Free Declaration - District Court of Delaware - Delaware

Case1:04-cv—00884-SLR D0cument71-22 Filed 07/17/2006 Page10f4
E h `b ’t 1 6

, Case 1 :04-cv—00884-SLR Docu ment 71 -22 Filed 07/17/2006 Page 2 of 4
United States Patent Office 3·“”·""
Patented Feb. 24, 1970

1 2 \ Q
which latter is then oxidized, I bl `th lass ® '
3 491 491 ferrieyanide in an aqueous soltlghr?-:t p’Hwl5.5·?I° to E
[bg pqlypgpgjdc gf Formula L m
Nng::¤z*‘;[:.‘g:'“ x::;'.’*·“:é"W"?Q:'E';E‘h‘: The protected ocrapeptide derivative ot Formula. II can , q
alcvuiska Altarlenue V¢d.,Pragne, Cz$toslovalda 6 ii °!::‘°d W °$h“’ P'°“d°"’_’“°h ” *l° °°¤"°¤· _
No nam;. aura sqrt. 14, rm, so. No. ssv,m "°”° "' P'°P°¤'*°¤ °’ P°'¥P°P'!*’°°· Th 9°¤*P°*&¤"
\ °*·*·¤ ·*t,·*‘t*t§·;‘;.·:**·;;·,$:;t*;··*···*·*~ 1‘.‘t.1?‘1i."i.§'*“'i’."".‘§i." "’ ‘ ""‘";it "".§‘°"‘ §‘*'i‘
ep , e mo run groupasw as eear xyic _
ht. Cl. CD7: I0.!/52; Atiilt 17/00; C0‘ld 93/35 terminal sroup, or by coupling of lypeptides eontainio
.s. so-1u.s · · - °° - ‘
U CI. 2 I Claim yu :,l;e :r}trn;hact'dc sequexes oIl;hetio;1fpgypeptide
_,__.____ . e rnpora pro etron e ammo
lib e '·
l ABSTRACT OF THE DISCLGSURE §ir::l:io:;‘:aPy°b;‘1;red¥:;rmy;g gugzahzdegfsdgrtigxs
· at are conventions in e synthetic he ' t t' l - .
b,"*"'**",P_1.,'_,b¢_¤`n_A_s¤ c"_,m.D_An_m,_NH_ 15 peptides. Eg., a r-butyloxycarbooyl, o·:itrot;¤:e'r·i’yl¢;ull,;,n;l
l 1 2 a_ t t e 1 t t tn or tntyl group can be used for blocking ot the amino
t ~·¤=~*grg*g B #¤·=·¤=¤·¤¤*¤¤¤¤¤*¤ ¤=*¤ ¢M=»>> ¤· ¤¤=*- fil’“§§;.2..i$I.’l;r"fr'L"1¥.i‘rr$r}’,i°iZ$‘Z.'}."`£¥} E'2’.l’.‘Lt’3Z
t..,.,.S;$‘2“Fit’§.¥.’.§i£;Li’i.§‘. . tn. ..s.,en. 2,, *_;;:tg,··g;·**··¤~" mu tu °·= ¤····*¤=···=·· tt ¤·= ¤~·····
anudimuc aww' The jrrotective groups are then removed ln the usual
—................ manner n one stage or successively. Oxidation ot the re·
duced form ot` the polypeptide of Formula IH to the cyclic
BACKGROUND OF THE INVENTION 2_ pzytgeptide ot Fotinul: I may accomplished by meth-
Compounds exhibiting a high and specitie antidiuretic ° 9 mivm P"` s°’ M nm°°°’ m m °q°°°“’ ’?l'"i°° °r ‘ ‘
activity are important in the therapy of diabetes insipidus. ln; °f `;°m {ud ‘°,v°°“.mk°m° mm wai"`
Such compounds are well known and represent structural I in nl Y' um °"°w°P$ m°'h°d.'° "°Fd‘ c°nq°"m’°"
modihcatioos of the vasopressin molecule. The most suit.— °l p` I1? ':s"°*’;:°P°°P'°"yI °m°“d‘ ""u’ 'Y'°"“° “"lh'
able properties have been observed in the case of l·de- an Y Nin, °° ml ° ’° in "m°p°"°d p`b°n?Ylm°"°P‘°'
am;m_Phc;_Ar&._vI$°pr“si¤ (ch me uhh bd°w)_ proprooyl-L-tyrosine methyl ester. tty the action of hydra-
SUMMARY OF INV ON ;•neltzdrate, the latte;]c•;lmpound ra convened to the so
¤ air r wsse urtreport ydrazide and is then coupled ‘
It is_ort ohiocfof the present invention to provide for WM L'PlE°¤Zm*§*'“° mmwl ‘“"'· Inf '°*“m¤8 dl¤¢P*ld¤
an anudrurerre that has a higher degree of activity and °“°" d°f""*"’° '* ‘{°“d°¤”d· ‘¤‘E"' ln 'M {Wm °‘ {M
is more speeitic in its action than the just named vasopres- as um'- "”"h '··‘!1‘"·“'!""Y'*1**‘P“'i*¤“'Yl‘S·l>°¤ZYl·L‘¤Y*i°¥¤¤
sin cm·,p°¤mg_ methyl ester to give the hitherto likewise unreported
1-;,;, is ,,,,,,,,,;;,;,,,1 by , p,yyp,p,;dc nf lh, Fun p - benzylrnereaptopropronyt-L-tyrosyl·L;plrenyIalanyl·L-
mgh { glutar¤t¤yl·L·asparaglnyl·S-benzyl-L·cystetne anethyl ester
_ w wlgclx ts coupled (by the apde synthesis) wrth L-proly|·
~*;»»¢;·-rt--¤;»-q··-¤';··¤·;~-¤-;·-¤;··~¤· ,,, £Z..;§§’.‘i}§£'E"‘.¥J§Zi§2L‘2“.?é’.i;‘.‘I,‘I$.‘?ti‘3.*$§‘.‘£3.°£?F3}?
trtula 11. The protective groups are removed by reduction
wherein Mep is bmercaptopropionic acid (Mep) at posi- Wim ‘°dlP¤`l i¤ iiqvid °'¤mQ¤i*_*¤d i{l¤ Imlmllk |’¤d¤¤¤d
gm, [ and D.a";,,;,,, is ,, Fashion 8_ 45 product rs converted by oxrdutron with potassium terri-
ma mmppund ohh; gnmgcn Im ibm, and spuggc gyantde t? n new cycltc biologically active disulfide of
antidiuretic activity. ¤¤¤¤ 1 ·_ _ _ _ _
¤uscmmé>l):B?)§=)IrN}EmN1t;s22Fcnn2¤ ,...E`°;;{Zii§°.:.§igi¥.:.i¢F:$i3 mst::i;Bt:elx?
than t e nature y occurring arginine-vasopressin. The cli-
— nre compound of me invention is presently rome ‘° =¤lf·¤= 1 ¤i¤¤r¤ irvm ¤rs¤¤in=-v¤=¤¤r===i¤ bv ¤h¤ r¤¤==¤¤=
by condensation ot pbenzylmet-¤ptop;opi¤¤y|.L4y;¤_;y]. of pmercaptoprcplonie acid at position l instead of
L-pheuyalanyl - L - ;lutaminyl·L-asparaginyt-smeuyt-L. cysteine and by the occurrence of D-arginine at position
cysteine azide with L-pr¤Iy[·N°-tasyl-D.argi¤ylglyg;in. B instead ot !.·arginirte. The polypeptide of Formula I
amide to give nre protector octapeptide uertvsrtve or me which my he d==is¤¤¤=¤ "1—¤=¤¤¢¤¤-¤—¤·=r¤l¤i¤= Iollowing Formula Il 55 pressi¤" represe¤ts—due to its biological activity-a valu- ’
_ _ _ _ _ able dnrg or the regulation of polyuria and moderation of
“°"""’"'"’°°'*"“°""§Z1&?i%»..~r, rm ¤°¥.;’:°;.§:.§:*:,§; ‘;§.‘;"£’.f."i$1’§‘ii..‘“?.‘3.i§'“.i‘ t r r.
IIB D C
wherein Rl and R’ are protective groups. e.g.. R' may be invention with the urgiuinegvasopresin. P
benzyl and R’ maybea p-toluenesulfonylgrnup. This step °° For the determination of the antidiuretic activity the
rs followed by removal of the protccttve groups of For- method of Sawyer (sqwyq, W_ H, guqpuimtngy 53. 594 r
m¤l¤_ U by { f=<§¤¤§¤¤¤ f¢¤¤¤§>¤. ¢·B;. by ¤¤= ¤¢i¤¤¤ ¤f Mt (195B)) ss tundiued by Ptiska et aL (Plidra, V.; Ryehlik,
°2°°&:i;”§;°;:l:;‘¤:3:¥iII'“m°m* *° ¥"’” *1** Y‘d““d P°lY‘ I., Acta Qndocrinol, 54, l29 ( 1967)) was used: experi-
F P 65 mental animals: Wistar nt, female, weight l$0—210 g. The
Mep(Sl-I)·’!‘yr-Phe-Gln-Asn—(.§·s(Sli)-Pro-D. prcssor activity was determined by the Laodgrcbe method
Ar;·Gly-NH; (Ill) (Landgrebe, F. W.; Macaulay, M. H.; Waring, H., Pro-

Case 1 :04-cv—00884-SLR Document 71 -22 Flled 07/17/2006 Page 3 of 4 T Vaeael . .
_
3,497,491
3 4 1
ceedirlu of the Royal Soc. Edinburg B 62, 202 (1946)- _ ~ _ _ .
experimental animals: male Wistar rates, weight 182; A B°°zylm°ru’wPrz;2£ I;s:g°°Yl°I’Ph°"yl°l°m°° “
250 . ·
l nm.: p-BemyImereaptopr¤pi¤ayl—L-tyrosine hydnzide ($.6 A. .1
5 g.) is dissolved in a mixture of dimethyltormamide ($0
___;‘__"*"_L'j_____ ml.) and concentrated hydrochloric acid {4 TL). Ee ·
E . ants Smclhnlky solution is treated under stirring and cooling at ·· ' .
°*°*•“°· "*$°°’· °'"*: c.), will sodium nitrite (1.03 ny in water rz ml.), The _
Compound l.l!J¤g. LV Im!. aetteu _ _ _
pH of the reaction mixture is then admsted to a value of
*‘¤‘°”‘{‘f{tf_‘f_':fffj’j____ ,,_m&,,,, 1, mu, m from 6 to 7 with N·ethylpiperidi¤e, and a solution oi i ·¤
1-Eamnnrhsltiu-me Z lrphenylalnnine methyl ester (2.7 g.) in dinrethylform- .
amide gs nn.) is added. me reaction mixture is left-
,j,;.’;-é;¤_?~‘i§%‘g§¥§§¥§5T¤*¢“~“’·’i·i?#·l“»¤‘»“¥‘»”il~~¤· ?§;7,?‘.?....."°$‘f.‘$°l.“2l.2..‘§;.i°‘.£.f§§.’.I`.'€‘i£.§§.'»...°°*" ‘
·.lr¤;aeatn. EL., Balnnans. 1.1-: Kclv- ¤htm.Aet¤••.¤¤t¤¤¤l N press¤re._'l'he olly residue is trlturated with watc:jith§ I _
.· ‘¤¤· ¤¤¤v¤=¤¤¤= 1 ¤f ¤== i¤~=¤¤¤¤ ·¤·v ¤= ¤==·* e= ¤¤= i{Z.i.°Z§£ZY·KI°2`n'Z’L¤i?'{»‘.T»`i° §T1.°';°'?.?‘$£'.'E¥'¥.?}....‘§» A
f"° bm °' i'·‘ u" hm °¢ i“ mls Wim i”°'g°°‘° °' merceptoptopionyl-L·lyrosyl·l'.»pheliylala¤ine methyl es- .
nrganie acids, without or with additives auch as stabilizers. m MJ, BLU;. C. awk,] mmm,. ‘
f‘· preservatives, sweetening agents, aromatic compounds, 20 ‘ ’ " '
Havering agents or detergents to obtain the suitable form, [¢]¤”·—l7-$:(h5‘ V g
asugesired, io; theugxenlezl. ilféglil. l¤lT¤¤;;§.a§gk (e. 0.5, dimethylformamide), For a product. ¤bl¤l¤¢d V
Cll ¤0l.lS, Ul lm I! V _ I I ’ 687 · ld b |· _bc_“z]nm m in [
‘ ’1'h¤¤. =-n·. hy4r¤=1¤1¤¤i¤ ¤¥i¤» r>¤¤=r¤¤¤¢ wd. =¤¤ ¤¤¤= iii; urelinmaenyh(2:;]:-`:.ii1fr..ly..»yli{i;i.e.g1:im?ie
mid <=¤¤ be M ¤= i¤¤r¤¤{¤i¤ ¤¢i¤=. ¤¤=¤¤ Mia. ¤i¤i¤ ¤¤i¤· M methyl ester ne renewing vane. were given an ue 1itera·
Blld lBI'l&I’l¤ ICN IS DYKIIIIC Milli, IDG, huhlll IS 1 mIl¤· _ Imc: M_P_ 147_]48¤ C, [‘]¤”_l75=x• (c' L4,. dI_
Iii! hal/l¤8 ¤¤ ¤¤ld$¢ !“¤¢!i°¤· ¥“*’lh“m°*'°¤ much- I"' methylformamide), e£.Huguenin R.L.;·Boissortna=. R.A= ·
tore, natural or hydrogenated oils, talc, and glycerol ¢3¤ Helv. Chim. Acta 49, 695 (l9d6). l
be used as further additives. .
The preferred way of administration is subcutaneous so P'B°l¤YY¤¢F¢*Pl°P¤°Pl°¤)'|·I;·!Yf¤!!/I-I··Pl¤¤¤Yl¤}¤¤l¤¤ ·
but the drug gan nlsonhe uxd in¤·a:1as:‘il|,:pplieatiltl1:z|; hYd¤’¤¤d¢
Thedoseisa outs- ng. g. ayin e tease. A ’t¤ I -benzylm t ` l-Ll M,.
(—. dose iozgntranasal administration is approximately 200 phnnylillfxwglyz :1c£y1¢¤cr(i;i;pB$¥r:`€:‘;‘:i°[({531LL ,
ns-/ka- Y- " and¤0%h ' hdt 6 l.' tl df 2
· The invention will hejurther illustrated by the follow- 35 hours. The zdrgtruzls wgie: gegarxgogt iggategrby
ang eguampges, alllsicgglétcfnzzafgtlkd l}J¢l’¤l¤. All |¢¤· eppiyinuzmgzu, washed while on the filter with t¤elh< ‘
pers es re us e . ano a e an recrystallized from dimethylform.
gxAMp;_Eg amide-water. Yield. 10.1 g. (72%) ot pebenzylmereapto-
_ _ proplonyl·L-tyrosine · L · phenylalanine hydrazlde. M.P.
p·Benzylmereapt¤propionyl·L~tyrosme methyl ester W ;4g.,;;q• C; Dplgm rgunoug [dbi. ._2;_y.._.0_;• {,_
paenzylmemaptnnrnnienie acid (9.2 g.) is convened 0-55. dimelhvlformnmidol. Recorded (see the 1=r=¢¢dl¤e
gylresetirig miith thiogyl tihltaride {E :he norrespgndgg P¤’¤8¤Ph)= M·`P· 2$l·~2$3' C.
or`e. eatteris issove ine yaeetate(2 m.. . g _ _ __ _-
and the solution. under vigorous stirring and cooling 5gm;%:;£gI:;:£::£yI_;b=g;??_:_;,`s$:;:yl::g;l .
(ll' C.), is added to a mixture of l,·tyrosi¤e methyl ester cm,
hydrochloride (10.9 g.), water (50 ml.), and ethyl acetate 45 _ _
no mi.); ne pH nin. ofthe ...nl¤.. mixture is nn in ¤ - *§·=·¤v¤¤·¤r¤¤¤·¤p·¤¤·¤¤v¤·¤»¢v¤·z=v¤-¤-·¤¤¤=¤v¤el=¤·¤=
the mild alkaline region by additions of sodium bloat- hYd'ul{’° (89 EJ is ¤¤¤<}=¤$¤d Willi I-·BI¤l¤l¤l¤Yl·I-·
bonate. The ethyl acetate layer is separated, dried, eva- ¤$P¤{1I¤¤1/l·$·b¢¤¢yl·L·¤ytteme methyl ester (7.2 g.) ae-
_ poratcd under stlhatmospherie pressure and the residue is 50 °°¥dl¤§ W lh ¤¤¤¤*’¤l P•’°¤¤d'¤'¤ ¢¤V¢l°P¤d l'#Y74¤¤l. MJ _ -
‘ crysllllized from ethyl at:etate·petroleum ether. An ad- c°u¢W¤¤_C!$¢ll¤$l¤V· Ch¢¤t.C¤¤1¤t¤¤S.30. 1853 (19652. ’ ‘
ditional batch of product is obtained by evaporation gf R¤¤¤tS¢¤ll¤.¤¤Q¤ of the crude reaction product {rom dr-
un meme; muon and recryttallization of un insane. ¤¤=¤¤·lf¤r¤¤q¤d¤·w¤¢¢s vxelos 11-3 e. 06%) at ¤-t==¤zy§- ‘ -
Qver-all yield: ls.: g. (93%) ol pbenzylnrercaptonro- ¤¤irr;1¤¤¤pr¤l¤g¤¤§l.I’:1l;¤¤=y1-·L-gh=¤ylala¤yl·L·et¤¤ms-
P|0!l}:]·L·}}'l'¤II¤¤ methyl ester, M.P._89—90 C.; onucal 55 ¤§3· ¤¥1:¤'¤li¤¤¥_ · ZY§·L·¤Y5l¢¤-L1: ¤l¤\!¤Y| Hist'. M-?.
rotation: [»]n¤—3.4:.·0.5 (e. 0.5, dunethyunnnamldn). 2_ -255 Cnopqcal rotation: teln -29.6::0.5 {e. 0:, ·
For Cq.·.H¤N0.S (aus) orientated: 64.sr% c., 6.21% dnmethvlformamtdel. For C..HnN10..S,·V:H,0 t96S.tl
ra. 1.1s% Ngfonnd: susan c, 6.2:% H, :.11% N. ;¤l;¤l¤¢¢¤= 59%% G. 610% H. 10-27% N; !o¤¤d=
plknzylmercaptopropionyl-1.-tyrosyl-L-phenylalanine 9' 1% c' 633% H' 1023% N'
msthyl ester so Mtenzylmercaptopropiooyl · - tyrosyl-Lphenylalanyl·
B -Benzylmer¢aptopropionyl-L- tyrosine methyl ester Q" Elmamhyx ' L ' “P°"’“‘Y"s‘h‘“’Yl’L'°Y""”° bl"
Q5.! 3.3 ts_ ist ethanol gg} url.), solution ””d°
rstreate_wx 4 yrazine y te 6m.,a¤dthe A 1- { _h I - LL, up __
product is retluxed for 2 hours. An_ excess of water is 65 phcn5°.u21&lgyg J; fn:{,g:;$E3:g;gé¤y; ;, ' '
$*¤=¤ ¤¢¤;=¤ to the =l¤r¤r. the ¤¤¤¤¤¤¤ ¤= l=¤¤¤¤=¤ by =l>¤1v· ueyneine methyl ester uc.: nl in dirnetltylforrnamide
’¤¤ ’°°”°¤· *¤° ****9 ”’¤***=¤ “'*=¤= ¤¤ ¤== FM with use ml.) is treated wm. son ayunune hydrate uz nit.;
wm. and r=¤rv=¢¤¤¤¤·¤¢ fr¤¤= · - 11.95 e. (78%} of a-benzylm=r¤¤r{t¤vr¤1¤¤¤¤>·l·L·t¥r¤=i¤e mnnemnre ren 12 nears. The product which separates
hYdm‘d°· Ml 193*94. CJ °P'·‘“‘l ’°mi°”i 70 out is isolated by applying suction. washed on the tiiler
[¤]Dis_6_0:°_5· with methanol and ether, dried. Yield, 5.7 g. (6295) ·
_ of p - henzylmercaproproplonyl-Ltyrosyl-L·phenylala¤yl·
(e. 0.5. dlmethylformamide). For Cul-l¤N,0,S (373.5) L·glutaminyl-L-aspara:iny|·S-be¤zyl·L·¤YSl¤i¤¢ hvdnzlde.
calculated: 51.10% C. 6.2l% H. 11.25% N; Iound: M.P. 2664683 unchanged on recrystalliution [mm dl- ·
61.10% C. 6.11% H, 11.15% N. :5 methyl£ormamide·waler; optical rotalioot t·ln" -33.4

Case 1 :04-cv—00884-SLR Document 71 -22 Filed 07/17/2006 Page 4 of 4
E 3 S
3,497,491 _
5 6
z0.5' (c. 0.5, dimclbyliocmnmidc). For C;;H"N,0,S, purified by known procedure using a cnrtienfree bidi-
(956.1) calculated: 59.04% C, 6.01% H. 1319% N: voltage electrophoresis, el. Zaonl, M.; Sem; F.; collec-
f¤¤¤d= 59-04% C. 6-15% H. Q3-75% N- gachcdov;. £.dhempot:mD;a?._31, 310 (1966). Yield.
nz;.o eamn¤¤~ · ninine-vuonxesainguopti `
pamylmnupmpropionyl · I. - tyr¤syl·L·phenylala¤yI· 5 cal mtatlua [e1¤¤ -65:2' (c. 0.2, l M CH,000H). .
_ L-;lutaminyl·I.·aspara;Inyl·S ·l>euzyl·L-cy:teInyl·L· For C“H“N“D,,S,·2CH,00¤H (uv.:) calculated:
‘ "°"""°*°""°*'“‘“"""‘“"°"° .€.°·2§’f.§é§a‘°4’3.% L2s1‘.’§.§"*°"?."§.2¥2.F"%‘·3§»"£“
. . o ysrs: yr .
The compound stated ih this heading is prepared hom Plie 1.03,Gln 1.05,Asp 1.00. Pro l.0@,Arg 0.98: Gly 0.95. _ '
pbenzylmeruplnpmpinuyl - L - tyrosyl-],phe¤yhlmyl- lh What I: clalmcd as new and dented to be protected
Lglutamlnyl · L · uparaginyl-S-bcgzyl-L—cyatcinc hymn- by Letter! Patent is set forth in the appended claim:
zida and L·pxolyl·N°-t¤syl·D·u-ginybglycinamide in a 1. The polypeptide of the formula
68% yrcld acourdin to the pnexal procedure (vida
supra). M.P. 223·6' C. For C,-,I·I“l¤,.0,,S, (1405.65) §1•o-1·’P;r·!·P:•-1»¤}¤-I-A:=·¤>I»!»P;¤-D·A¥t-¤gsN¤•
‘ calculated: 51.24% C, 6.0295 H, 13.9595 N; tcuml: 15 _ _ _ _ _
$733% Q 60]* gt 13 sg; H_ whetcm Mop Inrltonlu pmcruphprcpmmc aud. _
mra¤1¤¤.a-¤.¤;s¤t¤¤-vadpreran ¤·f¤•¤¢¤ ¤¤·¤
,1 B lmeveapto mpio l l. tyxo lbph ylala 20 Im HATE PA S
— · wi! P ¤lv · - it · =¤ ·
;3*·*;;;_¤;:;g*,;,}_,;e·3;s;,·,*;,§,;,§d,°=·;:,*j;¤·;=·i;*¤;;; iziéiiil ‘$5}32§ £2§i.‘3.‘1':3Z{§;::;:§2?.:§3§
du};. with mdfm h'{iq{M mmm uwrlgl w we :,:11,0so 2/l96| ¤¤arr¤¤¤ama1. .... zsu-uz.:
gum! pmecdgrgnflvizlhe sw). ’1d'l;e{iq\:£tdi¤ax;tt;i’o¤k is 25 OTHER. REFERENCES
V en evaporate e r ' ua no ve aque- · _
....1. .... tm. ....... 1... ....¤.. .. mm .. *;:;*..3.%;t.”§;,.°,§*::$.’2?·£.¥.’;1§§%},""’
remove the uodissolvcd portion and the tiltrzlc is adjusted hm.;] tt al, é°“_ Cach Cbém .c°mmun_ ji 3g2__
to a pH of 6.5 to 'I by addition ol aqueous sodium by- 3;, u965)_ °
"..‘.°.....*"‘° *1.** .2% “}°" 3*1.**% *3 ";‘.......°"" "‘.‘l°:$“a“ .·»
3 ' YWE . ·» ra » · ·= · = ¤¤· LEWISGOTTS, Primary Examiner `
M Y.; d D vlgae ud, V.; J. Biol. Ch . 238, uu , ,
` ; (1963).·;'hc iluodunamiatum is lhcrcuptstfrnadittated to M· KASSEEOFE *`“"“"' E“m‘”"
' apI·l¤l4to4.5hyaddi1i¤n¤faceticac3d.‘l'hcpeptidc Us cl xx ’
isappliedtoaculumn ofzcaxbcxylate ion exchange " ‘ ‘ ' ‘
resin, it eluted with 50% aqueous cutie acid and isolated 35 260-470. $59; 424-—l77 ‘ .
by lyophillzatlon (tteeze-drying). He crude product is