University of California

Other

Genteric, Inc.

Michael J Bennett

Michael H Nantz

Robert Wallace Malone

Rajiv P Balasubramaniam

Alfred M Aberle

Stephan S Rothman

The present invention describes methods for introducing nucleic acids into a target cell using a transition metal enhancer. A mixture containing nucleic acid and a transition metal enhancer is exposed to cells. The nucleic acid is taken up into the interior of the cell with the aid of the transition metal enhancer. Since nucleic acids can encode a gene, the method can be used to replace a missing or defective gene in the cell. The method can also be used to deliver exogenous nucleic acids operatively coding for proteins that are secreted or released from target cells, thus resulting in a desired biological effect outside the cell. Alternatively, the methods of the present invention can be used to deliver exogenous nucleic acids into a target cell that are capable of regulating the expression of a predetermined endogenous gene. This can be accomplished by encoding the predetermined endogenous gene on the nucleic acid or by encoding the nucleic acid with a sequence that is the Watson-Crick complement of the mRNA corresponding to the endogenous gene.

Method for nucleic acid transfection of cells

In the generation of cytofectin:polynucleotide complexes for transfection of cells, formulations, counterions, and reaction conditions for maximizing the transfection include using a cationic amine compound that has the general structure: ##STR1## wherein R.sub.4 and R.sub.5 are a pair of same or different lipoyl moieties selected from a group consisting of an alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, or alkynoyl groups and for R.sub.1, R.sub.2, and R.sub.3 at least two are hydroxylated, ether containing, or acyloxy containing alkyl, alkenyl, or alkynyl groups or at least one amine bonded halogen containing moiety selected from a group consisting of a halogenated alkyl, alkenyl, or alkynyl group or a mixture of at least one halogen containing moiety selected from a group consisting of a halogenated alkyl, alkenyl, or alkynyl group and at least one hydroxylated, ether containing, or acyloxy containing alkyl, alkenyl, or alkynyl group, and X.sup.- is an oxyanion or halide counterion.

Formulations and methods for generating active cytofectin:

A ball for warmup and therapy having a solid homogeneous body of molded elastomeric material of a size easily grasped in the palm of the hand, a weight of between 1-1.7 pounds, a hardness of 50-70 Shore A durometer and a percentage rebound of 65-90%. For therapy and warmup, the ball is generally used by grasping the ball in the palm of the hand and throwing or tossing this at a desired distance and angle, preferably using a full overhand motion, and catching the ball with any preferred hand as it bounces back to the user. This process is repeated as many times as the user is allowed, able or feels comfortable with.

Therapeutic ball

For use in transporting biologically active species into and through membrane barriers, a symmetrical cationic diamine compound having the general structure ##STR1## wherein m=1-10; R.sub.1 is a hydrogen, an alkyl group, an alkenyl group, a hydroxylated alkyl or alkenyl group, or an ether containing alkyl or alkenyl group; R.sub.2 is an alkyl group, an alkenyl group, or an alkyl or alkenyl containing acyl group; R.sub.3 is a hydrogen, an alkyl group, an alkenyl group, a hydroxylated alkyl or alkenyl group, or an ether containing alkyl or alkenyl group; R.sub.4 is a hydrogen, an alkyl group, an alkenyl group, a hydroxylated alkyl or alkenyl group, or an ether containing alkyl or alkenyl group; and X.sup.- is an anion.

Cationic transport reagents

For use in transporting biologically active species into and through membrane barriers, a cationic amine compound is utilized that has the general structure: ##STR1## wherein R.sub.4 and R.sub.5 are a pair of same or different lipoyl moieties selected from a group consisting of an alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, or alkynoyl groups and for R.sub.1 R.sub.2, and R.sub.3 at least two are hydroxylated, ether containing, or acyloxy containing alkyl, alkenyl, or alkynyl groups or at least one amine bonded halogen containing moiety selected from a group consisting of a halogenated alkyl, alkenyl, or alkynyl group or a mixture of at least one halogen containing moiety selected from a group consisting of a halogenated alkyl, alkenyl, or alkynyl group and at least one hydroxylated, ether containing, or acyloxy containing alkyl, alkenyl, or alkynyl group, and X.sup.- is an anion.

Polyfunctional cationic cytofectins

Amine containing compounds and their use in the generation of cytofectin:polynucleotide complexes for transfection of cells, formulations, counterions, and reaction conditions for maximizing the transfection include using cationic amine compounds that have the general structure: ##STR1## wherein R.sub.4 and R.sub.5 are a pair of same or different lipoyl moieties selected from a group consisting of an alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, or alkynoyl groups and for R.sub.1, R.sub.2, and R.sub.3 at least two are hydroxylated, ether containing, or acyloxy containing alkyl, alkenyl, or alkynyl groups or at least one amine bonded halogen containing moiety selected from a group consisting of a halogenated alkyl, alkenyl, or alkynyl group or a mixture of at least one halogen containing moiety selected from a group consisting of a halogenated alkyl, alkenyl, or alkynyl group and at least one hydroxylated, ether containing, or acyloxy containing alkyl, alkenyl, or alkynyl group, and X.sup.- is an oxyanion or halide counterion or ##STR2## wherein a, b, or d are the same or different and are from 0-10, usually between 0 and 3, preferably 0 or 1; 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 is halogen containing; and X.sup.- is an anion, usually an oxyanion or halide counterion.

Polyfunctional cationic cytofectins, formulations and methods for

For use transporting biologically active species into and through membrane barriers, a symmetrical cationic diamine compound having the general structure ##STR1##

For use in transporting biologically active species into and through membrane barriers, a symmetrical cationic diamine compound having the general structure ##STR1##

Growing community of inventors

Davis, CA, United States of America

Michael J Bennett