Compounds and methods for modulating the silencing of a polynucleotide of interest

A method for decreasing the level of a target polynucleotide in a cell, the method comprising administering to said cell a combination of an effective concentration of a polynucleotide comprising a heterologous silencing element and an effective amount of a compound of Formula (a)-(k), or a derivative or analog thereof:

• (a) triprolidine, derivatives, and analogs thereof:
• (b) dihydroptaeroxylin, derivatives, and analogs thereof:
• (c) fusidic acid, derivatives, and analogs thereof:
• (d) fenbufen, derivatives, and analogs thereof:
• (e) 3-beta-hydroxydeoxyodihydrodeoxygedunin, derivatives, and analogs thereof:
• (f) deferoxamine, derivatives, and analogs thereof:
• (g) thioguanine, derivatives, and analogs thereof:
• (h) 2-aminomethyl-1,4-benzodioxane, derivatives, and analogs thereof:
• (i) 3-alpha-hydroxy-3-deoxyangolensic acid methyl ester, derivatives, and analogs thereof:
• (j) lunarine, derivatives, and analogs thereof:
• and
• (k) bromocriptine, derivatives, and analogs thereof:
• wherein:
• each n is independently an integer from 1 to 20;
• a dashed line in a cyclic ring structure represents a bond that can be either present or absent in the ring;
• each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each R′1, R′2, and R′3 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each R″1, R″2, and R″3 is independently selected from the group consisting of —OR11 and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each X1, X2, X3, and X4 is independently selected from the group consisting of CH2, O, S, and NR′4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each X′1, X′2, and X′3 is independently N or CH;
• each X″ is independently halogen;
• and
• pharmaceutically and cosmetically acceptable salts thereof.

A method for decreasing the activity of a silencing element in a cell, the method comprising administering to the cell an effective amount of a compound of Formula (l)-(m) or a derivative or analog thereof:

• (l) colforsin, derivatives, and analogs thereof:
• and
• (m) metameconine, derivatives, and analogs thereof:
• wherein:
• n is an integer from 1 to 20;
• each R1, R2, R3, R4, R5, R6, R7, R8, and R9 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• R″1 and R″2 are each independently selected from the group consisting of —ORii and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• X1 is selected from the group consisting of CH2, O, S, and NR′4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• and
• pharmaceutically and cosmetically acceptable salts thereof.

A method for treating a disease state or unwanted condition, the method comprising administering to a subject in need of treatment thereof an effective amount of a polynucleotide comprising a silencing element, which, when administered to said subject, decreases the level of the target polynucleotide and an effective amount of a compound of Formula (a)-(k):

• (a) triprolidine, derivatives, and analogs thereof:
• (b) dihydroptaeroxylin, derivatives, and analogs thereof:
• (c) fusidic acid, derivatives, and analogs thereof:
• (d) fenbufen, derivatives, and analogs thereof:
• (e) 3-beta-hydroxydeoxyodihydrodeoxygedunin, derivatives, and analogs thereof:
• (f) deferoxamine, derivatives, and analogs thereof:
• (g) thioguanine, derivatives, and analogs thereof:
• (h) 2-aminomethyl-1,4-benzodioxane, derivatives, and analogs thereof:
• (i) 3-alpha-hydroxy-3-deoxyangolensic acid methyl ester, derivatives, and analogs thereof:
• (j) lunarine, derivatives, and analogs thereof:
• and
• (k) bromocriptine, derivatives, and analogs thereof:
• wherein:
• each n is independently an integer from 1 to 20;
• a dashed line in a cyclic ring structure represents a bond that can be either present or absent in the ring;
• each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each R′1, R′2, and R′3 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each R″1, R″2, and R″3 is independently selected from the group consisting of —OR11 and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each X1, X2, X3, and X4 is independently selected from the group consisting of CH2, O, S, and NR′4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each X′1, X′2, and X′3 is independently N or CH;
• each X″ is independently halogen;
• and
• pharmaceutically and cosmetically acceptable salts thereof.

A method for treating a disease state or unwanted condition, the method comprising administering to a subject in need of treatment thereof an effective amount of a polynucleotide comprising a silencing element, which, when administered to said subject, decreases the level of the target polynucleotide and an effective amount of at least one compound of Formula (l)-(m):

• (l) colforsin, derivatives, and analogs thereof:
• and
• (m) metameconine, derivatives, and analogs thereof:
• wherein:
• n is an integer from 1 to 20;
• each R1, R2, R3, R4, R5, R6, R7, R8, and R9 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• R″1 and R″2 are each independently selected from the group consisting of —OR11 and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• X1 is selected from the group consisting of CH2, O, S, and NR′4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• and
• pharmaceutically and cosmetically acceptable salts thereof.

A method for decreasing the level of a target polynucleotide in a cell, the method comprising administering to the cell a combination of an effective concentration of a polynucleotide comprising a heterologous silencing element and an effective amount of a first RNAi enhancer in combination with an effective amount of at least one second RNAi enhancer, wherein the combination of the first RNAi enhancer and at least one second RNAi enhancer synergistically decreases the level of the target polynucleotide in the cell, and wherein at least one of the first RNAi enhancer and the second RNAi enhancer is selected from the group consisting of a compound of Formula (a)-(k), and derivatives and analogs thereof, wherein said cell further comprises at least one heterologous silencing element:

• (a) triprolidine, derivatives, and analogs thereof:
• (b) dihydroptaeroxylin, derivatives, and analogs thereof:
• (c) fusidic acid, derivatives, and analogs thereof:
• (d) fenbufen, derivatives, and analogs thereof:
• (e) 3-beta-hydroxydeoxyodihydrodeoxygedunin, derivatives, and analogs thereof:
• (f) deferoxamine, derivatives, and analogs thereof:
• (g) thioguanine, derivatives, and analogs thereof:
• (h) 2-aminomethyl-1,4-benzodioxane, derivatives, and analogs thereof:
• (i) 3-alpha-hydroxy-3-deoxyangolensic acid methyl ester, derivatives, and analogs thereof:
• (j) lunarine, derivatives, and analogs thereof:
• and
• (k) bromocriptine, derivatives, and analogs thereof:
• wherein:
• each n is independently an integer from 1 to 20;
• a dashed line in a cyclic ring structure represents a bond that can be either present or absent in the ring;
• each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each R′1, R′2, and R′3 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each R″1, R″2, R″3, and R″4 is independently selected from the group consisting of —OR11 and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each X1, X2, X3, and X4 is independently selected from the group consisting of CH2, O, S, and NR′4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each X′1, X′2, and X′3 is independently N or CH;
• each X″ is independently halogen;
• and
• pharmaceutically and cosmetically acceptable salts thereof.

A method for decreasing the level of a target polynucleotide in a cell, the method comprising administering to said cell a combination of an effective concentration of a polynucleotide comprising a heterologous silencing element and an effective amount of a first RNAi enhancer in combination with an effective amount of at least one second RNAi enhancer, wherein the combination of the first RNAi enhancer and at least one second RNAi enhancer synergistically decreases the level of the target polynucleotide in the cell, and wherein at least one of the first RNAi enhancer and the second RNAi enhancer is selected from the group consisting of a compound of Formula (a)-(k), and derivatives and analogs thereof:

• (a) triprolidine, derivatives, and analogs thereof:
• (b) dihydroptaeroxylin, derivatives, and analogs thereof:
• (c) fusidic acid, derivatives, and analogs thereof:
• (d) fenbufen, derivatives, and analogs thereof:
• (e) 3-beta-hydroxydeoxyodihydrodeoxygedunin, derivatives, and analogs thereof:
• (f) deferoxamine, derivatives, and analogs thereof:
• (g) thioguanine, derivatives, and analogs thereof:
• (h) 2-aminomethyl-1,4-benzodioxane, derivatives, and analogs thereof:
• (i) 3-alpha-hydroxy-3-deoxyangolensic acid methyl ester, derivatives, and analogs thereof:
• (j) lunarine, derivatives, and analogs thereof:
• and
• (k) bromocriptine, derivatives, and analogs thereof:
• wherein:
• each n is independently an integer from 1 to 20;
• a dashed line in a cyclic ring structure represents a bond that can be either present or absent in the ring;
• each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each R′1, R′2, and R′3 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each R″1, R″2, and R″3 is independently selected from the group consisting of —OR11 and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each X1, X2, X3, and X4 is independently selected from the group consisting of CH2, O, S, and NR′4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each X′1, X′2, and X′3 is independently N or CH;
• each X″ is independently halogen;
• and
• pharmaceutically and cosmetically acceptable salts thereof.

A method for treating a disease state or unwanted condition, the method comprising administering to a subject in need of treatment thereof an effective amount of a polynucleotide comprising a silencing element, which, when administered to said subject, decreases the level of the target polynucleotide and an effective amount of a first RNAi enhancer in combination with an effective amount of at least one second RNAi enhancer, wherein the combination of the first RNAi enhancer and at least one second RNAi enhancer synergistically decreases the level of the target polynucleotide in the cell, and wherein at least one of the first RNAi enhancer and the second RNAi enhancer is selected from the group consisting of a compound of Formula (a)-(k), and derivatives and analogs thereof:

• (a) triprolidine, derivatives, and analogs thereof:
• (b) dihydroptaeroxylin, derivatives, and analogs thereof:
• (c) fusidic acid, derivatives, and analogs thereof:
• (d) fenbufen, derivatives, and analogs thereof:
• (e) 3-beta-hydroxydeoxyodihydrodeoxygedunin, derivatives, and analogs thereof:
• (f) deferoxamine, derivatives, and analogs thereof:
• (g) thioguanine, derivatives, and analogs thereof:
• (h) 2-aminomethyl-1,4-benzodioxane, derivatives, and analogs thereof:
• (i) 3-alpha-hydroxy-3-deoxyangolensic acid methyl ester, derivatives, and analogs thereof:
• (j) lunarine, derivatives, and analogs thereof:
• and
• (k) bromocriptine, derivatives, and analogs thereof:
• wherein:
• each n is independently an integer from 1 to 20;
• a dashed line in a cyclic ring structure represents a bond that can be either present or absent in the ring;
• each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each R′1, R′2, and R′3 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each R″1, R″2, and R″3 is independently selected from the group consisting of —OR11 and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• each X1, X2, X3, and X4 is independently selected from the group consisting of CH2, O, S, and NR′4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• each X′1, X′2, and X′3 is independently N or CH;
• each X″ is independently halogen;
• and
• pharmaceutically and cosmetically acceptable salts thereof.

A method for screening a compound of interest for the ability to modulate the activity of a heterologous silencing element comprising:

• a) providing a host cell that stably expresses a reporter gene, wherein said host cell further comprises at least one heterologous silencing element capable of inhibiting the expression of the reporter gene;
• b) administering to the cell a compound of interest in the presence of an RNAi enhancer;
• and
• c) measuring the expression of the reporter gene.

A method for modulating the activity of a silencing element in a cell, the method comprising administering to the cell an effective amount of at least one RNAi enhancer and an effective amount of at least one compound of Formula (l)-(m) or a derivative or analog thereof, to modulate the activity of said silencing element:

• (l) colforsin, derivatives, and analogs thereof:
• and
• (m) metameconine, derivatives, and analogs thereof:
• wherein:
• n is an integer from 1 to 20;
• each R1, R2, R3, R4, R5, R6, R7, R8, and R9 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• R″1 and R″2 are each independently selected from the group consisting of —OR11 and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• X1 is selected from the group consisting of CH2, O, S, and NR′4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• and
• pharmaceutically and cosmetically acceptable salts thereof.

A method for treating a disease state or unwanted condition, the method comprising administering to a subject in need of treatment thereof an effective amount of a polynucleotide comprising a silencing element, which, when administered to said subject, decreases the level of the target polynucleotide and an effective amount of at least one compound of Formula (l)-(m) and at least one RNAi enhancer:

• (l) colforsin, derivatives, and analogs thereof:
• and
• (m) metameconine, derivatives, and analogs thereof:
• wherein:
• n is an integer from 1 to 20;
• each R1, R2, R3, R4, R5, R6, R7, R8, and R9 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• R″1 and R″2 are each independently selected from the group consisting of —OR11 and —O(C═O)—R12, wherein R11 and R12 are selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, and substituted aryl;
• X1 is selected from the group consisting of CH2, O, S, and NR'4, wherein R′4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hydroxyl, and alkoxyl;
• and
• pharmaceutically and cosmetically acceptable salts thereof.