Compounds Having Anti-Adhesive Effects on Cancer Cells

Abstract

Compounds of the form O-(x-L)n, where O is an oligonucleoside having at least a plurality of N3′->P5′ thiophosphoramidate (NPS) internucleoside linkages, a conjugated lipid moiety L, and at least one G-rich sequence motif as described, are effective to morphologically alter and reduce adhesion of cancer cells.

1 . A compound having a structure represented by O-(x-L)n, where
 (a) O is a polynucleoside moiety comprising a sequence of nucleosides and linkage moieties, wherein   (i) at least 50% of said linkage moieties are selected from: 3′-NH—P(O)(S − )-5′; 3′-NH—P(O)(S—)—{OR} m —Y—P(O)(S − )-5′; and 3′-Y—R—O—P(O)(S − )-5′; where Y is NH or O; R is a stable linear chain two to six atoms in length having bonds selected from alkyl, alkenyl, ether, thioether, and amino; and in is 1 to 3; and   (ii) said polynucleoside moiety includes at least one motif selected from GGG and GG(W) 1-3 GG, containing residues G and/or W, where G is guanosine and W is a nucleoside or the moiety —OR—, where R is as defined above, and the inter-residue linkages within said motif are N3′→P5′ thiophosphoramidate (3′-NH—P(O)(S+5′) or phosphorothioate (3′-O—P(O)(S—)-5′) linkages;   (b) x is an optional linker group,   (c) L is a lipid moiety, and   (d) n is 1 or 2;   with the proviso that, when said polynucleoside moiety O includes a sequence seven or more nucleobases in length that is complementary to a region of hTR (SEQ ID NO: 1), the compound O-(x-L)n is not a telomerase inhibitor.   
     
     
         2 . The compound of  claim 1 , wherein the compound is not a telomerase inhibitor. 
     
     
         3 . The compound of  claim 1 , wherein Y is NH. 
     
     
         4 . The compound of  claim 1 , wherein m is 1. 
     
     
         5 . The compound of  claim 1 , wherein at least 75% of said linkage moieties are selected from 3′-NH—P(O)(S − )-5′ (NPS) and 3′-NH—P(O)(S—)-{OR} m —Y—P(O)(S − )-5′, where m is 1. 
     
     
         6 . The compound of  claim 1 , wherein said inter-residue linkages are 3′-NH—P(O)(S − )-5′ (NPS) linkages. 
     
     
         7 . The compound of  claim 1 , wherein at least 50% of internucleoside linkages are 3′-NH—P(O)(S—)-5′ (NPS) linkages. 
     
     
         8 . The compound of  claim 1 , wherein R is a stable linear chain three to five atoms in length having bonds selected from alkyl and ether linkages. 
     
     
         9 . The compound of  claim 1 , wherein R is —(CH 2 ) n —, where n is 3 to 5. 
     
     
         10 . The compound of  claim 1 , wherein the sum of nucleosides and groups —OR— in the polynucleoside moiety O is from 5 to about 30. 
     
     
         11 . The compound of  claim 10 , wherein the sum of nucleosides and groups —OR— in the polynucleoside moiety O is from 7 to about 15. 
     
     
         12 . The compound of  claim 1 , wherein the ratio of nucleosides to groups —OR— in the polynucleoside moiety O is 1:1 or greater. 
     
     
         13 . The compound of  claim 1 , wherein O contains a sequence motif of at least three consecutive guanosine (G) nucleosides directly linked by NPS linkages. 
     
     
         14 . The compound of  claim 1 , wherein O contains a sequence motif GG(W) 1-3 GG, where G is guanosine and W is a nucleoside, and the linkages within said motif are N3′→P5′ thiophosphoramidate (NPS) linkages. 
     
     
         15 . The compound of  claim 1 , wherein the lipid L comprises a linear hydrocarbon moiety at least 12 carbon atoms in length, and is attached to the 3′ or 5′ terminus of said oligonucleoside. 
     
     
         16 . The compound of  claim 15 , wherein the lipid L is a C12 to C24 linear hydrocarbon. 
     
     
         17 . The compound of  claim 10 , wherein the lipid L is a saturated or monounsaturated hydrocarbon. 
     
     
         18 . The compound of  claim 15 , wherein the lipid L is a palmitic or oleic acid derivative and is attached to said terminus via a glycerol or aminoglycerol linker. 
     
     
         19 . The compound of  claim 1 , containing at least one linkage 3′-NH—P(O)(S—)—O—R—Y—P(O)(S − )-5′, where Y is NH or O, and R is a stable linear chain two to six atoms in length having bonds selected from alkyl, alkenyl, ether, thioether, and amino. 
     
     
         20 . The compound of  claim 19 , where R is —(CH 2 ) 3 —. 
     
     
         21 . A compound in accordance with  claim 20 , wherein said compound is designated herein as GRN163L A/Link. 
     
     
         22 . A method of inhibiting cell adhesion, comprising
 contacting said cells with a compound having a structure represented by O-(x-L)n, where   (a) O is a polynucleoside moiety comprising a sequence of nucleosides and linkage moieties, wherein   (i) at least 50% of said linkage moieties are selected from: 3′-NH—P(O)(S − )-5′; 3′-NH—P(O)(S—)—{OR} m —Y—P(O)(S − )-5′; and 3′-Y—R—O—P(O)(S − )-5′, where Y is NH or O;   R is a stable linear chain two to six atoms in length having bonds selected from alkyl, alkenyl, ether, thioether, and amino; and m is 1 to 3; and   (ii) said sequence includes at least one motif selected from GGG and GG(W) 1-3 GG, containing residues G and/or W, where G is guanosine and W is a nucleoside or the moiety —O—R—, where R is as defined above, and the inter-residue linkages within said motif are N3′→P5′ thiophosphoramidate (3′-NH—P(O)(S—)-5′) or phosphorothioate (3′-O—P(O)(S—)-5′) linkages;   (b) x is an optional linker group,   (c) L is a lipid moiety, and   (d) n is 1 or 2;   with the proviso that, when said polynucleoside moiety O includes a sequence seven or more nucleobases in length that is complementary to a region of hTR (SEQ ID NO: 1), the compound O-(x-L)n is not a telomerase inhibitor.   
     
     
         23 . The method of  claim 22 , wherein the compound is not a telomerase inhibitor. 
     
     
         24 . The method of  claim 22 , wherein said cells are cancer cells. 
     
     
         25 . A method of treating cancer in a patient, by inhibiting adhesion of cancer cells, comprising administering to the patient an effective amount of a compound having a structure represented by O-(x-L)n, where
 (a) O is a polynucleoside moiety comprising a sequence of nucleosides and linkage moieties, wherein   (i) at least 50% of said linkage moieties are selected from: 3′-NH—P(O)(S − )-5′; 3′-NH—P(O)(S—)—{OR} m —Y—P(O)(S − )-5′; and 3′-Y—R—O—P(O)(S − )-5′, where Y is NH or O; R is a stable linear chain two to six atoms in length having bonds selected from alkyl, alkenyl, ether, thioether, and amino; and m is 1 to 3; and   (ii) said sequence includes at least one motif selected from GGG and GG(W) 1-3 GG, containing residues G and/or W, where G is guanosine and W is a nucleoside or the moiety —O—R—, where R is as defined above, and the inter-residue linkages within said motif are N3′→P5′ thiophosphoramidate (3′-NH—P(O)(S—)-5′) or phosphorothioate (3′-O—P(O)(S—)-5′) linkages;   (b) x is an optional linker group,   (c) L is a lipid moiety, and   (d) n is 1 or 2;   with the proviso that, when said polynucleoside moiety O includes a sequence seven or more nucleobases in length that is complementary to a region of hTR (SEQ ID NO: 1), the compound O-(x-L)n is not a telomerase inhibitor.   
     
     
         26 . The method of  claim 25 , wherein the compound is not a telomerase inhibitor. 
     
     
         27 . The method of  claim 25 , wherein said administering accompanies tumor biopsy or tumor-reductive surgery. 
     
     
         28 . A compound having a structure represented by O-(x-L)n, where
 (a) O is a polynucleoside moiety comprising a sequence of nucleosides and linkage moieties, wherein   (i) at least 50% of said linkage moieties are selected from: 3′-NH—P(O)(S)-5′; 3′-NH—P(O)(S—)—{OR} m —Y—P(O)(S − )-5′; and 3′-Y—R—O—P(O)(S − )-5′, where Y is NH or O; R is a stable linear chain two to six atoms in length having bonds selected from alkyl, alkenyl, ether, thioether, and amino; and m is 1 to 3; and   (ii) said sequence includes at least one motif selected from GGG and GG(W) 1-3 GG, containing residues G and/or W, where G is guanosine and W is a nucleoside or the moiety —O—R—, where R is as defined above, and the inter-residue linkages within said motif are N3′→P5′ thiophosphoramidate (3′-NH—P(O)(S—)-5′) or phosphorothioate (3′-O—P(O)(S—)-5′) linkages;   (b) x is an optional linker group,   (c) L is a lipid moiety, and   (d) n is 1 or 2;   with the proviso that, when said polynucleoside moiety O includes a sequence seven or more nucleobases in length that is complementary to a region of hTR (SEQ ID NO: 1), the compound O-(x-L)n is not a telomerase inhibitor;   for use in medicine.   
     
     
         29 . The compound of  claim 28 , wherein the compound is not a telomerase inhibitor. 
     
     
         30 . Use of a compound having a structure represented by O-(x-L)n, where
 (a) O is a polynucleoside moiety comprising a sequence of nucleosides and linkage moieties, wherein   (i) at least 50% of said linkage moieties are selected from: 3′-NH—P(O)(S − )-5′; 3′-NH—P(O)(S—)—{OR} m —Y—P(O)(S − )-5′; and 3′-Y—R—O—P(O)(S − )-5′, where Y is NH or O; R is a stable linear chain two to six atoms in length having bonds selected from alkyl, alkenyl, ether, thioether, and amino; and m is 1 to 3; and   (ii) said sequence includes at least one motif selected from GGG and GG(W) 1-3 GG, containing residues G and/or W, where G is guanosine and W is a nucleoside or the moiety —O—R—, where R is as defined above, and the inter-residue linkages within said motif are N3→P5′ thiophosphoramidate (3′-NH—P(O)(S—)-5′) or phosphorothioate (3′-O—P(O)(S—)-5′) linkages;   (b) x is an optional linker group,   (c) L is a lipid moiety, and   (d) n is 1 or 2;   with the proviso that, when said polynucleoside moiety 0 includes a sequence seven or more nucleobases in length that is complementary to a region of hTR (SEQ ID NO: 1), the compound O-(x-L)n is not a telomerase inhibitor;   for the manufacture of a medicament for treating cancer.   
     
     
         31 . The use of  claim 30 , wherein the compound is not a telomerase inhibitor. 
     
     
         32 . The compound of  claim 1 , wherein the oligonucleotide comprises the sequence AACGTTGAGGGGCAT (SEQ ID NO:6), wherein all of said linkage moieties are NPS linkages, and the lipid is a 5′-palmitoyl group. 
     
     
         33 . The compound of  claim 1 , wherein the oligonucleotide comprises the sequence AACGAGTTGGGGCAT (SEQ ID NO:7), wherein all of said linkage moieties are NPS linkages, and the lipid is a 5′-palmitoyl group.