Protein-polycation conjugates

• US 5,354,844 A
• Filed: 03/09/1990
• Issued: 10/11/1994
• Est. Priority Date: 03/16/1989
• Status: Expired due to Term

• Current Assignee: Boehringer Ingelheim International GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Original Assignee: Boehringer Ingelheim International GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Inventors: Beug, Hartmut, Cotten, Matthew, Wagner, Ernst, Birnstiel, Max L.
• Primary Examiner(s): Russel, Jeffrey E.
• Application Number:
  US07/492,460
• Time in Patent Office: 1,677 Days
• Field of Search: 530/358, 530/400, 530/402, 530/395, 530/345, 530/350, 530/358, 530/330, 530/331, 536/27, 536/23.1, 935/2, 935/3, 935/6, 935/52, 514/8, 514/21, 514/44
• US Class Current: 530/345
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 15/64 General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

The invention relates to a system for transporting nucleic acids into the cell, which is effected by receptor-mediated endocytosis. Using a transferrin-polycation conjugate, a complex can be formed with the polyanionic nucleic acid. This complex is bound to the transfertin receptor, which is highly regulated in growing cells, and absorbed into the cell. Suitable nucleic acids include those which inhibit specific genes or the RNA function, such as antisense oligonucleotides or ribozymes or the genes coding for them. The invention further relates to a process for introducing nucleic acids into the cells, transferrin-polycation/nucleic acid complexes and pharmaceutical preparations containing them.

Claims

1. A transferrin-polycation conjugate which is capable of forming a complex with a nucleic acid coding for a protein, wherein said conjugate when complexed with said nucleic acid, is capable of being internalized into erythroblasts by endocytosis mediated by the transferrin receptor, wherein said protein is capable of being expressed after said complex is internalized into erythroblasts, wherein the molar ratio of transferrin to polycation is 10:

1 to 1;

4, and wherein said polycation is a protamine or a homologous polypeptide comprising positively charged amino acids.

2. The conjugate of claim 1, wherein said polycation is a polylysine.

3. The conjugate of claim 1, wherein said polycation is protamine.

4. The conjugate of claim 1, wherein said polycation contains between about 20 and 500 positive charges.