Drug-polymer conjugates coupled to a peptidic carrier

• US 7,846,893 B2
• Filed: 02/16/2006
• Issued: 12/07/2010
• Est. Priority Date: 02/16/2005
• Status: Active Grant

• Current Assignee: Rutgers University
• Original Assignee: Rutgers University
• Inventors: Zhang, Xiaoping, Wan, Li, Pooyan, Shahriar, Sinko, Patrick J., Gunaseelan, Simi, Stein, Stanley
• Primary Examiner(s): Tsang; Cecilia
• Assistant Examiner(s): GUDIBANDE, SATYANARAYAN R
• Application Number:
  US11/816,317
• Publication Number:
  US 20080188399A1
• Time in Patent Office: 1,755 Days
• Field of Search: None
• US Class Current: 514/1.2
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• 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: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents

Abstract

Monodisperse macromolecular conjugate compositions of a peptidic carrier irreversibly or reversibly conjugated with one or more effectors and one or more therapeutic agents, wherein at least one effector or therapeutic agent is attached to a pendant reactive group on said peptidic carrier via a water-soluble polymer. Monodispersity is obtained through the use of orthogonal and separate conjugation reactions.

Claims

1. A monodisperse macromolecular conjugate composition for in-vivo delivery of a therapeutic agent comprising a peptidic carrier reversibly or irreversibly conjugated with:

(a) one or more effectors and (b) one or more therapeutic agents, wherein said peptidic carrier comprises at least two different orthogonal pendant reactive groups each for specific attachment of a predetermined effector or therapeutic agent and at least one effector or therapeutic agent is attached to at least one of said pendant reactive groups on said peptidic carrier via a water-soluble polymer, and wherein said peptidic carrier comprises an amino acid sequence selected from the group consisting of;

Cys-beta Ala-Lys (SEQ ID NO;

     1);

Cys-(beta Ala-Lys)₆ (SEQ ID NO;

     2);

Acetyl-Cys-(beta Ala-beta Ala-Lys)₂-Amide (SEQ ID NO;

     3);

Acetyl-Cys-(beta Ala-(beta Ala-Lys)₄-Amide (SEQ ID NO;

     4); and

Cys-(beta Ala-beta Ala-Lys)₂ (SEQ ID NO;

     5).

2. The composition of claim 1, wherein said peptidic carrier comprises 1 to 50 amino acid subunits, amino acid-like subunits, or a combination thereof.

3. The composition of claim 2, wherein said peptidic carrier comprises 2 to 20 amino acid subunits, amino acid-like subunits, or a combination thereof.

4. The composition of claim 3, wherein said peptidic carrier comprises 3 to 8 amino acid subunits, amino acid-like subunits, or a combination thereof.

5. The composition of claim 1, wherein each pendant reactive group is independently selected from the group consisting of a primary amine, a carboxylic acid, an alcohol, a phenol, a thiol, a hydrazide, a hydrazone, a ketone, an oxime, and an aldehyde.

6. The composition of claim 1, wherein at least one therapeutic agent or effector is attached by reversible bonds to the peptidic carrier or to a water soluble polymer conjugated to said peptidic carrier.

7. The composition of claim 6, wherein said reversible bond is a disulfide or ester bond.

8. The composition of claim 5, wherein said peptidic carrier comprises one or more pendant groups formed thereon independently selected from the group consisting of a thiol, a primary amine, a carboxylic acid, an alcohol, and a phenol and said effector or therapeutic agent is conjugated thereto by means of a group reactive therewith on said effector or therapeutic agent.

9. The composition of claim 5, wherein said peptidic carrier comprises one or more pendant groups formed thereon independently selected from the group consisting of a thiol, a primary amine, a carboxylic acid, an alcohol, and a phenol and said effector or therapeutic agent is conjugated thereto by means of a water-soluble polymer on said effector or therapeutic agent.

10. The composition of claim 1 comprising 2 to 5 different orthogonal reactive groups attached to the central peptidic core.

11. The composition of claim 5, wherein there are 1 to 24 copies of each reactive group.

12. The composition of claim 11, wherein there are 3 to 8 copies of each reactive group.

13. The composition of claim 1, wherein said water-soluble polymer comprises poly(ethylene glycol).

14. The composition of claim 13, wherein said poly(ethylene glycol) is branched.

15. The composition of claim 13, wherein said poly(ethylene glycol) has a molecular weight of from 1,000 to 100,000 daltons.

16. The composition of claim 15, wherein said poly(ethylene glycol) has a molecular weight of from 2,000 to 5,000 daltons.

17. The composition of claim 5, wherein said reactive group is chemoselective with a reactive group attached to said water soluble polymer prior to conjugation.

18. The composition of claim 5, wherein said reactive group is chemoselective with a reactive group attached to said effector prior to conjugation.

19. The composition of claim 1 comprising up to 10 different effectors.

20. The composition of claim 19 comprising from 1 to 50 copies of each effector.

21. The composition of claim 20 comprising from 1 to 4 copies of each effector.

22. The composition of claim 1 comprising up to 10 different therapeutic agents.

23. The composition of claim 22 comprising from 1 to 50 copies of each therapeutic agent.

24. The composition of claim 23 comprising from 1 to 4 copies of each therapeutic agent.

25. The composition of claim 1, wherein said effector is selected from the group consisting of ligands or substrates for receptors, membrane influx or efflux transporters, and enzymes;

tissue- and cell-specific targeting groups;

permeability enhancers, and modifiers of cell-machinery, cell-signaling or cell-disposition.

26. The composition of claim 1, wherein said peptidic carrier comprises at least one first reactive group selected from the group consisting of thiols, primary amines, carboxylic acids, alcohols, phenols, hydrazides, hydrazones, ketones, oximes, and aldehydes, to which a first therapeutic agent or first effector is conjugated;

and at least one second reactive group orthogonal to said at least one first reactive group and selected from the group consisting of list thiols, primary amines, carboxylic acids, alcohols, phenols, hydrazides, hydrazones, ketones, oximes, and aldehydes to which a second therapeutic agent or second effector is conjugated.

27. The composition of claim 26, wherein said first therapeutic agent is an anti-retroviral therapeutic agent.

28. The composition of claim 26, wherein said first effector is a cell surface receptor ligand.

29. The composition of claim 28, wherein said first effector is fMLF.

30. The composition of claim 28, wherein said first effector is mannose.

31. The composition of claim 28, wherein said first effector is folate.

32. The composition of claim 26, wherein said second therapeutic agent is an antineoplastic therapeutic agent.

33. The composition of claim 32, wherein said second therapeutic agent is campothecin.

34. The composition of claim 32, wherein said second therapeutic agent is paclitaxel.

35. The composition of claim 32, wherein said second therapeutic agent is doxorubicin.

36. The composition of claim 26, wherein said second effector is a cell penetrating peptide.

37. The composition of claim 36, wherein said second effector is R.I.Tat9-KC.

38. A method for preparing a monodisperse macromolecular conjugate composition for in-vivo delivery of a therapeutic agent, wherein the macromolecular conjugate comprises a peptidic carrier conjugated with one or more effectors and one or more therapeutic agents, wherein the method comprises the steps of:

(a) providing a peptidic carrier comprising first and second orthogonal pendant reactive groups, said first orthogonal pendant reactive group for specific attachment of a predetermined effector and said second orthogonal pendant reactive group for specific attachment of a predetermined therapeutic agent, wherein said peptidic carrier is according to claim 1;

(b) attaching said effector to said peptidic carrier by means of a water-soluble polymer such that each copy of an effector is attached to the same reactive groups on each peptidic carrier in the composition; and

(c) attaching said therapeutic agent to said peptidic carrier by means of a water-soluble polymer such that each copy of a therapeutic agent is attached to the same reactive groups on each peptidic carrier in the composition.

40. A method for treating a condition linked to a viral disorder, comprising administering to a patient in need thereof an effective amount of a composition according to claim 1, wherein said therapeutic agent is an anti-infective agent having anti-viral activity and said effector aids the anti-viral action of said therapeutic agent.

41. A method for treating a condition linked to a microbial disorder, comprising administering to a patient in need thereof an effective amount of a composition according to claim 1, wherein said therapeutic agent is an anti-infective agent having anti-microbial activity and said effector aids the anti-microbial action of said therapeutic agent.

42. A method for treating a condition linked to a hyperproliferative disorder, comprising administering to a patient in need thereof an effective amount of a composition according to claim 1, wherein said therapeutic agent is an anti-neoplastic agent having anti-neoplastic activity and said effector aids the anti-neoplastic action of said therapeutic agent.

39. A monodisperse macromolecular conjugate composition comprising a peptidic carrier, wherein said peptidic carrier comprises at least two different orthogonal pendant reactive groups, wherein at least one of said pendant reactive groups on said peptidic carrier is attached to a water-soluble polymer, and wherein said peptidic carrier comprises an amino acid sequence selected from the group consisting of:

Cys-beta Ala-Lys (SEQ ID NO;

     1);

Cys-(beta Ala-Lys)₆(SEQ ID NO;

     2);

Acetyl-Cys-(beta Ala-beta Ala-Lys)₂-Amide (SEQ ID NO;

     3);

Acetyl-Cys-(beta Ala-(beta Ala-Lys)₄-Amide (SEQ ID NO;

     4); and

Cys-(beta Ala-beta Ala-Lys)₂ (SEQ ID NO;

     5).