Lipids, lipid compositions, and methods of using them

Abstract

Disclosed are formulation and optimization protocols for delivery of therapeutically effective amounts of biologically active agents to liver, tumors, and/or other cells or tissues. Also provided are compositions and uses for cationic lipid compounds of formula (I). The invention also relates to compositions and uses for stealth lipids of formula (XI). Also provided are processes for making such compounds, compositions, and formulations, plus methods and uses of such compounds, compositions, and formulations to deliver biologically active agents to cells and/or tissues.

1 . A composition comprising at least one cationic lipid, at least one helper lipid and at least one stealth lipid for delivery of a biologically active agent, wherein the biologically active agent is for delivery to a tissue or cell selected from:
 a) a liver or liver cells, wherein the composition has a cationic lipid with a pKa of from about 6.2 or above;   b) a tumor or tumor cell, wherein the composition has a cationic lipid with a pKa of from about 6.2 or below;   c) a liver or liver cells, wherein the composition has a cationic lipid with a pKa of from about 5.1 to about 7.4; and   d) a tumor or tumor cell, wherein the composition has a cationic lipid with a pKa of from about 5.0 to about 6.7.   
     
     
         2 . A compound of formula (I): 
       
         
           
           
               
               
           
         
       
       or a salt or pharmaceutically acceptable derivative thereof, 
       wherein:
 R 1  and R 2  together with the nitrogen atom to which they are attached form an optionally substituted C 3-20 -heterocycloalkyl, C 3-20 -heterocycloalkenyl, C 3-20 -heterocycloalkynyl or C 5-20 -heteroaryl group; 
 a is absent or optionally substituted C 1-4  alkylene; 
 b is absent or optionally substituted C 1-4  alkylene; 
 c is absent or optionally substituted C 1-4  alkylene; 
 X 1  is O or S; 
 X 2  is O or S; 
 Y 1  is optionally substituted C 10-30 alkenyl, C 10-30 alkynyl, C 10-30 heteroalkenyl or C 10-30 heteroalkynyl; 
 L is absent or -(L b ) d -(L b ) e -(L c ) f -, wherein
 L a  is optionally substituted C 1-15 alkylene, C 1-15  alkenylene, C 1-15 alkynylene, C 1-15 heteroalkylene, C 1-15 heteroalkenylene or C 1-15 heteroalkynylene; 
 L b  is optionally substituted C 6-14 arylene or C 5-13 heteroarylene; 
 L c  is optionally substituted C 1-15 alkylene, C 1-15 alkenylene, C 1-15 alkynylene, C 1-15 heteroalkylene, C 1-15 heteroalkenylene or C 1-15 heteroalkynylene; 
 d is 0 or 1; 
 e is 0 or 1; and 
 f is 0 or 1; and 
 
 Y 2  is an optionally substituted steroid. 
 
     
     
         3 . The compound of  claim 2 , wherein one or more substituents are selected from one or more the following:
 (a) a is selected from optionally substituted C 1-2  alkylene and optionally substituted C 1  alkylene;   (b) b is selected from optionally substituted C 0-2  alkylene and optionally substituted C 1  alkylene;   (c) c is absent or is optionally substituted C 1  alkylene; and/or   (d) a, b and c are unsubstituted.   (e) R 1  and R 2  together with the nitrogen atom to which they are attached form an optionally substituted C 3-20 -heterocycloalkyl, C 3-20 -heterocycloalkenyl or C 3-20 -heterocycloalkynyl group;   (f) wherein R 1  and R 2  together with the nitrogen atom to which they are attached form a compound selected from a cyclic optionally substituted C 5-16  group and a cyclic optionally substituted C 5-12  group;   (g) R 1  and R 2  together with the nitrogen atom to which they are attached form a cyclic optionally substituted C 5  group, C 6  group or C 7  group; and   (h) R 1  and R 2  together with the nitrogen atom to which they are attached are selected from at least one of the head groups H 1  to H 52 .   (i) wherein X 1  is O;   (j) wherein X 2  is O;   (k) wherein both X 1  and X 2  is O;   (l) wherein L comprises at least one heteroatom;   (m) wherein L comprises at least one O atom;   (n) wherein L c  is selected from one of formulae L c-i  to L c-xxxxiii .   (o) wherein d is 0; e is 0, and f is 1;   (p) wherein Y 1  is a C 12-25  group;   (q) wherein Y 1  has at least one alkene group;   (r) wherein Y 1  has at least one cis unsaturated alkene group;   (s) wherein Y 1  is selected from Y 1-i  to Y 1-vii ;   (t) wherein Y 2  is linked to L via an oxygen atom on the optionally substituted steroid;   (u) wherein Y 2  is a sterol in which the hydrogen atom of the hydroxy group at the 3-position of the A steroid ring has been removed;   (v) wherein Y 2  is the sterol of part (u), and the sterol is selected from the group consisting of: annasterol; avenasterol; beta-sitosterol; brassicasterol; calciferol; campesterol; chalinosterol; chinasterol; cholestanol; cholesterol; coprostanol; cycloartenol; dehydrocholesterol; desmosterol; dihydrocalciferol; dihydrocholesterol; dihydroergosterol; dinosterol; epicholesterol; ergosterol; fucosterol; hexahydrolumisterol; hexaol; hydroxycholesterol; lanosterol; lumisterol; parkeol; poriferasterol; saringosterol; sitostanol; sitosterol; stigmastanol; stigmasterol; weinbersterol; zymosterol; sterol bile acids (including one or more selected from cholic acid; chenodeoxycholic acid; glycocholic acid; taurocholic acid; deoxycholic acid, and lithocholic acid); and/or a salt or a pharmaceutically acceptable derivative thereof; and   (w) wherein Y 2  is a cholesterol.   
     
     
         4 . A stealth lipid of formula (XI) 
       
         
           
           
               
               
           
         
       
       or a salt or pharmaceutically acceptable derivative thereof, 
       wherein
 Z is a hydrophilic head group component selected from PEG and polymers based on poly(oxazoline), poly(ethylene oxide), poly(vinyl alcohol), poly(glycerol), poly(N-vinylpyrrolidone), poly[N-(2-hydroxypropyl)methacrylamide] and poly(amino acid)s, wherein the polymer may be linear or branched, and wherein the polymer may be optionally substituted; 
 wherein Z is polymerized by n subunits; 
 n is a number-averaged degree of polymerization between 10 and 200 units of Z, wherein n is optimized for different polymer types; 
 L 1  is an optionally substituted C 1-10  alkylene or C 1-10 heteroalkylene linker including zero, one, two or more of an ether (e.g., —O—), ester (e.g., —C(O)O—), succinate (e.g., —O(O)C—CH 2 —CH 2 —C(O)O—)), carbamate (e.g., —OC(O)—NR′—), carbonate (e.g., —OC(O)O—), ketone (e.g., —C—C(O)—C—), carbonyl (e.g., —C(O)—), urea (e.g., —NRC(O)NR′—), amine (e.g., —NR′—), amide (e.g., —C(O)NR′—), imine (e.g., —C(NR′)—), thioether (e.g., —S—), xanthate (e.g., —OC(S)S—), and phosphodiester (e.g., —OP(O) 2 O—); any of which may be substituted by zero, one or more Z groups; 
 wherein R′ is independently selected from H, —NH—, —O—, —S—, a phosphate or an optionally substituted C 1-10  alkylene; 
 X 1  and X 2  are independently selected from a carbon or a heteroatom selected from —NH—, —O—, —S— or a phosphate; 
 A 1  and A 2  are independently selected from a C 6-30  alkyl, C 6-30  alkenyl, and C 6-30  alkynyl, wherein A 1  and A 2  may be the same or different, 
 or wherein A 1  and A 2  together with the carbon atom to which they are attached form an optionally substituted steroid. 
 
     
     
         5 . The composition of  claim 1  comprising one or more of the compounds selected from:
 a) the compound of formula (I); and/or 
 b) the compound of formulat (XI). 
 
     
     
         6 . The composition of  claim 1 , wherein the biologically active agent is for delivery to the liver or a liver cell and the cationic lipid has a pKa selected from at least:
 (a) about 5.1 to about 7.4;   (b) about 5.3 to about 7.3;   (c) about 5.9 to about 7.0;   (d) about 6.2 to about 6.8; and   (e) about 6.1 or above   
     
     
         7 . The composition of  claim 1 , wherein the biologically active agent is for delivery to a tumor or tumor cells and the cationic lipid has a pKa selected from at least:
 (a) about 5.0 to about 6.7;   (b) about 5.2 to about 6.3;   (c) about 5.4 to about 6.2;   (d) about 5.8 to about 6.1; and   (e) about 6.1 or below.   
     
     
         8 . The composition of  claim 1  further comprising at least one neutral lipid. 
     
     
         9 . The composition of  claim 1 , wherein the composition is optimized by selection of at least one of a stealth lipid, formulation method, N/P ratio, particle size, and molar ratio of the cationic lipid, an optional neutral lipid, helper lipid, stealth lipid and an optional alkyl resorcinol based lipid. 
     
     
         10 . The composition of  claim 5 , wherein the cationic lipid is selected from E0001-E0171 and E0175-E0180. 
     
     
         11 . The composition of  claim 5 , wherein the stealth lipid is selected from S001-S009 and S012-S026. 
     
     
         12 . The composition of  claim 1 , wherein the composition is optimized for at least one parameter including but not limited to individual selection of the pKa of the cationic lipid optimized for the type of cell or organ being targeted, the cationic lipid used, the stealth lipid used, the helper lipid, the neutral lipid used, whether the neutral lipid is present or absent, the ratio of the selected helper lipid, optional neutral lipid, stealth lipid and cationic lipid, the N/P ratio, the particle size, the dosage regiment, the dose given, the formulation method, and the like. 
     
     
         13 . The composition of  claim 1 , further comprising a biologically active agent. 
     
     
         14 . The composition of  claim 13 , wherein the composition comprises a biologically active agent in an amount effective for therapeutic treatment of a disease or disorder. 
     
     
         15 . The composition of  claim 13 , wherein said biologically active agent is selected from the group consisting of antibodies, cholesterol, hormones, antivirals, peptides, polypeptides, proteins, nucleoproteins, chemotherapeutics, low molecular weight drugs, vitamins, co-factors, nucleosides, nucleoside derivatives, nucleotides, oligonucleotides, enzymatic nucleic acids, antisense nucleic acids, triplex forming oligonucleotides, 2,5-A antisense chimeras, allozymes, aptamers, decoy RNA molecules and analogs thereof, and small nucleic acid molecules, such as an RNA interfering agent (RNAi), short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA). 
     
     
         16 . The composition of  claim 15  wherein said biologically active agent is selected from one or more of a nucleoside or nucleoside derivative, an RNAi, an siNA, an RNAi inhibitor, a miRNA, an siRNA and a shRNA. 
     
     
         17 . The composition of  claim 1  that further comprises a pharmaceutically acceptable carrier. 
     
     
         18 . A kit comprising any one or more components of the composition of  claim 1 , and instructions for use. 
     
     
         19 . A method for the treatment of a disease or condition comprising the step of administering a therapeutically effective amount of a composition of  claim 1  to a patient in need thereof. 
     
     
         20 . The method of  claim 19  wherein the disease or condition is a cancer, a disease of the liver, or a disease that is responsive to treatment with an RNAi construct. 
     
     
         21 . A method for the treatment of a disease or condition in a subject in need thereof, the method comprising the step of administering a therapeutically effective amount of a biologically active agent in a formulation comprising one or more compositions of  claim 1 . 
     
     
         22 . The method of  claim 21  wherein the disease or condition is a tumor, a disease of the liver, or a disease that is responsive to treatment with an RNAi construct.