AU693518B2

AU693518B2 - Ion-channel forming amphiphilic peptides having n-terminal modifications

Info

Publication number: AU693518B2
Application number: AU17288/95A
Authority: AU
Inventors: U. Prasad Kari; Michael Mclane; Taffy J Williams
Original assignee: Magainin Pharmaceuticals Inc
Current assignee: Magainin Pharmaceuticals Inc
Priority date: 1994-01-18
Filing date: 1995-01-18
Publication date: 1998-07-02
Anticipated expiration: 2015-01-18
Also published as: CA2180748A1; JPH09507669A; EP0750632A1; AU1728895A; WO1995019370A1

Description

ION-CHANNEL FORMING AMPHIPHILIC PEPTIDES HAVING N-TERMINAL MODIFICATIONS.

Technical Field This invention relates to biologically active peptides.

More particularly, this invention relates to biologically active peptides having N-terminal (or amino-terminal) substitutions.

In accordance with an aspect of the present invention, there is provided an N-terminal substituted peptide or protein having the formula:

W

1 T N X, wherein X is a biologically active peptide or protein. The peptide or protein is preferably an ion channel-forming peptide or protein. T is a lipophilic moiety, and W is T or hydrogen.

The term "lipophilic," as used herein, means that the lipophilic moiety enhances the interaction of the peptide or o• protein with a lipid membrane, such as, for example, a cell membrane.

Lipophilic moieties which may be employed, include, but are not limited to, any moiety which may be placed on the N-terminal of the peptide through a condensation reaction with nitrogen. The lipophilic moiety T may be, for example, a carboxylic acid, a phosphoric acid, preferably an alkylphosphoric acid, a phosphonic acid, preferably an alkylphosphonic acid, a sulfonic acid, preferably an alkylsulfonic acid, or an alkyl group. Preferably, T is: So• O R -C wherein R is a hydrocarbon having at least two and no more than 16 carbon atoms.

In one embodiment, R is an alkyl group. The alkyl group be a straight chain or branched chain alkyl group; or a I I wn o/1104l7n PCTIJS95/1007 14 2 cycldalkyl group. For example, R may be CH 3

(CH

2 wherein n is from 1 to 14. Preferably, n is from 3 to 12, more preferably from 4 to 11, still more preferably from 6 to 11, and most preferably n is 6, whereby T is an octanoyl group.

In another embodiment, R is an aromatic (including phenyl and naphthyl), or an alkyl aromatic group. For example, R may be O-(CH 2 wherein z is from 0 to 6.

In another embodiment, R is

CH

3 CH 3

I

CH-(CH

2 n-O--CH-

CH

3 wherein n is from 1 to Preferably n is 1, whereby R is*an ibuprofyl group.

In yet another embodiment, T is:

O

I I

HOOC-(CH

2 wherein x is from 1 to 14. Preferably, x is 2, and T is a succinyl group.

In another embodiment, T is:

CH

3

(CH

2 y-CH=CH-CH-CH-NH-, OH wherein y is from 1 to 14.

Preferably, y is 12, whereby T is a sphingosine group.

.In yet another embodiment, T is: O 0 1 II I I CH (CH2 )-CH=CH-CH-CH-NH-C- (CH2 OH wherein x and y are hereinabove described. Preferably, x is 2, and y is 12.

In one embodiment, W is hydrogen.

Applicant has found, that when biologically active peptides have substitutions at the N-terminal such as those SUBSTITUTE SHEET (RULE 26)

I

wn )5/1R70 PrT/U"ncOC7n i ,I 3 hereinabove described, such peptides have increased biological activity against target cells, viruses, and virally-infected cells, as compared with unsubstituted peptides or peptides substituted at the N-terminal with an acetyl group. Applicant also has found that the N-terminal substitutions hereinabove described significantly increase the biological activity of "short" peptides, i.e. peptides having no more than 14 amino acid residues.

As hereinabove stated, the biologically active peptides or proteins of the present invention are preferably ion channel-forming peptides. An ion channel-forming peptide or protein or ionophore is a peptide or protein which increases the permeability for ions across a natural or synthetic lipid membrane. B. Christensen, et al., PNAS, Vol. pgs. 5072-5076 (July 1988) describes methodology which indicates whether or not a peptide or protein 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 channel-forming properties as determined by the method of Christensen, et al.

An amphophilic peptide or protein is a peptide or protein which includes both hydrophobic and hydrophilic peptide or protein regions.

The ion channel-forming peptides employed in the present invention are generally water soluble to a concentration of at least 20 mg/ml at neutral pH in water. In addition, the structure of such peptide provides for flexibility of the peptide molecule. Such peptides are capable of forming an alpha-helical structure. When the peptide is placed in water, it does not assume an amphophilic structure. When the peptide encounters an oily surface or membrane, the peptide chain folds upon itself into a rodlike structure.

In general, such peptides have at least 7 amino acids, and in many cases have at least 20 amino acids. In most cases, such peptides do not have in excess of 40 amino acids.

SUBSTI UTE SHEET (RULE 28)

-B

WO 95/19370 4 PCT/IJS95/00714 The peptides and/or analogues or derivatives thereof may be administered to a host; for example a human or non-human animal, in am amount effective to inhibit growth of a target cell, virus, or virally-infected cell. Thus, for example, the peptides and/or analogues or derivatives thereof may be used as antimicrobial agents, anti-viral agents, antibacterial agents, anti-tumor agents, anti-parasitic agents, spermicides, as well as exhibiting other bioactive functions.

The term "antimicrobial" as used herein means that the polypeptides or proteins of the present invention inhibit, prevent, or destroy the growth or proliferation of microbes such as bacteria, fungi, viruses, or the like.

The term "anti-bacterial" as used herein means that the peptides or proteins employed in the present invention produce effects adverse to the normal biological functions of bacteria, including death or destruction and prevention of the growth or proliferation of the bacteria when contacted with the peptides or proteins.

The term "antibiotic" as used herein means that the peptides or proteins 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 or proteins.

The term "spermicidal" as used herein means that the peptides or proteins employed in the present invention, inhibit, prevent, or destroy the motility of sperm.

The term "anti-fungal" as used herein means that the peptides or proteins employed in the present invention inhibit, prevent, or destroy the growth or proliferation of fungi.

The term "anti-viral" as used herein means that the peptides or proteins employed in the present invention inhibit, prevent, or destroy the growth or proliferation of viruses, or of virally-infected cells.

SUBSTITUTE SHEET (RULE 2B)

-I

~1~ 4a Throughout the description and claims of this specification, the word "comprise" and variations of the word, such as "comprising" and "comprises", means "including but not limited to" and is not intended to exclude other additives, components, integers or steps.

a. a *o S* e S***0 H:\Luisa\Keep\specis\7288.95.MACANI.d 14/05/98 ~C I WO 95/19370 PCTIUS95/00714 The term "anti-tumor" as used herein means that the peptides or proteins inhibits the growth of or destroys tumors, including cancerous tumors.

The term "anti-parasitic" as used herein means that the peptides or proteins employed in the present invention inhibit, prevent, or destroy the growth or proliferation of parasites.

The peptides or proteins 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 or proteins of the present invention allow a method for treating or controlling microbial infection caused by organisms which are sensitive to the peptides or proteins. 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 or proteins.

Because of the antibiotic, antimicrobial, antiviral, and antibacterial properties of the peptides or proteins, they may also be used as preservatives or sterilants or disinfectants of materials susceptible to microbial or viral contamination.

The peptides or proteins 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 or protein 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.

SUBSTITUTE SHEET (RULE 28) 4,- 1.v 0 11 }'I'n PCTI1IQO/flfl71 d The peptides or proteins of the present invention may be administered to a host; in particular a human or non-human animal, in an effective antibiotic and/or anti-tumor and/or anti-fungal and/or anti-viral and/or anti-microbial and/or antibacterial and/or anti-parasitic and/or spermicidal amount.

Depending on the use, a composition in accordance with the invention will contain an effective anti-microbial amount and/or an effective spermicidal amount and/or an effective anti-fungal amount and/or an effective anti-viral amount and/ or an effective anti-tumor amount and/or an effective anti-parasitic and/or an effective antibiotic amount of one or more of the peptides or proteins of the present invention which have such activity. The peptides or proteins may be administered by direct application of the peptides or proteins to the target cell or virus or virally-infected cell, or indirectly applied through systemic administrarlo.n.

The peptides or proteins 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 wound 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, the peptides or proteins increase wound breaking strength- The peptides or proteins of the present invention may also be employed so as to reverse the inhibition of wound healing caused by conditions which depress or compromise the immune system.

The peptides or proteins 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 or proteins may be used to treat skin and burn infections caused by organisms such as, but not limited to, P. aeruqinosa and S. aureus.

SUBSTITUTE SHEET (RULE 28) WO 95/19370 7 PCT/US95/00714 The peptides or proteins 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 aeruqinosa, S aureus, and N. conorrhoeae, by fungi such as but not limited to C. albicans and A fumicatus, by parasites such as but not limited to A. castellani, or by viruses.

The peptides or proteins 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 or proteins 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.

The peptides or proteins may also be employed in treating septic shock in that such peptides neutralize bacterial endotoxins. In general, the peptides or proteins are positively charged, while in general, the bacterial endotoxins are negatively charged. The peptides or proteins are particularly useful in that such compounds neutralize bacterial endotoxins without neutralizing essential proteins in plasma (such as heparin, for example).

The peptides or proteins, when used in topical compositions, are generally present in an amount of at least by weight. In most cases, it is not necessary to employ the peptide in an amount greater than by weight.

In employing such compositions systemically (intramuscular, intravenous, intraperitoneal), the active peptide or protein is present in an amount to achieve a serum level of the peptide of at least about 5 ug/ml. In general, the serum level of peptide or protein need not exceed 500 ug/ ml. A preferred serum level is about 100 ug/ml. Such serum levels may be achieved by incorporating the peptide or protein in a composition to be administered systemically at a dose of from 1 to about 10 mg/kg. In general, the peptide(s) SUBSTITUTE SHEET (RULE 28) WO 95/19370 PCT/US95/00714 8 or protein(s) need not be administered at a dose exceeding 100 mg/kg.

The peptides or proteins 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 or proteins by genetic engineering techniques. The codons encoding specific amino acids are known to those skilled in the art, and therefore DNA encoding the peptides may be constructed by appropriate techniques, and one may clone such DNA into an appropriate expression vehicle a plasmid) which is transfected into an appropriate organism for expression of the peptide or protein.

Upon production or synthesis of the peptide or protein, the N-terminal (NH 2 or amino terminal) of the peptide is reacted such that the lipophilic moiety is attached to the N-terminal of the peptide. For example, the reaction may be a condensation reaction with an amine. When the lipophilic moiety T is 0 O

II

R C the N-terminal is reacted with a carboxylic acid of the formula R-COOH, wherein R is a hydrocarbon having at least 2 carbon atoms. The reaction may be carried out in the presence of a coupling agent, such as, for example, DCC, or DIC, and HOBT. jr in the presence of an acid chloride. Such a reaction results in the formation of an N-terminal substituted peptide or protein having the structural formula hereinabove described.

In one embodiment, x is a peptide which is a basic (positively charged) polypeptide having at least sixteen amino acids wherein the polypeptide includes at least eight hydrophobic amino acids and at least eight hydrophilic amino acids. Still more particularly, the hydrophobic amino acids are in groups of two adjacent amino acids, and each group of SUBSTITUTE SHEET (RULE 28) WO 95/19370 PCT/US95/00714 two hydrophobic amino acids is spaced from another group of two hydrophobic amino acids by at least one amino acid other than a hydrophobic amino acid (preferably at least two amino acids) and generally by no greater than four amino acids, and the amino acids between pairs of hydrophobic amino acids may or may not be hydrophilic.

The hydrophilic amino acids are generally also in groups of two adjacent amino acids in which at least one of the two amino acids is a basic hydrophilic amino acid, with such groups of two hydrophilic amino acids being spaced from each other by at least one amino acid other than a hydrophilic amino acid (preferably at least two amino acids) and generally no greater than four amino acids, and the amino acids between pairs of hydrophilic amino acids may or may not be hydrophobic.

In accordance with a particularly preferred embodiment, the polypeptide comprises a chain of at least four groups of amino acids, with each group consisting of four amino acids.

Two of the four amino acids in each group are hydrophobic amino acids, and two of the four amino acids in each group are hydrophilic, with at least 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 acid.

The hydrophobic amino acids may be selected from the class consisting of Ala, Cys, Phe, Gly, Ile, Leu, Met, Pro, Val, Trp, Tyr, norleucine (Nle), norvaline (Nva), and cyclohexylalanine (Cha). The neutral hydrophilic amino acids may be selected from the class consisting of Asn, Gln, Ser, Thr and homoserine (Hse). The basic hydrophilic amino acids may be selected from the class consisting of Lys, Arg, His, Orn, homoarginine (Har), 2, 4-diaminobutyric acid (Dbu), and p-aminophenylalanine.

Each of the groups of four amino acids may be of the sequence ABCD, BCDA, CDAB, or DABC, wherein A and B are each hydrophobic amino acids and may be the same or different, one of C or D is a basic hydrophilic amino acid, and the other of C or D is a basic or neutral hydrophilic amino acid and may SUBSTITUTE SHEET (RULE 28) c WO 95/19370 10 PCT/US95/00714 be the same or different. In one embodiment, the polypeptide chain may comprise 5 or 6 groups of this sequence. In each group, each of A, B, C and D may be the same in some or all of the groups or may be different in some or all of the groups.

The polypeptide chain preferably has it least 20 amino acids, and no greater than 50 amino acids. It is to be understood, however, that the polypeptide does not have to consist entirely of the groups described above. The polypeptide may have amino acids extending from either or both ends of the noted groups forming the polypeptide chain and/or there may be amino acids between one or more of the at least four groups and still remain within the scope of the invention.

The groups of amino acids may be repeating groups of amino acids, or the amino acids in the v' us groups may vary provided that in each group of the least four groups of amino acids there are two hydrophobic and two hydrophilic amino acids as hereinabove noted.

Thus the biologically active polypeptide may comprise a chain including at least four groups of amino acids, each containing four amino acids. Two of the four amino acids in each group are hydrophobic, at least one amino acid is basic hydrophilic, and the remaining one is basic or neutral hydrophilic, with the polypeptide chain preferably having at least 20 amino acids but no greater than 50 amino acids.

In one embodiment, each of the at least four groups of amino-acids which are in the peptide chain is of the sequence A-B-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 hydrophilic amino acid. The resulting polypeptide chain, therefore, may have one of the following sequences: SUBSTITUTE SHEET (RULE 28) WO 95/19370 PCT/US95/00714 11 a(A-B-C-D)n (Y

(X

2 a(B-C-D-A)n (Y2b

(X

3 )a(C-D-A-B)n(Y 3 )b

(X

4 a(D-A-B-C)n (Y4)b wherein X 1 is D; C-D- or Y 1 is -A or -A-B or

-A-B-C

X

2 is D-A- or C-D-A- Y is -B-C or B-C-D

X

3 is D-A-B- Y3 is -C-D-A

X

4 is A-B-C- Y4 is -D-A-B a is 0 or 1; b is 0 or 1 and n is at least 4.

It is to be understood that the peptide chain may include amino ecids between the hereinabove noted groups of four amino acids provided that the spacing between such groups and the charge on the amino acids does not change the characteristics of the peptide chain which provide amphiphilicity and a positive charge and do not adversely affect the folding characteristics of the chain to that which is significantly different from one in which the hereinabove noted groups of four amino acids are not spaced from each other.

As representative examples of such peptides, there may be mentioned.

I Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser- Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys (SEQ ID NO:1) II Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser- Lys-Als-Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe- Ser-Lys. (SEQ ID NO:2) III Phe-Ser-Lys-Ala-Phe-Ser- Lys-Ala-Phe-Ser-Lys-Ala- Phe-Ser-Lys-Ala- (SEQ ID NO:3) SUBSTITUTE SHEET (RULE 28) i r WO 95/19370 12 PCT/US95/00714 IV -Ser-Lys-Ala-Phe-Ser-Lys-Ala- Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala- Phe-Ser-Lys-Ala-Phe- (SEQ ID NO:4) V Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser- Lys-Ala-Phe-Ser (SEQ ID The peptide may have amino acids extending from either end of the chain. For example, the chains may have a Ser-Lys sequence before the "Ala" end, and/or an Ala-Phe sequence after the "Lys" end. Other amino acid sequences may also be attached to the "Ala" and/or the "Lys" end.

Similarly, in any polypeptide chain having at lease four groups of amino acids of the sequence as described above, the chain may have, for example, a C-D sequence before the first A-B-C-D group. Also other amino acid sequences may be attached to the and/or the end of one of these polypeptide chains. Also there may be amino acids in the chain which space one or more groups of the hereinabove noted four amino acids from each other.

In accordance with another embodiment, X is a magainin peptide.

A magainin peptide is either a magainin such as magainin I, II or III or an analogue or derivative thereof. The magainin peptides preferably include the following basic peptide structure X12 R11-R11-R12-R13-R 11R14-R12-R11 R14-R12-R11-R11-R11-R14a-(R15n-R14a-14wherein R11 is a hydrophobic amino acid, R 12 is a basic hydrophilic amino acid; R 13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid; R 14 and R1 4 a are hydrophobic or basic hydrophilic amino acids; R 15 is glutamic acid or aspartic acid, or a hydrophobic or a basic hydrophilic amino acid, and n is 0 or 1. In a preferred embodiment, R 13 is a hydrophobic or neutral hydrophilic amino acid, R 14 a is a hydrophobic amino acid, and R 15 is glutamic acid or aspartic acid.

SUBSTITUTE SHEET (RULE 28) WO 95/19370 13 PCT/US95/00714 -Thus, for example, a magainin peptide may include the following structure: -Y12-X12 where X12 is the hereinabove described basic peptide structure and Y12 is R2 (ii)

R

1 4a-R 1 2 (iii) RI-RI4a-R12 (iv)

R

14 -R11-R 14 a-R 1 2 where R 11

R

12 R1 4 and R 14 a are as previously defined.

A magainin peptide may also have the following structure: -X12- 12 wherein X12 is as previously defined and Z12 is:

R

16 where R16 is a basic hydrophilic amino acid or asparagine or glutamine.

(ii) R 16

-R

17 where R 17 is a neutral hydrophilic amino acid, a hydrophobic amino acid, or a basic hydrophilic amino acid. Preferably, R17 is a neutral hydrophilic amino acid.

A magainin peptide may also have the following structure: (Y a-X 12-(Z2)b 12 a b where X 12

Y

12 and 212 are as previously defined and a is 0 or 1 and b is 0 or 1.

The magainin peptides may also include the following basic peptide structure X 13 R14-R -R14a

R

2-R 1-R -R2-R13- RII-R 12-R -l-R-R I -R I wherein R 11

R

12

R

13

R

14 and R14 a are amino acids as hereinabove described.

The magainin peptide may also include the following structure X 13

-Z

13 wherein X13 is the hereinabove described basic peptide structure and 213 is

(R

1 1)n-(R 1 1 )n-(R 1 4 an-(R n-(R 1 4 a)n-(R 4 n-

(R

16 17 n wherein R 11

R

14 R1 4 a, R 15

R

16 and R17 are as hereinabove described, and n is 0 or 1, and each n may be the same or different.

SUBSTITUTE SHEET (RULE 218 e -I WO 95/19370 PCT/US95/00714 14 -The magainin peptides generally include at least fourteen amino acids and may include up to forty amino acids.

A magainin peptide preferably has 22 or 23 amino acids.

Accordingly, the hereinabove described basic peptide structures of a magainin peptide may include additional amino acids at the amino end or at the carboxyl end, or at both ends.

As representative examples of such magainin peptides, there may be mentioned peptides having the following primary sequences as given in the accompanying sequence listing as well as appropriate analogues and derivatives thereof: (SEQ ID NO:6) (OH) or (NH 2 (Magainin I) (SEQ ID NO:7) (OH) or (NH 2 (Magainin II) (SEQ ID NO:8) (OH) or (NH 2 (Magainin III) The following are examples of peptide derivatives or analogs of the basic structure: (SEQ ID NO:9) (OH) or (NH 2 (SEQ ID NO:10) (OH) or (NH 2 (SEQ Id NO:11) (OH) or (NH 2 Magainin peptides are described in Proc. Natl. Acad.

Sci. Vol. 84 pp. 5449-53 (Aug. 87). The term "magainin peptides" as used herein refers to the basic magainin structure as well as derivatives and analogs thereof, including but not limited to the representative derivatives or analogs.

In accordance with a further embodiment, X may be a PGLa peptide or an XPF peptide.

A PGLa peptide is either PGLa or an analogue or derivative thereof. The PGLa peptides preferably include the following basic peptide structure X 4: R11-R17-R12 -R11R4-R14-R11

R

11

-R

14

-R

12

-R

11

-R

11

-R

12

-R

11 R11-R11-R12- SUBSTITUTE SHEET (RULE L WO 95/19370 PCT/US95/00714 15 "where P 11

R

12

R

14 and R 17 are as previously defined.

The PGLa peptides generally include at least seventeen amino acids and may include as many as forty amino acids.

Accordingly, the hereinabove described basic peptide structure for a PGLa peptide may include additional amino acids at the amino end or at the carboxyl end or at both the amino and carboxyl end.

Thus, for example, a PGLa peptide may have the following structure: -Y14 14 where X14 is as previously defined and Y14 is

R

1 (ii)

R

1 4

-R

1 1 where R11 and Ri4 are as previously defined.

For example, a PGLa peptide may also have the following structure: -X14-

Z

14where X14 is as previously defined; and Z14 is:

R

11 or (ii) R11-R11 where R 11 is as previously defined.

A PGLa peptide may also have the following structure: (Y4)a-X 4) (Y ~14 14 14 a b where X 14

Y

14 and Z14 are as previously defined, a is 0 or 1 and b is 0 or 1.

An XPF peptide is either XPF or an analogue or derivative thereof. The XPF peptides preferably include the following basic peptide structure X 16

R

11 -R-R 12 -R 1 1

-R-R

14

-R

18

-R

17 RII-R14-R12-R 11 11-R 12 R1-R11-R11-R12-R15)n-

R

1 1 wherein R 1, R 12 R14' R15 and R17 are as previously defined and R18 is glutamine or asparagine or a basic hydrophilic, or hydrophobic amino acid and, n is 0 or 1.

The XPF peptides generally include at least nineteen amino acids and may include up to forty amino acids.

SUBSTITUTE SHEET (RULE 28) WO 95/19370 16 PCT/US95/00714 Accordingly, the hereinabove described basic peptide structure for XPF may include additional amino acids at the amino end, or at the carboxyl end or at both the amino and carboxyl ends.

Thus, for example, an XPF peptide may include the following structure: -Y16-X16 where X16 is as previously defined and Y16 is R11 or (ii) R14-R11 where R11 and R 14 are as previously defined.

An XPF peptide may include the following structure: -X16-Z16where X16 is as previously defined and Z16 is:

R

11 or (ii) R 11

-R

18 or (iii)R 1-R18-Proline; or (iv) R 11

-R

18 -Proline-R 12 An XPF peptide may also have the following structure:

(Y

16 )a-X16-(Z16)b where X 16

Y

16 and Z16 are as previously defined: a is 0 or 1 and b is 0 or 1.

Preferred are XPF or PGLa peptides, which are characterized by the following primary amino acid sequences as given in the accompanying sequence listing: PGLa: (SEQ ID NO:12) (NH 2 XPF: (SEQ ID NO:13) 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.

In accordance with yet another embodiment, X is a CPF peptide or appropriate analogue or derivative thereof.

CPF peptides as well an analogues and derivatives thereof are herein sometimes referred to collectively as CPF peptides.

SUBSTITUTE SHEET (RULE 28) i Wd 95/19370 17 I'CT/US9S/00714 -The CPF peptide may be one which includes the following basic peptide structure -R21-R21-R22-R22- R 2221-R21-R23-R21 -R 21-R1-R-R -R-R 1-R -R -R wherein R 21 is a hydrophobic acid; R 22 is a hydrophobic amino acid or a basic hydrophilic amino acid; R23 is a basic hydrophilic amino acid; R24 is a hydrophobic or neutral hydrophilic amino acid; and

R

25 is a basic or neutral hydrophilic amino acid.

The hereinabove basic structure is hereinafter symbolically indicated as X 20 The hydrophobic amino acids are Ala, Cys, Phe, Gly, Ile, Leu, Met, Val, Trp, Tyr, norleucine (Nle), norvaline (Nva), and cyclohexylalanine (Cha).

The neutral hydrophilic amino acids are Asn, Gin, Ser, Thr, and homoserine (Hse).

The basic hydrophilic aminc ac3ds are Lys, Arg, His, Orn, honoarginine (Har), 2,4-di dinibutyric acid (Dbu), and p-aminophenylalanine.

The CPF peptide may include only the hereinabove noted amino acids or may include additional amino acids at the amino and/or carboxyl end or both the amino and carboxyl end.

In general, the peptide does not include more than 40 amino acids.

The CPF peptides including the above basic structure preferably have from 1 to 4 additional amino acids at the amino-end.

Accordingly, such preferred peptides may be represented by the structural formula: wherein X20 is the hereinabove described basic peptide structure and Y20 is

R

2 5 or (ii) R 2 2

-R

2 5 or SUBSTITUTE SHEET (RULE 28) WVi O5/19371 I)i/t f tne mnI11 A rJv O5II _A t18 UUis AI YJ UUI A -(iii)R21-R22-R25; or (iv) R 2 2

-R

2 1

-R

2 2 -R25; preferably Glycine R21-R22-R25.

wherein R21' R22 and R25 are as previously defined.

The carboxyl end of the basic peptide structure may also have additional amino acids which may range from 1 to 13 additional amino acids.

In a preferred embodiment, the basic structure may have 1 to 7 additional amino acids at the carboxyl end, which may be represented as follows: 20 wherein X is the hereinabove defined basic peptide structure and is

R

2 1 or (ii) R 2 1

-R

2 1 or (iii)R 2 1

-R

2 1

-R

2 4 or (iv) R 2 1

-R

2 1

-R

2 4

-R

2 4 or

R

2 1

-R

2 1

-R

2 4

-R

2 4

-R

2 6 or (vi) R 21

-R

21

-R

24

-R

24

-R

26 -Gln; or (vii)R21-R21-R24-R24-R26-Gln-Gln, wherein R21 and R24 are as previously defined, and R 26 is proline or a hydrophobic amino acid.

Preferred peptides may be represented by the following structural formula 20 (Z 2 0 )b wherein X 2 0

Y

2 0 and Z20 are as previously defined and a is 0 or 1 and b is 0 or 1.

.Representative examples of CPF peptides which may be employed, some of which have been described in the literature, include the following sequences as given in the accompanying sequence listing: (SEQ ID NO:14) (SEQ ID (SEQ ID NO:16) (SEQ ID NO:17) (SEQ ID NO:18) (SEQ ID NO:19) SUBSTITUTE SHEET (RULE 28) WO95/9370 PCTISq/0nn714 -19- (SEQ -ID (SEQ ID NO:21) (SEQ ID NO:22) (SEQ ID NO:23) (SEQ ID NO:24) (SEQ ID (SEQ ID NO:26) A review of the CPF peptides can be found in Richter, K., Egger, and Kreil (1986) J. Biol. Chem. 261, 3676-3680; Wakabayashi, Kato, and Tachibaba, S. (1985) Nucleic Acids Research 13, 1817-1828; Gibson, Poulter, L., Williams, and Maggio, J.E. (1986) J. Biol. Chem. 261, 5341-5349.

In accordance with yet another embodiment, X is a peptide which includes one of the following basic structures

X

3 1 through X 3 7 wherein: X31 is [R 3 1 -R32-R32-R33-R31-R32-R32 n X32 is -[R32-R32-R33-R31-R32-R32-R31 n' X33 is [R 3 2

-R

3 3

-R

3 1 R3- R 3 2-R 3 1

-R

3 2 -n; X34 is [R 33

-R

31

-R

3 3 2 2-R 31

-R

32

-R

32 -n is [R 3 1

-R

3 2

-R

3 2

R

3 1

-R

3 2

-R

3 2

-R

3 3 1-n; X36 is -[R 3 2

-R

3 2

-R

3 1

-R

3 2 3 -R32R 3 3

-R

3 1 and X37 is -[R 3 2 -R 31-R32-R32-R33-R31-R32 n wherein R 3 1 is a basic hydrophilic amino acid, R 32 is a hydrophobic amino acid, R 33 is a neutral hydrophilic, basic hydrophilic, or hydrophobic amino acid, and n is from 1 to The basic hydrophilic amino acids may be selected from the class consisting of Lys, Arg, His, Orn, homoarginine (Har), 2,4-diaminobutyric acid (Dbu), and paminophenylalanine.

The hydrophobic amino acids may be selected from the class consisting of Ala, Cys, Phe, Gly, Ile, Leu, Met, Pro, Val, Trp and Try, norleucine (Nle), norvaline (Nva), and cyclohexylalanine (Cha).

The neutral hydrophilic amino acids may be selected from the class consisting of Asn, Gin, Ser, Thr, and homoserine (Hse).

SUBSTITUTE SHEET (RULE 28) W ri or5/1937n PCT/US95/I0714 20 -In accordance with one embodiment, when the peptide includes the structure X the peptide may include the following structure:

Y

31

-X

31 wherein X31 is as hereinabove described, and Y31 is: R32; (ii) R32-R32; (iii) R31-R32-R32; (iv) R 3 3

-R

3 1

-R

3 2

-R

3 2

R

3 2

-R

3 3

-R

3 1

-R

3 2

-R

3 2 or (vi) R 32

-R

32

-R

33

-R

31

-R

32

-R

32 wherein R 31

R

32 and R 33 are as hereinabove described.

In accordance with another embodiment, when the peptide includes the structure X 31 the peptide may include the following structure: X31-Z31, wherein X31 is as hereinabove described, and Z31 is:

R

3 1 (ii) R31-R32; (iii)R 3 1

R-R

3 2 (iv) R 3 1

-R

3 2

-R

3 2

-R

3 3

R

3 1

-R

3 2

-R

3 2

-R

3 3

-R

3 1 or (vi) R 3 1

-R

3 2

-R

3 2 3 3 -R3 3 1

-R

3 2 In accordance with yet another embodiment, the peptide may include the following structure: (Y31)a-X31- (Z31) b wherein Y31 and Z31 are as previously defined, a is 0 or 1, and b is 0 or 1.

.When the peptide includes the structure X 32 the peptide may include the following structure: Y32 X 32 wherein X32 is as hereinabove described, and Y32 is: R31; (ii) R32-R31; (iii)R3 2

-R

32

-R

31 (iv) R 3 1

-R

3 2

-R

3 2

-R

3 1 SUBSTITUTE SHEET (RULE 26) St/A OC/1071 't PCT/Ir)S5/n714 21

R

3 3

-R

3 1

-R

3 2

-R

3 2

-R

3 1 or (vi) R32-R33-R3 -R32-R3-R1 In another embodiment, when the peptide includes the structure X32' the peptide may include the following structure: X32 Z 32 wherein X32 is as hereinabove described, and Z32 is: R32; (ii) R32-R32; (iii)R32-R32-R33; (iv) R 32

-R

32

-R

33

-R

31

R

3 2

-R

3 2

-R

3 3

-R

3 1

-R

3 2 or (vi) R 3 2

-R

3 2

-R

3 3

-R

3 1

-R

3 2

-R

3 2 In accordance with yet another embodiment, the peptide may include the following structure: (Y32a X32 (Z 3 2 wherein Y32 and Z32 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 33 the peptide may include the following structure: Y33 X33 wherein X33 is as hereinabove described, and Y33 is: R32; (ii) R31-R32; (iii)R 3 2

-R

3 1 -R32; (iv) R32-R32-R31-R32;

R

31

-R

32

-R

32

-R

31

-R

32 or .4vi) R 3 3

-R

3 1

-R

32 -2-R3-R 3 1

-R

3 2 wherein R 3 1

R

3 2 and R33 are as hereinabove described.

In accordance with another embodiment, when the peptide includes the structure X 33 the peptide may include the following structure: X33 33 wherein X33 is as hereinabove described, and Z33 is: R32; (ii) R 3 2

-R

3 3 SUBSTITUTE SHEET (RULE 26) I I W A Inie/ PCT/ITS)Q /(i7 4 d v v L/ j 1 JiU 2 2 (iii)R32-R33-R31 (iv) R32-R33-R31-R32 R32-R33-R31-R32-R32 or (vi) R32-R33-R31-R32-R32-R31' In accordance with yet another embodiment, the peptide may include the following structure: (Y33)a X33 (Z33)b, wherein Y33 and Z33 are as previously defined, a is 0 or 1, and b is 0 or 1.

In accordance with yet another rs-bodiment, when the peptides includes the structure X 34 the peptide may include the following structure: Y34 X 3 4 wherein X34 is as hereinabove described, and Y34 is:

R

3 2 (ii) R32-R32; (iii)R31-R32-R32 (iv) R 3 2

-R

3 1

-R

3 2

-R

3 2

R

3 2

-R

3 2

-R

3 1

-R

3 2

-R

3 2 or (vi) R 3 1

-R

3 2

-R

3 2

-R

3 1

-R

3 2

-R

3 2 wherein R 3 1' R 3 2 and R33 are as hereinabove described.

In accordance with another embodiment, when the peptide includes the structure X 3 4 the peptide may include the following structure: X34-Z34, wherein X3 is as hereinabove described, and Z34 is: R33; (ii)

R

3 3

-R

3 1 4iii)R33-R31-R32; (iv) R 3 3

-R

3 1

-R

3 2

-R

32

R

3 3

-R

3 1

-R

3 2

R

3 2

-R

3 1 or (vi) R 3 3

-R

3 1 -32-R 3 2

-R

3 1

-R

3 2 In accordance with yet another embodiment, the peptide may include the following structure:

(Y

34 )a X34 (Z34)b, wherein X34 and Z34 are as previously defined, a is 0 or 1, and b is 0 or 1.

SUBSTITUTE SHEET (RULE 26) WO 95/19370 PCT/US95/00714 23 In accordance with a further embodiment, when the peptide includes the structure X 35 the peptide may include the following structure: Y35-X35, wherein X35 is as hereinabove described, and is: R33; (ii) R 32

-R

33 ('.ii)R 3 2

-R

3 2

-R

3 3 (iv) R 3 1

-R

3 2

-R

3 2

-R

3 3; R 32-R 31-R 32-R 32 -R 33 or (vi) R 32

-R

32

-R

31

-R

32

-R

32

-R

33 wherein R31, R32 and R33 are as hereinabove described.

In accordance with another embodiment, when the peptide includes the structure X 35 the peptide may include the following structure:

Z

3 5 wherein X35 is as hereinabove described, and is: R31 (ii) R31-R32; (iii)R 31

-R

32

-R

32 (iv) R 31

-R

32

-R

32

-R

31

R

3 1

-R

3 2-R 3 2

-R

3 1

-R

3 2 or (vi) R 31

-R

32

-R

32

-R

31

-R

32

-R

32 In accordance with yet another embodiment, the peptide may include the following structure:

(Y

3 5 )a X 3 5 (Z35)b' wherein X35 and Z35 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 X36, the peptide may include the following structure: Y36 X36' wherein X36 is as hereinabove described, and Y36 is: R31; (ii)

R

3 3

-R

3 1 (iii)R 3 2 3 3 -R3 3 1 (iv) R 32

-R

32

-R

33

-R

31 SUBSTITUTE SHEET (RULE 26) k Wd 95/19370 24 PCT/US95/00714 R31-R32-R32-R 33

-R

31 or (vi) R 3 2 -R31-R 3 2

-R

3 2 -R33-R 3 1 wherein R 3 1

R

3 2 and R 33 are as hereinabove described.

In accordance with another embodiment, when the peptide includes the structure 36, the peptide may include the following structure: X36-Z36, wherein X36 is as hereinabove described, and Z36 is: R32; (ii) R32-R32; (iii)R 3 2

-R

3 2

-R

3 1 (iv) R32-R32-R31-R32;

R

3 2

-R

3 2

-R

3 1

-R

3 2

-R

3 2 or (vi) R 32 32

-R

31

-R

32

-R-RR

33 In accordance with yet another embodiment, the peptide may include the following structure:

(Y

3 6 )a X 3 6 (Z36)b, wherein X36 and Z36 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 X37, the peptide may includes the structure Y 37

-X

37 wherein X37 is as hereinabove described, and Y37 is:

R

3 2 (ii) R31-R32; (iii)R 33-R31-R32; (iv) R 3 2

-R

3 3

-R

3 1

-R

3 2

R

3 2

-R

3 2

-R

3 3

-R

3 1

-R

3 2 or !vi) R 3 1

-R

3 2

-R

3 2

-R

3 3

-R

3 1

-R

3 2 wherein R 3 1

R

3 2 and R33 are as hereinabove described.

In accordance with another embodiment, when the peptide includes the structure X 37 the peptide may include the following structure: X37 Z 3 7 wherein X37 is as hereinabove described, and Z37 is: R32; (ii) R 3 2

-R

3 1 SUBSTITUTE SHEET (RULE 28) IWO1 95/19370 PTU9/014-2 25 PCTIUS95/00714 (iii)R 3- R31R32 (iv) R 3 R 3 -R 3 R 32 M R32- 31- 32- 3 2

R

33 or (vi) R 32

-R

31

R

3 2

R

3 2 R 3- R 1 In accordance with yet another embodiment, the peptide may include the following structure: (Y 37 )a X 37 Z 3)b' wherein X 7and Z 7are as previously defined, a is 0 or 1, and b is 0 or 1.

In a preferred embodiment, n is 3, and most preferably the peptide is of one of the following structures as given in the accompanying sequence (Lys Ile Ala (Lys Ile Ala (Lys Ile Ala (Lys Leu Ala (Lys Phe Ala (Lys Ala Leu (Lys Leu Leu (Lys Ala Ile (Gly Ile Ala (Lys Ile Ala (Gly Ile Ala (Lys Phe Ala (Giy Phe Ala (Lys Ile Ala (Lys Ile Ala (Orn le Ala lGly Ile Ala (Lye Nie Ala (Lys Nie Ala (Lys Ile Ala (Lys Nva Ala (Lys Nva Ala (Lys Leu Leu (Lys Leu Leu (Lys Ile Ala Gly Lys Lys Ile Gly Lys Gly Lye Gly Lys Ser Lys Lys Ala Gly Lys Lys Ile Lys Ile Arg Ile Arg Ile Lys Ile Gly Orn Arg Ile Gly Lys Arg Ile Gly Lys Gly Lye Gly Lys Gly Lys Gly Lys Ser Lye Ser Lye Gly Lys listing: Ile Ala) 3 Ala Gly) 3 Ile Giy) 3 Leu Ala) 3 Phe Ala) 3 Ala Leu) 3 Leu Giy) 3 Ala Ilie) 3 Ala Lys) 3 Phe Gly) 3 Ala Lye) 3 Ala Gly) 3 Ala Lye) 3 Ile Ala) 3 Ala Gly) 3 Ile Ala)3 Phe Lye) 3 Nie Ala) 3 Ile Ala)3 Nie Ala) 3 Nva Ala) 3 Ile Ala) 3 Leu Gly) 3 Phe Gly) 3 Nva Ala) 3

(SEQ

ID NO: 27) ID NO: 28) ID NO: 29) ID NO: ID NO: 31) ID NO: 32) ID NO: 33) ID NO: 34) ID NO: ID NO: 36) ID NO: 37) ID NO: 38) ID NO: 39) ID NO: ID NO: 41) ID NO: 42) ID NO: 43) ID NO: 44) ID NO: ID NO: 46) ID NO: 47) ID NO: 48) ID NO: 49) ID NO: ID NO: 51) SUBSTITUTE SHEET (RULE Wci 95/19370 PTU9IOI PCTIUS95/00714 26 (His Ile Ala Gly His Ile Ala) 3 (Ala GJly Lys (Ile Ala Lys (Lys Tle Ala (Arg Ile ALa (Lys Val Ala (Lys Ile Ala (Ala Lys Ile (Orn Ile Ala (Lys Phe Ala (Lys Ile Ala (Lys Cha Ala (Lys Nie Ala (Arg Ile Ala (Har Ile Ala (Xaa Ile Ala (Lys Ile Ala In (SEQ ID NO:G7) Ile Ile Gly Gly Gly Gly Ala Gly Gly Gly Gly Lys Gly Giy Giy Gly Ala Lys Ala Gly Arg Ile Arg Ile Lys Ile Lys Val Gly Lys Orn Ile Lys Ile Lys Phe Lys Ile Ile Ala Lys Ile Har Ile Lys Ile Xaa Ile Ile)3 Lys) 3 Ala)3 Ala)3 Ala)3 Ala)3 Ile)3 Ala)3 Ala)3 Ala)3 Ala)3 Gly) 3 Ala)3 Al.a)3 Ala)3 Ala)3

(SEQ

ID NO: 52) ID NO: 53) ID NO: 54) ID NO: ID NO: 56) ID NO: 57) ID NO: 58).

ID NO: 59) ID NO: ID NO: 61) ID NO: 62) ID NO: 63) ID NO: 64) ID NO: ID NO: 66) ID NO: 67) ID NO: 68) and (SEQ ID NO:68), Xaa is p-aminophenyialanine.

In accordance with another embodiment, X is a peptide which includes the following basic structure X4 R31- 32- 32- 33- 34- 32 R32- 31- 32- 32- 32- 34- 32- 32' wherein R 31 R 32 and R 33 are as hereinabove described, and R 4is a basic hydrophilic or hydrophobic amino acid.

In accordance with one embodiment, the peptide may include the following structure: Y 4-x 0'wherein X 0is as hereinabove described, and Y (ii) R3- 2 (iii)R 34

R

32

R

32 (iv)R33R34R32R32

R

32

-R

32

-R

34

-R

32

-R

32 SUBSTITUTE SHEET (RULE WiOr n13 n7ii n "nb^lrnnlT nnn Ti A Wv v ,JI~ JJ,.J I 27 -I (vi) R 32

-R

3 2-R 3

R

34

-R

32

-R

32 or (vii)R31-R32-R32-R33R-R34-R32'-R32 wherein R31' R 32 and R33 and R34 are as hereinabove described.

In accordance with another embodiment, X is a peptide which includes the following structure: X40-Z40, wherein X40 is as hereinabove described, and is: R31; (ii) R3-R32 (iii)R 31

-R

32

-R

32 (iv) R31-R32-R32-R33' R31-R32-R32-R33-R34' (vi) R 3 1

-R

3 2

-R

3 2

-R

3 3

-R

3 4

-R

3 2 or (vii)R 3-R-R32-R33-R34-R32-R32, wherein R 31

R

32 and R33' and R34 are as hereinabove.described.

In accordance with yet another embodiment the peptide may include the following structure: (Y40)a-X40-(Z40)b' wherein Y40 and Z40 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 given in the accompanying sequence listing: (SEQ ID NO:69) In another preferred embodiment, the peptide has the following structural formula as given in the accompanying sequence listing: (SEQ ID In accordance with a further embodiment, the peptide has one of the one of the following structural formulae as given in the accompanying sequence listing: (SEQ ID NO:71) (SEQ ID NO:72) (SEQ ID NO:73) (SEQ ID NO:74) (SEQ ID (SEQ ID NO:76) (SEQ ID NO:77) SUBSTITUTE SHEET (RULE 28) WO 95/19370 28 PCT/US95/00714 (SEQ ID NO:78) (SEQ ID NO:79) (SEQ ID (SEQ ID NO:81) (SEQ ID NO:82) (SEQ ID NO:83) (SEQ ID NO:84) (SEQ ID In accordance with another embodiment, X is a peptide which includes one of the following structural formulae: (Lys Ile Ala Lys Lys Ile Ala)-n, (ii) (Lys Phe Ala Lys Lys Phe Ala, and (iii) (Lys Phe Ala Lys Lys Ile Ala) wherein n is from 1 to Preferably, n is 3, and the peptide has one of the following structural formulae: (Lys Ile Ala Lys Lys Ile Ala) 3 (SEQ ID NO:86) (Lys Phe Ala Lys Lys Phe Ala) 3 (SEQ ID NO:87) (Lys Phe Ala Lys Lys Ile Ala) 3 (SEQ ID NO:88) In accordance with another embodiment, the X is a peptide which is selected from the group consisting of the following structural formulae as given in the accompanying sequence listing: (SEQ ID NO:89) (SEQ ID (SEQ ID NO:91) (SEQ ID NO:92) In accordance with yet another embodiment, X is a cecropin or sarcotoxin.

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.

SUBSTITUTE SHEET (RULE 28) Wn 95/19370 PCT/US9S/00714 29 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.

In accordance with another embodiment, X is melittin or an analogue or derivative thereof.

Melittin is an amphipathic peptide consisting of 26 amino acid residues, a.,d is isolated from honeybee (Apis mellifera) venom. Habermann, et al., HopDe-Seyler's Zeitschrift Phvsiol. Chem., Vol. 348, pgs 37-50 (1987).

Melittin has the following structural formula as represented by the three-letter amino acid code: Gly Ile Gly Ala Val Leu Lys Val.Leu Thr Thr Gly Leu Pro Ala Leu Ile Ser Trp Ile Lys Arg Lys Arg Gln Gln (SEQ ID NO:93) In another embodiment, X is a amphiphilic peptide which includes the following basic structure X 50 R41-R42-R42-R41-R42-R42-R41-R41-R42-R41-R 41' R41 is a hydrophobic amino acid, and R 42 is a basic hydrophilic or neutral hydrophilic amino acid.

In one embodiment, the peptide includes the basic structure Y 50

-X

5 0 wherein X50 is as hereinabove described and is: R41 (ii) R 42

-R

41 or (iii)R 2-R 42 -R 4, wherein R41 and R42 are as hereinabove described.

In one embodiment, R 41 is leucine. In another embodiment, R 42 is lysine. Representative examples of peptides in accordance with this aspect of the present invention include those having the following structures: SUBSTITUTE SHEET (RULE 26) W WO 95/1937n PCT/US /5/nfn71 30 (SEQ ID NO:94) (SEQ ID (SEQ ID NO:96) (SEQ ID NO:97) In accordance with another embodiment, X is an amphiphilic peptide which includes the following basic structure X52: R42-R41-R42-42-R41-R41-R42-R42-R41-R42- R 4 2 wherein R 41 is a hydrophobic amino acid and R 42 is a basic hydrophilic or neutral hydrophilic amino acid.

In one embodiment R 41 is leucine. In another embodiment, R 42 is lysine.

In one embodiment, the peptide includes the basic structure Y 52

-X

5 2 where X52 is as hereinabove described, and Y52 is: R42 (ii) R41-R42; (iii) R41-R41-R42; (iv) R 4 2

-R

4 1

-R

4 1

-R

4 2 or R42-R42-R41-R41-R42' In one embodiment, the peptide may have the following structure; Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu Arg Arg (SEQ ID NO:98) .In another embodiment, the peptide includes the basic structure X 52

-Z

52 where X52 is as hereinabove described, and Z52 is: R41 (ii) R41-R41; (iii) R41-R41-R42' (iv) R 4 1

-R

4 1

-R

4 2

-R

4 2 or R41-R41-R42-R42-R41' In one embodiment, the peptide may have the following structure: SUBSTITUTE SHEET (RULE 28) I W695/19370 31 PCT/US95/00714 Lys Leu Lys L/s Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu 10 (SEQ ID NO:99) In another embodiment, the peptide may include the structure: (Y52)a X52 (Z 5 2 wherein X 52 Y52 and Z52 are as hereinabove described, and a is 0 or 1, and b is 0 or 1.

In another embodiment X is a biologically active amphiphilic peptide which includes the following basic structure X 54 R41-R42-R R41 41-R42-R42-41-42-R42-R41-R41-R42-R4243' wherein R 41 and R 42 are as hereinabove described, and R 43 is a neutral hydrophilic amino acid.

In one embodiment, the peptide may have the following structure: (SEQ ID NO:100) In another embodiment, the peptide may have the following structure: (SEQ ID NO:101) In another embodiment, X is a biologically active amphiphilic peptide which includes the following basic structure X 56 R41-R42-R41-R41-R42-R42-R41-R41-R42-R42

R

44 wherein R41 and

R

42 are as hereinabove described, and R 44 is a neutral hydrophilic amino acid or proline.

In one embodiment, the peptide may include the structure: X56-Z56, wherein X56 is as hereinabove described, and Z56 is: -R42; (ii) -R42-R42; (iii) -R 4 2

-R

4 2

-R

4 1 (iv) -R 4 2

-R

4 2

-R

4 1

-R

4 1 -R42-R42-R4-R1-1-R42 (vi) -R 42

-R

4 2

-R

4 4 1

R-R

42

-R

42 or (vii) R 42

-R

42

-R

41

-R

41

-R

42

-R

42

-R

4 1 SUBSTITUTE SHEET (RULE 26) SWO 95/19370 PrT/iSo95/n0714 32 In a preferred embodiment, the peptide may have one of the following structures: (SEQ ID NO:102); or (SEQ ID NO:103).

In another embodiment, X is a biologically active amphiphilic peptide which includes the following basic structure X58: R41-R41-R42-R 42-R 41-R 42-R 42-R 41-R41-R42-R42-R 4 1

-R

4 3 wherein

R

4 1

R

4 2 and R 4 3 are as hereinabove described.

In one embodiment, the peptide includes the structure X58-Z58, wherein X58 is as hereinabove described, and Z58 is: -R41; (ii) -R41-R45; (iii) -R41-R45-R45' (iv)

-R

4 1

-R

4 5

-R

4 5

-R

4 3

-R

4 1

-R

4 5

-R

4 5

-R

4 3

-R

4 1 (vi)

-R

4 1 -R45-R 4 5

-R

4 3

-R

4 1

-R

4 3 (vii)

-R

4 1

-R

4 5

-R

4 5

-R

4 3

-R

4 1

-R

4 3

-R

4 3 (viii) -R 4 1

-R

4 5

-R

4 5

-R

4 3

-R

4 1

-R

4 3

-R

4 3

-R

4 5 or (ix) -R 4 1

-R

4 5

-R

4 5

-R

4 3

-R

4 1

-R

4 3

-R

4 3

-R

4 5

-R

4 3 wherein R41 and R43 are as hereinabove described, and R 4 5 is proline.

In one embodiment, the peptide.has the following structure: (SEQ ID NO:104).

In another embodiment, X is a biologically active amphiphilic peptide which includes the following basic structure R 41-R 41-R -R 42-R 4-R 41-R 4-R 41-R 41-R 4-R 42-R 41-R 4-R 42-R 41 41 43 42 41 41 41. 41 41 41 42 41 41 42 42 R41-R41-R42-R42-R41-, wherein R41, R 42 and R3 are as hereinabove described. In one embodiment, the peptide may have the following structure: (SEQ ID NO:105).

In accordance with another embodiment, X is a peptide which includes the following basic structure X62 -R41-R42-R 42-R 41-R 42-R 42-R41 wherein R 4 1 and R 42 are as hereinabove described.

SUBSTITUTE SHEET (RULE 26) WO'519Q'370in ~~mnrnnrlrrr r 33 ri iu 1A/1q In one embodiment the peptide includes the following structure Y62 X 62 where X62 is as hereinabove described, and Y6 is: 62 R41 (ii)

R

4 2-R 4 1 (iii) R42-R42-R41; or (iv)

R

41

-R

42

-R

42 -R41' Representative examples of such peptides include the following, the sequences of which are given in the accompanying sequence listing: (SEQ ID NO:106) (SEQ ID NO:107) (SEQ ID NO:108) (SEQ ID NO:109) (SEQ ID NO:110) (SEQ ID NO:111) In one embodiment, the peptide includes the structure X62-Z62, wherein X62 is as hereinabove described, and Z62 is: R4 (ii) R41-R42 (iii) R41-R42-R42; or (iv) R 41

-R

42

-R

42

-R

41 where R 4 1 and R 42 are as hereinabove described.

A representative example includes the following peptide having the structural formula given below and listed in the accompanying sequence listing: (SEQ ID NO:112) In another embodiment, the peptide has the structure (Y62)a X62- (Z62)b' wherein X62 Y62 and Z62 are as hereinabove described, a is 0 or 1, and b is 0 or 1.

Representative examples of such peptides include the following, the structures of which are given in the accompanying sequence listing: (SEQ ID NO:113) (SEQ ID NO:114) (SEQ ID NO:115) (SEQ ID NO:116) SUBSTITUTE SHEET (RULE 28) WOi 95/19370 PDrTTIOS95/0071 34 In another embodiment X is a peptide having the following structural formula: (SEQ ID NO:117) In another embodiment, X is a biologically active amphiphilic peptide including the following basic structure X64: -R42-R42-R42-R41-R41-R42-R42-R41-' wherein R 4 1 and R 42 are as hereinabove described.

In one embodiment, the peptide may include the structure Y64-X64-, wherein X64 is as hereinabove described, and Y64 is:

-R

4 1 or (ii)

R

4 2-R 4 1 In another embodiment, the peptide may include the structure X 64

-Z

64 wherein X64 is as hereinabove described, and Z64 is: R42 (ii) R 42

-R

42 or (iii) R42-R42-R41i In yet another embodiment, the peptide has the structure: (Y64)a-X64-(Z64)b' wherein X 64 Y64' and Z64 are as hereinabove described, a is 0 or 1, and b is 0 or 1.

Representative examples of such peptides include the following: (SEQ ID NO:127) (SEQ ID NO:128) (SEQ ID NO:129) In yet another embodiment, X is a biologically active amphiphilic peptide including the following basic structure X66: R41-R42-R42-R41-R41-R46-R42-R41-42-42-R41 wherein R41 and R 42 are hereinabove described and R 46 is glutamic acid.

A representative example of such a peptide is the following: (SEQ ID NO:130) SUBSTITUTE SHEET (RULE 28) Wn'9/I9370 PrT/nI.S q/0f071 35 In yet another embodiment, X is a biologically active amphiphilic peptide including the following basic structure X68: -R42-R42-R41-R41-R42-R46-R41-R42-R42-R41-' wherein R41' R42, and R 46 are hereinabove described.

In one embodiment, the peptide includes the following basic structure Y68-X68, wherein X68 is as hereinabove described, and Y68 is:

R

41 Representative examples of such peptides include the following: (SEQ ID NO:131) (SEQ ID NO:132).

In another embodiment, X is a biologically active amphiphilic peptide including the following basic structure

X

-R41-R42-R42-R41-R41-R42-R -R 41-R 42-R 41-R41-' wherein R 41 and R 42 are hereinabove described. A representative example of such a peptide has the following structure: (SEQ ID NO:133).

In another embodiment, X is a biologically active amphiphilic peptide including the following basic structure X72: -R42-R42-R41-R41-R42-R47-R41-R42-R42-R41-, wherein R41 and R 42 are hereinabove described, and R 4 7 is aspartic acid.

A representative example of such a peptide has the following structure: (SEQ ID NO:134).

In yet another embodiment, X is a biologically active amphiphilic peptide having the following structure: (SEQ ID NO:135).

In yet another embodiment, X is a biologically active amphiphilic peptide including the following structure X 74 SUBSTITUTE SHEET (RULE 26) w1 r1/111n A PTIr/I coQ/l'71 1 A

I

3 6 R42-R41-R42-R41-41-R42-R42-R41-R46-R42-R 4 1 wherein R41' R 4 2 and R46 are hereinabove described. A representative example of such a peptide has the following structure: (SEQ ID NO:136).

In another embodiment, X is a biologically active amphiphilic peptide including the following structure X 76 -R41-R42-R421-R 4 1 -R41-R 4 2 wherein R41 and R42 are hereinabove described.

In another embodiment, the peptide includes the structure Y 76

-X

76 wherein X76 is as hereinabove described, and Y76 is: -R42 (ii)

-R

4 2-R 4 2; (iii) -R41-R42-R42; (iv) -R41-R41-R42-R42;

-R

4 2

-R

4 1

-R

4 1

-R

4 2

-R

4 2 or (vi) -R42-R42-R41-R41-R42-R42 In another embodiment, the peptide includes the structure -X 7 6

-Z

7 6 wherein X76 is as hereinabove described, and Z76 is: R48- (ii) R 4 8

-R

4 1 or (iii) R 4 8

-R

4 1

-R

4 2 wherein R 4 1 and R 4 2 are as hereinabove described, and R 4 8 is a basic hydrophilic, neutral hydrophilic, or hydrophobic amino acid.

In yet another embodiment, the peptide has the following structural formula: (Y76)a-X76-(Z76)b, wherein X 7 6, Y 76 and Z76 are as hereinabove described, a is 0 or 1, and b is 0 or 1.

Representative examples of such peptides include the following: (SEQ ID NO:137) (SEQ ID NO:138) (SEQ ID NO:139).

SUBSTITUTE SHEET (PFULE 28) WO 9e/130211 Yi/'tmrirtf mnncl V- I7.'U 37 DIU VIJU/ I 4 In yet another embodiment, X is a biologically active amphiphilic peptide including the following structural formula X 78 -R41-R42-R41-R41-R 42

-R

42 -R41-R2-R42-R1' wherein R41 and R42 are as hereinabove described. A representative example of such a peptide has the following structure: (SEQ ID NO:140).

In another embodiment, X has the following structure: (SEQ ID NO:149).

In another embodiment, X is a biologically active amphiphilic peptide including the following structural formula X 80 -R41-R42-R42-41-R41-R42-R46-R41-R41-R42-R41-, wherein R41' R 42 and R46 are as hereinabove described. A representative example of such a peptide has the following structure: (SEQ ID NO:151) In accordance with yet another embodiment, X is an ion channel-forming peptide or protein.

Ion channel-forming proteins or peptides which may be employed 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. _xP. 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 if the ion channel-forming proteins as well as analogues and derivatives.

SUBSTITUTE SHEET (RULE 28) I I W 95/l13'7n0 rrMn cn* 38 ;IU F-J It In accordance with yet another embodiment, each of the amino acid residues of the peptides or proteins may be a Damino acid 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 or proteins, when consisting entirely of D-amino acid or glycine residues, may have increased resistance to proteolytic enzymes while retaining their activity. Such peptides thus may be administered orally. Also, in accordance with another embodiment, all of the amino acid residues may be D-amino acid or glycine residues, or L-amino acid or glycine residues.

It is also to be understood that the peptides or proteins may be administered in combination with one another.

In accordance with another embodiment, the N-terminal substituted peptides or proteins of the present invention may be employed in combination with an ion having phamacological properties for the purposes hereinabove described.

An ion having pharmacological properties is one which when introduced into a target cell or virus or virally-infected cell inhibits and/or prevent and/or destroys the growth of the target cell, virus or virally-infected cell.

Such an ion having pharmacological properties is cne which in the absence of an ion channel forming peptide is unable to cross a natural or synthetic lipid membrane; in particular a cell or virus membrane, in sufficient amounts to affect a cell or virus adversely.

The peptide or protein 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 or protein and ion having pharmacological properties. As representative examples of ions having pharmacological properties which may be employed, there iay be mentioned fluoride, peroxide, bicarbonate, silver, zinc, mercury, arsenic, copper, SUBSTITUTE SHEET (RULE 28) WI 95/19370 PCT/US9S/nn714 39 platinum, antimony, gold, thallium, nickel, selenium, bismuth, and cadmium ions.

The peptide or protein and the ion having pharmacological properties, whether administered or prepare".

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, the amount of ion is effective to reduce the maximum effective concentration of the peptide or protein for inhibiting growth of a target cell, virus, or virally-infected cell.

The ion having pharmacological properties, when used topically, is generally employed in a concentration of from 0.05% to Wiien used systemically, the ion is generally employed in an amount of from 1 to 10 mg. per kg. of host weight. Peptide or protein dosages may be within the ranges hereinabove described.

It is also to be understood that the peptide or protein and ion having pharmacological properties, may be delivered or administered in different forms; for example, the ion may be administered orally, while the peptide may be administered by IV or IP.

As representative examples of administering the peptide or protein and ion for topical or local administration, the peptide could be administered in an amount of up to about 1% weight to weight and the ion delivered in an amount of about 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 or protein. For example, the peptide or protein 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 or proteins of the present invention may be administered to a SUBSTITUTE SHEET (RULE 28) WO 95/19370 40 PCT/US95/00714 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 gentamicin C 1 gentamicin C 2 gentamicin Cla), netilmicin, 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 6-0-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 11

/C

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 SUBSTITUTE SHEET (RULE 28) I WO 95/19370 PCT/UIS95/00714 41 Brewery); a 6-0-methyl-l-4"-0-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 moiety, such as 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 or protein and antibiotic may be administered by direct administration to a target cell or by systemic or tropical 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 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 or protein dosages may be those as hereinabove described.

As representative examples of administering the peptide or protein and antibiotic for topical or local administration, the peptide or protein could be administered in an amount of from about 0.1% to about 10% weight to weight, and the antibiotic is delivered in an amount of from about 0.1% to about 10% weight to weight.

SUBSTITUTE SHEET (RULE 28) WO 95/19370 PCTIUS95/00714 42 In accordance with another embodiment, the peptides or proteins 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 or proteins 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.

The present invention will be further described with respect to the following examples; however, the scope of the invention is not to be limited thereby.

EXAMPLE 1 Table I, which follows, indicates the Minimal Inhibitory Concentration (MIC) in Ag/ml of various peptides, against S.aureus strain ATCC 25923(S), P. aeruqinosa strain ATCC 27853(P), and E. coli ATCC strain 25922(E), and C.albicans A indicates that each amino acid residue is a D-amino acid residue or a glycine residue. The peptides are SUBSTITUTE SHEET (RULE 28) Wd 95/19370 PCT/US95/n0714 43 unsubstituted at the N-terminal, substituted with an acetyl group at the N-terminal as indicated by Ac-; substituted with an octanoyl group at the N-terminal as indicated by Oct-, substituted with sphingosine as indicated by Sph-; substituted with a succinyl group, as indicated by Suc-; substituted with a hexanoyl group, as indicated by Hex-; substituted with a heptanoyl group, as indicated by Hep-; substituted with a valeryl group, as indicated by Val-; substituted with a myristryl group, as indicated by Myr-; or substituted with an ibuprofyl group, as indicated by Ibu-.

The procedure for the antibacterial assay is based upon the guidelines of the National Committee for Clinical Laboratory Standards, Document M7-T2, Volume 8, No. 8, 1988.

Stock solutions of peptides with and without the appropriate substitutions, are prepared at a concentration 512 Ag/ml in sterile deionized distilled water and stored at 0 C. Each peptide is a C-terminal amide.

The stock peptide solution is diluted in serial dilutions down the wells of a microtiter plate so that the final concentrations of peptides in the wells are 0.25, 0.50, 1, 2, 4, 8, 16, 32, 64, 128, and 256 pg/ml. 1-5 X 105 CFUs/ml of either S.aureus ATCC 25923, E. coli ATCC 25922, P.

aeruginosa ATCC 27853, or C.albicans, were added to the wells in full strength Mueller Hinton broth (BBL 11443) from a mid-log culture. The inoculum is standardized spectrophotometrically at 600 nm and is verified by colony counts. The plates are incubated for 16-20 hours at 37 0

C,

and the minimal inhibitory concentration (MIC) for each peptide is determined. Minimal inhibitory concentration is defined as the lowest concentration of peptide which produces a clear well in the microtiter plate. The minimal inhibitory concentration of each of the peptides with and/or without the appropriate substitutions is given in Table I below.

SUBSTITUTE SHEET (RULE 28) WO'95/19370 PCT/US95/00714 44 Table 1 Minimal Inhibitory Concentration ug/ml) Peptide S P E CA Oct-(SEQ ID NO: 27)-NH 2 2 4 2 16 Oct-(SEQ ID NO: 27)-OH 8 8 4 32 Ac-(SEQ ID NO: 27)-NH 2 32 128 8 N/A (SEQ ID NO: 27)-NH 2 8,16 64,128 8 N/A (SEQ ID NO: 27)-OH 128 128 8 N/A Sph-Suc-(SEQ ID NO: 27)-NH 2 64 >256 32 N/A Suc-(SEQ ID NO: 27)-NH 2 >256 >256 32 128 Ibu-(SEQ ID NO: 27)-NH 2 2 4 8 128 (SEQ ID NO: 66)-NH 2 4 32 32 64 Oct-(SEQ ID NO: 66)-NH 2 4 16 8 256 (SEQ ID NO: 86)-OH 128 32 2 256 Oct-(SEQ ID NO: 86)-OH 8 4 2 128 Oct-(SEQ ID NO: -NH 2 128 32,64 128 64,128 Oct-(SEQ ID NO: 107)-NH 2 128 256 >256 128 Oct-(SEQ ID NO: 108)-NH 2 16 4 64 64 Oct-(SEQ ID NO: 109)-NH 2 8 4 16 32 (SEQ ID NO: 110)-NH 2 >256 32,64 64,128 N/A Ac-(SEQ ID NO: 110)-NH 2 256 8,16 32,64 N/A Oct-(SEQ ID NO: 110)-NH 2 4 4 8 32 Oct-D-(SEQ ID NO: 110)-NH 2 4 4 16 32 Hex-(SEQ ID NO: 110)-NH 2 16 8 16 64 Hep-(SEQ ID NO: 110)-NH 2 8 4 16 32 Val-(SEQ-ID NO: 110)-NH 2 64 8 32 32 Myr-(SEQ ID NO: 110)-NH 2 16 16 16 >256 Oct-(SEQ ID NO: 111)-NH 2 64 8 32 32 (SEQ ID NO: 113)-NH 2 16,32 8,16 32 N/A Ac-(SEQ ID NO: 113)-NH 2 32 64 64 N/A Oct-(SEQ ID NO: 113)-NH 2 8 8 8 128 Oct-(SEQ ID NO: 118)-NH 2 >256 256 >256 >256 Oct-(SEQ ID NO: 119)-NH 2 >256 >256 >256 >256 SUBSTITUTE SHEET (RULE 28) W6 95/19370 45 PCT/US95/00714 Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ Oct- (SEQ_ Oct- (SEQ

(SEQ

Oct- (SEQ Hex- (SEQ Myr- (SEQ Oct- (SEQ 120)

-NH

2 121)

-NH

2 122)

-NH

2 123)

-NH

2 124)

-NH

2 125)

-NH

2 126)

-NH

127)

-NH

128)

-NH

2 129)

-NH

130)

-NH

2 131)

-NH

2 132) -NH2 133)

-NH

2 134) -NH2 135)

-NH

2 136) -NH 2 137)

-NH

2 138)

-NH

2 139)

-NH

2 140)

-NH

2 141)

-NH

2 142)

-NH

2 143)

-NH

144)

-NH

2 145)

-NH

2 146)

-NH

2 147)

-NH

2 148)

-NH

2 149)

-NH

2 149)

-NH

2 150)

-NH

2 151)

-NH

2 153)

-NH

2 64 128 32 32 128 8 8 >256 128 128 4 8 32 16 64 32 256 256 4 16 32 4 4 16 8 4 8 32 32 256 64 16 61 8 128 256 32 16 64 8 8 32 64 16 4 8 8 32 8 8 64 256 8 32 8 4 2 2 4 8 4 32 8 32 16 128 128 8 256 256 64 32 256 16 8 32 32 32 4 4 8 32 16 64 256 >256 8 16 16 8 8 16 16 16 16 32 32 32 32 32 64 32 64 256 64 32 128 64 64 128 32 128 8 64 64 128 64 64 128 256 64 128 64 32 32 16 32 64 32 128 128 64 128 128 >256 64 SUBSTITUTE SHEET (RULE 28) WO 95/19370 PCT/US95/00714 46 The above results indicate that when a biologically active peptide is substituted with a lipophilic moiety of the present invention, the peptide has increase biological activity against a variety of microorganisms.

EXAMPLE 2 Stock cultures of P. qinqivalis, S. mutans or A.

viscosus are maintained on Brucella blood agar plates with hemin and vitamin K 1 (BBL, Cockeysville, MD) and are grown under anaerobic conditions (Coy Anaerobic Chamber, Ann Arbor, MI) with an atmosphere of 80% N2-10%H2-1-%CO 2 at 370C.

Experimental cultures are grown up in Brain heart infusion (BHI) broth, (BBL, Cockeysville, MD) plus hemin mg/liter) (Sigma Chemical Co., St. Louis, IL) plus vitamin K1 (0.25 mg/liter) (Sigma Chemical Co., St. Louis, MO). For susceptibility testing cultures'are taken from overnight (24 hour) broth cultures and diluted in fresh BHI broth (plus hemin plus vitamin K to deliver 1 x 106 colony-forming units (CFUs)/ml in each microtiter test well.

Antimicrobial susceptibility tests are performed according to the guidelines of the National Committee for Clinical Laboratory Standards (NCCLS) (Document M11-T2, 1989). Microtiter plates (Corning, Corning, NY) are filled aseptically with BHI broth (plus hemin plus vitamin K 1 to a volume of 100 Al by the use of a Beckman Biomek 1000 robotic instrument (Beckman Instruments, Palo Alto, CA). Peptides are tested in duplicate lanes by adding manually 100 Al of a 1.024 mg/ml peptide solution in water to the top wells of a microtiter plate lane. The peptide is diluted serially 1:2 by mixing and transferring 100 Al from the top well down to the bottom well in the lane by use of the Beckman Biomek 1000 (Beckman Instruments, Palo Alto, CA). The last 100 il from the bottom well is discarded. One hundred microliters of the bacterial are added in BHI (plus hemin plus vitamin K to each test well to give final peptide dilutions from 0.25 Ag/ml. The plates are incubated in the anaerobic SUBSTITUTE SHEET (RULE 28) Wd 95/19370 PCT/US95/00714 47 chamber at 37 0 C for 24-48 hours. After incubation, the minimum inhibitory concentration (MIC) is determined as the lowest concentration of peptide which inhibits growth as determined by visual inspection and optical density when read on a Dynatech MR5000 microtiter plate reader at 630 nm (Dynatech Laboratories, Chantilly, VA). The results are given in Table II below.

Table II MIC (Al/ml) P. ginqivalis (strain) Peptide (SEQ ID NO: 27)-NH 2 Oct-(SEQ ID NO: 27)-NH 2 Oct-(SEQ ID NO: 27)-OH (SEQ ID NO: 66)-NH 2 Oct-(SEQ ID NO: 66)-NH 2 (SEQ ID NO: 86)-NH 2 (SEQ ID NO: 86)-OH A7A1 381 128 16 2 16 4 8 16

FAY-

-28 16 4 1 4 8 9- 19M-1 64 4 2 8 2 14K-1 W50 8,128 4 4 16 S mutans A.vis'cos 16 16 16 16 2 1 16 2 4 8 2 2 32 8 4 4 16 4 16 2 8,128 4 8 Oc:t-(SEQ ID NO: 86)-OH 1 1 1 1 1 16 16 (SEA ID NO: 152)-NH 2 32 4,8 8 4 4 16 16 Example 3 CD-1 male mice (average body weight, 22.8g) were inoculated with live E.coli strain 21915-1 (2.3 x 105 CFU/ mouse) by injection intraperitoneally. Oct.-(SEQ ID NO:143)-NH 2 then was injected intravenously via the tail vein at 1 and 5 hours post-inoculation. Control mice were inoculated and treated with 0.9% saline. Each different treatment group had 10 mice per group. All control mice died. Treatment doses of Oct-(SEQ ID No:143)-NH 2 were 1, and 20 mg/kg in toto, and resulted in 20%, 40%, 90%, and survival at six days post-inoculation, respectively.

SUBSTITUTE SHEET (RULE 28) I W 95/19370 PCT/US95/00714 48 Example 4 Oct-(SEQ ID NO:143)-NM} 2 was injected intravenously into male C57BL/6J mice (average body weight, 20.1g) approximately two minutes prior to intraperitoneal injection of a solution of lipopolysaccharide (either 0.1 Ag or 0.5 Ag mouse) from E.coli serotype 0111:B4 and galactosamine (8 mg/mouse).

Treatment doses of Oct.-(SEQ ID NO:143)-NH 2 were 0, 5, 12.5, or 15 mg/kg (10 mice/group), and when administered prior to 0.5 tg lipopolysaccharide/mouse resulted in 30%, 50%, and 60% survival at five days post-lipopolysaccharide administration, respectively. When these doses were administered prior to the administration of 0.1 Ag lipopolysaccharide/mouse, 7he results were 40%, 100%, 100%, 100%, and 100% survival at five days post-lipopolysaccharide administration, respectively.

Example A stock solution (10x) of 0.6 mM dye is prepared by adding 1.68 mg of (1-ethyl-2-(3-[1-ethylnapthol(1,2-d)thiazolin-2- ylidene]-2-methylpropenyl)naphtho-(1,2-d)thiazolium bromide (Signa E-7762) to 5 ml of 200 proof ethanol. 1 ml of this solution was added to 9 ml ethanol to give 0.06 mM of dye (60 AM dye).

A stock solution of lipopolysaccharide (LPS) from E.coli serotype 0111:B4 was prepared at 1.5 mg/ml. 400 Al of this solution was mixed with 4.6 ml pyrogen free water to give a 120 g/ml solution.

Row 1 and rows 3 through 12 of a microtiter plate were filled with 100 pl of pyrogen free water or with 10 mg/ml of bovine serum albumin. 200 p1 of peptide then is added to row 2 of the microtiter plate at a concentraiton of 1 ml/ml.

200 Al of pyrogen free water is added to each of the control wells in two lanes (having dye and LPS but no peptide or having dye and no LPS and no peptide). 100 pl then is serially diluted from row 2 through row 12 of the microtiter plate. 50 l of PBS (pH 7.4) and 50 1 of the LPS solution then are added to row 1 of the plate (blank wells).

SUBSTITUTE SHEET (RULE 28) WO95/19370 PCT/US95/00714 49 Equal volumes of the LPS solution, the dye, and PBS (pH 7.4, approx. 150 mM) are mixed to form a dye-buffer LPS mixture having LPS at a final concentration of 20 JM. The dye-buffer LPS mixture then is incubated for 10 minutes at room temperature in the dark.

100 6g1 of the dye-LPS-buffer mixture then is added to every well of the microtiter plate except to the blank wells and to the control lane that does not have LPS or peptide.

The plate is incubated for 10 minutes at room temperature in the dark and the absorbance at 460 nm and 510 nm is read.

From these absorbances, the LPS50 value, which is the concentration in Ag/ml of peptide necessary to inhibit the binding of 50% of the lipopolysaccharide to the dye, is calculated.

The above procedure was carried out for the peptides listed in Table III below.

Table III Peptide Oct-(SEQ ID NO:106) Oct-(SEQ ID NO:107) Oct-(SEQ ID NO:109) Oct-(SEQ ID NO:110) Oct-D-(SEQ ID NO:110)- Oct-D-(SEQ ID NO:110)- Oct-(SEQ ID NO:111) Oct-(SEQ ID NO:121) Oct-(SEQ ID NO:123) Oct-(SEQ ID NO:137) Oct-(SEQ ID NO:138) Oct-(SEQ ID NO:142) Oct-(SEQ ID NO:143) LPS50 (uc/ml

NH

2 6.80

NH

2 15.00 NH2 0

NH

2 0.84

NH

2 0.97

NH

2 -melibionic acid 45.00

NH

2 1.00

NH

2 20.00

NH

2 1.70

NH

2 4.80

NH

2 1.00

NH

2 0.70

NH

2 0.90 The peptides or proteins of the present invention, whether administered alone or in combination with agents such as ions having pharmacological properties, antibiotics, or other biologically active peptides or proteins as hereinabove described, may be employed in a wide variety of SUBSTITUTE SHEET (RULE 26) W6 95119370 prT/Toqll/nm i,1 pharmaceutical compositions in combination with a non-toxic pharmaceutical carrier or vehicle such as a f:i.ller, 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 peptides or proteins and/or agent as hereinabove described may alsQ 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, parasites, fungi, and the like.

The peptides or proteins may be administered to a host in particular an animal, in an effective antibiotic and/or anti-tumor and/or antiviral and/or antimicrobial and/or antispermicidal and/or antifungal and/or antiparasitic amount, or in an amount effective to stimulate wound healing in a host, or in an amount effective in treating septic shock in a host. The peptides or proteins may be administered either alone or in combination with an ion having pharmacological properties, antibiotic, or ion channel forming peptide or protein as hereinabove described. When the peptide or protein is administered in combination with an ion having pharmacological properties, the activity of the peptide or protein is potentiated.

When the peptide or protein is administered in combination with an agent as hereinabove described, it is possible to administer the peptide and agent in separate forms. For example, the agent may be administered systemically and the peptide or protein may be administered topically.

When the peptide or protein is administered topically, it may be administered in combination with a water-soluble vehicle, said water-soluble vehicle being in the form of an ointment, cream, lotion paste or the like. Examples of water-soluble vehicles which may be employed include, but are SUBSTITUTE SHEET (RULE 28) wi o4/1 0n7n0 T^lr'rrvlcnr )Ilhl1i .d 51 Jr% U0F0IU U not limited to, glycols, such as polyethylene glycol, hydroxycellulose, and KY Jelly. The water-soluble vehicle is preferably free of an oil substance.

The peptide or protein may also be employed alone, or in combination with an ion having pharmacological properties, as hereinabove described in the form of an oral composition for oral hygiene. Such a composition may be incorporated into a wide variety of compositions and materials used for oral hygiene purposes, which include, but are not limited to, toothpastes, mouthwashes, tooth gels, and tooth powders.

Such composition may thus be used to treat or prevent periodontal disease, to prevent or reduce plaque, gingivitis, and/or to prevent or treat or reduce dental caries. The peptide and ion having pharmacological properties may be used to inhibit, prevent, or destroy the growth of Streptococcus mutans, which is associated with dental caries and periodontal disease.

It is to be understood, however, that the scope of the present invention is not to be limited to the specific embodiments described above. The invention may be practiced other than as particularly described and still be within the scope of the accompanying claims.

SEQUENCE LISTING GENERAL INFORMATION: APPLICANT: Kari, U. Prasad (ii) TITLE OF INVENTION: Biology Active Peptides Having N-Terminal Substitutions (iii) NUMBER OF SEQUENCES: 153 (iv) CORRESPONDENCE ADDRESS: ADDRESSEE: Carella, Byrne, Bain, Gilfillan, Cecchi Stewart STREET: 6 Becker Farm Road CITY: Roseland STATE: New Jersey COUNTRY: USA ZIP: 07068 SUBSTITUTE SHEET (RULE 28) Wd 95/19370 52 PCT/US95/00714 COMPUTER READABLE FORM: MEDIUM TYPE: 3.5 inch diskette COMPUTER: IBM PS/2 OPERATING SYSTEM: PC-DOS SOFTWARE: DW4.V2 (vi) CURRENT APPLICATION DATA: APPLICATION NUMBER: FILING DATE:

CLASSIFICATION:

(vii) PRIOR APPLICATION DATA: APPLICATION NUWBER: 07/891,201 FILING DATE: 01-JUN-1992 (viii) ATTORNEY/AGENT INFORMATION: NAME: Olstein, Elliott M.

REGISTRATION NUMBER: 24,025 REFERENCE/DOCKET NUMBER: 421250 (ix) TELECOMMUNICATION INFORMATION: TELEPHONE: 201-994-1700 TELEFAX: 201-994-1744 INFORMATION FOR SEQ ID NO:1: SEQUENCE CHARACTERISTICS: LENGTH: 20 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide PUBLICATION INFORMATION: DOCUMENT NUMBER: W089/11290 FILING DATE: 19-MAY-1989 PUBLICATION DATE: 30-NOV-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys SUBSTITUTE SHEET (RULE 28) WO 95/19370 53 INFORMATION FOR SEQ ID NO:2: SEQUENCE CHARACTERISTICS: LENGTH: 24 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide PUBLICATION INFORMATION: DOCUMENT NUMBER: W089/11290 FILING DATE: 19-MAY-1989 PUBLICATION DATE: 30-NOV-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys INFORMATION FOR SEQ ID NO:3: SEQUENCE CHARACTERISTICS: LENGTH: 16 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide PUBLICATION INFORMATION: DOCUMENT NUMBER: W089/11290 FILING DATE: 19-MAY-1989 PUBLICATION DATE: 30-NOV-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala PCT/US95/00714 SUBSTITUT'E SHEET (RULE 28) 4 W695/19370 54 PCT/US95/00714 INFORMATION FOR SEQ ID NO:4: SEQUENCE CHARACTERISTICS: LENGTH: 20 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide PUBLICATION INFORMATION: DOCUMENT NUMBER: W089/11290 FILING DATE: 19-MAY-1989 PUBLICATION DATE: 30-NOV-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 16 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide PUBLICATION INFORMATION: DOCUMENT NUMBER: W089/11290 FILING DATE: 19-MAY-1989 PUBLICATION DATE: 30-NOV-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser INFORMATION FOR SEQ ID NO:6: SEQUENCE CHARACTERISTICS: LENGTH: 23 amino acids TYPE: amino acid SUBSTITUTE SHEET (RULE 26) W6 95/19370 55

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Magainin I peptide.

PUBLICATION INFORMATION: AUTHOR: Zasloff, Michael JOURNAL: Proc. Nat. Acad. Sci.

VOLUME: 84 PAGES: 5449-5453 DATE: AUG 1987 DOCUMENT NUMBER: US 4810777 FILING DATE: 04-MAR-1987 PUBLICATION DATE: 07-MAR-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Gly Ile Gly Lys Phe Leu His Ser Ala Gly Lys Phe Gly Lys Ala Phe Val Gly Glu Ile Met Lys Ser INFORMATION FOR SEQ ID NO:7: SEQUENCE CHARACTERISTICS: LENGTH: 23 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Magainin II peptide.

VOLUME: 84 PAGES: 5449-5453 DATE: AUG 1987 DOCUMENT NUMBER: US 4810777 PCT/US9S/00714 SUBSTITUTE SHEET (RULE Wd 95/19370 5 56 FILING DATE: 04-MAR-1987 PUBLICATION DATE: 07-MAR-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: Gly Ile Gly Lys Phe Leu His Ser Ala Lys Lys Phe Gly Lys Ala Phe Val Gly Glu lie Met Asn Ser INFORMATION FOR SEQ ID NO:8: SEQUENCE CHARACTERISTICS: LENGTH: 22 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: Magainin III peptide.

VOLUME: 84 PAGES: 5449-5453 DATE: AUG 1987 DOCUMENT NUMBER: US 4810777 FILING DATE: 04-MAR-1987 PUBLICATION DATE: 07-MAR-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: Gly Ile Gly Lys Phe Leu His Ser Ala Lys Lys Phe Gly Lys Ala Phe Val Gly Glu Ile Met Asn INFORMATION FOR SEQ ID NO:9: SEQUENCE CHARACTERISTICS: LENGTH: 22 amino acids TYPE: amino acid PCT/US95/00714 SUBSTITUTE SHEET (RULE 26) WO 95/19370 57 57

STRANDLPNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: magainin peptide.

VOLUME: 84 PAGES: 5449-5453 DATE: AUG 1987 DOCUMENT NUMBER: US 4810777 FILING DATE: 04-MAR-1987 PUBLICATION DATE: 07-MAR-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: Ile Gly Lys Phe Leu His Ser Ala Lys Lys Phe Gly Lys Ala Phe Val Gly Glu Ile Met Asn Ser INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORPMATION: AUTHOR: Zasloff, Michael JOURNAL: Proc. Nat. Acad. Sci.

VOLUME: 84 PAGES: 5449-5453 DATE: AUG 1987 DOCUMENT NUMBER: US 4810777 PCT/US95/00714 SUBSTITUTE SHEET (RULE 26) WO 95/19370 8 FILING DATE: 04-MAR-1987 PUBLICATION DATE: 07-MAR-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Lys Phe Leu His Ser Ala Lys Lys Phe Gly Lys Ala Phe Val Gly Glu Ile Met Asn Ser INFORMATION FOR SEQ ID NO:11: SEQUENCE CHARACTERISTICS: LENGTH: 20 amino acids TYPE: amino acid

STRANDEDNESS:

VOLUME: 84 PAGES: 5449-5453 DATE: AUG 1987 DOCUMENT NUMBER: US 4810777 FILING DATE: 04-MAR-1987 PUBLICATION DATE: 07-MAR-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: Lys Phe Leu His Ser Ala Lys Lys Phe Gly Lys Ala Phe Val Gly Glu Ile Met Asn Ser INFORMATION FOR SEQ ID NO:12: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

PCT/US95/00714 SUBSTITUTE SHEET (RULE 28) SWO 95/19370 59 PCT/US95/00714 TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: PGLa peptide.

PUBLICATION INFORMATION: AUTHOR: Hoffman, et al.

JOURNAL: EMBO J.

VOLUME: 2 PAGES: 711-714 DATE: 1983 AUTHOR: Andreu, et al.

JOURNAL: Journal of Biochemistry VOLUME: 149 PAGES: 531-535 DATE: 1985 AUTHOR: Gibson, et al.

JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 5341-5349 DATE: 1986 AUTHOR: Giovannini, et al.

JOURNAL: Biochem J.

VOLUME: 243 PAGES: 113-120 DATE: 1987 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: Gly Met Ala Ser Lys Ala Gly Ala Ile Ala Gly Lys Ile Ala Lys Val Ala Leu Lys Ala Leu INFORMATION FOR SEQ ID NO:13: SEQUENCE CHARACTERISTICS: LENGTH: 25 amino acids TYPE: amino acid SUBSTITUTE SHEET (RULE 28) WO 95/19370

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: XPF peptide.

PUBLICATION INFORMATION: AUTHOR: Hoffman, et al.

JOURNAL: EMBO J.

VOLUME: 2 PAGES: 711-714 DATE: 1983 AUTHOR: Andreu, et al.

JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 5341-5349 DATE: 1986 AUTHOR: Giovannini, et al.

JOURNAL: Biochem J.

VOLUME: 243 PAGES: 113-120 DATE: 1987 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: Gly Trp Ala Ser Lys Ile Gly Gin Thr Leu Gly Lys Ile Ala Lys Val Gly Leu Lys Glu Leu Ile Gin Pro Lys INFORMATION FOR SEQ ID NO:14: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids PCTIUS95/00714 SUBSTITUTE SHEET (RULE 26) WVO95119370 61 TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: CPF peptide.

CW PUBLICATION INFORMATION: AUTHOR: Ri4chter, K.

Egger-, R.

Kre il JOURNAL: J. Biol. Chem.

PC'1rUS95A)0714

(D)

(F)

(G)

(A)

(C)

CD)

(F)

(G)

(A)

(C)

(D)

(F)

CG)

VOLUME:

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261 3676-3680 1986 Wakabayashi, T.

Kato, H.

Tachibaba, S.

Nucleic Acids Research 13 1817-1828 1985 Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

J. Biol. Chem.

261 5341- 534 9 1986 DOCUMENT NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: Gly Phe Gly Ser Phe Leu Gly Leu Ala Leu SUBSTITUTE SHEET (RULE 28) I W695/19370 62 PCT/US95/00714 Lys Ala Ala Leu Lys Ile Gly Ala Asn Ala Leu Gly Gly Ala Pro Gin Gin INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

VOLUME:

PAGES:

DATE:

AUTHOR:

JOURNAL:

VOLUME:

PAGES:

DATE:

AUTHOR:

JOURNAL:

VOLUME:

PAGES

DATE:

261 3676-3680 1986 'Wakabayashi, T.

Kato, H.

Tachibaba, S.

Nucleic Acids Research 13 1817-1828 1985 Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

J. Biol. Chem.

261 5341-5349 1986 SUBSTITUTE SHEET (RULE 28) W6 95/19370 63 DOCUMENT NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Leu Ala Ser Phe Leu Gly Lys Ala Leu Lys Ala Gly Leu Lys Ile Gly Ala His Leu Leu Gly Gly Ala Pro Gln Gin INFORMATION FOR SEQ ID NO:16: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

PCT/US95/00714

VOLUME:

PAGES:

DATE:

AUTHOR:

JOURNAL:

VOLUME:

PAGES:

DATE:

AUTHOR:

261 3676-3680 1986 Wakabayashi, T.

Kato, H.

Tachibaba, S.

Nucleic Acids Research 13 1817-1828 1985 Gibson, B.W.

Poulter, L.

SUBSTITUTE SHELi 1tLE 28) WO 95/19370 64 PCT/US95/00714 Williams, D.H.

Maggio, J.E.

JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 5341-5349 DATE: 1986 DOCUMENT NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: Gly Leu Ala Ser Leu Leu Gly Lys Ala Leu Lys Ala Gly Leu Lys Ile Gly Thr His Phe Leu Gly Gly Ala Pro Gln Gin INFORMATION FOR SEQ ID NO:17: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 3676-3680 DATE: 1986 AUTHOR: Wakabayashi, T.

Kato, H.

Tachibaba, S.

SUBSTITUTE SHEET (RULE 28) WO 9)5/19)370 PCT/US95/00714 65 JOURNAL: Nucleic Acids Research VOLUME: 13 PAGES: 1817-1828 DATE: 1985 AUTHOR: Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 5341-5349 DATE: 1986 DOCUMENT NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: Gly Leu Ala Ser Leu Leu Gly Lys Ala Leu Lys Ala Thr Leu Lys Ile Gly Thr His Phe Leu Gly Gly Ala Pro Gin Gln INFORMATION FOR SEQ ID NO:18: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil SUBSTITUTE SHEET (RULE 28) WO 95/19370 66 WO 9519370PCTIUS95/007 14

(C)

(D)

(F)

(G)

(A)

(C)

(D)

(F)

(G)

(A)

(C)

(D)

(F)

(G)

(H)

JOURNAL:

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Biol. Chem.

261 3676-3680 1986 Wakabayashi, T.

Kato, H.

Tachibaba, S.

Miriciir Ae-iri R esearch 13 1817-1828 1985 Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

Biol. Chem.

261 5341-5349 1986 NUMBER: W090/04407 FILING DATE: 16-OCT--1989 PUBLICATIOIN DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: Gly Phe Ala Ser Phe Leu Gly Lys Ala Leu Lys Ala Ala Leu Lys Ile Gly Ala Asn Met Leu Gly Gly Thr Pro Gln Gln INFORMATION FOR SEQ ID NO:19: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide SUBSTITUTE SHEET (RULE 28) Wd 95/19370 PCIA)S95/00714 67 (ix) FEATURE: NAME/KEY: CPF peptide.

(x PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: 1. Biol. Chem.

VOLUME: 262.

PAGES: 3676-3680 DATE: 1986

AUTHOR:

JOURNAL:

VOLUME:

PAGES:

DATE:

AUTHOR:

JOURNAL:

VOLUME:

PAGES:

DATE:

DOCUMENT

Wakabayashi, T.

Kato, H.

Tachibaba, S.

Nucleic Acids Research 13 1817-1828 1985 Gibson, 8.W.

Poulter, L.

Williamas, D.H.

Maggio, J.E.

J. Biol. Chem.

261 5341- 534 9 1986 NUMBER: W090/04407 FILING DATE: 16-OCT-1989 (JI) PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:i9: Gly Phe Gly Ser Phe Leu Gly Lys Ala Leu Lys Ala Ala Leu Lys Ile Gly Ala Asn Ala Leu Gly Gly Ala Pro Gin Gin SUB3STITUTE SHEET (RULE 26) W6 95/19370 68 PCT/US95/00714 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

(D)

(F)

(G)

(A)

(C)

(D)

(F)

(G)

(A)

(C)

(D)

(F)

(G)

(H)

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DOCUMENT

261 3676-3680 1986 Wakabayashi, T.

Kato, H.

Tachibaba, S.

Nucleic Acids Research 13 1817-1828 1985 Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

J. Biol. Chem.

261 5341-5349 1986 NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 SUBSTITUTE SHEET (RULE 28) WO 95/19370 69 (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Phe Gly Ser Phe Leu Gly Lys Ala Leu Lys Ala Ala Leu Lys Ile Gly Ala Asn Ala Leu Gly Gly Ser Pro Gln Gin INFORMATION FOR SEQ ID NO:21: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: NAME/KEY: CL. peptide.

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 3676-3680 DATE: 1986 AUTHOR: Wakabayashi, T.

Kato, H.

PCT/US95/00714

JOURNAL:

VOLUME:

PAGES:

DATE:

AUTHOR:

JOURNAL:

Tachibaba, S.

Nucleic Acids Research 13 1817-1828 1985 Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

J. Biol. Chem.

SUBSTITUTE SHEET (RULE 26) WO 95/19370 70 PCT/US95/00714 VOLUME: 261 PAGES: 5341-5349 DATE: 1986 DOCUMENT NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: Gly Phe Ala Ser Phe Leu Gly Lys Ala Leu Lys Ala Ala Leu Lys Ile Gly Ala Asn Leu Leu Gly Gly Thr Pro Gln Gln INFORMATION FOR SEQ ID NO:22: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 3676-3680 DATE: 1986 AUTHOR: Wakabayashi, T.

Kato, H.

Tachibaba, S.

JOURNAL: Nucleic Acids Research VOLUME: 13 PAGES: 1817-1828 SUBSTITUTE SHEET (RULE 26) WO 95/19370 71 PCT/US95/00714 DATE: 1985 AUTHOR: Gibson, B.N.

Poulter, L.

Williams, D.H.

Maggio, J.E.

JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 5341-5349 DATE: 1986 DOCUMENT NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: Gly Phe Ala Ser Phe Leu Gly Lys Ala Leu Lys Ala Ala Leu Lys Ile Gly Ala Asn Ala Leu Gly Gly Ala Pro Gin Gin INFORMATION FOR SEQ ID NO:23: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 3676-3680 DATE: 1986 SUBSTITUTE SHEET (RULE 28) I WO 95/19370 72 PCT/US95/00714 AUTHOR: Wakabayashi, T.

Kato, H.

Tachibaba, S.

JOURNAL:

VOLUME:

PAGES:

DATE:

AUTHOR:

Nucleic Acids Research 13 1817-1828 1985 Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 5341-5349 DATE: 1986 DOCUMENT NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: Gly Phe Ala Ser Phe Leu Gly Lys Ala Leu Lys Ala Ala Leu Lys Ile Gly Ala Asn Met Leu Gly Gly Ala 2ro Gln Gln INFORMATION FOR SEQ ID NO:24: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

SUBSTITUTE SHEET (RULE 26) WO 95/19370 73 PCT/US95/007 14 PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 3676-3680 DATE: 1986 AUTHOR: Wakabayashi, T.

Kato, H.

Tachibaba, S.

JOURNAL: Nucleic Acids Research

VOLUME:

PAGES:

DATE:

AUTHOR:

13 1817-1828 1985 Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

J. Biol. Chem.

261 5341-5349 1986 NUMBER: W090/04407

JOURNAL:

VOLUME:

PAGES:

DATE:

DOCUMENT

FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: Gly Phe Gly Ser Phe Leu Gly Lys Ala Leu Lys Ala Ala Leu Lys Ile Gly Ala Asn Ala Leu Gly Gly Ser Leu Gin Gin INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids SUBSTITUTE SHEET (RULE 28) WO 95/19370 74 P'CT/US95/00714 TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 3676-3680 DATE: 1986 AUTHOR: Wakabayashi, T.

Kato, H.

Tachibaba, S.

JOURNAL: Nucleic Acids Research VOLUME: 13 PAGES: 1817-1828 DATE: 1985 AUTHOR: Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 5341-5349 DATE: 1986 DOCUMENT NUMBER: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Phe Gly Ser Phe Leu Gly Lys Ala Leu SUBSTITUTE SHEET (RULE 26) WO 95/19370 75 Lys Ala Gly Leu Lys Ile Gly Thr Asn Phe PCT/US95/00714 Leu Gly Gly Ala Pro Gin Gin (2) INFORMATION FOR SEQ ID NO:26: SEQUENCE CHARACTERISTICS: LENGTH: 27 amino acids TYPE: amino acid

STRANDEDNESS:

PUBLICATION INFORMATION: AUTHOR: Richter, K.

Egger, R.

Kreil JOURNAL: J. Biol. Chem.

VOLUME: 261 PAGES: 3676-3680 DATE: 1986 AUTHOR: Wakabayashi, T

JOURNAL:

VOLUME:

PAGES:

DATE:

AUTHOR:

JOURNAL:

VOLUME:

PAGES:

DATE:

Kato, H.

Tachibaba, S.

Nucleic Acids Research 13 1817-1828 1985 Gibson, B.W.

Poulter, L.

Williams, D.H.

Maggio, J.E.

J. Biol. Chem.

261 5341-5349 1986 SUBSTITUTE SHEET (RULE 28) WO 95/19370 7PCT/US95/007 4 DOCUMENT NUME t: W090/04407 FILING DATE: 16-OCT-1989 PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: Gly Leu Ala Ser Leu Leu Gly Lys Ala Leu Lys Ala Ala Leu Lys Ile Gly Ala Asn Ala Leu Gly Gly Ser Pro Gin Gin INFORMATION FOR SEQ ID NO:27: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:28: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: S Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly SUBSTITUTE SHEET (RULE 28) WVO'95/19370 77 PCTIUS95/00714 INFORMATION FOR SEQ ID NO:29: SEQUENCE CHARACTERISTICS:: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: Lys Ile Ala Gly Lys Ile Gly Lys Ile Ala Gly Lys Ile Gly Lys Ile Ala Gly Lys Ile Gly INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Lys Leu Ala Gly Lys Leu Ala Lys Leu Ala Gly Lys Leu Ala Lys Leu Ala Gly Lys Leu Ala INFORMATION FOR SEQ ID NO:31 SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: Lys Phe Ala Gly Lys Phe Ala Lys Phe Ala SUBSTITUTE SHEET (RULE 28) sB Pe WO 95/19370 78 PC$T/US95/00714 Gly Lys Phe Ala Lys Phe Ala Gly Lys Phe Ala INFORMATION FOR SEQ ID NO:32: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: Lys Ala Leu Ser Lys Ala Leu Lys Ala Leu Ser Lys Ala Leu Lys Ala Leu Ser Lys Ala Leu INFORMATION FOR SEQ ID NO:33: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: Lys Leu Leu Lys Ala Leu Gly Lys Leu Leu Lys Ala Leu Gly Lys Leu Leu Lys Ala Leu Gly INFORMATION FOR SEQ ID NO:34: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide SUBSTITUTE SHEET (RULE 26) is W 95/19370 79 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: Lys Ala Ile Gly Lys Ala Ile Lys Ala Ile Gly Lys Ala Ile Lys Ala Ile Gly Lys Ala Ile INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Ile Ala Lys Ile Ala Lys Gly Ile Ala Lys Ile Ala Lys Gly Ile Ala Lys Ile Ala Lys INFORMATION FOR SEQ ID NO:36: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: Lys Ile Ala Lys Ile Phe Gly Lys Ile Ala Lys Ile Phe Gly Lys Ile Ala Lys Ile Phe Gly INFORMATION FOR SEQ ID NO:37: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid PCT/US95/00714 SUBSTITUTE SHEET (RULE 28)

II

SWO 95/19370 80 PCT/US95/00714

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: Gly Ile Ala Arg Ile Ala Lys Gly Ile Ala Arg Ile Ala Lys Gly Ile Ala Arg Ile Ala Lys INFORMATION FOR SEQ ID NO:38: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: S.Q ID NO:38: Lys Phe Ala Arg Ile Ala Gly Lys Phe Ala Arg Ile Ala Gly Lys Phe Ala Arg Ile Ala Gly INFORMATION FOR SEQ ID NO:39: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39: Gly Phe Ala Lys Ile Ala Lys Cly Phe Ala Lys Ile Ala Lys Gly Phe Ala Lys Ile Ala Lys SUBSTITUTE SHEET (RULE 28) I_ W urn 19n3( n n u jii- 81 r INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is ornithine (xi) SEQUENCE DESCRIPTION: SEQ ID Lys Ile Ala Gly Xaa Ile Ala Lys Ile Ala Gly Xaa Ile Ala Lys Ile Ala Gly Xaa Ile Ala INFORMATION FOR SEQ ID NO:41: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41: Lys Ile Ala Arg Ile Ala Gly Lys Ile Ala Arg Ile Ala Gly Lys Ile Ala Arg Ile Ala Gly INFORMATION FOR SEQ ID NO:42 SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) FEATURE: CT/US95/00714 SUBSTITUTE SHEET (RULE 28)

I

WO 95/19370 82 PCTUS95/00714 OTHER INFORMATION: Xaa is ornithine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42: Xaa Ile Ala Gly Lys Ile Ala Xaa Ile Ala Gly Lys Ile Ala Xaa Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:43 SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43: Gly Ile Ala Arg Ile Phe Lys Gly Ile Ala Arg Ile Phe Lys Gly Ile Ala Arg Ile Phe Lys INFORMATION FOR SEQ ID NO:44: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is norleucine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:44: Lys Xaa Ala Gly Lys Xaa Ala Lys Xaa Ala Gly Lys Xaa Ala Lys Xaa Ala Gly Lys Xaa Ala SUBSTITUTE SHEET (RULE 28) 9u/1 nO 21i n DN r/US95/00714 83 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID Lys Xaa Ala Gly Lys Ile Ala Lys Xaa Ala Gly Lys Ile Ala Lys Xaa Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:46: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:46: Lys Ile Ala Gly Lys Xaa Ala Lys Ile Ala Gly Lys Xaa Ala Lys Ile Ala Gly Lys Xaa Ala INFORMATION FOR SEQ ID NO:47: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear SUBSTITUTE SHEET (RULE 28) W lB' I 11I l1 nr T/US95/00714 WVJy/I1JIU 84 (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is norvaline.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:47: Lys Xaa Ala Gly Lys Xaa Ala Lys Xaa Ala Gly Lys Xaa Ala Lys Xaa Ala Gly Lys Xaa Ala INFORMATION FOR SEQ ID NO:48: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is norvaline.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:48: Lys Xaa Ala Gly Lys Ile Ala Lys Xaa Ala Gly Lys Ile Ala Lys Xaa Ala Gly Lys Xaa Ala INFORMATION FOR SEQ ID NO:49: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49: Lys Leu Leu Ser Lys Leu Gly Lys Leu Leu SUBSTITUTE SHEET (RULE 28) WO, 95/19370 85 PCTJUS95/00714 Ser Lys Leu Gly Lys Leu Leu Ser Lys Leu Gly INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Lys Leu Leu Ser Lys Phe Gly Lys Leu Leu Ser Lys Phe Gly Lys Leu Leu Ser Lys Phe Gly INFORMATION FOR SEQ ID NO:51: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOILCULE TYPE: peptide (ix) FEATRE: OTHER INFORMATION: Xaa is norvaline.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:51: Lys Ile Ala Gly Lys Xaa Ala Lys Ile Ala Gly Lys Xaa Ala Lys Ile Ala Gly Lys Xaa Ala INFORMATION FOR SEQ ID NO:52: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

SUBSTITUTE SHEET (RULE 28) WO 95/19370 86 TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52: His Ile Ala Gly His Ile Ala His Ile Ala Gly His Ile Ala His Ile Ala Gly His Ile Ala INFORMATION FOR SEQ ID NO:53: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53: Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile INFORMATION FOR SEQ ID NO:54: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54: Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys ICTrUS95/00714 SUBSTITUTE SHEET (RULE 28) I WO 95/19370 87 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Lys Ile Ala Gly Arg Ile Ala Lys Ile Ala Gly Arg Ile Ala Lys Ile Ala Gly Arg Ile Ala INFORMATION FOR SEQ ID NO:56: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56: Arg Ile Ala Gly Arg Ile Ala Arg Ile Ala Gly Arg Ile Ala Arg Ile Ala Gly Arg Ile Ala INFORMATION FOR SEQ ID NO:57: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57: Lys Val Ala Gly Lys Ile Ala Lys Val Ala PCT/US95/00714 SUBSTITUTE SHEET (RULE 28) WO 95/19370 8 PCT/US95/00714 Gly Lys Ile Ala Lys Val Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:58: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58: Lys Ile Ala Gly Lys Val Ala Lys Ile Ala Gly Lys Val Ala Lys Ile Ala Gly Lys Val Ala INFORMATION FOR SEQ ID NO:59: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59: Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide SUBSTITUTE SHEET (RULE 28) WO 95/19370 PCT/US95/00714 89 (ix) FEATURE: OTHER INFORMATION: Xaa is ornithine.

(xi) SEQUENCE DESCRIPTION: SEQ ID Xaa Ile Ala Gly Xaa Ile Ala Xaa Ile Ala Gly Xaa Ile Ala Xaa Ile Ala Gly Xaa Ile Ala INFORMATION FOR SEQ ID NO:61: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61: Lys Phe Ala Gly Lys Ile Ala Lys Phe Ala Gly Lys Ile Ala Lys Phe Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:62: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62: Lys Ile Ala Gly Lys Phe Ala Lys Ile Ala Gly Lys Phe Ala Lys Ile Ala Gly Lys Phe Ala SUBSTITUTE SHEET (RULE 28) WI o0/1t71'A? nD'T1Yn9 '5/nI71 A 90 INFORMATION FOR SEQ ID NO:63: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is cyclohexylalanine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:63: Lys Xaa Ala Gly Lys Ile Ala Lys Xaa Ala Gly Lys Ile Ala Lys Xaa Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:64: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:64: Lys Xaa Ala Lys Ile Ala Gly Lys Xaa Ala Lys Ile Ala Gly Lys Xaa Ala Lys Ile Ala Gly INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear ,1 SUBSTITUTE SHEET (RULE 28) Wrs 95/19037(1 rCT/i rS95/00714 91 (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Arg Ile Ala Gly Lys Ile Ala Arg Ile Ala Gly Lys Ile Ala Arg Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:66: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is homoarginine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:66: Xaa Ile Ala Gly Xaa Ile Ala Xaa Ile Ala Gly Xaa Ile Ala Xaa Ile Ala Gly Xaa Ile Ala INFORMATION FOR SEQ ID NO:67: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: Xaa is p-aminophenylalanine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:67: Xaa Ile Ala Gly Lys Ile Ala Xaa Ile Ala Gly Lys Ile Ala Xaa Ile Ala Gly Lys Ile Ala SUBSTITUTE SHEET (RULE 28) WO 95/19370 I 'CT/US95/00714 92 INFORMATION FOR SEQ ID NO:68: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: Xaa is p-aminophenylalanine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68: Lys Ile Ala Gly Xaa Ile Ala Lys Ile Ala Gly Xaa Ile Ala Lys Ile Ala Gly Xaa Ile Ala INFORMATION FOR SEQ ID NO:69: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69: Lys Leu Ala Ser Lys Ala Gly Lys Ile Ala Gly Lys Ile Ala Lys Val Ala Leu Lys Ala Leu INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is ornithine.

(xi) SEQUENCE DESCRIPTION: SEQ ID SUBSTITUTE SHEET (RULE 26) WO 95/19370 93 PCT/US95/00714 Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Xaa Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:71: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71: Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Arg Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:72: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:72: Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Xaa Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:73: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid SUBSTITUTE SHEET (RULE 26) I WAtO9/10 1l Pr T/US95/00714 VY S_ J~fl 94

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:73: Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Xaa Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:74: SEQUENCE CHARACTERISTICS: LENGTH.: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide

FEATURE:

OTHER INFORMATION: Xaa is ornithine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:74: Lys Phe Ala Gly Lys Phe Ala Lys Phe Ala Gly Xaa Phe Ala Lys Phe Ala Gly Lys Phe Ala INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID SUBSTITUTE SHEET (RULE 26) Wr 04/10o711 T) T I c ncnI I 1 A V V J t A- 95 i L-.I U .JI UI Lys Ile Ala Gly Lys Phe Ala Lys Ile Ala Gly Xaa Phe Ala Lys Ile Ala Gly Lys Phe Ala INFORMATION FOR SEQ ID NO:76: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa at residues 6, 13, and is norleucine; Xaa at residue residue 12 is ornithine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:76: Lys Ile Ala Gly Lys Xaa Ala Lys Ile Ala Gly Xaa Xaa Ala Lys Ile Ala Gly Lys Xaa Ala INFORMATION FOR SEQ ID NO:77: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77: Lys Met Ala Ser Lys Ala Gly Lys Ile Ala 14 SUBSTITUTE SHEET (RULE 28) WO 95/19370 96 PCT/US95/00714 Gly Lys Ile Ala Lys Val Ala Leu Lys Ala Leu INFORMATION FOR SEQ ID NO:78: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78: Lys Ile Ala Ser Lys Ala Gly Lys Ile Ala Gly Lys Ile Ala Lys Val Ala Leu Lys Ala Leu INFORMATION FOR SEQ ID NO:79: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:79: Lys Ile Ala Ser Lys Ala Gly Lys Xaa Ala Gly Lys Ile Ala Lys Val Ala Leu Lys Ala Leu INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

SUBSTITUTE SHEET (RULE 28) WO 95/19370 97 PCT/US95/00714 TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is norleucine.

(xi) SEQUENCE DESCRIPTION: SEQ ID Lys Leu Ala Ser Lys Ala Gly Lys Xaa Ala Gly Lys Ile Ala Lys Val Ala Leu Lys Ala Leu INFORMATION FOR SEQ ID NO:81: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:81: Lys Xaa Ala Ser Lys Ala Gly Lys Xaa Ala Gly Lys Ile Ala Lys Val Ala Leu Lys Ala Leu INFORMATION FOR SEQ ID NO:82: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is paminophenylalanine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:82: SUBSTITUTE SHEET (RULE 28) SWO 95/19370 98 Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Xaa Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:83: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83: Lys Ile Ala Gly Ala Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:84: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84: Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Ala Ile Ala Lys Ile Ala Gly Lys lie PCT/US95/00714 Ala (2) INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

SUBSTITUTE SHEET (RULE 28) WO 95/19370 99 'PCT/US95/00714 TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Ala Ile Ala INFORMATION FOR SEQ ID NO:86: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86: Lys Ile Ala Lys Lys Ile Ala Lys Ile Ala Lys Lys Ile Ala Lys Ile Ala Lys Lys Ile Ala INFORMATION FOR SEQ ID NO:87: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87: Lys Phe Ala Lys Lys Phe Ala Lys Phe Ala Lys Lys Phe Ala Lys Phe Ala Lys Lys Phe Ala SUBSTITUTE SHEET (RULE 268 WO 95/19370 100 INFORMATION FOR SEQ ID NO:88: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88: Lys Phe Ala Lys Lys Ile Ala Lys Phe Ala Lys Lys Ile Ala Lys Phe Ala Lys Lys Ile Ala INFORMATION FOR SEQ ID NO:89: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89: Ala Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Lys Ile Ala Gly Lys Ile Ala Ala Ile Ala PCT/US95/00714 SUBSTITUTE SHEET (RULE 28) WO 95/19370 101 PCT/US95/00714 Gly Lys Ile Ala Lys Ile Ala Gly Lys Ile Ala INFORMATION FOR SEQ ID NO:91: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Lys Ile Ala Gly Lys Ile Ala Lys Ile Al Gly Lys Ile Ala Ala Ile Ala Gly Lys Ile NO:91: e Ala INFORMATION FOR SEQ ID NO:92: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92: Gly Met Ala Ser Lys Ala Gly Lys Ile Ala Gly Lys Ile Ala Lys Val Ala Leu Lys Ala Leu (2) INFORMATION FOR SEQ ID NO:93: SEQUENCE CHARACTERISTICS: LENGTH: 26 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide SUBSTITUTE SHEET (RULE 216 WO 95/19370 102 PCT/US95/00714 (vi) ORIGINAL SOURCE ORGANISM: Apis mellifera (vii) FEATURE: NAME/KEY: melittin peptide PUBLICATION INFORMATION: AUTHORS: Habermann, E.

Jentsch, J.

TITLE: Sequenzanalyse des Melittins aus den tryptischen and peptischen Spaltstucken JOURNAL: Hoppe-Seyler's Zeitschri8ft Physiol. Chem VOLUME: 348 PAGES: 37-50 DATE: 1987 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93: Gly Ile Gly Ala Val Leu Lys Val Leu Thr Thr Gly Leu Pro Ala Leu Ile Ser Trp Ile Lys Arg Lys Arg Gin Gin INFORMATION FOR SEQ ID NO:94: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino .cid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94: Leu Lys Lys Leu Lys Lys Leu Leu Lys Leu Leu INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids SUBSTITUTE SHEET (RULE 28) WO 95/1937() 103 TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Leu Leu INFORMATION FOR SEQ ID NO:96: SEQUENCE CHARACTERISTICS: LENGTH: 13 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96: Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Leu Leu INFORMATION FOR SEQ ID NO:97: SEQUENCE CHARACTERISTICS: LENGTH: 14 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97: Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Leu Leu INFORMATION FOR SEQ ID NO:98: SEQUENCE CHARACTERISTICS: LENGTH: 16 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear PCT/US95/00714 SUBSTITUTE SHEET (RULE 26) R WO 95/19370 104 PCT/US95/00714 (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98: Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu Arg Arg INFORMATION FOR EEQ ID NO:99: SEQUENCE CHARACTERISTICS: LENGTH: 16 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99: Lys Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Leu Leu INFORMATION FOR SEQ ID NO:100: SEQUENCE CHARACTERISTICS: LENGTH: 1'5 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Asn INFORMATION FOR SEQ ID NO:101: SEQUENCE CHARACTERISTICS: LENGTH: 15 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear SUBSTITUTE SHEET (RULE 26) WO 95/19370 0 5 CT/US95/00714 (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is homoserine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:101: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Xaa INFORMATION FOR SEQ ID NO:102: SEQUENCE CHARACTERISTICS: LENGTH: 18 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102: Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys Asn Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:103: SEQUENCE CHARACTERISTICS: LENGTH: 18 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103: Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys Pro Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:104: SEQUENCE CHARACTERISTICS: LENGTH: 22 amino acids TYPE: amino acid SUBSTITUTE SHEET (RULE 26) W095/19370 106 PCT/US95/00714

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104: Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu Gin Gly Pro Pro Gin Gly Gin Ser Pro Gin INFORMATION FOR SEQ ID NO:105: SEQUENCE CHARACTERISTICS: LENGTH: 20 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105: Leu Ala Ser Lys Ala Gly Ala Ile Ala Gly Lys Ile Ala Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:106: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106: Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:107: SEQUENCE CHARACTERISTICS: LENGTH: 8 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear SUBSTITUTE SHEET (RULE 26) WO 95/19370 107 PCT/US95/00714 (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107: Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:108: SEQUENCE CHARACTERISTICS: LENGTH: 9 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108: Lys Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:109: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE; amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:109: Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:110: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:111: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids SUBSTITUTE SHEET (RULE 28)

I-~

WO 95/19370 108 TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11l: Ala Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:112: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112: Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:113: SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113: Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:114: SEQUENCE CHARACTERISTICS: LENGTH: 13 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114: PCT/US95/00714 SUBSTITUTE SHEET (RULE 26) t WO 95/19370 109 PCTIUS95/00714 Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:115: SEQUENCE CHARACTERISTICS: LENGTH: 14 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:115: Lys Lys Lea Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:116: SEQUENCE CHARACTERISTICS: LENGTH: 15 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:117: SEQUENCE CHARACTERISTICS: LENGTH: 14 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Arg SUBSTITUTE SHEET (RULE 28) -9 WO 95/19370 110 INFORMATION FOR SEQ ID NO:118: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118: Lys Phe Ala Lys Lys Phe Ala INFORMATION FOR SEQ ID NO:119: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119: Lys Ile Ala Lys Lys Ile Ala INFORMATION FOR SEQ ID NO:120: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: uFptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120: Arg Phe Ala Arg Arg Phe Ala INFORMATION FOR SEQ ID NO:121: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear PCT/US95/00714 SUBSTITUTE SHEET (RULE 28) P ~~DT -I WO 95/19370 111 (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121: Lys Phe Ala Lys Phe Ala Lys Lys Phe Ala INFORMATION FOR SEQ ID NO:122: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122: Lys Lys Phe Ala Lys Phe Ala Lys Lys Phe Ala INFORMATION FOR SEQ ID NO:123: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123: Lys Lys Leu Ala Lys Leu Ala Lys Lys Leu Ala INFORMATION FOR SEQ ID NO:124: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124: Lys Leu Ala Lys Leu Ala Lys Lys Leu Ala INFORMATION FOR SEQ ID NO:125: SEQUENCE CHARACTERISTICS: LENGTH: 14 amino acids PCT/US95/00714 SUBSTITUTE SHEET (RULE 26) c WO 95119370 112 PCT/US95/0071, TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125: Lys Phe Ala Lys Lys Phe Ala Lys Phe Ala Lys Lys Phe Ala INFORMATION FOR SEQ ID NO:126: SEQUENCE CHARACTERISTICS: LENGTH: 14 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126: Arg Phe Ala Arg Arg Phe Ala Arg Phe Ala Arg Arg Phe Ala INFORMATION FOR SEQ ID NO:127: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127: Glu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:128: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128: SUBSTITUTE SHEET (RULE 28) Wd 95/19370 113 Lys Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:129: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129: Lys Leu Lys Lys Lys Phe Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:130: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130: Leu Lys Lys Leu Leu Glu Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:131: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131: Leu Lys Lys Leu Leu Lys Glu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:132: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid PCT/US95/00714 SUBSTITUTE SHEET (RULE 28) wnO'o/ll R7l WrC T/US95/00714 114

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:132: Xaa Xaa Leu Leu Xaa Glu Leu Xaa Xaa Leu INFORMATION FOR SEQ ID NO:133: SEQUENCE CHARACTERISTICS: LENGTH: 12 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Cys INFORMATION FOR SEQ ID NO:134: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:134: Xaa Xaa Leu Leu Xaa Asp Leu Xaa Xaa Leu INFORMATION FOR SEQ ID NO:135: SEQUENCE CHARACTERISTICS: LENGTH: 13 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear SUBSTITUTE SHEET (RULE 28) WO 95/19370 115 PCT/US95/00714 (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135: Lys Lys Phe Gly Lys Lys Phe Val Lys Ile Leu Lys Lys INFORMATION FOR SEQ ID NO:136: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136: Lys Trp Lys Leu Phe Lys Lys Ile Glu Lys Val INFORMATION FOR SEQ ID NO:137: SEQUENCE CHARACTERISTICS: LENGTH: 7 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137: Leu Lys Lys Leu Leu Lys Lys INFORMATION FOR SEQ ID NO:138: SEQUENCE CHARACTERISTICS: LENGTH: 9 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138: Leu Lys Lys Leu Leu Lys Leu Leu Lys INFORMATION FOR SEQ ID NO:139: SEQUENCE CHARACTERISTICS: LENGTH: 14 amino acids SUBSTITUTE SHEET (RULE 28) 0 W 95119370U PCT/US95/00714 116 TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139: Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:140: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:140: Leu Xaa Leu Leu Xaa Xaa Leu Xaa Xaa Leu INFORMATION FOR SEQ ID NO:141: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:141: Leu Xaa Lys Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOi SEQ ID NO:142: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear SUBSTITUTE SHEET (RULE 28) I* n.t inrfln n Wv Y31A3yV 117 PC (ii) MOLECULE TYPE: peptide (ix) FEATURE: OTHER INFORMATION: Xaa is ornithine.

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:142: Leu Xaa Xaa Leu Leu Xaa Xaa Leu Xaa Xaa Leu INFORMATION FOR SEQ ID NO:143: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:143: Xaa Xaa Leu Leu Xaa Xaa Leu Xaa Xaa Leu INFORMATION FOR SEQ ID NO:144: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid

STRANDEDNESS:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:144: Xaa Xaa Leu Leu Xaa Gin Leu Xaa Xaa Leu INFORMATION FOR SEQ ID NO:145: SEQUENCE CHARACTERISTICS: LENGTH: 9 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear TUS95/00714 SUBSTITUTE SHEET (RULE 28) WO 9 5/1o() "o PCT/IUS95/00714 118 (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145: Arg Leu Leu Arg Arg Leu Arg Arg Leu INFORMATION FOR SEQ ID NO:146: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146: Val Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:147: SEQUENCE CHARACTERISTICS: LENGTH: 14 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147: Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu INFORMATION FOR SEQ ID NO:148: SEQUENCE CHARACTERISTICS: LENGTH: 10 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148: Lys Leu Lys Lys Leu Lys Lys Leu Phe Lys INFORMATION FOR SEQ ID NO:149: SEQUENCE CHARACTERISTICS: LENGTH: 16 amino acids SUBSTITUTE SHEET (RULE 28) WO 95/19370 119 PCT/US95/00714 TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:149: Gly Ile Lys Lys Phe Leu Lys Lys Ala Gly Lys Phe Gly Lys Ala Phe 10 INFORMATION FOR SEQ ID NO:150: SEQUENCE CHARACTERISTICS: LENGTH: 20 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150: Ile Ala Gly Ala Ile Ala Lys Ile Ala Gly Lys Ile Ala Lys Ile Ala Gly Ala Ile Ala INFORMATION FOR SEQ ID NO:151: SEQUENCE CHARACTERISTICS: LENGTH: 11 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151: Leu Lys Lys Leu Leu Lys Glu Leu Leu Lys Leu INFORMATION FOR SEQ ID NO:152: SEQUENCE CHARACTERISTICS: LENGTH: 21 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear SUBSTITUTE SHEET (RULE 28) WO 95/19370 120 (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152: Lys Val Ala Leu Lys Ala Leu Lys Lys Val Ala Leu Lys Ala Leu Lys Val Ala Leu Lys Ala Leu INFORMATION FOR SEQ ID NO:153: SEQUENCE CHARACTERISTICS: LENGTH: 14 amino acids TYPE: amino acid

STRANDEDNESS:

TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153: Lys Ile Ala Lys Lys Ile Ala Lys Ile Ala Lys Lys Ile Ala PCTIUS9S/00714 SUBSTITUTE SHEET (RULE 26)

Claims

1. formula: An N-terminal substituted peptide having the H T-N-X wherein X is a biologically active peptide having the following structure: (SEQ ID NO: 143); T is a lipophilic moiety, wherein if T is 0 1 IS S *c r *o 55 5555* I. 5 *s *e S S S *c S o S I R is hydrocarbon having at least 2 carbon atoms.

2. A peptide according to claim 1, wherein the N- terminal substituted peptide is N-terminal substituted with octanoyl, and X is a biologically active peptide having the following structure: (SEQ ID NO: 143).

3. A composition for inhibiting growth of a target cell, virus, or virally-infected cell, comprising: a N-terminal substituted peptide according to claim 1 or claim 2; and an acceptable pharmaceutical carrier, wherein the N-terminal substituted peptide is present in an ar..unt effective to inhibit growth of the target cell, H;\Luia\Keep\rp*ci\1728,.95.ACAININdoc 14/05/98 s, 122 virus, or virally-infected cell.

4. An endotoxin-neutralising composition comprising a peptide according to claim 1 or claim 2, together with a pharmaceutically-acceptable carrier. A method of inhibiting growth of a target cell, virus, or virally-infected cell in a host, comprising the step of administering a growth-inhibitory amount of a peptide according to claim 1 or claim 2, to a subject in need of such treatment.

6. A method of treatment of septic shock, comprising the step of administering an effective amount of a peptide according to claim 1 or claim 2, to a subject in need of such treatment.

7. A method of neutralising endotoxin, comprising the step of administering an effective amount of a peptide 20 according to claim 1 or claim 2, to a subject suffering adverse symptoms caused endotoxin.

8. A method of inhibiting growth of a target cell, virus, or virally-infected cell in a host, comprising the 25 step of administering a growth-inhibitory amount of a composition according to claim 3 or claim 4, to a subject o: in need of such treatment.

9. A method of treatment of septic shock, comprising the step of administering an effective amount of a composition according to claim 3 or claim 4, to a subject in need of such treatment. A method of neutralising endotoxin, comprising the step of administering an effective amount of a composition according to claim 3 or claim 4, to a subject suffering adverse symptoms caused endotoxin. Hi\Luisa\Keep\spacia\17288.95.MAGANIN.toc 14/05/98 IA^ 6- 0P i cc- 123

11. A peptide according to claim 1, substantially as hereinbefore described with reference to any one of the examples. Dated this 13th day of May 1.998 MAGAININ PHARMACEUTICALS INC. By Its Patent Attorneys: GRIFFITH HACK Fellows Institute of Patent Attorneys of Australia 0* 4 4 4. 4 4 4*4 44@*44 4 4

444. 4. @4 4 4 1~ '4' Hn.\LUiSa\KeeP\SPeCiS\i:*V dS.MAGAININ.dc 14/05/98l