Compositions and methods of delivering treatments for latent viral infections

• US 10,066,241 B2
• Filed: 05/29/2015
• Issued: 09/04/2018
• Est. Priority Date: 05/30/2014
• Status: Expired due to Fees

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Inventors: Quake, Stephen R., Wang, Jianbin
• Primary Examiner(s): Leavitt, Maria G
• Application Number:
  US14/725,943
• Publication Number:
  US 20150368670A1
• Time in Patent Office: 1,194 Days
• Field of Search: None
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/1133 against herpetoviridae, e.g. HSV
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 2310/20 involving clustered regularly interspaced short palindromic repeats [CRISPRs]
• CPC Class Codes: C12N 2330/51 Specially adapted vectors
• CPC Class Codes: C12N 2810/60 from viruses
• CPC Class Codes: C12N 2820/60 from viruses
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12Y 301/00 Hydrolases acting on ester bonds (3.1)
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides delivery methods and compositions for antiviral therapeutics. Methods and compositions are provided for targeted delivery of antiviral therapeutics into cells of interest using, for example, viral vectors such as adenovirus, AAV, and replication incompetent HSV. These and other delivery systems can be used as vehicles to deliver DNA vectors encoding a nuclease or a cell-killing gene. These delivery methods can also be used to deliver naked DNA or RNA, protein products, plasmids containing a promoter that is active only in a latent viral state which drives a cell-killing gene, or other therapeutic agents.

Claims

1. A composition for treating an Epstein-Barr virus (EBV) viral infection, the composition comprising:

a CRISPR/Cas9 plasmid comprising (a) a nucleic acid molecule under the control of a regulatory sequence and encoding a guide RNA that hybridizes to a complementary sequence in a target EBV genome, wherein the complementary sequence in the target EBV genome comprises a sequence selected from the group consisting of SEQ ID NO;

1, SEQ ID NO;

2, SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, and SEQ ID NO;

7;

(b) a nucleic acid molecule under the control of a regulatory sequence and encoding a gene for a Cas9 endonuclease; and

(c) an EBV origin of replication;

wherein delivery of the plasmid to EBV-infected eukaryotic cells causes the EBV-infected eukaryotic cells to express the guide RNA and the Cas9 endonuclease leading to reduced EBV load relative to untreated EBV-infected eukaryotic cells.

2. The composition of claim 1, wherein the plasmid encodes a plurality of guide RNAs.

3. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

2.

4. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

6.

5. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

1 and further wherein the plasmid encodes;

a second guide RNA that hybridizes to a second complementary sequence in the target EBV genome comprising SEQ ID NO;

2; and

a third guide RNA that hybridizes to a third complementary sequence in the target EBV genome comprising SEQ ID NO;

6.

6. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

4 and further wherein the plasmid encodes a second guide RNA that hybridizes to a second complementary sequence in the target EBV genome comprising SEQ ID NO;

5.

7. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

3.

8. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

4 and further wherein the plasmid encodes;

a second guide RNA that hybridizes to a second complementary sequence in the target EBV genome comprising SEQ ID NO;

5; and

a third guide RNA that hybridizes to a third complementary sequence in the target EBV genome comprising SEQ ID NO;

3.

9. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

1 and further wherein the plasmid encodes;

a second guide RNA that hybridizes to a second complementary sequence in the target EBV genome comprising SEQ ID NO;

2;

a third guide RNA that hybridizes to a third complementary sequence in the target EBV genome comprising SEQ ID NO;

3;

a fourth guide RNA that hybridizes to a fourth complementary sequence in the target EBV genome comprising SEQ ID NO;

4;

a fifth guide RNA that hybridizes to a fifth complementary sequence in the target EBV genome comprising SEQ ID NO;

5;

a sixth guide RNA that hybridizes to a sixth complementary sequence in the target EBV genome comprising SEQ ID NO;

6; and

a seventh guide RNA that hybridizes to a seventh complementary sequence in the target EBV genome comprising SEQ ID NO;

7.

10. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

3 and further wherein the plasmid encodes a second guide RNA that hybridizes to a second complementary sequence in the target EBV genome comprising SEQ ID NO;

7.

11. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

1.

12. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

5.

13. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

7.

14. The composition of claim 1, wherein the complementary sequence in the target EBV genome comprises SEQ ID NO:

4.