IP Library › Granted Patent US 12,397,060
Granted Patent B2
US 12,397,060 · App. 17/423,849 · Granted Aug 26, 2025

pHLIP®-mediated intracellular delivery of immuno-stimulatory compounds

Inventors: Yana K. Reshetnyak (Saunderstown, RI); Oleg A. Andreev (Saunderstown, RI); Donald M. Engelman (New Haven, CT); Anna Moshnikova (Warwick, RI); John Deacon (Tolland, CT)
Assignees: University of Rhode Island Board of Trustees; Yale University
A61K47/64A61K9/0019A61K39/39A61K47/42
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Quick Facts
Patent No.
US 12,397,060
App. No.
17/423,849
Granted
Aug 26, 2025
Kind
B2
Abstract

The invention features a composition comprising an immuno-stimulatory compound and a pHLIP® peptide, e.g., an immunostimulatory compound that comprises a cyclic purine dinucleotide, which binds to a stimulator of interferon genes (STING) such as a cGAMP cyclic compound inside a cell.

Claims (82)

1. A composition comprising an immuno-stimulatory compound (ISC) and a pH low insertion peptide (pHLIP) peptide, wherein said ISC is selected from a group consisting of:

(i) a cyclic dinucleotide (CDN),

(ii) a cyclic purine dinucleotide,

(iii) a cyclic purine dinucleotide which binds to stimulator of interferon genes (STING),

(iv) a cGAMP,

(v) a 3′,5′-cyclic diadenylic acid (c-di-AMP), and

(vi) a cyclic diguanylate (c-di-GMP) cyclic compound.

2. The composition of claim 1 , comprising the following structure:

Peptide-Linker-ISC

wherein “Peptide” is a pHLIP peptide comprising the sequence

(SEQ ID NO: 212)

ADQDNPWRAYLDLLFPTDTLLLDLLWCA

or

(SEQ ID NO: 10)

ADDQNPWRAYLDLLFPTDTLLLDLLWCA

or

(SEQ ID NO: 213)

ADQDNPWRAYLDLLFPTDTLLLDLLWKA

or

(SEQ ID NO: 217)

ADDQNPWRAYLDLLFPTDTLLLDLLWKA

wherein “Linker” is a cleavable linker,

wherein “ISC” is a cyclic purine dinucleotide which binds to stimulator of interferon genes (STING), and

wherein each “—” is a covalent bond.

3. The composition of claim 2 , wherein said linker comprises a disulfide bond or an acid-labile bond.

4. The composition of claim 2 , wherein said linker is self-immolating.

5. The composition of claim 1 , further comprising a modulator of polarity.

6. The composition of claim 1 , wherein said composition comprises 2 or more pHLIP peptides.

7. The composition of claim 6 , comprising the following structure:

Peptide1-Link-Peptide2

wherein “Peptide1” is a first pHLIP peptide comprising the sequence

(SEQ ID NO: 1)

ADDQNPWRAYLDLLFPTDTLLLDLLWXA

or

(SEQ ID NO: 214)

ADQDNPWRAYLDLLFPTDTLLLDLLWXA,

wherein “Peptide2” is a second pHLIP peptide comprising the sequence

(SEQ ID NO: 1)

ADDQNPWRAYLDLLFPTDTLLLDLLWXA

or

(SEQ ID NO: 214)

ADQDNPWRAYLDLLFPTDTLLLDLLWXA,

wherein “X” indicates any amino acid residue, including a lysine (Lys), a cysteine (Cys), or an Azido-containing amino acid,

wherein “Link” is a polyethylene glycol linker, and

wherein each “—” is a covalent bond.

8. A method of augmenting an anti-tumor immune response, comprising administering to a subject a composition comprising an immuno-stimulatory compound and a pHLIP peptide, wherein said immuno-stimulatory compound is selected from the group consisting of:

(i) a cyclic dinucleotide (CDN),

(ii) a cyclic purine dinucleotide,

(iii) a cyclic purine dinucleotide which binds to stimulator of interferon genes (STING),

(iv) a cGAMP,

(v) a 3′,5′-cyclic diadenylic acid (c-di-AMP), and

(vi) a cyclic diguanylate (c-di-GMP) cyclic compound.

9. The method of claim 8 , wherein said subject comprises a solid tumor.

10. The method of claim 8 , wherein said composition is injected directly into a tumor mass.

11. The method of claim 8 , wherein said composition is systemically administered.

12. The method of claim 8 , wherein said immuno-stimulatory compound is delivered into the cytosols of cancer cells.

13. The method of claim 8 , wherein said immuno-stimulatory compound is delivered into the cytosol of a macrophage within the tumor microenvironment.

14. The method of claim 8 , wherein said immuno-stimulatory compound is delivered intracellularly to induce a biological effect.

15. The method of claim 14 , wherein the biological effect of said immuno-stimulatory compound delivered in the presence of said pHLIP is at least 20% greater than that delivered in the absence of said pHLIP.

16. The method of claim 8 , wherein said composition targets said immuno-stimulatory compound preferentially to a diseased tissue compared to a healthy tissue, thereby minimizing damage to said healthy tissue.

17. The method of claim 8 , wherein said composition selectively promotes intracellular delivery of said immuno-stimulatory compound to cells in diseased tissue.

18. The method of claim 17 , wherein said composition selectively promotes intracellular delivery of said immuno-stimulatory compound into a cancer cell.

19. The method of claim 8 , wherein said composition selectively promotes intracellular delivery of said immuno-stimulatory compound into macrophages in a tumor microenvironment.

20. The method of claim 8 , wherein said composition selectively promotes intracellular delivery of said immuno-stimulatory compound into a macrophage in a diseased tissue environment.

21. The method of claim 8 , wherein said pHLIP peptide comprises the amino acid sequence of

(SEQ ID NO: 10)

ADDQNPWRAYLDLLFPTDTLLLDLLWCA

or

(SEQ ID NO: 212)

ADQDNPWRAYLDLLFPTDTLLLDLLWCA.

22. The method of claim 8 , wherein the composition comprises the following structure:

Peptide-Link-ISC

wherein “Peptide” us a pHLIP peptide comprising an amino acid sequence of

(SEQ ID NO: 2)

AXDDQNPWRAYLDLLFPTDTLLLDLLWXA

or

(SEQ ID NO: 214)

ADQDNPWRAYLDLLFPTDTLLLDLLWXA,

wherein “X” is, selected from a lysine (Lys), a cysteine (Cys), or an Azido-containing amino acid,

wherein “Link” is a polyethylene glycol linker,

wherein “ISC” is said immuno-stimulatory compound, and

wherein each “—” is a covalent bond.

Assignments (3)
CONFIRMATORY LICENSE Recorded Dec 6, 2023
From: UNIVERSITY OF RHODE ISLAND
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 065791/0446 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 27, 2022
From: DEACON, JOHN; ENGELMAN, DONALD M.
To: YALE UNIVERSITY
Reel/Frame 059752/0009 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 27, 2022
From: RESHETNYAK, YANA K.; ANDREEV, OLEG A.; MOSHNIKOVA, ANNA
To: UNIVERSITY OF RHODE ISLAND BOARD OF TRUSTEES
Reel/Frame 059752/0036 →
Continuity (2)
Provisional Application 62797893 · Jan 28, 2019
Related Publication 20220088208A1 · Mar 24, 2022
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