
`
`AAA CTG AAG AAG ACA GAG ACG CAA GAG AAA AAT CCA CTG CCT TCC AAA GAA ACG ATT
`
`GAA CAG GAG AAG CAA GCA GGC GAA TCG TAA CTCGAG
`XhoI (SEQ ID NO: 2 );
`
`SOP RRaBeFRRA
`FL PFTBL 0RB 5 de F
`
`Gly Ser Asp Lys Pro Asp Met Ala Glu Ile Glu Lys Phe Asp Lys Ser Lys Leu Lys
`
`Lys Thr Glu Thr Gln Glu Lys Asn Pro Leu Pro Ser Lys Glu Thr Ile Glu Gln Glu
`
`Lys Gln Ala Gly Glu Ser
`
`(SEQ ID NO: 6);
`ARG Bde CST BARR TIAMAT det BETSRASmR
`KF AK ODNA HF) LEN, C RGGA0_E PRBes EcoRI. BamHI,
`~10-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`43-3) AWAKE AP|, te SEQ ID NO: 3 Fae:
`
`GAATTCATGTCCCCTATACTAGGTTATTGGAAAATTAAGGGCCTTGTGCAACCCA
`
`EcoRI
`
`CTCGACTTCTTTTGGAATATCTTGAAGAAAAATATGAAGAGCATTTGTATGAGCGCGATGAAGGTGATAAATG
`
`GCGAAACAAAAAGTTTGAATTGGGTTTGGAGTTTCCCAATCTTCCTTATTATATTGATGGTGATGTTAAATTA
`
`ACACAGTCTATGGCCATCATACGTTATATAGCTGACAAGCACAACATGTTGGGTGGTTGTCCAAAAGAGCGTG
`
`CAGAGATTTCAATGCTTGAAGGAGCGGTTTTGGATATTAGATACGGTGTTTCGAGAATTGCATATAGTAAAGA
`
`CTTTGAAACTCTCAAAGTTGATTTTCTTAGCAAGCTACCTGAAATGCTGAAAATGTTCGAAGATCGTTTATGT
`
`CATAAAACATATTTAAATGGTGATCATGTAACCCATCCTGACTTCATGTTGTATGACGCTCTTGATGTTGTTT
`
`TATACATGGACCCAATGTGCCTGGATGCGTTCCCAAAATTAGTTTGTTTTAAAAAACGTATTGAAGCTATCCC
`
`ACAAATTGATAAGTACTTGAAATCCAGCAAGTATATAGCATGGCCTTTGCAGGGCTGGCAAGCCACGTTTGGT
`
`
`GGTGGCGACCATCCTCCAAAATCGGATCTGGTTCCGCGTGGATCC
`
`BamHI
`
`(SEQ ID NO: 3);
`
`ERAEAA, Bde (PFRE (F—M): RHE
`Hy) (1992, SRBARE), HihikslonAaAHR, Cly-TH4u
`Be fi RH. pBSK (Stratagene 24] , HF 212205) A IRA) MAee
`BcoRI, XhoI, BamHI 43, H+? EcoRI #2 BamHl WAAR, Xhol Fe
`BamHI 49 Gly-T B 4, EcoRI 4e Xhol + iitRH pBSK, Bi RAHS
`RAS, BAR 1: 1A Th EAA, Bae, eh eee
`4yk3& Cly-T B 4 GRP FRAFRBSE ARAM SARE, HH
`pSk- TB 41.
`FPi FAL REA Wet FRBSE AAA, BRS
`&ARA A RAZ Xhol Fe EcoR I MBE JA pGM ( HLA EAR
`EAMAPIL, HKERIA GST KREEA, HAAADNB)
`HEIKKAK RK, UWCAMARMAAY ERA MAA KRHA.
`BilgeFF MRHRSA KA, AB) EAR SAR
`-li-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`#i, O74 4% pGMT B 4-C.
`
`2. SA S20OMT B 4-6 HRA LAA HS
`SRG FA CaCl skeet KATBL, RRASA 50ug/ml FARE
`AY LB-FR, isles, Aa) SAbikS40WA pCMT B 4-6 44
`FELL.
`FE LK ARIEFRR We EHRPIRFELT Z 0D600 % 0. 6-0.8 A, Ae
`IPTG 0. 1mM HH 3-4 pap Boeke BrA,
`9M RERBE 15%
`A.
`SDS-PAGE WK HIL—# 30KD AAHRAB, AiASH 40%. AT
`P4 HPREMALREe, LTMARM. RE SGRMAAMR
`Bit+Gly-TB426HRALEA, ZA PCM2TB4/BL21,
`HRERAMSAAE, PRHARKRET GRMRAMREMAD
`Gly-T684 B4HRALHH, 4% PCMT SB 4-G/BL21 (ARE:
`CGMCC No: 1751).
`
`3. Gly-TB 4 RARBY
`1)RIL: HoLRaSHS Cly-Th4EHRAHRALH
` PGMT B 4-G/BL21, AA 50 ug/ml FRBKY LBIKRYt
`BIS (37C, 200rpm), HaeAARHL 1: 30 HAPBASA 50ys/ul
`FREE LRA P, 37CHR 3 -LHG, AeA 0. 10M IPTC KH
`H 4d AT KRA SDS-PAGE BIRDAT, RILAREAAD Gly
`TB4 GOD UPBRRAHE, RAZ EBARZEAM 60%,
`2) KALARik:
`a. SEFRARAM:
`a) PF RISA (LK):
`Ree: 10 R/F. EH: 5 R/S. BUE4A : 10 H/F,
`EMER: 50KR/EF.
`
`-12-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`b) #P HRA (15 FH):
`BERS—4A: 315 He. PERO AAP: 100%. BULA: 10.05 K,
`R4E: 50. REAR: 90K.
`VALRARE RE; FOMRPPRR EMAAR,
`BRERGE: 15%. BBR: 450K. Ah (500 K/F): BAAR.
`vb AHABRAIL:
`i) PFEFR:
`MOKPOEMAH, HEAD) 50 SF (250 SHHEA
`HR) LK, 36°C. 200 H/F. 8 DRGIR 50 SHH TREES
`700 SAP LK +,
`36 °C. 20048/, HARTA.
`1i) RBAGERAWE:
`HIEFRAT 4 APT IR (0D600=3-4 ) HABE, HIFRAI, 36,
`150 48. 33.100%, FHSAA, SbAIAYA 2. 5 teAe 2AT,
`2AA 800 SHAM, AeA 1 H IPTC HH (ZA).
`3). Bat:
`(i) HAeBAT
`XA GSH-Agrose Bare(Pharmacia 24)), FARA 25mM
`Tris-HCl, pH8, L#+#e, RBOHASLAL Glutathione
`Sepharose B#r4z (Pharmacia 4) ), #@PGAS 10M ZRBSH
`TAK (GSH) HBORIAAFREES.
`(i i) Bee
`L— LRAeei(5 NIAU/mL ) 37°C By 2 4 AT.
`Gi ABT RRA
`KA) Source30Q Bat (Pharmacia Aa) )JVin, FATAA 20mM PB
`( BERRSL—4A — BERR— BUFR), DHT, HERE (2 ) BAREB a
`on Ht] RARE 1ATLA, PBRA 20mM PB, pH7, 0-1M NaCl
`PRAM, KAA HEA ABE,
`-]3-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`4). Rosi
`AitbkACHREA] 4 Cly-T B 4 3hit SDS-PACE 2bMl Fe
`BAG HPLC HM, BEAF IB%,
`
`Kiet] 2 MH B 4 Th Ala-TB 4 Ale
`1, BAARAL pGMT B 4~A 44H]
`tka Bde TB 4 BABS) (SEQ ID NO: 1), ZEA ARATE at
`LFS ASR TB 4 DARI, FB) Ala-TB4 ARABS, AA
`RARER, MNAA TARAS KZN Ria Ae C RSgA0 LR
`a (M4 BamHI, Xhol, 4 N RSS _b ARM Ala, ko SEQ ID
`NO: 4 Prap:
`
`GGA_TCC CCT CGA GCT TCT GAC AAA CCC GAT ATG GCT GAG ATC GAG AAA
`
`BamHI
`
`TTC GAT AAG TCG AAA CTG AAG AAG ACA GAG ACG CAA GAG AAA AAT CCA CTG CCT TCC
`
`AAA GAA ACG ATT GAA CAG GAG AAG CAA GCA GGC GAA TCG TAACTCGAG
`
`XhoI
`
`(SEQ ID NO: 4);
`FL? FRal eeBRFR
`FPARTBL i ARF Dl ho F
`
`Ala Ser Asp Lys Pro Asp Met Ala Glu Ile Glu Lys Phe Asp Lys Ser Lys Leu Lys
`
`Lys Thr Glu Thr Gin Glu Lys Asn Pro Leu Pro Ser Lys Glu Thr Ile Glu Gln Glu
`
`Lys Gln Ala Gly Glu Ser
`
`(SEQ ID NO: 5);
`
`‘koe G40 CST RAR FA AAas BATARBSR
`Fo} AK DNA FF) ELAEABFIRS BRS N KSBFe C RIB A_EARAY
`Mie4xEcoRI, Bam 1,3) 5,4ARF He SEQ ID NO: 3
`-]4-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`i)
`RARERAAK, Bide (PFTRE (F=—M): KHL
`#) (1992, ARBRE ), HebikHlaAR Dl Ala-T
`B 4 ABR FTVA Sip UEJAS pBSK Fl FRA) HEA 4 EcoRI. Xho! . BamHl
`dese, 3L+7 BooRI Fe BamHI WA-FAK, Xhol Fe BamHI 4 Ala-TB 4,
`EcoRI 4 Xhol +9 ist/RHE pBSK, BH MAHBIR, BER:
`1 AOA T4 EAA, RL, eh PS) LSEe Ala-T B 4 RA
`AN FrAPERKBRSE ARAM SLR, oA pSkK- Th 42,
`ARRA it TPE ROR AAR, BERR RA
`#5 &% Xhol Fe BooR | MERWEAHS POM ( AEA EMESREI
`AIL, BAZAROST ABSEA, AAAADAR) HMKKH
`A, 1SCHT4ERBAAP ERA MARTER. Bie
`FM eAAeSF peAA, FAELABAHEUARE, oe
`A pGMT 6 4-A,
`
`2. SAARSAREH AA LER aS
`ORG A CaCl ikeeUKMAT A OBL, ASA 50g /ml HARE
`Ai LBP, Pus AB, BAS LikSAHpOMT B 4-A wy
`BEAL
`tn
`HE LK ARSERRA ERATF AZ 0D600 H 0. 6-0.8 A,
`IPTG 0. 1mM HH 3-4 PH RBoIkBAA,
`3M RERMBS 15%
`A.
`SDS-PAGE BIKA RP—# 30K AAMHEAR, RAZA 40%. BH
`RE P4AHPARRAARPREE, PRMERM. RG RMAIA
`RATED Aa-TB4 BOHRA LEY, 674% PCMTPS/BL21
`eR ERS AR, PARR T GRAAMREMED
`Gly-TB4 HH AALHA, PLA POMT B4-A/BL21 (RMF:
`CGMCC No: 1750).
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`O10 H/F
`
`3. Ala-TB4 RIARY
`1 )4hHLRIA: HieLRBSHS Ala-TR4SHRAHAALE
`#) pGMT B 4-A/BL21, A 50 pg/ml FABRALBISRAY HA
`AEF (37, 200rpm), HH 1:
`30 HA|DA 50 ug/ml $MEK
`4) LBSEFAP,
`37 CHER 3-H, AeA 0. 1mM IPTC HH 4By,
`KeBALE SDS-PACE BAYA, RILSMREMEW Ala- TBA
`GUD UATRRRAHE, ARAFEBAREAH 60%.
`2) KAURRid:
`a. SFRGAM:
`a) FPF RISHRA (LK):
`WEAR:
`10 R/S. BE: 5 R/Sb. BURA:
`EME: 50KR/EF.
`b) #PHRAR (15 HH):
`BeS—4h: 315 3. PERKSFP: 100%. BULA: 10.05 2%.
`RULE: 50 3. WREAK: 90%.
`VALRaAREY RG, FRO RPRUERE SAREE,
`ARURGA: 15 3H. MBE: 450%. Het (500 B/D): HAAR.
`c. RERBHEAL:
`i) RTSER:
`MTB PILERRAA, FPS) 50 SH (250 SHH=AA
`Hi) LK P 36°C. 200 H/PsR, BMSHI 50 SHHTRE
`2) 700 SA LK YP,
`36 C. 2004/, RAR,
`ii) RBAGERANE:
`NESEFRAT09 FeFR (0D600=3-4) MABEL, FIFRAB, 36.
`1504¢. WH0100%, AIAB; SNGARY 2.5 RMA,
`470A 800 SHAH}, AeA 1H IPTC HH ( =] Ht ),
`-16-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`3) Bar:
`(i) FABat
`KA GSH-Agrose Batsyye (Pharmacia Aa), FBPARA 250M
`Tris-HCl, pH8, L# Pe, ROWABskwA+E Glutathione
`Sepharose Kapzz (Pharmacia 24] ), ##G AS 100M RRS
`HARK (GSH) WARKFRCES,
`(ii) See
`Li 4 BLAeine (5 NIHU/mL) 37°C By 2EY,
`Gi) ABTRREM
`KA Source30Q A4#f( Pharmacia 24] Y/R, FBR 200M PB,
`DHT, HB (2) GeeAES aea KARA 1ATLH, PALE
`FFA] 20mM PB, pH7, 0-1MNaCl MHRA, KREMEH ARB,
`4) AI
`4¢-3) 49 Ala-T B 4 383dSDS-PAGE HEMl Fe4H HPLCHA), 2
`ERKF 98%.
`
`Ke 3 MAE B 4ATA Cly-TB4 feAla-T64 SARMRET
`B4 XA BUS AERuoRA 09 18 IF ORI
`
`1. FARM MRe ORES RARAK
`AT ipFata Cly-T B45 TB 4 By SRR oRSM
`Ai eee, ZL SASHA Y BAD MHA—KB
`FERRARA FRE
`MLA Eat Ff AR Smg/ke; SVRBA 5SOmg/ke AB, ABR. a
`HHBARWMME, ZSTREAABFARG, BEFKG, #
`REABRATHETER. WFKRWHMEREK, WHF 3 BN 6
`fay, sb RE, HAIR —MN, TACK, Bh sik, MERA
`-17-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`FA) 6-0 Baw4LH44S NELATON #4 3Le2
`PRA ATEX224 2) 3mm Sh,
`€, LDNEHH, HUDALHE, HOMHMPSMPRS, FB
`SESBK. FREMA ERRASGER 3mg/ke/K,H3KR, FiLRH,
`
`2. AARP SHRMLRAS SHEBE
`1) Gly-TB457TB4 ey reae
`RR 1) @LARSMADRAS, PARE Re (He
`2820 R, ERABK 200u1/R/K), FabextRega (#40 20 R,
`BIZARARE TB4 10ug/R/K), Koren (42 20 2,
`REEAREAAGly-T B4 10ug/R/AK) BH.
`YeBh GF 14.28.56 KAAs PEASBHy A( Live 3D Echo 7500)
`RAT (probe 15-16L),WMESMAWASENBBE, Ri Ff:
`Ra(FH OA, SHAT), PER Rea 0.26824 0. 01, Fabeat
`PRGA 0. 2513 £0. 032, Gly-TB 4 4a 0.24994 0.0225, ST HMAB
`SuAH. BH 14 A, Wes eee 0.136540. 0125, Kabeatpeda
`0.1919 + 0. 012, 5 ex} RR 4a4a rh, ASSFH(p=0. 048<0. 05),
`Gly-TB4 28 0.21624 0.0078, SM Mx Reng, ARSwREZH
`(p=0. 003<0. 05), S Raperpgeazaren, AH ( p=0. 086<0. 1),
`IKMLAE RBA, Cly-T B45 1B 4 aby RRBesMeRea dey
`ASSBER Wey (ILA 1). RP, FR Cly-TB4 sleaze
`ZWBERSmy RRRARAMAART IB 4 aR.
`
`2) Ala-TB457TB4 a bak
`RPR 1) SLKASERGRA SE, Palastwee (Be
`G20 RK,EBB K 200u1/R/A), Pobext pese (Aen 20 R,
`BERARBRE TB4 10ug/R/A), Reese (44 202,
`iEAREAAM Ala-TB4 10pg/R/K) BH,
`
`-18-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`9BJ FH 14.28.56 RAYAMEF3) A( Live 3D Echo 7500)
`RAT (probe 15-16L), MRAAAWAS EHR, ARio hk:
`Bae (HOR, ABA), Weert Aase 0.25944 0.0225,
`Fa
`EXT RR 4H 0.23784 0.018, Ala-TB 4 4a 0.243240.01, S#ZMA
`BEwZFH,
`23h 14 AMER RRS 0.130640. 0128, PAvERTRa4A 0.1215
`+0. 013, Ala-TB 442 0.129940.011, 44oz MARSREF,
`23h FH 28 A, MR AR eH 0.120740. 0151, Fapestpa4a 0.1233
`+0. 0071, Ala-TB 4 #8 0.128840.012, AeazMABSHEF.,
`2H 56 AR, WR eee 0.126540.0125,
`Fabext Reon 0. 1232
`£0.0057, Ala-T 64 28 0.167240. 0078, Ala-T 64 ZansfkAus ¥
`BYAEB34ae ( p<0. 05).
`KERB, Ala-Th4 RHRMAAOMRE I RMYHAVES
`BR BY (SUAS). EP, AR Ala-TB4 MORAC EES
`BISRR HABAYAK F TB 4 HR,
`
`3, FF aor BA peas Bf oReae
`RPR 1) @ZKRVERORAS, PRAWNSBA (
`ZA 20K, EAAERK 200u1/R/A), Peepeese (4H 208,
`BUREIER/ARA TB4 10ug/R/A), Biese (42 20 R, BRE
`ATURE Cly-TB4 1l0ug/R/K) B35, 2 RieFe:
`EBBG FH 56K, Rik, USARMAAL, Ry se, HK
`AWA. FA ne REMI 24 ba, HSK ARH, UE
`AT Masson’
`trichrom#é, $4021] LARA T Image—pro® PLUS
`ver. 4.1 (Media Cybernetics) FH#,
`1) Gly-TB45 TB 4 eorex
`SMA ARRAS ESAS FSMeA, Weert eee 30. 02 +
`~19-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`Paves Raga 23.1542.9%( P<0.05 ), Gly-TB4 28 15. 36
`5.1%,
`+2.7%( P<0. 0005).
`HLAABRER, Mex Rees fapest esa. Cly-TB4 2c ihl
`ABSUARH, HHGly-164 MAARRSRAH. HRSRAA
`Gly-TB 4 fe TB 4 3467 Bum8sUR | eg LF HER (LA
`2). HP, PIR Gly-T B 4 F) ACPLAF EL BY RABAT
`WMAAMBT 1B 4 HRA.
`
`2) Ala-TB45TB4 4 reaR
`SAL RRS SAS ESM RA, VR 4a 28.134
`2.1%,
`FaPERPPR ZH 22.9142. 3% (P<0.05), Ala-TB 4 28 16, 862
`1.8% (P< 0.0005); Bex pa Za fax pasa. Ala-TB 4 ozAy
`BHUAH, AR Ala-Th4 MHAERLSRAH, RRARRA
`Ala-TB 4 Fa TB 4 34687a SLR§| A SHLF EG (ILA
`6). SL+, APIA Ala-T B 4 3] ARPANS LA AL RY RRABAT
`MAA MT 1B 4 mR.
`
`4G] 4 Gly-TB 4. Ala-TBP4 5 TR4AMKRRRAM GRA YF
`
`1. MRAMHAE SARA
`AT HAARGlyTRASTR4AGRRRR MEE,
`BET RRM HARDEE: KRARMAGRE.
`AULA Eat Ree Sme/ke; RRA 50mg/ke AR, AONB.
`MBAR HRA; ZOTRERARFARAY, REFREME,
`FLRER DRA FRATER. WIKRE FRR 3cm,
`2. FAD ARSMRAANSHORE
`-20-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`1) Gly-TB45 TB 4 res
`goLRGR 1) RBLKRARMGRRE, PAW BATRA
`(HZ 10R, SBR SOul/R ), Flext rege (210K, AR
`FARR Sug/50p1/R ), ASeee (Hn 20 R, BRRATAGly-T
`B4 Sug/SOul/R) RRBREH, BAAR,
`2BSAB, MADURA RAMAARMSHAOKA. GRIF:
`Reve (ROKR, BBA), West Reee 3.1540.21,
`Paeext
`foo 3.0740. 18,Gly-T B 4 28 3.1340.11, 402M ARSREF.,
`YRS 5 AW exp we oe 2.3740.18, Pabexp pega 1. 62+0. 13,
`Gly-1 B4#11.1440. 1242ARSHEF.
`LAERER, Cly-TB4 MARRARHIRE (NB 3).
`SL Gly-T B4RERKRGORSH EE RABY BRST BS,
`
`2) Ala-TB45 7B 4 wreak
`weHR 1) FZKRARAGBAE, ATFERPRKHAT
`EXT PR (PBS).
`FAMERS Ream (TB 4), eee (Ala -TB4),
`BARR IK., TB4. Ala -TB4 OBR 100ue/ml,
`AB EHEAMEG URSIN,
`TABS2E RK
`D HARBRM: PBS HHOAS. BS KaeARB,
`478A 1-3; 1B 4 ghAe Ala -TB4 Soa RERMWB, HAY
`1-2 R,
`@ FURS: HAMS] PBS HH 7,341.02 KR, TH4
`245,840.91 KA, Ala -TB4 48H 4.740.83K, Sez ii] oH,
`TB 4 Zafer Ala -TB 4 0% Ay A] AA 4G PBS 2H (P<0. 05), 4
`RIL 7.
`22 RR TB4 Fe Ala -T84 HAA YRHRMGURAHHE
`-21-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`Al. HY, HEA Ala -TR4 HHO RSNABA TB 4 se
`OmRAD: ABRIAG, HREM RRA), RORY:
`A ves} Re 20) 2.340. 31cm, TB 4 20H 1.9+0.23cm, Ala-TB 4 28
`41.740.
`30cm; MATA 4 2hfe Ala -TB4 AM RRAwR
`RRR. RRR, MAM RRRARM, AMRF.
`Aon> ja] roa, TB 4 4hAe Ala -TB 4 484% 0 IRA) PBS 20
`(P<0. 05), 22RILA 8.
`te RA, TB4 fe Ala -TB 4FLAW AgRR OE A
`He, 26/8 Ale-TB 4 4 RR he TB 4>
`
`36%) 5 Gly-TB4. Ala-TB457TB4 3 XR ARMARE HEF
`RRA
`1. Gly-T 645 1B 4 44 rbak
`1). MRBMRA HAE SALA
`ATHY Cly-TR451TA4BAARARMGHERE, BT
`TSHA RGM MRD MRE: ARARBARE.
`JOH2 SAM BAA BBA 1 NYA, RR
`BRAARAKRAREPR,
`30 HUG PBS ADA ZA
`Gly-TB4 5784469, 4AGAHBR, 1LOGRACSA aS
`2, PABIMAAEREAWH, WH ABDRAMAARR,
`BA ARERRA.
`
`2) . AA BARAT AASBRERA
`whLRAGR 1) SGZKARARAGRA SE, PAA EM Ree
`(42010 2, AARRA RAS EMBER 30ul/R, 4K/K),
`FA
`MEX FR 28 (#2010 R, ARBRE 30pl/R, 4R/A), Kosa (
`4210, BARK Gly-TB4 30u1/R, 42K/K) BBB.
`-22-
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`HRARGASH BIR, 10PRALEA HAZ, GAwILM
`JUEP HAD, WMH ABRSMRARR. SHARER
`REL. RF:
`Rae (20K, BBA), Wes eee 4.824012, TB 4 2a
`4.76+0.18, Gly-TB 442 4.8540.16, SHZHMABSREFH.
`43h 4 AA eet Pe Se 4. 8940.12, TB 4407. 03+0.13,6ly-T
`84H 7.62+0.11, Apextesos TB 4 oa, Gly-T844zW ARS
`wee, AS Gly-TB4 AERESRZFH.
`ikUb 4 RRA Cly-T B 4 fe TB 4HB REA RRR
`RA (LB 4). HY, Cly-TB 4stARRR RARAY
`A BF TB 4.
`
`2. Ala-T B45 TB 4 rede
`1). aSawe AERA KE
`ATH Ala-TR45TB4H BMA, BLT SEAR
`YASHE: RKRARRAARE
`JOH8 SABAH RAS) INDRA, RaRTH
`SR RARRARIEPR, MASA EBRKADHTA. LHIA
`AlaTBR45 76436377. PRAT1, SABIHIBRR, 10KPROS
`BAGH, SHMILMPATHRBRWH, WHHABAREG
`A BRAMAARRR. PHARLKRKRABAN,
`2). BAK
`BZLRRARRADRER, AARRERAST WET RY
`(PBS ), FAPEXT RR(TB 4), Re(Ala -TB4), HAR 4K.
`TB 4, Ala-TB4 MHBMREYA 500ug/ml, AASHAMRABE
`BRENT.
`3). RESAER
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`AERRAH: 184 a. Ala -TB 4 Sate PBS ABS
`—AR,MURS) ARERRR(FILA 9A-C). fia, PBS
`PER BRASS ARN, ABD ARERRLAK (A 10A);
`TA4AEBAZARBARN AGH ARELRARLE F,
`AL YAMA) 1-2 BERS (IL 10B); Ala-Th4 2A
`LARBAAN, PRK ARELARSRMYRKRAZER, BTM
`#3) 3-4 BK on (ILA 10C), Znil eA, TB 4 ZnFe Ala
`-TB 422A RERRRLAB PBS 28, Ala-TB 422A RER
`KARA RAF 1B 42a, RAW, TH4 Ae Ala-TR4 AAAS
`AREa 0 ARERAE, HP Ala -TB4 HARER
`RAAAFT TB4,
`
`

`

`WO2007/009359
`
`PCT/CN2006/001679
`
`4Fl) SEK
`
`BRE B44Atzah, Hit yA SEQ IDNO: 6 RRRDI
`1.
`4 Gly-T B 4#e3LAy SEQ ID NO: 5 PRRRRB Hy Ala-T B 4.
`
`—FtSBS), RALAIBR 1 HRYB 4 TE
`
`2.
`Wh
`
`—AHwwe , HEAPFIBR 1 PRAMB 4 TA
`3,
`th, VARIES BA,
`
`PAIR 1 BRR BATE SS RRR.
`4,
`SHELARA. FRAFe BESa Hw ALI.
`
`—FRIGIT LIAM BP RARER AR. ESB eR TG. AR
`5.
`thFa/RILS IANA, LIEDAHRZA BIA IRAE MBA
`BRI PARMAR 4TAY,
`
`6. RARR IMA, APHRLAMRAA.,
`
`25
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`Dy2sBeFBEE(cm) 2 2 a
`
`
`
`BE
`z
`a
`Gly-T B 4
`TB4
`AESB Eb IK
`
`
`—
`
`1
`
`—
`
`_
`
`
`
`Sa tL BUA AT
`m ASLOA 14K
`
`FIG. 1
`
`
`
`MAG FRE
`
`FIG, 2
`
`1/6
`
`35
`_. 30
`S25
`#K 20
`
`@ is
`a 10
`Sn
`
`5 0
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`HOBMGHE
`
`nS
`
`
`
`
`
`FA MRL BEF
`
`
`(on)Sr¢omranawma
`
`
`OGOKEE
`
`
` Gly-T B 4
`
`heCO©
`
`RERIC
`
`2/6
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`WecoSeBe FEBEA
`
`—o— XT R
`—™—-TB4
`\ Ala-TB 4
`
`20
`
`40
`
`FIG. 5
`
`yayT WY Tal (A)
`
`
`
`
`
`
`SEHELL,(%)
`
`
`
`CESAR ERK=EA TB4|HED Ala-TB4
`
`FIG. 6
`
`3/6
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`5 ee TTA
`
`ifTe]CED)OrNWHRom4oO
`
`FIG. 7
`
`PIER BE
`
`AeHEB
`
`TB4
`
`Ala-TB 4
`
`4/6
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679 Bae
`
`i
`
`i
`iyaco
`FIG.9C TB44h, RHGRLA
`
`5/6
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`
`
`
`
`FIG. 10B TB 4 #8, BGR SA
`
`eA
`FIG.10C Ala-TB 448, RHE BSRK
`
`6/6
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`PRUE
`
`<110> JLRS EW RAA REE Fl
`
`<120> ARR BS B 4 AI4EWeEVR a
`
`<130>
`
`TECO60029PCT
`
`<150>
`
`200510083894. 6
`
`<151>
`
`2005-07-15
`
`<150>
`
`200510103293. 7
`
`€151>
`
`2005-09-23
`
`<160>
`
`6
`
`<170> PatentIn version 3.38
`
`<210>
`
`1
`
`<2l1>
`
`48
`
`<212>
`
`PRT
`
`<213> Homo sapiens
`
`<400>
`
`1
`
`Ser Asp Lys Pro Asp Met Ala Glu Ile Glu Lys Phe Asp Lys Ser Lys
`1
`5
`10
`15
`
`Leu Lys Lys Thr Glu Thr Gin Glu Lys Asn Pro Leu Pro Ser Lys Glu
`20
`25
`30
`
`Thr Ile Glu Gln Glu Lys Gln Ala Gly Glu Ser
`35
`40
`
`<210>
`
`2
`
`<211>
`
`<212>
`
`141
`
`DNA
`
`<213> Artificial
`
`<220>
`
`<223> Gly-T-beta 4
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`<400>
`
`2
`
`ggatcceaca aacccgatat gectgagatc gagaaattcg ataagtcgaa actgaagaag
`
`acagagacec aagagaaaaa tccactgcct tccaaagaaa cgattgaaca ggagaagcaa
`
`ecaggecgaat cgtaactcga g
`
`<210>
`
`3
`
`<211>
`
`<212>
`
` 6B4
`
`DNA
`
`<213>
`
`artificial
`
`<220>
`
`<223>
`
`CST
`
`<400>
`
`3
`
`gaattcatet
`
`eccctatact
`
`agettattgg
`
`aaaattaagg
`
`eccttgtgca
`
`acccactcga
`
`cttctttteg
`
`aatatcttga
`
`agaaaaatat
`
`gaagagcatt
`
`tetatgagcg
`
`cgatgaaget
`
`60
`
`120
`
`141
`
`60
`
`120
`
`gataaatggec
`
`gaaacaaaaa
`
`etttgaattg
`
`getttggagt
`
`ttcecaatct
`
`tecttattat
`
`180
`
`attgatggte
`
`atettaaatt
`
`aacacagtct
`
`atggccatca
`
`tacettatat
`
`agctgacaag
`
`tccaaaagag
`
`240
`
`300
`
`cacaacatet
`
`tegetgetts
`
`cetgcagaga
`
`tttcaatgct
`
`tgaaggagcg
`
`gttttggata
`
`ttagatacgeg
`
`tetttcgaga
`
`attgcatata
`
`gtaaagactt
`
`tgaaactctc
`
`aaagttgatt
`
`ttcttagcaa
`
`gctacctgaa
`
`atgctgaaaa
`
`tgttcgaaga
`
`tegtttatet
`
`cataaaacat
`
`atttaaatgeg
`
`tgatcatgta
`
`acccatcctg
`
`acttcatgtt
`
`gtatgacgct
`
`ettgatgttg
`
`ttttatacat
`
`ggacccaatg
`
`tgcectggate
`
`cgttcccaaa
`
`attagtttgt
`
`tttaaaaaac
`
`gtattgaagc
`
`tatcccacaa
`
`attgataagt
`
`acttgaaatc
`
`cagcaagtat
`
`360
`
`420
`
`480
`
`540
`
`600
`
`atagcatggc
`
`ctttgcageg
`
`ctggcaagec
`
`acetttgetg
`
`gtggcgacca
`
`tectccaaaa
`
`660
`
`tcggatctgg
`
`ttccgcgteg
`
`atcc
`
`684
`
`<210>
`
`4
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`<211>
`
`153
`
`<212>
`
`DNA
`
`<213> artificial
`
`<220>
`
`<223> Ala-T-beta4
`
`<400>
`
`4
`
`ggatcccctc gagcttctga caaacccgat atgegctgaga tcgagaaatt ceataagtcg
`
`60
`
`aaactgaaga apacagagac gcaagagaaa aatccactge cttccaaaga aacgattgaa
`
`120
`
`caggagaage aagcaggcga atcgtaactc gag
`
`153
`
`<210>
`
`5
`
`<211>
`
`44
`
`<212>
`
`PRT
`
`<213> artificial
`
`<220>
`
`<223> Ala-T-beta 4
`
`<400>
`
`5
`
`Ala Ser Asp Lys Pro Asp Met Ala Glu Ile Glu Lys Phe Asp Lys Ser
`1
`5
`10
`15
`
`Lys Leu Lys Lys Thr Glu Thr Gln Glu Lys Asn Pro Leu Pro Ser Lys
`20
`25
`30
`
`Glu Thr Ile Glu Gln Glu Lys Gln Ala Gly Glu Ser
`35
`40
`
`<210>
`
`6
`
`<2li> 44
`
`<212>
`
`PRT
`
`<213> artificial
`
`<220>
`
`<223> Gly-T-beta 4
`
`

`

`WO 2007/009359
`
`PCT/CN2006/001679
`
`<400>
`
`6
`
`Gly Ser Asp Lys Pro Asp Met Ala Glu Ile Glu Lys Phe Asp Lys Ser
`1
`5
`10
`15
`
`Lys Leu Lys Lys Thr Glu Thr Gln Glu Lys Asn Pro Leu Pro Ser Lys
`20
`25
`30
`
`Glu Thr Ile Glu Gln Glu Lys Gin Ala Gly Glu Ser
`35
`40
`
`

`

`PCT/CN2006/001679
`
`KFEORAY ti BA
`CAA132)
`
`GRIN]_2006 #0705
`CAPPER CUSERT)
`EZeeenmatC1|
`
`
`DAH EA PITRE REN CORUAAEA Pais eB hi eM)
`
`
`EAPC UU] CUBED
`POS AB ate(SURE, Plbd: “ARIAS -S”)
`
`
`
`1-25 47PTBAY BRIAEyiet
`AMUABH_9 TH. #
`BS AREER C1 |
`B.RE
`rhBekaRILSSIBE
`spaenteee
`"China General Microbiological Culture Collection Center (CGMCC)
`RERL
`CALE IGARS0 IS)
`
`cb IGAE RKPet 13 EB, PREBLE ETSEBT (100080)
`
`
`
`chy 523eS
`
`HE ir ates
`
`C) AR EAAE i is—HS
`
`[] Beal Tr A
`
`PCT/RO/134 #% (1998 4E 7A, 200442 1 AFAR)
`
`

`

` HABREEARES 1BC060029PCT
`
`Bm eeS PCT/CN2006 /00 1679
`
`PCT/CN2006/001679
`
`FFtre0ek BY Ub BA
`(4am 13 2)
`
`ASU BSS_9 Tl, Fo__21-25 FTATR RAEURteaaA
`B. Ria
`ES REMMTA CO]
`aaafe
`FF BeBsPLAGE
`aie China General Microbiological Culture Collection Center (CGMCC) |
`FeaALIA
`(ALFRMD BSSA BIZ )
`IDR TER PoE 13S, PHBREDHAT (100080)
`
`TRIXESA 20064 07 A 05 A __|PRIS _CGMCC No.1751 _
`
`BZA
`CARERS CUSED)
`
`
`
`
`DAWHEA TATEER ER CORI MABDH BPAIEe BED
`
`HAL EA
`
`E.4bFauid CUBR)
`
`
`
`
`PAAR REE CSU HRAUSSSy, Bilin.
`
`“GRA sae”)
`
`
`HH S23 Fade
`
`tH El Bas aS
`
`
`(] AW BAHHit Yal
`
`
`Fl BaneaeBiAST EHF
`
`
`PCT/RO/134 #% (1998 4E 7A, 200442 1 ABH)
`
`

`

`
`
`International application No.
`INTERNATIONAL SEARCH REPORT
`
`
`A. CLASSIFICATION OF SUBJECT MATTER
`
`
`
`According to International Patent Classification (IPC)or to both national classification and IPC
`
`
`
`
`Documentation searched other than minimum documentationto the extent that such documents are includedin the fields searched
`
`
`
`
`
`
`PCT/CN2006/001679
`
`See extra sheet
`
`B.
`
`FIELDS SEARCHED
`
`Minimum documentation searched(classification system followed by classification symbols)
`
`IPC’: CO7K, A61K, A6G1P
`
`Electronic data base consulted during the international search (nameof data base and, where practicable, search terms used)
`
`WPI, EPODOC, PAJ, WEB OF KNOWLEDGE, ELSEVIER, GENBANK, CPRS, CNKI (thymosin beta4, tbeta4, variant, mutant,
`
`analog, analogue, derivate, derivative, SEQ ID NO:5-6etal.)
`
`C. DOCUMENTS CONSIDERED TO BE RELEVANT
`
`Category*
`
`Citation of document, with indication, where appropriate, of the relevant passages
`
`Relevant to claim No.
`
`
`
`A
`
`A
`
`A
`
`*
`
`“A”
`
`“R”
`
`“L”
`
`
`
`
`US,A,4297276 (UNIV WASHINGTON GEORG) 27. Oct. 1981 (27.10.1981), see
`the whole document
`
`US,A,4395404 (UNIV WASHINGTON GEORG)26. Jul. 1983 (26.07.1983),see
`the whole document
`
`1-4
`
`1-4
`
`|
`
`Simenel C et al., “Structural requirements for thymosin (4 in its contact with actin ”,}| 1-4
`Eur. J. Biochem, Vol.267, 2000, pages 3530-3538, see the whole document
`
`Further documentsarelisted in the continuation of Box C.
`
`See patent family annex.
`
`Special categories of cited documents:
`
`“T”
`
`document defining the general state of the art which is not
`considered to beofparticular relevance
`
`later docum