IP Library › Granted Patent US 10,137,144
Granted Patent B2
US 10,137,144 · App. 15/904,131 · Granted Nov 27, 2018

Methods and compositions for the activation of gamma-delta T-cells

Inventors: Charles David Pauza (Baltimore, MD); Haishan Li (North Potomac, MD); Tyler Lahusen (Frederick, MD); Mei-Ling Liou (Germantown, MD)
Assignee: American Gene Technologies International Inc.
A61K31/7105A61K31/675A61K47/6807A61K47/6901A61P35/02A61K2300/00
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Quick Facts
Patent No.
US 10,137,144
App. No.
15/904,131
Granted
Nov 27, 2018
Kind
B2
Abstract

The present invention relates generally to methods and compositions for gene therapy and immunotherapy that activate gamma delta T-cells, and in particular, can be used in the treatment of various cancers and infectious diseases.

Claims (329)

1. A method of treating a cancer in a subject using an immunotherapy-based composition, the method comprising:

administering a therapeutically-effective amount of an bisphosphonate drug to the subject; and

administering a therapeutically-effective amount of the immunotherapy-based composition to the subject,

wherein the immunotherapy-based composition comprises a lentiviral particle, the lentiviral particle comprising:

a. an envelope protein capable of infecting one or more cancer cells, and

b. at least one encoded shRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded shRNA comprises a sequence having at least 80% percent identity with:

i.

(SEQ ID NO: 1)

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

ii.

(SEQ ID NO: 2)

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

iii.

(SEQ ID NO: 3)

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

iv.

(SEQ ID NO: 4)

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCT

TCTGCTTTTT;

or

c. at least one encoded microRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded microRNA comprises a sequence having at least 80% percent identity with:

i.

(SEQ ID NO: 5)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTG

CTGCCTACTGCCTCGGACTTCAAGGGGCT;

ii.

(SEQ ID NO: 6)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCT

GCCTACTGCCTCGGACTTCAAGGGGCT;

iii.

(SEQ ID NO: 7)

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAA

GCCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCT

CGGA;

iv.

(SEQ ID NO: 8)

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCC

TTCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTC

AGGACACAAGGCCTGTTACTAGCACTCA;

v.

(SEQ ID NO: 9)

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCC

TTCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCT

GACATTTTGGTATCTTTCATCTGACCA;

or

vi.

(SEQ ID NO: 10)

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTC

CTTCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCT

TCCCTCCCAATGACCGCGTCTTCGTCG.

2. The method of claim 1 , wherein the at least one encoded shRNA comprises a sequence having at least 85% percent identity with:

i.

(SEQ ID NO: 1)

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

ii.

(SEQ ID NO: 2)

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

iii.

(SEQ ID NO: 3)

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

iv.

(SEQ ID NO: 4)

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCT

TCTGCTTTTT;

or

wherein the at least one encoded microRNA comprises a sequence having at least 85% percent identity with:

v.

(SEQ ID NO: 5)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTG

CTGCCTACTGCCTCGGACTTCAAGGGGCT;

vi.

(SEQ ID NO: 6)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCT

GCCTACTGCCTCGGACTTCAAGGGGCT;

vii.

(SEQ ID NO: 7)

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAA

GCCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCT

CGGA;

viii.

(SEQ ID NO: 8)

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCC

TTCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTC

AGGACACAAGGCCTGTTACTAGCACTCA;

ix.

(SEQ ID NO: 9)

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCC

TTCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCT

GACATTTTGGTATCTTTCATCTGACCA;

or

x.

(SEQ ID NO: 10)

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTC

CTTCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCT

TCCCTCCCAATGACCGCGTCTTCGTCG.

3. The method of claim 1 , wherein the at least one encoded shRNA comprises a sequence having at least 90% percent identity with:

i.

(SEQ ID NO: 1)

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

ii.

(SEQ ID NO: 2)

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

iii.

(SEQ ID NO: 3)

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

iv.

(SEQ ID NO: 4)

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCT

TCTGCTTTTT;

or

wherein the at least one encoded microRNA comprises a sequence having at least 90% percent identity with:

v.

(SEQ ID NO: 5)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTG

CTGCCTACTGCCTCGGACTTCAAGGGGCT;

vi.

(SEQ ID NO: 6)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCT

GCCTACTGCCTCGGACTTCAAGGGGCT;

vii.

(SEQ ID NO: 7)

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAA

GCCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCT

CGGA;

viii.

(SEQ ID NO: 8)

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCC

TTCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTC

AGGACACAAGGCCTGTTACTAGCACTCA;

ix.

(SEQ ID NO: 9)

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCC

TTCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCT

GACATTTTGGTATCTTTCATCTGACCA;

or

x.

(SEQ ID NO: 10)

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTC

CTTCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCT

TCCCTCCCAATGACCGCGTCTTCGTCG.

4. The method of claim 1 , wherein the at least one encoded shRNA comprises a sequence having at least 95% percent identity with:

i.

(SEQ ID NO: 1)

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

ii.

(SEQ ID NO: 2)

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

iii.

(SEQ ID NO: 3)

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

iv.

(SEQ ID NO: 4)

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCT

TCTGCTTTTT;

or

wherein the at least one encoded microRNA comprises a sequence having at least 95% percent identity with:

v.

(SEQ ID NO: 5)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTG

CTGCCTACTGCCTCGGACTTCAAGGGGCT;

vi.

(SEQ ID NO: 6)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCT

GCCTACTGCCTCGGACTTCAAGGGGCT;

vii.

(SEQ ID NO: 7)

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAA

GCCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCT

CGGA;

viii.

(SEQ ID NO: 8)

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCC

TTCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTC

AGGACACAAGGCCTGTTACTAGCACTCA;

ix.

(SEQ ID NO: 9)

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCC

TTCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCT

GACATTTTGGTATCTTTCATCTGACCA;

or

x.

(SEQ ID NO: 10)

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTC

CTTCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCT

TCCCTCCCAATGACCGCGTCTTCGTCG.

5. The method of claim 1 , wherein the at least one encoded shRNA comprises:

i.

(SEQ ID NO: 1)

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

ii.

(SEQ ID NO: 2)

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

iii.

(SEQ ID NO: 3)

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

iv.

(SEQ ID NO: 4)

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCT

TCTGCTTTTT;

or

wherein the at least one encoded microRNA comprises:

v.

(SEQ ID NO: 5)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTG

CTGCCTACTGCCTCGGACTTCAAGGGGCT;

vi.

(SEQ ID NO: 6)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCT

GCCTACTGCCTCGGACTTCAAGGGGCT;

vii.

(SEQ ID NO: 7)

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAA

GCCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCT

CGGA;

viii.

(SEQ ID NO: 8)

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCC

TTCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTC

AGGACACAAGGCCTGTTACTAGCACTCA;

ix.

(SEQ ID NO: 9)

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCC

TTCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCT

GACATTTTGGTATCTTTCATCTGACCA;

or

x.

(SEQ ID NO: 10)

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTC

CTTCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCT

TCCCTCCCAATGACCGCGTCTTCGTCG.

6. The method of claim 1 , wherein the one or more cancer cells are present in a cancer selected from one or more of a carcinoma, a leukemia, a lymphoma, a sarcoma, a myeloma, a mesothelioma, a mixed type, or mixtures thereof.

7. The method of claim 1 , wherein the bisphosphonate drug comprises zoledronic acid.

8. The method of claim 1 , wherein the bisphosphonate drug and the immunotherapy-based composition are administered in a fixed combination.

9. The method of claim 1 , wherein the bisphosphonate drug and the immunotherapy-based composition are administered in a non-fixed combination.

10. The method of claim 9 , wherein the bisphosphonate drug and the immunotherapy-based composition are administered simultaneously.

11. The method of claim 9 , wherein the bisphosphonate drug and the immunotherapy-based composition are administered sequentially.

12. The method of claim 1 , wherein the bisphosphonate drug and the immunotherapy-based composition are administered in synergistically effective amounts.

13. The method of claim 1 , wherein the bisphosphonate drug and the immunotherapy-based composition are administered at a synergistically effective time interval.

14. The method of claim 1 , wherein the one or more cancer cells are capable of activating a gamma delta T cell resident in the subject following infection of the one or more cancer cells with the immunotherapy-based composition.

15. The method of claim 14 , wherein activating the gamma delta T cell comprises increasing tumor necrosis factor (TNF)-α expression by the gamma delta T cell.

16. The method of claim 1 , wherein the enzyme of the mevalonate pathway is farnesyl diphosphate synthase (FDPS).

17. A pharmaceutical combination comprising:

a bisphosphonate compound; and

a lentiviral particle produced by a packaging cell and capable of infecting a target cell, the lentiviral particle comprising an envelope protein capable of infecting the target cell, and:

a. at least one encoded shRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded shRNA comprises a sequence having at least 80% percent identity with:

i.

(SEQ ID NO: 1)

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

ii.

(SEQ ID NO: 2)

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

iii.

(SEQ ID NO: 3)

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

iv.

(SEQ ID NO: 4)

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCT

TCTGCTTTTT;

or

b. at least one encoded microRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded microRNA comprises a sequence having at least 80% percent identity with:

v.

(SEQ ID NO: 5)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTG

CTGCCTACTGCCTCGGACTTCAAGGGGCT;

vi.

(SEQ ID NO: 6)

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCT

GCCTACTGCCTCGGACTTCAAGGGGCT;

vii.

(SEQ ID NO: 7)

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAA

GCCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCT

CGGA;

viii.

(SEQ ID NO: 8)

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCC

TTCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTC

AGGACACAAGGCCTGTTACTAGCACTCA;

ix.

(SEQ ID NO: 9)

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCC

TTCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCT

GACATTTTGGTATCTTTCATCTGACCA;

or

x.

(SEQ ID NO: 10)

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTC

CTTCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCT

TCCCTCCCAATGACCGCGTCTTCGTCG;

wherein the pharmaceutical combination is at least one of fixed and non-fixed.

18. The pharmaceutical combination of claim 17 , wherein the pharmaceutical composition comprises a fixed combination.

19. The pharmaceutical combination of claim 17 , wherein the pharmaceutical composition comprises a non-fixed combination.

20. The pharmaceutical combination of claim 1 , wherein the bisphosphonate drug comprises zoledronic acid.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 23, 2018
From: PAUZA, CHARLES DAVID; LI, HAISHAN; LAHUSEN, TYLER; LIOU, MEI-LING
To: AMERICAN GENE TECHNOLOGIES INTERNATIONAL INC.
Reel/Frame 047281/0064 →
Continuity (4)
Continuation In Part 15652080 · Jul 17, 2017
Continuation PCTUS2017013399 · Jan 13, 2017
Provisional Application 62279474 · Jan 15, 2016
Related Publication 20180256624A1 · Sep 13, 2018
Cited By (8)
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