Granted Patent
B2
US 12,357,653 · App. 16/772,678 · Granted Jul 15, 2025
Compositions comprising circular polyribonucleotides and uses thereof
Inventors:
Avak Kahvejian (Lexington, MA); Nicholas McCartney Plugis (Cambridge, MA); Alexandra Sophie De Boer (Cambridge, MA)
Assignee:
Flagship Pioneering Innovations VI, LLC
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Abstract
This invention relates generally to pharmaceutical compositions and preparations of circular polyribonucleotides and uses thereof.
Claims (78)
1. A method of delivering a circular polyribonucleotide to a subject, comprising administering to the subject a pharmaceutical composition for expression of a first chimeric antigen receptor (CAR) and a second CAR, the pharmaceutical composition comprising:
(a) a lipid nanoparticle carrier comprising a covalently closed polyribonucleotide that comprises, in the following order:
(i) a first internal ribosome entry site (IRES);
(ii) a first expression sequence that encodes the first CAR;
(iii) a second IRES; and
(iv) a second expression sequence that encodes the second CAR; and
(b) a pharmaceutically acceptable excipient.
2. The method of claim 1 , wherein the covalently closed polyribonucleotide lacks a 5′ cap.
3. The method of claim 1 , wherein the covalently closed polyribonucleotide is detectable in vivo for at least 7 days after administration of the composition to a human.
4. The method of claim 1 , wherein the covalently closed polyribonucleotide further comprises a miRNA target sequence or a miRNA binding site.
5. The method of claim 1 , wherein the covalently closed polyribonucleotide comprises, in the following order:
(i) the first IRES;
(ii) a first translation initiation sequence;
(iii) the first expression sequence that encodes the first CAR;
(iv) the second IRES;
(v) a second translation initiation sequence; and
(vi) the second expression sequence that encodes the second CAR.
6. The method of claim 1 , wherein the covalently closed polyribonucleotide comprises, in the following order:
(i) a first spacer sequence;
(ii) the first IRES;
(iii) the first expression sequence that encodes the first CAR;
(iv) a second spacer;
(v) the second IRES; and
(vi) the second expression sequence that encodes the second CAR.
7. The method of claim 6 , wherein the covalently closed polyribonucleotide comprises, in the following order:
(i) the first spacer sequence;
(ii) the first IRES;
(iii) the first expression sequence that encodes the first CAR;
(iv) the second spacer;
(v) the second IRES;
(vi) the second expression sequence that encodes the second CAR; and
(vii) a third spacer.
8. The method of claim 1 , wherein the covalently closed polyribonucleotide comprises, in the following order:
(i) a first spacer sequence;
(ii) the first IRES;
(iii) the first expression sequence that encodes the first CAR;
(iv) the second IRES;
(v) the second expression sequence that encodes the second CAR; and
(vi) a second spacer.
9. A method of delivering a circular polyribonucleotide to a subject, comprising administering to the subject a pharmaceutical composition for expression of a first CAR and a second CAR, the pharmaceutical composition comprising:
(a) a lipid nanoparticle carrier comprising a covalently closed polyribonucleotide that comprises, in the following order:
(i) an IRES;
(ii) a first expression sequence that encodes the first CAR;
(iii) a stagger element;
(iv) a second expression sequence that encodes the second CAR; and
(b) a pharmaceutically acceptable excipient.
10. The method of claim 9 , wherein the covalently closed polyribonucleotide lacks a 5′ cap.
11. The method of claim 9 , wherein the covalently closed polyribonucleotide is detectable in vivo for at least 7 days after administration of the composition to a human.
12. The method of claim 9 , wherein the covalently closed polyribonucleotide further comprises a miRNA target sequence or a miRNA binding site.
13. The method of claim 9 , wherein the covalently closed polyribonucleotide comprises, in the following order:
(i) the IRES;
(ii) a first translation initiation sequence;
(iii) the first expression sequence that encodes the first CAR;
(iv) the stagger element;
(v) a second translation initiation sequence; and
(vi) the second expression sequence that encodes the second CAR.
14. The method of claim 9 , wherein the covalently closed polyribonucleotide comprises, in the following order:
(i) a first spacer sequence;
(ii) the IRES;
(iii) the first expression sequence that encodes the first CAR;
(iv) a second spacer;
(v) the stagger element; and
(vi) the second expression sequence that encodes the second CAR.
15. The method of claim 14 , wherein the covalently closed polyribonucleotide comprises, in the following order:
(i) the first spacer sequence;
(ii) the IRES;
(iii) the first expression sequence that encodes the first CAR;
(iv) the second spacer;
(v) the stagger element;
(vi) the second expression sequence that encodes the second CAR; and
(vii) a third spacer.
16. The method of claim 9 , wherein the covalently closed polyribonucleotide comprises, in the following order:
(i) a first spacer sequence;
(ii) the IRES;
(iii) the first expression sequence that encodes the first CAR;
(iv) the stagger element;
(v) the second expression sequence that encodes the second CAR; and
(vi) a second spacer.
Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jan 8, 2021
From: KAHVEJIAN, AVAK; PLUGIS, NICHOLAS MCCARTNEY; DE BOER, ALEXANDRA SOPHIE; WEINSTEIN, ERICA GABRIELLE; TROUSIL, SEBASTIAN
To: FLAGSHIP PIONEERING, INC.
Reel/Frame 054852/0683 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jan 8, 2021
From: STEWART, MORAG HELEN; CIFUENTES-ROJAS, CATHERINE; PAEK, KI YOUNG
To: VL50, INC.
Reel/Frame 054852/0692 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jan 8, 2021
From: VL50, INC.
To: FLAGSHIP PIONEERING, INC.
Reel/Frame 054853/0467 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jan 8, 2021
From: FLAGSHIP PIONEERING, INC.
To: FLAGSHIP PIONEERING INNOVATIONS VI, LLC
Reel/Frame 054853/0490 →
Continuity (3)
Provisional Application
62676688
· May 25, 2018
Provisional Application
62599547
· Dec 15, 2017
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