MEDICAMENTS FOR INDUCING CYTOTOXIC T-CELLS

MEDICAMENTS POUR L'INDUCTION DE CELLULES T CYTOTOXIQUES

English Abstract

The invention relates to compounds containing an amino acid with the sequence
X1-Y-X2-D-D-X3 or a nucleic acid coding this amino acid, X1 being at least one
chosen amino acid, Y being tyrosine, X2 being an amino acid chosen from the
following group: valine, isoleucine and leucine; D being aspartate and X3
being at least one other chosen amino acid, the following amino acids being
excluded: TLVLQYVDDLLL and ILVLQYVDDLLL, T being threonine, V being valine, I
being isoleucine, L being leucine and Q being glutamine. The invention also
relates to medicaments for inducing cytotoxic T-cells, containing this class
of compounds.

French Abstract

L'invention concerne des composés comprenant un acide aminé ou un acide nucléique codant pour cet acide aminé, l'acide aminé ayant la séquence ci-après: X1-Y-X2-D-D-X3, où X1 désigne au moins un acide aminé quelconque, Y désigne la tyrosine, X2 désigne un acide aminé choisi dans le groupe comprenant la valine, l'isoleucine et la leucine, D désigne un aspartate, et X3 désigne au moins un autre acide aminé quelconque, à l'exception des séquences d'acides aminés suivantes: TLVLQYVDDLLL et ILVLQYVDDLLL, où T désigne la thréonine, V la valine, I l'isoleucine, L la leucine, et Q la glutamine. L'invention concerne également des médicaments pour l'induction de cellules T cytotoxiques renfermant cette classe de composés.

Claims

8
Claims
1. A compound containing or consisting of an amino acid or
nucleic acid encoding said amino acid, wherein said
amino acid has the following sequence:
X1-Y-X2-D-D-X3, wherein
Xl - at least one of any amino acid
Y - tyrosine
X2 = one amino acid selected from the following group:
valine, isoleucine, and leucine
D = aspartate, and
X3 = at least one of any additional amino acid,
except for the following amino acid sequences:
LRVEYLDDR, TLVLQYVDDLLL, and ILVLQYVDDLLL, with
T = threonine, V = valine, I = isoleucine, L = leucine,
Q = glutamine, and R = arginine.
2. A compound according to claim 1, wherein said amino acid
sequence consists of nine amino acids.
3. A compound according to any one of the preceding claims,
wherein X1 is a sequence consisting of four or five
amino acids.
4. A compound according to any one of the preceding claims,
wherein X3 is a sequence consisting of one or two
additional amino acids.
5. A compound according to any one of the preceding claims,
wherein said amino acid sequence is a component of a
peptide.

9
6. A compouna according to any one of the preceding claims,
wherein said amino acid sequence is a component of a
protein.
7. A compound according to any one of the preceding claims,
wherein said peptide or protein is coupled to a
lipopeptide or lipoprotein, preferably to tripalmitoyl-
S-glycerylcysteinyl-seryl-serine.
8. A compound according to any one of the preceding claims,
wherein said peptide or protein is contained within a
liposome or ISCOM.
9. A compound according to any one of the preceding claims,
wherein said peptide or protein is coupled to a viral
protein.
10. A compound according to any one of the preceding claims,
wherein said peptide is selected from the following
group: HIV virus-like particles, HIV gag-particles, or
HBs antigens.
11. A compound according to any one of the preceding claims,
wherein said peptide is present as a soluble peptide-HLA
complex, preferably as a HLA-A2 tetramer.
12. A compound according to any one of the preceding claims,
wherein said peptide is present as a soluble peptide-HLA
complex bound to a liposome.
13. A compound according to any one of the preceding claims,
wherein said peptide is a component of an antigen-presenting
cell, preferably dendritic cell, macrophage,
B cell or CD4+ T cell.

10
14. A compound according to any one of the preceding claims,
wherein said amino acid sequence is selected from among
the following amino acid sequences:
IVIYQYVDDL (SEQ ID NO:1), IVICQYVDDL (SEQ ID NO:2),
IVIYQYIDDL (SEQ ID NO:3), IVICQYIDDL (SEQ ID NO:4),
ITIYQYVDDL (SEQ ID NO:5), ITICQYVDDL (SEQ ID NO:6),
ITIYQYIDDL (SEQ ID NO:7), ITICQYIDDL (SEQ ID NO:8),
IIIYQYVDDL (SEQ ID NO:9), IIICQYVDDL (SEQ ID NO:10)
IIIYQYIDDL (SEQ ID NO:11), IIICQYIDDL (SEQ ID NO:12),
MVIYQYVDDL (SEQ ID NO:13), MVICQYVDDL (SEQ ID NO:14),
MVIYQYIDDL (SEQ ID NO:15), MVICQYIDDL (SEQ ID NO:16),
VIYQYVDDL (SEQ ID NO:17), VICQYVDDL (SEQ ID NO:18),
VIYQYIDDL (SEQ ID NO:19), VICQYVDDL (SEQ ID NO:20),
LIYQYVDDL (SEQ ID NO:21), LICQYVDDL (SEQ ID NO:22),
LIYQYIDDL (SEQ ID NO:23), LICQYIDDL (SEQ ID NO:24),
TILQYVDDILL (SEQ ID NO:25), TICQYVDDILL (SEQ ID NO:26),
ILQYVDDIL (SEQ ID NO:27), ILQYIDDIL (SEQ ID NO:28),
TIVQYVDDILL (SEQ ID NO:29), TIVQYIDDILL (SEQ ID NO:30),
IVQYIDDIL (SEQ ID NO:31), IVQYIDDIL (SEQ ID NO:32),
ILVQYVDDIL (SEQ ID NO:33), ILVQYIDDIL (SEQ ID NO:34),
IIIQYVDDIL (SEQ ID NO:35), IIIQYIDDIL (SEQ ID NO:36),
ILIQYVDDIL (SEQ ID NO:37), ILIQYIDDIL (SEQ ID NO:38),
VLYQYVDDL (SEQ ID NO:39), VLCQYVDDL (SEQ ID NO:40),
VLYQYIDDL (SEQ ID NO:41), VLCQYIDDL (SEQ ID NO:42),
where C = cysteine, D = aspartate, I = isoleucine, L =
leucine, M = methionine, and Q = glutamine.
15. A compound according to any one of the preceding claims,
wherein said nucleic acid sequence is a DNA or RNA
sequence.
16. A compound according to any one of the preceding claims,
wherein said nucleic acid sequence is a component of a
plasmid or viral vector, preferably a recombinant
vaccinia virus, adenovirus, or retroviral vector.

11
17. A compound according to any one of the preceding claims,
wherein said nucleic acid sequence is a component of a
bacterial vector, preferably of a recombinant BCG or
salmonella vector.
18. A compound according to any one of the preceding claims,
wherein said nucleic acid sequence is a component of an
inactivated virus, preferably an inactivated HIV virus.
19. A medicament containing as active ingredient a compound
according to any one of the preceding claims.
20. A medicament according to claim 19 in form of a vaccine.
21. A medicament according to claim 20 containing polyvalent
vaccines.
22. A medicament according to any one of claims 19 through
21, wherein one or more cytokines are contained as
adjuvant.
23. A medicament according to any one of claims 19 through
22, wherein interleukin-2 and/or GM-CSF are contained as
adjuvant.
24. A method for the prevention or treatment of infections
with viruses, preferably mutated HIV, HIV-l, HIV-2,
HTLV-I, and HTLV-II viruses or mutated hepatitis B
viruses or a disease responding to induction of
cytotoxic T cells, consisting of administering to
patient an effective dose of a medicament comprising a
peptide of the following sequence:
X1-Y-X2-D-D-X3, wherein
X1 = at least one of any amino acid

12
Y = tyrosine,
X2 = one amino acid selected from the following group:
valine (V), isoleucine (I), and leucine (L),
D = aspartate, and
X3 = at least one additional of any amino acid,
or a nucleic acid encoding said peptide.
25. A method according to claim 24, wherein said mutant
viruses are resistant to reverse transcriptase
inhibitors.
26. A method according to claim 24 or 25, wherein said
mutant viruses are resistant to (-)-2',3'-dideoxy-3'-
thiacytidine [=3TC (lamivudine)] , (-)-(1S,4R) -4- [2-
amino-6-(cyclopropylamino)-9H-purin-9-yl]-2-
cyclopentene-1-methanol [abacavir], 2',3'-dideoxyinosine
[didanosin], 2',3'-dideoxycytidine [zalcitabin],
(-)-2'-deoxy-5-fluoro-3'-thiacytidine [=FTC].
27. A use of the following amino acid sequence:
X1-Y-X2-D-D-X3, wherein
X1 = at least one of any amino acid
Y = tyrosine (T),
X2 = one amino acid selected from the following group:
valine (V), isoleucine (I), and leucine (L),
D = aspartate, and
X3 = at least one of any additional amino acid,
or a nucleic acid encoding said peptide, for the
preparation of a medicament for the prevention or
treatment of infections with viruses, preferably mutated
HIV, HIV-1, HIV-2, HTLV-I, and HTLV-II viruses or
mutated hepatitis B viruses or a disease responding to
induction of cytotoxic T cells.

13
28. A use according to claim 27, wherein said mutant viruses
are resistant to reverse transcriptase inhibitors.
29. A use according to claim 27 or claim 28, wherein said
mutant viruses are resistant to (-)-2',3'-dideoxy-3'-
thiacytidine [=3TC (lamivudine)], (-)-(1S, 4R)-4-[2-
amino-6-(cyclopropylamino)-9H-purin-9-yl]-2-
cyclopentene-1-methanol (abacavir], 2',3'-dideoxyinosine
[didanosin], 2',3'-dideoxycytidine [zalcitabin],
(-)-2'-deoxy-5-fluoro-3'-thiacytidine [=FTC].

Description

CA 02325345 2000-09-29
Medicaments for Inducing C~totoxic T cells
The invention relates to a compound or medicament for
inducing cytotoxic T cells. The invention further relates to
a vaccine against retroviruses such as HIV-1, HIV-2, HTLV-I,
HTLV-II as well as against viruses such as hepatitis-B.
It is known from the prior art that by treatment with certain
medicaments cytotoxic T cells (CTL) may be induced.
Cytotoxic T cells eliminate inter alia virus infected cells
specifically.
In the treatment of HIV infections, inhibitors of viral
enzyme reverse transcriptase are employed. One important
reverse transcriptase inhibitor used clinically is the
medicament 3TC (which is (-)2'-deoxy-3'-thiacytidine or
lamivudine). However, HIV may become resistant to this
medicament by mutating methionine to isoleucine or valine at
codon 194 of reverse transcriptase (PNAS 1993: 90: 5653-6).
The same mutation also leads to resistance to other reverse
transcriptase inhibitors such as 1592U89 (abacavir),
zalcitabin (DDC), didanosin (DDI), and 2'deoxy-5-fluoro-
3'thiacytidine (FTC). Other viruses also have reverse
transcriptase or similar DNA polymerases that like HIV
reverse transcriptase contain the sequence YMDD at the active
catalytic center. The same mutation of methionine to valine
in the YMDD sequence of hepatitis B DNA polymerase also
induces resistance in the hepatitis B virus to 3TC, a
medicament used to fight the hepatitis B virus.
From the later published application WO 98/23755, the
following amino acid sequences are known for the treatment of
multiple sclerosis:
TLVLQYVDDLLL and ILVLQYVDDLLL, whereby T = threonine, V =
valine, I - isoleucine, L = leucine, and Q = glutamine.

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2
The object of the invention is to make available a medicament
that may be used to effectively block or positively affect
the course of viral infections, particularly HIV or hepatitis
B infections. This medicament should be suitable both for
prevention of an infection, e.g. as a preventive vaccine, and
for the therapy of an already established infection.
This object is fulfilled by the features of claim 1. Useful
embodiments of the invention result from the features of
claims 2 through 29.
According to the invention, a compound or medicament is
provided for the induction of cytotoxic T cells containing or
consisting of an amino acid or the nucleic acid encoding this
amino acid, where such amino acid has the following sequence:
X1-Y-X2-D-D-X3,
wherein X1 - at least one of any amino acid,
Y - tyrosine,
X2 - one amino acid selected from the following
group: valine, isoleucine, and leucine,
D - aspartate, and
X3 - at least one additional of any amino acid,
with the exception of the following amino acid sequences:
LRVEYLDDR, TLVLQYVDDLLL, and ILVLQYVDDLLL,
with T = threonine, V = valine, I - isoleucine, L = leucine,
Q = glutamine, and R = arginine;
as well as a method for prevention or treatment of infections
with viruses, preferably mutated HIV, HIV-1, HIV-2, HTLV-I,
or HTLV-II viruses, or mutated hepatitis-B viruses, or a
disease that responds to induction of cytotoxic T cells
consisting of administering effective doses of the medicament
comprising the amino acid with the following sequence:

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3
X1-Y-X2-D-D-X3, wherein
X1 = at least one of any amino acid,
Y - tyrosine,
X2 - one amino acid selected from the following
group: valine (V), isoleucine (I), and leucine (L),
- aspartate, and
X3 - at least one additional of any amino acid,
or nucleic acid encoding such amino acid, and/or
the use for producing a medicament comprising the following
amino acid:
X1-Y-X2-D-D-X3, wherein
X1 - at least one of any amino acid,
Y - tyrosine,
X2 - one amino acid selected from the following
group: valine, isoleucine, and leucine,
D - aspartate, and
X3 - at least one additional of any amino acid,
or a nucleic acid encoding such amino acid sequence for
production of a medicament for the prevention or treatment of
a viral infection, preferably mutated HIV, HIV-l, HIV-2,
HTLV-I, HTLV-II, or mutated hepatitis B infection, or a
disease which responds to induction of cytotoxic T cells.
The medicament in this invention induces cytotoxic T cells
which destroy cells particularly infected with mutated HIV
viruses. Surprisingly, the sequences of this invention form
T cell epitopes that are able to bind to HLA-A2 molecules and
induce specific T cell receptors against themselves. Thus,
it is possible to specifically target HIV mutants that arise
in treatment with 3TC and abacavir.

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4
According to one embodiment, the peptide consists of nine
amino acids. X1 may consist of a sequence of four or five of
any additional amino acids, X3 of one or two additional of
any amino acids. Such an amino acid sequence is especially
suited to immunize against mutant HIV viruses, but also
against other viruses, e.g. mutated hepatitis B viruses.
For purposes of immunization, the amino acid sequence may be
a component of a peptide or protein. The peptide or protein
may be coupled to a lipopeptide or lipoprotein, preferably
tripalmitoyl-S-glyceryl-cysteinyl-Beryl-serine. Depending
on the purpose, the peptide or protein may be contained
within a liposome or ISCOM (immunostimulatory complex). The
peptide or protein may however also be coupled to a viral
protein which preferably is selected from the following
group: HIV virus-like particles, HIV gag particles, or HBs
antigens.
This peptide may preferably be present as a peptide-HLA
complex in soluble form, e.g. as HLA-A2 tetramer. The above-
mentioned complex may be bound to a liposome. The peptide
may however also be part of a cell presenting an antigen,
preferably a dendritic cell, macrophage, B cell, or CD4+ T
cell. This may be achieved both by exogenic addition of the
peptide to the cell and endogenic processing of proteins
expressed in the cell.
According to the invention, this medicament may further
contain cytokines such as interleukin-2 and/or GM-CSF, or
polyvalent vaccines. It has proven especially beneficial to
select the amino acid sequence from among the following
sequences:
IVIYQYVDDL(SEQ ID N0:1), IVICQYVDDL (SEQ ID N0:2),
IVIYQYIDDL(SEQ ID N0:3), IVICQYIDDL (SEQ ID N0:4),
ITIYQYVDDL(SEQ ID N0:5), ITICQYVDDL (SEQ ID N0:6),
ITIYQYIDDL(SEQ ID N0:7), ITICQYIDDL (SEQ ID N0:8),

CA 02325345 2000-09-29
IIIYQYVDDL(SEQ ID N0:9), IIICQYVDDL (SEQ ID NO:10)
IIIYQYIDDL(SEQ ID N0:11), IIICQYIDDL (SEQ ID N0:12),
MVIYQYVDDL(SEQ ID N0:13), MVICQYVDDL (SEQ ID N0:14),
MVIYQYIDDL(SEQ ID N0:15), MVICQYIDDL (SEQ ID N0:16),
VIYQYVDDL (SEQ ID N0:17), VICQYVDDL (SEQ ID N0:18),
VIYQYIDDL (SEQ ID N0:19), VICQYIDDL (SEQ ID N0:20),
LIYQYVDDL (SEQ ID N0:21), LICQYVDDL (SEQ ID N0:22),
LIYQYIDDL (SEQ ID N0:23), LICQYIDDL (SEQ ID N0:24),
TILQYVDDILL Q ID N0:25), TICQYVDDILL
(SE (SEQ ID
N0:26),
ILQYVDDIL (SEQ ID N0:27), ILQYIDDIL (SEQ ID N0:28),
TIVQYVDDILL TIVQYIDDILL
(SEQ ID (SEQ ID
N0:29), N0:30),
IVQYIDDIL (SEQ ID N0:31), IVQYIDDIL (SEQ ID N0:32),
ILVQYVDDIL(SEQ ID N0:33), ILVQYIDDIL (SEQ ID N0:34),
IIIQYVDDIL(SEQ ID N0:35), IIIQYIDDIL (SEQ ID N0:36),
ILIQYVDDIL(SEQ ID N0:37), ILIQYIDDIL (SEQ ID N0:38),
VLYQYVDDL (SEQ ID N0:39), VLCQYVDDL (SEQ ID N0:40),
VLYQYIDDL (SEQ ID N0:41), VLCQYIDDL (SEQ ID N0:42),
where V = valine, I - isoleucine, L = leucine, M =
methionine, C = cysteine, and Q = glutamine. The nucleic
acid sequence may be a DNA or RNA sequence. It is possible
for the nucleic acid sequence to be a component of a plasmid
or viral vector, preferably of a recombinant vaccinia virus
or recombinant adenovirus, or a retroviral vector. The
nucleic acid sequence may also be a component of retroviral
vectors or attenuated retroviruses. Furthermore, the nucleic
acid may be a component of a bacterial vector, preferably a
recombinant BCG or salmonella vector or inactivated virus,
preferably HIV virus. According to the invention, the
medicament may also be used in the ex vivo production of T
cells or T cell receptors.
An additional object of the present invention is the use of a
medicament according to the present invention for prevention
or treatment of viral infection, preferably involving mutated
HIV, HIV-1, HIV-2, HTLV-I, or HTLV-II viruses, or mutated
hepatitis B viruses. The viruses may also be mutated viruses

CA 02325345 2000-09-29
6
resistant to (-)-2',3'-dideoxy-3'-thiacytidine [3TC
(lamivudine)], (-)-(1S,4R)-4-[2-amino-6-(cyclopropylamino)-
9H-purine-9-yl]-2-cyclopentene-1-methanol [abacavir], 2',3'-
dideoxyinosine [didanosin], 2',3'-dideoxycytidine
[zalcitabin], (-)-2'-deoxy-5-fluoro-3'-thiacytidine [FTC].
The efficacy of the medicament of this invention is, for
example, explained using a graphic representation of
experimental results. The following is shown here:
Fig. 1 recognition of CTL epitope VIYQYVDDL (SEQ ID N0:17)
or VIYQYIDDL (SEQ ID N0:19) by CTL clone ETMV1 and
non-recognition of CTL epitope VIYQYMDDL described
above by the same clone,
Fig. 2 specific lysis of clone of ETMV1 during titration
of peptides VIYQYVDDL (SEQ ID N0:17) and VIYQYIDDL
(SEQ ID N0:19),
Fig. 3 recognition of peptide VIYQYVDDL (SEQ ID N0:17),
and
Fig. 4 non-recognition of peptides VIYQYVDDL (SEQ ID
N0:17) and VIYQYIDDL (SEQ ID N0:19) by CTL clone
EB3, which recognizes the wild-type sequence
VIYQYMDDL.
In Fig. 1, the peptides were pre-incubated at a concentration
of 1 ~Cg/ml for one hour with autologous EBV transformed B-
cell lines labelled with 5lchromium. Four hours after
addition of clone ETMVl with an effector-target ratio of
15:1, the supernatant was harvested and the specific lysis
was calculated based on the amount of chromium released. The
p17-peptide KIRLRPGGK was used as control. As can be
inferred from Fig. 1, only peptides IVIYQYVDDL (SEQ ID NO:1),
VIYQYVDDL (SEQ ID N0:17), and VIYQYIDDL (SEQ ID N0:19) were
recognized, which contain resistance mutations against

CA 02325345 2000-09-29
7
reverse transcriptase inhibitors. Wild-type peptide
VIYQYMDDL was not recognized.
In order to achieve the results represented in Fig. 2, the
peptides described therein were pre-incubated for one hour at
concentrations as indicated with autologous EBV-transformed
B-cell lines labelled with 5lchromium. Four hours after
addition of clone ETMV1 with an effector-target ratio of 5:1,
the supernatant was harvested and the specific lysis was
calculated based on amounts of chromium released.
Fig. 3 indicates recognition of peptide VIYQYVDDL (SEQ ID
N0:17) (=RT50 M/V). It is HLA-A2 restricted. The shown
results were achieved by pre-incubating the peptide and
control peptide for one hour at a concentration of 10 ug/ml
with autologous EBV-transformed B-cell lines labelled with
5lchromium, HLA-A2-matched, or HLA-A2-negative allogenic B-
cell lines. Four hours after addition of clone ETMV1 with an
effector-target ratio of 5:1, the supernatant was harvested
and the specific lysis was calculated based on amount of
chromium released. Addition of antibodies to CD8 showed that
lysis is HLA class-I restricted.
The table shown in Fig. 4 indicates recognition of variant
peptides by clone EB3. For this purpose, peptides at
indicated concentrations were pre-incubated for one hour with
autologous EBV-transformed B-cell lines labelled with
5lchromium. Five hours after addition of clone EB3 with an
effector-target ratio of 8:1 or 10:1, the supernatant was
harvested and specific lysis was calculated based on amount
of chromium released. This clone recognizes non-mutated
wild-type sequence of HIV, but not the sequence of this
invention. It is shown that the sequence of this invention
is a new CTL epitope.

CA 02325345 2000-09-29
SEQUENCE LISTING
<110> Glaxo Group Ltd.
Harrer Dr., Thomas
<120> Medicaments for inducing cytotoxic T- cells
<130> 77673m3
<140> PCT/EP99/02249
<141> 1999-04-O1
<150> DE 19814925.5
<151> 1998-04-03
<160> 42
<170> PatentIn Vers. 2.0
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<213> Artificial Sequence
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Ile Val Ile Tyr Gln Tyr Val Asp Asp Leu
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Ile Val Ile Cys Gln Tyr Val Asp Asp Leu
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<212> PRT
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<220>
<223> Description of fArtificial Sequence: Peptide
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Ile Val Ile Tyr Gln Tyr Ile Asp Asp Leu
1 5 10

CA 02325345 2000-09-29
2
<210> 4
<211> 10
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<213> Artificial Sequence
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Ile Val Ile Cys Gln Tyr Ile Asp Asp Leu
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Ile Thr Ile Tyr Gln Tyr Val Asp Asp Leu
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Ile Thr Ile Cys Gln Tyr Val Asp Asp Leu
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Ile Thr Ile Tyr Gln Tyr Ile Asp Asp Leu
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<210> 8
<211> 10
<212> PRT
<213> Artificial Sequence
<220>

CA 02325345 2000-09-29
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<223> Description of Artificial Sequence: Peptide
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Ile Thr Ile Cys Gln Tyr Ile Asp Asp Leu
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<212> PRT
<213> Artificial Sequence
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Ile Ile Ile Tyr Gln Tyr Val Asp Asp Leu
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<213> Artificial Sequence
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Ile Ile Ile Cys Gln Tyr Val Asp Asp Leu
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<210> 11
<211> 10
<212> PRT
<213> Artificial Sequence
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<223> Description of Artificial Sequence: Peptide
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Ile Ile Ile Tyr Gln Tyr Ile Asp Asp Leu
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<210> 12
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<213> Artificial Sequence
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Ile Ile Ile Cys Gln Tyr Ile Asp Asp Leu
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CA 02325345 2000-09-29
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<211> 10
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Met Val Ile Tyr Gln Tyr Val Asp Asp Leu
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<213> Artificial Sequence
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Met Val Ile Cys Gln Tyr Val Asp Asp Leu
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<213> Artificial Sequence
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Met Val Ile Tyr Gln Tyr Ile Asp Asp Leu
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<210> 16
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 16
Met Val Ile Cys Gln Tyr Ile Asp Asp Leu
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<210> 17
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 17

CA 02325345 2000-09-29
Val Ile Tyr Gln Tyr Val Asp Asp Leu
1 5
<21D> 18
<211> 9
<212> PRT
<213> Artificial Sequence
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<223> Description of Artificial Sequence: Peptide
<400> la
Val Ile Cys Gln Tyr Val Asp Asp Leu
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<211> 9
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<213> Artificial Sequence
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Val Ile Tyr Gln Tyr Ile Asp Asp Leu
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<211> 9
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<213> Artificial Sequence
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<223> Description of Artificial Sequence: Peptide
<400> 20
Val Ile Cys Gln Tyr Ile Asp Asp Leu
1 5
<210> 21
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 21
Leu Ile Tyr Gln Tyr Val Asp Asp Leu
1 5
<21D> 22
<211> 9
<212> PRT
<213> Artificial Sequence

CA 02325345 2000-09-29
6
<220>
<223> Description of Artificial Sequence: Peptide
<400> 22
Leu Ile Cys Gln Tyr Val Asp Asp Leu
1 5
<210> 23
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 23
Leu Ile Tyr Gln Tyr Ile Asp Asp Leu
1 5
<210> 24
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 24
Leu Ile Cys Gln Tyr Ile Asp Asp Leu
1 5
<210> 25
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial 5equence:Peptide
<400> 25
Thr Ile Leu Gln Tyr Val Asp Asp Ile Leu Leu
1 5 10
<210> 26
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 26
Thr Ile Leu Gln Tyr Ile Asp Asp Ile Leu Leu
1 5 10

CA 02325345 2000-09-29
7
<210> 27
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 27
Ile Leu Gln Tyr Val Asp Asp Ile Leu
1 5
<210> 28
<211> 9
<212> P~RT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 28
Ile Leu Gln Tyr Ile Asp Asp Ile Leu
1 5
<210> 29
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 29
Thr Ile Val Gln Tyr Val Asp Asp Ile Leu Leu
1 5 10
<210> 30
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 30
Thr Ile Val Gln Tyr Ile Asp Asp Ile Leu Leu
1 5 10
<210> 31
<211> 9
<212> PRT
<213> Artificial Sequence
<220>

~
CA 02325345 2000-09-29
8
<223> Description of Artificial Sequence: Peptide
<400> 31
Ile Val Gln Tyr Ile Asp Asp Ile Leu
1 5
<210> 32
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 32
Ile Val Gln Tyr Ile Asp Asp Ile Leu
1 5
<210> 33
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 33
Ile Leu Val Gln Tyr Val Asp Asp Ile Leu
1 5 10
<210> 34
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 34
Ile Leu Val Gln Tyr Ile Asp Asp Ile Leu
1 5 10
<210> 35
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 35
Ile Ile Ile Gln Tyr Val Asp Asp Ile Leu
1 5 10
<210> 36

CA 02325345 2000-09-29
9
<211> 10
<212> PRT
<213> Artificial Sequence
<22D>
<223> Description of Artificial Sequence: Peptide
<400> 36
Ile Ile Ile Gln Tyr Ile Asp Asp Ile Leu
1 5 10
<210> 37
<211> 10
<212> PRT
<213> Artificial Sequence
<22D>
<223> Description of Artificial Sequence: Peptide
<4DD> 37
Ile Leu Ile Gln Tyr Val Asp Asp Ile Leu
1 S 10
<210> 38
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 38
Ile Leu Ile Gln Tyr Ile Asp Asp Ile Leu
1 5 10
<210> 39
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 39
Val Leu Tyr Gln Tyr Val Asp Asp Leu
1 5
<210> 40
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide

CA 02325345 2000-09-29
<400> 40
Val Leu Cys Gln Tyr Va 1 Asp Asp Leu
1 5
<210> 41
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Description of Artificial Sequence: Peptide
<400> 41
Val Leu Tyr Gln Tyr Ile Asp Asp Leu
1 5
<210> 42
<211> 9
<212> PRT
<213> Artif_cial Sequence
<220>
<223> Descr_ption of Artificial Sequence: Peptide
<400> 42
Val Leu Cys Gln Tyr Ile Asp Asp Leu
1 5