Ether lipids for hyperactivation of mammalian dendritic cells

A composition comprising an isolated ether lipid (ETL) of Formula (I):

• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5)2, —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• where R4 is H or (CH3)3N+—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof;
• and
• a TLR agonist.

A composition comprising an isolated ether lipid (ETL) of Formula (I):

• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• where R is H or (CH3)3N+—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof;
• and
• an antigen.

A composition comprising an isolated ether lipid (ETL) of Formula (I):

• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• where R4 is H or (CH3)3N−—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof;
• and
• dendritic cells.

A method for production of hyperactivated dendritic cells, the method comprising contacting the dendritic cells with a composition comprising effective amounts of an isolated ether lipid (ETL) of

• i) Formula (I):
• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• where R4 is H or (CH3)3N+—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof;
• or
• ii) Formula (IV-F):
• wherein R2 is H, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C21-C24 n-alkyl;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a salt thereof;
• and
• a TLR7/8 agonist for production of hyperactivated dendritic cells, wherein the hyperactivated dendritic cells secrete IL-1beta without undergoing pyroptosis.

A method of preparing an immunogenic composition, the method comprising:

• a) depleting leukocytes from a suspension of cells prepared from a tumor to obtain a tumor cell-enriched suspension;
• b) lysing cells from the tumor cell-enriched suspension to obtain a tumor cell lysate;
• and
• c) contacting the tumor cell lysate with an isolated ether lipid (ETL) of
• i) Formula (I):
• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• where R4 is H or (CH3)3N+—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof;
• or
• ii) Formula (IV-F):
• wherein R2 is H, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C21-C24 n-alkyl;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a salt thereof;
• and
• a toll-like receptor (TLR) agonist to obtain the immunogenic composition.

A method of treating cancer, the method comprising:

• a) preparing an immunogenic composition comprising a tumor cell lysate, an isolated ether lipid (ETL) of
• i) Formula (I):
• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C(H5);
• R3 is C13-C24 n-alkyl;
• where R4 is H or (CH3)3N+—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof:
• or
• ii) Formula (IV-F):
• wherein
• R2 is H, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C21-C24 n-alkyl;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a salt thereof;
• and
• a toll-like receptor (TLR) agonist, wherein the tumor cell lysate is or has been prepared from a sample of a tumor obtained from the mammalian subject with cancer;
• and
• b) administering to the subject an effective amount of the immunogenic composition.

A composition comprising an isolated ether lipid (ETL) of:

• i) Formula (I):
• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• where R4 is H or (CH3)3N+—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof;
• or
• ii) Formula (IV-F):
• wherein R2 is H, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C21-C24 n-alkyl;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a salt thereof;
• and
• a pathogen recognition receptor (PRR) agonist.

A pharmaceutical formulation comprising an isolated ether lipid (ETL) of

• i) Formula (I):
• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• where R4 is H or (CH3)3N−—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof;
• or
• ii) Formula (IV-F):
• wherein R2 is H, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C21-C24 n-alkyl;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a salt thereof;
• and
• a pharmaceutically acceptable excipient.

A composition for hyperactivation of human dendritic cells, comprising an isolated ether lipid (ETL) of

• i) Formula (I):
• wherein:
• R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• where R4 is H or (CH3)3N+—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a pharmaceutically acceptable salt thereof;
• or
• ii) Formula (IV-F):
• wherein R2 is H, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C21-C24 n-alkyl;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a salt thereof;
• and
• a pathogen recognition receptor (PRR) agonist, wherein the alkyl chain is a C22 n-alkyl chain, and wherein the composition is effective for achieving a higher level of dendritic cell hyperactivation than a comparator composition comprising PGPC in place of the ETL.

An isolated ether lipid (ETL) of Formula (I):

• wherein R1 is H or
• R2 is H, C1-C4 alkyl, —(C═O)—NH2, —(C═O)—NH(R5), —(C═O)—N(R5)2, or —CH2—C6H5;
• R3 is C13-C24 n-alkyl;
• wherein R4 is H or (CH3)3N+—(CH2)2—;
• and
• each R5 is independently C1-C4 alkyl;
• or a protonated or deprotonated form thereof;
• or a salt thereof.

A compound of Formula (III-A-1):

• wherein:
• R2 is —(C═O)—NH2, —(C═O)—NH(R5), or —(C═O)—N(R5)2;
• R3 is C21-C24 n-alkyl;
• and
• each R5 is independently C1-C4 alkyl;
• or a pharmaceutically acceptable salt thereof.