# US 10,307,462 B2 · Compositions and methods for inducing thrombopoiesis

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- Number: US 10,307,462 B2
- Document ID: US10307462B2
- Status: Granted
- Kind: B2
- Application number: 14/534,394
- Filed: 2014-11-06
- Published: 2015-05-07
- Granted: 2019-06-04
- Earliest claimed: 2013-11-06
- Assignees: Hadasit Medical Research Services and Development Ltd.
- Applicants: Hadasit Medical Research Services and Development Ltd.
- Inventors: Dina Ben Yehuda; Ihab Abd-Elrahman; Riki Perlman; Marjorie Pick
- Web page: https://patentgrep.com/patent/US10307462B2

## CPC

- A61K 38/1761: {Apoptosis related proteins, e.g. Apoptotic protease-activating factor-1 (APAF-1), Bax, Bax-inhibitory protein(s)(BI; bax-I), Myeloid cell leukemia associated protein (MCL-1), Inhibitor of apoptosis [IAP] or Bcl-2}
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61K 35/545: Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- C12N 5/0644: {Platelets; Megakaryocytes}
- C12N 5/0647: {Haematopoietic stem cells; Uncommitted or multipotent progenitors}
- C12N 2501/48: Regulators of apoptosis
- C12N 2501/727: Kinases (EC 2.7.)
- C12N 2501/999: Small molecules not provided for elsewhere
- 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

Methods of inducing thrombopoiesis and/or treating thrombocytopenia in a subject are provided. Accordingly there is provided a method comprising contacting stem cells with a differentiation potential towards platelets or hematopoietic progenitor cells derived therefrom with Livin, thereby inducing thrombopoiesis. Also provided is a method comprising contacting cells with a differentiation potential towards platelets with tLivin, thereby inducing thrombopoiesis. Also provided are compositions and isolated population of cells for inducing thrombopoiesis and/or treating thrombocytopenia in a subject.

## Claims

### Claim 1

A method of inducing thrombopoiesis, the method comprising contacting stem cells with a differentiation potential towards platelets, said cells being selected from the group consisting of embryonic stem cells (ESCs), induced pluripotent stem cells (iPS), hematopoietic stem cells and hematopoietic progenitor cells derived therefrom with a platelet production stimulating factor selected from the group consisting of thrombopoietin (TPO), TPO agonist, stem cell factor (SCF) and Phorbol myristate acetate (PMA); and Livin in a formulation suitable for cell penetration, said Livin comprising:
(i) an anti-apoptotic Livin having an amino acid sequence as set forth in SEQ ID NO: 2 or SEQ ID NO: 6, wherein upon cleavage of said Livin by caspase 3 a pro-apoptotic tLivin is produced; or
(ii) a pro-apoptotic tLivin having an amino acid sequence as set forth in SEQ ID NO: 4 or SEQ ID NO: 8,
thereby inducing thrombopoiesis.

### Claim 2

A method of inducing thrombopoiesis, the method comprising contacting cells with a differentiation potential towards platelets with a platelet production stimulating factor selected from the group consisting of thrombopoietin (TPO), TPO agonist, stem cell factor (SCF) and Phorbol myristate acetate (PMA); and a pro-apoptotic tLivin having an amino acid sequence as set forth in SEQ ID NO: 4 or SEQ ID NO: 8 in a formulation suitable for cell penetration, thereby inducing thrombopoiesis.

### Claim 3 (depends on 2)

The method of claim 2, wherein said cells with differentiation potential towards platelets are selected from the group consisting of stem cells with a differentiation potential towards platelets, hematopoietic progenitor cells derived therefrom, and LAMA-84 cells.

### Claim 4 (depends on 1)

The method of claim 1, wherein said hematopoietic progenitor cells comprise megakaryocytes.

### Claim 5 (depends on 1)

The method of claim 1, wherein said stem cells with a differentiation potential towards platelets or hematopoietic progenitor cells are comprised in a biological sample.

### Claim 6 (depends on 5)

The method of claim 5, wherein said biological sample is selected from the group consisting of cord blood, peripheral blood (PB), peripheral blood mononuclear cells (PBMCs) and bone marrow.

### Claim 7 (depends on 5)

The method of claim 5, wherein said biological sample comprises PBMCs.

### Claim 8 (depends on 1)

The method of claim 1, wherein said Livin is said tLivin.

### Claim 9 (depends on 1)

The method of claim 1, wherein said formulation is selected from the group consisting of a liposome, a nanoparticle or a cell penetrating peptide.

### Claim 10 (depends on 1)

The method of claim 1, wherein said stem cells are embryonic stem cells (ESC) or induced pluripotent stem cells (iPS).

### Claim 11 (depends on 1)

The method of claim 1, wherein said cells are selected from the group consisting of embryonic stem cells (ESCs), hematopoietic stem cells and hematopoietic progenitor cells derived therefrom.

### Claim 12 (depends on 1)

The method of claim 1, wherein said cells are hematopoietic CD34+ stem cells.

### Claim 13 (depends on 1)

The method of claim 1, wherein said cells are hematopoietic progenitor cells selected from the group consisting of myeloid precursor cells and megakaryocytes.