# US 11,992,506 B2 · Stem cell derived pancreatic islet differentiation

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

- Number: US 11,992,506 B2
- Document ID: US11992506B2
- Status: Granted
- Kind: B2
- Application number: 18/051,721
- Filed: 2022-11-01
- Published: 2023-07-13
- Granted: 2024-05-28
- Earliest claimed: 2021-11-01
- Assignees: Vertex Pharmaceuticals Incorporated
- Applicants: Vertex Pharmaceuticals Incorporated
- Inventors: George Harb; Chunhui Xie; Bryce W. Carey; Aleksander Szymaniak; Christopher Thanos; Evrett Thompson; Rebecca Chinn; Suyash Raj
- Web page: https://patentgrep.com/patent/US11992506B2

## CPC

- A61K 35/39: Pancreas; Islets of Langerhans (Langerhans cells of epidermis A61K35/36)
- A61K 31/4704: 2-Quinolinones, e.g. carbostyril
- A61K 38/28: Insulins
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- C12N 5/0676: {Pancreatic cells}
- C12N 5/0678: {Stem cells; Progenitor cells; Precursor cells}
- C12N 2500/50: Soluble polymers, e.g. polyethyleneglycol [PEG]
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- C12N 2501/117: Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
- C12N 2501/16: Activin; Inhibin; Mullerian inhibiting substance
- C12N 2501/42: Notch; Delta; Jagged; Serrate
- C12N 2506/45: from artificially induced pluripotent stem cells
- C12N 2533/30: Synthetic polymers
- 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

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic islet cells. In some aspects, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some aspects, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

## Claims

### Claim 1

An in vitro composition comprising a plurality of non-native cells prepared in vitro; wherein:
a) at least 30% of the cells in the composition are NKX6.1-positive, ISL1-positive cells;
b) at least 25% of the cells in the composition are NKX6.1-negative, ISL1-positive cells;
c) there are more NKX6.1-positive, ISL1-positive cells than NKX6.1-negative, ISL1-positive cells in the composition; and
d)
i) less than 6% of the cells in the composition are NKX6.1-negative, ISL1-negative cells; and/or
ii) between 9-25% of the cells in the composition are NKX6.1-positive, ISL1-negative cells.

### Claim 2 (depends on 1)

The composition of claim 1, wherein 30-60% of the cells in the plurality of cells are NKX6.1-positive, ISL1-positive cells, and wherein 20-50% of the cells in the plurality of cells are NKX6.1-negative, ISL1-positive cells.

### Claim 3 (depends on 1)

The composition of claim 1, wherein 35-55% of the cells in the plurality of cells are NKX6.1-positive, ISL1-positive cells, and wherein 25-45% of the cells in the plurality of cells are NKX6.1-negative, ISL1-positive cells.

### Claim 4 (depends on 1)

The composition of claim 1, wherein 1-6% of the cells in the plurality of cells are NKX6.1-negative, ISL1-negative cells.

### Claim 5 (depends on 1)

The composition of claim 1, wherein between 2-25% of the cells in the plurality of cells are NKX6.1-positive, ISL1-negative cells, and wherein 1-6% of the cells in the plurality of cells are NKX6.1-negative, ISL1-negative cells.

### Claim 6 (depends on 1)

The composition of claim 1, wherein between 9-25% of the cells in the plurality of cells are NKX6.1-positive, ISL1-negative cells.

### Claim 7 (depends on 1)

The composition of claim 1, wherein 35-55% of the cells in the plurality of cells are NKX6.1-positive, ISL1-positive cells, wherein 25-45% of the cells in the plurality of cells are NKX6.1-negative, ISL1-positive cells; and wherein 1-6% of the cells in the plurality of cells are NKX6.1-negative, ISL1-negative cells.

### Claim 8 (depends on 1)

The composition of claim 1, wherein the plurality of cells are in a cell cluster.

### Claim 9 (depends on 1)

The composition of claim 1, wherein the plurality of cells are in a plurality of cell clusters.

### Claim 10 (depends on 8)

The composition of claim 8, wherein the cell cluster comprises between 500-2500 cells.

### Claim 11 (depends on 9)

The composition of claim 9, wherein the plurality of cell clusters each comprise between 500-2500 cells.

### Claim 12 (depends on 8)

The composition of claim 8, wherein the cluster comprises between 500-2500 NKX6.1-positive, ISL1-positive cells.

### Claim 13 (depends on 9)

The composition of claim 9, wherein the plurality of cell clusters each comprise between 500-2500 NKX6.1-positive, ISL1-positive cells.

### Claim 14 (depends on 8)

The composition of claim 8, wherein the cell cluster has a diameter between 80-270 microns.

### Claim 15 (depends on 9)

The composition of claim 9, wherein the plurality of cell clusters each have a diameter between 80-270 microns.

### Claim 16 (depends on 1)

The composition of claim 1, wherein the composition comprises cells that have been genetically modified to reduce expression of any one or more of the following genes: B2M, CIITA, CXCL10, renalase, HLA-A, HLA-B, HLA-C, NLRC5, ABO, RHD, FUT1, KDMSD, PDGFRa, OLIG2, and/or GFAP as compared to a cell that has not been genetically modified.

### Claim 17 (depends on 1)

The composition of claim 1, wherein the composition comprises cells that have been genetically modified to increase expression of any or more of the following genes: CD47, PDL1, PDL2, HLA-E, HLA-G, CD46, CD55, CD59 and CTLA, relative to cells that have not been genetically modified.

### Claim 18 (depends on 1)

The composition of claim 1, wherein the composition further comprises a medium that comprises a sugar at a concentration of between about 0.05% and about 1.5%.

### Claim 19 (depends on 1)

The composition of claim 1, wherein the composition further comprises a CMRL medium or a HypoThermosol® FRS Preservation Media.

### Claim 20 (depends on 1)

The composition of claim 1, wherein the composition is in a device, wherein the device comprises a semipermeable membrane that is configured to retain the cells in the device and permit passage of insulin secreted by the cells out of the device.

### Claim 21 (depends on 20)

The composition of claim 20, wherein the semipermeable membrane can be made of poly(lactide) (PLA), poly(glycolic acid) (PGA), poly(lactide-co-glycolide) (PLGA), and other polyhydroxyacids, poly(caprolactone), polycarbonates, polyamides, polyanhydrides, polyphosphazene, polyamino acids, polyortho esters, polyacetals, polycyanoacrylates, biodegradable polyurethanes, albumin, collagen, fibrin, polyamino acids, prolamines, alginate, agarose, agarose with gelatin, dextran, polyacrylates, ethylene-vinyl acetate polymers and other acyl-substituted cellulose acetates and derivatives thereof, polyurethanes, polystyrenes, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonated polyolefins, polyethylene oxide, or any combinations thereof.

### Claim 22 (depends on 1)

The composition of claim 1, wherein at least 65-85% of the cells express ISL1.

### Claim 23 (depends on 1)

The composition of claim 1, wherein the plurality of cells comprise 0.1-5% ghrelin-positive cells.

### Claim 24 (depends on 1)

The composition of claim 1, wherein the plurality of cells comprise 2-10% somatostatin-positive cells.

### Claim 25 (depends on 1)

The composition of claim 1, wherein the composition comprises NKX6.1-positive, ISL1-positive cells that display a glucose stimulated insulin secretion (GSIS) response similar or superior to that of an endogenous mature β cell if tested in vitro.

### Claim 26 (depends on 1)

The composition of claim 1, wherein the composition comprises NKX6.1-positive, ISL1-positive cells that also express PC2, MNX1, or ABCC8.

### Claim 27 (depends on 1)

The composition of claim 1, wherein the C-peptide content per 1,000 of the in vitro differentiated cells is at least 300 pM, optionally wherein the C-peptide content per 1,000 of the in vitro differentiated cells is at least 400 pM.

### Claim 28 (depends on 1)

A method of treating a subject having Type 1 Diabetes comprising administering to the subject the composition of claim 1.

### Claim 29 (depends on 28)

The method of claim 28, wherein the composition is administered in a device, wherein the device comprises a semipermeable membrane that is configured to retain the cells in the device and permit passage of insulin secreted by the cells out of the device.

### Claim 30 (depends on 28)

The method of claim 28, wherein the composition is administered via the portal vein.