# US 12,215,355 B2 · Stem cell derived islet differentiation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,215,355 B2 - Document ID: US12215355B2 - Status: Granted - Kind: B2 - Application number: 17/171,497 - Filed: 2021-02-09 - Published: 2021-08-05 - Granted: 2025-02-04 - Earliest claimed: 2018-08-10 - Assignees: Vertex Pharmaceuticals Incorporated - Applicants: Vertex Pharmaceuticals Incorporated - Inventors: Felicia J. Pagliuca; George Harb; Lillian Ye - Web page: https://patentgrep.com/patent/US12215355B2 ## CPC - C12N 5/0676: {Pancreatic cells} - C12N 5/0678: {Stem cells; Progenitor cells; Precursor cells} - A61K 35/39: Pancreas; Islets of Langerhans (Langerhans cells of epidermis A61K35/36) - A61P 5/50: for increasing or potentiating the activity of insulin - C07K 14/4702: {Regulators; Modulating activity} - A61K 2035/126: {Immunoprotecting barriers, e.g. jackets, diffusion chambers} - C12N 2500/46: Amines, e.g. putrescine - C12N 2501/11: Epidermal growth factor [EGF] - C12N 2501/117: Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12) - C12N 2501/119: Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10 - C12N 2501/15: Transforming growth factor beta (TGF-β) - C12N 2501/155: Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor - C12N 2501/16: Activin; Inhibin; Mullerian inhibiting substance - C12N 2501/19: Growth and differentiation factors [GDF] - C12N 2501/415: Wnt; Frizzeled - C12N 2501/727: Kinases (EC 2.7.) - C12N 2501/999: Small molecules not provided for elsewhere - C12N 2506/45: from artificially induced pluripotent stem cells - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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 - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract Provided herein are methods of producing β cells and precursors thereof utilizing a Wnt signaling inhibitor or PKC activator, or both. Also provided herein are in vitro cultures comprising said cells, methods of treating a subject with a disease characterized by high blood sugar levels over a prolonged period of time by administering said cells, and devices for encapsulating said cells. ## Claims ### Claim 1 A composition comprising PDX1-positive, NKX6.1-positive pancreatic progenitor cells, a WNT signaling inhibitor, and one or more of a thyroid receptor signaling pathway activator, a protein kinase C activator, or a transforming growth factor (TGF)-β signaling pathway inhibitor, wherein the WNT signaling inhibitor is present at a concentration of 0.1 μM to 10 μM. ### Claim 2 (depends on 1) The composition of claim 1, wherein said WNT signaling inhibitor comprises a tankyrase inhibitor that inhibits expression or activity of at least one tankyrase (TNKS) protein. ### Claim 3 (depends on 2) The composition of claim 2, wherein said tankyrase signaling inhibitor is selected from the group consisting of: AZ 6102, JW55, 11N64, IWR-1-endo, TC-E5001, WIKI4, TNKS 22, TNKS 49, 2X-121 (E7449), XAV-939, G007-LK, NVP-TNKS656, decernotinib (VX-509), vismodegib (GDC-0449), IM-12, GSK429286A, INO-1001, Ofloxacin, TG101209, FG-4592, I-BET-762, LY2157299, MK-0752, Wnt-C59 (C59), MC1568, Pacritinib (SB1518), SB415286, Drocinostat, IWR-1-endo, Norfloxacin, SH-4-54, Nexturastat A, SB216763, UNC0.79, dephnetin, GF109203X, RepSox, Sotrastaurin, SB431542, tofacitinib (CP-690550), AG-14361, CI994 (tacedinaline), Ro 31-8220 mesylate, resveratrol, NVP-TNKS656, and YO-01027. ### Claim 4 (depends on 1) The composition of claim 1, wherein said WNT signaling inhibitor comprises NVP-TNKS656. ### Claim 5 (depends on 1) The composition of claim 1, wherein said WNT signaling inhibitor comprises XAV-939. ### Claim 6 (depends on 1) The composition of claim 1, wherein said composition comprises about 1 μM to about 5 μM of said WNT signaling inhibitor. ### Claim 7 (depends on 1) The composition of claim 1, wherein said composition comprises about 0.5 μM of said WNT signaling inhibitor. ### Claim 8 (depends on 1) The composition of claim 1, further comprising PDX1-positive, NKX6.1-positive, ISL1-positive cells. ### Claim 9 (depends on 1) The composition of claim 1, wherein said composition further comprises: a sonic hedgehog pathway inhibitor selected from the group consisting of: SANT1, SANT2, SANT4, Cur61414, forskolin, tomatidine, AY9944, triparanol, and cyclopamine. ### Claim 10 (depends on 1) The composition of claim 1, wherein said composition further comprises a retinoic acid signaling pathway activator selected from the group consisting of: retinoic acid, CD1530, AM580, TTHRB, CD437, Ch55, BMS961, AC261066, AC55649, AM80, BMS753, tazarotene, adapalene, and CD2314. ### Claim 11 (depends on 1) The composition of claim 1, wherein said composition further comprises a γ-secretase inhibitor comprising XXI or DAPT. ### Claim 12 (depends on 1) The composition of claim 1, wherein said composition further comprises a growth factor from the epidermal growth factor family comprising betacellulin or EGF. ### Claim 13 (depends on 1) The composition of claim 1, wherein said composition further comprises a bone morphogenetic protein signaling pathway inhibitor comprising LDN193189. ### Claim 14 (depends on 1) The composition of claim 1, wherein said composition comprises said TGF-β signaling pathway inhibitor, and wherein said TGF-β signaling pathway inhibitor is selected from the group consisting of: Alk5i II, A83-01, SB431542, D4476, GW788388, LY364947, LY580276, SB505124, GW6604, SB-525334, and SD-208. ### Claim 15 (depends on 1) The composition of claim 1, wherein said composition comprises said thyroid receptor signaling pathway activator, and wherein said thyroid receptor signaling pathway activator comprises T3 or GC-1. ### Claim 16 (depends on 1) The composition of claim 1, wherein said composition further comprises a protein kinase inhibitor selected from the group consisting of: staurosporine, Ro-31-8220, a bisindolylmaleimide (Bis) compound, 10′-{5″-[(methoxycarbonyl) amino]-2″-methyl}-phenylaminocarbonylstaurosporine, a staralog, and cgp41251. ### Claim 17 (depends on 1) The composition of claim 1, wherein said composition further comprises a ROCK inhibitor selected from the group consisting of Thiazovivin, Y-27632, and Fasudil/HA1077. ### Claim 18 (depends on 1) The composition of claim 1, wherein said composition further comprises SANT1. ### Claim 19 (depends on 1) The composition of claim 1, wherein said composition further comprises Alk5i II. ### Claim 20 (depends on 1) The composition of claim 1, wherein said composition further comprises staurosporine.