Johns Hopkins University

Other

University System of Maryland

Genta Incorporated

Paul O Ts'o

Paul S Miller

William A Carter

Jon J Hangeland

William G Nelson

Laure Aurelian

Yuan C Lee

Alan W Partin

Lionel N Simon

Stephen A Lesko

Zheng-Pin Wang

Tina Lynn Trapane

A method of treating HSV-2 infection which comprises administering to a host subject to said infection an effective amount of a polynucleotide of the formula polyrI.polyr(C.sub.12 U) where n is an integer from 4 to 29. The polynucleotide is advantageously administered intranasally.

Anti-viral and immuno stimulator polynucleotide duplex and use thereof

Acid resistant Oligomers suitable for oral administration, orally acceptable formulations of such Oligomers and preparation of pharmaceutical formations of such Oligomers are provided.

Oligonucleotide analogues having improved stability at acid pH

Oligodeoxynucleoside methylphosphonate neoglycopeptide conjugates and related compounds for tissue specific delivery of biologically stable, nonionic oligodeoxynucleoside analogs into cells.

Ligands to enhance cellular uptake of biomolecules

A method for enriching cancer cells such as prostate cancer cells in a bodily fluid sample is disclosed comprising (a) obtaining the sample comprising cancer cells and non-rare cells; (b) subjecting the sample to multiple density gradient separation comprising a first density gradient and a second density gradient, wherein the second density gradient is greater than the first density gradient, and producing a first fluid comprising an increased concentration of cancer cells of a first density, and a second fluid comprising an increased concentration of cancer cells of a second density, wherein the second density is greater than the first density; wherein subjecting the sample to multiple density gradient separation includes producing a plasma layer, a first interface layer, a first gradient layer, a second interface layer, a second gradient layer, and a cell pellet; wherein producing the first fluid includes combining the first interface layer and the first gradient layer and forming a first suspension; and wherein producing the second fluid includes combining the second interface layer and the second gradient layer and forming a second suspension; (c) subjecting said second fluid comprising the second suspension to a binding agent that binds non-rare cells; and (d) removing the bound non-rare cells from the second fluid to provide a second fluid enriched with the greater density cancer cells.

Method of enriching rare cells

A composition for inactivating a target nucleic acid which comprises an oligonucleoside alkyl or arylphosphonate analogue which is complementary to the sequence of the target nucleic acid and includes a functional group which reacts with the target nucleic acid to render the target nucleic acid inactive or nonfunctional.

Oligonucleoside alkyl or arylphosphonate derivatives capable of

Methods of detecting, recognizing or inhibiting or altering expression of a target sequence of a single stranded nucleic acid having any combination of purine and pyrimidine nucleosides by formation of triple helix complexes in conjunction with Second and Third Strands which comprise optionally covalently linked Oligomers are provided.

Formation of triple helix complexes of single stranded nucleic acids

A process for selectively controlling or interfering with the effect or function of foreign nucleic acid in the presence of otherwise normal living cells which comprises determining the base sequence for said nucleic acid and binding the said nucleic acid with an appropriately prepared nonionic oligonucleoside alkyl or arylphosphonate analogue which is complementary to the indicated sequence of the foreign nucleic acid.

Method for selectively controlling unwanted expression or function of

A nucleic acid-protein conjugate which is specific with respect to a selected living cell is prepared by linking said nucleic acid to a protein specific to said living cell.

Linkage of proteins to nucleic acids

A process for selectively controlling or interfering with the effect or function of foreign nucleic acid in the presence of otherwise normal living cells which comprises determining the base sequence for the nucleic acid and binding the nucleic acid with an appropriately prepared nonionic oligonucleoside alkyl or arylphosphonate analogue which is complementary to the indicated sequence of the foreign nucleic acid.

Process for selectively controlling unwanted expression or function of

Process for synthesizing deoxyribonucleoside methylphosphonates on polystyrene polymer supports which involves condensing 5'-dimethoxytrityldeoxynucleoside 3'-methylphosphonates. The oligomers are removed from the support and the base protecting groups hydrolyzed by treatment with ethylenediamene in ethanol, which avoids hydrolysis of the methylphosphonate linkages. Two types of oligomers are synthesized: those containing only methylphosphonate linkages, d-Np(Np).sub.n N, and those which terminate with a 5' nucleotide residue, dNp(Np).sub.n N. The latter oligomers can be phosphorylated by polynucleotide kinase, and are separated by polyacrylamide gel electrophoresis according to their chain length. Piperidine randomly cleaves the oligomer methylphosphonate linkages and generates a series of shorter oligomers whose number corresponds to the length of the original oligomer. Apurinic sites introduced by acid treatment spontaneously hydrolyze to give oligomers which terminate with free 3' and 5'-OH groups. These reactions may be used to characterize the oligomers.

Preparation of oligodeoxyribonucleoside alkyl or arylphosphonates

Oligonucleoside alkyl-- or aryl phosphonates are nonionic analogues of nucleic acid which possess unique physical and biological properties. These properties enable the analogues to enter living cells intact and to bind with specifically selected nucleic acids within the cell. As a result, the analogues can specifically inhibit the function or expression of a preselected nucleic acid sequence. Thus the analogues could be used to specifically inhibit the growth of tumor cells or replication of viruses in infected cells. Four methods are provided for preparing oligonucleoside methylphosphonates: (1) Coupling a protected nucleoside 3'-alkyl or aryl phosphonate with the 5'-hydroxyl group of a protected nucleoside using a condensing agent; (2) Coupling protected nucleoside 3'-alkyl or aryl phosphonic acid derivative with the 5'-hydroxy group of a protected nucleoside with the activated alkyl or aryl phosphonic acid derivative possessing functionalities which are good leaving groups; (3) Coupling a protected nucleoside 3'-alkyl or aryl phosphinate derivative with the 5'-hydroxyl group of a protected nucleoside with the resulting phosphinate derivative being then oxidized to the phosphonate; and (4) Converting a oligonucleoside methoxyphosphite derivative to the alkyl or aryl phosphonate derivative by reaction with an alkyl or aryl iodide. It has been demonstrated that procedures (1) and (2) can be used to prepare oligonucleoside methylphosphonates. Others have shown that procedure (4) can be used to prepare a diribonucleoside methylphosphonate.

Nonionic nucleic acid alkyl and aryl phosphonates and processes for

The present invention relates to the induction of interferon production in the cells of living organisms, including human beings. According to the invention, nucleic acid complexes, such as the polyriboinosinate and polycytidylate complex (rI.sub.n .multidot.rC.sub.n), are modified to yield unpaired bases, (uracil or guanine) along the polycytidylate strand which render the complexes more readily hydrolyzable by nucleases present in living cells. The modified complexes retain their ability to stimulate interferon release by the cells but are rendered more vulnerable to destruction within the cells, the modified complexes being significantly less toxic than the original complexes. In addition, polyinosinate strand now has been prepared to contain 5-16% 2'-O-methyl inosinate residues, designated as (rI.sub.5-20, 2'-MeI).sub.n. The new complex (rI.sub.5-20, 2'-MeI).sub.n .multidot.rC.sub.n, exhibits 100-fold more activity than rI.sub.n .multidot.rC.sub.n as an interferon inducer in human cells.

Induction of interferon production by modified nucleic acid complexes

The present invention relates to the induction of interferon production in the cells of living organisms, including human beings. According to the invention, nucleic acid complexes, such as the polyriboinosinate and polycytidylate complex (rI.sub.n.rC.sub.n), are modified to yield unpaired bases (uracil or guanine) along the polycytidylate strand which render the complexes more readily hydrolyzable by nucleases present in living cells. The modified complexes retain their ability to stimulate interferon release by the cells but are rendered more vulnerable to destruction within the cells, the modified complexes being significantly less toxic than the original complexes. In addition, polyinosinate strand now has been prepared to contain 5-16% 2'-O-methyl inosinate residues, designated as (rI.sub.5-20,2'-MeI).sub.n. The new complex (rI.sub.5-20,2'-MeI).sub.n.rC.sub.n, exhibits 100-fold more activity than rI.sub.n.rC.sub.n as an interferon inducer in human cells.

Growing community of inventors

Lutherville, MD, United States of America

Paul O Ts'o