Compositions and methods for treatment of cancer using bacteria
Provided herein are compositions comprising substantially non-viable Gram-negative bacterial organisms that have a substantial reduction in endotoxin activity and/or pyrogenicity and methods for treating a cancer using the same. Also provided are methods for treating cancer provided herein, comprising administering to a mammal diagnosed with cancer, substantially non-viable Gram-negative bacteria having a substantial reduction in endotoxin activity and/or pyrogenicity, in an amount sufficient to inhibit growth or metastasis of the cancer. An additional method is provided comprising administering viable or non-viable Gram-negative bacterial organisms that have a genetic defect that results in a substantial loss of lipopolysaccharide within the outer membrane of the bacteria. Further provided are methods for reducing endotoxin activity and/or pyrogenicity in Gram-negative bacteria comprising treatment with polymyxin and glutaraldehyde.
1. A pharmaceutical composition comprising substantially non-viable and intact Gram-negative bacterial cells having at least 80% reduction of lipopolysaccharide (LPS)-mediated endotoxin activity as compared to the corresponding wild-type Gram-negative bacterial cells, wherein the substantially non-viable and intact Gram-negative bacterial cells are prepared by treating live Gram-negative bacterial cells with 200 to 5000 micrograms of a polymyxin per 1×10 7 to 5×10 10 of the live Gram-negative bacterial cells per mL at a temperature of 2° C. to 10° C. and with glutaraldehyde at a concentration of from 0.5% to 2.0%, wherein the polymyxin is selected from the group consisting of polymyxin B or polymyxin E.
2. The pharmaceutical composition of claim 1 , wherein the substantially non-viable and intact Gram-negative bacterial cells have at least 90% reduction of the LPS-mediated endotoxin activity as compared to the corresponding wild-type Gram-negative bacterial cells.
3. The pharmaceutical composition of claim 1 , wherein the reduction of the LPS-mediated endotoxin activity is as measured by an in vitro Limulus amebocyte lysate (LAL) assay.
4. The pharmaceutical composition of claim 1 , wherein the Gram-negative bacterial cells are Salmonella cells or Escherichia cells.
5. The pharmaceutical composition of claim 1 , wherein the polymyxin is polymyxin B.
6. The pharmaceutical composition of claim 1 , wherein the treatment with the polymyxin is at a temperature of 4° C.
7. The pharmaceutical composition of claim 5 , wherein the polymyxin B is present at a concentration of from 1000 micrograms/mL to 3000 micrograms/m L.
8. The pharmaceutical composition of claim 1 , wherein the glutaraldehyde is present at a concentration of from 0.5% to 1.0%.
9. The pharmaceutical composition of claim 5 , wherein the polymyxin B is present with magnesium in the form of magnesium chloride.
10. A method of treating a cancer in a mammalian cancer patient comprising administering to the patient an effective amount of the pharmaceutical composition of claim 1 .