Hybrid carriers for nucleic acid cargo

Patent metadata

Field	Value
Publication number	US-11478552-B2
Application number	US-201716308632-A
Country	US
Kind code	B2
Filing date	Jun 9, 2017
Priority date	Jun 9, 2016
Publication date	Oct 25, 2022
Grant date	Oct 25, 2022

Abstract

A composition for the delivery of a nucleic acid compound is provided which comprises a cationic peptide or polymer and a lipidoid compound. The nucleic acid compound may be any chemically modified or unmodified DNA or RNA. The amount of the lipidoid in the composition is preferably low, relative to the cationic peptide or polymer.

First claim

The invention claimed is:

1. A method of prophylaxis, treatment, and/or amelioration of a disease or disorder in a patient in need thereof comprising administering to the patient a safe and effective amount of a composition comprising a plurality of nanoparticles, wherein each nanoparticle of the plurality of nanoparticles comprises:
(a) a cationic peptide or cationic polymer;
(b) a cationic or permanent cationic lipidoid compound; and
(c) a nucleic acid compound;

wherein the cationic or permanent cationic lipidoid compound comprises two or three moieties of formula IIa and/or formula IIb:
—N(R 1 )—CH 2 —CH(R 5 )—R 2   (formula IIa)

—N + (R 3 )(R 4 )—CH 2 —CH(R 5 )—R 2   (formula IIb)

wherein independently for each individual moiety of formula IIa or formula IIb
R 1 is selected from hydrogen or C 1 -C 4 -alkyl,
R 2 is selected from linear or branched, saturated or unsaturated C 6 -C 16 hydrocarbyl chain,
R 3 and R 4 are selected from C 1 -C 4 -alkyl, and
R 5 is selected from hydrogen or hydroxyl; and

wherein the cationic peptide or cationic polymer is a compound according to formula
L 1 -P 1 -[P-] n -P 3 -L 2   (formula IV)

wherein
P is a cationic moiety having at least one —SH group capable of forming a disulfide linkage, or a disulfide-linked multimer thereof, wherein moiety P is selected from
a polymer moiety having a molecular weight from about 0.5 kDa to about 30 kDa, or
a peptide moiety composed of 3 to 100 amino acids, wherein at least 10% of the total number of amino acids of the peptide moiety represent basic amino acids selected from Arg, Lys, His and/or Orn;

P 3 is optional;
P 1 and P 3 are independently selected, each representing a linear or branched hydrophilic polymer chain selected from the group consisting of polyethylene glycol (PEG), poly-N-(2-hydroxypropyl)methacrylamide, poly-2-(methacryloyloxy)ethyl phosphorylcholines, poly(hydroxyalkyl L-asparagine), poly(2-(methacryloyloxy)ethyl phosphorylcholine), hydroxyethylstarch and poly(hydroxyalkyl L-glutamine), wherein the polymer chain exhibits a molecular weight from about 1 kDa to about 100 kDa, and wherein each of P 1 and P 3 is linked with a moiety P through a disulfide linkage;
L 1 and L 2 are optional ligands and independently selected from the group consisting of RGD, an RGD peptide, transferrin, folate, a signal peptide, a localization signal, a nuclear localization signal (NLS), an antibody, a cell penetrating peptide, a trans-activator of transcription (TAT), a ligand of a receptor, a cytokine, a hormone, a growth factor, a carbohydrate, a mannose, a galactose, an N-acetylgalactosamine, a synthetic ligand, an inhibitor of a receptor, an antagonist of a receptor, and a RGD peptidomimetic analogue;
n is an integer selected from 1 to about 50; and wherein, if n is greater than 1, each moiety P is linked with another moiety P through a disulfide linkage.

2. The method of claim 1 , wherein the disease is selected from the group consisting of cancer diseases, tumour diseases, infectious diseases, genetic diseases, and autoimmune diseases.

3. The method of claim 2 , wherein the disease is a viral, bacterial, or protozoological infectious disease.

4. The method of claim 1 , wherein the composition is administered via ocular delivery.

5. The method of claim 4 , wherein the ocular delivery is intravitreal, intracameral, subconjunctival, subretinal, subtenon, retrobulbar, topical, posterior juxtascleral administration or into the ciliary muscle.

6. The method of claim 5 , wherein the ocular delivery is into the ciliary muscle.

7. The method of claim 5 , wherein the plurality of nanoparticles comprises an mRNA encoding a protein such that the administration of the plurality of nanoparticles results in expression and/or activity of the protein encoded by the mRNA in the eye.

8. The method of claim 1 , wherein moiety P is
the peptide moiety composed of 7 to 30 amino acids, and wherein the at least one —SH group is provided by a Cys residue; or
the polymer moiety selected from an optionally modified polyacrylate, chitosan, polyethylenimine, polyamine, polyaminoesters, polyamidoamine, or a copolymer thereof.

9. The method of claim 8 , wherein the peptide moiety has two terminal ends, and wherein
the Cys residue is located at, or in proximity to, one of the terminal ends; or
the peptide moiety comprises at least two Cys residues, and wherein at least one of the Cys residues is located at, or in proximity to, each of the terminal ends.

10. The method of claim 1 , wherein the weight ratio of the cationic peptide or cationic polymer to the nucleic acid compound is at least about 1, and wherein the ratio of the cationic or permanent cationic lipidoid compound to the nucleic acid compound is not higher than about 15 nmol of the cationic or permanent cationic lipidoid compound per μg of the nucleic acid compound.

11. The method of claim 1 , wherein the weight ratio of the cationic or permanent cationic lipidoid compound to the cationic peptide or cationic polymer is not higher than about 1:50, and/or wherein the ratio of the cationic or permanent cationic lipidoid compound to the cationic peptide or cationic polymer is not higher than about 2 nmol of the cationic or permanent cationic lipidoid compound per μg of the cationic peptide or cationic polymer.

12. The method of claim 1 , wherein the N/P ratio of the basic groups of the cationic peptide or cationic polymer to the phosphate groups of the nucleic acid compound from about 0.1 to about 20, or from about 0.2 to about 15, or from about 2 to about 15, or from about 2 to about 12, wherein the N/P ratio is defined as the mole ratio of the nitrogen atoms of the basic groups of the cationic peptide or cationic polymer to the phosphate groups of the nucleic acid compound.

13. The method of claim 1 , wherein the plurality of nanoparticles comprises two or more different species of the cationic peptide and/or cationic polymer.

14. The method of claim 1 , wherein the plurality of nanoparticles further comprises one or more compounds independently selected from targeting agents, cell penetrating agents, and stealth agents.

15. The method of claim 1 , wherein the cationic or permanent cationic lipidoid compound is

optionally further comprising a pharmaceutically acceptable anion.

16. The method of claim 1 , wherein the nucleic acid compound is selected from the group consisting of chemically modified DNA, unmodified DNA, single stranded DNA, double stranded DNA, coding DNA, non-coding DNA, chemically modified RNA, unmodified RNA, single-stranded RNA, double-stranded RNA, coding RNA, and non-coding RNA.

17. The method of claim 16 , wherein the nucleic acid compound comprises a plasmid, an oligodesoxynucleotide, genomic DNA, a DNA primer, a DNA probe, an immunostimulatory DNA, an aptamer, a messenger RNA (mRNA), an oligoribonucleotide, a viral RNA, a replicon RNA, a transfer RNA (tRNA), a ribosomal RNA (rRNA), an immunostimulatory RNA (isRNA), a microRNA, a small interfering RNA (siRNA), a small nuclear RNA (snRNA), a small-hairpin RNA (shRNA), a riboswitch, an RNA aptamer, an RNA decoy, an antisense RNA, a ribozyme, or a combination thereof.

18. The method of claim 16 , wherein the nucleic acid compound comprises RNA.

19. The method of claim 18 , wherein the nucleic acid compound comprises mRNA.

Assignees

• Curevac Ag

Inventors

• Baumhof Patrick
• Thiele Carolin
• Rejman Joanna

Classifications

• C07C237/10

  having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by nitrogen atoms not being part of nitro or nitroso groups · CPC title

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title

• C07C233/20

  having the carbon atom of the carboxamide group bound to a carbon atom of an acyclic unsaturated carbon skeleton · CPC title

• A61P17/00

  Drugs for dermatological disorders · CPC title

• A61P3/00

  Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00) · CPC title