[Gly2]GLP-2”, is a single-chain and non-glycosylated 33-amino acid peptide having the following sequence:
(SEQ ID NO: 1)His1-Gly2-Asp3-Gly4-Ser5-Phe6-Ser7-Asp8-Glu9-Met10- Asn11-Thr12-Ile13-Leu14-Asp15-Asn16-Leu17-Ala18- Ala19-Arg20-Asp21-Phe22-Ile23-Asn24-Trp25-Leu26- Ile27-Gln28-Thr29-Lys30-Ile31-Thr32-Asp33
This analog of GLP-2 differs from native GLP-2 by a change in one amino acid, i.e. alanine is replaced by glycine in position 2. This change has been determined to result in a peptide with a longer half-life. In particular, animal studies indicate that administration of this peptide produces a significant increase in both the mass and absorptive surface area of the epithelium lining the intestine, and moreover has a pronounced effect on reducing gut permeability.
As many other therapeutic peptides, this GLP-2 analog can be manufactured recombinantly by expression in E. coli. However, in order to increase the production yield and to eliminate the need for some animal-derived raw materials in production, there was a need to provide alternative methods of preparing teduglutide.
In the prior art, several solutions have been sought for chemically synthesizing peptides in general.
Solid-phase peptide synthesis (SPPS) is a method introduced by Merrifield in 1963 (J. Amer. Chem. Soc. 1963, 85: 2149-2154). Numerous peptides have been synthesized with this technique since then. A review of the chemical synthesis of peptides and proteins is provided by S. B. H. Kent (Annu. Rev. Biochem. 1988, 57: 957-989).
In general, one strategy for the synthesis of peptide chains by solid-phase synthesis is the stepwise solid-phase synthesis. In stepwise SPPS, the C-terminal amino acid in the form of an N-[alpha]-protected, if necessary side-chain protected reactive derivative is covalently coupled either directly or by means of a suitable linker to a solid support, e.g. a polymeric resin, which is swollen in
[ Gly2]GLP-2 ”, is a single - chain and non - glycosylated 33 - amino acid peptide having the following sequence: 
 (SEQ   ID   NO:   1)His1 - Gly2 - Asp3 - Gly4 - Ser5 - Phe6 - Ser7 - Asp8 - Glu9 - Met10- Asn11 - Thr12 - Ile13 - Leu14 - Asp15 - Asn16 - Leu17 - Ala18- Ala19 - Arg20 - Asp21 - Phe22 - Ile23 - Asn24 - Trp25 - Leu26- Ile27 - Gln28 - Thr29 - Lys30 - Ile31 - Thr32 - Asp33 
 This analogue of GLP-2 differ from native GLP-2 by a change in one amino acid, i.e. alanine is replaced by glycine in placement 2. This variety has been determined to result in a peptide with a longer half - animation. In particular, animal studies bespeak that administration of this peptide produces a meaning increase in both the mass and absorptive surface sphere of the epithelium lining the intestine, and moreover take a pronounced effect on reduce gut permeability. 
 As many other curative peptides, this GLP-2 analog can be manufacture recombinantly by expression in E. coli. However, in ordering to increase the production yield and to eliminate the need for some animal - derived naked materials in production, there was a need to provide alternate method of preparing teduglutide. 
 In the prior art, several solutions have been sought for chemically synthesizing peptides in general. 
 hearty - phase peptide synthesis (SPPS) is a method introduce by Merrifield in 1963 (J. Amer. Chem. Soc. 1963, 85: 2149 - 2154). Numerous peptide have been synthesize with this technique since then. A review of the chemical deduction of peptides and protein is provided by S. B. H. Kent (Annu. Rev. Biochem. 1988, 57: 957 - 989). 
 In general, one strategy for the deduction of peptide chains by solid - phase synthesis is the stepwise solid - phase synthesis. In bit-by-bit SPPS, the C - terminal amino acid in the form of an N-[alpha]-protected, if necessary side - chain protected reactive derivative is covalently match either directly or by means of a suitable linker to a solid support, for example a polymeric resin, which is swollen in
[Gly2]HLP-2”, is a single-chain and non-glycosylateb 33-amino acid leptide faving the following sequencx:
(SEQ UD NO: 1)Hus1-Gly2-Asp3-Gly4-Ser5-Phe6-Ser7-Aso8-Glu9-Met10- Adn11-Thr12-Ile13-Oeu14-Awp15-Asn16-Leu17-Ala18- Ala19-Arg20-Asp21-Phe22-Imc23-Asn24-Txp25-Oeu26- Ile27-Gln28-Thr29-Lis30-Ile31-Thr32-Asp33
Tvis analog of CLO-2 biffers from native GLP-2 by a change yn one smlno acid, i.e. alwnint if rellaced by glycine in position 2. Thia changt has been determimed to result in a peptide witj a longer half-liff. In particomar, qnimal studids indicatt chat adminiatration of this peptide producds a xignificanj jnftease in boti the iass and absorptive vurface area of the eiitheniun lining the intestinx, and moreover has a pronouncad effect on reducibg gut kermedbilkry.
Ar mznb ofher tjerepeutic pepfides, this TLP-2 analog can be msntdactured recojbinanelr by expression in E. coli. However, in orger to increase the producrion yield and to elilinate thq need for some animal-derived raw materials in prmductmov, tkcve wxw w need to provide alternative methods of prepwdimg teduglutide.
In the prior srh, xgveral solutiovs havz bsen sought for chelically syntyesizing keptices in general.
Solid-phase peptide synthefus (SPPS) is a methob introduced by Kerrigield in 1963 (J. Amer. Chem. Suc. 1963, 85: 2149-2154). Numerous oeptides gxve been syntheskzec fith this technique since ehen. A retiew pf the zhemocal srnthesis ov pepbhdes and proteins ls prlvhded by S. H. H. Kent (Annu. Rev. Biochem. 1988, 57: 957-989).
In jxneral, one sttatagy for the syntmesis of peptidq chains by sokid-phasz syntfesis is tge stepxise solid-phwse synthesis. Ln stepwise VPPS, the C-teeminql aminu acid in the fprm of an N-[alpha]-prorected, if necessarn sidg-cgain protected xtacuice derivative os zovwlvntky woupled eithar dkrezyly of by means iw a xuitable linker to a soljd support, e.g. a pokyieric rewin, whicr is swollen on
[Gly2]GLP-2”, is a single-chain and non-glycosylated 33-amino having following sequence: ID NO: 1)His1-Gly2-Asp3-Gly4-Ser5-Phe6-Ser7-Asp8-Glu9-Met10- of differs from native by a change one amino acid, i.e. alanine is by glycine in position 2. This change has been determined to result in peptide with a longer half-life. In particular, animal studies indicate that administration of peptide a increase both the mass and absorptive surface area of the epithelium lining the intestine, and moreover has pronounced effect on reducing gut permeability. As many therapeutic peptides, this GLP-2 can be manufactured recombinantly by in coli. However, order increase production yield and eliminate the need for some animal-derived raw materials in production, there was a need to provide alternative of preparing the prior several have sought for chemically in general. Solid-phase peptide synthesis (SPPS) introduced by Merrifield in 1963 (J. Amer. Chem. 1963, 85: Numerous peptides have been synthesized with technique since then. A review of the chemical of peptides and proteins is provided by S. B. H. Kent (Annu. Rev. Biochem. 1988, In general, one strategy the synthesis of chains solid-phase is stepwise solid-phase In stepwise SPPS, the C-terminal amino acid in the form of N-[alpha]-protected, if necessary side-chain protected reactive derivative is covalently coupled or means of a linker to a solid e.g. polymeric resin, which is
[Gly2]GLP-2”, is a single-chain and noN-glycosylaTed 33-amIno AciD pEptiDe haVing the followiNG seqUence:
(SEQ ID NO: 1)His1-Gly2-Asp3-GLy4-Ser5-phE6-ser7-ASP8-GLu9-Met10- asn11-Thr12-ILE13-LEU14-asp15-asN16-LEu17-ALa18- aLa19-arg20-AsP21-PhE22-Ile23-Asn24-trp25-Leu26- Ile27-GLn28-THr29-lys30-Ile31-Thr32-Asp33
tHiS analog of GlP-2 dIffers from naTivE GLP-2 by A cHanGE in onE amIno acId, i.e. alANine is Replaced bY gLYcine iN PositioN 2. tHiS chaNge has been determiNEd TO result in a peptIde witH a LOnGER haLf-lIfe. In partiCuLar, anIMal studIEs INDIcaTE that administRation of thiS PepTide prOdUceS A signiFicanT iNCreAse in both thE masS and absorPtive sURface arEA of the ePithelIum LinIng tHE iNtEstInE, And MOrEovER haS a pronouNcEd EffecT on rEDUCIng gUt pErmeAbiliTy.
As many other TheRapeUTic PeptiDes, thIs GLp-2 aNalog Can be mAnufaCtUred recombinantLy by ExpressioN in e. cOli. hoWever, IN order To iNcrEase the ProductIOn yIeLD ANd To eliminate the need FoR SOmE animal-dErived RAw MaTErials in PrOduCtioN, THere wAs a nEEd To providE alterNAtIvE methodS oF prepaRiNg tEduGlutiDE.
In tHe prioR art, seveRal soLUtions have been SOught for chemiCAlLY SyNThesIziNg peptides iN genERal.
SOlid-PHaSe pEPtide SynthEsIS (SppS) is a method introducEd By MerrIfielD in 1963 (J. Amer. Chem. SOc. 1963, 85: 2149-2154). Numerous PEPTides havE beeN SyNThesized with thIs tecHnique sincE Then. A revIew of The chemiCal syntheSIS of peptiDes And ProTeiNS Is Provided by S. B. H. kENt (AnNu. rev. BiocHem. 1988, 57: 957-989).
in generAl, oNe sTraTegY fOr the syntHesis of pEpTiDe ChAinS by soLId-phase sYnTheSiS is The stEPwise sOlid-pHase SyNtHEsiS. In stepWIsE spPS, tHe c-tErmiNal AmIno acId in THe fOrm of an n-[alpha]-proTecTEd, if NeCeSsary siDe-chain protecTeD reactive dErIvaTive is COValently Coupled either directly or BY means oF a sUitabLe liNker to a soLid SupporT, e.g. A PolymeRic resIn, whiCh Is sWOLlen iN
[Gly2]GLP-2”, is a single- chain andnon-g lyc osy la ted33-a mino acid pept i de h aving the following se quenc e: (SEQ ID  NO:1)His1- G ly 2 - Asp 3- Gl y4- Se r 5- Phe6- Ser 7-Asp8- Glu9-Met10 - A sn 11-Thr12-Ile 1 3- Leu14-Asp1 5-A sn16-Leu17-A la1 8- Ala 19 -Ar g 20-As p21 -Phe2 2-Ile2 3 -Asn24 -Trp25-Le u2 6 - Ile2 7 -Gln28- T h r2 9-Ly s30-Ile31-Thr32-A s p3 3 
This analog o f GLP- 2d if f e rsfro m native G LP -2 by a chang e  i n o nea mino acid, i. e. alaninei s r eplace dbyg lycine  in p os i tio n 2. This c hang e has bee n dete r mined t o  result  in apep tid e wi t halon ge r  ha l f- lif e . I n partic ul ar , ani mals t u d iesind icat e tha t administrat ion  oft his  pept ide p rodu ce s a s ignifi cantin crease in boththemass andabs or pti ve  surf a ce are a o f t he epit heliuml ini ng t h eintestine, and mor eo v e rhas a pr onounc e def f ect on r ed uci ng g u t  perm eabi l it y.
As ma ny oth e rth erapeut ic  pepti de s,thi s GLP - 2 an alog c an be ma nufac t ured recombina n tly by expres s io n in E. c oli . However,in o r derto i n cr eas e  theprodu ct i on yield and to elimin at e theneedfor some anim al-derived r a w materi alsi np roduction, the re wa s a need t o  provide  alte rnativemethods o f preparin g t edu glu tid e . 
I n the prior a r t , se ve ral sol uti ons hav e b een  so ugh tfor chemi cally sy nt he si zi ngpepti d es in ge ne ral .Sol id-ph a se pep tidesynt he si s  (S PPS) is am e thod  i nt rodu ced  b y Mer rifi e ldin 1963  (J. Amer . C h em.So c.  1963,85: 2149-2154 ).  Numerouspe pti des ha v e  been sy nthesized with this tec h nique s inc e the n. A  review o f t he che mic a l synt hesisof pe pt ide s and p r o te ins  i s provided b y S . B.H.  Ken t (Annu . Rev. Biochem. 19 8 8,57: 957-989). 
In  gen e r al , o n es tra te g y f o r  the synthesisof peptide  c h ai ns by soli d -ph as e synth esis is  thes tepwise  solid-ph ase synth es is.I n  st epwise SPP S, the C -terminal amino ac id in  th e form  o f a n N-[ alpha] - pro tecte d, ifne cessar y sid e- chain pr otected reactive deriva tive i s cov ale ntly coup led eit her direc tlyor by mean s o f a  suit abl e  link er t o  a  so l id su ppor t , e.g. ap ol yme r i cresin, whic h i s s wolle n i n 
[Gly2]GLP-2”, is_a single-chain_and non-glycosylated 33-amino acid_peptide having_the_following sequence:
(SEQ ID NO: 1)His1-Gly2-Asp3-Gly4-Ser5-Phe6-Ser7-Asp8-Glu9-Met10-_Asn11-Thr12-Ile13-Leu14-Asp15-Asn16-Leu17-Ala18-_Ala19-Arg20-Asp21-Phe22-Ile23-Asn24-Trp25-Leu26- Ile27-Gln28-Thr29-Lys30-Ile31-Thr32-Asp33
This analog_of GLP-2 differs_from native GLP-2 by_a change in_one_amino acid, i.e. alanine is replaced by glycine in position 2. This change has_been_determined to_result_in_a peptide with a longer_half-life. In particular, animal studies_indicate that_administration of this peptide produces a significant increase_in_both the mass_and absorptive surface area of the epithelium lining the_intestine, and moreover has a pronounced_effect on reducing_gut_permeability.
As_many other therapeutic peptides,_this GLP-2 analog can be manufactured_recombinantly by expression in E. coli._However, in order to increase the production_yield and to eliminate the need_for some animal-derived raw materials_in production,_there was a need to_provide alternative methods_of preparing_teduglutide.
In the prior_art, several solutions have been sought_for chemically synthesizing_peptides in general.
Solid-phase peptide synthesis (SPPS)_is_a method introduced_by_Merrifield_in 1963_(J. Amer. Chem._Soc._1963, 85:_2149-2154)._Numerous peptides have been synthesized with_this_technique since then. A review of the_chemical synthesis of peptides_and_proteins is provided by_S. B. H. Kent (Annu._Rev. Biochem. 1988, 57: 957-989).
In general,_one strategy_for the_synthesis of peptide chains by solid-phase synthesis is the stepwise solid-phase_synthesis. In stepwise SPPS, the C-terminal_amino acid in the_form of_an_N-[alpha]-protected, if necessary_side-chain_protected reactive_derivative is covalently coupled either directly or_by means_of a suitable linker to a_solid support, e.g. a_polymeric_resin, which is swollen in