Compositions and methods for delivery of agents

• US 10,556,018 B2
• Filed: 08/10/2017
• Issued: 02/11/2020
• Est. Priority Date: 12/10/2015
• Status: Active Grant

• Current Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Original Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Inventors: Besin, Gilles, Hoge, Stephen, Senn, Joseph, Benenato, Kerry, Sabnis, Staci
• Primary Examiner(s): Zara, Jane J
• Application Number:
  US15/674,107
• Publication Number:
  US 20180147298A1
• Time in Patent Office: 915 Days
• Field of Search: 514 44 R, 424 91, 435 61, 435 911, 435 9131, 435455, 435458, 536 231, 536 245
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 31/7115 Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61K 47/14 Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
• CPC Class Codes: A61K 47/18 Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
• CPC Class Codes: A61K 47/543 Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
• CPC Class Codes: A61K 47/544 Phospholipids
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/6929 the form being a nanoparticle, e.g. an immuno-nanoparticle
• CPC Class Codes: A61K 48/0033 the non-active part being non-polymeric
• CPC Class Codes: A61K 48/0066 Manipulation of the nucleic acid to modify its expression pattern, e.g. enhance its duration of expression, achieved by the presence of particular introns in the delivered nucleic acid
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: C08G 2650/04 : End-capping
• CPC Class Codes: C08G 65/3322 : acyclic
• CPC Class Codes: C08G 65/3326 : aromatic
• CPC Class Codes: C08G 65/33306 : acyclic
• CPC Class Codes: C08G 65/3331 : cyclic
• CPC Class Codes: C08G 65/33317 : heterocyclic
• CPC Class Codes: C08G 65/33324 : acyclic
• CPC Class Codes: C08G 65/3348 : containing nitrogen in addition to sulfur
• CPC Class Codes: C08G 65/3356 : having nitrogen in addition to phosphorus
• CPC Class Codes: C12N 15/117 : Nucleic acids having immunomodulatory properties, e.g. containing CpG-motifs
• CPC Class Codes: C12N 15/88 : using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12N 2310/141 : MicroRNAs, miRNAs
• CPC Class Codes: C12N 2310/17 : Immunomodulatory nucleic acids
• CPC Class Codes: C12N 2320/32 : Special delivery means, e.g. tissue-specific

Abstract

This disclosure provides improved lipid-based compositions, including lipid nanoparticle compositions, and methods of use thereof for delivering agents in vivo including nucleic acids and proteins. These compositions are not subject to accelerated blood clearance and they have an improved toxicity profile in vivo.

Claims

1. A method for delivering a therapeutic level of a protein of interest to a subject, the method comprising:

administering to the subject at least two doses of lipid nanoparticles (LNPs) comprising an mRNA coding for the protein of interest, wherein the LNPs comprise an ionizable lipid, a helper lipid, a structural lipid, and a PEG-lipid,

wherein the PEG-lipid is a compound of Formula (V—

OH);

2. The method of claim 1, wherein the compound of Formula (V—

OH) is Cmpd452;

3. The method of claim 2, wherein the PEG-lipid comprises a PEG molecule of an average molecular weight of 2,000 Da or less.

4. The method of claim 3, wherein the PEG-lipid is HO-PEG2000-ester-C18 (Cmpd403).

5. The method of claim 1, wherein the ionizable lipid is a compound of Formula (XI):

6. The method of claim 5, wherein R′

is C₉ alkyl.

7. The method of claim 5, wherein n is 2.

8. The method of claim 5, wherein the compound of Formula (XI) has the structure of Cmpd18:

9. The method of claim 1, wherein the helper lipid comprises at least one fatty acid chain of at least 8 carbons in length and at least one polar headgroup moiety, and wherein the helper lipid does not comprise a phosphatidyl choline (PC) and is a zwitterionic non-cationic helper lipid, a DSPC analog, oleic acid, an oleic acid analog, DSPC, or a 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) substitute.

10. The method of claim 9, wherein the helper lipid is DSPC.

11. The method of claim 1, wherein the helper lipid is oleic acid or an oleic acid analog.

12. The method of claim 1, wherein the LNPs comprise a molar ratio of about 45-65% ionizable lipid:

5-25% helper lipid;

15-45% structural lipid; and

0.15-15% PEG-lipid.

13. The method of claim 12, wherein the PEG-lipid is present in the LNPs in an amount of less than 0.5% (w/w).

14. The method of claim 1, wherein said administering results in a reduced IgG response in the subject.

15. The method of claim 1, wherein said administering results in a reduced anti-DSPC IgM response in the subject.

16. The method of claim 1, wherein said administering results in reduced B1a cell activation in the subject.

17. The method of claim 1, wherein said administering results in reduced CD36 positive cell activation in the subject.

18. The method of claim 1, wherein said at least two doses of LNPs is more than two doses of LNPs.

19. The method of claim 1, wherein the interval between the doses is about 21 days or less.

20. The method of claim 1, wherein the mRNA is chemically-modified.

21. The method of claim 1, wherein the mRNA comprises at least one miR binding site.

22. The method of claim 21, wherein the miR binding site is selected from miR 122, 126, miR 155, and miR 142.3p.

23. The method of claim 1, further comprising administering to the subject an additional agent that inhibits immune responses.

24. The method of claim 1, comprising:

administering to the subject a first dose of lipid nanoparticles (LNPs) comprising an mRNA coding for the protein of interest, wherein the LNPs comprise an ionizable lipid, a helper lipid, a structural lipid, and a PEG-lipid, wherein;

the PEG-lipid

25. The method of claim 24, wherein the PEG-lipid is HO-PEG2000-ester-C18(Cmpd403).

26. The method of claim 24, wherein the compound of Formula (XI) has the structure of Cmpd18:

27. The method of claim 24, wherein the helper lipid is DSPC.

28. The method of claim 24, wherein the structural lipid is cholesterol.