Synthetic polypeptides corresponding to portions of proteinoids translated from brain-specific mRNAs, receptors, methods and diagnostics using the same

• US 4,900,811 A
• Filed: 06/03/1987
• Issued: 02/13/1990
• Est. Priority Date: 07/21/1983
• Status: Expired due to Term

• Current Assignee: The Scripps Research Institute
• Original Assignee: Scripps Clinic and Research Foundation
• Inventors: Sutcliffe, J. Gregor
• Primary Examiner(s): NOT, DEFINED
• Application Number:
  US07/058,620
• Time in Patent Office: 986 Days
• Field of Search: 435/172.3, 435/68, 530/324, 530/325, 530/326, 530/327, 530/328, 530/329, 530/330, 530/331
• US Class Current: 530/324
• 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 14/47 from mammals
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: G01N 33/567 utilising isolate of tissue or organ as binding agent
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/686 Anti-idiotype
• CPC Class Codes: G01N 33/94 involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors

Abstract

Synthetic polypeptides whose sequences correspond substantially to amino acid residue sequences of at least portions of naturally occurring proteinoids translated from brain-specific mRNAs are disclosed as are receptors, methods and diagnostics that utilize those synthetic polypeptides. The synthetic polypeptides have molecular weights less than those of their corresponding proteinoids, and induce the production of antibodies that bind to the naturally occurring proteinoid, or a derivative thereof when bound to a carrier as a conjugate and are introduced into an animal.

Claims

1. An antigenically active synthetic polypeptide constituted by at least six to about 50 amino acid residues and configured to define an epitope that is antigenically the same as determined by Western blotting assay, ELISA or immunocytochemical staining, as that of a neuroactive, mammalian brain proteinoid or a derivative thereof including the same epitope as said proteinoid, said polypeptide, when bound to a carrier as a conjugate and introduced as said conjugate into an animal, being capable of inducing the production of antibodies that bind not only to itself but also to said mammalian brain proteinoid or said derivative thereof, and said proteinoid having an amino acid residue sequence that is translated from a cytoplasmic messenger RNA present in mammalian brain cells but not in the cells of the liver, kidney, gut, lung, heart or skeletll muscle of the same species of mammal.

2. The synthetic polypeptide of claim 1 wherein said messenger RNA is polyadenylated.

3. The synthetic polypeptide of claim 1 wherein said synthetic polypeptide is neuroactive.

4. The synthetic polypeptide of claim 1 wherein said messenger RNA is present at a concentration of less than about 2 percent by weight of said cytoplasmic messenger RNA of the brain.

5. The synthetic polypeptide of claim 1 wherein said messenger RNA is present at a concentration of less than about 0.2 percent by weight of said cytoplasmic messenger RNA of the brain.

6. The synthetic polypeptide of claim 1 wherein said messenger RNA is present at a concentration of less than about 0.01 percent by weight of said cytoplasmic messenger RNA of the brain.

8. The polypeptide according to claim 1 wherein said proteinoid or said derivative thereof is present in mammalian brain cells but not in cells of the liver, kidney, gut, lung, heart or skeletal muscle of the same species of mammal.

7. A synthetic polypeptide having an amino acid residue sequence, as represented by a formula below, from left to right and in the direction from amino-terminus to carboxy-terminus, said sequence being selected from the group consisting of

(a) CIPEGLESYYTEQ;

(b) RSVSPWMSVLSEE;

(c) NVTESPSFSAGDNPHVLYSPEFRISGAPDKYESE;

(d) LLGLRGEPPELDLSYSHSDLG;

(e) PTKDSYTLTEELAEYAEIRVK;

(f) LGSERRLLGLRGEPPELDLSYSHSDLG;

(g) LLGLRGEPPELDLSYSHSDL--NH₂ ; and

(h) LGSERRLLGLRGEPPELDLSYSHSDL--NH₂.

9. A synthetic polypeptide having no more than about 50 amino acid residues and including an amino acid residue sequence, selected from the group consisting of

(a) CIPEGLESYYTEQ;

(b) RSVSPWMSVLSEE;

(c) NVTESPSFSAGDNPHVLYSPEFRISGAPDKYESE;

(d) LLGLRGEPPELDLSYSHSDLG;

(e) PTKDSYTLTEELAEYAEIRVK;

(f) LGSERRLLGLRGEPPELDLSYSHSDLG;

(g) LLGLRGEPPELDLSYSHSDL--NH₂ ; and

(h) LGSERRLLGLRGEPPELDLSYSHSDL--NH₂ ;

each of the above sequences being stated from left to right in the direction of amino-terminus to carboxy-terminus.

10. A polypeptide prepared by a method comprising the steps of:

(a) isolating messenger RNA from the brain cells of a mammal;

(b) isolating messenger RNA from another type of tissue of the same species of mammal;

(c) preparing cloned complementary DNA to said isolated messenger RN.A of step (a);

(d) hybridizing the cloned complementary DNA of step (c) with the isolated messenger RNA of step (a);

(e) hybridizing the cloned complementary DNA of step (c) with the isolated messenger RNA of step (b);

(f) determining the amount of hybridization of steps (d) and (e);

(g) selecting a clone of complementary DNA that hybridizes to the mRNA of step (a) and shows substantially no hybridization to the mRNA of step (c);

(h) determining an amino acid residue sequence of a proteinoid translated from a selected cloned complementary DNA of step (g); and

(i) synthesizing a polypeptide containing at least 6 to about 50 amino acid residues and having the same amino acid residue sequence as a portion of said proteinoid of step (h), said polypeptide, when bound to a carrier as a conjugate and introduced as said conjugate into an animal, being capable of inducing the production of antibodies that bind to itself and to said proteinoid or said derivative thereof.

11. The polypeptide in accordance with claim 10 wherein said type of tissue is selected from the group consisting of liver, kidney, gut, lung, heart and skeletal muscle.