Novel Compositions for the Delivery of Negatively Charged Molecules

Abstract

This invention features permeability enhancer molecules, and methods, to increase membrane permeability of various molecules, such as nucleic acids, polynucleotides, oligonucleotides, enzymatic nucleic acid molecules, antisense nucleic acid molecules, 2-5A antisense chimeras, triplex forming oligonucleotides, decoy RNAs, dsRNAs, siRNAs, aptamers, or antisense nucleic acids containing nucleic acid cleaving chemical groups, peptides, polypeptides, proteins, carbohydrates, steroids, metals and small molecules, thereby facilitating cellular uptake of such molecules.

Description

FIELD OF THE INVENTION[0001]This patent application is a continuation of Beigelman et al., U.S. Ser. No. 09 / 120,520, filed Jul. 21, 1998, entitled “NOVEL COMPOSITIONS FOR THE DELIVERY OF NEGATIVELY CHARGED MOLECULES”, which claims priority from Beigelman et al., U.S. Ser. No. 60 / 053,517, filed Jul. 23, 1997, entitled “NOVEL COMPOSITIONS FOR THE DELIVERY OF NEGATIVELY CHARGED MOLECULES” and Beigelman et al., U.S. Ser. No. 60 / 072,967, filed Jan. 29, 1998, entitled “NOVEL COMPOSITIONS FOR THE DELIVERY OF NEGATIVELY CHARGED MOLECULES”. These applications are hereby incorporated by reference herein in their entirety including the drawings.BACKGROUND OF THE INVENTION[0002]The following is a brief description of the delivery of biopolymers. This summary is not meant to be complete but is provided only for understanding of the invention that follows. This summary is not an admission that all of the work described below is prior art to the claimed invention.[0003]Trafficking of large, charge...

AI Technical Summary

Benefits of technology

[0013]This invention features lipid-based compositions, such as cationic lipid-based compositions, that facilitate delivery of molecules into a biological system, for example into cells such as mammalian cells. The present invention discloses the design, synthesis, and cellular testing of novel agents for the delivery of various molecules, for example nucleic acids, polynucleotides, oligonucleotides, and / or negatively charged molecules, in vitro and in vivo. Also disclosed are screening procedures for identifying the optimal delivery vehicles for any given nucleic acid and cell type. In general, the transporters or cytofectins described here are designed to be used either individually or as part of a multi-component system. Examples of such multi-component systems comprising lipid compounds of the invention are shown in Tables II, VI, VII, VIII, and IX. The lipid compounds of the invention generally shown in FIG. 1 and Table X, are expected to improve delivery of nucleic acids, polynucleotides, oligonucleotides, and / or negatively charged molecules, into a number of cell types originating from different tissues, in the presence or absence of serum.

[0035]In another embodiment, multi-domain cellular transport vehicles (MCTV) including one or more lipids of formula I-IX and / or Lipid ID Nos: 700, 701, 705, 709, 719, 722, 723, 725, 726, 727, 732, 736, 737, 738, 739, 742, 743, 744, 745, 746, 747, 749, 750, 751, 752, and / or 753, that enhance the cellular uptake and transmembrane permeability of various molecules, for example, nucleic acids, polynucleotides, oligonucleotides and / or negatively charged molecules in a variety of cell types are provided. Examples of such lipids are shown in Table X. The lipids of the invention are used either alone or in combination with other compounds with a neutral or a negative charge including but not limited to neutral lipid and / or targeting components, to improve the effectiveness of the lipid formulation in delivering and targeting molecules such as nucleic acids, polynucleotides, oligonucleotides, or negatively charged polymers to cells. In addition, these delivery vehicles can be used to increase the transport of other impermeable and / or lipophilic compounds into cells.

[0038]In yet another embodiment, the lipid molecules of the invention, such as cationic lipids, are provided as a lipid aggregate, such as a liposome, and co-encapsulated with the compound or polymer to be delivered. Liposomes, which can be unilamellar or multilamellar, can introduce encapsulated material into a cell by different mechanisms. See, Ostro, Scientific American 102, January 1987. For example, the liposome can directly introduce its encapsulated material into the cell cytoplasm by fusing with the cell membrane. Alternatively, the liposome can be compartmentalized into an acidic vacuole (i.e., an endosome) having a pH below 7.0. This low pH allows ion-pairing of the encapsulated enhancers and the negatively charged polymer, which facilitates diffusion of the enhancer:polymer complex out of the liposome, the acidic vacuole, and into the cellular cytoplasm.

Problems solved by technology

Trafficking of large, charged molecules into living cells is highly restricted by the complex membrane systems of the cell.

Viral vectors can be used to transfer genes efficiently into some cell types, but they cannot be used to introduce chemically synthesized molecules into cells.

Synthetic nucleic acids as well as plasmids may be delivered using the cytofectins, although their utility is often limited by cell-type specificity, requirement for low serum during transfection, and toxicity.

However, each formulation is of limited utility because it can deliver plasmids into some but not all cell types, usually in the absence of serum (Bailey et al., 1997, Biochemistry, 36, 1628).

However, the number of available delivery vehicles that may be utilized in the screening procedure is highly limited, and there continues to be a need to develop transporters that can enhance nucleic acid entry into many types of cells.

Images