Compositions and methods for the transport of biologically active agents across cellular barriers

• US 20030161809A1
• Filed: 10/02/2001
• Published: 08/28/2003
• Est. Priority Date: 10/02/2000
• Status: Abandoned Application

• Current Assignee: AriZeke Pharmaceuticals, Inc.
• Original Assignee: AriZeke Pharmaceuticals, Inc.
• Inventors: Sheridan, Philip J., Houston, L. L., Chapin, Steven, Glynn, Jacqueline M., Hawley, Stephen B.
• Application Number:
  US09/969,748
• Publication Number:
  US 20030161809A1
• Time in Patent Office: Days
• US Class Current: 424/85.2
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/642 the peptide or protein in the drug conjugate being a cytokine, e.g. IL2, chemokine, growth factors or interferons being the inactive part of the conjugate
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4728 Calcium binding proteins, e.g. calmodulin
• CPC Class Codes: C07K 14/54 Interleukins [IL]
• CPC Class Codes: C07K 14/57527 Calcitonin gene related peptide
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 14/62 Insulins
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/4258 against anti-receptor Ig
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

Disclosed herein are complexes and compounds that pass through cellular barriers to deliver compounds into, through and out of cells, and methods of producing and using such complexes and compounds. The complexes and compounds of the invention comprise a biologically active portion and a targeting element directed to a ligand that confers transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to the ligand, with the proviso that the targeting element is not an antibody. Also disclosed are complexes and compounds that comprise two or more targeting elements directed to a ligand that confers transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to the ligand. Preferred ligands include but are not limited to the stalk of pIgR, a pIgR domain, an amino acid sequence that is conserved among pIgR'"'"'s from different animals, and one of several regions of pIgR defined herein.

Claims

1. A complex or compound comprising a biologically active portion and a targeting element directed to a ligand that confers transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to said ligand, wherein said targeting element is not an antibody.

2. The complex or compound of claim 1, wherein said targeting element is a nucleic acid.

3. The complex or compound of claim 1, wherein said ligand is the pIgR stalk or a domain, conserved sequence or region thereof.

4. The complex or compound of claim 1, wherein said ligand is a polypeptide having an amino acid sequence selected from the group consisting of LRKED, QLFVNEE, LNQLT, YWCKW, GWYWC, STLVPL, SYRTD, and KRSSK.

6. The complex or compound of claim 3, wherein said targeting element is a polypeptide derived from a calmodulin, an AP-1 Golgi adaptor or a bacterial polypeptide.

7. The complex or compound of claim 1, wherein said compound further comprises a PTD or MTS.

8. The complex or compound of claim 1, wherein said biologically active portion is a polypeptide including a peptidomimetic, a nucleic acid, a lipid, a carbohydrate, a compound or complex comprising a metal, a small molecule, or a functional derivative of any of the preceding.

9. The complex or compound of claim 1, wherein said biologically active portion is a complex or compound comprising a metal.

10. The complex or compound of claim 7, wherein said metal is selected from the group consisting of platinum(II), palladium(II), zinc and cobalt(III).

11. The complex or compound of claim 1, wherein said biologically active portion is a nucleic acid.

12. The complex or compound of claim 1, wherein said biologically active portion is a polypeptide.

13. The complex or compound of claim 12, wherein said polypeptide is selected from the group consisting of a growth factor, an interleukin, an immunogen, a hormone, an enzyme, an enzyme inhibitor, an antibody, a clotting factor, a receptor, a ligand for a receptor, a kinase, a phosphoptase, a scaffold protein, an adaptor protein, a dominant negative mutant, a protease, a signaling molecule, a regulatory molecule, transporter, a transcriptional regulator, a nucleic acid binding protein, and functional derivatives thereof.

14. The complex or compound of claim 12, wherein said polypeptide is selected from the group consisting insulin, IL-2, IL-4, hGH, sCT and hCT.

15. The complex or compound of claim 1, wherein said biologically active portion is a second targeting element that is directed to a molecular target other than said ligand.

16. The complex or compound of claim 15, wherein said complex or compound further comprises a biologically active portion that is not a targeting element.

17. The complex or compound of claim 15, wherein said second targeting element is an antibody or an antibody derivative.

22. The complex or compound of claim 1, wherein said ligand is a polypeptide having an amino acid sequence selected from the group consisting of LRKED, QLFVNEE, LNQLT, YWCKW, GWYWC, STLVPL, SYRTD, and KRSSK.

24. The complex or compound of claim 1 or claim 18, wherein said ligand is in a region of a pIgR, wherein said pIgR can be from any animal, wherein said region is selected from the group consisting of:

25. The complex or compound of claim 1 or claim 18, wherein said complex or compound, or a biologically active portion or metabolite thereof, is a cytotoxic agent, and is delivered to a cancerous or otherwise diseased cell that displays pIgR or the pIgR stalk.

37. The complex or compound of claim 1 or 18, wherein said complex or compound, or a biologically active portion or metabolite thereof, is capable of undergoing transcellular movement.

38. The complex or compound of claim 1 or 18, wherein said complex or compound, or a biologically active portion or metabolite thereof, is capable of undergoing apical to basolateral transcytosis.

39. The complex or compound of claim 1 or 18, wherein said complex or compound, or a biologically active portion or metabolite thereof, is capable of undergoing apical endocytosis.

40. The complex or compound of claim 1 or 18, wherein said complex or compound, or a biologically active portion or metabolite thereof, wherein said compound, or a biologically active portion thereof, is capable of undergoing basolateral exocytosis.

41. The complex or compound of claim 1 or 18, wherein said complex or compound, or a biologically active portion or metabolite thereof, is able to undergo intracellular transport.

42. The complex or compound of claim 1 or 18, wherein said complex or compound, or a biologically active portion or metabolite thereof, is delivered to an intracellular compartment.

43. The complex or compound of claim 1 or 18, wherein said complex or compound, or a biologically active portion or metabolite thereof, is transported across a cellular barrier.

44. A pharmaceutical composition comprising the complex or compound of claim 1 or 18.

45. The pharmaceutical composition of claim 44 further comprising one or more antiproteases or carrier polypeptides.

46. A method of delivering a biologically active agent to an animal in need thereof, comprising contacting said animal with the complex or compound of claim 1 or 18.

47. A method for transporting a biologically active agent through an epithelial or mucosal barrier, comprising contacting said epithelial or mucosal barrier with the complex or compound of claim 1 or 18.

48. A method of treating a disease in an animal, comprising contacting said animal with the complex or compound of claim 1 or 18.

49. A medical device or kit comprising the pharmaceutical composition of claim 48.

50. The compound of claim 1 or 18, wherein said complex or compound further comprises a detectable moiety.

51. A method of identifying a disease in an animal, comprising contacting said animal with the complex or compound of claim 1 or 18.

52. A diagnostic composition comprising the compound or complex of claim 1 or 18.

53. A diagnostic kit comprising the diagnostic composition of claim 52.

5. A compound comprising a biologically active portion and a targeting element directed to a ligand that confers transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to said ligand, wherein said targeting element is not an antibody, and wherein said ligand is in a region selected from the group consisting of:

18. A complex or compound comprising 2 or more targeting elements directed to one or more ligands that confer transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to said ligand.

19. The complex or compound of claim 18, wherein at least one of said targeting elements in said complex or compound is identical or substantially identical to at least one other targeting element in said compound.

20. The complex or compound of claim 18, wherein at least one of said targeting elements in said complex or compound is different from at least one other targeting element of said second compound.

21. The complex or compound of claim 18, wherein said ligand is the pIgR stalk or a domain, conserved sequence or region thereof.

23. The complex or compound of claim 18, wherein said ligand is a polypeptide having an amino acid sequence selected from the group consisting of LRKED, QLFVNEE, LNQLT, YWCKW, GWYWC, STLVPL, SYRTD, and KRSSK.

26. A compound comprising n targeting elements directed to a ligand that confers transcellular, transcytotic or paracellular transporting properties to a compound bound to said ligand, wherein one or more of desirable attributes of said compound is enhanced as compared to a second compound having m targeting elements, wherein n and m are both whole integers, and n>

m.

27. The compound of claim 26, wherein said one or more desirable attributes is a change in affinity or avidity for said ligand.

28. The compound of claim 27, wherein said pharmacological property is selected from the group consisting of half-life, decreased secretion, efficacy and selectivity.

29. A complex or compound comprising 2 or more targeting elements directed to one or more ligands that confer transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to said ligand, and at least one biologically active portion.

30. A complex or compound comprising a biologically active portion and a targeting element directed to a ligand that confers transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to said ligand, wherein said targeting element is not an antibody, wherein said complex or compound, or a biologically active portion or metabolite thereof, is absorbed from the lumen of an organ into the body of an animal.

31. A complex or compound comprising a biologically active portion and a targeting element directed to a ligand that confers transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to said ligand, wherein said targeting element is not an antibody, and at least one biologically active portion, wherein said complex or compound, or a biologically active portion or metabolite thereof, is absorbed from the lumen of an organ into the body of an animal.

33. The complex or compound of claim 31 or 32, wherein epithelial cells line the interior of said lumen.

34. The complex or compound of claim 31 or 32, wherein said lumen is selected from the group consisting of an gastrointestinal lumen, the pulmonary lumen, the nasal lumen, a nasopharyngeal lumen, a pharyngeal lumen, a buccal lumen, a sublingual lumen, a vaginal lumen, a urogenital lumen, an ocular lumen, a tympanic lumen, an ocular surface, uterine, urethral, bladder, mammary, salivary, lacrimal, respiratory sinus, biliary, sweat gland.

35. The complex or compound of claim 31 or 32, wherein said compound, or a biologically active portion or metabolite thereof, is delivered to the blood, lymph, interstitial fluid or amniotic fluid of said animal.

36. The complex or compound of claim 31, wherein said complex or compound, or a biologically active portion thereof, is delivered into the body with a pharmacokinetic profile that results in the delivery of an effective dose of said compound or a biologically active portion thereof.

32. A complex or compound comprising 2 or more targeting elements directed to one or more ligands that confer transcellular, transcytotic or paracellular transporting properties to an agent specifically bound to said ligand, and at least one biologically active portion, wherein said complex or compound, or a biologically active portion or metabolite thereof, is absorbed from the lumen of an organ into the body of an animal.