CA2042468A1 - Compositions of and treatment with biologically active peptides having d-amino acid residues - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE

A biologically active peptide wherein each amino acid residue of the peptide is a D-amino acid residue or a glycine residue. Examples of such peptides wherein each amino acid residue is a D-amino acid residue or a glycine residue include deletion and substitution analogues of magainin peptides, CPF
peptides, PGLa peptide, XPF peptide, and derivatives of cholecystokinin. Such peptides have increased resistance to proteolytic enzymes while retaining biological activity.

Description

C (~ . .,r l D ~ 8 2 ~, G~&
~//~/.,~0 COMPOSITIONS O~ ~N3 TR~AT~E~T WITH
~IOLOGIC~LLY ACTIYE 2~PTID~5 H~IN~
: D-A~I~O ~CID ~SIDU~S
.
Th~s invention relate~ to biolo~ically ac~ive peptides, and more particularly to blolo~ically actiYe p~ptides wherein each amino acid residue of ~uch peptid~s i~ a D-a~ino acit residue or Ycine~
In accordance with an a~pect oP the present invention, there is provldod a compound &omprisln~ a~ analo~ue of Ma~ainin I
Peptide or Ma8ainin II peptide. ~ach ~mino acid resitue o~ the Magainin I peptid~ and the M~gainin II peptite is a D-amino acit residue or ~lyclne. Ths Ma~a~ni~ I o~ Ma8ainin II peptide i~ in an amid~ or carbo~y t~rminat~d form. Magainln I is repre3en~ed by thc follow~ng structural formuls u~in~ tha ~in~le letter amino acid code and the numbers b~low ~sch amino acid re~idu~ refer to th~ position o~ ~he ~esitua in th~ pep~id~
G I G R F L H S ~ a ~ F G K ~ F V ~ E I M ~ S
1 2: 3.4 5-~ 7~8 9 10 lI 12 13 14 15 16 17 18 l9 20 2~ 22 23 MaBainln II 1A r~preqented ~y ~he follsw~ng ~tructural ~o~mula u3ing tho si~gl~ Iettcr amino acit cod~ a~d th~ numbers below each a~ino acid residu~ refer ~o ~ho po~ on of the re~idu~ in th~ p~ptid~
G I G K F ~ H ~ A ~ ~ F G R ~ P V a ~: I M N S

' , - ' ~ ' ` ` ` ' -` '` . C ~ 2~ D ~ `.3, ~ 3 ~
The Magainin I orC~againin II peptide ls substig~uted ~ L~ ~2~
one of poYition3 1-23. The ~ubstituents which may be employed in esch of positions 1-23 are shown in the following table:.
Re~idue No. Substituent l D-Lys,D-Ala 2 D-Lys,D-Ala, D~ ,D-Ar~,D-Leu,D-Val,D-His,D-Met 3 D-Ly3,D-Al~,D-Trp,D-Ar~,D-His 4 D-Ly~,D-Al~,D-Ar6,D-Hl~
S D-P~Q,D-Ala,D-I.y8,D-Trp,D-Leu,D-Ile,D-Val,D-Met 6 D-Leu,D-Lys,~-Ala,D~ ,D-Vsl,D-Met 7 D^Ly~,D-Hls,D-Ala,D-Arg,D-Il~,D-Val,D-Met 8 D-Ala,D-Lys,D-Ser,D-Trp,D-M2t,D-Ile,D-Ar~,D-Hi~, D-Thr,D-Leu,D-Yal 9 D-Ly3,D-Al`a,D-Trp,D Arg,D-Hls,D-L~u,D-Ile,D-Val D-Lys D-Ala,D-Trp,D-Arg,D-Hi~,D-Leu,D-Il@,D-Val 11 D-Ly~,D-Arg,D-His 12 D-Phe,D-Ly3,D-Trp,D-Arg,D-His 13 D-Ala,D-Lys,D-Trp,D-M~t,D-Arg,D-His,D-Phe,D-Leu, D-Il2,D-~al 14 D-~la,D-Lys,D-Ar~,D-~i~
lS D-Ly~,D-Trp,D-Ala,D-~rg,D-His,D-Phe 16 D-Ala,D-Lys,D~Ph~,D-Ilo,D-V~l,D-Met,D-Leu 17 D-Ala,D-Ly~,D-Val,D-Trp,D-~rggD-~i~,D-Met,D-Leu 18 D-Ala,D-Lys,D-Trp,D-~rg,D-His,D-Leu,D-Met 19 D-~13,D-Lys,D-Glu,Gly,D-~r~,D-His,D-Leu D~ ,D-Phe,D-A3n D-Il~,D-Ala,D-~y~,D-Trp,D-L~u,D-Ph~,D-Val,D-Met 21 D-Ly~,D-Pro,D-Als,D-His,D~L~u,D-Arg,D-IlQ,D-Ph~
22 D-Ly3,D-Ala,D A~n,D-Gln,D-Arg,D-Hls 23 D-S~r,D-Lys,D-~la,D-Thr,Gly,D-Leu,D-Ile D-Gln,D-A~n ' r ~ F N T ~

Preferably, ~h~ ~bstituents are ~s f~ll-o.w~

Residue No. Substituent 1- D-Lys,D-Ala 2 D-Ly~,D-Ala,D-Ile 3 D-Lys,D-Ala,D-Trp 4 D-Ly3,D-Ala D-Pho,D-~la,D-Lys!D-Trp 6 D-L~u,D-Lys,D-Ala 7 D-Lys,D-Hi3,D-Ala 8 D-Ala,D-Lys,D-Ser,D-Trp 9 ~-Lys,D-Ala,D-Trp D-Lys D-~la,D-Trp 11 D-Lys 12 D-Ph~,D-Ly~,D-Trp 13 D-Ala,D-Lys,D-Trp 14 D-Ala,D-Lys D-Lys,D-Trp,D-Ala 16 D-Ala,D-Ly~,D-Phe 17 D-Ala,D-Ly~,D-Val,D-Trp 18 D-Ala,D-Lys,D-Trp 19 D-Ala,D-Lys,D-Glu D-Ile,D-Ala,D-Lys9D-Trp 21 D-Lys,D-Pro,D-~la 22 D-Lya,D-Al~,D-Asn 23 D-S~r,D-Ly~,D-Ala It i~ to b~ undorstood tha~ for purpo~e~ of th~ present invention, that th~ D-~ryptophan rasidu~ m~y bR protected wi~h a formyl gro~p or be unpro~ct~d. Tho ~-phenylalanlno re~idues, wheth~r ~uch r~sidu~ is pre~nt in lt~ normal poaition, or employ~d as a ~ub~tltution re~idu~, may bQ a normal D-phenylalan~n~ rQ~idu~ or an iodinat~d D-ph~nylal~nine re~idue.

-.: - ~

.
' ' . , C (~ r ~ T I ~ 2 ~ !~ 2 ~ ~ 8 In accordàn;~ë'wieh one embadiment.,-the peptld~ i'3 a Magainin I peptide~ and at lea~t one of smino acid residues 8, 10, 13, 16, 18 and 19 is sub~tituted with a D-~lanine r~sidue. In another embodiment, the Magainin I peptite has D-alanine residue ~ubstitution~ at each of sminQ scid re~idue3 3, 8, 16, 19 and 23.
In accordanoe with another embodiment, the Ma8sinin I
peptide has D-alanine r~idue substltutions at four of amino acid residues 3, 8, 16, 19 and 23, and the remalning one of these re9idue~ ls sub9tltuSed with a D-ly~n~ r~idu~.
In accortance with yet a further embodiment, amino acid 21 of a Magainin I peptide i9 substituted with a D-proline residue.
In accordance with another embodiment, at lea~t one of amino acid residu~s 3, 7, 8, 10, 18-21, and 23 of Magainin I i~
substituted wlth a D-ly~in~ residu~.
In accordance with anoth~r embodi~ent, amino acid re~idue~
3, 8, 9, 19 and 23 of Ma8ainin I a~a ~ach substltuted with a D-lysine residu~ and amino acld residu~ 15 i~ ~ubstituted with a D-alanine re~itu~.
In sccordance with a further embodiment, at least one of amino acid residue~ 3, 5, 8, 10, 12, 13, 15, 18, and 20 of MaKainin I is sub~tituted with 8 protect2d D-tryptophan residue, wherein the protecting group is prefera~ly a ~ormyl group. In accordance with anothor embodim~nt, at laast one of amino acid residues 9, 13, 15 and 17 of Magainin I is substituted with an unprotected D-tryptophan re~ldu~.
In accordancQ with anoth~r embodiment, the peptide ls a Magainin II poptld~ , and at least ono of a~ino acid re~idues 1- 8, 10, 13, 14, 16 as3d la-23 is a D-alanine re~idut!.
In accordan~ wlth ano~h~r embodlment, at le89t OII~ of amino a,cid residuQ~ 1-3, 5-9, 12, 13 snd 15-23 o Magainin II ls a D- lyslne re~idu~ .
Xn accordanca wlth anoth~r asp~c~ o~E th~ present invention, there i~ provided an an~logu~ of Ma~3~inin I or Magainln II
peptide, ~ald Ma~ainin I or Magainin II pep~id~ being in an .~

f~F~ ,T~ f2~7 ~
amide- or carbo~,y-t naeed form sn~ having the stru~tural ~
formulas hereinabove described, where.ln each amino acid residue of ~aid Magslnin I or Msgsinin II peptide i3 a D-smino acid residue or glycine, and wherein st lesst one o amino acid residue~ 15-23 is omitted and st lea~t one of the remaining amino acid re~idues is substituted. Th~ substituent~ which may be employed in st lesst one of positlons 1-23 sre shown in the following table:

' ' .
' ' .

C (? ~ i T IRe~idue No. ~ . Sub~tituent 1 D-Lys,D-~ls ~.~.: ; ;. . ..
2 D-Ly~D-Als, D-Ile,D-Arg,D-Leu,D-Val,D-His,D-Met 3 D-Lys,D-Ala,D-Trp,D-Arg,D-His 4 D-Lys,D-Ala,D-Arg,D-His D-Phc,D-Ala,D-Ly3,D-T~p,D-Leu,D-Ile,D-Val,D-Met 6 D-Leu,D-L~s,D-Al~,D-Ile,D-Val,D-Met 7 D-Lys,D-His,D-Ala,D-Arg,D-Il~,D-Val,D-Met 8 D-Als,D-Ly9,D-Ser,D-Trp,D-Met,D-Ile,D-Arg, D-Hi~,D-Thr,D-L~u,D-Val 9 D-Ly~,D-Ala,D-Trp;D-Arg,D-His9D-Leu,D-Il~,D-Val D-Lys, D-~la,D-Trp,D-Ar~,D-Hi~,D-L~u,D-Il~,D-Val ll D-Ly~,D-Arg,D-Hi~
12 D-Phe,D-Ly~,D-Trp,D-Ar~,D-Hi~
13 D-Ala,D-Ly~,D-Trp,D-M~t,D-Ar~,D-His,D-Phe, D-Leu,D-Il~,D-Vsl 14 D-Ala,D-Lys,D-Arg,D-Hi~
D-Ly~,D-Trp,D-Ala,D-Ar~,D-His5D-Phe 16 D-Ala,D-Ly~,D-Phe,D-Ile,D-Val,D-Met,D-Leu 17 D-Ala,D-Ly~,D-Val,D-Trp,D-Arg,D-His,D-Met~D-Leu 18 D-~la,D-Ly~,D-Trp,D-Arg,D-Hi~,D-Leu,D-Met,D-Phe l9 D ~la,D-Ly~,D-Glu,Gly,D-~rg,D-Hi~,D-Leu, D-Ile,D-Phe,D-~s~
D-Ile,D-Ala,D-Lys,D-Txp,D-Leu,D~Phe,D-Ysl,D-Met 21 D-Ly~,D-Pro,D-Ala,D-His,D~Leu9D-Arg,~ ,D-Ph 22 D-Lys,D-~l~,D-~n,D-Gln,D-Ar~,D-His 23 D-52r,D-Lys,D-~la,~-Thr,Gly,D-Leu,D-Ile, D-Gln,D-Asn , , r~ A~ 2~8 Preferably, the s~ituents are 83 follow~
Residue No. ~ Sub~tituent . .
1 D-Lys,D-Ala 2 D-Lys,D-Ala,D-Ile 3 D-Lys,D-Ala,D-Trp 4 D-Lys,D-Ala D-Phe,D-Ala,D-Ly~,D-Trp 6 D-Leu,D-Lys,D-Ala 7 D-Lys,D-His,D-Ala 8 D-Ala,D-Ly~,D-Ser,D~Trp 9 D-Lys,D-Ala,D-Trp D-Ly~,D-Ala,D-Trp 11 D-Lys 12 D-Phe,D-Lys,D-Trp 13 D-~la,D-Ly~,D-Trp 14 D-Ala,D-Lys D-Ly~,D-Trp,D-Ala 16 D-Als,D-Lys,D-Ph~
17 D-Ala,D-Ly~,D-Val,D-Trp 18 D-Ala,D-Ly~,D-Trp 19 D-Ala,D-Lys,D-Glu .D-Il~,D-Ala,D-Lys,D-Trp 21 D-Lys,D-Pro,D-Ala 22 D-Ly~,D-Ala,D-~n 23 D S~rtD-LyY,D-~la In accord~n~ with a particular embodiment, smino acid re3idu~ 16-23 o~ Magainin I ar~ d~ et, a~d amino acid residues 3, 8 and 10 ar~ ~ub~tituted with a D-a`lan~ne re3itue.
In accordanc~ wlth yQt anothsr embodi~nt, amino acid residue 19 of Ma~ainin I i~ deleted,~and pr~orably a~ino acid residue3 5~ 8, 9 a~d 16 are each sub~tltuted w~h a D-1ysine r~situe, amino acid re~id~ 21 i~ ~ub~t~tuted with a D-leucine .

~, , '' ' 6~ ,fr~ ~ ~ r ~ f~
re~idue, ant acid residues 18 a~d-2.3:ar~ sub~tituted with a D-alanine r~sidue.
In accordance with another embodiment, in carboxy- or amide-terminat~d (preferably amide-termlnated) Msgainin I or Magainin II peptide, amino acid re~idues 17-23 or 16-23 or 15-23 are deleted, and 8mino acid residues 3, 7, and 8 are each substituted with a D-lysinc residu~, and optionally amino acid residue 13 and/or amino acld residuQ 10 is sub3tituted with a D-alanine re~idue. Preferred peptides are as follows:

GIKKFLKKAGKFGKAF-NH~

GI~KFLKXAKKFARA-NH2 Preliminary ~tudies indicate thdt th~ abo~e-mentioned preferred peptides pos~ess low hemolyt~c act~vity. In accord~nce with a further embodimQnt, amino acid residu~ 21 of Magainin I may be del~ted, and preferably amino acid resldue~ 5,10, 18, and 19 are each substituted with a D-lysine resldu~, smino acid residue 7 i9 sub~tituted with a D-phenylalanin~ re3idue, and amino acid residue 22 i8 substituted wlth ~ D-alanlno re~$due.
In accordanc~ with ~noth~r embodlment, ln carboxy or smide terminated ~preferably amide ~erminated) Magainin I or Magainin II tPrefersblY Ma8ainin II), amino acid residue 19 i~ omitted, and st least ono o~ amino acid r~idu2s 3, 7, B, 10, 13, 15, 16, 18 21, 22 or 23 ia substituted with anothe~ amino acid ~g ~ollow~:
!i~ll!~ Substituen~
3 D-L~u 7 D-Ly~
8 D-Lys, D-~la D-Ala, D-Lys 13 D-Trp, D-Leu, D-Ph~, D~Aia D-Ph~
., .

lB D-Lys, D-Ala, DrPh;~, ~; . .
21 D-Lys, D-Ile - r!~
22 D-Ly~
23 D-Ly3, D-Ser In a preferred embodiment, the peptide is a Magainin II
peptide, a~d the substitution analogue wherein amino acid 19 is deleted is sel~cted from the claa~ con~istin~ o~ the following ~ubstitution analo~ues:
GIGKFLHSAXXFGRAFVGIMKS;
GIGXFLHSAKKFGRAF~AIMXS;
GXGKFLHSA~FFR~FVFIMNS;
GIGRFLKSARRFGKAFVFIMNS;
GIGXFLHKAKKFAKAFVFIMNS;
GIG~FLKSAKKFAKAF~FIMNS;
GIGKFLHRAKRFARAFVFI~
GIGKFLX~AKKFGKAFYFIMXK; and GIGKFLHSAXXF9RAFVK**IMNS; wherein K** i9~- F ~oc-ly~in~ w~ 0/~
Prellminary studiea indicato that the~o pr~erred analogues possess low hemolytic actl~ity.
In accordance with another 2~bodiment, ther~ are provided derivatives of Magainin I or Mag&lnln II (carbo~y- or :
a~ide-ter~inated), pr~erably Ma6ainin II, whQrein a portion of th~ basic peptit~ i8 telated and at lea~t one of the remaining amino acit re~iduo~ i~ subst~tutQt a~ h~reiQabo~e de~cribed; in partisular, ~m~no acid re~idu~ 19 i8 omltted and in addition : eithar a~ino 8cid ra9idues 1-4 or 3,5, and 6 sre omitted.
Pref~rred p~ptid~ ar~ as follow~:
FLHSAKR~GK~FYFIMNS
GIXHSAKKFAKAF~IMNS
FLRSAKKFGR~FVGIM~S
FLKSAgKF~XAFVGI~NS

,i - :

", C O ~ 13~ T ~` 1 2~
In accd~j,ance,~ith another aspec~.~of.the presant i~vention, there i~ provided a compound comprislng ~ pepti'de deletion analogue of an amide or carboxy terminated Magsinin I, wherein Magainin I i9 represented by the following structural formula using the sin~le letter amino acld code and the numbers below each amino acid residue refer to the pO9~ tion of the residue in the peptide:
G I G K F L H S A G K F G K A F V G E I M K S
1 2 3 4 5 6 7 8 9 10 11 1~ 13 14 15 16 17 L8 19 20 21 22 23 and wherein each amino acid re~idue 19 a D-smino acld residue or a gly~ine resldue; and at lea~t one of sm~no acid residues 15 through 23 is omitted. I~ a pre~rrQd embodiment, at least one of amino acid re~idues 1~, 16, 18, 19, 21, 22 and 23 i9 omitted.
In one embodim~n~, at lea~t a~ino acid residu~ 18 i~ omitted whereas in another embodiment, at lea~t amino acit residue 19 is omittad, and in yet anoth~r ambodimant ~t l~t ~ino acid residue 2$ i~ o~itted. In preferr~d embodlments, only one of amino acid residue3 18, 19, and 21 t re9p~ctively, i~ omitted. In other preferred embodiments, amino ac~d re~it~es 21, 22, and 23 are omittad, amino acid residue~ 19 through 23 are omitted, amino acid residues 18 through 23 are omittet, and amino acid residue~
17 throu~h 23 are omitted. The compound can be a deletion analo6ue of amid~ - or carbo~y-t~rminated Magainin I.
In accordance wlth a~other asp~ct of the pre~ent invention, there is provided a compound comprl~ln~ a peptide that i8 a deletion analo~u~ o an smid~ - or carboxy-terminated Magainin II, wher~i~ M~88in~n II is repr~nted by the followin~
structural fornula UBing the slngl~ letter amino acit code and the nu~bsr~ b~low ~ach amino acid r~ldu~ s~ar to ~h~ posltion o~ th~ rQsitu~ in th~ peptldQ:
G I G K F L H S A K K F G K A F V G ~ I M N S
1 2 3 4 5 6 7 ~ 9 10 11 12 13 14 lS 16 17 1~ 19 20 21 22 23, wherein each amino scid residu~ is a D-s~ino acid residua or glycine, ant at l~a~t on~ of amino ac~d~ 15 through 22 ls l .

- ; C ~ ~ 3 ~ L ~ ~ ~ g `_ . L ,, , ~, omitted. In a pr'eerred,emb,odiment, at lea~t,,one of ami'no a~ids 15, 18, 19, 20, 21, and 22 i~ omitted. In one embodim~nt, at lea~t amino acid 18 i9 omitted. In another embodiment, at least amino acid 19 i9 omitted. Another embodiment omit~ at least amino acid 21, and yet othe~ embodiment omits at least ami~o acid 22. In preferred ~bodimenta, only ono of amino acids 18, 19, 21, and 22, re~pectively, is omitted. The compound can be a deletion analoguc o amide-terminated Magainin II.
In accordance with yet another aspect of the present inven~ion, there i9 provided a baslc tpositi~ely charg~d) polypeptide having at lea~t ~ixteen amino aclds wherein the polypeptide includes at least eight hydrophobic amino acids and at least eight hydrophilic amino acid~. Each of the amino acid re~itues of the polypeptid~ i8 a D-amino acid resldue or a glycin~ re~iduQ. Still more partlcularly, the hydroph~bi~ amino acids are in groups of two ad~acent ~mino acids wherein the amino acid~ are D-amino ac1ds or glycine, and each group of two hydrophobic amino acids i~ spac~d from anoth~r group of two hydrophobic amino aclds by at l~a~t on~ D-amlno acid other than a hydrophobic amino acld (pr~ferably at least two D-amino acids) and generally by no greater than four D-amino acids, and the D-amino acids betwe~n pairs of hydrophobic amino acids may or may not be hydrophilic.
Th~ hydrophilic ami~o acids ar~ generally al~o in group~ of two ad~acent D-amino acids in which at lea~ one of the two D-amino acid~ i~ a ba~ic hydrophiliG a~ino scid, with such groups of ~wo ~yd~ophilic D-amino scld~ bein8 ~pac~d from each other by at least on~ ~ino acid, other than a hydrophillc D-amino acid, wherein 2~ch o~ ~ald at lea~t on~ amino acld(s) is a D-amino acid or glycin~ ~pr~f~rably at lea~t two smino acid~3 and g n rally no greater than four amino acid~, and tho ~mino acid~ b~tween pairs of hytrophili~ D-a~ino acids ~ay or may not ba hydrophobic.

~ .
,. ~ .
~: .

.

i ~ D
In accordanc~ with a particutarl~ pref~rred"embodiment,~the polypeptide compri~e~ a chain of st les~t ~our group~ of'amino acidq wher~in each amino acid i9 a D-amino acid or glycin~, with each group consisting of four amino acids wherein each ~f the at least four amino acids i9 a D-amino acld or glycine. Two of the four amino acids in each group are hydrophobic amino acids, and two of the four amino acids in esch gsoup are hydrophilic, with at lea~t one of the hydrophilic amino acids in each group being a basic hydrophilic amino acid and the other being a basic or neutral hydrophilic amino acld.
The hydrophobic amino acid~ may be seleoted from the class consisting of D-Ala, D7Cys, D-Phe, D-Ile, D-Leu, D-Met, D~Ysl, D-Trp, D-Tyr, snd Gly. The neutrQl hydrophilic D-smlno acid3 may be selected from th~ clas~ consisting of D-Ser, D-Asn, D-Gln, and D-Thr. Th~ basic hydrophilic D-amino aoids may be ~elected from the class consisting of D-Ly3, D-Ar~, and D-Hi~, snd D-ornithine.
Eaoh of the group~ of our amino scids may be of the sequenoe ABCD, BCDA, CDAB, or DA~C~ wher~in ~ snd B ara each hydrophobic amino acids and may be the sam~ or different, one of C or D is a ba~io hydrophilic amino acid, ant the oth~r of C or D
i~ a bssic or neutral hydrophilic amino acid ant may be the ssme or different. In a preferred embodlm~nt, the polypeptide chain may comprise 5 or 6 groups of this sQquenoe. In each group, each o~ A, ~, C and D ~ay be tha sa~ in ~om~ or all of the groups or may be di~fsrent in 30m~ or all of th~ groups.
Tho polypeptid~ chain preferably ha~ a~ lca~t 20 amino acids, and no ~roatQr than ~0 amino ~cids wh~rein each amino acid is a j-a~o acid or glycine. Xt is to be unders~ood, however, that thQ polyp~p~id~ does not hav~ to con~l~t entirely of the groups describet above. The polyp~p~lde may have amino acids extending from either or bo~h ends o~ thæ noted groups forming the polypeptido chain and/or thQr~ may b~ amino ~cits b2tw~en one or more of ~h~ at lea~ four group~ and ~tlll remain w~thin the , ~

~cope of th~ invention, provided that.each amino.s,c~d;-residue of the polypeptide chain-is a D'am~o acld residue or glycine.
The groups of amino acid~ may be repeatln~ groups of amino acids, or the amino acids in the various groups may vary pro~ided that in each gro~p of the at least four groups of amino acids there are two hydrophobic amino acids wherein each amino acid i9 a D-amino acid or glycine, and two hydrophilic D-amino acid~ as hereinabove noted.
Th~s, in 8 preferrad embodiment, th~ biolo~ically active polypeptide comprises a chai~ includlng at lea~t four groups of smino acids, each containing four amlno acids wherein each amino acid is a D-amino acid or glycina. Two of tha fous amino acids in each group are hydrophobic, wh~reln ~ach hydrophobic amino acid i~ a D-smino acid or glyclne, at lea~t ono ami~o acid i9 a basic hydrophilic D-amino acld, and tha remslning one is a basic or neutral hydrophilic D-smlno acid, wlth the polyp~p~lde chain preferably having at least 20 s~ino aclds but no greater.than ~0 amino acids.
In one embodlment, each of the ~t least four group~ of amino acit~ which ar~ in the p~ptido chain i~ of the sequence A-B-C-D, B-C-D-A, C-D-~-B or D-A-~-C wher~in A and ~ are hydrophobic, on~ o~ C or D i~ basic hydrophilic, and the other of C or D is ba~ic or neu~ral hydrophilic. The re~ulting polypeptite chain, th~refore, may hav~ on~ of th2 following seque~ce~:
(A-B-C-D)n(Y~)b ~ /30/~
/q~ (~21~~C~D~A)n(Y2)b (1~3)a(C~D~A~~)n(Y3)b (a~4)a(D-A-~-C)r,(Y~)b wherein ~1 18 D; C-D- or 8-C-D-, Y~ or -A-~ or -A-B-C

X is A-, D-A- or C-D-A-Y2 ~8 -B, -B-C or B-C-D

~3i~ B~, A-B- D~A-B~
Y3 is -C, -C-D, -C-D-A
~4i9 ~-, B-C-, A-B-C_ Y4 i3 -D, -D-A, -D-A-B
a iq o or l; b is o or 1 and n is at lea~t 4.
It i9 to be under~tood thAt th2 p~ptlde chaln may include D-amino acids or glycine between the hereinabove noted groups of four amino acids pro~ided thst the spacing between such ~roups and the ch~rge on the D-amino acidx or glycine does not change the characteristlc~ of the peptide chaln which provide amphiphilicity and a positive charge and do not adversely affect the foldin~ characteri3tics of the chain to that which i~
significantly different ~rom onc in whlch th~ hereinabove noted ~roup~ o~ four amino acid9 are ~ot ~paced ~rom çach other.
As reprosentative ~amplc~, th~re may bo mo~tloned.
I D-Ala-D-Phe-D-Sar-~-Ly~-D-Ala-D-Phe-D-Ser-D-Lys-D-Ala-D-Phe-D-Ser-D-Lys-D-Ala-D-Phe-D-Ser-D-Ly~-D-Ala-D-Ph~-D-Ser-D-I.ys II D-Ala-D-Phe-D-Ser-D-Ly~-D-Ala-D-Phe-D-Ser-D-Ly~-D-Ala-D-Phe-D-Ser-D-Lys-D-Ala-D-Ph~-D-Ser-D-Ly~-D-Ala-D-Phss-D-S~r-D-Lys-D-Ala-D-Phe-D-Ser-D-Ly~.
III D-Phe-D-Ser-D-Ly~-D-Ala-D-Ph~-D-Ser-D-Lys-D-Ala-D-PhQ-D-Ser-D-Ly8-D-Ala-D-Phe-D-Ser-D-Ly~-D-~la-IV D-S~s-D-Ly~-D-Ala-D-Phe-D-S~r-D-Lys-D-Ala-D-Pho-D-~r-D-Ly~-D-Ala-D-Ph~-D-Sar-D-Ly~-D-Ala-D-Ph~-D-Ser D-Lys-D-Ala D-Phe-V D-L~--Ala-D-Phe-D-Ser-D-Ly~-D-Al~-D-Phe-D-Ser-D-Ly~
D-~18~D-Ph~-D-S~r-D-Ly~-D-Al~-D-PhQ-D-Sor The peptid~ may hav~ amino acit~ e~tandin~ from either end of the chain~ For example, the chaln~ ~ay h~ a D-Ser-D-Ly~
sequenc~ ba~ore the "D-Ala" end, ant/or a D-~la-D-Phe ~equence after th2 "D-Ly~" snt. Other amino acid ~equences may al~o be attachod to the "D-Ala" and/or ~h~ "D-L~" end.
--.

CO~ 3 ~ ~2t~ j Similar~y, in.any poly~eptid~..chaln of.the.pr~se~t ~~
invention having atnlea~foù~rrgroups-o~ amino acid~;;of.ithe sequence as described above, the chain may hsve, for exsmple ~ a C-D sequence before the first A-B-C-D group. Also other acid sequences including D-amino acid~ or glycine may be attached to the "A" and/or the "D" end of one of these polypeptide chains.
Also there may be D-amino acids or glycine residues in thP chain which sp~ce one or more groups of the hereinabove noted four amino acids from esch other.
Such polypeptides ~re generally water-soluble to a concentration of at lea~t 20 mg/ml at n~utral pH in water. Such polypeptides are n~n-hemolytic, l.e., it will not rupture red blood cell~ at effective antimlcrobial conc~ntrations. The ~tructure of quch polypeptide~ provides for flexibility of the polypeptide molecule. When tha polypeptide is placed in water, it does not as6um~ sn amphiphilic structur~. When th~
polypeptide enco~nt~r~ an oily surfacQ or membrane, the polypeptid~ chain fold~ upon it~elf lnto a rod llke structure.
In accordanc~ wlth a furth~r 8~pact of tha present invention, there ls provited a peptide (polypeptidc) having from eight to fifteen amino aclds comprised of at least four hydrophobic amlno acids ant four hyd~ophilic amino scids. Each amino acid residu~ is a D-am~no ac~t re~idue or glycine. The hydrophobic amino acids are in groupY of two ad~acent hydrophobic D-amino acids or glycine wherein each group o~ two hydrophobic D-amino acids or ~17cinQ is spaced from each other by at least one D-amino acid other than a hydrophobio D-amino acid or glycine (preferably ~t l~a~t tw~ D-amino acids) and generally no greater than four D-amino acids, ant th~ D-amino acid(s) b~tween pairs of hydrophobic a~ino scgts wh~r~in e~ch am~no acid ia a D-amino acid or glycine9 may or may not bQ hydrophilic. The hytrophilic D-amino acid~ ara gen~rally alao ~ groupa of two ad3acent D-ami~o acid~ in which at lea3t one of th~ two n-amino acids i~ a basic hydrophilic D-amino acid snd th~ oth~r of th~ two i~ ba~ic .

or neutral. Tfi~group3 of two .D-hydrophilic ~min~,açid~ sre ~paced fro~:~ach othjer by.-at l.ë~ o~e'~ mino!.acid'.ot.~er than a hydrophilic D-amino acid, and wh~rein the at least one amino acid i9 a D-ami~o acid or glycin~ (preferably at le~st two amino acids) and ~enerally no greater than 4 ~mino acids, and the amino acids between pair~ o~ hydrophilic amino acid~ may or may not be ` hydrophoblc.
The peptide havin~ ~rom 8 to 15 amino acids is amphiphilic snd i~ po~ltively charged (basic).
The 8 to 15 amino scid peptldo hereinabove described may or may not be bioactive, and in the ca~e where such peptide is not bioactive, it hss utility a~ an lnterm~diate in provid$n~ the hereinsbove noted peptide~ which h~va at lea~t 16 amino acids and which are bioactive. For exampl~, two pQptid~s, one having eight D-amino acid3 and the other ha~ing twel~e D-amino acid~ may b~
coupled to each other to form a peptido havin~ 20 D-amino acids of the type hereinabove described and whtch i8 bioactive. The peptides may be coupled by standard peptite chemi~try technique~.
Thu3, for example, such peptides may b~ conden~et in solution by the technique disclosed by Johnson, ~t al. PePt~te~, pages 239-42 (Walter de Gruzter & Co., 19~6).
A3 repre~entative e~amples of such peptides, there may be mentionet peptid~Y rapresent~t by th~ following structure wherein A, ~, C and D are as defined prQ~iously;
(i) (Wl)a(A-E~~C~D)n(Zl)b (ii) (w2)a(B-c-D-~)n(~2)b (~li) (w3)~a(c-~-A^B)n(z3)b (IV) (w4)a(D-A-~-c3n(z4)b wherein Wl i~ D-, C-D-~ B-C-D-W ~ A-, D-~-, C-D-~-W is B- A-B-, D-A-B-W4 is G-, B-C-, A-B-C-Z i8 -A, -~-B, -~-B-C

:, ~

,~ ' , .

2~7~

Z2 i~.-Bj :B-~C, -9-C-D-. "~
Z3 lS~ D~ D-A~
Z4 is -D, -D-A, -D-A-B
n ig 2 or 3, a i9 o or 1 snd b i3 o or 1.
In accordance with yet another sspect of the present invention, there is pro~ided a peptide which include~ the following ba~ic peptide 9tructure ~:
-Rl-Rl-R2-R2-Rl-Rl-R3 ~1 -Rl-Rl-R3-Rl-Rl-R4 R~ ~1 whereln Rl is a hydrophobic amlno acid;
R2 i~ a hydrophobic amino acid or a b~sic hydrophilic amino acid;
R3 is ~ basic hydrophilic ~mlno acid;
R4 i8 a hydrophobic or n~utral hydrophilic amino acid;
R5 is a ba~ic or neutral hytrophillc amino acld, and each amino acid residu~ of said peptito i~ a D-amino acid ra~idue or glycine.
The her~inabov~ basic st~cturo i~ har~nafter ~ymboIically lndic~ted a8 ~. Paptide~ formed in 6ccortanc~ with this ba3ic ~tructure ar~ com~only re~erred to a~ CPF peptide3.
The CPF peptides of the preaant invention may includ~ only the hereinabov~ noted D-amino acids or glycine or may include additional D-amino ac~ds or ~lycino r~ldu~a at th~ amino and/or carbo~yl end or both th~ amino and carbo~yl end. ~n general, the peptide does not include more than 40 amino acid~, wherein each amino acid 1~ a D-a~ino acid or 61ycine.
Th~ CP~ poptid~s incl~ding th~ abov~ ba8ic 9tructur~
preferabl~ h~ rom 1 to 4 atditional amino acid~ at the smino end, wh~Ql~ ~ch amino acid is a D-ami~o acit or glyclne.
Accordingly, ~uch pr~f~rrQd paptida~ may b~ r~pse~ented by the structural ~os~ula:
y whereln ~ i8 thc hereinabov~ d~crlbet ba~lc peptide 3tructure ant Y 18 .

.

O ~ d~ 2 ,~

(ii) '~. ( R2-R5~.;,or-~
(iii) Ri-R2-R5; or (iv) R2-Rl-R2-R5i pre~erably Glycine - Rl-~2-R5.
wherein Rl, R2 and R5 are a~ previously dafined.
The carbo~yl end of the baslc peptida struoture may also have additional amino acits which m~y range from 1 to 13 additional amino acids.
In a preferr2d embodiment, tho basic structure msy have from 1 to 7 additionsl smino scid~ at the carboxyl end, which may be represented as follow~:
-X - Z whereln is the h~reinabove dQfined ba~lc peptide structure and-Z
i3 (i) Rl-, or (ii) ~1 Rl ; or (iii) ~l-Rl-R4; or (iv) Rl Rl R4 R4; r (v) R~ -R4 R4-R~; or (vi) R -Rl-R4-R~-R6-D-Gl~; or (vii) Rl-Rl-R4-R4-R6-D-~ln-D-Gln, wherein Rl and R4 are a~ previou~ly tefined, and R6 1~ D-prolin~ or a hydrophobic amino acid.
Pre~errad peptida3 may b~ repre~ont~d by the following structural ~o~ula (Y)~ - X ~ (~)b whæse~n ~, Y ant Z are a~ prQ~io~sly d~fined snd a is O or 1 and b 18 0 or 1.
A3 repre~ntativa e~ample~ of CPF peptld~ used ln the present inve~tion, th2re may b~ ~ntioned peptide~ represented by the followlng (~in~le l~tter anino ~clt code):
Gl2S3LG4ALKASLKIG678LCG9(10~QQ

.
, . " , ~ ., ~

. - ' ~

I

- C (~) N F ~ iT ~ A~ f5 ~ ~
bhere ~ s~ - , 2 - G, A
3 ~ F, L
4 ~ K, L

5 = A, G, T

6 = A, T

7 ~ H, N

8 = A, M, F, L

9 = A, S, T

10 = P, L
Preferred CPF peptide~ are of th~ following ~equences:
(1) GFGSFLGLALKAALKIG~NALG~APQQ
(2)~ GLASFLGKALRAGLKIGAHLLGGAPQQ
(3) GLASLLGX~L~AGLKIGTHFLGGAPQQ
(4) GLAsLLGKALKATL~lGTHFLGGApQQ
: (5) GFAS~LGXAlX~ALXIGANMLGBTPQQ
: (6) ~FGSFLGK~LKAALKIGANALÇGAPQQ
: (7~ ~FGSFLGRALKA~LKIGANALGGSPQQ
:: (8) GFASFLG ~ LKIGANLLGGTPQQ
In accordanc~ with an aspect of th~ pre~nt invention, there is provided a compo3ition compri~ing at les~t on~
biologically activc poptido whlch incIudes at l~a~t a chain`of th~ followin~ 20 a~ino acid r~3idu~ h~vin~ the following peptide structure ~10 wh~r2 ~lO i9 11 12 11 ll R13 R13 Rll Rll R12^~ R15 ~l4^Rll^R -R
: Rll R16 ~15 R17, wh~rein ~ : ~R~ a ~ydrophobic amino acid;
; Rlæ i~ :a ba~ic hydrophilic~amino ~cid; ~
:R13 l~ a~hydrophobic:~or:ba~lc~:hydrophilic amino acid; ~ :
Rl4 i8 a n~tural hydrophillc~a~ino~;-c~d,~or a ba~ic :
hyd~ophilic amlno acid;
R15 ls a neutral hydrophll~c a~ino acit; Rl~ is ~^proline or a hydrophobic a~o acit; and:R17 1~ a n~tural hydrophilic, .: : :
:: : ~ ' ;:

: ~ :

.
- ' : ' ' : : . . ~
.
, .

~ C O ~ ,. '30 , . ~ J~8 ba3ic hydrophilic or hydrophobic am~no acid. Each;of~th~ ~mino acid re~idu~a of the peptide i9 a D-amlno acid residue or glycine. Th~ at l~a~t one biologically active peptide which include~ peptide residus X10 has at la~st the hereinabove described twenty amino acid peptide and generally no greater than 35 amino acids, preferably no grc~ter than 30 ~mino acid~. Such peptides are derived from fra8ments of the human hormone cholecystokinin, or may be derivsti~e~ o such fragments.
In one embodiment, the at lea~t one blologically active peptide i9 of the fo~mula Rll-Rll-X-, wherein Rll i9 a hydrophobic amino Rcid as de~cribed abo~e. Mo~t preferably, the biolo~ically activ~ peptide of thls emhodiment include~ a chsin of at least th~ following amino acids:
D-Leucine-D-Leucine-X-.
In another embodiment, th~ at l~a9t on2 biologicslly active petpide includes a chain of at leaat the fcllowing amino acids:

12 11 15 Rll Rll , wher~ln Rll ' R12 ~ ~nd R15 are amino acids of the type3 hereinabove descrlb2d.
The peptidQ may be amide-t~rminated or carbo~y-terminated.
In ~ccordance with still anoth~r aspQct of the pre~ant invention, there ~ 8 provid~d a biolo~lc~lly ~ctiv~ ~mphiphilic peptide which include3 the following ba~lc structure X12:
~ R21 R22 R22 R~3~R21~R22~R22~n- whoreln R21 i~ a basic hydrophilic amino acid, R22 ia a hydrophobic amino acid, R23 is a neutral hydrophilic or hytrophobic amlno acid, n i3 from 2 to 5, and esch am~no acid re~iduQ ~f the pep~it~ i~ a D-amino acid residu~ o~ ~lycin~.
In ~ccostanco with one e~bodl~nt 3 the peptide may include th~ followln~ st~ucture:
Y12-~12, wh~r~in X~2 i9 a3 h~in~boY~ de~cribed, and Y i9:
(i) R22;
(ii) ~22-R22;
(ii~) R21-R22 R22;
(iv) R23-R2l-R22 ~22;
, .

CO~ 3 ~A~ 2~
tv) ~22-R23-R21-R22 R22;
t ) R22 R22 R23 R21 R22~R22~ whPrein R21, R22, and R23 are a~ h~reinabove de~cribed In accordance with another ~mbodiment, the peptide may include the following ~tructure:
X12-Zl2, wharein X12 is a~ herelnabove de~cribed, and Z12 i9 :
(i) ~21i (ii~ R21-R22;
(iii) R21-~22 R22;
(iv) Rzl-R22 R22 R23;
(~r) R21-R22-R22-R23 X21;
(vi) RZl-R22-R22-R23 R21 22 I~ accordance wlth y~t anothor ombodi~en~, the peptid~ may include thc followi~g ~tructure:
(Yl2)a ~12 (Z12)b, wherein Y and Z ~r~ a~ previously defined, a is 0 or l, and b i8 0 or 1.
lr~ a preferred embodiment, n i~ 3, and mo~t preferably the peptide i~ of th2 following structur~ as indicated by the single letter amino acid code:
~KIAGKIA]3~
In another embodiment, n i~ 2, and th~ peptide preferably i9 of the following ~tructur~ a8 indlca~et by th~ single letter amino acid cota:
KIA(K~AG~IA)2~IAG.
In accordanc~ *ith another a~pcct o~ the pre~ent i~v~ntion, ther~ 18 pro~id~d a blologic811y ~ctlv~ amphlphlllc p~p~ide which includ~s th~ ~ollowi~g basic structur~ ~14:
R21 R22 R22 R23 ~21 R22 ~22 R21 R22-~22-R22-R21-R22-X22' wh~rein R21, R22 and R23 are a~ h~rslnabov~ de~crlbed, and esch ~mino acid reoiduo o~ th~ p~ptid~ io a D-amino aclt r~3idue or gl~cin2.
In accordance with one ~mbott~nt 7 thQ peptld~ msy include the ~ollowin~ 9tructure:

C ~ L~
Y~ w~reln ~l4 i~ as-herelnabove describêd, and Y14 i9 :
(i) R22;
(ii) R22-R22;
(iii) R21 R22 R22;
(~v) R23-~21-R22-R22;
(v) R22 R23 R21 ~22 R22;
(~i) R22 R22-R23-R2l-R22-R22l or (vii) R21 R22 R22-R23-R2l-R22-R22~wherein R21, R22 and R23 are as hereinabov~ described.
In accordance with anoth~r embodiment, th~ peptide may include tha following structure:
X~ 4, wborein ~14 i~ a~ h~r~ns~ove d~scribed And Z14 is:
(i) R21;
(ii) R21-R22;
(iii) R21 R22 ~22;
(lv~ R2l-R22-R22-R23;
~v) R2.1-R22-R22-R23 a21;
~vi) R21 R22-R22-R23-R2l-R22; or (vii) R21 R22 ~22-R23-R~}-R22-R22, wh~r~in R21, R22 and R23 are as her~inabove de~crib~d.
In acco~tanca with yet anoth~r ~mbodiment th~ pept~de may include th~ following structuro:
(Y14)a-~14-(Z14)b, wh~rein ~ and Y ar~ a~ pre~iou~ly definet, 8 i~ O or 1, and b i8 0 or 1. In a preferred emboti~nt, th~ p~ptld~ is o~ th~ followin~ structural ~ormula a~
indicat~d by th~ Sin~lQ letter amino aoid cod~:
KLAS~AG~I~GKI~XV ~ .
In anoth~r prQf~rr~d ~mbodi~nt, tho p~ptid~ i5 of the following ~tructural formula 8S indlcat~d ~y th~ ~lngle l~ttes amino acid cod~:
KIAGKIA~IAGOI~RIA~KIA.

, , . , ., . :, .. .. .

,. ::

.

C (~ r ~ NT ~ V { ~ i2 ~ ~
In accordance with a further emb.odiment,.the peptide may include the followlng basic peptide structure X16:

R3l R34 R32 R31 R31 R32 R31 R3l R31 R32 wherein R31 i9 a hydrophobic amino acid, R32 is a basic hydrophilic smino acid, R33 i~ a neutral hydrophilic, b~sic hydrophilic, or hydrophobic amino acld, and R34 is a hydrophobic or basic hydrophilic ami~o scid, and ~ach amino acid re~itue i~ a D-amino acid resitu~ or glycine. Pr~ferably9 R33 i9 a neutral hydrophilic amino acid.
Such peptid~s ~re commonly reforred to as PGLa poptite~.
The PGLa peptides generally inclu~e at least seventeen amino acids and may include a~ ~any ~8 forty amino acids.
Accortingly, the hQreinabov~ describ2d ba~ic peptide structure for a PGLa peptide msy includ~ additionsl amino acids at the amino end or at th~ carboxyl ~nd or ~t both thQ amino and carboxyl end.
Thus, for example, 8 PGLa paptido may havQ thQ following structure:

wh~ra X16 is a pre~ioualy d~fined; and Y16 i8:
(i) R31; or (ii) R34-R31 wher~ R31 and R34 are as previously defined.
For Q~ampl~j a IGLa peptidQ may ~lso hav~ the following structuro:
~16 Z16 wh~r~ ~16 ia a pr~viou~ly d~ined; ant Z16 is:
Y16 i9 (i) R31; or (ii) Et'31-R31 where R31 is as previou31y de~ined.
A PGLa peptid~ may also have the following struotre:
, C O ~ 2 ~ 8 (Y16)a 1-~ ~ 16)b wh~r~ ~16~ Y16, and Z16 ar~ as previously defined, a i~ O
or 1 and b is O or 1.
In accordance with another embodiment, the peptide may include the following basic peptide struct~re X18:
R31 R33 R32 R31 R34 R35 ~33 R31 R31 R31 R32-(R36)~-R31--' wherein R31, R32, R33, and R34 are as previou~ly deflned, and R35 i~ glutamlne or a~paragine or a basic hydrophilic, or hydrophobic amino acid, R36 i3 glutamic acid, aspartic acid, a hydrophobic amino acid or a basic hydrophilic amind acid, and n is 0 or 1. Each amino acid residue is a D-amino acid re~idue or glycin~
Such peptides are commonly referred to a~ XPF peptides.
The XPF peptide~ generally includ~ at l~a~t nlneteen amino acids and may include up to forty amino a~id~. Accordingly, the hereinabove described basic peptite 3tructure oP XPF may include additional amino acids at the aminn and carboxyl end~.
Thus, for example, an XPF peptlte may includa the following ~tructure:

where ~18 is a~ prevlouQly dePinet ~nd YlB i3 (i) R31 or (ii) R34-~31 where R31 ant R34 are ~g prevlou~ly defined.
An gP~ pop~ido may include the followln~ structure:

whQr~ ~18 i8 a~ pr~io~sly d~1n~d nd Z18 i9 (i) R31; or (~i) R31-R35; or (iii) R31-R35-D-Prolin~; or (iv) R31-R3,~-D-Pr~ e R32 An ~PF peptide ~ay al#o hsve the following 3tructure;
~Yl8~8 ~18(Z18)b .j wh~r~ , Y18land Z18 ar~ a~ pre~;iously defin~dj a is 0 or 1, and b is 0 or 1.
Preferred are ~PF or PGLa peptides, which sre characterized by the following primary amino acid sequence (single letter amino acid code):
PGLa: GMASKAaAIAG~IARVALKAL (NH2) XPF: ~SKIGQT~KIARVGLKELIQP~
A review of XPF and PGLa can be ~ound in Hoffmann et al., EMBO J. 2:711-714, 1983; Andreu et al., J.~iochem, 149:531-535, 1985; Gibson et al. J. ~iol Chem. 261:5341-5349, 1986; and Giovannini et al, ~iochem J. 243:113-1~0, 1987.
Applicant~ unexpectedly have found that the above-mentioned peptid~, all of which consist entirely of D-a~ino acids or glycine resldue~, have biological activity.
The abo~e-mention~d peptide~ alao hsve increa~ed resi~tance to proteolytic enzymes while retsininB their biological activity.
In general, the peptide~ hereinabo~e described, and/or analogues or derivatives thereo~ are generally water soluble to a concentration of at least 20 m3/ml ~t neutral pH in water. In addition, such pcptides are non-hemolytlc; l.e., they will not ruptur~ blond cell~ at effactlvo c~ncontrations. The peptide~
cau~e less than 5X hemolysis of human erythrocytes at a concentration of 500 ~glml. In ~ddition, the structure of ~uch peptlde provides for fle~ibilitr of eh~ p~ptida molecule. When the peptide i3 placet in water, it does not s~sume an amphiphilic structure. When tho p~ptid~ encount~r~ an oily ~urface or membran~, thQ peptid~ chain ~old~ upon its~lf lnto a rod-like s'sructur~ .
Th~ peptldes m~y b~ C- tern~ln~l aclds or a~id~s .
The pept~d~s may be admini~tered to a ho~t; ~or example a human or non-hu~an animal, in an a~ount ffectiv~ to inhibit growth o~ a tar~t c~ll or ~irus. Thus, for e~mple, the peptides and/or analo~ues or derivativ~ theroo~ may b~ used as antimicrobial agen~s, anti-viral agent~; antibiotic~, anti-tumor :

:~

. ' :

,C~ ..2~F,N"T,l,~ ,J~t~
agPnts, sntlpar~iti;c agent,s, spermicide~.,.as-well a~ ~xhibiting other bioacti~e functions.
The term "antimicrobial" 88 used herein means that the peptides of the pre~ent invention lnhlblt, prevent, or destroy the growth or proliferation of microbes such a3 bacteria, fungi, or ~he like.
The term "~ntlbiutic" as used h~rein means that the peptides employed in the present inv~ntion produce effects adverse to the normal blological functlons of ~he cell, tissue, or organism includlng teath or d~structlon snd prevention of the growth or proliferation of the biological ~ystem when contacted with the peptide3.
The term "spermicidsl" 8~ u~ed herein means that the peptides employed ln the present in~ntlon, lnhibit, prevent, or d~stroy tha mot~ y of ~perm.
The term "antiYiral" as u~ed herein means that the peptides employed in the present inv~ntion inhibit, prevent, or destroy the growth or prollferatlon of virusa~.
The term anti-tumor a~ u~d hor~in means that the peptide inhibit~ the growth of or de~troy~ tumor~.
The term "antlparasit~c" a~ uset herein means that ~he peptides of the pr~s~nt ln~ntlon may be usQd to inhibit the growth of or destroy para~ita~.
Th~ pap~itos of ~ho pr~sQn~ inv~ntion hava a broad ran8e of potent antibiotic ac~ivity a6ainst 8 plurality of microorganisms includin~ Gr~m-posltivo and Gram-ne~atlve baoteria, fungi, protozoa, ~ h~ lik~, as well a~ parsslt~3. The pep~id2s of the pras~nt lnvontlon allow a m~thod for treatin8 or controlling microblal in~ction c~usQd by or~ani~m~ whlch aro ~n~itiv~ to the peptid~s. Such t~atm~n~ ~ay comprlse sdmlnistering to a host organism or ~lasu~ suaceptlbl~ to or af~lliat2d with a microbial lnfection an antimicrobial amount of at l~a~t one of th~ peptites.

!~ . . .

CO~2ir~ )E~`' ~ ' `'~ I
3 L~ 2 ~ ~ ~
B~cau~ of the intiblotlc propertles of th~ peptid~s, they may also be u~d as preservative~ or sterilant~ of materials 9US ceptible to microbial contamination.
The peptide may be administered in combination with a non-to~ic pharmaceutical carrier or vehlcle ~uch as a filler, non-to~ic buffer, or physiologicsl saline solution. Such pharmaceutical composition~ may be used topically or systemically and may be in any suitable form such a8 8 liquid, solid, semi-solid, in~ectable solu~ion, tablet, ointment, lotion, paste, capsule, or the like. The peptide compositions msy also be u~ed in combination with ad~uvants, protea~e i~hibitor~, or compatible drugs where such a combination is aeen to be desirable or advsnta~ous in co~trollin~ inaction cau~d by harmful microor~anisms including protozoa vlru~e~, and ~he like, ~ well as by parasite~.
The peptides of th~ pre~ent invention may be administered to a host; in particular an animal, in an effec~ive antibiotic and/or anti-tumor and/or anti-viral ant/or anti-microbisl and/or anti-para81tic and/or an anti~per~icidal amoun~.
Depentin~ on th~ u8e, A compo~iSlon ln accord~nce with the invention will contaln sn Qf~ectiva anti-microbial amount andlor an effective antisp~rmicidal amount and/or an effec~lve anti-viral amount and/or an effective anti-tumor amount and/or an effective antlbiotlc amount snd/or anti-parasitic a~ount of one or mora o~ the herelnabove described paptide~ which hsve.such activity.
Th~ pQptidQ of the present invention may al90 be employed in promotin3 or ~timulating healing of a wound in a host.
The ter~ "wound h~aling" a~ us~d herein lncludes various aspect~ of th~ would healin~ process.
These a~p2ct9 include 9 but are not llmited toS increa~ed contraction of thQ wound, increa~ed d~poaitlon of connactlve ti~3ue, a~ evitenced by, for e~s~ple, lncreas~d depo~ition of collagen in the wound, 8nd incr~ d t~n~lle s~rength of th~

C~ 8~NT'i,',' wound, i.Q. ,~:the peptides increase wou~drbrea~ing strength. The peptides of the present in~ention may alao be employed ~o a~ to rever~e th~ inhibition of wound healin~ cau~ad by steroids such a~ cortisone.
The peptides of the presant invention may bo used in the treatment of externsl burn~ and to treat snd/or prevent skin and burn infections. In particular, tho peptlde~ may be used to treat skin and burn infection9 cau~ed by or~anisms such as, but not limited to, P. aeruRinosa and S. aureus.
The p~ptides are also u~eful in the prevention or treatment of eye infections. Such inf~ction~ may be caused by bacteria such as, but not limited to, P. aeru~lnosa, S. aureu~, and N.
onorrhoeae 9 by fungi such a~ but not limited to C. albicans and A. fumi~atus, by parasit~ ch aa but not liml~d to A
ca~t~llani, or by ~iru~e9.
The peptidQs may al~o b~ eff~ctlve in killing cy9t8, ~pores, or trophozoites o infection - c~using or~anism~. Such organi~ms include, but are not limitcd to Acanth~moeba which forms trophozoit~ or cyst~, C. albicsn~, which form~ spore~, and A. fumiRatus, which forms spore~ Q~ w~
In general, the peptid~ ia employed to provide peptide dos~es of from 0.1 mg. to 5G0 m~. per kilogram of host weight.
When admini3tered topically, tha pQptide i~ u~ed in a concentration of ~ro~ .05% to 10%.
The peptide~ may be protuced by known techniques and obtainod in subatantlally pure form. For e~smple, the peptldes may bo syntho~iz~d on an auto~atic synthe~iz~r. Journal of the American ~h~lcal SocietY, Vol. 85, pg~. 2149-54 (1963). It i~
al~o pos~lbl~ to produce such pQptid~ by g-netic en8~neering techniques.
The peptidcs of the pr~s~nt in~ntion ~ay bo at~inistercd alona, or ln combination with oth~r ~uch paptide8. The peptites may al~o b~ ~d~iniatered in comblnstion wi~h biologically active peptidea or prot~ns con~aining L-amino acid re~iduo~, huch as ,.
:
~, :

C C~ ~ t ' , ~ 2 ~ ~ g ion channel-fo ~ ng protein~, or the p2ptides of:the presenc invention may be admini~tered in combinat~on with other active components, such as to~ic ions or antiblotics.
BiologicAlly active peptid~ containlng L-amino acid re~due~ which may be administer~d ln combination with the peptide~ o~ the preaent invention lnclude magainln peptides, PGLa peptides, XPF peptide~, cecropin~, and sarcotoxins. Magainin peptide~ ar~ described in Proc. N~tl. Ac~d. Sci., Vol. 84, pg9 544~-53 (AU8~ 1987). A review o~ ~PF ~nd PGLa peptides may be found in Hoffman, et al., EM~OJ., 2:711-714 (1983). Andreu, et al., J. Biochem, 149:531-S35 ~198S); Gib~on, et al., Biochem J., 243:113-120 (1987).
The cecropins and analogue~ and derivatives thereoP sre described in Ann. R~. Microbiol., Vol. 41, p~, 103-26 (1987), and Christensen, et al., PNAS, Vol. 85, pga. 5072-5076. ~1988).
Sarcoto~ins and anAlogue~ and tarivati~es thereof are tescribed in Molecular Entomotogy, pg5 369-7B, Alan R. Li~s, Inc. (1987).
Ion chann~l-fonmin~ protein~ or pQptidQs which may be employed in combination with ~he p~ptld~s of the present inYention includ~ dQfe~sin~, al80 known A~ human n~utrophil antimicrobial peptites (HNP), ma~or ba~ic protein (MBP) ~f eosinophil~, bactaricidal pormaability-increa~$n~ protein (BPI), ant a pore-forming cytoto~in cAllod varlously parforin, cytolysin, or pore-formin~ protoin. Do~n~ins are de~c~ibed in Selstet, et al., J._Clin. Inv~st., Vol. 76, pgs. 1436-1439 (198S). M9P protQins are d~cribed ln Was~oQn, et al., J. Biol.
Che~., Vol. 263, p~. 12559-12563 (1988). BPI protQin~ are describ~d in Ooi, ~t al., J. Biol. Chem. Vol. 262, pg8.
14891-14894 (198~). Perforin is d~cribsd in Henkart, et al., J.
Exp. Med., 160:75 ~1984), and ln Podack3 ~t al., J. EXP. Med., 160:695 (1984). Th~ abo~e articl~ ara h~reby ~ncosporated by reference.
To~lc ion~ which msy be e~ploy~d ln co~binstion with the peptid~3 o~ the prQsent inYen~ion includ~ anion~ 3uch as ~ ,~ CO ~ rl ~- r,~ l T ~ r,~
fluorida, p~ x~di~ and bicarbqnate a ~ ~, and cations ~uch as silver~ zinc~ mèrcury~ ~rsenic~ copper,. ~ atin~m, antimon~, gold, thallium, nickel, selenium, bi~muth, and ca~mium cation~.
A to~ic ion is one which when introduced into a target cell inhibits and/or pre~ents and/or destroys the growth of the target cell.
Such a toxic ion is one which in the ab~ence o~ an ion channel forminR peptide is unable to cro~s a natural or ~ynthetic lipid membrane; in particular a cell membrane, in sufficient amounts to affect a cell adversely.
Tha pQptide and the to~ic ion, whether administered or prepared in a singla composition or in s~parate compositions, are employed in amounts effective to inhibit and/or prevent and/or destroy tho growth of the target cell. In affcct, the ion potantiates the action of the peptite, i . Q ., the amount of toxic ion i9 effectiv~ to rsduc~ ~he ma~lmum cffectiv~ concentration of thc peptid~ or protein for inhlbitlng ~rowth of a target cell.
Antibiotic~ which may be employ~d in combination with the peptides of the pre~nt invention includ~ b~citr~cins, aminoglyco~ide8, hydrophobic ~ntibiotics, lnclutln~ macrolide antibiotics, penicillins, monobactam~, or deri~atives or analogues thereof.
Other antibiotic~ which may be u~ed (wh~ther or not hydrophobic) in combinstion with the p~ptlde~ of the present in~ention are ant$biotic~ which are 50-S ribosome inhlbitors such as linco~yci~; cllnda~ycin; and chlora~phenicol; etc.; and antibio~ic- whlch have a lsrge lipld li~e lactone ring, such as mystat~n; pi~aricin, etc.
Th~ p~ptid~ and antib$otic may be ad~in~tored by direct administration to a tar~t cell or by ~y~temlc or top~cal administration to a host which lnclud~ the targat cell, in order to pre~ent, de~troy or inhibit th~ ~rowth of a targ~ c~ll.
Tar8et cells whos~ growth may bo pr~onted, inhibited, or de~troy~t by thc admlnistrstion of th~ p~ptld~ ant sntlbiotic :- , , ~ ,Y~ T ~ 2 ~ ~ ~
include Gr~-posItiv~ and Gram- 2 bacteria as w~ll aQ
fungal cells.
The in~entlon will now be furthar described with respect to the following examples; however, tha acope of the present invention i9 not to be llmited th~r~by.

EXAMPLE
The procedure ~or the following antibacterial a3say is based upon the guidelines of the Nstional Committee for Clinical Laboratory Standards, Document M7-T2, Volume 8, No. 8, 1988. ~ l~b A stock solution of 8 p~ptlde of th~ followi ~8 struc~ al formula:
KIAGKIARIAGKIAKIAGKIA-smide, ~ Sl ~
wherein each amino acid re~ldue i8 a D-amino acit re~idue or glycine, was prepsred ~ a concentratiQn of 512 ~g/ml in sterile deionizet tistill2t water ~nd atored at -70C.
The stock peptid~ 301ution ia dllut~d in a~rial dilutions (1:2) down the wells of a microtiter platQ ~o that the final concentrations o~ peptide~ in th~ w~ ar¢ 0.25, 0.50, 1, 2, 4, 8, 16, 32, 64, 128, ~nd 256 I~g/ml. 1.5 ~1 105 CFlJ~/ml of elther S. aureus ATCC 25923, E. coli ATCC 25922, or P. aeru~inosa ATCC
27853 were added to the wells in full strength Mueller Hinton broth (B~L 11443 ~ from a Islid- loE~ cultur~ . Th~ inoculum i8 standarized spectrophotom~trically at 600nm and i8 ~ertified by colony counts. Th~ platQa ar~ incubated or 16-20 hour~ a~ 37C, and thQ ~ini~al inhibitory concentration (MIC~ for each peptide is tat~r~in~d. Minimal inhibitory conc~rltration is d~fin~d as the low~at conc~ntration of peptid~ which p~oduce~ a clea~ well in the mlcrotitar plat~.
Th~ ~lni~l inhi~itory concontrat~on o th~ p~ptid~ was 8 ~/ml a8sin~t S. aureus, 32 ~g/ml a~aln~t P. aeru ino~a, ant 4 l against ~. coli. For compari~on, a atoc~ ~olution o~ the above-mentioned peptid~ wa~ prepar~d as h~rainabov~ d~cribet wherein each amino scid r~idue wa8 not ~ D-amlno acid resldue~

, .

~CeQ ~ 2 ~ 8 This peptid~ wa s~ed~as ~ bove de~cribed f4r M~C
against S. a~eùs,- P. aeruRinosa, and E. coli. The MIC o~ the peptide was 8 ~g/mi a8ainst S. aureus, 16 ~g/ml agai~st P.
aeruRinosa, and 4 ~g/ml against E. coli.
The above result~ indicate that wh2n ~11 residues of biologically sctive peptide are D-amino scid residues or glycine resldues, the peptide retains it~ biological actlvity.
The peptid~9 of the pre~nt invantion, whether sdministered alone or in combination with agents s~ch as to~ic ions, antibiotics, or other biolo~lcally activ~ peptides or proteins as hereinabove d~scrlbed, m~y be employed in a wita variety of pharmaceutlcal compositions in comblnatlon with a non-tox~c pharmaceutica~ carrier or vehiclQ such a~ a filler, non-to~ic buffer, or physiological ~aline ~olution. Such phsrmaceutical composition~ mQy b~ used topic~lly or ay~temic~lly and m~y be in ~ny suitabl~ form ~uch a~ a liquid9 ~olid, ~emi-solid, inJectable 301ution~ tablet, ointment, lotion, pa~te, cap~le or the like.
The peptide and/or a8ent a~ hereinAbove de~cribad may also be u~ed in combination with ad~uvant~, protea8e inhibitor~, or compatible drug~ where such 8 combinatlon i~ ~e~n to be desirable or advanta~eous in controlling inf~ction caused by harmful microorganism~ including protozoa, viru~es ~ and the lik~.
The peptide may be admini~t~rd to a host, in particular an animal, in an eff~ctive antibiotic ant/or anti-tumor and/or an~iviral snd/or a~ti~icrobial and/or anti~permicidal and/or antipara~itic a~ount, or in ~n amount ef~ctive to stimulate wound hQaling ln a host. The pRptid~ may b~ admini~terd ~ither alone or 1~ co~blns~io~ with a to~ic lon, antibiotic, or ion channel for~in~ p~ptide or pro~ein ~a horeinabove described.
When the peptlto i~ atmlni~tered in co~blnation wlth a toxic ion, ths actlvity of tha peptid~ is potentiat~t.
When tha p~ptidQ 18 adminiatQr~d in combination with an agent as h~reinabove de~cribed, 1~ i~ pos~ible to administer the peptide and agent in aepar~te ~orm~. For e~mpl~, the agent msy C Q
be admini~t~rëd 8y8;temically and tho pe~id~:may bs admini~tered topically.
When the peptide i~ adminitared topioally, it may be admini~tered in combination with a wat2r-301uble vehicle, said water-~oluble vehicle bein~ in the form of an ointment, cream, lotion, paste or the like. E~amples of water-~oluble vehicles which may be employed include, but arc not limited to, glycol~, 9UC~ as polyethylen~ glycol, hydroxyc~llulos~, and KY J~lly. The water-~olubl~ vehicle is preferably free of an olly sub~tance.
The peptld~ may slso be employet alone, or in co~bination with other p~ptide~ of the p~e~ent invention, or in combinatlon with peptides containing L-amino acid residues, or in combination with othar active component~ (eg., to~ic ions, ~ntibiotic~) as herainabove described in the form o~ ~n oral composition for oral hygiene. Such a composition mBy be lncorporated into a wide variety of compo~ition~ and matQrials used for oral hy~iene purpose~, which include, but ar~ not llmit~d ~o, toothpastes, mouthwashe~, tooth gels, snd tooth powd~r~. Such compo~ltion may thus be u~et to treat or pr~vent p~r~odontal di~ea~, to prevent or reduce plaque, and/or to prevent or tr~at or r~duc~ dental csries. The peptide may be u~ed to inhibit, preYent, or destroy the growth of 5trePtococcu~ mutans, whlch i9 a~sociated with dental carle~ ant per$odontsl di~as~.
Numerou~ modification~ ant vari~t~on~ of the present invention are po~siblQ ln light of thQ above teaching~ and, therefore, within tho 8COp~ of the a~companying claims, the invention ~ay b~ practiced other than ~ psrticularly described.

Claims (65)

1. A compound comprising an analogue of Magainin I peptide or Magainin II peptide, said Magainin I peptide or Magainin II peptide being in an amide-or carboxy-terminated form, and wherein Magainin I is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residue refer to the position of the residue in the peptide:
G I G K F L H S A G K F G K A F V G E I M K S
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23, and wherein each amino acid residue is a D-amino acid residue or a glycine residue, and wherein Magainin II is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residue refer to the position of the residue in the peptide:
G I G K F L H S A G K F G K A F V G E I M N S
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23, and wherein each amino acid residue is a D-amino acid residue or a glycine residue, and wherein said Magainin I or Magainin II
peptide is substituted in at least one of positions 1-23, wherein the substituents which may be employed in each of positions 1-23 are shown in the following table:

Residue No. Substituent 1 D-Lys,D-Ala

2 D-Lys,D Ala,D-Ile,D-Arg,D-Leu,D-Val,D-His,D-Met

3 D-Lys,D-Ala,D-Trp,D-Arg,D-His

4 D-Lys,D-Ala,D-Arg,D-His D-Phe,D-Ala,D-Lys,D-Trp,D-Leu,D-Ile,D-Val,D-Met 6 D-Leu,D-Lys,D-Ala,D-Ile,D-Val,D-Met 7 D-Lys,D-His,D-Ala,D-Arg,D-Ile,D-Val,D-Met 8 D-Ala,D-Lys,D-Ser,D-Trp,D-Met,D-Ile,D-Arg,D-His, D-Thr,D-Leu,D-Val 9 D-Lys,D-Ala,D-Trp,D-Arg,D-His,D-Leu,D-Ile,D-Val D-Lys D-Ala,D-Trp,D-Arg,D-His,D-Leu,D-Ile,D-Val 11 D-Lys,D-Arg,D-His 12 D-Phe,D-Lys,D-Trp,D-Arg,D-His 13 D-Ala,D-Lys,D-Trp,D-Met,D-Arg,D-His,D-Phe,D-Leu, D-Ile,D-Val 14 D-Ala,D-Lys,D-Arg,D-His D-Lys,D-Trp,D-D-Ala,D-Arg,D-His,D-Phe 16 D-Ala,D-Lys,D-Phe,D-Ile,D-Val,D-Met,D-Leu 17 D-Ala,D-Lys,D-Val,D-Trp,D-Arg,D-His,D-Met,D-Leu 18 D-Ala,D-Lys,D-Trp,D-Arg,D-His,D-Leu,D-Met 19 D-Ala,D-Lys,D-Glu,Gly,D-Arg,D-His,D-Leu D-Ile,D-Phe,D-Asn D-Ile,D-Ala,D-Lys,D-Trp,D-Leu,D-Phe,D-Val,D-Met 21 D-Lys,D-Pro,D-Ala,D-His,D-Leu,D-Arg,D-Ile,D-Phe 22 D-Lys,D-Ala,D-Asn,D-Gln,D-Arg,D-His 23 D-Ser,D-Lys,D-Ala,D-Thr,Gly,D-Leu,D-Ile, D-Gln,D-Asn 2. The compound of Claim 1 wherein said Magainin I or Magainin II peptide is substituted in at least one of positions 1-23, wherein each of the substituents in each of positions 1-23 is shown in the following table:

Residue No. Substituent 1 D-Lys,D-Ala 2 D Lys,D-Ala,D-Ile 3 D-Lys,D-Ala,D-Trp 4 D-Lys,D-Ala D-Phe,D-Ala,D-Lys,D-Trp 6 D-Leu,D-Lys,D-Ala 7 D-Lys,D-His,D-Ala 8 D-Ala,D-Lys,D-Ser,D-Trp 9 D-Lys,D-Ala,D-Trp D-Lys,D-Ala,D-Trp 11 D-Lys 12 D-PhegD-Lys,D-Trp 13 D-Ala,D-Lys,D-Trp 14 D-Ala,D-Lys D-Lys,D-Trp,D-Ala 16 D-Ala,D-Lys,D-Phe 17 D-Ala,D-Lys,D-Val,D-Trp 18 D-Ala,D-Lys,D-Trp 19 D-Ala,D-Lys,D-Glu D-Ile,D-Ala,D-Lys,D-Trp 21 D-Lys,D-Pro,D-Ala 22 D-Lys,D-Ala,D-Asn 23 D-Ser,D-Lys,D-Ala 3. A compound comprising:
an analogue of <againin I peptide or Magainin II peptide 9 said Magainin I peptide or Magainin II peptide being in an amide-or carboxy-terminated form, and wherein Maginin I is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residue refer to the position of the residue in the peptide:

G I G K F L H S A G K F G K A F V G E I M K S
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23, and wherein each amino acid residue is a D-amino acid residue or a glycine residue, and wherein Magainin II is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residue refer to the position of the residue in the peptide:
G I G K F L H S A K K F G K A F V G E I M N S
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23, and wherein each amino acid residue is a D-amino acid residue or a glycine residue, and wherein at least one of amino acid residues 15-23 is omitted, and wherein said Magainin I or Magainin II peptide is substituted in at least one of the remaining positions 1-23, wherein the substituents which may be employed in each of positions 1-23 are shown in the following table:

4. The compound of Claim 3 wherein said Magainin I or Magainin II peptide is substituted in at least one of the remaining positions 1-23, wherein each of the substituents in each of positions 1-23 is shown in the following table:

Residue No. Substituent l D-Lys,D-Ala 2 D-Lys,D-Ala,D-Ile 3 D-Lys,D-Ala,D-Trp 4 D-Lys,D-Ala S D-Phe,D-Ala,D-Lys,D-Trp 6 D-Leu,D-Lys,D-Ala 7 D-Lys,D-His,D-Ala 8 D-Ala,D-Lys,D-Ser,D-Trp 9 D-Lys,D-Ala,D-Trp D-Lys,D-Ala,D-Trp 11 D-Lys 12 D-Phe,D-Lys,D-Trp 13 D-Ala,D-Lys,D-Trp 14 D-Ala,D-Lys D-Lys,D-Trp,D-Als 16 D-Ala,D-Lys,D-Phe 17 D-Ala,D-Lys,D-Val,D-Trp 18 D-Ala,D-Lys,D-Trp 19 D-Ala,D-Lys,D-Glu D-Ile,D-Ala,D-Lys,D-Trp 21 D-Lys,D-Pro,D-Ala 22 D-Lys D-Ala,D-Asn 23 D-Ser,D-Lys,D-Ala

5. A compound, comprising:
a deletion analogue of an amide or carboxy terminated Magainin I wherein Magainin I is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residue refer to the position of the residue in the peptide:

G I G R F L H S A G, K F G K A F V G E I M K S
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23, and wherein each amino acid residue is a D-amino acid residue or a glycine residue, and at least one of amino acids 15 through 23 is omitted.

6. The compound of Claim 5 wherein at least one of amino acids 15, 16, 18, 19, 21, 22 and 23 is omitted.

7. A compound comprising:
a deletion analogue of an amide or carboxy terminated Magainin II wherein Magainin II is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residue refer to the position of the residue in the peptide:
G I G K F L H S A K K F G K A F V G E I M N S
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23, and wherein each amino acid residue is a D-amino acid residue or a glysine residue, and at least one of amino acids 15 through 22 is omitted.

8. The compound of Claim 7 wherein at least one of amino acids 15, 18, 19, 20, 21, and 22 is omitted.

9. A peptide comprising:
said peptide containing at least eight amino acids, wherein each amino acid is a D-amino acid or glycine, said peptide containing at least two groups of amino acids A-B wherein each A-B group is separated from another A-B group by at least one D-amino acid other than a hydrophobic amino acid and at least two groups of D-amino acids C-D wherein each C-D group is separated from another C-D group by at least one D-amino acid or glycine other than a hydrophilic D-amino acid, wherein each of A and B is a hydrophobic D-amino acid or glycine wherein each of A and B may be the game or different amino acids and one of C and D is a basic hydrophilic D-amino acid ant the other of C and D is a basic or neutral hydrophilic D-amino acid.

10. The peptide of Claim 9 wherein the peptide has from 8 to 15 amino acids.

11. The peptide of Claim 10 wherein the peptide includes one of the following sequences:
(i) (Wl)a(A-B-C-D)n(Z1)b (ii) (W2)a(B-C-D-A)n(Z2)b (iii) (W3)8(C-D-A-B)n(Z3)b (IV) (W4)a(D-A-B-C)n(Z4)b wherein W1 is D-, C-D-, B-C-D-W2 is A-, D-A-, C-D-A-W3 is B-, A-B-, D-A-B-W4 is C-, B-C-, A-B-C-Z1 is -A, -A-B, -A-B-C
Z2 is -B, -B-C, -B-C-D
Z3 is -C, -C-D, -C-D-A
Z4 is -D, -D-A, -D-A-B
n is 2 or 3, a is 0 or 1 and b is 0 or 1.

12. The peptide of Claim 9 wherein the peptide contains at least 16 amino acids and there are at least four groups of amino acids A-B ant at least 3 groups of D-amino acids C-D.

13. The peptide of Claim 12 wherein there are at least four groups of D-amino acids C-D.

14. The peptide of Claim 13 wherein the peptide includes at least four groups of the peptide sequence -A-B-C-D, -B-C-D-A-, -C-D-A-B, -D-A-B-C.

15. The peptide of Claim 14 wherein the peptide includes one of the following sequences:
(X1)a(A-B-C-D)n(Y1) (X2)a(B-C-D-A)n(Y2)b (X3)a(C-D-A-B)n(Y3)b (X4)a(D-A-B-C)n(Y4)b wherein X1 is D; C-D- or B-C-D-, Y1 is -A or -A-B or -A-B-C

X2 is A-, D-A- or C-D-A-Y2 is -B, -B-C or B-C-D
X3is B-, A-B-, D-A-B-Y3 is -C, -C-D, -C-D-A
X4is C-, B-C-, A-B-C-Y4 is -D, -D-A, -D-A-B
a is 0 or 1; b is 0 or 1;
and n is at least 4.

16. The peptide of Claim 13 wherein said hydrophobic amino acids are selected from the class keynoting of D-Ala, D-Phe, Gly, D-Ile, D-Leu, D-Met, D-Val, D-Trp, and D-Tyr and D-Cys, said neutral hydrophilic D-amino acids are selected from the class consisting of D-Asn, D-Ser, D-Gln, and D-Thr, and said basic hydrophilic D-amino acids are selected from the class consisting of D-Lys, D-Arg, D-His, and D-ornithine.

17. The polypeptide of Claim 16 wherein at least one of said at least four groups of said four amino acids is of the sequence A-B-C-D, wherein A and B are hydrophobic amino acids, one of C and D is a neutral or basic hydrophilic amino acid, the other of C or D is a basic hydrophilic amino acid.

18. The polypeptide of Claim 13 wherein said polypeptide includes the following sequence:
-(D-Ala-D-Phe-D-Ser-D-Lys)4-

19. The polypeptide of Claim 14 wherein said polypeptide comprises a chain of at least 20 amino acids and no greater than 50 amino acids.

20. The polypeptide of Claim 14 wherein said polypeptide includes five groups of said sequence.

21. The polypeptide of Claim 14 wherein said polypeptide includes six groups of said sequence.

22. The polypeptide of Claim 9 wherein said polypeptide includes repeating groups of said four amino acids.

23. A compound, comprising:
a peptide, said peptide including the following basic structure X:

wherein R1 is a hydrophobic amino acid; R2 is a basic hydrophilic amino acid or a hydrophobic amino acid, R3 is a basic hydrophilic amino acid, R4 is a hydrophobic or neutral hydrophilic amino acid, and R5 is a basic or neutral hydrophilic amino acid, and each amino acid residue of said peptide is a D-amino acid or glycine.

24. The composition of Claim 23 wherein the peptide includes the following structure:
Y - X -wherein X is the basic peptide structure of Claim 2 and Y is R5-; R2-R5-; R1-R2-R5- or R2-R1-R2-R5.

25. The composition of Claim 23 wherein the peptide includes the following basic structure - X - Z-wherein X is the basic peptide structure of Claim 2 and Z

R1; R1-R1; R1-R1-R4; R1-R1-R4-R4; R1-R1-R4-R4-R6;
R1-R1-R4-R4-R6-D-Gln; or R1-R1-R4-R4-R6-D-Gln-D-Gln, wherein R6 is D-proline or a hydrophobic amino acid.

26. The composition of Claim 23 wherein the peptide includes the following basic structure (Y)a - X - (Z)b wherein Y and Z are as previously defined in Claims 24 and 25, a is 0 or 1, and b is 0 or 1.

27. The composition of Claim 23 wherein the peptide has the following structure:
G12S3LG4ALKA5LKIG678LGG9(10)QQ
Where:
1 = F, L
2 = G, A
3 = F, L
4 = K, L
= A, G, T
6 = A, T
7 = H, N
8 = A, M, F, L
9 = A, S, T
= P, L

28. A composition comprising at least one biologically active peptide, said at least one biologically active peptide including a chain of at least 20 amino acids of the structure X10, wherein each of said at least 20 amino acids is a D-amino acid or glycine, and wherein X10 is of the formula:

R11-Rl6-R15-R17-, wherein:
R11 is a hydrophobic amino acid;
R12 is a basic hydrophilic amino acid;
R13 is a hydrophobic or basic hydrophilic amino acid;
R14 is a natural hydrophilic amino acid, or a basic hydrophilic amino acid;
R15 is neutral hydrophilic amino acid, R16 is D-proline or a hydrophobic amino acid; and R17 is neutral hydrophilic, basic hydrophilic or hydrophobic amino acid, said at least one biologically active peptide having no greater than 35 amino acids.

29. The composition of Claim 28 wherein said at least one biologically active peptide has no greater than 30 amino acids.

30. The composition or Claim 28 wherein said at least one biologically active peptide is of the structure:
-R11-R11-X-.

31. The composition of Claim 30 wherein said at least one biologically active peptide is of the structure:
D-Leucine-D-Leucine-X-.

32. The composition of Claim 23 wherein said at least one biologically active peptide is of the structure:
-X-R12-R11-R15-R11-R11-.

33. A biologically active amphiphilic peptide, said peptide including the following basic structure X12:
-[R21-R22-R22-R23-R21-R22-R22]-n, wherein R21 is a basic hydrophilic amino acid, R22 is a hydrophobic amino acid, R23 is a neutral hydrophilic or hydrophobic amino acid, and n is from 2 to 5, and wherein each amino acid residue of said peptide is a D-amino acid residue or a glycine residue.

34. The peptide of Claim 1 wherein the peptide includes the following structure:
Y12-X12, wherein X12 is the basic peptide structure of Claim 1, and Y12 is:
(i) R22;
(ii) R22-R22;
(iii) R21-R22-R22;
(iv) R23-R21-R22-R22;
(v) R22-R23-R21-R22-R22;or (vi) R22-R22-R23-R21-R22-R22.

35. The peptide of Claim 33 wherein the peptide includes the following basic structure:
X12-Z12-, wherein X12 is the basic peptide structure of Claim 1 and Z12 is:
(i) R21;
(ii) R21-R22;
(iii) R21-R22-R22;

(iv) R21-R22-R22-R23;
(v) R21-R22-R22-R23-R21;
(vi) R21-R22-R22-R23-R21-R22.

36. The peptide of Claim 33 wherein the peptide includes the following basic structure:
(Y12)a-X12-(Z12)b, wherein Y12 and Z12 are as previously defined in Claims 34 and 35, a is 0 or l, and b is 0 or 1.

37. The peptide of Claim 33 wher3in n is 3.

38. The peptide of Claim 37 wherein said peptide is of the following structural formula as indicated by the single letter amino acid code.
[KIAGRIA]3.

39. The peptide of Claim 33 wherein n is 2.

40. The peptide of Claim 39 wherein said peptide is of the following structural formula as indicated by the single letter amino acid code:
KIA(KIAGKIA)2KIAG.

41. A biologically active amphiphilic peptide, said peptide including the following basic structure: X14:
R21-R22-R22-R23-R21-R22-R22-R21-R22-R22-R22-R21-R22-R22, wherein R21 is a basic hydrophilic amino acid, R22 is a hytrophobic amino acid, and R23 is a neutral hydrophilic or a hydrophobic amino acid, and wherein each amino acid residue of said peptide is a D-amino acid residue or a glycine residue.

42. The peptide of Claim 41 wherein the peptide includes the following structure:
Y14-X14, wherein X14 is the basic peptide structure of Claim 41, and Y14 is:
(i) R22;
(ii) R22-R22;
(iii) R21-R22-R22;
(iv) R23-R21-R22-R22;
(v) R22-R23-R21-R22-R22;
(vi) R22-R22-R23-R21-R22-R22; or (vii) R21-R22-R22-R23-R21-R22-R22.

43. The peptide of Claim 41 wherein the peptide includes the following structure:
X14-Z14, wherein X14 is the basic peptide structure of Claim 41 and Z14 is:
(i) R21;
(ii) R21-R22;
(iii) R21-R22-R22;
(iv) R21-R22-R22-R23;
(v) R21-R22-R22-R23-R21;
(vi) R21-R22-R22-R23-R21-R22; or (vii) R21-R22-R22-R23-R21-R22-R22.

44. The peptide of Claim 41 wherein the peptide includes the following basic structure:
(Y14)a-(X14)-(Z14)b, wherein Y14 and Z14 are as previously described in Claims 42 and 43, a is 0 or 1 and b is 0 or 1.

45. The peptide of Claim 44 wherein said peptide is of the following structural formula as indicated by the single letter amino acid code:
KLASKAGKIAGKIAKVALKAL.

46. The peptide of Claim 44 wherein said peptide is of the following structural formula as indicated by the single letter amino acid code:
KIAGKIAKIAGOIAKIAGKIA.

47. A biologically active àmphiphilic peptide, said peptide including the following basic structure X16:

R32-R31-R31-R32-R31-R31-R31-R32-, wherein R31 is a hydrophobic amino acid, R32 is a basic hydrophilic amino acid, R33 is a neutral hydrophilic, basic hydrophilic, or hydrophobic amino acid, and R34 is a hydrophobic or basic hydrophilic amino acid, and wherein each amino acid residue of said peptide is a D-amino acid residue or a glycine residue.

48. The peptide of Claim 47 wherein the peptide includes the following structure Y16-X16, wherein X16 is the basic peptide structure of Claim 47, and Y16 is:
(i) R31; or (ii) R34-R31.

49. The peptide of Claim 47 wherein the peptide includes the following structure:
X16 -Z16: wherein X16 is the basic peptide structure of Claim 47 and Z16 is:
(i) R31; or (ii) R31-R31.

50. The peptide of Claim 47 wherein the peptide includes the following basic structure:
(Y16)a - (X16) -(Z16)b, wherein Y16 and Z16 are as previously described in Claims 48 and 49, a is 0 or 1, and b is 0 or 1.

51. A biologically active amphiphilic peptide, said peptide including the following basic structure X18:

R32-R31-R31-R32-R31-R31-R31-R32-(R36)n-R31-, wherein R31 is a hydrophobic amino acid, R32 is a basic hydrophilic amino acid, R33 is a neutral hydrophilic, basic hydrophilic, or hydrophobic amino acid, R34 is a hydrophobic or basic hydrophilic amino acid, R35 is glutamine or asparagine, or a basic hydrophilic, or hydrophobic amino acid, ant R36 is glutamic acid, aspartic acid, a hydrophobic amino acid, or a basic hydrophilic amino acid, ant n is 0 or 1, and wherein each amino acid residue is a D-amino acid residue or glycine.

52. The peptide of Claim 51 wherein said peptide includes the following structure:
Y18 - X18, wherein X18 is the basic peptide structure of Claim 51, and Y18 is:

(i) R31; or (ii) R34-R31.

53. The peptide of Claim 51 wherein said peptide includes the following structure:
X18-Z18, where X18 is the basic peptide structure of Claim 51, and Z18 is:
(i) R31;
(ii) R31-R35; or (iii) R31-R35-D-Proline; or (iv) R31-R35-D-Proline-R32.

54. The peptide of Claim 51 wherein the peptide includes the following basic structure:
(Y18)a - (X18) - (Z18)b, wherein Y18 and Z18 are as previously described in Claims 52 and 53, a is 0 or 1, and b is 0 or 1.

55. A process for inhibiting growth of a target cell or virus, comprising:
administering to a host an effective amount of the compound of Claim 1.

56. A process for inhibiting growth of a target cell or virus, comprising:
administering to a host an effective amount of the compound of Claim 3.

57. A process for inhibiting growth of a target cell or virus, comprising:
administering to a host an effective amount of the compound of Claim 5.

58. A process for inhibiting growth of a target cell or virus, comprising:
administering to a host an effective amount of the compound of Claim 7.

59. A process for inhibiting growth of a target cell or virus, comprising:

administering to a host an effective amount of the peptide of Claim 9.

60. A process for inhibiting growth of a target cell or virus, comprising:
administering to a host an effective amount of the peptide of Claim 23.

61. A process for inhibiting growth of a target cell or virus, comprising:
administering to a host an effective amount of the composition of Claim 28.

62. A process for inhibiting growth of a target cell or virus, comprising:
administering to a host an effective amount of the peptide of Claim 33.

63. A process for inhibiting growth of target cell or virus, comprising:
administering to 8 host an effective amount of the peptide of Claim 41.

64. A process for inhibiting growth of 6 target cell or virus, comprising:
administering to 8 host an effective amount of the peptide of Claim 47.

65. A process for inhibiting growth of a target cell or virus, comprising:
administering to a host an effective amount of the peptide of Claim 51.