Tripartite cancer theranostic nucleic acid constructs

Patent metadata

Field	Value
Publication number	US-10166300-B2
Application number	US-201414895556-A
Country	US
Kind code	B2
Filing date	Jun 4, 2014
Priority date	Jun 4, 2013
Publication date	Jan 1, 2019
Grant date	Jan 1, 2019

Abstract

The presently disclosed subject matter generally relates to genetic constructs and methods for their use in cancer imaging, cancer treatment, and combined imaging and treatment protocols. In particular, the presently disclosed subject matter relates to tripartite cancer theranostic nucleic acid constructs that permit simultaneous cancer specific viral replication, expression of a diagnostic gene product, and expression of a therapeutic gene.

First claim

That which is claimed:

1. A composition comprising a population of ultrasound targeted microbubbles, wherein the microbubbles comprise a lipid layer that stably binds a tripartite nucleic acid construct, and wherein the tripartite nucleic acid construct comprises a first promoter operably linked to at least one gene required for viral replication, a second promoter operably linked to a reporter gene , and a third promoter operably linked to a therapeutic gene, wherein the first promoter and the second promoter are each cancer-selective, and wherein the second promoter is SEQ. ID. NO. 15.

2. The composition of claim 1 , wherein the nucleic acid construct is selected from the group consisting of adenoviral, lentiviral, retroviral, adeno-associated viral, and herpes simplex viral.

3. The composition of claim 2 , wherein the nucleic acid construct is an adenoviral vector.

4. The composition of claim 1 , wherein the first promoter is a progression-elevated gene-3 promoter (PEG-Prom).

5. The composition of claim 1 , wherein the at least one gene required for viral replication is adenovirus early region IA (E1A) or early region 1B (E1B).

6. The composition of claim 1 , further comprising an imaging agent selected from the group consisting of (β-galactosidase, luciferase, horse radish peroxidase, thymidine kinase, and alkaline phosphatase.

7. The composition of claim 6 , wherein the imaging agent is herpes simplex virus thymidine kinase (HSV-tk).

8. The composition of claim 1 , wherein the third promoter is a constitutive promoter.

9. The composition of claim 8 , wherein the third promoter is cytomegalovirus (CMV) promoter.

10. The composition of claim 1 , wherein the third promoter is a cancer-selective promoter.

11. The composition of claim 1 , wherein the therapeutic agent is selected from the group consisting of an anti-cancer agent, an apoptosis inducing agent, a tumor suppressor agent, an immune system enhancing agent, and an immunomodulatory agent.

12. The composition of claim 1 , wherein the therapeutic agent is an immunomodulatory cytokine.

13. The composition of claim 12 , wherein the immunomodulatory cytokine is mda-7/IL-24.

14. The composition of claim 12 , wherein the immunomodulatory cytokine is M4.

15. A composition comprising a population of ultrasound targeted microbubbles, wherein the microbubbles comprise a lipid layer that stably binds a tripartite nucleic acid construct, and wherein the tripartite nucleic acid construct comprises a first promoter operably linked to at least one gene required for viral replication, a second promoter operably linked to a reporter gene, and a third promoter operably linked to a therapeutic gene, wherein:
(a) the first promoter is a progression-elevated gene-3 promoter (PEG-Prom);
(b) the at least one gene required for viral replication is adenovirus early region IA (E1A) or early region 1B (E1B);
(c) the second promoter is SEQ. ID. NO. 15;
(d) the third promoter is cytomegalovirus (CMV) promoter; and
(e) the therapeutic gene is an immunomodulatory cytokine selected from the group consisting of mda-7/IL-24 and M4.

16. The composition of claim 15 , further comprising an imaging agent selected from the group consisting of β-galactosidase, luciferase, horse radish peroxidase, thymidine kinase, and alkaline phosphatase.

17. The composition of claim 16 , wherein the imaging agent is herpes simplex virus thymidine kinase (HSV-tk).

Assignees

• Univ Virginia Commonwealth
• Univ Johns Hopkins

Inventors

• Fisher Paul B
• Das Swadesh K
• Menezes Mitchell E
• Sarkar Devanand
• Pomper Martin G

Classifications

• A61K49/0045

  the fluorescent agent being a peptide or protein used for imaging or diagnosis in vivo · CPC title

• A61K41/0028

  Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds · CPC title

• A61K48/0058 Primary

  Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct · CPC title

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title

• C12N2710/10343

  viral genome or elements thereof as genetic vector · CPC title