Method of inhibiting phagocytosis

• US 5,858,981 A
• Filed: 06/07/1996
• Issued: 01/12/1999
• Est. Priority Date: 09/30/1993
• Status: Expired due to Term

• Current Assignee: University of Pennsylvania
• Original Assignee: University of Pennsylvania
• Inventors: Park, Jong-Gu, Schreiber, Alan D.
• Primary Examiner(s): Saunders, David
• Assistant Examiner(s): VANDERVEGT, FRANCOIS P
• Application Number:
  US08/657,884
• Time in Patent Office: 949 Days
• Field of Search: 514/2, 514/44, 514/18, 530/300, 530/330, 536/24.5
• US Class Current: 514/1.7
• 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: A61K 38/07 Tetrapeptides
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/70535 Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed

Abstract

The present invention relates, in general, to methods of treating diseases resulting from interactions between immune complexes and Fc receptors. In particular, the present invention relates to methods of modulating the clearance of antibody-coated cells from the circulation by inhibiting phagocytosis and to methods of modulating the interaction of immune complexes with tissue Fc receptors. Further, the invention relates to methods of modulating the activation of immunological processes mediated by Fc receptor activation resulting from antibody-antigen/receptor interaction.

Claims

1. A method of inhibiting the signal transduction of the γ

subunit of the IgE receptor Fcε

RI comprising introducing into lung cells of a mammal bearing said receptor a peptide inhibitor of Syk kinase, or a mimetic of said peptide, wherein said peptide comprises the sequence YXXL (SEQ ID NO;

8), wherein XX represents any two amino acids,

said introduction being effected under conditions such that said signal transduction is inhibited.

2. The method according to claim 1 wherein said inhibitor is said peptide.

3. The method according to claim 1 wherein XX represents the XX amino acids of a YXXL (SEQ ID NO:

8) sequence of the cytoplasmic domain of the γ

chain of Fcε

RI.

4. The method according to claim 1 wherein said lung cells are mast cells.

5. The method according to claim 1 wherein said XX represents the XX amino acids of the YXXL sequence of the cytoplasmic domain of Fcγ

RIIA.

6. The method according to claim 1 wherein said introduction is effected via aerosol administration to the lungs of a mammal.

10. The method according to claim 1 wherein said inhibitor targets the interval region between the second SH2 domain and the catalytic (kinase) domain of Syk kinase.

7. A method of inhibiting signal transduction of the γ

subunit of the IgE receptor Fcε

RI comprising introducing into cells of a mammal bearing said receptor an antisense construct that specifically binds Syk encoding sequences present in said cell under conditions such that said signal transduction is inhibited.

8. The method of claim 7 wherein said cells are present in the lungs of an asthma patient.

9. The method according to claim 8 wherein said introduction is effected by administering said construct to the lungs of said patient via aerosol or systemic administration.

13. The method according to claim 7 wherein said cells are lung cells.

14. The method according to claim 7 wherein said antisense construct is administered systemically to a mammal.

15. The method according to claim 7 wherein said antisense construct is introduced into said cells in a liposome.

11. A method of inhibiting the release of a mediator from a Syk-producing cell of a mammal comprising introducing into said cell an antisense construct that specifically binds Syk encoding sequences present in said cell.

12. The method according to claim 11 wherein said cell is present in a lung of an asthma patient.

16. The method according to claim 11 wherein said antisense construct is introduced into said cell in a liposome.

17. The method according to claim 11 wherein said cell is a lung cell present in a mammal.

18. The method according to claim 17 wherein said antisense construct is introduced into said cell via aerosol administration to the lungs of said mammal.

19. The method according to claim 11 wherein said cell is a mast cell.

20. The method according to claim 11 wherein said cell is a macrophage.

21. A method of inhibiting the phagocytic potential of a mammalian cell expressing an Fc receptor comprising introducing into said cell a construct comprising, in the 5'"'"'-3'"'"' direction of transcription:

i) a promoter functional in said cell,

ii) a segment of double-stranded DNA, the transcribed strand of which comprises a sequence complementary to endogenous mRNA encoding Syk kinase, and

iii) a termination sequence functional in said cell,

wherein said construct is introduced under conditions such that said complementary strand is transcribed and binds to said endogenous mRNA thereby reducing expression of Syk kinase and inhibiting the phagocytic and signaling potential of said cell.

22. The method according to claim 21 wherein said Fc receptor is Fcγ

RI, Fcγ

RIIA or Fcγ

RIIIA.

25. The method according to claim 21 wherein said sequence is complementary to a region of said endogenous mRNA free of secondary structure.

23. A method of inhibiting the phagocytic potential of a mammalian cell expressing an Fc receptor comprising introducing into said cell a nucleic acid complementary to an endogenous mRNA encoding Syk kinase, wherein said nucleic acid is introduced under conditions such that said nucleic acid binds to said mRNA and thereby inhibits translation of said mRNA into Syk kinase.

24. The method according to claim 23 wherein said Fc receptor is Fcγ

RI, Fcγ

RIIA or Fcγ

RIIIA.

26. The method according to claim 23 wherein said nucleic acid is complementary to a region of said mRNA free of secondary structure.

27. The method according to claim 23 wherein said nucleic acid has a stem-loop structure.

28. The method according to claim 23 wherein said nucleic acid has the sequence shown in SEQ ID NO:

21, SEQ ID NO;

22, SEQ ID NO;

23 or SEQ ID NO;

24.

29. A method of inhibiting the phagocytic potential of a Syk-producing cell comprising introducing into said cell an antisense construct that specifically binds Syk encoding sequences present in said cell under conditions such that production of Syk is inhibited.