MRNA therapy for phenylketonuria

• US 9,522,176 B2
• Filed: 10/22/2014
• Issued: 12/20/2016
• Est. Priority Date: 10/22/2013
• Status: Active Grant

• Current Assignee: Translate Bio, Inc. (Sanofi )
• Original Assignee: Shire Human Genetic Therapies, Inc. (Shire plc)
• Inventors: DeRosa, Frank, Heartlein, Michael, Dias, Anusha
• Primary Examiner(s): LEAVITT, MARIA GOMEZ
• Application Number:
  US14/521,323
• Publication Number:
  US 20150110858A1
• Time in Patent Office: 790 Days
• Field of Search: 514/44.R, 424/450, 435/320.1, 536/23.2
• US Class Current: 1/1
• CPC Class Codes: A61K 38/44 Oxidoreductases (1)
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 48/0033 the non-active part being non-polymeric
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12Y 114/16001 Phenylalanine 4-monooxygenase (1.14.16.1)

Abstract

The present invention provides, among other things, methods of treating phenylketonuria (PKU), including administering to a subject in need of treatment a composition comprising an mRNA encoding phenylalanine hydroxylase (PAH) at an effective dose and an administration interval such that at least one symptom or feature of PKU is reduced in intensity, severity, or frequency or has delayed in onset. In some embodiments, the mRNA is encapsulated in a liposome comprising one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids.

Claims

1. A method of treating phenylketonuria (PKU), the method comprising administering systemically to a subject in need of treatment a composition comprising an mRNA encoding a phenylalanine hydroxylase (PAH),

wherein the mRNA comprises a coding nucleotide sequence consisting of SEQ ID NO:

3,

wherein the mRNA is encapsulated within a liposome with a size of less than 100 nm, and

wherein administration of the composition at an effective dose and at an administration interval results in increased PAH protein level in the liver and in reduced phenylalanine level in the serum as compared to the baseline of PAH and phenylalanine levels before the treatment.

2. The method of claim 1, wherein the liposome comprises a cationic lipid, a non-cationic lipid, a cholesterol-based lipid and a polyethylene glycol (PEG)-modified lipid.

3. The method of claim 2, wherein the cationic lipid is selected from the group consisting of 1,1′

-((2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)2-hydroxydodecyl)amino)ethyl)piperazin-1-yl)ethyl)azanediyl)bis(dodecan-2-ol) (C12-200), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3), N,N-dimethyl-2,3-bis((9Z,12Z)-octadeca-9,12-dien-1-yloxy)propan-1-amine (DLinDMA), 2-(2,2-di((9Z,12Z)-octadeca-9,12-dien-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylethanamine (DLinKC2DMA, [XTC2]), 3,6-bis(4-(bis(2-hydroxydodecyl)amino)butyl)piperazine-2,5-dione (cKK-E12), 10,13-dimethyl-17-(6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl 3-(1H-imidazol-5-yl)propanoate (ICE), (15Z,18Z)-N,N-dimethyl-6-((9Z,12Z)-octadeca-9,12-dien-1-yl)tetracosa-15,18-dien-1-amine (HGT5000), (4Z,15Z,18Z)-N,N-dimethyl-6-((9Z,12Z)-octadeca-9,12-dien-1-yl)tetracosa-4,15,18-trien-1-amine (HGT5001), N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), 1,2-dimyristyloxyproyl-3-dimethyl-hydroxyethyl ammonium bromide (DMRIE), dioleoyloxy-N-[2-sperminecarboxamido)ethyl]-N,N-dimethyl-1-propaniminiumtrifluoroacetate (DOSPA), dioctadecylamidoglycyl spermine (DOGS), 1,2-dioleoyl-3-dimethylammonium-propane (DODAP), N,N-dimethyl-(2,3-dioleyloxy)propylamine (DODMA) and N,N-dimethyl-(2,3-dimyristyloxy)propylamine (DMDMA), 1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), (2S)-2-(4-((10,13-dimethyl-17-(6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)butoxy)-N,N-dimethyl-3-((9Z,12Z)-octadeca-9,12-dien-1-yloxy)propan-1-amine (CLinDMA), 2-[5′

-(cholest-5-en-3[betal]-oxy)-3′

-oxapentoxy)-3-dimethyl-1-1(cis,cis-9′

,12′

-octadecadienoxy) propane (CpLinDMA), N,N-dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N′

-dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), (9Z,9′

Z,12Z,12′

Z)-3-(dimethylamino)propane-1,2-diyl bis(octadeca-9,12-dienoate) (DLinDAP), 1,2-dilinoleylcarbamyl-3-dimethylaminopropane (DLinCDAP), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), 2-((2,3-bis((9Z,12Z)-octadeca-9,12-dien-1-yloxy)propyl)disulfanyl)-N,N-dimethylethanamine (HGT4003), and combinations thereof.

4. The method of claim 3, wherein the cationic lipid comprises cKK-E12, which has the structure:

5. The method of claim 2, wherein the cholesterol-based lipid is cholesterol or PEGylated cholesterol.

6. The method of claim 2, wherein the cationic lipid constitutes about 30-50% of the liposome by molar ratio.

7. The method of claim 2, wherein the ratio of cationic lipid:

non-cationic lipid;

cholesterol lipid;

PEGylated lipid is approximately 40;

30;

25;

5 by molar ratio.

8. The method of claim 1, wherein the liposome comprises a combination selected from the group consisting of:

cKK-E12, 1,2-dioleyl-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol and 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene Glycol (DMG-PEG2K);

C12-200, DOPE, cholesterol and DMG-PEG2K;

HGT4003, DOPE, cholesterol and DMG-PEG2K;

or

ICE, DOPE, cholesterol and DMG-PEG2K.

9. The method of claim 1, wherein the mRNA is administered at the effective dose ranging from about 0.01-5.0 mg/kg body weight.

10. The method of claim 1, wherein the composition is administered once a week.

11. The method of claim 1, wherein the mRNA comprises a 5′

UTR sequence of SEQ ID NO;

4.

12. The method of claim 1, wherein the mRNA comprises a 3′

UTR sequence of SEQ ID NO;

5 or SEQ ID NO;

6.