Vectors encoding brain derivable polypeptide factors

• US 5,011,777 A
• Filed: 06/29/1990
• Issued: 04/30/1991
• Est. Priority Date: 01/25/1985
• Status: Expired due to Fees

• Current Assignee: Oncogen
• Original Assignee: Oncogen
• Inventors: Marquardt, Hans, Todaro, George J., Shoyab, Mohammed
• Primary Examiner(s): Schwartz, Richard A.
• Assistant Examiner(s): Ulm, John D.
• Application Number:
  US07/546,248
• Time in Patent Office: 305 Days
• Field of Search: 435/69.1, 435/172.3, 436/547, 530/350, 530/387
• US Class Current: 435/320.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/4703 Inhibitors; Suppressors
• CPC Class Codes: C07K 16/18 against material from animals or humans

Abstract

Novel polypeptides, polynucleotide sequences, DNA constructs and compositions are provided for the preparation and use of polypeptides associated with naturally occurring polypeptides found in brains. The low molecular weight polypeptides either are growth inhibitors for neoplastic cells without inhibiting normal cells or affect GABA-ergic transmission. The polypeptides find use in inhibiting neoplastic growth, modulating diazepam receptor response, and detecting receptors for the polypeptides. Antibodies are provided in conjunction with the polypeptides, which may be used together or separately for detecting the presence of the polypeptides.

Claims

1. A DNA expression construct comprising a DNA sequence encoding a first polypeptide sequence having at least one epitopic site cross reactive with a second polypeptide found in mammalian brain capable of specifically binding to the diazepam receptor in vitro, said first polypeptide including substantially the following sequence or a physiologically active fragment of at least about 30 amino acids thereof:

##STR6## where plurality of amino acids at a site indicates that any amino acid at that site may be substituted by any other amino acid at that site and an asterisk (*) intends a bond may be employed at that site, said DNA sequence joined in proper orientation to at least one of a promoter other than the wild-type promoter, a terminator other than the wild-type terminator, or a replication system other than the wild-type replication system.

2. A DNA expression construct according to claim 1, wherein said replication system, promoter and terminator are functional in a eukaryotic host.

3. A DNA expression construct comprising a DNA sequence encoding an active polypeptide, said polypeptide characterized by eluting in the 20% to 26% n-propanol fraction in aqueous 0.1M trifluoroacetic acid-n-propanol, being of from about 8 to 18 kDal, inhibiting tumor cell or T-cell growth and being found in mammalian brains, or a physiologically active fragment thereof, said DNA sequence joined in proper orientation to at least one of a promoter other than the wild-type promoter, a terminator other than the wild-type terminator, or a replication system other than the wild-type replication system.

4. A DNA expression construct according to claim 3, wherein said replication system, promoter and terminator are functional in a prokaryotic host.

5. A DNA expression construct comprising a DNA sequence encoding a polypeptide having substantially the same amino acid composition as membrane protein as set forth in FIG. 1 or a fragment thereof, said fragment beginning at methionine 37, 68 or 69, said DNA sequence joined in proper orientation to at least one of a promoter other than the wild-type promoter, a terminator other than the wild-type terminator, or a replication system other than the wild-type replication system.

6. A DNA expression construct according to claim 5, wherein said replication system, promoter and terminator are functional in a prokaryotic host.