Combination therapy with VIP antagonist

Drug – bio-affecting and body treating compositions – Radionuclide or intended radionuclide containing; adjuvant... – In an organic compound

Reexamination Certificate

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C424S001110, C424S009100, C530S324000, C514S012200

Reexamination Certificate

active

06630124

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates generally to cancer treatment. More particularly, the present invention relates to combination therapy using a polypeptide which is an antagonist of the vasoactive intestinal polypeptide (VIP) and a chemotherapeutic agent, preferably in a pharmaceutical composition.
BACKGROUND OF THE INVENTION
Vasoactive intestinal polypeptide (VIP) is a widely distributed peptide hormone which mediates a variety of physiological responses including gastrointestinal secretion, relaxation of gastrointestinal vascular and respiratory smooth muscle, lipolysis in adipocytes, pituitary hormone secretion, and excitation and hyperthermia after injection into the central nervous system. Vasoactive intestinal peptide is a 28 amino acid peptide with an amidated C-terminus, the peptide results from post transnational processing of a hormone composed of 170 amino acid residues. The VIP peptide has been shown to contain at least two functional regions, a region involved in receptor specific binding and a region involved in biological activity (Gozes and Brenneman,
Molecular Neurobiology
, 3:201-236 (1989)).
A most striking function of the 28 amino acid peptide, vasoactive intestinal peptide is the ability to promote embryonic growth (see, Gressens, P. et al.,
Nature
, 362:1558-8 (1993)). VIP is secreted by nerve endings, immune cells, and by some neoplastic cells (see, Gozes, I. et al.,
Current Medicinal Chemistry
in press (1999)). Together with its physiological actions, VIP may exert stimulating and trophic effects on neoplastic cells from neuroblastoma (see, Wollman, Y. et al.,
Brain Res
., 624:339-41 (1993)), breast (Zia, H. et al.,
Cancer Res
., 56:3486-9 (1996)) lung (see, Moody, T. W. et al,
Proc. Natl., Acad. Sci. USA
90:4345-9 (1993)) and colon cancer (see, Gozes, I. et al.,
C. Proceedings of the
15
th World Congress of Collegium Internatzonale Chirugiae
819 (1996)), inducing its own receptors by feedback mechanisms. In neuroblastoma, the most common solid malignancy in young children, VIP has been shown to have the dual effect of either inducing differentiation or stimulating cell division, depending on the cell line and the time of application. In one human neuroblastoma cell line (I), VIP produced dose-dependent stimulation of mitosis (see, Wollman, Y. et al.,
Brain Res
., 624:339-41 (1993)). In contrast, in the mouse neuroblastoma cell line Neuro2a, VIP inhibited proliferation at concentrations as low as 10
−13
M and 10
−10
M, respectively. Similarly, in lung cancer, using growth in soft agar as an index of cancer proliferation, VIP induced growth and VIP antagonists inhibited growth small and non-small cell lung cancer (see, Moody, T. W. et al.,
Proc. Natl, Acad. Sci. USA
90:4345-9 (1993), Moody, T. W. et al.,
Biomedical Res
. 1192, 13 (Suppl. 2) 131).
Gozes, et al. have developed a VIP antagonist that has proven useful for altering the function of the vasoactive intestinal peptide. (See, U.S. Pat. No. 5,217,953 issued to Gozes, et al. (1993)). This VIP antagonist was designed to retain the binding properties of VIP for its receptor, but to lack the amino acid sequence necessary for biological activity. It is believed that biological activity requires, among other factors, a phenylalanine residue at position 6. Amino acids 1-6 of native VIP were therefore replaced by a segment of neurotensin in order to alter the biological activity of native VIP and to change the membrane permeability of the peptide. Three of the six amino acids added in the neurotensin segment are basic. This is in contrast to native VIP which contains no basic residues and only one acidic residue in this region. Indeed, the concept that a tetrapeptide with basic amino acids at both ends and a proline residue adjacent to the N-terminal amino acid is essential for high activity on membrane permeability, has been proven correct for neurotensin and other peptides. As such, the VIP antagonist developed by Gozes, et al. is a hybrid molecule containing an amino acid sequence necessary for VIP receptor binding (i.e., amino acids 7-28 of VIP), and an N-terminal amino acid sequence corresponding to a portion of neurotensin.
Studies have shown that this VIP antagonist effectively antagonizes VIP-associated activity. It has been reported that this VIP antagonist inhibits the growth of VIP receptor bearing tumor cells such as, for example, lung tumor cells (i.e., NSCLC cells). (See, U.S. Pat. No. 5,217,953.)
U.S. Pat. No. 5,565,424, which issued to Gozes, et al. on Oct. 15, 1996, discloses another family of polypeptides which are antagonists of the vasoactive intestinal polypeptide. The VIP antagonists disclosed therein are 10-1000 times more efficacious, i.e., more potent in inhibiting VIP-associated activity than previous VIP antagonists. These superactive VIP antagonists were shown to inhibit cancer growth in lung and glioblastoma cells. Examples of superactive VIP antagonists include amino acid sequences referred to as the “NL-hybrid VIP antagonist”, the “S-NL-hybrid VIP antagonist” and the “S-hybrid VIP antagonist”.
Although the foregoing VIP antagonist and superactive VIP antagonists have been invaluable, there still remains a need in the art for an even more effective cancer treatment. In addition, a treatment is needed which is effective over a broader range of cancers, for solid tumors, and for more advance stages of cancer. The present invention fulfills these and other needs.
SUMMARY OF THE INVENTION
The present invention relates to a pharmaceutical composition comprising a vasoactive intestinal polypeptide (VIP) antagonist, a chemotherapeutic agent and a pharmaceutically acceptable carrier. The vasoactive intestinal polypeptide antagonists of the present invention comprise the following amino acid sequence:
R
1
-Lys-Pro-Arg-Arg-Pro-Tyr-Thr-Asp-Asn-Tyr-Thr-Arg-Leu-Arg-Lys-Gln-X
1
-Ala-X
2
-Lys-Lys-Tyr-Leu-Asn-Ser-Ile-Leu-AsnNH-R
2
.
In the above formula, R
1
and R
2
are independently selected and are functional groups including, but not limited to, the following: hydrogen, C
1
to C
20
alkyls and C
1
to C
20
acyls, provided that at least one of R
1
or R
2
is hydrogen. X
1
and X
2
, in the above formula, are independently selected from the group consisting of naturally occurring amino acids and amino acid analogs or mimetics, provided that X
1
is not methionine (SEQ ID NO:1).
Within the scope of the above formula, certain vasoactive intestinal polypeptide antagonists are preferred, namely those in which R
1
is H; R
2
is H; X
1
is a norleucine residue; and X
2
is a valine residue (hereinafter referred to as the “NL-hybrid VIP antagonist”) (SEQ ID NO:2). Equally preferred are VIP antagonists in which R
1
is CH
3
(CH
2
)
16
CO—; R
2
is H; X
1
is a norleucine residue; and X
2
is a valine residue (hereinafter referred to as the “S-NL-hybrid VIP antagonist”) (SEQ ID NO:3). Also equally preferred are VIP antagonists in which R
1
is CH
3
(CH
2
)
16
CO—; R
2
is H; X
1
is a methionine residue; and X
2
is a valine residue (hereinafter referred to as the “S-hybrid VIP antagonist”) (SEQ ID NO:4). Further equally preferred are VIP antagonists in which R
1
is a C
1
to C
20
alkyl; R
2
is H; X
1
is a norleucine residue; and X
2
is a valine residue (SEQ ID NO:5). In addition, other preferred VIP antagonists are those in which X
1
and X
2
are amino acids and amino acid analogs or mimetics of hydrophobic character.
It should be noted, however, that R
1
, R
2
, X
1
and X
2
are selected such that the VIP antagonists of the present invention have other than the following amino acid sequence:
Lys-Pro-Arg-Arg-Pro-Tyr-Thr-Asp-Asn-Tyr-Thr-Arg-Leu-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr-Leu-Asn-Ser-Ile-Leu-Asn (SEQ ID NO:6).
The pharmaceutical compositions of the present invention also comprise a chemotherapeutic agent. In certain aspects, the chemotherapeutic agents of the present invention include, but are not limited to, platinum coordination compounds, topoisomerase inhibitors, antibiotics, antimitotic alkaloids and difluoronucleosides.
In o

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