Ligands to enhance cellular uptake of biomolecules

• US 5,994,517 A
• Filed: 11/22/1996
• Issued: 11/30/1999
• Est. Priority Date: 11/22/1995
• Status: Expired due to Fees

• Current Assignee: CELLective Dx Corporation, Paul O.P. Ts'O
• Original Assignee: Ts'O, Paul O. P.
• Inventors: Lee, Yuan C., Ts'o, Paul O. P., Hangeland, Jon J.
• Primary Examiner(s): Stucker, Jeffrey
• Application Number:
  US08/755,062
• Time in Patent Office: 1,103 Days
• Field of Search: 530/391.7, 530/391.9, 530/395, 536/23.1
• US Class Current: 530/391.9
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/6425 the peptide or protein in the drug conjugate being a receptor, e.g. CD4, a cell surface antigen, i.e. not a peptide ligand targeting the antigen, or a cell surface determinant, i.e. a part of the surface of a cell
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: C07H 15/18 Acyclic radicals, substituted by carbocyclic rings
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

Oligodeoxynucleoside methylphosphonate neoglycopeptide conjugates and related compounds for tissue specific delivery of biologically stable, nonionic oligodeoxynucleoside analogs into cells.

Claims

1. A delivery system comprising a homogeneous conjugate of formula

View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)

2. The delivery system of claim 1 wherein A is a glycosylated oligopeptide.

3. The delivery system of claim 1 wherein A is linked to L through an amide bond.

4. The delivery system of claim 1 wherein L is linked to P through a thioether bond.

5. The delivery system of claim 2 wherein A is selected from the group consisting of ##STR32## .

6. The delivery system of claim 5, wherein said ligand is YEE(ah-GalNAc)₃.

7. The delivery system of claim 5 wherein P is selected from the group consisting of deoxyriboside, riboside, and 2'"'"'-O-methylriboside.

8. The delivery system of claim 7 wherein P is an oligonucleoside with internucleotide linkages selected from the group consisting of phosphodiester, phosphorothionate diester, and methylphosphonate linkages.

9. The delivery system of claim 8, wherein said oligonucleotide contains alternating diester and methylphosphonate internucleotide linkages.

10. The delivery system of claim 1, wherein said oligonucleotide contains a radioactive nuclide.

11. The delivery system of claim 1 wherein P is ##STR33## wherein B represents A, C, G or T;

n is between 8 and 50, inclusive;

R₁ is H or a 5'"'"' conjugate linkage, R₂ and R₃ are identically H or OCH₃, and R₄ is H or a 3'"'"' conjugate linkage, with the proviso that one of R₁ and R₄ is a conjugate linkage and the other is H;

or ##STR34## wherein B represents A, C, G or T;

n is between 8 and 50, inclusive;

R₁ is H or a 5'"'"' conjugate linkage;

R₅ is H or a 3'"'"' conjugate linkage, R₂ is selected from the group consisting of O^-, CH₃, or S^-, wherein

when R₂ is O^-, R₃ is CH₃ and R₄ is O^- ;

when R₂ is CH₃, R₄ is CH₃ and R₃ may be O^- or S^- ;

when R₂ is S^-, R₄ is S^- and R₃ may be S^- or CH₃,

with the proviso that one of R₁ and R₅ is a conjugate linkage and the other is H.

12. The delivery system of claim 1, wherein said linker is heterobifunctional.

13. The delivery system of claim 1, wherein said linker is a product of a cross-linking reagent selected from the group consisting of ##STR35## and α

-citraconyl-K-(ε

-FMOC)PILFFRL.

14. A delivery system comprising a homogeous conjugate of formula A--L--P, wherein

A represents a hepatic ligand,

L represents a bifunctional linker that 1) is covalently linked to A and P in a regiospecific manner, and 2) contains a thioether linkage and an amide linkage;

and

P represents a biologically stable oligonucleotide or oligonucleotide derivative, wherein P contains internucleotide linkages resistant to enzymatic hydrolysis or biodegradation upon its release from the conjugate.

15. A delivery system for tissue specific delivery to hepatic cells, said delivery system comprising a purified conjugate that contains an oligomer and a neoglycopeptide, wherein said oligomer is oligodeoxynucleoside methylphosphonate or an analog thereof, and wherein said oligomer and said neoglycopeptide are covalently linked by a bifunctional linker.

16. The delivery system of claim 15 which is gene specific.

17. The oligodeoxynucleoside methylphosphonate neoglycopeptide conjugate [YEE(ah-GalNAc)₃ ]-SMCC-AET-U^m pT₇.