# US 10,245,286 B2 · Methods and compositions for modulating angiogenesis and vasculogenesis

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- Number: US 10,245,286 B2
- Document ID: US10245286B2
- Status: Granted
- Kind: B2
- Application number: 15/608,656
- Filed: 2017-05-30
- Published: 2017-09-14
- Granted: 2019-04-02
- Earliest claimed: 2012-01-27
- Assignees: SanBio, Inc.
- Applicants: SanBio, Inc.
- Inventors: Monique Dao; Ciara Tate; Casey C. Case
- Web page: https://patentgrep.com/patent/US10245286B2

## CPC

- A61K 35/34: Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes (vascular smooth muscle A61K35/44)
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61K 35/44: Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
- A61K 38/1866: {Vascular endothelial growth factor [VEGF]}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61L 27/3834: {Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells}
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 25/00: Drugs for disorders of the nervous system
- C12N 5/0662: {Stem cells}
- C12N 5/0663: {Bone marrow mesenchymal stem cells (BM-MSC)}
- C12N 5/0668: {Mesenchymal stem cells from other natural sources}
- C12N 5/0697: {Artificial constructs associating cells of different lineages, e.g. tissue equivalents (blood vessels C12N5/0691)}
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- C12N 2501/42: Notch; Delta; Jagged; Serrate
- C12N 2502/1347: Smooth muscle cells
- C12N 2502/1352: Mesenchymal stem cells
- C12N 2502/28: Vascular endothelial cells
- C12N 2510/02: Cells for production
- C12N 2533/90: Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
- 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)
- A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K)
- 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

Disclosed herein are methods and compositions for stimulating angiogenesis, using cells descended from marrow adherent stromal cells that have been transfected with sequences encoding a Notch intracellular domain. Applications of these methods and compositions include treatment of ischemic disorders such as stroke.

## Claims

### Claim 1

A method for treating an ischemic disorder in a subject, wherein the ischemic disorder occurs outside the central nervous system, the method comprising:
administering, to a site of ischemic disorder in the subject that is outside the central nervous system, a therapeutically effective amount of cells, wherein the cells are made by a method comprising:
(a) providing a culture of mesenchymal stem cells,
(b) contacting the cell culture of step (a) with a polynucleotide comprising sequences encoding a Notch intracellular domain (NICD) wherein said polynucleotide does not encode a full-length Notch protein,
(c) selecting cells that comprise the polynucleotide of step (b), and
(d) further culturing the selected cells of step (c) in the absence of selection and in the absence of trophic factors;
wherein the cells produce angiogenin, angiopoietin-2 (ANG-2) and heparin-binding epidermal growth factor-like growth factor (HB-EGF) at higher levels than do the mesenchymal stem cells of step (a).

### Claim 2 (depends on 1)

The method of claim 1, wherein the ischemic disorder is selected from the group consisting of cardiac ischemia, ischemia of the bowel, ischemia of the limb, cutaneous ischemia, retinal ischemia and cerebral palsy.

### Claim 3 (depends on 2)

The method of claim 2, wherein the ischemia of the bowel is ischemic colitis or mesenteric ischemia.

### Claim 4 (depends on 1)

The method of claim 1, wherein the cells produce one or more additional angiogenic factors selected from the group consisting of basic fibroblast growth factor, leptin, platelet-derived growth factor-BB, hepatocyte growth factor, vascular endothelial growth factor and placental growth factor.

### Claim 5 (depends on 1)

The method of claim 1, wherein administration results in augmentation of blood vessel sprouting.

### Claim 6 (depends on 1)

The method of claim 1, wherein administration results in prevention of endothelial cell death.

### Claim 7 (depends on 1)

The method of claim 1, wherein administration results in enhancement of endothelial cell survival.

### Claim 8 (depends on 1)

The method of claim 1, wherein administration results in stimulation of endothelial cell proliferation.

### Claim 9 (depends on 1)

The method of claim 1, wherein administration results in enhancement of vessel branching.

### Claim 10

A combination for use in treating an ischemic disorder in a subject, wherein the ischemic disorder occurs outside the central nervous system, wherein the combination comprises:
(a) a first pro-angiogenic agent comprising cells made by a method comprising:
(i) providing a culture of mesenchymal stem cells,
(ii) contacting the cell culture of step (i) with a polynucleotide comprising sequences encoding a Notch intracellular domain (NICD) wherein said polynucleotide does not encode a full-length Notch protein,
(iii) selecting cells that comprise the polynucleotide of step (ii), and
(iv) further culturing the selected cells of step (iii) in the absence of selection and in the absence of trophic factors;
wherein the cells produce angiogenin, angiopoietin-2 (ANG-2) and heparin-binding epidermal growth factor-like growth factor (HB-EGF) at higher levels than do the mesenchymal stem cells of step (i); and
(b) a second pro-angiogenic agent.

### Claim 11 (depends on 10)

The combination of claim 10, wherein the second pro-angiogenic agent is a polypeptide.

### Claim 12 (depends on 11)

The combination of claim 11, wherein the polypeptide is a transcription factor that activates transcription of a pro-angiogenic protein.

### Claim 13 (depends on 12)

The combination of claim 12, wherein the pro-angiogenic protein is vascular endothelial growth factor (VEGF).

### Claim 14 (depends on 13)

The combination of claim 13, wherein the transcription factor is a non-naturally-occurring zinc finger protein that activates transcription of the VEGF gene.

### Claim 15 (depends on 10)

The combination of claim 10, wherein the second pro-angiogenic agent is a nucleic acid.

### Claim 16 (depends on 15)

The combination of claim 15, wherein the nucleic acid encodes a transcription factor that activates transcription of a pro-angiogenic protein.

### Claim 17 (depends on 16)

The combination of claim 16, wherein the pro-angiogenic protein is vascular endothelial growth factor (VEGF).

### Claim 18 (depends on 17)

The combination of claim 17, wherein the transcription factor is a non-naturally-occurring zinc finger protein that activates transcription of the VEGF gene.

### Claim 19 (depends on 10)

A method for use in treating an ischemic disorder in a subject, wherein the ischemic disorder occurs outside the central nervous system, the method comprising administering to the subject a therapeutically effective amount of the combination of claim 10.

### Claim 20 (depends on 19)

The method of claim 19, wherein administration results in augmentation of angiogenesis.

### Claim 21 (depends on 20)

The method of claim 20, wherein said augmentation of angiogenesis comprises a process selected from the group consisting of one or more of prevention of endothelial cell death, enhancement of endothelial cell survival, stimulation of endothelial cell proliferation, enhancement of blood vessel sprouting and enhancement of blood vessel branching.