Hepatocyte growth factor receptor antagonists and uses thereof

Details Hepatocyte growth factor receptor antagonists and uses thereof Hepatocyte growth factor receptor antagonists and uses thereof

1998-01-14

2001-04-10

Johnson, Nancy A. (Department: 1642)

Drug, bio-affecting and body treating compositions

Immunoglobulin, antiserum, antibody, or antibody fragment,...

Binds eukaryotic cell or component thereof or substance...

C424S130100, C424S138100, C424S141100, C424S143100, C424S152100, C424S155100, C424S172100, C530S387700, C530S388220, C530S388800, C530S388850, C530S389700

Reexamination Certificate

active

06214344

ABSTRACT:

FIELD OF THE INVENTION
This application relates to hepatocyte growth factor receptor antagonists. The application also relates to the use of the antagonists in therapy or diagnosis of particular pathological conditions in mammals, including cancer.
BACKGROUND OF THE INVENTION
Hepatocyte growth factor (“HGF”) functions as a growth factor for particular tissues and cell types. HGF was identified initially as a mitogen for hepatocytes [Michalopoulos et al.,
Cancer Res
., 44:4414-4419 (1984); Russel et al.,
J. Cell. Physiol
., 119:183-192 (1984); Nakamura et al.,
Biochem. Biophys. Res. Comm
., 122:1450-1459 (1984)]. Nakamura et al., supra, reported the purification of HGF from the serum of partially hepatectomized rats. Subsequently, HGF was purified from rat platelets, and its subunit structure was determined [Nakamura et al.,
Proc. Natl. Acad. Sci. USA
, 83:6489-6493 (1986); Nakamura et al.,
FEBS Letters
, 224:311-316 (1987)]. The purification of human HGF (“huHGF”) from human plasma was first described by Gohda et al.,
J. Clin. Invest
., 81:414-419 (1988).
Both rat HGF and huHGF have been molecularly cloned, including the cloning and sequencing of a naturally occurring variant lacking 5 amino acids designated “delta5 HGF” [Miyazawa et al.,
Biochem. Biophys. Res. Comm
., 163:967-973 (1989); Nakamura et al.,
Nature
, 342:440-443 (1989); Seki et al.,
Biochem. Biophys. Res. Commun
., 172:321-327 (1990); Tashiro et al.,
Proc. Natl. Acad. Sci. USA
, 87:3200-3204 (1990); Okajima et al.,
Eur. J. Biochem
., 193:375-381 (1990)].
The mature form of huHGF, corresponding to the major form purified from human serum, is a disulfide linked heterodimer derived by proteolytic cleavage of the human pro-hormone between amino acids R494 and V495. This cleavage process generates a molecule composed of an &agr;-subunit of 440 amino acids (M
r
69 kDa) and a &bgr;-subunit of 234 amino acids (M
r
34 kDa). The nucleotide sequence of the huHGF cDNA reveals that both the &agr;- and the &bgr;-chains are contained in a single open reading frame coding for a pre-pro precursor protein. In the predicted primary structure of mature huHGF, an interchain S—S bridge is formed between Cys 487 of the &agr;-chain and Cys 604 in the &bgr;-chain [see Nakamura et al.,
Nature
, supra]. The N-terminus of the &agr;-chain is preceded by 54 amino acids, starting with a methionine group. This segment includes a characteristic hydrophobic leader (signal) sequence of 31 residues and the prosequence. The &agr;-chain starts at amino acid (aa) 55, and contains four kringle domains. The kringle 1 domain extends from about aa 128 to about aa 206, the kringle 2 domain is between about aa 211 and about aa 288, the kringle 3 domain is defined as extending from about aa 303 to about aa 383, and the kringle 4 domain extends from about aa 391 to about aa 464 of the &agr;-chain.
The definition of the various kringle domains is based on their homology with kringle-like domains of other proteins (such as prothrombin and plasminogen), therefore, the above limits are only approximate. To date, the function of these kringles has not been determined. The &bgr;-chain of huHGF shows high homology to the catalytic domain of serine proteases (38% homology to the plasminogen serine protease domain). However, two of the three residues which form the catalytic triad of serine proteases are not conserved in huHGF. Therefore, despite its serine protease-like domain, huHGF appears to have no proteolytic activity, and the precise role of the &bgr;-chain remains unknown. HGF contains four putative glycosylation sites, which are located at positions 294 and 402 of the &agr;-chain and at positions 566 and 653 of the &bgr;-chain.
In a portion of cDNA isolated from human leukocytes, in-frame deletion of 15 base pairs was observed. Transient expression of the cDNA sequence in COS-1 cells revealed that the encoded HGF molecule (delta5 HGF) lacking 5 amino acids in the kringle 1 domain was fully functional [Seki et al., supra].
A naturally occurring huHGF variant has been identified which corresponds to an alternative spliced form of the huHGF transcript containing the coding sequences for the N-terminal finger and first two kringle domains of mature huHGF [Chan et al.,
Science
, 254:1382-1385 (1991); Miyazawa et al.,
Eur. J. Biochem
., 197:15-22 (1991)]. This variant, designated HGF/NK2, has been proposed to be a competitive antagonist of mature huHGF. Comparisons of the amino acid sequence of rat HGF with that of huHGF have revealed that the two sequences are highly conserved and have the same characteristic structural features. The length of the four kringle domains in rat HGF is exactly the same as in huHGF. Furthermore, the cysteine residues are located in exactly the same positions, an indication of similar three-dimensional structures [Okajima et al., supra; Tashiro et al., supra].
HGF and HGF variants are described further in U.S. Pat. Nos. 5,227,158, 5,316,921, and 5,328,837.
The HGF receptor has been identified as the product of the c-Met proto-oncogene [Bottaro et al.,
Science
, 251:802-804 (1991); Naldini et al.,
Oncogene
, 6:501-504 (1991); WO 92/13097 published Aug. 6, 1992; WO 93/15754 published Aug. 19, 1993]. The receptor is usually referred to as “c-Met” or “p190
MET
” and typically comprises, in its native form, a 190-kDa heterodimeric (a disulfide-linked 50-kDa &agr;-chain and a 145-kDa &bgr;-chain) membrane-spanning tyrosine kinase protein [Park et al.,
Proc. Natl. Acad. Sci. USA
, 84:6379-6383 (1987)]. Several truncated forms of the c-Met receptor have also been described [WO 92/20792; Prat et al.,
Mol. Cell. Biol
., 11:5954-5962 (1991)].
The binding activity of HGF to its receptor is believed to be conveyed by a functional domain located in the N-terminal portion of the HGF molecule, including the first two kringles [Matsumoto et al.,
Biochem. Biophys. Res. Commun
., 181:691-699 (1991); Hartmann et al.,
Proc. Natl. Acad. Sci
., 89:11574-11578 (1992); Lokker et al.,
EMBO J
., 11:2503-2510 (1992); Lokker and Godowski,
J. Biol. Chem
., 268:17145-117150 (1991)]. The c-Met protein becomes phosphorylated on tyrosine residues of the 145-kDa &bgr;-subunit upon HGF binding.
Certain antibodies to HGF receptor have been reported in the literature. Several such antibodies are described below.
Prat et al.,
Mol. Cell. Biol
., supra, describe several is monoclonal antibodies specific for the extracellular domain of the &bgr;-chain encoded by the c-Met gene [see also, WO 92/20792]. The monoclonal antibodies were selected following immunization of Balb/c mice with whole living GTL-16 cells (human gastric carcinoma cell line) overexpressing the Met protein. The spleen cells obtained from the immunized mice were fused with Ag8.653 myeloma cells, and hybrid supernatants were screened for binding to GTL-16 cells. Four monoclonal antibodies, referred to as DL-21, DN-30, DN-31 and DO-24, were selected.
Prat et al.,
Int. J. Cancer
, 49:323-328 (1991) describe using anti-c-Met monoclonal antibody DO-24 for detecting distribution of the c-Met protein in human normal and neoplastic tissues [see, also, Yamada et al.,
Brain Research
, 637:308-312 (1994)]. The murine monoclonal antibody DO-24 was reported to be an IgG2a isotype antibody.
Crepaldi et al.,
J. Cell Biol
., 125: 313-320 (1994) report using monoclonal antibodies DO-24 and DN-30 [described in Prat et al.,
Mol. Cell. Biol
., supra] and monoclonal antibody DQ-13 to identify subcellular distribution of HGF receptors in epithelial tissues and in MDCK cell monolayers. According to Crepaldi et al., monoclonal antibody DQ-13 was raised against a peptide corresponding to nineteen COOH-terminal amino acids (from Ser
1372
to Ser
1390
) of the human c-Met sequence.
A monoclonal antibody specific for the cytoplasmic domain of human c-Met has also been described [Bottaro et al., supra].
Several of the monoclonal antibodies referenced above are co

Affiliated with

Also associated with

Rating

Hepatocyte growth factor receptor antagonists and uses thereof does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Hepatocyte growth factor receptor antagonists and uses thereof, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Hepatocyte growth factor receptor antagonists and uses thereof will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2553616