University of California

Megabios Corporation

Other

Valentis, Inc.

Biomics Biotechnologies, Co., Ltd.

Biocross Institute of Molecular Medicine (nantong) Co., Ltd.

Jinkang Wang

Francis C Szoka, Jr

Tiejun Li

YiXiang Lu

YunCheng Sun

Yilin Zhang

Li Chen

Ralph Niven

Pingzhong Huang

Yi-Lin Zhang

Yi Lin Zhang

Timothy D Heath

Alain Rolland

Igor Solodin

York YuanYuan Zhu

Hansj&ouml;rg Eibl

York Yuan Yuan Zhu

Yuhong Xu

This invention relates to the application of the highly conserved sequences of viral genome, especially from a highly conserved domain of enteroviral genome as templates to design target small ligand RNAs (sliRNAs). The resulting sliRNAs are therapeutically active ingredients in the treatment of the related diseases caused by pathological angiogenesis.

Application of highly conserved domain sequences from viral genome as template to design therapeutic sliRNAs

Application of highly conserved domain sequences from viral genome as template to design therapeutic slirnas

The invention comprises stabilized polynucleotide complexes that have a cryoprotectant and are lyophilized. Cryoprotectant compounds comprise carbohydrates or sugars, preferably lactose and sucrose, but also glucose, maltodextrins, mannitol, sorbitol, trehalose, and others. Other suitable cryoprotectants include amino acids such as betaines and prolines. Polynucleotide complexes stabilized according to the invention can be used for transfection, and exhibit improved tranfection efficiency with respect to polynucleotide complexes without cryoprotection.

Stabilized polynucleotide complexes and methods

Cationic lipids are provided which are useful in the preparation of liposomes and other lipid vesicle carriers. The lipids of the invention are particularly useful as carriers of nucleic acids and other negatively charged substances, for delivery to cells.

Cationic lipids

Methods for introducing nucleic acids into mammalian cells are provided which use imidazolium lipids. The imidazolium lipids have the formula:

Methods for introducing nucleic acids into mammalian cells using imidazolium lipids

Methods and compositions are provided for the introduction of polyanionic molecules, in particular, nucleic acids, into mammalian cells using certain phosphatidyl ethanolamines as helper lipids in conjunction with various cationic lipids. In particular, cationic lipid-mediated transfection of mammalian cells is improved by the use of lipid carriers comprising DLPE or DiPPE and cationic lipids.

Methods of delivery using cationic lipids and helper lipids

Compounds of the formula: ##STR1## are provided, in which R.sup.1 and R.sup.2 each independently represent a C.sub.8 -C.sub.24 saturated or unsaturated hydrocarbon chain, which is optionally interrupted by from 1 to 3 heteroatom moieties, such as --O--, --S--, --NH-- and --NR--. The symbol X represents --CH.sub.2 --, --O--, --S--, --NH-- or --NR--. The R group for each of the --NR-- moieties represents an alkyl group having from 1 to 4 carbon atoms. Finally, the subscript n represents the integer 1 or 2, and A.sup.- represents an anion, preferably chloride or citrate.

Compositions and methods using novel substituted imidazolium lipids

The invention separates defined, active complexes by a characteristic from Defined, active complexes that share a particular physicochemical characteristic such as density, surface charge or particle size are separated from complexes formed by the association of a polynucleotide with a transfecting component that increases transfection activity, such as a lipid, cationic lipid, liposome, peptide, cationic peptide, dendrimer or polycation. In a preferred embodiment, polynucleotide-transfecting component complexes are ultracentrifuged to resolve one or more bands corresponding to complexes having a specific polynucleotide-transfecting component interaction. Polynucleotide complexes having a cationic liposome transfecting component resolve into two primary bands corresponding to complexes formed either under excess lipid conditions or under excess polynucleotide conditions. In an alternate embodiment, polynucleotide-transfecting component complexes are resolved using cross-flow electrophoresis to identify complexes having specific interactions and to separate them from excess initial components.

Separation of active complexes

Cationic derivatives of biguanide are provided, which are useful in the preparation of lipid carriers for mediating transfection of mammalian cells in vivo and in vitro.

Amphiphilic biguanide derivatives

Polynucleotide complexes are stabilized by adding a cryoprotectant compound and lyophilizing the resulting formulation. The lyophilized formulations are milled or sieved into a dry powder formulation which may be used to deliver the polynucleotide complex. Delivery of the polynucleotide to a desired cell tissue is accomplished by contacting the tissue with the powder to rehydrate it. In a preferred embodiment, a dry powder formulation is used to transfer genetic information to the cells of the respiratory tract.

Growing community of inventors

San Francisco, CA, United States of America

Jinkang Wang