# US 10,765,758 B2 · Optimized in vivo delivery system with endosomolytic agents for nucleic acid conjugates

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- Number: US 10,765,758 B2
- Document ID: US10765758B2
- Status: Granted
- Kind: B2
- Application number: 16/458,214
- Filed: 2019-07-01
- Published: 2019-10-17
- Granted: 2020-09-08
- Earliest claimed: 2010-06-22
- Assignees: Onxeo; Institut Curie; Centre National De La Recherche Scientifique
- Applicants: Onxeo; Institut Curie; Centre National De La Recherche Scientifique
- Inventors: Jian-Sheng Sun; Marie Dutreix; Maria Quanz
- Web page: https://patentgrep.com/patent/US10765758B2

## CPC

- A61K 47/549: {Sugars, nucleosides, nucleotides or nucleic acids}
- A61K 31/4706: 4-Aminoquinolines; 8-Aminoquinolines, e.g. chloroquine, primaquine
- A61K 31/7088: Compounds having three or more nucleosides or nucleotides
- A61K 31/713: Double-stranded nucleic acids or oligonucleotides
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61K 47/542: {Carboxylic acids, e.g. a fatty acid or an amino acid}
- A61K 47/543: {Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine}
- A61K 47/551: {one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid}
- A61K 47/554: {the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid}
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61P 35/00: Antineoplastic agents
- C07H 21/04: with deoxyribosyl as saccharide radical
- C12N 15/00: Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor (mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00)
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates to an optimized in vivo delivery system with endosomolytic agents for nucleic acid of therapeutic interest conjugated to molecules facilitating endocytosis, in particular for use in the treatment of cancer.

## Claims

### Claim 1

A conjugated hairpin nucleic acid molecule which has the following formula:
wherein N is a nucleotide, n is an integer from 15 to 195, the underlined N refers to a nucleotide having or not a modified phosphodiester backbone, L′ is a linker selected from the group consisting of an abasic nucleotide, a polyether, a polyamine, a polyamide, a peptide, a carbohydrate, a lipid, a polyhydrocarbon, an oligoethylene glycol, hexaethyleneglycol, tetradeoxythymidylate (T4), 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane and 2,19-bis(phosphor)-8-hydraza-1-hydroxy-4-oxa-9-oxo-nonadecane; m is 1 and L is a linker, C is a molecule facilitating endocytosis selected from the group consisting of single or double chain fatty acids, tocopherol, folates or folic acid, cholesterol, sugar, peptide and protein, and wherein C is conjugated via L to the linker L′.

### Claim 2 (depends on 1)

The conjugated nucleic acid molecule according to claim 1, wherein n is 27.

### Claim 3 (depends on 1)

The conjugated nucleic acid molecule according to claim 1, wherein C is selected from the group consisting of tocopherol and cholesterol.

### Claim 4 (depends on 1)

The conjugated nucleic acid molecule according to claim 1, wherein L′ is a loop linking the two strands of the nucleic acid molecule.

### Claim 5 (depends on 1)

The conjugated nucleic acid molecule according to, claim 1 wherein L′ is a non-nucleotide linker selected from the group consisting of abasic nucleotide, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon and an oligoethylene glycol.

### Claim 6 (depends on 1)

The conjugated nucleic acid molecule according to claim 1, wherein L′ is selected from the group consisting of hexaethyleneglycol, tetradeoxythymidylate (T4), 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane and 2,19-bis(phosphor)-8-hydraza-1-hydroxy-4-oxa-9-oxo-nonadecane.

### Claim 7 (depends on 1)

The conjugated nucleic acid molecule according to claim 1, wherein L is selected from the group consisting of aliphatic chain, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon, an oligoethylene glycol, a dialkyl-disulfide, compounds incorporating a disulfide linkage, a protected disulfide linkage, an amide linkage, an acid labile linkage, an ester linkage, an ortho ester linkage, a phosphonamide linkage, a biocleavable peptide linkage, an azo linkage and an aldehyde linkage.

### Claim 8 (depends on 7)

The conjugated nucleic acid molecule according to claim 7, wherein L is selected from the group consisting of an oligoethylene glycol and a dialkyl-disulfide.

### Claim 9 (depends on 1)

A pharmaceutical composition comprising a conjugated nucleic acid molecule according to claim 1.

### Claim 10 (depends on 9)

The pharmaceutical composition according to claim 9, wherein said composition further comprises a DNA-damaging anti-tumoral agent and a pharmaceutically acceptable carrier.

### Claim 11 (depends on 1)

A kit comprising a conjugated nucleic acid molecule according to claim 1 and a DNA-damaging anti-tumoral agent, as a combined preparation for simultaneous, separate or sequential use.

### Claim 12 (depends on 1)

A method for treating a cancer in a subject in need thereof, comprising administering an effective amount of conjugated nucleic acid molecule of claim 1.

### Claim 13 (depends on 12)

The method according to claim 12, wherein the treatment further comprises radiotherapy or chemotherapy, optionally with a DNA damaging antitumoral agent.

### Claim 14 (depends on 1)

The conjugated nucleic acid molecule according to claim 1, wherein the molecule has the following formula

### Claim 15 (depends on 6)

The conjugated nucleic acid molecule according to claim 6, wherein L′ is 1,19-bis(phospho)-8-hydraza-2-hydroxy-4-oxa-9-oxo-nonadecane.

### Claim 16 (depends on 6)

The conjugated nucleic acid molecule according to claim 6, wherein L′ is 2,19-bis(phosphor)-8-hydraza-1-hydroxy-4-oxa-9-oxo-nonadecane.

### Claim 17 (depends on 14)

A pharmaceutical composition comprising a conjugated nucleic acid molecule according to claim 14.

### Claim 18 (depends on 14)

A method for treating a cancer in a subject in need thereof, comprising administering an effective amount of the conjugated nucleic acid molecule of claim 14 to said subject.