Therapeutic muscular dystrophy drug having bubble liposome loaded with morpholino as active ingredient

• US 9,457,102 B2
• Filed: 04/25/2012
• Issued: 10/04/2016
• Est. Priority Date: 05/09/2011
• Status: Active Grant

• Current Assignee: Nepa Gene Co., Ltd.
• Original Assignee: Nepa Gene Co., Ltd.
• Inventors: Negishi, Yoichi, Takahashi, Yoko, Aramaki, Yukihiko, Maruyama, Kazuo
• Primary Examiner(s): Kelly, Robert M
• Application Number:
  US14/116,316
• Publication Number:
  US 20140155813A1
• Time in Patent Office: 1,623 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 41/0028 Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
• CPC Class Codes: A61K 47/24 containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
• CPC Class Codes: A61K 47/6911 the form being a liposome
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• 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 21/04 for myasthenia gravis
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

To provide a technology for introducing a PMO with remarkably enhanced cell permeability, leading to significantly enhanced introduction efficiency to thereby provide a therapeutic drug that dramatically ameliorates a medical condition of DMD. Also provided is a therapeutic drug for Duchenne muscular dystrophy, including as an active ingredient a Bubble liposome having a specific morpholino oligomer (PMO) bound to a surface thereof, in which the PMO is introduced into a muscle fiber (muscle cell) of a muscle tissue with high efficiency by administration of the therapeutic drug into the muscle tissue or into a blood vessel followed by ultrasound irradiation to the muscle tissue transcutaneously.

Claims

1. A method for treating Duchenne muscular dystrophy in a subject in need thereof, the method comprising:

administering a Bubble liposome and a morpholino oligomer bound to a surface of the Bubble liposome into a blood vessel of the subject, and

transcutaneously irradiating a muscle tissue with ultrasound,

wherein the morpholino oligomer in a splicing process for a pre-mRNA of a dystrophin gene having a stop codon resulting from a mutation, produces a mature mRNA skipping an exon having the mutation therein;

the morpholino oligomer comprises a base sequence complementary to a region comprising a splicing enhancer sequence for said exon having the mutation therein; and

the morpholino oligomer has a polymerization degree of from 15 to 50 mer.

2. The method according to claim 1,

wherein said exon having the mutation therein comprises any one of exons 2, 8, 23, 43 to 46, and 50 to 53.

3. The method according to claim 1,

wherein the Bubble liposome comprises a PEG-liposome encapsulating a perfluorohydrocarbon, and

the Bubble liposome has an average particle diameter of from 50 to 500 nm.

4. The method according to claim 1,

wherein the administering comprises systemic administration via the great saphenous vein.

5. The method according to claim 4,

wherein the muscle tissue comprises a heart muscle tissue, a diaphragm muscle tissue, or both.