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US 2026/0226167 A1Published application
In vivo t cell gene editing
AssigneeGenzyme Corporation
InventorsChristopher BORGES, Viktor T. LEMGART, Matthew MACKEY, Shu-Wei WU
Life of this patent
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Abstract
The present disclosure relates generally to compositions and methods for in vivo gene therapy, and more specifically to delivering mRNA encoding anti-CD19 chimeric antigen receptors (CARs) to immune cells (e.g., T cells, such as CD8+ T cells) in vivo.
Claims32
1:
- A polypeptide comprising an anti-CD19 chimeric antigen receptor (anti-CD19 CAR), wherein the anti-CD19 CAR comprises:
- (a) an anti-CD19 antigen-binding domain, wherein the anti-CD19 antigen-binding domain is an anti-CD19 single domain antibody (anti-CD19 sdAb) domain comprising an anti-CD19 VHH comprising:
- (i) an H-CDR1 comprising the amino acid sequence of SEQ ID NO:427, an H-CDR2 comprising the amino acid sequence of SEQ ID NO:428, and an H-CDR3 comprising the amino acid sequence of SEQ ID NO:429, or
- (ii) an H-CDR1 comprising the amino acid sequence of SEQ ID NO:475, an H-CDR2 comprising the amino acid sequence of SEQ ID NO:476, and an H-CDR3 comprising the amino acid sequence of SEQ ID NO:477;
- (b) a hinge domain,
- (c) a transmembrane domain,
- (d) an intracellular signaling domain, and
- (e) optionally a co-stimulatory domain.
5 more elements
2:
- The polypeptide of claim 1, wherein the anti-CD19 sdAb comprises a VHH comprising the amino acid sequence of SEQ ID NO:433 or 481, or an amino acid sequence comprising at least about 80% sequence identity to the amino acid sequence of SEQ ID NO:433 or 481.
3:
- The polypeptide of claim 1, wherein the hinge domain is derived from a protein selected from the group consisting of CD28, CD5, CD9, CD2, CD27, CD28, IgG, IgK, and CD8α.
4:
- The polypeptide of claim 3, wherein in the hinge domain is derived from CD28 or CD8α.
5:
- The polypeptide of claim 1, wherein the transmembrane domain is derived from a protein selected from the group consisting of CD28, CD3ε, CD45, CD4, CD5, CD8α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD147, 4-1BB (CD137), CD152, PD-1, CD2, CD27, CD11b, CD11d, CD11a, CD11c, CD84, and CD86.
6:
- The polypeptide of claim 5, wherein in the transmembrane domain is derived from CD28 or CD8α.
7:
- The polypeptide of claim 1, wherein the hinge domain and the transmembrane domain are derived from the same protein.
8:
- The polypeptide of claim 7, wherein the hinge domain and the transmembrane domain are derived from CD28 and comprises the amino acid sequence of SEQ ID NO:431 or 434.
(canceled)
The polypeptide of claim 8, wherein the intracellular signaling domain is derived from CD32, wherein the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO:544.
(canceled)
12:
- The polypeptide of claim 1, wherein the anti-CD19 CAR comprises a co-stimulatory domain derived from a protein selected from the group consisting of CD27, CD28, CD137 (4-1BB), OX40, CD40, PD-1, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, TNFRSF9, TNFRSF4, TNFRSF8, CD40LG, ITGB2, KLRC2, TNFRSF18, TNFRSF14, HAVCR1, LGALS9, CD83, ligands of CD83, and any combination thereof.
13:
- The polypeptide of claim 12, wherein the co-stimulatory domain is derived from CD137 (4-1BB) or CD28.
14:
- The polypeptide of claim 13, wherein the co-stimulatory domain comprises the amino acid sequence of SEQ ID NO:432 or 543.
15:
- The polypeptide of claim 1, further comprising a signal peptide sequence that comprises a signal peptide, wherein the peptide sequence is connected to the N-terminus of the anti-CD19 CAR.
16:
- The polypeptide of claim 15, wherein the signal peptide sequence is derived from a CD8α propeptide.
17:
- The polypeptide of claim 15, wherein the signal peptide sequence comprises the amino acid sequence of SEQ ID NO:515.
(canceled)
19:
- The polypeptide of claim 1, wherein the anti-CD19 CAR comprises the amino acid sequence of any one of SEQ ID NOs:49-51 and 435-437.
20:
- An isolated nucleic acid encoding the polypeptide of claim 1.
21:
- An isolated nucleic acid comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:46-48, 114, 145, 441-458, 482-511, and 570-575.
22-28. (canceled)
29:
- A lipid nanoparticle (LNP) comprising:
- (a) a lipid-immune cell targeting group conjugate comprising the compound of Formula (II):
- [Lipid]-[optional linker]-[binding domain], wherein the binding domain specifically binds to human CD8α,
- (b) an ionizable cationic lipid, and
- (c) a nucleic acid encoding the polypeptide of claim 1, wherein the nucleic acid is encapsulated in the LNP.
2 more elements
30:
- A lipid nanoparticle (LNP) comprising:
- (a) a lipid-immune cell targeting group conjugate comprising the compound of Formula (II):
- [Lipid]-[optional linker]-[binding domain], wherein the binding domain specifically binds to human CD8α,
- (b) an ionizable cationic lipid, and
- (c) a nucleic acid comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:46-48, 114, 145, 441-458, 482-511, and 570-575, wherein the nucleic acid is encapsulated in the LNP.
2 more elements
31:
- The LNP of claim 29, wherein:
- a. the binding domain is an immunoglobulin single variable domain (ISVD) that specifically binds to human CD8α, wherein the ISVD comprises a complementarity-determining region 1 (CDR1), a CDR2, and a CDR3 of an ISVD having the sequence selected from the group consisting of SEQ ID NOs:
- 160-179;
- or
- b. the binding domain is an immunoglobulin single variable domain (ISVD) that specifically binds to human CD8α, wherein the ISVD specifically binding to CD8α comprises a CDR1, a CDR2, and a CDR3 according to the Abm CDR definition,
- wherein CDR1 is chosen from the group consisting of:
- (i) SEQ ID NO:
- 244;
- and
- (ii) amino acid sequences that have 3, 2, or 1 amino acid difference with at least one of the amino acid sequences of SEQ ID NO:
- 244;
- wherein CDR2 is chosen from the group consisting of:
- (iii) SEQ ID NO:
- 246;
- and
- (iv) amino acid sequences that have 3, 2, or 1 amino acid difference with at least one of the amino acid sequences of SEQ ID NO:
- 246;
- and
- wherein CDR3 is chosen from the group consisting of:
- (v) SEQ ID NO:
- 248;
- and
- amino acid sequences that have 3, 2, or 1 amino acid difference with at least one of the amino acid sequences of SEQ ID NO:
- 248.
21 more elements
32:
- The LNP of claim 29, wherein the binding domain specifically binding to human CD8α is covalently coupled to the Lipid in Formula (II) via a linker comprising polyethylene glycol (PEG).
33-60. (canceled)
61:
- The LNP of claim 29, wherein the ionizable cationic lipid comprises a compound of Formula (I):
- or a salt thereof, or both, wherein:
- R1, R2, and R3 are each independently a bond or C1-3 alkylene;
- R1A, R2A, and R3A are each independently a bond or C1-10 alkylene;
- R1A1, R1A2, R1A3, R2A1, R2A2, R2A3, R3A1, R3A2, and R3A3 are each independently H, C1-20 alkyl, C1-20 alkenyl, —(CH2)0-10C(O)ORa1, or —(CH2)0-10OC(O)Ra2;
- Ra1 and Ra2 are each independently C1-20 alkyl or C1-20 alkenyl;
- R3B is
- R3B1 is C1-6 alkylene;
- and
- R3B2 and R3B3 are each independently H, unsubstituted C1-6 alkyl, or C1-6 alkyl substituted with 1 or 2-OH.
7 more elements
62-86. (canceled)
87:
- A method of modulating immune response in a human subject, comprising administering the LNP of claim 29 to the human subject.
88:
- A method of treating a B cell malignancy in a human subject comprising administering the LNP of claim 29 to the human subject.
89-102. (canceled)
Dependencies are inferred from the claim text.