Peptide compositions and uses therefor

This invention relates to biologically active peptides, and more 
particularly to novel biologically active peptides and uses therefor. 
In accordance with an aspect of the present invention, there is provided a 
biologically active amphiphilic peptide which includes the following 
structural formula X.sub.a : R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 
--R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 
--R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2, wherein 
R.sub.1 is a basic hydrophilic amino acid, R.sub.2 is a hydrophobic amino 
acid, and R.sub.3 is a neutral hydrophilic or hydrophobic amino acid. 
In one embodiment, the peptide has the structure Y.sub.a --X.sub.a, wherein 
X.sub.a is as hereinabove described and Y.sub.a is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.2 ; or 
(iii) R.sub.2 --R.sub.2 --R.sub.2. 
In accordance with another aspect of the present invention, there is 
provided a biologically active amphiphilic peptide which includes the 
following structural formula X.sub.b : R.sub.1 --R.sub.2 --R.sub.2 
--R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 
--R.sub.3 --R.sub.1 -R.sub.2 --R.sub.2 --R.sub.2, wherein R.sub.1, 
R.sub.2, and R.sub.3 are as hereinabove described. 
In one embodiment, the peptide has the structure X.sub.b --Z.sub.b, wherein 
X.sub.b is the peptide structure hereinabove described and Z.sub.b is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.1 ; or 
(iii) R.sub.2 --R.sub.1 --R.sub.2. 
In accordance with another aspect of the present invention, there is 
provided a biologically active amphiphilic peptide which includes one of 
the following basic structures X.sub.1 through X.sub.7 wherein: 
X.sub.1 is --[R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 
--R.sub.2 ]--.sub.n 
X.sub.2 is --[R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 
--R.sub.1 ]--.sub.n ; 
X.sub.3 is --[R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 
--R.sub.2 ]--.sub.n ; 
X.sub.4 is --[R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 
--R.sub.2 ]--.sub.n ; 
X.sub.5 is --[R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 
--R.sub.3 ]--.sub.n ; 
X.sub.6 is --[R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 
--R.sub.1 ]--.sub.n ; and 
X.sub.7 is --[R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 
--R.sub.2 ]--.sub.n ; 
wherein R.sub.1 is a basic hydrophilic amino acid, R.sub.2 is a hydrophobic 
amino acid, R.sub.3 is a neutral hydrophilic or hydrophobic amino acid, 
and n is from 3 to 5. 
The basic hydrophilic amino acids include, but are not limited to Lys, Arg, 
His, Orn, homoarginine (Har), 2, 4-diamino butyric acid (Dbu), and 
p-aminophenylalanine. 
The hydrophobic amino acids include, but are not limited to Ala, Cys, Phe, 
Gly, Ile, Leu, Met, Pro, Val, Trp, Tyr, norleucine (Nle), norvaline (Nva), 
and cyclohexylalanine (Cha). 
The neutral hydrophilic amino acids include, but are not limited to Asn, 
Gln, Ser and Thr. 
In accordance with one embodiment, when the peptide includes the structure 
X1, the peptide may include the following structure: 
Y.sub.1 --X.sub.1, wherein X.sub.1 is as hereinabove described, and Y is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.2 ; 
(iii) R.sub.1 --R.sub.2 --R.sub.2 ; 
(iv) R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 ; 
(v) R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 ; or 
(vi) R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2, wherein 
R.sub.1, R.sub.2, and R.sub.3 are as hereinabove described 
In accordance with another embodiment, when the peptide includes the 
structure X.sub.1, the peptide may include the following structure: 
x.sub.1 --y.sub.1, wherein x.sub.1 is as hereinabove described, and y.sub.1 
is: 
(i) R.sub.1 ; 
(ii) R.sub.1 --R.sub.2 ; 
(iii) R.sub.1 --R.sub.2 --R.sub.2 ; 
(iv) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3; 
(v) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 ; or 
(vi) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2. 
In accordance with yet another embodiment, the peptide may include the 
following structure: 
(Y.sub.1).sub.a --X.sub.1 --(Z.sub.1).sub.b, wherein Y.sub.1 and Z.sub.1 
are as previously defined, a is 0 or 1, and b is 0 or 1. 
When the peptide includes the structure X.sub.2, the peptide may include 
the following structure: 
Y.sub.2 --X.sub.2, wherein X.sub.2 is as hereinabove described, and Y.sub.2 
is: 
(i) R.sub.1 ; 
(ii) R.sub.2 --R.sub.1; 
(iii) R.sub.2 --R.sub.2 --R.sub.1; 
(iv) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1; 
(v) R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 ; or 
(vi) R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 . 
In another embodiment, the peptide may include the following structure: 
X.sub.2 - Z.sub.2 wherein X.sub.2 is as hereinabove described, and Z.sub.2 
is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.2 ; 
(iii) R.sub.2 --R.sub.2 --R.sub.3 ; 
(iv) R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 ; 
(v) R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 ; or 
(vi) R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 . 
In accordance with yet another embodiment, the peptide may include the 
following structure: 
(Y.sub.2).sub.a --X.sub.2 --( Z.sub.2 ).sub.b, wherein Y.sub.2 and Z.sub.2 
are as previously defined, a is 0 or 1, and b is 0 or 1. 
In accordance with another embodiment, when the peptide includes the 
structure X.sub.3, the peptide may include the following structure: 
Y.sub.3 --X.sub.3 wherein X.sub.3 is as hereinabove described, and Y.sub.3 
is: 
(i) R.sub.2 ; 
(ii) R.sub.1 --R.sub.2 ; 
(iii) R.sub.2 --R.sub.1 --R.sub.2 ; 
(iv) R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 ; 
(v) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 ; or 
(vi) R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2, wherein 
R.sub.1, R.sub.2, and R.sub.3 are as hereinabove described. 
In accordance with another embodiment, when the peptide includes the 
structure X.sub.3, the peptide may include the following structure: 
X.sub.3 --Z.sub.3 wherein X.sub.3 is as hereinabove described, and Z.sub.3 
is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.3 ; 
(iii) R.sub.2 --R.sub.3 --R.sub.1 ; 
(iv) R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 ; 
(v) R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 ; or 
(vi) R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1. 
In accordance with yet another embodiment, the peptide may include the 
following structure: 
(Y.sub.3).sub.a --X.sub.3 --(Z.sub.3).sub.b, wherein Y and Z are as 
previously defined, a is 0 or 1, and b is 0 or 1. 
In accordance with yet another embodiment, when the peptide includes the 
structure X.sub.4, the peptide may include the following structure: 
Y.sub.4 --X.sub.4, wherein X.sub.4 is as hereinabove described, and Y.sub.4 
is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.2 ; 
(iii) R.sub.1 --R.sub.2 --R.sub.2 ; 
(iv) R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 ; 
(v) R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 ; or 
(vi) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2, wherein 
R.sub.1, R.sub.2 and R.sub.3 are as hereinabove described. 
In accordance with another embodiment, when the peptide includes the 
structure X.sub.4, the peptide may include the following structure: 
X.sub.4 --Z.sub.4, wherein X.sub.4 is as hereinabove described, and Z.sub.4 
is: 
(i) R.sub.3 ; 
(ii) R.sub.3 --R.sub.1 ; 
(iii) R.sub.3 --R.sub.1 --R.sub.2 ; 
(iv) R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 ; 
(v) R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 ; or 
(vi) R.sub.3 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2. 
In accordance with yet another embodiment, the peptide may include the 
following structure: 
(Y.sub.4).sub.a --X.sub.4 (Z.sub.4).sub.b, wherein X.sub.4 and Z.sub.4 are 
as previously defined, a is 0 or 1, and b is 0 or 1. 
In accordance with a further embodiment, when the peptide includes the 
structure X.sub.5, the peptide may include the following structure: 
Y.sub.5 --X.sub.5, wherein X.sub.5 is as hereinabove described, and Y.sub.5 
is: 
(i) R.sub.3 ; 
(ii) R.sub.2 --R.sub.3 ; 
(iii) R.sub.2 --R.sub.2 --R.sub.3 ; 
(iv) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 ; 
(v) R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 ; or 
(vi) R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3, wherein 
R.sub.1, R.sub.2, and R.sub.3 are as hereinabove described. 
In accordance with another embodiment, when the peptide includes structure 
X.sub.5, the peptide may include the following structure: 
X.sub.5 --Z.sub.5 wherein X.sub.5 is as hereinabove described, and Z.sub.5 
is: 
(i) R.sub.1 ; 
(ii) R.sub.1 --R.sub.2 ; 
(iii) R.sub.1 --R.sub.2 --R.sub.2 ; 
(iv) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 ; 
(v) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 ; or 
(vi) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 . 
In accordance with yet another embodiment, the peptide may include the 
following structure: 
(Y.sub.5).sub.a --X.sub.5 (Z.sub.5).sub.b, wherein X.sub.5 and Z.sub.5 are 
as previously defined, a is 0 or 1, and b is 0 or 1. 
In accordance with a further embodiment, when the peptide includes the 
structure X.sub.6, the peptide may include the following structure: 
Y.sub.6 --X.sub.6 wherein X.sub.6 is as hereinabove described, and Y.sub.6 
is: 
(i) R.sub.1 ; 
(ii) R.sub.3 --R.sub.1 ; 
(iii) R.sub.2 --R.sub.3 --R.sub.1 ; 
(iv) R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 ; 
(v) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 ; or 
(vi) R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1, wherein 
R.sub.1, R.sub.2, and R.sub.3 are as hereinabove described. 
In accordance with another embodiment, when the peptide includes the 
structure X.sub.6, the peptide may include the following structure: 
X.sub.6 --Z.sub.6, wherein X.sub.6 is as hereinabove described, and Z.sub.6 
is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.2 ; 
(iii) R.sub.2 --R.sub.2 --R.sub.1 ; 
(iv) R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 ; 
(v) R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 ; or 
(vi) R.sub.2 --R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3. 
In accordance with yet another embodiment, the peptide may include the 
following structure: 
(Y.sub.6).sub.a --X.sub.6 (Z6).sub.b, wherein Y.sub.6 and Z.sub.6 are as 
previously defined, a is 0 or 1, and b is 0 or 1. 
In accordance with one embodiment, when the peptide includes the structure 
X.sub.7, the peptide may include the structure Y.sub.7 --X.sub.7, wherein 
X.sub.7 is as hereinabove described, and Y.sub.7 is: 
(i) R.sub.2 ; 
(ii) R.sub.1 --R.sub.2 ; 
(iii) R.sub.3 --R.sub.1 --R.sub.2 ; 
(iv) R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 ; 
(v) R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2 ; or 
(vi) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 --R.sub.2, wherein 
R.sub.1, R.sub.2, and R.sub.3 are as hereinabove described. 
In accordance with a further embodiment, when the peptide includes the 
structure X.sub.7, the peptide may include the following structure: 
X.sub.7 --Z.sub.7 wherein X.sub.7 is as hereinabove described, and Z.sub.7 
is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.1 ; 
(iii) R.sub.2 --R.sub.1 --R.sub.2 ; 
(iv) R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 ; 
(v) R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 ; or 
(vi) R.sub.2 --R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.1 . 
In accordance with yet another embodiment, the peptide may include the 
following structure: 
(Y.sub.7).sub.a --X.sub.7 (Z.sub.7).sub.b, wherein Y.sub.7 and Z.sub.7 are 
as previously defined, a is 0 or 1, and b is 0 or 1. 
The peptides and/or analogives or derivatives thereof, may be C-terminal 
acids or amides. 
In a preferred embodiment, n is 3, and most preferably the peptide is of 
one of the following structures listed below and also listed in the 
accompanying sequence listing: 
(Lys Ile Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 1) 
(Lys Ile Ala Lys Ile Ala Gly).sub.3 --NH.sub.2 (SEQ ID NO: 2) 
(Lys Ile Ala Gly Lys Ile Gly).sub.3 --NH.sub.2 (SEQ ID NO: 3) 
(Lys Leu Ala Gly Lys Leu Ala).sub.3 --NH.sub.2 (SEQ ID NO: 4) 
(Lys Phe Ala Gly Lys Phe Ala).sub.3 --NH.sub.2 (SEQ ID NO: 5) 
(Lys Ala Leu Ser Lys Ala Leu).sub.3 --NH.sub.2 (SEQ ID NO: 6) 
(Lys Leu Leu Lys Ala Leu Gly).sub.3 --NH.sub.2 (SEQ ID NO: 7) 
(Lys Ala Ile Gly Lys Ala Ile).sub.3 --NH.sub.2 (SEQ ID NO: 8) 
(Gly Ile Ala Lys Ile Ala Lys).sub.3 --NH.sub.2 (SEQ ID NO: 9) 
(Lys Ile Ala Lys Ile Phe Gly).sub.3 --NH.sub.2 (SEQ ID NO: 10) 
(Gly Ile Ala Arg Ile Ala Lys).sub.3 --NH.sub.2 (SEQ ID NO: 11) 
(Lys Phe Ala Arg Ile Ala Gly).sub.3 --NH.sub.2 (SEQ ID NO: 12) 
(Gly Phe Ala Lys Ile Ala Lys).sub.3 --NH.sub.2 (SEQ ID NO: 13) 
(Lys Ile Ala Gly Orn Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 14) 
(Lys Ile Ala Arg Ile Ala Gly).sub.3 --NH.sub.2 (SEQ ID NO: 15) 
(Orn Ile Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 16) 
(Gly Ile Ala Arg Ile Phe Lys).sub.3 --NH.sub.2 (SEQ ID NO: 17) 
(Lys Nle Ala Gly Lys Nle Ala).sub.3 --NH.sub.2 (SEQ ID NO: 18) 
(Lys Nle Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 19) 
(Lys Ile Ala Gly Lys Nle Ala).sub.3 --NH.sub.2 (SEQ ID NO: 20) 
(Lys Nva Ala Gly Lys Nva Ala).sub.3 --NH.sub.2 (SEQ ID NO: 21) 
(Lys Nva Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 22) 
(Lys Leu Leu Ser Lys Leu Gly).sub.3 --NH.sub.2 (SEQ ID NO: 23) 
(Lys Leu Leu Ser Lys Phe Gly).sub.3 --NH.sub.2 (SEQ ID NO: 24) 
(Lys Ile Ala Gly Lys Nva Ala).sub.3 --NH.sub.2 (SEQ ID NO: 25) 
(His Ile Ala Gly His Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 26) 
(Ala Gly Lys Ile Ala Lys Ile).sub.3 --NH.sub.2 (SEQ ID NO: 27) 
(Ile Ala Lys Ile Ala Gly Lys).sub.3 --NH.sub.2 (SEQ ID NO: 28) 
(Lys Ile Ala Gly Arg Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 29) 
(Arg Ile Ala Gly Arg Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 30) 
(Lys Val Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO: 31) 
(Lys Ile Ala Gly Lys Val Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 32) 
(Ala Lys Ile Ala Gly Lys Ile).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 33) 
(Orn Ile Ala Gly Orn Ile Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 34) 
(Lys Phe Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 35) 
(Lys Ile Ala Gly Lys Phe Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 36) 
(Lys Cha Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 37) 
(Lys Nle Ala Lys Ile Ala Gly).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 38) 
(Arg Ile Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 39) 
(Har Ile Ala Gly Har Ile Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 40) 
(Xaa Ile Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 41) 
(Lys Ile Ala Gly Xaa Ile Ala).sub.3 --NH.sub.2 (SE.sub.Q ID NO: 42) 
Lys Ile Ala (Lys Ile Ala Gly Lys Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO:43) 
In (SEQ ID NO:41) and (SEQ ID NO:42), Xaa is p-aminophenylalanine. 
In accordance with another aspect of the present invention, there is 
provided a biologically active amphiphilic peptide which includes the 
following basic structure X.sub.14 : 
EQU R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.4 --R.sub.2 --R.sub.2 
--R.sub.1 --R.sub.2 --R.sub.2 --R.sub.2 --R.sub.4 --R.sub.2 --R.sub.2, 
wherein R.sub.1, R.sub.2, and R.sub.3 are as hereinabove described, and 
R.sub.4 is a basic hydrophilic or hydrophobic amino acid. 
In accordance with one embodiment, the peptide may include the following 
structure: 
Y.sub.14 --X.sub.14, wherein X.sub.14 is as hereinabove described, and 
Y.sub.14 is: 
(i) R.sub.2 ; 
(ii) R.sub.2 --R.sub.2 ; 
(iii) R.sub.4 --R.sub.2 --R.sub.2 ; 
(iv) R.sub.3 --R.sub.4 --R.sub.2 --R.sub.2 ; 
(v) R.sub.2 --R.sub.3 --R.sub.4 --R.sub.2 --R.sub.2 ; 
(vi) R.sub.2 --R.sub.2 --R.sub.3 --R.sub.4 --R.sub.2 --R.sub.2, or 
(vii) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.4 --R.sub.2 --R.sub.2, 
wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are as hereinabove 
described. 
In accordance with another embodiment, the peptide may include the 
following structure: 
X.sub.14 --Z.sub.14, wherein X.sub.14 is as hereinabove described and 
Z.sub.14 is: 
(i) R.sub.1 ; 
(ii) R.sub.1 --R.sub.2 ; 
(iii) R.sub.1 --R.sub.2 --R.sub.2 ; 
(iv) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 ; 
(v) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.4 ; 
(vi) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.4 --R.sub.2 ; or 
(vii) R.sub.1 --R.sub.2 --R.sub.2 --R.sub.3 --R.sub.4 --R.sub.2 --R.sub.2, 
wherein R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are as hereinabove 
described. 
In accordance with yet another embodiment the peptide may include the 
following structure: 
(Y.sub.14).sub.a --X.sub.14 --(Z.sub.14).sub.b, wherein X and Y are as 
previously defined, a is 0 or 1, and b is 0 or 1. In a preferred 
embodiment, the peptide has the following structural formula as indicated 
in the accompanying sequence listing: 
(SEQ ID NO: 44)--NH.sub.2. 
In another preferred embodiment, the peptide has the following structural 
formula as indicated in the accompanying sequence listing: 
(SEQ ID NO: 45)--NH.sub.2. 
In accordance with a further embodiment, the peptide has one of the 
following structural formulae as indicated in the accompanying sequence 
listing: 
(SEQ ID NO: 46)--NH.sub.2 
(SEQ ID NO: 47)--NH.sub.2 
(SEQ ID NO: 48)--NH.sub.2 
(SEQ ID NO: 49)--NH.sub.2 
(SEQ ID NO: 50)--NH.sub.2 
(SEQ ID NO: 51)--NH.sub.2 
(SEQ ID NO: 52)--NH.sub.2 
(SEQ ID NO: 53)--NH.sub.2 
(SEQ ID NO: 54)--NH.sub.2 
(SEQ ID NO: 55)--NH.sub.2 
(SEQ ID NO: 56)--NH.sub.2 
(SEQ ID NO: 57)--NH.sub.2 
(SEQ ID NO: 58)--NH.sub.2 
(SEQ ID NO: 59)--NH.sub.2 
(SEQ ID NO: 60)--NH.sub.2 
(SEQ ID NO: 61)--NH.sub.2 
In accordance with another aspect of the present invention, there is 
provided a biologically active amphiphilic peptide which includes the 
following structural formula: 
--(Lys Ile Ala Lys Lys Ile Ala)--.sub.n wherein n is from 2 to 5. 
Preferably, n is 3, and the peptide has the following structural formula: 
(Lys Ile Ala Lys Lys Ile Ala).sub.3 --NH.sub.2. (SEQ ID NO:62). 
In accordance with another aspect of the present invention, there is 
provided a biologically active amphiphilic peptide which includes the 
following structural formula: 
--(Lys Phe Ala Lys Lys Phe Ala).sub.n --, wherein n is from 2 to 5. 
Preferably, n is 3, and the peptide has the following structural formula: 
(Lys Phe Ala Lys Lys Phe Ala).sub.3 --NH.sub.2 (SEQ ID NO:63). 
In accordance with yet another aspect of the present invention, there is 
provided a biologically active amphiphilic peptide which includes the 
following structural formula: 
--(Lys Phe Ala Lys Lys Ile Ala).sub.n --, wherein n is from 2 to 5. 
Preferably, n is 3, and the peptide has the following structural formula: 
(Lys Phe Ala Lys Lys Ile Ala).sub.3 --NH.sub.2 (SEQ ID NO:64). 
In accordance with another aspect of the present invention, there is 
provided a biologically active amphiphilic peptide selected from the group 
consisting of the following structural formulae as given in the 
accompanying sequence listing: 
(SEQ ID NO: 65)--NH.sub.2 
(SEQ ID NO: 66)--NH.sub.2 
(SEQ ID NO: 67)--NH.sub.2 
(SEQ ID NO: 68)--NH.sub.2 
In accordance with one embodiment, each of the amino acid residues 
contained in the peptides is a D-amino acid residue or glycine. Although 
the scope of this particular embodiment is not to be limited to any 
theoretical reasoning, it is believed that the above-mentioned peptides, 
when consisting entirely of D-amino acid or glycine residues, may have 
increased resistance to proteolytic enzymes while retaining their 
biological activity. Such peptides thus may be administered orally. Thus 
in accordance with a preferred embodiment, all of the amino acid residues 
are either D-amino acid or glycine residues or L-amino acid or glycine 
residues. 
An amphiphilic peptide is a peptide which includes both hydrophobic and 
hydrophilic peptide regions. 
In general, the peptides hereinabove described, and/or analogues or 
derivatives thereof are generally water soluble to a concentration of at 
least 20 mg/ml at neutral pH in water. Such peptides are capable of 
forming an.alpha.-helical structure. In addition, the structure of such 
peptide provides for flexibility of the peptide molecule. When the peptide 
is placed in water, it does not assume an amphiphilic structure. When the 
peptide encounters an oily surface or membrane, the peptide chain folds 
upon itself into a rod-like structure. 
In general, the peptides of the present invention are ion channel-forming 
peptides. An ion channel-forming peptide or ionophore is a peptide which 
increases the permeability for ions across a natural or synthetic lipid 
membrane. B. Christensen, et al., PNAS, Vol. 85, pgs. 5072-5076 (July 
1988) describes methodology which indicates whether or not a peptide has 
ion channel-forming properties and is therefore an ionophore. As used 
herein an ion channel-forming peptide is a peptide which has ion 
channel-forming properties as determined by the method of Christensen, et 
al. 
The peptides and/or analogues or derivatives thereof may be administered to 
a host; for example a human or non-human animal, in an amount effective to 
inhibit growth of a target cell, virus, or vitally-infected cell. Thus, 
for example, the peptides and/or analogues or derivatives thereof may be 
used as anti-microbial agents, anti-viral agents, antibiotics, anti-tumor 
agents, antiparasitic agents, antifungal agents, spermicides, as well as 
exhibiting other bioactive functions. 
The term "antimicrobial" as used herein means that the peptides of the 
present invention inhibit, prevent, or destroy the growth or proliferation 
of microbes such as bacteria, fungi, or the like. 
The term "antibiotic" as used herein means that the peptides employed in 
the present invention produce effects adverse to the normal biological 
functions of the non-host cell, tissue or organism, including death or 
destruction and prevention of the growth or proliferation of the non-host 
cell, tissue, or organism, when contacted with the peptides. 
The term "spermicidal" as used herein means that the peptides employed in 
the present invention, inhibit, prevent, or destroy the motility of sperm. 
The term "antiviral" as used herein means that the peptides employed in the 
present invention inhibit, prevent, or destroy the growth or proliferation 
of viruses, or of virally-infected cells. 
The term anti-tumor as used herein means that the peptide inhibits the 
growth of or destroys tumors. 
The term "antifungal" as used herein means that the peptides of the present 
invention may be used to inhibit the growth of or destroy fungi. 
The term "antiparasitic" as used herein means that the peptides of the 
present invention may be used to inhibit the growth of or destroy 
parasites. 
The peptides may be administered in vivo or in vitro. The peptides also may 
be administered directly to a target cell, virus, or virally-infected 
cell, or the peptides may be administered systemically. 
The peptides of the present invention have a broad range of potent 
antibiotic activity against a plurality of microorganisms including 
Gram-positive and Gram-negative bacteria, fungi, protozoa, and the like, 
as well as parasites. The peptides of the present invention allow a method 
for treating or controlling microbial infection caused by organisms which 
are sensitive to the peptides. Such treatment may comprise administering 
to a host organism or tissue susceptible to or affiliated with a microbial 
infection an antimicrobial amount of at least one of the peptides. 
Because of the antibiotic, antimicrobial, and antiviral properties of the 
peptides, they may also be used as preservatives or sterilants of 
materials susceptible to microbial or viral contamination. 
The peptide and/or derivatives or analogues thereof may be administered in 
combination with a non-toxic pharmaceutical carrier or vehicle such as a 
filler, non-toxic buffer, or physiological saline solution. Such 
pharmaceutical compositions may be used topically or systemically and may 
be in any suitable form such as a liquid, solid, semi-solid, injectable 
solution, tablet, ointment, lotion, paste, capsule, or the like. The 
peptide compositions may also be used in combination with adjuvants, 
protease inhibitors, or compatible drugs where such a combination is seen 
to be desirable or advantageous in controlling infection caused by harmful 
microorganisms including protozoa, viruses, and the like, as well as by 
parasites. 
The peptide(s) of the present invention may be administered to a host; in 
particular an animal, in an effective antibiotic and/or anti-tumor and/or 
anti-viral and/or anti-microbial and/or anti-parasitic and/or an 
antispermicidal amount. 
Depending on the use, a composition in accordance with the invention will 
contain an effective anti-microbial amount and/or an effective 
antispermicidal amount and/or an effective anti-viral amount and/or an 
effective anti-tumor amount and/or an effective antibiotic amount and/or 
anti-parasitic amount of one or more of the hereinabove described peptides 
which have such activity. 
The peptide of the present invention may also be employed in promoting or 
stimulating healing of a wound in a host. 
The term "wound healing" as used herein includes various aspects of the 
would healing process. 
These aspects include, but are not limited to, increased contraction of the 
wound, increased deposition of connective tissue, as evidenced by, for 
example, increased deposition of collagen in the wound, and increased 
tensile strength of the wound, i.e., the peptides increase wound breaking 
strength. The peptides of the present invention may also be employed so as 
to reverse the inhibition of wound healing caused by steroids such as 
cortisone or by conditions which compromise or depress the immune system. 
The peptides of the present invention may be used in the treatment of 
external burns and to treat and/or prevent skin and burn infections. In 
particular, the peptides may be used to treat skin and burn infections 
caused by organisms such as, but not limited to, P. aeruginosa and S. 
aureus. 
The peptides are also useful in the prevention or treatment of eye 
infections. Such infections may be caused by bacteria such as, but not 
limited to, P. aeruginosa, S. aureus, and N. gonorrhoeae, by fungi such as 
but not limited to C. albicans and A. fumigatus, by parasites such as but 
not limited to A. castellani, or by viruses. 
The peptides may also be effective in killing cysts, spores, or 
trophozoites of infection--causing organisms. Such organisms include, but 
are not limited to Acanthamoeba which forms trophozoites or cysts, C. 
albicans, which forms spores, and A. fumigatus, which forms spores as 
well. 
The peptides may also be administered to plants in an effective 
antimicrobial or antiviral or antiparasitic amount to prevent or treat 
microbial or viral or parasitic contamination thereof. 
In general, the peptide is employed to provide peptide dosages of from 0.1 
mg. to 500 mg. per kilogram of host weight, when administered 
systemically. When administered topically, the peptide is used in a 
concentration of from 0.05% to 10%. 
The peptides may be produced by known techniques and obtained in 
substantially pure form. For example, the peptides may be synthesized on 
an automatic peptide synthesizer. Journal of the American Chemical 
Society, Vol. 85, pgs. 2149-54 (1963). It is also possible to produce such 
peptides by genetic engineering techniques. The codons encoding the amino 
acids are known to those skilled in the art, and thus one may construct 
DNA encoding any of the peptides by accepted techniques, and clone such 
DNA into an expression vehicle such as, for example, a plasmid, and 
transfect such an expression vehicle into a cell which will express the 
peptide. Thus, it is contemplated within the scope of the present 
invention that one may administer the peptide to a host by administering 
to a host DNA encoding the peptides. 
In accordance with another embodiment, the peptides of the present 
invention may be employed in combination with an ion having 
pharmacological properties for the purposes hereinabove described. 
An ion having pharmacological properties is one which when introduced into 
a target cell, virus, or virally-infected call, inhibits and/or prevents 
and/or destroys the growth of the target cell, virus, or virally-infected 
cell. 
Such an ion having pharmacological properties is one which in the absence 
of an ion channel forming peptide is unable to cross a natural or 
synthetic lipid membrane; in particular a cell membrane, in sufficient 
amounts to affect a cell adversely. 
The peptide and ion having pharmacological properties may be administered 
as a single composition or in separate compositions, and the single or 
separate compositions may include additional materials, actives and/or 
inactives, in addition to the peptide and ion having pharmacological 
properties. As representative examples of ions having pharmacological 
properties which may be employed, there may be mentioned fluoride, 
peroxide, bicarbonate, and silver ions. 
The peptide and the ion having pharamacological properties, whether 
administered or prepared in a single composition or in separate 
compositions, are employed in amounts effective to inhibit and/or prevent 
and/or destroy the growth of the target cell, virus, or virally-infected 
cell. In effect, the ion potentiates the action of the peptide, i.e., the 
amount of ion is effective to reduce the minimum effective concentration 
of the peptide for inhibiting growth of a target cell, virus, or 
virally-infected cell. 
The ion having pharamcological properties when used topically, is generally 
employed in a concentration of from 0.05% to 2.0%. When used systemically, 
the ion is generally employed in an amount of from 1 to 10 mg. per kg. of 
host weight. Peptide dosages may be within the ranges hereinabove 
described. 
It is also to be understood that the peptide and ion having 
pharamacological properties may be delivered or administered in different 
forms; for example, the ion having pharamacological properties may be 
administered orally, while the peptide may be administered by IV or IP. 
As representative examples of administering the peptide and ion having 
pharamcological properties for topical or local administration, the 
peptide could be administered in an amount of up to about 1% weight to 
weight and the ion having pharamacological properties delivered in an 
amount of about 50 mM (about 0.1%). Alternatively, the ion, in the form of 
a salt such as sodium fluoride, could be administered orally in 
conjunction with systemic administration of the peptide. For example, the 
peptide may be administered IV or IP to achieve a serum dose of 100 
micrograms per milliliter (10 milligrams per kilogram) in conjunction with 
an oral dose of ion, in particular, sodium fluoride, of 10 meq per 
kilogram. 
In accordance with another embodiment, the peptides of the present 
invention may be administered to a host in combination with an antibiotic 
selected from the class consisting of bacitracins, gramacidin, polymyxin, 
vancomycin, teichoplanin, aminoglycosides, hydrophobic antibiotics, 
penicillins, monobactams, or derivatives or analogues thereof. 
The bacitracins, gramacidin, polymyxin, vancomycin, teichoplanin, and 
derivatives and analogues thereof, are a group of polypeptide antibiotics. 
A preferred bacitracin is bacitracin A. 
Aminoglycoside antibiotics include tobramycin, kanamycin, amikacin, the 
gentamicins (e.g., gentamicin C.sub.1, gentamicin C.sub.2, gentamicin 
C.sub.1a), netilmicin, kanamycin, and derivatives and analogues thereof. 
The preferred aminoglycosides are tobramycin and the gentamicins. The 
aminoglycosides, and the bacitracins hereinabove described, tend to be 
hydrophilic and water-soluble. 
Penicillins which may be employed include, but are not limited to benzyl 
penicillin, ampicillin, methicillin (dimethoxyphenyl penicillin), 
ticaricillin, penicillin V (phenoxymethyl penicillin), oxacillin, 
cloxacillin, dicloxacillin, flucloxacillin, amoxicillin, and 
amidinocillin. Preferred penicillins which may be employed are benzyl 
penicillin and ampicillin. A preferred monobactam which may be employed is 
aztreonam. 
As representative examples of hydrophobic antibiotics which may be used in 
the present invention, there may be mentioned macrolides such as 
erythromycin, roxythromycin, clarithromycin, etc.; 9-N-alkyl derivatives 
of erythromycin; midecamycin acetate; azithromycin; flurithromycin; 
rifabutin; rokitamycin; a 6-O-methyl erythromycin A known as TE-031 
(Taisho); rifapentine; benzypiperazinyl rifamycins such as CGP-7040, 
CGP-5909, CGP-279353 (Ciba-Geigy); an erythromycin A derivative with a 
cyclic carbamate fused to the C.sub.11 /C.sub.12 position of a macrolide 
ring known as A-62514 (Abbott); AC-7230 (Toyo Jozo); benzoxazinorifamycin; 
difficidin; dirithromycin; a 3-N-piperdinomethylzaino methyl rifamycin SV 
known as FCE-22250 (Farmitalia); M-119-a (Kirin Brewery); a 
6-O-methyl-1-4"-O-carbamoyl erythromycin known as A-63075 (Abbott); 
3-formylrifamycin SV-hydrazones with diazabicycloalkyl side chains such as 
CGP-27557 and CGP-2986 (Ciba-Geigy); and 16-membered macrolides having a 
3-O-alpha-L-cladinosyl moiety, such as 3-O-alpha-L-cladinosyldeepoxy 
rosaramicin; tylosins and acyl demycinosyl tylosins. 
In addition to the macrolides hereinabove described, rifamycin, 
carbenicillin, and nafcillin may be employed as well. 
Other antibiotics which may be used (whether or not hydrophobic) are 
antibiotics which are 50-S ribosome inhibitors such as lincomycin; 
clindamycin; and chloramphenicol; etc.; antibiotics which have a large 
lipid like lactone ring, such as mystatin; pimaricin, etc. 
The peptide and antibiotic may be adminstered by direct administration to a 
target cell or by systemic or topical administration to a host which 
includes the target cell, in order to prevent, destroy or inhibit the 
growth of a target cell. Target cells whose growth may be prevented, 
inhibited, or destroyed by the administration of the peptides and 
antibiotic include Gram-positive and Gram-negative bacteria as well as 
fungal cells. 
The antibiotic, such as those hereinabove described, or derivatives or 
analogues thereof, when used topically, is generally employed in a 
concentration of about 0.1% to about 10%. When used systemically, the 
antibiotic or derivative or analogue thereof is generally employed in an 
amount of from 1.25 mg. to about 45 mg. per kg. of host weight per day. 
Peptide dosages may be those as hereinabove described. 
As representative examples of administering the peptide and antibiotic for 
topical or local administration, the peptide could be administered in an 
amount of from about 0.1% to about weight to weight, and the antibiotic is 
delivered in an amount of from about 0.1% to about 10% weight to weight. 
In accordance with another embodiment, the peptides of the present 
invention may be administered in combination with an antiparasitic agent 
or an antifungal agent. 
Antiparasitic agents which may be employed include, but are not limited to, 
anti-protozoan agents. Examples of specific anti-parasitic agents which 
may be employed include, but are not limited to, pentamidine isethionate, 
and propamidine isethionate (Brolene). 
Anti-fungal agents which may be employed include, but are not limited to, 
ketoconazole. It is also to be understood that certain anti-parasitic 
agents may also have anti-fungal activity, and that certain anti-fungal 
agents may have anti-parasitic activity. 
In accordance with another embodiment, the peptides of the present 
invention may be administered in combination with an antibiotic which 
inhibits DNA gyrase, which is an enzyme involved in the formation of bonds 
between individual coiling strands of replicating bacterial DNA. Thus, DNA 
gyrase is necessary for the normal replication of bacterial DNA, and, 
therefore, antibiotics which inhibit DNA gyrase inhibit the normal 
replication of bacterial DNA. 
Examples of antibiotics which inhibit DNA gyrase include nalidixic acid, 
oxolinic acid, cinoxacin, and quinolone antibiotics which include 
ciprofloxacin, norfloxacin, ofloxacin, enoxacin, pefloxacin, lomefloxacin, 
fleroxacin, tosulfloxacin, temafloxacin, and rufloxacin. 
In accordance with another embodiment, the peptides of the present 
invention may be administered for the purpose hereinabove described in 
combination with other biologically active amphiphilic peptides, or in 
combination with ion channel-forming proteins. 
Examples of biologically active amphiphilic peptides which may be employed 
in combination with the peptides of the present invention include magainin 
peptides, PGLa peptides, XPF peptides, CPF peptides, cecropins and 
sarcotoxins. 
A magainin peptide is either a magainin such as magainin I, II, or III or 
an analogue or derivative thereof. 
The magainin peptides generally include at least fourteen amino acids. A 
magainin peptide preferably has 22 or 23 amino acids. 
As representative examples of such magainin peptides, there may be 
mentioned peptides having the following peptide sequences as listed in the 
accompanying sequence listing: 
(a) (NH.sub.2) (SEQ ID. NO: 69)(OH) or (NH.sub.2) (Magainin I) 
(b) (NH.sub.2) (SEQ ID. NO: 70)(OH) or (NH.sub.2) (Magainin II) 
(c) (NH.sub.2) (SEQ ID. NO. 71)(OH) or (NH.sub.2) (Magainin III) 
The following are examples of peptide derivatives or analogues: 
(d) (NH.sub.2) (SEQ ID. NO: 72) (OH) or (NH.sub.2) 
(e) (NH.sub.2) (SEQ ID. NO: 73) (OH) or (NH.sub.2) 
(f) (NH.sub.2) (SEQ ID. NO: 74) (OH) or (NH.sub.2) 
Magainin peptides are described in Proc. Natl. Acad. Sci., Vol. 84, pgs. 
5449-53 (Aug. 1987). The term "magainin peptides" as used herein refers to 
the magainin peptides as well as derivatives and analogues thereof 
including but not limited to the representative derivatives and analogues. 
A PGLa peptide is either PGLa or an analogue or derivative thereof. 
The PGLa peptides generally include at least seventeen amino acids and may 
include as many as forty amino acids. 
An XPF peptide is either XPF or an analogue or derivative thereof. The XPF 
peptides generally include at least nineteen amino acids and may include 
up to forty amino acids. 
As representative examples of PGLa and XPF peptides, there may be mentioned 
the following peptide sequences as well as appropriate analogues and 
derivatives thereof: 
PGLa: (SEQ ID NO: 75) (NH.sub.2) 
XPF: (SEQ ID NO: 76) 
A review of XPF and PGLa can be found in Hoffman, et al., EMBO J., 
2:711-714 (1983); Andreu, et al., J. Biochem, 149:531-535 (1985); Gibson, 
et al., J. Biol. Chem., 261: 5341-5349 (1986); and Giovannini, et al., 
Biochem J., 243:113-120 (1987). 
A CPF peptide is either a CPF peptide or an analogue or derivative thereof. 
In general, a CPF peptide does not include more than 40 amino acids. 
Representative examples of CPF peptides which may be employed in 
combination with the peptides of the present invention, some of which have 
been described in the literature, include the following sequences: 
(I) (SEQ ID NO: 77) 
(II) (SEQ ID NO: 78) 
(III) (SEQ ID NO: 79) 
(IV) (SEQ ID NO: 80) 
(V) ( SEQ ID NO: 81) 
(VI) (SEQ ID NO: 82) 
(VII) (SEQ ID NO: 83) 
(VIII) (SEQ ID NO: 85) 
(IX) (SEQ ID NO: 85) 
(X) (SEQ ID NO: 86) 
(XI) (SEQ ID NO: 87) 
(XII) (SEQ ID NO: 88) 
(XIII) (SEQ ID NO: 89) 
The above is expressed as single letter code for amino acids. 
A review of the CPF peptides can be found in Richter, K., Egger, R., and 
Kreil (1986) J. Biol. Chem. 261, 3676-3680; Wakabayashi, T. Kato, H., and 
Tachibaba, S. (1985) Nucleic Acids Research 13, 1817-1828; Gibson, B. W., 
Poulter, L., Williams, D. H., and Maggio, J. E. (1986) J. Biol. Chem. 261, 
5341-5349. 
The term cecropins includes the basic structure as well as analogues and 
derivatives thereof. The cecropins and analogues and derivatives thereof 
are described in Ann. Rev. Microbiol. 1987, Vol. 41, pages 103-26, in 
particular page 108, and in Christensen, et al., PNAS Vol. 85, pgs. 
5072-76, which are hereby incorporated by reference. 
The term sarcotoxins includes the basic materials as well as analogues and 
derivatives thereof. The sarcotoxins and analogues and derivatives thereof 
are described in Molecular Entomology, pages 369-78, in particular page 
375, Alan R. liss, Inc. (1987), which is hereby incorporated by reference. 
Ion channel-forming proteins or peptides which may be employed in 
combination with the peptides of the present invention include defensins, 
also known as human neutrophil antimicrobial peptides (HNP), major basic 
protein (MBP) of eosinophils, bactericidal permeability-increasing protein 
(BPI), and a pore-forming cytotoxin called variously perforin, cytolysin, 
or pore-forming protein. Defensins are described in Selsted, et al., J. 
Clin. Invest., Vol. 76, pgs. 1436-1439 (1985). 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, pgs. 
14891-14894 (1987). Perforin is 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 incorporated by reference. 
The term ion channel-forming proteins includes the basic structures of the 
ion channel-forming proteins as well as analogues and derivatives.