Granted Patent B2

US 12,344,709 · App. 17/051,157 · Granted Jul 1, 2025

Inventors: Kunwoo Lee (Berkeley, CA); Santanu Maity (Berkeley, CA)

Assignee: GenEdit Inc.

C08G69/10A61K9/5146A61K47/10A61K47/6455A61K47/6931C08G69/04C08G69/48

Abstract

Claims (106)

1. A polymer comprising a structure of Formula 1:

wherein:

each of m 1 and n 1 is an integer from 0 to 1000;

each of m 2 and n 2 is an integer from 0 to 1000, provided that the sum of m 2 +n 2 is greater than 5;

the symbol “/” indicates that the units separated thereby are linked randomly or in any order;

R 3a and R 3b are each independently a methylene or ethylene group;

each instance of R 13 is independently hydrogen or a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group;

A 1 and A 2 are each independently a group of formula

—(CH 2 ) p1 —[NR 2 —(CH 2 ) q1 -] r1 NR 2 2 ;

—(CH 2 ) p2 —N[—(CH 2 ) q2 —NR 2 2 ] 2 ;

—(CH 2 ) p3 —N{[—(CH 2 ) q3 —NR 2 2 ][—(CH 2 ) q4 —NR 2 -]r 2 R 2 }; or

—(CH 2 ) p4 —N{—(CH 2 ) q5 —N[—(CH 2 ) q6 —NR 2 2 ] 2 } 2 ,

B 1 and B 2 are each independently

—(CH 2 ) p1 —[NR 2 —(CH 2 ) q1 -] r1 NR 2 —(CH 2 ) s1 —R 4 —R 5 ;

—(CH 2 ) p2 —N[—(CH 2 ) q2 —NR 2 —(CH 2 ) s2 —R 4 —R 5 ] 2 ;

—(CH 2 ) p3 —N{[—(CH 2 ) q3 —NR 2 2 ][—(CH 2 ) q4 —NR 2 —] r2 (CH 2 ) s3 —R 4 —R 5 };

—(CH 2 ) p4 —N{—(CH 2 ) q5 —N[—(CH 2 ) q6 —NR 2 —(CH 2 ) s4 —R 4 —R 5 ] 2 } 2 ;

—(CH 2 ) p1 —[NR 2 —(CH 2 ) q1 -] r1 NR 2 —CH 2 —CHOH—R 5 ;

—(CH 2 ) p2 —N[—(CH 2 ) q2 —NR 2 —CH 2 —CHOH—R 5 ] 2 ;

—(CH 2 ) p3 —N{[—(CH 2 ) q3 —NR 2 2 ][—(CH 2 ) q4 —NR 2 —] r2 —CH 2 —CHOH—R 5 };

—(CH 2 ) p4 —N{—(CH 2 ) q5 —N[—(CH 2 ) q6 —NR 2 —CH 2 —CHOH—R 5 ] 2 } 2 ;

—(CH 2 ) p2 —N[—(CH 2 ) q2 —NR 2 —(CH 2 ) s2 —R 5 ] 2 ;

—(CH 2 ) p3 —N{[—(CH 2 ) q3 —NR 2 2 ][—(CH 2 ) q4 —NR 2 —] r2 (CH 2 ) s3 —R 5 };

—(CH 2 ) p4 —N{—(CH 2 ) q5 —N[—(CH 2 ) q6 —NR 2 —(CH 2 ) s4 —R 5 ] 2 } 2 ;

—(CH 2 ) p1 —[N{(CH 2 ) s1 —R 4 —R 5 }—(CH 2 ) q1 —] r1 NR 2 2 ; or

—(CH 2 ) p1 —[N{(CH 2 ) s1 —R 5 }—(CH 2 ) q1 -] r1 NR 2 2 ,

wherein p1 to p4, q1 to q6, r1 and r2, and s1 to s4 are each independently an integer of 1 to 5;

each instance of R 2 is independently hydrogen or a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group; each instance of R 4 is independently —C(O)O—, —C(O) NH—, or —S(O)(O)—; and each instance of R 5 is independently an alkyl group, cycloalkyl group, alkenyl group, cycloalkenyl group, aryl group, heteroalkyl group, heterocyclic group, or combination thereof optionally comprising from 1 to 8 secondary or tertiary amines or a substituent comprising a tissue-specific or cell-specific targeting moiety.

2. The polymer of claim 1 , wherein each of A 1 and A 2 is a group of formula —(CH 2 ) 2 —NH—(CH 2 ) 2 —NH 2 , and each of B 1 and B 2 is a group of formula —(CH 2 ) 2 —NH—(CH 2 ) 2 —NH—(CH 2 ) 2 —R 4 —R 5 .

3. The polymer of claim 1 having the structure of Formula 1A:

wherein

c is an integer from 0 to 50;

Y is optionally present and is a cleavable linker;

each of m 1 and n 1 is an integer from 0 to 1000;

each of m 2 and n 2 is an integer from 0 to 1000, provided that the sum of m 2 +n 2 is greater than 5;

the symbol “/” indicates that the units separated thereby are linked randomly or in any order;

R 1 is hydrogen, an aryl group, a heterocyclic group, a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group, or a C 1 -C 12 linear or branched alkyl group optionally substituted with one or more substituents;

each instance of R 2 is independently hydrogen or a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group;

R 3a and R 3a are each independently a methylene or ethylene group;

each instance of R 13 is independently hydrogen or a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group;

each instance of R 4 is —C(O)O—, —C(O) NH—, or —S(O)(O)—;

each instance of R 5 is independently an alkyl group, cycloalkyl group, alkenyl group, cycloalkenyl group, aryl group, heteroalkyl group, heterocyclic group, or combination thereof optionally comprising from 1 to 8 secondary or tertiary amines or a substituent comprising a tissue-specific or cell-specific targeting moiety; and

R 6 is hydrogen, an amino group, an aryl group, a heterocyclic group, a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group, a C 1 -C 12 linear or branched alkyl group optionally substituted with one or more amines; or a tissue-specific or cell-specific targeting moiety.

4. The polymer of claim 1 , wherein R 4 is —C(O)—O—.

5. The polymer of claim 1 , wherein R 5 is

wherein z is an integer from 1 to 5.

6. The polymer of claim 1 , wherein the ratio of (m 1 +n 1 )/(m 2 +n 2 ) is about 20 or less, and, optionally, about 0.2 or more.

7. The polymer of claim 1 , wherein the tissue-specific or cell-specific targeting moiety is:

wherein each of R 9 , R 10 , R 11 , and R 12 is independently hydrogen, halogen, C 1 -C 4 alkyl, or C 1 -C 4 alkoxy, optionally substituted with one or more amino groups; or an antibody or protein antigen.

8. The polymer of claim 1 , wherein the polymer is a cationic polymer.

9. A composition comprising e the polymer of claim 1 and a nucleic acid and/or polypeptide.

10. The composition of claim 9 , wherein the composition comprises a guide nucleic acid and/or donor nucleic acid; an RNA-guided endonuclease or nucleic acid encoding same; or combination thereof.

11. The composition of claim 10 , wherein the RNA-guided endonuclease is Cas9, Cpf1, or a combination thereof.

12. The composition of claim 9 , wherein the composition comprises a DNA recombinase, a zinc finger nuclease, a transcription activator-like effector nuclease, or a combination thereof.

13. The composition of claim 9 , wherein the composition comprises a nanoparticle comprising the polymer and the nucleic acid and/or polypeptide.

14. The composition of claim 9 , wherein the composition comprises a second polymer that comprises polyethylene oxide.

15. A method of delivering a nucleic acid and/or polypeptide to a cell, the method comprising administering the composition of claim 9 to the cell.

16. The method of claim 15 , wherein the cell is in a subject and the composition is administered to the subject, and wherein the polymer comprises a tissue-specific targeting moiety that localizes the polymer to tissues of a peripheral nervous system, a central nervous system, liver, muscle, lung, bone, or an eye of the subject.

17. The method of claim 16 , wherein the polymer comprises a targeting moiety that preferentially binds to tumor cells.

18. A polymer comprising a structure of Formula 1:

wherein:

each of m 1 and n 1 is an integer from 0 to 1000;

each of m 2 and n 2 is an integer from 0 to 1000, provided that the sum of m 2 +n 2 is greater than 5;

the symbol “/” indicates that the units separated thereby are linked randomly or in any order;

R 3a and R 3b are each independently a methylene or ethylene group;

each instance of R 13 is independently hydrogen or a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group;

A 1 and A 2 are each independently a group of formula

—(CH 2 ) p1 —[NR 2 —(CH 2 ) q1 -] r1 NR 2 2 ;

—(CH 2 ) p2 N[—(CH 2 ) q2 —NR 2 2 ] 2 ;

—(CH 2 ) p3 —N{[—(CH 2 ) q3 —NR 2 2 ][—(CH 2 ) q4 —NR 2 -]r 2 R 2 }; or

—(CH 2 ) p4 —N{—(CH 2 ) q5 —N[—(CH 2 ) q6 —NR 2 2 ] 2 } 2 ,

B 1 and B 2 are each independently

—(CH 2 ) p1 —[NR 2 —(CH 2 ) q1 -] r1 NR 2 —(CH 2 ) s1 —R 4 —R 5 ;

—(CH 2 ) p2 —N[—(CH 2 ) q2 —NR 2 —(CH 2 ) s2 —R 4 —R 5 ] 2 ;

—(CH 2 ) p3 —N{[—(CH 2 ) q3 —NR 2 2 ][—(CH 2 ) q4 —NR 2 —] r2 (CH 2 ) s3 —R 4 —R 5 };

—(CH 2 ) p4 —N{—(CH 2 ) q5 —N[—(CH 2 ) q6 —NR 2 —(CH 2 ) s4 —R 4 —R 5 ] 2 } 2 ;

—(CH 2 ) p1 —[NR 2 —(CH 2 ) q1 —] r1 NR 2 —CH 2 —CHOH—R 5 ;

—(CH 2 ) p2 —N[—(CH 2 ) q2 —NR 2 —CH 2 —CHOH—R 5 ] 2 ;

—(CH 2 ) p3 —N{[—(CH 2 ) q3 —NR 2 2 ][—(CH 2 ) q4 —NR 2 —] r2 —CH 2 —CHOH—R 5 };

—(CH 2 ) p4 —N{—(CH 2 ) q5 —N[—(CH 2 ) q6 —NR 2 —CH 2 —CHOH—R 5 ] 2 } 2 ;

—(CH 2 ) p2 —N[—(CH 2 ) q2 —NR 2 —(CH 2 ) s2 —R 5 ] 2 ;

—(CH 2 ) p3 —N{[—(CH 2 ) q3 —NR 2 2 ][—(CH 2 ) q4 —NR 2 —] r2 (CH 2 ) s3 —R 5 };

—(CH 2 ) p4 —N{—(CH 2 ) q5 —N[—(CH 2 ) q6 —NR 2 —(CH 2 ) s4 —R 5 | 2 } 2 ;

—(CH 2 ) p1 —[N{(CH 2 ) s1 —R 4 —R 5 }—(CH 2 ) q1 —] r1 NR 2 2 ; or

—(CH 2 ) p1 —[N{(CH 2 ) s1 —R 5 }—(CH 2 ) q1 -] r1 NR 2 2 ,

wherein p1 to p4, q1 to q6, r1 and r2, and s1 to s4 are each independently an integer of 1 to 5:

each instance of R 2 is independently hydrogen or a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group: each instance of R 4 is independently —C(O)O—, —C(O) NH—, or —S(O)(O)—; and each instance of R 5 is independently:

wherein

each instance of R 2 is independently hydrogen or a C 1 -C 12 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group;

R 7 is a C 1 -C 50 alkyl group, alkenyl group, cycloalkyl group, or cycloalkenyl group optionally substituted with one or more amines;

z is an integer from 1 to 5;

c is an integer from 0 to 50;

Y is optionally present and is a cleavable linker;

n is an integer from 0 to 50; and

R 8 is a tissue-specific or cell-specific targeting moiety.

19. A polymer of formula:

wherein (a+b) is from about 0 to about 75 and (c+d) is from about 5 to about 80.

20. A composition comprising the polymer of claim 19 and a nucleic acid and/or polypeptide.

21. The composition of claim 20 , wherein the composition comprises a guide nucleic acid and/or donor nucleic acid; an RNA-guided endonuclease or nucleic acid encoding same; or combination thereof.

22. The composition of claim 21 , wherein the RNA-guided endonuclease is Cas9, Cpf1, or a combination thereof.

23. The composition of claim 20 , wherein the composition comprises a DNA recombinase, a zinc finger nuclease, a transcription activator-like effector nuclease, or a combination thereof.

24. The composition of claim 20 , wherein the composition comprises a nanoparticle comprising the polymer and the nucleic acid and/or polypeptide.

25. The composition of claim 20 , wherein the composition comprises a second polymer that comprises polyethylene oxide.

26. A method of delivering a nucleic acid and/or polypeptide to a cell, the method comprising administering the composition of claim 20 to the cell.

27. The method of claim 26 , wherein the cell is in a subject and the composition is administered to the subject, and wherein the polymer comprises a tissue-specific targeting moiety that localizes the polymer to tissues of a peripheral nervous system, a central nervous system, liver, muscle, lung, bone, or an eye of the subject.

28. The method of claim 27 , wherein the polymer comprises a targeting moiety that preferentially binds to tumor cells.

Assignments (1)

Continuity (3)

References Cited (106)