Granted Patent B2

US 12,570,699 · App. 17/132,607 · Granted Mar 10, 2026

Inventors: Arun Srivastava (Gainesville, FL); Li Zhong (Boxborough, MA); Sergei Zolotukhin (Gainesville, FL); George Vladimirovich Aslanidi (Austin, MN); Mavis Agbandje-McKenna (Gainesville, FL); Kim M. Van Vliet (Gainesville, FL); Chen Ling (Gainesville, FL)

Assignee: University of Florida Research Foundation, Incorporated

C07K14/005A61K35/76A61K48/0008A61K48/005A61K48/0091C12N7/00C12N15/86C12N15/861C12N15/8645A61K2039/5158C12N2750/14122C12N2750/14132C12N2750/14142C12N2750/14143C12N2750/14145C12N2750/14171C12N2810/6027

Abstract

Claims (34)

1 . A modified AAV capsid protein comprising any one of the following combinations of non-native amino acid substitutions:

(a) Y705F, Y731F, and any one of the following:

(i) T492V;

(ii) K533R;

(iii) S663V;

(iv) S663V, and T492V; or

(v) S663V, T492V, and K533R; or

(b) S663V, T492V and K533R;

of the wild-type AAV3 capsid protein as set forth in SEQ ID NO:3, or at the equivalent surface-exposed amino acid residues in any one of the corresponding wild-type AAV1, AAV2, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, or AAV10 capsid proteins, as set forth in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10, respectively, or in any combination thereof, or

(c) Y444F and T550V;

(d) S458V and S492V, but not S662V; or

(e) Y444F, Y500F, Y730F and S458V;

of the wild-type AAV2 capsid protein as set forth in SEQ ID NO:2, or at the equivalent surface-exposed amino acid residues in any one of the corresponding wild-type AAV1, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, or AAV10 capsid proteins, as set forth in SEQ ID NO: 1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10, respectively, or in any combination thereof.

2 . An isolated nucleic acid segment that encodes the modified AAV capsid protein of claim 1 .

3 . A recombinant adeno-associated viral (rAAV) particle comprising the modified AAV capsid protein of claim 1 .

4 . The rAAV particle of claim 3 , further comprising a nucleic acid segment that encodes a diagnostic or therapeutic agent operably linked to a promoter capable of expressing the nucleic acid segment in a suitable host cell comprising the particle.

5 . The rAAV particle of claim 4 , wherein the nucleic acid segment further comprises an enhancer, a post-transcriptional regulatory sequence, a polyadenylation signal, or any combination thereof, operably linked to the nucleic acid segment.

6 . The rAAV particle of claim 4 , wherein the promoter is a heterologous promoter, a tissue-specific promoter, a cell-specific promoter, a constitutive promoter, an inducible promoter, or any combination thereof.

7 . The rAAV particle of claim 4 , wherein the promoter is a liver-specific promoter, a tumor cell-specific promoter, or a combination thereof.

8 . The rAAV particle of claim 4 , wherein the nucleic acid segment expresses or encodes a polypeptide, a peptide, a ribozyme, a peptide nucleic acid, an siRNA, an RNAi, an antisense oligonucleotide, an antisense polynucleotide, an antibody, an antigen binding fragment, or any combination thereof.

9 . The rAAV particle of claim 4 , wherein the nucleic acid segment encodes a chemotherapeutic agent.

10 . The rAAV particle of claim 4 , wherein the diagnostic or therapeutic agent is an agonist, an antagonist, an anti-apoptosis factor, an inhibitor, a receptor, a cytokine, a cytotoxin, an erythropoietic agent, a glycoprotein, a growth factor, a growth factor receptor, a hormone, a hormone receptor, an interferon, an interleukin, an interleukin receptor, a nerve growth factor, a neuroactive peptide, a neuroactive peptide receptor, a protease, a protease inhibitor, a protein decarboxylase, a protein kinase, a protein kinase inhibitor, an enzyme, a receptor binding protein, a transport protein or an inhibitor thereof, a serotonin receptor, or an uptake inhibitor thereof, a serpin, a serpin receptor, a tumor suppressor, a cytotoxic agent, a cytostatic agent, an anti-inflammatory agent, or any combination thereof.

11 . The modified AAV capsid protein of claim 1 , wherein the capsid protein is an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, or AAV10 capsid protein.

12 . The modified AAV capsid protein of claim 1 , wherein the capsid protein is an AAV2 capsid protein.

13 . The modified AAV capsid protein of claim 1 , wherein the capsid protein is an AAV3 capsid protein.

14 . An isolated mammalian host cell comprising the modified AAV capsid protein of claim 1 or a nucleic acid encoding the modified AAV capsid protein.

15 . The isolated mammalian host cell of claim 14 , wherein the host cell is a stem cell, a hematopoietic cell, a blood cell, a neural cell, a retinal cell, an epithelial cell, an endothelial cell, a pancreatic cell, a cancer cell, a muscle cell, a vascular cell, a diaphragm cell, a stomach cell, a liver cell, a tumor cell, or a CD34+ cell.

16 . A composition comprising:

(i) a viral particle that comprises the modified AAV capsid protein of claim 1 and a nucleic acid segment that encodes a selected diagnostic or therapeutic agent; and

(ii) a pharmaceutically-acceptable buffer, diluent, or excipient.

17 . A method for treating or ameliorating one or more symptoms of a liver disease or liver disorder, the method comprising administering the rAAV particle of claim 4 to a human subject diagnosed with the liver disease or liver disorder.

18 . The method of claim 17 , wherein the liver disease or liver disorder is liver cancer.

19 . The method of claim 18 , wherein the liver cancer is human hepatocellular carcinoma (HCC).

20 . A method of transducing a population of liver cells or liver tumor cells in a human diagnosed with, having, or suspected of having hepatocellular carcinoma (HCC); the method comprising administering to the human, a composition that comprises an effective amount of the rAAV particle of claim 4 , for a time effective to transduce the population of liver cells or liver tumor cells.

Assignments (2)

Continuity (4)

References Cited (256)