Compositions and methods for organ specific delivery of nucleic acids

1. A composition comprising a SORT lipid composition assembled with a therapeutic agent, which SORT lipid composition comprises:

(1) An ionizable cationic lipid; and

(2) A permanently cationic selective organ targeting (SORT) lipid,

wherein said SORT lipid composition comprises said permanently cationic SORT lipid at a molar percentage of about 5% or more, wherein the molar percentage is determined based on total mols of lipids present in the SORT lipid composition;

wherein said SORT lipid composition is characterized by an apparent pKa of about 8 or higher as determined by a 2-(p-toluidino)-6-naphthalenesulfonic acid titration assay; and

wherein said composition delivers said therapeutic agent to a lung or a lung cell following intravenous administration or inhalation;

wherein (1) the ionizable cationic lipid is a compound having the structure of Formula (I)

Core-Repeating Unit-Terminating  Group (I)

or a pharmaceutically acceptable salt thereof,

wherein the compound of Formula (I) or the pharmaceutically acceptable salt is a dendron or dendrimer, wherein

the Core is made from the structure of Formula (II) which has been linked to one or more Repeating Unit(s) by removing one or more hydrogen atoms from the structure of Formula (II) and replacing the one or more hydrogen atoms with the Repeating Unit(s) to form the Core, wherein Formula (II) is

wherein, in Formula (II):

X 1 is amino, optionally substituted alkylamino (C≤12) , optionally substituted dialkylamino (C≤12) , optionally substituted heterocycloalkyl (C≤12) , or optionally substituted heteroaryl (C≤12) ;

R 1 is amino, hydroxy, mercapto, optionally substituted alkylamino (C≤12) , or optionally substituted dialkylamino (C≤12) ; and

a is 1, 2, 3, 4, 5, or 6; or

alternatively, the Core is made from a compound with the structure of Formula (III) which has been linked to one or more Repeating Unit(s) by removing one or more hydrogen atoms from the structure of Formula (III) and replacing the one or more hydrogen atoms with the Repeating Unit(s) to form the Core, wherein Formula (III) is:

wherein, in Formula (III):

X 2 is N(R 5 ) y , wherein:

R 5 is hydrogen or optionally substituted alkyl (C≤18) ; and

y is 0, 1, or 2, provided that the sum of y and z is 3;

R 2 is amino, hydroxy, mercapto, optionally substituted alkylamino (C≤12) , or optionally substituted dialkylamino (C≤12) ;

b is 1, 2, 3, 4, 5, or 6; and

z is 1, 2, 3; provided that the sum of z and y is 3; or

alternatively, the Core is made from a compound with the structure of Formula (IV) which has been linked to one or more Repeating Unit(s) by removing one or more hydrogen atoms from the structure of Formula (IV) and replacing the one or more hydrogen atoms with the Repeating Unit(s) to form the Core, wherein Formula (IV) is:

wherein, in Formula (IV):

X 3 is selected from —NR 6 —, —O—, optionally substituted alkylaminodiyl (C≤8) , optionally substituted alkoxydiyl (C≤8) , optionally substituted arenediyl (C≤8) , optionally substituted heteroarenediyl (C≤8) , and optionally substituted heterocycloalkanediyl (C≤8) ,

wherein R 6 is hydrogen, alkyl (C≤8) , or substituted alkyl (C≤8) ;

R 3 and R 4 are each independently selected from amino, hydroxy, mercapto, optionally substituted alkylamino (C≤12) , and optionally substituted dialkylamino (C≤12) ; and

c and d are each independently 1, 2, 3, 4, 5, or 6; or

alternatively, the Core is made from an optionally substituted alkylamine (C≤18) , an optionally substituted dialkylamine (C≤36) , or an optionally substituted heterocycloalkane (C≤12) which has been linked to one or more Repeating Unit(s) by removing one or more hydrogen atom and replacing the one or more hydrogen atoms with the Repeating Unit(s) to form the Core;

the Repeating Unit(s) comprises a degradable diacyl and optionally a linker; wherein:

the degradable diacyl group has the formula:

wherein, in Formula (VII): A 1 and A 2 are each independently —O— or —NR a — wherein R a is hydrogen or optionally substituted alkyl (C≤6) ;

Y 3 is selected from optionally substituted alkanediyl (C≤12) , optionally substituted alkenediyl (C≤12) , optionally substituted arenediyl (C≤12) , and a group of the formula:

X 3A and X 4 are each independently selected from optionally substituted alkanediyl (C≤12) , optionally substituted alkenediyl (C≤12) , and optionally substituted arenediyl (C≤12) ; and

Y 5 is selected from a covalent bond, optionally substituted alkanediyl (C≤12) , optionally substituted alkenediyl (C≤12) , and optionally substituted arenediyl (C≤12) ; and

R 9 is optionally substituted alkyl (C≤8) ; and

the linker group has the formula:

Y 1 is selected from optionally substituted alkanediyl (C≤12) , optionally substituted alkenediyl (C≤12) , and optionally substituted arenediyl (C≤12) ; and

wherein when the repeating unit comprises the linker group, then the linker group is attached to the degradable diacyl group on both the nitrogen and the sulfur atoms of the linker group, wherein the first group in the repeating unit is the degradable diacyl group, wherein for each linker group, the next group comprises two degradable diacyl groups attached to the nitrogen atom of the linker group; and

wherein n is the number of linker groups present in the repeating unit; wherein n is 0, 1, 2, 3, 4, 5, or 6; and

the Terminating Group has the formula:

wherein, in Formula (VIII):

Y 4 is alkanediyl (C≤8) optionally substituted with one or more substituents independently selected from —OH, —F, —Cl, —Br, —I, —SH, —OCH 3 , —OCH 2 CH 3 , —SCH 3 , and —OC(O)CH 3 ;

R 10 is selected from hydrogen, carboxy, hydroxy, aryl (C≤12) , alkylamino (C≤12) , dialkylamino (C≤12) , N-heterocycloalkyl (C≤12) , —C(O)N(R 11 )-alkanediyl (C≤6) -heterocycloalkyl (C≤12) , —C(O)-alkylamino (C≤12) , —C(O)-dialkylamino (C≤12) , and —C(O)—N-heterocycloalkyl (C≤12) , wherein:

R 11 is hydrogen or optionally substituted alkyl (C≤6) ; and

wherein the final degradable diacyl in the chain of repeating unit(s) is attached to the Terminating Group; and

wherein (2) the permanently cationic SORT lipid comprises (i) a cationic headgroup and (ii) a plurality of hydrophobic tails, wherein:

(i) the cationic headgroup comprises a quaternary ammonium moiety; and

(ii) the plurality of hydrophobic tails comprises from 2 to 6 hydrophobic tails, wherein a hydrophobic tail in the plurality of hydrophobic tails is independently C 6 -C 24 alkyl or C 6 -C 24 alkenyl.

2. The composition of claim 1 , wherein said permanently cationic SORT lipid is present in said SORT lipid composition at a molar percentage from about 5% to about 65%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.

3. The composition of claim 1 , wherein said permanently cationic SORT lipid has a structure of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt, stereoisomer, tautomer thereof:

wherein, in Formula (I):

R 1 and R 2 are each independently alkyl (C8-C24) , alkenyl (C8-C24) , or a substituted version of either group;

R 3 , R 3 ′, and R 3 ″ are each independently alkyl (C≤6) or substituted alkyl (C≤6) ; and

X − is a monovalent anion; or

wherein, in Formula (II):

R 4 and R 4 ′ are each independently alkyl (C6-C24) , alkenyl (C6-C24) , or a substituted version of either group;

R 4 ″ is alkyl (C≤24) , alkenyl (C≤24) , or a substituted version of either group;

R 4 ′″ is alkyl (C1-C8) , alkenyl (C2-C8) , or a substituted version of either group; and

X 2 − is a monovalent anion; or

wherein, in Formula (III):

R 1 and R 2 are each independently alkyl (C8-C24) , alkenyl (C8-C24) , or a substituted version of either group;

R 3 , R 3 ′, and R 3 ″ are each independently alkyl (C≤6) or substituted alkyl (C≤6) ;

R 4 is alkyl (C≤6) or substituted alkyl (C≤6) ; and

X − is a monovalent anion.

4. The composition of claim 1 , wherein said SORT lipid composition comprises said ionizable cationic lipid at a molar percentage from about 5% to about 30%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.

5. The composition of claim 1 , wherein said SORT lipid composition further comprises a zwitterionic lipid.

6. The composition of claim 5 , wherein said zwitterionic lipid is a zwitterionic phospholipid.

7. The composition of claim 5 , wherein said SORT lipid composition comprises said zwitterionic lipid at a molar percentage from about 8% to about 23%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.

8. The composition of claim 1 , wherein said SORT lipid composition further comprises a polymer-conjugated lipid.

9. The composition of claim 8 , wherein said SORT lipid composition comprises said polymer-conjugated lipid at a molar percentage from about 0.5% to about 10%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.

10. The composition of claim 8 , wherein said polymer-conjugated lipid is a poly(ethylene glycol) (PEG)-conjugated lipid.

11. The composition of claim 1 , wherein said SORT lipid composition further comprises a steroid or steroid derivative.

12. The composition of claim 11 , wherein said SORT lipid composition comprises said steroid or steroid derivative at a molar percentage from about 15% to about 46%, wherein the molar percentage is determined based on total lipids present in the SORT lipid composition.

13. The composition of claim 1 , wherein said therapeutic agent comprises a small interfering ribonucleic acid (siRNA), a micro-ribonucleic acid (miRNA), a primary micro-ribonucleic acid (pri-miRNA), a messenger ribonucleic acid (mRNA), a clustered regularly interspaced short palindromic repeats (CRISPR) related nucleic acid, a CRISPR-RNA (crRNA), a single guide ribonucleic acid (sgRNA), a trans-activating CRISPR ribonucleic acid (tracrRNA), a plasmid deoxyribonucleic acid (pDNA), a transfer ribonucleic acid (tRNA), an antisense oligonucleotide (ASO), a guide ribonucleic acid, a double stranded deoxyribonucleic acid (dsDNA), a single stranded deoxyribonucleic acid (ssDNA), a single stranded ribonucleic acid (ssRNA), a double stranded ribonucleic acid (dsRNA), a protein, a CRSIPR-associated (Cas) protein, or a combination thereof.

14. The composition of claim 1 , wherein said therapeutic agent comprises a messenger ribonucleic acid (mRNA).

15. The composition of claim 14 , wherein said mRNA encodes a Cas protein.

16. The composition of claim 14 , wherein said therapeutic agent further comprises a CRISPR-RNA (crRNA), a single guide ribonucleic acid (sgRNA), a trans-activating CRISPR ribonucleic acid (tracrRNA), or a combination thereof.

17. The composition of claim 14 , wherein said therapeutic agent further comprises a single guide polynucleotide.

18. The composition of claim 14 , wherein said therapeutic agent further comprises a double stranded deoxyribonucleic acid (dsDNA), a single stranded deoxyribonucleic acid (ssDNA), or a combination thereof.

19. The composition of claim 1 , wherein said therapeutic agent comprises a protein.

20. The composition of claim 19 , wherein said protein is a CRSIPR-associated (Cas) protein.

21. The composition of claim 19 , wherein said therapeutic agent further comprises a CRISPR-RNA (crRNA), a single guide ribonucleic acid (sgRNA), a trans-activating CRISPR ribonucleic acid (tracrRNA), or a combination thereof.

22. The composition of claim 19 , wherein said therapeutic agent further comprises a single guide polynucleotide.

23. The composition of claim 19 , wherein said therapeutic agent further comprises a double stranded deoxyribonucleic acid (dsDNA), a single stranded deoxyribonucleic acid (ssDNA), or a combination thereof.

24. The composition of claim 1 , wherein the permanently cationic SORT lipid comprises a trimethylammonium-propane (TAP) headgroup, or an ethylphosphocholine (EPC) headgroup.

25. The composition of claim 1 , wherein, in the permanently cationic SORT lipid, the plurality of hydrophobic tails comprises one or more saturated hydrophobic tails.

26. The composition of claim 1 , wherein, in the permanently cationic SORT lipid, the plurality of hydrophobic tails comprises one or more unsaturated hydrophobic tails.

27. The composition of claim 1 , wherein, in the permanently cationic SORT lipid, the plurality of hydrophobic tails is attached to the cationic headgroup through one or more linker moieties.

28. The composition of claim 27 , wherein, in the permanently cationic SORT lipid, the one or more linker moieties comprise one or more ester groups.

29. The composition of claim 1 , wherein said SORT lipid composition is characterized by an apparent pKa is of about 9 or higher as determined by a 2-(p-toluidino)-6-naphthalenesulfonic acid titration assay.

30. The composition of claim 1 , wherein said SORT lipid composition is characterized by an apparent pKa is from about 8 to about 13 as determined by a 2-(p-toluidino)-6-naphthalenesulfonic acid titration assay.

31. The composition of claim 1 , wherein, in the compound of Formula (I) or the pharmaceutically acceptable salt thereof, the Core is made from the structure of Formula (IV):

32. The composition of claim 1 , wherein, in the compound of Formula (I), or the pharmaceutically acceptable salt thereof, the Core is made from a structure selected from the group consisting of:

which is linked to the one or more Repeating Unit(s) by removing one or more hydrogen atoms and replacing the one or more hydrogen atoms with the Repeating Unit(s) to form the Core.

33. The composition of claim 1 , wherein, in the Terminating Group of Formula (VIII), Y 4 is alkanediyl (C≤18) ; and R 10 is hydrogen.