Claims
- 1. An isolated peptide which mimics the carbohydrate epitope GlcAβ1→3Galβ1→4GlcNAc or or sulfate -3GlcAβ1→3Galβ1→4GlcNAc.
- 2. An isolated peptide comprising an amino acid sequence X1 X2 X3 X4 X5 L/V X6 X7 X8 X9 X10 X11 X12 X13 X14, wherein each residue can be independently selected as follows (SEQ ID NO: 1):
X1 is T, S, A or P; X2 is L, I, V, M, F, H, W or N; X3 is T, S, A, H, Y, F, W, N, D or E; X4 is R, Q, K, T, S or A; X5 is V, I, L, M, R, Q or K; X6 is T, S, A, Y, F, H, W, N, L, I, V or M; X7 is D, E, V, L, I, M, F, Y, K W or N; X8 is V, I, L, M, S, A, T, R, Q or K; X9 is Y, F, H, W, D, E, I, V, L, M or N; X10 is R, Q, K, W, Y, F, H, N, V, I, L, M or G; X11 is G, Y, F, H, W, N, S, A, T, I, V, L, M; X12 is R, Q, K, H, N, Y, F, W, I, V, L or M; X13 is L, V, I, M, T, S or A; and X14 is S, T, A, P, G, R, Q or K; and variants, analogs and active fragments thereof.
- 3. An isolated peptide comprising an amino acid sequence F L H T R L X1 X2 X3 X4 X5 X6 X7 X8 X9, wherein each residue can be independently selected as follows (SEQ ID NO: 2):
X1 is T, S, A, Y, F, H, W, N, L, I, V or M; X2 is D, E, V, L, I, M, F, Y, H, W or N; X3 is V, I, L, M, S, A, T, R, Q or K; X4 is Y, F, H, W, D, E, I, V, L, M or N; X5 is R, Q, K, W, Y, F, H, N, V, I, L, M or G; X6 is G, Y, F, H, W, N, S, A, T, I, V, L, M; X7 is R, Q, K, H, N, Y, F, W, I, V, L or M; X8 is L, V, I, M, T, S or A; and X9 is S, T, A, P, G, R, Q or K; and variants, analogs and active fragments thereof.
- 4. An isolated peptide comprising an amino acid sequence F L H T R L F V X1 X2 X3 X4 X5 X6 X7, wherein each residue can be independently selected as follows (SEQ ID NO: 3):
X1 is V, I, L, M, S, A, T, R, Q or K; X2 is Y, F, H, W, D, E, I, V, L, M or N; X3 is R, Q, K, W, Y, F, H, N, V, I, L, M or G; X4 is G, Y, F, H, W, N, S, A, T, I, V, L, M; X5 is R, Q, K, H, N, Y, F, W, I, V, L or M; X6 is L, V, I, M, T, S or A; and X7 is S, T, A, P, G, R, Q or K; and variants, analogs and active fragments thereof.
- 5. An isolated peptide comprising the amino acid sequence F L H T R L F V S D W Y H T (SEQ ID NO: 7).
- 6. An isolated peptide comprising the amino acid sequence F L H T R L F V (SEQ ID NO: 8).
- 7. An isolated peptide comprising the amino acid sequence TRLFR(V/F) (SEQ ID NO: 39).
- 8. An isolated peptide comprising the amino acid sequence TRLF(R)V (SEQ ID NO: 40).
- 9. An isolated peptide comprising the amino acid sequence TRLF (SEQ ID NO: 41).
- 10. An isolated peptide having the amino acid sequence set out in any of SEQ ID NO: 27-38.
- 11. A method for promoting neural growth and/or remyelination and/or neuroprotection in vivo in the central nervous system of a mammal comprising administering to said mammal a neural growth and/or remyelination and/or neuroprotection promoting amount of the peptide of claim 1, which molecule is capable of overcoming inhibitory molecular cues found on glial cells and myelin and promoting said neural growth, active fragments thereof, cognates thereof, congeners thereof, mimics thereof, antagonists thereof, antibodies thereto, analogs thereof, secreting cells thereof and soluble molecules thereof.
- 12. The method of claim 11 further comprising administering to said mammal a neural growth and/or remyelination and/or neuroprotection promoting amount of a neural cell adhesion molecule.
- 13. The method of claim 12 wherein said neural cell adhesion molecule is selected from the group consisting of L1, N-CAM and myelin-associated glycoprotein, laminin, fibronectin, N-cadherin, BSP-2/D2 (mouse N-CAM), 224-1A6-A1, L1-CAM, NILE (rat L1), Nr-CAM, TAG-1 (axonin-1), Ng-CAM and F3/F11/contactin.
- 14. A method for promoting neural growth and/or remyelination and/or neuroprotection in vivo in the central nervous system of a mammal comprising administering to said mammal a neural growth promoting amount of an agent, said agent comprising a neural cell adhesion molecule, which molecule is capable of overcoming inhibitory molecular cues found on glial cells and myelin and promoting said neural growth, active fragments thereof, secreting cells thereof and soluble molecules thereof, said agent being modified by recombinant or chemical means to have the peptide of any of claim 1 attached thereto.
- 15. The method of claim 14 wherein said neural cell adhesion molecule is selected from the group consisting of L1, N-CAM and myelin-associated glycoprotein, laminin, fibronectin, N-cadherin, BSP-2/D2 (mouse N-CAM), 224-1A6-A1, L1-CAM, NILE (rat L1), Nr-CAM, TAG-1 (axonin-1), Ng-CAM and F3/F11/contactin.
- 16. A method for enhancing memory, comprising administering to the brain of a mammal in need of such enhancement, an amount of the peptide of claim 1 effective to enhance the memory of the mammal.
- 17. A method of claim 16 which further comprises administering to the brain of said mammal an amount of a neural cell adhesion molecule effective to enhance the memory of the mammal.
- 18. A method for enhancing memory, comprising delivering to the cells of the brain of a mammal in need of such enhancement, a vector which allows for the expression of the peptide of any of claim 1.
- 19. The method for enhancing memory in accordance with any of claims 12 or 14, which comprises a method for inhibiting the onset or progression, or treating the presence or consequences of Alzheimers disease or dementia in a mammal.
- 20. A method for increasing synaptic efficacy in the CNS of a mammal comprising administering to the brain of the mammal, an amount of the peptide of claim 1 effective to increase synaptic efficacy in the brain of the mammal.
- 21. The method of claim 20, wherein the increase in synaptic efficacy is demonstrated by the stabilization of long term potentiation.
- 22. A method of promoting neuroprotection and/or neuronal survival in a mammal comprising delivering to the cells of the brain of a mammal in need thereof, a vector which allows for the expression of the peptide of claim 1.
- 23. The method of claim 22 which comprises a method for inhibiting the development or onset, or treating the presence in a mammal of a condition selected from the group consisting of apoptosis, necrosis, Alzheimers disease, dementia, Parkinsons disease, multiple sclerosis, acute spinal cord injury, chronic spinal cord injury, any of the foregoing where neurodegeneration occurs or may occur, and combinations thereof.
- 24. A method for inhibiting axonal cell death and enhancing myelination and remyelination in the central nervous system of a mammal comprising administering to said mammal a therapeutically effective amount of a peptide of claim 1, which peptide is capable of overcoming inhibitory molecular cues found on glial cells and myelin and promoting said neural growth, active fragments thereof, cognates thereof, congeners thereof, mimics thereof, antagonists thereof, antibodies thereto, analogs thereof, secreting cells thereof and soluble molecules thereof.
- 25. A pharmaceutical composition for the modulation of neural growth in the central nervous system of a mammal, comprising a therapeutically effective amount of a peptide of claim 1, which peptide is capable of overcoming inhibitory molecular cues found on glial cells and myelin and promoting said neural growth, variants, analogs, active fragments thereof, and secreting or expressing cells thereof, and a pharmaceutically acceptable carrier.
- 26. A derivative of the peptide of claim 1, capable of mimicking the carbohydrate epitope GlcAβ1→3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc, having one or more chemical moieties attached thereto.
- 27. The derivative of claim 26, wherein at least one of said chemical moieties is a water-soluble polymer capable of enhancing solubility of said peptide.
- 28. The derivative of claim 26, wherein at least one of said chemical moeities is a molecule which facilitates transfer or transport across the blood brain barrier.
- 29. The derivative of claim 28, wherein said molecule is selected from the group consisting of a biocompatible hydrophobic molecule, transferrin, ApoE or ApoJ.
- 30. The derivative of claim 26, wherein at least one of said chemical moieties is a molecule having multiple sites for peptide attachment and capable of binding at least two of said peptides simultaneously to generate a multimeric peptide structure.
- 31. The derivative of claim 30 where said molecule is selected from the group of BSA, ovalbumin, human serum albumin, polyacrylamide, beads and synthetic fibers (biodegradable and non-biodegradable).
- 32. The derivative of claim 26, wherein at least one of said chemical moieties is a neural cell adhesion molecule.
- 33. The derivative of claim 26, wherein at least one of said chemical moieties is a branched or unbranched polymer.
- 34. The derivative of claim 26, wherein at least one of said chemical moieties is N-terminally attached to said peptide.
- 35. The derivative of claim 26, wherein at least one of said chemical moieties is C-terminally attached to said peptide.
- 36. A DNA sequence which encodes a peptide of claim 1.
- 37. A DNA sequence which encodes a peptide of claim 1, or a fragment thereof, selected from the group consisting of:
(A) DNA capable of encoding the peptide set out in any of SEQ ID NOS: 1-8 and 27-41; (B) DNA sequences that hybridize to any of the foregoing DNA sequences under standard hybridization conditions; and (C) DNA sequences that code on expression for an amino acid sequence encoded by any of the foregoing DNA sequences.
- 38. A recombinant DNA molecule comprising a DNA sequence or degenerate variant thereof and a heterologous nucleotide sequence, wherein said DNA sequence or degenerate variant encodes a peptide of claim 1, or a fragment thereof, selected from the group consisting of:
(A) DNA capable of encoding the peptide set out in any of SEQ ID NOS: 1-8 and 27-41; (B) DNA sequences that hybridize to any of the foregoing DNA sequences under standard hybridization conditions; and (C) DNA sequences that code on expression for an amino acid sequence encoded by any of the foregoing DNA sequences.
- 39. The recombinant DNA molecule of claim 38, wherein said DNA sequence is operatively linked to an expression control sequence.
- 40. The recombinant DNA molecule of claim 38, wherein said expression control sequence is selected from the group consisting of the early or late promoters of SV40 or adenovirus, the lac system, the trp system, the TAC system, the TRC system, the major operator and promoter regions of phage λ, the control regions of fd coat protein, the promoter for 3-phosphoglycerate kinase, the promoters of acid phosphatase and the promoters of the yeast α-mating factors the promoters of neural cell adhesion molecules, the promoter of L1, the gFAP promoter, and the promoter for myelin basic protein.
- 41. A unicellular host transformed with a recombinant DNA molecule comprising a DNA sequence or degenerate variant thereof, which encodes a peptide of claim 1, or a fragment thereof, selected from the group consisting of:
(A) DNA capable of encoding the peptide set out in any of SEQ ID NOS: 1-8 and 27-41; (B) DNA sequences that hybridize to any of the foregoing DNA sequences under standard hybridization conditions; and (C) DNA sequences that code on expression for an amino acid sequence encoded by any of the foregoing DNA sequences; wherein said DNA sequence is operatively linked to an expression control sequence.
- 42. The unicellular host of claim 41 wherein the unicellular host is selected from the group consisting of E. coli, Pseudomonas, Bacillus, Streptomyces, yeasts, CHO, R1.1, B-W, L-M, COS 1, COS 7, BSC1, BSC40, and BMT10 cells, plant cells, insect cells, mammalian cells, human cells and neural cells in tissue culture.
- 43. A cloning vector which comprises the DNA sequence according to claim 36 and a heterologous nucleotide sequence.
- 44. An expression vector which comprises the DNA sequence according to claim 36 and a heterologous nucleotide sequence.
- 45. The expression vector of claim 44 wherein the heterologous nucleotide sequence is an expression control sequence.
- 46. The expression vector of claim 44 wherein the heterologous nucleotide sequence encodes a neural cell adhesion molecule.
- 47. A method for detecting the presence or activity of a peptide or compound, said peptide or compound capable of mimicking the carbohydrate epitope GlcAβ1→3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc wherein said peptide or compound is measured by:
A. contacting a sample in which the presence or activity of said peptide or compound is suspected with a binding partner of said peptide or compound under conditions that allow binding of said peptide or compound to said binding partner to occur; and B. detecting whether binding has occurred between said peptide or compound from said sample and the binding partner; wherein the detection of binding indicates that presence or activity of said peptide or compound in said sample.
- 48. The method of claim 47 wherein the binding partner is selected from the group consisting of an antibody which recognizes GlcAβ1→Galβ1→4GlcNAc; an antibody which recognizes sulfate -3GlcAβ1→3Galβ1→4GlcNAc; L2-412 antibody; HNK-1antibody; a polypeptide molecule which binds or otherwise interacts with GlcAβ1→3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc; laminin; P-selectin; L-selectin; and a neural cell adhesion molecule.
- 49. A method of testing the ability of a drug or other entity to mimic the carbohydrate epitope GlcAβ1→3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc which comprises:
a. adding CNS neurons to a cell culture system; b. adding the drug or other entity under test to the cell culture system; c. measuring the neuronal outgrowth of the CNS neurons; and d. correlating a difference in the level of neuronal outgrowth of cells in the presence of the drug relative to a control culture to which no drug is added to the ability of the drug to mimic the carbohydrate epitope GlcAβ1 →3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc.
- 50. A test kit for the demonstration of a molecule capable of binding GlcAβ1→3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc in a eukaryotic cellular sample, comprising:
A. a predetermined amount of a detectably labeled compound or peptide, said peptide or compound capable of mimicking the carbohydrate epitope GlcAβ1→3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc; B. other reagents; and C. directions for use of said kit.
- 51. A test kit for demonstrating the presence of a molecule capable of binding GlcAβ1→3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc in a eukaryotic cellular sample, comprising:
A. a predetermined amount of a compound or peptide, said peptide or compound capable of mimicking the carbohydrate epitope GlcAβ1→3Galβ1→4GlcNAc or sulfate -3GlcAβ1→3Galβ1→4GlcNAc; B. a predetermined amount of a specific binding partner of said compound or peptide; C. other reagents; and D. directions for use of said kit; wherein either said compound or peptide or said specific binding partner are detectably labeled.
- 52. A pharmaceutical composition for promoting neural growth and/or remyelination and/or neuroprotection, comprising a therapeutically effective amount of the peptide of claim 1 or variants or analogs thereof and a pharmaceutically acceptable carrier.
- 53. The pharmaceutical composition of claim 52 further comprising a therapeutically effective amount of a neural cell adhesion molecule.
- 54. A method for preventing, ameliorating or blocking viral infection of a mammal comprising administering to said mammal an effective amount of the peptide of claim 1, variants thereof, analogs thereof, active fragments thereof or derivatives thereof.
- 55. The method of claim 54 wherein the viral infection is the result of the human immunodeficiency virus.
- 56. A method for preventing, ameliorating or blocking neuropathy in a mammal comprising administering to said mammal an effective amount of the peptide of claim 1, variants thereof, analogs thereof, active fragments thereof or derivatives thereof, wherein said neuropathy is viral-mediated, immune-mediated or the result of trauma.
- 57. A pharmaceutical composition for preventing, ameliorating or blocking viral infection comprising a therapeutically effective amount of the peptide of claim 1 or variants, analogs, derivatives or active fragments thereof and a pharmaceutically acceptable carrier.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a continuation-in-part of copending application Serial No. 60/121,327 filed Feb. 24, 1999, of copending application Serial No. 60/155,492 filed Sep. 23, 1999, of which the instant application claims the benefit of the filing date pursuant to 35 U.S.C. §119, and which is incorporated herein by reference in its entirety.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60121327 |
Feb 1999 |
US |
|
60155492 |
Sep 1999 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09511956 |
Feb 2000 |
US |
Child |
10186867 |
Jul 2002 |
US |