Polyfunctional cationic cytofectins, formulations and methods for generating active cytofectin: polynucleotide transfection complexes

A cytofectin composition of matter having the structure:

• ##STR28## wherein m=1-10;
• R.sub.1, R.sub.2, and R.sub.3 are the same or different and are hydrogen, an alkyl group, an alkenyl group, an alkynyl group, a hydroxylated alkyl, alkenyl, or alkynyl group, an ether containing alkyl, alkenyl, or alkynyl group, or a trihalogenated alkyl, alkenyl, or alkynyl group, however, when two of the three are both hydrogens, both alkyls, or both alkenyls then the other is a trihalogenated group or when one of the three is hydrogen and another is an alkyl or alkenyl then the third is a trihalogenated group;
• R.sub.4 is an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group;
• R.sub.5 is an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group;
• and
• X.sup.- is a counterion.

A composition of matter having the structure:

• ##STR29## wherein n=1-10;
• R.sub.3 is a hydrogen, an alkyl group, an alkenyl group, an alkynyl group, or a hydroxylated alkyl, alkenyl, alkynyl group;
• R.sub.4 and R.sub.5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group;
• and
• X.sup.- is an anion.

A composition of matter having the structure:

• ##STR30## wherein n=1-10;
• R.sub.3 is a hydrogen, an alkyl group, an alkenyl group, an alkynyl group, or a hydroxylated alkyl, alkenyl, alkynyl group;
• R.sub.4 and R.sub.5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group;
• R.sub.6 is an alkyl group, an alkenyl group, an alkynyl group of from 1 to 10 carbons;
• R.sub.7 is an alkyl group, an alkenyl group, an alkynyl group of from 1 to 10 carbons;
• and
• X.sup.- is an anion.

A composition of matter having the structure:

• ##STR31## wherein a, b, or d are the same or different and are from 0-10;
• R.sub.4 and R.sub.5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group;
• R.sub.8, R.sub.9, or R.sub.10 are the same or different with each an alkyl, alkenyl, or alkynyl group or trihalogenated alkyl, alkenyl, or alkynyl group with at least one of the three being a trihalogenated group;
• and
• X.sup.- is an anion.

A composition of matter having the structure:

• ##STR32## wherein a=0-10;
• R.sub.4 and R.sub.5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group;
• R.sub.8 is a trihalogenated alkyl, alkenyl, or alkynyl group;
• R.sub.11 and R.sub.12 are the same or different with each an alkyl, alkenyl, or alkynyl group or trihalogenated alkyl, alkenyl, or alkynyl group;
• and
• X.sup.- is an anion.

A composition of matter having the structure:

• ##STR33## wherein a, b, or d are the same or different and are from 0-10;
• R.sub.4 and R.sub.5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group;
• R.sub.8, R.sub.9, or R.sub.10 are the same or different with each an alkyl, alkenyl, or alkynyl group or halogenated alkyl, alkenyl, or alkynyl group as long as one of the three is halogen containing;
• and
• X.sup.- is a counterion.

A cytofectin:polynucleotide complex produced by the steps comprising:

• a) combining a cytofectin with a lipid;
• b) mixing said combination of cytofectin and lipid with polynucleotide;
• and
• c) sonicating said mixture of polynucleotide, cytofectin, and lipid for a selected period of time at a predetermined temperature.