# US 2020/0399424 A1 · Poly(amine-co-ester) polymers and polyplexes with modified end groups and methods of use thereof

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- Number: US 2020/0399424 A1
- Document ID: US20200399424A1
- Status: Published application
- Kind: A1
- Application number: 16/862,491
- Filed: 2020-04-29
- Published: 2020-12-24
- Earliest claimed: 2019-04-29
- Assignees: Yale University
- Applicants: Yale University
- Inventors: W. Mark Saltzman; Yuhang Jiang
- Web page: https://patentgrep.com/patent/US20200399424A1

## CPC

- C08G 63/685: containing nitrogen
- A61K 47/593: {Polyesters, e.g. PLGA or polylactide-co-glycolide}
- A61K 47/595: {Polyamides, e.g. nylon (polyamino acids A61K47/62)}
- A61K 48/0041: {the non-active part being polymeric}
- C08G 63/91: Polymers modified by chemical after-treatment
- C12N 15/88: using microencapsulation, e.g. using {amphiphile} liposome vesicle
- C08G: MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Poly(amine-co-ester) polymers, methods of forming active agent-load polyplexes and particles therefrom, and methods of using them for delivery of nucleic acid agents with optimal uptake have been developed. Examples demonstrate critical molecular weights in combination with exposed carboxylic and/or hydroxyl groups, and methods of making Typically, the compositions are less toxic, more efficient at drug delivery, or a combination thereof compared to a control other transfection reagents. In some embodiments, the compositions are suitable for in vivo delivery, and can be administered systemically to a subject to treat a disease or condition.

## Claims

### Claim 1

A polymer of Formula I:
wherein n is an integer from 1-30, m, o, and p are independently integers from 1-20, x, y, and q are independently integers from 1-1000, R is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl, or substituted or unsubstituted alkoxy, Z and Z′ are independently O or NR′, wherein R′ is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl,
wherein R1 and R2 are chemical entities containing a hydroxyl group, a primary amine group, a secondary amine group, a tertiary amine group, or combinations thereof.

### Claim 2 (depends on 1)

The polymer of claim 1, wherein R1 and/or R2 are not

### Claim 3 (depends on 1)

The polymer of claim 1, wherein polyplexes or particles formed from the polymer show improved loading, improved cellular transfection, improved intracellular endosomal release, or a combination thereof of a mRNA, relative to corresponding polyplexes or particles wherein R1 and/or R2 consist of

### Claim 4 (depends on 1)

The polymer of claim 1, wherein the polymer has a structure of Formula II:
wherein J1 and J2 are independently linking moieties or absent,
R3 and R4 are substituted alkyl containing a hydroxyl group, a primary amine group, a secondary amine group, a tertiary amine group, or combinations thereof.

### Claim 5 (depends on 1)

The polymer of claim 1, wherein the polymer has a structure of Formula III:

### Claim 6 (depends on 1)

The polymer of claim 1, wherein Z is the same as Z′.

### Claim 7 (depends on 1)

The polymer of claim 1, wherein n is 4, 10, 13, or 14.

### Claim 8 (depends on 1)

The polymer of claim 1, wherein m is 5, 6, or 7.

### Claim 9 (depends on 1)

The polymer of claim 1, wherein Rx is substituted or unsubstituted alkyl.

### Claim 10 (depends on 1)

The polymer of claim 1, wherein the weight average molecular weight, as measured by gel permeation chromatography using narrow polydispersity polystyrene standards, is between about 2,000 Daltons and 20,000 Daltons, preferably between about 2,000 Daltons and about 10,000 Daltons, most preferably between about 2000 Daltons and about 7,000 Daltons.

### Claim 11 (depends on 4)

The polymer of claim 4, wherein R3 is the same as R4.

### Claim 12 (depends on 4)

The polymer of claim 4, wherein R3 and/or R4 are independently selected from the group consisting of

### Claim 13 (depends on 4)

The polymer of claim 4, wherein J1 is —O— or —NH, J2 is —C(O)NH— or —C(O)O— or a combination thereof.

### Claim 14 (depends on 4)

The polymer of claim 4, wherein R3, R4 or both contain a primary amine group, and optionally one or more secondary or tertiary amine groups.

### Claim 15 (depends on 4)

The polymer of claim 4, wherein R3, R4 or both contain a hydroxyl group, and optionally one or more amine groups.

### Claim 16 (depends on 4)

The polymer of claim 4, wherein R3, R4 or both contain a hydroxyl group and no amine group.

### Claim 17 (depends on 4)

The polymer of claim 4, wherein at least one of R3 and R4 does not contain a hydroxyl group.

### Claim 18 (depends on 4)

The polymer of claim 4, wherein R3, R4 or both are -unsubstituted C1-C10 alkylene-Aq-unsubstituted C1-C10 alkylene-Bq, -unsubstituted C1-C10 alkylene-Aq-substituted C1-C10 alkylene-Bq, -substituted C1-C10 alkylene-Aq-unsubstituted C1-C10 alkylene-Bq, or -substituted C1-C10 alkylene-Aq-substituted C1-C10 alkylene-Bq, wherein Aq is absent or —NR5—, and Bq is hydroxyl, primary amine, secondary amine, or tertiary amine, wherein R5 is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl.

### Claim 19 (depends on 1)

A polyplex or solid-core particle comprising the polymer of claim 1, and one or more therapeutic, prophylactic, or diagnostic nucleic acid agents.

### Claim 20 (depends on 19)

The polyplex or particle of claim 19, wherein nucleic acid agent is RNA or DNA.

### Claim 21 (depends on 20)

The polyplex or particle of claim 20 wherein the nucleic acid agent comprises a coding sequence that encodes a protein.

### Claim 22 (depends on 21)

The polyplex or particle of claim 21 wherein the coding sequence is operably linked to an expression control sequence.

### Claim 23 (depends on 20)

The polyplex or particle of claim 20 wherein the nucleic acid agent is a functional nucleic acid, or an expression vector comprising sequence encoding a functional nucleic acid operably linked to an expression control sequence.

### Claim 24 (depends on 19)

The polyplex or particle of claim 19 wherein the nucleic acid agent is selected from the group consisting of antisense molecules, siRNA, miRNA, aptamers, ribozymes, triplex forming molecules, RNAi, and external guide sequences.

### Claim 25 (depends on 19)

A method of administering one or more therapeutic, diagnostic, or prophylactic nucleic acid agents, or combinations thereof in vivo, comprising administering the composition of claim 19.

### Claim 26 (depends on 25)

The method of claim 25 comprising transfecting cells by contacting cells with the particles or polyplex.

### Claim 27 (depends on 1)

A method of making the polymer of claim 1, comprising modifying the end groups of the unmodified polymer of Formula VII.
Wherein X′ is —OH or —NHR′.

### Claim 28 (depends on 5)

A method of making the polymer of claim 5, comprising (1) reacting the polymer of Formula VII with 1,1′-carbonyldiimidazole to obtain a polymer of Formula IV, and (2) reacting the polymer of Formula IV with R3-NH2 and R4-NH2.

### Claim 29 (depends on 28)

The method of claim 28, wherein R3, R4, or both are selected from the group below:
are the same.

### Claim 30

A method of increasing the transfection efficiency of a nucleic acid agent delivered using a polymer of Formula I:
wherein n is an integer from 1-30, m, o, and p are independently integers from 1-20, x, y, and q are independently integers from 1-1000, R is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl, or substituted or unsubstituted alkoxy, Z and Z′ are independently O or NR′, wherein R′ is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl,
wherein R1 and R2 are chemical entities containing a hydroxyl group, a primary amine group, a secondary amine group, a tertiary amine group, or combinations thereof, comprising
producing the polymer with a weight average molecular weight, as measured by gel permeation chromatography using narrow polydispersity polystyrene standards, is between about 2,000 Daltons and 20,000 Daltons, preferably between about 2,000 Daltons and about 10,000 Daltons, most preferably between about 2000 Daltons and about 7,000 Daltons, and
exposing hydroxyl and carboxyl end groups on the polymer.

### Claim 31

A polymer of Formula I:
wherein n is an integer from 1-30, m, o, and p are independently integers from 1-20, x, y, and q are independently integers from 1-1000, R is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl, or substituted or unsubstituted alkoxy, Z and Z′ are independently O or NR′, wherein R′ is hydrogen, substituted or unsubstituted alkyl, or substituted or unsubstituted aryl,
wherein R1 and R2 are chemical entities containing a hydroxyl group, a primary amine group, a secondary amine group, a tertiary amine group, or combinations thereof, comprising
having a weight average molecular weight, as measured by gel permeation chromatography using narrow polydispersity polystyrene standards, is between about 2,000 Daltons and 20,000 Daltons, preferably between about 2,000 Daltons and about 10,000 Daltons, most preferably between about 2000 Daltons and about 7,000 Daltons,
with exposed hydroxyl and carboxyl end groups on the polymer.

### Claim 32 (depends on 31)

The polymer of claim 31 complexed with a therapeutic, prophylactic or diagnostic agent.