# US 9,872,844 B2 · Compositions and methods for the reduction or prevention of hepatic steatosis

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- Number: US 9,872,844 B2
- Document ID: US9872844B2
- Status: Granted
- Kind: B2
- Application number: 14/843,873
- Filed: 2015-09-02
- Published: 2016-03-10
- Granted: 2018-01-23
- Earliest claimed: 2014-02-27
- Assignees: NuSirt Sciences, Inc.
- Applicants: Nusirt Sciences, Inc.
- Inventors: Michael Zemel; Antje Bruckbauer
- Web page: https://patentgrep.com/patent/US9872844B2

## CPC

- A61K 31/198: Alpha-amino acids, e.g. alanine or edetic acid [EDTA] (betaine A61K31/205; proline A61K31/401; tryptophan A61K31/405; histidine A61K31/4172; peptides not degraded to individual amino acids A61K38/00)
- A61K 31/05: Phenols {(cannabinoids A61K31/658)}
- A61K 31/155: Amidines ([IMAGE cpc-sch-A61K-1029.gif]), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
- A61K 31/4439: containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole (nicotine A61K31/465)
- A61K 31/4985: Pyrazines or piperazines ortho- or peri-condensed with heterocyclic ring systems
- A61K 31/519: ortho- or peri-condensed with heterocyclic rings
- A61K 31/522: having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
- A61K 31/53: having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine (melarsoprol A61K31/555 {; with four nitrogen atoms A61K31/495})
- A61K 31/7048: having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin {, digitoxin or digoxin}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 3/04: Anorexiants; Antiobesity agents
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- 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

## Abstract

Methods useful for reducing or preventing non-alcoholic steatohepatitis or hepatic steatosis are provided herein. Such methods may comprise administering to a subject in need thereof a sirtuin pathway activator and/or PDE5 inhibitor alone or in combination with an amount of a branched amino acid in free amino acid form, or a metabolite thereof. Also provided herein are compositions and kits for practicing any of the methods described herein.

## Claims

### Claim 1

A method of reducing non-alcoholic steatohepatitis (NASH) in a subject in need thereof, comprising administering to the subject a composition comprising:
a) an amount of leucine in the form of a free amino acid and/or metabolite thereof;
b) an amount of metformin; and
c) an amount of sildenafil;
wherein the amount of component (a) administered to the subject is about 0.25-3 g/day of leucine in the form of a free amino acid, and/or about 0.2-3 g/day of leucine metabolite;
wherein the amount of component (b) administered to the subject is about 10-1000 mg/day of metformin;
wherein the amount of component (c) administered to the subject is about 0.05-100 mg/day of sildenafil; and
wherein components (a), (b), and (c) are co-administered to the subject simultaneously as a single composition or sequentially.

### Claim 2 (depends on 1)

The method of claim 1, wherein the leucine metabolite thereof is β-hydroxy β-methylbutyrate (HMB) or keto-isocaproic acid (KIC).

### Claim 3 (depends on 1)

The method of claim 1, wherein the molar ratio of leucine to sildenafil in the composition is at least about 20.

### Claim 4 (depends on 1)

The method of claim 1, wherein a sub-therapeutic amount of metformin is administered to the subject.

### Claim 5 (depends on 1)

The method of claim 1, wherein component (a) comprises leucine in the form of a free amino acid, component (b) comprises metformin, and component (c) comprises sildenafil.

### Claim 6 (depends on 5)

The method of claim 5, wherein the composition administered to the subject is formulated in a unit dose, and wherein the composition comprises:
component (a) comprising about 900-1200 mg of leucine in the form of a free amino acid;
(ii) component (b) comprising about 0.1-10 mg of sildenafil; and
(iii) component (c) comprising about 200-550 mg of metformin.

### Claim 7 (depends on 6)

The method of claim 6, wherein the unit dose is administered twice a day.

### Claim 8 (depends on 1)

The method of claim 1, wherein the subject has been diagnosed with the non-alcoholic steatohepatitis (NASH), or exhibits a non-alcoholic fatty liver disease (NAFLD) selected from the group consisting of non-alcoholic fatty liver (NAFL), NASH-related cirrhosis and hepatic steatosis.

### Claim 9 (depends on 1)

The method of claim 1, comprising administering the composition to the subject twice daily and the liver mass of the subject is decreased by 25% within 6 weeks.

### Claim 10 (depends on 1)

The method of claim 1, wherein the composition is substantially free of one or more free amino acids selected from the group consisting of alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

### Claim 11 (depends on 1)

The method of claim 1, wherein the administering of (a), (b), and (c) reduces non-alcoholic steatohepatitis (NASH) to a greater extent than administration of (a) alone, thereby reducing non-alcoholic steatohepatitis (NASH) in the subject.

### Claim 12 (depends on 11)

The method of claim 11, wherein the non-alcoholic steatohepatitis (NASH) in the subject is evidenced by an accumulation of hepatic fat, fibrosis and/or inflammation detectable by one or more methods selected from the group consisting of ultrasonography, computed tomography (CT), magnetic resonance imaging, magnetic resonance spectroscopy (MRS), magnetic resonance elastography (MRE), transient elastography (TE), hepatic steatosis, inflammation and/or fibrosis detected by biopsy, and detection of a biomarker indicative of liver damage.

### Claim 13 (depends on 12)

The method of claim 12, wherein the biomarker indicative of liver damage is elevated serum alanine aminotransferase, aspartate aminotransferase, and/or cytokeratin 18.

### Claim 14

A method of treating non-alcoholic steatohepatitis (NASH) in a subject in need thereof, comprising administering to the subject a composition comprising:
a. an amount of leucine in the form of a free amino acid and/or metabolite thereof;
b. an amount of metformin; and
c. an amount of sildenafil;
wherein the amount of component (a) administered to the subject is about 0.25-3 g/day of leucine in the form of a free amino acid, and/or about 0.2-3 g/day of leucine metabolite;
wherein the amount of component (b) administered to the subject is about 10-1000 mg/day of metformin;
wherein the amount of component (c) administered to the subject is about 0.05-100 mg/day of sildenafil; and
wherein components (a), (b), and (c) are co-administered to the subject simultaneously as a single composition or sequentially.

### Claim 15 (depends on 14)

The method of claim 14, wherein the composition administered to the subject is formulated in a unit dose, and wherein the composition comprises:
component (a) comprising about 900-1200 mg of leucine in the form of a free amino acid; and
(ii) component (b) comprising about 200-550 mg of metformin and about 0.1-10 mg of sildenafil.

### Claim 16 (depends on 15)

The method of claim 15, wherein the components of the unit dose are co-administered to the subject simultaneously as a single composition or sequentially, and the unit dose is administered twice a day.

### Claim 17 (depends on 1, 14)

The method of claim 1 or 14, wherein the composition further comprises a sirtuin pathway activator.

### Claim 18 (depends on 17)

The method of claim 17, wherein the sirtuin pathway activator comprises an AMPK activator, a PGC-1α activator, a PDE5 specific inhibitor, a biguanide or a thiazolidinedione.

### Claim 19 (depends on 17)

The method of claim 17, wherein the thiazolidinedione comprises rosiglitazone or pioglitazone.

### Claim 20 (depends on 17)

The method of claim 17, wherein the amount of thiazolidinedione administered to the subject is about 0.1-4 mg of rosiglitazone or about 0.1-45 mg of pioglitazone.

### Claim 21 (depends on 18)

The method of claim 18, wherein the PDE5 specific inhibitor is selected from the group consisting of icariin, tadalafil, vardenafil, avanafil, lodenafil, mirodenafil and udenafil.

### Claim 22 (depends on 18)

The method of claim 18, wherein the amount of the PDE5 specific inhibitor is selected from the group consisting of about 1-2000 mg/day of icariin, about 0.01-20 mg/day of tadalafil, about 0.01-20 mg/day of vardenafil, about 1-200 mg/day of avanafil, about 1-200 mg/day of lodenafil, about 1-100 mg/day of mirodenafil, about 1-200 mg/day of udenafil, and about 1-2000 mg/day of zaprinast.