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 and administering electroporation therapy to the tumor; and b) administering an effective dose of an immune checkpoint inhibitor to the subject concurrently with step a).
2. The method of claim 1 , wherein the electroporation therapy comprises 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 the at least one voltage pulses has a field strength of about 20 V/cm to about 1500 V/cm.
4. The method of claim 3 , wherein the electroporation therapy comprises administration of 1-6 voltage pulses.
5. The method of claim 1 , wherein the immune checkpoint inhibitor is a PD-1 or PD-L1 antagonist.
6. The method of claim 5 , 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.
7. The method of claim 5 , wherein the PD-1 or PD-L1 antagonist is administered systemically.
8. The method of claim 5 , wherein the PD-1 or PD-L1 antagonist is an anti-PD-1 or anti-PD-L1 antibody.
9. The method of claim 8 , wherein the PD-1 or PD-L1 antagonist is selected from the group consisting of: nivolumab, pembrolizumab, pidilizumab, and MPDL3280A.
10. The method of claim 1 , wherein the at least one plasmid coding for the at least one immunostimulatory cytokine comprises a nucleic acid encoding an IL-12.
11. The method of claim 10 , wherein the at least one plasmid coding for the at least one immunostimulatory cytokine comprises a nucleic acid encoding an IL-12 p35 subunit and an IL-12 p40 subunit.
12. The method of claim 11 , wherein the 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.
13. The method of claim 1 , wherein the treatment-refractory cutaneous or subcutaneous cancerous tumor is selected from the group consisting of: breast cancer tumor triple-negative breast cancer tumor, and melanoma tumor.
14. The method of claim 1 , wherein the method further comprises injecting the tumor with a second effective dose of the at least one plasmid coding for the at least one immunostimulatory cytokine and administering electroporation therapy to the tumor at a time 4 days after step a) and injecting the tumor with a third effective dose of the at least one plasmid coding for the at least one immunostimulatory cytokine and administering electroporation therapy to the tumor at a time 7 days after step a).