1. A recombinant polypeptide comprising an amino acid sequence at least 80% identical to SEQ ID NO: 1 wherein the amino acid sequence comprises one or more of the following features:
a) an amino acid with a negatively charged side chain at the position corresponding to Asn 13 of SEQ ID NO:1;
b) an amino acid with a negatively charged side chain at the position corresponding to Glu 17 of SEQ ID NO:1;
c) an amino acid with a negatively charged side chain at the position corresponding to Gln 64 of SEQ ID NO:1; or
d) an amino acid with a polar side chain at the position corresponding to Leu 67 of SEQ ID NO:1.
2. The polypeptide of claim 1, wherein the amino acid sequence comprises an amino acid with a negatively charged side chain at one or more of the positions corresponding to:
e) Glu 8 of SEQ ID NO:1;
f) Glu 12 of SEQ ID NO:1;
g) Glu 35 of SEQ ID NO:1; or
h) Asp 68 of SEQ ID NO:1.
3. The polypeptide of claim 1, wherein the polypeptide comprises at least two of said features.
4. The polypeptide of claim 1, wherein the polypeptide comprises an amino acid sequence at least 85% identical to SEQ ID NO: 1 or 3.
5. The polypeptide of claim 1, wherein the amino acid sequence comprises an Asp residue at the position corresponding to Asn 13 of SEQ ID NO: 1.
6. The polypeptide of claim 1, wherein the amino acid sequence comprises a Glu residue at the position corresponding to Glu 17 of SEQ ID NO: 1.
7. The polypeptide of claim 1, wherein the amino acid sequence comprises a Glu residue at the position corresponding to Gln 64 of SEQ ID NO: 1.
8. The polypeptide of claim 1, wherein the amino acid sequence comprises a Thr residue at the position corresponding to Leu 67 of SEQ ID NO: 1.
9. The polypeptide of claim 1, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 1.
10. The polypeptide of claim 1, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 3.
11. A method for binding uranyl comprising:
(a) contacting a solution comprising uranyl with a polypeptide comprising an amino acid sequence at least about 80% identical to SEQ ID NO: 1 or 3, thereby binding the uranyl to the polypeptide.
12. The method of claim 11, wherein the solution comprises uranyl having a 238U, 235U or 233U uranium isotope.
13. The method of claim 11, wherein the solution comprises carbonate.
14. The method of claim 11, wherein the polypeptide binds to uranyl with a KD of between about 1.8 nM and about 1.0 fM.
15. The method of claim 11, further comprising isolating or concentrating the polypeptide bound to uranyl.
16. The method of claim 11, further comprising eluting uranyl from the polypeptide.
17. The method of claim 16, wherein eluting comprises heating the polypeptide.
18. A method for reducing the level of uranium in a solution comprising:
(a) contacting a solution comprising uranyl with the polypeptide of claim 1 thereby binding the uranyl to the polypeptide; and
(b) purifying the polypeptide away from the solution thereby reducing the level of uranium in the solution.
19. A method for binding uranyl comprising:
(a) contacting a solution comprising uranyl with the polypeptide of claim 1, thereby binding the uranyl to the polypeptide.
20. The method of claim 19, wherein the polypeptide binds to uranyl with a KD of between about 100 nM and about 0.1 fM.
The claims below are in addition to those above.
All refrences to claims which appear below refer to the numbering after this setence.
1. A compound capable of modifying ion channels, in vivo, having a formula:
or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein:
wherein A is N, CR4, a carbon atom bound to L, or is not an atom;
one of W, Z, B, Y and X is a carbon atom bound to L if A is not an atom, another of W, Z, B, Y and X is a carbon atom bound to G, and each of the remaining W, Z, B, Y and X is independently N or CR4;
L is substituted or unsubstituted -(C-C)-, -(CR5=CR6)- or -(C\u2261C)-;
G is C=O, C=S or SO2;
R1 is substituted or unsubstituted aliphatic, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl;
R2 is hydrogen or substituted or unsubstituted alkyl;
R3 is substituted or unsubstituted aliphatic, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl;
each R4 is independently hydrogen, alkyl, substituted or unsubstituted alkyl, acyl, acylamino, alkylamino, alkylthio, alkoxy, alkoxycarbonyl, alkylarylamino, arylalkyloxy, amino, aryl, arylalkyl, sulfoxide, sulfone, sulfanyl, aminosulfonyl, arylsulfonyl, sulfuric acid, sulfuric acid ester, dihydroxyphosphoryl, aminohydroxyphosphoryl, azido, carboxy, carbamoyl, carboxyl, cyano, cycloheteroalkyl, dialkylamino, halo, heteroaryloxy, heteroaryl, heteroalkyl, hydroxyl, nitro or thio; and
each of R5 and R6 is independently H, halo, or substituted or unsubstituted aliphatic, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl.
2. A compound according to claim 1 wherein the compound is of the formula:
and all substituents may be as defined in the above.
3. A compound according to claim 2 wherein
L is substituted or unsubstituted -(C-C)-, -(CR5=CR6)- or -(C\u2261C)-;
G is C=O;
R1 is substituted or unsubstituted aliphatic, alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl;
R2 is hydrogen;
R3 is substituted or unsubstituted aliphatic or alkyl.
4. A compound according to claim 3 wherein R3 is CF3, n-propyl, or a group of formula:
wherein R2\u2019 is hydrogen or alkyl; and wherein two R2\u2019 may join together to form a cycloalkyl or cycloheteroalkyl ring of 3-8 atoms; provided at least two of R2\u2019 are alkyl.
5. A compound according to claim 4 wherein the compound is of the formula:
and all substituents may be as defined in the above.
6. A compound according to claim 5 wherein R1 is substituted alkyl.
7. A compound according to claim 6 wherein R1 is \u2013(CR2
2)x- R4\u2019; R2 is hydrogen or alkyl; R4\u2019 is R4; R4 is as described in claim 1; and x is an integer from 1-3.
8. A compound according to claim 7 wherein R4\u2019 is selected from t-butyl, aryl, cycloalkyl, cycloheteroalkyl and heteroaryl.
9. A compound according to claim 8 wherein R4\u2019 is substituted or unsubstituted phenyl, or naphthalenyl.
10. A compound according to claim 8 wherein R4\u2019 is substituted or unsubstituted cyclopropyl, cyclopentyl or cyclohexyl.
11. A compound according to claim 8 wherein R4\u2019 is substituted or unsubstituted pyrrolidinyl, piperidinyl, or morpholinyl.
12. A compound according to claim 8 wherein R4\u2019 is substituted or unsubstituted pyridinyl or pyrimidinyl.
13. A compound according to claim 8 wherein R4\u2019 is substituted or unsubstituted furanyl, imidazolyl, thiophenyl, pyrazolyl, or thiazolyl.
14. A compound according to claim 8 wherein R4\u2019 is substituted or unsubstituted benzodioxanyl, benzopyranyl, indolyl, indazolyl, methylenedioxyphenyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, dihydroquinolinyl, or dihydroisoquinolinyl.
15. A compound according to claim 8 wherein R4\u2019 is t-Bu.
16. A compound according to any of claims 8-15 wherein x is 1 or 2.
17. A compound according to claim 6 wherein R1 is t-Bu.
18. A compound according to claim 5 wherein R1 is substituted or unsubstituted cycloalkyl, cycloheteroalkyl or heteroaryl.
19. A compound according to claim 18 wherein R1 is substituted or unsubstituted cyclopropyl or cyclopentyl.
20. A compound according to claim 18 wherein R1 is substituted or unsubstituted pyrrolidinyl, piperidinyl, or morpholinyl.
21. A compound according to claim 18 wherein R1 is substituted or unsubstituted pyridinyl or pyrimidinyl.
22. A compound according to claim 18 wherein R1 is substituted or unsubstituted furanyl, imidazolyl, thiophenyl, pyrazolyl, or thiazolyl.
23. A compound according to claim 18 wherein R1 is substituted or unsubstituted benzodioxanyl, benzopyranyl, indolyl, indazolyl, methylenedioxyphenyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, dihydroquinolinyl, or dihydroisoquinolinyl.
24. A compound according to claim 5 wherein R1 is substituted or unsubstituted aryl.
25. A compound according to claim 24 wherein R1 is substituted or unsubstituted phenyl, naphthalenyl, 2-biphenyl or 4-biphenyl.
26. A compound according to claim 25 wherein the compound is of the formula:
wherein R1\u2019 is R4; and x is selected from 1-5.
27. A compound according to claim 26 wherein x is 1.
28. A compound according to claim 27 wherein R1\u2019 is selected from the group consisting of methyl, isopropyl, t-butyl, cyano, trifluoroalkyl, halo, nitro, methoxy, trifluoromethoxy, amino, alkylamino, dialkylamino, phenyl, SO2Me, SO2CF3, SO2NMe2, and carboxy.
29. A compound according to claim 28 wherein the R1\u2019 substitution is at the 4-position.
30. A compound according to any one of claims 5-29 wherein each of W, X, Y and Z is CR4.
31. A compound according to any one of claims 5-29 wherein W is N and each of X, Y and Z is CR4.
32. A compound according to any one of claims 5-29 wherein Y is N and each of W, X and Z is CR4.
33. A compound according to any one of claims 5-29 wherein two of W, X, Y and Z are Ns.
34. A compound according to any one of claims 30-33 wherein L is
(CR5=CR6).
35. A compound according to claim 34 wherein R5 and R6 are both hydrogen.
36. A compound according to claim 34 wherein R5 is methyl and R6 is hydrogen.
37. A compound according to claim 34 wherein R5 is hydrogen and R6 is methyl.
38. A compound according to any one of claims 30-33 wherein L is
or (C\u2261C).
39. A compound according to any one of claims 34-38 wherein R3 is t-Bu or i-Pr.
40. A compound according to any one of claims 34-38 wherein R3 is CF3.
41. A compound according to claim 18 wherein the compound is of Table 1 and wherein the compound ID No. is 5, 13, 17, 35, 81, 87, 111, 117, 142, 143, 185, 196, 208, 211, 213, 238, 239, 275, or 305, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
42. A compound according to claim 18 wherein the compound is of Table 1 and wherein the compound ID No. is 11, 20, 24, 98, 116, 121, 194, 217, 286, or 304, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
43. A compound according to claim 24 wherein the compound is of Table 1 and wherein the compound ID No. is 2, 3, 15, 30, 31, 32, 33, 37, 39, 40, 41, 42, 43, 44, 45, 47, 61, 62, 66, 78, 130, 132, 136, 145, 148, 150, 151, 153, 156, 158, 169, 171, 172, 225, 226, 227, 228, 230, 233, 242, 244, 245, 246, 247, 248, 249, 252, 313, 330, 331, 332, 334, 336, 337, 338, or 358, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
44. A compound according to claim 24 wherein the compound is of Table 1 and wherein the compound ID No. is 48, 52, 53, 64, 67, 68, 70, 71, 72, 74, 75, 76, 77, 105, 106, 166, 168, 202, 203, 234, 256, 259, 260, 261, 324, 341, 344, 345, or 351, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
45. A compound according to claim 24 wherein the compound is of Table 1 and wherein the compound ID No. is 54, 55, 204, or 205, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
46. A compound according to claim 24 wherein the compound is of Table 1 and wherein the compound ID No. is 8, 12, 56, 65, 181, 207, 214, 237, or 287, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
47. A compound according to claim 5 wherein the compound is of Table 1 and wherein the compound ID No. is 6, 9, 14, 16, 26, 29, 49, 60, 80, 88, 93, 95, 109, 112, 120, 123, 125, 126, 127, 139, 164, 179, 183, 190, 191, 192, 193, 197, 198, 206, 209, 216, 218, 219, 223, 224, 236, 267, 281, 283, 308, 315, 316, 356, or 357, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
48. A compound according to claim 38 wherein the compound is of Table 1 and wherein the compound ID No. is 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, or 412, or a pharmaceutically acceptable salt or prodrug thereof.
49. A compound according to claim 38 wherein the compound is of Table 1 and wherein the compound ID No. is 411, 413-439, or 440, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
50. A compound according to claim 40 wherein the compound is of Table 1 and wherein the compound ID No. is 388, 389, 390, 391, 392, 470, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, or 405, or a pharmaceutically acceptable salt, solvate or prodrug thereof; and stereoisomers thereof.
51. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of a compound of any of claims 1-50.
52. The pharmaceutical composition of claim 51 wherein the carrier is a parenteral carrier.
53. The pharmaceutical composition of claim 51 wherein the carrier is an oral carrier.
54. The pharmaceutical composition of claim 51 wherein the carrier is a topical carrier.
55. A method for preventing, treating, ameliorating or managing a disease or condition which comprises administering to a patient in need of such prevention, treatment, amelioration or management, a prophylactically or therapeutically effective amount of a compound of any of claims 1-50, or the pharmaceutical composition of any of claims 51-54.
56. The method of claim 55 wherein the disease or condition is a pain condition.
57. The method of claim 55 wherein the disease or condition is an autoimmune disease.
58. The method of claim 55 wherein the disease or condition is an inflammatory disease or condition.
59. The method of claim 55 wherein the disease or condition is a neurological or neurodegenerative disease or condition.
60. A method for preventing, treating, ameliorating or managing a disease or condition, which comprises administering to a patient in need of such prevention, treatment, amelioration or management a prophylactically or therapeutically acceptable amount of a compound of any of claims 1-50, or the pharmaceutical composition of any of claims 51-54, wherein the disease or condition is: pain including acute, inflammatory and neuropathic pain; chronic pain; dental pain; headache including migraine, cluster headache and tension headache; Parkinson\u2019s disease; Alzheimer\u2019s disease; multiple sclerosis; diseases and disorders mediated by or result in neuroinflammation, traumatic brain injury, stroke, or encephalitis; centrally-mediated neuropsychiatric diseases and disorders including depression, mania, bipolar disease, anxiety, schizophrenia, eating disorders, sleep disorders and cognition disorders; epilepsy and seizure disorders; prostate, bladder and bowel dysfunction, urinary incontinence, urinary hesitancy, rectal hypersensitivity, fecal incontinence, benign prostatic hypertrophy and inflammatory bowel disease; respiratory and airway disease and disorders including allergic rhinitis, asthma and reactive airway disease and chronic obstructive pulmonary disease; diseases and disorders mediated by or result in inflammation including arthritis, rheumatoid arthritis and osteoarthritis; myocardial infarction; autoimmune diseases and disorders; uveitis and atherosclerosis; itchpruritus, psoriasis; alopecia (hair loss); obesity; lipid disorders; cancer; high blood pressure; spinal cord injury; or renal disorders.
61. The method of claim 60 wherein the disease or condition is Parkinson\u2019s disease.
62. The method of claim 60 wherein the disease or condition is Alzheimer\u2019s disease.
63. The method of claim 60 wherein the disease or condition is traumatic brain injury.
64. The method of claim 60 wherein the disease or condition is stroke.
65. The method of claim 60 wherein the disease or condition is pain.
66. The method of claim 60 wherein the disease or condition is neuropathic pain.
67. A method for preparing a compound of any of claims 1-66 which comprises contacting a compound of the formula R3-L-Cy-COCl or R3-L-Cy-SO2Cl with a compound of the formula R1R2NH under conditions sufficient to form a compound according to any of claims 1-50; and wherein Cy is aryl or heteroaryl.
68. A method of treating a mammal suffering from at least one symptom selected from the group consisting of symptoms of exposure to capsaicin, symptoms of burns or irritation due to exposure to heat, symptoms of burns or irritation due to exposure to light, symptoms of burns, bronchoconstriction or irritation due to exposure to tear gas, and symptoms of burns or exposure irritation due to exposure to acid which comprises administering to the mammal an effective disease-treating or condition-treating amount of a compound of any of claims 1-50, or the pharmaceutical composition of any of claims 51-54.
69. The method of claim 68 wherein the pain is associated with a condition selected from the group consisting of postmastectomy pain syndrome, stump pain, phantom limb pain, oral neuropathic pain, Charcot\u2019s pain, toothache, venomous snake bite, spider bite, insect sting, postherpetic neuralgia, diabetic neuropathy, reflex sympathetic dystrophy, trigeminal neuralgia, osteoarthritis, rheumatoid arthritis, fibromyalgis, Guillain-Barre syndrome, meralgia paresthetica, burning-mouth syndrome, bilateral peripheral neuropathy, causalgia, sciatic neuritis, peripheral neuritis, polyneuritis, segmental neuritis, Gombault\u2019s neuritis, neuronitis, cervicobrachial neuralgia, cranial neuralgia, egniculate neuralgia, glossopharyngial neuralgia, migranous neuralgia, idiopathic neuralgia, intercostals neuralgia, mammary neuralgia, mandibular joint neuralgia, Morton\u2019s neuralgia, nasociliary neuralgia, occipital neuralgia, red neuralgia, Sluder\u2019s neuralgia, splenopalatine neuralgia, supraorbital neuralgia, vidian neuralgia, sinus headache, tension headache, labor, childbirth, intestinal gas, menstruation, cancer, and trauma.
70. A compound of any of claims 1-50 for use as a pharmaceutical.
71. A compound of any of claims 1-50 for use as a pharmaceutical for the treatment of the conditions as set forth in any of claims 55-66, 68, or 69.
72. Use of a compound of claims 1-50 in the preparation of a pharmaceutical composition for the treatment of the conditions as set forth in any of claims 55-66, 68, or 69.