Use of Liposomes in a Carrier Comprising a Continuous Hydrophobic Phase as a Vehicle for Cancer Treatment

Abstract

The invention compositions comprising a continuous hydrophobic phase and liposomes as a vehicle for delivery of an antigen capable of inducing a cytotoxic T lymphocyte (CTL) response and methods for their use in the treatment of cancer.

1 . A composition comprising:
 a carrier comprising a continuous phase of a hydrophobic substance;   liposomes; and   at least one antigen capable of inducing a cytotoxic T lymphocyte (CTL) response.   
     
     
         2 . The composition of  claim 1  further comprising at least one T helper epitope. 
     
     
         3 . The composition of  claim 2 , wherein the T helper epitope is a separate molecule from the at least one antigen. 
     
     
         4 . The composition of  claim 2 , wherein the antigen comprises the T helper epitope. 
     
     
         5 . The composition of  claim 1 , wherein the antigen is a polypeptide. 
     
     
         6 . The composition of  claim 1 , wherein the antigen comprises a CTL epitope, or a combination of CTL epitopes. 
     
     
         7 . The composition of  claim 6 , wherein the CTL epitope is derived from a virus. 
     
     
         8 . The composition of  claim 7 , wherein CTL epitope is derived from human papilloma virus. 
     
     
         9 . The composition of  claim 8 , wherein the CTL epitope is an epitope of an E6 or E7 protein of human papilloma virus (HPV). 
     
     
         10 . The composition of  claim 9 , wherein the E6 epitope comprises the peptide sequence TIHDIILECV (T10V; SEQ ID NO: 5). 
     
     
         11 . The composition of  claim 9 , wherein the E7 epitope comprises a peptide sequence selected from the group consisting of: RAHYNIVTF (R9F; SEQ ID NO: 1), YMLDLQPETT Y10T; SEQ ID NO: 2), LLMGTLGIV (L9V; SEQ ID NO: 3), and TLGIVCPI (T81; SEQ ID NO: 4). 
     
     
         12 . The composition of  claim 9 , wherein the CTL epitope is a mixture of CTL epitopes of E6 and E7 proteins of HPV. 
     
     
         13 . The composition of  claim 12 , wherein the mixture of epitopes comprises Y10T (SEQ ID NO: 2), L9V (SEQ ID NO: 3), T81 (SEQ ID NO: 4) and T10V (SEQ ID NO: 5) peptide sequences. 
     
     
         14 . The composition of  claim 13 , wherein said epitopes are joined together to form one peptide (AB2) (SEQ ID NO: 14). 
     
     
         15 . The composition of  claim 6 , comprising a plurality of CTL epitopes linked to form a single polypeptide. 
     
     
         16 . The composition of  claim 6 , wherein the CTL epitope is derived from a tumor-associated protein. 
     
     
         17 . The composition of  claim 16  wherein the tumor associated protein is p53. 
     
     
         18 . The composition of  claim 16 , wherein the tumor-associated protein is tyrosinase-related protein-2 (TRP-2). 
     
     
         19 . The composition of  claim 6 , wherein the antigen comprises a mixture of a p53 epitope and a TRP-2 epitope. 
     
     
         20 . The composition of  claim 17 , wherein the CTL epitope is a p53 epitope comprising the peptide sequence KYICNSSCM (mK9M) (SEQ ID NO: 8). 
     
     
         21 . The composition of  claim 18 , wherein the CTL epitope is a TRP-2 epitope comprising the peptide sequence SVYDFFVWL (SEQ ID NO: 7). 
     
     
         22 . The composition of  claim 18 , wherein the CTL epitope is a TRP-2 epitope comprising the peptide sequence VYDFFVWL (SEQ ID NO: 6). 
     
     
         23 . The composition of  claim 1 , wherein the T helper epitope is a universal T helper epitope. 
     
     
         24 . The composition of  claim 23 , wherein the T helper epitope is PADRE (pan-DR epitope) (SEQ ID NO: 10). 
     
     
         25 . The composition of  claim 1 , wherein the T helper epitope is F21E (SEQ ID NO: 11). 
     
     
         26 . The composition of  claim 1 , wherein the T helper epitope is fused to the at least one antigen. 
     
     
         27 . The composition of  claim 26 , wherein the T helper epitope is PADRE and at least one antigen is a CTL epitope. 
     
     
         28 . The composition of  claim 1 , wherein the composition is capable of increasing IFN-γ producing cells. 
     
     
         29 . The composition of  claim 1 , further comprising an adjuvant. 
     
     
         30 . The composition of  claim 29 , wherein the adjuvant is a CpG oligodeoxynucleotide (CpG ODN) (SEQ ID NO: 12). 
     
     
         31 . The composition of  claim 29 , wherein the adjuvant is a lipopeptide. 
     
     
         32 . The composition of  claims 31 , wherein the lipopeptide is Pam3Cys-SKKKK. 
     
     
         33 . The composition of  claim 1  for the treatment of cancer. 
     
     
         34 . The composition of  claim 33 , wherein the cancer is selected from the group consisting of: cervical, vulvar, melanoma, breast, lung, ovarian, multiple myeloma, B cell lymphoma, hepatoma, sarcoma, bladder, prostate cancer, thyroid, H/N tumors, colon, rectal, renal, pancreatic, gastric, adenocarcinoma, T cell leukemia, lymphosarcoma, uterine, esophageal, non-Hodgkin's lymphomas, endometrial, and RCC tumors. 
     
     
         35 . The composition of  claim 1  for reducing tumor size. 
     
     
         36 . A method for treating cancer or inhibiting or preventing the growth or proliferation of cancer cells in a subject comprising administering the composition of  claim 1 . 
     
     
         37 . The method of  claim 36  wherein the cancer is selected from the group consisting of: cervical, vulvar, melanoma, breast, lung, ovarian, multiple myeloma, B cell lymphoma, hepatoma, sarcoma, bladder, prostate cancer, thyroid, H/N tumors, colon, rectal, renal, pancreatic, gastric, adenocarcinoma, T cell leukemia, lymphosarcoma, uterine, esophageal, non-Hodgkin's lymphomas, endometrial, and RCC tumors. 
     
     
         38 . The method of  claim 36 , further comprising the use of a topical composition comprising 1-isobutyl-1H-imidazo[4,5-c]quinolin-4-amine at the site of administration of said composition. 
     
     
         39 . A kit useful for treating cancer in a subject comprising a composition of  claim 1 , and instructions for its use thereof. 
     
     
         40 . The method of  claim 36 , wherein the subject is a mammal. 
     
     
         41 . The method of  claims 40 , wherein the mammal is a human. 
     
     
         42 . The composition of  claim 1 , wherein the liposome comprises a phospholipid. 
     
     
         43 . The composition of  claim 42 , wherein the phospholipid has at least one head group selected from phosphoglycerol, phosphoethanolamine, phosphoserine, phosphocholine and phosphoinositol. 
     
     
         44 . The composition of  claim 1 , wherein the liposome comprises unesterified cholesterol. 
     
     
         45 . The composition of  claim 1 , wherein the liposome comprises lipids in phospholipon 90 G. 
     
     
         46 . The composition of  claim 1 , wherein the liposome is obtained from archaebacterial bacteria. 
     
     
         47 . The composition of  claim 1 , wherein the liposome is obtained from natural lipids, synthetic lipids, sphingolipids, ether lipids, sterols, cardiolipin, cationic lipids and lipids modified with poly (ethylene glycol) and other polymers. 
     
     
         48 . The composition of  claim 45  wherein the synthetic lipids includes fatty acid constituents selected from lauroyl, myristoyl, palmitoyl, stearoyl, arachidoyl, oleoyl, linoleoyl, and erucoyl, or combinations thereof. 
     
     
         49 . A composition produced by a method comprising combining liposomes and an antigen capable of inducing a cytotoxic T lymphocyte (CTL) response with a carrier comprising a continuous phase of a hydrophobic substance. 
     
     
         50 . The composition according to  claim 49 , wherein said method further comprises combining a T helper epitope with said liposomes, said antigen and said carrier. 
     
     
         51 . The composition according to  claim 49 , wherein said method further comprises combining an adjuvant with said liposomes, said antigen and said carrier. 
     
     
         52 . The composition according to  claim 49 , wherein said antigen is encapsulated in said liposomes. 
     
     
         53 . The composition according to  claim 50 , wherein said T helper epitope is encapsulated in said liposomes. 
     
     
         54 . The composition according to  claim 51 , wherein said adjuvant is encapsulated in said liposomes. 
     
     
         55 . The composition according to  claim 49 , wherein said liposomes are freeze-dried before they are combined with said carrier. 
     
     
         56 . The composition according to  claim 50 , wherein said T helper epitope is a separate molecule from said antigen. 
     
     
         57 . The composition according to  claim 50 , wherein said antigen comprises said T helper epitope. 
     
     
         58 . The composition according to  claim 49 , wherein said antigen is an antigen as defined in  claim 6 . 
     
     
         59 . The composition according to  claim 49 , wherein said T helper epitope is a T helper epitope as defined in  claim 23 .