LIPID NANOPARTICLES

1 . A lipid nanoparticle comprising:

(a) a nucleic acid:

(b) a cationic lipid in an amount from about 30 to about 70 mol % of the total lipid present in the particle;

(c) a non-cationic lipid in an amount from about 30 to about 70 mol % of the total lipid present in the particle, wherein the non-cationic lipid comprises a mixture of a phospholipid and a cholesterol or derivative thereof; and

(d) a polyethylene glycol (PEG)-lipid conjugate that inhibits aggregation of lipid nanoparticles in an amount from about 2 to about 5 mol % of the total lipid in the particle,

wherein the PEG-lipid conjugate comprises a PEG moiety linked to a lipid anchor moiety,

wherein the PEG moiety of the PEG-lipid conjugate has an average molecular weight of from about 500 to about 1,000 daltons,

provided that when the lipid anchor moiety is a dialkyl moiety, at least one of the two alkyl chains is less than C14.

2 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety is a single C 10 -C 24 alkyl chain.

3 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety is a single C 10 , C 12 , C 14 , C 16 or C 18 chain.

4 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety is a single C 18 chain.

5 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety is a single C 16 -C 24 alkyl chain.

6 . The lipid nanoparticle of claim 5 , wherein the lipid anchor moiety is a single C 18 -C 22 alkyl chain.

7 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety comprises a sterol or sterol derivative.

8 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety comprises cholesterol or a cholesterol derivative.

9 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety comprises a polycyclic structure.

10 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety is a dialkyl moiety.

11 . The lipid nanoparticle of claim 10 , wherein the lipid anchor moiety is a symmetric dialkyl moiety.

12 . The lipid nanoparticle of claim 10 , wherein the lipid anchor moiety is an asymmetric dialkyl moiety.

13 . The lipid nanoparticle of claim 12 , wherein the lipid anchor moiety is an asymmetric dialkyl moiety having C 10 and C 14 alkyl chains.

14 . The lipid nanoparticle of claim 11 , wherein the lipid anchor moiety is a dialkyl moiety having two CR alkyl chains.

15 . The lipid nanoparticle of claim 11 , wherein the lipid anchor moiety is a dialkyl moiety having two C 10 alkyl chains.

16 . The lipid nanoparticle of claim 11 , wherein the lipid anchor moiety is a dialkyl moiety having two C 12 alkyl chains.

17 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety is a trialkyl moiety.

18 . The lipid nanoparticle of claim 17 , wherein the lipid anchor moiety is a trialkyl moiety having three alkyl chains of C 10 or less.

19 . The lipid nanoparticle of claim 17 or 18 , wherein the lipid anchor moiety is a symmetric trialkyl moiety.

20 . The lipid nanoparticle of claim 17 or 18 , wherein the lipid anchor moiety is an asymmetric trialkyl moiety.

21 . The lipid nanoparticle of claim 1 , wherein the lipid anchor moiety is a tetraalkyl moiety.

22 . The lipid nanoparticle of claim 21 , wherein the lipid anchor moiety is a tetraalkyl moiety having three alkyl chains of C 8 or less.

23 . The lipid nanoparticle of claim 21 or 22 , wherein the lipid anchor moiety is a symmetric tetraalkyl moiety.

24 . The lipid nanoparticle of claim 21 or 22 , wherein the lipid anchor moiety is an asymmetric tetraalkyl moiety.

25 . The lipid nanoparticle of any one of claims 1 - 24 , wherein the PEG moiety of the PEG-lipid conjugate has an average molecular weight of about 750 daltons.

26 . The lipid nanoparticle of any one of claims 1 - 25 , wherein the polyethylene glycol (PEG)-lipid conjugate that inhibits aggregation of lipid nanoparticles is present in an amount of about 2 mol % of the total lipid in the particle.

27 . The lipid nanoparticle of any one of claims 1 - 25 , wherein the polyethylene glycol (PEG)-lipid conjugate that inhibits aggregation of lipid nanoparticles is present in an amount of about 3 mol % of the total lipid in the particle.

28 . The lipid nanoparticle of any one of claims 1 - 25 , wherein the polyethylene glycol (PEG)-lipid conjugate that inhibits aggregation of lipid nanoparticles is present in an amount of about 4 mol % of the total lipid in the particle.

29 . The lipid nanoparticle of any one of claims 1 - 25 , wherein the polyethylene glycol (PEG)-lipid conjugate that inhibits aggregation of lipid nanoparticles is present in an amount of about 5 mol % of the total lipid in the particle.

30 . A lipid nanoparticle comprising:

(a) a nucleic acid;

(b) a cationic lipid in an amount from about 30 to about 70 mol % of the total lipid present in the particle;

(c) a non-cationic lipid in an amount from about 30 to about 70 mol % of the total lipid present in the particle, wherein the non-cationic lipid comprises a mixture of a phospholipid and a cholesterol or derivative thereof; and

(d) a polyethylene glycol (PEG)-lipid conjugate that inhibits aggregation of lipid nanoparticles in an amount from about 0.2 to about 0.5 mol % of the total lipid in the particle,

wherein the PEG-lipid conjugate comprises a PEG moiety linked to a lipid anchor moiety,

wherein the PEG moiety of the PEG-lipid conjugate has an average molecular weight of from about 5,000 to about 20,000 daltons.

31 . The lipid nanoparticle of claim 30 , wherein the lipid anchor moiety is a single alkyl chain.

32 . The lipid nanoparticle of claim 30 , wherein the lipid anchor moiety comprises a sterol or sterol derivative.

33 . The lipid nanoparticle of claim 30 , wherein the lipid anchor moiety comprises cholesterol or a cholesterol derivative.

34 . The lipid nanoparticle of claim 30 , wherein the lipid anchor moiety comprises a polycyclic structure.

35 . The lipid nanoparticle of claim 30 , wherein the lipid anchor moiety is a dialkyl moiety.

36 . The lipid nanoparticle of claim 35 , wherein the lipid anchor moiety is a symmetric dialkyl moiety.

37 . The lipid nanoparticle of claim 35 , wherein the lipid anchor moiety is an asymmetric dialkyl moiety.

38 . The lipid nanoparticle of claim 35 , wherein the lipid anchor moiety is a dialkyl moiety having alkyl chains longer than C14.

39 . The lipid nanoparticle of claim 35 , wherein the lipid anchor moiety is a dialkyl moiety having C 14 -C 22 alkyl chains.

40 . The lipid nanoparticle of claim 36 , wherein the lipid anchor moiety is a dialkyl moiety having two C 14 alkyl chains.

41 . The lipid nanoparticle of claim 36 , wherein the lipid anchor moiety is a dialkyl moiety having two C 16 alkyl chains.

42 . The lipid nanoparticle of claim 36 , wherein the lipid anchor moiety is a dialkyl moiety having two C 18 alkyl chains.

43 . The lipid nanoparticle of claim 30 , wherein the lipid anchor moiety is a trialkyl moiety.

44 . The lipid nanoparticle of claim 30 , wherein the lipid anchor moiety is a trialkyl moiety having three alkyl chains of C 8 or greater.

45 . The lipid nanoparticle of claim 43 or 44 , wherein the lipid anchor moiety is a symmetric trialkyl moiety.

46 . The lipid nanoparticle of claim 43 or 44 , wherein the lipid anchor moiety is an asymmetric trialkyl moiety.

47 . The lipid nanoparticle of claim 30 , wherein the lipid anchor moiety is a tetraalkyl moiety.

48 . The lipid nanoparticle of claim 47 , wherein the lipid anchor moiety is a tetraalkyl moiety having three alkyl chains of C 6 or greater.

49 . The lipid nanoparticle of claim 47 or 48 , wherein the lipid anchor moiety is a symmetric tetraalkyl moiety.

50 . The lipid nanoparticle of claim 47 or 48 , wherein the lipid anchor moiety is an asymmetric tetraalkyl moiety.

51 . The lipid nanoparticle of any one of claims 30 - 50 , wherein the PEG moiety of the PEG-lipid conjugate has an average molecular weight of from about 5,000 to about 10,000 daltons.

52 . The lipid nanoparticle of any one of claims 30 - 50 , wherein the PEG moiety of the PEG-lipid conjugate has an average molecular weight of from about 8,000 to about 10,000 daltons.

53 . The lipid nanoparticle of any one of claims 30 - 50 , wherein the PEG moiety of the PEG-lipid conjugate has an average molecular weight of about 5,000 daltons.

54 . The lipid nanoparticle of any one of claims 30 - 50 , wherein the PEG moiety of the PEG-lipid conjugate has an average molecular weight of about 10,000 daltons.

55 . The lipid nanoparticle of any one of claims 1 - 54 , wherein the nucleic acid is at least 80 bases in length.

56 . The lipid nanoparticle of any one of claims 1 - 54 , wherein the nucleic acid is at least 100 bases in length.

57 . The lipid nanoparticle of any one of claims 1 - 54 , wherein the nucleic acid is at least 500 bases in length.

58 . The lipid nanoparticle of any one of claim 1 - 54 , wherein the nucleic acid is DNA, plasmid DNA, minicircle DNA, ceDNA (closed ended DNA), mRNA, self-replicating RNA, CRISPR RNA, a gene editing construct, an RNA editing construct, or a base editing construct.

59 . The lipid nanoparticle of any one of claim 1 - 54 , wherein the nucleic acid is mRNA.

60 . The lipid nanoparticle of any one of claims 1 - 54 , wherein the nucleic acid is siRNA.

61 . The lipid nanoparticle of any one of claims 1 - 54 , wherein the nucleic acid is not siRNA.

62 . The lipid nanoparticle of any one of claims 1 - 61 , wherein the cationic lipid is a compound of formula CL 1 or a salt thereof.

63 . The lipid nanoparticle of any one of claims 1 - 61 , wherein the cationic lipid is a compound of formula CL 2 or a salt thereof:

64 . The lipid nanoparticle of any one of claims 1 - 61 , wherein the cationic lipid is a compound of formula CL 3 or a salt thereof:

65 . The nucleic acid-lipid particle of any one of claims 1 - 64 , wherein the phospholipid is DSPC.

66 . The nucleic acid-lipid particle of any one of claims 1 - 65 , wherein the lipid anchor moiety comprises at least one saturated alkyl chain.

67 . The nucleic acid-lipid particle of any one of claims 1 - 66 , wherein the lipid anchor moiety comprises at least one unsaturated alkyl chain.

68 . The nucleic acid-lipid particle of any one of claims 1 - 67 , wherein the lipid anchor moiety comprises at least one alkyl chain having at least one double bond.

69 . A lipid nanoparticle comprising:

(a) a nucleic acid, wherein the nucleic acid is mRNA:

(b) a cationic lipid:

(c) a non-cationic lipid, wherein the non-cationic lipid comprises a mixture of a phospholipid and a cholesterol or derivative thereof; and

(d) a polyethylene glycol (PEG)-lipid conjugate that inhibits aggregation of lipid nanoparticles,

wherein the PEG-lipid conjugate comprises a PEG moiety linked to a lipid anchor moiety, wherein the lipid anchor moiety is a single chain C 18 alkyl moiety.

70 . A lipid nanoparticle comprising:

(a) a nucleic acid, wherein the nucleic acid is mRNA;

(b) a cationic lipid in an amount from about 30 to about 70 mol % of the total lipid present in the particle;

(c) a non-cationic lipid in an amount from about 30 to about 70 mol % of the total lipid present in the particle, wherein the non-cationic lipid comprises a mixture of a phospholipid and a cholesterol or derivative thereof; and

(d) a polyethylene glycol (PEG)-lipid conjugate that inhibits aggregation of lipid nanoparticles in an amount from about 1 to about 2.5 mol % of the total lipid in the particle (e.g., about 1.6 mol %),

wherein the PEG-lipid conjugate comprises a PEG moiety linked to a lipid anchor moiety, wherein the lipid anchor moiety is a single chain C 14 -C 22 (e.g., C 14 , C 16 , C 18 , C 20 or C 22 ) alkyl moiety,

wherein the PEG moiety of the PEG-lipid conjugate has an average molecular weight of from about 500 to about 3,000 daltons (e.g., about 2000 daltons).

71 . A pharmaceutical composition comprising a lipid nanoparticle as described in any one of claims 1 - 70 , and a pharmaceutically acceptable carrier.

72 . The pharmaceutical composition of claim 71 , which is formulated for intravenous administration.

73 . A method for delivering a nucleic acid to a cell comprising contacting the cell with a lipid nanoparticle as described in any one of claims 1 - 70 .

74 . A method for treating a disease characterized by a genetic defect that results in a deficiency of a functional protein, the method comprising: administering to a subject having the disease, a lipid nanoparticle as described in claim 59 , wherein the mRNA encodes the functional protein or a protein having the same biological activity as the functional protein.

75 . A method for treating a disease characterized by overexpression of a polypeptide, comprising administering to a subject having the disease a lipid nanoparticle as described in claim 57 , wherein the siRNA targets expression of the overexpressed polypeptide.

76 . A lipid nanoparticle as described in any one of claims 1 - 70 , for the therapeutic or prophylactic treatment of a disease characterized by a genetic defect that results in a deficiency of a functional protein.

77 . A lipid nanoparticle as described in any one of claims 1 - 70 , for the therapeutic or prophylactic treatment of a disease characterized by overexpression of a polypeptide.

78 . A method for reducing the immune response of administration of a lipid nanoparticle (LNP) to a human, comprising selecting a polyethylene glycol (PEG)-lipid conjugate for use in LNP, wherein the PEG-lipid conjugate comprises a PEG moiety linked to a lipid anchor moiety, wherein the lipid anchor moiety is a single alkyl chain, wherein the LNP comprises an mRNA payload.

79 . The method of claim 78 , wherein the lipid anchor moiety is a single C 10 -C 24 alkyl chain.

80 . The method of claim 79 , wherein the lipid anchor moiety is a single C 10 , C 12 , C 14 , C 16 C 18 C 20 , or C 22 , chain.

81 . The method of claim 80 , wherein the lipid anchor moiety is a single C 18 chain.

82 . The method of any one of claims 78 - 81 , further comprising treating a human in need thereof with an initial administration of the LNP and at least one subsequent administration of the LNP.