Glutamate transport modulatory compounds and methods

• US 20060121488A1
• Filed: 02/25/2004
• Published: 06/08/2006
• Est. Priority Date: 02/26/2003
• Status: Abandoned Application

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: Rothstein, Jeffrey D
• Application Number:
  US10/546,860
• Publication Number:
  US 20060121488A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C12Q 1/6897 involving reporter genes operably linked to promoters
• CPC Class Codes: G01N 2333/70571 for neuromediators, e.g. serotonin receptor, dopamine receptor
• CPC Class Codes: G01N 2800/2871 Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event
• CPC Class Codes: G01N 33/9406 Neurotransmitters

Abstract

Described herein are methods of modulating excitatory amino acid transporter (EAAT) protein expression, methods of treating disease and disease symptoms, methods of identifying compounds that modulate EAAT protein expression, and compounds useful for modulating EAAT protein expression and treating disease and disease symptoms.

Claims

1. A method of increasing EAAT2 protein expression comprising the step of contacting a cell with at least one EAAT2 expression promoting agent.

2. The method of claim 1, wherein the EAAT2 expression promoting agent is a compound identified by a screening assay comprising the steps of:

a) contacting a test compound with a nucleic acid molecule comprising a cDNA molecule and having a nucleotide sequence which is at least about 60% identical to the nucleotide sequence of SEQ ID NO;

1, 2, 3, or 4, wherein the nucleic acid molecule is capable of directing mRNA expression from a promoterless reporter vector, or a complement thereof, or a cell comprising said nucleic acid molecule; and

b) determining whether expression of the mRNA or the polypeptide encoded by the cDNA is modulated,

thereby identifying a compound which is an EAAT2 expression promoting agent.

3. The method of either claim 1 wherein EAAT2 protein expression is increased in vivo.

4. The method of either claim 1 wherein EAAT2 protein expression is increased in vitro.

5. The method of claim 1, wherein the EAAT2 expression promoting agent is an antibiotic, an anti-hypertensive, a neurotransmitter, an antibacterial, an anti-inflammatory, a steroid derivative, or an anti-septic.

6. The method of claim 1, wherein the EAAT2 expression promoting agent comprises at least one structural element selected from heterocycles comprising at least one ring sulfur atom, tertiary amines, quaternary ammonium salts, steroids, polyols, polyketide, guanidine, urea, or arsenate.

7. The method of claim 6, wherein the EAAT2 expression promoting agent comprises at least one structural element selected from tertiary amines, quaternary ammonium salts, polyketides, steroidal ring systems and heterocycles having one or two rings, at least one sulfur ring atom and 0, 1, or 2 nitrogen ring atoms.

8. The method of claim 1, wherein the EAAT2 expression promoting agent increases EAAT2 production by 200% or more relative to non-regulated EAAT2 production.

9. The method of claim 8, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, vancomycin hydrochloride, thiaphenicol, ceftriaxone, ceftriaxone sodium, 1,3-dipropyl-8-cyclopentylxanthine, cephapirin sodium, actinospectacin, cefoperazole, cefoperazole sodium, acivicin, acetylcholine, chloramphenicol, vidarabine, atenolol, oxytetracycline, glafenine, oxymetazoline hydrochloride, gallamine, perillic acid (−

), amitriptyline hydrochloride, tetracaine hydrochloride, disopyramide phosphate, sisomicin sulfate, ketamine hydrochloride, xylazine, bicuculline, flurbiprofen, cefadroxil, bacampicillin hydrochloride, tiapride hydrochloride, norethindrone acetate, bergaptene, carisoprodol, citiolone, piroxicam, erythromycin ethylsuccinate, furegrelate sodium, albendazole, dihydrostreptomycin sulfate, aloin, fenoprofen, flutamide, ampicillin, ampicillin sodium, amprolium, sparteine sulfate, medroxyprogesterone acetate, alexidine hydrochloride, clindamycin hydrochloride, cephalothin, cephalothin sodium, daidzein, meclizine hydrochloride, lindane, bromopride, N-(3-trifluoromethylphenyl)piperazine hydrochloride, enoxolone, ipratropium bromide, bufexamac, gluconolactone, rifampin, hydroxychloroquine, coleoforsin, chloroxine, oxidopamine hydrochloride, camptothecin, nafcillin sodium, mianserin hydrochloride, acetarsol, prilocaine hydrochloride, deferoxamine mesylate, hexamethonium bromide, methenamine, paraxanthine, harmalol hydrochloride, pyrithione zinc, hydrocortisone butyrate, acetazolamide, aminoglutethimide, meclofenoxate hydrochloride, 2-phenpropylamino-5-nitrobenzoic acid, amiodarone hydrochloride, aconitine, hydroxyprogesterone caproate, and diosmin.

10. The method of claim 1, wherein the EAAT2 expression promoting agent increases EAAT2 production by 300% or more relative to non-regulated EAAT2 production

11. The method of claim 10, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, vancomycin hydrochloride, thiaphenicol, ceftriaxone, ceftriaxone sodium, 1,3-dipropyl-8-cyclopentylxanthine, cephapirin sodium, actinospectacin, cefoperazole sodium, acivicin, acetylcholine, chloramphenicol, vidarabine, atenolol, oxytetracycline, glafenine, and oxymetazoline hydrochloride.

12. The method of claim 1, wherein the EAAT2 expression promoting agent increases EAAT2 production by 400% or more relative to non-regulated EAAT2 production.

13. The method of claim 12, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, and vancomycin hydrochloride.

14. The method of claim 1, wherein the EAAT2 expression promoting agent increases EAAT2 production by 600% or more relative to non-regulated EAAT2 production.

15. The method of claim 14, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, and quinapril.

16. A method for decreasing extracellular glutamate concentration in a mammal, the method comprising the step of administering at least one EAAT2 expression promoting agent to the mammal.

17. The method of claim 16, wherein the mammal has been identified as in need of such treatment.

18. The method of claim 16, wherein the EAAT2 expression promoting agent is a compound identified by a screening assay comprising the steps of:

a) contacting a test compound with a nucleic acid molecule comprising a cDNA molecule and having a nucleotide sequence which is at least about 60% identical to the nucleotide sequence of SEQ ID NO;

1, 2, 3, or 4, wherein the nucleic acid molecule is capable of directing mRNA expression from a promoterless reporter vector, or a complement thereof, or a cell comprising said nucleic acid molecule; and

b) determining whether expression of the mRNA or the polypeptide encoded by the cDNA is modulated,

thereby identifying a compound which is an EAAT2 expression promoting agent.

19. The method of claim 16, wherein EAAT2 protein expression is increased in vivo.

20. The method of claim 16, wherein EAAT2 protein expression is increased in vitro.

21. The method of claim 16, wherein the EAAT2 expression promoting agent is an antibiotic, an anti-hypertensive, a neurotransmitter, an antibacterial, an anti-inflammatory, a steroid derivative, or an anti-septic.

22. The method of claim 16, wherein the EAAT2 expression promoting agent comprises at least one structural element selected from heterocycles comprising at least one ring sulfur atom, tertiary amines, quaternary ammonium salts, steroids, polyols, polyketide, guanidine, urea, or arsenate.

23. The method of claim 22, wherein the EAAT2 expression promoting agent comprises at least one structural element selected from tertiary amines, quaternary ammonium salts, polyketide, steroidal ring systems and heterocycles having one or two rings, at least one sulfur ring atom and 0, 1, or 2 nitrogen ring atoms.

24. The method of claim 16, wherein the EAAT2 expression promoting agent increases EAAT2 production by 200% or more relative to non-regulated EAAT2 production.

25. The method of claim 24, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, vancomycin hydrochloride, thiaphenicol, ceftriaxone, ceftriaxone sodium, 1,3-dipropyl-8-cyclopentylxanthine, cephapirin sodium, actinospectacin, cefoperazole, cefoperazole sodium, acivicin, acetylcholine, chloramphenicol, vidarabine, atenolol, oxytetracycline, glafenine, oxymetazoline hydrochloride, gallamine, perillic acid (−

), amitriptyline hydrochloride, tetracaine hydrochloride, disopyramide phosphate, sisomicin sulfate, ketamine hydrochloride, xylazine, bicuculline, flurbiprofen, cefadroxil, bacampicillin hydrochloride, tiapride hydrochloride, norethindrone acetate, bergaptene, carisoprodol, citiolone, piroxicam, erythromycin ethylsuccinate, furegrelate sodium, albendazole, dihydrostreptomycin sulfate, aloin, fenoprofen, flutamide, ampicillin, ampicillin sodium, amprolium, sparteine sulfate, medroxyprogesterone acetate, alexidine hydrochloride, clindamycin hydrochloride, cephalothin, cephalothin sodium, daidzein, meclizine hydrochloride, lindane, bromopride, N-(3-trifluoromethylphenyl)piperazine hydrochloride, enoxolone, ipratropium bromide, bufexamac, gluconolactone, rifampin, hydroxychloroquine, coleoforsin, chloroxine, oxidopamine hydrochloride, camptothecin, nafcillin sodium, mianserin hydrochloride, acetarsol, prilocaine hydrochloride, deferoxamine mesylate, hexamethonium bromide, methenamine, paraxanthine, harmalol hydrochloride, pyrithione zinc, hydrocortisone butyrate, acetazolamide, aminoglutethimide, meclofenoxate hydrochloride, 2-phenpropylamino-5-nitrobenzoic acid, amiodarone hydrochloride, aconitine, hydroxyprogesterone caproate, and diosmin.

26. The method of claim 16, wherein the EAAT2 expression promoting agent increases EAAT2 production by 300% or more relative to non-regulated EAAT2 production.

27. The method of claim 25, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, vancomycin hydrochloride, thiaphenicol, ceftriaxone, ceftriaxone sodium, 1,3-dipropyl-8-cyclopentylxanthine, cephapirin sodium, actinospectacin, cefoperazole sodium, acivicin, acetylcholine, chloramphenicol, vidarabine, atenolol, oxytetracycline, glafenine, and oxymetazoline hydrochloride.

28. The method of claim 16, wherein the EAAT2 expression promoting agent increases EAAT2 production by 400% or more relative to non-regulated EAAT2 production.

29. The method of claim 28, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, and vancomycin hydrochloride.

30. The method of claim 16, wherein the EAAT2 expression promoting agent increases EAAT2 production by 600% or more relative to non-regulated EAAT2 production.

31. The method of claim 30, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, and quinapril.

32. The method of claim 16, wherein the mammal is a primate.

33. The method of claim 32, wherein the mammal is a human.

34. The method of claim 16, wherein the extracellular glutamate concentration is reduced by at least about 50% relative to non-regulated extracellular glutamate concentration.

35. The method of claim 16, wherein the extracellular glutamate concentration is reduced by at least about 75% relative non-regulated concentration.

55. The method of claim 35, wherein the extracellular glutamate concentration is reduced by at least about 75% relative non-regulated concentration.

63. The method of claim 16, wherein the EAAT2 expression promoting agent is a compound identified by a screening assay comprising the steps of

a) contacting a cell that expresses EAAT2 with a test compound;

and

b) determining whether expression of the EAAT2 in the cell is modulated in the presence of the test compound compared to in the absence of the test compound, thereby identifying a compound which modulates expression of the EAAT2.

64. The method of claim 16, wherein the EAAT2 expression promoting agent is a β

-lactam antibiotic.

65. The method of claim 16, wherein the EAAT2 expression promoting agent is a penicillin class, cephalosporin class, carbapenam class or monobactam class compound.

36. A method of treating a mammal suffering from or susceptible to a disease or disorder associated with altered glutamate transmission, the method comprising the step of administering to the mammal a therapeutic amount of at least one EAAT2 expression promoting agent capable of increasing EAAT2 expression.

37. The method of claim 36, wherein the mammal has been identified as in need of such treatment.

38. The method of claim 36, wherein the disease or disorder associated with altered glutamate transmission is a neurological disease.

39. The method of claim 38, wherein the neurological disease is selected from the group consisting of Parkinson'"'"'s disease, Huntington'"'"'s disease, Alzheimer'"'"'s disease, multiple sclerosis, amyotrophic lateral sclerosis, acute neurological diseases, epilepsy, spinal cord injury, brain trauma, glaucoma, and psychiatric disorders.

40. The method of claim 36, wherein the EAAT2 expression promoting agent is a compound identified by a screening assay comprising the steps of

a) contacting a test compound with a nucleic acid molecule comprising a cDNA molecule and having a nucleotide sequence which is at least about 60% identical to the nucleotide sequence of SEQ ID NO:

1, 2, 3, or 4,

wherein the nucleic acid molecule is capable of directing mRNA expression from a promoterless reporter vector, or a complement thereof, or a cell comprising said nucleic acid molecule; and

b) determining whether mRNA expression or the polypeptide encoded by the cDNA is modulated,

thereby identifying a compound which is an EAAT2 expression promoting agent.

41. The method of claim 36, wherein the EAAT2 expression promoting agent is an antibiotic, an anti-hypertensive, a neurotransmitter, an antibacterial, an anti-inflammatory, a steroid derivative, or an anti-septic.

42. The method of claim 36, wherein the EAAT2 expression promoting agent comprises at least one structural element selected from heterocycles comprising at least one ring sulfur atom, tertiary amines, quaternary ammonium salts, steroids, polyols, polyketides, guanidine, urea, or arsenate.

43. The method of claim 42, wherein the EAAT2 expression promoting agent comprises at least one structural element selected from tertiary amines, quaternary ammonium salts, polyketides, steroidal ring systems and heterocycles having one or two rings, at least one sulfur ring atom and 0, 1, or 2 nitrogen ring atoms.

44. The method of claim 36 wherein the EAAT2 expression promoting agent increases EAAT2 production by 200% or more relative to non-regulated EAAT2 production.

45. The method of claim 44, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, vancomycin hydrochloride, thiaphenicol, ceftriaxone, ceftriaxone sodium, 1,3-dipropyl-8-cyclopentylxanthine, cephapirin sodium, actinospectacin, cefoperazole, cefoperazole sodium, acivicin, acetylcholine, chloramphenicol, vidarabine, atenolol, oxytetracycline, glafenine, oxymetazoline hydrochloride, gallamine, perillic acid (−

), amitriptyline hydrochloride, tetracaine hydrochloride, disopyramide phosphate, sisomicin sulfate, ketamine hydrochloride, xylazine, bicuculline, flurbiprofen, cefadroxil, bacampicillin hydrochloride, tiapride hydrochloride, norethindrone acetate, bergaptene, carisoprodol, citiolone, piroxicam, erythromycin ethylsuccinate, furegrelate sodium, albendazole, dihydrostreptomycin sulfate, aloin, fenoprofen, flutamide, ampicillin, ampicillin sodium, amprolium, sparteine sulfate, medroxyprogesterone acetate, alexidine hydrochloride, clindamycin hydrochloride, cephalothin, cephalothin sodium, daidzein, meclizine hydrochloride, lindane, bromopride, N-(3-trifluoromethylphenyl)piperazine hydrochloride, enoxolone, ipratropium bromide, bufexamac, gluconolactone, rifampin, hydroxychloroquine, coleoforsin, chloroxine, oxidopamine hydrochloride, camptothecin, nafcillin sodium, mianserin hydrochloride, acetarsol, prilocaine hydrochloride, deferoxamine mesylate, hexamethonium bromide, methenamine, paraxanthine, harmalol hydrochloride, pyrithione zinc, hydrocortisone butyrate, acetazolamide, aminoglutethimide, meclofenoxate hydrochloride, 2-phenpropylamino-5-nitrobenzoic acid, amiodarone hydrochloride, aconitine, hydroxyprogesterone caproate, and diosmin.

46. The method of claim 36, wherein the EAAT2 expression promoting agent increases EAAT2 production by 300% or more relative to non-regulated production.

47. The method of claim 46, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, vancomycin hydrochloride, thiaphenicol, ceftriaxone, 1,3-dipropyl-8-cyclopentylxanthine, cephapirin sodium, actinospectacin, cefoperazole, cefoperazole sodium, acivicin, acetylcholine, chloramphenicol, vidarabine, atenolol, oxytetracycline, glafenine, and oxymetazoline hydrochloride.

48. The method of claim 36, wherein the EAAT2 expression promoting agent increases EAAT2 production by 400% or more relative to non-regulated EAAT2 production.

49. The method of claim 48, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, quinapril, amoxicillin, pridinol methansulfonate, aklomide, and vancomycin hydrochloride.

50. The method of claim 36, wherein the EAAT2 expression promoting agent increases EAAT2 production by 600% or more relative to non-regulated EAAT2 production.

51. The method of claim 50, wherein the EAAT2 expression promoting agent is selected from the group consisting of penicillin V, penicillin V potassium, dequalinium chloride, and quinapril.

52. The method of claim 36, wherein the mammal is a primate.

53. The method of claim 52, wherein the mammal is a human.

54. The method of claim 36, wherein the extracellular glutamate concentration is reduced by at least about 50% relative non-regulated concentration.

56. The method of claim 36, wherein the EAAT2 expression promoting agent is a compound identified by a screening assay comprising the steps of:

a) contacting a cell that expresses EAAT2, with a test compound; and

b) determining whether expression of the EAAT2 in the cell is modulated in the presence of the test compound compared to in the absence of the test compound, thereby identifying a compound which modulates expression of the EAAT2.

57. The method of claim 36, wherein the EAAT2 expression promoting agent is a β

-lactam antibiotic.

58. The method of claim 36, wherein the EAAT2 expression promoting agent is a penicillin class, cephalosporin class, carbapenam class or monobactam class compound.

66. The method of claim 36, wherein the EAAT2 expression promoting agent is a compound identified by a screening assay comprising the steps of

a) contacting a cell that expresses EAAT2 with a test compound;

and

b) determining whether expression of the EAAT2 in the cell is modulated in the presence of the test compound compared to in the absence of the test compound, thereby identifying a compound which modulates expression of the EAAT2.

67. The method of claim 36, wherein the EAAT2 expression promoting agent is a β

-lactam antibiotic.

68. The method of claim 36, wherein the EAAT2 expression promoting agent is a penicillin class, cephalosporin class, carbapenam class or monobactam class compound.

59. A method of treating a mammal to modulate glutamate neurotransmission, the method comprising administering to the mammal a therapeutically effective amount of at least one EAAT2 expression promoting agent capable of increasing EAAT2 expression.

60. The method of claim 59, wherein the mammal has been identified as in need of such treatment.

61. The method of claim 59, wherein the mammal is in need of treatment for a condition that is associated with learning or memory.

62. The method of claim 61, wherein the administration is for enhancing learning, memory;

or cognition.