POLYMERIC NANOPARTICLES PROVIDING NUCLEIC ACIDS ENCODING TNF-a
The present invention relates to polymeric nanoparticles comprising a cytokine or a nucleic acid encoding for a cytokine, pharma-ceutical compositions comprising the same, and methods for treating certain diseases comprising administering these polymeric nanoparticles to a subject in need thereof.
1 - 35 . (canceled)
36 . A method for inducing TNF expression in a cell comprising contacting the cell with a composition comprising:
a) a polymeric nanoparticle comprising a block copolymer comprising poly(lactic acid) (PLA) and poly(ethylene glycol) (PEG); and
b) an isolated nucleic acid that encodes a protein comprising an amino acid sequence at least 70% identical to the sequence of SEQ ID NO:9, or a portion thereof.
37 . The method of claim 36 , wherein the polymeric nanoparticle comprises a poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) di-block copolymer.
38 . The method of claim 36 , wherein the polymeric nanoparticle comprises a poly(lactic acid)-poly(ethylene glycol)-poly(propylene glycol)-poly(ethylene glycol)-poly(lactic acid) (PLA-PEG-PPG-PEG-PLA) block copolymer.
39 . The method of claim 36 , wherein the molecular weight of PLA is between about 10,000 and about 100,000 daltons.
40 . The method of claim 36 , wherein the composition further comprises a chemotherapeutic agent or a targeted anti-cancer agent selected from the group consisting of doxorubicin, daunorubicin, decitabine, irinotecan, SN-38, cytarabine, docetaxel, triptolide, geldanamycin, 17-AAG, 5-FU, oxaliplatin, carboplatin, methotrexate, paclitaxel, indenoisoquinolines, and bortezomib.
41 . The method of claim 36 , wherein the polymeric nanoparticles further comprise a targeting moiety attached to the outside of the polymeric nanoparticles, and wherein the targeting moiety is an antibody, peptide, or aptamer.
42 . The method of claim 36 , wherein the isolated nucleic acid comprises a nucleotide sequence at least 95% identical to nucleotides 404-874 of SEQ ID NO:11.
43 . The method of claim 36 , wherein the isolated nucleic acid comprises nucleotides 404-874 of SEQ ID NO:11.
44 . The method of claim 36 , wherein the isolated nucleic acid comprises a nucleotide sequence at least 95% identical to nucleotides 176-874 of SEQ ID NO:11.
45 . The method of claim 36 , wherein the isolated nucleic acid comprises nucleotides 176-874 of SEQ ID NO:11.
46 . The method of claim 36 , wherein the isolated nucleic acid comprises a nucleotide sequence at least 95% identical to the nucleotide sequence of SEQ ID NO:11.
47 . The method of claim 36 , wherein the isolated nucleic acid comprises the nucleotide sequence of SEQ ID NO:11.
48 . The method of claim 36 , wherein the isolated nucleic acid encodes an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:9.
49 . The method of claim 36 , wherein the isolated nucleic acid encodes the amino acid sequence of SEQ ID NO:9.
50 . The method of claim 36 , wherein the isolated nucleic acid encodes an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:10.
51 . The method of claim 36 , wherein the isolated nucleic acid encodes the amino acid sequence of SEQ ID NO:10.
52 . The method of claim 36 , wherein the composition further comprises a cationic peptide.
53 . The method of claim 36 , wherein the composition further comprises a cell-penetrating peptide.
54 . The method of claim 36 , wherein the composition further comprises a cationic cell-penetrating peptide.
55 . The method of claim 54 , wherein the isolated nucleic acid and the cationic cell-penetrating peptide form a complex.
56 . The method of claim 54 , wherein the cationic cell-penetrating peptide is selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, and polyarginine.
57 . A method for treating cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising:
a) a polymeric nanoparticle comprising a block copolymer comprising poly(lactic acid) (PLA) and poly(ethylene glycol) (PEG); and
b) an isolated nucleic acid that encodes a protein comprising an amino acid sequence at least 70% identical to the sequence of SEQ ID NO:9, or a portion thereof.
58 . The method of claim 57 , wherein the polymeric nanoparticle comprises a PLA-PEG di-block copolymer.
59 . The method of claim 57 , wherein the polymeric nanoparticle comprises a PLA-PEG-PPG-PEG-PLA block copolymer.
60 . The method of claim 57 , wherein the molecular weight of PLA is between about 10,000 and about 100,000 daltons.
61 . The method of claim 57 , wherein the pharmaceutical composition further comprises a chemotherapeutic agent or a targeted anti-cancer agent selected from the group consisting of doxorubicin, daunorubicin, decitabine, irinotecan, SN-38, cytarabine, docetaxel, triptolide, geldanamycin, 17-AAG, 5-FU, oxaliplatin, carboplatin, methotrexate, paclitaxel, indenoisoquinolines, and bortezomib.
62 . The method of claim 57 , wherein the polymeric nanoparticles further comprise a targeting moiety attached to the outside of the polymeric nanoparticles, and wherein the targeting moiety is an antibody, peptide, or aptamer.
63 . The method of claim 57 , wherein the cancer is breast cancer, prostate cancer, non-small cell lung cancer, metastatic colon cancer, pancreatic cancer, or a hematological malignancy.
64 . The method of claim 57 , wherein the cancer comprises a PD-1 refractory tumor.
65 . The method of claim 57 , wherein the isolated nucleic acid comprises a nucleotide sequence at least 95% identical to nucleotides 404-874 of SEQ ID NO:11.
66 . The method of claim 57 , wherein the isolated nucleic acid comprises nucleotides 404-874 of SEQ ID NO:11.
67 . The method of claim 57 , wherein the isolated nucleic acid comprises a nucleotide sequence at least 95% identical to nucleotides 176-874 of SEQ ID NO:11.
68 . The method of claim 57 , wherein the isolated nucleic acid comprises nucleotides 176-874 of SEQ ID NO:11.
69 . The method of claim 57 , wherein the isolated nucleic acid comprises a nucleotide sequence at least 95% identical to the nucleotide sequence of SEQ ID NO:11.
70 . The method of claim 57 , wherein the isolated nucleic acid comprises the nucleotide sequence of SEQ ID NO:11.
71 . The method of claim 57 , wherein the isolated nucleic acid encodes an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:9.
72 . The method of claim 57 , wherein the isolated nucleic acid encodes the amino acid sequence of SEQ ID NO:9.
73 . The method of claim 57 , wherein the isolated nucleic acid encodes an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO:10.
74 . The method of claim 57 , wherein the isolated nucleic acid encodes the amino acid sequence of SEQ ID NO:10.
75 . The method of claim 57 , wherein the pharmaceutical composition further comprises a cationic peptide.
76 . The method of claim 57 , wherein the pharmaceutical composition further comprises a cell-penetrating peptide.
77 . The method of claim 57 , wherein the pharmaceutical composition further comprises a cationic cell-penetrating peptide.
78 . The method of claim 77 , wherein the isolated nucleic acid and the cationic cell-penetrating peptide form a complex.
79 . The method of claim 77 , wherein the cationic cell-penetrating peptide is selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, and polyarginine.