TREATMENT OF LIVER DISORDERS BY ADMINISTRATION OF RAP CONJUGATES

• US 20120251444A1
• Filed: 09/18/2007
• Published: 10/04/2012
• Est. Priority Date: 03/21/2007
• Status: Active Grant

• Current Assignee: Horizon Orphan, LLC (Amgen Inc.)
• Original Assignee: Raptor Pharmaceuticals Inc. (Amgen Inc.)
• Inventors: Starr, Christopher M., Zankel, Todd C.
• Granted Patent: US 8,795,627 B2
• Time in Patent Office: Days
• US Class Current: 424/1.69
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/704 attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin digitoxin A61K31/7048
• CPC Class Codes: A61K 31/715 Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
• CPC Class Codes: A61K 33/24 Heavy metals; Compounds thereof
• CPC Class Codes: A61K 33/243 Platinum; Compounds thereof
• CPC Class Codes: A61K 33/244 Lanthanides; Compounds thereof medicinal preparations containing radioactive lanthanides for use in therapy or testing in vivo A61K51/00
• CPC Class Codes: A61K 38/168 from plants
• CPC Class Codes: A61K 38/17 from animals; from humans enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 38/177 Receptors; Cell surface antigens; Cell surface determinants
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61N 5/1001 using radiation sources introduced into or applied onto the body; brachytherapy
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 : specific for metastasis
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 14/705 : Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention relates to the use of receptor-associate protein (RAP) and fragments and variants thereof to improve delivery of therapeutic compounds to the liver and provides methods to treat liver disorders and conditions, such as hepatic carcinoma, by administering RAP or RAP variants conjugated to active agents.

Claims

1. A method of treating a liver disorder in a subject comprising:

administering to said subject an effective amount of a conjugate comprising (a) a receptor binding moiety selected from the group consisting of Receptor Associated Protein of SEQ ID NO;

1 (RAP), RAP fragments, and RAP variants that retain RAP'"'"'s binding affinity to LRP1 of about 1-5 nM, attached to (b) an active agent for treatment of liver disorder.

2. The method of claim 1 wherein the receptor binding moiety of said conjugate is a RAP fragment or variant missing at least 200 and up to 243 amino acids from the N-terminus of SEQ ID NO:

1.

3. The method of claim 1 wherein the receptor binding moiety of said conjugate is a RAP fragment missing at least 200 and up to 243 amino acids from the N-terminus of SEQ ID NO:

1.

4. The method of any of claims 2-3 wherein said RAP fragment or variant is missing 243 amino acids from the N-terminus of SEQ ID NO:

1.

5. The method of any of claims 2-4 wherein said RAP fragment or variant is further missing at least 4 and up to 11 amino acids from the C-terminus of SEQ ID NO:

1.

6. The method of any of claims 2-4 wherein said RAP fragment or variant is further missing 11 amino acids from the C-terminus of SEQ ID NO:

1.

7. The method of claim 1 wherein said RAP fragment or variant lacks amino acids 1-143 and 320-323 of mature RAP of SEQ ID NO:

1.

8. The method of any of claims 2-7 wherein said RAP fragment or variant comprises a continuous portion of RAP d3 (SEQ ID NO:

2) that is (a) at least 71 amino acids in length and (b) comprises amino acids 256-270.

9. The method of claim 1 wherein said receptor binding moiety is a cyclic RAP peptide that is less than about 85 amino acids in length, comprising 50 contiguous amino acids that are at least 70% identical to SEQ ID NO:

4, and which binds to LRP1 with a Kd of about 1×

10<sup>−

8</sup> M or less.

10. The method of any of claims 1-9 wherein said receptor binding moiety is a RAP variant, said RAP variant comprising one or more conservative substitutions relative to native RAP of SEQ ID NO:

1.

11. The method of any one of claims 1-9 wherein said receptor binding moiety is a RAP variant, said RAP variant comprising a mutation at any one of positions 217, 249, or 251 of mature RAP.

12. The method of any one of claims 2-11 wherein said RAP variant comprises a mutation, wherein said mutation is the replacement of an acidic amino acid with a basic amino acid.

13. The method of claim 12 wherein said acidic amino acid is selected from the group consisting of D and E.

14. The method of claim 12, wherein said basic amino acid is selected from the group consisting of K and R.

15. The method to any one of claims 2-11 wherein said RAP variant comprises a mutation, wherein said mutation is the replacement of a basic amino acid with an acidic amino acid.

16. The method of claim 15, wherein said basic amino acid is selected from the group consisting of K and R.

17. The method of claim 15, wherein said acidic amino acid is selected from the group consisting of D and E.

18. The method to any one of claims 2-11 wherein said RAP variant comprises a mutation, wherein said mutation is the replacement of an amino acid selected from the group consisting of A, C, D, E, G, I, K, L, M, N, P, Q, R, S, T, and V with an amino acid selected from the group consisting of F, Y, W, and H.

19. The method of claim 1 wherein the receptor binding moiety of said conjugate is a RAP fragment or variant set out in SEQ ID NO:

9.

20. The method of any of claims 1-19, wherein the RAP, RAP fragment or RAP variant and diagnostic or therapeutic agent are linked through a linker.

21. The method of claim 20, wherein said linker is a peptide linker.

22. The method of any one of claims 1-21 wherein the receptor binding moiety is an oligomeric combination of RAP fragments or RAP variants.

23. The method of any of claims 1-22 wherein the conjugate is in a pharmaceutical composition comprising a pharmaceutically acceptable carrier, diluent or excipient.

24. The method of any of claims 1-23, wherein the liver disorder is selected from the group consisting of hepatic cancer, hepatitis, cirrhosis, fungal, rickettsial or parasitic infections, alcohol, chemical toxins, damage due to drug toxicity, metabolic liver disease, idiopathic autoimmune liver disease, biliary obstruction, hepatic steatis, cholestasis, and post-hepatectomy conditions.

25. The method of claim 24 wherein the hepatic cancer is selected from the group consisting of hepatocellular carcinoma and the active agent moiety is a cytotoxic chemotherapeutic agent.

26. The method of claim 24, wherein the disorder is a liver tumor or tumor metastases in the liver, and the therapeutic agent is a chemotherapeutic agent.

27. The method of any one of claims 1-26 wherein the active agent is a cytotoxic agent.

28. The method of claim 27 wherein the cytotoxic agent is selected from the group consisting of Mechlorethamine hydrochloride, Cyclophosphamide, Ifosfamide, Chlorambucil, Melphalan, Busulfan, Thiotepa, Carmustine, Lomustine, Dacarbazine and Streptozocin.

29. The method of any one of claims 1-26 wherein the active agent is a radioisotope.

30. The method of claim 29 wherein the radioisotope is selected from the group consisting of ¹³¹I, ¹²⁵I, ¹¹¹In, ⁹⁰Y, ⁶⁷Cu, ¹²⁷Lu, ²¹²Bi, ²¹³Bi, ²⁵⁵Fm, ¹⁴⁹Tb, ²²³Rd, ²¹³Pb, ²¹²Pb, ²¹¹At, ⁸⁹Sr, ¹⁵³Sm, ¹⁶⁶Ho, ²²⁵Ac, ¹⁸⁶Re, ⁶⁷Ga, ⁶⁸Ga and ^99mTc.

31. The method of claim 24, wherein the disorder is hepatitis caused by a virus, and the therapeutic agent is an antiviral agent.