Chimeric antigen receptors to control HIV infection

Patent metadata

Field	Value
Publication number	US-10246505-B2
Application number	US-201415039384-A
Country	US
Kind code	B2
Filing date	Nov 25, 2014
Priority date	Nov 25, 2013
Publication date	Apr 2, 2019
Grant date	Apr 2, 2019

Abstract

The present disclosure is directed to novel multispecific chimeric antigen receptor (CAR) proteins and DNA sequences encoding these proteins. The CARs comprise at least two extracellular domains fused, via a transmembrane domain to a cytoplasmic signaling domain comprising two signaling domains. The disclosure further relates to nucleic acids encoding the novel CARs, to host cells expressing the novel CARs, and to methods of using the CARs to co-stimulate effector functions in the cells and for using cells expressing the receptors for treatment of disease and viral infections. The disclosure also relates to methods of generating a recombinant T cell with reduced susceptibility to HIV infection.

First claim

We claim:

1. A multispecific chimeric antigen receptor protein, comprising:
an N-terminal extracellular targeting segment comprising:
a first targeting domain comprising a CD4 domain that binds to HIV Env, wherein the CD4 domain comprises the D1 and D2 extracellular domains of CD4 comprising amino acids 1-208 of SEQ ID NO: 53; and
a second targeting domain comprising a carbohydrate recognition domain (CRD) from a human C-type lectin that binds to HIV Env, wherein the CRD is selected from the group consisting of a CRD from mannose-binding lectin 2 (MBL2) comprising amino acids 214-330 of SEQ ID NO: 53 and a CRD from Langerin comprising amino acids 214-354 of SEQ ID NO: 51;
wherein the first and second targeting domains bind to different sites on HIV Env, and wherein the multispecific chimeric antigen receptor protein binds to HIV Env.

2. The multispecific chimeric antigen receptor protein of claim 1 , comprising a linker connecting the first targeting domain to the second targeting domain.

3. The multispecific chimeric antigen receptor protein of claim 1 , comprising an amino acid sequence selected from the group consisting of the amino acid sequence of SEQ ID NO: 51, and the amino acid sequence of SEQ ID NO: 53.

4. The multispecific chimeric antigen receptor protein of claim 1 , further comprising:
a linker connecting the extracellular targeting segment to a transmembrane domain;
the transmembrane domain; and
an intracellular region, comprising:
a cytoplasmic co-stimulatory signaling domain; and
a cytoplasmic effector function signaling domain,

wherein the transmembrane domain is linked or fused to the intracellular region.

5. The multispecific chimeric antigen receptor protein of claim 4 , wherein:
the transmembrane domain is from CD28;
the cytoplasmic co-stimulatory signaling domain is from CD28; and/or
the cytoplasmic effector function signaling domain is from CD3 zeta.

6. The multispecific chimeric antigen receptor protein of claim 2 , wherein the linker connecting the first targeting domain to the second targeting domain is no more than 20 amino acids long.

7. The multispecific chimeric antigen receptor protein of claim 6 , wherein the linker connecting the first targeting domain to the second targeting domain is ten amino acids long.

8. The multispecific chimeric antigen receptor protein of claim 1 , comprising:
the N-terminal extracellular targeting segment comprising:
the first targeting domain; and
the second targeting domain, wherein the second targeting domain is a MBL2 domain;

a transmembrane region from CD28; and
an intracellular region, comprising:
a cytoplasmic co-stimulatory signaling domain from CD28; and
a cytoplasmic effector function signaling domain from CD3 zeta,

wherein the N-terminal extracellular targeting segment is linked to the transmembrane region and the transmembrane region is linked or fused to the intracellular region.

9. The multispecific chimeric antigen receptor protein of claim 8 , wherein:
the N-terminal extracellular targeting segment comprises the amino acid sequence of SEQ ID NO: 53;
the transmembrane region from CD28 comprises the amino acid sequence of SEQ ID NO: 55;
the cytoplasmic co-stimulatory signaling domain from CD28 comprises the amino acid sequence of SEQ ID NO: 58; and
the cytoplasmic effector function signaling domain from CD3 zeta comprises the amino acid sequence of SEQ ID NO: 56.

10. A T cell comprising the multispecific chimeric antigen receptor protein of claim 1 .

11. The T cell of claim 10 , wherein the T cell is a CD8 + T cell and/or a CD4 + T cell.

12. The T cell of claim 10 , wherein the T cell is not susceptible to HIV infection.

13. A composition, comprising the T cell of claim 10 and a carrier.

14. A method of killing HIV-infected cells, comprising:
contacting the composition of claim 13 with an HIV-infected cell expressing gp120, thereby killing the HIV-infected cells.

15. A method of reducing the level of HIV infected cells in a subject infected with HIV, comprising:
administering to the subject a therapeutically effective amount of the composition of claim 13 , thereby treating the subject infected with HIV.

16. The method of claim 15 , wherein the T cell in the composition is a T cell that is not susceptible to HIV infection.

17. A method of treating a subject with an HIV infection, comprising: administering to the subject a therapeutically effective amount of the composition of claim 13 under conditions sufficient to form an immune complex of the extracellular targeting segment on the chimeric antigen receptor protein and the extracellular domain of an HIV Env protein in the subject.

Assignees

• Us Health

Inventors

• Berger Edward A
• Ghanem Mustafa H
• Dey Barna

Classifications

• A61K48/005

  characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title

• C07K14/7051

  T-cell receptor (TcR)-CD3 complex · CPC title

• C07K14/70564

  Selectins, e.g. CD62 · CPC title

• C07K2319/33

  fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies · CPC title