Method for the treatment of malignancies
The present invention provides for the intratumoral delivery of at least one immunostimulatory cytokine in combination with at least one checkpoint inhibitor. In particular, it provides delivery of a plasmid encoding the immunostimulatory cytokine using intratumoral electroporation. The checkpoint inhibitor may be administered systemically or encoded on a plasmid and delivered using intratumoral electroporation. The checkpoint inhibitor may be delivered contemporaneously with or after treatment with the immunomodulatory cytokine.
1. A method of treating a subject having a treatment-refractory, cutaneous or subcutaneous, cancerous tumor, the method comprising:
a) injecting the treatment-refractory cutaneous or subcutaneous cancerous tumor with an effective dose of at least one plasmid coding for at least one immunostimulatory cytokine;
b) administering electroporation therapy to the tumor; and
c) administering an effective dose of an immune checkpoint inhibitor to the subject.
2. The method of claim 1 , wherein the electroporation therapy comprises the administration of at least one voltage pulse over a duration of about 100 microseconds to about 1 millisecond.
3. The method of claim 2 , wherein at least one voltage pulse delivered to the tumor has a field strength of about 200 V/cm to about 1500 V/cm.
4. The method of claim 1 , wherein the immune checkpoint inhibitor is a PD-1 or PD-L1 antagonist.
5. The method of claim 4 , wherein the PD-1 or PD-L1 antagonist is encoded on the plasmid encoding the immunostimulatory cytokine or on a second plasmid and delivered to the cancerous tumor by electroporation therapy.
6. The method of claim 4 , wherein the PD-1 or PD-L1 antagonist is administered systemically.
7. The method of claim 6 , wherein the PD-1 or PD-L1 antagonist is selected from the group consisting of: nivolumab, pembrolizumab, pidilizumab, and MPDL3280A.
8. The method of claim 4 , wherein the PD-1 or PD-L1 antagonist is administered after electroporation of the tumor.
9. The method of claim 1 , wherein the plasmid encoding for the at least one immunostimulatory cytokine comprises a nucleic acid encoding an IL-12 p35 subunit and an IL-12 p40 subunit.
10. The method of claim 1 , wherein the treatment-refractory cutaneous or subcutaneous cancerous tumor is a treatment-refractory, cutaneous or subcutaneous, breast cancer tumor or a treatment-refractory, cutaneous or subcutaneous, triple-negative breast cancer tumor.
11. The method of claim 1 , wherein the cancerous tumor is melanoma.
12. A method of treating a subject having a treatment-refractory, cutaneous or subcutaneous, cancerous tumor comprising:
a) administering a first treatment at a first time (T1), wherein the first treatment comprises injecting the cancerous tumor with a first effective dose of at least one plasmid encoding at least one immunostimulatory cytokine and administering a first electroporation therapy to the cancerous tumor, wherein the first electroporation therapy further comprises administration of at least one voltage pulse having a duration of about 100 microseconds to about 1 millisecond;
b) administering a second treatment at a second time (T2) after T1, wherein the second treatment comprises injecting the cancerous tumor with a second effective dose of at least one plasmid encoding at least one immunostimulatory cytokine and administering a second electroporation therapy to the cancerous tumor, wherein the second electroporation therapy further comprises administration of at least one voltage pulse having a duration of about 100 microseconds to about 1 millisecond;
c) administering a third treatment at a third time (T3) after T2, wherein the third treatment comprises injecting the cancerous tumor with a third effective dose of at least one plasmid encoding at least one immunostimulatory cytokine and administering a third electroporation therapy to the cancerous tumor, wherein the third electroporation therapy further comprises administration of at least one voltage pulse having a duration of about 100 microseconds to about 1 millisecond; and
d) administering a fourth treatment at a fourth time (T4) after T3, wherein the fourth treatment comprises administering an effective dose of an immune checkpoint inhibitor to the subject.
13. The method of claim 12 , wherein:
a) T1 is day 1;
b) T2 is day 5;
c) T3 is day 8; and
d) T4 is day 20-120.
14. The method of claim 12 , wherein the method further comprises:
a) at least one cycle of intratumoral delivery of the at least one plasmid encoding for the at least one immunostimulatory cytokine by electroporation;
b) an interval of at least 20-120 days following the one cycle; and
c) administration to the subject, the immune checkpoint inhibitor or a combination of at least one immune checkpoint inhibitor with at least one additional cycle of intratumoral electroporation therapy.
15. The method of claim 1 , wherein said at least one plasmid coding for the at least one immunostimulatory cytokine is a plasmid encoding an IL-12 p35 subunit and an IL-12 p40 subunit separated by an internal ribosomal entry site.
16. The method of claim 4 , wherein the PD-1 or PD-L1 antagonist is an anti-PD-1 or anti-PD-L1 antibody.
17. The method of claim 1 , wherein said at least one plasmid coding for the at least one immunostimulatory cytokine is a plasmid encoding an IL-12.
18. The method of claim 12 , wherein the treatment-refractory, cutaneous or subcutaneous, cancerous tumor is a treatment-refractory, cutaneous or subcutaneous, melanoma tumor or lesion.