Compositions comprising bacterially derived minicells and methods of using the same

1. A composition comprising:

(a) a therapeutically effective dose of purified, intact bacterially derived minicells comprising at least one anti-neoplastic agent, wherein the anti-neoplastic agent comprises nemorubicin, PNU-159682, idarubicin, daunorubicin, caminomycin, and/or and

(b) an interferon type I agonist, an interferon type II agonist, or a combination of an interferon type I agonist and an interferon type II agonist,

wherein the interferon type I agonist is an oligonucleotide, wherein the oligonucleotide is double stranded DNA or DNA-RNA hybrids and comprises a sequence of at least about 40 nucleotides, and

wherein the interferon type II agonist is selected from the group consisting of C-glycosidific form of α-galactosylceramide (α-C-GalCer), α-galactosylceramide (α-GalCer), 12 carbon acyl form of galactosylceramide (β-GalCer), β-D-glucopyranosylceramide (β-GlcCer), 1,2-Diacyl-3-0-galactosyl-sn-glycerol (BbGL-II), diacylglycerol containing glycolipids (Glc-DAG-s2), ganglioside (GD3), gangliotriaosylceramide (Gg3Cer), glycosylphosphatidylinositol (GPI), α-glucuronosylceramide (GSL-1 or GSL-4), isoglobotrihexosylceramide (iGb3), lipophosphoglycan(LPG), lyosphosphatidylcholine (LPC), a-galactosylceramide analog (OCH), threitolceramide, and a combination thereof.

2. The composition of claim 1 , wherein element (b) of the composition comprises:

(i) a therapeutically effective dose of purified, intact bacterially derived minicells comprising an interferon type I agonist; or

(ii) a therapeutically effective dose of purified, intact bacterially derived minicells comprising an interferon type II agonist; or

(iii) a combination of:

(1) a therapeutically effective dose of purified, intact bacterially derived minicells comprising an interferon type I agonist; and

(2) a therapeutically effective dose of purified, intact bacterially derived minicells comprising an interferon type II agonist.

3. The composition of claim 1 , wherein:

(a) the anti-neoplastic agent and the interferon type I agonist, the interferon type II agonist, or the combination of an interferon type I agonist and an interferon type II agonist, are packaged within two or more purified, intact bacterially derived minicells; or

(b) the anti-neoplastic agent and the interferon type I agonist, the interferon type II agonist, or the combination of an interferon type I agonist and an interferon type II agonist are packaged within three separate populations of purified, intact bacterially derived minicells.

4. The composition of claim 1 comprising the anti-neoplastic agent, the interferon type I agonist, and the interferon type II agonist, wherein:

(a) the anti-neoplastic agent, the interferon type I agonist, and the interferon type II agonist are comprised within the same minicell;

(b) the anti-neoplastic agent and the interferon type I agonist are comprised within a first minicell, and the interferon type II agonist is comprised within a second minicell;

(c) the anti-neoplastic agent and the interferon type II agonist are comprised within a first minicell, and the interferon type I agonist is comprised within a second minicell;

(d) the anti-neoplastic agent is comprised within a first minicell, and the interferon type I agonist and the interferon type II agonist are comprised within a second minicell; or

(e) the anti-neoplastic agent is comprised within a first minicell, the interferon type I agonist is comprised within a second minicell, and the interferon type II agonist is comprised within a third minicell.

5. The composition of claim 1 , wherein the composition does not comprise an interferon type I agonist.

6. The composition of claim 1 , wherein:

the anti-neoplastic agent is PNU-159682.

7. The composition of claim 1 , wherein:

(a) the oligonucleotide comprises a sequence of at least about 50 nucleotides or at least about 60 nucleotides.

8. The composition of claim 1 , wherein the interferon type I agonist is an oligonucleotide selected from the group consisting of double stranded Z-DNA and B-DNA.

9. The composition of claim 1 , wherein: the interferon type II agonist is α-galactosylceramide (α-GalCer).

10. The composition of claim 1 , further comprising:

(a) a bispecific ligand bound to the minicells comprising the anti-neoplastic agent; and/or

(b) a bispecific ligand bound to the minicells comprising the type I interferon agonist; and/or

(c) a bispecific ligand bound to the minicells comprising the type II interferon agonist.

11. The composition according to claim 10 , wherein the bispecific ligand:

(a) comprises a first arm that carries specificity for a minicell surface structure and a second arm that carries specificity for a non-phagocytotic mammalian cell surface receptor; and/or

(b) comprises a first arm that carries specificity for a minicell surface structure and a second arm that carries specificity for a non-phagocytotic mammalian cell surface receptor and wherein the minicell surface structure is an O-polysaccharide component of a lipopolysaccharide on the minicell surface; and/or

(c) comprises a first arm that carries specificity for a minicell surface structure and a second arm that carries specificity for a non-phagocytotic mammalian cell surface receptor wherein the non-phagocytotic mammalian cell surface receptor is capable of activating receptor-mediated endocytosis of the minicell; and/or

(d) comprises a bispecific antibody or antibody fragment; and/or

(e) comprises a bispecific antibody or antibody fragment and wherein the antibody or antibody fragment comprises a first multivalent arm that carries specificity for a bacterially derived minicell surface structure and a second multivalent arm that carries specificity for a cancer cell surface receptor, wherein the cancer cell surface receptor is capable of activating receptor-mediated endocytosis of the minicell.

12. The composition of claim 1 , wherein the composition comprises fewer than about 1 contaminating parent bacterial cell per 10 7 minicells, fewer than about 1 contaminating parent bacterial cell per 10 8 minicells, fewer than about 1 contaminating parent bacterial cell per 10 9 minicells, fewer than about 1 contaminating parent bacterial cell per 10 10 minicells, or fewer than about 1 contaminating parent bacterial cell per 10 11 minicells.

13. The composition of claim 1 , further comprising a pharmaceutically acceptable carrier.

14. The composition of claim 1 , wherein the minicells are approximately 400 nm in diameter.

15. The composition of claim 1 , wherein the composition is free of parent bacterial cell contamination removable through 200 nm filtration.

16. The composition of claim 1 , wherein the composition comprises the following amount of minicells or killed bacterial cells:

(a) at least about 10 9 ;

(b) at least about 1×10 9 ;

(c) at least about 2×10 9 ;

(d) at least about 5×10 9 ;

(e) at least 8×10 9 ;

(f) no more than about 10 11 ;

(g) no more than about 1×10 11 ;

(h) no more than about 9×10 10 ; or

(i) no more than about 8×10 10 .

17. The composition of claim 1 , comprising the anti-neoplastic agent, the interferon type I agonist, and interferon gamma.