Compositions and methods for enhancing drug delivery across and into epithelial tissues

• US 6,669,951 B2
• Filed: 02/23/2001
• Issued: 12/30/2003
• Est. Priority Date: 08/24/1999
• Status: Expired due to Fees

• Current Assignee: Kai Pharmaceuticals, Inc. (Amgen Inc.)
• Original Assignee: Cellgate, Inc. (Tbg Diagnostics Limited)
• Inventors: Wender, Paul A., McGrane, P. Leo, Rothbard, Jonathan B., Kirschberg, Thorsten A., Sista, Lalitha V. S.
• Primary Examiner(s): Russel, Jeffrey E.
• Application Number:
  US09/792,480
• Publication Number:
  US 20020127198A1
• Time in Patent Office: 1,040 Days
• Field of Search: 424/433, 424/434, 424/435, 424/436, 424/437, 424/427, 424/424, 424/449, 514/2, 514/11, 514/12, 514/13, 514/14, 514/15, 514/16, 514/17, 514/169, 514/634, 514/635, 514/636, 530/300, 530/317, 530/321, 530/324, 530/325, 530/326, 530/327, 530/328, 530/329, 530/330, 564/225, 564/230, 564/233, 564/236, 564/243
• US Class Current: 424/436
• CPC Class Codes: A61K 31/155 Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
• CPC Class Codes: A61K 31/337 having four-membered rings, e.g. taxol
• CPC Class Codes: A61K 31/436 the heterocyclic ring system containing a six-membered ring having oxygen as a ring hetero atom, e.g. rapamycin
• CPC Class Codes: A61K 31/496 Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
• CPC Class Codes: A61K 31/573 substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
• 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 38/13 Cyclosporins
• CPC Class Codes: A61K 47/54 the modifying agent being an organic compound
• CPC Class Codes: A61K 47/55 the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
• CPC Class Codes: A61K 47/555 pre-targeting systems involving an organic compound, other than a peptide, protein or antibody, for targeting specific cells
• CPC Class Codes: A61K 47/557 the modifying agent being biotin
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• 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: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61K 47/665 the pre-targeting system, clearing therapy or rescue therapy involving biotin-(strept) avidin systems
• CPC Class Codes: A61K 49/0043 Fluorescein, used in vivo
• CPC Class Codes: A61K 49/0056 Peptides, proteins, polyamino acids
• CPC Class Codes: A61K 49/085 conjugated systems
• CPC Class Codes: A61K 49/146 the peptide being a polyamino acid, e.g. poly-lysine
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/10 : Laxatives
• CPC Class Codes: A61P 21/00 : Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 23/00 : Anaesthetics
• CPC Class Codes: A61P 25/02 : for peripheral neuropathies
• CPC Class Codes: A61P 27/02 : Ophthalmic agents
• CPC Class Codes: A61P 27/06 : Antiglaucoma agents or miotics
• CPC Class Codes: A61P 27/12 : for cataracts
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• 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 33/02 : Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 : Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/38 : of the suprarenal hormones
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: B82Y 5/00 : Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 7/06 : having 5 to 11 amino acids
• CPC Class Codes: C07K 7/64 : Cyclic peptides containing only normal peptide links

Abstract

This invention provides compositions and methods for enhancing delivery of drugs and other agents across epithelial tissues, including the skin, gastrointestinal tract, pulmonary epithelium, ocular tissues and the like. The compositions and methods are also useful for delivery across endothelial tissues, including the blood brain barrier. The compositions and methods employ a delivery enhancing transporter that has sufficient guanidino or amidino sidechain moieties to enhance delivery of a compound conjugated to the reagent across one or more layers of the tissue, compared to the non-conjugated compound. The delivery-enhancing polymers include, for example, poly-arginine molecules that are preferably between about 6 and 25 residues in length.

Claims

1. A method of targeting a compound to a gastrointestinal epithelium of an animal, the method comprising administering to the gastrointestinal epithelium a conjugate comprising the compound and a delivery-enhancing transporter,

wherein:

i. the delivery-enhancing transporter comprises fewer than 50 subunits and comprises at least 5 guanidino or amidino moieties; and

ii. the conjugate has a structure selected from the group consisting of structures 3, 4, 5, and 6 as follows;

wherein;

R¹ comprises the compound;

X is a linkage formed between a functional group on the compound and a terminal functional group on the linking moiety;

Y is a linkage formed from a functional group on the transport moiety and a functional group on the linking moiety;

Ar is an aryl group having the attached radicals arranged in an ortho or para configuration, which aryl group can be substituted or unsubstituted;

A is N or CH;

R² is hydrogen, alkyl, aryl, acyl, or allyl;

R³ comprises the delivery-enhancing transporter;

R⁴ is S, O, NR⁶ or CR⁷R⁸;

R⁵ is H, OH, SH or NHR⁶;

R⁶ is hydrogen, alkyl, aryl, acyl or allyl;

R⁷ is hydrogen or an alkyl;

R⁸ is hydrogen or an alkyl;

k and m are each independently selected from 1 and 2; and

n is 1 to 10,

thereby increasing delivery of the conjugate into the gastrointestinal epithelium compared to delivery of the compound in the absence of the delivery-enhancing transporter.

2. The method of claim 1, wherein delivery of the conjugate into the gastrointestinal epithelium is increased at least two-fold compared to delivery of the compound in the absence of the delivery-enhancing transporter.

3. The method of claim 1, wherein delivery of the conjugate into the gastrointestinal epithelium is increased at least ten-fold compared to delivery of the compound in the absence of the delivery-enhancing transporter.

4. The method of claim 1, wherein the linker is a releasable linker.

5. The method of claim 1, wherein the subunits are amino acids.

6. The method of claim 1, wherein X is selected from the group consisting of —

C(O)O—

, —

C(O)NH—

, —

OC(O)NH—

, —

S—

S—

, —

C(S)O—

, —

C(S)NH—

, —

NHC(O)NR—

, —

SO₂NH—

, —

SONH—

, phosphate, phosphonate, phosphinate, and CR⁷R⁸, wherein R⁷ and R⁸ are each independently selected from the group consisting of H and alkyl.

7. The method of claim 1, wherein the conjugate comprises structure 3, Y is NH, and R² is methyl, ethyl, propyl, butyl, allyl, benzyl or phenyl.

8. The method of claim 1, wherein the conjugate comprises structure 3, R² is benzyl;

k, m, and n are each 1, and X is —

OC(O)—

.

9. The method of claim 1, wherein the conjugate comprises structure 4;

R⁴ is S;

R⁵ is NHR⁶; and

R⁶ is hydrogen, methyl, allyl, butyl or phenyl.

10. The method of claim 1, wherein the conjugate comprises structure 4;

R⁵ is NHR⁶;

R⁶ is hydrogen, methyl, allyl, butyl or phenyl; and

k and m are each 1.

11. The method of claim 1, wherein the conjugate comprises structure 6, R⁴ is S;

R⁵ is NHR⁶; and

R⁶ is hydrogen, methyl, allyl, butyl or phenyl.

12. The method of claim 1, wherein the conjugate comprises at least two delivery-enhancing transporters.

13. The method of claim 1, wherein the conjugate is administered orally.

14. The method of claim 1, wherein the conjugate is administered as a suppository.

15. The method of claim 1, wherein the delivery-enhancing transporter comprises a non-peptide backbone.

16. The method of claim 1, wherein the delivery-enhancing transporter is not attached to an amino acid sequence to which the delivery enhancing transporter molecule is attached in a naturally occurring protein.

17. The method of claim 1, wherein the delivery-enhancing transporter comprises from 5 to 25 guanidino or amidino moieties.

18. The method of claim 17, wherein the delivery-enhancing transporter comprises between 7 and 15 guanidino moieties.

19. The method of claim 17, wherein the delivery-enhancing transporter comprises at least 6 contiguous guanidino and/or amidino moieties.

20. The method of claim 1, wherein the delivery-enhancing transporter consists essentially of 5 to 50 amino acids, at least 50 percent of which amino acids are arginines or analogs thereof.

21. The method of claim 20, wherein the delivery-enhancing transporter comprises 5 to 25 arginine residues or analogs thereof.

22. The method of claim 21, wherein at least one arginine is a D-arginine.

23. The method of claim 22, wherein all of the arginines are D-arginines.

24. The method of claim 20, wherein at least 70 percent of the amino acids that comprise the delivery-enhancing transporter are arginines or arginine analogs.

25. The method of claim 20, wherein the delivery-enhancing transporter is seven contiguous D-arginines.

26. The method of claim 1, wherein the compound is a therapeutic for the disease selected from the group consisting of inflammatory bowel disease, colon cancer, ulcerative colitis, gastrointestinal ulcers, constipation and imbalance of salt and water absorption.

27. The method of claim 1, wherein the compound is selected from the group consisting of immunosuppressiVes, ascomycins, corticosteroids, laxatives, antibiotics and anti-neoplastic agents.

28. The method of claim 1, wherein the compound is targeted to the iliem and/or colon.

34. The method of claim 4 wherein the conjugate has a structure selected from the group consisting of structures 3, 4, and 5, as follows:

wherein;

R¹ comprises the compound;

X is a linkage formed between a functional group on the compound and a terminal functional group on the linking moiety;

Y is a linkage formed from a functional group on the transport moiety and a functional group on the linking moiety;

A is N or CH;

R² is hydrogen, alkyl, aryl, acyl, or allyl;

R³ comprises the delivery-enhancing transporter;

R⁴ is S, O, NR⁶ or CR⁷R⁸;

R⁵ is H, OH, SH or NHR⁶;

R⁶ is hydrogen, alkyl, aryl, acyl or allyl;

R⁷ and R⁸ are independently selected from hydrogen or alkyl;

k and m are each independently selected from 1 and 2; and

n is 1 to 10.

35. The method of claim 34, wherein X is selected from the group consisting of —

C(O)O—

, —

C(O)NH—

, —

OC(O)NH—

, —

S—

S—

, —

C(S)O—

, —

C(S)NH—

, —

NHC(O)NH—

, —

SO₂NH—

, —

SONH—

, phosphate, phosphonate, phosphinate, and CR⁷R⁸, wherein R⁷ and R⁸ are each independently selected from the group consisting of H and alkyl.

36. The method of claim 34, wherein the conjugate comprises structure 3, Y is NH, and R² is methyl, ethyl, propyl, butyl, allyl, benzyl or phenyl.

37. The method of claim 34, wherein R² is benzyl;

k, m, and n are each 1, and X is —

OC(O)—

.

38. The method of claim 34, wherein the conjugate comprises structure 4;

R⁴ is S;

R⁵ is NHR⁶; and

R⁶ is hydrogen, methyl, allyl, butyl or phenyl.

39. The method of claim 34, wherein the conjugate comprises structure 4;

R⁵ is NHR⁶;

R⁶ is hydrogen, methyl, allyl, butyl or phenyl; and

k and m are each 1.

29. A method for enhancing delivery of a compound into and across one or more layers of an animal ocular epithelial tissue, the method comprising:

administering to the ocular epithelial tissue a conjugate comprising the compound and a delivery-enhancing transporter,

wherein;

i. the compound is attached to the delivery-enhancing transporter through a linker, and

ii. the delivery-enhancing transporter comprises fewer than 50 subunits and comprises at least 5 guanidino or amidino moieties, thereby increasing delivery of the conjugate into and across the ocular epithelial tissue compared to delivery of the compound in the absence of the delivery-enhancing transporter.

30. The method of claim 29, wherein delivery of the conjugate into the ocular epithelial tissue is increased at least two-fold compared to delivery of the compound in the absence of the delivery-enhancing transporter.

31. The method of claim 29, wherein delivery of the conjugate into the ocular epithelial tissue is increased at least ten-fold compared to delivery of the compound in the absence of the delivery-enhancing transporter.

32. The method of claim 29, wherein the linker is a releasable linker.

33. The method of claim 29, wherein the subunits are amino acids.

40. The method of claim 29, wherein the conjugate comprises structure 6 as follows:

wherein;

R¹ the compound;

X is a linkage formed between a functional group on the compound and a terminal functional group on the linking moiety;

Y is a linkage formed from a functional group on the transport moiety and a functional group on the linking moiety;

Ar is an aryl group having the attached radicals arranged in an ortho or para configuration, which aryl group can be substituted or unsubstituted;

R³ comprises the delivery-enhancing transporter;

R⁴ is S, O, NR⁶ or CR⁷R⁸;

R⁵ is H, OH, SH or NHR⁶;

R⁶ is hydrogen, alkyl, aryl, arylalkyl, acyl or allyl;

R⁷ and R⁸ are independently selected from hydrogen or alkyl; and

k and m are each independently selected from 1 and 2.

41. The method of claim 40, wherein X is selected from the group consisting of —

C(O)O—

, —

C(O)NH—

, —

OC(O)NH—

, —

S—

S—

, —

C(S)O—

, —

C(S)NH—

, —

NHC(O)NH—

, —

SO₂NH—

, —

SONH—

, phosphate, phosphonate, phosphinate, and CR⁷R⁸, wherein R⁷ and R⁸ are each independently selected from the group consisting of H and alkyl.

42. The method of claim 40, wherein R⁴ is S;

R⁵ is NHR⁶; and

R⁶ is hydrogen, methyl, allyl, butyl or phenyl.

43. The method of claim 29, wherein the conjugate comprises at least two delivery-enhancing transporters.

44. The method of claim 29, wherein the conjugate is administered as an eye drop.

45. The method of claim 29, wherein the conjugate is administered as an injection.

46. The method of claim 29, wherein the delivery-enhancing transporter comprises a non-peptide backbone.

47. The method of claim 29, wherein the delivery-enhancing transporter is not attached to an amino acid sequence to which the delivery enhancing transporter molecule is attached in a naturally occurring protein.

48. The method of claim 29, wherein the delivery-enhancing transporter comprises from 5 to 25 guanidino or amidino moieties.

49. The method of claim 48, wherein the delivery-enhancing transporter comprises between 7 and 15 guanidino moieties.

50. The method of claim 48, wherein the delivery-enhancing transporter comprises at least 6 contiguous guanidino and/or amidino moieties.

51. The method of claim 29, wherein the delivery-enhancing transporter consists essentially of 5 to 50 amino acids, at least 50 percent of which amino acids are arginines or analogs thereof.

52. The method of claim 51, wherein the delivery-enhancing transporter comprises 5 to 25 arginine residues or analogs thereof.

55. The method of claim 51, wherein at least 70 percent of the amino acids that comprise the delivery-enhancing transporter are arginines or arginine analogs.

56. The method of claim 51, wherein the delivery-enhancing transporter is seven contiguous D-arginines.

57. The method of claim 29, wherein the compound is a therapeutic for the disease selected from the group consisting of conjunctivitis, bacterial infections, viral infections, dry eye, and glaucoma.

58. The method of claim 29, wherein the compound is selected from the group consisting of antibacterial compounds, antiviral compounds, cyclosporin, ascomycins, and corticosteroids.

88. The method of claim 29, wherein the conjugate is transported into an ocular tissue selected from the group consisting of cornea, iris, lens, vitreus, vitreus humor, aqueous humor, sclera, and optic nerve.

53. The method of claim wherein at least one arginine is a D-arginine.

54. The method of claim 53, wherein all of the arginines are D-arginines.

59. A method of targeting a compound to a gastrointestinal epithelium of an animal, the method comprising administering to the gastrointestinal epithelium a conjugate comprising the compound and a delivery-enhancing transporter,

wherein:

i. the compound is attached to the delivery-enhancing transporter through a linker;

ii. the compound is a therapeutic for the disease selected from the group consisting of inflammatory bowel disease, colon cancer, ulcerative colitis, gastrointestinal ulcers, constipation and imbalance of salt and water absorption; and

ii. the delivery-enhancing transporter comprises fewer than 50 subunits and comprises at least 5 guanidino or amidino moieties, thereby increasing delivery of the conjugate into the gastrointestinal epithelium compared to delivery of the compound in the absence of the delivery-enhancing transporter.

60. The method of claim 59, wherein the compound is therapeutic for inflammatory bowel disease.

61. The method of claim 59, wherein the compound is therapeutic for colon cancer.

62. The method of claim 59, wherein the compound is therapeutic for ulcerative colitis.

63. The method of claim 59, wherein the compound is therapeutic for gastrointestinal ulcers.

64. The method of claim 59, wherein the compound is therapeutic for constipation.

65. The method of claim 59, wherein the compound is therapeutic for an imbalance of salt and water absorption.

66. A method for enhancing delivery of a compound into and across one or more layers of an animal epithelial tissue, the method comprising buccally administering a conjugate comprising the compound and a delivery-enhancing transporter, wherein:

i. the compound is attached to the delivery-enhancing transporter through a linker; and

ii. the delivery-enhancing transporter comprises fewer than 50 subunits and comprises at least 5 guanidino or amidino moieties, thereby increasing delivery of the conjugate into and across one or more layers of the animal epithelial tissue compared to delivery of the compound in the absence of the delivery-enhancing transporter.

67. The method of claim 66, wherein the linker is a releasable linker.

68. The method of claim 66, wherein the subunits are amino acids.

69. The method of claim 66, wherein the conjugate has a structure selected from the group consisting of structures 3, 4, 5 and 6, as follows:

wherein;

R¹ comprises the compound;

X is a linkage formed between a functional group on the compound and a terminal functional group on the linking moiety;

Y is a linkage formed from a functional group on the transport moiety and a functional group on the linking moiety;

Ar is an aryl group having the attached radicals arranged in an ortho or para configuration, which aryl group can be substituted or unsubstituted;

A is N or CH;

R² is hydrogen, alkyl, aryl, acyl, or allyl;

R³ comprises the delivery-enhancing transporter;

R⁴ is S, O, NR⁶ or CR⁷R⁸;

R⁵ is H, OH, SH or NHR⁶;

R⁶ is hydrogen, alkyl, aryl, acyl or allyl;

R⁷ is hydrogen or an alkyl;

R⁸ is hydrogen or an alkyl;

k and m are each independently selected from 1 and 2; and

n is 1 to 10.

70. The method of claim 69, wherein X is selected from the group consisting of —

C(O)O—

, —

C(O)NH—

, —

OC(O)NH—

, —

S—

S—

, —

C(S)O—

, —

C(S)NH—

, —

NHC(O)NH—

, —

SO₂NH—

, —

SONH—

, phosphate, phosphonate, phosphinate, and CR⁷R⁸, wherein R⁷ and R⁸ are each independently selected from the group consisting of H and alkyl.

71. The method of claim 69, wherein the conjugate comprises structure 3, Y is NH, and R² is methyl, ethyl, propyl, butyl, allyl, benzyl or phenyl.

72. The method of claim 69, wherein R² is benzyl;

k, m, and n are each 1, and X is —

OC(O)—

.

73. The method of claim 69, wherein the conjugate comprises structure 4;

R⁴ is S;

R⁵ is NHR6; and

R⁶ is hydrogen, methyl, allyl, butyl or phenyl.

74. The method of claim 69, wherein the conjugate comprises structure 4;

R⁵ is NHR⁶;

R⁶ is hydrogen, methyl, allyl, butyl or phenyl; and

k and m are each 1.

75. The method of claim 69, wherein the conjugate comprises structure 6, R⁴ is S;

R⁵ is NHR⁶; and

R⁶ is hydrogen, methyl, allyl, butyl or phenyl.

76. The method of claim 66, wherein the conjugate comprises at least two delivery-enhancing transporters.

77. The method of claim 69, wherein the delivery-enhancing transporter comprises a non-peptide backbone.

78. The method of claim 66, wherein the delivery-enhancing transporter is not attached to an amino acid sequence to which the delivery enhancing transporter molecule is attached in a naturally occurring protein.

79. The method of claim 66, wherein the delivery-enhancing transporter comprises from 5 to 25 guanidino or amidino moieties.

80. The method of claim 66, wherein the delivery-enhancing transporter comprises between 7 and 15 guanidino moieties.

81. The method of claim 66, wherein the delivery-enhancing transporter comprises at least 6 contiguous guanidino and/or amidino moieties.

82. The method of claim 66, wherein the delivery-enhancing transporter consists essentially of 5 to 50 amino acids, at least 50 percent of which amino acids are arginines or analogs thereof.

83. The method of claim 66, wherein the delivery-enhancing transporter comprises 5 to 25 arginine residues or analogs thereof.

84. The method of claim 83, wherein at least one arginine is a D-arginine.

85. The method of claim 84, wherein all of the arginines are D-arginines.

86. The method of claim 82, wherein at least 70 percent of the amino acids that comprise the delivery-enhancing transporter are arginines or arginine analogs.

87. The method of claim 66, wherein the delivery-enhancing transporter is seven contiguous D-arginines.