# US 2014/0193376 A1 · Combined chemical modification of sphingosine-1-phosphate (s1p) and cxcr4 signalling pathways for hematopoietic stem cell (hsc) mobilization and engraftment

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- Number: US 2014/0193376 A1
- Document ID: US20140193376A1
- Status: Published application
- Kind: A1
- Application number: 14/126,768
- Filed: 2012-06-18
- Published: 2014-07-10
- Earliest claimed: 2011-06-16
- Assignees: Children's Medical Center Corporation
- Applicants: Children's Medical Center Corporation
- Inventors: Leonard I Zon; Owen J. Tamplin
- Web page: https://patentgrep.com/patent/US20140193376A1

## CPC

- A61K 35/14: Blood; Artificial blood (perfluorocarbons A61K31/02; umbilical cord blood A61K35/51; haemoglobin A61K38/42)
- A61K 31/137: Arylalkylamines, e.g. amphetamine, epinephrine, salbutamol, ephedrine {or methadone}
- A61K 31/138: Aryloxyalkylamines, e.g. propranolol, tamoxifen, phenoxybenzamine (atenolol A61K31/165; pindolol A61K31/404; timolol A61K31/5377)
- A61K 31/381: having five-membered rings
- A61K 31/395: having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K 31/4245: Oxadiazoles
- A61K 31/662: Phosphorus acids or esters thereof having P—C bonds, e.g. foscarnet, trichlorfon
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61K 38/04: Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof ({enzyme inhibitors A61K38/005;} gastrins {A61K38/2207}, somatostatins A61K38/31, melanotropins A61K38/34 {; protease inhibitors A61K38/55})
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- 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

The present embodiments provide for combinations of modulators that increase hematopoietic stem cell engraftment or increase mobilization in vivo. Methods and compositions for modulating the mobilization of stem cells, particularly for promoting or increasing the mobilization of hematopoietic stem cells (HSCs) from the bone marrow to the peripheral blood are disclosed. One aspect of the invention relates to the use of a CXCR4 antagonist that act in concert with specific molar ratios of S1P receptor 1 (S1PR1) modulator agents to promote HSC mobilization. The invention also relates to methods of using these combinations of CXCR4 antagonists and S1PR1 modulator agents for enhancing the mobilization of hematopoietic stem cells when harvesting of the stem cells, for example for the treatment of diseases, disabilities or conditions whereby transplantation of such cells would be beneficial in ameliorating a symptom associated with such diseases, disabilities or conditions. Another aspect of the invention relates to the use of a CXCR4 antagonist that act in concert with different, but specific molar ratios of S1P receptor 1 (S1PR1) modulator agents to promote HSC engraftment and methods for promoting HSC engraftment in a subject in need thereof, e.g., a recipient subject of a bone marrow or HSC transplant. Methods of screening for novel agents and pharmaceutical compositions comprising these agents are also disclosed.

## Claims

### Claim at position 1

-84. (canceled)

### Claim 85

A method comprising administering to a subject in need of hematopoietic stem cell (HSC) engraftment a CXCR4 antagonist and an agent selected from the group consisting of:
a) (R)-3-Amino-4-(3-hexylphenylamino)-4-oxobutylphosphonic acid (W146), wherein the ratio of the concentration of the CXCR4 antagonist to the concentration of the W146 compound is (0.5-0.6):1.0;
b) 3-[[2-[4-phenyl-3-(trifluoromethyl)phenyl]-1-benzothiophen-5-yl]methylamino]propanoic acid (AUY954), wherein the ratio of the concentration of the CXCR4 antagonist to the concentration of the AUY954 compound is (0.4-2.5):1.0;
c) 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol (FTY720), wherein the ratio of the concentration of the CXCR4 antagonist to the concentration of the 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol compound is (0.4-3.0):1.0; and
d) 5-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-3-[3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole (SEW2871), wherein the ratio of the concentration of the CXCR4 antagonist to the concentration of the 5-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-3-[3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole compound is (0.25-1.0):1.0.

### Claim 86 (depends on 85)

The method of claim 85, wherein the CXCR4 antagonist is 1,1′-[1,4-Phenylenebis(methylene)]bis[1,4,8,11-tetraazacyclotetradecane] (AMD3100) or T-140.

### Claim 87 (depends on 86)

The method of claim 86, wherein the AMD3100 is at a concentration of between 2-10 μM and the concentration of W146 is between of 4-20 μM.

### Claim 88 (depends on 85)

The method of claim 85, wherein the subject is administered a population of HSCs prior to, during or after administration of the CXCR4 antagonist and W146, AUY954, FTY720 or SEW287Y.

### Claim 89 (depends on 86)

The method of claim 86, wherein the AMD3100 is at a concentration of between 5-20 μM and the concentration of AUY954 is between of 5-25 μM.

### Claim 90 (depends on 86)

The method of claim 86, wherein the concentration of AMD3100 is between 10-25 μM and the concentration of FTY720 is between of 2-10 μM.

### Claim 91 (depends on 86)

The method of claim 86, wherein the AMD3100 or SEW2871 is at a concentration of between 5-20 μM.

### Claim 92 (depends on 85)

The method of claim 85, wherein the subject is a candidate for bone marrow or stem cell transplantation, or a subject that has received bone marrow ablating chemotherapy or irradiation therapy.

### Claim 93

A method comprising administering to a subject in need of hematopoietic stem cell (HSC) mobilization a CXCR4 antagonist and an agent selected from the group consisting of:
a) (R)-3-Amino-4-(3-hexylphenylamino)-4-oxobutylphosphonic acid (W146), wherein the ratio of the concentration of the CXCR4 antagonist to the concentration of the W146 compound is (1.25-2.0):1;
b) 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol (FTY720), wherein the ratio of the concentration of the CXCR4 antagonist to the concentration of the 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol compound is 5:1; and
c) 5-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-3-[3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole (SEW2871), wherein the ratio of the concentration of the CXCR4 antagonist to the concentration of the 5-[4-phenyl-5-(trifluoromethyl)-2-thienyl]-3-[3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole compound is (1.25-5.0):1.0.

### Claim 94 (depends on 93)

The method of claim 93, wherein the CXCR4 antagonist is 1,1′-[1,4-Phenylenebis(methylene)]bis[1,4,8,11-tetraazacyclotetradecane] (AMD3100) or T-140.

### Claim 95 (depends on 94)

The method of claim 94, wherein the AMD3100 is at a concentration of between 5-20 μM and the concentration of W146 is between of 2-10 μM.

### Claim 96 (depends on 93)

The method of claim 93, wherein a population of HSCs are obtained from the subject during or after administration of the CXCR4 antagonist and W146, FTY720, or SEW2871.

### Claim 97 (depends on 96)

The method of claim 96, wherein the population of HSCs is obtained from peripheral blood, cord blood, bone marrow, amniotic fluid, or placental blood.

### Claim 98 (depends on 94)

The method of claim 94, wherein the AMD3100 is at a concentration of between 10-50 μM and the concentration of FTY720 is between of 2-10 μM.

### Claim 99 (depends on 94)

The method of claim 94, wherein the AMD3100 is at a concentration of between 10-50 μM and the concentration of SEW2871 is between 6-30 μM.

### Claim 100 (depends on 94)

The method of claim 94, wherein the AMD3100 is at a concentration of between 2 mg/kg and 10 mg/kg, SEW2871 is between 1.2 mg/kg and 6 mg/kg.

### Claim 101 (depends on 94)

The method of claim 94, wherein the population of HSCs obtained from the subject are c cryopreserved.

### Claim 102 (depends on 93)

The method of claim 93, wherein the population of HSCs obtained from the subject are administered to a subject in a bone marrow or stem cell transplantation.

### Claim 103 (depends on 93)

The method of claim 93, wherein the subject is a bone marrow donor.

### Claim 104

A pharmaceutical composition or kit comprising at least one CXCR4 antagonist and an agent selected from the group consisting of:
a) W146 in an effective amount to promote HSC engraftment or migration in a subject;
b) AUY in an effective amount to promote HSC engraftment in a subject;
c) FTY720 in an effective amount to promote HSC engraftment or migration in a subject; and
d) SEW2781 in an effective amount to promote HSC engraftment or migration in a subject.