# US 10,478,448 B2 · Methods and compositions for modulating apolipoprotein (a) expression

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,478,448 B2
- Document ID: US10478448B2
- Status: Granted
- Kind: B2
- Application number: 15/855,203
- Filed: 2017-12-27
- Published: 2018-09-13
- Granted: 2019-11-19
- Earliest claimed: 2012-05-17
- Assignees: Ionis Pharmaceuticals, Inc.
- Applicants: Ionis Pharmaceuticals, Inc.
- Inventors: Rosanne M. Crooke; Mark J. Graham; Susan M. Freier; Marc Lim; Andrew Dibble
- Web page: https://patentgrep.com/patent/US10478448B2

## CPC

- A61K 31/7115: Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
- A61K 31/712: Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
- A61K 31/7125: Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters
- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- 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)}
- C12N 2310/11: Antisense
- C12N 2310/315: Phosphorothioates
- C12N 2310/321: 2'-O-R Modification
- C12N 2310/323: modified ring structure
- C12N 2310/3231: having an additional ring, e.g. LNA, ENA
- C12N 2310/3341: 5-Methylcytosine
- C12N 2310/3525: MOE, methoxyethoxy
- C12N 2320/50: Methods for regulating/modulating their activity
- C12N 2320/51: modulating the chemical stability, e.g. nuclease-resistance
- C12N 2320/52: modulating the physical stability, e.g. GC-content
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Disclosed herein are antisense compounds and methods for decreasing apo(a) to treat, prevent, or ameliorate diseases, disorders or conditions related to apo(a) or Lp(a). Certain diseases, disorders or conditions related to apo(a) or Lp(a) include inflammatory, cardiovascular and/or metabolic diseases, disorders or conditions. The antisense compounds disclosed herein can be used to treat such diseases, disorders or conditions in an individual in need thereof.

## Claims

### Claim 1

A method of treating, slowing the progression of, or ameliorating a symptom of hyperlipidemia in a subject, comprising:
administering to said subject a compound comprising a modified oligonucleotide, wherein:
the modified oligonucleotide consists of 14 to 25 linked nucleosides and comprises a nucleobase sequence comprising a portion of at least 14 contiguous nucleobases complementary to an equal length portion of nucleobases 3900 to 3923 of SEQ ID NO: 1, wherein the nucleobase sequence of the modified oligonucleotide is at least 95% complementary to SEQ ID NO: 1, including any salt thereof, whereby the administration treats, slows the progression of, or ameliorates a symptom of hyperlipidemia in the subject.

### Claim 2 (depends on 1)

The method of claim 1, wherein the subject has elevated apo(a) and/or Lp(a) levels.

### Claim 3 (depends on 1)

The method of claim 1, wherein apo(a) mRNA and/or protein levels are lowered.

### Claim 4 (depends on 1)

The method of claim 1, wherein the modified oligonucleotide consists of 18 to 24, 19 to 22, 15 to 25, 16 or 20 linked nucleosides.

### Claim 5 (depends on 1)

The method of claim 1, wherein the modified oligonucleotide comprises a nucleobase sequence comprising a portion of at least 16, at least 18, at least 19, or at least 20 contiguous nucleobases complementary to an equal length portion of SEQ ID NO: 1.

### Claim 6 (depends on 1)

The method of claim 1, wherein the nucleobase sequence of the modified oligonucleotide is at least 96%, 97%, 98% or 99% complementary to SEQ ID NO: 1, or is 100% complementary to SEQ ID NO:1.

### Claim 7 (depends on 1)

The method of claim 1, wherein the modified oligonucleotide consists of 14 to 25 linked nucleosides and comprises a nucleobase sequence comprising at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of the nucleobase sequence of SEQ ID NO: 58.

### Claim 8 (depends on 1)

The method of claim 1, wherein the modified oligonucleotide is single-stranded.

### Claim 9 (depends on 1)

The method of claim 1, wherein at least one internucleoside linkage is a modified internucleoside linkage.

### Claim 10 (depends on 9)

The method of claim 9, wherein at least one internucleoside linkage is a phosphorothioate internucleoside linkage.

### Claim 11 (depends on 1)

The method of claim 1, wherein the modified oligonucleotide comprises at least one modified sugar.

### Claim 12 (depends on 11)

The method of claim 11, wherein at least one modified sugar is a bicyclic sugar.

### Claim 13 (depends on 11)

The method of claim 11, wherein at least one modified sugar comprises a 2′-O-methoxyethyl, a constrained ethyl, a 3′-fluoro-HNA or a 4′- (CH2)n—O-2′ bridge, wherein n is 1 or 2.

### Claim 14 (depends on 1)

The method of claim 1, wherein at least one nucleoside comprises a modified nucleobase.

### Claim 15 (depends on 14)

The method of claim 14, wherein the modified nucleobase is a 5-methylcytosine.

### Claim 16 (depends on 1)

The method of claim 1, wherein the modified oligonucleotide consists of 14 to 25 linked nucleosides and comprises:
a gap segment consisting of linked deoxynucleosides;
a 5′ wing segment consisting of linked nucleosides;
a 3′ wing segment consisting of linked nucleosides;
wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

### Claim 17 (depends on 16)

The method of claim 16, wherein the modified oligonucleotide consists of 20 linked nucleosides and comprises:
a gap segment consisting of ten linked deoxynucleosides;
a 5′ wing segment consisting of five linked nucleosides;
a 3′ wing segment consisting of five linked nucleosides;
wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethyl sugar, wherein at least one internucleoside linkage is a phosphorothioate linkage and wherein each cytosine residue is a 5-methylcytosine.

### Claim 18 (depends on 16)

The method of claim 16, wherein the modified oligonucleotide consists of 20 linked nucleosides.

### Claim 19 (depends on 1)

The method of claim 1, wherein the modified oligonucleotide consists of 20 contiguous linked nucleosides of SEQ ID NO: 58 and the modified oligonucleotide further comprises:
a gap segment consisting of ten linked deoxynucleosides;
a 5′ wing segment consisting of five linked nucleosides;
a 3′ wing segment consisting of five linked nucleosides;
wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a 2′-O-methoxyethy sugar, wherein at least one internucleoside linkage is a modified internucleoside linkage and wherein each cytosine residue is a 5-methylcytosine.

### Claim 20 (depends on 1)

The method of claim 1, wherein a therapeutically effective dose of the modified oligonucleotide is administered, whereby hyperlipidemia in the subject is treated, the progression slowed or a symptom ameliorated.

### Claim 21 (depends on 20)

The method of claim 20, wherein the subject has or has had aneurysm, angina, arrhythmia, atherosclerosis, cerebrovascular disease, coronary artery disease, coronary heart disease, myocardial infarction, peripheral vascular disease, peripheral artery disease, peripheral artery occlusive disease, retinal vascular occlusion, or stroke.

### Claim 22 (depends on 1)

The method of claim 1, wherein the subject is a human subject.

### Claim 23 (depends on 1)

The method of claim 1, wherein the compound is a first agent and is co-administered with one or more secondary agents or therapy.

### Claim 24 (depends on 1)

The method of claim 1, wherein the modified oligonucleotide has a viscosity of less than 40 cP.

### Claim 25 (depends on 23)

The method of claim 23, wherein the one or more secondary agents or therapy is/are selected from among an apo(a) lowering agent, a Lp(a) lowering agent, an agent to reduce thromboembolism formation, a glucose-lowering agent, a LDL lowering agent, a triglyceride lowering agent, a cholesterol lowering agent, a HDL raising agent, an anti-inflammatory agent, an Alzheimer Disease drug, a non-steroidal anti-inflammatory drug (NSAID), fish oil, niacin, nicotinic acid, an apoB inhibitor, a CETP inhibitor, a thyroid hormone analog, a HMG-CoA reductase inhibitor, a fibrate, a microsomal triglyceride transfer protein inhibitor and Lp(a) apheresis.