# US 2021/0275569 A1 · Methods for safely reducing thrombopoietin

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- Number: US 2021/0275569 A1
- Document ID: US20210275569A1
- Status: Published application
- Kind: A1
- Application number: 17/260,804
- Filed: 2019-07-16
- Published: 2021-09-09
- Earliest claimed: 2018-07-17
- Assignees: Aronora, Inc.; Oregon Health & Science University; Ionis Pharmaceuticals, Inc.
- Applicants: Aronora, Inc.; Oregon Health & Science University; Ionis Pharmaceuticals, Inc.
- Inventors: Andras Gruber; Erik Tucker; Brett P. Monia; Alexey Revenko
- Web page: https://patentgrep.com/patent/US20210275569A1

## 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 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 7/02: Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
- A61P 35/00: Antineoplastic agents
- C12N 15/1136: {against growth factors, growth regulators, cytokines, lymphokines or hormones}
- C12N 2310/11: Antisense
- C12N 2310/315: Phosphorothioates
- C12N 2310/321: 2'-O-R Modification
- C12N 2310/3341: 5-Methylcytosine
- C12N 2310/341: Gapmers, i.e. of the type ===---===
- C12N 2320/11: for the determination of target sites, i.e. of active nucleic acids
- C12N 2320/30: Special therapeutic applications
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided herein are methods, compounds, and compositions for safely reducing thrombopoietin in a cell or an individual. Such methods, compounds, and compositions maintain platelet count within a safe hemostatic range. Such methods, compounds, and compositions are useful to safely treat, prevent, or ameliorate a disease that can benefit from platelet count reduction in an individual. Such methods, compounds, and compositions are useful for treating or preventing diseases in which thrombopoietin contributes to the presence or activation of platelets contributes to and promotes disease initiation or progression, and/or adversely affects disease outcome.

## Claims

### Claim 1

A method of safely treating, preventing, or ameliorating a disease or condition in an individual, wherein TPO contributes to initiation of the disease or condition, the method comprising administering a compound comprising a safe TPO-specific inhibitor to the individual, thereby treating, preventing, or ameliorating the disease in the individual.

### Claim 2

A method of safely treating, preventing, or ameliorating a disease or condition in an individual, wherein TPO contributes to progression of the disease or condition, the method comprising administering a compound comprising a safe TPO-specific inhibitor to the individual, thereby treating, preventing, or ameliorating the disease in the individual.

### Claim 3

A method of safely treating, preventing, or ameliorating a disease or condition in an individual, wherein platelet activity contributes to initiation of the disease or condition, the method comprising administering a compound comprising a safe TPO-specific inhibitor to the individual, thereby treating, preventing, or ameliorating the disease in the individual.

### Claim 4

A method of safely treating, preventing, or ameliorating a disease or condition in an individual, wherein platelet activity contributes to progression of the disease or condition, the method comprising administering a compound comprising a safe TPO-specific inhibitor to the individual, thereby treating, preventing, or ameliorating the disease in the individual.

### Claim 5 (depends on 1, 2, 3, 4)

The method of any one of claims 1-4, wherein TPO activity adversely affects disease outcome in the individual.

### Claim 6 (depends on 1, 2, 3, 4, 5)

The method of any one of claims 1-5, wherein platelet activity adversely affects disease outcome in the individual.

### Claim 7 (depends on 1, 2, 3, 4, 5, 6)

The method of any one of claims 1-6, wherein the individual has a platelet count equal to or greater than about 100,000/μL, about 120,000/μL, about 150,000/μL, about 200,000/μL, about 250,000/μL, about 300,000/μL, about 350,000/μL, about 400,000/μL, about 450,000/μL, before administering.

### Claim 8 (depends on 1, 2, 3, 4, 5, 6, 7)

The method of any one of claims 1-7, wherein administering modulates TPO activity, platelet activity, or a combination thereof, in the individual.

### Claim 9 (depends on 1, 2, 3, 4, 5, 6, 7, 8)

The method of any one of claims 1-8, wherein the individual can benefit from platelet count reduction.

### Claim 10 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9)

The method of any one of claims 1-9, wherein administering reduces platelet count in the individual.

### Claim 11 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10)

The method of any one of claims 1-10, wherein administering reduces TOP activity, platelet activity, or a combination thereof, in the individual.

### Claim 12 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11)

The method of any one of claims 1-11, wherein the disease or condition is cancer, thrombocytosis, essential thrombocythemia, thrombosis, arterial thrombosis, venous thrombosis, inflammation, inflammatory disease, asthma, autoimmune disease, lupus, systemic lupus erythematosus, arthritis, psoriasis, diabetes, hyperglycemia, hyperinsulinemia, cardiovascular disease, or atherosclerosis.

### Claim 13 (depends on 12)

The method of claim 12, wherein the cancer is breast cancer, ovarian cancer, endometrial cancer, cervical cancer, prostate cancer, lung cancer, mesothelioma, gastrointestinal cancer, colon cancer, colorectal cancer, bladder cancer, liver cancer, esophageal cancer, pancreatic cancer, biliary tract cancer, sarcoma, chordoma, renal cancer, neuroblastoma, brain cancer, blood cancer, myeloma, B cell malignancy, lymphoma, or leukemia.

### Claim 14 (depends on 12, 13)

The method of claim 12 or 13, wherein administering the compound inhibits or reduces cancer cell proliferation, the number of primary tumors, individual tumor volume, total tumor volume, tumor growth, primary tumor intra-tumoral vessel density, primary tumor platelet deposition, and/or metastasis, or a combination thereof.

### Claim 15 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)

The method of any one of claims 1-14, wherein administering the safe TPO-specific inhibitor safely and partially reduces platelet count while leaving sufficient synthesis of TPO to maintain hemostatically safe platelet count.

### Claim 16 (depends on 15)

The method of claim 15, wherein synthesis of TPO is extrahepatic.

### Claim 17

A method of inhibiting expression or activity of TPO in a cell that expresses TPO receptor (MPL) comprising contacting the cell with a compound comprising a safe TPO-specific inhibitor, thereby inhibiting expression or activity of TPO in the cell.

### Claim 18 (depends on 17)

The method of claim 17, wherein the cell is a liver cell.

### Claim 19 (depends on 17)

The method of claim 17, wherein the cell is a cancer cell.

### Claim 20 (depends on 18, 19)

The method of claim 18 or 19, wherein the cell is in an individual.

### Claim 21 (depends on 20)

The method of claim 20, wherein the individual has, or is at risk of having, cancer.

### Claim 22 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21)

The method of any one of claims 1-21, wherein the individual is human.

### Claim 23 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)

The method of any one of claims 1-22, wherein the safe TPO-specific inhibitor selected from a nucleic acid, a polypeptide, an antibody, and a small molecule.

### Claim 24 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)

The method of any one of claims 1-23, wherein the compound comprises a modified oligonucleotide targeting TPO.

### Claim 25 (depends on 24)

The method of claim 24, wherein the compound is single-stranded.

### Claim 26 (depends on 24)

The method of claim 24, wherein the compound is double-stranded.

### Claim 27 (depends on 24, 25, 26)

The method of any one of claims 24-26, wherein the modified oligonucleotide is 12 to 30 linked nucleosides in length.

### Claim 28 (depends on 24, 25, 26, 27)

The method of any one of claims 24-27, wherein the modified oligonucleotide comprises at least one modified internucleoside linkage, at least one modified sugar moiety, or at least one modified nucleobase.

### Claim 29 (depends on 28)

The method of claim 28, wherein the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage, the at least one modified sugar is a bicyclic sugar or 2′-O-methyoxyethyl, and the at least one modified nucleobase is a 5-methylcytosine.

### Claim 30 (depends on 28, 29)

The method of claim 28 or 29, wherein at least one modified sugar comprises a 4′-CH(CH3)—O-2′ bridge or a 4′-(CH2)n—O-2′ bridge, wherein n is 1 or 2.

### Claim 31 (depends on 28, 29, 30)

The method of any one of claims 28-30, wherein each modified internucleoside is a phosphorothioate linkage.

### Claim 32 (depends on 28, 29, 30, 31)

The method of any one of claims 28-31, wherein each cytosine is a 5-methylcytosine.

### Claim 33 (depends on 24, 25, 26, 27, 28, 29, 30, 31, 32)

The method of any one of claims 24-32, wherein the modified oligonucleotide comprises:
a gap segment consisting of linked deoxynucleosides;
a 5′ wing segment consisting of linked nucleosides;
a 3′ wing segment consisting linked nucleosides;
wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

### Claim 34 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)

The method of any one of claims 1-33, wherein the compound is administered parenterally.

### Claim 35 (depends on 34)

The method of claim 34, wherein the compound is administered parenterally by subcutaneous or intravenous administration.

### Claim 36 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35)

The method of any one of claims 1-35, comprising co-administering the compound and at least one additional therapy.

### Claim 37 (depends on 36)

The method of claim 36, wherein the compound and the additional therapy are administered concomitantly.

### Claim 38 (depends on 37)

The method of claim 37, wherein the compound and the additional therapy are administered consecutively.

### Claim 39 (depends on 36, 37, 38)

The method of any one of claims 36-38, wherein the additional therapy is a secondary agent selected from a chemotherapeutic agent and a hormone therapy.

### Claim 40 (depends on 36, 37, 38)

The method of any one of claims 36-38, wherein the additional therapy is a secondary agent selected from an anti-HER2 antibody, a tyrosine kinase inhibitor, and a PARP inhibitor.

### Claim 41 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)

The method of any one of claims 1-40, wherein administering reduces an amount of a TPO RNA in a biological sample from the individual by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% relative to a reference amount in a reference sample.

### Claim 42 (depends on 41)

The method of claim 41, wherein the reference sample is a reference biological sample obtained from the individual before the administering.

### Claim 43 (depends on 41)

The method of claim 41, wherein the reference sample is a reference biological sample obtained from a control subject that does not have the disease or condition.

### Claim 44 (depends on 41, 42, 43)

The method of any one of claims 41-43, wherein the biological sample and the biological sample comprise blood, plasma, serum, urine, or a combination thereof.

### Claim 45

Use of a compound comprising a safe TPO-specific inhibitor for the manufacture or preparation of a medicament for treating a disease or condition that can benefit from platelet count reduction.

### Claim 46 (depends on 45)

The use of claim 45, wherein the disease or condition is thrombosis, arterial thrombosis, venous thrombosis, cancer, thrombocytosis, essential thrombocythemia, thrombosis, arterial thrombosis, venous thrombosis, inflammation, inflammatory disease, asthma, autoimmune disease, lupus, systemic lupus erythematosus, arthritis, psoriasis, diabetes, hyperglycemia, hyperinsulinemia, cardiovascular disease, or atherosclerosis.

### Claim 47

Use of a compound comprising a safe TPO-specific inhibitor for the treatment of a disease or condition that can benefit from platelet count reduction.

### Claim 48 (depends on 47)

The use of claim 47, wherein the disease or condition is cancer, thrombocytosis, essential thrombocythemia, thrombosis, arterial thrombosis, venous thrombosis, inflammation, inflammatory disease, asthma, autoimmune disease, lupus, systemic lupus erythematosus, arthritis, psoriasis, diabetes, hyperglycemia, hyperinsulinemia, cardiovascular disease, or atherosclerosis.

### Claim 49 (depends on 46, 48)

The use of claim 46 or 48, wherein the cancer is breast cancer, ovarian cancer, endometrial cancer, cervical cancer, prostate cancer, lung cancer, mesothelioma, gastrointestinal cancer, colon cancer, colorectal cancer, bladder cancer, liver cancer, esophageal cancer, pancreatic cancer, biliary tract cancer, sarcomas, chordoma, renal cancer, neuroblastoma, brain cancer, blood cancer, myeloma, multiple myeloma, a B cell malignancy, a lymphoma, a leukemia, or a myeloproliferative neoplasm.

### Claim 50 (depends on 45, 46, 47, 48, 49)

The use of any one of claims 45-49 wherein the safe TPO-specific inhibitor safely and partially reduces platelet count while leaving sufficient synthesis of TPO to maintain hemostatically safe platelet count.

### Claim 51 (depends on 50)

The use of claim 50, wherein the hemostatically safe platelet count is an extrahepatic platelet count.

### Claim 52 (depends on 45, 46, 47, 48, 49, 50, 51)

The use of any one of claims 45-51, wherein the compound is capable of reducing or inhibiting cancer cell proliferation, the number of primary tumors, individual tumor volume, total tumor volume, tumor growth, primary tumor intra-tumoral vessel density, primary tumor platelet deposition, or metastasis.

### Claim 53 (depends on 45, 46, 47, 48, 49, 50, 51, 52)

The use of any one of claims 45-52, wherein the safe TPO-specific inhibitor is selected from a nucleic acid, a polypeptide, an antibody, and a small molecule.

### Claim 54 (depends on 45, 46, 47, 48, 49, 50, 51, 52, 53)

The use of any of claims 45-53, wherein the compound comprises a modified oligonucleotide targeted to TPO.

### Claim 55 (depends on 54)

The use of claim 54, wherein the compound is single-stranded.

### Claim 56 (depends on 54)

The use of claim 54, wherein the compound is double-stranded.

### Claim 57 (depends on 54, 55, 56)

The use of any one of claims 54-56, wherein the modified oligonucleotide is 12 to 30 linked nucleosides in length.

### Claim 58 (depends on 54, 55, 56, 57)

The use of any one of claims 54-57, wherein the modified oligonucleotide comprises at least one modified internucleoside linkage, at least one modified sugar moiety, or at least one modified nucleobase.

### Claim 59 (depends on 58)

The use of claim 58, wherein the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage, the at least one modified sugar is a bicyclic sugar or 2′-O-methyoxyethyl, and the at least one modified nucleobase is a 5-methylcytosine.

### Claim 60 (depends on 58, 59)

The use of claim 58 or 59, wherein at least one modified sugar comprises a 4′-CH(CH3)—O-2′ bridge or a 4′-(CH2)n—O-2′ bridge, wherein n is 1 or 2.

### Claim 61 (depends on 58, 59, 60)

The use of any of claims 58-60, wherein each modified internucleoside is a phosphorothioate linkage.

### Claim 62 (depends on 58, 59, 60, 61)

The use of any of claims 58-61, wherein each cytosine is a 5-methylcytosine.

### Claim 63 (depends on 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62)

The use of any one of claims 41-62, wherein the modified oligonucleotide comprises:
a gap segment consisting of linked deoxynucleosides;
a 5′ wing segment consisting of linked nucleosides;
a 3′ wing segment consisting linked nucleosides;
wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.