IP Library › Granted Patent US 12,202,863
Granted Patent B2
US 12,202,863 · App. 17/343,022 · Granted Jan 21, 2025

Chimeric antigen receptor (CAR) modulation

Inventors: John Ngo (Cambridge, MA); Wilson Wong (Brookline, MA); Meng Lai Nicole Wong (Watertown, MA); Huishan Li (Brookline, MA); Elliot P. Tague (Brookline, MA)
Assignee: TRUSTEES OF BOSTON UNIVERSITY
C07K14/1833C07K14/811C07K2319/02C07K2319/03C07K2319/50C07K2319/80
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Quick Facts
Patent No.
US 12,202,863
App. No.
17/343,022
Granted
Jan 21, 2025
Kind
B2
Abstract

The technology described herein is directed to CAR polypeptides and systems comprising repressible proteases. In combination with a specific protease inhibitor, the activity of said CAR polypeptides and systems and cells comprising them can be modulated. Also described herein are methods of using said CAR polypeptides and systems, for example to treat various diseases and disorders.

Claims (55)

1. A polypeptide system comprising:

a) a first polypeptide comprising:

(i) an extracellular binding domain;

(ii) a transmembrane domain; and

(iii) a reader domain comprising a danoprevir/NS3 complex reader domain (DNCR) or a grazoprevir/NS3 reader complex (GNCR) domain; and

b) a second polypeptide comprising:

(i) a repressible protease comprising hepatitis C virus (HCV) nonstructural protein 3 (NS3) genotype 1B comprising a sequence selected from the group consisting of SEQ ID NO: 99, SEQ ID NO: 130, and SEQ ID NO: 180; and

(ii) at least one intracellular signaling domain.

2. The polypeptide system of claim 1 , wherein the reader domain specifically binds to the repressible protease in the presence of a specific protease inhibitor.

3. The polypeptide system of claim 1 , wherein the first polypeptide does not comprise a protease cleavage site.

4. The polypeptide system of claim 1 , wherein the first polypeptide further comprises at least one intracellular signaling domain.

5. The polypeptide system of claim 4 , wherein the at least one intracellular signaling domain is selected from the group consisting of: TCRC; FcRy; FcRp; CD3zeta; CD3y; CD35; CD3s; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM-1); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; ZAP70; and 41BB.

6. The polypeptide system of claim 1 , wherein the first polypeptide comprises:

a) a polypeptide comprising from the N-terminus to the C-terminus:

i) an extracellular binding domain;

ii) a transmembrane domain;

iii) a first intracellular signaling domain;

iv) a second intracellular signaling domain; and

v) a reader domain;

b) a polypeptide comprising from the N-terminus to the C-terminus:

i) an extracellular binding domain;

ii) a transmembrane domain;

iii) a single intracellular signaling domain; and

iv) and a reader domain; or

c) a polypeptide comprising residues 1-626 of SEQ ID NO: 127, residues 1-630 of SEQ ID NO: 128, or a sequence that is at least 70% identical to residues 1-626 of SEQ ID NO: 127 or residues 1-630 of SEQ ID NO: 128, that maintains the same function.

7. The polypeptide system of claim 1 , wherein the at least one intracellular signaling domain of the second polypeptide is selected from the group consisting of: TCRC; FcRy; FcRp; CD3zeta; CD3y; CD35; CD3s; CD3C; CD22; CD79a; CD79b; CD66d; CARD11; CD2; CD7; CD27; CD28; CD30; CD40; CD54 (ICAM-1); CD83; CD134 (OX40); CD137 (4-1BB); CD150 (SLAMF1); CD152 (CTLA4); CD223 (LAG3); CD270 (HVEM); CD273 (PD-L2); CD274 (PD-L1); CD278 (ICOS); DAP10; LAT; KD2C SLP76; TRIM; ZAP70; and 41BB.

8. The polypeptide system of claim 1 , wherein the repressible protease is catalytically dead.

9. The polypeptide system of claim 1 , wherein the repressible protease of the second polypeptide is located:

i) between a transmembrane domain and the at least one intracellular signaling domain;

ii) between a first intracellular signaling domain and a second intracellular signaling domain; or

iii) at the C terminus of the polypeptide.

10. The polypeptide system of claim 1 , wherein the second polypeptide further comprises a cofactor for the repressible protease.

11. The polypeptide system of claim 10 , wherein the cofactor is an HSV NS4A domain.

12. The polypeptide system of claim 1 , wherein the second polypeptide does not comprise a protease cleavage site.

13. The polypeptide system of claim 1 , wherein the second polypeptide is in combination with a protease inhibitor bound to the repressible protease.

14. The polypeptide system of claim 13 , wherein the protease inhibitor is selected from grazoprevir (GZV), danoprevir, simeprevir, asunaprevir, ciluprevir, boceprevir, sovaprevir, paritaprevir, ombitasvir, paritaprevir, ritonavir, dasabuvir, and telaprevir.

15. The polypeptide system of claim 13 , wherein the reader domain of the first polypeptide specifically binds to the repressible protease of the second polypeptide in the presence of the protease inhibitor specific to the reader domain; wherein the reader domain of the first polypeptide does not specifically bind to the repressible protease of the second polypeptide in the absence of the protease inhibitor specific to the reader domain.

16. The polypeptide system of claim 1 , wherein the second polypeptide comprises:

a) a polypeptide comprising from the N-terminus to the C-terminus:

i) an extracellular domain;

ii) a transmembrane domain;

iii) at least one intracellular signaling domain;

iv) a repressible protease; and

v) at least one intracellular signaling domain;

b) a polypeptide comprising from the N-terminus to the C-terminus:

i) an extracellular domain;

ii) a transmembrane domain;

iii) a first intracellular signaling domain;

iv) a repressible protease; and

v) a second intracellular signaling domain; or

c) a polypeptide comprising one of SEQ ID NO: 86-89 or residues 648-1303 of SEQ ID NO: 127, or residues 652-1307 of SEQ ID NO: 128, or a sequence that is at least 70% identical to one of SEQ ID NO: 86-89, residues 648-1303 of SEQ ID NO: 127, or residues 652-1307 of SEQ ID NO: 128 that maintains the same function.

17. The polypeptide system of claim 1 , wherein the first polypeptide and second polypeptide are physically linked to one another; flank a self-cleaving peptide domain; and/or comprise one of SEQ ID NOs: 127-128 or a sequence that is at least 70% identical to SEQ ID NOs: 127-128 that maintains the same function.

18. The polypeptide system of claim 1 , wherein the repressible protease comprises SEQ ID NO: 99.

19. The polypeptide system of claim 1 , wherein the repressible protease comprises SEQ ID NO: 130 or SEQ ID NO: 180.

20. The polypeptide system of claim 1 , wherein the reader domain is a danoprevir/NS3 complex reader (DNCR) domain.

Assignments (2)
CORRECTIVE ASSIGNMENT TO CORRECT THE THIRD ASSIGNOR'S NAME PREVIOUSLY RECORDED AT REEL: 057020 FRAME: 0246. ASSIGNOR(S) HEREBY CONFIRMS THE ASSIGNMENT. Recorded Aug 16, 2021
From: NGO, JOHN; WONG, WILSON; WONG, MENG LAI NICOLE; LI, HUISHAN; TAGUE, ELLIOT P.
To: TRUSTEES OF BOSTON UNIVERSITY
Reel/Frame 057193/0354 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 29, 2021
From: NGO, JOHN; WONG, WILSON; NICOLE WONG, MENG LAI; LI, HUISHAN; TAGUE, ELLIOT P.
To: TRUSTEES OF BOSTON UNIVERSITY
Reel/Frame 057020/0246 →
Continuity (3)
Division 16832751 · Mar 27, 2020
Provisional Application 62826139 · Mar 29, 2019
Related Publication 20220098246A1 · Mar 31, 2022
References Cited (36)
US 6333186B1 · Wittekind et al. · 2001 [cited by applicant]
US 7208309B2 · Kukolj et al. · 2007 [cited by applicant]
US 7494660B2 · Lin et al. · 2009 [cited by applicant]
US 9889160B2 · Jantz et al. · 2018 [cited by applicant]
US 10137180B2 · Wandless et al. · 2018 [cited by applicant]
US 20020106642A1 · Wittekind et al. · 2002 [cited by applicant]
US 20160311907A1 · Brogdon · 2016 [cited by examiner]
US 20170157176A1 · Wang et al. · 2017 [cited by applicant]
US 20180011096A1 · Chien et al. · 2018 [cited by applicant]
US 20180179522A1 · Buckley et al. · 2018 [cited by applicant]
WO 2016210447A1 · 2016 [cited by applicant]
WO 2018206791A1 · 2018 [cited by applicant]
WO 2019118518A2 · 2019 [cited by applicant]
WO 2020118076A1 · 2020 [cited by applicant]
Seet et. al. (Nat. Rev. Mol Cell Biol. 7:473-483. (2006)) (Year: 2006). [cited by examiner]
Lemmon et. al. (Cell. 141(7):1117-34. (2010)) (Year: 2010). [cited by examiner]
Wu et al. (Science 350(6258): 293(Summary) & 1-10 (2015) (Year: 2015). [cited by examiner]
Foight et. al. Nat Biotechnol 37:1209-1216 (2019) (Year: 2019). [cited by examiner]
Ku et. al. (Journal of Microbiology. 54(7):1835-1841 (2016)) (Year: 2016). [cited by examiner]
Morikawa et. al. (Journal of Viral Hepatitis. 18:305-315. (2011)) (Year: 2011). [cited by examiner]
Han et al. “Masked chimeric antigen receptor for tumor-specific activation.” Molecular Therapy 25.1: 274-284 (2017). [cited by applicant]
Tague. et al. “Chemogenetic control of gene expression and cell signaling with antiviral drugs.” Nature methods 15.7: 519-522 (2018). [cited by applicant]
Banaszynski et al., “A rapid, reversible, and tunable method to regulate protein function in living cells using synthetic small molecules.” Cell 126(5): 995-1004 (2006). [cited by applicant]
Bonger et al., “Small molecule displacement of a cryptic degron causes conditional protein degradation” Nat. Biol. Chem. (2011) 7(8):531-537. [cited by applicant]
Cunningham-Bryant et al., “A chemically disrupted proximity system for controlling dynamic cellular processes.” Journal of the American Chemical Society 141(8): 3352-3355 (2019). [cited by applicant]
Foight et al., “Multi-input chemical control of protein dimerization for programming graded cellular responses.” Nature biotechnology 37(10): 1209-1216 (2019). [cited by applicant]
Gonda et al., “Universality and Structure of the N-end Rule”, The Journal of Biological Chemistry 264(28): 16700-16712 (1989). [cited by applicant]
Kliemann et al., “Polymorphisms and resistance mutations of hepatitis C virus on sequences in the European hepatitis C virus database.” World journal of gastroenterology 22(40): 8910-8917 (2016). [cited by applicant]
Kugler et al., “High affinity peptide inhibitors of the hepatitis C virus NS3-4A protease refractory to common resistant mutants.” Journal of Biological Chemistry 287(46): 39224-39232 (2012). [cited by applicant]
Lin “HCV NS3-4A Serine Protease” Hepatitis C Viruses: Genomes and Molecular Biology, Chapter 6: 163-206 (2006). [cited by applicant]
Mccauley et al., “Hepatitis C virus NS3/4a protease inhibitors.” Current opinion in pharmacology 30: 84-92 (2016). [cited by applicant]
Schrader et al., “Making it easier to regulate protein stability.” Chemistry & biology 17(9): 917-918 (2010). [cited by applicant]
Sheridan et al., “Selectable one-step PCR-mediated integration of a degron for rapid depletion of endogenous human proteins.” BioTechniques 60(2): 69-74 (2016). [cited by applicant]
Sun et al., “Analysis of Naturally Occurring Resistance-Associated Variants to NS3/4A Protein Inhibitors, NS5A Protein Inhibitors, and NS5B Polymerase Inhibitors in Patients With Chronic Hepatitis C.” Gene Expression Th… [cited by applicant]
Forst “Recognition of mono-ADP-ribosylated ARTD10 substrates by ARTD8 macrodomains and acetylation of ARTD10.” Thesis Diss. Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen University, (2013). [cited by applicant]
Schmidt “Design of an allosterically controlled serpin-based drug shuttle for the delivery of doxycycline and doxorubicin.” Thesis Diss. Friedrich-Alexander-University Erlangen-Nuremberg, (2018). [cited by applicant]