Patent Grant
8148577
This invention pertains to polyamine compounds, including polyamine/guanidine and polyamine/biguanide compounds, useful for treatment of cancer and/or parasitic infections, and for inhibition of lysine-specific demethylase.
Polyamines are found in both eukaryotic and prokaryotic cells and figure prominently in regulation of the cell cycle and cell division. Agents specifically targeting polyamine biosynthesis, such as polyamine analogs, have been shown to have therapeutic effect in treatment of cancer, parasitic diseases, and other indications. These antiproliferatvie effects have been demonstrated to be, in part, a result of agent-induced decreases in the natural intracellular polyamines resulting from inhibition, down-regulation of polyamine biosynthesis and/or up regulation of polyamine catabolism. See, e.g., Wang and Casero, J, Biochem. 139:17 (2006); Casero et al., Proc. West. Pharmacol. Soc. 48:24 (2005); Casero et al., J. Med. Chem. 44:1 (2001); U.S. Pat. Nos. 5,889,061, 6,392,098, and 6,794,545; U.S. Patent Application Publication Nos. 2003/0072715, 2003/0195377, and International Patent Applications WO 98/17624, WO 00/66587, WO 02/10142, and WO 03/050072. Bi et al., Bioorgan. Med. Chem. Letters 16:3229 (2006) discuss novel alkylpolyaminoguanidines and alkylpolyaminobiguanides with potent antitrypanosomal activity.
The enzyme lysine-specific demethylase-1 (LSD1) has been shown to play an important role in regulation of gene expression; see Shi et al., Cell 119:941 (2004). WO 2006/071608 discusses certain methods involving lysine-specific demethylase-1. In view of the importance of gene regulation in areas such as cancer therapy and cancer prophylaxis, inhibitors of LSD1 are of great interest in the treatment and prevention of cancer and uncontrolled cell growth.
The invention embraces polyamine, polyamine/guanidine, and polyamine/biguanide compounds, and uses of those compounds for treatment and prevention of cancer. The invention also embraces uses of those compounds for inhibition of lysine-specific demethylase-1, and treatment of diseases involving lysine-specific demethylase-1.
In one embodiment, the invention embraces compounds of the formula (M):
E-X-A-NH—B—NH-A-X-E (M)
where each E is independently selected from hydrogen, C1-C8 substituted or unsubstituted alkyl, C4-C15 substituted or unsubstituted cycloalkyl, C3-C15 substituted or unsubstituted branched alkyl, C6-C20 substituted or unsubstituted aryl or heteroaryl, C7-C24 substituted or unsubstituted aralkyl or heteroalkyl or heteroaralkyl, C3-C24 substituted or unsubstituted heteroaryl; each A is independently a C1-C8 n-alkyl; B is independently selected from C1-C12 n-alkyl or C3-C8 cycloalkyl; and each X is independently selected from —NH—, —NH—C(═NH)—NH—, and —NH—C(═NH)—NH—C(═NH)—NH—; and all salts, solvates, hydrates, and stereoisomers thereof.
In another embodiment, B is independently selected from C1-C8 n-alkyl.
In another embodiment, at least one X is selected from —NH—C(═NH)—NH— and —NH—C(═NH)—NH—C(═NH)—NH—. In another embodiment, at least one X is —NH—C(═NH)—NH—. In another embodiment, at least one X is —NH—C(═NH)—NH—C(═NH)—NH—. In another embodiment, each X is independently selected from —NH—C(═NH)—NH— and —NH—C(═NH)—NH—C(═NH)—NH—. In another embodiment, both X groups are —NH—C(═NH)—NH—. In another embodiment, both X groups are —NH—C(═NH)—NH—C(═NH)—NH—. In another embodiment, one X is —NH—C(═NH)—NH— and another X is —NH—C(═NH)—NH—C(═NH)—NH—.
In one embodiment, the invention embraces polyamine/guanidine or N-alkylated polyamine/guanidine compounds, such as a polyaminobisguanidine or polyaminobiguanide or N-alkylated variation thereof. An N-alkylated polyaminoguanidine intends a polyaminoguanidine wherein the imine nitrogen of the guanidine is alkylated, such as in a 2-methylguanadine derivative. In one embodiment, each A is —(CH2)3— and B is —(CH2)4—. In another embodiment, each A is —(CH2)3— and B is —(CH2)7—.
In one embodiment, the compound is a polyaminoguanidine of the formula (I):
or a salt, solvate, or hydrate thereof, wherein n is an integer from 1 to 12, m and p are independently an integer from 1 to 5, q is 0 or 1, each R1 is independently selected from the group consisting of C1-C8 substituted or unsubstituted alkyl, C4-C15 substituted or unsubstituted cycloalkyl, C3-C15 substituted or unsubstituted branched alkyl, C6-C20 substituted or unsubstituted aryl, C6-C20 substituted or unsubstituted heteroaryl, C7-C24 substituted or unsubstituted aralkyl, and C7-C24 substituted or unsubstituted heteroaralkyl, and each R2 is independently selected from hydrogen or a C1-C8 substituted or unsubstituted alkyl.
In one embodiment, the compound is of the formula (I) wherein at least one or both R1 is a C6-C20 substituted or unsubstituted aryl, such as a single ring substituted or unsubstituted aryl, including without limitation, substituted or unsubstituted phenyl. In one embodiment, the compound is of the formula (I) and each R1 is phenyl. In one embodiment, q is 1, m and p are 3, and n is 4. In another embodiment, q is 1, m and p are 3, and n is 7.
In one embodiment, the compound is of the formula (I) wherein at least one or both R1 is a C8-C12 or a C1-C8 substituted or unsubstituted alkyl, such as a linear alkyl. One or both R1 may be a C1-C8 substituted or unsubstituted linear alkyl, such as methyl or ethyl. In one embodiment, each R1 is methyl. Each or both R1 may comprise or be a C4-C15 cycloalkyl group, such as a cycloalkyl group containing a linear alkyl group, where the cycloalkyl group is connected to the molecule either via its alkyl or cycloalkyl moiety. For instance, each or both R1 may be cyclopropylmethyl or cyclohexylmethyl. In one embodiment, one R1 is cyclopropylmethyl or cyclohexylmethyl and the other R1 is a linear alkyl group, such as a linear C1-C8 unsubstituted alkyl group, including without limitation an ethyl group. In one embodiment, R1 is a C3-C15 branched alkyl group such as isopropyl. When R1 is a C1-C8 substituted alkyl, the substituted alkyl may be substituted with any substituent, including a primary, secondary, tertiary or quaternary amine. Accordingly, in one embodiment, R1 is a C1-C8 alkyl group substituted with an amine such that R1 may be e.g., alkyl-NH2 or an alkyl-amine-alkyl moiety such as —(CH2)yNH(CH2)zCH3 where y and z are independently an integer from 1 to 8. In one embodiment, R1 is —(CH2)3NH2.
In one embodiment, the compound is of the formula (I) where at least one R1 is a C7-C24 substituted or unsubstituted aralkyl, which in one embodiment is an aralkyl connected to the molecule via its alkyl moiety (e.g., benzyl). In one embodiment, each R1 is an aralkyl moiety wherein the alkyl portion of the moiety is substituted with two aryl groups and the moiety is connected to the molecule via its alkyl group. For instance, in one embodiment at least one or both R1 is a C7-C24 aralkyl wherein the alkyl portion is substituted with two phenyl groups, such as when R1 is 2,2-diphenylethyl or 2,2-dibenzylethyl. In one embodiment, each R1 of formula (I) is 2,2-diphenylethyl and n is 1, 2 or 5. In one embodiment, each R1 of formula (I) is 2,2-diphenylethyl, n is 1, 2 or 5 and m and p are each 1.
In one embodiment, at least one R1 is hydrogen. When at least one R1 is hydrogen, the other R1 may be any moiety listed above for R1, including an aryl group such as benzyl.
Any of the compounds of formula (I) listed above include compounds where at least one or both of R2 is hydrogen or a C1-C8 substituted or unsubstituted alkyl. In one embodiment, each R2 is an unsubstituted alkyl such as methyl. In another embodiment, each R2 is hydrogen.
Any of the compounds of formula (I) listed above may be compounds where q is 1 and m and p are the same. Accordingly, the polyaminoguanidines of formula (I) may be symmetric with reference to the polyaminoguanidine core (e.g., excluding R1). Alternatively, the compounds of formula (I) may be asymmetric, e.g., when q is 0. In one embodiment, m and p are 1. In one embodiment, q is 0. In one embodiment, n is an integer from 1 to 5.
It is understood and clearly conveyed by this disclosure that each R1, R2, m, n, p and q disclosed in reference to formula (I) intends and includes all combinations thereof the same as if each and every combination of R1, R2, m, n, p and q were specifically and individually listed.
Representative compounds of the formula (I) include, e.g.:
In one embodiment, the compound is a polyaminobiguanide or N-alkylated polyaminobiguanide. An N-alkylated polyaminobiguanide intends a polyaminobiguanide wherein at least one imine nitrogen of at least one biguanide is alkylated. In one embodiment, the compound is a polyaminobiguanide of the formula (II):
or a salt, solvate, or hydrate thereof, wherein n is an integer from 1 to 12, m and p are independently an integer from 1 to 5, q is 0 or 1, each R1 is independently selected from the group consisting of C1-C8 substituted or unsubstituted alkyl, C6-C20 substituted or unsubstituted aryl, C6-C20 substitute or unsubstituted heteroaryl, C7-C24 substituted or unsubstituted aralkyl, and C7-C24 substituted or unsubstituted heteroaralkyl and each R2 is independently hydrogen or a C1-C8 substituted or unsubstituted alkyl.
In one embodiment, at least one or each R1 is a C1-C8 substituted or unsubstituted alkyl, such as those listed above in reference to formula (I). For instance, when R1 is a C1-C8 substituted alkyl, the substituted alkyl may be substituted with any substituent, including a primary, secondary, tertiary or quaternary amine. Accordingly, in one embodiment, R1 is a C1-C8 alkyl group substituted with an amine such that R1 may be e.g., alkyl-NH2 or an alkyl-amine-alkyl moiety such as —(CH2)yNH(CH2)zCH3 where y and z are independently an integer from 1 to 8. In one embodiment, R1 is —(CH2)3NH2. R1 may also be a C4-C15 substituted or unsubstituted cycloalkyl or a C3-C15 substituted or unsubstituted branched alkyl, such as described for formula (I) above. In one embodiment, at least one or each R1 is a C6-C20 substituted or unsubstituted aryl, such as those listed above in reference to formula (I), In one embodiment, q is 1, m and p are 3, and n is 4. In another embodiment, q is 1, m and p are 3, and n is 7.
In one embodiment, the compound is of the formula (II) where at least one or both R1 is a C7-C24 substituted or unsubstituted aralkyl, which in one embodiment is an aralkyl connected to the molecule via its alkyl moiety. In one embodiment, each R1 is an aralkyl moiety wherein the alkyl portion of the moiety is substituted with one or two aryl groups and the moiety is connected to the molecule via its alkyl moiety. For instance, in one embodiment at least one or both R1 is an aralkyl wherein the alkyl portion is substituted with two phenyl or benzyl groups, such as when R1 is 2,2-diphenylethyl or 2,2-dibenzylethyl. In one embodiment, each R1 of formula (II) is 2,2-diphenylethyl and n is 1, 2 or 5. In one embodiment, each R1 of formula (II) is 2,2-diphenylethyl and n is 1, 2 or 5 and m and p are each 1.
Any of the compounds of formula (II) listed above include compounds where at least one or both of R2 is hydrogen or a C1-C8 substituted or unsubstituted alkyl. In one embodiment, each R2 is an unsubstituted alkyl, such as methyl. In another embodiment, each R2 is a hydrogen.
Any of the compounds of formula (II) listed above include compounds where q is 1 and m and p are the same. Accordingly, the polyaminobiguanides of formula (II) may be symmetric with reference to the polyaminobiguanide core (e.g., excluding R1). Alternatively, the compounds of formula (II) may be asymmetric, e.g., when q is 0. In one embodiment, m and p are 1. In one embodiment, q is 0. In one embodiment, n is an integer from 1 to 5. In one embodiment, q, m and p are each 1 and n is 1, 2 or 5.
It is understood and clearly conveyed by this disclosure that each R1, R2, m, n, p and q disclosed in reference to formula (II) intends and includes all combinations thereof the same as if each and every combination of R1, R2, m, n, p and q were specifically and individually listed.
Representative compounds of the formula (II) include, e.g.:
In one embodiment, the compound is a polyamine. In one embodiment, the polyamine is of the formula (III):
or a salt, solvate, or hydrate thereof, wherein n is an integer from 1 to 12; m and p are independently an integer from 1 to 5; R3 and R4 are independently selected from the group consisting of hydrogen, C1-C8 substituted or unsubstituted alkyl, C5-C20 substituted or unsubstituted aryl and C7-C24 substituted or unsubstituted aralkyl; R5, R9, R6, R7 and R8 are independently selected from the group consisting of hydrogen and C1-C8 substituted or unsubstituted alkyl; and wherein either m and p are not the same integer or at least one of R5, R9, R6, R7 and R8 is a C1-C8 substituted or unsubstituted alkyl.
In one embodiment, R9 is a C1-C8 substituted or unsubstituted alkyl. When R9 is a C1-C8 substituted alkyl, the substituted alkyl may be substituted with any substituent, including a primary, secondary, tertiary or quaternary amine. Accordingly, in one embodiment, R9 is a C1-C8 alkyl group substituted with an amine such that R9 may be e.g., alkyl-NH2 or an alkyl-amine-alkyl moiety such as —(CH2)yNH(CH2)zCH3 where y and z are independently an integer from 1 to 8. In one embodiment, R9 is —(CH2)3NHCH2CH3.
In one embodiment, one or both of R3 and R4 is hydrogen. If only one of R3 and R4 is hydrogen, the R3 or R4 that is not hydrogen may be any moiety described herein, such as a C1-C8 substituted or unsubstituted alkyl group, including a cyclic alkyl group such as cyclopropylmethyl or cycloheptylmethyl.
In one embodiment, one or both of R3 and R4 is a C1-C8 substituted or unsubstituted alkyl, including without limitation a substituted or unsubstituted n-alkyl (such as n-pentyl), substituted or unsubstituted branched (C3-C8) alkyl (such as 2-methylbutyl) or substituted or unsubstituted (C3-C8) cycloalkyl (such as cyclohexylmethyl). Larger chain alkyl (linear, branched and cyclic) are also considered, such as a C9-C15 substituted or unsubstituted alkyl. Where one or both of R3 and R4 is a C1-C8 substituted or unsubstituted n-alkyl, the moiety may be any n-alkyl, such as methyl or ethyl. In one embodiment, both R3 and R4 are a C1-C8 substituted or unsubstituted alkyl, wherein one of R3 and R4 is an n-alkyl moiety and the other is a cyclic moiety, which is understood to contain at least three carbon atoms. Alternatively, both R3 and R4 may be a C1-C8 substituted or unsubstituted n-alkyl. When one or both of R3 and R4 is a substituted alkyl, whether linear, branched or cyclic, the alkyl may be substituted with one or more substituents such as those listed under “Substituted alkyl” and includes alkyl substituted with any halogen, such as a monohaloalkyl, dihaloalkyl, trihaloalkyl or multihaloalkyl, including a perhalooalkyl, for example, perfluoroalkyl and percholoralkyl, such as trifluoromethyl or pentachloroethyl.
In one embodiment, one or both of R3 and R4 is a C6-C20 substituted or unsubstituted aryl. In one embodiment, one or both of R3 and R4 is a C6-C20 substituted aryl, which aryl groups may be substituted with one or more substituents such as those listed under “Substituted aryl.” In one embodiment, one or both of R3 and R4 is a C6-C20 substituted aryl, which aryl groups may be substituted with one or more alkyoxy (such as —OCH3), alkyl (including a branched alkyl such as tert-butyl), or halo groups (such as fluoro). In one embodiment, one or both of R3 and R4 is a halo-substituted aryl or a halo-substituted aralkyl, such as 2,4,5-trifluorophenyl or 2,4,5-trifluorobenzyl. In one embodiment, one or both of R3 and R4 is a di-alkyl-monoalkoxy-substituted aryl or aralkyl, such as 4,5-di-tert-butyl-2-methoxybenzyl or 4,5-di-tert-butyl-2-methoxyphenyl.
In one embodiment, one or both of R3 and R4 is a C7-C24 substituted or unsubstituted aralkyl or heteroaralkyl such as an aralkyl or heteroaralkyl connected to the molecule via its alkyl moiety. In one embodiment, one or both of R3 and R4 is a substituted aralkyl or heteroaralkyl connected to the molecule via its alkyl moiety. A substituted aralkyl may be substituted with one or more substituents such as those listed under “Substituted aralkyl” and a substituted heteroaralkyl may be substituted with one or more substituents such as those listed under “Substituted heteroaralkyl.” In one embodiment, one or both of R3 and R4 is a substituted heteroaralkyl having at least one nitrogen atom. In one embodiment, one or both of R3 and R4 is a single ring heteroaralkyl having at least one nitrogen atom. In one embodiment, one or both of R3 and R4 is 1-(2-N-methylpyrrolyl)-methyl.
In one embodiment, at least 1 or at least 2 or at least 3 of R5, R9, R6, R7 and R8 is a C1-C8 substituted or unsubstituted alkyl. R5, R9, R6, R7 and R8 may be a C1-C8 substituted or unsubstituted alkyl. In one embodiment at least 1 or at least 2 or at least 3 of R5, R9, R6, R7 is a C1-C8 unsubstituted n-alkyl, such as methyl or ethyl. In one embodiment, both R6 and R5 are methyl or ethyl. In one embodiment, at least one R7 and R8 is methyl or ethyl. In one embodiment, R7 is methyl.
It is understood and clearly conveyed by this disclosure that each R3, R4, R5, R9, R6, R7, R8, m, n, y, z and p disclosed in reference to formula (III) intends and includes all combinations thereof the same as if each and every combination of R3, R4, R5, R9, R6, R7, R8, m, n, y, z and p were specifically and individually listed.
Representative compounds of the formula (III) include, e.g.:
In one embodiment, the polyamine is of the formula (IV):
or a salt, solvate, or hydrate thereof, wherein A, R10 and R11 are independently (CH2)n or ethene-1,1-diyl; n is an integer from 1 to 5; R12 and R13 are independently selected from the group consisting of hydrogen, C2-C8 substituted or unsubstituted alkenyl and C1-C8 substituted or unsubstituted alkyl; and at least one of A, R10, R11, R12 and R13 comprises an alkenyl moiety. In another embodiment, when any one or more of A, R10, and R11 is alkenyl, the alkene portion branches off the direct chain connecting the nitrogen atoms; that is, no more than one sp2-hybridized carbon occurs in the carbon nodes along the shortest path from one nitrogen flanking A, R10, and/or R11 to the other flanking nitrogen. For example, when A is ethene, the segment containing A is of the form —CH2C(═CH2)—CH2— and the three nodes in the shortest carbon path between the nitrogens containing the A moiety has only one sp2-hybridized carbon. When A is propene, the segment containing A can be of the form —CH2C(═CHCH3)—CH2— or —CH2C(—CH═CH2)—CH2—.
In one embodiment, A is (CH2)n and n is 1. In one embodiment, A is ethene-1,1-diyl. In one embodiment, A is (CH2)n and one or both of R12 and R13 comprises an alkenyl moiety, such as propen-2-yl.
In one embodiment at least one or both of R10 and R11 is ethene-1,1-diyl. In one embodiment, both R10 and R11 are (CH2)n such as CH2 (where n=1).
In one embodiment, at least one or both of R12 and R13 is hydrogen. In one embodiment, at least one or both of R12 and R13 is a C2-C8 substituted or unsubstituted alkenyl, such as propen-2-yl. In one embodiment, at least one or both of R12 and R13 is a C1-C8 substituted or unsubstituted alkyl, such as methyl or ethyl or any C1-C8 substituted or unsubstituted alkyl mentioned above in reference to any one of formulae (I), (II) or (III).
It is understood and clearly conveyed by this disclosure that each A, n, R10, R11, R12 and R13 disclosed in reference to formula (IV) intends and includes all combinations thereof the same as if each and every combination of A, n, R10, R11, R12 and R13 were specifically and individually listed.
Representative compounds of the formula (IV) include, e.g.:
In one embodiment, the polyamine is of the formula (V):
or a salt, solvate, or hydrate thereof, wherein n is an integer from 1 to 8; m is an integer from 1 to 8; R15 and R14 are independently selected from the group consisting of hydrogen, C1-C8 substituted or unsubstituted n-alkyl or (C3-C8) branched alkyl, C6-C20 substituted or unsubstituted aryl or heteroaryl and C7-C24 substituted or unsubstituted aralkyl or heteroaralkyl; R16 and R17 are independently hydrogen or a C1-C8 substituted or unsubstituted alkyl; and wherein the compound contains no more than three secondary amino groups except when R17 is a C1-C8 substituted or unsubstituted alkyl and wherein the compound is free from a methylphosphonate or hydroxy moiety.
In one embodiment, at least one or both of R15 and R14 is hydrogen. When only one of R15 and R14 is hydrogen, the R15 or R14 that is not hydrogen may be any other moiety listed above, such as a C6-C20 substituted or unsubstituted aryl or heteroaryl (e.g.; 4-isopropylbenzyl, 2-phenylbenzyl, 3,3,-diphenylpropyl and the like or any C6-C20 substituted or any unsubstituted aryl or heteroaryl listed above in reference to any one of formulae (I)-(IV)).
In one embodiment, at least one or both of R15 and R14 is a C1-C8 substituted or unsubstituted n-alkyl or (C3-C8) branched alkyl, such as methyl, ethyl, 3-methyl-butyl, 2-ethyl-butyl, 5-NH2-pent-1-yl, prop-1-yl-methyl(phenyl)phosphinate and the like or any C1-C8 substituted or unsubstituted n-alkyl or (C3-C8) branched alkyl listed above in reference to formulae (I)-(IV). In one embodiment, at least one or both of R15 and R14 is a C1-C8 substituted or unsubstituted n-alkyl, such as an n-alkyl substituted with a methyl(phenyl)phosphinate moiety or a NH2-substituted n-alkyl. In one embodiment, both R15 and R14 are C1-C8 substituted or unsubstituted n-alkyl or (C3-C8) branched alkyl moieties, such as when R15 and R14 are both 3-methyl-butyl or when R15 and R14 are both 2-ethyl-butyl. R15 and R14 may be different C1-C8 substituted or unsubstituted n-alkyl moieties, such as when one of R15 and R14 is propyl and the other is ethyl.
In one embodiment, at least one or both of R15 and R14 is a C7-C24 substituted or unsubstituted aralkyl or heteroaralkyl. In one embodiment, at least one or both of R15 and R14 is a C7-C24 substituted or unsubstituted aralkyl or heteroaralkyl having two rings, such as 2-phenylbenzyl, 4-phenylbenzyl, 2-benzylbenzyl, 3-benzylbenzyl, 3,3,-diphenylpropryl, 3-(benzoimidazolyl)-propyl and the like. In one embodiment, at least one or both of R15 and R14 is a C7-C24 substituted or unsubstituted aralkyl or heteroaralkyl having one ring, such as 4-isopropylbenzyl, 4-fluorobenzyl, 4-tert-butylbenzyl, 3-imidazolyl-propyl, 2-phenylethyl and the like. In one embodiment, one of R15 and R14 is a C7-C24 substituted or unsubstituted aralkyl or heteroaralkyl, such as any of the specific substituted or unsubstituted aralkyl or heteroaralkyl moieties listed for any other formula, and the other R15 and R14 is hydrogen or a C1-C8 substituted or unsubstituted n-alkyl or (C3-C8) branched alkyl, such as ethyl, methyl, 3-methylbutyl and the like.
For any compound of formula (V), m and n may be the same or different. In one embodiment, m does not equal n, such as when m is 1 and n is 2. For instance, in one embodiment, m is 1, n is 2 and both R15 and R14 are 2-benzylbenzyl. However, it is understood that all possible combinations of m, n, R15 and R14 are intended.
In one embodiment, at least one or both of R16 and R17 is hydrogen. In one embodiment, at least one or both of R16 and R17 is a C1-C8 substituted or unsubstituted alkyl, such as a methyl, ethyl and a C1-C8 alkyl substituted with e.g., an —NH—C1-C8 alkyl such as when at least one or both of R16 and R17 is —(CH2)3NHCH2CH3.
It is understood and clearly conveyed by this disclosure that each R14, R15, R16, R17, m, and n disclosed in reference to formula (V) intends and includes all combinations thereof the same as if each and every combination of R14, R15, R16, R17, m, and n were specifically and individually listed.
Representative compounds of the formula (V) include, e.g.:
In one embodiment, the polyamine is of the formula (VI):
or a salt, solvate, or hydrate thereof, wherein n is an integer from 1 to 12; m and p are independently an integer from 1 to 5; R18 and R19 are independently selected from the group consisting of hydrogen, C1-C8 unsubstituted alkyl (e.g., methyl, ethyl, Cert-butyl, isopropyl, pentyl, cyclobutyl), C1-C8 n-alkyl substituted with a cycloalkyl group comprising at least two rings, C7-C24 substituted or unsubstituted aralkyl or heteroaralkyl comprising at least two rings; and wherein: n is 1 when R18 and R19 are identical C1-C8 n-alkyl moieties substituted with a cycloalkyl group comprising at least two rings, or are identical aryl groups comprising at least two rings; and, at least one of R18 and R19 is either a C1-C8 n-alkyl substituted with a cycloalkyl group comprising at least two rings or a C7-C24 substituted or unsubstituted aralkyl comprising at least two rings.
In one embodiment, at least one or both of R18 and R19 is a C1-C8 n-alkyl substituted with a cycloalkyl group comprising at least two rings. The cycloalkyl group comprising at least two rings may be a spiro, fused or bridged cycloalkyl group. Representative examples of a C1-C8 n-alkyl substituted with a cycloalkyl group comprising two rings include moieties such as 2-(6,6-dimethylbicyclo[3.1.1]heptyl)ethyl and 2-(decahydronaphthyl)ethyl. In one embodiment, both R18 and R19 are 2-(6,6-dimethylbicyclo[3.1.1]heptyl)ethyl. In one embodiment, both R18 and R19 are 2-(decahydronaphthyl)ethyl. In one embodiment, one of R18 and R19 is 2-(6,6-dimethylbicyclo[3.1.1]heptyl)ethyl or 2-(decahydronaphthyl)ethyl and the other R18 and R19 is hydrogen or a C1-C8 unsubstituted alkyl such as ethyl.
In one embodiment, at least one or both of R18 and R19 is a C7-C24 substituted or unsubstituted aralkyl or heteroaralkyl comprising at least two rings, which rings may be but are not required to be fused. A substituted aralkyl or heteroaralkyl with reference to formula (VI) intends and includes alkanoyl moieties substituted with an aryl or heteroaryl group, i.e., —C(═O)-aryl, —C(═O)-aralkyl, —C (═O)-heteroaryl, and —C(═O)-heteroaralkyl. In one embodiment, the alkyl portion of the aralkyl or heteroaralkyl moiety is connected to the molecule via its alkyl moiety. For instance at least one or both of R18 and R19 may be an aralkyl moiety such as 2-phenylbenzyl, 4-phenylbenzyl, 3,3,-diphenylpropyl, 2-(2-phenylethyl)benzyl, 2-methyl-3-phenylbenzyl, 2-napthylethyl, 4-(pyrenyl)butyl, 2-(3-methylnapthyl)ethyl, 2-(1,2-dihydroacenaphth-4-yl)ethyl and the like. In another embodiment, at least one or both of R18 and R19 may be a heteroaralkyl moiety such as 3-(benzoimidazolyl)propanoyl, 1-(benzoimidazolyl)methanoyl, 2-(benzoimidazolyl)ethanoyl, 2-(benzoimidazolyl)ethyl and the like.
In one embodiment, each of m, n and p is the same, such as when m, n and p are each 1.
It is understood and clearly conveyed by this disclosure that each R18, R19, m, n and p disclosed in reference to formula (VI) intends and includes all combinations thereof the same as if each and every combination of R18, R19, m, n and p were specifically and individually listed.
Representative compounds of the formula (VI) include, e.g.:
In one embodiment, the polyamine is of the formula (VII):
or a salt, solvate, or hydrate thereof, wherein n is an integer from 1 to 12; m and p are independently an integer from 1 to 5; q is 0 or 1; R20 and R21 are independently selected from the group consisting of hydrogen, C1-C8 substituted or unsubstituted alkyl, —C(═O)—C1-C8 substituted or unsubstituted alkyl, —C(═O)—C1-C8 substituted or unsubstituted alkenyl, —C(═)—C1-C8 substituted or unsubstituted alkynyl, and C7-C24 substituted or unsubstituted aralkyl; and wherein the compound comprises at least one moiety selected from the group consisting of t-butyl, isopropyl, 2-ethylbutyl, 1-methylpropyl, 1-methylbutyl, 3-butenyl, isopent-2-enyl, 2-methylpropan-3-olyl, ethylthiyl, phenylthiyl, propynoyl, 1-methyl-1H-pyrrole-2-yl, trifluoromethyl, cyclopropanecarbaldehyde, halo-substituted phenyl, nitro-substituted phenyl, alkyl-substituted phenyl, 2,4,6-trimethylbenzyl, halo-S-substituted phenyl (such as para-(F3S)-phenyl, azido and 2-methylbutyl.
In one embodiment, q is 1. In one embodiment, q is 1 and n is 1.
In one embodiment at least one of R20 and R21 is hydrogen. In one embodiment at least one of R20 and R21 is C1-C8 substituted or unsubstituted alkyl, such as any of the substituted or unsubstituted alkyl moieties mentioned above for formulas (I)-(VI). In one embodiment at least one of R20 and R21 is a C7-C24 substituted or unsubstituted aralkyl, such as any of the C7-C24 substituted or unsubstituted aralkyl mentioned above for formulas (I)-(VI).
It is understood and clearly conveyed by this disclosure that each R20, R21, m, n, q and p disclosed in reference to formula (VII) intends and includes all combinations thereof the same as if each and every combination of R20, R21, m, n, q and p were specifically and individually listed.
Representative compounds of the formula (VII) include, e.g.:
In one embodiment, the polyamine is of the formula (VIII):
or a salt, solvate, or hydrate thereof, wherein m and p are independently an integer from 1 to 5; X is —(CH2)n- or cyclohex-1,3-diyl; n is an integer from 1 to 5; R22 and R23 are independently selected from the group consisting of hydrogen, n-butyl, ethyl, cyclohexylmethyl, cyclopentylmethyl, cyclopropylmethyl, cycloheptylmethyl, cyclohexyleth-2-yl, and benzyl; and when n is 5, at least one of R22 and R23 is hydrogen; when R22 is ethyl, R23 is hydrogen, n-butyl, cyclopentylmethyl, cyclohexyleth-2-yl or benzyl; and when R23 is ethyl, Rn is hydrogen, n-butyl, cyclopentylmethyl, cyclohexyleth-2-yl or benzyl; when X is cyclohex-1,3-diyl, R22 and R23 are not both benzyl or cyclopropylmethyl.
In one embodiment, X is —(CH2)n (e.g., CH2 where n is 1). In one embodiment, X is CH2 and m and p are both 1. In one embodiment, X is cyclohex-1,3-diyl. In one embodiment, X is cyclohex-1,3-diyl and m and p are both 1. In other embodiments, m and p are not the same, e.g., when m is 3 and p is 4.
It is understood and clearly conveyed by this disclosure that each R22, R23, m, n and p disclosed in reference to formula (VIII) intends and includes all combinations thereof the same as if each and every combination of R22, R23, m, n and p were specifically and individually listed.
Representative compounds of the formula (VIII) include, e.g.:
In one embodiment, the polyamine is of the formula (IX):
or a salt, solvate, or hydrate thereof, wherein p is an integer from 1 to 5; R24 is an amino-substituted cycloalkyl (e.g., a cycloalkyl group substituted with a primary, secondary, tertiary or quaternary amine) or a C2-C8 substituted or unsubstituted alkanoyl (which substituted alkanoyl may be substituted with one or more substituents such as those listed for “Substituted alkyl” including without limitation an alkanoyl substituted with a methyl and an alkylazide group); and R25 is a C1-C8 substituted or unsubstituted alkyl or a C7-C24 substituted or unsubstituted aralkyl, such as those listed above for any of formulae (I)-(VIII).
In one embodiment, R24 is an amino-substituted C3-C24 cycloalkyl, such as 5-NH2-cycloheptyl, 3-NH2-cyclopentyl and the like. In one embodiment, R25 is a C1-C8 substituted or unsubstituted alkyl, which includes an n-alkyl group substituted with a cycloalkyl, such as in cyclopropylmethyl. In one embodiment, R25 is cyclopropylmethyl or ethyl and R24 is 5-NH2-cycloheptyl or 3-NH2-cyclopentyl. In one embodiment, R24 is a C2-C8 substituted or unsubstituted alkanoyl and R24 is a C7-C24 substituted or unsubstituted aralkyl, such as 4-phenylbenzyl.
It is understood and clearly conveyed by this disclosure that each R24, R25 and p disclosed in reference to formula (IX) intends and includes all combinations thereof the same as if each and every combination of R24, R25 and p were specifically and individually listed.
Representative compounds of the formula (IX) include, e.g.:
For all formulae listed herein, such as formulae (I)-(IX), even if not explicitly stated, any substituent mentioned in one formula is intended to describe the same substituent in any other formula to the extent that the description conforms to the structural characterization of the formula described. For example, R1 in formula I is intended to describe any other R1 found in any other formula to the extent that the description conforms to the structural characterization of the formula described. Similarly, any description of, e.g., C1-C8 substituted or unsubstituted alkyl is intended to describe any other C1-C8 substituted or unsubstituted alkyl found in any other formula to the extent that the description conforms to the structural characterization of the formula described.
It is also recognized that any compounds listed as a particular salt thereof is not intended to limit the compound to such salt or form thereof. Similarly, where compounds are listed as a salt, the structure may or may not explicitly indicate positive or negative charges or the location thereof, and all possibilities thereof are intended. For instance, a compound listed as a 4HBr salt does not limit the compound to only the HBr salt and the compound may or may not show the + or − charges of the HBr salt, but rather all possibilities are intended.
Any of the polyamine compounds, such as compounds of the formula (I)-(IX) may be in a protected form, such as when any one or more amine (e.g., —NH—) is protected by a protecting group (Pg), such as in (—NPg-). Pg may be any protecting group, such as mesityl (e.g., NMes), Boc (e.g., —NBoc) or any other protecting group such as those described in, e.g. T. W. Green, P. G. M. Wuts, Protective Groups in Organic Synthesis, Wiley-Interscience, New York, 1999, which is incorporated herein by reference in its entirety.
Compounds within the scope of this invention and/or as described by any one or more of formulae (I)-(IX) include (but are not limited to) the compounds listed in Table A below.
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (M).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (I).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (II).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (III).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (IV).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (V).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (VI).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (VII).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (VIII).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds of formula (IX).
In another embodiment, the invention embraces a method of treating cancer, by administering a therapeutically effective amount of one or more of the compounds listed in Table A or Table B.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more compounds, where the compound has at least one guanidine moiety or at least one biguanide moiety, in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (M) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (I) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (II) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (III) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (IV) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (V) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (VI) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (VII) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (VIII) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds of formula (IX) in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
In another embodiment, the invention embraces a method of inhibiting a histone demethylase enzyme, such as LSD1, by contacting the enzyme with an amount of one or more of the compounds listed in Table A or Table B in an amount sufficient to inhibit the enzyme. The enzyme can be inhibited by at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 99%.
The disclosure includes all salts of the compounds described herein. The invention also includes all non-salt compounds of any salt of a compound named herein, as well as other salts of any salt of a compound named herein. In one embodiment, the salts of the compounds comprise pharmaceutically acceptable salts. Pharmaceutically acceptable salts are those salts which retain the biological activity of the free compounds and which can be administered as drugs or pharmaceuticals to humans and/or animals. The desired salt of a basic compound may be prepared by methods known to those of skill in the art by treating the compound with an acid. Examples of inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Examples of organic acids include, but are not limited to, formic acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, sulfonic acids, and salicylic acid. Salts of basic compounds with amino acids, such as aspartate salts and glutamate salts, can also be prepared. The desired salt of an acidic compound can be prepared by methods known to those of skill in the art by treating the compound with a base. Examples of inorganic salts of acid compounds include, but are not limited to, alkali metal and alkaline earth salts, such as sodium salts, potassium salts, magnesium salts, and calcium salts; ammonium salts; and aluminum salts. Examples of organic salts of acid compounds include; but are not limited to, procaine, dibenzylamine, N-ethylpiperidine, N,N′-dibenzylethylenediamine, and triethylamine salts. Salts of acidic compounds with amino acids, such as lysine salts, can also be prepared.
The disclosure includes all solvates of the compounds described herein, such as hydrates (in any ratios, e.g. monohydrates, dihydrates, hemihydrates, sesquihydrates), methanolates, ethanolates, etc.
Any compound described herein may occur in a combined salt and solvate form, for example the hyclate (monohydrochloride hemiethanolate hemihydrate) form.
The disclosure includes all stereoisomers of the compounds described herein, including diastereomers and enantiomers in optically pure or substantially optically pure form, as well as mixtures of stereoisomers in any ratio, including, but not limited to, racemic mixtures. Unless stereochemistry is explicitly indicated in a chemical structure or chemical name, the chemical structure or chemical name is intended to embrace all possible stereoisomers of the compound depicted.
The disclosure includes all crystal and non-crystalline forms of the compounds described herein, including all polymorphs, polycrystalline, and amorphous forms and any mixtures thereof.
The term “alkyl” refers to saturated aliphatic groups including straight-chain, branched-chain, cyclic groups, and combinations thereof, having the number of carbon atoms specified, or if no number is specified, having up to 12 carbon atoms. “Straight-chain alkyl” or “linear alkyl” groups refers to alkyl groups that are neither cyclic nor branched, commonly designated as “n-alkyl” groups. C1-C8 n-alkyl consists of the following groups: —CH2—, —CH2CH2—, —CH2CH2CH2—, —CH2CH2CH2CH2—, —CH2CH2CH2CH2CH2—, —CH2CH2CH2CH2CH2CH2—, —CH2CH2CH2CH2CH2CH2CH2—, and —CH2CH2CH2CH2CH2CH2CH2CH2—. Other examples of alkyl groups include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, butyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, n-pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, neopentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and adamantyl. Cycloalkyl groups can consist of one ring, including, but not limited to, groups such as cycloheptyl, or multiple bridged or fused rings, including, but not limited to, groups such as adamantyl or norbornyl groups. Cycloalkyl groups can also contain alkyl groups in addition to the cyclic portion, e.g., 2,6,6-trimethylbicyclo[3.1.1]heptane, 2-methyldecalin (2-methyldecahydronaphthalene), cyclopropylmethyl, cyclohexylmethyl, cycloheptylmethyl, and the like.
“Substituted alkyl” refers to alkyl groups substituted with one or more substituents including, but not limited to, groups such as halogen (including fluoro, chloro, bromo, and/or iodo-substituted alkyl such as a monohaloalkyl, dihaloalkyl, trihaloalkyl or multihaloalkyl, including a perhalooalkyl, for example, perfluoroalkyl, percholoralkyl, trifluoromethyl or pentachloroethyl), alkoxy, acyloxy, amino (including NH2, NHalkyl and N(alkyl)2), hydroxyl, mercapto, carboxy, benzyloxy, phenyl, benzyl, cyano, nitro, acyl, acylamino, amidino, alkyl amidino, thioamidino, aminoacyl, aryl, substituted aryl, aryloxy, azido, thioalkyl, —OS(O)2-alkyl, thioalkoxy, carboxaldehyde, carboalkoxy and carboxamide, or a functionality that can be suitably blocked, if necessary for purposes of the invention, with a protecting group. Examples of substituted alkyl groups include, but are not limited to, CF3, CF2CF3, and other perfluoro and perhalo groups; —CH2—OH; —CH2CH2CH(NH2)CH3, etc. Alkyl groups can be substituted with other alkyl groups, e.g., C3-C24 cycloalkyl groups.
The term “alkenyl” refers to unsaturated aliphatic groups including straight-chain (linear), branched-chain, cyclic groups, and combinations thereof, having the number of carbon atoms specified, or if no number is specified, having up to 12 carbon atoms, which contain at least one double bond (—C═C—). Examples of alkenyl groups include, but are not limited to, —CH2—CH═CH—CH3; and —CH2—CH2-cyclohexenyl, where the ethyl group can be attached to the cyclohexenyl moiety at any available carbon valence. The term “alkynyl” refers to unsaturated aliphatic groups including straight-chain (linear), branched-chain, cyclic groups, and combinations thereof, having the number of carbon atoms specified, or if no number is specified, having up to 12 carbon atoms, which contain at least one triple bond (—C≡C—). “Hydrocarbon chain” or “hydrocarbyl” refers to any combination of straight-chain, branched-chain, or cyclic alkyl, alkenyl, or alkynyl groups, and any combination thereof. “Substituted alkenyl,” “substituted alkynyl,” and “substituted hydrocarbon chain” or “substituted hydrocarbyl” refer to the respective group substituted with one or more substituents, including, but not limited to, groups such as halogen, alkoxy, acyloxy, amino, hydroxyl, mercapto, carboxy, benzyloxy, phenyl, benzyl, cyano, nitro, thioalkoxy, carboxaldehyde, carboalkoxy and carboxamide, or any group listed above for “Substituted alkyl,” or a functionality that can be suitably blocked, if necessary for purposes of the invention, with a protecting group.
“Aryl” or “Ar” refers to an aromatic carbocyclic group having a single ring (including, but not limited to, groups such as phenyl), two or more rings connected to each other (including, but not limited to, groups such as biphenyl and p-diphenylbenzene) or two or more condensed rings (including, but not limited to, groups such as naphthyl, anthryl, or pyrenyl), and includes both unsubstituted and substituted aryl groups. Aryls, unless otherwise specified, contain from 6 to 20 carbon atoms in the ring portion. A preferred range for aryls contains 6 to 12 carbon atoms in the ring portion. “Substituted aryls” refers to aryls substituted with one or more substituents, including, but not limited to, groups such as substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted hydrocarbon chains, halogen, alkoxy, acyloxy, amino, hydroxyl, mercapto, carboxy, benzyloxy, phenyl, benzyl, cyano, nitro, thioalkoxy, carboxaldehyde, carboalkoxy and carboxamide, or any group listed above for “Substituted alkyl,” or a functionality that can be suitably blocked, if necessary for purposes of the invention, with a protecting group. “Aralkyl” designates an alkyl-substituted aryl group, where any aryl can be attached to the alkyl; the alkyl portion can comprise one, two, or three straight chains of 1 to 6 carbon atoms each or one, two, or three branched chains of 3 to 6 carbon atoms each or any combination thereof. Aralkyl groups can consist of two aryl groups connected by an alkyl group, such as diphenylmethane or 2-methyl-1-(phenethyl)benzene. When an aralkyl group is indicated as a substituent, the aralkyl group can be connected to the remainder of the molecule at any available valence on either its alkyl moiety or aryl moiety; e.g., the tolyl aralkyl group can be connected to the remainder of the molecule by replacing any of the five hydrogens on the aromatic ring moiety with the remainder of the molecule, or by replacing one of the alpha-hydrogens on the methyl moiety with the remainder of the molecule. Preferably, the aralkyl group is connected to the remainder of the molecule via the alkyl moiety.
A preferred aryl group is phenyl, which can be substituted or unsubstituted. Substituents for substituted phenyl groups include lower alkyl (—C1-C4 alkyl), or a halogen (chlorine (Cl), bromine (Br), iodine (I), or fluorine (F); hydroxy (—OH), or lower alkoxy (—C1-C4 alkoxy), such as methoxy, ethoxy, propyloxy (propoxy) (either n-propoxy or i-propoxy), and butoxy (either n-butoxy, i-butoxy, sec-butoxy, or tert-butoxy); a preferred alkoxy substituent is methoxy. Substituted phenyl groups preferably have one or two substituents; more preferably, one substituent.
“Heteroalkyl,” “heteroalkenyl,” and “heteroalkynyl” refer to alkyl, alkenyl, and alkynyl groups, respectively, that contain the number of carbon atoms specified (or if no number is specified, having up to 12 carbon atoms) which contain one or more heteroatoms as part of the main, branched, or cyclic chains in the group. Heteroatoms include, but are not limited to, N, S, O, and P; N and O are preferred. Heteroalkyl, heteroalkenyl, and heteroalkynyl groups may be attached to the remainder of the molecule at any valence where a hydrogen can be removed, for example, at a heteroatom or at a carbon atom (if a valence is available at such an atom by removing a hydrogen). Examples of heteroalkyl groups include, but are not limited to, groups such as —O—CH3, —CH2—O—CH3, —CH2—CH2—O—CH3, —S—CH2—CH2—CH3, —CH2—CH(CH3)—S—CH3, —CH2—CH2—NH—CH2—CH2—, 1-ethyl-6-propylpiperidino, and morpholino. Examples of heteroalkenyl groups include, but are not limited to, groups such as —CH═CH—NH—CH(CH3)—CH2—. “Heteroaryl” or “HetAr” refers to an aromatic carbocyclic group having a single ring (including, but not limited to, examples such as pyridyl, imidazolyl, thiophene, or furyl) or two or more condensed rings (including, but not limited to, examples such as indolizinyl, indole, benzimidazole, benzotriazole, or benzothienyl) and having at least one hetero atom, including, but not limited to, heteroatoms such as N, O, P, or S, within the ring. Unless otherwise specified, heteroalkyl, heteroalkenyl, heteroalkynyl, and heteroaryl groups have between one and five heteroatoms and between one and twelve carbon atoms. “Substituted heteroalkyl,” “substituted heteroalkenyl,” “substituted heteroalkynyl,” and “substituted heteroaryl” groups refer to heteroalkyl, heteroalkenyl, heteroalkynyl, and heteroaryl groups substituted with one or more substituents, including, but not limited to, groups such as substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted benzyl, substituted or unsubstituted hydrocarbon chains, halogen, alkoxy, acyloxy, amino, hydroxyl, mercapto, carboxy, benzyloxy, phenyl, benzyl, cyano, nitro, thioalkoxy, carboxaldehyde, carboalkoxy and carboxamide, or any group listed above for “Substituted alkyl,” or a functionality that can be suitably blocked, if necessary for purposes of the invention, with a protecting group. Examples of such substituted heteroalkyl groups include, but are not limited to, piperazine, substituted at a nitrogen or carbon by a phenyl or benzyl group, and attached to the remainder of the molecule by any available valence on a carbon or nitrogen, —NH—SO2-phenyl, —NH—(C═O)O-alkyl, —NH—(C═O)O-alkyl-aryl, and —NH—(C═O)-alkyl. If chemically possible, the heteroatom(s) and/or the carbon atoms of the group can be substituted. A “heteroaralkyl” group is a heteroaryl group substituted with at least one alkyl group. The heteroatom(s) can also be in oxidized form, if chemically possible.
The term “alkoxy” as used herein refers to an alkyl, alkenyl, alkynyl, or hydrocarbon chain linked to an oxygen atom and having the number of carbon atoms specified, or if no number is specified, having up to 12 carbon atoms. Examples of alkoxy groups include, but are not limited to, groups such as methoxy, ethoxy, propyloxy (propoxy) (either n-propoxy or i-propoxy), and butoxy (either n-butoxy, i-butoxy, sec-butoxy, or tert-butoxy).
The terms “halo” and “halogen” as used herein refer to the Group VIIa elements (Group 17 elements in the 2005 IUPAC Periodic Table, IUPAC Nomenclature of Inorganic Chemistry) and include Cl, Br, F and I substituents.
“Protecting group” refers to a chemical group that exhibits the following characteristics: 1) reacts selectively with the desired functionality in good yield to give a protected substrate that is stable to the projected reactions for which protection is desired; 2) is selectively removable from the protected substrate to yield the desired functionality; and 3) is removable in good yield by reagents compatible with the other functional group(s) present or generated in such projected reactions. Examples of suitable protecting groups can be found in Greene et al, (1999) Protective Groups in Organic Synthesis, (Wiley-Interscience., New York). Amino protecting groups include, but are not limited to, mesitylenesulfonyl (Mts), benzyloxycarbonyl (CBz or Z), t-butyloxycarbonyl (Boc), t-butyldimethylsilyl (TBS or TBDMS), 9-fluorenylmethyloxycarbonyl (Fmoc), tosyl, benzenesulfonyl, 2-pyridyl sulfonyl, or suitable photolabile protecting groups such as 6-nitroveratryloxy carbonyl (Nvoc), nitropiperonyl, pyrenylmethoxycarbonyl, nitrobenzyl, dimethyl dimethoxybenzil, 5 bromo 7-nitroindolinyl, and the like. Hydroxyl protecting groups include, but are not limited to, Fmoc, TBS, photolabile protecting groups (such as nitroveratryl oxymethyl ether (Nvom)), Mom (methoxy methyl ether), and Mem (methoxy ethoxy methyl ether), NPEOC (4-nitrophenethyloxycarbonyl) and NPEOM (4 nitrophenethyloxymethyloxycarbonyl).
Specific Compounds
Examples of compounds useful in the invention are depicted in Table A. While the compounds are depicted as salts, such as the hydrobromide or trifluoroacetate salt, it is to be understood that the disclosure in the table embraces all salts, hydrates, and solvates of the compounds depicted therein, as well as the non-salt, non-hydrate/non-solvate form of the compound, as is well understood by the skilled artisan.
Synthetic Methods—Synthesis of Alkylpolyamines
Several synthetic methods are available for synthesis of polyamine analog compounds, including both symmetrically-substituted and asymmetrically-substituted polyamine analogs. Some of these methods are described in the following publications: Saab et al., J. Med. Chem. 36:2998 (1993); Bellevue et al., Bioorg. Med. Chem. Lett. 6:2765 (1996); Sirisoma et al., Tetrahedron Lett. 39:1489 (1998); Zou et al., Bioorg. Med. Chem. Lett. 11:1613 (2001), and Casero et al., J. Med. Chem. 44:1 (2001).
Scheme 1 illustrates a useful pathway to various polyamine analogs. The tetramesitylated intermediate 8 can be readily alkylated at both terminal nitrogens, since the hydrogens on these nitrogens are rendered acidic by the adjacent mesityl protecting group. Alkylation in the presence of 1.2 to 1.4 equivalents of alkyl halide or tosylate affords primarily the monosubstituted product 9, and disubstituted materials and unreacted starting material can then be separated and recycled (Bellevue et al., Bioorg. Med. Chem. Lett. 6:2765 (1996); Zou et al., Bioorg. Med. Chem. Lett. 11:1613 (2001)). The resulting monoalkylated derivative 9 can then be deprotected (30% HBr in AcOH), or realkylated with a different alkyl halide to provide the asymmetrically substituted intermediate 11. Deprotection of 11 then provides the desired asymmetrically substituted alkylpolyamine. Treatment of 8 with 2.2 equivalents of alkyl halide in the presence of NaH and DMF affords the bis-substituted intermediate 10, which upon deprotection yields the corresponding symmetrically substituted alkylpolyamine. Thus three distinct alkylpolyamines can be readily synthesized from a single intermediate, and the central carbon chain can be made in any desired length (n=0-8). Synthesis of the intermediate 8 is readily accomplished in large quantities using previously reported synthetic strategies (Bellevue et al., Bioorg. Med. Chem. Lett. 6:2765 (1996); Zou et al., Bioorg. Med. Chem. Lett. 11:1613 (2001)). A similar strategy can be used to access spermidine-like analogs of the form:
Other methods can be used for synthesis of the requisite polyamine backbone structures, which involve carbon nitrogen bond formation and selective nitrogen protection; some of these procedures are shown in Scheme 2.
Aminopropyl (or other aminoalkyl) moieties can be added to selectively protected primary amines such as 12 by standard peptide coupling techniques (Method A, Woster et al., J. Med. Chem. 32:1300 (1989)). Thus treatment of 12 with the protected beta-aminopropionate 13 (DCC, HoBt, N-methylmorpholine) affords the corresponding amide 14, which is then reduced in the presence of diborane (Woster et al., 1989) to afford the desired secondary amine 16. Compound 16 may be synthesized directly by reductive amination (Method B), in which the appropriate aldehyde 15 is added to 12 in the presence of sodium cyanoborohydride. Alkyl substituents that contain an allylic acetate functionality can also be appended to 12 using a palladium catalyzed coupling reaction that proceeds with retention of configuration (Method C, Sirisoma et al., Tetrahedron Lett. 39:1489 (1998)). This method can also be used to introduce phthalimide or benzylamine to an allylic acetate site as a synthetic equivalent for nitrogen. These nitrogens can then be deprotected and functionalized.
Synthetic Methods—Synthesis of Polyaminoguanidines
Synthesis of polyaminoguanidines can be carried out as outlined in Scheme 3. The requisite amine 19 (produced when necessary from the corresponding alkyl or aralkylcyanide) is reacted with cyanogen bromide (Goldin et al., U.S. Pat. No. 6,288,123 (2001)) to afford the corresponding aminocyanogen 20. When the desired amine is not commercially available, it can be prepared from the appropriate cyano compound by catalytic reduction (Bellevue et al., 1996, Zou et al., 2001). Intermediate 21 (Bellevue et al., 1996; Zou et al., 2001) is then coupled to 20 (chlorobenzene, reflux), followed by deprotection (30% Hbr in AcOH) to produce alkylpolyaminoguanidines of general structure 3. Using these methods, substituted polyaminoguanidine analogs (e.g., R═H, methyl, ethyl, cyclopropylmethylene, cycloheptylmethylene, phenyl, benzyl) can be synthesized. An analogous route (not shown) utilizing the N-Boc protection group was also employed.
Synthetic Methods—Synthesis of Polyaminobiguanides
The synthesis of polyaminobiguanides is described in Bi et al., Bioorg. Med. Chem. Lett. 16:3229 (2006), and is also outlined in Scheme 4.
A similar strategy is employed for the synthesis of alkylpolyaminobiguanides of general structure 4, as outlined in Scheme 4. Amines 23 (produced when necessary from the corresponding alkyl or aralkylcyanide) are converted to the corresponding cyanoguanidines 24 (NaN(CN)2, BuOH/H20) (Gerhard, R.; Heinz, B.; Herbert, F. J. Praktische Chem. (Leipzig), 1964, 26, 414-418), which were combined with 21 as previously described to afford the mesityl protected target molecules. Deprotection as described above then provided the substituted biguanides 4. An analogous route (not shown) utilizing the N-Boc protection group was also employed, as above.
Synthetic Methods—Solid Phase Synthesis
Solid phase synthetic techniques can be used for the rapid and efficient synthesis of both alkylpolyamines and their alpha-methyl homologs, as shown in Scheme 4. Compound 22 can be produced using a commercially available trityl chloride resin, as described in Wang et al., J. Am. Chem. Soc., 95(4):1328 (1973), where the attached amine is primary or secondary prior to attachment, an alpha-methyl is present or absent, and the X group is either a protected amine or a synthetic equivalent such as an azide or a phthalamide. This intermediate is then deprotected or converted to the corresponding primary amine 23. Three strategies can be used for chain elongation: 1. reductive amination with aldehydes 24 in the presence of sodium cyanoborohydride to produce 25; 2. addition of an appropriate carboxylate 26 under peptide coupling conditions (Woster et al., J. Med. Chem. 32:1300 (1989)), followed by diborane reduction of the resulting amide, yielding 27; 3. direct alkylation with a protected halide (Woster et al., J. Med. Chem. 32:1300 (1989)) such as 28, to afford intermediates 29. Repetition of these steps then allows the synthesis of a variety of alkylpolyamines and alpha-methyl-alkylpolyamines with substituents as desired.
Biological Applications—Lysine-Specific Demethylase-1 (LSD1) Inhibitors
Histones are proteins found in eukaryotic cells which act as support scaffolds for DNA (sometimes compared to a protein spool supporting the DNA thread). Histones, together with other proteins and DNA, form the chromatin of the cell nucleus. Because of their close association with DNA, histones play a role in gene regulation. The tails of histone proteins are a frequent site for covalent modifications which affect gene expression.
The enzyme lysine-specific demethylase-1 (LSD1; also known as BHC110 and KIAA0601) is an enzyme that affects the covalent modification of histone tails, by demethylating lysine 4 of the histone H3. Shi et al. (Cell, 119:941 (2004)) showed that RNAi inhibition of LSD1 led to an increase in H3 lysine 4 methylation, followed by de-repression of the target genes. Thus LSD1 apparently represses transcription by demethylating histone H3. Conversely, inhibition of LSD1 allows transcription by preventing demethylation.
Because of the observed homology between the active site of LSD1 and monoamine oxidase (MAO), Lee et al. (Chemistry & Biology 13:563 (2006)) tested various MAO inhibitors for their ability to inhibit LSD1. They identified tranylcypromine ((1R,2S)-2-phenylcyclopropan-1-amine) as an inhibitor with an IC50 less than 2 micromolar. Treating P19 embryonal carcinoma cells with tranylcypromine led to transcriptional de-repression of the Egr1 and Oct4 genes.
International Patent Application WO 2006/071608 is directed to a method for monitoring eukaryotic histone demethylase activity, methods for up-regulating and down-regulating methylated histone-activated genes, and a method for treating or preventing a disease (e.g., a hyperproliferative disease such as cancer) by modulating the level of protein or the activity of a histone demethylase.
In view of the importance of gene regulation, and the ability to affect gene regulation by inhibiting or modulating LSD1, inhibitors of the enzyme may have significant therapeutic potential. Table B shows compounds tested for LSD1 inhibitory activity While the compounds are depicted as free bases, it is to be understood that the disclosure in the table embraces all salts, hydrates, and solvates of the compounds depicted therein, as well as the non-salt, non-hydrate/non-solvate form of the compound, as is well understood by the skilled artisan. Several of the polyamine, polyamine/guanidine, and polyamine/biguanide compounds disclosed herein have activity as LSD1 inhibitors.
Biological Applications—Treatment of Cancer
Several polyamine compounds and polyamine analogs have displayed potent anticancer activity. It is believed that polyamines and polyamine analogs enter cells via the polyamine transport system and down-regulate the polyamine biosynthetic enzymes ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMet-DC). The antitumor activity of the bis(ethyl) polyamine analogs is thought to be due to their ability to superinduce spermidine/spermine-N1-acetyltransferase (SSAT), the rate-limiting step in the polyamine back-conversion pathway. Subsequent polyamine oxidase (PAO)-mediated oxidation of the resulting acetylated polyamines then produces hydrogen peroxide which ultimately initiates the cell death program. Studies have revealed analogs that inhibit tumor cell growth through induction of SSAT, by initiating apoptosis in the presence and absence of SSAT induction, and by interference with tubulin depolymerization. Recent data suggests that human polyamine oxidase exists in two distinct forms, and that oxidation of polyamine analogues by mammalian spermidine oxidase (SMO(PAOh1) may play a role in the antitumor effects of some analogs. This hypothesis is supported by the facts that the alkylpolyamine analogues N1-ethyl-N11-[(cycloheptyl)methy]-4,8-diazaundecane (CHENSpm) is detoxified by polyamine oxidase, and that the antimicrosporidial analogue BW-1 (N,N′-bis[3-[([1,1′-biphenyl]-2-ylmethyl)amino]propyl]-1,7-heptanediamine) is substrate for the polyamine oxidase of Encephalitoozoon cuniculi. It is now evident that alkylpolyamines can effect tumor cell growth by a variety of known and unknown pathways.
“Treating” or “to treat” a disease using the methods of the invention is defined as administering one or more polyamines or polyamine analogs, with or without additional therapeutic agents, in order to palliate, ameliorate, stabilize, reverse, slow, delay, prevent, reduce, or eliminate either the disease or the symptoms of the disease, or to retard or stop the progression of the disease or of symptoms of the disease. “Therapeutic use” of the polyamines and polyamine analogs is defined as using one or more polyamines or polyamine analogs to treat a disease (including to prevent a disease), as defined above. A “therapeutically effective amount” is an amount sufficient to treat (including to prevent) a disease, as defined above. Prevention or suppression can be partial or total.
The compounds disclosed herein have anticancer activity, which has been demonstrated in a variety of human tumor cell types representing the major forms of lung, breast, prostate, and colon cancers. Thus the compounds disclosed herein can be used to treat cancer, including lung cancer, breast cancer, prostate cancer, and colon cancer, or to prevent cancer, including prevention of lung cancer, breast cancer, prostate cancer, and colon cancer.
Experimental Results and Protocols
MTS dose response experiments in H157, H82, and A549 cells following a 96 hr exposure with select compounds were performed. MTS is a standard colorimetric assay used for measuring metabolic activity in cells. MTS experiments were performed by CellTiter 96® AQueuos One Solution Cell Proliferation Assay from Promega Corporation. Briefly, the cells were seeded at 3000 cells/well on a 96 well tissue culture plate containing 100 ul of medium/well and allowed to attach overnight. The medium was then aspirated and replaced with 100 ul of fresh medium containing the appropriate concentration of the compound being tested and incubated for 96 hrs at 37° C. and 5% CO2. Compounds are routinely tested at concentrations ranging from 0.1 micromolar to 50 micromolar. Wells not containing the test compound were present and used as a control. Following treatment, 20 ul of MTS reagent was added to each well and incubated at 37° C. for 1.5 hrs. The absorbance of each well was then measured at 490 nm and used to determine the metabolic activity of the cells in the presence of the test compound, relative to the control. IC50 values for the test compounds were extracted based on the results.
The results for H157, H82, and A549 cells are shown in Tables 2, 3 and 4 respectively. Note that the results for a 72 hr exposure in addition to the 96 hr exposure are shown for compound 49-TDW-29C in the H157 cells. The first column contains the compound identifier and the second column contains the IC50 values (when a range is shown, e.g., 1-10 uM, this indicates that the IC50 lies somewhere between the two endpoints of the range; the endpoints are the concentrations actually tested, one of which is lower than the IC50 and one of which is higher).
MTT dose response experiments in 235, MCF7, 435, and 10A cells were performed. MTT is a standard colorimetric assay used for measuring metabolic activity in cells. Briefly, about 200 ul of media not containing cells was added to column A of a 96 well plate and used as a blank. Next, 200 ul of media containing cells was added to the remaining wells and incubated overnight. The remaining wells contain about 4000-5000 MCF7 cells/well, 3000 231 cells/wells, 12,000 468 cells/well, or 9000 MCF 10A cells/well. Following incubation, the media in the wells was aspirated and replaced with 200 ul of fresh media in columns A and B of the 96 well plate. Column B was used as a control. Next 200 ul of fresh media containing the compound being tested was added to the remaining wells and incubated for 96 hrs. Compounds are routinely tested at concentrations ranging from 0.1 micromolar to 50 micromolar. Following incubation for 96 hrs, the media in each well was aspirated and replaced with 100 ul of 5 mg/ml MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution in Serum-Free media and incubated for 4 hours. Following incubation with MTT solution, the MTT solution was removed from the wells and replaced with 200 ul of a 1:1 Etoh+DMSO solution and incubated for 20 minutes. Following incubation with the Etoh+DMSO solution the plates were read at 540 nm and used to determine the metabolic activity of the cells in the presence of the test compound, relative to the control. IC50 values for the test compounds were extracted based on the results.
The results of an MTT assay after 96 hrs of treatment with compound 39-TDW-47c at different concentrations in 231, MCF7, 435, and 10A cells is shown in
SSAT (spermidine/spermine-NI-acetyltransferase) activity experiments in H157, H82, and A549 cells following exposure to select compounds were performed. A detailed protocol for determining SSAT activity is described in Casero et al., Cancer Research, 49:3829 (1989). Briefly, the SSAT activity was measured by harvesting the treated cells at the exposure time. The cells were then lysed and treated with spermidine, and 1-[14C]acetyl coenzyme A for 5 minutes. Enzyme activity was measured in term of picomoles of [14C]acetylspermidine formed per mg of cell protein per min (pmol/mgP/min).
The results are show in tables 5 (H157), 9 (H82), and 12 (A549) respectively. In Tables 5 and 12, the compound identifier, treatment concentration, control activity, SSAT activity following exposure and exposure time are listed in columns 1, 2, 3, 4, and 5 respectively. The activity in Tables 5 and 12 is reported as picomoles of SSAT per mg of protein per min. Table 9 lists the compound identifier, the exposure concentration, the activity, and exposure time in columns 1, 2, 3 and 4 respectively. No SSAT induction was observed for H82 cells and thus the values of the control and activity following exposure are not listed.
Putrescine, spermidine, and spermine polyamine levels in H157 and H82 cells following exposure to select compounds were performed. Polyamine levels were determined using the precolumn dansylation labeling, reverse-phase high-pressure liquid chromatography method as described by Kabra et al., J. Chromotography, 380:19 (1986). The results are show in Tables 6 and 11 for H157 and H82 cells respectively. The compound identifier, treatment concentration, observed polyamine level, and exposure time are listed in columns 1, 2, 3, and 4 respectively. Polyamine levels are reported as increased (inc), decrease (dec), or no change (N/C). In some case the specific levels of putrescine, spermidine, and/or spermine are listed.
SMO (Spermine Oxidase) activity in H157 cells following treatment with compound 46-TDW-34C is shown in Table 7, A detailed protocol for measuring SMO activity is described in Wang et al., Cancer Research, 61:5370 (2001). The compound identifier, the treatment concentration, the control activity, the activity following treatment and the exposure time are listed in columns 1, 2, 3, 4, and 5 respectively. The activity results are reported in picomoles of spermine converted per mg of cell protein per min (pmol/mgP/min).
ODC (Ornithine decarboxylase) activity experiments in H157 were performed. A detailed protocol for measuring ODC activity is described in Pegg et al., Methods Enzymology, 94:158 (1983). The results are show in Table 10. The compound identifier, treatment concentration, control activity, activity following treatment, and exposure time are listed in columns 1, 2, 3, 4 and 5 respectively. The activity results are reported in picomoles of CO2 released per mg of cell protein per hour (pmol/mgP/hr).
Treatment induced cell cycle measurements in H157 cells were performed. Following exposure of the cells to a compound of interest, at a concentration of 10 uM, for 24 hrs, the cells were harvested, prepared and transferred to a FACS for cell cycle analysis. (See Carlisle et al., Clinical Cancer Research 8:2684 (2002) and references therein.) The results are shown in Table 8. The results depict the percentage of cells which are in the G1 phase, S phase, and G2/M phases.
The disclosures of all publications, patents, patent applications and published patent applications referred to herein by an identifying citation are hereby incorporated herein by reference in their entirety.
Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is apparent to those skilled in the art that certain minor changes and modifications will be practiced. Therefore, the description and examples should not be construed as limiting the scope of the invention.
This application claims priority benefit of U.S. Provisional Patent Application No. 60/707,420, filed Aug. 10, 2005. The entire contents of that application are hereby incorporated by reference herein.
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| Number | Date | Country | |
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| 20100273745 A1 | Oct 2010 | US |
| Number | Date | Country | |
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| 60707420 | Aug 2005 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 11463840 | Aug 2006 | US |
| Child | 12763722 | US |
