CA2045073A1

CA2045073A1 - Composition and treatment with biologically active peptides and antibiotics which inhibit dna gyrase

Info

Publication number: CA2045073A1
Application number: CA002045073A
Authority: CA
Inventors: Barry Berkowitz
Original assignee: Individual
Current assignee: Magainin Pharmaceuticals Inc
Priority date: 1990-06-27
Filing date: 1991-06-20
Publication date: 1991-12-28
Also published as: JPH05507718A; WO1992000090A1; EP0559647A1; EP0559647A4

Abstract

ABSTRACT OF THE DISCLOSURE

A composition comprising at least one biologically active amphiphilic peptide or protein, said peptide or protein being an ion channel-forming peptide or protein, and an antibiotic which inhibits DNA gyrase. The biologically active amphiphilic peptide and the antibiotic which inhibits DNA gyrase may be administered in amounts effective to inhibit growth of a target cell such as a bacterium.

Description

PATAP410 ~ 7 3 COMPt)SITION AND TREATMENT WITH BIOLOGICALLY
ACTIV~ PEPTIDES AND ANTIBIOTICS WHICH XNHIBIT
DNA GYRASE

This invention relates to biologically actlve peptldes and proteins, and more particularly to compositiorls and uses ~nvol~g biologically active peptides or protelns and an antlbiotic which inhibits DNA gyrase, and in particular quinolone antibiotics such &S ciproiloxacin.
In accordance with an aspect of the present in~vention, there is provided a composition which includes lncludes at least one biologically active amphiphilic peptide and/or biologically active protein; and an antibiotic which inhlblts DNA gyrase.
In accordance with another aspect of the present invention, there is provided a process wherein there ls administered to a host at least one blologically active amphiphilic peptide which i~ an ion channel forming peptide andJor biological~y active protein; and an antibiotic which inhibits DNA gyrase.
An ~on channel-îorming peptlde or protein or ionophore is a peptide or protein which increases the permeabillty for ionc across a natural or synthetlo llpld rnembrane. B. Christensen et al. PNAS Vol. 85 Pgs.
5072-76 (July, 19R8) descr{bes methodology which indicates whether or not R peptide or proteln has ion channel-forming properties and is therefore an ionophore. As used herein an ion channel-forming peptide or ion channel forming protein is a peptide or protein which has ion chlmnel-for~ming propertie~ as determlned by the metho~i ot Chrlstensen et ~1.

, , . .: .: , ,. ., , ,~

An amphiphlllc peptlde i5 a peptlde whlch includes both hydrophobic and hydrophillc peptlde reglons.
In accordance with an aspect of khe present invention whereln the biologieally actlve peptide or proteLn, and an antlbiotlc which lnhlbits DNA gyrase are admlnlstered to a host, such blologlcally actlve peptide or proteln and the antiblotlc whlch inhlblts DNA gyrase may be admlnlstered as a single compo.sitlon or in separate compositions, and the single or ~eparate composition may include additional materials, actives and/or lnactlves, in addition to the peptide and/or protein and antibiotlc whlch inhiblts DNA gyrase.
In accordance with another aspect, there is provided a use of a composition to lnhlblt growth of a microbe, sald composition comprising:
at least one blologlcally actlve amphilllc peptide and/or biologically active proteln, said peptlde or proteln being an ion ehannel-forming peptide or prctein; and an antiblotlc whlch lnhlbits gyrase, salcl components belng ln an amount e~ectlve to lnhlbit growth of a mlcrobe.
The present inventlon also provides a ccmpo~ltlon comprlslng, (a) at lea~t one blologically actlve amphiphLllc peptlde and/or biolo~ically aetive proteln, ~ald peptide or protein bein~ an icn channe:L-~orm:ln~ peptide or ~roteln~
(b) an antlhiot-Lc whlch lnhlblt~ DNA ~yra~e~ and (e) an acceptable pharmaceutlcal c~rrler, wherein sald eomponenks (a) and (b) are pre~ent in a combinec~ ~mount e~ective to in~l:lb.it growth of a mierobe.

.
~, , ; - ..

2a 6~975-96 The lon channel-forming peptides employed in the present lnventlon are generally water soluble to a concentratlon of at least 20 mg/ml at neutral pH ln water. In addltlon, such peptldes are non-hernolytlc7 l.e., they will not rupture blood cells at effectlve concentratlons. In additlon, the structure of such peptlde provldes for flexlbillty of the peptlde molecule. When the peptlde is placed ln water, lt does not assume an amphlphillc structure. wnen the peptide encounters an olly surface or membrane, the peptide chain folds upon ltself into a rod-llke structure.
In general, such peptldes have at least 16 amlno aclds, and preferably at least 20 amino aclds. In most cases, such peptides do not have ln excess of 40 amino aclds.
DNA gyrase is an enzyme whlch is involved ln the formation of bonds between individual coillng strands of replicating bacterial DNA. Thus, DNA gyrase is necessary for the normal repllcatlon of bacterlal DNA, and, therefore, antlbiotlcs whlch inhiblt DNA gyrase inhiblt the normal replication of bacterial DNA.

. .
., ` `` ` , . : :,` `.~ , .

2 ~ 3 Ex~nples of antibiotics whlch inl~bllt DNA gyrase include nalidixic acid, oxolinic acid, c~oxacin, and qulnolone antibiotics which ~clude ciprofloxacln, nor~loxacin, ofloxacin, enoxac~l, pefloxacin, lomenoxac~, fleroxacln, tosu~loxacin, temafloxacin, and ru:Eloxacin. The îollowing are structural io~mulae of representative ex~nples of sntibiotics which ~nhibit DNA gyrase.
Nalidlxic acid has the followlng structure:
~C~

Oxolinic acid has the followlng structure:

~CoOft Of the antlbiotlc3 whlch Inhiblt DNA gyrase which are also quinolone antib~otics, the followlng are repre~entative structural formulae.
Ciprofloxacin has the followlng structure:
"" ~C~Ofl' NorlEloxacln has the Eollowlng structuro:

U~

' 2 ~ 3 Antibiotic3 whlch inhibit DNA gyrase are further ds~cribed in Clin~c~l and Infectiou~ Diseaseq, W. B. Saunders Co. (1987~ . In a preferred embo~iment, the antib~otlc which inhibitq DNA gyrase is a quinolone antib~otic, and most preferably, the quinolonP antibiotic is cipronoxac~n .
In employlng both an lon channel-form~ng biolog~cally active amphiphllic pepffde or an ion channel-forming protein, and an antibiotic which inhlbits DNA gyraseJ whether admintstered or prepared in a single composition, or in separate compositions, the peptide or prot~in and the antibiotic which inhibits DNA gyrase are employed In announts effective to inhibit andJor prevent and/or destroy the growth of a target cell. In effect, the quinolone antibiotic potentiates the action of the peptide or prote~n, and the peptide or p~oteln potentiates the action of the antibiotic whlch inhibits DNA gyrase . The term "potentiate, " as employed herein, mean3 that the amount of antiblotic which inhibitc DNA gyrase is effective to reduce the minimum effective concentratlon of the peptide or protein for inhibiting growth of a target cell arld the amount of peptide or protein i9 effective to redl~ce the minimum effective concentration of the antibiotic whlch ~nhibits DNA gyrase for inhlbit~ng growth of a target cell.
In gener~l, the peptide or protein is administered topically at a concentration of from .Q5~ to 109~. When adminl~tered systemically, the peptlde or protein is employed to provide paptid~ or proteln dosag~s of from 1nng to 50ûm~ psr kilogram of host welght.
Th~ antiblotlc which Inhiblt~ DNA ggrase, In gQneral, ls used toplc~lly at ~ concentra.tlon of from O.O5~b to 109~ en u~ed systemically, 2 ~ 7 -3 the antibiotic which inhibits DNA gyrase ls generally employed in an amount of from 1.25 to 45mg per kilogram of host weight per day.
The use of a combination o peptide or protein and antibiotic which inhibits DNA gyrase, in accordance with the present inventlon is effective as an antibiotic, and may be employed to inhibit, prevent or destroy the growth or proliferation of microbes, such as bacterla.
The compositions have a broad range of potent antibiotic activity against a plurality of microorganisms, including (~r~m-po~tive and Gram- negaffve bacteria . Such compo~tlon~ may be employed i~or treating or controll~g microbial infection caused by organism~ which ar~ sensltive to such compositions. The treatment may comprise admln~ster~g to a host organism or tis~ues acceptable to or affiliated wlth a microbial infection an anti-microbial amount of such peptlde or protein and antibiotic which inhiblt~ DNA gyrase.
The compositions may also be used as preservatives or sterilant~ for materials susceptible to microblal contamlnatlon.
The compositions of the present invention may also be used in the treatment of external burns and to treat and/or prevent skin and burn infectlons. In partieular, the compo~itlons may be used to treat skin and burn infectiona caused by organl3ms ~u~h as, but not limlted to, ~a~ ~nd S~aureus.
Such composltlon~ may al~o be u~ed ln the preventlon or treatment of eye lnfectlons. Such Infectlon~ ma~ e causati by bacterla such as, but not llmlted to, P ~u~no3a, S.sueru~, ~nd ~;o~orrhoeae.

tn accordance wlth a prefQrred embodiment, the pepticle used in con~unction with the antibiotic whlch lnhiblts DNA gyrase ~9 a basic , . . . ~ ~ .

2 ~ ~ ~ !7 ~

(pocitively charged) polypeptide hav~ng at least sixteen amino aoid~
wherein the polypeptide include~ st lea~t eight hydrophoblc amino acids and at least elght hydrophilic amino acid3. Still more particulasly, the hydrophobic amino acid~ are in groups OI tWID adJacent amlno acid~, and each group of two hydrophoblc amlno acld~ 1~ spaced from another group of two hydrophoblc amlno acid~ by at least one amino acld other than a hydrophobic am~o acld (preerably at lea~t two amino acids) and generally by no greater than four amino aclds, and the amino acids between pairs of hydrophobic amlno aclds may or mRy not be hydrophllic.
The hydrophillc amino acids are generally also In groups of two adlacent amino acids In which at least one of the tWD alllil10 acids is a basic hydrophilic amino acid, wlth such groups of two hydrophilic amino aclds being spaced from each other by at least one ~nino acld other than a hydrophilic amino acid (preferably at least two am~}o acids) and generally no greater than four amino aclds, and the amlno aclds between pa~r~ of hydrophilic amlno acids may or may not be hydrophoblc.
In accordance wlth a partlcularly preferred embocliment, the polyp~ptide compr~ses a chaln of at lea~t four group~ of amino acids, with each group consi~ffn6 of four amino ElC~/d8. Two vf the four amino acids in each group are hydrophoblc amino acid~, and two of the four amino acid~ In each group are hydrophilic, with at le~st one of tha hydropl~lic P~mlno aeids In each group beinK a bn~ic hydrophil~o amlno acld and the other bein6 a baslc or nautral hydrophllic amino ~cid.
Th~ hydrophobio amlno acld~ m~y bQ select~d from the class con~istlng ut Ala, Cy~, Phe, aly, lle, Leu, Illet, Val, Trp, and Tyr.
Tha neutral hydrophilic amino acids m~y b~ sel~ct~d from the class conslsting of A~n, Gln, Ser, and Thr. The ba~ic hydrophllic amlno aclds .~

2 ~
may be selected from the clas~ con~i~tlng oî Ly~, Arg, Hlls and ornithine (O) .
Each of the groups of four asnlno acicls may be of the sequence ABCD, 8CDA, CI:)AB, or DABC, wherein A and B are each hydrophobic amlno acids and may be the same or d~ferent, one of ~ or D ~s a basic hydrophllic amino acid, and the other OI C or D i~ a basic or neutral hydrophilic arnino acld and may be the same or di~erent. In a preferred embodiment, the polypeptide chaln may comprise 5 or ~ groups of this sequence. In each group, each of A, B, C and D may be the same in some or all o the group~ or may be different in some or all of the groups.
The polypepffde chain preferably ha~ at lea~t 20 am~no acids, and no greater than 50 amino acids. It is to be understood, however, that the polypeptide does not have to oon9i9t entirely o:E the groups described above. The polypeptide may have amlno acids extend~g from either or both ends of the noted groups formlng the polypeptide chain and~o} there may be amino aclds between orle or more of the at lea~t four groups and ~till remaln wlthln the scope of th~ Inventlon.
The group~ OI amino acld~ may be repeatlng groups of amlno aclds, or the umino acid~ in the various group~ mny vary provided that in each group of the at lea~t four groups of amino acids there are two hydrophoblc and two hydrophilic amino acid3 a3 hereinabove noted.
Thu~, In a preferrecl embodiment, the biologically active polypeptide camprlses ~I ch~ln Includlng at lea~t foltr ~roup~ of ~mino ~clds, each contalnin~ ~our amino aclds. Two of the four amlno acids In ench group are hydrophob~c, at lenst one amino ~cld 1~ Iba~ic hg drophillic, and the rem~nlng on~ Ig ba~lc or neutral hydrophlllc, wlth the polypeptide chain 2 ~ ~ .5 0 ~ 3 prefersbly havlng at lea~t 20 ~no acid~ but no greater th~ 50 amino acid~ . .
In one embod~71ent, each of the at leaslt four group~ of amfno acids which are In the peptlde chain i9 of the sequence A- ~3 - C - D, B - C - D - A, C-D-A-B or D-A-B-C wherein A and B are hydrophobic amino acids, one of C or D is a basic hydrophilIc amino acid, and the other of C or D is basic or neutral hydrop}~lic amino acid. The s~e~ulting polypeptide chain, there~ore, may have one of the followlng sequlence~:
(Xl)a(~~ B-C-D)n(Yl)b (X2)a(B-C-D-A)n(y2)b (~3)a(C~D~A~B)n~Y3)b (g4)a(D~A~B~C~n(Y4~b wherein Xl is D; C-l:~- or B-C-D-, Yl is -A or -A-B or -A B-C
X2 is A-, D-A- or C-D-A-Y~ is -B, -E3-C or B-C-D
X3is B-, A-B-, D-A-E~-Y3 lc -C, -C-D, -C-D-A
X4is C-, B-C-, A-B-C-Y'4 is -D, -D-A, -D-A-E~
a i3 o or l, b is o or 1 and n is at lea~t 4.
It is to be understood that the peptide chain may include amino acids between the hereinabove noted ~roups of four amino acld~ provided that the ~paoing~ betwaen such groups allci th~ char~ on th~ amlno aclds does not ohange th~ charao~rlstic~ of tha paptlda chaln whlch provicle amphiphiliqlty and a poslti~re charge and do not adv~er~eiy affact the folclln~ aharac~ristic~s of the chain to that which 19 slgnlflcnntl)r dlfterent . .

. . , ... ~. : ~ . .~

2 0 ~ ~ 7 3 from one in which the hereinabove noted group of four amino aclds are not spaced from each other.
As representative example~ of peptide~ in accordance with the pre~ent invention, there may be mentloned.
Ala- Phe - Ser - Ly~ - Ala - Phe - Ser - Ly~ - Ala - Phe - Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser-Ly~
II Al~-Phe- Ser-Lys-Ala-Phe- Ser-Ly~-Ala-Phls- Ser-Ly~ - Ala - Phe- Ser- Lys - Ala- Phe- Ser- Lys - Ala- Ph~-Ser- Ly~ .
III Ph-3- Ser- Lys- Ala- Phe - Ser-Lys-Ala-Phe- Ser-Lys-Ala-Phe - Ser- Lys- Ala-IV Ser-Ly~-Ala-Phe-Ser-Lys-Ala-Phe- Ser- Lys- Ala- Phe- Ser- Lys - Ala-Phe- Ser- Lys- Ala- Phe-V Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser-Ly~-Ala-Phe-Ser-Lys - Ala- Phe- Ser The peptlde, may have amino acid~ extending from either end of the chaln. For example, the chalns may h~ve a Ser-Lys sequencla before the "Ala" end, and/~r an Ala-Phe sequen~e after the "Ly~" end. Other amlno acid sequences may also be attached to the "Ala" and/or the "lys" end.
Similarly, In any polypeptide chaln havlng at least four groups of amino acid~ of the seguence a~ descrtlbed nbove, the chAln may have, for e~sample, a C-D sequence belore the flrst A-B C-D gr~up. Al~o other flmlno acld ~equence~ may be attached to the "A" and/or the "" encl of one ol the~ polypeptlde chain~. Also th~re may be ~mino aclds ln the .

.: .
.. .

,. -10-2 ~ eJ 3 ;

chain which ~pace one or more groups of the herefnabove not~d four amino acids ~rom each other.
The peptides may be produced by known technlques and obtained in substantially pure form. For example, the peptides may be synthesized on an automatic synthesizer. Journal o~ American Chemical Society, Vol.
85 Pages 2149-54~1963). It i~ also poss~ble to produce such peptides by genetic engineer1ng technique~.
In accordancc with another preferred embodiment, the peptide employed ~n con~unction with an antlbiotic which inhibits DNA 8 yrase may be a magainin peptide.
A magainin peptide is either a magainin such a~ magainin I, II or III
or an analogue or derivaffve thereof. The magainin peptides preferably inciude the following baslc peptide structure X12 R11 Rl1 R12 R13 Rll R14 E~12 Rll R14~R12~Rll~Rll~Rll~R14a~ (R15)n~~14a~R14 ~ ~
wherein Rll is a hydrophoblc amino ac~d, R12 is a basic hydrophillc amino acid; R13 i9 a hydrophobic, neutral hydrophllic, or basic hydrophilic amlno acid; R14 and R14a ars hydrophobic or basic hydroph~lic amino acids; R15 is glutamic acld or a~partic acid, or a hydrophoblo or a basic hydrophillc amino aoid, and n is O or 1. In a preferred embodlment, R13 i8 a hydrophobic or neutral hydrophilic amino acid, R14a is a hydrophobic amino aoid, and R15 19 glu~amic acid or aspartic acld.
1'hu~, ~or exampl~, a mag~inin peptide mny Lnolude the following struotur~:
~2 X1 ~, , ,. . ~
,........ . :
-~ ;

2 ~
where 212 is the hereinabove de~crlbed baslc peptide structure and i~
(i) R12 (~) R14a-R12 (iii) R~ 4a E~12 (iv) R14~Rll~Rl~a R12 here ~11' R12' R14 and R14a ~re a~ previously defined.
A magainin peptide may alsn have the fvllourlng stru~ture:
- X - Z
wherein X12 is as prevlou~ly deftned and Z12 is:
(I) Rl~ where RlB is a basic hydrop}~lic am~no ac~d or asp~ragine or glutamine.
(ii) R16-R17 where R17 is a neutral hydrophilio amino acid, a hydrophobic amlno acld, or a basic hydrophilic amino acid. Prefera}:ly, R17 is a neutral hydrophilic an~no acid.
A magainin peptide may al80 have the followlng structure:
(Y12)a~ 12 12 b e Xl2, Y12 and Z12 are aY previously defined and a i9 0 or 1 and b 13 0 or 1.
Th~ magalnin peptides mEIy also ~nclude the following basic peptide strtlcture X13:

E714 Rll ~14a ~12 Rll Rll 12 13 11 14 12 Rll E~ll~R12~- wherel~ ,R1~,R13, R14, and R are amlno aold~ a~ h~reinabo~re de~crlb~d.

;, , ., : .: `' ' '~

, . . . .

2 ~ 3 The magainin peptide may al~o include the ~ollowlrsg structure X13-Z13; wherein X13 is the here{n~bnve descr~bed bastc peptide structure and Z13 is (Rll)n (Rllln (Rll)n (R14a)n (R1$~n (~ a)n ~R14)n (R16)n 17 n 11' R14~ R14a~ R1s, Rl~ and R17 ar~ as herelnabove descrlbed, and n is O or 1, ~nd each n may b~e the ~ame or dlf~erent.
The magainin peptides generally lnclude at least fourteen an~no ac{ds and may include up to forty amino acids. A magainin peptide preferably has 22 or 23 amlno acldc~ Accordingly, the hereinabove described basic peptide structure~ of a rnaga:lnin pept:lde may lnclude addltional alrino acids at the amino end or at the csrboxyl end, or at both end~.
As representative examples of such magainirl peptldes, there may be mentioned peptides having the following primary ~quence (expressed as a single letter code) a~ well a~ approprlate analoglle~ and derivatives t31ereof:
(a) (NH2) GIC:KFLHSA(::KFGKAFVCEIMKS(OH) or (NH2) (Magain~n I) (b) (NH2) GIaKFL}lSAKX~FGXAEVaEIMNS~OH) or (NH2) (Magain~n II) (c) (NH2) alGKFLHSAKKFGRAFVGEIMN(OH) or (NH2) (Maga~nin III) The followlng are example~ of peptlde derivatlve~ or analog~ of the ba~lc ~tructure:
(d) ~NH2) IGKFl.HSAKKF~K~VGEIl!flNS(OH) or (NH2 (~) (NH2) C~KFLlllSAKKFa~AFV~13I~(OH) or (NH~) (IE) (N}12) KFLHS~KK~aKlU@VaEIMNS(OH) or (NE~:2) , , :- .
, 2~ V73 hqagainin peptide~ ar~ described In Proc. Natl. Acad Sci. Vol. 84 pp.
5449-53 (Aug. 87). The term "magainin peptldes" as used herein refer~
to the basic magalnln structure a~ well ~s derivatives and analogs th~reo~, including but :not llmlted to the representative derivatlve~ or an~lOg~q~
In accordance with a further embodim~nt, the peptide employed in conJunction wtth an antib~otic which inhib~ts DNA gyras~ may be a PGLs peptide or an XPF peptide.
A P(~La peptide is either PGLa or an analogue or derivative thereof.
The P~La peptlde~ preferably include the followlng bas~c peptide structurs X14:
- Rll-R17-R12-Rll-R14-Rl~-Rll-R11 ~14 R12 Rll R11 R12 R1l Rll Rll R12 e Rll, R12, R14, and R17 are as prevtously deflned.
The PGLa peptides generally ~nclude at least seventeen amlno acids and may include as many as forty amino acids. Accordingly, the hereinalbove de~crlbed basic peptide ~tructure for a PaLa peptide may lnclude addltional amtno actds at the amino ~nd or at the carboxyl end or at both the amtno and carboxyl end.
'rhu9, tor example, a PC3La pepttde may have the following strllcture:

where X~ q as prevtously cle!~ d and ~14 lg (I) ~1;
~U) R14~

20~t.~.073 where Rll and R14 ara as pre~ou~ly defined.

For example, a PGLa peptide may also have the ~ollowlng structur2:

where X14 i~ a~ previously defined; and Z14 i~:

(~) Rll; or (i~) Rll-Rll wher~ R~ a~ previously de~ined.
A P(~LQ peptide may also have the fo310wi:ng structure:

(Yl4)a 14 ( 14)b where X14; Y14 and ~14 are as previously defined, a i~ û or 1 and b is O or 1 .
An XPF peptide is either XPF or an snalogue or derlvative thereof.
The XPF peptides preferably include the following ba~ic peptide structure xl~
- - Rl 1 ~ Rl - R - ~ - R - R - R
Rll-Rl4-Rl2-Rll-Rll R12 Rll Rll Rll~R12~(Rls)n~RIl~-, whereln Rll' R12' E~14~ Rls and E~17 are a~ prevlou91y defilled and R18 is glutamine or aspuragine or a basic hydrophillc, or hydrophobic amino acid and, n i~ O or 1.
The XP~ peptldes generally include at least ninateen amino acids and may lnclude up to l~orty am~no acld~. Accord:lngly, the herelnabove do~cribed ba~io pepttdo ~tructUro of XPlF may Include addltlonal amino ~cld~ ~t the amlno end, ~r at the carboxyl end or at both the amino and carbo~syl end~.

., , .:. : . :
. ~ , .
2~'3~3 Thll~, ror exampl~, an XPF peptlde may ~nclude the followlng structure:
-Yl~-X16-where X16 1~ ac previously defined and Y

(1) R11 or (ii) R14-Rll where Rll and R14 are ag pre~ou~ly de~ d.

An XPF peptide may include the following structure:
- X ~ Z 1 where Xl~; is as previously definsd and Z16 i~
(~) R11; or (~i, R11-R18; ~r (~) Rl1-R18-Proline; or (iv) Rll-R18-Pro~g R12 An XPF pept~de may also ha~e the following structure:
(Yl~)a X16(Zl~)b w~ere X16, Y1~ and Zl~ are a~ prevlously defined: a i~ 0 or 1 and b is 0 or 1.
Pref~rred are XPF or PGLa peptld~s, which are characterllzed by the rOIlOwlng primary amino scid ~equenc~ ngle letter amino acid code):
P~L~: GMASKAGAIAGE~IAK'VALKAL (NH2) XPF: aWASXIGQTLGRIAXVaLKELIQPR
A review of XPF and PC~La can b~ folmd In Hoffm~n et al, EMB0 J.
2:711-q14f 1983; Anclreu ~t ~l, ~ hQm. 149:531-535, IgB5; (:~ib3cn ~t al J. Blol. ~ hem. 2~1:53~1-5349, 198~; ~nd Gio~annlnl ~t a1, Blochem J.
~43:113 120, 1~87.

, r 2 ~ J' 3 In accordance wlth yet another embod~nent, the peptide employed ~n con)unction with an antibiotlc whlch inhiblts DNA gyrase may be a CPF
peptide or appropriate analogue or derv~ative thereof.
CPF peptlde~ as well 89 analogues and derivatives thereof are herein sometimes referred to collectively as CPF pept~des.
The CPF pept~de is preferably one which include~ the follow~ng peptlde structure X30:

-R -R -R23-~2l-R2l-R24-R2s-R2l-wherein R21 i5 a hydrophobic am~no acid;
R22 is a hydrophobic amino acld or a basic hydrophilic amino acid;
R23 ~s a basic hydrophiiic amino acid; and R24 is a hydrophoblc or neutral hydrophillc amino acld; and R25 is a basic or neutral hydrophilic amlno acid.
The hereinabove basic structure ~9 hereinafter symbolically indicatedas X30.
The hydrophobic amino acids are Ala, Cys, Phe, Gly, IIQ~ Leu, Met, Val, Trp, and Tyr.
The neutral hydrophllic amino acld~ are Asn, Gln, Ser, and Thr.
The basic hydrophilic amino acids are Lys, Arg, His, ancl ornlthine.
Th~ CPF peptlde may include only the hereinabove noted an~no acids or may ~nclude addltlonal amlno acids at the amino end or carboxyl end or bath the amlno and carboxyl end. In ~en~rul, the peptid~ does not lnclude mora th~ 40 flmlnO acld~.

- ~ :

2 ~

~ e CPF peptldes including the above b~slc peptide structure may have from 1 to 4 addit~onal amino aclds at the amlno end. Accordingly, such preferred peptldes may be represented by the structural formula:

wherein ~30 is the hereinabove de~crlbed baslc peptide structure and Y30 i9 (i) R25 . or (il) R22-R25; or (i~i) R21 R22 R2s;
(iv) R22 R21 R22 R25; pre~er y Glye~ne -R21~R22 R25 R21, R22, and R25 are a~ preYiously defined, The carboxyl end of the ba3io peptide structure may also have additional amino acids which n~ay r~nge from 1 to 13 addltional amino acids .
In a pre~erred embodim~nt, the basic structure may have rom 1 to 7 additlonal amlno ~cids at the carboxyl end, which may be represented as follow9:
~C ~O Z30 wherein X30 i9 the hereinabove dQfined ba~lc peptide structllre and Z30 is (~) R21, (~1) R~l-R21-;
21 ~2~;
(1~) R2~-~2~-R24 R24;
(v) R~l R21 ~24 E~24 ~
2~ R2~-~24-R~ R2~ n; or ,:

-2 ~ 7 ~

(~rli) R2l-R2~-R2~-R2~-R2~ G
where~ R21 and R24 are a~ pre~lou~ly defined, and R2~ i~ prollne or a hydrophobic amino acid.
Preferred peptlde~ may be represented by the îollowlng structural formula:
(Y30)a 3~ 3~ b where~n ~30, Y30 and Z30 are a~ previously defined and a is 0 or 1 and b is 0 or l.
Representative examples of CPF peptides wl~ch are u~;eful in the present i3lvention some of whiclh have been de~cr~be~l in the literature and comp~se the following sequences (single letter am~o ~cld code):
( 1 ) GFG SFLGLALKAA1~CIGANALGGAPQQ
(2) GLASFI,GKALKAGLKIGAHLLG~APQQ

(3) GLASLLGKALKA~LKIGTHFLGGAPQQ

(4) GLASLLGKALKATLKIGTHFLGGAPQQ
~S) GFASFLGKALKAALKIG~NMLGGTPQQ
(6) GFGSFLGKALKAALXIGANALGGAPQQ
(7) GFGSFLGKALKAALKICANALG~SPQQ
(8) C;FASF~GKALKAALKIGANLL(3GTPQQ
A review o~ the CPF peptide~ can be found ln Richter, K., Egger, R., and Kr~ 198a) J. EJiol. Chem. 2B~, 3~7B-3û80; Wak~bayashi, T.
K~to, H., and Tachlba~a, ~ 85) Nuclelc Aclds Rasearch 13 , l8l7-1828; alb~on, B . W., Poulter, L., Wllllams, D . ~ ., and Mngglo, J . E .
(198~) J. Blol. Chem. 2~1, 5341-S349.
CPF peptides whlch may be employed In the pr~la3ent lnventlon ar~
reprcsented by the tollowlng ~slngle latter amlno ac1d code):

. . . .
- . , ;.;

Gl2S3LG4ALKA5LKIG678LGG9(lO)QQ
Where: .
F, L
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
lO = P, L
The numbered amino acid~ may be employed as described in any combination to provide either a bas~c CPF peptide structure or an analogue or derivativs. The term CPF pe~tlde includs~ the basic peptide structure a~ well as analog~ or derivatlve~ thereof.
In accordance with st{ll another embodiment, the biologlcally active peptide may 1nclude the tollowlnt baslo strucutre X40:

[~4l Et'42 R42 R43~R4l~R42~~42]~- wherein R41 l5 a bas1c hydrophilic amino acid, R4 ~ ~g a hydrophobic arnino acld, R43 is a neutral hydrophilic or hydrophobic amino acid, and n i~ from 2 to 5.
In one embod~nent, such peptlde may include the follow~ng structure:
Y4~ ,o, wh~raln X4~ 13 a~ r~ln~bove de~oribad, and Y4~ ls:

( i~) R,l~ç - R"~2;
( lii ) ~4 l ~ ~42 ~ R42;

"

2 ~

( 1~) E~43 - R41 ~ ~42 R42;
(v) R42-R43-R41-R42 R42' (~) R -~42-R43-R41-R~2-R42~ whereln R41, R4a~ 43 hereinabove descIlbed In accordance with another embodiment, such peptide may lnclude the followLng structure:

X40-Z40, wherein X4~) 19 a~ herelnabove descrlbed, and Z40 is:
(i) R41;
(li) R41-Rd,2;
(iil) R~ 42~R42;
( iv) R41 ~ R42 ~ R42 43;
( ~) R41 ~ R42 ~ R42 ~ R43 ~41;
(vl~ R41-R~2 -E~42 -R43 ~41 R42 -In accordance another embodiment, such peptlde may include the followlng structure:

(Y40)a X40 (~40)b, wherein Y and Z are a~ previously defined, a is 0 or 1, and b i~ 0 or 1.
In one embodiment, n is 3, and most preferably the peptlde is of the following structur~ AS indtcated by the ~Ingle letter amlno acid code:
~ KIA(:~KIAl 3, Ila another embodllnent, n l~ 2, ~nd the peptide preferably 13 of the following structur~ as ~ndicated by the ~qtngle letter ~r.lno acid code:

KIA(KIAC~KIA)2KIA~.
In accordanca wlth yet anot~er embod~ment; the blolo~lcally actlve amphlphlllo peptIde atn~ be n bloto~loally aotive amphlphlIIc peptkle InoludinK the foUowin,~ b~lo ~tructur~ ~50:

' . ' 2 ~ 7 3 R41 ~42 R42-R~3-R41-R42-R42-~41-R42-R42-R42-R41-R~-R42, wh~rein R41, R~2 and R43 sre ax here~nabove described.
In accordance u~th one embod~ment, such peptide may include the fo~ow~ng structure:
Y50-X50, wherein X50 is a~ hereinabove ~escribed, and Y50 ~s:
(I) R42;
~) R42 R42;
~) R41-R42 R42;
(lv) R43-R41-R42 R42;
(v) R42-R43-R4l R42 R42;
(vl) ~42-~42-R43-R41-R42 R42- ~r (V~) R4l~R42~R42~R43~R4l~R42-R42,Wher~n R41, R42 and 43 as here~nabove desc~bed.
In accordance w~th another embod~nent, such peptlde may include the fo~ow~ng structure:
X5~-Z50, whera~n X50 is as here~n~bove descr~bed and Z50 is:
(i) R41;
~) R41-R42;
(U1) R41-R42- R4a;
(l~) R41-R42-R42 R43;
(~) R~1~R42~R42~R43 R41;
(vi) R41-~42-R4~-R43-R41-R42; or (~U) ~41 R42 ~42-R43-R41-R42-R~Z wh~r~ln R4~, R4~ and R~3 are h~re~nabo~ desartb~d.
In accordanc~ w~th yet another embod~etlt the peptide may ~clude tha foUow~n~ structure:

- -a2-2 ~

(Y50)a-~50-~z5o)b~ wherein X and Y are a~ prevlously defined, a is O or 1, and b i~ 0 or 1. In one embodi7llent , the peptidQ ~9 of the followlng structural formulal as Lndicated by the single letter amino acid code:
KLASKAGKIAGKIAKVALKAL .
In another embodiment, the peptlde i9 of the follo~ng structural formula as indicated by the single letter alslino aclcl code:
KIAG~AKIAGOIAKIAGKIA.
In still another embod~nent, the peptide employed in conjunction with an antlbiotic which inhibit~ I)NA gyra3e is a cecropin. The cecropins and analo~ and derivatives thereof are de~cribed 1n Ann. Rev. Microbiol 1987 Yol. 41 pages 103-2~, in particular p. 108 and Chri~tensen at al PNAS
Vol. 85 p. 5072-76, which are hereby incoxporated by reference.
The term cecropins includes the ba~ic structure as well as analogues and derivative~.
In yet another embodiment, the peptide emlployed in con~unctlon with an antibiotlc whlch inhiblts DNA gyr~se ls a sarcotoxirl. Th~s sarcotoxins and analog~ and deri~ratives thereof are de~crlbed in Molecular Entomo10gy page~ 3~9-78 in p~rffoulnr p. 375 Alnn E~. Li3s Inc. (1987), wl~ch 1 hereby l~ncorporated by reference.
The term s~rcoto~sln include~ the b~slc material~ as well a~ analogues and derivatlves.
It ic also contemplated that wlthln the scope of the present invqnt10n, that each of th~ amino acld residue~ of the bioloe~ically actlve amphlph~lic paptld~ ~truoture~ herelnnbave descrlbad 1~ n D-amino ~cld rq~lcllle or ~ ~lyclne r~1due.

:

'q ~

In another embodiment, an ion channel-forming protein may be used in con~unction wlth an antibiotic which inhibits DNA gyrase. Ion channel-forming proteins which may be employed include defensins, also known as human neutrophil antimicrobial peptides (HNP), ma~or basic protein (MBP) of eosinophils, bactericidal permeability-increasing protein (E~PI), and a pore-formi~g cytotoxin called variously perforin, cytolysin, or pore-forming protein . Defensin~ are described in Selsted , et al ., J .
Clin. Invest. 7 Vol. 76, pgs. 143~-1439 (lg85) . MBP proteins are described in Wasmoen, et al., J. Biol. Chem., Vol. 263, pgs 12559-12563.
(1988). BPI proteins are described in Ooi, et al, J. Biol. Chem., Vol.
262, p~s. 14891-148~4 (1987). Perforln i~ described in Henkart, et al., J. Exp. Med., 160: 75 (1984), and in Podack, et al., J. Exp. Med., 160: 695 ( 1984) . The above articles are hereby incoroporsted by reference.
The term ion channel-forn~ng proteins includes the basic structures of the ion-forlrdng proteins as well as analogue~ and deri~atlYes.
The present invention wlll be further described with respect to the following example; however, the scope of the invention is not to be limited thereby.
_xample 1 Approximately 1 - 5 x 105 colony forn~ng units (CFU's) of P aeru~nosa strain 27853 or of P.aeru~inos~ strain 107 (whlch is gentmnicin - resistant) dispersed in lO0 ul of trypticase soy broth (TSB) were ndded to a~ch ~,e a series of te~t well~. Either P~ptide 1, Peptide 2, or Peptlda 3 wns ndded to each test well ln ~cren~lng amounts from 0.~ to 25~ n ab~anca of or in th~ presenoe of 2~ of the minimal Inhlbitory concentration (MIC) of ciprofloxacln. For purposQs of thls , 2 ~ 3 example, Peptide 1 is amide- terminated Maga~nln II, Peptide 2 is of the follow~ng structural formula:
~ KIAGKIA 3 3; and Peptide 3 i9 of the followlng structural formula:
KLASKAGKIAGXIAKVALKAL .
The MIC of ciprofloxac~n alone against P.ae~g~nosa straln 27853 was l~g/ml, and again3t P.aeru~nosa strain 10'7 was 2 ~g/ml. The MIC
value~ for Peptide3 1, 2 and 3, elther alone or in combination with 2~ of the MIC of ciprofloxac~n are given in Table I below.
Table I
MIC (~Jg/ml~
P.aeru~o~a strain 1. Peptide 1 alone >32 >32 2. Pepl:ide 1 plu~ 32 32 20~ MIC of cipronoxacin 3. Peptide 2 alone 32 32 4. Peptide 2 plus lB 1 20% MIC o~
ciprofloxacin 5. Peptlde 3 alone 1~ 32 1~. PeptldQ 3 plu~ 8 8 20~ ~1IC of ctpronoxacin Ex~nple 2 For Exampla~ 2 throu~h 4, microorgani~m~ employ~d irl the assays were ~roWn accordlng to th~ procedurc de~cribed In Stutman, et al., Ani~n1crobial A~ent~ ln Chemoth~, ~rol. 3~1, July 1~
In ~ mple~ 2 thrau~h 4, ~ or~anl~m~ were suboulttlred on a~ar plates, and then grvwn ln tryptica~ ~oy broth (r9B), or Mueller-Hlnton , : :, . ~ ~ ... ..
-. . ~ , .
.

2 ~ 3 broth. The final assays were then ecndllcted in microtiter plates containing Mueller-Hinton brot~105 organisms were added to each well.
The minimal Inhlb~tory concentratlons (MIC's~ of ciprofluxac;n alone, of Peptide 1, a~ hereinabove described ~n Example 1, alone, of ciprofloxacin when Peptide 1 was added ( clprofloxacln + Peptide 1 ), and of Peptide 1 when ciprofloxacin was added, ~ Peptide 1 + cipro~loxacin ) were tested against various isolates of strain MR- PSA si~ Pseudomonas aeru~inosa. The second compound (or "~" compound) dose to establish synergy was the lowest dose of the synergizillg drug which alone lacked antibacter~al effect and was at leact a 5~ lower dose than the MIC for the synergizing agent alone. The results are given below in Table 2.
Table 2 hqIC ( I~/mll Ciprofloxacin Ciprofloxacin Peptlde 1 Peptide 1 Isolate one +Peptide 1_ Alorle*ClProfloxacln ~O . 5 <O. ~ 32 8 2 <O. 5 N/A >256 l~/A
3 0 . 25 <O . 25 32 8 1 . O <O . S 255 6~
The minimal inhibitory concentrations, (MIC's), according to the procedure of Example 2, were te~ted agaInst various isolates o~ strain MR-PSA, of Pseudomonas_ aeru~{nosa, e~cept that Peptide 4 replaces Peptide 1. Peptide 4 has the foUowlng structural formula:
GIGKFLKSAKK~GKAFV~KIMNS. The results are glven below in Table 3.
Tab~ 3 Clprofloxac~n ~æoflnxaolnPaptide 4 P~
_aolate~ _Alone ~_ e 4 _~lone ~r~flox~cln , ~:
.. , : ; ~
.~ ' -2~-2 ~ 3 2 C0.5 32 8 2 0.25 0.25 4 4 3 0.25 0.25 8 8 . ` 4 0.063 0.25 8 0.25 0.063 8 4 6 4 ~0.5 32 8 7 0 . 125 0 . 25 16 16 0.5 0.5 16 16 9 2 <0. 5 32 16 0.25 0.25 8 8 11 1 <0 . 125 64 0. S
12 2 <0. 5 32 8 E$amPIe 4 The minimal lnhibitory concentrations (MIC's) of Peptide 4 ancl ciprofloxacin, etther alone or in combination with each other, aocording to the procedure described in Ex~mple 3, were tested agalnst various isolates of St~Phylococcus aureus. The results are given in Table 4 below.
T~ble 4 ClProfloxacin Gi~roiloxacin Peptide 4 Peptide 4 Isolate~ Alone l PePtide 4 Alone~ CiProf10xacin 0. 125 32 8 2 0 . ~5 0. 25 lG 1 3 0. 5 0. 125 32 2 4 >0 . 25 0 . 063 64 32 The pepttde or protein ~nd antibiotic wh~ch inhibit~ DNA gyrase, as hereinabove described, may be employed for treating a wlde varlety of ho~ts. In accordance wlth a preferred embodiment, a ho~t i~ an animal, and suoh aninal may be a h~unan or non-hwnan animal. The pepffde or prot~ln and the antlbiotlc which inhlblt~ DNA gyrase may be employed tog~th~r in a ai~ compo~ltlon, or ln s~p~rate oompo~ttions. ~oreover, the antlblottc whioh Inhlbtt~ DN~ gyra~e and the peptlde or proteln m~ly be dellveroA or ~dminlatered in dlflrerent form~, for example, the . .
. ~
,~

~ ~ ~ r ~ r antibiotic which inh~bits DNA gyra~e may be admlnistered systemically, whlle the peptide or protein may be administered topically.
The peptide or protein and/or antibiotic which inhibits DNA gyrase may be employed in a wide variety o~ pharmaceutical compositions in combination with a non- toxic pharmaceutical carrier or vehicle such as a filler, non-toxic buffer, or physiologicall saline soluffon. Such pharmaceutical compositions may be used topically or systemically and may be in any suitable form such AS a liquid, solid, semi-~olid, in~ectable solution, tablet, olntment, lotion, paste, capsule, or the like. The peptide or protein and/or antibiotic which inhiblts DNA gyrase may also be used in combination with ad~uvants, prot~ase inhibitors, or compatible drugs where such a combination i~ seen to be de6irable or ad~antageous in controlling inîection cau3ed by harn~ul microorganism~, In particular bacteria .
The peptide(s) or protein(s) of the present invention may be admini~tered to a ho~t; in particular an animal, in an effective anti-microbial, in particular in an ant~-bacterial amount, in con~unction with an antlbiotlc which inhibit~ DNA gyrase, for potentiating the acti~ity of the pept~de or protein.
A~ repre~entati~re examples of administering the peptide or protein and antibiotIo which inhibits DNA gyrase for topical or local administratlon, the peptide could be administered in an amowmt of up to nbout 1~ wei6ht to weight And the Antibiotic which inhlbit~ DNA gyra~e clellvered In an ~mount of about 50 mM (~bout 0.1~). Alternatively, the antlbiotic whioh ln}~blts l:)NA ~rrase could be admlni~tered toplcally in conJImction wlth s~ystemic aclmlni~tratiorl Or the peptide and/or protein.
For o~nple, the peptide or proteln may be admini~tered I~l or IP to ' 2 ~ 3 achieve a serum dose of 100 rnicrograms per milliliter tlû miLtlgrams per kilogram) in conjunction with a topical dose of antibiotic which inhibits DNA gyrase of from about 4 IJg/ml to about 100 ~"g/ml.
Nwnerous modificatlon3 and variations of the present invention are possible in lIght of the above teachings and, therefore, within the scope of the appended claims, the invention may be practiced other than as particularly described.

.
, ,: . ~ :'

Claims

1. A use of a composition to inhibit growth of a microbe, said composition comprising:
at least one biologically active amphiphilic peptide and/or biologically active protein, said peptide or protein being an ion channel-forming peptide or protein; and an antibiotic which inhibits DNA gyrase, said components being in an amount effective to inhibit growth of a microbe.

2. The use of Claim 1 wherein the peptide is a magainin peptide.

3. The use of Claim 1 wherein the peptide includes the following basic structure X40:

[R41-R42-R42-R43-R41-R42-R42]n, wherein R41 is a basic hydrophilic amino acid, R42 is a hydrophobic amino acid, R43 is a neutral hydrophilic or hydrophobic amino acid, and n is from 2 to 5.

4. The use of Claim 1 wherein the peptide includes the following basic structure X50:
R41-R42-R42-R43-R41-R42-R42-R4l-R42-R42-R42-R41-R4442-R42, wherein R41 is a basic hydrophilic amino acid, R42 is a hydrophobic amino acid, and R43 is a neutral hydrophilic or hydrophobic amino acid.

5. The use of Claim l wherein said antibiotic which inhibits DNA gyrase is a quinolone antibiotic.

6. The use of Claim 5 wherein said quinolone antibiotic is ciproflocacin.

7. The use of Claim 1 wherein said antibiotic which inhibits DNA gyrase is nalidixic acid.

8. The use of Claim 1 wherein said antibiotic which inhibits DNA gyrase is oxolinic acid.

9. The use of Claim 1 wherein said antibiotic which inhibits DNA gyrase is cinoxacin.

10. A composition comprising:
(a) at least one biologically active amphiphilic peptide and/or biologically active protein, said peptide or protein being an ion channel-forming peptide or protein;
(b) an antibiotic which inhibits DNA gyrase; and (c) an acceptable pharmaceutical carrier, wherein said components (a) and (b) are present in a combined amount effective to inhibit growth of a microbe.

11. The composition of Claim 10 wherein the peptide is a magainin peptide.

12. The composition of Claim 10 wherein the peptide includes the following basic structure X40:

[R41 R42 R42 R43 R41 R42 R42]n wherein R41 is a basic hydrophilic amino acid, R42 is a hydrophobic amino acid, R43 is a neutral hydrophilic or hydrophobic amino acid, and n is from 2 to 5.

13. The composition of Claim 10 wherein the peptide includes the following basic structure X50:
R41-R42-R42-R43-R41-R42-R42-R41-R42-R42-R42-R41-R442-R42, wherein R41 is a basic hydrophilic amino acid, R42 is a hydrophobic amino acid and R43 is a neutral hydrophilic or hydrophobic amino acid.

14. The composition of Claim 10 wherein said antibiotic which inhibits DNA gyrase is a quinolone antibiotic.

15. The composition of Claim 14 wherein said quinolone antibiotic is ciprofloxacin.

16. The composition of Claim 10 wherein said antibiotic which inhibits DNA gyrase is nalidixic acid.

17. The composition of Claim 10 wherein said antibiotic which inhibits DNA gyrate is oxolinic acid.

18. The composition of Claim 10 wherein said antibiotic which inhibits DNA gyrase is cinoxacin.

19. The composition of Claim 12 wherein said peptide is of the following structure:
[KIAGKIA]3.

20. The composition of Claim 13 wherein said peptide is of the following structure:
KLASKAGKIAGKIAKVALKAL.

21. The composition of Claim 11 wherein said magainin peptide is Magainin II.

22. The composition of claim 10 wherein each of said components (a) and (b) are present in an amount ineffective to inhibit growth of a target cell in a host if administered alone to a host.

23. The use of Claim 1 wherein the peptide is a PGLa peptide.

24. The use of Claim 1 wherein the peptide is a XPF peptide.

25. The use of Claim 1 wherein the peptide is a CPF peptide.

26. The use of Claim 1 wherein the peptide is a cecropin.

27. The use of Claim 1 wherein the peptide is a sarcotoxin.

28. The use of Claim 2 wherein said magainin peptide includes the following basic peptide structure:

-R11-R11-R12-R13-R11-R14-R12-R11-R14-R12-R11-R11-RR11-R14a-(R15)n-R14aR14-, wherein R11 is a hydrophobic amino acid, R12 is a basic hydrophilic amino acid, R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid, R14 and R14a are hydrophobic or basic hydrophilic amino acids, R15 is glutamic acid or aspartic acid, a hydrophobic amino acid, or a basic hydrophilic amino acid, and n is 0 or 1.

29. The use of claim 2 wherein said magainin peptide includes the following basic peptide structure:
-R14-R11-R14a-R12-R11-R11-R12-R13-R11-R14-R12-R11-R11-R12-wherein R11 is a hydrophobic amino acid, R12 is a basic hydrophilic amino acid, R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid, R14 and R14a are hydrophobic or basic hydrophilic amino acids.

30. The composition of Claim 10 wherein the peptide is a PGLa peptide.

31. The composition of Claim 10 wherein the peptide is a XPF
peptide.

32. The composition of Claim 10 wherein the peptide is a CPF
peptide.

33. The composition of Claim 10 wherein the peptide is a cecropin.

34 68975-9 34. The composition of Claim 10 wherein the peptide is a sarcotoxin.

35. The composition of Claim 11 wherein said magainin peptide includes the following basic peptide structure:
-R11-R11-R12-R13-R11-R14-R12-R11-R14-R12-R11-R11-RR11-R14a-(R15)n-R14a-R14-, wherein R11 is a hydrophobic amino acid, R12 is a basic hydrophilic amino acid, R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid, R14 and R14a are hydrophobic or basic hydrophilic amino acids, R15 is glutamic acid or aspartic acid, a hydrophobic amino acid, or a basic hydrophilic amino acid, and n is 0 or 1.

36. The composition of Claim 11 wherein said magainin peptide includes the following basic peptide structure:
-R14-R11-R14a-R12-R11-R11-R12-R13-R11-R14-R12-R11-R11-R12-, wherein R11 is a hydrophobic amino acid, R12 is a basic hydrophilic amino acid, R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid, R14 and R14a are hydrophobic or basic hydrophilic amino acids.

37. A use of a composition to inhibit growth of a microbe, said composition comprising at least one biologically active amphiphilic peptide selected from the group consisting of, (a) a magainin peptide;
(b) a PGLa peptide;
(c) an XPF peptide;
(d) A CPF peptide, (e) a cecropin (f) a sarcotoxin (g) a basic polypeptide having at least sixteen amino acids, wherein said basic polypeptide includes at least eight hydrophobic amino acids and at least eight hydrophilic amino acids;

(h) a peptide including the following basic structure X40:
(R41-R42-R42-R43-R41-R42-R42)n, wherein R41 is a basic hydrophilic amino acid, R42 is a hydrophobic amino acid, R43 is a neutral hydrophilic amino acid or hydrophobic amino acid, and n is from 2 to 5; and (i) a peptide including the following basic structure X50:
R41-R42-R42-R43-R41-R42-R42-R41-R42-R42-R41-R42, wherein R41 is a basic hydrophilic amino acid, R42 is a hydrophobic amino acid, R43 is a neutral hydrophilic amino acid or a hydrophobic amino acid, and an antibiotic which inhibits DNA
gyrase, said biologically active amphiphilic peptide and said antibiotic which inhibits DNA gyrase being in a combined amount effective to inhibit growth of a microbe in a host.

38. The use of claim 37 wherein the peptide is a magainin peptide.

39. The use of claim 38 wherein said magainin peptide includes the following basic peptide structure, R14a-(R15)n-R14a-R14-, wherein R11 is a hydrophobic amino acid, R12 is a basic hydrophilic amino acid, R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid, R14 and R14a are hydrophobic or basic hydrophilic amino acids, R15 is glutamic acid or aspartic acid, a hydrophobic amino acid, or a basic hydrophilic amino acid, and n is 0 or 1.

40. The use of claim 38 wherein said magainin peptide includes the following basic peptide structure:

-R14-R11-R14a-R12-R11-R11-R12-R13-R11-R14-R12-R11-R12-, R11 is a hydrophobic amino acid, R12 is a basic hydrophilic amino acid, R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid, and R14 and R14a are hydrophobic or basic hydrophilic amino acids.

41. A composition comprising:
(a) at least one biologically active amphiphilic peptide selected from the group consisting of:
(i) a magnesium peptide;
(ii) a PGLa peptide;
(iii) an XPF peptide;
(iv) a CPF peptide;
(v) a cecropin, (vi) a sarcotoxin;
(vii) a basic polypeptide having at least sixteen amino acids, wherein said basic polypeptide includes at least eight hydro-phobic amino acids and at least eight hydrophilic amino acids;
(viii) a peptide including the following basic structure X40, (R41 R42 R42 R43 R41 R42 R42)n' wherein R41 is a basic hydrophilic amino acid, R42 is a hydro-phobic amino acid, R43 is a neutral hydrophilic amino acid or a hydrophobic amino acid, and n is from 2 to 5; and (ix) a peptide which includes the following basic structure X50:
R41-R42-R42-R43-R41-R42-R42-R41-R42-R42-R42-R41-R442-R42, wherein R41 is a basic hydrophilic amino acid, R42 is a hydro-phobic amino acid, R43 is a neutral hydrophilic amino acid or a hydrophobic amino acid, and (b) an antibiotic which inhibits DNA gyrase; and (c) an acceptable pharmaceutical carrier, wherein said components (a) and (b) are present in a combined amount effective to inhibit growth of a microbe in a host.

42. The composition of claim 41 wherein the peptide is a magainin peptide.

43. The composition of claim 42 wherein said magainin peptide includes the following basic peptide structure;

-R11-R11-R12-R13-R11-R14-R12-R11-R14-R11-R12-R11-RR11-R11-R14a-(R15)n-R14a-R14-, wherein R11 is a hydrophobic amino acid, R12 is a basic hydrophilic amino acid, R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid, R14 and R14a are hydrophobic or basic hydrophilic amino acids, R15 is glutamic acid or aspartic acid, a hydrophobic amino acid, or a basic hydrophilic amino acid, and n is 0 or 1.

44. The composition of claim 42 wherein said magainin peptide includes the following basic peptide structure, -R14-R11-R14a-R12-R11-R11-R12-R13-R11-R14-R12-R11--R11-R12-, wherein R11 is a hydrophobic amino acid, R12 is a basic hydrophilic amino acid, R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid, and R14 and R14a are hydrophobic or basic hydrophilic amino acids.

45. The use of claim 1 wherein the microbe is a bacterium.

46. The use of claim 1 wherein said at least one biologically active peptide or protein is administered topically.

47. The use of claim 1 wherein said at least one biologically active peptide or protein is administered systemically.

48. The use of claim 1 wherein said antibiotic which inhibits DNA gyrase is administered topically.

49. The use of claim 1 wherein said antibiotic which inhibits DNA gyrase is administered systemically.

50. The use of claim 1 wherein said at least one biologically active peptide or protein and said antibiotic which inhibits DNA gyrate are administered topically.

51. The use of claim 1 wherein said at least one biologically active peptide or protein and said antibiotic which inhibits DNA gyrase are administered systemically.

52. The use of claim 37 wherein the microbe is a bacterium.

53. The use of claim 37 wherein said antibiotic which inhibits DNA gyrase is a quinolone antibiotic.

54. The process of claim 53 wherein said quinolone antibiotic is ciprofloxacin.

55. The process of claim 37 wherein said antibiotic which inhibits DNA gyrase is nalidixic acid.

56. The process of claim 37 wherein said antibiotic which inhibits DNA gyrase is oxolinic acid.

57. The process of claim 37 wherein said antibiotic which inhibits DNA gyrase is cinoxacin.

58. The composition of claim 41 wherein said antibiotic which inhibits DNA gyrase is a quinolone antibiotic.

59. The composition of claim 58 wherein said quinolone antibiotic is ciprofloxacin.

60. The composition of claim 41 wherein said antibiotic which inhibits DNA gyrase is nalidixic acid.

61. The composition of claim 41 wherein said antibiotic which inhibits DNA gyrase is oxolinic acid.

62. The composition of claim 41 wherein said antibiotic which inhibits DNA gyrase is cinoxacin.