CA2208420C - Methods and compositions for inhibition of membrane fusion-associated events, including hiv transmission - Google Patents

Abstract

The present invention relates to peptides which exhibit potent anti-retroviral activity. The peptides of the invention comprise DP178 (SEQ ID:1) peptide corresponding to amino acids 638 to 673 of the HIV-1LAI
gp4l protein, and fragments, analogs and homologs of DP178.
The invention further relates to the uses of such peptides as inhibitory of human and non-human retroviral, especially HIV, transmission to uninfected cells.

Description

DEMANDES OU BREVETS VOLUMINEUX

LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET
COMPREND PLUS D`UN TOME.

NOTE: Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets ZZGSS9/ZC~
JUMBO APPLICATIONS/PATENTS

THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE
THAN ONE VOLUME

THIS IS VOLUME OF 2"

NOTE: For additional volumes-please contact-thd Canadian Patent Office METHODS AND COMPOSITIONS FOR INHIBITION OF 'NEVDMME
FUSION^ASSOC~ATED EVENTS YNCLUD r YTIV TRANSMISSION

L_ INTRODUCTION
3.5 The present invention relates, first, to DP178 (SEQ ID No ;1) , a peptide corresponding to amino acids 638 to 673 of the HIV-1x,,11 transmembrane protein (TM) gp41, and portions or analogs of DP178 (SEQ ID NO:1), which exhibit anti-membrane fusion capability, antiviral activity, such as the ability to inhibit HIV
transmission to uninfected CD-4't cells, or an ability to modulate intracellular processes involving coiled-coil peptide structures. Further, the invention relates to the use of DPI78 (SEQ ID N0:1) and DP178 portions and/or analogs as antifusogenic or antiviral compounds or as inhibitors of intracellular events involving coiled-coil peptide structures. The present invention also relates to peptides analogous to DPI07 (SEQ ID H0;25), a peptide corresponding to amino acids 558 to 595 of the HIV-11A, transmembrane protein - (TM) gp4l, having amino acid sequences present in other viruses, such as enveloped viruses; and/or other organisms, and further relates to the uses of such peptides. These peptides exhibit anti membrane fusion capability, antiviral activity-, or the ability to modulate intracellular processes involving coiled-coil peptide structures. The present invention additionally relates to methods for identifying compounds that disrupt the interaction between DP178 and DP107, and/or between DP107-like and DP178-like s peptides. Further, the invention relates to the use of the peptides of the invention as diagnostic agents.
For example, a DP178 peptide may be used as an HIV
subtype-specific diagnostic. The invention is demonstrated, first, by way of an Example wherein DP178 (SEQ ID:1), and a peptide whose sequence is homologous to DP178 are each shown to be potent, non-cytotoxic inhibitors of HIV-1 transfer to uninfected CD-4+ cells. The invention is further demonstrated by Examples wherein peptides having structural and/or is amino acid motif similarity to DP107 and DP178 are identified in a variety of viral and nonviral organisms, and in examples wherein a number of such identified peptides derived from several different viral systems are demonstrated to exhibit antiviral activity.

2. BACKGROUND OF THE INVENTION
2.1 MEMBRANE FUSION EVENTS
Membrane fusion is a ubiquitous cell biological process (for a review, see White, J.M., 1992, Science 258:917-924). Fusion events which mediate cellular housekeeping functions, such as endocytosis, constitutive secretion, and recycling of membrane components, occur continuously in all eukaryotic cells.
Additional fusion events occur in specialized cells. Intracellularly, for example, fusion events are involved in such processes as occur in regulated exocytosis of hormones, enzymes and neurotransmitters.

WO 96/19495 _ - PCTIUS95/16733 Intercellularly, such fusion events feature prominently in, for example, sperm-egg fusion and myoblast fusion.
= Fusion events are also associated with disease states. For example, fusion events are involved in the formation of giant cells during inflammatory reactions, the entry of all enveloped viruses into cells, and, in the case of human immunodeficiency virus (HIV), for example, are responsible for the virally induced cell-cell fusion which leads to cell death.

2.2. THE HUMAN IMMUNODEFICIENCY VIRUS
The human immunodeficiency virus (HIV) has been implicated as the primary cause of the slowly degenerative immune system disease termed acquired immune deficiency syndrome (AIDS) (Barre-Sinoussi, F.
et al., 1983, Science 220:868-870; Gallo, R. et al., 1984, Science 224:500-503). There are at least two distinct types of HIV: HIV-1 (Barre-Sinoussi, F. et al., 1983, Science 220:868-870; Gallo R. et al., 1984, Science 224:500-503) and HIV-2 (Clavel, F. et al., 1986, Science 233:343-346; Guyader, M. et al., 1987, Nature 326:662-669). Further, a large amount of genetic heterogeneity exists within populations of each of these types. Infection of human CD-4+ T-lymphocytes with an HIV virus leads to depletion of the cell type and eventually to opportunistic infections, neurological dysfunctions, neoplastic growth, and ultimately death.
HIV is a member of the lentivirus family of retroviruses (Teich, N. et al., 1984, RNA Tumor Viruses, Weiss, R. et al., eds., CSH-Press, pp. 949-956). Retroviruses are small enveloped viruses that contain a diploid, single-stranded RNA genome, and replicate via a DNA intermediate produced by a virally-encoded reverse transcriptase, an RNA-dependent DNA polymerase (Varmus, H., 1988, Science 240:1427-1439). Other retroviruses include, for example, oncogenic viruses such as human T-cell leukemia viruses (HTLV-I,-II,-III), and feline leukemia virus.
The HIV viral particle consists of a viral core, composed of capsid proteins, that contains the viral RNA genome and those enzymes required for early replicative events. Myristylated Gag protein forms an outer viral shell around the viral core, which is, in turn, surrounded by a lipid membrane enveloped derived from the infected cell membrane. The HIV enveloped surface glycoproteins are synthesized as a single 160 Kd precursor protein which is cleaved by a cellular protease during viral budding into two glycoproteins, gp4l and gp120. gp4l is a transmembrane protein and gp120 is an extracellular protein which remains non-covalently associated with gp4l, possibly in a trimeric or multimeric form (Hammarskjold, M. and Rekosh, D., 1989, Biochem. Biophys. Acta 989:269-280).
HIV is targeted to CD-4+ cells because the CD-4 cell surface protein acts as the cellular receptor for the HIV-1 virus (Dalgleish, A. et al., 1984, Nature 312:763-767; Klatzmann et al., 1984, Nature 312:767-768; Maddon et al., 1986, Cell 47:333-348). Viral entry into cells is dependent upon gp120 binding the cellular CD-4+ receptor molecules (McDougal, J.S. et _, 1986, Science 231:382-385; Maddon, P.J. et al., 1986, Cell 47:333-348) and thus explains HIV's tropism for CD-4+ cells, while gp4l anchors the enveloped glycoprotein complex in the viral membrane.

2.3. HIV TREATMENT
HIV infection is pandemic and HIV associated diseases represent a major world health problem.
Although considerable effort is being put into the successful design of effective therapeutics, currently no curative anti-retroviral drugs against AIDS exist.
In attempts to develop such drugs, several stages of the HIV life cycle have been considered as targets for therapeutic intervention (Mitsuya, H. et al., 1991, FASEB J. 5:2369-2381). For example, virally encoded reverse transcriptase has been one focus of drug development. A number of reverse-transcriptase-targeted drugs, including 2',3'-dideoxynucleoside analogs such as AZT, ddl, ddC, and d4T have been developed which have been shown to been active against HIV (Mitsuya, H. et al., 1991, Science 249:1533-1544).
While beneficial, these nucleoside analogs are not curative, probably due to the rapid appearance of drug resistant HIV mutants (Lander, B. et al., 1989, Science 243:1731-1734). In addition, the drugs often exhibit toxic side effects such as bone marrow suppression, vomiting, and liver function abnormalities.
Attempts are also being made to develop drugs which can inhibit viral entry into the cell, the earliest stage of HIV infection. Here, the focus has thus far been on CD4, the cell surface receptor for HIV. Recombinant soluble CD4, for example, has.been shown to inhibit infection of CD-4+ T-cells by some' HIV-1 strains (Smith, D.H. et al., 1987, science 238:1704-1707). Certain primary HIV-1 isolates, however, are relatively less sensitive to inhibition by recombinant CD-4 (Daar, E. et al., 1990, Proc.
Natl. Acad. Sci. USA 87:6574-6579). In addition, recombinant soluble CD-4 clinical trials have produced inconclusive results (Schooley, R. et al., 1990, Ann.
Int. Med. 112:247-253; Kahn, J.O. et al., 1990, Ann.
Int. Med. 112:254-261; Yarchoan, R. et al., 1989, Proc. Vth Int. Conf. on AIDS, p. 564, MCP 137).
The late stages of HIV replication, which involve crucial virus-specific secondary processing of certain viral proteins, have also been suggested as possible anti-HIV drug targets. Late stage processing is dependent on the activity of a viral protease, and drugs are being developed which inhibit this protease (Erickson, J., 1990, Science 249:527-533). The clinical outcome of these candidate drugs is still in question.
Attention is also being given to the development of vaccines for the treatment of HIV infection. The HIV-1 enveloped proteins (gp160, gp120, gp4l) have been shown to be the major antigens for anti-HIV
antibodies present in AIDS patients (Barin, et al., 1985, Science 228:1094-1096). Thus far, therefore, these proteins seem to be the most promising candidates to act as antigens for anti-HIV vaccine development. To this end, several groups have begun to use various portions of gp160, gp120, and/or gp4l as immunogenic targets for the host immune system.
See for example, Ivanoff, L. et al., U.S. Pat. No.
5,141,867; Saith, G. et al., WO 92/22,654; Shafferman, A., WO 91/09,872; Formoso, C. et al., WO 90/07,119.
Clinical results concerning these candidate vaccines, however, still remain far in the future.
Thus, although a great deal of effort is being directed to the design and testing of anti-retroviral drugs, a truly effective, non-toxic treatment is still needed.

3. SUMMARY OF THE INVENTION
The present invention relates, first, to DP178 (SEQ ID:1), a 36-amino acid synthetic peptide corresponding to amino acids 638 to 673 of the transmembrane protein (TM) gp4l from the HIV-1 isolate LAI (HIV-lUI), which exhibits potent anti-HIV-1 activity. As evidenced by the Example presented below, in Section 6, the DP178 (SEQ ID:1) antiviral activity is so high that, on a weight basis, no other known anti-HIV agent is effective at concentrations as low as those at which DP178 (SEQ ID:1) exhibits its inhibitory effects.
The invention further relates to those portions and analogs of DP178 which also show such antiviral activity, and/or show anti-membrane fusion capability, or an ability to modulate intracellular processes involving coiled-coil peptide structures. The term "DP178 analog" refers to a peptide which contains an amino acid sequence corresponding to the DP178 peptide sequence present within the gp4l protein of HIV-llp,1, but found in viruses and/or organisms other than HIV-lI.AI. Such DP178 analog peptides may, therefore, correspond to DP178-like amino acid sequences present in other viruses, such as, for example, enveloped viruses, such as retroviruses other than HIV-lIAI, as well as non-enveloped viruses. Further, such analogous DP178 peptides may also correspond to DP178-like amino acid sequences present in nonviral organisms.
The invention further relates to peptides DP107 (SEQ ID NO:25) analogs. DP107 is a peptide corresponding to amino acids 558-595 of the HIV-lIA1 transmembrane protein (TM) gp4l. The term "DP107 analog" as used herein refers to a peptide which contains an amino acid sequence corresponding to the DP107 peptide sequence present within the gp4l protein of HIV-l,1, but found in viruses and organisms other than HIV-lLA1. Such DP107 analog peptides may, therefore, correspond to DP107-like amino acid sequences present in other viruses, such as, for for example, enveloped viruses, such as retroviruses other than HIV-lLA1, as well as non-enveloped viruses.
Further, such DP107 analog peptides may also correspond to DP107-like amino acid sequences present in nonviral organisms.
Further, the peptides of the invention include DP107 analog and DP178 analog peptides having amino acid sequences recognized or identified by the 107x178x4, ALLMOTI5 and/or PLZIP search motifs described herein.
The peptides of the invention may, for example, exhibit antifusogenic activity, antiviral activity, and/or may have the ability to modulate intracellular processes which involve coiled-coil peptide structures. With respect to the antiviral activity of the peptides of the invention, such an antiviral activity includes, but is not limited to the inhibition of HIV transmission to uninfected CD-4+
cells. Additionally, the antifusogenic capability, antiviral activity or intracellular modulatory activity of the peptides of the invention merely requires the presence of the peptides of the invention, and, specifically, does not require the stimulation of a host immune response directed against such peptides.
The peptides of the invention may be used, for example, as inhibitors of membrane fusion-asociated events, such as, for example, the inhibition of human and non-human retroviral, especially HIV, transmission to uninfected cells. It is further contemplated that the peptides of the invention may be used as modulators of intracellular events involving coiled-coil peptide structures.
The peptides of the invention may, alternatively, be used to identify compounds which may themselves exhibit antifusogenic, antiviral, or intracellular modulatory activity. Additional uses include, for example, the use of the peptides of the invention as organism or viral type and/or subtype-specific diagnostic tools.
The terms "antifusogenic" and "anti-membrane fusion", as used herein, refer to an agent's ability to inhibit or reduce the level of membrane fusion events between two or more moieties relative to the level of membrane fusion which occurs between said moieties in the absence of the peptide. The moieties may be, for example, cell membranes or viral structures, such as viral envelopes or pili. The term "antiviral", as used herein, refers to the compound's ability to inhibit viral infection of cells, via, for example, cell-cell fusion or free virus infection.
Such infection may involve membrane fusion, as occurs in the case of enveloped viruses, or some other fusion event involving a viral structure and a cellular structure (e.g., such as the fusion of a viral pilus and bacterial membrane during bacterial conjugation).
It is also contemplated that the peptides of the invention may exhibit the ability to modulate intracellular events involving coiled-coil peptide structures. "Modulate", as used herein, refers to a stimulatory or inhibitory effect on the intracellular process of interest relative to the level or activity of such a process in the absence of a peptide of the invention.

Embodiments of the invention are demonstrated below wherein an extremely low concentration of DP178 (SEQ ID:1), and very low concentrations of a DP178 homolog (SEQ ID:3) are shown to be potent inhibitors of HIV-1 mediated CD-4+ cell-cell fusion (i.e., syncytial formation) and infection of CD-4+ cells by cell-free virus. Further, it is shown that DP178 (SEQ
ID:1) is not toxic to cells, even at concentrations 3 logs higher than the inhibitory DP-178 (SEQ ID:1) concentration. %
The present invention is based, in part, on the surprising discovery that the DP107 and DP178 domains of the HIV gp4l protein non-covalently complex with each other, and that their interaction is required for the normal infectivity of the virus. This discovery is described in the Example presented, below, in Section 8. The invention, therefore, further relates to methods for identifying antifusogenic, including antiviral, compounds that disrupt the interaction between DP107 and DP178, and/or between DP107-like and DP178-like peptides.
Additional embodiments of the invention (specifically, the Examples presents in Sections 9-16 and 19-25, below) are demonstrated, below, wherein peptides, from a variety of viral and nonviral sources, having structural and/or amino acid motif similarity to DP107 and DP178 are identified, and search motifs for their identification are described.
Further, Examples (in Sections 17, 18, 25-29) are presented wherein.a number of the peptides of the invention are demonstrated exhibit substantial antiviral activity or activity predictive of antiviral activity. =

3.1. DEFINITIONS
Peptides are defined herein as organic compounds comprising two or more amino acids covalently joined by peptide bonds. Peptides may be referred to with respect to the number of constituent amino acids, i.e., a dipeptide contains two amino acid residues, a tripeptide contains three, etc. Peptides containing ten or fewer amino acids may be referred to as oligopeptides, while those with more than ten amino acid residues are polypeptides. Such peptides may also include any of the modifications and additional amino and carboxy groups as are described herein.
Peptide sequences defined herein are represented by one-letter symbols for amino acid residues as follows:
A (alanine) R (arginine) N (asparagine) D (aspartic acid) C (cysteine) Q (glutamine) E (glutamic acid) G (glycine) H (histidine) I (isoleucine) L (leucine) K (lysine) M (methionine) F (phenylalanine) P (proline) S (serine) T (threonine) W (tryptophan) Y (tyrosine) V (valine) 4. BRIEF DESCRIPTION OF THE FIGURES
FIG. 1. Amino acid sequence of DP178 (SEQ ID:1) derived from HIVI1,,I; DP178 homologs derived from HIV-1SF2 (DP-185; SEQ ID:3), HIV-1RF (SEQ ID:4), and HIV-1mN
(SEQ ID:5); DP178 homologs derived from amino acid sequences of two prototypic HIV-2 isolates, namely, HIV-2wd (SEQ ID: 6) and HIV-21,I, (SEQ ID: 7) ; control peptides: DP-180 (SEQ ID:2), a peptide incorporating the amino acid residues of DP178 in a scrambled sequence; DP-118 (SEQ ID:10) unrelated to DP178, which inhibits HIV-1 cell free virus infection; DP-125 (SEQ
ID:8), unrelated to DP178, also inhibits HIV-1 cell free virus infection; DP-116 (SEQ ID:9), unrelated to DP178, is negative for inhibition of HIV-1 infection when tested using a cell-free virus infection assay.
Throughout the figures, the one letter amino acid code is used.
FIG. 2. Inhibition of HIV-1 cell-free virus infection by synthetic peptides. IC50 refers to the concentration of peptide that inhibits RT production from infected cells by 50% compared to the untreated control. Control: the level of RT produced by untreated cell cultures infected with the same level of virus as treated cultures.
FIG. 3. Inhibition of HIV-1 and HIV-2 cell-free virus infection by the synthetic peptide DP178 (SEQ
ID:1). IC50: concentration of peptide that inhibits RT production by 50% compared to the untreated control. Control: Level of RT produced by untreated cell cultures infected with the same level of virus as treated cultures.
FIG. 4A-4B. Fusion Inhibition Assays. FIG 4A:
DP178 (SEQ ID:1) inhibition of HIV-1 prototypic isolate-mediated syncytial formation; data represents the number of virus-induced syncytial per cell. FIG.

4B: DP-180 (SEQ ID:2) represents a scrambled control peptide; DP-185 (SEQ ID:3) represents a DP178 homolog derived from HIV-lsF2 isolate; Control, refers to the number of syncytial produced in the absence of peptide.
FIG. 5. Fusion inhibition assay: HIV-1 vs.
HIV-2. Data represents the number of virus-induced syncytial per well. ND: not done.
FIG. 6. Cytotoxicity study of DP178 (SEQ ID:1) and DP-116 (SEQ ID:9) on CEM cells. Cell proliferation data is shown.
FIG. 7. Schematic representation of HIV-gp4l and maltose binding protein (MBP)-gp4l fusion proteins. DP107 and DP178 are synthetic peptides based on the two putative helices of gp4l. The letter P in the DP107 boxes denotes an Ile to Pro mutation at amino acid number 578. Amino acid residues are numbered according to Meyers et al., "Human Retroviruses and AIDS", 1991, Theoret. Biol. and Biophys. Group, Los Alamos Natl. Lab., Los Alamos, NM.
The proteins are more fully described, below, in Section 8.1.1.
FIG. 8. A point mutation alters the conformation and anti-HIV activity of M41.
FIG. 9. Abrogation of DP178 anti-HIV activity.
Cell fusion assays were carried out in the presence of 10 nM DP178 and various concentrations of M41O178 or M41PO178.
FIG. 10. Binding of DP178 to leucine zipper of gp4l analyzed by FAb-D ELISA.
FIG. 11A-B. Models for a structural transition in the HIV-1 TM protein. Two models are proposed which indicate a structural transition from a native oligomer to a fusogenic state following a trigger event (possibly gpl20 binding to CD4). Common features of both models include (1) the native state is held together by noncovalent protein-protein interactions to form the heterodimer of gp120/41 and other interactions, principally though gp41 interactive sites, to form homo-oligomers on the virus surface of the gp120/41 complexes; (2) shielding of the hydrophobic fusogenic peptide at the N-terminus (F) in the native state; and (3) the leucine zipper domain (DP107) exists as a homo-oligomer coiled coil only in the fusogenic state. The major differences in the two models include the structural state (native or fusogenic) in which the DP107 and DP178 domains are complexed to each other. In the first model (FIG.
11A) this interaction occurs in the native state and in the second (FIG. 11B), it occurs during the fusogenic state. When triggered, the fusion complex in the model depicted in (A) is generated through formation of coiled-coil interactions in homologous DP107 domains resulting in an extended a-helix. This conformational change positions the fusion peptide for interaction with the cell membrane. In the second model (FIG. 11B), the fusogenic complex is stabilized by the association of the DP178 domain with the DP107 coiled-coil.
FIG. 12. Motif design using heptad repeat positioning of amino acids of known coiled-coils.
FIG. 13. Motif design using proposed heptad repeat positioning of amino acids of DP107 and DP178.
FIG. 14. Hybrid motif design crossing GCN4 and DP107.
FIG. 15. Hybrid motif design crossing GCN4 and DP178.
FIG. 16. Hybrid motif design 107x178x4, crossing DP107 and DP178. This motif was found to be the most consistent at identifying relevant DP107-like and DP178-like peptide regions.
FIG. 17. Hybrid motif design crossing GCN4, DP107, and DP178.
FIG. 18. Hybrid motif design ALLMOTI5 crossing GCN4, DP107, DP178, c-Fos c-Jun, c-Myc, and Flu Loop 36.
FIG. 19. PLZIP motifs designed to identify N-terminal proline-leucine zipper motifs.
FIG. 20. Search results for HIV-1 (BRU
isolate) enveloped protein gp4l. Sequence search motif designations: Spades (4): 107x178x4; Hearts (V) ALLMOTI5; Clubs (4): PLZIP; Diamonds (+):
transmembrane region (the putative transmembrane domains were identified using a PC/Gene program designed to search for such peptide regions).
Asterisk (*): Lupas method. The amino acid sequences identified by each motif are bracketed by the respective characters. Representative sequences chosen based on 107x178x4 searches are underlined and in bold. DP107 and DP178 sequences are marked, and additionally double-underlined and italicized.
FIG. 21. Search results for human respiratory syncytial virus (RSV) strain A2 fusion glycoprotein Fl. Sequence search motif designations are as in FIG. 20.
FIG. 22. Search results for simian immunodeficiency virus (SIV) enveloped protein gp4l (AGM3 isolate). Sequence search motif designations are as in FIG. 20.
FIG. 23. Search results for canine distemper virus (strain Onderstepoort) fusion glycoprotein 1. Sequence search motif designations are as in FIG. 20.

FIG. 24. Search results for newcastle disease virus (strain Australia-Victoria/32) fusion glycoprotein Fl. Sequence search motif designations are as in FIG. 20.
FIG. 25. Search results for human parainfluenza 3 virus (strain NIH 47885) fusion glycoprotein Fl. Sequence search motif designations are as in FIG. 20.
FIG. 26. Search results for influenza A
virus (strain A/AICHI/2/68) hemagglutinin precursor HA2. Sequence search designations are as in FIG. 20.
FIG. 27A-F. Respiratory Syncytial Virus (RSV) peptide antiviral and circular dichroism data.
FIG. 27A-C: Peptides derived from the F2 DP178/DP107-like region. Antiviral and CD data. FIG. 27D-F:
Peptides derived from the F1 DP107-like region.
Peptide and CD data.
Antiviral activity (AV) is represented by the following qualitative symbols:
"-", negative antiviral activity;
"+/-tI, antiviral activity at greater than 100 g/ml;
antiviral activity at between 50-100 g/ml;
"++", antiviral activity at between 20-50 g/ml;
"+++", antiviral activity at between 1-20 g/ml;
antiviral activity at <l g/ml.
CD data, referring to the level of helicity is represented by the following qualitative symbol:
"-" no helicity;
"+", 25-50% helicity;
It++", 50-75% helicity;
"+++"' 75-100% helicity.
IC50 refers to the concentration of peptide necessary to produce only 50% of the number of syncytial relative to infected control cultures RECTIFIED SHEET (RULE 91) containing no peptide. IC50 values were obtained using purified peptides only.
FIG. 28A-C. Respiratory Syncytial Virus (RSV) DP178-like region (F1) peptide antiviral and CD
data. Antiviral symbols, CD symbols, and IC50 are as in FIG. 27A-F. IC50 values were obtained using purified peptides only.
FIG. 29A-E. Peptides derived from the HPIV3 F1 DP107-like region. Peptide antiviral and CD data.
Antiviral symbols, CD symbols, and IC50 are as in FIG.
27A-F. Purified peptides were used to obtain IC50 values, except where the values are marked by an asterisk (*), in which cases, the IC50 values were obtained using a crude peptide preparation.
FIG. 30A-C. Peptides derived from the HPIV3 F1 DP178-like region. Peptide antiviral and CD data.
Antiviral symbols, CD symbols, and IC50 are as in FIG.
27A-F. Purified peptides were used to obtain IC50 values, except where the values are marked by an asterisk (*), in which cases, the IC50 values were obtained using a crude peptide preparation.
FIG. 31. Motif search results for simian immunodeficiency virus (SIV) isolate MM251, enveloped polyprotein gp4l. Sequence search designations are as in FIG. 20.
FIG. 32. Motif search results for Epstein-Barr Virus (Strain B95-8), glycoprotein gp110 precursor (designated gp115). BALF4. Sequence search designations are as in FIG. 20.
FIG. 33. Motif search results for Epstein-Barr Virus (Strain B95-8), BZLF1 trans-activator protein (designated EB1 or Zebra). Sequence search designations are as in FIG. 20. Additionally, "@"
refers to a well known DNA binding domain and 11+11 refers to a well known dimerization domain, as defined RECTIFIED SHEET (RULE 91) by Flemington and Speck (Flemington, E. and Speck, S.H., 1990, Proc. Natl. Acad. Sci. USA 87:9459-9463).
FIG. 34. Motif search results for measles virus (strain Edmonston), fusion glycoprotein Fl.
Sequence search designations are as in FIG. 20.
FIG. 35. Motif search results for Hepatitis B Virus (Subtype AYW), major surface antigen precursor S. Sequence search designations are as in FIG. 20.
FIG. 36. Motif search results for simian Mason-Pfizer monkey virus, enveloped (TM) protein gp20. Sequence search designations are as in FIG. 20.
FIG. 37. Motif search results for Pseudomonas aerginosa, fimbrial protein (Pilin).
Sequence search designations are as in FIG. 20.
FIG. 38. Motif search results for Neisseria gonorrhoeae fimbrial protein (Pilin). Sequence search designations are as in FIG. 20.
FIG. 39. Motif search results for Hemophilus influenzae fimbrial protein. Sequence search designations are as in FIG. 20.
FIG. 40. Motif search results for Staphylococcus aureus, toxic shock syndrome toxin-1.
Sequence search designations are as in FIG. 20.
FIG. 41. Motif search results for Staphylococcus aureus enterotoxin Type E. Sequence search designations are as in FIG. 20.
FIG. 42. Motif search results for Staphylococcus aureus enterotoxin A. Sequence search designations are as in FIG. 20.
FIG. 43. Motif search results for Escherichia coli, heat labile enterotoxin A. Sequence search designations are as in FIG. 20.
FIG. 44. Motif search results for human c-fos proto-oncoprotein. Sequence search designations are as in FIG. 20.

FIG. 45. Motif search results for human lupus KU autoantigen protein P70. Sequence search designations are as in FIG. 20.
FIG. 46. Motif search results for human zinc finger protein 10. Sequence search designations are as in FIG. 20.
FIG. 47. Measles virus (MeV) fusion protein DP178-like region antiviral and CD data. Antiviral symbols, CD symbols, and IC50 are as in FIG. 27A-D.
IC50 values were obtained using purified peptides.
FIG. 48. Simian immunodeficiency virus (SIV) TM (fusion) protein DP178-like region antiviral data. Antiviral symbols are as in FIG. 27A-D "NT", not tested.
FIG. 49A-C. DP178-derived peptide antiviral data. The peptides listed herein were derived from the region surrounding the HIV-1 BRU isolate DP178 region (e.g., gp4l amino acid residues 615-717).
In instances where peptides contained DP178 point mutations, the mutated amino acid residues are shown with a shaded background. In instances in which the test peptide has had an amino and/or carboxy-terminal group added or removed (apart from the standard amido-and acetyl- blocking groups found on such peptides), such modifications are indicated. FIG. 49A: The column to the immediate right of the name of the test peptide indicates the size of the test peptide and points out whether the peptide is derived from a one amino acid peptide "walk" across the DP178 region.
The next column to the right indicates whether the test peptide contains a point mutation, while the column to its right indicates whether certain amino acid residues have been added to or removed from the DP178-derived amino acid sequence. FIG 49B: The column to the immediate right of the test peptide name indicates whether the peptide represents a DP178 truncation, the next column to the right points out whether the peptide contains a point mutation, and the column to its right indicates whether the peptide contains amino acids which have been added to or removed from the DP178 sequence itself. FIG. 49C:
The column to the immediate right of the test peptide name indicates whether the test peptide contains a point mutation, while the column to its right indicates whether amino acid residues have been added to or removed from the DP178 sequence itself. IC50 is as defined in FIG. 27A-D, and IC50 values were obtained using purified peptides except where marked with an asterisk (*), in which case the IC50 was obtained using a crude peptide preparation.
FIG. 50. DP107 and DP107 gp4l region truncated peptide antiviral data. IC50 as defined in FIG. 27A-D, and IC50 values were obtained using purified peptides except where marked with an asterisk (*), in which case the IC50 was obtained using a crude peptide preparation.
FIG. 51A-B. Epstein-Barr virus Strain B95-8 BZLF1 DP178/DP107 analog region. peptide walks and electrophoretic mobility shift assay results. The peptides (T-423 to T-446, FIG. 51A; T-447 to T-461, FIG. 51B) represent one amino acid residue "walks"
through the EBV Zebra protein region from amino acid residue 173 to 246.
The amino acid residue within this region which corresponds to the first amino acid residue of each peptide is listed to the left of each peptide, while the amino acid residue within this region which corresponds to the last amino acid residue of each peptide is listed to the right of each peptide. The length of each test peptide is listed at the far right of each line, under the heading "Res".
"ACT" refers to a test peptide's ability to inhibit Zebra binding to its response element. "+"
refers to a visible, but incomplete, abrogation of the response element/Zebra homodimer complex; "+++" refers to a complete abrogation of the complex; and "-"
represents a lack of complex disruption.
FIG. 52A-B. Hepatitis B virus subtype AYW major surface antigen precursor S protein DP178/DP107 analog region and peptide walks. 52A depicts Domain I (S
protein amino acid residues 174-220), which contains a potential DP178/DP107 analog region. In addition, peptides are listed which represent one amino acid peptide "walks" through domain I. 52B depicts Domain II (S protein amino acid residues 233-291), which contains a second potential DP178/DP107 analog region.
In addition, peptides are listed which represent one amino acid peptide "walks" through domain II.

5. DETAILED DESCRIPTION OF THE INVENTION
Described herein are peptides which may exhibit antifusogenic activity, antiviral capability, and/or the ability to modulate intracellular processes involving coiled-coil peptide structures. The peptides described include, first, DP178 (SEQ ID
NO:1), a gp41-derived 36 amino acid peptide and fragments and analogs of DP178.
In addition, the peptides of the invention described herein include peptides which are DP107 analogs. DP107 (SEQ ID NO:25) is a 38 amino acid peptide corresponding to residues 558 to 595 of the HIV-11AI transmembrane (TM) gp4l protein. Such DP107 analogs may exhibit antifusogenic capability, antiviral activity or an ability to modulate intracellular processes involving coiled-coil structures.
Further, peptides of the invention include DP107 and DP178 are described herein having amino acid sequences recognized by the 107x178x4, ALLMOTI5, and PLZIP search motifs. Such motifs are also discussed.
Also described here are antifusogenic, antiviral, intracellular modulatory, and diagnostic uses of the peptides of the invention. Further, procedures are described for the use of the peptides of the invention for the identification of compounds exhibiting antifusogenic, antiviral or intracellular modulatory activity.
While not limited to any theory of operation, the following model is proposed to explain the potent anti-HIV activity of DP178, based, in part, on the experiments described in the Examples, infra. In the HIV protein, gp4l, DP178 corresponds to a putative a-helix region located in the C-terminal end of the gp4l ectodomain, and appears to associate with a distal site on gp4l whose interactive structure is influenced by the leucine zipper motif, a coiled-coil structure, referred to as DP107. The association of these two domains may reflect a molecular linkage or "molecular clasp" intimately involved in the fusion process. It is of interest that mutations in the C-terminal a-helix motif of gp4l (i.e., the D178 domain) tend to enhance the fusion ability of gp4l, whereas mutations in the leucine zipper region (i.e., the DP107 domain) decrease or abolish the fusion ability of the viral protein. It may be that the leucine zipper motif is involved in membrane fusion while the C-terminal a-helix motif serves as a molecular safety to regulate the availability of the leucine zipper during virus-induced membrane fusion.

On the basis of the foregoing, two models are proposed of gp41-mediated membrane fusion which are schematically shown in FIG. 11A-B. The reason for proposing two models is that the temporal nature of the interaction between the regions defined by DP107 and DP178 cannot, as yet, be pinpointed. Each model envisions two conformations for gp4l - one in a "native" state as it might be found on a resting virion. The other in a "fusogenic" state to reflect conformational changes triggered following binding of 120 to CD4 and just gp prior to fusion with the target cell membrane. The strong binding affinity between gp120 and CD4 may actually represent the trigger for the fusion process obviating the need for a pH change such as occurs for viruses that fuse within intracellular vesicles. The two major features of both models are: (1) the leucine zipper sequences (DP107) in each chain of oligomeric enveloped are held apart in the native state and are only allowed access to one another in the fusogenic state so as to form the extremely stable coiled-coils, and (2) association of the DP178 and DP107 sites as they exist in gp4l occur either in the native or fusogenic state. FIG.
11A depicts DP178/DP107 interaction in the native state as a molecular clasp. On the other hand, if one assumes that the most stable form of the enveloped occurs in the fusogenic state, the model in FIG. 11B
can be considered.
When synthesized as peptides, both DP107 and DP178 are potent inhibitors of HIV infection and fusion, probably by virtue of their ability to form complexes with viral gp4l and interfere with its fusogenic process; e.g., during the structural transition of the viral protein from the native structure to the fusogenic state, the DP178 and DP107 peptides may gain access to their respective binding sites on the viral gp4l, aud exert a disruptive influence.

As shown in the Examples, infra, a truncated recombinant gp4l protein corresponding to the ectodomain of gp41 containing both DP107 and DP178 domains (excluding the fusion peptide, transmembrane region and cytoplasmic domain of gp41) did not inhibit HIV-1 induced fusion. However, when a single mutation was introduced to disrupt the coiled-coil structure of the DP107 domain -- a mutation which results in a total loss of biological activity of DP107 peptides --the inactive recombinant protein was transformed to an active inhibitor of HIV-i :induced fusion. This transformation may result from liberation of the potent DP178 domain from a molecular clasp with the leucine zipper, DP107 domain.
For clarity of discussion, the invention will be described primarily for DP178 peptide inhibitors of HIV. However, the principles may be analogously applied to other viruses, both enveloped and nonenveloped, and to other non-viral organisms.
5.1. DP178 AND DP178-LIKE PEPTIDES
The DP178 peptide (SEQ ID:1) of the invention corresponds to amino acid residues 638 to 673 of the transmembrane protein gp4l from the HIV-liõ1 isolate, and has the 36 amino acid sequence (reading from amino to carboxy terminus):

NHi-YTSLIHSLIEESQNQQEKNEQ;ELLELDKWASLWNWF-COOH (SEQ ID:1) In addition to the full-length DP178 (SEQ ID:1) 36-mer, the peptides of the invention may include truncations of the DP178 (SEQ ID:1) peptide which exhibit antifusogenic activity, antiviral activity and/or the ability to modulate intracellular processes involving coiled-coil peptide structures. Truncations of DP178 (SEQ ID:1) peptides may comprise peptides of between 3 and 36 amino acid residues (i.e., peptides ranging in size from a tripeptide to a 36-mer polypeptide), as shown in Tables I and IA, below.
Peptide sequences in these tables are listed from amino (left) to carboxy (right) terminus. "X" may represent an amino group (-NH2) and "Z" may represent a carboxyl (-COOH) group. Alternatively, "X" may represent a hydrophobic group, including but not limited to carbobenzyl, dansyl, or T-butoxycarbonyl;
an acetyl group; a 9-f luorenylmethoxy-carbonyl (FMOC) group; or a covalently attached macromolecular group, including but not limited to a lipid-fatty acid conjugate, polyethylene glycol, carbohydrate or peptide group. Further, "Z" may represent an amido group; a T-butoxycarbonyl group; or a covalently attached macromolecular group, including but not limited to a lipid-fatty acid conjugate, polyethylene glycol, carbohydrate or peptide group. A preferred "X" or "Z" macromolecular group is a peptide group.

TABLE I
DP178 (SEO ID:l) CARBOXY TRUNCATIONS
X-YTS-Z
X-YTSL-Z
X-YTSLI-Z
X-YTSLIH-Z
X-YTSLIHS-Z
X-YTSLIHSL-Z
X-YTSLIHSLI-Z
X-YTSLIHSLIE-Z
X-YTSLIHSLIEE-Z
X-YTSLIHSLIEES-Z
X-YTSLIHSLIEESQ-Z
X-YTSLIHSLIEESQN-Z
X-YTSLIHSLIEESQNQ-Z
X-YTSLIHSLIEESQNQQ-Z
X-YTSLIHSLIEESQNQQE-Z
X-YTSLIHSLIEESQNQQEK-Z
X-YTSLIHSLIEESQNQQEKN-Z
X-YTSLIHSLIEESQNQQEKNE-Z
X-YTSLIHSLIEESQNQQEKNEQ-Z
X-YTSLIHSLIEESQNQQEKNEQE-Z
X-YTSLIHSLIEESQNQQEKNEQEL-Z
X-YTSLIHSLIEESQNQQEKNEQELL-Z
X-YTSLIHSLIEESQNQQEKNEQELLE-Z
X-YTSLIHSLIEESQNQQEKNEQELLEL-Z
X-YTSLIHSLIEESQNQQEKNEQELLELD-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDK-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKW-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKWA-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKWAS-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASL-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLW-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWN-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNW-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF-Z
The one letter amino acid code is used.
Additionally, "X" may represent an amino group, a hydrophobic group, including but not limited to carbobenzoxyl, dansyl, or T-butyloxycarbonyl; an acetyl group; a 9-fluorenylmethoxy-carbonyl (FMOC) group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

"Z" may represent a carboxyl group; an amido group; a T-butyloxycarbonyl group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

TABLE IA
DP178 (SEO ID:1) AMINO TRUNCATIONS

X-NWF-Z
X-WNWF-Z
X-LWNWF-Z
X-SLWNWF-Z
X-ASLWNWF-Z
X-WASLWNWF-Z
X-KWASLWNWF-Z
X-DKWASLWNWF-Z
X-LDKWASLWNWF-Z
X-ELDKWASLWNWF-Z
X-LELDKWASLWNWF-Z
X-LLELDKWASLWNWF-Z
X-ELLELDKWASLWNWF-Z
X-QELLELDKWASLWNWF-Z
X-EQELLELDKWASLWNWF-Z
X-NEQELLELDKWASLWNWF-Z
X-KNEQELLELDKWASLWNWF-Z
X-EKNEQELLELDKWASLWNWF-Z
X-QEKNEQELLELDKWASLWNWF-Z
X-QQEKNEQELLELDKWASLWNWF-Z
X-NQQEKNEQELLELDKWASLWNWF-Z
X-QNQQEKNEQELLELDKWASLWNWF-Z
X-SQNQQEKNEQELLELDKWASLWNWF-Z
X-ESQNQQEKNEQELLELDKWASLWNWF-Z
X-EESQNQQEKNEQELLELDKWASLWNWF-Z
X-IEESQNQQEKNEQELLELDKWASLWNWF-Z
X-LIEESQNQQEKNEQELLELDKWASLWNWF-Z
X-SLIEESQNQQEKNEQELLELDKWASLWNWF-Z
X-HSLIEESQNQQEKNEQELLELDKWASLWNWF-Z
X- IHSLIEESQNQQEKNEQELLELDKWASLWNWF-Z
X-LIHSLIEESQNQQEKNEQELLELDKWASLWNWF-Z
X-SLIHSLIEESQNQQEKNEQELLELDKWASLWNWF-Z
X-TSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF-Z
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWF-Z
The one letter amino acid code is used.
Additionally, -"X" may represent an amino group, a hydrophobic group, including but not limited to carbobenzoxyl, dansyl, or T-butyloxycarbonyl; an acetyl group; a 9-fluorenylmethoxy-carbonyl group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

"Z" may represent a carboxyl group; an amido group; a T-butyloxycarbonyl group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

The peptides of the invention also include DP178-like peptides. "DP178-like", as used herein, refers, first, to DP178 and DP178 truncations which contain one or more amino acid substitutions, insertions and/or deletions. Second, "DP-178-like" refers to peptide sequences identified or recognized by the ALLMOTI5, 107x178x4 and PLZIP search motifs described herein, having structural and/or amino acid motif similarity to DP178. The DP178-like peptides of the invention may exhibit antifusogenic or antiviral activity, or may exhibit the ability to modulate intracellular processes involving coiled-coil peptides. Further, such DP178-like peptides may possess additional advantageous features, such as, for example, increased bioavailability, and/or stability, or reduced host immune recognition.
HIV-1 and HIV-2 enveloped proteins are structurally distinct, but there exists a striking amino acid conservation within the DP178-corresponding regions of HIV-1 and HIV-2. The amino acid conservation is of a periodic nature, suggesting some conservation of structure and/or function. Therefore, one possible class of amino acid substitutions would include those amino acid changes which are predicted to stabilize the structure of the DP178 peptides of the invention. Utilizing the DP178 and DP178 analog sequences described herein, the skilled artisan can readily compile DP178 consensus sequences and ascertain from these, conserved amino acid residues which would represent preferred amino acid substitutions.
The amino acid substitutions may be of a conserved or non-conserved nature. Conserved amino acid substitutions consist of replacing one or more amino acids of the DP178 (SEQ ID:1) peptide sequence with amino acids of similar charge, size, and/or hydrophobicity characteristics, such as, for example, a glutamic acid (E) to aspartic acid (D) amino acid substitution. Non-conserved substitutions consist of replacing one or more amino acids of the DP178 (SEQ
ID:1) peptide sequence with amino acids possessing dissimilar charge, size, and/or hydrophobicity characteristics, such as, for example, a glutamic acid (E) to valine (V) substitution.
Amino acid insertions may consist of single amino acid residues or stretches of residues. The insertions may be made at the carboxy or amino terminal end of the DP178 or DP178 truncated peptides, as well as at a position internal to the peptide.
Such insertions will generally range from 2 to 15 amino acids in length. It is contemplated that insertions made at either the carboxy or amino terminus of the peptide of interest may be of a broader size range, with about 2 to about 50 amino acids being preferred. One or more such insertions may be introduced into DP178 (SEQ.ID:1) or DP178 truncations, as long as such insertions result in peptides which may still be recognized by the 107x178x4, ALLMOTI5 or PLZIP search motifs described herein, or may, alternatively, exhibit antifusogenic or antiviral activity, or exhibit the ability to modulate intracellular processes involving coiled-coil peptide structures.
Preferred amino or carboxy terminal insertions are peptides ranging from about 2 to about 50 amino acid residues in length, corresponding to gp4l protein regions either amino to or carboxy to the actual DP178 gp41 amino acid sequence, respectively. Thus, a preferred amino terminal or carboxy terminal amino acid insertion would contain gp4l amino acid sequences found immediately amino to or carboxy to the DP178 region of the gp4l protein.

Deletions of DP178 (SEQ ID:1) or DP178 truncations are also within the scope of the invention. Such deletions consist of the removal of one or more amino acids from the DP178 or DP178-like peptide sequence, with the lower limit length of the resulting peptide sequence being 4 to 6 amino acids.
Such deletions may involve a single contiguous or greater than one discrete portion of the peptide sequences. One or more such deletions may be introduced into DP178 (SEQ.ID:1) or DP178 truncations, as long as such deletions result in peptides which may still be recognized by the 107x178x4, ALLMOTI5 or PLZIP search motifs described herein, or may, alternatively, exhibit antifusogenic or antiviral activity, or exhibit the ability to modulate intracellular processes involving coiled-coil peptide structures.
DP178 analogs are further described, below, in Section 5.3.

5.2. DP107 AND DP107-LIKE PEPTIDES
Further, the peptides of the invention include peptides having amino acid sequences corresponding to DP107 analogs. DP107 is a 38 amino acid peptide which exhibits potent antiviral activity, and corresponds to residues 558 to 595 of HIV-1LAI transmembrane (TM) gp4l protein, as shown here:

(SEQ ID:25) In addition to the full-length DP107 (SEQ ID:25) 38-mer, the peptides of the invention may include truncations of the DP107 (SEQ ID:25) peptide which exhibit antifusogenic activity, antiviral activity and/or the ability to modulate intracellular processes involving coiled-coil peptide structures. Truncations of DP107 (SEQ ID:25) peptides may comprise peptides of between 3 and 38 amino acid residues (i.e., peptides ranging in size from a tripeptide to a 38-mer polypeptide), as shown in Tables II and IIA, below.
Peptide sequences in these tables are listed from, amino (left) to carboxy (right) terminus. "X" may represent an amino group (-NH2) and "Z" may represent a carboxyl (-COOH) group. Alternatively, "X" may represent a hydrophobic group, including but not limited to carbobenzyl, dansyl, or T-butoxycarbonyl;
an acetyl group; a 9-f luorenylmethoxy-carbonyl (FMOC) group; or a covalently attached macromolecular group, including but not limited to a lipid-fatty acid conjugate, polyethylene glycol, carbohydrate or peptide group. Further, "Z" may represent an amido group; a T-butoxycarbonyl group; or a covalently attached macromolecular group, including but not limited to a lipid-fatty acid conjugate, polyethylene glycol, carbohydrate or peptide group. A preferred õX" or "Z" macromolecular group is a peptide group.

TABLE II
DP107 (SEO ID:25) CARBOXY TRUNCATIONS
X-NNL-Z
X-NNLL-Z
X-NNLLR-Z
X-NNLLRA-Z
X-NNLLRAI-Z
X-NNLLRAIE-Z
X-NNLLRAIEA-Z
X-NNLLRAIEAQ-Z
X-NNLLRAIEAQQ-Z
X-NNLLRAIEAQQH-Z
X-NNLLRAIEAQQHL-Z
X-NNLLRAIEAQQHLL-Z
X-NNLLRAIEAQQHLLQ-Z
X-NNLLRAIEAQQHLLQL-Z
X-NNLLRAIEAQQHLLQLT-Z
X-NNLLRAIEAQQHLLQLTV-Z
X-NNLLRAIEAQQHLLQLTVW-Z
X-NNLLRAIEAQQHLLQLTVWQ-Z
X-NNLLRAIEAQQHLLQLTVWQI-Z
X-NNLLRAIEAQQHLLQLTVWQIK-Z
X-NNLLRAIEAQQHLLQLTVWQIKQ-Z
X-NNLLRAIEAQQHLLQLTVWQIKQL-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQ-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQA-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQAR-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARI-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARIL-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILA-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAV-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVE-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVER-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERY-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERYL-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERYLK-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERYLKD-Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQ-Z
The one letter amino acid code is used.
Additionally, "X" may represent an amino group, a hydrophobic group, including but not limited to carbobenzoxyl, dansyl, or T-butyloxycarbonyl; an acetyl group; a 9-fluorenylmethoxy-carbonyl (FMOC) group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.
"Z" may represent a carboxyl group; an amido group; a T-butyloxycarbonyl group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

TABLE IIA
DP178 (SEO ID:25) AMINO TRUNCATIONS

X-KDQ- Z
X-LKDQ- Z
X-YLKDQ- Z
X-RYLKDQ- Z
X-ERYLKDQ- Z
X-VERYLKDQ- Z
X-AVERYLKDQ- Z
X-LAVERYLKDQ- Z
X-ILAVERYLKDQ- Z
X-RILAVERYLKDQ- Z
X-ARILAVERYLKDQ- Z
X-QARILAVERYLKDQ- Z
X-LQARILAVERYLKDQ- Z
X-QLQARILAVERYLKDQ- Z
X-KQLQARILAVERYLKDQ- Z
X-IKQLQARILAVERYLKDQ- Z
X-QIKQLQARILAVERYLKDQ- Z
X-WQIKQLQARILAVERYLKDQ- Z
X-VWQIKQLQARILAVERYLKDQ- Z
X-TVWQIKQLQARILAVERYLKDQ- Z
X-LTVWQIKQLQARILAVERYLKDQ- Z
X-QLTVWQIKQLQARILAVERYLKDQ- Z
X-LQLTVWQIKQLQARILAVERYLKDQ- Z
X-LLQLTVWQIKQLQARILAVERYLKDQ- Z
X-HLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-QHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-QQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-AQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-EAQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-IEAQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-AI EAQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-RAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-LRAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-LLRAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-NLLRAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
X-NNLLRAIEAQQHLLQLTVWQIKQLQARILAVERYLKDQ- Z
The one letter amino acid code is used.

Additionally, "X" may represent an amino group, a hydrophobic group, including but not limited to carbobenzoxyl, dansyl, or T-butyloxycarbonyl; an acetyl group; a 9-fluorenylmethoxy-carbonyl group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.
"Z" may represent a carboxyl group; an amido group; a T-butyloxycarbonyl group; a macromolecular carrier.
group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

The peptides of the invention also include DP107-like peptides. "DP107-like", as used herein, refers, first, to DP107 and DP107 truncations which contain one or more amino acid substitutions, insertions and/or deletions. Second, "DP-107-like" refers to peptide sequences identified or recognized by the ALLMOTI5, 107x178x4 and PLZIP search motifs described herein, having structural and/or amino acid motif similarity to DP107. The DP107-like peptides of the invention may exhibit antifusogenic or antiviral activity, or may exhibit the ability to modulate intracellular processes involving coiled-coil peptides. Further, such DP107-like peptides may possess additional advantageous features, such as, for example, increased bioavailability, and/or stability, or reduced host immune recognition.
HIV-1 and HIV-2 enveloped proteins are structurally distinct, but there exists a striking amino acid conservation within the DP107-corresponding regions of HIV-1 and HIV-2. The amino acid conservation is of a periodic nature, suggesting some conservation of structure and/or function. Therefore, one possible class of amino acid substitutions would include those amino acid changes which are predicted to stabilize the structure of the DP107 peptides of the invention. Utilizing the DP107 and DP107 analog sequences described herein, the skilled artisan can readily compile DP107 consensus sequences and ascertain from these, conserved amino acid residues which would represent preferred amino acid substitutions.
The amino acid substitutions may be of a conserved or non-conserved nature. Conserved amino acid substitutions consist of replacing one or more amino acids of the DP107 (SEQ ID:25) peptide sequence with amino acids of similar charge, size, and/or hydrophobicity characteristics, such as, for example, a glutamic acid (E) to aspartic acid (D) amino acid substitution. Non-conserved substitutions consist of replacing one or more amino acids of the DP107 (SEQ
ID:25) peptide sequence with amino acids possessing dissimilar charge, size, and/or hydrophobicity characteristics, such as, for example, a glutamic acid (E) to valine (V) substitution.
Amino acid insertions may consist of single amino acid residues or stretches of residues. The insertions may be,made at the carboxy or amino terminal end of the DP107 or DP107 truncated peptides, as well as at a position internal to the peptide.
Such insertions will generally range from 2 to 15 amino acids in length. It is contemplated that insertions made at either the carboxy or amino terminus of the peptide of interest may be of a broader size range, with about 2 to about 50 amino acids being preferred. One or more such insertions may be introduced into DP107 (SEQ.ID:25) or DP107 truncations, as long as such insertions result in peptides which may still be recognized by the 107x178x4, ALLMOTI5 or PLZIP search motifs described herein, or may, alternatively,,exhibit antifusogenic or antiviral activity, or exhibit the ability to modulate intracellular processes involving coiled-coil peptide structures.
Preferred amino or carboxy terminal insertions are peptides ranging from about 2 to about 50 amino acid residues in length, corresponding to gp4l protein regions either amino to or carboxy to the actual DP107 gp41 amino acid sequence, respectively. Thus, a preferred amino terminal or carboxy terminal amino acid insertion would contain gp41 amino acid sequences found immediately amino to or carboxy to the DP107 region of the gp4l protein.

Deletions of DP107 (SEQ ID:25) or JP178 truncations are also within the scope of the invention. Such deletions consist of the removal of one or more amino acids from the DPI07 or DP107-like peptide sequence, with the lower limit length of the resulting peptide sequence being 4 to 6 amino acids.
Such deletions may involve a single contiguous or greater than one discrete portion of the peptide sequences. One or more such deletions may be introduced into DPI07 (SEQ.ID:25) or DP107 So truncations, as long as such deletions result in peptides which may still be recognized by the 107xl78x4, ALLMOTIS or PLZLP search motifs described herein, or may,:alternatively, exhibit antifusogenic or antiviral activity, or exhibit the ability to l5 modulate intracellular processes involving coiled-coil peptide structures.
DP107 and DP107 truncations are more fully described in:U.S. Patent No. 5,656,480, filed aanuary 27, 20 1995, DP107 analogs are further described, below, in Section 5.3.

5.3. DP107 and OP178 ANALOGS
25 Paptides corresponding to analogs of the DP178, DP178 truncations, DP107 and DP107 truncation sequences of the invention, described, above, in Sections 5.1 and 5.2 may be found in other viruses, including, for example, zion-HIV-1 1 enveloped viruses, 30 non-enveloped viruses and other non-viral organisms.
The term "analog", as used herein, refers to a peptide which is recognized or identified via the 107x178x4, ALLMOTI5 and/or PLZIP search strategies discussed below. Further, such peptides may exhibit 35 antifusogenic capability, antiviral activity, or the ability to modulate intracellular processes involving coiled-coil structures.
Such DP178 and DP107 analogs may, for example, correspond to peptide sequences present in TM proteins of enveloped viruses and may, additionally correspond to peptide sequences present in non enveloped and non-viral organisms. Such peptides may exhibit antifusogenic activity, antiviral activity, most particularly antiviral activity which is specific to the virus in which their native sequences are found, or may exhibit an ability to modulate intracellular processes involving coiled-coil peptide structures.
DP178 analogs are peptides whose amino acid sequences are comprised of the amino acid sequences of peptide regions of, for example, other (i.e., other than HIV-ILAI) viruses that correspond to the gp4l peptide region from which DP178 (SEQ ID:1) was derived. Such viruses may include, but are not limited to, other HIV-1 isolates and HIV-2 isolates.
DP178 analogs derived from the corresponding gp4l peptide region of other (i.e., non HIV-lI.AI) HIV-1 isolates may include, for example, peptide sequences as shown below.

NH2-YTNTIYTLLEESQNQQEKNEQELLELDKWASLWNWF-COQH (DP-185; SEQ
ID:3);

NH2-YTGIIYNLLEESQNQQEKNEQELLELDKWANLWNWF-COOH (SEQ ID:4);
NH2-YTSLIYSLLEKSQIQQEKNEQELLELDKWASLWNWF-COOH (SEQ ID:5).

SEQ ID:3 (DP-185), SEQ ID:4, and SEQ ID:5 are derived from HIV-1SF2, HIV-lRF, and HIV-lmQ,1 isolates, respectively. Underlined amino acid residues refer to those residues that differ from the corresponding position in the DP178 (SEQ ID:1) peptide. One such DP178 analog, DP-185 (SEQ ID:3), is described in the Example presented in Section 6, below, where it is demonstrated that DP-185 (SEQ ID:3) exhibits antiviral activity. The DP178 analogs of the invention may also include truncations, as described above. Further, the analogs of the invention modifications such those described for DP178 analogs in Section 5.1., above.
It is preferred that the DP178 analogs of the invention represent peptides whose amino acid sequences correspond to the DP178 region of the gp4l protein, it is also contemplated that the peptides of the invention may, additionally, include amino sequences, ranging from about 2 to about 50 amino acid residues in length, corresponding to gp4l protein regions either amino to or carboxy to the actual DP178 amino acid sequence.
Striking similarities, as shown in FIG. 1, exist within the regions of HIV-1 and HIV-2 isolates which correspond to the DP178 sequence. A DP178 analog derived from the HIV-2NII.IZ isolate has the 36 amino acid sequence (reading from amino to carboxy terminus):
NH2-LEANISQSLEQAQIQQEKNMYELQKLNSWDVFTNWL-COOH (SEQ ID:7) Table III and Table IV show some possible truncations of the HIV-2Nmz DP178 analog, which may comprise peptides of between 3 and 36 amino acid residues (i.e., peptides ranging in size from a tripeptide to a 36-mer polypeptide). Peptide sequences in these tables are listed from amino (left) to carboxy (right) terminus. "X" may represent an amino group (-NH2) and "Z" may represent a carboxyl (-COOH) group.
Alternatively, "X" may represent a hydrophobic group, including but not limited to carbobenzyl, dansyl, or T-butoxycarbonyl; an acetyl group; a 9-fluorenylmethoxy-carbonyl (FMOC) group; or a covalently attached macromolecular group, including but not limited to a lipid-fatty acid conjugate, polyethylene glycol, carbohydrate or peptide group.
Further, "Z" may represent an amido group; a T-butoxycarbonyl group; or a covalently attached macromolecular group, including but not limited to a lipid-fatty acid conjugate, polyethylene glycol, carbohydrate or peptide group. A preferred "X" or "Z"
macromolecular group is a peptide group.

TABLE III
HIV-2Nm DP178 analog carboxy truncations.
X-LEA-Z
X-LEAN-Z
X-LEANI-Z
X-LEANIS-Z
X-LEANISQ-Z
X-LEANISQS-Z
X-LEANISQSL-Z
X-LEANISQSLE-Z
X-LEANISQSLEQ-Z
X-LEANISQSLEQA-Z
X-LEANISQSLEQAQ-Z
X-LEANISQSLEQAQI-Z
X-LEANISQSLEQAQIQ-Z
X-LEANISQSLEQAQIQQ-Z
X-LEANISQSLEQAQIQQE-Z
X-LEANISQSLEQAQIQQEK-Z
X-LEANISQSLEQAQIQQEKN-Z
X-LEANISQSLEQAQIQQEKNM-Z
X-LEANISQSLEQAQIQQEKNMY-Z
X-LEANISQSLEQAQIQQEKNMYE-Z
X-LEANISQSLEQAQIQQEKNMYEL-Z
X-LEANISQSLEQAQIQQEKNMYELQ-Z
X-LEANISQSLEQAQIQQEKNMYELQK-Z
X-LEANISQSLEQAQIQQEKNMYELQKL-Z
X-LEANISQSLEQAQIQQEKNMYELQKLN-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNS-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSW-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSWD-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSWDV-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSWDVF-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSWDVFT-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSWDVFTN-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSWDVFTNW-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
The one letter amino acid code is used.
Additionally, -"X" may represent an amino group, a hydrophobic group, including but not limited to carbobenzoxyl, dansyl, or T-butyloxycarbonyl; an acetyl group; a 9-fluorenylmethoxy-carbonyl (FMOC) group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

"Z" may represent a carboxyl group; an amido group; a T-butyloxycarbonyl group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.
-TABLE IV
HIV-2ND DP178 analog amino truncations.
= X-NWL-Z
X-TNWL-Z
X-FTNWL-Z
X-VFTNWL-Z
X-DVFTNWL-Z
X-WDVFTNWL-Z
X-SWDVFTNWL-Z
X-NSWDVFTNWL-Z
X-LNSWDVFTNWL-Z
X-KLNSWDVFTNWL-Z
X-QKLNSWDVFTNWL-Z
X-LQKLNSWDVFTNWL-Z
X- ELQKLNSWDVFTNWL-Z
X-YELQKLNSWDVFTNWL-Z
X-MYELQKLNSWDVFTNWL-Z
X-NMYELQKLNSWDVFTNWL-Z
X-KNMYELQKLNSWDVFTNWL-Z
X-EKNMYELQKLNSWDVFTNWL-Z
X-QEKNMYELQKLNSWDVFTNWL-Z
X-QQEKNMYELQKLNSWDVFTNWL-Z
X- IQQEKNMYELQKLNSWDVFTNWL-Z
X-QIQQEKNMYELQKLNSWDVFTNWL-Z
X-AQIQQEKNMYELQKLNSWDVFTNWL-Z
X-QAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-EQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-LEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-SLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-QSLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-SQSLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-ISQSLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-NISQSLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-ANISQSLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-EANISQSLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
X-LEANISQSLEQAQIQQEKNMYELQKLNSWDVFTNWL-Z
The one letter amino acid code is used.
Additionally, "X" may represent an amino group, a hydrophobic group, including but not limited to carbobenzoxyl, dansyl, or T-butyloxycarbonyl; an acetyl group; a 9-fluorenylmethoxy-carbonyl (FMOC) group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

"Z" may represent a carboxyl group; an amido group; a T-butyloxycarbonyl group; a macromolecular carrier group including but not limited to lipid-fatty acid conjugates, polyethylene glycol, or carbohydrates.

DP178 and DP107 analogs are recognized or identified, for example, by utilizing one or more of the 107xl78x4, ALLMOTI5 or PLZIP computer-assisted search strategies described and demonstrated, below, in the Examples presented in Sections 9 through 16 and 19 through 25. The search strategy identifies additional peptide regions which are predicted to have structural and/or amino acid sequence features similar to those of DP107 and/or DP178.
The search strategies are described fully, below, in the Example presented in Section 9. While this search strategy is based, in part, on a primary amino acid motif deduced from DP107 and DP178, it is not based solely on searching for primary amino acid sequence homologies, as such protein sequence homologies exist within, but not between major groups of viruses. For example, primary amino acid sequence homology is high within the TM protein of different strains of HIV-1 or within the TM protein of different isolates of simian immunodeficiency virus (SIV).
Primary amino acid sequence homology between HIV-1 and SIV, however, is low enough so as not to be useful.
It is not possible, therefore, to find peptide regions similar to DP107 or DP178 within other viruses, or within non-viral organisms, whether structurally, or otherwise, based on primary sequence homology, alone.
Further, while it would be potentially useful to identify primary sequence arrangements of amino acids based on, for example, the physical chemical characteristics of different classes of amino acids rather than based on the specific amino acids themselves, such search strategies have, until now, proven inadequate. For example, a computer algorithm designed by Lupas et al. to identify coiled-coil propensities of regions within proteins (Lupas, A., et al., 1991 Science 252:1162-1164) is inadequate for identifying protein regions analogous to DP107 or DP178.
Specifically, analysis of HIV-1 gp160 (containing both gp120 and gp4l) using the Lupas algorithm does not identify the coiled-coil region within DP107. it does, however, identify a region within DP178 beginning eight amino acids N-terminal to the start of DP178 and ending eight amino acids from the C-terminus. The DP107 peptide has been shown experimentally to form a stable coiled coil. A search based on the Lupas search algorithm, therefore, would not have identified the DP107 coiled-coil region.
Conversely, the Lupas algorithm identified the DP178 region as a potential coiled-coil motif. However, the peptide derived from the DP178 region failed to form a coiled coil in solution.
A possible explanation for the inability of the Lupas search algorithm to accurately identify coiled-coil sequences within the HIV-1 TM, is that the Lupas algorithm is based on the structure of coiled coils from proteins that are not structurally or functionally similar to the TM proteins of viruses, antiviral peptides (e.g. DP107 and DP178) of which are an object of this invention.
The computer search strategy of the invention, as demonstrated in the Examples presented below, in Sections 9 through 16 and 19 through 25, successfully identifies regions of proteins similar to DP107 or DP178. This search strategy was designed to be used with a commercially-available sequence database package, preferably PC/Gene.
A series of search motifs, the 107x178x4, ALLMOTI5 and PLZIP motifs, were designed and engineered to range in stringency from strict to broad, as discussed in this Section and in section 9, with 107x178x4 being preferred. The sequences identified via such search motifs, such as those listed in Tables V-XIV, below, potentially exhibit antifusogenic, such as antiviral, activity, may additionally be useful in the identification of antifusogenic, such as antiviral, compounds, and are intended to be within the scope of the invention.
Coiled-coiled sequences are thought to consist of heptad amino acid repeats. For ease of description, the amino acid positions within the heptad repeats are sometimes referred to as A through G, with the first position being A, the second B, etc. The motifs used to identify DP107-like and DP178-like sequences herein are designed to specifically search for and identify such heptad repeats. In the descriptions of each of the motifs described, below, amino acids enclosed by brackets i.e., [], designate the only amino acid residues that are acceptable at the given position, while amino acids enclosed by braces, i.e., {}, designate the only amino acids which are unacceptable at the given heptad position. When a set of bracketed or braced amino acids is followed by a number in parentheses i.e., (), it refers to the number of subsequent amino acid positions for which the designated set of amino acids hold, e.g, a (2) means "for the next two heptad amino acid positions".
The ALLMOTI5 is written as follows:
{CDGHP}-{CFP}(2)-{CDGHP}-{CFP}(3)-{CDGHP}-{CFP}(2)-{CDGHP}-{CFP}(3)-{CDGHP}-{CFP}(2)-{CDGHP}-{CFP}(3)-{CDGHP}-{CFP}(2)-{CDGHP}-{CFP}(3)-{CDGHP}-{CFP}(2)-{CDGHP}-{CFP}(3)-Translating this motif, it would read: "at the first (A) position of the heptad, any amino acid residue except C, D, G, H, or P is acceptable, at the next two (B,C) amino acid positions, any amino acid residue except _-, F, or P is acceptable, at the fourth heptad position (D), any amino acid residue except C, D, G, H, or P is acceptable, at the next three (E, F, G) amino acid positions, any amino acid residue except C, F, or P is acceptable. This motif is designed to search for five consecutive heptad repeats (thus the repeat of the first line five times), meaning that it searches for 35-mer sized peptides. It may also be designed to search for 28-mers, by only repeating the initial motif four times. With respect to the ALLMOTI5 motif, a 35-mer'search is preferred. Those viral (non-bacteriophage) sequences identified via such an ALLMOTI5 motif are listed in Table V, below, at the end of this Section. The viral sequences listed in Table V potentially exhibit antiviral activity, may be useful in the the identification of antiviral compounds, and are intended to be within the scope of the invention. In those instances wherein a single gene exhibits greater than one sequence recognized by the ALLMOTI5 search motif, the amino a cid residue numbers of these sequences are listed under "Area 2", Area 3", etc. This convention is used for each of the Tables listed, below, at the end of this Section.
The 107x178x4 motif is written as follows:
[EFIKLNQSTVWY]-{CFMP}(2)-[EFIKLNQSTVWY]-{CFMP}(3)-[EFIKLNQSTVWY]-{CFMP}(2)-[EFIKLNQSTVWY]-{CFMP}(3)-(EFIKLNQSTVWY]-{CFMP}(2)-[EFIKLNQSTVWY]-{CFMP}(3)-[EFIKLNQSTVWY]-{CFMP}(2)-[EFIKLNQSTVWY]-{CFMP}(3)-Translating this motif, it would read: "at the first (A) position of the heptad, only amino acid residue E, F, I, K, L, N, Q, S, T, V, W, or Y is acceptable, at the next two (B,C) amino acid positions, any amino acid residue except C, F, M or P
is acceptable, at the fourth position (D), only amino acid residue E, F, I, K, L, N, Q, S, T, V, W, or Y is acceptable, at the next three (E, F, G) amino acid positions, any amino acid residue except C, F, M or P
is acceptable. This motif is designed to search for -four consecutive heptad repeats (thus the repeat of the first line four times), meaning that it searches for 28-mer sized peptides. It may also be designed to search for 35-mers, by repeating the initial motif five times. With respect to the 107x178x4 motif, a 28-mer search is preferred.
Those viral (non-bacteriophage) sequences identified via such a 107x178x4 motif are listed in Table VI, below, at the end of this Section, with those viral (non-bacteriophage) sequences listed in Table VII, below at the end of this Section, being preferred.
The 107x178x4 search motif was also utilized to identify non-viral procaryotic protein sequences, as listed in Table VIII, below, at the end of this Section. Further, this search motif was used to reveal a number of human proteins. The results of this human protein 107x178x4 search is listed in Table IX, below, at the end of this Section. The sequences listed in Tables VIII and IX, therefore, reveal peptides which may be useful as antifusogenic compounds or in the identification of antifusogenic compounds, and are intended to be within the scope of the invention.
The PLZIP series of motifs are as listed in FIG.
19. These motifs are designed to identify leucine zipper coiled-coil like heptads wherein at least one proline residue is present at some predefined distance N-terminal to the repeat. These PLZIP motifs find regions of proteins with similarities to HIV-1 DP178 generally located just N-terminal to the transmembrane anchor. These motifs may be translated according to the same convention described above. Each line depicted in FIG. 19 represents a single, complete search motif. "X" in these motifs refers to any amino acid residue. In instances wherein a motif contains two numbers within parentheses, this refers to a variable number of amino acid residues. For example, X (1,12) is translated to "the next one to twelve amino acid residues, inclusive, may be any amino acid".
Tables X through XIV, below, at the end of this Section, list sequences identified via searches conducted with such PLZIP motifs. Specifically, Table X lists viral sequences identified via PCTLZIP, P1CTLZIP and P2CTLZIP search motifs, Table XI lists viral sequences identified via P3CTLZIP, P4CTLZIP, P5CTLZIP and P6CTLZIP search motifs, Table XII lsts viral sequences identified via P7CTLZIP, P8CTLZIP and P9CTLZIP search motifs, Table XIII lists viral sequences identified via P12LZIPC searches and Table XIV lists viral sequences identified via P23TLZIPC
search motifs The viral sequences listed in these tables represent peptides which potentially exhibit antiviral activity, may be useful in the identification of antiviral compounds, and are intended to be within the scope of the invention.
The Examples presented in Sections 17, 18, 26 and 27 below, demonstrate that viral sequences identified via the motif searches described herein identify substantial antiviral characteristics. Specifically, the Example presented in Section 17 describes peptides with anti-respiratory syncytial virus activity, the Example presented in Section 18 describes peptides with anti-parainfluenza virus activity, the Example presented in Section 26 describes peptides with anti-measles virus activity and the Example presented in Section 27 describes peptides with anti-simian immunodeficiency virus activity.
The DP107 and DP178 analogs may, further, contain any of the additional groups described for DP178, above, in Section 5.1. For example, these peptides may include any of the additional amino-terminal groups as described above for "X" groups, and may also include any of the carboxy-terminal groups as described, above, for "Z" groups.
Additionally, truncations of the identified DP107 and DP178 peptides are among the peptides of the invention. Further, such DP107 and DP178 analogs and DP107/DP178 analog truncations may exhibit one or more amino acid substitutions, insertion, and/or deletions.
The DP178 analog amino acid substitutions, insertions and deletions, are as described, above, for DP178-like peptides in Section 5.1. The DP-107 analog amino acid substitutions, insertions and deletions are also as described, above, for DP107-like peptides in Section 5.2.
Tables XV through XXII, below, present representative examples of such DP107/DP178 truncations. Specifically, Table XV presents Respiratory Syncytial Virus F1 region DP107 analog carboxy truncations, Table XVI presents Respiratory Syncytial Virus F1 region DP107 analog amino truncations, Table XVII presents Respiratory Syncytial Virus F1 region DP178 analog carboxy truncations, Table XVIII presents Respiratory Syncytial Virus F1 region DP178 analog amino truncations, Table XIX
presents Human Parainfluenza Virus 3 F1 region DP178 analog carboxy truncations, Table XX presents Human Parainfluenza Virus 3 F1 region DP178 analog amino truncations, Table XXI presents Human Parainfluenza Virus 3 F1 region DP107 analog carboxy truncations and Table XXII presents Human Parainfluenza Virus 3 F1 region DP107 analog amino truncations. Further, Table XXIII, below, presents DP107/DP178 analogs and analog truncations which exhibit substantial antiviral activity. These antiviral peptides are grouped according to the specific virus which they inhibit, including respiratory syncytial virus, human parainfluenza virus 3, simian immunodeficiency virus and measles virus.

TABLE V

FOR ALL VIRAL (NON-BACTERIOPHAGE) PROTEINS

<~ <oo w r4 Z Z Z Z Z Z

Z~C rL 7_ rL

u u u u u u ZL ZZ ? ZL 2 << F
N N N N N N N N
~ aC 6G BL CL ~ OC OC OL
- U U U U U U U :J
< < < < < < < <
m u~i a w LU
0o~ 0 6 a.. b a. d Gam. a.. Yom.

Y Y Y Y Y Y Y
a ~ r A ffT .rj = f~ r i.Y 0 0 0 0 0 0 0 0 < x x x- s x U U U > > U U
U U U U U
U U UUUU
UUU

u c p y<

~. ~~. 6 6 6 d 6I6 TABLE VI

FOR ALL VIRAL (NON-BACTERIOPHAGE) PROTEINS

TABLE VII

(PREFERRED VIRAL SEQUENCES) (~III~IIiI~i) IIIII Illill it III
aillll II I IIIII
~Igli III
~PIIIIIIII
1=111 I it I I I ri 3SS15 9SS7u aaamaa111llloa"
Z Z Z!7.~7 I'L IT_ 2 Z
`IIwI~~^, - - -1~hf~i~lyi~h N >- S
i <.clc _c1: c~_cE ==

ooooiooo~oo~~~~~
N N N N N N N N N Y Y Y Y Y
}t ~ f J J J J J J J j U U U U
C U UYUIU UU UUU QOGOQG QC
< > > > >> > > > 3 3 3 3 3 iiiL1 ` S W V L< W < M < N y N -v ` ~ ~ Z ~ F.= ¾j1 ~ cj ~ ~ p p~
= Z{y{ 000 <Oi.~PPQ
~21UI Q). 00) C <<<<
So sc~~uu.ucccon ~V ~1 N N N ~V >N " .>tz >z=
Y > > > > > > > r3 3 3 3~
v 5'S~~S'>S5>oao`ao TABLE VIII

FOR ALL PROCARYOTIC PROTEINS

A

b P
T P N b -b r P P
_ _ ~ ~O r ev ^ b - ~ ^ P r ~ b r O r r~ r P~-uu r>

N X U N a N N N N N N U U
MOMS H H H H F- V
N U N N -n -N N Y. U 1 e O O J p J <
a U n~. U U 5 U U U U U U U
e`. mU-3mu immmmmmm <uum W>~~zzz~bU`~UJ Z
{Y ZL 4 C W W W W u7 u7 Y7 W W W W
O 0 h 0 0 0 0 0 0 0 0 0 o0cz0oo0 0 A. A. GOGA.OGeA.A.cA 0-o.
2 d C d 0 0 0 0 0 0 0 O C U 0 =0 P% -Y YY YYYYYY 0t.
b Y n Y r r o o r ,+ a r O r UUUUUUUUUUU
U U U U U U U U L):U U U U u r F 1 t- ~= r= r= t f. r= Ir-uuuu N N N N ...) t~4UU~uUV<~~U
67 fn .+ i0 H 0 ;111 m O m m m m< U V {Va7 r d n. n. n. n.-Mt n. n.s n. t a nal TABLE IX

FOR ALL HUMAN PROTEINS

= P
r P =p b n T

N r 4. -<
O O
H O
N O
~! Q
b e -------- ------------ ---N f N n N O
n O
N n P O r s o 4 $
r S o o H a tln S P
- o a ri~~a a _- _ T b N b M r f ~ H
{t. ~i n n n r ~ H H h NNp n ' pp~ O~ tl ~~ O O ~ P O P N A N O n P ~ b N SC ~ b~ G O N
- -- O O O =p O ~ =O !~ ~ O N P
< O N O -^ ^ n n t~ .p N -- 0 V- N r ~. n n-- P ~.~ tl~_ b =~ O .p Otl N h~ n- P er "~ N a rbi wPi ~ o O N .p N P
O D
- tl n N H b b P n = S p O f tl o P v. .== V_ c n ^- N^ N r r P P--- ^ P_ =p n_ f~ b f f P N b .p .p Q .p r h P N n^ N' _ r1 d b C yf ptlp 1~ T P o ~f Zr - .D h N N f a a c v r > Ã w w < ? v VUv m m <dl'.'ay z N =U- V <jai ww6wz 5. ~, u`1F~w~: xa<dPI maa 1f;!IiJ0if {yy ~~ yOO6Ca o zZ o <,=; ~ t~y' a 1 N T p x }q Y. Zy. 6u. z JUU SyO O 0 - t-! << vv O ~aC p+1 Y L O p a~ C v~~ O o. UV
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p V a ae oc of y~- > "p car x"'y~a~ J=tlo C~ty/~.
f W! N i~ V L > ~=' (y{~ 41 1t~ L S L {yy~ N .~.j (..l v N ^ W 4 < 0~ p L) a > ti p io a C < U O O O C U W J<_ C w- N w w o ~r V o' V
~w KAY 0 <õ7~ z ~ 6 '9 O JO <<no F. o r C'-=t-F.}~~Z OJ~pOatui tWi ttyJx 00d ST^U..y~U~~aSSai < r aGaGK 10u!'~oGaa~ E'o~ uc~`~` aaa"'< :8 o J0000-- b<tw{y< e~ O< =SS n.w H
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c4 a z xo= SXT US xS < _ ~+~t)Z a.q. U w,,,y ~q V <,1 CC pCr7 eWZ>
0 ~i00 nC. nom.
Z~ ~.n .n v~J= qy tyy a'06 n. u6~ Sy F =/~U ZV ZV ~~U UyJ {juUUC.a l < <
V _ < <
(7 5 F S '^ J ({{{~iiJ~~.. ({~ ~SSS} 1p~ 9 9 ^ < p~~Zj+, =~yN~ji ~=~yyyj+~gyj .[NY~J ^Yy~ p, q d d G= tf. d d 6 L L d b d 6 d d d d 6 d 6 6 d d b L B. 6 d d 6 O. 6 d 6 d d 6 6 6 6 d d d d 6 d 4 6 d 6 0 a 6 +~ p P I I I I se ~ oa .IopJ f --------- ------saa I I o~do }~ I I j l I I pay b _ N
r n p ' ~ N O= N p -O= b 0 0 ~` T ~ ~ P
h G P p I~' N =^ o. a o r o p a r r c ~ ^ R _ .o 1= - - ^ INI -o ^ H o h N ^ N tl O ~_ A õ N p p = O p P r P h T T^~~ h p~- =n =Y pN. N d O N fV C O= O N-~- =f ,n q p N Q~ N Q~ p N ^~ b N `^ O ~ e P =n n O N ^ I~ a =E - - = I^ ~ n = T ^ a a O - r O~ p _- -n n p b =~ - n N H p n~^ b H f n- ='1 b - r1 ~ b =a h ~'õ P~ appp ybj N N^
w ~ P^ f o- N N b^ - tl Q n~ N^ n b_ n N- a b Q =O H N C ^lT, b H
Ztit to - ~ r _2 ty 4 ry H = _ ~ tJ fti F
Om T F.. 4 .~dta O V `F,4ry- 2 u Fj ti < O t:7 V u1 h- "' F ~f<w[ C' ^ V O G TJ' y a m F <

=9 hEp.. to ?rn Wm t]5 py<p G V ,..~..1 =~p O==TVj v W y v v K N W 2= < a <4 p<
4' u U r.0 N co ?Oj u m i FF++
F upC~ gq''u'T~' j .W.)~ U{yp <tn O
O OCR m y!_u N U f N L=' U z, =l5 O O T' ~:.~ O L LL! U ~+ Y Z OS !~ C1 I-t~~'.
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og UU V uF V 77f_''. y<,z HIVõ N Ctt7 W W VyhK V Z GLLj Jy U,~,~,==~~(~,)<~~==
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u_uuuu a ~
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.= i N ~ ~ N h H N m ~ ~ U ¾ ~= ~ ~ O W ~ W ~, ~ < ~ ~ F w to N ~ ~ ~ ~ < 4~
4' ~ O O S =.7 .~ > ~' 44 U N N Z U y U UY - ZZZ U 4 f{i_I < OU I,{~ p FU N .{~ZiF.,.
~FF~FOOr W NOS ~~.~OO'~'J~OQ GFq DO U~I 4oG1OO P~ ~Z~"=< W
o ~m<e <uo~~~, co4~-~oYYca4'm4'~wc~i30, ~F~o~xFc}ioo~~
- U m V U U U U u U U V U 4 F~ I"' F` La U F~ ..] F U V F .d a u u u 2 u u ~ V
(~j V V~~ V y V V~ V U V~ V V U U U
w F qQ ~=1 m a s- n- m o c t t I
:7 < F-~F"u.044N C7 0...1 X ' ...l d' I < I
u~c<iuvv~mmmmuc~ucecacocoo c3aoccc~zt~
uuuuuuuu uuuuuuuvuuuu,I8o3ogo ~' ' ' ' ' = 4 4 4 4 4 4 4 4 4 4 6 4 4 4 4 4 4 4 4 U U U U U U u U U V U
U U V U ~ U ~~ UU (~ UU U
4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 d y U y y y = P a O p P N

P =
f b Ic ^ ~ ^ O O
^O n b b ^
.Q - O Q ^ - - ~ O b q ^
N ry ry H
O ^~ N ' P N P P =-=+ 4 P ~ =OO 0 ~_ eV p~o o. =n r~.r tl P P N =~-= N n S =/= - N ~ 6- ~ õ - n r0 - N N N A V o ^ n - n - - ~ ^ O ^ ^
O Or O n r ~' O p= P P=^ f n Q~ n T n- N N N O' P P N N P N P Or Or p^-- - - b N P r - - ~ . P P - - - - - ^ ~ O ePV b b - r 0 = r T T ,r ? O f ~ .1^
n pT - = N N N
--~ r N H N f~ P b- a~ =O O N =1 n b~ N
n N v< f N N -- n n Z w w h O O <- U
0 f0-.,,wF a ~~ `1' <<<= Hw o~ ltt v~r hq UU~rl~mmaO,r HZ Q
FF~7~~~<~~~1"" uUul1mmmm m~ Qmgga~ ~ ~ =~~
m Z~vOi ZN, V ON
(~7n W Fp0 ~ ~nr ~a0 S~mr<7 <y r<7 j ty NO
y VF s W _: << - 22x 5 c~u nib UaU
Qn p a W t~iU ~tr UHN {y_a ""Va1U-bbb K- KUi to a"3 o~ O 1O..w eCGUw =nn 22K~Z pa0.-..1 ZN Z U Ow F U< aa00woOdw <O t:'O w- -v <S~ UaeY
Oce?yldn o Z6 UH~aaay ooUbbbbbbb o er..:''Wz ~a~-w a oI gqooC o_w (Qa]sa y {{yyw y1y~~{yt~~{{yy~~ yyJ~ {{t~~{i~~z{F
p y y w .] y E .Q !~ r U O U F" < b Z N O F F a F F 1-r F F F~ F.. m F U 19 E U ~~ b U wbb'~' m aeju~io EuzUO3yo UUlueoeapa e cK eoca ~'+~w=xS<<1<~ 21 U t=t~ U U ~U e. aaaaaa ap~aF <
S'o yo4 o 2S-(t~ ~`PoooVO06 u w .. <vy < z ooo aaa (zo'o~a`a~` o~t 5y-g- h- 20Fpg69pb6QS4bpsi5 Qgb ^~mwF-I .
O F K 4' a 4 V ~+ 1<L ~Zj U U p 8 N x n L Vr Oi t~ << G O G C C C O ~' F 1Q.
L~ S U y e `
U; uf Q i m m Z Z U= GO t w y {{I b H U U m m m m m m m (7 m E= < rnr H
uI~G~a V~o~iIVt14~I~~ Us.~ FU ou5uglug1lguuSqWw ug5zyt Y~ d9 Jj a[~~~~Qa ~zQ <~~ZS~ U ~~itQQ V ~~~"~U~ N~~~yõ~FGFFFF`U9` Fw~" c~~ yQ <
<C.Ir r1 ~ZZZ"q ~..1 w~~~ ~0~~} ~~xw,,,~wmm~ooooooo QC.>Q a. a~.yw tl m~+ 0< Q y W w W W W w wely a'B 7 `Z~Z` bcG~l -.~g~ F zzo ~7 c e Ip~Ity~~rsos<~poCiLRitidc"~~`c7 c ~c ~ qU
~yZ<kQ
= 21u Oo44 "'Ã'^uzSwwuzi'O aFS-ya .~b '~'`'c ~Z2 a(zr < v ~ yee ~ x< pc~~ o w o~~~u a v z aooo b~~o U~ ~d > 6.> <~p3 ~adda l y 1-. oo x~
W ~Q~W 1N~000< OOmmLHHH.,,,,716 ~yy jy 5< U3O pqU,,000 ]! W U 6.> Y. ..1 4. u. u. LL u. 4r 4. L O W 4. Y. pa F W a y V 2 V V V V V
V V V V C7 (~ F ==] _7 ...1 < ..1 ~1 ~=1 W e yl {<t~ t 1 1 1 1 1 1 IT
V U~ F~~ `<<<< V (õ/ < m V .'S. U ~ y z O N a N~ ~ N ~" y N N 1 1 I
u~la7www c~3JH b pcC pT~Z~Z <S vy~ as n ~= aaaaaayyydayyae.d oa.ad Sr~y~~"=~~C~i= 2-a^~< 7~tccm~~cmc77~mmmmmfFA
aaao~'.aaanv.&~id. cO.aaaaady00007 C7 C~C~ V VC7 C~
a a a a a a a a a a a a a a a b tl N
------------------------------ -------tl Q

- - - - - - - - - - - - - - - - - - - - - - -N n --------------b N
b b b N
-----------------------------b N O
O ~ b @ N O Q
Y n ~
Y N N b 7 tl N
T N T
--------------tl . N
_ O p ^
- - - - - - - - - - - - - - - - - - - - -^ N tl b - b ^ - - - N O N
~ B tl N T A
r b tl O
------------------ - - N @ M
tl N b W
~ ^~~ ry n 4 a n n q N
N Q Q S O O f ~O
-----------------= _ - N
..' h^ N tl O - tl ^ N
f @
@ N N Q -- b n b n b n - ~ !1!!
e !
a~
o B P @ tl t~ @ h e~ p tl tl~ N N N
^ n T T- '^ O - N N r - N S N Q h P b= O r r tl n r O tl tl N -d a h N of p _ ~O N f O b^ O- O N n P Q ^= N 4 t~ tl- tl -? C. e.l nTl Q - n n n^ f` f~ ^ Q-~O N N N N t~ N N= P
n r A N tl- - - - - b -~ b~ b Q- N O n h~ n O f~V vOp N N@ n1 O~~ b A
r@ - - - b r fV N d n H O MI
w b n wf W N ~ - .~ .,. P OO

U << u~
y V L) Ow l: ~{y K 0 oL~~0 d y W w:9 <2 .Ti eOCNOO ? 9 p y ~Oa < LCd 90 O aS uI11.
-Z 6: v ~o ~~a t2 e ~y w v ^p s Ã< d<00 `~<waccZZ Q {y~{y 0 Li F_6 W ~. <,wZO0tUUyr z eyy f<, d yo'?py Og` $oG eC
96 yO ~~ VF p W
~{ fgiJ .~~.= ~Ot! $0.~ O yN' 7< ~F{ W W Z Z L 0. L U ~i {~~y71f' F- 0.
H L V d d h+ ? 4) b Q N uT3 N ` [VVV'- o 1Ua~ y _ y <y <y u b o I F ou l~ Sa y < iihlfl < ~a4 Ua F= w~ Om y of t-yss ~ IH < W 0. vl O. O W ~G p~

Vj O y UZ~ .t H ,JAS ~j OOUpaaC~P7c' Wy ~'~NFk+U
O~UL2<U !~v~ivUi~ azU O JUy y U d q OQ4p ¾p5 ~.t {{~~o {<y {Q~ O VO pxJ t ~> > N<V zua. U
4 @ w SyNa jF+p .7y Sl+x (1 ~~iz ~ Z~U OY<v~y Z z~~ F ~QO W W yp ~4y u~ u~f U
~ ~O a0.}
Opl^F+aa ~LL <?.~$3 Wa~a r% oa,~ 6N.a.4 S X a~~' -+
000. 8~y~Z YZZyyZ O W y,,{ o. uf t+'3 y - ~ ~.. O F O V x ~~~..11! t- K 0 0 ~ w W
nUU ~6: H-~~c f;,=FF Vz;'CWO ai 0700 V 2 x U sty O U cG G-<< W Z @I v~ NI L~ N N VLJ t0.if y<
! i i i ~ 2 i = 0. 4 0. 0. 6. 0.

- - i +i V

O
O
SI
< !1 O
M
D l O
O'O

P
~~ z^

z ov n. 4 WW~

z e U~ Y U Y
UU W +.:z = = w zm -of z q I- is C7 ~=~''..O
E <
g gg U_U U
ad K
t pp2Up[.N. aN
g UViC ~~Y

a s a a a a n t t~ U U U U
o zzz~zis O C G N IM N N
~ zzzz e zz z ~~zzzz 228 ~1~fO
a aaaaeN.=,nN.

TABLE X

Search Results Summary for PCTLZIP, P 1 CTLZIP, and P2CTLZIP Motifs TABLE XI

Search Results Summary for P3 CTLZIP, P4CTL;ZI P, P5CTLZIP, and P6CTLZIP Motifs Search Results Summary for P7CTLZIP, P8CTLZIP, and P9CTLZIP Motifs '=~~`I'11~1~~!II11.1111111 Ililllllilll I!II~'1111ii1 II(Iii=I{~{{!I-~~1~1-I{{I!{{!I{{ {I ({~{iilllll iii oi f_ -~~1iia~i_~h~~h~- I -~~ =iE !Iii 11-(III Ili!}Ililivill s P ice} oia ~1 :lal:l~ o I ioi"a_f" _ o {<i!I ! ryl~l Icl {P--!r{ IPI { ( I ~:!w~~iol._.l_ { { ( { { { (I =1 { I ! (I
{ _ al ~iaJtl_~~
~I-~~~ I~ oi~ is!$ I$ gl~~~:i:i~) ~sI~1sI o = P ^'" - =ICI 8 a!"~~1=1 '- ICI~i~{ ^I ia, i.j;~i"i~`-i=l_;o{~48!_I~ a~=' "-~I~,~ -ia 1 I~!~'~I~ i~~g~
Dl o' I. :.) o!~lo i~!~lr _-^i~l !TI^. - ^Id _~ oic to o i4 = iii I.' id 1_ a !~ Ir lc to iv to _L'-h o 0 0'. o. !.o 0 1"_ _ -~I !_ I Ie!^t'IS.=1 I^ ^!^I_ C~II I^ { I-! I:._'x'c!_ -4 I_f 1f~flslal g=!-i~l~
-t-!': I.A c .I TAT ~T o'P' - `c ~I^ ^ I^!.~ "I=~I", o I~o!P!o ~1=1a!= ~{ {~! I~-~~~lat i={ ~!~ ^I. {~I (I f~t~I I_~^

.e 1... 7!-.:. 1.. L:h~~oohh'O!o to to I: -'4 a da .. T _ ~== -- "~. ~' Iz a <-~
- y y d O6 ` N Z N Y.
L Yy J
N N y w W
wad < õZ ~Y"Z~ Z
G=_ n 0 33~< =~ < c ~ ~~S
zz Ws<4 3 = ~zr N N < z z L H w w W = N w< Ld IC 6' Z < m ~ _ ` N H J < < < BG
>>... O> Ih -C iI'1 1~1 OO< OJJJ
ySS ZON < ._.
i~,C ~_ y (... Z BCI~!~ ~ZI _I < G s m vly y Be `~: vIC H E E <Z L<Z OC S O (j N <
-1 1- O C <IN IaC c J >I d 1-1~ 3 N y IN N s>>>> C ac ((~~ D~ d d~ V ~< ~a ~ U
R Z D
d~Z U ` ~ <tvI> >~< al ~~~~ ~ O K aIBC K O BC BC OC t y ~ > N ~ N ~<fiG J ~ ~
s. L ID < ~1.+'..Ix{_fat N I^ ^ ` L S S S S N W O =_ z z w O O o zlzlZi<~ a S
Z Y Z ml<I< Z> <I Z j> >>> N O W W J BC N N BC <I< ~ BC
s! IN ~ Z ~ N1N Vd VNI !-i' N W N /-~ .. 3 a~ `~` B: ~ ~ > V ~ ~ G j 7 > > f Bc NN.. V
N a[I} Y oC ~e <I~i2IZ ~SL 2L IZ~ ~IZIZL ~ W J _ .. 1- ~ O < < < S O N < O D O
O c c w ~~~1.~.. N,l_ v ~. IN
O ~BC~ u 3 i-=JJ333w~ yBZa S S NN NNjp 0 0 N `. G OGI< 0.1 f O W .-~FIN r.3 c < < U c c c zo ` o o u <' -~ ~, c i!~ >
>!.=c B=c =I<
a O~~_{> i W_ o ZUU00w~'~ iii J acoo0 U L>SS>[iC !C N ,N N y 1-. > IF. 1~ h Z
zc. 6 = U 6G kK Z } < < >~t`yiyf1`> OVOONO O_NINNS Z <<F1-.

-x-_ <oo 9: D >>> N yT w O F J N C '-<~~S r's~ 5~515'"yg N << o 0 Z22 0~
Z~~~1~7LN [y[YL wBG ~~w . ~~1~7 O LLQUI {oi~SD~Bac =oe ~. __ 6 ai NIN N N N N
O J> C m to m m U U U U Z J J 2 L L d d i BL BC m m S 6 N N

J J J J >> s>> J
W HF W w F.~ r. F.Hh~ W H1-. f.y1..hH v1.. 1-.w JJJJOJ D J O w J J J J W wF.
y. :=HH D
N N N y<0000dOyg0 o,y 0000 NN<_q Z d Sc ~5!Si B<t ~m ~5 ~7I~~K~!~ SL ,SLe7SR~mf CK ~!m~K <<<<Z<<Z<Z<< Z< <Z<B~d~5m ~7m~ < r On < <
Td- C Bilz`, 7 7 B72 R 7 sJ7dQ C Td- d +G
1- } N N N N N N N N N N N N N N N O N N N N N
J J < < < < < J < < < J < < < < < J J pJ .J Jp J pJ of J Y
W0 BY WIW iWWIWd WIW YD.~dWWW WW WWWaCJ V 2E U62 mm1< < mlmlm mlm<mmmm<m mmm yyd~
w W W w h w I W W W ZZ y W w w y Z w w < H w w w ..~<~jj RJ ~J w w +-~ <<< Z<
<<<< y y N a NIN N N d IBC g( N N BC N N lNIN N BG 0 o a 0 < a << N N < (~ D O 0 BC

I B`{GC~~III~y ~yJyI'a<~a~aa~~yya{yGGC
w HIS Y
O g C p 1~ G J p JJ yJ J IJ pJ J e[ J J y' B( J J ..,,~I J J J J J J B~ > i cq m~ ~m J
J m~ i i > BG W B B B J
!~ s G, dl 26 D o 00 1 g i C d O G a D G 0, 0002-2 - G i=. < < f g g< C ~~~~~ c c 9? a c O
<<<< < <I<I<I<< <<I< <2<i om<mom0< o< 2<2 0<
zft !,i ! l~I~i~~~~pI~~~!~~f~i i~i~~~~sysgssds~dd'a~~d~ssssl~~~l~(~~~
Y > > > ; r > y V G m > > c wp cN = f O y > > i > > y > { D N
>
z oC !~}> m u .,zZ, z of>z >}V~1 ppp > UU D D~~==~~-An<
70 p$ < >e ^6S) O> W luww> YN NN 6.13 T T T O .9,j[ {i~
m w ._ i G d O 6 6 BG Ot N N N N N N 1~ >>> < m m m m V 6d' J J Y. B,. 6`i L
BL ~'. L =QC N M
pT'j J J J J J ) J J J J J J J J J J J J J J J J J J J J J J 0 0 0 c o o O O O C Q K
w a:

PW~

= ,.
I I~
~ I II
! P
b I

r I o a , N I
ra ' I ! P

I I I~ = N_ I ^a N S n v I n - ' n P - O
I ~p - T & 3 I P 1D n ~ -P' ti f ~ N n P O p a C - e a e a d T'~~ ~ ~= S_ 3 ss'--sd7~g =_ s~a~ra,~ ~~_3a~ N"a~~"^ Np QN

- P _--- M n- - O f a=. P. H= ~~= ~ v~_ n n n N TT" T n n > I >

ZZ ~
W K
y 9b_ ~ ~ R6~ Vl 9~ P .~. H ^ ~I~ h y y P 1~ F Y- y L' 4S ^ O Z 0 < Z o ... iC C ` C `<? Z Z Z < Z Z
Zy y LW W(y h - - v .~y.. y F_ C p. Ly' < + ` < .., Z > <
w y O } >~ W y y b N G Z W h v y y .y... -~ r ,.. P F~ -< OL p. ~` uC y y H~ y y _K S y y 1~ ~p+[ > ,- X .,, ,~.. '}-_ F- ac a[ s ~ K {WL y yam. W y > > Y ~ N
y {iW, y .y_..y.. ,y., W v ~;, _ {1_. per. v - ` ~
y > O O O ~ > O `, > y ~ > y ..y.. H > S > H } O r.. b ~ ` X ` 6 ~ N > > <~ Y
W p, ~ Y } _y 6 6 -O =~ K] y O= UQ O O y O O O ~> N Y. YC y H ~S^ > v O ~ F ` L~_ ~~= i``, A0.WL
Vl .~-. v Y i W A 0.WL
y ~y XX X Y~ > > S ZZ<< X O X O O y O y N !~ b N N y> y y 1~ F.
O g y} ~+ s X X> -> 0 0 0 t S oC ~~_ Z> O O O O y O y Vf ~C W >~ OC b' O K 6G C K oC
Tq Sq Cq. Oy _ y..100 ~ ..t u] O~ y y > X~<~' C7 C70 W ~ c =~ rj O W ~~~~ y 1- L' O h. hr. !.. a F. O y ..1 ~ O _ W u7 y O~ i W y y "! ~
d~ x yy~ y X X X X x S S
i.lyy f.1m G)y ~.~ ~ Y> <~' ~_~ NFOO"d 1nNC0 =i W O tti O W W W W y yy ~ ~~ O ~p [[ y N 0 0 0 ~ W
y~ W OUy j> ODUU a. uy F~" ( O N.i .i O..d~ W
Mw g 2 n O W S }Y
~ O O S ^ t~ti ILE
Oy~ p } C U O- .n n ^~a o_ d r OLF 8 W

L {py ~w.~1 W
__ b ~y5 ~~ L6 b.L.t .L1 a Z,~~(~C W aC _d~7g OKK Y. 1= bL ^ ~
L+ F55O^ V!~ u~~ W O~ ~"..U OO 3O pS LU2~,~~ K'.L1L Lr LZ I.
"..f ~11 ~R<ZCy m~SG007~~ V uu ,.000 2U1- ~"i1~~~+Nu+< ^~~ `oOC ~?^
= c~x f3~~a,,'".,ooo ^ pyzj 00 ZQ OI= OOOOXO pbWõOF reeV <C~ LOL ('~ ~ 4a~. a C. ~~~~Z Z
L L O O

r L L L b b b b b b L L b b L L b b b L b L L b b L L b~~~~ `~~~~ L L L L L L
=__`____ b b L b L b L L L b b 6 6 b TABLE XIV

SEARCH RESULTS SUMMARY

DEMANDES OU BREVETS VOLUMINEUX

LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET
COMPREND PLUS D'UN TOME.

NOTE: Pour les tomes additionels, veuillez contacter le Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS

THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE
THAN ONE VOLUME

THIS IS VOLUME OF _ NOTE: For additional volumes-please contact-the Canadian Patent Office

Claims (15)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. An isolated peptide having the formula:
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNAF-Z;
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLANWF-Z;
X-YTSLIHSLIEESQNQQEKNEQQLLELDKWASLWNWF-Z;
X-YTSLIHSLIEESQNQQEKNEQELLQLDKWASLWNWF-Z;
X-YTSLIHSLIEESQNQQEKNQQELLQLDKWASLWNWF-Z;
X-YTSLIHSLQEESQNQQEKNEQELLELDKWASLWNWF-Z;
X-YTSLIHSLIEQSQNQQEKNEQELLELDKWASLWNWF-Z;
X-YTSLIHSLIQESQNQQEKNEQELLELDKWASLWNWF-Z;
X-YTSLIHSLIEESQNQQEKNEQQLLELDKWASLWNWF-Z;
X-YTSLIQSLIEESQNQQEKNEQQLLELDKWASLWNWF-Z;
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLFNFF-Z;
X-YTSLIHSLIEESQNLQEKNEQELLELDKWASLWNWF-Z;
X-YTSLIHSLIEESQNQQEKLEQELLELDKWASLWNWF-Z;
X-YTSLIHSLIEESQNQQEKNEQELLEFDKWASLWNWF-Z;
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASPWNWF-Z;
X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNSF-Z;
X-LRAIEAQQHLLQLTVWQIKQLQARILAV-Z;

X-NKSLEQIWNNMTWMEWDREINNYTSLIHSLIEESQN-Z;
X-NKSLEQIWNNMTWMEWDREINNYTSLIHSLIEESQNQQEKNEQELLELDKWASLW
NWF-Z;

X-SLEQIWNNMTWMEWDREINNYTSLIHSLIEESQNQQ-Z;
X-LEQIWNNMTWMEWDREINNYTSLIHSLIEESQNQQE-Z;
X-EQIWNNMTWMEWDREINNYTSLIHSLIEESQNQQEK-Z;
X-QIWNNMTWMEWDREINNYTSLIHSLIEESQNQQEKN-Z;
X-IWNNMTWMEWDREINNYTSLIHSLIEESQNQQEKNE-Z;
X-WNNMTWMEWDREINNYTSLIHSLIEESQNQQEKNEQ-Z;
X-NNMTWMEWDREINNYTSLIHSLIEESQNQQEKNEQE-Z;
X-NMTWMEWDREINNYTSLIHSLIEESQNQQEKNEQEL-Z;
X-MTWMEWDREINNYTSLIHSLIEESQNQQEKNEQELLELDKWASLWNW-Z;

X-MTWMEWDREINNYTSLIHSLIEESQNQQEKNEQELL-Z;

X-TWMEWDREINNYTSLIHSLIEESQNQQEKNEQELLE-Z;
X-WMEWDREINNYTSLIHSLIEESQNQQEKNEQELLEL-Z;
X-WMEWDREINNYTSLIHSLIEESQNQQEKNEQELLE-Z;
X-WMEWDREINNYTSLIGSLIEESQNQQEKNEQELLE-Z;
X-MEWDREINNYTSLIHSLIEESQNQQEKNEQELLELD-Z;
X-EWDREINNYTSLIHSLIEESQNQQEKNEQELLELDK-Z;
X-WDREINNYTSLIHSLIEESQNQQEKNEQELLELDKW-Z;
X-DREINNYTSLIHSLIEESQNQQEKNEQELLELDKWA-Z;
X-REINNYTSLIHSLIEESQNQQEKNEQELLELDKWAS-Z;
X-EINNYTSLIHSLIEESQNQQEKNEQELLELDKWASL-Z;
X-INNYTSLIHSLIEESQNQQEKNEQELLELDKWASLW-Z;
X-NYTSLIHSLIEESQNQQEKNEQELLELDKWASLWNW-Z;
X-TSLIHSLIEESQNQQEKNEQELLELDKWASLWNWFN-Z;
X-SLIHSLIEESQNQQEKNEQELLELDKWASLWNWFNI-Z;
X-LIHSLIEESQNQQEKNEQELLELDKWASLWNWFNIT-Z; or X-YTSLIHSLIEESQNQQEKNEQELLELDKWASLWNWFNITNWLWLIKFI-Z;
in which X comprises an amino group, an acetyl group, a 9-fluorenylmethoxy-carbonyl group, a hydrophobic group, or a macromolecular carrier group; and Z comprises a carboxyl group, an amido group, a hydrophobic group, or a macromolecular carrier group, wherein the peptide exhibits antiviral activity.

2. The peptide of claim 1, wherein X is an amino group and Z is a carboxyl group.

3. The peptide of claim 1, wherein the hydrophobic group is a carbobenzoxyl, dansyl or t-butyloxycarbonyl hydrophobic group.

4. The peptide of claim 1, wherein X is an acetyl group or a 9-fluorenylmethoxy-carbonyl group; and Z is a t-butyloxycarbonyl hydrophobic group or an amido group.

5. The peptide of claim 4, wherein X is an acetyl group and Z is an amido group.

6. The peptide of claim 1, wherein Z is a lipid-fatty acid conjugate, a polyethylene glycol, or a carbohydrate moiety.

7. The peptide according to any one of claims 1 to 6, wherein one bond linking adjacent amino acids is a non-peptide bond, further wherein the non-peptide bond is an imino, ester, hydrazine, semicarbazide, or azo bond.

8. The peptide according to any one of claims 1 to 7, wherein at least one amino acid residue is in a D-isomer configuration.

9. The peptide according to any one of claims 1 to 8 for use as an antiviral agent in the treatment of HIV-1 infection.

10. Use of the peptide according to any one of claims 1 to 8 for the manufacture of a medicament for the treatment of HIV-1 infection.

11. Use of the peptide according to any one of claims 1 to 8 in the treatment of HIV-1 infection.

12. A pharmaceutical composition for use in inhibiting transmission of an HIV virus to a cell, comprising the peptide according to any one of claims 1 to 8 as active component and a pharmaceutically acceptable carrier.

13. An isolated nucleic acid encoding the peptide of claim 1.

14. A vector comprising a nucleic acid encoding the peptide of claim 1.

15. A peptide produced by expression of a vector comprising a nucleic acid encoding the peptide of claim 1.