Asialoglycoprotein receptor-mediated delivery of therapeutically active complexes - Patent Application 20070229633

Abstract

ASGP-R binding molecular complexes are provided. The complexes are useful for delivering therapeutically effective amounts of biologically active molecules to target cells and tissues of a subject. Compositions containing the molecular complexes are also provided.

Description

[Technical Field]

[0001] The present disclosure relates to a complex between a therapeutically active molecule and an ASGP-R (asialoglycoprotein receptor) binding ligand that facilitates hepatocyte-selective delivery of the therapeutic molecule. More specifically, the present disclosure relates to a complex between a therapeutically active molecule and an ASGP-R binding ligand that facilitates hepatocyte-selective delivery of the therapeutic molecule in combination with a lipid, as disclosed herein. [Background technology]

[0002] The delivery of therapeutic agents to cells or tissues of human subjects is critical to their therapeutic efficacy and is usually hindered by the compound's limited ability to reach target cells and tissues. Many macromolecules and molecules with a net ionic charge face numerous obstacles to entering cells, and this problem becomes even more complicated when these molecular types need to be delivered to target cell types. Unlike small molecules, macromolecules and molecules with a net ionic charge do not undergo passive diffusion through cell membranes.

[0003] ASGP-R is highly and selectively expressed on the surface of hepatocytes. This receptor was identified and characterized based on its ability to bind to P-linked galactose or GalNAc (N-acetylgalactosamine) residues on proteins. ASGP-R can transport macromolecules across the cell plasma membrane by endocytosis. This receptor has three geometrically arranged carbohydrate-binding domains on its surface. Many molecular constructs consisting of multiple GalNAc molecules can bind to these domains, with each GalNAc unit contributing to the overall binding affinity. Typically, constructs with three GalNAc units spaced approximately 21 Å apart in a triangular arrangement have sufficient binding affinity to affect efficient internalization of conjugated therapeutic molecules.

[0004] While several constructs for attaching therapeutic agents to GalNAc constructs have been developed, the role of the linking moiety and the core structure of the construct is still under investigation. Specifically, the role of several components of these constructs needs to be understood, including stereochemical considerations, linking groups, degradability of the construct, and the flexibility or rigidity of the core to which the linker is attached. Thus, there remains an unmet need for ASGP-R-binding ligand constructs coupled to therapeutic molecules for improved delivery to specific cell types. Summary of the Invention

[0005] The present disclosure relates to a complex of a therapeutically active molecule and an ASGP-R binding ligand that facilitates hepatocyte-selective delivery of the therapeutic molecule. The ASGP-R ligand consists of a modified triantennary display of N-acetylgalactosamine sugar units via a series of amide bonds, using a conformationally restricted spacer unit as an anchor point for both the ligand and the therapeutic molecule. The molecular agent is suitable for optimized presentation of the ligand to target cells. Pharmaceutical compositions containing the ASGP-R binding molecule complex are useful for delivering a therapeutically effective amount of a bioactive molecule to hepatocytes in a patient.

[0006] In one embodiment, disclosed herein is a compound of formula IA: [ka] or a pharmaceutically acceptable salt or solvate thereof; X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n wherein n is 1 to 36, m is 1 to 30, and R N is H, methyl, or CH2F, CHF2, or CF3; Y 1 , Y 2 and Y3 are each independently selected from the group consisting of -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, and P(Z)(OH)O2, where Z is O or S; L 1 , L 2 and L 3 are each independently C1-C 10 Alkyl, -(CH2) e -O-(CH2) f -, -(CH2) e -S-(CH2) f -, -(CH2) e -S(O)2-(CH2) f -, -(CH2) e -NR N -(CH2) f - and -(CH2-CH2-O) k (CH2)2-, wherein e is 1 to 10, f is 1 to 16; k is 1 to 20, and R N is H, methyl, or CH2F, CHF2, or CF3; G 1 , G 2 and G 3 are each independently selected from the group consisting of monosaccharides, monosaccharide derivatives, vitamins, polyols, polysialic acids, and polysialic acid derivatives; 4 is (a)-(CH2) g -O-(CH2) h -or-(CH2) g -NR N -(CH2) h wherein g is 1 to 30, h is 1 to 36, and R N is H, methyl, or CHF, CHF, or CF), (b) amino acids, and (c) -NHC(O)R 2 (In the formula, R 2 is C1-C 10 Alkyl, carbocycle, heterocyclyl, heteroaryl, C1-C 10 Alkyl-Carbocyclic, C1-C 10 Alkyl-heterocyclyl or C1-C 10 alkyl-heteroaryl, wherein R 2is selected from the group consisting of: absent, alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is 1 to 30; j is 1 to 36; and R 3 is hydrogen or alkyl; L 4 -C(O)O-, -C(O)NH-, phosphate, C1-C 10 Alkyl-phosphate, C3-C 10 Alkenyl-phosphate, phosphorothioate, C1-C 10 Alkyl-phosphorothioates, C3-C 10 Alkenyl-phosphorothioates, Boranophosphates, C1-C 10 Alkyl-boranophosphate, C3-C 10 Alkenyl-boranophosphate, -C(O)NH-C3-C 10 Alkyl-phosphate, -C(O)NH-C3-C 10 Alkenyl phosphate, -C(O)O-C1-C 10 Alkyl-phosphate, -C(O)O-C3-C 10 Alkenyl-phosphate, -C(O)NH-C1-C 10 Alkyl-phosphorothioate, -C(O)NH-C3-C10 Alkenyl-phosphorothioate, -C(O)O-C1-C 10 Alkyl-phosphorothioate, -C(O)O-C3-C 10 Alkenyl-phosphorothioate, -C(O)-NH-C-C 10 Alkyl-boranophosphate, -C(O)-NH-C3-C 10 Alkenyl-boranophosphate, -C(O)O-C1-C 10 Alkyl-boranophosphate or -C(O)O-C3-C 10 alkenyl-boranophosphate; and R 1 is a bioactive molecule.

[0007] Additional features and advantages of the subject technology will be set forth in the description that follows, and in part will be obvious from the description, or may be learned by the practice of the subject technology. The advantages of the subject technology will be realized and attained by the structures particularly pointed out in the written description and embodiments herein.

[0008] Various features of exemplary embodiments of the present disclosure are described below with reference to the drawings. The illustrated embodiments are intended to illustrate, but not limit, the present disclosure. The drawings include the following figures: [Brief explanation of the drawings]

[0009] [Figure 1] FVII knockdown data for GalNAc constructs Conjugate 1 (F7 ASO-L-GalNAc) and Conjugate 27 (F7 ASO-L2-GalNAc) are shown. [Figure 2] Figure 1 shows the duration of FVII knockdown for FVII siRNA conjugated to complex 27 (GNAc-2) compared to naked and lipid nanoparticle-encapsulated formulations. DETAILED DESCRIPTION OF THE INVENTION

[0010] Biologically active proteins, e.g., immunoglobulins, and therapeutic agents of the polynucleotide class, e.g., genomic DNA, cDNA, mRNA, and siRNA, antisense oligonucleotides, and even certain small peptides, peptide hormones, and antibiotics, represent classes of molecules where targeted delivery to tissues of interest by diffusion across cell membranes faces significant obstacles.

[0011] Receptor-mediated endocytosis (RME) is a well-known biological mechanism by which cells internalize extracellular molecules. This process requires the binding of a given cell surface receptor to its cognate ligand, which may be expressed as an epitope on the surface of the molecule to be internalized. This cognate ligand can be used as a targeting ligand to deliver therapeutically relevant molecules to specific target cells. Therefore, receptor-ligand combinations can be utilized with therapeutically active molecules by conjugating the targeting ligand to the active molecule, achieving significantly improved targeted delivery to specific cell types in a subject. Some prominent examples of receptor-mediated endocytosis systems are those that recognize sugars such as galactose, mannose, and mannose-6-phosphate, or peptides and proteins such as transferrin and asialoglycoprotein.

[0012] ASGP-R is highly and selectively expressed on the surface of liver cells called hepatocytes. It was identified and characterized based on its ability to bind to P-linked galactose or GalNAc residues on proteins. In this manner, ASGP-R can transport macromolecules and charged molecules across the cell plasma membrane by endocytosis.

[0013] Disclosed herein are compounds and pharmaceutical compositions comprising ASGP-R binding molecule complexes that are useful for delivering therapeutically effective amounts of bioactive molecules to liver cells in a subject.

[0014] In one embodiment, disclosed herein is a compound of formula IA: [ka] or a pharmaceutically acceptable salt or solvate thereof, wherein X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n wherein n is 1 to 36, m is 1 to 30, and R N is H, methyl, or CH2F, CHF2, or CF3; Y 1 , Y 2 and Y 3 are each independently selected from the group consisting of -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, and P(Z)(OH)O2, where Z is O or S; L 1 , L 2 and L 3 are each independently C1-C 10 Alkyl, -(CH2) e -O-(CH2) f -, -(CH2) e -S-(CH2) f -, -(CH2) e -S(O)2-(CH2) f -, -(CH2) e -NR N -(CH2) f - and -(CH2-CH2-O) k (CH2)2-, wherein e is 1 to 10, f is 1 to 16, k is 1 to 20, and R N is H, methyl, or CH2F, CHF2, or CF3; G 1 , G 2 and G 3 are each independently selected from the group consisting of monosaccharides, monosaccharide derivatives, vitamins, polyols, polysialic acids, and polysialic acid derivatives; 4 is (a)-(CH2) g-O-(CH2) h -or-(CH2) g -NR N -(CH2) h wherein g is 1 to 30, h is 1 to 36, and R N is H, methyl, or CHF, CHF, or CF), (b) amino acids, and (c) -NHC(O)R 2 (In the formula, R 2 is C1-C 10 Alkyl, carbocycle, heterocyclyl, heteroaryl, C1-C 10 Alkyl-Carbocyclic, C1-C 10 Alkyl-heterocyclyl or C1-C 10 alkyl-heteroaryl, and wherein R 2 is optionally substituted), and Q is absent, alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is 1 to 30; j is 1 to 36; and R 3 is hydrogen or alkyl; L 4 -C(O)O-, -C(O)NH-, phosphate, C1-C 10 Alkyl-phosphate, C3-C 10Alkenyl-phosphate, phosphorothioate, C1-C 10 Alkyl-phosphorothioates, C3-C 10 Alkenyl-phosphorothioates, Boranophosphates, C1-C 10 Alkyl-boranophosphate, C3-C 10 Alkenyl-boranophosphate, -C(O)NH-C-C 10 Alkyl-phosphate, -C(O)NH-C3-C 10 Alkenyl phosphate, -C(O)O-C1-C 10 Alkyl-phosphate, -C(O)O-C3-C 10 Alkenyl-phosphate, -C(O)NH-C1-C 10 Alkyl-phosphorothioate, -C(O)NH-C3-C 10 Alkenyl-phosphorothioate, -C(O)O-C1-C 10 Alkyl-phosphorothioate, -C(O)O-C3-C 10 Alkenyl-phosphorothioate, -C(O)-NH-C-C 10 Alkyl-boranophosphate, -C(O)-NH-C3-C 10 Alkenyl-boranophosphate, -C(O)O-C1-C 10 Alkyl-boranophosphate or -C(O)O-C3-C 10 alkenyl-boranophosphate; and R 1 is a biologically active molecule; and a lipid of formula II, III, or IV, as disclosed herein.

[0015] In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n-, wherein m is 1 to 36, 1 to 35, 1 to 34, 1 to 33, 1 to 32, 1 to 31, 1 to 30, 1 to 29, 1 to 28, 1 to 27, 1 to 26, 1 to 25, 1 to 24, 1 to 23, 1 to 22, 1 to 21, 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2 or 1, and R N is H, methyl, or CHF, CHF, or CF. In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n -, wherein m is 1 to 30, 1 to 29, 1 to 28, 1 to 27, 1 to 26, 1 to 25, 1 to 24, 1 to 23, 1 to 22, 1 to 21, 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, and 1. In some embodiments, X 1 , X 2 and X 3 are each independently (-CH2) m -O-CH2-, where m is 1 to 4. In some embodiments, X 1 , X 2 and X 3 are each independently (—CH)—O—CH—. In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl or -CH2-.

[0016] In some embodiments, Y 1 , Y 2 and Y 3 are each —NHC(O)— or —C(O)NH—. In some embodiments, Y 1 , Y 2 and Y 3 are each -NHC(O)-.

[0017] In some embodiments, L 1 , L 2 and L 3 are C1-C 10 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. In some embodiments, L 1 , L 2 and L 3 are each independently C3-C8 alkyl or -(CH2-CH2-O) k (CH2)2-, where k is 1 to 10. In some embodiments, L 1 , L 2 and L 3 are each independently -(CH2-CH2-O) k (CH2)2-, where k is 2 to 4. In some embodiments, L 1 , L 2 and L 3 are each independently -(CH2-CH2-O) k (CH2)2-, where k is 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1. In some embodiments, L 1 , L 2 and L 3 are each -(CH2-CH2-O)(CH2)2-.

[0018] In some embodiments, G 1 , G 2 and G 3are each independently folic acid, ribose, retinol, niacin, riboflavin, biotin, glucose, mannose, fucose, sucrose, lactose, mannose-6-phosphate, N-acetylgalactosamine, N-acetylglucosamine, sialic acid, sialic acid derivatives, allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, fucitol, fucosamine, fucose, fuculose, galactosamine, galactosaminitol, galactose, glucosamine, glucosaminitol, glucose-6 In some embodiments, the sugar is selected from the group consisting of phosphate, guloseglyceraldehyde, glycero-mannoseptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, psicose, quinovose, quinovosamine, rhamnitol, rhamnosamine, rhamnose, ribulose, sedoheptulose, sorbose, tagatose, talose, threose, xylose, and xylulose. 1 , G 2 and G 3 are each N-acetylgalactosamine.

[0019] In some embodiments, X 4 teeth, [ka] wherein X 4 is optionally substituted.

[0020] In some embodiments, X 4 is -NHC(O)R 2 where R 2 is a carbocycle, heterocyclyl, or heteroaryl, where R 2 is optionally substituted. In some embodiments, X 4 is -NHC(O)R 2 where R 2 is a carbocycle, heterocyclyl, or heteroaryl, where R 2 is optionally substituted with alkyl, alkoxy, or amine. In some embodiments, X4 teeth, [ka] is.

[0021] In some embodiments, Q is alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is 1 to 10 and j is 1 to 10; and R 3 is hydrogen or alkyl. In some embodiments, Q is alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i-C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is 1 to 10 and j is 1 to 10; i is 1 to 9 and j is 1 to 9; i is 1 to 8 and j is 1 to 8; i is 1 to 7 and j is 1 to 7; i is 1 to 6 and j is 1 to 6; i is 1 to 5 and j is 1 to 5; i is 1 to 5 and j is 1 to 4; i is 1 to 3 and j is 1 to 3; i is 1 to 2 and j is 1 to 2; or i is 1 and j is 1.

[0022] In some embodiments, Q is -C(O)-(CH) 1-10 - and L 4 is -C(O)NH-(CH2) 1-10 In some embodiments, Q is -C(O)-(CH) 1-9 - and L 4 is -C(O)NH-(CH2) 1-9 -phosphate; Q is -C(O)-(CH) 1-8 - and L 4 is -C(O)NH-(CH2) 1-8 -phosphate; Q is -C(O)-(CH) 1-7 - and L 4 is -C(O)NH-(CH2) 1-7 -phosphate; Q is -C(O)-(CH) 1-6 - and L 4 is -C(O)NH-(CH2) 1-6 -phosphate; Q is -C(O)-(CH) 1-5 - and L 4 is -C(O)NH-(CH2) 1-5 -phosphate; Q is -C(O)-(CH) 1-4 - and L 4 is -C(O)NH-(CH2) 1-4-phosphate; Q is -C(O)-(CH) 1-3 - and L 4 is -C(O)NH-(CH2) 1-3 -phosphate; Q is -C(O)-(CH) 1-2 - and L 4 is -C(O)NH-(CH2) 1-2 -phosphate; or Q is -C(O)-(CH)-, and L 4 is —C(O)NH—(CH)-phosphate. In some embodiments, L 4 is -C(O)NH-(CH2) 1-10 In some embodiments, L 4 is -C(O)NH-(CH2) 1-9 -phosphate, -C(O)NH-(CH2) 1-8 -phosphate, -C(O)NH-(CH2) 1-7 -phosphate, -C(O)NH-(CH2) 1-6 -phosphate, -C(O)NH-(CH2) 1-5 -phosphate, -C(O)NH-(CH2) 1-4 -phosphate, -C(O)NH-(CH2) 1-3 -phosphate, -C(O)NH-(CH2) 1-2 In some embodiments, Q is —C(O)—(CH)—, and L 4 is —C(O)NH—(CH2)6-phosphate.

[0023] In some embodiments, L 4 -C(O)O-, -C(O)NH-, phosphate, C1-C 10 Alkyl-phosphate, C3-C 10 Alkenyl-phosphate, phosphorothioate, C1-C 10 Alkyl-phosphorothioates, C3-C 10 Alkenyl-phosphorothioates, phosphorothioates, C1-C 10 Alkyl-phosphorothioates, C3-C 10Alkenyl-phosphorothioates, Boranophosphates, C1-C 10 Alkyl-boranophosphate, C3-C 10 Alkenyl-boranophosphate, -C(O)NH-C-C 10 Alkyl-phosphate, -C(O)O-C1-C 10 Alkyl-phosphate, phosphorothioate, -C(O)NH-C1-C 10 Alkyl-phosphorothioate, -C(O)O-C1-C 10 Alkyl-phosphorothioate, boranophosphate, -C(O)-NH-C1-C 10 Alkyl-boranophosphate or -C(O)O-C1-C 10 Alkyl-boranophosphate, -C(O)NH-C3-C 10 Alkenyl-phosphate, -C(O)O-C3-C 10 Alkenyl-phosphate, -C(O)NH-C3-C 10 Alkenyl-phosphorothioate, -C(O)O-C3-C 10 Alkenyl-phosphorothioate, -C(O)-NH-C3-C 10 Alkenyl-boranophosphate or -C(O)O-C3-C 10 It is an alkenyl-boranophosphate.

[0024] In some embodiments, L 4 is C1-C 20 Alkyl-phosphates, C1-C 19 Alkyl-phosphates, C1-C 18 Alkyl-phosphates, C1-C 17 Alkyl-phosphates, C1-C 16 Alkyl-phosphates, C1-C 15 Alkyl-phosphates, C1-C 14 Alkyl-phosphates, C1-C 13 Alkyl-phosphates, C1-C 12 Alkyl-phosphates, C1-C 11 Alkyl-phosphates, C1-C 10alkyl-phosphate, C1-C9 alkyl-phosphate, C1-C8 alkyl-phosphate, C1-C7 alkyl-phosphate, C1-C6 alkyl-phosphate, C1-C5 alkyl-phosphate, C1-C4 alkyl-phosphate, C1-C3 alkyl-phosphate, C1-C2 alkyl-phosphate, or -CH2-phosphate. 4 is C1-C 20 Alkyl-phosphorothioates, C1-C 19 Alkyl-phosphorothioates, C1-C 18 Alkyl-phosphorothioates, C1-C 17 Alkyl-phosphorothioates, C1-C 16 Alkyl-phosphorothioates, C1-C 15 Alkyl-phosphorothioates, C1-C 14 Alkyl-phosphorothioates, C1-C 13 Alkyl-phosphorothioates, C1-C 12 Alkyl-phosphorothioates, C1-C 11 Alkyl-phosphorothioates, C1-C 10 alkyl-phosphorothioate, C1-C9 alkyl-phosphorothioate, C1-C8 alkyl-phosphorothioate, C1-C7 alkyl-phosphorothioate, C1-C6 alkyl-phosphorothioate, C1-C5 alkyl-phosphorothioate, C1-C4 alkyl-phosphorothioate, C1-C3 alkyl-phosphorothioate, C1-C2 alkyl-phosphorothioate or -CH2-phosphorothioate. 4 is C1-C 20 Alkyl-boranophosphates, C1-C 19 Alkyl-boranophosphates, C1-C 18 Alkyl-boranophosphates, C1-C 17 Alkyl-boranophosphates, C1-C 16 Alkyl-boranophosphates, C1-C 15 Alkyl-boranophosphates, C1-C 14 Alkyl-boranophosphates, C1-C 13Alkyl-boranophosphates, C1-C 12 Alkyl-boranophosphates, C1-C 11 Alkyl-boranophosphates, C1-C 10 alkyl-boranophosphate, C1-C9 alkyl-boranophosphate, C1-C8 alkyl-boranophosphate, C1-C7 alkyl-boranophosphate, C1-C6 alkyl-boranophosphate, C1-C5 alkyl-boranophosphate, C1-C4 alkyl-boranophosphate, C1-C3 alkyl-boranophosphate, C1-C2 alkyl-boranophosphate or -CH2-boranophosphate.

[0025] In some embodiments, L 4 is C3-C 20 Alkenyl Phosphates, C3-C 19 Alkenyl Phosphates, C3-C 18 Alkenyl Phosphates, C3-C 17 Alkenyl Phosphates, C3-C 16 Alkenyl Phosphates, C3-C 15 Alkenyl Phosphates, C3-C 14 Alkenyl Phosphates, C3-C 13 Alkenyl Phosphates, C3-C 12 Alkenyl Phosphates, C3-C 11 Alkenyl Phosphates, C3-C 10 alkenyl-phosphate, C3-C9 alkenyl-phosphate, C3-C8 alkenyl-phosphate, C3-C7 alkenyl-phosphate, C3-C6 alkenyl-phosphate, C3-C5 alkenyl-phosphate, C3-C4 alkenyl-phosphate, or C3 alkenyl-phosphate. 4 is C3-C 20 Alkenyl-phosphorothioates, C3-C 19 Alkenyl-phosphorothioates, C3-C 18 Alkenyl-phosphorothioates, C3-C 17 Alkenyl-phosphorothioates, C3-C 16Alkenyl-phosphorothioates, C3-C 15 Alkenyl-phosphorothioates, C3-C 14 Alkenyl-phosphorothioates, C3-C 13 Alkenyl-phosphorothioates, C3-C 12 Alkenyl-phosphorothioates, C3-C 11 Alkenyl-phosphorothioates, C3-C 10 In some embodiments, L is an alkenyl-phosphorothioate, a C3-C9 alkenyl-phosphorothioate, a C3-C8 alkenyl-phosphorothioate, a C3-C7 alkenyl-phosphorothioate, a C3-C6 alkenyl-phosphorothioate, a C3-C5 alkenyl-phosphorothioate, a C3-C4 alkenyl-phosphorothioate, or a C3 alkenyl-phosphorothioate. 4 is C3-C 20 Alkenyl-boranophosphate, C3-C 19 Alkenyl-boranophosphate, C3-C 18 Alkenyl-boranophosphate, C3-C 17 Alkenyl-boranophosphate, C3-C 16 Alkenyl-boranophosphate, C3-C 15 Alkenyl-boranophosphate, C3-C 14 Alkenyl-boranophosphate, C3-C 13 Alkenyl-boranophosphate, C3-C 12 Alkenyl-boranophosphate, C3-C 11 Alkenyl-boranophosphate, C3-C 10 alkenyl-boranophosphate, C3-C9 alkenyl-boranophosphate, C3-C8 alkenyl-boranophosphate, C3-C7 alkenyl-boranophosphate, C3-C6 alkenyl-boranophosphate, C3-C5 alkenyl-boranophosphate, C3-C4 alkenyl-boranophosphate, or C3 alkenyl-boranophosphate.

[0026] In some embodiments, L 4 is -C(O)NH-C1-C 10alkyl-phosphate, —C(O)NH—C1-C9 alkyl-phosphate, —C(O)NH—C1-C8 alkyl-phosphate, —C(O)NH—C1-C7 alkyl-phosphate, —C(O)NH—C1-C6 alkyl-phosphate, —C(O)NH—C1-C5 alkyl-phosphate, —C(O)NH—C1-C4 alkyl-phosphate, —C(O)NH—C1-C3 alkyl-phosphate, —C(O)NH—C1-C2 alkyl-phosphate, or —C(O)NH—CH2-phosphate.

[0027] In some embodiments, L 4 is -C(O)NH-C3-C 10 alkenyl-phosphate, -C(O)NH-C3-C9 alkenyl-phosphate, -C(O)NH-C3-C8 alkenyl-phosphate, -C(O)NH-C3-C7 alkenyl-phosphate, -C(O)NH-C3-C6 alkenyl-phosphate, -C(O)NH-C3-C5 alkenyl-phosphate, -C(O)NH-C3-C4 alkenyl-phosphate, or -C(O)NH-C3 alkenyl-phosphate.

[0028] In some embodiments, L 4 is -C(O)O-C1-C 10 alkyl-phosphate, —C(O)O—C1-C9 alkyl-phosphate, —C(O)O—C1-C8 alkyl-phosphate, —C(O)O—C1-C7 alkyl-phosphate, —C(O)O—C1-C6 alkyl-phosphate, —C(O)O—C1-C5 alkyl-phosphate, —C(O)O—C1-C4 alkyl-phosphate, —C(O)O—C1-C3 alkyl-phosphate, —C(O)O—C1-C2 alkyl-phosphate, or —C(O)O—CH2-phosphate.

[0029] In some embodiments, L 4 is -C(O)O-C3-C 10alkenyl-phosphate, -C(O)O-C3-C9 alkenyl-phosphate, -C(O)O-C3-C8 alkenyl-phosphate, -C(O)O-C3-C7 alkenyl-phosphate, -C(O)O-C3-C6 alkenyl-phosphate, -C(O)O-C3-C5 alkenyl-phosphate, -C(O)O-C3-C4 alkenyl-phosphate, or -C(O)O-C3 alkenyl-phosphate.

[0030] In some embodiments, L 4 is -C(O)NH-C1-C 10 alkyl-phosphorothioate, -C(O)NH-C1-C9 alkyl-phosphorothioate, -C(O)NH-C1-C8 alkyl-phosphorothioate, -C(O)NH-C1-C7 alkyl-phosphorothioate, -C(O)NH-C1-C6 alkyl-phosphorothioate, -C(O)NH-C1-C5 alkyl-phosphorothioate, -C(O)NH-C1-C4 alkyl-phosphorothioate, -C(O)NH-C1-C3 alkyl-phosphorothioate, -C(O)NH-C1-C2 alkyl-phosphorothioate, or -C(O)NH-CH2-phosphorothioate.

[0031] In some embodiments, L 4 is -C(O)NH-C3-C 10 alkenyl-phosphorothioate, -C(O)NH-C3-C9 alkenyl-phosphorothioate, -C(O)NH-C3-C8 alkenyl-phosphorothioate, -C(O)NH-C3-C7 alkenyl-phosphorothioate, -C(O)NH-C3-C6 alkenyl-phosphorothioate, -C(O)NH-C3-C5 alkenyl-phosphorothioate, -C(O)NH-C3-C4 alkenyl-phosphorothioate, or -C(O)NH-C3 alkenyl-phosphorothioate.

[0032] In some embodiments, L 4 is -C(O)O-C1-C 10alkyl-phosphorothioate, -C(O)O-C1-C9 alkyl-phosphorothioate, -C(O)O-C1-C8 alkyl-phosphorothioate, -C(O)O-C1-C7 alkyl-phosphorothioate, -C(O)O-C1-C6 alkyl-phosphorothioate, -C(O)O-C1-C5 alkyl-phosphorothioate, -C(O)O-C1-C4 alkyl-phosphorothioate, -C(O)O-C1-C3 alkyl-phosphorothioate, -C(O)O-C1-C2 alkyl-phosphorothioate, or -C(O)O-CH2-phosphorothioate.

[0033] In some embodiments, L 4 is -C(O)O-C3-C 10 alkenyl-phosphorothioate, -C(O)O-C3-C9 alkenyl-phosphorothioate, -C(O)O-C3-C8 alkenyl-phosphorothioate, -C(O)O-C3-C7 alkenyl-phosphorothioate, -C(O)O-C3-C6 alkenyl-phosphorothioate, -C(O)O-C3-C5 alkenyl-phosphorothioate, -C(O)O-C3-C4 alkenyl-phosphorothioate, or -C(O)O-C3 alkenyl-phosphorothioate.

[0034] In some embodiments, L 4 is -C(O)-NH-C1-C 10 alkyl-boranophosphate, —C(O)—NH—C1-C9 alkyl-boranophosphate, —C(O)—NH—C1-C8 alkyl-boranophosphate, —C(O)—NH—C1-C7 alkyl-boranophosphate, —C(O)—NH—C1-C6 alkyl-boranophosphate, —C(O)—NH—C1-C5 alkyl-boranophosphate, —C(O)—NH—C1-C4 alkyl-boranophosphate, —C(O)—NH—C1-C3 alkyl-boranophosphate, —C(O)—NH—C1-C2 alkyl-boranophosphate, or —C(O)—NH—CH2-boranophosphate.

[0035] In some embodiments, L 4is -C(O)-NH-C3-C 10 alkenyl-boranophosphate, -C(O)-NH-C3-C9 alkenyl-boranophosphate, -C(O)-NH-C3-C8 alkenyl-boranophosphate, -C(O)-NH-C3-C7 alkenyl-boranophosphate, -C(O)-NH-C3-C6 alkenyl-boranophosphate, -C(O)-NH-C3-C5 alkenyl-boranophosphate, -C(O)-NH-C3-C4 alkenyl-boranophosphate, or -C(O)-NH-C3 alkenyl-boranophosphate.

[0036] In some embodiments, L 4 is -C(O)O-C1-C 10 alkyl-boranophosphate, —C(O)O—C1-C9 alkyl-boranophosphate, —C(O)O—C1-C8 alkyl-boranophosphate, —C(O)O—C1-C7 alkyl-boranophosphate, —C(O)O—C1-C6 alkyl-boranophosphate, —C(O)O—C1-C5 alkyl-boranophosphate, —C(O)O—C1-C4 alkyl-boranophosphate, —C(O)O—C1-C3 alkyl-boranophosphate, —C(O)O—C1-C2 alkyl-boranophosphate, or —C(O)O—CH2-boranophosphate.

[0037] In some embodiments, L 4 is -C(O)O-C3-C 10 alkenyl-boranophosphate, —C(O)O—C3-C9 alkenyl-boranophosphate, —C(O)O—C3-C8 alkenyl-boranophosphate, —C(O)O—C3-C7 alkenyl-boranophosphate, —C(O)O—C3-C6 alkenyl-boranophosphate, —C(O)O—C3-C5 alkenyl-boranophosphate, —C(O)O—C3-C4 alkenyl-boranophosphate, or —C(O)O—C3 alkenyl-boranophosphate.

[0038] In some embodiments, R 1is selected from the group consisting of pentafluorophenyl, tetrafluorophenyl, succinimide, maleimide, azide, pyridyldithiol, dimethyl methylphosphonate, chiral dimethyl methylphosphonate, helper lipids, and nucleic acids. 1 is an ASO (antisense oligonucleotide), siRNA (small interfering RNA), miRNA (microRNA), microRNA mimic, AMO (anti-miRNA oligonucleotide), long non-coding RNA, PNA (peptide nucleic acid), helper lipid, or PMO (phosphorodiamidate morpholino oligomer), wherein the nucleic acid is optionally modified. In some embodiments, R 1 is an ASO (antisense oligonucleotide). In some embodiments, R 1 is a PEG-lipid conjugate. In some embodiments, the PEG-lipid conjugate is PEG-DMG. In some embodiments, the PEG-DMG is PEG2000-DMG.

[0039] In some embodiments, disclosed herein is a compound having the formula: [ka] wherein R 1 is an ASO (antisense oligonucleotide), siRNA (small interfering RNA), miRNA (microRNA), microRNA mimic, AMO (anti-miRNA oligonucleotide), long non-coding RNA, PNA (peptide nucleic acid), helper lipid, or PMO (phosphorodiamidate morpholino oligomer), wherein the nucleic acid is optionally modified.

[0040] In some embodiments, the compound of formula IA is [ka] [ka] [ka] [ka] [ka] [ka] [ka] wherein: [ka] is an oligonucleotide.

[0041] In another embodiment, disclosed herein is a compound of formula IB [ka] or a pharmaceutically acceptable salt or solvate thereof, wherein X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n wherein n is 1 to 36, m is 1 to 30, and R N is H, methyl, or CH2F, CHF2, or CF3; Y 1 , Y 2 and Y 3 are each independently selected from the group consisting of -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, and P(Z)(OH)O2, where Z is O or S; L 1 , L 2 and L 3 are each independently C1-C 10 Alkyl, -(CH2) e-O-(CH2) f -, -(CH2) e -S-(CH2) f -, -(CH2) e -S(O)2-(CH2) f -, -(CH2) e -NR N -(CH2) f - and -(CH2-CH2-O) k (CH2)2-, wherein e is 1 to 10; f is 1 to 16, k is 1 to 20, and R N is H, methyl, or CH2F, CHF2, or CF3; G 1 , G 2 and G 3 are each independently selected from the group consisting of monosaccharides, monosaccharide derivatives, vitamins, polyols, polysialic acids, and polysialic acid derivatives; 4 is (a)-(CH2) g -O-(CH2) h -or-(CH2) g -NR N -(CH2) h wherein g is 1 to 30, h is 1 to 36, and R N is H, methyl, or CHF, CHF, or CF), (b) amino acids, and (c) -NHC(O)R 2 (In the formula, R 2 is C1-C 10 Alkyl, carbocycle, heterocyclyl, heteroaryl, C1-C 10 Alkyl-Carbocyclic, C1-C 10 Alkyl-heterocyclyl or C1-C 10 alkyl-heteroaryl, and wherein R 2 is optionally substituted), and Q is selected from the group consisting of alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j-, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j -, -(CH2) i -C(O)S-(CH2) j -,or [ka] In the formula, H 1 is carbocycle, heterocyclyl, or heteroaryl; H 1 is optionally substituted; i is 1 to 30 and j is 1 to 36; R 3 is hydrogen or alkyl; W 1 and W 2 are each independently selected from —CH— and O; v is 1 to 6; Y is hydrogen or methyl; and T is C-C 10 Alkyl or C2-C 10 alkenyl; L 4 -C(O)O-, -C(O)NH-, phosphate, C1-C 10 Alkyl-phosphate, C3-C 10 Alkenyl-phosphate, phosphorothioate, C1-C 10 Alkyl-phosphorothioates, C3-C 10 Alkenyl-phosphorothioates, Boranophosphates, C1-C 10 Alkyl-boranophosphate, C3-C 10 Alkenyl-boranophosphate, -C(O)NH-C-C 10 Alkyl-phosphate, -C(O)NH-C3-C 10 Alkenyl phosphate, -C(O)O-C1-C 10 Alkyl-phosphate, -C(O)O-C3-C 10Alkenyl-phosphate, -C(O)NH-C1-C 10 Alkyl-phosphorothioate, -C(O)NH-C3-C 10 Alkenyl-phosphorothioate, -C(O)O-C1-C 10 Alkyl-phosphorothioate, -C(O)O-C3-C 10 Alkenyl-phosphorothioate, -C(O)-NH-C-C 10 Alkyl-boranophosphate, -C(O)-NH-C3-C 10 Alkenyl-boranophosphate, -C(O)O-C1-C 10 Alkyl-boranophosphate or -C(O)O-C3-C 10 alkenyl-boranophosphate; and R 1 is a bioactive molecule.

[0042] In some embodiments, W 1 and W 2 are each independently selected from —CH2— and O, wherein v is 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or —CH2—.

[0043] In some embodiments, T is C1-C 10 In some embodiments, T is C2-C3 alkyl, C1-C4 alkyl, C1-C5 alkyl, C1-C6 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. 10 alkenyl, C2-C9 alkenyl, C2-C8 alkenyl, C2-C7 alkenyl, C2-C6 alkenyl, C2-C5 alkenyl, C2-C4 alkenyl, C2-C3 alkenyl or -CH=CH-.

[0044] In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NRN -(CH2) n -, where n is 1 to 36, 1 to 35, 1 to 34, 1 to 33, 1 to 32, 1 to 31, 1 to 30, 1 to 29, 1 to 28, 1 to 27, 1 to 26, 1 to 25, 1 to 24, 1 to 23, 1 to 22, 1 to 21, 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, and 1. In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n -, wherein m is 1 to 30, 1 to 29, 1 to 28, 1 to 27, 1 to 26, 1 to 25, 1 to 24, 1 to 23, 1 to 22, 1 to 21, 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, and 1. In some embodiments, X 1 , X 2 and X 3 are each independently (-CH2) m -O-CH2-, where m is 1 to 4. In some embodiments, X 1 , X 2 and X 3 are each independently (—CH)—O—CH—. In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl or -CH2-.

[0045] In some embodiments, Y 1, Y 2 and Y 3 are each —NHC(O)— or —C(O)NH—. In some embodiments, Y 1 , Y 2 and Y 3 are each -NHC(O)-.

[0046] In some embodiments, L 1 , L 2 and L 3 are C1-C 10 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. In some embodiments, L 1 , L 2 and L 3 are each independently C3-C8 alkyl or -(CH2-CH2-O) k (CH2)2-, where k is 1 to 10. In some embodiments, L 1 , L 2 and L 3 are each independently -(CH2-CH2-O) k (CH2)2-, where k is 2 to 4. In some embodiments, L 1 , L 2 and L 3 are each independently -(CH2-CH2-O) k (CH2)2-, where k is 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1. In some embodiments, L 1 , L 2 and L 3 are each independently —(CH—CH—O)(CH)—.

[0047] In some embodiments, G 1 , G 2 and G 3are each independently folic acid, ribose, retinol, niacin, riboflavin, biotin, glucose, mannose, fucose, sucrose, lactose, mannose-6-phosphate, N-acetylgalactosamine, N-acetylglucosamine, sialic acid, sialic acid derivatives, allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, fucitol, fucosamine, fucose, fuculose, galactosamine, galactosaminitol, galactose, glucosamine, glucosaminitol, glucose-6 In some embodiments, the sugar is selected from the group consisting of phosphate, guloseglyceraldehyde, glycero-mannoseptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, psicose, quinovose, quinovosamine, rhamnitol, rhamnosamine, rhamnose, ribulose, sedoheptulose, sorbose, tagatose, talose, threose, xylose, and xylulose. 1 , G 2 and G 3 are each N-acetylgalactosamine.

[0048] In some embodiments, X 4 teeth, [ka] wherein X 4 is optionally substituted.

[0049] In some embodiments, X 4 is -NHC(O)R 2 where R 2 is a carbocycle, heterocyclyl, or heteroaryl, where R 2 is optionally substituted. In some embodiments, X 4 is -NHC(O)R 2 where R 2 is a carbocycle, heterocyclyl, or heteroaryl, where R 2 is optionally substituted with alkyl, alkoxy or amine.

[0050] In some embodiments, X 4 teeth, [ka] is.

[0051] In some embodiments, Q is alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is 1 to 10, j is 1 to 10, and wherein R 3 is hydrogen or alkyl. In some embodiments, Q is alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i-NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is 1 to 10 and j is 1 to 10; i is 1 to 9 and j is 1 to 9; i is 1 to 8 and j is 1 to 8; i is 1 to 7 and j is 1 to 7; i is 1 to 6 and j is 1 to 6; i is 1 to 5 and j is 1 to 5; i is 1 to 5 and j is 1 to 4; i is 1 to 3 and j is 1 to 3; i is 1 to 2 and j is 1 to 2; or i is 1 and j is 1.

[0052] In some embodiments, L 4 -C(O)O-, -C(O)NH-, phosphate, C1-C 10 Alkyl-phosphate, C3-C 10 Alkenyl-phosphate, phosphorothioate, C1-C 10 Alkyl-phosphorothioates, C3-C 10 Alkenyl-phosphorothioates, Boranophosphates, C1-C 10 Alkyl-boranophosphate, C3-C 10 Alkenyl-boranophosphate, -C(O)NH-C-C 10 Alkyl-phosphate, -C(O)NH-C3-C 10 Alkenyl phosphate, -C(O)O-C1-C 10 Alkyl-phosphate, -C(O)O-C3-C 10 Alkenyl-phosphate, -C(O)NH-C1-C 10 Alkyl-phosphorothioate, -C(O)NH-C3-C 10 Alkenyl-phosphorothioate, -C(O)O-C1-C 10 Alkyl-phosphorothioate, -C(O)O-C3-C 10 Alkenyl-phosphorothioate, -C(O)-NH-C-C10 Alkyl-boranophosphate, -C(O)-NH-C3-C 10 Alkenyl-boranophosphate, -C(O)O-C1-C 10 Alkyl-boranophosphate or -C(O)O-C3-C 10 It is an alkenyl-boranophosphate.

[0053] In some embodiments, L 4 is C1-C 20 Alkyl-phosphates, C1-C 19 Alkyl-phosphates, C1-C 18 Alkyl-phosphates, C1-C 17 Alkyl-phosphates, C1-C 16 Alkyl-phosphates, C1-C 15 Alkyl-phosphates, C1-C 14 Alkyl-phosphates, C1-C 13 Alkyl-phosphates, C1-C 12 Alkyl-phosphates, C1-C 11 Alkyl-phosphates, C1-C 10 alkyl-phosphate, C1-C9 alkyl-phosphate, C1-C8 alkyl-phosphate, C1-C7 alkyl-phosphate, C1-C6 alkyl-phosphate, C1-C5 alkyl-phosphate, C1-C4 alkyl-phosphate, C1-C3 alkyl-phosphate, C1-C2 alkyl-phosphate, or -CH2-phosphate. 4 is C1-C 20 Alkyl-phosphorothioates, C1-C 19 Alkyl-phosphorothioates, C1-C 18 Alkyl-phosphorothioates, C1-C 17 Alkyl-phosphorothioates, C1-C 16 Alkyl-phosphorothioates, C1-C 15 Alkyl-phosphorothioates, C1-C 14 Alkyl-phosphorothioates, C1-C 13 Alkyl-phosphorothioates, C1-C 12Alkyl-phosphorothioates, C1-C 11 Alkyl-phosphorothioates, C1-C 10 alkyl-phosphorothioate, C1-C9 alkyl-phosphorothioate, C1-C8 alkyl-phosphorothioate, C1-C7 alkyl-phosphorothioate, C1-C6 alkyl-phosphorothioate, C1-C5 alkyl-phosphorothioate, C1-C4 alkyl-phosphorothioate, C1-C3 alkyl-phosphorothioate, C1-C2 alkyl-phosphorothioate or -CH2-phosphorothioate. 4 is C1-C 20 Alkyl-boranophosphates, C1-C 19 Alkyl-boranophosphates, C1-C 18 Alkyl-boranophosphates, C1-C 17 Alkyl-boranophosphates, C1-C 16 Alkyl-boranophosphates, C1-C 15 Alkyl-boranophosphates, C1-C 14 Alkyl-boranophosphates, C1-C 13 Alkyl-boranophosphates, C1-C 12 Alkyl-boranophosphates, C1-C 11 Alkyl-boranophosphates, C1-C 10 alkyl-boranophosphate, C1-C9 alkyl-boranophosphate, C1-C8 alkyl-boranophosphate, C1-C7 alkyl-boranophosphate, C1-C6 alkyl-boranophosphate, C1-C5 alkyl-boranophosphate, C1-C4 alkyl-boranophosphate, C1-C3 alkyl-boranophosphate, C1-C2 alkyl-boranophosphate or -CH2-boranophosphate.

[0054] In some embodiments, L 4 is C3-C 20 Alkenyl Phosphates, C3-C 19 Alkenyl Phosphates, C3-C 18 Alkenyl Phosphates, C3-C 17Alkenyl Phosphates, C3-C 16 Alkenyl Phosphates, C3-C 15 Alkenyl Phosphates, C3-C 14 Alkenyl Phosphates, C3-C 13 Alkenyl Phosphates, C3-C 12 Alkenyl Phosphates, C3-C 11 Alkenyl Phosphates, C3-C 10 alkenyl-phosphate, C3-C9 alkenyl-phosphate, C3-C8 alkenyl-phosphate, C3-C7 alkenyl-phosphate, C3-C6 alkenyl-phosphate, C3-C5 alkenyl-phosphate, C3-C4 alkenyl-phosphate, or C3 alkenyl-phosphate. 4 is C3-C 20 Alkenyl-phosphorothioates, C3-C 19 Alkenyl-phosphorothioates, C3-C 18 Alkenyl-phosphorothioates, C3-C 17 Alkenyl-phosphorothioates, C3-C 16 Alkenyl-phosphorothioates, C3-C 15 Alkenyl-phosphorothioates, C3-C 14 Alkenyl-phosphorothioates, C3-C 13 Alkenyl-phosphorothioates, C3-C 12 Alkenyl-phosphorothioates, C3-C 11 Alkenyl-phosphorothioates, C3-C 10 In some embodiments, L is an alkenyl-phosphorothioate, a C3-C9 alkenyl-phosphorothioate, a C3-C8 alkenyl-phosphorothioate, a C3-C7 alkenyl-phosphorothioate, a C3-C6 alkenyl-phosphorothioate, a C3-C5 alkenyl-phosphorothioate, a C3-C4 alkenyl-phosphorothioate, or a C3 alkenyl-phosphorothioate. 4 is C3-C 20 Alkenyl-boranophosphate, C3-C 19 Alkenyl-boranophosphate, C3-C18 Alkenyl-boranophosphate, C3-C 17 Alkenyl-boranophosphate, C3-C 16 Alkenyl-boranophosphate, C3-C 15 Alkenyl-boranophosphate, C3-C 14 Alkenyl-boranophosphate, C3-C 13 Alkenyl-boranophosphate, C3-C 12 Alkenyl-boranophosphate, C3-C 11 Alkenyl-boranophosphate, C3-C 10 alkenyl-boranophosphate, C3-C9 alkenyl-boranophosphate, C3-C8 alkenyl-boranophosphate, C3-C7 alkenyl-boranophosphate, C3-C6 alkenyl-boranophosphate, C3-C5 alkenyl-boranophosphate, C3-C4 alkenyl-boranophosphate, or C3 alkenyl-boranophosphate.

[0055] In some embodiments, L 4 is -C(O)NH-C1-C 10 alkyl-phosphate, —C(O)NH—C1-C9 alkyl-phosphate, —C(O)NH—C1-C8 alkyl-phosphate, —C(O)NH—C1-C7 alkyl-phosphate, —C(O)NH—C1-C6 alkyl-phosphate, —C(O)NH—C1-C5 alkyl-phosphate, —C(O)NH—C1-C4 alkyl-phosphate, —C(O)NH—C1-C3 alkyl-phosphate, —C(O)NH—C1-C2 alkyl-phosphate, or —C(O)NH—CH2-phosphate.

[0056] In some embodiments, L 4 is -C(O)NH-C3-C 10alkenyl-phosphate, -C(O)NH-C3-C9 alkenyl-phosphate, -C(O)NH-C3-C8 alkenyl-phosphate, -C(O)NH-C3-C7 alkenyl-phosphate, -C(O)NH-C3-C6 alkenyl-phosphate, -C(O)NH-C3-C5 alkenyl-phosphate, -C(O)NH-C3-C4 alkenyl-phosphate, or -C(O)NH-C3 alkenyl-phosphate.

[0057] In some embodiments, L 4 is -C(O)O-C1-C 10 alkyl-phosphate, —C(O)O—C1-C9 alkyl-phosphate, —C(O)O—C1-C8 alkyl-phosphate, —C(O)O—C1-C7 alkyl-phosphate, —C(O)O—C1-C6 alkyl-phosphate, —C(O)O—C1-C5 alkyl-phosphate, —C(O)O—C1-C4 alkyl-phosphate, —C(O)O—C1-C3 alkyl-phosphate, —C(O)O—C1-C2 alkyl-phosphate, or —C(O)O—CH2-phosphate.

[0058] In some embodiments, L 4 is -C(O)O-C3-C 10 alkenyl-phosphate, -C(O)O-C3-C9 alkenyl-phosphate, -C(O)O-C3-C8 alkenyl-phosphate, -C(O)O-C3-C7 alkenyl-phosphate, -C(O)O-C3-C6 alkenyl-phosphate, -C(O)O-C3-C5 alkenyl-phosphate, -C(O)O-C3-C4 alkenyl-phosphate, or -C(O)O-C3 alkenyl-phosphate.

[0059] In some embodiments, L 4 is -C(O)NH-C1-C 10alkyl-phosphorothioate, -C(O)NH-C1-C9 alkyl-phosphorothioate, -C(O)NH-C1-C8 alkyl-phosphorothioate, -C(O)NH-C1-C7 alkyl-phosphorothioate, -C(O)NH-C1-C6 alkyl-phosphorothioate, -C(O)NH-C1-C5 alkyl-phosphorothioate, -C(O)NH-C1-C4 alkyl-phosphorothioate, -C(O)NH-C1-C3 alkyl-phosphorothioate, -C(O)NH-C1-C2 alkyl-phosphorothioate, or -C(O)NH-CH2-phosphorothioate.

[0060] In some embodiments, L 4 is -C(O)NH-C3-C 10 alkenyl-phosphorothioate, -C(O)NH-C3-C9 alkenyl-phosphorothioate, -C(O)NH-C3-C8 alkenyl-phosphorothioate, -C(O)NH-C3-C7 alkenyl-phosphorothioate, -C(O)NH-C3-C6 alkenyl-phosphorothioate, -C(O)NH-C3-C5 alkenyl-phosphorothioate, -C(O)NH-C3-C4 alkenyl-phosphorothioate, or -C(O)NH-C3 alkenyl-phosphorothioate.

[0061] In some embodiments, L 4 is -C(O)O-C1-C 10 alkyl-phosphorothioate, -C(O)O-C1-C9 alkyl-phosphorothioate, -C(O)O-C1-C8 alkyl-phosphorothioate, -C(O)O-C1-C7 alkyl-phosphorothioate, -C(O)O-C1-C6 alkyl-phosphorothioate, -C(O)O-C1-C5 alkyl-phosphorothioate, -C(O)O-C1-C4 alkyl-phosphorothioate, -C(O)O-C1-C3 alkyl-phosphorothioate, -C(O)O-C1-C2 alkyl-phosphorothioate, or -C(O)O-CH2-phosphorothioate.

[0062] In some embodiments, L 4is -C(O)O-C3-C 10 alkenyl-phosphorothioate, -C(O)O-C3-C9 alkenyl-phosphorothioate, -C(O)O-C3-C8 alkenyl-phosphorothioate, -C(O)O-C3-C7 alkenyl-phosphorothioate, -C(O)O-C3-C6 alkenyl-phosphorothioate, -C(O)O-C3-C5 alkenyl-phosphorothioate, -C(O)O-C3-C4 alkenyl-phosphorothioate, or -C(O)O-C3 alkenyl-phosphorothioate.

[0063] In some embodiments, L 4 is -C(O)-NH-C1-C 10 alkyl-boranophosphate, —C(O)—NH—C1-C9 alkyl-boranophosphate, —C(O)—NH—C1-C8 alkyl-boranophosphate, —C(O)—NH—C1-C7 alkyl-boranophosphate, —C(O)—NH—C1-C6 alkyl-boranophosphate, —C(O)—NH—C1-C5 alkyl-boranophosphate, —C(O)—NH—C1-C4 alkyl-boranophosphate, —C(O)—NH—C1-C3 alkyl-boranophosphate, —C(O)—NH—C1-C2 alkyl-boranophosphate, or —C(O)—NH—CH2-boranophosphate.

[0064] In some embodiments, L 4 is -C(O)-NH-C3-C 10 alkenyl-boranophosphate, -C(O)-NH-C3-C9 alkenyl-boranophosphate, -C(O)-NH-C3-C8 alkenyl-boranophosphate, -C(O)-NH-C3-C7 alkenyl-boranophosphate, -C(O)-NH-C3-C6 alkenyl-boranophosphate, -C(O)-NH-C3-C5 alkenyl-boranophosphate, -C(O)-NH-C3-C4 alkenyl-boranophosphate, or -C(O)-NH-C3 alkenyl-boranophosphate.

[0065] In some embodiments, L 4is -C(O)O-C1-C 10 alkyl-boranophosphate, —C(O)O—C1-C9 alkyl-boranophosphate, —C(O)O—C1-C8 alkyl-boranophosphate, —C(O)O—C1-C7 alkyl-boranophosphate, —C(O)O—C1-C6 alkyl-boranophosphate, —C(O)O—C1-C5 alkyl-boranophosphate, —C(O)O—C1-C4 alkyl-boranophosphate, —C(O)O—C1-C3 alkyl-boranophosphate, —C(O)O—C1-C2 alkyl-boranophosphate, or —C(O)O—CH2-boranophosphate.

[0066] In some embodiments, L 4 is -C(O)O-C3-C 10 alkenyl-boranophosphate, —C(O)O—C3-C9 alkenyl-boranophosphate, —C(O)O—C3-C8 alkenyl-boranophosphate, —C(O)O—C3-C7 alkenyl-boranophosphate, —C(O)O—C3-C6 alkenyl-boranophosphate, —C(O)O—C3-C5 alkenyl-boranophosphate, —C(O)O—C3-C4 alkenyl-boranophosphate, or —C(O)O—C3 alkenyl-boranophosphate.

[0067] In some embodiments, R 1 is selected from the group consisting of pentafluorophenyl, tetrafluorophenyl, succinimide, maleimide, azide, pyridyldithiol, dimethyl methylphosphonate, chiral dimethyl methylphosphonate, helper lipids, and nucleic acids. 1 is an ASO (antisense oligonucleotide), siRNA (small interfering RNA), miRNA (microRNA), microRNA mimic, AMO (anti-miRNA oligonucleotide), long non-coding RNA, PNA (peptide nucleic acid), helper lipid, or PMO (phosphorodiamidate morpholino oligomer), wherein the nucleic acid is optionally modified. In some embodiments, R 1is an ASO (antisense oligonucleotide).

[0068] In another embodiment, disclosed herein is a compound of formula IC [ka] or a pharmaceutically acceptable salt or solvate thereof, wherein X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n wherein n is 1 to 36, m is 1 to 30, and R N is H, methyl, or CH2F, CHF2, or CF3; Y 1 , Y 2 and Y 3 are each independently selected from the group consisting of -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, and P(Z)(OH)O2, where Z is O or S; L 1 , L 2 and L 3 are each independently C1-C 10 Alkyl, -(CH2) e -O-(CH2) f -, -(CH2) e -S-(CH2) f -, -(CH2) e -S(O)2-(CH2) f -, -(CH2) e -NR N -(CH2) f - and -(CH2-CH2-O) k (CH2)2-, wherein e is 1 to 10, f is 1 to 16, k is 1 to 20, and R N is H, methyl, or CH2F, CHF2, or CF3; G 1 , G2 and G 3 are each independently selected from the group consisting of monosaccharides, monosaccharide derivatives, vitamins, polyols, polysialic acids, and polysialic acid derivatives; 4 is (a)-(CH2) g -O-(CH2) h -or-(CH2) g -NR N -(CH2) h wherein g is 1 to 30, h is 1 to 36, and R N is H, methyl, or CHF, CHF, or CF), (b) amino acids, and (c) -NHC(O)R 2 (In the formula, R 2 is C1-C 10 Alkyl, carbocycle, heterocyclyl, heteroaryl, C1-C 10 Alkyl-Carbocyclic, C1-C 10 Alkyl-heterocyclyl or C1-C 10 alkyl-heteroaryl, and wherein R 2 is selected from the group consisting of: [ka] [ka] where H 1 is carbocycle, heterocyclyl, or heteroaryl; H 1 is optionally substituted;W 1 and W 2 are each independently selected from —CH— and O; v is 1 to 6; wherein Y is hydrogen or methyl; and T is C-C 10 Alkyl or C1-C 10 Alkenyl; L 4 -C(O)O-, -C(O)NH-, phosphate, C1-C 10 Alkyl-phosphate, C3-C 10 Alkenyl-phosphate, phosphorothioate, C1-C 10 Alkyl-phosphorothioates, C3-C 10Alkenyl-phosphorothioates, Boranophosphates, C1-C 10 Alkyl-boranophosphate, C3-C 10 Alkenyl-boranophosphate, -C(O)NH-C-C 10 Alkyl-phosphate, -C(O)NH-C3-C 10 Alkenyl phosphate, -C(O)O-C1-C 10 Alkyl-phosphate, -C(O)O-C3-C 10 Alkenyl-phosphate, -C(O)NH-C1-C 10 Alkyl-phosphorothioate, -C(O)NH-C3-C 10 Alkenyl-phosphorothioate, -C(O)O-C1-C 10 Alkyl-phosphorothioate, -C(O)O-C3-C 10 Alkenyl-phosphorothioate, -C(O)-NH-C-C 10 Alkyl-boranophosphate, -C(O)-NH-C3-C 10 Alkenyl-boranophosphate, -C(O)O-C1-C 10 Alkyl-boranophosphate or -C(O)O-C3-C 10 alkenyl-boranophosphate; and R 1 is a bioactive molecule.

[0069] In some embodiments, W 1 and W 2 are each independently selected from —CH2— and O, wherein v is 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or —CH2—.

[0070] In some embodiments, T is C1-C 10 In some embodiments, T is C2-C3 alkyl, C1-C4 alkyl, C1-C5 alkyl, C1-C6 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. 10alkenyl, C2-C9 alkenyl, C2-C8 alkenyl, C2-C7 alkenyl, C2-C6 alkenyl, C2-C5 alkenyl, C2-C4 alkenyl, C2-C3 alkenyl or -CH=CH-.

[0071] In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n -, where n is 1 to 36, 1 to 35, 1 to 34, 1 to 33, 1 to 32, 1 to 31, 1 to 30, 1 to 29, 1 to 28, 1 to 27, 1 to 26, 1 to 25, 1 to 24, 1 to 23, 1 to 22, 1 to 21, 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, and 1. In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n -, wherein m is 1 to 30, 1 to 29, 1 to 28, 1 to 27, 1 to 26, 1 to 25, 1 to 24, 1 to 23, 1 to 22, 1 to 21, 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, and 1. In some embodiments, X 1 , X 2 and X 3 are each independently (-CH2) m -O-CH2-, where m is 1 to 4. In some embodiments, X 1 , X2 and X 3 are each independently (—CH)—O—CH—. In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl or -CH2-.

[0072] In some embodiments, Y 1 , Y 2 and Y 3 are each —NHC(O)— or —C(O)NH—. In some embodiments, Y 1 , Y 2 and Y 3 are each -NHC(O)-.

[0073] In some embodiments, L 1 , L 2 and L 3 are C1-C 10 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. In some embodiments, L 1 , L 2 and L 3 are each independently C3-C8 alkyl or -(CH2-CH2-O) k (CH2)2-, where k is 1 to 10. In some embodiments, L 1 , L 2 and L 3 are each independently -(CH2-CH2-O) k (CH2)2-, where k is 2 to 4. In some embodiments, L 1 , L 2 and L 3 are each independently -(CH2-CH2-O) k(CH2)2-, where k is 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1. In some embodiments, L 1 , L 2 and L 3 are each independently —(CH—CH—O)(CH)—.

[0074] In some embodiments, G 1 , G 2 and G 3 are each independently folic acid, ribose, retinol, niacin, riboflavin, biotin, glucose, mannose, fucose, sucrose, lactose, mannose-6-phosphate, N-acetylgalactosamine, N-acetylglucosamine, sialic acid, sialic acid derivatives, allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, fucitol, fucosamine, fucose, fuculose, galactosamine, galactosaminitol, galactose, glucosamine, glucosaminitol, glucose-6 In some embodiments, the sugar is selected from the group consisting of phosphate, guloseglyceraldehyde, glycero-mannoseptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, psicose, quinovose, quinovosamine, rhamnitol, rhamnosamine, rhamnose, ribulose, sedoheptulose, sorbose, tagatose, talose, threose, xylose, and xylulose. 1 , G 2 and G 3 are each N-acetylgalactosamine.

[0075] In some embodiments, X 4 teeth, [ka] wherein X 4 is optionally substituted.

[0076] In some embodiments, X 4 is -NHC(O)R2 where R 2 is a carbocycle, heterocyclyl, or heteroaryl, where R 2 is optionally substituted. In some embodiments, X 4 is -NHC(O)R 2 where R 2 is a carbocycle, heterocyclyl, or heteroaryl, where R 2 is optionally substituted with alkyl, alkoxy, or amine. In some embodiments, X 4 teeth, [ka] is.

[0077] In some embodiments, L 4 -C(O)O-, -C(O)NH-, phosphate, C1-C 10 Alkyl-phosphate, C3-C 10 Alkenyl-phosphate, phosphorothioate, C1-C 10 Alkyl-phosphorothioates, C3-C 10 Alkenyl-phosphorothioates, Boranophosphates, C1-C 10 Alkyl-boranophosphate, C3-C 10 Alkenyl-boranophosphate, -C(O)NH-C-C 10 Alkyl-phosphate, -C(O)NH-C3-C 10 Alkenyl phosphate, -C(O)O-C1-C 10 Alkyl-phosphate, -C(O)O-C3-C 10 Alkenyl-phosphate, -C(O)NH-C1-C 10 Alkyl-phosphorothioate, -C(O)NH-C3-C 10 Alkenyl-phosphorothioate, -C(O)O-C1-C 10 Alkyl-phosphorothioate, -C(O)O-C3-C 10 Alkenyl-phosphorothioate, -C(O)-NH-C-C 10 Alkyl-boranophosphate, -C(O)-NH-C3-C10 Alkenyl-boranophosphate, -C(O)O-C1-C 10 Alkyl-boranophosphate or -C(O)O-C3-C 10 It is an alkenyl-boranophosphate.

[0078] In some embodiments, L 4 is C1-C 20 Alkyl-phosphates, C1-C 19 Alkyl-phosphates, C1-C 18 Alkyl-phosphates, C1-C 17 Alkyl-phosphates, C1-C 16 Alkyl-phosphates, C1-C 15 Alkyl-phosphates, C1-C 14 Alkyl-phosphates, C1-C 13 Alkyl-phosphates, C1-C 12 Alkyl-phosphates, C1-C 11 Alkyl-phosphates, C1-C 10 alkyl-phosphate, C1-C9 alkyl-phosphate, C1-C8 alkyl-phosphate, C1-C7 alkyl-phosphate, C1-C6 alkyl-phosphate, C1-C5 alkyl-phosphate, C1-C4 alkyl-phosphate, C1-C3 alkyl-phosphate, C1-C2 alkyl-phosphate, or -CH2-phosphate. 4 is C1-C 20 Alkyl-phosphorothioates, C1-C 19 Alkyl-phosphorothioates, C1-C 18 Alkyl-phosphorothioates, C1-C 17 Alkyl-phosphorothioates, C1-C 16 Alkyl-phosphorothioates, C1-C 15 Alkyl-phosphorothioates, C1-C 14 Alkyl-phosphorothioates, C1-C 13 Alkyl-phosphorothioates, C1-C 12 Alkyl-phosphorothioates, C1-C 11Alkyl-phosphorothioates, C1-C 10 alkyl-phosphorothioate, C1-C9 alkyl-phosphorothioate, C1-C8 alkyl-phosphorothioate, C1-C7 alkyl-phosphorothioate, C1-C6 alkyl-phosphorothioate, C1-C5 alkyl-phosphorothioate, C1-C4 alkyl-phosphorothioate, C1-C3 alkyl-phosphorothioate, C1-C2 alkyl-phosphorothioate or -CH2-phosphorothioate. 4 is C1-C 20 Alkyl-boranophosphates, C1-C 19 Alkyl-boranophosphates, C1-C 18 Alkyl-boranophosphates, C1-C 17 Alkyl-boranophosphates, C1-C 16 Alkyl-boranophosphates, C1-C 15 Alkyl-boranophosphates, C1-C 14 Alkyl-boranophosphates, C1-C 13 Alkyl-boranophosphates, C1-C 12 Alkyl-boranophosphates, C1-C 11 Alkyl-boranophosphates, C1-C 10 alkyl-boranophosphate, C1-C9 alkyl-boranophosphate, C1-C8 alkyl-boranophosphate, C1-C7 alkyl-boranophosphate, C1-C6 alkyl-boranophosphate, C1-C5 alkyl-boranophosphate, C1-C4 alkyl-boranophosphate, C1-C3 alkyl-boranophosphate, C1-C2 alkyl-boranophosphate or -CH2-boranophosphate.

[0079] In some embodiments, L 4 is C3-C 20 Alkenyl Phosphates, C3-C 19 Alkenyl Phosphates, C3-C 18 Alkenyl Phosphates, C3-C 17 Alkenyl Phosphates, C3-C 16Alkenyl Phosphates, C3-C 15 Alkenyl Phosphates, C3-C 14 Alkenyl Phosphates, C3-C 13 Alkenyl Phosphates, C3-C 12 Alkenyl Phosphates, C3-C 11 Alkenyl Phosphates, C3-C 10 alkenyl-phosphate, C3-C9 alkenyl-phosphate, C3-C8 alkenyl-phosphate, C3-C7 alkenyl-phosphate, C3-C6 alkenyl-phosphate, C3-C5 alkenyl-phosphate, C3-C4 alkenyl-phosphate, or C3 alkenyl-phosphate. 4 is C3-C 20 Alkenyl-phosphorothioates, C3-C 19 Alkenyl-phosphorothioates, C3-C 18 Alkenyl-phosphorothioates, C3-C 17 Alkenyl-phosphorothioates, C3-C 16 Alkenyl-phosphorothioates, C3-C 15 Alkenyl-phosphorothioates, C3-C 14 Alkenyl-phosphorothioates, C3-C 13 Alkenyl-phosphorothioates, C3-C 12 Alkenyl-phosphorothioates, C3-C 11 Alkenyl-phosphorothioates, C3-C 10 In some embodiments, L is an alkenyl-phosphorothioate, a C3-C9 alkenyl-phosphorothioate, a C3-C8 alkenyl-phosphorothioate, a C3-C7 alkenyl-phosphorothioate, a C3-C6 alkenyl-phosphorothioate, a C3-C5 alkenyl-phosphorothioate, a C3-C4 alkenyl-phosphorothioate, or a C3 alkenyl-phosphorothioate. 4 is C3-C 20 Alkenyl-boranophosphate, C3-C 19 Alkenyl-boranophosphate, C3-C 18 Alkenyl-boranophosphate, C3-C17 Alkenyl-boranophosphate, C3-C 16 Alkenyl-boranophosphate, C3-C 15 Alkenyl-boranophosphate, C3-C 14 Alkenyl-boranophosphate, C3-C 13 Alkenyl-boranophosphate, C3-C 12 Alkenyl-boranophosphate, C3-C 11 Alkenyl-boranophosphate, C3-C 10 alkenyl-boranophosphate, C3-C9 alkenyl-boranophosphate, C3-C8 alkenyl-boranophosphate, C3-C7 alkenyl-boranophosphate, C3-C6 alkenyl-boranophosphate, C3-C5 alkenyl-boranophosphate, C3-C4 alkenyl-boranophosphate, or C3 alkenyl-boranophosphate.

[0080] In some embodiments, L 4 is -C(O)NH-C1-C 10 alkyl-phosphate, —C(O)NH—C1-C9 alkyl-phosphate, —C(O)NH—C1-C8 alkyl-phosphate, —C(O)NH—C1-C7 alkyl-phosphate, —C(O)NH—C1-C6 alkyl-phosphate, —C(O)NH—C1-C5 alkyl-phosphate, —C(O)NH—C1-C4 alkyl-phosphate, —C(O)NH—C1-C3 alkyl-phosphate, —C(O)NH—C1-C2 alkyl-phosphate, or —C(O)NH—CH2-phosphate.

[0081] In some embodiments, L 4 is -C(O)NH-C3-C 10alkenyl-phosphate, -C(O)NH-C3-C9 alkenyl-phosphate, -C(O)NH-C3-C8 alkenyl-phosphate, -C(O)NH-C3-C7 alkenyl-phosphate, -C(O)NH-C3-C6 alkenyl-phosphate, -C(O)NH-C3-C5 alkenyl-phosphate, -C(O)NH-C3-C4 alkenyl-phosphate, or -C(O)NH-C3 alkenyl-phosphate.

[0082] In some embodiments, L 4 is -C(O)O-C1-C 10 alkyl-phosphate, —C(O)O—C1-C9 alkyl-phosphate, —C(O)O—C1-C8 alkyl-phosphate, —C(O)O—C1-C7 alkyl-phosphate, —C(O)O—C1-C6 alkyl-phosphate, —C(O)O—C1-C5 alkyl-phosphate, —C(O)O—C1-C4 alkyl-phosphate, —C(O)O—C1-C3 alkyl-phosphate, —C(O)O—C1-C2 alkyl-phosphate, or —C(O)O—CH2-phosphate.

[0083] In some embodiments, L 4 is -C(O)O-C3-C 10 alkenyl-phosphate, -C(O)O-C3-C9 alkenyl-phosphate, -C(O)O-C3-C8 alkenyl-phosphate, -C(O)O-C3-C7 alkenyl-phosphate, -C(O)O-C3-C6 alkenyl-phosphate, -C(O)O-C3-C5 alkenyl-phosphate, -C(O)O-C3-C4 alkenyl-phosphate, or -C(O)O-C3 alkenyl-phosphate.

[0084] In some embodiments, L 4 is -C(O)NH-C1-C 10alkyl-phosphorothioate, -C(O)NH-C1-C9 alkyl-phosphorothioate, -C(O)NH-C1-C8 alkyl-phosphorothioate, -C(O)NH-C1-C7 alkyl-phosphorothioate, -C(O)NH-C1-C6 alkyl-phosphorothioate, -C(O)NH-C1-C5 alkyl-phosphorothioate, -C(O)NH-C1-C4 alkyl-phosphorothioate, -C(O)NH-C1-C3 alkyl-phosphorothioate, -C(O)NH-C1-C2 alkyl-phosphorothioate, or -C(O)NH-CH2-phosphorothioate.

[0085] In some embodiments, L 4 is -C(O)NH-C3-C 10 alkenyl-phosphorothioate, -C(O)NH-C3-C9 alkenyl-phosphorothioate, -C(O)NH-C3-C8 alkenyl-phosphorothioate, -C(O)NH-C3-C7 alkenyl-phosphorothioate, -C(O)NH-C3-C6 alkenyl-phosphorothioate, -C(O)NH-C3-C5 alkenyl-phosphorothioate, -C(O)NH-C3-C4 alkenyl-phosphorothioate, or -C(O)NH-C3 alkenyl-phosphorothioate.

[0086] In some embodiments, L 4 is -C(O)O-C1-C 10 alkyl-phosphorothioate, -C(O)O-C1-C9 alkyl-phosphorothioate, -C(O)O-C1-C8 alkyl-phosphorothioate, -C(O)O-C1-C7 alkyl-phosphorothioate, -C(O)O-C1-C6 alkyl-phosphorothioate, -C(O)O-C1-C5 alkyl-phosphorothioate, -C(O)O-C1-C4 alkyl-phosphorothioate, -C(O)O-C1-C3 alkyl-phosphorothioate, -C(O)O-C1-C2 alkyl-phosphorothioate, or -C(O)O-CH2-phosphorothioate.

[0087] In some embodiments, L 4is -C(O)O-C3-C 10 alkenyl-phosphorothioate, -C(O)O-C3-C9 alkenyl-phosphorothioate, -C(O)O-C3-C8 alkenyl-phosphorothioate, -C(O)O-C3-C7 alkenyl-phosphorothioate, -C(O)O-C3-C6 alkenyl-phosphorothioate, -C(O)O-C3-C5 alkenyl-phosphorothioate, -C(O)O-C3-C4 alkenyl-phosphorothioate, or -C(O)O-C3 alkenyl-phosphorothioate.

[0088] In some embodiments, L 4 is -C(O)-NH-C1-C 10 alkyl-boranophosphate, —C(O)—NH—C1-C9 alkyl-boranophosphate, —C(O)—NH—C1-C8 alkyl-boranophosphate, —C(O)—NH—C1-C7 alkyl-boranophosphate, —C(O)—NH—C1-C6 alkyl-boranophosphate, —C(O)—NH—C1-C5 alkyl-boranophosphate, —C(O)—NH—C1-C4 alkyl-boranophosphate, —C(O)—NH—C1-C3 alkyl-boranophosphate, —C(O)—NH—C1-C2 alkyl-boranophosphate, or —C(O)—NH—CH2-boranophosphate.

[0089] In some embodiments, L 4 is -C(O)-NH-C3-C 10 alkenyl-boranophosphate, -C(O)-NH-C3-C9 alkenyl-boranophosphate, -C(O)-NH-C3-C8 alkenyl-boranophosphate, -C(O)-NH-C3-C7 alkenyl-boranophosphate, -C(O)-NH-C3-C6 alkenyl-boranophosphate, -C(O)-NH-C3-C5 alkenyl-boranophosphate, -C(O)-NH-C3-C4 alkenyl-boranophosphate, or -C(O)-NH-C3 alkenyl-boranophosphate.

[0090] In some embodiments, L 4is -C(O)O-C1-C 10 alkyl-boranophosphate, —C(O)O—C1-C9 alkyl-boranophosphate, —C(O)O—C1-C8 alkyl-boranophosphate, —C(O)O—C1-C7 alkyl-boranophosphate, —C(O)O—C1-C6 alkyl-boranophosphate, —C(O)O—C1-C5 alkyl-boranophosphate, —C(O)O—C1-C4 alkyl-boranophosphate, —C(O)O—C1-C3 alkyl-boranophosphate, —C(O)O—C1-C2 alkyl-boranophosphate, or —C(O)O—CH2-boranophosphate.

[0091] In some embodiments, L 4 is -C(O)O-C3-C 10 alkenyl-boranophosphate, —C(O)O—C3-C9 alkenyl-boranophosphate, —C(O)O—C3-C8 alkenyl-boranophosphate, —C(O)O—C3-C7 alkenyl-boranophosphate, —C(O)O—C3-C6 alkenyl-boranophosphate, —C(O)O—C3-C5 alkenyl-boranophosphate, —C(O)O—C3-C4 alkenyl-boranophosphate, or —C(O)O—C3 alkenyl-boranophosphate.

[0092] In some embodiments, R 1 is selected from the group consisting of pentafluorophenyl, tetrafluorophenyl, succinimide, maleimide, azide, pyridyldithiol, dimethyl methylphosphonate, chiral dimethyl methylphosphonate, helper lipids, and nucleic acids. 1 is an ASO (antisense oligonucleotide), siRNA (small interfering RNA), miRNA (microRNA), microRNA mimic, AMO (anti-miRNA oligonucleotide), long non-coding RNA, PNA (peptide nucleic acid), helper lipid, or PMO (phosphorodiamidate morpholino oligomer), wherein the nucleic acid is optionally modified. In some embodiments, R 1is an ASO (antisense oligonucleotide).

[0093] In some embodiments, the compound has the formula [ka] [ka] [ka] and During the ceremony, [ka] is an oligonucleotide, and R p is C1-C 10 Alkyl or C2-C 10 In some embodiments, R is alkenyl. p is C1-C 10 Alkyl or C2-C 10 alkenyl, C1-C9 alkyl or C2-C9 alkenyl, C1-C8 alkyl or C2-C8 alkenyl, C1-C7 alkyl or C2-C7 alkenyl, C1-C6 alkyl or C2-C6 alkenyl, C1-C5 alkyl or C2-C5 alkenyl, C1-C4 alkyl or C2-C4 alkenyl, C1-C3 alkyl or C2-C3 alkenyl, C1-C2 alkyl, -CH=CH- or -CH2-.

[0094] In some embodiments, the compound is [ka] [ka] [ka] [ka] and During the ceremony, [ka] is an oligonucleotide.

[0095] In some embodiments, compounds comprising Formula B, Formula C, Formula D, Formula E, and Formula F are each independently derived from a click chemistry process.

[0096] In another embodiment, disclosed herein is a compound of formula IA, formula IB, or formula IC, and a lipid of formula II [ka] or a pharmaceutically acceptable salt or solvate thereof, wherein R 5 and R 6 are each independently a straight-chain or branched C1-C 31 Alkyl, C2-C 31 Alkenyl or C2-C 31 selected from the group consisting of alkynyl and cholesteryl; L 5 and L 6 are each independently a straight chain C1-C 20 Alkyl and C1-C 20 alkenyl; X 5 is -C(O)O- or -OC(O)-; X 6 are -C(O)O- and -OC(O)-; X 7 is S or O; L 7 is absent or lower alkyl; R 4 is a straight-chain or branched C1-C6 alkyl; and R 7 and R 8 are each independently selected from the group consisting of hydrogen and straight-chain or branched C1-C6 alkyl.

[0097] In some embodiments, X 7 is S.

[0098] In some embodiments, R 7 and R 8are each independently selected from the group consisting of methyl, ethyl, and isopropyl. 7 and R 8 are each independently selected from the group consisting of propyl, butyl, isobutyl, t-butyl, pentyl, and hexyl.

[0099] In some embodiments, L 5 and L 6 are each independently C1-C 10 In some embodiments, L 5 and L 6 are each independently C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. 5 is C1-C3 alkyl, and L 6 is C1-C5 alkyl. In some embodiments, L 6 is C1-C2 alkyl. In some embodiments, L 5 and L 6 Each is a straight chain C7 alkyl. In some embodiments, L 5 and L 6 are each a straight chain C9 alkyl.

[0100] In some embodiments, R 5 and R 6 are each independently a straight-chain or branched C1-C 31 Alkyl, C1-C 30 Alkyl, C1-C 29 Alkyl, C1-C 28 Alkyl, C1-C 27 Alkyl, C1-C 26 Alkyl, C1-C 25 Alkyl, C1-C 24 Alkyl, C1-C 23 Alkyl, C1-C 22 Alkyl, C1-C 21 Alkyl, C1-C 20 Alkyl, C1-C 19 Alkyl, C1-C18 Alkyl, C1-C 17 Alkyl, C1-C 16 Alkyl, C1-C 15 Alkyl, C1-C 14 Alkyl, C1-C 13 Alkyl, C1-C 12 Alkyl, C1-C 11 Alkyl, C1-C 10 It is selected from the group consisting of alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, and —CH2—.

[0101] In some embodiments, R 5 and R 6 are each independently a straight-chain or branched C-C 31 Alkenyl, C2-C 30 Alkenyl, C2-C 29 Alkenyl, C2-C 28 Alkenyl, C2-C 27 Alkenyl, C2-C 26 Alkenyl, C2-C 25 Alkenyl, C2-C 24 Alkenyl, C2-C 23 Alkenyl, C2-C 22 Alkenyl, C2-C 21 Alkenyl, C2-C 20 Alkenyl, C2-C 19 Alkenyl, C2-C 18 Alkenyl, C2-C 17 Alkenyl, C2-C 16 Alkenyl, C2-C 15 Alkenyl, C2-C 14 Alkenyl, C2-C 13 Alkenyl, C2-C 12 Alkenyl, C2-C 11 Alkenyl, C2-C 10In some embodiments, R is selected from the group consisting of alkenyl, C2-C9 alkenyl, C2-C8 alkenyl, C2-C7 alkenyl, C2-C6 alkenyl, C2-C5 alkenyl, C2-C4 alkenyl, C2-C3 alkenyl, and -CH=CH-. 5 and R 6 are each independently alkenyl. In some embodiments, R 6 is alkenyl. In some embodiments, R 6 is a C2-C9 alkenyl. In some embodiments, the alkenyl contains a single double bond, two double bonds, or three double bonds. In some embodiments, R 5 and R 6 Each is alkyl. In some embodiments, R 5 is a branched alkane. In some embodiments, R 5 and R 6 are each independently selected from the group consisting of C alkyl, C alkenyl, and C alkynyl. In some embodiments, R 5 and R 6 are each independently, C 11 Alkyl, C 11 Alkenyl and C 11 In some embodiments, R is selected from the group consisting of alkynyl. 5 and R 6 are each independently selected from the group consisting of C alkyl, C alkenyl, and C alkynyl. 5 is -CH((CH2) p CH3)2 or -CH((CH2) p CH3)((CH2) p-1 CH3), where p is 4 to 8. In some embodiments, p is 5 and L 5 is C1-C3 alkyl. In some embodiments, p is 6 and L 5 is C alkyl. In some embodiments, p is 7. In some embodiments, p is 8 and L 5 is C1-C3 alkyl. In some embodiments, R 5is -CH((CH2) p CH3)((CH2) p-1 CH3), where p is 7 or 8.

[0102] In some embodiments, R 4 is ethylene or propylene. In some embodiments, R 4 is n-propylene or isobutylene.

[0103] In some embodiments, L 7 is absent, R 4 is ethylene and X 7 is S and R 7 and R 8 Each is methyl. In some embodiments, L 7 is absent, R 4 is n-propylene, and X 7 is S and R 7 and R 8 Each is methyl. In some embodiments, L 7 is absent, R 4 is ethylene and X 7 is S and R 7 and R 8 are each ethyl.

[0104] In another embodiment, disclosed herein is a compound of formula IA, formula IB, or formula IC, and [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] or a pharmaceutically acceptable salt or solvate thereof, comprising a lipid selected from the group consisting of:

[0105] In another embodiment, disclosed herein is a compound of formula IA, formula IB, or formula IC, and a lipid of formula III [ka] or a pharmaceutically acceptable salt or solvate thereof, wherein R 9 and R 10 are each independently a straight-chain or branched C 1-20 Alkyl, straight or branched C2-C 20 Alkenyl and C2-C 20 alkynyl; L 8 and L 9 are each independently absent, straight chain C1-C 18 Alkyl, or straight chain C2-C 18 alkenyl; L 11 is absent, a bond, or a straight-chain or branched C1-C6 alkyl; L 10 is absent or methyl; X 8 is S or O; R 11 is a straight-chain or branched C1-C6 alkyl; and R 12 and R 13 are each independently selected from the group consisting of hydrogen, and straight-chain and branched C1-C6 alkyl.

[0106] In some embodiments, R 9 and R 10 are each independently a straight chain alkyl or alkenyl. In some embodiments, R 9 and R 10 are each independently a straight-chain or branched C1-C 20Alkyl, C1-C 19 Alkyl, C1-C 18 Alkyl, C1-C 17 Alkyl, C1-C 16 Alkyl, C1-C 15 Alkyl, C1-C 14 Alkyl, C1-C 13 Alkyl, C1-C 12 Alkyl, C1-C 11 Alkyl, C1-C 10 In some embodiments, R is selected from the group consisting of alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, and -CH2-. 9 and R 10 are each independently a straight-chain or branched C-C 20 Alkenyl, C2-C 19 Alkenyl, C2-C 18 Alkenyl, C2-C 17 Alkenyl, C2-C 16 Alkenyl, C2-C 15 Alkenyl, C2-C 14 Alkenyl, C2-C 13 Alkenyl, C2-C 12 Alkenyl, C2-C 11 Alkenyl, C2-C 10 In some embodiments, R is selected from the group consisting of alkenyl, C2-C9 alkenyl, C2-C8 alkenyl, C2-C7 alkenyl, C2-C6 alkenyl, C2-C5 alkenyl, C2-C4 alkenyl, C2-C3 alkenyl, and -CH=CH-. 9 and R 10 are each independently a straight-chain or branched C-C 20 Alkynyl, C2-C 19 Alkynyl, C2-C 18 Alkynyl, C2-C 17 Alkynyl, C2-C 16 Alkynyl, C2-C 15 Alkynyl, C2-C 14 Alkynyl, C2-C 13 Alkynyl, C2-C 12 Alkynyl, C2-C11 Alkynyl, C2-C 10 It is selected from the group consisting of alkynyl, C2-C9 alkynyl, C2-C8 alkynyl, C2-C7 alkynyl, C2-C6 alkynyl, C2-C5 alkynyl, C2-C4 alkynyl, C2-C3 alkynyl and C2 alkynyl.

[0107] In some embodiments, L 8 and L 9 are each independently a straight-chain or branched C1-C 18 Alkyl, C1-C 17 Alkyl, C1-C 16 Alkyl, C1-C 15 Alkyl, C1-C 14 Alkyl, C1-C 13 Alkyl, C1-C 12 Alkyl, C1-C 11 Alkyl, C1-C 10 In some embodiments, L is selected from the group consisting of alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, and -CH2-. 8 and L 9 are each independently a straight-chain or branched C-C 18 Alkenyl, C2-C 17 Alkenyl, C2-C 16 Alkenyl, C2-C 15 Alkenyl, C2-C 14 Alkenyl, C2-C 13 Alkenyl, C2-C 12 Alkenyl, C2-C 11 Alkenyl, C2-C 10 In some embodiments, L is selected from the group consisting of alkenyl, C2-C9 alkenyl, C2-C8 alkenyl, C2-C7 alkenyl, C2-C6 alkenyl, C2-C5 alkenyl, C2-C4 alkenyl, C2-C3 alkenyl, and -CH=CH-. 8 and L 9 are each independently a straight chain alkyl. In some embodiments, L 8 and L 9are each absent.

[0108] In some embodiments, L 11 is a branched C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. In some embodiments, L 11 is a bond.

[0109] In some embodiments, L 10 is absent.

[0110] In some embodiments, X 8 is S.

[0111] In some embodiments, R 12 and R 13 are each independently a straight chain or branched C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. In some embodiments, R 12 and R 13 are each independently a straight chain alkyl.

[0112] In another embodiment, disclosed herein is a compound of formula IA, formula IB, or formula IC, and a lipid of formula IV [ka] or a pharmaceutically acceptable salt or solvate thereof, wherein X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n and -(CH2) m -NR N -(CH2) n wherein n is 1 to 36, m is 1 to 30, and R N is H, methyl, or CH2F, CHF2, or CF3; Y 1, Y 2 and Y 3 are each independently selected from the group consisting of -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, and P(Z)(OH)O2, where Z is O or S; L 1 , L 2 and L 3 are each independently C1-C 10 Alkyl, -(CH2) e -O-(CH2) f -, -(CH2) e -S-(CH2) f -, -(CH2) e -S(O)2-(CH2) f - and -(CH2) e -NR N -(CH2) f -, -(CH2-CH2-O)e(CH2)2-, wherein e is 1 to 10, f is 1 to 16, k is 1 to 20, and R N is H, methyl, or CH2F, CHF2, or CF3; G 1 , G 2 and G 3 are each independently selected from the group consisting of monosaccharides, monosaccharide derivatives, vitamins, polyols, polysialic acids, and polysialic acid derivatives; 4 is (a)-(CH2) g -O-(CH2) h -or-(CH2) g -NR N -(CH2) h wherein g is 1 to 30, h is 1 to 36, and R N is H, methyl, or CHF, CHF, or CF), (b) amino acids, and (c) -NHC(O)R 2 (In the formula, R 2 is C1-C 10 Alkyl, carbocycle, heterocyclyl, heteroaryl, C1-C 10 Alkyl-Carbocyclic, C1-C 10 Alkyl-heterocyclyl or C1-C 10 alkyl-heteroaryl, and wherein R2 is optionally substituted); Q is selected from the group consisting of alkylamino, —O(O)C—, —C(O)O—, —NHC(O)—, —C(O)NH—, —(CH) j -NHC(O)-, -C(O)NH-(CH2) j -, -C(O)-(CH2) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is 1 to 30, j is 1 to 36, and wherein R 3 is hydrogen or alkyl; L 4 -PEG-C(O)O-, PEG-C(O)NH-, -PEG-NHC(O)-, -PEG-phosphate, -PEG-C1-C 10 Alkyl-phosphate, -PEG-C3-C 10 Alkenyl-phosphate, -PEG-phosphorothioate, -PEG-C1-C 10 Alkyl-phosphorothioate, -PEG-C3-C 10 Alkenyl-phosphorothioate, -PEG-boranophosphate, -PEG-C1-C 10 Alkyl-boranophosphate, -PEG-C3-C 10 Alkenyl-boranophosphate, -PEG-C(O)NH-C-C 10Alkyl-phosphate, -PEG-C(O)NH-C3-C 10 Alkenyl-phosphate, -PEG-C(O)O-C1-C 10 Alkyl-phosphate, -PEG-C(O)O-C3-C 10 Alkenyl-phosphate, -PEG-C(O)NH-C1-C 10 Alkyl-phosphorothioate, -PEG-C(O)NH-C3-C 10 Alkenyl-phosphorothioate, -PEG-C(O)O-C1-C 10 Alkyl-phosphorothioate, -PEG-C(O)O-C3-C 10 Alkenyl-phosphorothioate, -PEG-C(O)-NH-C-C 10 Alkyl-boranophosphate, -PEG-C(O)-NH-C3-C 10 Alkenyl-boranophosphate, -PEG-C(O)O-C1-C 10 Alkyl-boranophosphate or -PEG-C(O)O-C3-C 10 alkenyl-boranophosphate; and R 1 is a helper lipid.

[0113] In some embodiments, L 4 PEG is -(CH2-CH2-O) k(CH2)2-, wherein k is 10 to 100, 15 to 100, 10 to 90, 15 to 90, 10 to 80, 15 to 80, 10 to 70, 15 to 70, 10 to 60, 15 to 60, 10 to 50, 15 to 50, 10 to 40, 15 to 40, 10 to 30, 15 to 30, 10 to 20, 15 to 20, 20 to 90, 25 to 90, 20 to 80, 25 to 80, 20 to 70, 25 to 70, 20 to 60, 25 to 60, 20 to 50, 25 to 50, 20 to 40 , 25 to 40, 20 to 30, 25 to 30, 30 to 80, 35 to 80, 30 to 70, 35 to 70, 30 to 60, 35 to 60, 30 to 50, 35 to 50, 30 to 40, 35 to 40, 40 to 90, 45 to 90, 40 to 80, 45 to 80, 40 to 70, 45 to 70, 40 to 60, 45 to 60, 40 to 50, 45 to 50, 50 to 90, 55 to 90, 50 to 80, 55 to 80, 50 to 70, 55 to 70, 50 to 60, or 55 to 60. 4 PEG is -(CH2-CH2-O) k (CH2)2-, where k is 10 to 100, 20 to 60, 30 to 60, 40 to 60, 40 to 50, or 45 to 50. In some embodiments, L 4 PEG is -(CH2-CH2-O) k (CH2)2-, where k is 20 to 60.

[0114] In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n-, where n is 1 to 36, 1 to 35, 1 to 34, 1 to 33, 1 to 32, 1 to 31, 1 to 30, 1 to 29, 1 to 28, 1 to 27, 1 to 26, 1 to 25, 1 to 24, 1 to 23, 1 to 22, 1 to 21, 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, and 1. In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 Alkyl, -(CH2) m -O-(CH2) n - and -(CH2) m -NR N -(CH2) n -, wherein m is 1 to 30, 1 to 29, 1 to 28, 1 to 27, 1 to 26, 1 to 25, 1 to 24, 1 to 23, 1 to 22, 1 to 21, 1 to 20, 1 to 19, 1 to 18, 1 to 17, 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, and 1. In some embodiments, X 1 , X 2 and X 3 are each independently (-CH2) m -O-CH2-, where m is 1 to 4. In some embodiments, X 1 , X 2 and X 3 are each independently (—CH)—O—CH—. In some embodiments, X 1 , X 2 and X 3 are each independently C1-C 10 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. In some embodiments, X 1 , X 2 and X 3 are each independently (-CH2) m-O-CH2-, where m is 1 to 4. In some embodiments, X 1 , X 2 and X 3 are each independently (-CH2)2-O-CH2-.

[0115] In some embodiments, Y 1 , Y 2 and Y 3 are each —NHC(O)— or —C(O)NH—. In some embodiments, Y 1 , Y 2 and Y 3 are each -NHC(O)-.

[0116] In some embodiments, L 1 , L 2 and L 3 are C1-C 10 alkyl, C1-C9 alkyl, C1-C8 alkyl, C1-C7 alkyl, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, C1-C2 alkyl, or -CH2-. In some embodiments, L 1 , L 2 and L 3 are each independently C3-C8 alkyl or -(CH2-CH2-O) k (CH2)2-, where k is 1 to 10. In some embodiments, L 1 , L 2 and L 3 are each independently -(CH2-CH2-O) k (CH2)2-, where k is 2 to 4. In some embodiments, L 1 , L 2 and L 3 are each independently -(CH2-CH2-O) k (CH2)2-, where k is 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1. In some embodiments, L 1 , L 2 and L 3 are each independently —(CH—CH—O)(CH)—.

[0117] In some embodiments, G 1 , G 2 and G 3 are each independently folic acid, ribose, retinol, niacin, riboflavin, biotin, glucose, mannose, fucose, sucrose, lactose, mannose-6-phosphate, N-acetylgalactosamine, N-acetylglucosamine, sialic acid, sialic acid derivatives, allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, fucitol, fucosamine, fucose, fuculose, galactosamine, galactosaminitol, galactose, glucosamine, glucosaminitol, glucose-6 phosphate, guloseglyceraldehyde, glycero-mannoseptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, psicose, quinovose, quinovosamine, rhamnitol, rhamnosamine, rhamnose, ribulose, sedoheptulose, sorbose, tagatose, talose, threose, xylose and xylulose.

[0118] In some embodiments, G 1 , G 2 and G 3 are each N-acetylgalactosamine.

[0119] In some embodiments, X 4 teeth, [ka] wherein X 4 is optionally substituted.

[0120] In some embodiments, X 4 teeth, [ka] is.

[0121] In some embodiments, Q is alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is 1 to 10, j is 1 to 10, and wherein R 3 is hydrogen or alkyl. In some embodiments, Q is alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j-, wherein i is 1-10 and j is 1-10; i is 1-9 and j is 1-9; i is 1-8 and j is 1-8; i is 1-7 and j is 1-7; i is 1-6 and j is 1-6; i is 1-5 and j is 1-5; i is 1-5 and j is 1-4; i is 1-3 and j is 1-3; i is 1-2 and j is 1-2; or i is 1 and j is 1. In some embodiments, Q is alkylamino, -C(O)-(CH) i -, -(CH2) i -O-(CH2) j -, -(CH2) i -NR 3 -(CH2) j -, -(CH2) i -SS-(CH2) j -, -(CH2) i -S-(CH2) j -, -(CH2) i -S(O)2-(CH2) j -, -(CH2) i -NHC(O)-(CH2) j -, -(CH2) i -C(O)NH-(CH2) j -, -(CH2) i -SC(O)-(CH2) j - or -(CH2) i -C(O)S-(CH2) j -, wherein i is independently 1 to 10 and j is independently 1 to 10.

[0122] In some embodiments, R 1Lecithin, dialkyloxypropyl (DAA), diacylglycerol (DAG), dimyristoylglycerol (DMG), dioleoylglycerol (DOG), dipalmitoylglycerol (DPG), phosphatidylethanolamine (PE), distearoylglycerol (DSG), lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg yolk sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebrosides, dicetyl phosphate, distearoyl Glycerophosphocholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoyl-phosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), palmitoyloleoyl-phosphatidylglycerol (POPG), dioleoylphosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl-phosphatidylethanolamine (DPPE), dimyristoyl-phosphatidylethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), monomethyl-phosphatidylethanolamine, dimethyl-phosphatidylethanolamine, dielaidoyl-phosphatidylethanolamine (DEPE), stearoyloleoyl-phosphatidylethanolamine (SOPE), lysophosphatidylcholine, and dilinoleoylphosphatidylcholine.

[0123] In some embodiments, R 1 is selected from the group consisting of dimyristoylglycerol (DMG), dioleoylglycerol (DOG), dipalmitoylglycerol (DPG), and distearoylglycerol (DSG).

[0124] In some embodiments, the pharmaceutical composition comprises a lipid-NA (nucleic acid) nanoparticle comprising a compound of Formula IA, Formula IB, or Formula IC, and a lipid of Formula II, Formula III, or Formula IV; and a cationic lipid, a non-cationic lipid, a PEG-lipid, or a helper lipid.

[0125] In some embodiments, the lipid NA nanoparticles encapsulate a compound of formula IA, formula IB, or formula IC.

[0126] In some embodiments, the lipid NA nanoparticles have a size of less than 100 nm.

[0127] In some embodiments, the cationic lipid is a phospholipid.

[0128] In some embodiments, the non-cationic lipid is cholesterol.

[0129] In some embodiments, the PEG-lipid is PEG-diacylglycerol (PEG-DAG) or PEG-dialkyloxyalkyl (PEG-DAA). In some preferred embodiments, the PEG-lipid is PEG550-PE. In some preferred embodiments, the PEG-lipid is PEG750-PE. In some preferred embodiments, the PEG-lipid is PEG2000-DMG.

[0130] In some embodiments, the nucleic acid upregulates, suppresses, decreases, reduces, downregulates, or silences expression of the target gene.

[0131] In some embodiments, the nucleic acid modulates expression of a target gene, wherein the target gene is selected from the group consisting of MUT (methylmalonic acidemia), PCCA (propionyl-CoA carboxylase subunit alpha), PCCB (propionyl-CoA carboxylase subunit beta), ASL (argininosuccinate lyase), ASS1 (argininosuccinate synthase 1), FAH (fumarylacetoacetate hydrolase), HMBS (hydroxymethylbilane synthase), ATP7B (ATPase copper transporter beta), LDLR (low density lipoprotein receptor), G6PC (glucose-6-phosphatase catalytic subunit), and AGXT (alanine-glyoxylate and serine-pyruvate aminotransferase).

[0132] In some embodiments, the nucleic acid encodes an antibody.

[0133] In some embodiments, the antibody is capable of binding to a viral particle.

[0134] In some embodiments, the nucleic acid encodes a viral protein.

[0135] In some embodiments, the nucleic acid has a nt (nucleotide) length of 200 to 1000 nt, 1000 to 5000 nt, 5000 to 10,000 nt, or 10,000 to 25,000 nt.

[0136] In some embodiments, the nucleic acid is mRNA and / or siRNA.

[0137] In some embodiments, the nucleic acid is DNA.

[0138] In some embodiments, the nucleic acid is pDNA (plasmid DNA).

[0139] In yet another embodiment, disclosed herein is a method of treating a disease in a subject, comprising administering to the subject a pharmaceutical composition comprising a compound of Formula IA, Formula IB, or Formula IC and a lipid of Formula II, Formula III, or Formula IV; and / or lipid-NA (nucleic acid) nanoparticles comprising a cationic lipid, a non-cationic lipid, a PEG-lipid, and / or a helper lipid.

[0140] In some embodiments, administration is parenteral or by intravenous injection.

[0141] In some embodiments, administration is by subcutaneous, intradermal, or intramuscular injection.

[0142] In some embodiments, the pharmaceutical composition is administered at least twice.

[0143] It is understood that various configurations of the subject technology will be readily apparent to those skilled in the art from this disclosure, and various configurations of the subject technology are shown and described herein by way of example. As will be understood, the subject technology is capable of other and different configurations, and its several details are capable of modification in various other respects, all without departing from the scope of the subject technology. Accordingly, the summary, drawings, and detailed description are to be regarded as illustrative in nature, and not restrictive.

[0144] The following detailed description is intended to be a description of various configurations of the subject technology and is not intended to represent the only configurations in which the subject technology may be practiced. The accompanying drawings are incorporated herein and constitute a part of the detailed description. The detailed description includes specific details for the purpose of providing a thorough understanding of the subject technology. However, it will be apparent to those skilled in the art that the subject technology may be practiced without these specific details. In some instances, well-known structures and components are shown in block diagram form to avoid obscuring the concepts of the subject technology. For ease of understanding, similar components are designated with the same element numbers.

[0145] Where ranges are given, the endpoints are included. Furthermore, unless otherwise indicated or otherwise apparent from the context and the understanding of one of ordinary skill in the art, it should be understood that values ​​given as ranges can be considered to be any specific value in different embodiments of the present disclosure or subranges within the stated range, to the nearest tenth of the lower limit of that range, unless the context clearly dictates otherwise.

[0146] In addition, it should be understood that any specific embodiment of the present disclosure that falls within the scope of the prior art may be explicitly excluded from any one or more claims. Such embodiments may be excluded even if the exclusion is not explicitly stated herein, because they are considered to be known to those skilled in the art. Any specific embodiment of the composition of the present disclosure (e.g., any nucleic acid or protein encoded thereby; any manufacturing method; any method of use, etc.) may be excluded from any one or more claims for any reason, whether related to the existence of prior art or not.

[0147] At various places in the present specification, substituents of compounds of the present disclosure are disclosed in groups or in ranges. It is specifically intended that the present disclosure include all individual subcombinations of the members of such groups and ranges. For example, "C 1-6 The term "alkyl" is specifically intended to individually disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl.

[0148] The phrase "at least one" refers to one or more (eg, 1-3, 1-2, or 1).

[0149] The term "composition" refers to a product containing specified ingredients in specified amounts, as well as any product that results, directly or indirectly, from the combination of specified ingredients in specified amounts.

[0150] The phrase "in combination with" refers to the administration of a compound of Formula I together with another drug in the treatment methods of the disclosure, where the compound of Formula I and the other drug are administered sequentially or simultaneously in separate dosage forms, or simultaneously in the same dosage form.

[0151] In the claims, "a," "an," and "the" refer to one or more than one unless specifically indicated to the contrary or otherwise clear from the context. A claim or description containing "or" between one or more members of a group is considered to be satisfied when one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process, unless specifically indicated to the contrary or otherwise clear from the context. The present disclosure includes embodiments in which exactly one member of a group is present in, employed in, or otherwise relevant to a given product or process. The present disclosure includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.

[0152] The term "administered in combination" or "co-administration" refers to two or more agents being administered to a subject simultaneously or within such an interval that there may be overlap in the effects of each agent on the patient. In some embodiments, the agents are administered within about 60, 30, 15, 10, 5, or 1 minute of each other. In some embodiments, the administration of the agents is spaced sufficiently close together that a combined (e.g., synergistic) effect is achieved.

[0153] The term "animal" refers to any member of the animal kingdom. In some embodiments, "animal" refers to humans at any stage of development. In some embodiments, "animal" refers to non-human animals at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, mouse, rat, rabbit, monkey, dog, cat, sheep, cow, primate, or pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, and worms. In some embodiments, the animal is a transgenic animal, a genetically engineered animal, or a clone.

[0154] The term "approximately" or "about," as applied to one or more subject values, refers to a value similar to a stated reference value. In certain embodiments, unless otherwise specified or clear from the context, the term "approximately" or "about" refers to a range of values ​​that is within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less than 1% in either direction (greater than or less than) the stated reference value (except where such value exceeds 100% of possible values).

[0155] The terms "associated with," "conjugated," "linked," "attached," and "tethered," when used in reference to two or more moieties, refer to the moieties being physically associated or connected to one another, either directly or through one or more additional moieties that act as linking agents, to form a structure that is sufficiently stable for the moieties to remain physically associated under the conditions (e.g., physiological conditions) in which the structure is used. "Association" need not be strictly by direct covalent chemical bonding. The term can also imply connectivity based on ionic or hydrogen bonding or hybridization that is sufficiently stable for the "associated" entities to remain physically associated.

[0156] The term "biodegradable" refers to being capable of being broken down into harmless products by the activity of living organisms.

[0157] The phrase "biologically active" refers to any characteristic of a substance that has activity in a biological system and / or organism. For example, a substance is considered to be biologically active if it has a biological effect on an organism when administered to that organism. In certain embodiments, a polynucleotide of the present disclosure can be considered biologically active if even a portion of the polynucleotide mimics an activity that is considered biologically active or biologically relevant.

[0158] The term "acyl" refers to a hydrogen or alkyl group (e.g., a haloalkyl group), as defined herein, attached to the parent molecular group by a carbonyl group, as defined herein, and is exemplified by formyl (i.e., a carboxaldehyde group), acetyl, trifluoroacetyl, propionyl, butanoyl, and the like. Exemplary unsubstituted acyl groups contain 1 to 7, 1 to 11, or 1 to 21 carbons. In some embodiments, the alkyl group is further substituted with 1, 2, 3, or 4 substituents, as described herein.

[0159] The term “acylamino” refers to an acyl group, as defined herein, attached to the parent molecular moiety through an amino group, as defined herein (i.e., —N(R N1 )-C(O)-R, where R is H or optionally substituted C 1-6 , C 1-10 , or C 1-20 is an alkyl group (e.g., haloalkyl), and R N1is as defined herein). Exemplary unsubstituted acylamino groups contain 1 to 41 carbons (e.g., 1 to 7, 1 to 13, 1 to 21, 2 to 7, 2 to 13, 2 to 21, or 2 to 41 carbons). In some embodiments, the alkyl group is further substituted with 1, 2, 3, or 4 substituents as described herein, and / or the amino group is selected from -NH2 or -NHR N1 and R in the formula N1 are independently OH, NO2, NH2, and NR N2 2. SO2OR N2 , SO2R N2 , SOR N2 , alkyl, aryl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), or alkoxycarbonylalkyl, and each R N2 can be H, alkyl, or aryl.

[0160] The term “acylaminoalkyl” refers to an acyl group, as defined herein, attached to an amino group, which is attached to the parent molecular moiety by an alkyl group, as defined herein (i.e., -alkyl-N(R N2 )-C(O)-R, where R is H or optionally substituted C 1-6 , C 1-10 , or C 1-20 is an alkyl group (e.g., haloalkyl), and R N2 is as defined herein). Exemplary unsubstituted acylamino groups contain 1 to 41 carbons (e.g., 1 to 7, 1 to 13, 1 to 21, 2 to 7, 2 to 13, 2 to 21, or 2 to 41 carbons). In some embodiments, the alkyl group is further substituted with 1, 2, 3, or 4 substituents as described herein, and / or the amino group is substituted with -NH or -NHR. N3 and R in the formula N3 are independently OH, NO2, NH2, and NR N4 , SO2OR N4 , SO2R N4 , SOR N4, alkyl, aryl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), or alkoxycarbonylalkyl, and each R N4 can be H, alkyl, or aryl.

[0161] The term "acyloxy" refers to an acyl group, as defined herein, attached to the parent molecular group through an oxygen atom (i.e., -OC(O)-R, where R is H or an optionally substituted C 1-6 , C 1-10 , or C 1-20 (An alkyl group.) Exemplary unsubstituted acyloxy groups contain 1 to 21 carbons (e.g., 1 to 7 or 1 to 11 carbons). In some embodiments, the alkyl group is further substituted with 1, 2, 3, or 4 substituents as described herein.

[0162] The term "acyloxyalkyl" refers to an acyl group, as defined herein, attached to an oxygen atom which is attached to the parent molecular group by an alkyl group (i.e., -alkyl-OC(O)-R, where R is H or an optionally substituted C 1-6 , C 1-10 , or C 1-20 (The acyloxyalkyl group is an alkyl group. Exemplary unsubstituted acyloxyalkyl groups contain 1 to 21 carbons (e.g., 1 to 7 or 1 to 11 carbons). In some embodiments, the alkyl group is further substituted independently with 1, 2, 3, or 4 substituents as described herein.

[0163] The term "alkylaryl" refers to an aryl group, as defined herein, attached to the parent molecular group by an alkyl group, as defined herein. Exemplary unsubstituted alkylaryl groups are those having 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, e.g., C 1-6 Alkyl-C 6-10 Aryl, C 1-10 Alkyl-C 6-10 Aryl, or C 1-20Alkyl-C 6-10 In some embodiments, alkyl and aryl may each be further substituted with 1, 2, 3, or 4 substituents as defined herein for each group. Other groups preceded by the prefix "alkyl" are defined in the same manner, where "alkyl" is defined as C unless otherwise specified. 1-6 The chemical structures referred to and attached to alkyl are as defined herein.

[0164] The term "alkylcycloalkyl" refers to a cycloalkyl group, as defined herein, attached to a parent molecular group by an alkyl group, as defined herein (e.g., an alkyl group having 1 to 4, 1 to 6, 1 to 10, or 1 to 20 carbons). In some embodiments, the alkyl and cycloalkyl can each be further substituted with 1, 2, 3, or 4 substituents, as defined herein, for each group.

[0165] The term "alkenyl" refers to a monovalent straight or branched chain radical containing one or more carbon-carbon double bonds, and having 2 to 20 carbon atoms (e.g., 2 to 6 or 2 to 10 carbons), unless otherwise specified, and is exemplified by ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, and the like. Alkenyl includes both cis and trans isomers. Alkenyl groups can be optionally substituted with 1, 2, 3, or 4 substituents independently selected from amino, aryl, cycloalkyl, or heterocyclyl (e.g., heteroaryl), as defined herein, or any of the exemplary alkyl substituents described herein.

[0166] The term "alkenyloxy" refers to a chemical substituent of formula -OR, where R is C unless otherwise specified. 2-20 Alkenyl groups (e.g., C 2-6 or C 2-10"Alkenyl" refers to a substituent that is an alkyl group (e.g., an alkenyl). Exemplary alkenyloxy groups include ethenyloxy, propenyloxy, and the like. In some embodiments, alkenyl groups can be further substituted with 1, 2, 3, or 4 substituents (e.g., hydroxyl groups) as defined herein.

[0167] "Alkenyl-phosphate" or "alkenyl-phosphorothioate" and the like describe an alkenyl group conjugated to a phosphate group. Those skilled in the art will recognize that such alkenyl groups necessarily contain at least three carbon atoms, and that the alkenyl bond is not adjacent to the phosphate group (i.e., the first and second carbons), thereby resulting in the formation of an enol-phosphate.

[0168] The term "alkylheteroaryl" refers to a heteroaryl group, as defined herein, attached to the parent molecular group by an alkyl group, as defined herein. Exemplary unsubstituted alkylheteroaryl groups include those having 2 to 32 carbons (e.g., 2 to 22, 2 to 18, 2 to 17, 2 to 16, 3 to 15, 2 to 14, 2 to 13, or 2 to 12 carbons, e.g., C 1-6 Alkyl-C 1-12 Heteroaryl, C 1-10 Alkyl-C 1-12 Heteroaryl, or C 1-20 Alkyl-C 1-12 In some embodiments, alkyl and heteroaryl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for each group. Alkylheteroaryl groups are a subset of alkylheterocyclyl groups.

[0169] The term "alkylheterocyclyl" refers to a heterocyclyl group, as defined herein, attached to a parent molecular group by an alkyl group, as defined herein. Exemplary unsubstituted alkylheterocyclyl groups are those having 2 to 32 carbons (e.g., 2 to 22, 2 to 18, 2 to 17, 2 to 16, 3 to 15, 2 to 14, 2 to 13, or 2 to 12 carbons, e.g., C 1-6 Alkyl-C 1-12 Heterocyclyl, C 1-10 Alkyl-C 1-12 Heterocyclyl, or C 1-20 Alkyl-C 1-12 In some embodiments, the alkyl and heterocyclyl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for each group.

[0170] The term "alkoxy" refers to a chemical substituent of formula -OR, where R is C unless otherwise specified. 1-20 Alkyl groups (e.g., C 1-6 or C 1-10 "Alkoxy" refers to a substituent that is an alkyl group. Exemplary alkoxy groups include methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), t-butoxy, and the like. In some embodiments, an alkyl group can be further substituted with 1, 2, 3, or 4 substituents as defined herein (e.g., hydroxy or alkoxy).

[0171] The term "alkoxyalkoxy" refers to an alkoxy group substituted with an alkoxy group. Exemplary unsubstituted alkoxyalkoxy groups are alkoxy groups of 2 to 40 carbons (e.g., 2 to 12 or 2 to 20 carbons, e.g., C 1-6 Alkoxy-C 1-6 Alkoxy, C 1-10 Alkoxy-C 1-10 Alkoxy, or C 1-20 Alkoxy-C 1-20 In some embodiments, each alkoxy group may be further substituted with 1, 2, 3, or 4 substituents as defined herein.

[0172] The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group. Exemplary unsubstituted alkoxyalkyl groups are those having 2 to 40 carbons (e.g., 2 to 12 or 2 to 20 carbons, e.g., C 1-6 Alkoxy-C 1-6 Alkyl, C 1-10 Alkoxy-C 1-10 Alkyl, or C 1-20 Alkoxy-C 1-20 In some embodiments, alkyl and alkoxy can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for each group.

[0173] The term "alkoxycarbonyl" refers to an alkoxy, as defined herein, attached to the parent molecular group through a carbonyl atom (e.g., -C(O)-OR, where R is H or an optionally substituted C 1-6 , C 1-10 , or C 1-20 (Alkoxycarbonyl is an alkyl group. Exemplary unsubstituted alkoxycarbonyls contain 1 to 21 carbons (e.g., 1 to 11 or 1 to 7 carbons). In some embodiments, the alkoxy group is further substituted with 1, 2, 3, or 4 substituents as described herein.

[0174] The term "alkoxycarbonylacyl" refers to an acyl group, as defined herein, substituted with an alkoxycarbonyl group, as defined herein (e.g., -C(O)-alkyl-C(O)-OR, where R is an optionally substituted C 1-6 , C 1-10 , or C 1-20 Exemplary unsubstituted alkoxycarbonyl acyls include 3 to 41 carbons (e.g., 3 to 10, 3 to 13, 3 to 17, 3 to 21, or 3 to 31 carbons, e.g., C 1-6 Alkoxycarbonyl-C 1-6 Achill, C 1-10 Alkoxycarbonyl-C1-10 Acyl, or C 1-20 Alkoxycarbonyl-C 1-20 In some embodiments, the alkoxy and alkyl groups are each further independently substituted with 1, 2, 3, or 4 substituents (e.g., hydroxyl groups) as described herein for each group.

[0175] The term "alkoxycarbonylalkoxy" refers to an alkoxy group, as defined herein, substituted with an alkoxycarbonyl group, as defined herein (e.g., -O-alkyl-C(O)-OR, where R is an optionally substituted C 1-6 , C 1-10 , or C 1-20 Exemplary unsubstituted alkoxycarbonylalkoxy groups include those having 3 to 41 carbons (e.g., 3 to 10, 3 to 13, 3 to 17, 3 to 21, or 3 to 31 carbons, e.g., C 1-6 Alkoxycarbonyl-C 1-6 Alkoxy, C 1-10 Alkoxycarbonyl-C 1-10 Alkoxy, or C 1-20 Alkoxycarbonyl-C 1-20 In some embodiments, each alkoxy group is further independently substituted with 1, 2, 3, or 4 substituents as described herein (e.g., hydroxyl groups).

[0176] The term "alkoxycarbonylalkyl" refers to an alkyl group, as defined herein, substituted with an alkoxycarbonyl group, as defined herein (e.g., -alkyl-C(O)-OR, where R is an optionally substituted C 1-20 , C 1-10 , or C 1-6 Exemplary unsubstituted alkoxycarbonylalkyls are those having 3 to 41 carbons (e.g., 3 to 10, 3 to 13, 3 to 17, 3 to 21, or 3 to 31 carbons, e.g., C 1-6 Alkoxycarbonyl-C 1-6 Alkyl, C 1-10Alkoxycarbonyl-C 1-10 Alkyl, or C 1-20 Alkoxycarbonyl-C 1-20 In some embodiments, the alkyl and alkoxy groups are each further independently substituted with 1, 2, 3, or 4 substituents as described herein (e.g., hydroxyl groups).

[0177] The term "alkoxycarbonylalkenyl" refers to an alkenyl group, as defined herein, substituted with an alkoxycarbonyl group, as defined herein (e.g., -alkenyl-C(O)-OR, where R is an optionally substituted C 1-20 , C 1-10 , or C 1-6 Exemplary unsubstituted alkoxycarbonylalkenyls include those having 4 to 41 carbons (e.g., 4 to 10, 4 to 13, 4 to 17, 4 to 21, or 4 to 31 carbons, e.g., C 1-6 Alkoxycarbonyl-C 2-6 Alkenyl, C 1-10 Alkoxycarbonyl-C 2-10 Alkenyl, or C 1-20 Alkoxycarbonyl-C 2-20 In some embodiments, the alkyl, alkenyl, and alkoxy groups are each further independently substituted with 1, 2, 3, or 4 substituents as described herein (e.g., hydroxyl groups).

[0178] The term "alkoxycarbonylalkynyl" refers to an alkynyl group, as defined herein, substituted with an alkoxycarbonyl group, as defined herein (e.g., -alkynyl-C(O)-OR, where R is an optionally substituted C 1-20 , C 1-10 , or C 1-6 Exemplary unsubstituted alkoxycarbonylalkynyls include those having 4 to 41 carbons (e.g., 4 to 10, 4 to 13, 4 to 17, 4 to 21, or 4 to 31 carbons, e.g., C 1-6 Alkoxycarbonyl-C2-6 Alkynyl, C 1-10 Alkoxycarbonyl-C 2-10 Alkynyl, or C 1-20 Alkoxycarbonyl-C 2-20 In some embodiments, the alkyl, alkynyl, and alkoxy groups are each further independently substituted with 1, 2, 3, or 4 substituents as described herein (e.g., hydroxyl groups).

[0179] The term "alkyl" includes both straight and branched chain saturated groups of 1 to 20 (e.g., 1 to 10 or 1 to 6) carbons unless otherwise specified. Alkyl groups are exemplified by methyl, ethyl, n-propyl and isopropyl, n-, sec-, iso- and tert-butyl, neopentyl, and the like, and optionally include 1, 2, 3, or, in the case of alkyl groups of 2 or more carbons, 4 of the following: (1) C 1-6 Alkoxy; (2) C 1-6 (3) amino, as defined herein (e.g., unsubstituted amino (i.e., —NH) or substituted amino (i.e., —N(R N5 )2, wherein R N6 is as defined for amino); (4) COOaryl-C 1-6 Alkoxy; (5) Azido; (6) Halo; (7) (C 2-9 (8) hydroxy optionally substituted with an O-protecting group; (9) nitro; (10) oxo (e.g., carboxaldehyde or acyl); (11) C 1-7 Spirocyclyl; (12) Thioalkoxy; (13) Thiol; (14) -CO2R optionally substituted with an O-protecting group A’ wherein R A’ is (a)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c) C 6-10 Aryl, (d) hydrogen, (e) C 1-6 Alkyl-C 6-10Aryl, (f) Amino-C 1-20 Alkyl, (g)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 Polyethylene glycol of formula OR', wherein s1 in the formula is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl, and (h)—NR N5 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N5 wherein s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R N5 each independently represents hydrogen or an optionally substituted C 1-6 (15) —C(O)NR B’ RC ’ wherein R B’ and R.C. ’ are each independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 6-10 (16) -SO2R D’ wherein R D’ is (a)C 1-6 Alkyl, (b) C 6-10 Aryl, (c) C 1-6 Alkyl-C 6-10 (17) -SO2NR E’ R F’ wherein R E’ and R F’ are each independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10aryl, and (d) C 1-6 Alkyl-C 6-10 (18) -C(O)R G’ wherein R G’ is (a)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c) C 6-10 Aryl, (d) hydrogen, (e) C 1-6 Alkyl-C 6-10 Aryl, (f) Amino-C 1-20 Alkyl, (g)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 Polyethylene glycol of formula OR', wherein s1 in the formula is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl, and (h)—NR N5 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N5 wherein s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R N5 each independently represents hydrogen or an optionally substituted C 1-6 (19)-NR H’ C(O)R I’ wherein R H’ is (a1) hydrogen and (b1) C 1-6 alkyl; R I’ is (a2)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b2) C 2-20 Alkenyl (e.g., C 2-6alkenyl), (c2) C 6-10 Aryl, (d2) hydrogen, (e2) C 1-6 Alkyl-C 6-10 Aryl, (f2) Amino-C 1-20 Alkyl, (g2)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 Polyethylene glycol of formula OR', wherein s1 in the formula is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl, and (h2)-NR N5 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N5 wherein s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R N5 each independently represents hydrogen or an optionally substituted C 1-6 (20)-NR J’ C(O)OR K’ wherein R J’ is (a1) hydrogen and (b1) C 1-6 alkyl; R K’ is (a2)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b2) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c2) C 6-10 Aryl, (d2) hydrogen, (e2) C 1-6 Alkyl-C 6-10 Aryl, (f2) Amino-C 1-20 Alkyl, (g2)-(CH2) s2 (OCH2CH2) s1 (CH2) s3Polyethylene glycol of formula OR', wherein s1 in the formula is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl, and (h2)-NR N5 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N5 wherein s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R N5 each independently represents hydrogen or an optionally substituted C 1-6 and (21) amidine. In some embodiments, each of these groups may be further substituted as described herein. For example, the alkyl group of a C1-alkylaryl may be further substituted with an oxo group to provide the respective aryl oyl substituent.

[0180] The term "alkylsulfinyl" refers to an alkyl group attached to the parent molecular moiety by an -S(O)- group. Exemplary unsubstituted alkylsulfinyl groups are 1 to 6, 1 to 10, or 1 to 20 carbons. In some embodiments, the alkyl group can be further substituted with 1, 2, 3, or 4 substituents, as defined herein.

[0181] The term "alkylsulfinylalkyl" refers to an alkyl group, as defined herein, substituted with an alkylsulfinyl group. Exemplary unsubstituted alkylsulfinylalkyl groups are 2 to 12, 2 to 20, or 2 to 40 carbons. In some embodiments, each alkyl group can be further substituted with 1, 2, 3, or 4 substituents, as defined herein.

[0182] The term "alkynyl" refers to a monovalent straight or branched chain group of 2 to 20 carbon atoms (e.g., 2 to 4, 2 to 6, or 2 to 10 carbons) containing a carbon-carbon triple bond, and is exemplified by ethynyl, 1-propynyl, etc. Alkynyl groups can be optionally substituted with 1, 2, 3, or 4 substituents independently selected from aryl, cycloalkyl, or heterocyclyl (e.g., heteroaryl), as defined herein, or any of the exemplary alkyl substituents described herein.

[0183] The term "alkynyloxy" refers to a chemical substituent of formula -OR, where R is C unless otherwise specified. 2-20 Alkynyl groups (e.g., C 2-6 or C 2-10 alkynyloxy refers to a substituent that is alkynyloxy (alkynyl). Exemplary alkynyloxy groups include ethynyloxy, propynyloxy, and the like. In some embodiments, alkynyl groups can be further substituted with 1, 2, 3, or 4 substituents as defined herein (e.g., hydroxyl groups).

[0184] The term "amidine" refers to the group -C(=NH)NH2.

[0185] The term "amino" refers to -N(R N6 )2, and R in the formula N6 are independently H, OH, NO2, N(R N7 )2, SO2OR N7 , SO2R N7 , SOR N7, an N-protecting group, alkyl, alkenyl, alkynyl, alkoxy, aryl, alkylaryl, cycloalkyl, alkylcycloalkyl, carboxyalkyl (e.g., optionally substituted with an O-protecting group, e.g., an optionally substituted arylalkoxycarbonyl group or any described herein), sulfoalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), alkoxycarbonylalkyl (e.g., optionally substituted with an O-protecting group, e.g., an optionally substituted arylalkoxycarbonyl group or any described herein), heterocyclyl (e.g., heteroaryl), or alkylheterocyclyl (e.g., alkylheteroaryl), and are substituted with any of the above-listed R N1 Each group may be optionally substituted as defined herein for each group; or two R N1 combine to form a heterocyclyl or N-protecting group, and R N7 are each independently H, alkyl, or aryl. The amino group of the present invention can be unsubstituted amino (i.e., -NH2) or substituted amino (i.e., -N(R')2). In a preferred embodiment, amino is -NH2 or -NHR N5 wherein R N6 are independently OH, NO2, NH2, and NR N7 2. SO2OR N7 , SO2R N7 , SOR N7 , alkyl, carboxyalkyl, sulfoalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), alkoxycarbonylalkyl (e.g., t-butoxycarbonylalkyl), or aryl; R N7 are H and C, respectively. 1-20 Alkyl (e.g., C 1-6 alkyl), or C 1-10 It may be aryl.

[0186] The term "amino acid," as used herein, refers to a molecule having a side chain, an amino group, and an acidic group (e.g., a carboxy group of -COH or a sulfo group of -SOH), where the amino acid is attached to the parent molecular group by the side chain, amino group, or acidic group (e.g., the side chain). In some embodiments, the amino acid is attached to the parent molecular group by a carbonyl group, where the side chain or amino group is attached to the carbonyl group. Exemplary side chains include optionally substituted alkyl, aryl, heterocyclyl, alkylaryl, alkylheterocyclyl, aminoalkyl, carbamoylalkyl, and carboxyalkyl. Exemplary amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, hydroxynorvaline, isoleucine, leucine, lysine, methionine, norvaline, ornithine, phenylalanine, proline, pyrrolysine, selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, and valine. The amino acid group may optionally contain one, two, three, or, in the case of an amino acid group of two or more carbons, four of the following: (1) C 1-6 Alkoxy; (2) C 1-6 (3) amino, as defined herein (e.g., unsubstituted amino (i.e., —NH) or substituted amino (i.e., —N(R N6 )2, where R in the formula N6 is as defined for amino); (4) C 6-10 Aryl-C 1-6 Alkoxy; (5) Azido; (6) Halo; (7) (C 2-9 (heterocyclyl)oxy; (8) hydroxy; (9) nitro; (10) oxo (e.g., carboxaldehyde or acyl); (11) C 1-7 Spirocyclyl; (12) Thioalkoxy; (13) Thiol; (14) -CO2R A’ wherein R A’ is as follows: (a)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b) C 2-20 Alkenyl (e.g., C2-6 alkenyl), (c) C 6-10 Aryl, (d) hydrogen, (e) C 1-6 Alkyl-C 6-10 Aryl, (f) Amino-C 1-20 Alkyl, (g)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 Polyethylene glycol of formula OR', wherein s1 in the formula is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl, and (h)—NR N5 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N5 wherein s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R N5 each independently represents hydrogen or an optionally substituted C 1-6 (15) -C(O)NR B’ RC ’ wherein R B’ and R.C. ’ are each independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 6-10 (16) -SO2R D’ wherein R D’ is (a)C 1-6 Alkyl, (b) C 6-10 Aryl, (c) C 1-6 Alkyl-C 6-10 (17) -SO2NR E’ R F’ wherein R E’and R F’ are each independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 6-10 (18) -C(O)R G’ wherein R G’ is (a)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c) C 6-10 Aryl, (d) hydrogen, (e) C 1-6 Alkyl-C 6-10 Aryl, (f) Amino-C 1-20 Alkyl, (g)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 Polyethylene glycol of formula OR', wherein s1 in the formula is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl, and (h)—NR N5 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N5 wherein s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R N5 each independently represents hydrogen or an optionally substituted C 1-6 alkyl; (19)-NR H’ C(O)R I’ wherein R H’ is (a1) hydrogen and (b1) C 1-6 alkyl; R I’ is (a2)C 1-20 Alkyl (e.g., C1-6 alkyl), (b2) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c2) C 6-10 Aryl, (d2) hydrogen, (e2) C 1-6 Alkyl-C 6-10 Aryl, (f2) Amino-C 1-20 Alkyl, (g2)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 Polyethylene glycol of formula OR', wherein s1 in the formula is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl, and (h2)-NR N5 (CH) s2 (CH2CH2O) s1 (CH2) s3 NR N5 wherein s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R N5 each independently represents hydrogen or an optionally substituted C 1-6 alkyl; (20)-NR J’ C(O)OR K’ wherein R J’ is (a1) hydrogen and (b1) C 1-6 alkyl; R K’ is (a2)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b2) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c2) C 6-10 Aryl, (d2) hydrogen, (e2) C 1-6 Alkyl-C 6-10 Aryl, (f2) Amino-C 1-20 Alkyl, (g2)-(CH2) s2 (OCH2CH2) s1(CH2) s3 Polyethylene glycol of formula OR', wherein s1 in the formula is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl, and (h2)-NR N5 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N5 wherein s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R N5 each independently represents hydrogen or an optionally substituted C 1-6 (21) an alkyl; and (22) an amidine. In some embodiments, each of these groups may be further substituted as described herein.

[0187] The term "aminoalkoxy" refers to an alkoxy group, as defined herein, substituted by an amino group, as defined herein. The alkyl and amino may each be substituted with 1, 2, 3, or 4 substituents, as described herein for the respective group (e.g., COR A’ and R in the formula A’ is (a) C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 Alkyl-C 6-10 It may be further substituted with aryl, for example, selected from the group consisting of carboxy.

[0188] The term "aminoalkyl" refers to an alkyl group, as defined herein, substituted by an amino group, as defined herein. The alkyl and amino may each have one, two, three, or four substituents, as described herein for the respective group (e.g., COR A’ and R in the formula A’ is (a) C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 Alkyl-C 6-10 The aryl may be further substituted with, for example, one selected from the group consisting of carboxy, and / or an N-protecting group.

[0189] The term "aminoalkenyl" refers to an alkenyl group, as defined herein, substituted by an amino group, as defined herein. The alkenyl and amino each may have one, two, three, or four substituents, as described herein for the respective group (e.g., COR A’ and R in the formula A’ is (a) C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 Alkyl-C 6-10 The aryl may be further substituted with, for example, one selected from the group consisting of carboxy, and / or an N-protecting group.

[0190] The term "aminoalkynyl" refers to an alkynyl group, as defined herein, substituted by an amino group, as defined herein. The alkynyl and amino each may have one, two, three, or four substituents, as described herein for the respective group (e.g., COR A’ and R in the formula A’ is (a) C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 Alkyl-C 6-10The aryl may be further substituted with, for example, one selected from the group consisting of carboxy, and / or an N-protecting group.

[0191] The term "aryl" refers to a monocyclic, bicyclic, or polycyclic carbocyclic ring structure having one or two aromatic rings, exemplified by phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, anthracenyl, phenanthrenyl, fluorenyl, indanyl, indenyl, and the like, and optionally containing one, two, three, four, or five substituents, namely: (1) C 1-7 Acyl (e.g., carboxaldehyde); (2) C 1-20 Alkyl (e.g., C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 1-6 Alkylsulfinyl-C 1-6 Alkyl, Amino-C 1-6 Alkyl, azido-C 1-6 Alkyl, (carboxaldehyde)-C 1-6 Alkyl, Halo-C 1-6 Alkyl (e.g., perfluoroalkyl), hydroxy-C 1-6 Alkyl, nitro-C 1-6 Alkyl, or C 1-6 Thioalkoxy-C 1-6 alkyl); (3) C 1-20 Alkoxy (e.g., C such as perfluoroalkoxy) 1-6 Alkoxy); (4) C 1-6 Alkylsulfinyl; (5)C 6-10 Aryl;(6)Amino;(7)C 1-6 Alkyl-C 6-10 Aryl; (8) Azide; (9) C 3-8 Cycloalkyl; (10)C 1-6 Alkyl-C 3-8 Cycloalkyl; (11) halo; (12) C 1-12 Heterocyclyl (e.g., C 1-12 Heteroaryl); (13) (C 1-12(14) Hydroxy; (15) Nitro; (16) C 1-20 Thioalkoxy (e.g., C 1-6 Thioalkoxy);(17)-(CH2) q CO2R A’ In the formula, q is an integer of 0 to 4, and R A’ is (a)C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 Alkyl-C 1-10 (18)-(CH2) q CONR B’ R C’ In the formula, q is an integer of 0 to 4, and R B’ and R C’ are independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 6-10 (19)-(CH2) q SO2R D’ In the formula, q is an integer of 0 to 4, and R D’ is (a) alkyl, (b) C 6-10 aryl, and (c) alkyl-C 6-10 (20)-(CH2) q SO2NR E’ R F’ wherein q is an integer from 0 to 4, and R E’ and R F’ are each independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 6-10 (21) thiol; (22) C 6-10 Aryloxy; (23)C 3-8 Cycloalkoxy; (24)C 6-10 Aryl-C 1-6 Alkoxy; (25)C 1-6 Alkyl-C 1-12Heterocyclyl (e.g., C 1-6 Alkyl-C 1-12 Heteroaryl);(26)C 2-20 Alkenyl; and (27)C 1-20 and alkynyl. In some embodiments, each of these groups may be further substituted as described herein. For example, the alkyl group of a C1-alkylaryl or C1-alkylheterocyclyl can be further substituted with an oxo group to give the respective aryl and (heterocyclyl)oyl substituents.

[0192] The term "arylalkoxy" refers to an alkylaryl group, as defined herein, attached to the parent molecular group through an oxygen atom. Exemplary unsubstituted arylalkoxy groups are alkylaryl groups having 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, e.g., C 6-10 Aryl-C 1-6 Alkoxy, C 6-10 Aryl-C 1-10 Alkoxy, or C 6-10 Aryl-C 1-20 In some embodiments, arylalkoxy groups can be substituted with 1, 2, 3, or 4 substituents as defined herein.

[0193] The term "arylalkoxycarbonyl" refers to an arylalkoxy group, as defined herein, attached to the parent molecular group through a carbonyl (e.g., -C(O)-O-alkyl-aryl). Exemplary unsubstituted arylalkoxy groups are those having 8 to 31 carbons (e.g., 8 to 17 or 8 to 21 carbons, e.g., C 6-10 Aryl-C 1-6 Alkoxy-carbonyl, C 6-10 Aryl-C 1-10 Alkoxy-carbonyl, or C 6-10 Aryl-C 1-20In some embodiments, arylalkoxycarbonyl groups can be substituted with 1, 2, 3, or 4 substituents, as defined herein.

[0194] The term "aryloxy" refers to a chemical substituent of formula -OR', where R' is an aryl group containing 6 to 18 carbon atoms, unless otherwise specified. In some embodiments, the aryl group can be substituted with 1, 2, 3, or 4 substituents as defined herein.

[0195] The term "aryloyl" refers to an aryl group, as defined herein, attached to the parent molecular moiety by a carbonyl group. Exemplary unsubstituted aryloyl groups are those containing 7 to 11 carbons. In some embodiments, the aryl group can be substituted with 1, 2, 3, or 4 substituents, as defined herein.

[0196] The term "azido" refers to the group -N3, which can also be represented as -N=N=N.

[0197] The term "bicyclic" refers to a structure having two rings, which may or may not be aromatic. A bicyclic structure comprises a spirocyclyl group, as defined herein, and two rings sharing one or more bridges, which may comprise one atom or a chain containing two, three, or more atoms. Exemplary bicyclic groups include bicyclic carbocyclyl groups, as defined herein, in which the first and second rings are carbocyclyl groups; bicyclic aryl groups, as defined herein, in which the first and second rings are aryl groups; bicyclic heterocyclyl groups, in which the first ring is a heterocyclyl group and the second ring is a carbocyclyl (e.g., aryl) or heterocyclyl (e.g., heteroaryl) group; and bicyclic heteroaryl groups, in which the first ring is a heteroaryl group and the second ring is a carbocyclyl (e.g., aryl) or heterocyclyl (e.g., heteroaryl) group. In some embodiments, bicyclic groups can be substituted with 1, 2, 3, or 4 substituents as defined herein for cycloalkyl, heterocyclyl, and aryl groups.

[0198] The term "boranyl" refers to -B(R B1 )3, and R B1 are each independently selected from the group consisting of H and optionally substituted alkyl. In some embodiments, the boranyl group can be substituted with 1, 2, 3, or 4 substituents as defined herein for alkyl groups.

[0199] The terms "carbocyclic" and "carbocyclyl" refer to an optionally substituted C 3-12 refers to monocyclic, bicyclic, or tricyclic structures in which the rings are formed by carbon atoms, which may or may not be aromatic. Carbocyclic structures include cycloalkyl, cycloalkenyl, and aryl groups.

[0200] The term "carbamoyl" refers to -C(O)-N(R N1 )2, where each R N1The meaning of is found in the definition of "amino" provided herein.

[0201] The term "carbamoylalkyl" refers to an alkyl group, as defined herein, substituted by a carbamoyl group, as defined herein. The alkyl group may be further substituted with one, two, three, or four substituents, as described herein.

[0202] The term "carbamyl" has the structure -NR N8 C(=O)OR or -OC(=O)N(R N8 ) 2, each R N8 is found in the definition of "amino" provided herein, and R is alkyl, cycloalkyl, alkylcycloalkyl, aryl, alkylaryl, heterocyclyl (e.g., heteroaryl), or alkylheterocyclyl (e.g., alkylheteroaryl), as defined herein.

[0203] The term "carbonyl" refers to the group C(O), which can also be represented as C=O.

[0204] The term "carboxaldehyde" refers to an acyl group having the structure -CHO.

[0205] The term "carboxy" refers to -CO2H.

[0206] The term "carboxyalkoxy" refers to an alkoxy group, as defined herein, substituted by a carboxy group, as defined herein. The alkoxy group can be further substituted with one, two, three, or four substituents, as described herein for alkyl groups, and the carboxy group can optionally be substituted with one or more O-protecting groups.

[0207] The term "carboxyalkyl" refers to an alkyl group, as defined herein, substituted by a carboxy group, as defined herein. The alkyl group can be further substituted with one, two, three, or four substituents, as described herein, and the carboxy group can optionally be substituted with one or more O-protecting groups.

[0208] The term "carboxyaminoalkyl" refers to an aminoalkyl group, as defined herein, substituted by carboxy, as defined herein. Carboxy, alkyl, and amino each may have one, two, three, or four substituents, as described herein for the respective group (e.g., COR A’ and R in the formula A’ is (a) C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 Alkyl-C 6-10 aryl, for example, selected from the group consisting of carboxy, an N-protecting group, and / or an O-protecting group.

[0209] The term "cyano" refers to the group --CN.

[0210] The term "cycloalkoxy" refers to a chemical substituent of formula -OR, where R is a C, as defined herein unless otherwise specified. 3-8 "Cycloalkoxy" refers to a substituent that is a cycloalkyl group. The cycloalkyl group may be further substituted with one, two, three, or four substituents as described herein. Exemplary unsubstituted cycloalkoxy groups contain 3 to 8 carbon atoms. In some embodiments, the cycloalkyl group may be further substituted with one, two, three, or four substituents as described herein.

[0211] The term "cycloalkyl," unless otherwise specified, refers to a monovalent saturated or unsaturated non-aromatic cyclic hydrocarbon group of 3 to 8 carbon atoms, and is exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclic heptyl, and the like. When the cycloalkyl group contains one carbon-carbon double bond, the cycloalkyl group can be referred to as a "cycloalkenyl" group. Exemplary cycloalkenyl groups include cyclopentenyl, cyclohexenyl, and the like. Cycloalkyl groups of the present invention optionally include: (1) C 1-7 Acyl (e.g., carboxaldehyde); (2) C 1-20 Alkyl (e.g., C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 1-6 Alkylsulfinyl-C 1-6 Alkyl, Amino-C 1-6 Alkyl, azido-C 1-6 Alkyl, (carboxaldehyde)-C 1-6 Alkyl, Halo-C 1-6 Alkyl (e.g., perfluoroalkyl), hydroxy-C 1-6 Alkyl, nitro-C 1-6 Alkyl, or C 1-6 Thioalkoxy-C 1-6 alkyl); (3) C 1-20 Alkoxy (e.g., C such as perfluoroalkoxy) 1-6 Alkoxy); (4) C 1-6 Alkylsulfinyl; (5)C 6-10 Aryl;(6)Amino;(7)C 1-6 Alkyl-C 6-10 Aryl; (8) Azide; (9) C 3-8 Cycloalkyl; (10)C 1-6 Alkyl-C 3-8 Cycloalkyl; (11) halo; (12) C 1-12 Heterocyclyl (e.g., C 1-12 Heteroaryl); (13) (C 1-12 (14) Hydroxy; (15) Nitro; (16) C 1-20 Thioalkoxy (e.g., C 1-6Thioalkoxy);(17)-(CH2) q CO2R A’ In the formula, q is an integer of 0 to 4, and R A’ is (a)C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 Alkyl-C 6-10 (18)-(CH2) q CONR B’ R C’ In the formula, q is an integer of 0 to 4, and R B’ and R C’ are independently (a) hydrogen, (b) C 6-10 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 6-10 (19)-(CH2) q SO2R D’ In the formula, q is an integer of 0 to 4, and R D’ is (a)C 6-10 Alkyl, (b) C 6-10 aryl, and (c) C 1-6 Alkyl-C 6-10 (20)-(CH2) q SO2NR E’ R F’ In the formula, q is an integer of 0 to 4, and R E’ and R F’ are each independently (a) hydrogen, (b) C 6-10 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 1-10 (21) thiol; (22) C 6-10 Aryloxy; (23)C 3-8 Cycloalkoxy; (24)C 6-10 Aryl-C 1-6 Alkoxy; (25)C 1-6 Alkyl-C 1-12 Heterocyclyl (e.g., C 1-6 Alkyl-C 1-12Heteroaryl; (26) oxo; (27) C 2-20 Alkenyl; and (28)C 2-20 In some embodiments, each of these groups may be further substituted as described herein. For example, a C1-alkylaryl or C1-alkylheterocyclyl alkyl group may be further substituted with an oxo group to give the respective aryloyl and (heterocyclyl)oyl substituents.

[0212] The term "diastereomers" refers to stereoisomers that are not mirror images of each other and are not superimposable with respect to one another.

[0213] The term "effective amount" of an agent is an amount sufficient to achieve a beneficial or desired result, e.g., a clinical result, and therefore, "effective amount" varies depending on the context in which the term is applied. For example, in the context of administering an agent to treat cancer, an effective amount of the agent is an amount sufficient to achieve treatment, as defined herein, of the cancer, e.g., compared to the response obtained without administration of the agent.

[0214] The term "enantiomer" refers to each individual optically active form of a compound of the present disclosure having an optical purity or enantiomeric excess (as determined by standard methods in the art) of at least 80% (i.e., at least 90% of one enantiomer and at most 10% of the other enantiomer), preferably at least 90%, and more preferably at least 98%.

[0215] The term "halo" refers to a halogen selected from bromine, chlorine, iodine, or fluorine.

[0216] The term "haloalkoxy" refers to an alkoxy group, as defined herein, substituted with a halogen group (i.e., F, Cl, Br, or I). The haloalkoxy can be substituted with one, two, three, or, in the case of alkyl groups of two or more carbon atoms, four halogens. Haloalkoxy groups include perfluoroalkoxy (e.g., -OCF), -OCHF, -OCHF, -OCCl, -OCHCHBr, -OCHCH(CHCHBr)CH, and -OCHICH. In some embodiments, the haloalkoxy group can be further substituted with one, two, three, or four substituents, as described herein for alkyl groups.

[0217] The term "haloalkyl" refers to an alkyl group, as defined herein, substituted with a halogen group (i.e., F, Cl, Br, or I). Haloalkyl can be substituted with one, two, three, or, in the case of alkyl groups of two or more carbon atoms, four halogens. Haloalkyl groups include perfluoroalkyl (e.g., -CF), -CHF, -CHF, -CCl, -CHCHBr, -CHCH(CHCHBr)CH, and -CHICH. In some embodiments, haloalkyl groups can be further substituted with one, two, three, or four substituents, as described herein for alkyl groups.

[0218] The term "heteroalkyl" refers to an alkyl group, as defined herein, in which one or two of the constituent carbon atoms are replaced by nitrogen, oxygen, or sulfur, respectively. In some embodiments, heteroalkyl groups can be further substituted with one, two, three, or four substituents, as described herein for alkyl groups.

[0219] The term "heteroaryl" refers to a subset of heterocyclyl, as defined herein, that is aromatic, i.e., contains 4n+2 π-electrons in a monocyclic or polycyclic ring structure. Exemplary unsubstituted heteroaryl groups contain 1 to 12 carbon atoms (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9). In some embodiments, heteroaryl is substituted with 1, 2, 3, or 4 substituents as defined for heterocyclyl groups.

[0220] The term "heterocyclyl," unless otherwise specified, refers to a 5-, 6-, or 7-membered ring containing 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Five-membered rings have 0 to 2 double bonds, and 6- and 7-membered rings have 0 to 3 double bonds. Exemplary unsubstituted heterocyclyl groups have 1 to 12 carbon atoms (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9). The term "heterocyclyl" also refers to bridged polycyclic structures in which one or more carbon and / or heteroatoms bridge two non-adjacent members of a ring (e.g., a quinuclidinyl group). The term "heterocyclyl" includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocycles is fused to one, two, or three carbon rings, such as an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, or another monocyclic heterocycle, such as indolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, benzofuryl, benzothienyl, etc. Examples of fused heterocyclyls include tropanes and 1,2,3,5,8,8a-hexahydroindolizine. Heterocycles include pyrrolyl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, piperidinyl, homopiperidinyl, pyrazinyl, piperazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiomorpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl nyl, indolyl, indazolyl, quinolyl, isoquinolyl, quinoxalinyl, dihydroquinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, benzothiadiazolyl, furyl, thienyl, thiazolidinyl, isothiazolyl, triazolyl, tetrazolyl, oxadiazolyl (e.g., 1,2,3-oxadiazolyl), purinyl, thiadiazolyl (e.g., 1,2,3-thiadiazolyl), tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, dihydroindolyl, dihydroquinolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, dihydroisoquinolyl, pyranyl, dihydropyranyl, dithiazolyl, benzofuranyl, isobenzofuranyl, benzothienyl, and the like, including dihydro and tetrahydro forms thereof where one or more double bonds are reduced and replaced with hydrogen. Further exemplary heterocyclyls include 2,3,4,5-tetrahydro-2-oxo-oxazolyl; 2,3-dihydro-2-oxo-1H-imidazolyl; 2,3,4,5-tetrahydro-5-oxo-1H-pyrazolyl (e.g., 2,3,4,5-tetrahydro-2-phenyl-5-oxo-1H-pyrazolyl); 2,3,4,5-tetrahydro-2,4-dioxo-1H-imidazolyl (e.g., 2,3,4,5-tetrahydro-2-phenyl-5-oxo-1H-pyrazolyl); Hydro-2,4-dioxo-5-methyl-5-phenyl-1H-imidazolyl; 2,3-dihydro-2-thioxo-1,3,4-oxadiazolyl (e.g., 2,3-dihydro-2-thioxo-5-phenyl-1,3,4-oxadiazolyl); 4,5-dihydro-5-oxo-1H-triazolyl (e.g., 4,5-dihydro-3-methyl-4-amino-5-oxo-1H-triazolyl); 1,2,3,4-tetrahydro-2, 4-Dioxopyridinyl (e.g., 1,2,3,4-tetrahydro-2,4-dioxo-3,3-diethylpyridinyl); 2,6-dioxo-piperidinyl (e.g., 2,6-dioxo-3-ethyl-3-phenylpiperidinyl); 1,6-dihydro-6-oxopyridinyl; 1,6-dihydro-4-oxopyrimidinyl (e.g., 2-(methylthio)-1,6-dihydro-4-oxo-5-methylpyrimidin-1-yl); 1, 2,3,4-tetrahydro-2,4-dioxopyrimidinyl (e.g., 1,2,3,4-tetrahydro-2,4-dioxo-3-ethylpyrimidinyl); 1,6-dihydro-6-oxo-pyridazinyl (e.g., 1,6-dihydro-6-oxo-3-ethylpyridazinyl); 1,6-dihydro-6-oxo-1,2,4-triazinyl (e.g., 1,6-dihydro-5-isopropyl-6-oxo-1,2,4-triazinyl); 2,3-Dihydro-2-oxo-1H-indolyl (e.g., 3,3-dimethyl-2,3-dihydro-2-oxo-1H-indolyl and 2,3-dihydro-2-oxo-3,3'-spiropropane-1H-indol-1-yl); 1,3-dihydro-1-oxo-2H-iso-indolyl; 1,3-dihydro-1,3-dioxo-2H-iso-indolyl; 1H-benzopyrazolyl (e.g., 1-(ethoxycarbonyl)-1H-benzopyrazolyl) );2,3-dihydro-2-oxo-1H-benzimidazolyl (e.g., 3-ethyl-2,3-dihydro-2-oxo-1H-benzimidazolyl);2,3-dihydro-2-oxo-benzoxazolyl (e.g., 5-chloro-2,3-dihydro-2-oxo-benzoxazolyl);2,3-dihydro-2-oxo-benzoxazolyl;2-oxo-2H-benzopyranyl;1,4-benzodioxanyl;1,3-benzodioxanyl;2 ,3-Dihydro-3-oxo,4H-1,3-benzothiazinyl; 3,4-Dihydro-4-oxo-3H-quinazolinyl (e.g., 2-methyl-3,4-dihydro-4-oxo-3H-quinazolinyl); 1,2,3,4-tetrahydro-2,4-dioxo-3H-quinazolyl (e.g., 1-ethyl-1,2,3,4-tetrahydro-2,4-dioxo-3H-quinazolyl); 1,2,3,6-tetrahydro-2,6-dioxo-7H-purinyl (e.g., For example, 1,2,3,6-tetrahydro-1,3-dimethyl-2,6-dioxo-7H-purinyl; 1,2,3,6-tetrahydro-2,6-dioxo-1H-purinyl (e.g., 1,2,3,6-tetrahydro-3,7-dimethyl-2,6-dioxo-1H-purinyl); 2-oxobenz[c,d]indolyl; 1,1-dioxo-2H-naphtho[1,8-c,d]isothiazolyl; and 1,8-naphthylenedicarboxamide. Further heterocycles include 3,3a,4,5,6,6a-hexahydro-pyrrolo[3,4-b]pyrrol-(2H)-yl, and 2,5-diazabicyclo[2.2.1]heptano-2-yl, homopiperazinyl (or diazepanyl), tetrahydropyranyl, dithiazolyl, benzofuranyl, benzothienyl, oxepanyl, thiepanyl, azocanyl, oxecanyl, and thiocanyl. Heterocyclic groups further include those of the formula: [ka]

[0221] wherein E' is selected from the group consisting of -N- and -CH-; F' is selected from the group consisting of -N=CH-, -NH-CH2-, -NH-C(O)-, -NH-, -CH=N-, -CH2-NH-, -C(O)-NH-, -CH=CH-, -CH2-, -CH2CH2-, -CHO-, -OCH2-, -O-, and -S-; and G' also includes a group selected from the group consisting of -CH- and -N-.

[0222] Any heterocyclyl group referred to herein may optionally contain 1, 2, 3, 4 or 5 of the following: (1) C 1-7 Acyl (e.g., carboxaldehyde); (2) C 1-20 Alkyl (e.g., C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 1-6 Alkylsulfinyl-C 1-6 Alkyl, Amino-C 1-6 Alkyl, azido-C 1-6 Alkyl, (carboxaldehyde)-C 1-6 Alkyl, Halo-C 1-6 Alkyl (e.g., perfluoroalkyl), hydroxy-C 1-6 Alkyl, nitro-C 1-6 Alkyl, or C 1-6 Thioalkoxy-C 1-6 alkyl); (3) C 1-20 Alkoxy (e.g., C such as perfluoroalkoxy) 1-6 Alkoxy); (4) C 1-6 Alkylsulfinyl; (5)C 6-10 Aryl;(6)Amino;(7)C 1-6 Alkyl-C 6-10 Aryl; (8) Azide; (9) C 3-8 Cycloalkyl; (10)C 1-6 Alkyl-C 3-8 Cycloalkyl; (11) halo; (12) C1-12 Heterocyclyl (e.g., C 2-12 Heteroaryl); (13) (C 1-12 (14) Hydroxy; (15) Nitro; (16) C 1-20 Thioalkoxy (e.g., C 1-6 Thioalkoxy);(17)-(CH2) q CO2R A’ In the formula, q is an integer of 0 to 4, and R A’ is (a)C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 Alkyl-C 6-10 (18)-(CH2) q CONR B’ R C’ In the formula, q is an integer of 0 to 4, and R B’ and R C’ are independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 6-10 (19)-(CH2) q SO2R D’ In the formula, q is an integer of 0 to 4, and R D’ is (a)C 1-6 Alkyl, (b) C 6-10 aryl, and (c) C 1-6 Alkyl-C 6-10 (20)-(CH2) q SO2NR E’ R F’ In the formula, q is an integer of 0 to 4, and R E’ and R F’ are each independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 Alkyl-C 6-10 (21) thiol; (22) C 6-10 Aryloxy; (23)C3-8 Cycloalkoxy; (24) Arylalkoxy; (25) C 1-6 Alkyl-C 1-12 Heterocyclyl (e.g., C 1-6 Alkyl-C 1-12 Heteroaryl; (26) oxo; (27) (C 1-12 Heterocyclyl)imino; (28)C 2-20 Alkenyl; and (29)C 2-20 and alkynyl. In some embodiments, each of these groups may be further substituted as described herein. For example, the alkyl group of a C1-alkylaryl or C1-alkylheterocyclyl may be further substituted with an oxo group to give the respective aryl and (heterocyclyl)oyl substituents.

[0223] The term "(heterocyclyl)imino" refers to a heterocyclyl group, as defined herein, attached to a parent molecular group by an imino group. In some embodiments, the heterocyclyl group can be further substituted with 1, 2, 3, or 4 substituents, as defined herein.

[0224] The term "(heterocyclyl)oxy" refers to a heterocyclyl group, as defined herein, attached to the parent molecular group by an oxygen atom. In some embodiments, the heterocyclyl group can be further substituted with 1, 2, 3, or 4 substituents, as defined herein.

[0225] The term "(heterocyclyl)oyl" refers to a heterocyclyl group, as defined herein, attached to a parent molecular group by a carbonyl group. In some embodiments, the heterocyclyl group can be further substituted with 1, 2, 3, or 4 substituents, as defined herein.

[0226] The term "hydrocarbon" refers to a group consisting solely of carbon and hydrogen atoms.

[0227] The term "hydroxy" refers to an -OH group. In some embodiments, a hydroxy group (hydroxyl group) can be substituted with 1, 2, 3, or 4 substituents as defined herein for alkyl (e.g., O-protecting groups).

[0228] The term "hydroxyalkenyl" refers to an alkenyl group, as defined herein, substituted with one to three hydroxyl groups, provided that no more than one hydroxyl group may be attached to a carbon atom of the alkyl group, and is exemplified by dihydroxypropenyl, hydroxyisopentenyl, etc. In some embodiments, the hydroxyalkenyl group may be substituted with one, two, three, or four substituents, as defined herein for alkyl (e.g., O-protecting groups).

[0229] The term "hydroxyalkyl" refers to an alkyl group, as defined herein, substituted with one to three hydroxyl groups, provided that no more than one hydroxyl group may be attached to a carbon atom of the alkyl group, and is exemplified by hydroxymethyl, dihydroxypropyl, etc. In some embodiments, the hydroxyalkyl group may be substituted with one, two, three, or four substituents as defined herein for alkyl (e.g., O-protecting groups).

[0230] The term "hydroxyalkynyl" refers to an alkynyl group, as defined herein, substituted with one to three hydroxyl groups, provided that no more than one hydroxyl group may be attached to a carbon atom of the alkyl group. In some embodiments, the hydroxyalkynyl group may be substituted with one, two, three, or four substituents, as defined herein for alkyl (e.g., O-protecting groups).

[0231] The term "isomer" refers to any tautomer, stereoisomer, enantiomer, or diastereomer of any compound of the present disclosure. It is recognized that compounds of the present disclosure can have one or more chiral centers and / or double bonds and thus can exist as stereoisomers, e.g., double bond isomers (i.e., geometric E / Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers). In accordance with the present disclosure, the chemical structures depicted herein and, therefore, the compounds of the present disclosure encompass all of the corresponding stereoisomers, i.e., stereomerically pure forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure), as well as mixtures of enantiomers and stereoisomers (e.g., racemates). Enantiomeric and stereoisomeric mixtures of the compounds of the present disclosure can typically be resolved into the component enantiomers or stereoisomers of the mixture by well-known methods, such as chiral gas chromatography, chiral high performance liquid chromatography, crystallization of the compound as a chiral salt complex, or crystallization of the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained from stereomerically pure or enantiomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthetic methods.

[0232] The term "monosaccharide" refers to a simple sugar. Examples of monosaccharides include allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, D-fucitol, L-fucitol, fucosamine, fucose, fuculose, galactosamine, D-galactosaminitol, N-acetyl-galactosamine, galactose, glucosamine, N-acetyl-glucosamine, glucosaminitol, glucose, glucose-6-phosphate ... Monosaccharides include, but are not limited to, D-glycero-D-mannose-heptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, mannose, mannose-6-phosphate, psicose, quinovose, quinovosamine, rhamnitol, rhamnosamine, rhamnose, ribose, ribulose, sedoheptulose, sorbose, tagatose, talose, tartaric acid, threose, xylose, and xylulose. Monosaccharides can be in the D or L configuration. The term "monosaccharide derivative" refers to deoxysugars (where an alcohol hydroxy group is replaced by hydrogen), aminosugars (where an alcohol hydroxy group is replaced by an amino group), thiosugars (where an alcohol hydroxy group is replaced by a thiol, or where C=O is replaced by C=S, or where a ring oxygen in a cyclic form is replaced by sulfur), selenosugars, tellurosugars, azasugars (where a ring carbon is replaced by nitrogen), aminosugars (where a ring oxygen is replaced by nitrogen), phosphanosugars (where a ring oxygen is replaced by phosphorus), phosphasugars (where a ring carbon is replaced by phosphorus), C-substituted monosaccharides (where a hydrogen at a non-terminal carbon atom is replaced by carbon), unsaturated monosaccharides, alditols (where a carbonyl group is replaced by a CHOH group), aldonic acids (where an aldehyde group is replaced by a carboxy group), ketoaldonic acids, uronic acids, aldaric acids, and the like. Amino sugars include amino monosaccharides, preferably galactosamine, glucosamine, mannosamine, fucosamine, quinovosamine, neuraminic acid, muramic acid, lactosediamine, acosamine, bacillosamine, daunosamine, desosamine, forosamine, galosamine, kanosamine, kansosamine, mycaminose, mycosamine, perosamine, pneumosamine, purpurosamine, and rhodosamine. It is understood that such monosaccharides can be further substituted.

[0233] The term "disaccharide" refers to a dimer consisting of two monosaccharide residues joined by a carbohydrate bond. Examples of disaccharides include, but are not limited to, sucrose, lactose, isomaltulose, maltose, trehalose, and trehalulose.

[0234] The term "vitamin" refers to an organic molecule that is an essential micronutrient. Examples of vitamins include retinol, retinal, retinyl esters, and retinoic acid; vitamin B3 and nicotinic acid; biotin; vitamin B2; vitamin B3 and nicotinic acid; vitamin B5; vitamin B6, pyridoxal, and pyridoxine; vitamin B 12 Vitamin C; choline; vitamin D; vitamin E; vitamin B9, formic acid and folic acid; and vitamin K and glutathione.

[0235] The term "polyol" refers to sugar alcohols such as, but not limited to, maltitol, sorbitol, mannitol, lactitol, xylitol, erythritol, and isomalt; and monomeric polyols such as, but not limited to, glycerin, pentaerythritol, ethylene glycol, and sucrose.

[0236] The term "polysialic acid" refers to a naturally occurring, unbranched polymer of sialic acid produced by certain bacterial strains and in certain mammalian cells. The abbreviation "PSA" used herein refers to the term "polysialic acid." Similarly, the term "mPSA" used herein refers to the term "modified polysialic acid" or a derivative of polysialic acid. PSA consists of a polymer (usually a homopolymer) of N-acetylneuraminic acid. The secondary amino group usually bears an acetyl group, but may alternatively bear a glycolyl group in mPSA. Possible substitutions for the hydroxyl group to form mPSA include acetyl, lactyl, ethyl, sulfate, and phosphate groups. [ka]

[0237] PSA and mPSA generally comprise linear polymers consisting essentially of N-acetylneuraminic acid moieties linked by 2,8- or 2,9-glycosidic linkages, or combinations thereof (e.g., alternating 2,8- and 2,9-glycosidic linkages). In particularly preferred PSA and mPSA, the glycosidic linkages are α-2,8. Such PSAs and mPSAs are conveniently derived from colominic acid and are referred to herein as "CA" and "mCA." Typical PSAs and mPSAs contain at least 2, preferably at least 5, more preferably at least 10, and most preferably at least 20 N-acetylneuraminic acid moieties. Thus, PSAs and mPSAs may contain from 5 to 500 N-acetylneuraminic acid moieties, preferably from 10 to 300. PSAs and CAs may be polymers containing different sugar moieties. PSAs and CAs may also be copolymers. The PSA and CA are preferably essentially free of sugar moieties other than N-acetylneuraminic acid. The PSA and CA preferably contain at least 90%, more preferably at least 95%, and most preferably at least 98% N-acetylneuraminic acid moieties.

[0238] When PSAs and CAs contain moieties other than N-acetylneuraminic acid (e.g., as in mPSAs and mCAs), these moieties are preferably located at one or both ends of the polymer chain. For example, such "other" moieties can be moieties derived from the terminal N-acetylneuraminic acid moiety by oxidation or reduction.

[0239] For example, WO 2001 / 087922 describes mPSA and mCA in which the non-reducing terminal N-acetylneuraminic acid unit is converted to an aldehyde group by reaction with sodium periodate. Furthermore, WO 2005 / 016974 describes mPSA and mCA in which the reducing terminal N-acetylneuraminic acid unit is reduced, reductively opening the reducing terminal N-acetylneuraminic acid unit, thereby forming a vicinal diol group, which is subsequently oxidized to convert the vicinal diol group to an aldehyde group.

[0009] Sialic acid-rich glycoproteins bind selectins in humans and other organisms. They play an important role in human influenza infection. For example, sialic acid can hide mannose antigens on the surface of host cells or bacteria from mannose-binding lectins. This prevents complement activation. Sialic acid also masks the galactose residue in the penultimate position, preventing rapid clearance of the glycoprotein by the galactose receptor on hepatocytes. [ka]

[0240] The term "derivative" refers to, but is not limited to, any compound having a structure derived from a compound of the present disclosure, which structure is sufficiently similar to that disclosed herein and, based on that similarity, would be expected by one of ordinary skill in the art to exhibit the same or similar activity and utility as the claimed and / or referenced compound.

[0241] The term "N-protected amino" refers to an amino group, as defined herein, having one or two N-protecting groups, as defined herein, attached thereto.

[0242] The term "N-protecting group" refers to a group intended to protect an amino group against undesired reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, "Protective Groups in Organic Synthesis", 3rd Edition (John Wiley & Sons, New York, 1999), 1999, which is incorporated herein by reference.N-protecting groups include acyl, aryloyl, or carbamyl groups, such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and chiral auxiliaries, such as protected or unprotected D,L or D,L amino acids, such as , alanine, leucine, phenylalanine, etc.; sulfonyl-containing groups, for example, benzenesulfonyl, p-toluenesulfonyl, etc.; carbamate-forming groups, for example, benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-Dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl , methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl, etc.; alkylaryl groups such as benzyl, triphenylmethyl, benzyloxymethyl, etc.; and silyl groups such as trimethylsilyl, etc.Preferred N-protecting groups are formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

[0243] The term "nitro" refers to the group --NO.sub.2.

[0244] The term "O-protecting group" refers to a group intended to protect an oxygen-containing (e.g., phenol, hydroxyl, or carbonyl) group from undesired reactions during synthetic procedures. Commonly used O-protecting groups are disclosed in Greene and Wuts, "The Role of Protective Groups in Organic Synthesis," 5th Edition (John Wiley & Sons, New York, 2014), which is incorporated herein by reference. Exemplary O-protecting groups include acyl, aryloyl, or carbamyl groups, such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, t-butyldimethylsilyl, tri-isopropylsilyloxymethyl, 4,4′-dimethoxytrityl, isobutyryl, phenoxyacetyl, 4-isopropylphenoxyacetyl, dimethylformamidino, and 4-nitrobenzoyl; alkylcarbonyl groups, such as acyl, acetyl, propionyl, pivaloyl, and the like; and optionally substituted arylcarbonyl groups, such as , benzoyl; silyl groups, such as trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-isopropylsilyloxymethyl (TOM), triisopropylsilyl (TIPS), etc.; groups that form ethers with hydroxyl, such as methyl, methoxymethyl, tetrahydropyranyl, benzyl, p-methoxybenzyl, trityl, etc.; alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, n-isopropoxycarbonyl, n-butyloxycarbonyl, isobutyloxycarbonyl, sec-butyloxycarbonyl, t-butyloxycarbonyl, 2-ethylhexyloxycarbonyl, cyclohexyloxycarbonyl, methyloxycarbonyl, etc.;Alkoxyalkoxycarbonyl groups, such as methoxymethoxycarbonyl, ethoxymethoxycarbonyl, 2-methoxyethoxycarbonyl, 2-ethoxyethoxycarbonyl, 2-butoxyethoxycarbonyl, 2-methoxyethoxymethoxycarbonyl, allyloxycarbonyl, propargyloxycarbonyl, 2-buteneoxycarbonyl, 3-methyl-2-buteneoxycarbonyl, etc.; haloalkoxycarbonyl groups, such as 2-chloroethoxycarbonyl, 2-chloroethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, etc. optionally substituted arylalkoxycarbonyl groups, such as benzyloxycarbonyl, p-methylbenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2,4-dinitrobenzyloxycarbonyl, 3,5-dimethylbenzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, fluorenylmethyloxycarbonyl, and the like; and optionally substituted aryloxycarbonyl groups, such as phenoxycarbonyl, p-nitrophenoxycarbonyl, dicarbonyl, o-nitrophenoxycarbonyl, 2,4-dinitrophenoxycarbonyl, p-methylphenoxycarbonyl, m-methylphenoxycarbonyl, o-bromophenoxycarbonyl, 3,5-dimethylphenoxycarbonyl, p-chlorophenoxycarbonyl, 2-chloro-4-nitrophenoxycarbonyl, etc.; substituted alkyl, aryl, and alkylaryl ethers (e.g., trityl; methylthiomethyl; methoxymethyl; benzyloxymethyl; siloxymethyl; 2,2,2-trichloroethoxymethyl; tetra ... hydropyranyl; tetrahydrofuranyl; ethoxyethyl; 1-[2-(trimethylsilyl)ethoxy]ethyl; 2-trimethylsilylethyl; t-butyl ether; p-chlorophenyl, p-methoxyphenyl, p-nitrophenyl, benzyl, p-methoxybenzyl, and nitrobenzyl; silyl ethers (e.g., trimethylsilyl; triethylsilyl; triisopropylsilyl; dimethylisopropylsilyl; t-butyldimethylsilyl; t-butyldiphenylsilyl; tribenzylsilyl; triphenylsilyl; and diphenylmethylsilyl);Carbonate protecting groups (e.g., methyl, methoxymethyl, 9-fluorenylmethyl; ethyl; 2,2,2-trichloroethyl; 2-(trimethylsilyl)ethyl; vinyl, allyl, nitrophenyl; benzyl; methoxybenzyl; 3,4-dimethoxybenzyl; and nitrobenzyl); carbonyl protecting groups (e.g., acetal and ketal groups, such as dimethyl acetal, 1,3-dioxolane, and the like; acylal groups; and dithiane groups, such as 1,3-dithiane, 1,3-dithiolane, and the like); carboxylic acid protecting groups (e.g., ester groups, such as methyl ester, benzyl ester, t-butyl ester, orthoester, and the like; and oxazoline groups);

[0245] The term "oxo" refers to =O.

[0246] The term "perfluoroalkyl" refers to an alkyl group, as defined herein, in which each hydrogen radical bonded to the alkyl group is replaced by a fluoride radical. Perfluoroalkyl groups are exemplified by trifluoromethyl, pentafluoroethyl, and the like.

[0247] The term "perfluoroalkoxy" refers to an alkoxy group, as defined herein, where each hydrogen radical bonded to the alkoxy group is replaced by a fluoride radical. Perfluoroalkoxy groups are exemplified by trifluoromethoxy, pentafluoroethoxy, and the like.

[0248] The term "spirocyclyl" refers to the C 2-7 Alkyl diradicals, both ends of which are attached to the same carbon atom of the parent group to form a spirocyclyl group, and similarly C 1-6"(C)" refers to a heteroalkyl diradical, both ends of which are bonded to the same atom. The heteroalkyl radical forming the spirocyclyl group can include 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. In some embodiments, the spirocyclyl group contains 1 to 7 carbon atoms, in addition to the carbon atom to which the diradical is attached. The spirocyclyl groups of the present invention can be optionally substituted with 1, 2, 3, or 4 substituents provided herein as optional substituents for the cycloalkyl and / or heterocyclyl groups.

[0249] The term "stereoisomer" refers to all possible different isomeric and conformational forms that a compound (e.g., a compound of any formula described herein) may possess, in particular, all possible stereochemical and conformational isomers, all diastereomers, enantiomers and / or conformational isomers of the basic molecular structure. Some compounds of the present invention can exist in various tautomeric forms, and all isomers are included within the scope of the present invention.

[0250] The term "sulfoalkyl" refers to an alkyl group, as defined herein, substituted with a sulfo group of -SO3H. In some embodiments, the alkyl group can be further substituted with 1, 2, 3, or 4 substituents as described herein, and the sulfo group can be further substituted with one or more O-protecting groups (e.g., as described herein).

[0251] The term "sulfonyl" refers to the group -S(O)2-.

[0252] The term "thioalkylaryl" refers to a chemical substituent of formula -SR, where R is an alkylaryl group. In some embodiments, the alkylaryl group may be further substituted with 1, 2, 3, or 4 substituents as described herein.

[0253] The term "thioalkylheterocyclyl" refers to a chemical substituent of formula -SR, where R is an alkylheterocyclyl group. In some embodiments, the alkylheterocyclyl group can be further substituted with 1, 2, 3, or 4 substituents as described herein.

[0254] The term "thioalkoxy" refers to a chemical substituent of formula -SR, where R is an alkyl group as defined herein. In some embodiments, the alkyl group may be further substituted with 1, 2, 3, or 4 substituents as described herein.

[0255] "Compound": As defined herein, the term "compound" is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted.

[0256] The compounds described herein can be asymmetric (e.g., having one or more stereocenters). Unless otherwise specified, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds of the present disclosure containing asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically active starting materials are well known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described, and the isomers can be isolated as a mixture of isomers or as separated isomers.

[0257] The compounds of the present disclosure also include tautomers. Tautomers result from the interchange of a single bond with an adjacent double bond, accompanied by the migration of a proton. Tautomers include prototropic tautomers, which are protonated isomeric states with the same empirical formula and total charge. Exemplary prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than one site on a heterocyclic ring structure, such as 1H- and 3H-imidazole, 1H-, 2H-, and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomers may exist in equilibrium or may be sterically locked into one form by appropriate substitution.

[0258] The compounds of the present disclosure also include all isotopes of atoms occurring in the intermediates or final compounds. "Isotopes" refer to atoms having the same atomic number but different mass numbers due to the different number of neutrons in their nuclei. For example, isotopes of hydrogen include tritium and deuterium.

[0259] The compounds and salts of the present disclosure can be prepared by routine methods in combination with solvent or water molecules to form solvates and hydrates.

[0260] The term "cyclic" refers to the presence of a continuous ring-like structure. Cyclic molecules need not be circular, but may be linked to form an unbroken chain of subunits. Cyclic molecules, such as the mRNAs of the present invention, may be single units or multimers, and may comprise one or more components of a complex structure or higher order structure.

[0261] "Delivery" refers to the act or method of delivering a compound, substance, entity, component, cargo, or payload.

[0262] "Delivery agent" refers to any substance that at least partially facilitates the delivery of a polynucleotide to a target cell in vivo.

[0263] A "detectable label" refers to one or more markers, signals, or moieties attached to, incorporated into, or associated with another entity that are readily detected by methods known in the art, including x-ray examination, fluorescence, chemiluminescence, enzymatic activity, absorbance, and the like. Detectable labels include radioisotopes, fluorophores, chromophores, enzymes, dyes, metal ions, ligands such as biotin, avidin, streptavidin, and haptens, quantum dots, and the like. The detectable label can be located at any position in the peptides or proteins disclosed herein. The label can be internal to an amino acid, peptide, or protein, or at the N- or C-terminus.

[0264] The term "engineered" refers to a molecule that is designed to have characteristics or properties that differ, whether structurally or chemically, from the starting wild-type or naturally occurring molecule.

[0265] "Expression" of a nucleic acid sequence refers to one or more of the following events: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of the RNA transcript (e.g., by splicing, editing, 5' capping, and / or 3' end processing); (3) translation of the RNA into a polypeptide or protein; and (4) post-translational modification of the polypeptide or protein.

[0266] The term "characteristic" refers to a quality, characteristic, or distinctive element.

[0267] "Fragment," as defined herein, refers to a portion. For example, a fragment of a protein can comprise a polypeptide obtained by digestion of a full-length protein isolated from a cultured cell.

[0268] "Functional": As defined herein, a "functional" biomolecule is a biomolecule in a form in which the molecule exhibits a property and / or activity that characterizes the molecule.

[0269] The phrase "inhibiting gene expression" refers to causing a reduction in the amount of the gene's expression product. The expression product may be RNA (e.g., mRNA) transcribed from the gene, or a polypeptide translated from the mRNA transcribed from the gene. Typically, a reduction in the level of mRNA results in a reduction in the level of the polypeptide translated from the mRNA. The level of expression can be determined using standard techniques for measuring mRNA or protein.

[0270] The term "in vitro" refers to events that take place not in an organism (e.g., an animal, plant, or microorganism) but in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, in a Petri dish, etc.

[0271] The term "in vivo" refers to events that take place within an organism (e.g., an animal, plant, or microorganism, or cells or tissues thereof).

[0272] The term "isolated" refers to a substance or entity that has been separated from at least some of the components with which it was associated (whether in nature or in an experimental setting). Isolated substances can have various levels of purity with respect to the materials with which it was associated. Isolated substances and / or entities can be separated from at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or more of the other components with which they were originally associated. In some embodiments, the isolated agent is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or greater than about 99% pure. As defined herein, a substance is "pure" when it is substantially free of other components. "Substantially isolated": "Substantially isolated" refers to a compound that is substantially (or substantially) separated from the environment in which it was formed or detected. Partial separation includes, for example, compositions enriched for a compound of the present disclosure. Substantial separation includes compositions that contain at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of a compound of the present disclosure or a salt thereof. Methods for isolating compounds and salts thereof are routine in the art.

[0273] The term "linker" refers to a group of atoms (e.g., 10 to 1,000 atoms) and may be composed of atoms or groups such as, but not limited to, carbon, amino, alkylamino, oxygen, sulfur, sulfoxide, sulfonyl, carbonyl, and imine. The linker can be attached at one end to a modified nucleoside or nucleotide, such as a nucleobase or sugar moiety, and at the second end to a payload (e.g., a detectable or therapeutic agent). The linker should be of sufficient length so as not to interfere with incorporation into a nucleic acid sequence. Linkers can be used for any useful purpose, such as to form multimers (e.g., by linking two or more polynucleotides) or complexes and to administer payloads as described herein. Examples of chemical groups that can be incorporated into a linker include, but are not limited to, alkyl, alkenyl, alkynyl, amido, amino, ether, thioether, ester, alkyl, heteroalkyl, aryl, or heterocyclyl, each of which can be optionally substituted as described herein. Examples of linkers include, but are not limited to, unsaturated alkanes, polyethylene glycols (e.g., ethylene glycol or propylene glycol monomer units, e.g., diethylene glycol, dipropylene glycol, triethylene glycol, tripropylene glycol, tetraethylene glycol, or tetraethylene glycol), and dextran polymers. Other examples include, but are not limited to, cleavage moieties within the linker that can be cleaved using reducing agents or photolysis, such as disulfide bonds (-SS-) or azo bonds (-N=N-). Non-limiting examples of selectively cleavable bonds include, for example, amide bonds cleavable by the use of tris(2-carboxyethyl)phosphine (TCEP) or other reducing agents and / or photolysis, and ester bonds cleavable by, for example, acidic or basic hydrolysis.

[0274] "Modified" refers to an altered state or structure of a molecule of the invention. Molecules can be modified in numerous ways, including chemically, structurally, and functionally. In one embodiment, an mRNA molecule of the invention is modified by introducing non-natural nucleosides and / or nucleotides, e.g., as with the natural ribonucleotides A, U, G, and C. Unorthodox nucleotides, such as cap structures, are not considered "modified" even though they differ in chemical structure from the A, C, G, and U ribonucleotides.

[0275] "Naturally occurring" means existing in nature without artificial aid.

[0276] The term "operably linked" refers to a functional connection between two or more molecules, constructs, transcripts, entities, moieties, and the like.

[0277] The term "patient" refers to a subject who may be seeking or in need of treatment, who is in need of treatment, who is receiving treatment, who is going to receive treatment, or who is receiving care from a skilled professional for a particular disease or condition.

[0278] The phrase "optionally substituted X" (e.g., optionally substituted alkyl) is intended to be equivalent to "X, where X is optionally substituted" (e.g., "alkyl, where the alkyl is optionally substituted"). It is not intended to indicate that the subject "X" (e.g., alkyl) itself is optional. Whenever a group is described as "optionally substituted," the group may be unsubstituted or substituted with one or more of the indicated substituents. Similarly, when a group is described as "unsubstituted or substituted," and if substituted, the substituents may be selected from one or more of the indicated substituents. If no substituents are indicated, it means that the indicated "optionally substituted" or "substituted" group can be substituted with one or more group(s) individually and independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, aryl(alkyl), heteroaryl(alkyl), heterocyclyl(alkyl), hydroxy, alkoxy, acyl, cyano, halogen, thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanate, isothiocyanate, azido, nitro, silyl, sulfenyl, sulfinyl, sulfonyl, haloalkyl, haloalkoxy, trihalomethanesulfonyl, trihalomethanesulfonamido, amino, monosubstituted amino, and disubstituted amino.

[0279] A "peptide" is 50 amino acids or less in length, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids in length.

[0280] The phrase "pharmaceutically acceptable" is used herein to refer to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0281] The phrase "pharmaceutically acceptable excipient," as defined herein, refers to any ingredient other than the compounds described herein (e.g., a vehicle capable of suspending or dissolving an active compound) that has the properties of being substantially non-toxic and non-inflammatory to a patient. Excipients include, for example, anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colorants), emollients, emulsifiers, fillers (diluents), film formers or coating agents, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and water for hydration. Exemplary excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0282] The present disclosure also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds, in which the parent compound is modified by converting an existing acid or base moiety into its salt form (for example, by reacting a free base group with a suitable organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues, such as amines; alkali or organic salts of acidic residues, such as carboxylic acids; etc. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, bromate, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lanthanide ... Examples of salts that can be used include butobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and valerate.Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations, such as, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, salts with organic bases (e.g., organic amines), such as benzathine, dicyclohexylamine, hydrabamine (formed with N,N-bis(dehydroabietyl)ethylenediamine), N-methyl-D-glucamine, N-methyl-D-glucamide, t-butylamine, and salts with amino acids such as arginine or lysine. Basic nitrogen-containing groups may be quaternized with materials such as lower alkyl halides (e.g., methyl, ethyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, and dibutyl sulfate), long chain halides (e.g., decyl chloride, bromide, and iodide, lauryl, and stearyl), aryl alkyl halides (e.g., benzyl and phenethyl bromides), and the like.

[0283] The pharmaceutically acceptable salts of the present disclosure include the conventional non-toxic salts of parent compounds, for example, formed from non-toxic inorganic or organic acids.The pharmaceutically acceptable salts of the present disclosure can be synthesized from parent compounds that contain a basic or acidic moiety by conventional chemical methods.Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of suitable base or acid in water or organic solvent, or in the mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418; Pharmaceutical Salts: Properties, Selection, and Use, P.H. Stahl and C.G. Wermuth (eds.), Wiley-VCH, 2008; and Berge et al., Journal of Pharmaceutical Science, 66, 1-19 (1977); P. Gould, International J. Pharmaceutics (1986) 33, 201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and The Orange Book (Food & Drug Administration, Washington, DC, on their website), the disclosures of which are each incorporated herein by reference in their entirety.

[0284] All such acid and base salts are pharmaceutically acceptable salts within the scope of this invention, and all acid and base salts shall be considered equivalent to the free form of the corresponding compound of Formula I for purposes of this disclosure.

[0285] "Pharmacokinetics" refers to any one or more properties of a molecule or compound that are relevant to determining the fate of an administered substance in an organism. Pharmacokinetics is divided into several areas, including the extent and rate of absorption, distribution, metabolism, and excretion. This is commonly referred to as ADME: (A) Absorption is the process by which a substance enters the blood circulation; (D) Distribution is the dispersion or dispersal of a substance throughout the body's fluids and tissues; (M) Metabolism (or biotransformation) is the irreversible conversion of a parent compound to daughter metabolites; and (E) Excretion (or elimination) refers to the removal of a substance from the body. In rare cases, some pharmaceuticals irreversibly accumulate in body tissues.

[0286] The term "pharmaceutically acceptable solvate" refers to a compound of the present invention in which molecules of a suitable solvent are incorporated into the crystal lattice. A suitable solvent is physiologically tolerable at the administered dose. For example, solvates can be prepared by crystallization, recrystallization, or precipitation from a solution containing an organic solvent, water, or a mixture thereof. Examples of suitable solvents include ethanol, water (e.g., monohydrate, dihydrate, and trihydrate), N-methylpyrrolidinone (NMP), dimethyl sulfoxide (DMSO), N,N'-dimethylformamide (DMF), N,N'-dimethylacetamide (DMAC), 1,3-dimethyl-2-imidazolidinone (DMEU), 1,3-dimethyl-3,4,5,6-tetrahydro-2-(1H)-pyrimidinone (DMPU), acetonitrile (ACN), propylene glycol, ethyl acetate, benzyl alcohol, 2-pyrrolidone, benzyl benzoate, and the like. When water is the solvent, the solvate is referred to as a "hydrate."

[0287] "Physicochemical" refers to physical and / or chemical properties.

[0288] The term "preventing" refers to at least one of: partially or completely delaying the onset of an infection, disease, disorder, and / or condition; partially or completely delaying the onset of one or more symptoms, characteristics, or clinical signs of a particular infection, disease, disorder, and / or condition; partially or completely delaying the onset of one or more symptoms, characteristics, or signs of a particular infection, disease, disorder, and / or condition; partially or completely delaying the progression of an infection, a particular disease, disorder, and / or condition; and reducing the risk of developing pathology associated with an infection, disease, disorder, and / or condition.

[0289] The term "protein of interest" or "desired protein" includes those provided herein as well as fragments, mutants, variants and modifications thereof.

[0290] "Purify," "purified," and "purification" refer to making substantially pure or clear from unwanted components, contaminating materials, impurities, or imperfections.

[0291] The term "sample" or "biological sample" refers to a subset of tissues, cells, or component parts thereof (e.g., bodily fluids, including but not limited to, blood, mucus, lymph, synovial fluid, cerebrospinal fluid, saliva, amniotic fluid, amniotic cord blood, urine, vaginal fluid, and semen). Samples can also include homogenates, lysates, or extracts prepared from whole organisms or a subset of their tissues, cells, or component parts, or fractions or portions thereof, including but not limited to, plasma, serum, cerebrospinal fluid, lymph, external portions of the skin, respiratory, intestinal, and genitourinary tracts, tears, saliva, milk, blood cells, tumors, and organs. Samples can also refer to media, e.g., nutrient broths or gels, which may contain cellular components such as proteins or nucleic acid molecules.

[0292] The terms "prominent" or "prominently" are used synonymously with the term "almost, almost."

[0293] A "single unit dose" is a dose of any therapeutic agent administered in one dose / at one time / by a single route / at a single point of contact (i.e., a single administration event).

[0294] A "split dose" is the division of a single unit dose or total daily dose into two or more doses.

[0295] "Stable" refers to a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and preferably amenable to formulation into an efficacious therapeutic agent.

[0296] The term "subject" or "patient" refers to any organism to which a composition according to the invention may be administered, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Exemplary subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans) and / or plants.

[0297] The term "approximately" refers to a qualitative state indicating the total or nearly total extent or degree of a quality or characteristic of interest. As will be understood by those skilled in the art of biology, biological and chemical phenomena often do not, if ever, proceed to completion and / or perfection or achieve or escape an absolute result. Thus, the term "approximately" is used herein to capture the potential lack of inherent perfection in many biological and chemical phenomena.

[0298] The term "approximately equal" refers to the time difference between doses, and the term refers to plus / minus 2%.

[0299] The term "approximately simultaneously" refers to multiple administrations, and the term refers to within 2 seconds.

[0300] An individual "suffering from" a disease, disorder, and / or condition has been diagnosed with or exhibits one or more symptoms of the disease, disorder, and / or condition.

[0301] An individual who is "susceptible" to a disease, disorder, and / or condition may not have been diagnosed with and / or may not exhibit symptoms of the disease, disorder, and / or condition, but has a tendency to develop the disease or its symptoms. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., cancer) may be characterized by one or more of the following: (1) a genetic mutation associated with the development of the disease, disorder, and / or condition; (2) a genetic polymorphism associated with the development of the disease, disorder, and / or condition; (3) an increase and / or decrease in the expression and / or activity of a protein and / or nucleic acid associated with the disease, disorder, and / or condition; (4) habits and / or lifestyle associated with the development of the disease, disorder, and / or condition; (5) a family history of the disease, disorder, and / or condition; and (6) exposure to and / or infection by a microorganism associated with the development of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will likely develop the disease, disorder, and / or condition, hi some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will not develop the disease, disorder, and / or condition.

[0302] The term "synthetic" refers to produced, prepared, and / or manufactured by the hand of man. Synthesis of polynucleotides or polypeptides or other molecules of the invention can be chemical or enzymatic.

[0303] The term "target cell" refers to any one or more cells of interest. Cells may be found in vitro, in vivo, in situ, or within a tissue or organ of an organism. The organism may be an animal, preferably a mammal, more preferably a human, and most preferably a patient.

[0304] The term "therapeutic agent" refers to any agent that, when administered to a subject, has a therapeutic, diagnostic, and / or prophylactic effect and / or induces a desired biological and / or pharmacological effect.

[0305] The term "therapeutically effective amount" refers to the amount of an agent (e.g., nucleic acid, drug, therapeutic agent, diagnostic agent, prophylactic agent, etc.) to be delivered that is sufficient to treat, ameliorate symptoms of, diagnose, prevent, and / or delay the onset of the infection, disease, disorder, and / or condition when administered to a subject suffering from or susceptible to the infection, disease, disorder, and / or condition.

[0306] The term "therapeutically effective outcome" refers to an outcome sufficient to treat, ameliorate, diagnose, prevent, and / or delay the onset of an infection, disease, disorder, and / or condition in a subject suffering from or susceptible to the infection, disease, disorder, and / or condition.

[0307] The phrase "total daily dose" refers to the amount given or prescribed during a 24-hour period. It may be administered as a single unit dose.

[0308] The term "treating" refers to the partial or complete alleviation, amelioration, improvement, relief, delay in onset, inhibition of progression, reduction in severity, and / or reduction in incidence of one or more symptoms or characteristics of a particular infection, disease, disorder, and / or condition. For example, "treating" cancer can refer to inhibiting tumor survival, growth, and / or metastasis. Treatment may be administered to subjects who do not exhibit signs of the disease, disorder, and / or condition, as well as to subjects who exhibit only early signs of the disease, disorder, and / or condition, with the intent of reducing the risk of developing pathology associated with the disease, disorder, and / or condition.

[0309] "Unmodified" refers to any substance, compound, or molecule before it has been altered in any way. Unmodified can, but does not always, refer to the wild-type or native form of a biomolecule. A molecule can undergo a series of modifications, whereby each modified molecule can serve as the "unmodified" starting molecule for subsequent modifications.

[0310] The term "antisense nucleic acid" refers to a non-enzymatic nucleic acid molecule that binds to a target RNA through RNA-RNA, RNA-DNA, or RNA-PNA (protein nucleic acid; Egholm et al., 1993 Nature 365, 566) interactions and alters the activity of the target RNA (for reviews, see Stein and Cheng, 1993 Science 261, 1004, and Woolf et al., U.S. Pat. No. 5,849,902). Typically, an antisense molecule is complementary to a target sequence along a single, contiguous sequence of the antisense molecule. However, in certain embodiments, an antisense molecule can bind to a substrate such that the substrate molecule forms a loop, and / or an antisense molecule can bind to a substrate such that the antisense molecule forms a loop. Thus, an antisense molecule may be complementary to two (or more) non-contiguous substrate sequences, or two (or more) non-contiguous sequence portions of the antisense molecule may be complementary to a target sequence, or both. Furthermore, antisense DNA can be used to target RNA through DNA-RNA interactions, thereby activating RNase H, which digests the target RNA in the duplex. Antisense oligonucleotides can contain one or more RNAse H-activating regions that can activate RNAse H cleavage of the target RNA. Antisense DNA can be chemically synthesized or expressed using a single-stranded DNA expression vector or its equivalent. "Antisense RNA" is an RNA strand with a sequence complementary to a target gene mRNA and can induce RNAi by binding to the target gene mRNA. "Antisense RNA" is an RNA strand with a sequence complementary to a target gene mRNA and is thought to induce RNAi by binding to the target gene mRNA. "Sense RNA" has a sequence complementary to the antisense RNA and anneals to its complementary antisense RNA to form an iNA. These antisense and sense RNAs have traditionally been synthesized using an RNA synthesizer.

[0311] The term "nucleic acid" refers to deoxyribonucleotides or ribonucleotides and polymers thereof in single- or double-stranded form. The term encompasses synthetic, natural, and non-natural nucleic acids containing known nucleotide analogs or modified backbone residues or linkages that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to the reference nucleotide. Examples of such analogs include, but are not limited to, phosphorothioates, phosphoramidates, methyl phosphonates, chiral methyl phosphonates, 2'-O-methyl ribonucleotides, and peptide nucleic acids (PNAs).

[0312] The term "RNA" refers to a molecule containing at least one ribonucleotide residue. "Ribonucleotide" refers to a nucleotide having a hydroxyl group at the 2' position of a β-D-ribofuranose moiety. This term includes double-stranded RNA, single-stranded RNA, isolated RNA, e.g., partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, and modified RNA that differs from naturally occurring RNA by the addition, deletion, substitution, and / or modification of one or more nucleotides. Such modifications can include the addition of non-nucleotide material, for example, to the end(s) of an interfering RNA or internally, e.g., at one or more nucleotides of the RNA. Nucleotides in the RNA molecules of the present disclosure can also include non-standard nucleotides, e.g., non-naturally occurring nucleotides, or chemically synthesized nucleotides, or deoxynucleotides. These modified RNAs can be referred to as analogs or analogs of naturally occurring RNA. As used herein, the terms "ribonucleic acid" and "RNA" refer to molecules containing at least one ribonucleotide residue, including siRNA, antisense RNA, single-stranded RNA, microRNA, mRNA, non-coding RNA, and polyvalent RNA. A ribonucleotide is a nucleotide with a hydroxyl group at the 2' position of a β-D-ribofuranose moiety. These terms include double-stranded RNA, single-stranded RNA, isolated RNA, e.g., partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, and modified and altered RNA that differs from natural RNA by the addition, deletion, substitution, modification, and / or alteration of one or more nucleotides. RNA modifications can include the addition of non-nucleotide material, for example, at one or more nucleotides of the RNA (nucleotides in RNA molecules include non-standard nucleotides, e.g., non-natural nucleotides, or chemically synthesized nucleotides, or deoxynucleotides), for example, to the end of an interfering RNA or within the RNA. These modified RNAs can be referred to as analogs.

[0313] The term "nucleotide" refers to natural (standard) bases and modified bases known in the art. Such bases are generally located at the 1' position of the sugar moiety of the nucleotide. A nucleotide generally comprises a base, a sugar, and a phosphate group. A nucleotide can be unmodified or modified at the sugar, phosphate, and / or base moieties (which may also be interchangeably referred to as a nucleotide analog, modified nucleotide, non-natural nucleotide, non-standard nucleotide, etc.; see, e.g., Usman and McSwiggen, supra; Eckstein, et al., International PCT Publication No. WO 92 / 07065; Usman, et al., International PCT Publication No. WO 93 / 15187; Uhlman & Peyman, supra, all of which are incorporated herein by reference). Several examples of modified nucleobases are known in the art, as summarized in Limbach, et al., Nucleic Acids Res. 22:2183, 1994. Non-limiting examples of base modifications that can be introduced into nucleic acid molecules include inosine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxybenzene, 3-methyluracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidine (e.g., 5-methylcytidine), 5-alkyluridine (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine), or 6-azapyrimidine or 6-alkylpyrimidine (e.g., 6-methyluridine), propyne, etc. (Burgin, et al., Biochemistry 35:14090, 1996; Uhlman & Peyman, supra). In this embodiment, a "modified base" refers to a nucleotide base other than adenine, guanine, cytosine, and uracil at the 1' position, or an equivalent thereof.

[0314] The phrase "complementary nucleotide bases" refers to a pair of nucleotide bases that form hydrogen bonds with each other. In RNA, adenine (A) pairs with thymine (T) or uracil (U), and guanine (G) pairs with cytosine (C). Complementary segments or strands of nucleic acid hybridize (join by hydrogen bonds) with each other. "Complementarity" refers to the ability of a nucleic acid to form hydrogen bonds with another nucleic acid sequence via traditional Watson-Crick or other non-traditional bonding modes.

[0315] The term "microRNA (miRNA)" refers to single-stranded RNA molecules approximately 21–23 nucleotides in length that regulate gene expression. miRNAs are encoded by genes transcribed from DNA but are not translated into proteins (non-coding RNAs); instead, they are processed from primary transcripts known as pri-miRNAs into short stem-loop structures called pre-miRNAs and ultimately into functional miRNAs. Mature miRNA molecules are partially complementary to one or more messenger RNA (mRNA) molecules, and their primary function is to downregulate gene expression.

[0316] The terms "small interfering RNA (siRNA)," "short interfering RNA," and "silencing RNA" refer to a group of double-stranded RNA molecules, 16-40 nucleotides in length, that serve a variety of roles in biology. Most notably, siRNAs participate in the RNA interference (RNAi) pathway, where they interfere with the expression of specific genes. In addition to their role in the RNAi pathway, siRNAs also function in RNAi-related pathways, for example, as antiviral mechanisms or in shaping the chromatin structure of the genome. The complexity of these pathways is only just beginning to be elucidated.

[0317] The term "RNAi" refers to the RNA-dependent gene silencing process, controlled by the RNA-induced silencing complex (RISC), initiated by short double-stranded RNA molecules within the cell that interact with the catalytic RISC component Argonaute. When double-stranded RNA or RNA-like iNAs or siRNAs are exogenous (arising from infection with a virus carrying an RNA genome or transfected iNAs or siRNAs), the RNA or iNAs are directly taken up into the cytoplasm and cleaved into short fragments by the enzyme Dicer. Alternatively, the initiating dsRNA may be endogenous (derived from the cell), such as pre-microRNAs expressed from RNA-coding genes within the genome. The primary transcript from such genes is first processed to form the characteristic stem-loop structure of pre-miRNAs in the nucleus, then transported to the cytoplasm where it is cleaved by Dicer. Thus, the two dsRNA pathways, exogenous and endogenous, converge within the RISC complex. The active components of the RNA-induced silencing complex (RISC) are endonucleases called Argonaute proteins, which cleave the target mRNA strand complementary to the bound siRNA or iNA. Because the fragments generated by Dicer are double-stranded, they can theoretically generate functional siRNA or iNA, respectively. However, only one of the two strands, known as the guide strand, binds to the Argonaute protein and directs gene silencing. The other, non-guide strand, or passenger strand, is degraded upon RISC activation.

[0318] The term "miRNA mimic" refers to chemically modified double-stranded RNAs that mimic endogenous miRNAs and allow for analysis of miRNA function by upregulating miRNA activity.

[0319] The phrase "commercially available compound" and the compounds used in the examples herein may be obtained from conventional commercial sources, such as Acros Organics (Pittsburgh, Pa.), Sigma-Adrich Chemical (Milwaukee, Wis.), Avocado Research (Lancashire, UK), Bionet (Cornwall, UK), Boron Molecular (Research Triangle Park, NC), Combi-Blocks (San Diego, Calif.), Eastman Organic Chemicals, Eastman Kodak Company (Rochester, NY), Fisher Scientific Co. (Pittsburgh, Pa.), Frontier Scientific (Logan, Utah), ICN Biomedicals, Inc. (Costa Mesa, Calif.), Lancaster Synthesis (Windham, NH), Maybridge Chemical Co. (Cornwall, UK), Pierce Chemical Co. (Rockford, Ill.), and Riedel de Haen (Hannover, Germany), Spectrum Quality Products, Inc. (New Brunswick, NJ), TCI America (Portland, Oreg.), and Wako Chemicals USA, Inc. (Richmond, Va.).

[0320] The phrase "a compound described in the chemical literature" refers to a compound that may be identified through reference books and databases covering chemical compounds and chemical reactions known to those skilled in the art. Suitable reference books and treatises that detail the synthesis of reactants useful for preparing the compounds disclosed herein or that refer to articles that describe the preparation of the compounds disclosed herein include, for example, "Synthetic Organic Chemistry," John Wiley and Sons, Inc. New York; S.R. Sandler et al., "Organic Functional Group Preparations," 2nd Ed., Academic Press, New York, 1983; H.O. House, "Modern Synthetic Reactions," 2nd Ed., W.A. Benjamin, Inc. Menlo Park, Calif., 1972; T.L. Glichrist, "Heterocyclic Chemistry," 2nd Ed., John Wiley and Sons, New York, 1992; J. March, "Advanced Organic Chemistry: reactions, Mechanisms and Structure," 5th Ed., Wiley Interscience, New York, 2001. Specific reactants and similar reactants may be identified through indexes of known compounds prepared by the American Chemical Society's Chemical Abstract Service, available in most public and university libraries, or through online databases (for more information, contact the American Chemical Society, Washington, DC). Compounds that are known but cannot be purchased from a catalog may be prepared by custom chemical synthesis houses, many of which standard chemical supply houses (e.g., the sources listed above) offer custom synthesis services.

[0321] The term "lipid" refers to organic compounds that contain esters of fatty acids and are characterized by being insoluble in water but soluble in many organic solvents. Lipids are generally divided into at least three classes: (1) "simple lipids," which include fats and oils, as well as waxes; (2) "complex lipids," which include phospholipids and glycolipids; and (3) "derived lipids," such as steroids.

[0322] The term "lipid particle" refers to a lipid formulation that can be used to deliver a therapeutic nucleic acid (e.g., mRNA) to a target site of interest (e.g., a cell, tissue, organ, etc.). In a preferred embodiment, the lipid particle is a nucleic acid-lipid particle typically formed from a cationic lipid, a non-cationic lipid (e.g., a phospholipid), a conjugated lipid that prevents particle aggregation (e.g., a PEG-lipid), and optionally cholesterol. Typically, the therapeutic nucleic acid (e.g., mRNA) is encapsulated in the lipid portion of the particle, thereby protecting it from enzymatic degradation.

[0323] The lipid particles typically have an average diameter of 30 nm to 150 nm, 40 nm to 150 nm, 50 nm to 150 nm, 60 nm to 130 nm, 70 nm to 110 nm, 70 nm to 100 nm, 80 nm to 100 nm, 90 nm to 100 nm, 70 to 90 nm, 80 nm to 90 nm, 70 nm to 80 nm, or 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm, and are substantially non-toxic. In addition, nucleic acids, when present in lipid particles of the present disclosure, are resistant in aqueous solution to degradation by nucleases.

[0324] "Solvate" refers to a physical association of a compound of the present disclosure with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate can be isolated. "Solvate" encompasses both solution-phase and isolable solvates. Non-limiting examples of suitable solvates include ethanolates, methanolates, and the like.

[0325] The term "lipid-encapsulated" refers to lipid particles that provide a therapeutic nucleic acid, such as mRNA, fully encapsulated, partially encapsulated, or both. In preferred embodiments, the nucleic acid (e.g., mRNA) is fully encapsulated in the lipid particle.

[0326] "Lipid complex" refers to a complex lipid that inhibits lipid particle aggregation. Such lipid complexes include, but are not limited to, PEG-lipid complexes, such as PEG conjugated to dialkyloxypropyl (e.g., PEG-DAA complexes), PEG conjugated to diacylglycerol (e.g., PEG-DAG complexes), PEG conjugated to cholesterol, PEG conjugated to phosphatidylethanolamine, and PEG conjugated to ceramide, cationic PEG-lipid, polyoxazoline (POZ)-lipid complexes, polyamide oligomers, and mixtures thereof. PEG or POZ can be directly conjugated to lipids or linked to lipids via a linker moiety. For example, any linker moiety suitable for linking PEG or POZ to lipids can be used, including non-ester-containing linker moieties and ester-containing linker moieties. In certain preferred embodiments, non-ester-containing linker moieties such as amides or carbamates are used.

[0327] The term "amphipathic lipid" refers to a lipid material in which the hydrophobic portion of the lipid material orients within the hydrophobic phase, while the hydrophilic portion orients toward the aqueous phase. The hydrophilic character comes from the presence of polar or charged groups, such as carbohydrates, phosphates, carboxylic acids, sulfato, amino, sulfhydryl, nitro, hydroxyl, and other similar groups. Hydrophobicity can be imparted by the inclusion of nonpolar groups, including, but not limited to, long-chain saturated and unsaturated aliphatic hydrocarbon groups, and such groups substituted with one or more aromatic, alicyclic, or heterocyclic group(s). Examples of amphipathic compounds include, but are not limited to, phospholipids, aminolipids, and sphingolipids.

[0328] Representative examples of phospholipids include, but are not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine, dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, distearoylphosphatidylcholine, and dilinoleoylphosphatidylcholine.Other compounds that do not contain phosphorus, such as sphingolipids, glycosphingolipid family, diacylglycerol, and β-acyloxyacid, are also included in the group designated as amphipathic lipid.In addition, the above amphipathic lipids can be mixed with other lipids, including triglycerides and sterols.

[0329] The term "neutral lipid" refers to any of a number of lipid species that exist in uncharged or zwitterionic form at selected pHs. At physiological pH, such lipids include, for example, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides, and diacylglycerol.

[0330] The term "non-cationic lipid" refers to an amphipathic lipid or a neutral lipid or an anionic lipid, and is described in more detail below.

[0331] The term "anionic lipid" refers to any lipid that has a negative charge at physiological pH, including, but not limited to, phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoylphosphatidylethanolamine, N-succinylphosphatidylethanolamine, N-glutarylphosphatidylethanolamine, lysylphosphatidylglycerol, palmitoyloleoylphosphatidylglycerol (POPG), and other anionic modifying groups attached to neutral lipids.

[0332] The term "hydrophobic lipid" refers to a compound having a non-polar group, including but not limited to, long-chain saturated and unsaturated aliphatic hydrocarbon groups, and those groups optionally substituted with one or more aromatic, alicyclic or heterocyclic groups. Suitable examples include, but are not limited to, diacylglycerol, dialkylglycerol, N-N-dialkylamino, 1,2-diacyloxy-3-aminopropane and 1,2-dialkyl-3-aminopropane.

[0333] The terms "cationic lipid" and "amino lipid" are used interchangeably and refer to those lipids and their salts having one, two, three, or more fatty acid or fatty alkyl chains and a pH-titratable amino head group (e.g., alkylamino or dialkylamino head group). Cationic lipids are typically protonated (i.e., positively charged) at a pH below the pKa of the cationic lipid and substantially neutral at a pH above the pKa. The cationic lipids of the present disclosure may also be referred to as titratable cationic lipids. In some embodiments, cationic lipids are C 2 -C 3 -C 4 -C 5 -C 6 -C 7 -C 8 -C 9 -C 10 -C 11 -C 12 -C 13 -C 14 -C 15 -C 16 -C 17 -C 18 -C 19 -C 20 -C 21 -C 22 -C 23 -C 24 -C 25 -C 26 -C 27 -C 28 -C 29 -C 30 -C 31 -C 32 -C 33 -C 34 -C 35 -C 40 -C 16 -C 29 -C 35 -C 17 -C 28 -C 18 -C 29 -C 35 -C 29 -C 36 -C 18 -C 21 -C 22 -C 23 -C 36 -C 19 -C 24 -C 25 -C 26 -C 27 -C 28 -C 29 -C 30 -C 31 -C 29 -C 32 -C 33 -C 34 -C 35 -C 16 -C 29 -C 35 -C 17 -C 29 -C 35 -C 18 -C 29 -C 35 -C 19 -C 21 -C 2 18Cationic lipids include, but are not limited to, DSDMA, DODMA, DLinDMA, DLenDMA, γ-DLenDMA, DLin-K-DMA, DLin-K-C2-DMA (also known as DLin-C2K-DMA, XTC2, and C2K), DLin-K-C3-DMA, DLin-K-C4-DMA, DLen-C2K-DMA, y-DLen-C2K-DMA, DLin-M-C2-DMA (also known as MC2), DLin-M-C3-DMA (also known as MC3), and (DLin-MP-DMA) (also known as 1-Bl 1).

[0334] "Substituted" means substitution with a specified group other than hydrogen, or with one or more groups, moieties, or radicals which may be the same or different, each eg, independently selected.

[0335] Equivalents and Scope Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments in accordance with the present disclosure described herein. The scope of the present disclosure is not intended to be limited to the above description, but is instead set forth in the appended claims.

[0336] It is further noted that the term "comprising" is intended to be open and allows for, but does not require, the inclusion of additional elements or steps. When the term "comprising" is used herein, the term "consisting of" is also included and disclosed.

[0337] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments in accordance with the present disclosure described herein. The scope of the present disclosure is not intended to be limited to the above description, but is instead set forth in the appended claims.

[0338] In addition, it should be understood that any particular embodiment of the present disclosure that falls within the scope of the prior art may be expressly excluded from any one or more claims. Because such embodiments are considered to be well known to those skilled in the art, embodiments may be excluded even if the exclusion is not expressly stated herein. Any particular embodiment of the composition of the present disclosure (e.g., any nucleic acid or protein encoded by the nucleic acid; any production method; any use method, etc.) may be excluded from any one or more claims for any reason, regardless of whether it is related to the existence of prior art.

[0339] All cited sources, e.g., references, publications, databases, database entries, and techniques cited herein, are incorporated herein by reference, even if not expressly stated in the citation. In the event of a conflict between a cited source and this application, the statements in this application shall control.

[0340] Pharmaceutical Composition It should be understood that the compounds disclosed herein optionally include reference to their salts unless otherwise specified. The term "salt(s)," as used herein, refers to salts formed with inorganic and / or organic bases as well as acid salts formed with inorganic and / or organic acids. In addition, when a compound of Formula I contains both a basic moiety, such as, but not limited to, pyridine or imidazole, and an acidic moiety, such as, but not limited to, a carboxylic acid, zwitterions ("internal salts") may be formed and are included in the term "salt(s)" as used herein. While other salts are also useful, pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred. Salts of compounds of Formula I may be formed, for example, by reacting equivalent amounts of a compound of Formula I with a predetermined amount of acid or base in an aqueous medium, from which the salt precipitates or is subsequently lyophilized.

[0341] The compounds disclosed herein can exist in unsolvated and solvated forms, including hydrated forms. In general, the solvated forms, with pharmaceutically acceptable solvents such as water, ethanol, etc., are equivalent to the unsolvated forms for purposes of the present invention.

[0342] Compounds disclosed herein, and salts and solvates thereof, may exist in their tautomeric form (for example, as an amide or imino ether), and all such tautomeric forms are contemplated as being included herein as part of the present invention.

[0343] Polymorphs of the compounds of the present invention are also within the scope of the present invention (ie, polymorphs of the compounds of Formula I are within the scope of this disclosure).

[0344] All stereoisomers (e.g., geometric isomers and optical isomers) of the compounds of the present invention (including salts, solvates, and prodrugs of the compounds, and salts and solvates of the prodrugs) are contemplated within the scope of the present disclosure, including enantiomeric forms (which may exist even when no asymmetric carbon is present), rotamer forms, atropisomers, and diastereomeric forms, such as those that may exist due to asymmetric carbons on various substituents. The chiral centers of the compounds herein may have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of terms such as "salt" and "solvate" is intended to apply equally to enantiomers, stereoisomers, rotamers, tautomers, racemates, or prodrug salts and solvates of the compounds of the present disclosure.

[0345] lipid nanoparticles The compound of Formula I or a pharmaceutically acceptable salt thereof can be contained in a lipid composition comprising nanoparticles or a bilayer of lipid molecules. The lipid bilayer preferably further comprises a neutral lipid or polymer. The lipid composition preferably comprises a liquid medium. The composition preferably further encapsulates a nucleic acid. The nucleic acid is preferably an mRNA encoding a protein or polypeptide of interest and having translation activity to produce a target protein. Alternatively, the nucleic acid is preferably an siRNA that regulates gene expression, preferably inducing gene expression knockdown (i.e., gene silencing). The lipid composition preferably further comprises siRNA and / or mRNA and a neutral lipid or polymer. The lipid composition preferably encapsulates siRNA and / or mRNA.

[0346] In certain embodiments, the lipid nanoparticles described herein comprise four lipid components: a phospholipid; cholesterol; a PEG-lipid; and an ionizable lipid. Preferably, the phospholipid is DSPC, the PEG-lipid is PEG-DMG, and the cationic lipid is a lipid of Formula II, Formula III, or Formula IV. In some embodiments, the molar composition is about 40-70:5:20:20:50:1:10 or 50-60:5:10:30:40:1:5 lipid:DSPC:cholesterol:PEG-DMG. More preferably, the molar composition is about 58:7:33.5:1.5 or 50:7:40:3 lipid:DSPC:cholesterol:PEG-DMG. In certain embodiments, the organic solvent concentration at which the lipid is solubilized is about 45% v / v to about 90% v / v. In certain preferred embodiments, the organic solvent is a lower alkanol. Suitable lower alkanols include, for example, methanol, ethanol, propanol, butanol, pentanol, and isomers and combinations thereof. The solvent is preferably ethanol having a volume of about 50 to 90% v / v. Preferably, the lipid occupies a volume of about 1 mL / g to about 5 mL / g.

[0347] In some embodiments, the siRNA and / or mRNA is fully encapsulated within the lipid portion of the lipid particle such that the siRNA and / or mRNA in the lipid particle is resistant to nuclease degradation in aqueous solution. In other embodiments, the lipid particles described herein are substantially non-toxic to mammals, such as humans. The lipid particles typically have an average diameter of 30 nm to 150 nm, 40 nm to 150 nm, 50 nm to 150 nm, 60 nm to 130 nm, 70 nm to 110 nm, or 70 nm to 90 nm. The lipid particles of the present disclosure also typically have a lipid:RNA ratio (mass / mass) of 1:1 to 100:1, 1:1 to 50:1, 2:1 to 25:1, 3:1 to 20:1, 5:1 to 15:1, or 5:1 to 10:1, or 10:1 to 14:1, or 9:1 to 20:1.

[0348] In a preferred embodiment, the lipid particles comprise siRNA and / or mRNA, an ionizable lipid (e.g., one or more cationic lipids or salts thereof described herein), a phospholipid, and a conjugated lipid (e.g., one or more PEG-lipid conjugates) that inhibit particle aggregation. The lipid particles may also comprise cholesterol. The lipid particles may comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mRNAs that express one or more polypeptides. The lipid particles may comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more siRNAs that induce gene expression knockdown.

[0349] In nucleic acid-lipid particles, siRNA and / or mRNA can be completely encapsulated in the lipid portion of the particle, thereby protecting nucleic acid from nuclease degradation.In a preferred embodiment, the lipid particles containing siRNA and / or mRNA are completely encapsulated in the lipid portion of the particle, thereby protecting nucleic acid from nuclease degradation.In certain cases, the siRNA and / or mRNA in lipid particles are not substantially decomposed after the particles are exposed to nuclease at 37 ℃ for at least 20, 30, 45 or 60 minutes.In certain other cases, the siRNA and / or mRNA in lipid particles are not substantially decomposed after the particles are incubated in serum at 37 ℃ for at least 30, 45 or 60 minutes, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34 or 36 hours. In other embodiments, the siRNA and / or mRNA is complexed with the lipid portion of the particle. One advantage of the formulations of the present disclosure is that the nucleic acid-lipid particle compositions are substantially non-toxic to mammals, such as humans.

[0350] " Fully encapsulated " means that the nucleic acid (for example, siRNA and / or mRNA) in nucleic acid-lipid particles is not significantly degraded after being exposed to serum or nuclease assay, which would significantly degrade free siRNA and / or mRNA.When fully encapsulated, preferably less than 25% of the nucleic acid in the particles is degraded in a treatment that would normally degrade 100% of free nucleic acid, more preferably less than 10%, most preferably less than 5% of the nucleic acid in the particles is degraded." Fully encapsulated " also means that when administered in vivo, nucleic acid-lipid particles do not rapidly decompose into their component parts.

[0351] A "lipid NA nanoparticle" is any lipid composition that can be used to deliver a compound, such as, but not limited to, a liposome comprising a lipid bilayer in either a unilamellar or multilamellar structure, in which the RNA is at least partially encapsulated by ion-pairing with an ionizable lipid.

[0352] "Lamellar morphology" refers to a bilayer structure. The lamellar morphology, bilayer structure of the lipids disclosed herein can be determined using analytical techniques, such as cryo-TEM imaging.

[0353] "Lipid-encapsulated" can refer to a lipid formulation that provides a compound by complete encapsulation, partial encapsulation, or both, where the RNA is inaccessible to RNase-mediated hydrolysis or dye intercalation.

[0354] In the context of nucleic acids, complete encapsulation can be determined by performing a membrane-impermeable fluorescent dye exclusion assay, which uses a dye that exhibits enhanced fluorescence upon association with nucleic acids. Encapsulation is determined by adding the dye to a liposome formulation, measuring the resulting fluorescence, and comparing it to the fluorescence observed upon the addition of a small amount of nonionic detergent. Detergent-mediated disruption of the liposome bilayer releases the encapsulated nucleic acid, allowing it to interact with the membrane-impermeable dye. Nucleic acid encapsulation can be calculated as E = (I0 - I) / I0, where I0 and I0 refer to the fluorescence intensity before and after the addition of detergent.

[0355] In other embodiments, the present disclosure provides siRNA and / or mRNA-lipid particle compositions comprising a plurality of siRNA and / or mRNA-lipid particles.

[0356] The lipid particles contain siRNA and / or mRNA that is fully encapsulated within the lipid portion of the particle, resulting in 30%-100%, 40%-100%, 50%-100%, 60%-100%, 70%-100%, 80%-100%, 90%-100%, 30%-95%, 40%-95%, 50%-95%, 60%-95%, 70%-95%, 80%-95%, 85%-95%, 90%-95%, 30%-90 ... 0%, 50%-90%, 60%-90%, 70%-90%, 80%-90%, or at least 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% (or any percentage or range thereof) of the particles have encapsulated siRNA and / or mRNA therein.

[0357] Depending on the intended use of the compositions disclosed herein, including therapeutic siRNA and / or mRNA molecules and lipid particles, the ratio of components can be varied, and the delivery efficiency of a particular formulation can be measured using assays known in the art.

[0358] cationic lipids The composition may contain cationic lipids suitable for forming cationic liposomes or lipid nanoparticles.Cationic lipids have been widely studied for nucleic acid delivery because they can bind to negatively charged membranes and induce uptake.Generally, cationic lipids are amphiphilic substances containing a positive hydrophilic head group, two (or more) lipophilic tails or steroid moieties, and a linker between these two domains.Preferably, the cationic lipid carries a net positive charge at approximately physiological pH.Cationic liposomes have traditionally been the most commonly used non-viral delivery system for oligonucleotides, such as plasmid DNA, antisense oligos, and siRNA / small hairpin RNA-shRNA. Cationic lipids, such as DOTAP (1,2-dioleoyl-3-trimethylammonium-propane) and DOTMA (N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethyl-ammonium methylsulfate), can form complexes or lipoplexes with negatively charged nucleic acids through electrostatic interactions, providing high in vitro transfection efficiency.

[0359] In the compositions of the present disclosure, the cationic lipid may be, for example, N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), 1,2-dioleoyltrimethylammonium propane chloride (DOTAP) (also known as N-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride and 1,2-dioleyloxy-3-trimethylaminopropane chloride salt), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride, and 1,2-dioleyloxy-3-trimethylaminopropane chloride salt. (Dimethyl)-N,N,N-trimethylammonium chloride (DOTMA), N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), 1,2-di-y-linolenyloxy-N,N-dimethylaminopropane (γ-DLenDMA), 1,2-dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-dilinoleyl 1,2-Dilinoleyloxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-Dilinoleyloxy-3-morpholinopropane (DLin-MA), 1,2-Dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-Dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1-Linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-Dilinoleyloxy-3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2 -Dilinoleoyl-3-trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), or 3-(N,N-dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-dioleylamino)-1,2-propanediol (DOAP), 1,2-dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA) or their analogs, (3aR,5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9,12-dienyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxol-5-amine, (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraene-19 -yl 4-(dimethylamino)butanoate (MC3), 1,1'-(2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazin-1-yl)ethylazanediyl)didodecan-2-ol (C12-200), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-diol The compound may be 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (DLin-M-C3-DMA), 3-((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yloxy)-N,N-dimethylpropan-1-amine (MC3 ether), 4-((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yloxy)-N,N-dimethylbutan-1-amine (MC4 ether), or any combination thereof. Other cationic lipids include N,N-distearyl-N,N-dimethylammonium bromide (DDAB), 3P-(N-(N',N'-dimethylaminoethane)-carbamoyl)cholesterol (DC-Choi), N-(1-(2,3-dioleyloxy)propyl)-N-2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoroacetate (DOSPA), dioctadecylamidoglycylcarboxyspermine (DOGS), 1,2-dioleoyl-sn-3-phosphoethanolamine (DOPE), 1,2-dioleoyl-3-dimethylammonium propane (DODAP), N-(1,2-dimyristyloxyprop-3-yl)-N,Examples of suitable cationic lipids include, but are not limited to, N-dimethyl-N-hydroxyethylammonium bromide (DMRIE) and 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (XTC). Additionally, commercially available preparations of cationic lipids can be used, such as, for example, Lipofectin (including DOTMA and DOPE, available from GIBCO / BRL) and Lipofectamine (including DOSPA and DOPE, available from GIBCO / BRL).

[0360] Other suitable cationic lipids are disclosed in International Publication Nos. WO09 / 086558, WO09 / 127060, WO10 / 048536, WO10 / 054406, WO10 / 088537, WO10 / 129709, and WO2011 / 153493; U.S. Patent Publication Nos. 2011 / 0256175, 2012 / 0128760, and 2012 / 0027803; U.S. Patent No. 8,158,601; and Love et al., PNAS, 107(5), 1864-69, 2010, the contents of which are incorporated herein by reference.

[0361] Other suitable cationic lipids include those with alternative fatty acid groups and other dialkylamino groups, such as those with different alkyl substituents (e.g., N-ethyl-N-methylamino- and N-propyl-N-ethylamino-). These lipids are part of a subcategory of cationic lipids called amino lipids. In some embodiments of the lipid formulations described herein, the cationic lipid is an amino lipid. In general, amino lipids with less saturated acyl chains are easier to size, especially when the complexes must be sized to about 0.3 microns or less for filter sterilization. Carbon chain lengths of C 14 ~C 22 Other scaffolds can be used to separate the amino group and the fatty acid or fatty alkyl portion of the amino lipid.

[0362] In some embodiments, the compositions described herein comprise a cationic lipid of Formula I according to patent application PCT / EP2017 / 064066, the disclosure of which is also incorporated herein by reference in this context.

[0363] In some embodiments, the amino lipids or cationic lipids of the present disclosure are ionizable and have at least one protonatable or deprotonatable group. As a result, the lipids have a positive charge at a pH below physiological pH (e.g., pH 7.4) and are neutral at a second pH (preferably above physiological pH). Of course, it is understood that the addition or removal of protons as a function of pH is an equilibrium process, and reference to charged or neutral lipids refers to the nature of the predominant species and does not require that all lipids exist in a charged or neutral form. Lipids having two or more protonatable or deprotonatable groups or that are zwitterionic are not excluded from use in the present disclosure. In certain embodiments, the protonatable lipids have a pKa of the protonatable group in the range of about 4 to about 11. In some embodiments, the ionizable cationic lipids have a pKa of about 5 to about 7. In some embodiments, the pKa of the ionizable cationic lipids is about 6 to about 7.

[0364] Cationic lipid compounds can be combined with drugs to form microparticles, nanoparticles, liposomes or micelles.The drugs delivered by particles, liposomes or micelles can be in gas, liquid or solid form, and the drugs can be polynucleotides, proteins, peptides or small molecules.These particles can then be combined with pharmaceutical excipients as needed to form pharmaceutical compositions.

[0365] This description provides novel cationic lipid compounds and drug delivery systems based on the use of such cationic lipid compounds. The systems can be used in pharmaceutical / drug delivery technologies to deliver polynucleotides, proteins, small molecules, peptides, antigens, or drugs to patients, tissues, organs, or cells. These novel compounds can also be used as coatings, additives, excipients, materials, or materials for biotechnology.

[0366] The cationic lipid compounds described herein offer several different applications in drug delivery technology. The amine-containing moieties of cationic lipid compounds can be used to complex polynucleotides, thereby enhancing their delivery and preventing their degradation. Cationic lipid compounds can also be used to form picoparticles, nanoparticles, microparticles, liposomes, and micelles containing the drug to be delivered. Preferably, the cationic lipid compounds are biocompatible and biodegradable, and the particles formed are also biodegradable and biocompatible and can be used to provide controlled, sustained release of the drug to be delivered. These and their corresponding particles can also respond to pH changes, given that they are protonated at lower pH levels. They can also function as proton sponges in the delivery of drugs to cells, causing endosomolysis.

[0367] In certain embodiments, the cationic lipid compounds are relatively non-toxic. They can be biocompatible and biodegradable. The cationic lipids can have a measured pKa (in a formulation environment) ranging from about 5.5 to about 7.5, more preferably from about 6.0 to about 7.0. They can be designed to have a desired pKa of about 3.0 to about 9.0 or about 5.0 to about 8.0. The cationic lipid compounds described herein are particularly attractive for drug delivery for several reasons: they contain amino groups to interact with DNA, RNA, other polynucleotides, and other negatively charged drugs; to buffer pH; to protect the delivered drug from internal osmotic degradation; they can be synthesized from commercially available starting materials; and / or they are pH-responsive and can be engineered to have a desired pKa.

[0368] Neutral helper lipids Non-limiting examples of non-cationic lipids include lecithin, dialkyloxypropyl (DAA), diacylglycerol (DAG), dimyristoylglycerol (DMG), dioleoylglycerol (DOG), dipalmitoylglycerol (DPG), phosphatidylethanolamine (PE), distearoylglycerol (DSG), lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg yolk sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebrosides, dicetylphosphate, Distearoylglycerophosphocholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoyl-phosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), palmitoyloleoyl-phosphatidylglycerol (POPG), dioleoylphosphatidylethanolamine Examples of suitable phospholipids include 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl-phosphatidylethanolamine (DPPE), dimyristoyl-phosphatidylethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), monomethyl-phosphatidylethanolamine, dimethyl-phosphatidylethanolamine, dielaidoyl-phosphatidylethanolamine (DEPE), stearoyloleoyl-phosphatidylethanolamine (SOPE), lysophosphatidylcholine, dilinoleoylphosphatidylcholine, and mixtures thereof. Other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids can also be used. The acyl group in these lipids is preferably C 10 ~C 24 It is an acyl group derived from a fatty acid having a carbon chain, for example, lauroyl, myristoyl, palmitoyl, stearoyl, or oleyl.

[0369] Further examples of non-cationic lipids include sterols such as cholesterol and its derivatives.Non-limiting examples of cholesterol derivatives include polar analogs such as 5α-cholestanol, 5α-coprostanol, cholesteryl-(2'-hydroxy)-ethyl ether, cholesteryl-(4'-hydroxy)-butyl ether, and 6-ketocholestanol; non-polar analogs such as 5α-cholestan, cholestenone, 5α-cholestanone, cholesteryl decanoate; and mixtures thereof.In a preferred embodiment, the cholesterol derivative is a polar analog such as cholesteryl-(4'-hydroxy)-butyl ether.

[0370] In some embodiments, the non-cationic lipid present in lipid particles comprises or consists of the mixture of one or more phospholipids and cholesterol or its derivatives.In other embodiments, the non-cationic lipid present in lipid particles comprises or consists of one or more phospholipids, for example, cholesterol-free lipid particle formulations.In still other embodiments, the non-cationic lipid present in lipid particles comprises or consists of cholesterol or its derivatives, for example, phospholipid-free lipid particle formulations.

[0371] Other examples of non-cationic lipids include phosphorus-free containing lipids such as, for example, stearylamine, dodecylamine, hexadecylamine, acetyl palmitate, glycerol ricinoleate, hexadecyl stearate, isopropyl myristate, amphoteric acrylic polymers, triethanolamine lauryl sulfate, alkyl-aryl sulfate polyethyloxylated fatty acid amides, dioctadecyldimethylammonium bromide, ceramides, and sphingomyelin.

[0372] In some embodiments, the non-cationic lipid comprises 10 mol% to 60 mol%, 20 mol% to 55 mol%, 20 mol% to 45 mol%, 20 mol% to 40 mol%, 25 mol% to 50 mol%, 25 mol% to 45 mol%, 30 mol% to 50 mol%, 30 mol% to 45 mol%, 30 mol% to 40 mol%, 35 mol% to 45 mol%, 37 mol% to 42 mol%, or 35 mol%, 36 mol%, 37 mol%, 38 mol%, 39 mol%, 40 mol%, 41 mol%, 42 mol%, 43 mol%, 44 mol%, or 45 mol% (or any percentage or range thereof) of the total lipid present in the particle.

[0373] In embodiments in which the lipid particle contains a mixture of phospholipids and cholesterol or a cholesterol derivative, the mixture may comprise up to 40 mol%, 45 mol%, 50 mol%, 55 mol%, or 60 mol% of the total lipid present in the particle.

[0374] In some embodiments, the phospholipid component in the mixture can comprise 2 mol% to 20 mol%, 2 mol% to 15 mol%, 2 mol% to 12 mol%, 4 mol% to 15 mol%, or 4 mol% to 10 mol% (or any percentage or range thereof) of the total lipid present in the particle. In certain preferred embodiments, the phospholipid component in the mixture comprises 5 mol% to 10 mol%, 5 mol% to 9 mol%, 5 mol% to 8 mol%, 6 mol% to 9 mol%, 6 mol% to 8 mol%, or 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, or 10 mol% (or any percentage or range thereof) of the total lipid present in the particle.

[0375] In other embodiments, the cholesterol component in the mixture may comprise 25 mol% to 45 mol%, 25 mol% to 40 mol%, 30 mol% to 45 mol%, 30 mol% to 40 mol%, 27 mol% to 37 mol%, 25 mol% to 30 mol%, or 35 mol% to 40 mol% (or any percentage or range thereof) of the total lipid present in the particle. In certain preferred embodiments, the cholesterol component in the mixture comprises 25 mol% to 35 mol%, 27 mol% to 35 mol%, 29 mol% to 35 mol%, 30 mol% to 35 mol%, 30 mol% to 34 mol%, 31 mol% to 33 mol%, or 30 mol%, 31 mol%, 32 mol%, 33 mol%, 34 mol%, or 35 mol% (or any percentage or range thereof) of the total lipid present in the particle.

[0376] In embodiments in which the lipid particle does not contain phospholipids, cholesterol or a derivative thereof may comprise up to 25 mol%, 30 mol%, 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol%, or 60 mol% of the total lipid present in the particle.

[0377] In some embodiments, cholesterol or a derivative thereof in a phospholipid-free lipid particle formulation may comprise 25 mol% to 45 mol%, 25 mol% to 40 mol%, 30 mol% to 45 mol%, 30 mol% to 40 mol%, 31 mol% to 39 mol%, 32 mol% to 38 mol%, 33 mol% to 37 mol%, 35 mol% to 45 mol%, 30 mol% to 35 mol%, 35 mol% to 40 mol%, or 30 mol%, 31 mol%, 32 mol%, 33 mol%, 34 mol%, 35 mol%, 36 mol%, 37 mol%, 38 mol%, 39 mol%, or 40 mol% (or any percentage or range thereof) of the total lipid present in the particle.

[0378] In other embodiments, the non-cationic lipid comprises 5 mol% to 90 mol%, 10 mol% to 85 mol%, 20 mol% to 80 mol%, 10 mol% (e.g., phospholipids only), or 60 mol% (e.g., phospholipids and cholesterol or a derivative thereof) of the total lipid present in the particle (or any percentage or range thereof).

[0379] The percentage of non-cationic lipid present in the lipid particle is a target amount, and the actual amount of non-cationic lipid present in the formulation can vary, for example, by ±5 mol %.

[0380] A composition containing a cationic lipid compound may contain 30-70% cationic lipid compound, 0-60% cholesterol, 0-30% phospholipid, and 1-10% polyethylene glycol (PEG). Preferably, the composition contains 30-40% cationic lipid compound, 40-50% cholesterol, and 10-20% PEG. In another preferred embodiment, the composition contains 50-75% cationic lipid compound, 20-40% cholesterol, 5-10% phospholipid, and 1-10% PEG. The composition may contain 60-70% cationic lipid compound, 25-35% cholesterol, and 5-10% PEG. The composition may contain up to 90% cationic lipid compound and 2-15% helper lipid.

[0381] The formulation may be, for example, a lipid particle formulation containing 8-30% compound, 5-30% helper lipid, and 0-20% cholesterol; 4-25% cationic lipid, 4-25% helper lipid, 2-25% cholesterol, 10-35% cholesterol-PEG, and 5% cholesterol-amine; or 2-30% cationic lipid, 2-30% helper lipid, 1-15% cholesterol, 2-35% cholesterol-PEG, and 1-20% cholesterol-amine; or up to 90% cationic lipid and 2-10% helper lipid, or even 100% cationic lipid.

[0382] lipid complex In addition to cationic, lipid particles described herein can also contain lipid complexes.The complex lipids are useful in preventing particle aggregation.Suitable complex lipids include but are not limited to PEG-lipid complexes, cationic polymer-lipid complexes, and mixtures thereof.

[0383] In preferred embodiments, lipid complex is PEG-lipid.Examples of PEG-lipid include but are not limited to PEG conjugated to dialkyloxypropyl (PEG-DAA), PEG conjugated to diacylglycerol (PEG-DAG), PEG conjugated to phospholipid, for example, phosphatidylethanolamine (PEG-PE), for example, distearoyl-glycero-phosphoethanolamine (PEG-DSPE), for example, dimyristoyl-glycerol (PEG-DMG), for example, dioleoyl-glycerol (PEG-DOG), for example, dipalmitoyl-glycerol (PEG-DPG), for example, distearoyl-glycerol (PEG-DSG), for example, dimyristoyl-glycero-3-phosphoethanolamine (PEG-DMPE), for example, dipalmitoyl-glycero-3-phosphoethanolamine (PEG-DPPE), PEG conjugated to ceramide, PEG conjugated to cholesterol or its derivatives, and mixtures thereof.

[0384] PEG is a linear water-soluble polymer of ethylene PEG repeating units with two terminal hydroxyl groups. PEGs are classified according to their molecular weight and include: monomethoxypolyethyleneglycol (MePEG-OH), monomethoxypolyethyleneglycol-succinate (MePEG-S), monomethoxypolyethyleneglycol-succinimidylsuccinate (MePEG-S-NHS), monomethoxypolyethyleneglycol-amine (MePEG-NH2), monomethoxypolyethyleneglycol-tresylate (MePEG-TRES), monomethoxypolyethyleneglycol-imidazolyl-carbonyl (MePEG-IM), as well as compounds containing terminal hydroxyl groups instead of terminal methoxy groups (e.g., HO-PEG-S, HO-PEG-S-NHS, HO-PEG-NH2).

[0385] The PEG moiety of the PEG-lipid conjugates described herein may comprise an average molecular weight ranging from 550 daltons to 10,000 daltons. In certain cases, the PEG moiety has an average molecular weight of 750 daltons to 5,000 daltons (e.g., 1,000 daltons to 5,000 daltons, 1,500 daltons to 3,000 daltons, 750 daltons to 3,000 daltons, 750 daltons to 2,000 daltons). In preferred embodiments, the PEG moiety has an average molecular weight of 2,000 daltons or 750 daltons.

[0386] In certain cases, PEG can be optionally substituted with alkyl, alkoxy, acyl or aryl group.PEG can be directly conjugated to lipid, or can be linked to lipid via linker moiety.For example, any linker moiety suitable for linking PEG to lipid can be used, including non-ester-containing linker moiety and ester-containing linker moiety.In a preferred embodiment, linker moiety is non-ester-containing linker moiety. Suitable non-ester containing linker moieties include, but are not limited to, amide (-C(O)NH-), amino (-NR-), carbonyl (-C(O)-), carbamate (-NHC(O)O-), urea (-NHC(O)NH-), disulfide (-SS-), ether (-O-), succinyl (-(O)CCH2CH2C(O)-), succinamidyl (-NHC(O)CH2CH2C(O)NH-), ether, disulfide, and combinations thereof (such as linkers containing both carbamate and amide linker moieties). In a preferred embodiment, a carbamate linker is used to attach PEG to the lipid.

[0387] In other embodiments, an ester-containing linker moiety is used to attach PEG to the lipid. Suitable ester-containing linker moieties include, for example, carbonate (—OC(O)O—), succinoyl, phosphate ester (—O—(O)POH-O—), sulfonate ester, —C(O)O—, —O(O)C—, —NH(O)C—, —C(O)NH—, phosphate, C1-C 10 Alkyl-phosphate, C3-C 10 Alkenyl-phosphate, phosphorothioate, C1-C 10 Alkyl-phosphorothioates, C3-C 10 Alkenyl-phosphorothioates, Boranophosphates, C1-C 10 Alkyl-boranophosphate, C3-C 10 Alkenyl-boranophosphate, -C(O)NH-C-C 10 Alkyl-phosphate, -C(O)NH-C3-C 10 Alkenyl phosphate, -C(O)O-C1-C10 Alkyl-phosphate, -C(O)O-C3-C 10 Alkenyl-phosphate, -C(O)NH-C1-C 10 Alkyl-phosphorothioate, -C(O)NH-C3-C 10 Alkenyl-phosphorothioate, -C(O)O-C1-C 10 Alkyl-phosphorothioate, -C(O)O-C3-C 10 Alkenyl-phosphorothioate, -C(O)-NH-C-C 10 Alkyl-boranophosphate, -C(O)-NH-C3-C 10 Alkenyl-boranophosphate, -C(O)O-C1-C 10 Alkyl-boranophosphate or -C(O)O-C3-C 10 alkenyl-boranophosphates, and combinations thereof.

[0388] Phosphatidylethanolamines with various acyl chain groups of varying chain length and saturation can be conjugated to PEG to form lipid conjugates. Such phosphatidylethanolamines are commercially available or can be isolated or synthesized using conventional techniques known to those skilled in the art. 10 ~C 20 Phosphatidylethanolamines containing saturated or unsaturated fatty acids with carbon chain lengths in the range of 0 to 100 are preferred. Phosphatidylethanolamines with mono- or di-unsaturated fatty acids, as well as mixtures of saturated and unsaturated fatty acids, can also be used. Suitable phosphatidylethanolamines include, but are not limited to, dimyristoyl-phosphatidylethanolamine (DMPE), dipalmitoyl-phosphatidylethanolamine (DPPE), dioleoyl-phosphatidylethanolamine (DOPE), and distearoyl-phosphatidylethanolamine (DSPE).

[0389] The term "diacylglycerol" or "DAG" refers to a diacylglycerol consisting of two fatty acyl chains R 1 and R 2and 2-30 carbon atoms, both of which are independently attached to the 1- and 2-positions of glycerol by ester linkages. The acyl groups can be saturated or have different degrees of unsaturation. Suitable acyl groups include lauroyl (C 12 ), myristoyl (C 14 ), palmitoyl (C 16 ), stearoyl (C 18 ), and Icosoil (C 20 In a preferred embodiment, R 1 and R 2 are identical, i.e., R 1 and R 2 are both myristoyl (i.e., dimyristoyl), and R 1 and R 2 are both stearoyl (i.e., distearoyl).

[0390] The term "dialkyloxypropyl" or "DAA" includes compounds with two alkyl chains, both of which, independently, have between 2 and 30 carbons. The alkyl groups can be saturated or have varying degrees of unsaturation.

[0391] Preferably, the PEG-DAA conjugate is PEG-didecyloxypropyl (C 10 ) conjugate, PEG-dilauryloxypropyl (C 12 ) conjugate, PEG-dimyristyloxypropyl (C 14 ) conjugate, PEG-dipalmityloxypropyl (C 16 ) conjugate, or PEG-distearyloxypropyl (C 18 ) conjugates. In these embodiments, the PEG preferably has an average molecular weight of 750 or 2,000 daltons. In certain embodiments, the terminal hydroxyl groups of the PEG are replaced with methyl groups.

[0392] In addition to the above, other hydrophilic polymers can be used in place of PEG. Examples of suitable polymers that can be used in place of PEG include, but are not limited to, polyvinylpyrrolidone, polymethyloxazoline, polyethyloxazoline, polyhydroxypropylmethacrylamide, polymethacrylamide and polydimethylacrylamide, polylactic acid, polyglycolic acid, and derivatized celluloses such as hydroxymethylcellulose or hydroxyethylcellulose.

[0393] In some embodiments, the lipid conjugates (e.g., PEG-lipid) comprise 0.1 mol% to 2 mol%, 0.5 mol% to 2 mol%, 1 mol% to 2 mol%, 0.6 mol% to 1.9 mol%, 0.7 mol% to 1.8 mol%, 0.8 mol% to 1.7 mol%, 0.9 mol% to 1.6 mol%, 0.9 mol% to 1.8 mol%, 1 mol% to 1.8 mol%, 1 mol% to 1.7 mol%, 1.2 mol% to 1.8 mol%, 1.2 mol% to 1.7 mol%, 1.3 mol% to 1.6 mol%, or 1.4 mol% to 1.5 mol% (or any percentage or range thereof) of the total lipid present in the particle. In other embodiments, the lipid conjugate (e.g., PEG-lipid) comprises 0 mol% to 20 mol%, 0.5 mol% to 20 mol%, 2 mol% to 20 mol%, 1.5 mol% to 18 mol%, 2 mol% to 15 mol%, 4 mol% to 15 mol%, 2 mol% to 12 mol%, 5 mol% to 12 mol%, or 2 mol% (or any percentage or range thereof) of the total lipid present in the particle.

[0394] In further embodiments, the lipid conjugate (e.g., PEG-lipid) comprises 4 mol% to 10 mol%, 5 mol% to 10 mol%, 5 mol% to 9 mol%, 5 mol% to 8 mol%, 6 mol% to 9 mol%, 6 mol% to 8 mol%, or 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, or 10 mol% (or any percentage or range thereof) of the total lipid present in the particle.

[0395] The percentage of lipid complexes (e.g., PEG-lipids) present in the lipid particles of the present invention is a target amount, and the actual amount of lipid complexes present in the formulation may vary, for example, by ±2 mol%. Those skilled in the art will understand that the concentration of lipid complexes may vary depending on the rate at which the lipid complexes used and lipid particles become fusogenic.

[0396] By controlling the composition and concentration of the lipid complexes, the rate at which the lipid complexes are exchanged out of the lipid particles, as well as the rate at which the lipid particles become fusogenic, can be controlled.In addition, other variables, including, for example, pH, temperature, or ionic strength, can be used to change and / or control the rate at which the lipid particles become fusogenic.Other methods that can be used to control the rate at which the lipid particles become fusogenic will be apparent to those skilled in the art upon reading this disclosure.In addition, by controlling the composition and concentration of the lipid complexes, the particle size of the lipids can be controlled.

[0397] Compositions and Formulations for Administration The nucleic acid-lipid composition of the present disclosure can be administered by various routes, for example, by intravenous route, parenteral route, intraperitoneal route or local route, to achieve systemic delivery.In some embodiments, siRNA can be delivered intracellularly, for example, into the cells of target tissues such as lung or liver, or into the cells of inflamed tissue.In some embodiments, the present disclosure provides a method for in vivo siRNA delivery.The nucleic acid-lipid composition can be administered intravenously, subcutaneously, or intraperitoneally to a subject.In some embodiments, the present disclosure provides a method for in vivo delivery of interfering RNA to the lung of a mammalian subject.

[0398] In some embodiments, the present disclosure provides a method for treating a disease or disorder in a mammalian subject, wherein a therapeutically effective amount of a composition of the present disclosure containing a nucleic acid, a cationic lipid, an amphiphile, a phospholipid, cholesterol, and PEG-conjugated cholesterol can be administered to a subject having a disease or disorder associated with expression or overexpression of a gene that can be reduced, decreased, downregulated, or silenced by the composition.

[0399] The compositions and methods of the present disclosure can be administered to a subject by a variety of mucosal administration methods, including oral, rectal, vaginal, intranasal, pulmonary, or transdermal or dermal delivery, or by topical delivery to the eye, ear, skin, or other mucosal surface. In some aspects of the present disclosure, the mucosal tissue layer comprises an epithelial cell layer. The epithelial cells can be pulmonary epithelial cells, airway epithelial cells, bronchial epithelial cells, alveolar epithelial cells, nasal epithelial cells, oral epithelial cells, epidermal epithelial cells, or gastrointestinal epithelial cells. The compositions of the present disclosure can be administered using conventional actuators, such as mechanical spray devices, as well as pressurized, electrically, or other types of actuators.

[0400] The compositions of the present disclosure may be administered in the form of an aqueous solution as a nasal or pulmonary spray, or may be dispensed in spray form by various methods known to those skilled in the art. Pulmonary delivery of the compositions of the present disclosure is achieved, for example, by administering the composition in the form of droplets, particles, or spray, which may be aerosolized, atomized, or nebulized. The particles, spray, or aerosol of the composition may be in either liquid or solid form. A suitable system for dispensing a solution as a nasal spray is described in U.S. Pat. No. 4,511,069. Such a formulation can be conveniently prepared by dissolving a composition according to the present disclosure in water to form an aqueous solution and sterilizing the solution. This formulation can be dispensed in a multi-dose container, for example, the sealed dispensing system disclosed in U.S. Pat. No. 4,511,069. Other suitable nasal spray delivery systems are described in Transdermal Systemic Medication, Y.W. Chien, ed., Elsevier Publishers, New York, 1985; and U.S. Pat. No. 4,778,810. Additional aerosol delivery forms may include, for example, compressed air nebulizers, jet nebulizers, ultrasonic nebulizers, and piezoelectric nebulizers, which deliver biologically active agents dissolved or suspended in a pharmaceutical solvent, such as water, ethanol, or mixtures thereof.

[0401] The nasal and pulmonary spray solutions of the present disclosure typically contain a drug or drugs for delivery formulated with, optionally, a surfactant, e.g., a non-ionic surfactant (e.g., polysorbate-80), and one or more buffers. In some embodiments of the present disclosure, the nasal spray solution further comprises a propellant. The pH of the nasal spray solution may be approximately pH 6.8 to 7.2. Alternatively, the drug solvent used may be a weakly acidic aqueous buffer with a pH of 4 to 6. Other ingredients may be added to improve or maintain chemical stability, including preservatives, surfactants, dispersants, or gases.

[0402] In some embodiments, a pharmaceutical product is disclosed that includes a solution containing a composition of the present disclosure and an actuator for a pulmonary, mucosal, or intranasal spray or aerosol.

[0403] The dosage form of the compositions of the present disclosure may be a liquid in the form of droplets or an emulsion, or in the form of an aerosol.

[0404] The dosage form of the composition of the present disclosure may be a solid, which can be reconstituted in a liquid before administration. This solid may be administered as a powder. This solid may be in the form of a capsule, tablet, or gel.

[0405] To formulate a composition for pulmonary delivery within the scope of the present disclosure, the biologically active substance can be mixed with various pharmaceutically acceptable excipients and bases or carriers for dispersing the active substance(s). Examples of excipients include pH adjusters, such as arginine, sodium hydroxide, glycine, hydrochloric acid, citric acid, and mixtures thereof. Other excipients include local anesthetics (e.g., benzyl alcohol), tonicity agents (e.g., sodium chloride, mannitol, sorbitol), adsorption inhibitors (e.g., Tween 80), solubility enhancers (e.g., cyclodextrins and their derivatives), stabilizers (e.g., serum albumin), and reducing agents (e.g., glutathione). When the composition for mucosal delivery is a liquid, the osmolality of the formulation, measured relative to the osmolality of 0.9% (w / v) saline solution (defined as 1), is typically adjusted to a value that does not cause substantial irreversible tissue damage to the mucosa at the administration site. Generally, the osmolality of the solution is adjusted to a value of about 1 / 3 to 3, more typically 1 / 2 to 2, and most often 3 / 4 to 1.7.

[0406] The biologically active substance may be dispersed in a base or vehicle, which may include a hydrophilic compound capable of dispersing the active substance and any desired excipients. The base can be selected from a variety of suitable carriers, including, but not limited to, polycarboxylic acids or their salts, copolymers of carboxylic acid anhydrides (e.g., maleic anhydride) with other monomers (e.g., methyl (meth)acrylate, acrylic acid, etc.), hydrophilic vinyl polymers such as polyvinyl acetate, polyvinyl alcohol, polyvinylpyrrolidone, cellulose derivatives such as hydroxymethylcellulose, hydroxypropylcellulose, and natural polymers such as chitosan, collagen, sodium alginate, gelatin, hyaluronic acid, and their non-toxic metal salts. Biodegradable polymers, such as polylactic acid, poly(lactic acid-glycolic acid) copolymers, polyhydroxybutyric acid, poly(hydroxybutyric acid-glycolic acid) copolymers, and mixtures thereof, are often selected as the base or carrier. Alternatively or additionally, synthetic fatty acid esters, such as polyglycerin fatty acid esters and sucrose fatty acid esters, can also be used as carriers. Hydrophilic polymers and other carriers can be used alone or in combination, and the structural integrity of the carrier can be improved by partial crystallization, ionic bonding, crosslinking, etc. Carriers can be provided in a variety of forms, including fluid or viscous solutions, gels, pastes, powders, microspheres, and films, and applied directly to the nasal mucosa. In this regard, the use of selected carriers can result in enhanced absorption of the biologically active substance.

[0407] Formulations for mucosal, nasal, or pulmonary delivery may contain hydrophilic low-molecular-weight compounds as bases or excipients. Such hydrophilic low-molecular-weight compounds provide a medium through which water-soluble active substances, such as biologically active peptides or proteins, can diffuse through the base to the body surface, where they are absorbed. The hydrophilic low-molecular-weight compounds may absorb moisture from the mucous membrane or administration environment, dissolving the water-soluble active peptides. The molecular weight of hydrophilic low-molecular-weight compounds is generally 10,000 or less, preferably 3000 or less. Examples of hydrophilic low-molecular-weight compounds include polyol compounds, such as oligosaccharides, disaccharides, and monosaccharides, including sucrose, mannitol, lactose, L-arabinose, D-erythrose, D-ribose, D-xylose, D-mannose, D-galactose, lactulose, cellobiose, gentiose, glycerin, polyethylene glycol, and mixtures thereof. Further examples of hydrophilic low molecular weight compounds include N-methylpyrrolidone, alcohols (eg, oligovinyl alcohol, ethanol, ethylene glycol, propylene glycol, etc.), and mixtures thereof.

[0408] Alternatively, the compositions of the present disclosure may contain, as pharmaceutically acceptable carrier materials, substances required to approximate physiological conditions, such as pH adjusting and buffering agents, osmotic pressure adjusting agents, and wetting agents, such as sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, and mixtures thereof. In the case of solid compositions, conventional non-toxic pharmaceutically acceptable carriers may be used, including, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, magnesium carbonate, and the like.

[0409] In certain embodiments of the present disclosure, the biologically active agent may be administered as a sustained-release formulation, for example, as a composition containing a sustained-release polymer. The active agent may be prepared using a carrier that prevents rapid release, for example, a controlled-release vehicle, such as a polymer, a microencapsulated delivery system, or a bioadhesive gel. Sustained delivery of the active agent in various compositions of the present disclosure can be achieved by including in the composition an agent that delays absorption, for example, aluminum monostearate hydrogel and gelatin.

[0410] While the present disclosure has been described in connection with particular embodiments and numerous details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that the present disclosure includes further embodiments and that some of the details described herein can be varied considerably without departing from the present disclosure. The present disclosure includes such further embodiments, modifications, and equivalents. In particular, the present disclosure includes any combination of features, expressions, or elements of the various exemplary components and examples. The present disclosure includes the following embodiments. [Section 1] Compounds of Formula IA [ka] or a pharmaceutically acceptable salt or solvate thereof, wherein: X 1 、X 2 and X 3 are each independently, C 1 -C 10 Alkyl, -(CH 2 ) m -O-(CH 2 ) n - and -(CH 2 ) m -NR N -(CH 2 ) n wherein n is 1 to 36, m is 1 to 30, and R N is H, methyl, (-CH 2 F), (CHF 2 ), or (-CF 3 ) and; Y 1 、Y 2 and Y 3 are each independently -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, and P(Z)(OH)O 2 wherein Z is O or S; L 1 、L 2 and L 3 are each independently, C 1 -C 10 Alkyl, -(CH 2 ) e -O-(CH 2 ) f -, -(CH 2 ) e -S-(CH 2 ) f -, -(CH 2 ) e -S(O) 2 -(CH 2 ) f -, -(CH 2 ) e -NR N -(CH 2 ) f - and -(CH 2 -CH 2 -O) k (CH 2 ) 2 wherein e is 1 to 10, f is 1 to 16, k is 1 to 20, and R N is H, methyl, or CH 2 F, CHF 2 , or CF 3 and; G 1 、G 2 and G 3 are each independently selected from the group consisting of monosaccharides, monosaccharide derivatives, vitamins, polyols, polysialic acids, and polysialic acid derivatives; X 4 teeth, (a)-(CH 2 )g -O-(CH 2 ) h -or-(CH 2 ) g -NR N -(CH 2 ) h -, wherein g is 1 to 30, h is 1 to 36, and R N is H, methyl, or CH 2 F, CHF 2 , or CF 3 and (b) an amino acid, and (c)-NHC(O)R 2 , where R 2 is C 1 -C 10 Alkyl, carbocycle, heterocyclyl, heteroaryl, C 1 -C 10 Alkyl-Carbocyclic, C 1 -C 10 Alkyl-heterocyclyl or C 1 -C 10 alkyl-heteroaryl, and wherein R 2 is optionally substituted; is selected from the group consisting of Q is absent, alkylamino, -C(O)-(CH 2 ) i -, -(CH 2 ) i -O-(CH 2 ) j -, -(CH 2 ) i -NR 3 -(CH 2 ) j -, -(CH 2 ) i -SS-(CH 2 ) j -, -(CH 2 ) i -S-(CH 2 ) j -, -(CH 2 ) i -S(O) 2 -(CH 2 ) j -, -(CH 2 ) i -NHC(O)-(CH 2 ) j -, -(CH 2 ) i -C(O)NH-(CH 2 ) j -, -(CH 2 ) i -SC(O)-(CH 2 ) j - or -(CH 2 ) i -C(O)S-(CH 2 ) j -, wherein i is 1 to 30; j is 1 to 36; and R 3 is hydrogen or alkyl; L 4 is absent, -C(O)O-, -C(O)NH-, phosphate, C 1 -C 10 Alkyl-phosphate, C 3 -C 10 Alkenyl-phosphate, phosphorothioate, C 1 -C 10 Alkyl-phosphorothioate, C 3 -C 10 Alkenyl-phosphorothioate, Boranophosphate, C 1 -C 10 Alkyl-boranophosphate, C 3 -C 10 Alkenyl-boranophosphate, -C(O)NH-C 1 -C 10 Alkyl-phosphate, -C(O)NH-C 3 -C 10 Alkenyl-phosphate, -C(O)OC 1 -C 10 Alkyl-phosphate, -C(O)OC 3 -C 10 Alkenyl-phosphate, -C(O)NH-C 1 -C 10 Alkyl-phosphorothioate, -C(O)NH-C 3 -C 10 Alkenyl-phosphorothioate, -C(O)OC 1 -C 10 Alkyl-phosphorothioate, -C(O)OC 3 -C10 Alkenyl-phosphorothioate, -C(O)-NH-C 1 -C 10 Alkyl-boranophosphate, -C(O)-NH-C 3 -C 10 Alkenyl-boranophosphate, -C(O)OC 1 -C 10 Alkyl-boranophosphate or -C(O)OC 3 -C 10 alkenyl-boranophosphate; and R 1 is a biologically active molecule, or a pharmaceutically acceptable salt or solvate thereof. [Section 2] X 1 、X 2 and X 3 are each independently (-CH 2 ) m -O-CH 2 -, wherein m is 1 to 4. [Section 3] X 1 、X 2 and X 3 are each independently (-CH 2 ) 2 -O-CH 2 Item 2. The compound according to item 1, wherein [Section 4] Y 1 、Y 2 and Y 3 and each is -NHC(O)- or -C(O)NH-. [Section 5] Y 1 、Y 2 and Y 3 Each of the groups represented by the formula (I) is -NHC(O)-. [Section 6] L 1 、L 2 and L 3 are each independently, C 3 -C 8 Alkyl or -(CH 2 -CH 2 -O) k (CH 2 ) 2 -, wherein k is 1 to 10. [Section 7] L 1 、L 2 and L 3 are each independently -(CH 2 -CH 2 -O) k (CH 2 ) 2 -, wherein k is 2 to 4. [Section 8] L 1 、L 2 and L 3 are respectively, C 1 -C 10 Item 6. The compound according to any one of items 1 to 5, wherein the aryl group is alkyl. [Section 9] G 1 、G 2 and G 3 each independently represents folic acid, ribose, retinol, niacin, riboflavin, biotin, glucose, mannose, fucose, sucrose, lactose, mannose-6-phosphate, N-acetylgalactosamine, N-acetylglucosamine, sialic acid, sialic acid derivatives, allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, fucitol, fucosamine, fucose, fuculose, galactosamine, galactosaminitol, galactose, glucosamine, glucosaminitol, glucose-6 Item 9. The compound according to any one of Items 1 to 8, wherein the compound is selected from the group consisting of phosphate, guloseglyceraldehyde, glycero-mannoseptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, psicose, quinovose, quinovosamine, rhamnitol, rhamnosamine, rhamnose, ribulose, sedoheptulose, sorbose, tagatose, talose, threose, xylose, and xylulose. [Section 10] G 1 、G 2 and G 3 and each represents N-acetylgalactosamine. [Section 11] X 4 but,

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[0411] The present disclosure is further illustrated in the following examples, which do not limit the scope of the present disclosure herein.

[0412] Example 1: Synthesis of Complex 1 An exemplary oligonucleotide conjugate 1 was synthesized using compound 7, shown below, and prepared as shown below in Scheme 1. [ka] wherein, for purposes of Example 1, the substituent [ka] represents mFVII ASO (murine Factor VII antisense oligonucleotide) connected to the 5' end of the mFVII ASO via a hexylamino linker. Scheme 1 [ka]

[0413] Complex 1: Steps 1 and 2 To an equimolar mixture of 5.05 g (10 mmol) of 1-(di-tert-butyl 3,3'-((2-amino-2-((3-(tert-butoxy)-3-oxopropoxy)methyl)propane-1,3-diyl)bis(oxy))dipropionate (prepared according to J. Org. Chem. 2002, 67, 1411-1413) and 2.63 g (10 mmol) of 1-((benzyloxy)carbonyl)piperidine-4-carboxylic acid was added 3.80 g of HATU (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide). A mixture of 3.72 g (5 mL, 30 mmol) of DIEA (diisopropylethylamine) and 3.72 g (5 mL, 30 mmol) of DIEA (diisopropylethylamine) was stirred in 50 mL of anhydrous DMF (N,N-dimethylformamide) at room temperature for 12-16 hours. The mixture was diluted with ethyl acetate (200 mL), washed with water (300 mL), washed with saline (200 mL), dried over NaSO, and the solvent was evaporated under reduced pressure. The residue was purified by filtration on a silica gel column (80 g, Teledyne) on a Combiflash® system with an ethyl acetate:hexane gradient (0-100% over 30 minutes). The product was purified by ISCO. The product was eluted with 30–35% ethyl acetate:hexane fractions. The product-containing fractions were pooled, and the solvent was evaporated under reduced pressure to give 6 g (80%) of the product as a colorless syrup. Mass: 750.43 (calculated), 773.60 (M+Na, observed). Then, to a solution of 5.6 g of the product from Step 1 in methanol (60 mL) was added 560 mg of 10% wt / wt Pd—C (10%, wet Degussa type). The mixture was hydrogenated under a hydrogen-filled balloon for 18 h. The mixture was filtered through a pad of Celite®, and the Celite® pad was washed with 30 mL of methanol. The filtered methanol solution was evaporated to give 4.5 g of 2 as a colorless solid. Mass: 616.8 (calculated), 617.8 (M+H, observed). [ka]

[0414] Complex 1: Step 3 To a solution of 4.5 g (7.3 mmol) of 2 in 50 mL of anhydrous DMF, monobenzylpentanedioic acid (1.62 g, 7.3 mmol) was added, followed by DIEA (2.4 g, 19.2 mmol) and HATU (2.8 g, 7.3 mmol). The mixture was stirred at room temperature overnight. The mixture was then diluted with ethyl acetate (120 mL), washed with water (2 × 150 mL), washed with brine (1 × 150 mL), dried over NaSO, and the solvent was evaporated under reduced pressure. The residue was purified on a Teledyne ISCO silica gel column (80 g) using a methanol:dichloromethane gradient. Product-containing fractions were eluted with 12–15% methanol:dichloromethane and combined. The solvent was then evaporated under reduced pressure to give 4.75 g of 3. Mass: 821.02 (calculated), 843.4 (M + Na, observed). [ka]

[0415] Complex 1: Steps 4 and 5 To a solution of 4.7 g of 3 in 45 mL of dichloromethane, 45 mL of TFA (trifluoroacetic acid) was added, and the mixture was stirred at room temperature for 16 h. The solvent was evaporated under reduced pressure and placed under high vacuum for 24 h to give a quantitative yield of the product (3.7 g). Subsequently, to a solution of 3.1 g (4.7 mmol) of the product isolated from Step 4 in 40 mL of anhydrous DMF, DIEA (4.7 g, 38 mmol) was added, followed by the slow addition of pentafluorophenyl trifluoroacetate (5.3 g, 18.8 mmol). The mixture was stirred at room temperature for 16 h. The mixture was diluted with 250 mL of ethyl acetate and washed with 250 mL of saturated aqueous sodium bicarbonate. The aqueous layer was washed with 100 mL of ethyl acetate. The combined organic solution was washed with water (200 mL), brine (200 mL), dried over Na2SO4, filtered, and the solvent was evaporated. The residue was purified on an 80 g Teledyne ISCO silica gel column using an ethyl acetate:hexane gradient. The product was eluted with 60-70% ethyl acetate:hexane fractions and monitored by TLC (thin layer chromatography). Pure fractions were combined and the solvent evaporated under reduced pressure to give 1.9 g of 4. Mass: 1150.84 (calculated), 1173.2 (M-H+Na, observed). [ka]

[0416] Complex 1: Step 6 To a solution of 3.1 g (2.69 mmol) of 4 and 4.9 g (8.44 mmol, 3.1 equiv.) of 5 (see literature procedures presented in International Journal of Peptide & Protein Research, Volume 43 Issue 5 Pages 477-85, 1994; Bioorganic & Medicinal Chemistry, 13(10), 3553-3564; 2005; US 20140031533 A1; and WO 2015 / 168514 A1) in 1:1 v / v ethyl acetate:acetonitrile (60 mL), palladium hydroxide on charcoal (10-20% loading, 1 g) was added, and the mixture was hydrogenated under a hydrogen-filled balloon for 18 h. The mixture was filtered through a pad of Celite®, and the Celite® pad was washed with 40 mL of acetonitrile. The solvent was then evaporated, and the residue was purified on a Teledyne ISCO silica gel column (80 g) using a methanol / dichloromethane gradient. The major product was eluted with 20-25% methanol / dichloromethane. Fractions containing pure product were then combined, and the solvent was evaporated under reduced pressure to give 1.4 g of pure 6. Mass: 1848.02 (calculated), 1870.1 (M-H+Na, observed). [ka]

[0417] Complex 1: Step 8 To a solution of 6 (360 mg, 0.19 mmol) in anhydrous N,N-dimethylformamide (4 mL) in a 50 mL round-bottom flask under an argon atmosphere, triethylamine (57 mg, 0.57 mmol) was added, followed by pentafluorophenyl trifluoroacetate (106 mg, 0.38 mmol), and the reaction mixture was stirred at room temperature for 2.5 h. The solution was cooled in an ice bath and quenched with saturated aqueous sodium bicarbonate solution (1.5 mL). The resulting solution was diluted with water (2 mL) and extracted with ethyl acetate (2 × 3 mL). The combined organic solution was washed with 1N aqueous sodium bisulfate, dried (anhydrous sodium sulfate), filtered, and concentrated under reduced pressure. The residue was purified on a Teledyne ISCO Gold silica gel column (12 g) using a methanol:dichloromethane gradient. The major product was eluted with 12–14% methanol:dichloromethane. Fractions containing the major product were combined, and the solvent was evaporated. The residue was dried under vacuum in the presence of phosphorus pentoxide to give 300 mg of pure 7. Mass: 2012.87 (calculated), 2035.7 (M+Na, observed). [ka]

[0418] Complex 1: Step 8 Compound 7 was then used in post-synthesis conjugation to the 5' end of a mouse Factor VII antisense oligonucleotide via a hexylamino linker using established protocols for oligonucleotide synthesis, yielding conjugate 1. Thus, the oligonucleotide was prepared with an amino group at the 5' end using standard phosphoramidite chemistry. After purification by IE-HPLC (ion-exchange high-performance liquid chromatography), the oligonucleotide was suspended in phosphate buffer (pH 7.4) and a solution of ARCT-GalNAc PFP ester (a third-party reagent) in DMSO (dimethyl sulfoxide) was added to the suspension. The reaction was monitored by MALDI-TOF MS (matrix-assisted laser desorption / ionization time-of-flight mass spectrometry). After 2 h, the reaction was complete, and the reaction mixture was lyophilized. The residue was then resuspended in concentrated ammonium hydroxide for 2 h and lyophilized. The residue was purified by RP-HPLC (reverse-phase high-performance liquid chromatography) to yield conjugate 1.

[0419] Example 2: Synthesis of Complex 2 Conjugate 2, shown below, was synthesized in a similar manner to Scheme 1 and Step 7 of Conjugate 1 using 13, prepared as shown below in Scheme 2. [ka] wherein, for purposes of Example 2, the substituent [ka] represents mFVII ASO (murine Factor VII antisense oligonucleotide) connected to the 5' end of the mFVII ASO via a hexylamino linker. Scheme 2 [ka]

[0420] Scheme 2: Reagents and conditions: i) HATU, DIEA, DMF, room temperature, overnight; ii) Pd-C (10%), 10% wt / wt, H (balloon), MeOH, room temperature, overnight; iii) dipentanoic acid mono-benzyl ester, HATU, DIEA, DMF, room temperature, overnight; iv) TFA:DCM (dichloromethane) (1:1 v / v), room temperature, 16 hours; v) pentafluorophenyl trifluoroacetate, DIEA, room temperature, overnight; vi) 5, Pd(OH)-C (20%), 20% (wt / wt), H (balloon), ethyl acetate:acetonitrile (1:1 v / v), overnight; vii) pentafluorophenyl trifluoroacetate, DIEA, room temperature, overnight.

[0421] Example 3: Synthesis of Complex 3 Conjugate 3, shown below, was synthesized in a similar manner to Scheme 1 for Conjugate 1 using compound 19, prepared as shown below in Scheme 3. [ka] wherein, for purposes of Example 3, the substituent [ka] represents mFVII ASO (murine Factor VII antisense oligonucleotide) connected to the 5' end of the mFVII ASO via a hexylamino linker. Scheme 3 [ka]

[0422] Scheme 3: Reagents and conditions: i) HATU, DIEA, DMF, room temperature, overnight; ii) Pd-C (10%), 10% wt / wt, H (balloon), MeOH, room temperature, overnight; iii) dipentanoic acid mono-benzyl ester, HATU, DIEA, DMF, room temperature, overnight; iv) TFA:DCM (1:1 v / v), room temperature, 16 h; v) pentafluorophenyl trifluoroace...

Claims

1. Compound of Formula IA 【Chemistry 1】 or a pharma- ceutically acceptable salt or solvate thereof, wherein: X １ , X ２ and X ３ are each independently １ -C １０ Alkyl, -(CH ２ ) ｍ -O-(CH ２ ) ｎ - and - (CH ２ ) ｍ -NR Ｎ - (CH ２ ) ｎ -, wherein n is 1 to 36, m is 1 to 30, and R Ｎ is H, methyl, (-CH ２ F), (CHF ２ ), or (-CF ３ ) and Y １ , Y ２ and Y ３ each independently represents -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, or P(Z)(OH)O ２ wherein Z is O or S; L １ , L ２ and L ３ are each independently １ -C １０ Alkyl, -(CH ２ ) ｅ -O-(CH ２ ) ｆ -, -(CH ２ ) ｅ -S-(CH ２ ) ｆ -, -(CH ２ ) ｅ -S (O) ２ - (CH ２ ) ｆ -, -(CH ２ ) ｅ -NR Ｎ - (CH ２ ) ｆ - and - (CH ２ -CH ２ -O) ｋ (CH ２ ) ２ -, wherein e is 1 to 10, f is 1 to 16, k is 1 to 20, and R Ｎ is H, methyl, or CH ２ F, C.H.F. ２ , or C.F. ３ and G １ , G ２ and G ３ are each independently selected from the group consisting of monosaccharides and monosaccharide derivatives; X ４ is -NHC(O)R ２ and R ２ is carbocycle, heterocyclyl or heteroaryl, and R ２ is optionally substituted; Q is absent, alkylamino, -C(O)-(CH ２ ) ｉ -, -(CH ２ ) ｉ -O-(CH ２ ) ｊ -, -(CH ２ ) ｉ -NR ３ - (CH ２ ) ｊ -, -(CH ２ ) ｉ -S-S-(CH ２ ) ｊ -, -(CH ２ ) ｉ -S-(CH ２ ) ｊ -, -(CH ２ ) ｉ -S (O) ２ - (CH ２ ) ｊ -, -(CH ２ ) ｉ -NHC(O)-(CH ２ ) ｊ -, -(CH ２ ) ｉ -C(O)NH-(CH ２ ) ｊ -, -(CH ２ ) ｉ -SC(O)-(CH ２ ) ｊ - or - (CH ２ ) ｉ -C(O)S-(CH ２ ) ｊ -, where i is 1 to 30; j is 1 to 36; and R ３ is hydrogen or alkyl; L ４ is absent, -C(O)O-, -C(O)NH-, phosphate, C １ -C １０ Alkyl-phosphate, C ３ -C １０ Alkenyl-phosphate, phosphorothioate, C １ -C １０ Alkyl-phosphorothioate, C ３ -C １０ Alkenyl-phosphorothioate, boranophosphate, C １ -C １０ Alkyl-boranophosphate, C ３ -C １０ Alkenyl-boranophosphate, —C(O)NH—C １ -C １０ Alkyl-phosphate, -C(O)NH-C ３ -C １０ Alkenyl-phosphate, —C(O)O—C １ -C １０ Alkyl-phosphate, -C(O)O-C ３ -C １０ Alkenyl-phosphate, -C(O)NH-C １ -C １０ Alkyl-phosphorothioates, -C(O)NH-C ３ -C １０ Alkenyl-phosphorothioate, -C(O)O-C １ -C １０ Alkyl-phosphorothioate, -C(O)O-C ３ -C １０ Alkenyl-phosphorothioate, -C(O)-NH-C １ -C １０ Alkyl-boranophosphate, —C(O)—NH—C ３ -C １０ Alkenyl-boranophosphate, —C(O)O—C １ -C １０ Alkyl-boranophosphate or -C(O)O-C ３ -C １０ alkenyl-boranophosphate; and R １ is a biologically active molecule, or a pharma- ceutically acceptable salt or solvate thereof.

2. X １ , X ２ and X ３ Each independently represents (-CH ２ ) ｍ -O-CH ２ -, wherein m is 1 to 4, or a pharma- ceutically acceptable salt or solvate thereof.

3. Y １ , Y ２ and Y ３ The compound of claim 1 or 2, or a pharma- ceutically acceptable salt or solvate thereof, wherein each of

4. Y １ , Y ２ and Y ３ The compound of claim 1 or 2, or a pharma- ceutically acceptable salt or solvate thereof, wherein each is --NHC(O)--.

5. L １ , L ２ and L ３ Each independently represents -(CH ２ -CH ２ -O) ｋ (CH ２ ) ２ -, wherein k is 2 to 4, or a pharma- ceutically acceptable salt or solvate thereof.

6. L １ , L ２ and L ３ are respectively C １ -C １０ The compound of any one of claims 1 to 4, or a pharma- ceutically acceptable salt or solvate thereof, wherein: R is 0 or 1;

7. G １ , G ２ and G ３ each independently represents ribose, glucose, mannose, fucose, sucrose, lactose, mannose-6-phosphate, N-acetylgalactosamine, N-acetylglucosamine, allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, fucitol, fucosamine, fucose, fuculose, galactosamine, galactosaminitol, galactose, glucosamine, glucosaminitol, glucose-6 The compound according to any one of claims 1 to 6, or a pharma- ceutically acceptable salt or solvate thereof, which is selected from the group consisting of phosphate, guloseglyceraldehyde, glycero-mannoseptose, glycerol, glycerone, gulose, idose, lyxose, mannosamine, psicose, quinovose, quinovosamine, rhamnitol, rhamnosamine, rhamnose, ribulose, sedoheptulose, sorbose, tagatose, talose, threose, xylose and xylulose.

8. G １ , G ２ and G ３ The compound of any one of claims 1 to 6, or a pharma- ceutically acceptable salt or solvate thereof, wherein each is N-acetylgalactosamine.

9. X ４ but, 【Chemistry 2】 wherein X is selected from the group consisting of ４ 9. The compound of any one of claims 1 to 8, or a pharma- ceutically acceptable salt or solvate thereof, wherein:

10. Q is alkylamino, -C(O)-(CH ２ ) ｉ -, -(CH ２ ) ｉ -O-(CH ２ ) ｊ -, -(CH ２ ) ｉ -NR ３ - (CH ２ ) ｊ -, -(CH ２ ) ｉ -S-S-(CH ２ ) ｊ -, -(CH ２ ) ｉ -S-(CH ２ ) ｊ -, -(CH ２ ) ｉ -S (O) ２ - (CH ２ ) ｊ -, -(CH ２ ) ｉ -NHC(O)-(CH ２ ) ｊ -, -(CH ２ ) ｉ -C(O)NH-(CH ２ ) ｊ -, -(CH ２ ) ｉ -SC(O)-(CH ２ ) ｊ - or - (CH ２ ) ｉ -C(O)S-(CH ２ ) ｊ -, wherein i is 1 to 10; j is 1 to 10; and R ３ The compound of any one of claims 1 to 9, or a pharma- ceutically acceptable salt or solvate thereof, wherein is hydrogen or alkyl.

11. X ４ but, 【Chemistry 3】 9. The compound according to any one of claims 1 to 8, which is: or a pharma- ceutically acceptable salt or solvate thereof.

12. Q is -C(O)-(CH ２ ) １－１０ - and L ４ is -C(O)NH-(CH ２ ) １－１０ 12. The compound of any one of claims 1 to 11, or a pharma- ceutically acceptable salt or solvate thereof, which is -phosphate.

13. Q is -C(O)-(CH ２ ) ３ - and L ４ is -C(O)NH-(CH ２ ) ６ 12. The compound of any one of claims 1 to 11, or a pharma- ceutically acceptable salt or solvate thereof, which is -phosphate.

14. L ４ The compound according to any one of claims 1 to 11, or a pharma- ceutically acceptable salt or solvate thereof, wherein: is -C(O)O-.

15. L ４ is -C(O)NH-(CH ２ ) １－１０ 12. The compound of any one of claims 1 to 11, or a pharma- ceutically acceptable salt or solvate thereof, which is -phosphate.

16. R １ is selected from the group consisting of pentafluorophenyl, tetrafluorophenyl, succinimide, maleimide, azide, pyridyldithiol, dimethyl methylphosphonate, chiral-dimethyl methylphosphonate, helper lipids, and nucleic acids, or a pharma- ceutically acceptable salt or solvate thereof.

17. R １ 16. The compound of any one of claims 1 to 15, or a pharma- ceutically acceptable salt or solvate thereof, wherein is ASO (antisense oligonucleotide), siRNA (small interfering RNA), miRNA (microRNA), microRNA mimic, AMO (anti-miRNA oligonucleotide), long non-coding RNA, PNA (peptide nucleic acid), helper lipid, or PMO (phosphorodiamidate morpholino oligomer), wherein said nucleic acid is optionally modified.

18. R １ The compound according to any one of claims 1 to 17, or a pharma- ceutically acceptable salt or solvate thereof, wherein is an ASO (antisense oligonucleotide).

19. formula: 【Chemistry 4】 having In the formula, R １ is an ASO (antisense oligonucleotide), an siRNA (small interfering RNA), an miRNA (microRNA), a microRNA mimic, an AMO (anti-miRNA oligonucleotide), a long non-coding RNA, a PNA (peptide nucleic acid), a helper lipid, or a PMO (phosphorodiamidate morpholino oligomer), or a pharma- ceutically acceptable salt or solvate thereof. 【Request 20】 【Chemical 5-1】 【Chemistry 5-2】 【Chemistry 5-3】 【Chemistry 5-4】 【Chemistry 5-5】 【Chemistry 5-6】 wherein: 【Chemistry 6】 is an oligonucleotide, or a pharma- ceutically acceptable salt or solvate thereof.

21. Compound of formula IB 【Chemistry 7】 or a pharma- ceutically acceptable salt or solvate thereof, wherein: X １ , X ２ and X ３ are each independently １ -C １０ Alkyl, -(CH ２ ) ｍ -O-(CH ２ ) ｎ - and - (CH ２ ) ｍ -NR Ｎ - (CH ２ ) ｎ -, wherein n is 1 to 36, m is 1 to 30, and R Ｎ is H, methyl, or CH ２ F, C.H.F. ２ , or C.F. ３ and Y １ , Y ２ and Y ３ each independently represents -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, or P(Z)(OH)O ２ wherein Z is O or S; L １ , L ２ and L ３ are each independently １ -C １０ Alkyl, -(CH ２ ) ｅ -O-(CH ２ ) ｆ -, -(CH ２ ) ｅ -S-(CH ２ ) ｆ -, -(CH ２ ) ｅ -S (O) ２ - (CH ２ ) ｆ -, -(CH ２ ) ｅ -NR Ｎ - (CH ２ ) ｆ - and - (CH ２ -CH ２ -O) ｋ (CH ２ ) ２ -, wherein e is 1 to 10; f is 1 to 16; k is 1 to 20; and R Ｎ is H, methyl, or CH ２ F, C.H.F. ２ , or C.F. ３ and G １ , G ２ and G ３ are each independently selected from the group consisting of monosaccharides and monosaccharide derivatives; X ４ is -NHC(O)R ２ and R ２ is carbocycle, heterocyclyl or heteroaryl, and R ２ is optionally substituted; Q is alkylamino, -C(O)-(CH ２ ) ｉ -, -(CH ２ ) ｉ -O-(CH ２ ) ｊ -, -(CH ２ ) ｉ -NR ３ - (CH ２ ) ｊ -, -(CH ２ ) ｉ -S-S-(CH ２ ) ｊ -, -(CH ２ ) ｉ -S-(CH ２ ) ｊ -, -(CH ２ ) ｉ -S (O) ２ - (CH ２ ) ｊ -, -(CH ２ ) ｉ -NHC(O)-(CH ２ ) ｊ -, -(CH ２ ) ｉ -C(O)NH-(CH ２ ) ｊ -, -(CH ２ ) ｉ -SC(O)-(CH ２ ) ｊ -, -(CH ２ ) ｉ -C(O)S-(CH ２ ) ｊ -,or 【Chemistry 8】 and In the formula, H １ is carbocycle, heterocyclyl or heteroaryl; H １ is optionally substituted; i is 1 to 30 and j is 1 to 36; R ３ is hydrogen or alkyl; W １ and W ２ Each independently represents -CH ２ - and O; v is 1 to 6; Y is hydrogen or methyl; and T is C １ -C １０ Alkyl or C ２ -C １０ alkenyl; L ４ is -C(O)O-, -C(O)NH-, phosphate, C １ -C １０ Alkyl-phosphate, C ３ -C １０ Alkenyl-phosphate, phosphorothioate, C １ -C １０ Alkyl-phosphorothioate, C ３ -C １０ Alkenyl-phosphorothioate, boranophosphate, C １ -C １０ Alkyl-boranophosphate, C ３ -C １０ Alkenyl-boranophosphate, —C(O)NH—C １ -C １０ Alkyl-phosphate, -C(O)NH-C ３ -C １０ Alkenyl-phosphate, —C(O)O—C １ -C １０ Alkyl-phosphate, -C(O)O-C ３ -C １０ Alkenyl-phosphate, -C(O)NH-C １ -C １０ Alkyl-phosphorothioates, -C(O)NH-C ３ -C １０ Alkenyl-phosphorothioate, -C(O)O-C １ -C １０ Alkyl-phosphorothioate, -C(O)O-C ３ -C １０ Alkenyl-phosphorothioate, -C(O)-NH-C １ -C １０ Alkyl-boranophosphate, —C(O)—NH—C ３ -C １０ Alkenyl-boranophosphate, —C(O)O—C １ -C １０ Alkyl-boranophosphate or -C(O)O-C ３ -C １０ alkenyl-boranophosphate; and R １ is a biologically active molecule, or a pharma- ceutically acceptable salt or solvate thereof.

22. Compound of Formula IC 【Chemistry 9】 or a pharma- ceutically acceptable salt or solvate thereof, wherein: X １ , X ２ and X ３ are each independently １ -C １０ Alkyl, -(CH ２ ) ｍ -O-(CH ２ ) ｎ - and - (CH ２ ) ｍ -NR Ｎ - (CH ２ ) ｎ -, wherein n is 1 to 36, m is 1 to 30, and R Ｎ is H, methyl, or CH ２ F, C.H.F. ２ , or C.F. ３ and Y １ , Y ２ and Y ３ each independently represents -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, or P(Z)(OH)O ２ wherein Z is O or S; L １ , L ２ and L ３ are each independently １ -C １０ Alkyl, -(CH ２ ) ｅ -O-(CH ２ ) ｆ -, -(CH ２ ) ｅ -S-(CH ２ ) ｆ -, -(CH ２ ) ｅ -S (O) ２ - (CH ２ ) ｆ -, -(CH ２ ) ｅ -NR Ｎ - (CH ２ ) ｆ - and - (CH ２ -CH ２ -O) ｋ (CH ２ ) ２ -, wherein e is 1 to 10, f is 1 to 16, k is 1 to 20, and R Ｎ is H, methyl, or CH ２ F, C.H.F. ２ , or C.F. ３ and G １ , G ２ and G ３ are each independently selected from the group consisting of monosaccharides and monosaccharide derivatives; X ４ is -NHC(O)R ２ and R ２ is carbocycle, heterocyclyl or heteroaryl, and wherein R ２ is optionally substituted; Q is, 【Chemistry 10-1】 【Chemistry 10-2】 In the formula, H １ is carbocycle, heterocyclyl or heteroaryl; H １ is optionally substituted; W １ and W ２ Each independently represents -CH ２ - and O; v is 1 to 6; wherein Y is hydrogen or methyl; and T is C １ -C １０ Alkyl or C １ -C １０ alkenyl; L ４ is -C(O)O-, -C(O)NH-, phosphate, C １ -C １０ Alkyl-phosphate, C ３ -C １０ Alkenyl-phosphate, phosphorothioate, C １ -C １０ Alkyl-phosphorothioate, C ３ -C １０ Alkenyl-phosphorothioate, boranophosphate, C １ -C １０ Alkyl-boranophosphate, C ３ -C １０ Alkenyl-boranophosphate, —C(O)NH—C １ -C １０ Alkyl-phosphate, -C(O)NH-C ３ -C １０ Alkenyl-phosphate, —C(O)O—C １ -C １０ Alkyl-phosphate, -C(O)O-C ３ -C １０ Alkenyl-phosphate, -C(O)NH-C １ -C １０ Alkyl-phosphorothioates, -C(O)NH-C ３ -C １０ Alkenyl-phosphorothioate, -C(O)O-C １ -C １０ Alkyl-phosphorothioate, -C(O)O-C ３ -C １０ Alkenyl-phosphorothioate, -C(O)-NH-C １ -C １０ Alkyl-boranophosphate, —C(O)—NH—C ３ -C １０ Alkenyl-boranophosphate, —C(O)O—C １ -C １０ Alkyl-boranophosphate or -C(O)O-C ３ -C １０ alkenyl-boranophosphate; and R １ is a biologically active molecule, or a pharma- ceutically acceptable salt or solvate thereof.

23. A compound according to any one of claims 1 to 22 or a pharma- ceutically acceptable salt or solvate thereof, and a lipid of formula II 【Chemistry 11】 or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony, R ５ and R ６ each independently represents a straight or branched C １ -C ３１ Alkyl, C ２ -C ３１ Alkenyl or C ２ -C ３１ Selected from the group consisting of alkynyl and cholesteryl; L ５ and L ６ each independently represents a linear C １ -C ２０ Alkyl and C ２ -C ２０ alkenyl; X ５ is —C(O)O— or —OC(O)—; X ６ is —C(O)O— or —OC(O)—; X ７ is S or O; L ７ is absent or lower alkyl; R ４ is a straight or branched C １ -C ６ is alkyl; and R ７ and R ８ are each independently hydrogen and a straight-chain or branched C １ -C ６ The pharmaceutical composition wherein said compound is selected from the group consisting of alkyl.

24. A compound according to any one of claims 1 to 22 or a pharma- ceutically acceptable salt or solvate thereof, and a lipid of formula III 【Chemistry 12】 or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony, R ９ and R １０ each independently represents a straight or branched C １－２０ Alkyl, linear or branched C ２ -C ２０ Alkenyl and C ２ -C ２０ alkynyl; L ８ and L ９ are each independently absent, linear C １ -C １８ Alkyl, or linear C ２ -C １８ alkenyl; L １１ is absent, a bond or a straight or branched C １ -C ６ is alkyl; L １０ is absent or methyl; X ８ is S or O; R １１ is a straight or branched C １ -C ６ is alkyl; and R １２ and R １３ are each independently hydrogen, and a straight or branched C １ -C ６ The pharmaceutical composition wherein said compound is selected from the group consisting of alkyl.

25. Lipid of Formula IV 【Chemistry 13】 or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony, X １ , X ２ and X ３ are each independently １ -C １０ Alkyl, -(CH ２ ) ｍ -O-(CH ２ ) ｎ - and - (CH ２ ) ｍ -NR Ｎ - (CH ２ ) ｎ -, wherein n is 1 to 36, m is 1 to 30, and R Ｎ is H, methyl, or CH ２ F, C.H.F. ２ , or C.F. ３ and Y １ , Y ２ and Y ３ each independently represents -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, or P(Z)(OH)O ２ wherein Z is O or S; L １ , L ２ and L ３ are each independently １ -C １０ Alkyl, -(CH ２ ) ｅ -O-(CH ２ ) ｆ -, -(CH ２ ) ｅ -S-(CH ２ ) ｆ -, -(CH ２ ) ｅ -S (O) ２ - (CH ２ ) ｆ - and - (CH ２ ) ｅ -NR Ｎ - (CH ２ ) ｆ -, -(CH ２ -CH ２ -O)e(CH ２ ) ２ -, wherein e is 1 to 10, f is 1 to 16, k is 1 to 20, and R Ｎ is H, methyl, or CH ２ F, C.H.F. ２ , or C.F. ３ and G １ , G ２ and G ３ are each independently selected from the group consisting of monosaccharides and monosaccharide derivatives; X ４ is -NHC(O)R ２ and R ２ is carbocycle, heterocyclyl or heteroaryl, and R ２ is optionally substituted; Q is alkylamino, -O(O)C-, -C(O)O-, -NHC(O)-, -C(O)NH-, -(CH ２ ) ｊ -NHC(O)-, -C(O)NH-(CH ２ ) ｊ -, -C(O)-(CH ２ ) ｉ -, -(CH ２ ) ｉ -O-(CH ２ ) ｊ -, -(CH ２ ) ｉ -NR ３ - (CH ２ ) ｊ -, -(CH ２ ) ｉ -S-S-(CH ２ ) ｊ -, -(CH ２ ) ｉ -S-(CH ２ ) ｊ -, -(CH ２ ) ｉ -S (O) ２ - (CH ２ ) ｊ -, -(CH ２ ) ｉ -NHC(O)-(CH ２ ) ｊ -, -(CH ２ ) ｉ -C(O)NH-(CH ２ ) ｊ -, -(CH ２ ) ｉ -SC(O)-(CH ２ ) ｊ - or - (CH ２ ) ｉ -C(O)S-(CH ２ ) ｊ -, where i is 1 to 30, j is 1 to 36, and where R ３ is hydrogen or alkyl; L ４ is -PEG-C(O)O-, PEG-C(O)NH-, -PEG-NHC(O)-, -PEG-phosphate, -PEG-C １ -C １０ Alkyl-phosphate, -PEG-C ３ -C １０ Alkenyl-phosphate, -PEG-phosphorothioate, -PEG-C １ -C １０ Alkyl-phosphorothioate, -PEG-C ３ -C １０ Alkenyl-phosphorothioate, -PEG-boranophosphate, -PEG-C １ -C １０ Alkyl-boranophosphate, -PEG-C ３ -C １０ Alkenyl-boranophosphate, -PEG-C(O)NH-C １ -C １０ Alkyl-phosphate, -PEG-C(O)NH-C ３ -C １０ Alkenyl-phosphate, -PEG-C(O)O-C １ -C １０ Alkyl-phosphate, -PEG-C(O)O-C ３ -C １０ Alkenyl-phosphate, -PEG-C(O)NH-C １ -C １０ Alkyl-phosphorothioate, -PEG-C(O)NH-C ３ -C １０ Alkenyl-phosphorothioate, -PEG-C(O)O-C １ -C １０ Alkyl-phosphorothioate, -PEG-C(O)O-C ３ -C １０ Alkenyl-phosphorothioate, -PEG-C(O)-NH-C １ -C １０ Alkyl-boranophosphate, -PEG-C(O)-NH-C ３ -C １０ Alkenyl-boranophosphate, -PEG-C(O)O-C １ -C １０ Alkyl-boranophosphate or -PEG-C(O)O-C ３ -C １０ alkenyl-boranophosphate; and R １ is a helper lipid.

26. A pharmaceutical composition according to any one of claims 23 to 25 for use in the treatment of a disease.

27. 23. A pharmaceutical composition comprising a compound according to any one of claims 1 to 22, or a pharma- ceutically acceptable salt or solvate thereof, for use in the treatment of a disease.

28. 28. The pharmaceutical composition according to claim 26 or 27, for parenteral administration or intravenous injection.

29. The pharmaceutical composition according to any one of claims 26 to 28, which is for subcutaneous, intradermal or intramuscular injection.

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