Compositions and methods for providing chemical protective effects
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- G TECH BIO LLC
- Filing Date
- 2026-02-06
- Publication Date
- 2026-06-09
Smart Images
Figure 2026094171000001_ABST
Abstract
Claims
1. A method for providing or improving chemoprotection in target cells, comprising providing the target cells with a heterologous glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function.
2. A method for providing or improving chemical protection in a patient in need thereof, comprising administering to the patient cells expressing a heterologous glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides the equivalent function.
3. The method according to claim 1 or 2, wherein post-transplant in vivo selection is possible by transgenetic expression of the glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function.
4. A method for conferring chemotherapy-resistant cells to a patient, the method comprising administering to the patient cells expressing heterologous glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function.
5. The method according to claim 4, wherein the cells express heterologous GCLM.
6. A method for treating a patient having cancer, the method comprising administering to the patient, together with a non-myeloablative or myeloablative dose of a chemotherapeutic agent, a pharmaceutically acceptable composition comprising cells expressing heterologous glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function.
7. The method according to claim 6, wherein the cells heterologously express GCLM.
8. The method according to claim 6 or 7, further comprising administering to the patient cells expressing a heterologous glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides the equivalent function, before, after, or concurrently with the chemotherapeutic agent.
9. The method according to any one of claims 4 to 8, further comprising isolating stem cells or immune cells from the patient.
10. The method according to claim 9, wherein the isolated stem cells or immune cells are modified to express the heterologous glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides the equivalent function.
11. The isolated stem cells or immune cells are modified to express heterologous GCLM. The method according to claim 10.
12. The method according to claim 10 or 11, wherein the cells are modified by contacting the isolated cells with an expression vector comprising GCLM or glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides the equivalent function.
13. The method according to claim 12, wherein the vector is a viral vector or a nonviral vector.
14. The method according to claim 12, wherein the vector is a lentiviral vector, an adenovirus vector, a plasmid, a retrovirus, a transposon, an episome expression vector, a modified RNA, or any combination thereof.
15. The method according to any one of claims 1 to 14, wherein the cells are stem cells or immune cells.
16. The method according to claim 15, wherein the stem cells are fetal stem cells, umbilical cord blood-derived stem cells, hematopoietic stem cells (HSCs), pluripotent stem cells (PSCs), induced PSCs (iPSCs), embryonic stem cells (ESCs), or cells derived therefrom, such as CD34+ cells, CD90+ cells, CD45+ cells, CD17+ cells, CD45RA- cells, or any combination thereof.
17. The method according to claim 15, wherein the immune cell is a T cell.
18. The method according to any one of claims 1 to 16, wherein the modified cells are autologous to the patient, homogeneous to the patient, or a combination thereof.
19. The method according to any one of claims 1 to 18, wherein the patient is receiving gene therapy or CAR-T cell therapy.
20. The method according to any one of claims 1 to 19, wherein the chemical protection provided is against busulfan and / or naphthalene.
21. The method according to any one of claims 1 to 20, further comprising administering to the patient a myeloablative or non-myeloablative dose of busulfan and / or naphthalene.
22. A method for performing a bone marrow transplant on a patient, The method comprising administering to the patient a population of busulfan-resistant modified cells and at least one myeloablative dose of busulfan.
23. The method according to claim 22, wherein the population of busulfan-resistant modified cells comprises heterogenes encoding glutamate cysteine ligase (GCL) modifier subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function.
24. The method according to claim 22, wherein the heterogene encodes GCLM.
25. The method according to claim 22, wherein the population of busulfan-resistant modified cells expresses GCLM.
26. The method according to claim 22, wherein the busulfan resistance of the busulfan-resistant modified cells is conferred by the expression of GCLM.
27. The method according to claim 22, wherein more than 50% of the bone marrow of the patient is replaced with the busulfan-resistant modified cells or cells derived therefrom within six months.
28. The method according to claim 22, wherein the patient has HIV, cancer, A1AT deficiency, WAS, Flurer syndrome, Hunter syndrome, Pompe disease, Fabry disease, mucopolysaccharidosis, MPS 1 H / S (Flurer / Chaillet syndrome), MPS I H (Flurer disease), MPS II- (Hunter syndrome), MPS III A, B, C, and D (Sanfilippo syndrome), MPS I S (Chaillet syndrome), MPS IV A and B (Morcchio syndrome), MPS IX (hyaluronidase deficiency), MPS VII (Sly syndrome), MPS VI (Maroto-Lamy syndrome), lysosome storage disorder, and childhood cerebral adrenoleukodystrophy (cALD).
29. The method according to claim 22, wherein the busulfan resistance of the modified cells is transient.
30. The method according to claim 22, wherein the cells are stem cells or immune cells.
31. The method according to claim 30, wherein the stem cells are fetal stem cells, umbilical cord blood-derived stem cells, hematopoietic stem cells (HSCs), pluripotent stem cells (PSCs), induced PSCs (iPSCs), embryonic stem cells (ESCs), or cells derived therefrom, such as CD34+ cells, CD90+ cells, CD45+ cells, CD17+ cells, CD45RA- cells, CD38-, or any combination thereof.
32. The method according to claim 30, wherein the immune cell is a T cell.
33. The method according to claim 22, wherein the modified cells are autologous to the patient, homogeneous to the patient, or a combination thereof.
34. The method according to claim 22, further comprising contacting unmodified cells with an expression vector encoding the expression of glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides the equivalent function, to produce the busulfan-resistant modified cells.
35. The method according to claim 34, wherein the expression vector is a viral vector or a non-viral vector.
36. The method according to claim 35, wherein the viral vector is a lentivirus or an adenovirus vector.
37. The method according to claim 34, wherein the expression vector is a retrovirus, a transposon, an episome expression vector, a modified RNA, a plasmid, or any combination thereof.
38. The method according to claim 22, wherein the at least one myeloablative dose of the chemotherapeutic agent is administered after the administration of the modified cells.
39. The method according to claim 22, wherein the at least one myeloablative dose of the chemotherapeutic agent is a myeloablative dose of busulfan.
40. The method according to claim 22, wherein the non-bone marrow-destroying chemotherapeutic agent is administered daily for at least one week, at least two weeks, at least three weeks, at least one month, at least two months, at least three months, at least four months, at least five months, or at least six months.
41. The method according to claim 22, further comprising refraining from administering the non-myeloablative dose of the chemotherapeutic agent for a period of time between the administration of the busulfan-resistant modified cells and the administration of at least one non-myeloablative dose of the chemotherapeutic agent.
42. The method according to claim 41, wherein the period is selected from the group consisting of approximately 3 days, approximately 7 days, approximately 10 days, and approximately 14 days.
43. The method according to claim 22, wherein more than 60%, 70%, 80%, 90%, 95%, or 100% of the bone marrow of the patient is replaced with the modified cells.
44. The method according to claim 22, wherein the patient is no longer in a myeloablative state and / or immunodeficiency state as a result of the administration of the at least one myeloablative dose of the chemotherapeutic agent.
45. The method according to claim 22, wherein the patient does not experience clinically significant anemia, neutropenia, thrombocytopenia, pancytopenia, decreased platelet count, decreased white blood cell count, or any combination thereof, or related symptoms.
46. The method according to claim 22, wherein a preconditioning step is performed before administration of the cells.
47. The method according to claim 46, wherein the preconditioning step is a non-myeloablative chemotherapy preconditioning step.
48. The method according to claim 22, wherein the modified cells are resistant to HIV infection.
49. The method according to claim 48, wherein the modified cells heterologously express at least one mutation of an HIV coreceptor that is resistant to HIV infection, one or more mutations of at least one HIV coreceptor, at least one HIV fusion inhibitor, a molecule that reduces the expression of an HIV coreceptor, or any combination thereof.
50. The method according to claim 48, wherein the modified cells heterologously express shCCR5, shCXCR4, a C-peptide fusion inhibitor, or any combination thereof.
51. The method according to claim 48, wherein the modified cells do not express the HIV coreceptor.
52. The method according to claim 48, wherein the modified cells do not express CCR5, CXCR4, or express CCR5-Δ32, or a combination thereof.
53. A heterogeneous nucleotide molecule encoding the expression of any other polypeptide in the GSH synthesis pathway that provides an equivalent function to the glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway, i) at least one HIV coreceptor mutation, at least one HIV co A cell comprising one or more mutations in the receptor, at least one HIV fusion inhibitor, a molecule that reduces the expression of the HIV co-receptor, or a heterogeneous nucleotide molecule encoding any combination thereof, and / or ii) one of the endogenous HIV co-receptor mutations or deletions.
54. The aforementioned cells, i. A heterogeneous nucleotide sequence encoding a molecule that reduces the expression of CCR5, ii. A heterogeneous nucleotide sequence that codes for reducing the expression of CXCR4, iii. Heterogeneous nucleotide sequences encoding the expression of the C peptide fusion inhibitor, The cell according to claim 53, comprising iv. or any combination thereof.
55. The cell according to claim 53, wherein the cell expresses shCCR5, shCXCR4, and / or a C-peptide fusion inhibitor, such as C44.
56. The cell according to claim 53, wherein the cell is modified to express the heterologous nucleotide sequence in conjunction with a nonviral gene transfer system.
57. The cell according to claim 56, wherein the nonviral gene transfer system is a transposon gene transfer system.
58. The cell according to claim 57, wherein the transposon gene transfer system is the Sleeping Beauty gene transfer system or the PiggyBac transposon gene transfer system.
59. A composition comprising one or more cells according to any one of claims 53 to 58.
60. A nucleic acid molecule encoding a heterologous glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function, and one heterologous nucleotide molecule encoding at least one HIV co-receptor mutation, one or more mutations of at least one HIV co-receptor, at least one HIV fusion inhibitor, a molecule that reduces the expression of HIV co-receptor, or any combination thereof.
61. The aforementioned heterogeneous nucleotide sequence, i. Molecules that reduce the expression of CCR5, ii. Molecules that reduce the expression of CXCR4, iii. The molecule encoding the expression of the C peptide fusion inhibitor, A nucleic acid molecule according to claim 60, which codes for any combination thereof.
62. The nucleic acid molecule according to claim 60, wherein the nucleic acid molecule encodes the expression of shCCR5, shCXCR4, and / or a C-peptide fusion inhibitor.
63. A vector comprising the nucleic acid molecule described in claim 60.
64. The vector according to claim 63, wherein the vector is a vector that can be used to produce a lentivirus.
65. The vector according to claim 63, wherein the vector is a lentiviral vector.
66. A method for treating HIV, wherein the subject is given glutamate cysteine A population of cells heterologously expressing one of the following: gauze (GCL) modifying factor subunits GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function; i) a heterologous nucleotide molecule encoding at least one HIV co-receptor mutation, one or more mutations of at least one HIV co-receptor, at least one HIV fusion inhibitor, a molecule that reduces the expression of the HIV co-receptor, or any combination thereof; The method comprising administering at least one myeloablative dose of a chemotherapeutic agent.
67. The method according to claim 66, wherein the cells express shCCR5, shCXCR4, and / or a C-peptide fusion inhibitor.
68. The method according to claim 66 or 67, wherein the cells heterologously express GCLM.
69. A method for expressing a target molecule in a subject, comprising administering to the subject a heterologous glutamate cysteine ligase (GCL) modifying factor subunit GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function, and cells heterologously expressing the target molecule, and administering a non-myeloablative dose of busfuran.
70. The method according to claim 69, wherein the cells are immune cells, stem cells, or CD34+ and / or CD4+ cells, or as otherwise provided herein.
71. The method according to claim 69, wherein the target molecule is a molecule that reduces the expression of CCR5, a molecule that reduces the expression of CXCR4, a molecule that encodes the expression of a C-peptide fusion inhibitor, or any combination thereof.
72. The method according to claim 71, wherein the target molecule that reduces the expression of CCR5 is shCCR5.
73. The method according to claim 69, wherein the target molecule is a wild-type protein that is deficient in A1AT deficiency, WAS, Flurer syndrome, Hunter syndrome, Pompe disease, Fabry disease, mucopolysaccharidosis, MPS 1 H / S (Flurer / Chaillet syndrome), MPS I H (Flurer disease), MPS II- (Hunter syndrome), MPS III A, B, C, and D (Sanfilippo syndrome), MPS I S (Chaillet syndrome), MPS IV A and B (Morcchio syndrome), MPS IX (hyaluronidase deficiency), MPS VII (Sly syndrome), MPS VI (Maroto-Lamy syndrome), lysosome storage disorder, and childhood cerebral adrenoleukodystrophy (cALD).
74. Isolated stem cells or immune cells containing heterologous glutamate cysteine ligase (GCL) modifying factor subunits GCLM, GCLC (GCL catalytic subunit), GCL (as a dimer or complete enzyme), or any other polypeptide in the GSH synthesis pathway that provides equivalent function.
75. The cell according to claim 74, wherein the cell comprises heterologous GCLM.
76. The cell according to claim 74 or 75, wherein the cell further comprises at least one additional heterologously expressed molecule of interest.
77. The aforementioned stem cells include fetal stem cells, umbilical cord blood-derived stem cells, hematopoietic stem cells (HSCs), and pluripotent stem cells. The method according to any one of claims 74 to 76, wherein the cells are (PSCs), induced PSCs (iPSCs), embryonic stem cells (ESCs) or cells derived therefrom, such as CD34+ cells, CD90+ cells, CD45+ cells, CD17+ cells, CD45RA- cells, or any combination thereof.
78. The cell according to any one of claims 74 to 76, wherein the immune cell is a T cell.
79. The cell according to claim 78, wherein the T cell further comprises a chimeric antigen receptor (CAR).
80. The cell according to any one of claims 74 to 79, wherein the heterologous GCLM protein comprises the amino acid sequence of SEQ ID NO: 1 or a variant thereof, or a sequence that is at least 90% identical to SEQ ID NO: 1, or substantially identical to the amino acid sequence of SEQ ID NO:
1.
81. The cells according to any one of claims 74 to 80, wherein the cells do not heterologously express any other proteins that confer chemotherapy resistance.
82. The cells according to any one of claims 74 to 81, wherein the cells do not heterologously express ALDH.
83. The cell according to any one of claims 74 to 81, wherein the cell heterologously expresses GCLM and endogenously expresses GCLM.
84. The pharmaceutical composition comprising the cells described in any one of claims 74 to 83 and a pharmaceutically acceptable buffer or excipient.
85. A method for treating cancer in a patient, the method comprising administering busulfan to the patient, wherein the patient is administered the cells described in claim 79.
86. The method according to claim 81, wherein the method comprises administering the cells according to claim 79 to the patient before administering the busulfan.
87. The method according to claim 85 or 86, wherein the busulfan is administered in a non-myeloablative dose.
88. The method according to claim 85 or 86, wherein the busulfan is administered in a myeloablative dose.
89. The method according to any one of claims 85 to 88, wherein the cancer is a hematological carcinoma or an osteosarcoma.
90. The method according to claim 89, wherein the blood cancer is leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, lymphoma, AIDS-related lymphoma, cutaneous T-cell lymphoma, Hodgkin lymphoma, Hodgkin lymphoma, mycosis fungoides, non-Hodgkin lymphoma, primary central nervous system lymphoma, Sézary syndrome, T-cell lymphoma, Waldenström macroglobulinemia, chronic myeloproliferative neoplasm, Langerhans cell histiocytosis, multiple myeloma, myelodysplastic syndrome, or myelodysplastic / myeloproliferative neoplasm.