CA2071936A1 - Boronated nucleosides - Google Patents
Boronated nucleosidesInfo
- Publication number
- CA2071936A1 CA2071936A1 CA002071936A CA2071936A CA2071936A1 CA 2071936 A1 CA2071936 A1 CA 2071936A1 CA 002071936 A CA002071936 A CA 002071936A CA 2071936 A CA2071936 A CA 2071936A CA 2071936 A1 CA2071936 A1 CA 2071936A1
- Authority
- CA
- Canada
- Prior art keywords
- boronated
- nucleoside
- cyanoborane
- borane
- group
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D473/00—Heterocyclic compounds containing purine ring systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D473/00—Heterocyclic compounds containing purine ring systems
- C07D473/02—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6
- C07D473/18—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6 one oxygen and one nitrogen atom, e.g. guanine
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D473/00—Heterocyclic compounds containing purine ring systems
- C07D473/26—Heterocyclic compounds containing purine ring systems with an oxygen, sulphur, or nitrogen atom directly attached in position 2 or 6, but not in both
- C07D473/32—Nitrogen atom
- C07D473/34—Nitrogen atom attached in position 6, e.g. adenine
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic System
- C07F5/02—Boron compounds
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
- C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H23/00—Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12
Abstract
A novel class of pharmaceutically active boronated nucleosides are provided. The nucleosides are boronated at a ring nitrogen of the purine or pyrimidine or analogues thereof. Also provided are phosphate esters of these nucleosides and oligomers thereof.
Methods of making and using the boronated nucleosides are also disclosed.
Methods of making and using the boronated nucleosides are also disclosed.
Description
~u . ~J.,i~
BORONATED NUCLEOSIDES
Field of the Invention This invention pertains to novel Boron containing compounds having pharmaceutical activity.
More specifically, compounds of the present invention include nucleoside analogues having antineoplastic activity. These compounds seemingly function as antimetabolites with additional utility in Boron Neutron Capture Therapy (BNCT).
Introduction Antimetabolites are a well known class of antineoplastic agents that function by interfering with -nucleic acid sythesis and consequently, replication within the target cells. Some of these compounds structurally mimic biosynthetic precursors of the naturally occurring nucleic acids, which are essential for replication and cell survival. By replacing these precursors, but without exhibiting the same biological effect, these compounds disrupt replication resulting in the demise of the the target cell.
Many antimetabolites have significant antiviral and antitumor activity and are incorporated in a variety of therapeutic regimens. But despite the therapeutic benefits of such compounds, their use is ~' , . ,...',,~,,, ' :' . ',','. -.
:: : . ' : , ' :.
~' :.. ' ' .' . . -2071 '~fi often acco~panied by deleterious side effects, e.g.
nausea, alopecia, and bone marrow depression.
Accordingly, a great deal of in~erest has focused on synthesizing new analogues with improved therapeutic indexes.
We have recently discovered that boron containing nucleotides may be one class of improved nucleic acid analogues. Some exemplary boronated nucleotides are described in copending, commonly owned U.S. patent application Serial No. of B.
Spielvogel, A. Sood, I. Ilall, and B. Ramsay-Shaw titled "Oligoribonucleoside and Oligodeoxyribonucleoside Boranophosphates~' and filed November 30, 1989, which is incorporated herein by reference. There we describe, lS for example, boronated oligonucleotides that contain a boron functionality attached to internucleotide phosphorus.
Boron containing compounds are also useful in an antineoplastic regimen known as Boron Neutron Capture Therapy (BNCT). Soloway, A.H., Prog~ess in :
Boron Chemistry; Steinberg, H., McCloskey, A. L., Eds.;
the Macmillan Company: New York, 1964; Vol. 1, Chapter 4, 203-234. BNCT requires two components (Boron-10 and low energy thermal neutrons) for a radiotoxic reaction.
The inherent advantage is that each component can be manipulated independently to produce the desired radiation effect. Boron-10 has a high cross section for thermal neutrons and after neutron capture, the particles generated, Li & ~, are relatively large by radiation standards and thus have a relatively short track in tissue, 10-14 microns. The Boron-10 is non-radioactive and for use in BNCT, its compounds do not have to be cytotoxic towards tumor cells. Thermal neutrons have such low energy that they cannot ionize tissue components per se. Upon neutron capture, however, the energy generated is sufficient to destroy the cell. The problem in making this procedure :. : :
. .
- - :. . ~ ., , , " , , ~
v clinically effective has stemmed not from the concept, per se, but from lack of knowledge in medicinal chemistry, nuclear engineering and physics, immunology, physiology and pharmacology. The present invention arose from our continued research on new boron-containing compounds having pharmaceutical activity.
8ummary of the Invention A first aspect of the present invention is a boronated nucleoside comprising a nucleoside which is lo N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -~H3, and -BH2COOR, wherein R is Cl to C18 alkyl. Preferably, R is Cl to C9 alkyl, and most preferably R is Cl to C4 alkyl.
A second aspect of the present invention is a boronated nucleotide comprising a 5' phosphate ester of a boronated nucleoside as described above.
A third aspect of the present invention is a boronated oligonucleotide comprising a chain of natural or modified ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a boronated nucleotide as described above.
Nucleosides of the present invention have phar~aceutical activity, including antihyperlipidemic, antiinflammatory, and antineoplastic activity.
Nucleotides of the present invention are useful as intermediates for making oligonucleotides of the present invention. Oligonucleotides of the present .
invention are useful as antisense agents and probes.
A method for synthesizing N-boronated nucleosides of the present invention from a substrate nucleoside, which substrate nucleoside is comprised of a ribose moiety covalently bound to a nitrogenous base, is also disclosed. The method comprises:
(a) protecting the hydroxy substituents of the ribose; then , :. ,. . - : .:
: ~
~n7~
(b) boronating the nitrogenous base; and then (c) deprotecting the hydroxy substituents.
Preferably, the nitrogenous base is boronated by 5 reacting the substrate nucleoside with either polymeric BH2CN or LX, wherein L is a Lewis base and X is a boron-containing substituent selected from the group consisting of -BH2CN, -BH" and -B)~2COOR, wherein R is as given above.
The term alkyl, as used herein, refers to linear or branched alkyl groups which may be saturated or unsaturated. Preferred alkyl groups are linear and saturated.
Detailed Description of the Invention We have successfully synthesized novel boronated nucleosides that have surprising antineoplastic, antiinflammatory and antihyperlipidemic properties. The present invention provides novel pharmaceutical agents, methods for their synthesis, methods for treating patients in need of such treatment, and pharmaceutical formulations comprising such agents.
The nucleosides of the present invention comprise nucleosides having a boron-containing substituent on the base. More specifically, the present invention provides N-boronated nucleosides wherein the boronated nitrogen is a ring nitrogen of the base covalently bonded to the boron of the boron-containing substituent. The base is a purine or a pyrimidine or an analog thereoE (as discussed in detail below). A preferred group of bases are those selected from the group consisting of adenine, cytosine, guanine, inosine, and the analogs thereof.
By nucleosides we mean a purine or pyrinidine base, and analogs thereof, linked to a pentose. The pentose is preferably D-ribose, 2'-deoxy-D-ribose or 2',3'-dideoxy-D-ribose. Thus the nucleosides of the . ' ' ' ' , . ' . . .
~ n ~
~ U . _ L~ J
present invention include both naturally occurrin~
nucleosides and analogs thereof.
In the case of a purine nucleoside, boronation occurs preferentially at either the Nl or N7 position depending upon the purine. For example, when the purine is adenine, boronation occurs at the Nl position. In the case of guanine, boronation occurs at the N7 position. It is contemplated that boronation at other nitrogens on the nitrogenous bases will yield lo additional useful agents.
The nucleoside base may generally be a natural base, such as adenine, thymine, cytosine, guanine, uracil, xanthine, or hypoxanthine, (the latter two being the natural degradation products) or an analog thereof as found in, for example, 4-acetylcytidine; 5-(carboxyhydroxyl- methyl)uridine; 2'-0-methylcytidine; 5-carboxymethyl- aminomethyl-2-thiouridine; 5-carboxymethylaminomethyl-uridine;
dihydrouridine; 2'-0-methylpseudo-uridine; beta,D-galactosylqueosine; 2'-0-methylguanosine; N6-isopentenyladenosine; l-methyladenosine; l-methyl-pseudo-uridine; l-methylguanosine; l-methylinosine;
2,2-dimethylguanosine; 2-methyladenosine; 2-methyl-guanosine; 3-methyl-cytidine; S-methylcytidine; N6-methyladenosine; 7-methyl-guanosine; 5-methylamino-methyluridine; 5-methoxyamino-methyl-2-thiouridine;
beta,D-mannosylqueosine; 5-methoxy-carbonylmethyl-uridine; 5-methoxyuridine; 2-methylthio-N6-isopentenyl-adenosine; N-((9-beta-D-ribofuranosyl-2-methylthio-purine-6-yl)carbamoyl)threonine; N-((9-beta-D-ribofuranosyl- purine-6-yl)N-methyl-carbamoyl)-threonine; uridine-5-oxyacetic acid methylester;
uridine-5-oxyacetic acid (v); pseudouridine; queosine;
2-thiocytidine; 5-methyl-2-thiouridine; 2-thiouridine;
4-thiouridine; 5-methyluridine; 2'-O-methyl-5-methyluridine; and 2'-O-methyluridine. Preferred analogs are the methylated analogs.
.
.~ .
~o J ~u~J
Exemplary boronated nucleosides of the present invention are:
(A) ~uanosine-N7-cyanoborane;
(B) inosine-N7-cyanoborane;
(C) adenosine-Nl-cyanoborane;
(D) cytidine-N3-cyanoborane;
(E) guanosine-N7-borane;
(F) inosine-N7-borane;
(G) adenosine-Nl-borane; .-:
(T~) cytidine-N3-borane;
(I) guanosine-N7-carbomethoxyborane;
(J) inosine-N7-carboethoxyborane;
(K) adenosine-Nl-carbopropoxyborane;
(L) cytidine-N3-carbobutoxyborane;
(M) 2'-deoxyguanosine-N7-cyanoborane;
(N) 2'-deoxyinosine-N7-cyanoborane;
(0) 2'-deoxyadenosine-Nl-cyanoborane;
(P) 2'-deoxycytidine-N3-cyanoborane;
(Q) 2'-deoxyguanosine-N7-borane;
(R) 2'-deoxyinosine-N7-borane;
(S) 2'-deoxyadenosine-Nl-borane;
~T) 2'-deoxycytidine-N3-borane;
(U) 2'-deoxyguanosine-N7-carbomethoxyborane;
(V) 2'-deoxyinosine-N7-carboethoxyborane;
(W) 2'-deoxyadenosine-N1-carbopropoxyborane;
(X) 2'-deoxycytidine-N3-carbobutoxyborane; ~ .
(Y) 2',3'-dideoxyguanosine-N7-cyanoborane;
(Z) 2',3'-dideoxyinosine-N7-cyanoborane;
(AA) 2',3'-dideoxyadenosine-Nl-cyanoborane;
(A3) 2',3'-dideoxycytidine-N3-cyanoborane;
(AC) 2',3'-dideoxyguanosine-N7-borane;
(AD) 2',3'-dideoxyinosine-N7-borane;
(AE) 2',3'-dideoxyadenosine-Nl-borane;
(AF) 2',3'-dideoxycytidine-N3-borane; -(AG) 2',3'-dideoxyguanosine-N7-carbomethoxyborane;
.
.:,. - - : . :
: , - ',: . .~ ~ ' - ' : , , ' ' :, . .
.
207~9~fi (AH) 2',3'-dideoxyinosine-N7-carboethoxyborane;
tAI) 2',3'-dideoxyadenosine-~11-carbopropoxyborane; and S tAJ) 2~3~-dideoxycytidine-N3 carbobutoxyborane.
Particularly preferred boronated nucleosides are: 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2'-deoxyadenosine-Nl-cyanoborane, and lo 2'-deoxycytidine-N3-cyanoborane.
The nucleosides of the present invention further comprise 5'-phosphate esters of the N-boronated nucleosides described herein, such phosphate esters are also known as nucleotides. Such nucleotides, particularly the monophosphates, are protected in conventional manner and used for the synthesis of oligonucleotides, as discussed below. Such phosphate esters include 5' mono-, di- and triphosphates, which may be protected as esters. Additionally, molecules and macromolecules comprising multimers of two or more nucleosides, which may be linked via a 3'-5' phosphate ester, e.g. oligonucleotides (the terms "oligonucleotides~ and "polynucleotides~ being used interchangeably herein), and comprising one or more N-boronated nucleosides are also the subject of the present invention. Accordingly, N-boronated nucleotides, oligonucleotides, and polynucleotides may be used as therapeutic agents and otherwise useful reagents, e.g. diagnostic reagents.
Oligonucleotides of the present invention can be synthesized in accordance with methods that are well known in the art. Such methods include the phosphite method and the phosphotriester method. 1 Chemistry of Nucleosides and Nucleotides, 294ff (L. Townsend ed.
1988). ~he length of the oligonucleotide is not :i critical, as modern synthetic techniques and splicing techniques have made synthetic oligonucleotides of :. . ' .,;, . : , , .
~ u J ~
considerable length feasible. Thus, the oligonucleotide may for example be 2 to 3 nucleotides long, 2 to 18 nucleotides long, 2 to 30 nucleotides long, 2 to 50 nucleotides long, or 50 or more S nucleotides long.
Oligonucleotides containing N-boronated bases may alternatively be prepared, with boronation occur~ng randomly, in essentially the same manner as the nucleoside, but with an oligonucleotide substituted for lo the nucleoside. For example, in such a reaction, the 3' terminus of the oligonucleotides may be immobilized to a solid support (e.g., controlled pore glass), the 5' terminus protected as the dimethyltrityl ether, and amino groups on bases protected with isobutyryl groups.
Oligonucleotides used for such a reaction preferably contain at least one base which is not thymidine or a thymidine analog, as these are less preferred bases for boronation.
Derivatives of the oligonucleotides and polynucleotides may additionally be formed by modifying the internucleotide phosphodiester linkage.
Internucleotide phosphodiester linkages in the chain are modified, for example, to the methylphosphonate, the phosphotriester, the phosphorothioate, the phosphorodithioate, and the phosphoramidate, all as is known in the art.
Additional synthetic analogues of the nucleosides, nucleotides, and oligonucleotides of the present invention may be formed by otherwise modifying the 3' or 5' end of the nucleoside, and any 2' hydroxyl groups. Groups that can be added to the 3' or 5' end vary widely, from relatively inert protecting groups to 1~^
reactive groups or groups with special properties to obtain desireable physical, chemical, or biochemical effects.
A wide variety of protecting groups can be substituted on the 2', 3', and 5' hydroxy groups, such .
!
207193~
as the triphenylmethyl (trityl) group and its derivatives on the 5' hydroxy group, and acetyl, ~enzo~l, or the 3'-O-succinyl group on the 3' hydroxy gro~p, as is known in the art. See 1 Chemistry of rJucleosides and Nucleotides, 287-92 (L. Townsend ed.
1988). In general, the 5' hydroxy group is protected with an acid labile group and the 3' hydroxy group is protected with an acyl group. Id. at 289 (When the 5' hydroxyl group is protected with an acid labile group lo such as mono- and dimethoxytrityl, the 3'-hydroxyl group of deoxynucleosides can be protected with acyl groups.). ~n general, a 2' hydroxy group is protected as a methyl ether, protected with a silyl group, or the 2' and 3' hydroxy groups may be protected together as an acetal.
Reactive groups or groups with special properties may also be attached at the 3' or 5' position. For example, analogs may be formed by joining an intercalating agent to oligonucleotides and polynucleotides in the manner described in U. S. Patent No. 4,835,263 to Nguyen et al. (the disclosure of this and all other patent references cited herein is incorporated herein by reference).
The present invention also provides methods for preparing the compounds of the present invention.
The boronation of a nucleoside is accomplished by first preparing a hydroxy- protected nucleoside. Protecting groups and methods for their use are well known in the art. See, Carey and Sundberg, Advanced Orqanic Chem i try. Part B, pp. 408-414 (1980). Preferred protecting groups are organosilanes, e.g., chlorotriisopropyl silane will form the triisopropyl silyl ether.
As an example, the nucleoside is 0-protected by forming a silyl ether by reaction with excess chlorotriisopropyl-silane at room temperature in the presence of imidazole. The exchange reaction is ' ' ' ' ' `' : .. , ::
effected by reacting the O-protected nucleoside with an organoborane. The organoborane is generally either polvmeric B~2C~ or a compound LX, wherein L is a Lewis base and X is a boron-containing su~stituent as given above. Suitable Lewis bases include amine, phosphine, sulfide, and ether (e.g., tetrahydrofuran). The strength and steric properties of the Lewis base should be chosen so as to provide a suitable leaving group.
Exemplary organoboranes include aniline-cyanoborane, 1~ triphenylphosphine-carboalkoxyboranes (wherein the alkoxy group alkyl is R as given above), dimethylsulfide-borane, and tetrahydrofuran-borane. A
preferred orqanoborane is triphenylphosphine-cyanoborane, in which case the resulting product is an O-protected ribonucleoside N-cyanoborane. The exchange reaction is effected by reaction in anhydrous tetrahydrofuran (THF) at reflux temperature using two ~ -equivalents of triphenylphosphine-cyanoborane. The reaction is at equilibrium in two to three hours.
The resulting O-protected N-boronated ribonucleoside is deprotected by appropriate methods -known in the art. For example, when protected by formation of a silyl ether, deprotection can be effected under hydrolytic or nucleophilic conditions.
In our preferred case the silyl ether is effectively removed under hydrolytic conditions in the presence of fluoride ion, e.g., tetra-n-butylammonium fluoride.
The compounds of the present invention have pharmaceutical activity and are useful in treating mammals (e.g., human, cat, dog, cow, horse, mouse) suffering from one or more of several maladies. These compounds show pharmaceutical activity in killing cancer cells in_vitro, and may be useful in combatting corresponding tumors in vivo. For example, the compounds of the present invention show cytotoxic activity against colorectal carcinoma, adenocarcinoma, osteosarcoma, breast carcinoma and glioma.
.. .
. .
Accordingly, the compounds of the present invention provide a method for treating a tumor bearing mammal comprising administering to said mammal a therapeutically effective amount of a boronated S nucleoside of the present invention. Furthermore, it is contemplated that the antineoplastic efficacy of these compounds can be improved or supplemented by the conjoint administration with other known antineoplastic agents, as, for example, in a combination chemotherapy lo regimen. Exemplary of such other known antineoplastic agents are: vinca alXaloids such as vincristine, vinblastine, and vindesine; epipodophyllotoxins such as etoposide and teniposide; anthracycline antibiotics such as daunorubicin, doxorubicin, mitoxantraone, and bisanthrene; actinomyGin D; and plicamycin.
In addition to the direct inhibition of tumor growth, the preferential localization of boron compounds in the neoplasm of tumor cells will allow the use of boron-10 neutron capture therapy (BNTC) for the ~ -destruction of tumor cells. Moreover, the dual effect of this therapeutic regimen may lower the therapeutically effective amounts of the pharmaceutically active agents, and thereby reduce the ~-deleterious side effects that often accompany the use of such agents. Thus, the present invention also provides methods for treating tumor-bearing mammals in which the mammal is administered a boronated nucleoside as described herein and then exposed to thermal neutron radiation. The thermal neutron radiation is administered in an amount and in a manner effective for 10B located in a tumor by virtue of the administration of the compound of the present invention to the subject to capture a neutron, decay, and release an alpha particle in cells of the tumor.
The compounds of the present invention also have pharmaceutical activity as antiinflammatory agents in mammals. The compounds of the present invention : .
~v ,i~
provide a method for treating a mammal suffering from inflammation comprising administering to said mammal a therapeutically ef~ective amount of an N-boronated nucleoside. The compounds o~ the present invention may provide additional utility when conjointly administered with other known antiinflammatory agents or pain killers or some such pharmaceutical. Exemplary of other known antiinflammatory agents are acetylsalicylic acid (asprin), salicylic acid, diflunisal, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine, dipyrone, apazone, acetaminophen, indomethacin, sulindac,meclofenamate, tolmetin, zomepirac, ibuprofen, and piroxicam.
The compounds of the present invention are useful as hypolipidemic agents. The compounds of the present invention provide a method for treating a mammal suffering from hyperlipidemia comprising administering to said mammal a therapeutically effective amount of an N-boronated nucleoside.
Additionally, the compounds of the present invention provide a method for treating a mammal suffering from hypercholesterolemia comprising administering to said mammal a therapeutically effective amount of an N- -boronated nucleoside. ~y administering these compounds to hyperlipidemic patients the total lipoprotein level may be reduced or the lipoprotein profile may be improved. Furthermore, these compounds may be con~ointly administered with other known hypolipedemic agents to enhance or supplement their efficacy.
Exemplary of such other ~nown hypolipidemic agents are nicotinic acid, clofibrate, gemfibrozil, probucol, cholestyramine, colestipol, compactin, mevinolin, choloxin, neomycin, and beta-sitosterol.
The compounds of the present invention may be administered in any of the variety of modes currently employed for analogous pharmaceutical agents, which modes are well known in the art. For example, these .
` ~
.
~ n ~
compounds may be administered systemically. Systemic administration includes parenteral administration and gastro-enteral administration.
When prepared in a pharmaceutical formulation for parenteral administration the compounds of the present invention should be prepared in a pharmaceutically acceptable carrier such as substantially non-pyrogenic, sterile, parenterally acceptable, aqueous solutions.
Alternatively, the compounds of the present invention may be formulated in pharmaceutical preparations for gastro-enteral administration. Such pharmaceutical preparations include tablets, capsules and suppositories. When formulated for administration according to any of the above methods the pharmaceutical preparations may further comprise buffers, binders, and other pharmaceutically acceptable excipients as are well known in the art.
A therapeutically effective amount of a boronated nucleoside is in the range of about 0.1-100 mg/kg/day. The preferred range is about 0.5-50 mg/kg/day. More preferred is an amount in the range of about 1-10 mg/kg/day. When administered conjointly with other pharmaceutically active agents even less of the boronated nucleoside may be therapeutically effective.
The oligonucleotides of the present invention may be used as probes in a variety of diagnostic techniques. One such diagnostic technique is Magnetic Resonance Imaging (MRI). MRI is a noninvasive technique used to detect the presence and location of tumors in a patient. For example, as contemplated in the present context, cancer cell specific boronated compounds are administered to a patient, whereupon they concentrate in cancerous tissue. The MRI instrument is capable of detecting and locating regions of abnormal concentrations of Boron. By indicating the regions .. . . . . ...
: . :.......... . ~
; . ~
2'~ n~
~J ~ ., V .J
havinq high relative concentrations of Boron, MRI
establ ishes the presence and location of tumors.
Another diagnostic application of the oligonucleotides of the present invention is their use as molecular probes. By incorporating N-boronated nucleosides, or their 5'-phosphate esters, into an oligonucleotide, either at an interior or terminal position, a detectable oligonucleotide probe is constructed that can be used to detect the presence of complementary sequences of DNA or RNA in a sample.
These probes can be used in any suitable environment, such as Southern blotting and Northern blotting, the details of which are known. See e.g., R. Old and S. Primrose, Principles of Gene Manipulation, 8-10 (3d Ed. 1985). When used as probes, the boron atom serves as a radiolabel, though it is not itself radioactive until exposed to thermal neutron radiation (low energy neutrons). When exposed to low energy neutrons, IB absorbs a neutron and forms IlB, which rapidly decays and releases an alpha particle, thus providing a detectable signal. The techniques involved in the generation of the alpha particle are known. See. e.~., A. Soloway, Borax Rev. 3, 7-9 (1988).
Oligonucleotides of the present invention which are capable of binding to polyribonucleic acid or polydeoxyribonucleic acid are useful as antisense agents in the same manner as conventional antisense agents. See gene~allv Antisense Molecular Biology and S-oligos, Syntkesis 1 (Oct. 1988) (published by Synthecell Corp., Rockville, MD); 2 Discoveries in Antisense Nucleic Acids (C. Brakel and R. Fraley eds.
1989). Antisense agents of the present invention may be used by contacting an antisense agent which is capable of selectively binding to a predetermined polydeoxyribonucleic acid sequence or polyribonucleic acid sequence to a cell containing such sequence (e.g., .
. . . . .
.: . . -; . , . .
: . . ., ~.: . ;
.. . : . ~ -. : .
' ::
:. , :.. :: . ...
9 0 / ~ 7 4 4 6 -15- lP ~ Us 0 6 JAN 1992 by adding the antisense agent to a culture medium containing the cell) so that the antisense agent is taken into the cell, binds to the predetermined sequence, and blocks transcription, translation, or S replication thereof. The requirements for selective binding of the antisense aqent are known te.g., a length of 17 bases for selective binding within the human genome).
The present invention is explained in greater detail in the following non-limiting examples.
~..
EZAMP~E I
A. 8vnthesist of 2'-Deox~ribonucleoside-N-Cvanoboranest 8c~eme 1 5 ~8 ~/~B ' H~/~B
Ph3F~H2a~\ / Bu,~F \ /
THF/Renux\J THF/RT \~/
R . Si(CH~CH3)2)3 8 . Gua (a), Ino (b), Ado (c). Cyt (d) or Thy (9) :~S ~ NII ~N~ </~IN~
~ ~ ~ .
Gua-Ino- Ad 9 C y t Cyanoborane adducts of 2'-deoxynucleosides, for example, 2'-deoxyguanosine-N7-cyanoborane t3a), 2'-deoxyinosine-N7-cyanoborane (3b), 2'-deoxyadenosine-Nl-cyanoborane (3c), and 2'-deoxycytidine-N3-cyanoborane (3d), were prepared by an exchange reaction of -- SUBSTITUTE- SHEET
. ;-. .` ~. -.
.
.. ..
2071 9~fi triphenylphosphine-cyanoborane (Ph,PB~{2CN) with 3', 5'-0-protected nucleosides. The 3'- and 5'-~ydroxy groups were protected as silyl ethers by reaction with excess chlorotriisopropylsilane at room temperature in DMF in the presence of imidazole. The exchange was carried out in anhydrous THF at reflux temperature using 2 equiv of Ph3PBH2CN. No increase in the amount of product could be observed (by TLC) after 2-3 hours.
The major products were purified by flash chromatography. Yields can be expected to be approximately 72% for the guanine derivative (2a), 59%
for the adenine derivative (2c), 46% for the cytosine derivative (2d), and 26% for the hypoxanthine derivative (2b). While the first three adducts were readily purified, 2b was obtained only in ca. 95%
purity (by IHNMR). We have as yet been unsuccessful in preparing the boron-substituted thymidine derivative from le by exchange reaction with Ph,PBH2CN.
Deprotection of boronated nucleosides 2a-d with tetra-n-butylammonium fluoride (Bu,NF) to give 3a-d was complete within 0.5 h. Purification was achieved by flash chromatography, followed by crystallization -from MeOH/Et20. Satisfactory (within plus or minus 0.25S) C, ~l, N analyses were obtained for the final compounds. The yields ranged from 44% for 3d to 55%
for 3b.
The site of boron coordination was determined by l5N NMR spectroscopy on a JEOL FX9OQ instrument. No peak was observed for the cooordinated nitrogen (in ~oth coupled and decoupled spectra) due to quadrupole broadening by boron. The absence of peaks for N7 of Gua* and Ino*, Nl of Ade*, and N3 of Cyt* established these to be the sites of ~H2CN coordination. When 15N
NMR spectra of 2a and 2d were obtained on a GE GN500 system, the above quadrupole effect was not observed.
In this case, upfield shifts of-56.6 ppm for N7 of Gua*
and 50.1 ppm for N3 of Cyt* confirmed the assignments.
: .
. ' ~ .
207193fi The shifts upon boronation are qualitatively similar to but lower than those observed upon protonation (ca.65-70 ppm). The coordination site in Gua~ and Ino~ is away from the sites required for Watson-Crick base ~-pairing and should not affect pairing to a large extent. Variable-temperature IH NMR studies on the Gua*-Cyt base pair indeed show the ~ bonding to be approximately as strong as in a normal Gua-Cyt base pair. The coordination at Nl in Ade* and N3 in Cyt~, -o however, should completely disrupt the base pairing, and if incorporated into DNA, these nucleosides may lead to inhibition of replication.
By HPLC, compounds 3a, 3b, and 3d in aqueous medium (0.01 M ~EAAc) are >94% stable over a period of 168 h. Compound 3b, however, is >50% decomposed during this period. The good stability of 3a, 3c, and 3d in aqueous medium makes these compounds suitable for pharmacological testing.
~. Experimental (1). ~, 5~-o-Biq ~triisopropy~silyl)-2~-deoxynucleosides- 2'-Deoxynucleoside [In the case of 2'-deoxycytidine, the hydrochloride was used with an additional equivalent of imidazole] (19.90 mmol) and excess imidazole (96.36 mmol) were taken in anhydrous dimethylformamide (55 ml) under inert atmosphere.
Chlorotriisopropylsilane (51.33 mmol) was added to this mixture and it was stirred at room temperature for 24 ' h. After dilution with diethylether (90 ml), it was washed with a saturated solution of sodium chloride (5 x 80 ml). The organic layer was filtered to remove any suspended solid, dried over anhydrous sodium sulfate and solvent was removed under reduced pressure to give crude product. Purification was achieved by flash chromatography on silica gel using a mixture of solvents (vide infra). The products were characterized :- . . . :
. ~
- , ;
~ n r~
~ . . ~J
by IH nmr, l3C nmr, ~5N nmr, infrared and FAB mass spectroscopic tecllniques.
(la). 3', 5'-0-Bis(triisopropylsilyl)-2'-deoxyguanosine: Purification solvent :
dichloromethane/acetone (6:4); yield = 88.6~; Mp =
decomp above 181-C.;
(lb). 3', 5'-0-Bis(triisopropylsilyl)-2'-deoxyinosine: Purification solvent:
dichloromethane/acetone (6:4); Rf = 0.33; yield =
72.7%; Mp = 158 - 160-C with prior shrinking between 63-sO-c.
(lc). 3', 5'-0-~is(triisoproylsilyl)-2'-deoxyadenosine: Purification solvent :
dichloromethane/acetone (8.5:1.5); Yield = 46.8~; Mp =
130.5 - 131.5-C.
(ld). 3', 5'-Bis-(triisopropylsilyl)-2'-deoxycytidine: Purification solvent :
dichloromethane/acetone (4:6); Rf = 0.33; yield =
83.7%; Mp = 102-C with prior shrinking and color change from white to transparent.
l2). 3'. 5 ~-o-~i9 (triisopropyl)-2'-deoxynucleo~ide- -cyanoborane adaucts: 3', 5'-0-Bis(triisopropylsilyl)-2'-dexoxynucleoside and triphenylphosphine-cyanoborane (2-fold) were taken in anhydrous THF under inert atmosphere and were heated at reflux. After ca 2-3 h, no further change in the ration of product to the starting material could be observed by tlc. The mixture was heated an additional hour, allowed to cool and the solvent was removed under reduced pressure.
The residue was taken in diethyl ether, filtered and the solid was repeatedly washed with diethyl ether.
The filtrate and the washings were concentrated and the crude product was purified by flash chromatography.
The products were characterized by 'H nmr, IlB nmr, ~'C
nmr, "N nmr, infrared and FAB mass spectroscopic techniques.
'- ' ' " ~'., ;'' -' , :' , .
.
, 2 ~ n ^ ,---19-- .
(2a). 3',5'-0-Bis(triisopropylsilyl)-2'-deoxyguanosine N7-cyanoborane: Purification solvent :
dichloromethane/ aceto~e (8:2); yield = 72.1%; Mp =
170-171-C.
(2b). 3', 5'-0-8is(triisopropylislyl)-2'-deoxyinosine-N7-cyanoborane: Purification solvent :
dichloromethane/ acetone (7.5:2.5); Rf = 0.43; yield =
26.2%; contains ca 5% impurity.
(2c). 3', 5'-0-Bis(triisopropylsilyl)-2'-lo deoxyadenosine-Nl-cyanoborane: Purification solvent :
hexane/ethyl acetate (6:4); yield = 58.6%; Mp = 132.5 -133.5, melts with decomposition.
(2d). 3', 5~-0-Bis(triisopropylsilyl)-2~-deoxycytidine-N3-cyanoborane, 2d : Purification solvent : dichloromethane/acetone (9.25:0.75); Rf = 0.40; yield = 45.9%; Mp = 164-165-C.
3. 2'-Deoxynucleoside-cyanoborane adducts:
To a solution of 3', 5'-0-bisttriisopropylsilyl)-2'-deoxynucleoside-cyanoborane adduct (ca 1 g.) in tetrahydrofuran was added tetrabutylammonium fluoride (2 equivalent of 1.1 M solution in tetrahydrofuran). ~-The mixture was stirred at room temperature. After complete reaction (ca 0.5 h), the solvent was removed under reduced pressure. The residual oil was taken in diethyl ether (2 x 50 ml), stirred for a minute, allowed to stand for 5 minutes and then the ether was decanted. The residue was partially purified by flash chromatography on silica gel using dichloromethane/ ;
methanol (8.5:1.5). Fractions containing the desired product were concentrated and the still impure product was finally purified by crystallization from methanol/Et20. The products were characterized by IH
nmr, IIB nmr, ~C nmr, infrared and FAB mass spectroscopic techniques and elemental analysis.
Yields of 45% and better are obtained.
(3a). 2'-Deoxyguanosine-N7-cyanoborane:
yield = 45.5%; Mp = decomp. above 198-C.
~ ~ . . . .
. ~
' :,' ' ':: - ; ' '; : , .
207193~
(3b). 2'-Deoxyinosine-N'-cyanoborane: yield =
54.7~; Mp = decomp. above 173-C.
(3c). 2'-Deoxyadenosine-N1-cyanoborane:
yield = 47.3~; Mp = decomp. a~ove 150-C.
(3d). 2' Deoxycytidine-N3-cyanoborane: yield = 44.3%; Mp = decomp. above 144-C.
EXAMPLE II
Cytotoxic Actlvitv In studies on the antitumor activity, these lo compounds, particularly 3c and 3d, showed potent activity in, among others, the T molt-3 and human colorectal adenocarcinoma screens. More specifically, the cytotoxic activity of the boron adducts of the present invention was tested on the following neoplastic cell lines:
1. L1210 lymphoid leukemia cells, R.
- Geran et al., Cancer Chemotherapy Reports 3, 7 (1972).(grown in RPMI + 15% FBS +
antibiotics).
2. Tmolts acute lymphoblastic T cell leukemia, S. Schreiber and N. Ikemoto, Tett.
Lett 29, 3211 (1988) (grown in RPMI - 1640 +
10% FBS).
3. Colon adenocarcinoma SW480 human colorectal carcinoma. A. Leibovitz et al., Cance~ Res. 36, 4562 (1976) (grown in LlS +
10% FBS).
BORONATED NUCLEOSIDES
Field of the Invention This invention pertains to novel Boron containing compounds having pharmaceutical activity.
More specifically, compounds of the present invention include nucleoside analogues having antineoplastic activity. These compounds seemingly function as antimetabolites with additional utility in Boron Neutron Capture Therapy (BNCT).
Introduction Antimetabolites are a well known class of antineoplastic agents that function by interfering with -nucleic acid sythesis and consequently, replication within the target cells. Some of these compounds structurally mimic biosynthetic precursors of the naturally occurring nucleic acids, which are essential for replication and cell survival. By replacing these precursors, but without exhibiting the same biological effect, these compounds disrupt replication resulting in the demise of the the target cell.
Many antimetabolites have significant antiviral and antitumor activity and are incorporated in a variety of therapeutic regimens. But despite the therapeutic benefits of such compounds, their use is ~' , . ,...',,~,,, ' :' . ',','. -.
:: : . ' : , ' :.
~' :.. ' ' .' . . -2071 '~fi often acco~panied by deleterious side effects, e.g.
nausea, alopecia, and bone marrow depression.
Accordingly, a great deal of in~erest has focused on synthesizing new analogues with improved therapeutic indexes.
We have recently discovered that boron containing nucleotides may be one class of improved nucleic acid analogues. Some exemplary boronated nucleotides are described in copending, commonly owned U.S. patent application Serial No. of B.
Spielvogel, A. Sood, I. Ilall, and B. Ramsay-Shaw titled "Oligoribonucleoside and Oligodeoxyribonucleoside Boranophosphates~' and filed November 30, 1989, which is incorporated herein by reference. There we describe, lS for example, boronated oligonucleotides that contain a boron functionality attached to internucleotide phosphorus.
Boron containing compounds are also useful in an antineoplastic regimen known as Boron Neutron Capture Therapy (BNCT). Soloway, A.H., Prog~ess in :
Boron Chemistry; Steinberg, H., McCloskey, A. L., Eds.;
the Macmillan Company: New York, 1964; Vol. 1, Chapter 4, 203-234. BNCT requires two components (Boron-10 and low energy thermal neutrons) for a radiotoxic reaction.
The inherent advantage is that each component can be manipulated independently to produce the desired radiation effect. Boron-10 has a high cross section for thermal neutrons and after neutron capture, the particles generated, Li & ~, are relatively large by radiation standards and thus have a relatively short track in tissue, 10-14 microns. The Boron-10 is non-radioactive and for use in BNCT, its compounds do not have to be cytotoxic towards tumor cells. Thermal neutrons have such low energy that they cannot ionize tissue components per se. Upon neutron capture, however, the energy generated is sufficient to destroy the cell. The problem in making this procedure :. : :
. .
- - :. . ~ ., , , " , , ~
v clinically effective has stemmed not from the concept, per se, but from lack of knowledge in medicinal chemistry, nuclear engineering and physics, immunology, physiology and pharmacology. The present invention arose from our continued research on new boron-containing compounds having pharmaceutical activity.
8ummary of the Invention A first aspect of the present invention is a boronated nucleoside comprising a nucleoside which is lo N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -~H3, and -BH2COOR, wherein R is Cl to C18 alkyl. Preferably, R is Cl to C9 alkyl, and most preferably R is Cl to C4 alkyl.
A second aspect of the present invention is a boronated nucleotide comprising a 5' phosphate ester of a boronated nucleoside as described above.
A third aspect of the present invention is a boronated oligonucleotide comprising a chain of natural or modified ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a boronated nucleotide as described above.
Nucleosides of the present invention have phar~aceutical activity, including antihyperlipidemic, antiinflammatory, and antineoplastic activity.
Nucleotides of the present invention are useful as intermediates for making oligonucleotides of the present invention. Oligonucleotides of the present .
invention are useful as antisense agents and probes.
A method for synthesizing N-boronated nucleosides of the present invention from a substrate nucleoside, which substrate nucleoside is comprised of a ribose moiety covalently bound to a nitrogenous base, is also disclosed. The method comprises:
(a) protecting the hydroxy substituents of the ribose; then , :. ,. . - : .:
: ~
~n7~
(b) boronating the nitrogenous base; and then (c) deprotecting the hydroxy substituents.
Preferably, the nitrogenous base is boronated by 5 reacting the substrate nucleoside with either polymeric BH2CN or LX, wherein L is a Lewis base and X is a boron-containing substituent selected from the group consisting of -BH2CN, -BH" and -B)~2COOR, wherein R is as given above.
The term alkyl, as used herein, refers to linear or branched alkyl groups which may be saturated or unsaturated. Preferred alkyl groups are linear and saturated.
Detailed Description of the Invention We have successfully synthesized novel boronated nucleosides that have surprising antineoplastic, antiinflammatory and antihyperlipidemic properties. The present invention provides novel pharmaceutical agents, methods for their synthesis, methods for treating patients in need of such treatment, and pharmaceutical formulations comprising such agents.
The nucleosides of the present invention comprise nucleosides having a boron-containing substituent on the base. More specifically, the present invention provides N-boronated nucleosides wherein the boronated nitrogen is a ring nitrogen of the base covalently bonded to the boron of the boron-containing substituent. The base is a purine or a pyrimidine or an analog thereoE (as discussed in detail below). A preferred group of bases are those selected from the group consisting of adenine, cytosine, guanine, inosine, and the analogs thereof.
By nucleosides we mean a purine or pyrinidine base, and analogs thereof, linked to a pentose. The pentose is preferably D-ribose, 2'-deoxy-D-ribose or 2',3'-dideoxy-D-ribose. Thus the nucleosides of the . ' ' ' ' , . ' . . .
~ n ~
~ U . _ L~ J
present invention include both naturally occurrin~
nucleosides and analogs thereof.
In the case of a purine nucleoside, boronation occurs preferentially at either the Nl or N7 position depending upon the purine. For example, when the purine is adenine, boronation occurs at the Nl position. In the case of guanine, boronation occurs at the N7 position. It is contemplated that boronation at other nitrogens on the nitrogenous bases will yield lo additional useful agents.
The nucleoside base may generally be a natural base, such as adenine, thymine, cytosine, guanine, uracil, xanthine, or hypoxanthine, (the latter two being the natural degradation products) or an analog thereof as found in, for example, 4-acetylcytidine; 5-(carboxyhydroxyl- methyl)uridine; 2'-0-methylcytidine; 5-carboxymethyl- aminomethyl-2-thiouridine; 5-carboxymethylaminomethyl-uridine;
dihydrouridine; 2'-0-methylpseudo-uridine; beta,D-galactosylqueosine; 2'-0-methylguanosine; N6-isopentenyladenosine; l-methyladenosine; l-methyl-pseudo-uridine; l-methylguanosine; l-methylinosine;
2,2-dimethylguanosine; 2-methyladenosine; 2-methyl-guanosine; 3-methyl-cytidine; S-methylcytidine; N6-methyladenosine; 7-methyl-guanosine; 5-methylamino-methyluridine; 5-methoxyamino-methyl-2-thiouridine;
beta,D-mannosylqueosine; 5-methoxy-carbonylmethyl-uridine; 5-methoxyuridine; 2-methylthio-N6-isopentenyl-adenosine; N-((9-beta-D-ribofuranosyl-2-methylthio-purine-6-yl)carbamoyl)threonine; N-((9-beta-D-ribofuranosyl- purine-6-yl)N-methyl-carbamoyl)-threonine; uridine-5-oxyacetic acid methylester;
uridine-5-oxyacetic acid (v); pseudouridine; queosine;
2-thiocytidine; 5-methyl-2-thiouridine; 2-thiouridine;
4-thiouridine; 5-methyluridine; 2'-O-methyl-5-methyluridine; and 2'-O-methyluridine. Preferred analogs are the methylated analogs.
.
.~ .
~o J ~u~J
Exemplary boronated nucleosides of the present invention are:
(A) ~uanosine-N7-cyanoborane;
(B) inosine-N7-cyanoborane;
(C) adenosine-Nl-cyanoborane;
(D) cytidine-N3-cyanoborane;
(E) guanosine-N7-borane;
(F) inosine-N7-borane;
(G) adenosine-Nl-borane; .-:
(T~) cytidine-N3-borane;
(I) guanosine-N7-carbomethoxyborane;
(J) inosine-N7-carboethoxyborane;
(K) adenosine-Nl-carbopropoxyborane;
(L) cytidine-N3-carbobutoxyborane;
(M) 2'-deoxyguanosine-N7-cyanoborane;
(N) 2'-deoxyinosine-N7-cyanoborane;
(0) 2'-deoxyadenosine-Nl-cyanoborane;
(P) 2'-deoxycytidine-N3-cyanoborane;
(Q) 2'-deoxyguanosine-N7-borane;
(R) 2'-deoxyinosine-N7-borane;
(S) 2'-deoxyadenosine-Nl-borane;
~T) 2'-deoxycytidine-N3-borane;
(U) 2'-deoxyguanosine-N7-carbomethoxyborane;
(V) 2'-deoxyinosine-N7-carboethoxyborane;
(W) 2'-deoxyadenosine-N1-carbopropoxyborane;
(X) 2'-deoxycytidine-N3-carbobutoxyborane; ~ .
(Y) 2',3'-dideoxyguanosine-N7-cyanoborane;
(Z) 2',3'-dideoxyinosine-N7-cyanoborane;
(AA) 2',3'-dideoxyadenosine-Nl-cyanoborane;
(A3) 2',3'-dideoxycytidine-N3-cyanoborane;
(AC) 2',3'-dideoxyguanosine-N7-borane;
(AD) 2',3'-dideoxyinosine-N7-borane;
(AE) 2',3'-dideoxyadenosine-Nl-borane;
(AF) 2',3'-dideoxycytidine-N3-borane; -(AG) 2',3'-dideoxyguanosine-N7-carbomethoxyborane;
.
.:,. - - : . :
: , - ',: . .~ ~ ' - ' : , , ' ' :, . .
.
207~9~fi (AH) 2',3'-dideoxyinosine-N7-carboethoxyborane;
tAI) 2',3'-dideoxyadenosine-~11-carbopropoxyborane; and S tAJ) 2~3~-dideoxycytidine-N3 carbobutoxyborane.
Particularly preferred boronated nucleosides are: 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2'-deoxyadenosine-Nl-cyanoborane, and lo 2'-deoxycytidine-N3-cyanoborane.
The nucleosides of the present invention further comprise 5'-phosphate esters of the N-boronated nucleosides described herein, such phosphate esters are also known as nucleotides. Such nucleotides, particularly the monophosphates, are protected in conventional manner and used for the synthesis of oligonucleotides, as discussed below. Such phosphate esters include 5' mono-, di- and triphosphates, which may be protected as esters. Additionally, molecules and macromolecules comprising multimers of two or more nucleosides, which may be linked via a 3'-5' phosphate ester, e.g. oligonucleotides (the terms "oligonucleotides~ and "polynucleotides~ being used interchangeably herein), and comprising one or more N-boronated nucleosides are also the subject of the present invention. Accordingly, N-boronated nucleotides, oligonucleotides, and polynucleotides may be used as therapeutic agents and otherwise useful reagents, e.g. diagnostic reagents.
Oligonucleotides of the present invention can be synthesized in accordance with methods that are well known in the art. Such methods include the phosphite method and the phosphotriester method. 1 Chemistry of Nucleosides and Nucleotides, 294ff (L. Townsend ed.
1988). ~he length of the oligonucleotide is not :i critical, as modern synthetic techniques and splicing techniques have made synthetic oligonucleotides of :. . ' .,;, . : , , .
~ u J ~
considerable length feasible. Thus, the oligonucleotide may for example be 2 to 3 nucleotides long, 2 to 18 nucleotides long, 2 to 30 nucleotides long, 2 to 50 nucleotides long, or 50 or more S nucleotides long.
Oligonucleotides containing N-boronated bases may alternatively be prepared, with boronation occur~ng randomly, in essentially the same manner as the nucleoside, but with an oligonucleotide substituted for lo the nucleoside. For example, in such a reaction, the 3' terminus of the oligonucleotides may be immobilized to a solid support (e.g., controlled pore glass), the 5' terminus protected as the dimethyltrityl ether, and amino groups on bases protected with isobutyryl groups.
Oligonucleotides used for such a reaction preferably contain at least one base which is not thymidine or a thymidine analog, as these are less preferred bases for boronation.
Derivatives of the oligonucleotides and polynucleotides may additionally be formed by modifying the internucleotide phosphodiester linkage.
Internucleotide phosphodiester linkages in the chain are modified, for example, to the methylphosphonate, the phosphotriester, the phosphorothioate, the phosphorodithioate, and the phosphoramidate, all as is known in the art.
Additional synthetic analogues of the nucleosides, nucleotides, and oligonucleotides of the present invention may be formed by otherwise modifying the 3' or 5' end of the nucleoside, and any 2' hydroxyl groups. Groups that can be added to the 3' or 5' end vary widely, from relatively inert protecting groups to 1~^
reactive groups or groups with special properties to obtain desireable physical, chemical, or biochemical effects.
A wide variety of protecting groups can be substituted on the 2', 3', and 5' hydroxy groups, such .
!
207193~
as the triphenylmethyl (trityl) group and its derivatives on the 5' hydroxy group, and acetyl, ~enzo~l, or the 3'-O-succinyl group on the 3' hydroxy gro~p, as is known in the art. See 1 Chemistry of rJucleosides and Nucleotides, 287-92 (L. Townsend ed.
1988). In general, the 5' hydroxy group is protected with an acid labile group and the 3' hydroxy group is protected with an acyl group. Id. at 289 (When the 5' hydroxyl group is protected with an acid labile group lo such as mono- and dimethoxytrityl, the 3'-hydroxyl group of deoxynucleosides can be protected with acyl groups.). ~n general, a 2' hydroxy group is protected as a methyl ether, protected with a silyl group, or the 2' and 3' hydroxy groups may be protected together as an acetal.
Reactive groups or groups with special properties may also be attached at the 3' or 5' position. For example, analogs may be formed by joining an intercalating agent to oligonucleotides and polynucleotides in the manner described in U. S. Patent No. 4,835,263 to Nguyen et al. (the disclosure of this and all other patent references cited herein is incorporated herein by reference).
The present invention also provides methods for preparing the compounds of the present invention.
The boronation of a nucleoside is accomplished by first preparing a hydroxy- protected nucleoside. Protecting groups and methods for their use are well known in the art. See, Carey and Sundberg, Advanced Orqanic Chem i try. Part B, pp. 408-414 (1980). Preferred protecting groups are organosilanes, e.g., chlorotriisopropyl silane will form the triisopropyl silyl ether.
As an example, the nucleoside is 0-protected by forming a silyl ether by reaction with excess chlorotriisopropyl-silane at room temperature in the presence of imidazole. The exchange reaction is ' ' ' ' ' `' : .. , ::
effected by reacting the O-protected nucleoside with an organoborane. The organoborane is generally either polvmeric B~2C~ or a compound LX, wherein L is a Lewis base and X is a boron-containing su~stituent as given above. Suitable Lewis bases include amine, phosphine, sulfide, and ether (e.g., tetrahydrofuran). The strength and steric properties of the Lewis base should be chosen so as to provide a suitable leaving group.
Exemplary organoboranes include aniline-cyanoborane, 1~ triphenylphosphine-carboalkoxyboranes (wherein the alkoxy group alkyl is R as given above), dimethylsulfide-borane, and tetrahydrofuran-borane. A
preferred orqanoborane is triphenylphosphine-cyanoborane, in which case the resulting product is an O-protected ribonucleoside N-cyanoborane. The exchange reaction is effected by reaction in anhydrous tetrahydrofuran (THF) at reflux temperature using two ~ -equivalents of triphenylphosphine-cyanoborane. The reaction is at equilibrium in two to three hours.
The resulting O-protected N-boronated ribonucleoside is deprotected by appropriate methods -known in the art. For example, when protected by formation of a silyl ether, deprotection can be effected under hydrolytic or nucleophilic conditions.
In our preferred case the silyl ether is effectively removed under hydrolytic conditions in the presence of fluoride ion, e.g., tetra-n-butylammonium fluoride.
The compounds of the present invention have pharmaceutical activity and are useful in treating mammals (e.g., human, cat, dog, cow, horse, mouse) suffering from one or more of several maladies. These compounds show pharmaceutical activity in killing cancer cells in_vitro, and may be useful in combatting corresponding tumors in vivo. For example, the compounds of the present invention show cytotoxic activity against colorectal carcinoma, adenocarcinoma, osteosarcoma, breast carcinoma and glioma.
.. .
. .
Accordingly, the compounds of the present invention provide a method for treating a tumor bearing mammal comprising administering to said mammal a therapeutically effective amount of a boronated S nucleoside of the present invention. Furthermore, it is contemplated that the antineoplastic efficacy of these compounds can be improved or supplemented by the conjoint administration with other known antineoplastic agents, as, for example, in a combination chemotherapy lo regimen. Exemplary of such other known antineoplastic agents are: vinca alXaloids such as vincristine, vinblastine, and vindesine; epipodophyllotoxins such as etoposide and teniposide; anthracycline antibiotics such as daunorubicin, doxorubicin, mitoxantraone, and bisanthrene; actinomyGin D; and plicamycin.
In addition to the direct inhibition of tumor growth, the preferential localization of boron compounds in the neoplasm of tumor cells will allow the use of boron-10 neutron capture therapy (BNTC) for the ~ -destruction of tumor cells. Moreover, the dual effect of this therapeutic regimen may lower the therapeutically effective amounts of the pharmaceutically active agents, and thereby reduce the ~-deleterious side effects that often accompany the use of such agents. Thus, the present invention also provides methods for treating tumor-bearing mammals in which the mammal is administered a boronated nucleoside as described herein and then exposed to thermal neutron radiation. The thermal neutron radiation is administered in an amount and in a manner effective for 10B located in a tumor by virtue of the administration of the compound of the present invention to the subject to capture a neutron, decay, and release an alpha particle in cells of the tumor.
The compounds of the present invention also have pharmaceutical activity as antiinflammatory agents in mammals. The compounds of the present invention : .
~v ,i~
provide a method for treating a mammal suffering from inflammation comprising administering to said mammal a therapeutically ef~ective amount of an N-boronated nucleoside. The compounds o~ the present invention may provide additional utility when conjointly administered with other known antiinflammatory agents or pain killers or some such pharmaceutical. Exemplary of other known antiinflammatory agents are acetylsalicylic acid (asprin), salicylic acid, diflunisal, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine, dipyrone, apazone, acetaminophen, indomethacin, sulindac,meclofenamate, tolmetin, zomepirac, ibuprofen, and piroxicam.
The compounds of the present invention are useful as hypolipidemic agents. The compounds of the present invention provide a method for treating a mammal suffering from hyperlipidemia comprising administering to said mammal a therapeutically effective amount of an N-boronated nucleoside.
Additionally, the compounds of the present invention provide a method for treating a mammal suffering from hypercholesterolemia comprising administering to said mammal a therapeutically effective amount of an N- -boronated nucleoside. ~y administering these compounds to hyperlipidemic patients the total lipoprotein level may be reduced or the lipoprotein profile may be improved. Furthermore, these compounds may be con~ointly administered with other known hypolipedemic agents to enhance or supplement their efficacy.
Exemplary of such other ~nown hypolipidemic agents are nicotinic acid, clofibrate, gemfibrozil, probucol, cholestyramine, colestipol, compactin, mevinolin, choloxin, neomycin, and beta-sitosterol.
The compounds of the present invention may be administered in any of the variety of modes currently employed for analogous pharmaceutical agents, which modes are well known in the art. For example, these .
` ~
.
~ n ~
compounds may be administered systemically. Systemic administration includes parenteral administration and gastro-enteral administration.
When prepared in a pharmaceutical formulation for parenteral administration the compounds of the present invention should be prepared in a pharmaceutically acceptable carrier such as substantially non-pyrogenic, sterile, parenterally acceptable, aqueous solutions.
Alternatively, the compounds of the present invention may be formulated in pharmaceutical preparations for gastro-enteral administration. Such pharmaceutical preparations include tablets, capsules and suppositories. When formulated for administration according to any of the above methods the pharmaceutical preparations may further comprise buffers, binders, and other pharmaceutically acceptable excipients as are well known in the art.
A therapeutically effective amount of a boronated nucleoside is in the range of about 0.1-100 mg/kg/day. The preferred range is about 0.5-50 mg/kg/day. More preferred is an amount in the range of about 1-10 mg/kg/day. When administered conjointly with other pharmaceutically active agents even less of the boronated nucleoside may be therapeutically effective.
The oligonucleotides of the present invention may be used as probes in a variety of diagnostic techniques. One such diagnostic technique is Magnetic Resonance Imaging (MRI). MRI is a noninvasive technique used to detect the presence and location of tumors in a patient. For example, as contemplated in the present context, cancer cell specific boronated compounds are administered to a patient, whereupon they concentrate in cancerous tissue. The MRI instrument is capable of detecting and locating regions of abnormal concentrations of Boron. By indicating the regions .. . . . . ...
: . :.......... . ~
; . ~
2'~ n~
~J ~ ., V .J
havinq high relative concentrations of Boron, MRI
establ ishes the presence and location of tumors.
Another diagnostic application of the oligonucleotides of the present invention is their use as molecular probes. By incorporating N-boronated nucleosides, or their 5'-phosphate esters, into an oligonucleotide, either at an interior or terminal position, a detectable oligonucleotide probe is constructed that can be used to detect the presence of complementary sequences of DNA or RNA in a sample.
These probes can be used in any suitable environment, such as Southern blotting and Northern blotting, the details of which are known. See e.g., R. Old and S. Primrose, Principles of Gene Manipulation, 8-10 (3d Ed. 1985). When used as probes, the boron atom serves as a radiolabel, though it is not itself radioactive until exposed to thermal neutron radiation (low energy neutrons). When exposed to low energy neutrons, IB absorbs a neutron and forms IlB, which rapidly decays and releases an alpha particle, thus providing a detectable signal. The techniques involved in the generation of the alpha particle are known. See. e.~., A. Soloway, Borax Rev. 3, 7-9 (1988).
Oligonucleotides of the present invention which are capable of binding to polyribonucleic acid or polydeoxyribonucleic acid are useful as antisense agents in the same manner as conventional antisense agents. See gene~allv Antisense Molecular Biology and S-oligos, Syntkesis 1 (Oct. 1988) (published by Synthecell Corp., Rockville, MD); 2 Discoveries in Antisense Nucleic Acids (C. Brakel and R. Fraley eds.
1989). Antisense agents of the present invention may be used by contacting an antisense agent which is capable of selectively binding to a predetermined polydeoxyribonucleic acid sequence or polyribonucleic acid sequence to a cell containing such sequence (e.g., .
. . . . .
.: . . -; . , . .
: . . ., ~.: . ;
.. . : . ~ -. : .
' ::
:. , :.. :: . ...
9 0 / ~ 7 4 4 6 -15- lP ~ Us 0 6 JAN 1992 by adding the antisense agent to a culture medium containing the cell) so that the antisense agent is taken into the cell, binds to the predetermined sequence, and blocks transcription, translation, or S replication thereof. The requirements for selective binding of the antisense aqent are known te.g., a length of 17 bases for selective binding within the human genome).
The present invention is explained in greater detail in the following non-limiting examples.
~..
EZAMP~E I
A. 8vnthesist of 2'-Deox~ribonucleoside-N-Cvanoboranest 8c~eme 1 5 ~8 ~/~B ' H~/~B
Ph3F~H2a~\ / Bu,~F \ /
THF/Renux\J THF/RT \~/
R . Si(CH~CH3)2)3 8 . Gua (a), Ino (b), Ado (c). Cyt (d) or Thy (9) :~S ~ NII ~N~ </~IN~
~ ~ ~ .
Gua-Ino- Ad 9 C y t Cyanoborane adducts of 2'-deoxynucleosides, for example, 2'-deoxyguanosine-N7-cyanoborane t3a), 2'-deoxyinosine-N7-cyanoborane (3b), 2'-deoxyadenosine-Nl-cyanoborane (3c), and 2'-deoxycytidine-N3-cyanoborane (3d), were prepared by an exchange reaction of -- SUBSTITUTE- SHEET
. ;-. .` ~. -.
.
.. ..
2071 9~fi triphenylphosphine-cyanoborane (Ph,PB~{2CN) with 3', 5'-0-protected nucleosides. The 3'- and 5'-~ydroxy groups were protected as silyl ethers by reaction with excess chlorotriisopropylsilane at room temperature in DMF in the presence of imidazole. The exchange was carried out in anhydrous THF at reflux temperature using 2 equiv of Ph3PBH2CN. No increase in the amount of product could be observed (by TLC) after 2-3 hours.
The major products were purified by flash chromatography. Yields can be expected to be approximately 72% for the guanine derivative (2a), 59%
for the adenine derivative (2c), 46% for the cytosine derivative (2d), and 26% for the hypoxanthine derivative (2b). While the first three adducts were readily purified, 2b was obtained only in ca. 95%
purity (by IHNMR). We have as yet been unsuccessful in preparing the boron-substituted thymidine derivative from le by exchange reaction with Ph,PBH2CN.
Deprotection of boronated nucleosides 2a-d with tetra-n-butylammonium fluoride (Bu,NF) to give 3a-d was complete within 0.5 h. Purification was achieved by flash chromatography, followed by crystallization -from MeOH/Et20. Satisfactory (within plus or minus 0.25S) C, ~l, N analyses were obtained for the final compounds. The yields ranged from 44% for 3d to 55%
for 3b.
The site of boron coordination was determined by l5N NMR spectroscopy on a JEOL FX9OQ instrument. No peak was observed for the cooordinated nitrogen (in ~oth coupled and decoupled spectra) due to quadrupole broadening by boron. The absence of peaks for N7 of Gua* and Ino*, Nl of Ade*, and N3 of Cyt* established these to be the sites of ~H2CN coordination. When 15N
NMR spectra of 2a and 2d were obtained on a GE GN500 system, the above quadrupole effect was not observed.
In this case, upfield shifts of-56.6 ppm for N7 of Gua*
and 50.1 ppm for N3 of Cyt* confirmed the assignments.
: .
. ' ~ .
207193fi The shifts upon boronation are qualitatively similar to but lower than those observed upon protonation (ca.65-70 ppm). The coordination site in Gua~ and Ino~ is away from the sites required for Watson-Crick base ~-pairing and should not affect pairing to a large extent. Variable-temperature IH NMR studies on the Gua*-Cyt base pair indeed show the ~ bonding to be approximately as strong as in a normal Gua-Cyt base pair. The coordination at Nl in Ade* and N3 in Cyt~, -o however, should completely disrupt the base pairing, and if incorporated into DNA, these nucleosides may lead to inhibition of replication.
By HPLC, compounds 3a, 3b, and 3d in aqueous medium (0.01 M ~EAAc) are >94% stable over a period of 168 h. Compound 3b, however, is >50% decomposed during this period. The good stability of 3a, 3c, and 3d in aqueous medium makes these compounds suitable for pharmacological testing.
~. Experimental (1). ~, 5~-o-Biq ~triisopropy~silyl)-2~-deoxynucleosides- 2'-Deoxynucleoside [In the case of 2'-deoxycytidine, the hydrochloride was used with an additional equivalent of imidazole] (19.90 mmol) and excess imidazole (96.36 mmol) were taken in anhydrous dimethylformamide (55 ml) under inert atmosphere.
Chlorotriisopropylsilane (51.33 mmol) was added to this mixture and it was stirred at room temperature for 24 ' h. After dilution with diethylether (90 ml), it was washed with a saturated solution of sodium chloride (5 x 80 ml). The organic layer was filtered to remove any suspended solid, dried over anhydrous sodium sulfate and solvent was removed under reduced pressure to give crude product. Purification was achieved by flash chromatography on silica gel using a mixture of solvents (vide infra). The products were characterized :- . . . :
. ~
- , ;
~ n r~
~ . . ~J
by IH nmr, l3C nmr, ~5N nmr, infrared and FAB mass spectroscopic tecllniques.
(la). 3', 5'-0-Bis(triisopropylsilyl)-2'-deoxyguanosine: Purification solvent :
dichloromethane/acetone (6:4); yield = 88.6~; Mp =
decomp above 181-C.;
(lb). 3', 5'-0-Bis(triisopropylsilyl)-2'-deoxyinosine: Purification solvent:
dichloromethane/acetone (6:4); Rf = 0.33; yield =
72.7%; Mp = 158 - 160-C with prior shrinking between 63-sO-c.
(lc). 3', 5'-0-~is(triisoproylsilyl)-2'-deoxyadenosine: Purification solvent :
dichloromethane/acetone (8.5:1.5); Yield = 46.8~; Mp =
130.5 - 131.5-C.
(ld). 3', 5'-Bis-(triisopropylsilyl)-2'-deoxycytidine: Purification solvent :
dichloromethane/acetone (4:6); Rf = 0.33; yield =
83.7%; Mp = 102-C with prior shrinking and color change from white to transparent.
l2). 3'. 5 ~-o-~i9 (triisopropyl)-2'-deoxynucleo~ide- -cyanoborane adaucts: 3', 5'-0-Bis(triisopropylsilyl)-2'-dexoxynucleoside and triphenylphosphine-cyanoborane (2-fold) were taken in anhydrous THF under inert atmosphere and were heated at reflux. After ca 2-3 h, no further change in the ration of product to the starting material could be observed by tlc. The mixture was heated an additional hour, allowed to cool and the solvent was removed under reduced pressure.
The residue was taken in diethyl ether, filtered and the solid was repeatedly washed with diethyl ether.
The filtrate and the washings were concentrated and the crude product was purified by flash chromatography.
The products were characterized by 'H nmr, IlB nmr, ~'C
nmr, "N nmr, infrared and FAB mass spectroscopic techniques.
'- ' ' " ~'., ;'' -' , :' , .
.
, 2 ~ n ^ ,---19-- .
(2a). 3',5'-0-Bis(triisopropylsilyl)-2'-deoxyguanosine N7-cyanoborane: Purification solvent :
dichloromethane/ aceto~e (8:2); yield = 72.1%; Mp =
170-171-C.
(2b). 3', 5'-0-8is(triisopropylislyl)-2'-deoxyinosine-N7-cyanoborane: Purification solvent :
dichloromethane/ acetone (7.5:2.5); Rf = 0.43; yield =
26.2%; contains ca 5% impurity.
(2c). 3', 5'-0-Bis(triisopropylsilyl)-2'-lo deoxyadenosine-Nl-cyanoborane: Purification solvent :
hexane/ethyl acetate (6:4); yield = 58.6%; Mp = 132.5 -133.5, melts with decomposition.
(2d). 3', 5~-0-Bis(triisopropylsilyl)-2~-deoxycytidine-N3-cyanoborane, 2d : Purification solvent : dichloromethane/acetone (9.25:0.75); Rf = 0.40; yield = 45.9%; Mp = 164-165-C.
3. 2'-Deoxynucleoside-cyanoborane adducts:
To a solution of 3', 5'-0-bisttriisopropylsilyl)-2'-deoxynucleoside-cyanoborane adduct (ca 1 g.) in tetrahydrofuran was added tetrabutylammonium fluoride (2 equivalent of 1.1 M solution in tetrahydrofuran). ~-The mixture was stirred at room temperature. After complete reaction (ca 0.5 h), the solvent was removed under reduced pressure. The residual oil was taken in diethyl ether (2 x 50 ml), stirred for a minute, allowed to stand for 5 minutes and then the ether was decanted. The residue was partially purified by flash chromatography on silica gel using dichloromethane/ ;
methanol (8.5:1.5). Fractions containing the desired product were concentrated and the still impure product was finally purified by crystallization from methanol/Et20. The products were characterized by IH
nmr, IIB nmr, ~C nmr, infrared and FAB mass spectroscopic techniques and elemental analysis.
Yields of 45% and better are obtained.
(3a). 2'-Deoxyguanosine-N7-cyanoborane:
yield = 45.5%; Mp = decomp. above 198-C.
~ ~ . . . .
. ~
' :,' ' ':: - ; ' '; : , .
207193~
(3b). 2'-Deoxyinosine-N'-cyanoborane: yield =
54.7~; Mp = decomp. above 173-C.
(3c). 2'-Deoxyadenosine-N1-cyanoborane:
yield = 47.3~; Mp = decomp. a~ove 150-C.
(3d). 2' Deoxycytidine-N3-cyanoborane: yield = 44.3%; Mp = decomp. above 144-C.
EXAMPLE II
Cytotoxic Actlvitv In studies on the antitumor activity, these lo compounds, particularly 3c and 3d, showed potent activity in, among others, the T molt-3 and human colorectal adenocarcinoma screens. More specifically, the cytotoxic activity of the boron adducts of the present invention was tested on the following neoplastic cell lines:
1. L1210 lymphoid leukemia cells, R.
- Geran et al., Cancer Chemotherapy Reports 3, 7 (1972).(grown in RPMI + 15% FBS +
antibiotics).
2. Tmolts acute lymphoblastic T cell leukemia, S. Schreiber and N. Ikemoto, Tett.
Lett 29, 3211 (1988) (grown in RPMI - 1640 +
10% FBS).
3. Colon adenocarcinoma SW480 human colorectal carcinoma. A. Leibovitz et al., Cance~ Res. 36, 4562 (1976) (grown in LlS +
10% FBS).
4. Lung bronchogenic MB-9812, S.
Aronson et al., Expt. Cell Res. 61, 1 (1970) (grown in EMEM + 10% F8S + NEAA).
Aronson et al., Expt. Cell Res. 61, 1 (1970) (grown in EMEM + 10% F8S + NEAA).
5. Human Osteosarcoma TE418. H. Smith et al., Int J._Cancer 17, 219 (1976) (grown in DMEM + 10% FBS).
6. KB epidermoid oral nesopharnyx. R.
Geran, su~ra; H. Eagle, Proc. Soc. Expt.
, ' 207193~
Biol~ 89, 362 (1955)(grown in EMEM + 5~ calf serum).
Geran, su~ra; H. Eagle, Proc. Soc. Expt.
, ' 207193~
Biol~ 89, 362 (1955)(grown in EMEM + 5~ calf serum).
7. ~lela-S, A~CC-CCL 2.2, cervical carcinoma suspended, S. Schreiber and N.
I~emoto, su~ra; T. Puck et al., J. Exp. Med.
103, 273 (1956) (grown in Joklik + 5% FBS, Ham's Fl2 + 5~ FBS).
I~emoto, su~ra; T. Puck et al., J. Exp. Med.
103, 273 (1956) (grown in Joklik + 5% FBS, Ham's Fl2 + 5~ FBS).
8. Breast carcinoma MDA MB157,W.
Nelson-~ees et al., Int J. Cancer 16, 74 (1975) (grown in EMEM + 10% FBS + NEAA).
Nelson-~ees et al., Int J. Cancer 16, 74 (1975) (grown in EMEM + 10% FBS + NEAA).
9. ~uman glioma cell EH 118 MG
transformed stain of Rous sarcoma virus, J.
Lutton et al., J. Biol. Chem. 254, 11181 (lS79) (grown in DMEM-H + 10% FCS).
The cytotoxic screens were conducted ~-according to NIH protocols, see E. Huang et al., J.
Pharm. Sci. 61, 108 (1972), with 10' cells, growth medium, antibiotics and drugs from 0.5 to 100 ~g/ml final concentration. For the L1210, l~ela-S, amd Tmolt, (i.e. the suspended cells), the incubations were conducted in sterile test tubes in a final volume of 1 ml for 72 hr at 37 C in a CO2 incubator. The cells on -the third day were still in log growth phase. The number of cells/ml are determined using trypan blue exclusion and a hemocytometer. See. e.g., R. Geran, sup~. For solid tumors there is plated out in wells 1 x 10' cells with 1 ml of medium + antibiotics and the other components of growth. When the controls have converged (~95%) then the density of the cells is estimated and the ED,o values calculated. These structures are given in Table I below, and these data are given in Table II below.
. . .
TABLE I
8tructure of_Compounds Deox~ribose Nucleoside 8ase 3' 5' -I - 7 3b Ino OH OH H BH2CN
2b InoSi(CH(CH,)2)3 Si(CH(CH,)2), 1{ 8H2CN
3a Gua OH OH ~{ BH2CN
102a GuaSi(CH(CH3) 2) ~ Si (cH(CH3) 2) 3 H BH2CN
3c Ade OH OH BH2CN H
2c AdeSi(CH(CH3) 2) 3 Si(~(C}I,) 2) 3 BH2CN 1l N~
3d Cyt OH OH BH2CN --152d CytSi(CH(CH3)2)3 Si(CH(CH3)2)3 BH2CN __ 4. CytH Si(CH(CH3) 2 ) 3 BH2CN - -TABLE II
Cytotoxicity of Boron Adductq of ribonucleosides Adeno Lung Carci- Bron- Osteo noma cho- sar-C~d. L1210 P388 Tmolt SW480 KB qenic Hela-S coma Glioma 3b 4.00 7.21 1.73 3.86 3.45 4.93 2.41 7.28 8.49 2b 4.00 7.31 6.49 2.57 5.29 6.20 3.26 NA 6.21 3a 5.08 5.87 2a 6.05 5.13 2.26 2.67 2.67 4.05 2.83 7.20 7.61 3c 3.68 5.96 1.36 2.82 3.12 5.26 2.35 7.39 6.88 2c 6.86 7.46 1.88 2.87 5.79 5.58 3.42 NA 4.99 3d 2.61 3.17 1.13 2.96 5.46 3.73 3.37 4.63 4.39 --2d 4.78 7.26 3.36 2.73 5.72 6.60 2.94 NA 6.77 4. 2.92 -- 2.34 2.71 4.07 5.58 3.63 6.62 3.~3 2n7~ 9~
EXAMPLE IlI
AntihyperliDidemic Activitv Compounds to be tested were suspended in 1%
aqueous carboxymethylcellulose, homogenized and administered to male CF~ mice (25 g) intraperitoneally for 16d. On days 9 and 16, blood was obtained by tail vein bleeding, and the serum was separated by centrifugation for three minutes. The serum cholesterol levels were determined by a modification of the Liebermann-Burchard reaction in accordance with known techniques. See A. Ness, et al., Clin. Chim.
Acta 10, 229 (1964). Serum triglyceride levels were determined with a commercial kit, the Fisher llycel Triglyceride Test Kit, for a different group of animals bled on day 16. The results of these antihyperlipidemic screens, for a compound dose of 8 mg/kg, are shown in Table III below.
EXAMPLE IV
Anti-Inflammatory Activity CF, male mice (-25 g) were administered test drugs at 5-40 mg/kg in 0.05% Tween 80-H20 intraperitoneally 3 hours prior to and 30 minutes prior to the injection of 0.2 ml of 1% carraqeenan in 0.9%
saline into the plantar surface of the right hind foot.
Saline was injected into the left hind foot, which serves as a base line. After 3 hours, both feet were excised at the tibiotarsal (ankle) joint according to standard procedures. See C. Winter et al., Proc. Soc.
Expt. Biol Med. 544, 111 (1962); A. Roszkawski et al., J._Pharm. Exp. Ther. 179, 114 (1971). Control mice afforded a 78+3 mg increase in paw weiqht. Data on the percent inhibition of the inflammatory response for a dose of 8 mg/kg are reported in Table III below.
.
. . . .. ..
'' , : ' . - ~ ' , .~
. ' . ~ . , .
.~; :' ' TA~LE III
Antiinflammatory and }Iypolipidemic ~ctivities of ~oron-Containinq Nucleo~3ide~3 Antiinflammatory llvpolipidemic Cholesterol Triglyceride Activity at Inhibition at Inhibition at 8 mg/kg/day 8 mg/kg/day 8 mg/kg/day Compo~lnd ~ inhibition % inhibition % inhibitlon 3a 22 44 37 lo 3b 25 19 30 3c 22 44 37 3d 52 38 23 Standard 47 (phenyl- 13 (clofib- 2S
butazone) rate at 130 mg/
~g/day)n EXAMPLE V
A. Synthesis of oligomers ~cheme II
G' G^^
20DMT--o~o ¦ DMr~)~l~o I DI~T-O ~0 H\f~H m l ~ 201- Hr\~ H ~ nDu ~ NE' H I~ ir~3Sio H lPr]Sl IllU
G^^ G'' DI~T - C~ ~¦ C l P I O~le l N I r ~ 2 ~ ~¦ . -H\ j~/H l~\~H
HO HMcO - I ~ ~ 11 r 1 ~ lll 2C~
IcH3)2cllc~-C~N~ (Cll3) 2CIICN - l T ~
Step 1 Preparation of 5'-DMT-3'-TIPSI-N2-isobutyryl-2'-deoxyguanosine.
: . . . . . ~ - :
.' , 2~n ~
,, , ~ ~
Starting nucleoside I (930 my) and imidazole (1.188 g) were dissolved in 10 ml anhydrous DMF under Argon. To this was added chlorotriisopropylsilane (1.5 ml) and the mixture was stirred at room temperature.
TLC (C~Cl2/MeO~I 9:1) after 1 hour indicated very little reaction. The mixture was allowed to stir for 22 hours. TLC of the reaction mixture (CH2Clz:MeOH:
[:9.5:0.5]) showed only a trace amount of starting material left. The mixture was stirred for another 2 hours, diluted with Et20 (30 ml) and washed with saturated NaCl (4x20 ml). The Et20 layer was dried and the solvent was removed in vacuo. The residue was purified by flash chromatography using Cll2Cl2:MeOH
(9.6:0.4). Yield = 671 mg. II
Step 2: Preparation of 5'-DMT-3'-TIPSI-N2-isobutyryl-2'-deoxyguanosine-N3-cyanoborane.
A mixture of starting nucleoside II (0.455 g) and Ph3PBH2CN (1:2 ratio respectively) in anhydrous THF
(35 ml) under argon was heated at reflux for 3 hours.
The reaction mixture was cooled in ice and then the solvent was removed in vacuo. The residue was purified , by flash chromatography on silica gel using CH2Cl2:Acetone (9.5:0.5). Fractions 24-60 (=20 ml each) contained pure product and were combined. The solvent was removed under reduced pressure. Yield = 0.163 g.
III
Step 3: Preparation of 5'-DMT-N2-isobutyryl-2'-deoxyguanosine-N~-cyanoborane.
Startinq nucleoside III (0.163 g) was ,~
dissolved in THF. To this was added 1 equivalent of n8u~NF (l.lM solution in THF) and the mixture was stirred at room temperature. After 40 min. the reaction was checked by TLC (CH2Cl2:MeOH [91:9]), which indicated that only a small amount of starting material had reacted to give the product. After 2~ hours, TLC
, - , , , ~ , : :
- ~: .. ..
-' ~' ' . .' ' ~ .
~ V ~ 0 s~owed an increase in amount of product, but starting material was th more intense spot. After 4~ hours, TLC
was similar to that after 2~ hours so another 180 ~1 of nBu,NF was added and the mixture was stirred at r.t. for qO min. TLC of the reaction mixture indicated almost complete reaction. The solvent was removed from the reaction mixture under reduced pressure. The residue was purified by flash chromatography on silica using C~zCl2:MeOH (9.4:0.6). Yield = 110 mg. IV.
lo Step 4: Preparation of N2-isobutyryl-5'-DMT-N -cyanoborane-2'-deoxyguanosine-3'(methyldiisopropyl) phosphoramidite.
Starting nucleoside IV (110 mg) in a three neck r.b. flask was evacuated and then flushed with argon. Diisopropyl ethyl amine (iPr2NEt) followed ~y CH2Cl2 were added. After complete dissolution of nucleoside, diisopropylmethyl phosphonamidic chloride (47 ~1) was added dropwise. The mixture was stirred at r.t. After 25 min. a few drops of reaction mixture -~
were taken in a mixture of EtOAc/water. TLC of the -EtOAc solution was was developed in EtOAc:CH2Cl2:Et,N --(4.5:4.5:1). Only a trace amount of reaction had occurred so another 1 equiv. of phosphonoamidic chloride was added and the mixture was allowed to stir.
After 2~ hours, only a trace amount of reaction had occurred. After 4~ hours, another equivalent of amidic chloride was added. After 7 hours, = 0.1 ml of MeOH
was added and the solution was diluted with EtOAc: 7-8 ml. It was washed with = 10% Nal~CO~ (2 x 5 ml) and sat.
NaCl (2 x 5 ml). The organic layer was dried and solvent was removed under reduced pressure.
Purification was attempted by flash chromatography using CH2Cl2:MeOH (97:3). The major spots eluted with other impurities. The product was recolumned using CH2Cl2:EtOAc (8:2). It still did not come out as a single or even two spots (for two diasteromers), .
.
, : - , .. . .
: ' ` - .:
. .
~ ~ l l J ~ O
instead 3-~ more polar spots ~although in small amounts) were present. The "P nmr shows less than 5%
impurities and ~i~ nmr of this material shows no impurit~
other than solvents used for column. The product was further dried in vacuo. Yield 11.0 mg V.
The foregoing examples are illustrative of the present invention, and are not to be taken as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.
: - .. . : , . .
- : , . . .
. . : . :
.
- : . , , ~-.. :
. .
.. : . . . . '~
,. . ~ .
transformed stain of Rous sarcoma virus, J.
Lutton et al., J. Biol. Chem. 254, 11181 (lS79) (grown in DMEM-H + 10% FCS).
The cytotoxic screens were conducted ~-according to NIH protocols, see E. Huang et al., J.
Pharm. Sci. 61, 108 (1972), with 10' cells, growth medium, antibiotics and drugs from 0.5 to 100 ~g/ml final concentration. For the L1210, l~ela-S, amd Tmolt, (i.e. the suspended cells), the incubations were conducted in sterile test tubes in a final volume of 1 ml for 72 hr at 37 C in a CO2 incubator. The cells on -the third day were still in log growth phase. The number of cells/ml are determined using trypan blue exclusion and a hemocytometer. See. e.g., R. Geran, sup~. For solid tumors there is plated out in wells 1 x 10' cells with 1 ml of medium + antibiotics and the other components of growth. When the controls have converged (~95%) then the density of the cells is estimated and the ED,o values calculated. These structures are given in Table I below, and these data are given in Table II below.
. . .
TABLE I
8tructure of_Compounds Deox~ribose Nucleoside 8ase 3' 5' -I - 7 3b Ino OH OH H BH2CN
2b InoSi(CH(CH,)2)3 Si(CH(CH,)2), 1{ 8H2CN
3a Gua OH OH ~{ BH2CN
102a GuaSi(CH(CH3) 2) ~ Si (cH(CH3) 2) 3 H BH2CN
3c Ade OH OH BH2CN H
2c AdeSi(CH(CH3) 2) 3 Si(~(C}I,) 2) 3 BH2CN 1l N~
3d Cyt OH OH BH2CN --152d CytSi(CH(CH3)2)3 Si(CH(CH3)2)3 BH2CN __ 4. CytH Si(CH(CH3) 2 ) 3 BH2CN - -TABLE II
Cytotoxicity of Boron Adductq of ribonucleosides Adeno Lung Carci- Bron- Osteo noma cho- sar-C~d. L1210 P388 Tmolt SW480 KB qenic Hela-S coma Glioma 3b 4.00 7.21 1.73 3.86 3.45 4.93 2.41 7.28 8.49 2b 4.00 7.31 6.49 2.57 5.29 6.20 3.26 NA 6.21 3a 5.08 5.87 2a 6.05 5.13 2.26 2.67 2.67 4.05 2.83 7.20 7.61 3c 3.68 5.96 1.36 2.82 3.12 5.26 2.35 7.39 6.88 2c 6.86 7.46 1.88 2.87 5.79 5.58 3.42 NA 4.99 3d 2.61 3.17 1.13 2.96 5.46 3.73 3.37 4.63 4.39 --2d 4.78 7.26 3.36 2.73 5.72 6.60 2.94 NA 6.77 4. 2.92 -- 2.34 2.71 4.07 5.58 3.63 6.62 3.~3 2n7~ 9~
EXAMPLE IlI
AntihyperliDidemic Activitv Compounds to be tested were suspended in 1%
aqueous carboxymethylcellulose, homogenized and administered to male CF~ mice (25 g) intraperitoneally for 16d. On days 9 and 16, blood was obtained by tail vein bleeding, and the serum was separated by centrifugation for three minutes. The serum cholesterol levels were determined by a modification of the Liebermann-Burchard reaction in accordance with known techniques. See A. Ness, et al., Clin. Chim.
Acta 10, 229 (1964). Serum triglyceride levels were determined with a commercial kit, the Fisher llycel Triglyceride Test Kit, for a different group of animals bled on day 16. The results of these antihyperlipidemic screens, for a compound dose of 8 mg/kg, are shown in Table III below.
EXAMPLE IV
Anti-Inflammatory Activity CF, male mice (-25 g) were administered test drugs at 5-40 mg/kg in 0.05% Tween 80-H20 intraperitoneally 3 hours prior to and 30 minutes prior to the injection of 0.2 ml of 1% carraqeenan in 0.9%
saline into the plantar surface of the right hind foot.
Saline was injected into the left hind foot, which serves as a base line. After 3 hours, both feet were excised at the tibiotarsal (ankle) joint according to standard procedures. See C. Winter et al., Proc. Soc.
Expt. Biol Med. 544, 111 (1962); A. Roszkawski et al., J._Pharm. Exp. Ther. 179, 114 (1971). Control mice afforded a 78+3 mg increase in paw weiqht. Data on the percent inhibition of the inflammatory response for a dose of 8 mg/kg are reported in Table III below.
.
. . . .. ..
'' , : ' . - ~ ' , .~
. ' . ~ . , .
.~; :' ' TA~LE III
Antiinflammatory and }Iypolipidemic ~ctivities of ~oron-Containinq Nucleo~3ide~3 Antiinflammatory llvpolipidemic Cholesterol Triglyceride Activity at Inhibition at Inhibition at 8 mg/kg/day 8 mg/kg/day 8 mg/kg/day Compo~lnd ~ inhibition % inhibition % inhibitlon 3a 22 44 37 lo 3b 25 19 30 3c 22 44 37 3d 52 38 23 Standard 47 (phenyl- 13 (clofib- 2S
butazone) rate at 130 mg/
~g/day)n EXAMPLE V
A. Synthesis of oligomers ~cheme II
G' G^^
20DMT--o~o ¦ DMr~)~l~o I DI~T-O ~0 H\f~H m l ~ 201- Hr\~ H ~ nDu ~ NE' H I~ ir~3Sio H lPr]Sl IllU
G^^ G'' DI~T - C~ ~¦ C l P I O~le l N I r ~ 2 ~ ~¦ . -H\ j~/H l~\~H
HO HMcO - I ~ ~ 11 r 1 ~ lll 2C~
IcH3)2cllc~-C~N~ (Cll3) 2CIICN - l T ~
Step 1 Preparation of 5'-DMT-3'-TIPSI-N2-isobutyryl-2'-deoxyguanosine.
: . . . . . ~ - :
.' , 2~n ~
,, , ~ ~
Starting nucleoside I (930 my) and imidazole (1.188 g) were dissolved in 10 ml anhydrous DMF under Argon. To this was added chlorotriisopropylsilane (1.5 ml) and the mixture was stirred at room temperature.
TLC (C~Cl2/MeO~I 9:1) after 1 hour indicated very little reaction. The mixture was allowed to stir for 22 hours. TLC of the reaction mixture (CH2Clz:MeOH:
[:9.5:0.5]) showed only a trace amount of starting material left. The mixture was stirred for another 2 hours, diluted with Et20 (30 ml) and washed with saturated NaCl (4x20 ml). The Et20 layer was dried and the solvent was removed in vacuo. The residue was purified by flash chromatography using Cll2Cl2:MeOH
(9.6:0.4). Yield = 671 mg. II
Step 2: Preparation of 5'-DMT-3'-TIPSI-N2-isobutyryl-2'-deoxyguanosine-N3-cyanoborane.
A mixture of starting nucleoside II (0.455 g) and Ph3PBH2CN (1:2 ratio respectively) in anhydrous THF
(35 ml) under argon was heated at reflux for 3 hours.
The reaction mixture was cooled in ice and then the solvent was removed in vacuo. The residue was purified , by flash chromatography on silica gel using CH2Cl2:Acetone (9.5:0.5). Fractions 24-60 (=20 ml each) contained pure product and were combined. The solvent was removed under reduced pressure. Yield = 0.163 g.
III
Step 3: Preparation of 5'-DMT-N2-isobutyryl-2'-deoxyguanosine-N~-cyanoborane.
Startinq nucleoside III (0.163 g) was ,~
dissolved in THF. To this was added 1 equivalent of n8u~NF (l.lM solution in THF) and the mixture was stirred at room temperature. After 40 min. the reaction was checked by TLC (CH2Cl2:MeOH [91:9]), which indicated that only a small amount of starting material had reacted to give the product. After 2~ hours, TLC
, - , , , ~ , : :
- ~: .. ..
-' ~' ' . .' ' ~ .
~ V ~ 0 s~owed an increase in amount of product, but starting material was th more intense spot. After 4~ hours, TLC
was similar to that after 2~ hours so another 180 ~1 of nBu,NF was added and the mixture was stirred at r.t. for qO min. TLC of the reaction mixture indicated almost complete reaction. The solvent was removed from the reaction mixture under reduced pressure. The residue was purified by flash chromatography on silica using C~zCl2:MeOH (9.4:0.6). Yield = 110 mg. IV.
lo Step 4: Preparation of N2-isobutyryl-5'-DMT-N -cyanoborane-2'-deoxyguanosine-3'(methyldiisopropyl) phosphoramidite.
Starting nucleoside IV (110 mg) in a three neck r.b. flask was evacuated and then flushed with argon. Diisopropyl ethyl amine (iPr2NEt) followed ~y CH2Cl2 were added. After complete dissolution of nucleoside, diisopropylmethyl phosphonamidic chloride (47 ~1) was added dropwise. The mixture was stirred at r.t. After 25 min. a few drops of reaction mixture -~
were taken in a mixture of EtOAc/water. TLC of the -EtOAc solution was was developed in EtOAc:CH2Cl2:Et,N --(4.5:4.5:1). Only a trace amount of reaction had occurred so another 1 equiv. of phosphonoamidic chloride was added and the mixture was allowed to stir.
After 2~ hours, only a trace amount of reaction had occurred. After 4~ hours, another equivalent of amidic chloride was added. After 7 hours, = 0.1 ml of MeOH
was added and the solution was diluted with EtOAc: 7-8 ml. It was washed with = 10% Nal~CO~ (2 x 5 ml) and sat.
NaCl (2 x 5 ml). The organic layer was dried and solvent was removed under reduced pressure.
Purification was attempted by flash chromatography using CH2Cl2:MeOH (97:3). The major spots eluted with other impurities. The product was recolumned using CH2Cl2:EtOAc (8:2). It still did not come out as a single or even two spots (for two diasteromers), .
.
, : - , .. . .
: ' ` - .:
. .
~ ~ l l J ~ O
instead 3-~ more polar spots ~although in small amounts) were present. The "P nmr shows less than 5%
impurities and ~i~ nmr of this material shows no impurit~
other than solvents used for column. The product was further dried in vacuo. Yield 11.0 mg V.
The foregoing examples are illustrative of the present invention, and are not to be taken as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.
: - .. . : , . .
- : , . . .
. . : . :
.
- : . , , ~-.. :
. .
.. : . . . . '~
,. . ~ .
Claims
That Which is Claimed is:
1. A boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
2. A boronated nucleotide comprising a 5' phosphate ester of a nucleoside according to Claim 1.
3. A boronated oligonucleotide comprising a chain of natural or modified ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a nucleotide according to Claim 2.
4. A boronated nucleoside according to Claim 1 wherein the boron-containing substituent is -BH2CN.
5. A boronated nucleoside according to Claim 1 wherein the base of said nucleoside is selected from the group consisting of adenine, cytosine, guanine, inosine, and analogs thereof.
6. A boronated nucleoside according to Claim 1 wherein all hydroxyl substituents on the ribose moiety are O-protected.
7. A boronated nucleoside according to Claim 1 wherein all hydroxyl substituents on the ribose moiety are O-protected as silyl ethers.
8. A boronated nucleoside according to Claim 1 wherein the 3' and 5' hydroxy substituents are O-protected as silyl ethers.
9. A boronated nucleoside according to Claim 1 wherein the 3' and 5' hydroxy substituents are O-protected as triisopropylsilyl ethers.
10. A boronated oligonucleotide according to Claim 3 comprising from 2 to 50 nucleotides.
11. A boronated oligonucleotide according to Claim 3 comprising from 2 to 30 nucleotides.
12. A boronated oligonucleotide according to Claim 3 comprising from 2 to 18 nucleotides.
13. A boronated nucleoside selected from the group consisting of 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2'-deoxyadenosine-N1-cyanoborane, 2'-deoxycytidine-N3-cyanoborane, and 2',3'-dideoxycytidine-N3-cyanoborane.
14. A method for synthesizing N-boronated nucleosides from a substrate nucleoside comprised of a ribose moiety covalently bound to a nitrogenous base, the method comprising:
(a) protecting the hydroxy substituents of the ribose; then (b) boronating the nitrogenous base; and then (c) deprotecting the hydroxy substituents.
19. A method for treating a tumor bearing mammal comprising administering to said mammal a therapeutically effective amount of a boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
20. A method according to Claim 19, further comprising the step of concurrently administering a tumor-combatting amount of one or more other antineoplastic agent.
21. A method for treating a tumor bearing mammal comprising:
administering to said mammal a therapeutically effective amount of a boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl; and exposing said mammal to thermal neutron radiation.
22. A method for treating a mammal suffering from inflammation comprising administering to said mammal a therapeutically effective amount of a boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
23. A method according to Claim 22, further comprising the step of concurrently administering to said mammal an inflammation-combatting amount of one or more other antiinflammatory agent.
24. A method for treating a hyperlipidemic mammal comprising administering to said mammal a therapeutically effective amount of a boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
25. A method according to Claim 24, further comprising the step of concurrently administering to said mammal a hyperlipidemia-combatting amount of one or more other antihperlipidemic agent.
26. A method according to Claim 19, 22, or 24 wherein the boronated nucleoside is selected from the group consisting of 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2'-deoxyadenosine-N1-cyanoborane, and 2'-deoxycytidine-N3-cyanoborane.
27. A pharmaceutical formulation comprising a therapeutically effective amount of a boronated nucleoside in a pharmaceutically acceptable carrier, said boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
28. A pharmaceutical formulation according to Claim 27 wherein the boronated nucleoside is selected from one or more of the group consisting of 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2-deoxyadenosine-N1-cyanoborane, and 2'-deoxycytidine-N3-cyanoborane.
29. A method according to Claim 22, wherein the boronated nucleoside is selected from the group consisting of 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2'-deoxyadenosine-N1-cyanoborane, and 2'-deoxycytidine-N3-cyanoborane.
30. A boronated nucleoside according to claim 1 selected from the group consisting of:
guanosine-N7-cyanoborane:
inosine-N7-cyanoborane;
adenosine-N1-cyanoborane;
cytidine-N3-cyanoborane;
guanosine-N7-borane;
inosine-N7-borane;
adenosine-N1-borane;
cytidine-N3-borane;
guanosine-N7-carbomethoxyborane;
inosine-N7-carboethoxyborane;
adenosine-N1-carbopropoxyborane;
cytidine-N3-carbobutoxyborane;
2'-deoxyguanosine-N7-cyanoborane;
2'-deoxyinosine-N7-cyanoborane;
2'-deoxyadenosine-N1-cyanoborane;
2'-deoxycytidine-N3-cyanoborane;
2'-deoxyguanosine-N7-borane;
2'-deoxyinosine-N7-borane;
2'-deoxyadenosine-N1-borane;
2'-deoxycytidine-N3-borane;
2'-deoxyguanosine-N7-carbomethoxyborane;
2'-deoxyinosine-N7-carboethoxyborane;
2'-deoxyadenosine-N1-carbopropoxyborane;
2'-deoxycytidine-N3-carbobutoxyborane;
2',3'-dideoxyguanosine-N7-cyanoborane;
2',3'-dideoxyinosine-N7-cyanoborane;
2',3'-dideoxyadenosine-N1-cyanoborane;
2',3'-dideoxycytidine-N3-cyanoborane;
2',3'-dideoxyguanosine-N7-borane;
2',3'-dideoxyinosine-N7-borane;
2',3'-dideoxyadenosine-N1-borane;
2',3'-dideoxycytidine-N3-borane;
2',3'-dideoxyguanosine-N7-carbomethoxyborane;
2',3'-dideoxyinosine-N7-carboethoxyborane:
2',3'-dideoxyadenosine-N1-carbopropoxyborane; and 2',3'-dideoxycytidine-N3-carbobutoxyborane.
31. A boronated oligonucleotide comprising a chain of 2 to 50 ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a nucleotide according to Claim 2.
32. A boronated oligonucleotide comprising a chain of 2 to 30 ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a nucleotide according to Claim 2.
33. A boronated oligonucleotide comprising a chain of 2 to 18 ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a nucleotide according to Claim 2.
1. A boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
2. A boronated nucleotide comprising a 5' phosphate ester of a nucleoside according to Claim 1.
3. A boronated oligonucleotide comprising a chain of natural or modified ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a nucleotide according to Claim 2.
4. A boronated nucleoside according to Claim 1 wherein the boron-containing substituent is -BH2CN.
5. A boronated nucleoside according to Claim 1 wherein the base of said nucleoside is selected from the group consisting of adenine, cytosine, guanine, inosine, and analogs thereof.
6. A boronated nucleoside according to Claim 1 wherein all hydroxyl substituents on the ribose moiety are O-protected.
7. A boronated nucleoside according to Claim 1 wherein all hydroxyl substituents on the ribose moiety are O-protected as silyl ethers.
8. A boronated nucleoside according to Claim 1 wherein the 3' and 5' hydroxy substituents are O-protected as silyl ethers.
9. A boronated nucleoside according to Claim 1 wherein the 3' and 5' hydroxy substituents are O-protected as triisopropylsilyl ethers.
10. A boronated oligonucleotide according to Claim 3 comprising from 2 to 50 nucleotides.
11. A boronated oligonucleotide according to Claim 3 comprising from 2 to 30 nucleotides.
12. A boronated oligonucleotide according to Claim 3 comprising from 2 to 18 nucleotides.
13. A boronated nucleoside selected from the group consisting of 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2'-deoxyadenosine-N1-cyanoborane, 2'-deoxycytidine-N3-cyanoborane, and 2',3'-dideoxycytidine-N3-cyanoborane.
14. A method for synthesizing N-boronated nucleosides from a substrate nucleoside comprised of a ribose moiety covalently bound to a nitrogenous base, the method comprising:
(a) protecting the hydroxy substituents of the ribose; then (b) boronating the nitrogenous base; and then (c) deprotecting the hydroxy substituents.
19. A method for treating a tumor bearing mammal comprising administering to said mammal a therapeutically effective amount of a boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
20. A method according to Claim 19, further comprising the step of concurrently administering a tumor-combatting amount of one or more other antineoplastic agent.
21. A method for treating a tumor bearing mammal comprising:
administering to said mammal a therapeutically effective amount of a boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl; and exposing said mammal to thermal neutron radiation.
22. A method for treating a mammal suffering from inflammation comprising administering to said mammal a therapeutically effective amount of a boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
23. A method according to Claim 22, further comprising the step of concurrently administering to said mammal an inflammation-combatting amount of one or more other antiinflammatory agent.
24. A method for treating a hyperlipidemic mammal comprising administering to said mammal a therapeutically effective amount of a boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
25. A method according to Claim 24, further comprising the step of concurrently administering to said mammal a hyperlipidemia-combatting amount of one or more other antihperlipidemic agent.
26. A method according to Claim 19, 22, or 24 wherein the boronated nucleoside is selected from the group consisting of 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2'-deoxyadenosine-N1-cyanoborane, and 2'-deoxycytidine-N3-cyanoborane.
27. A pharmaceutical formulation comprising a therapeutically effective amount of a boronated nucleoside in a pharmaceutically acceptable carrier, said boronated nucleoside comprising a nucleoside which is N-boronated on the nucleoside base with a boron-containing substituent selected from the group consisting of -BH2CN, -BH3, and -BH2COOR, wherein R is C1 to C18 alkyl.
28. A pharmaceutical formulation according to Claim 27 wherein the boronated nucleoside is selected from one or more of the group consisting of 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2-deoxyadenosine-N1-cyanoborane, and 2'-deoxycytidine-N3-cyanoborane.
29. A method according to Claim 22, wherein the boronated nucleoside is selected from the group consisting of 2'-deoxyguanosine-N7-cyanoborane, 2'-deoxyinosine-N7-cyanoborane, 2'-deoxyadenosine-N1-cyanoborane, and 2'-deoxycytidine-N3-cyanoborane.
30. A boronated nucleoside according to claim 1 selected from the group consisting of:
guanosine-N7-cyanoborane:
inosine-N7-cyanoborane;
adenosine-N1-cyanoborane;
cytidine-N3-cyanoborane;
guanosine-N7-borane;
inosine-N7-borane;
adenosine-N1-borane;
cytidine-N3-borane;
guanosine-N7-carbomethoxyborane;
inosine-N7-carboethoxyborane;
adenosine-N1-carbopropoxyborane;
cytidine-N3-carbobutoxyborane;
2'-deoxyguanosine-N7-cyanoborane;
2'-deoxyinosine-N7-cyanoborane;
2'-deoxyadenosine-N1-cyanoborane;
2'-deoxycytidine-N3-cyanoborane;
2'-deoxyguanosine-N7-borane;
2'-deoxyinosine-N7-borane;
2'-deoxyadenosine-N1-borane;
2'-deoxycytidine-N3-borane;
2'-deoxyguanosine-N7-carbomethoxyborane;
2'-deoxyinosine-N7-carboethoxyborane;
2'-deoxyadenosine-N1-carbopropoxyborane;
2'-deoxycytidine-N3-carbobutoxyborane;
2',3'-dideoxyguanosine-N7-cyanoborane;
2',3'-dideoxyinosine-N7-cyanoborane;
2',3'-dideoxyadenosine-N1-cyanoborane;
2',3'-dideoxycytidine-N3-cyanoborane;
2',3'-dideoxyguanosine-N7-borane;
2',3'-dideoxyinosine-N7-borane;
2',3'-dideoxyadenosine-N1-borane;
2',3'-dideoxycytidine-N3-borane;
2',3'-dideoxyguanosine-N7-carbomethoxyborane;
2',3'-dideoxyinosine-N7-carboethoxyborane:
2',3'-dideoxyadenosine-N1-carbopropoxyborane; and 2',3'-dideoxycytidine-N3-carbobutoxyborane.
31. A boronated oligonucleotide comprising a chain of 2 to 50 ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a nucleotide according to Claim 2.
32. A boronated oligonucleotide comprising a chain of 2 to 30 ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a nucleotide according to Claim 2.
33. A boronated oligonucleotide comprising a chain of 2 to 18 ribonucleotides or deoxyribonucleotides, at least one nucleotide of said oligonucleotide comprising a nucleotide according to Claim 2.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US453,311 | 1989-12-20 | ||
US07/453,311 US5130302A (en) | 1989-12-20 | 1989-12-20 | Boronated nucleoside, nucleotide and oligonucleotide compounds, compositions and methods for using same |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2071936A1 true CA2071936A1 (en) | 1991-06-21 |
Family
ID=23800042
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002071936A Abandoned CA2071936A1 (en) | 1989-12-20 | 1990-12-17 | Boronated nucleosides |
Country Status (8)
Country | Link |
---|---|
US (1) | US5130302A (en) |
EP (1) | EP0506892B1 (en) |
JP (1) | JPH05507265A (en) |
AT (1) | ATE159027T1 (en) |
AU (1) | AU634450B2 (en) |
CA (1) | CA2071936A1 (en) |
DE (1) | DE69031567D1 (en) |
WO (1) | WO1991009048A1 (en) |
Families Citing this family (644)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5177198A (en) * | 1989-11-30 | 1993-01-05 | University Of N.C. At Chapel Hill | Process for preparing oligoribonucleoside and oligodeoxyribonucleoside boranophosphates |
US5362732A (en) * | 1989-12-20 | 1994-11-08 | University Of North Carolina At Chapel Hill | Boronated compounds |
US5260427A (en) * | 1991-05-10 | 1993-11-09 | Boron Biolgicals, Inc. | Nucleosidylphosphite-borane compounds and method of making the same |
US5545397A (en) * | 1991-10-23 | 1996-08-13 | Boron Biologicals, Inc. | Contrast agents and compositions for radiological imaging, and radiological imaging method utilizing same |
US5256394A (en) * | 1991-10-23 | 1993-10-26 | Boron Biologicals, Inc. | Radiological imaging method, and contrast media reagents therefor |
US6335434B1 (en) | 1998-06-16 | 2002-01-01 | Isis Pharmaceuticals, Inc., | Nucleosidic and non-nucleosidic folate conjugates |
US8153602B1 (en) | 1991-11-19 | 2012-04-10 | Isis Pharmaceuticals, Inc. | Composition and methods for the pulmonary delivery of nucleic acids |
US5405598A (en) * | 1992-02-24 | 1995-04-11 | Schinazi; Raymond F. | Sensitizing agents for use in boron neutron capture therapy |
US5466679A (en) * | 1993-05-17 | 1995-11-14 | The Ohio State University Research Foundation | Carboranyl uridines and their use in boron neutron capture therapy |
US5859231A (en) * | 1993-09-03 | 1999-01-12 | Duke University | Synthesis of oligonucleotides with boranophosphonate linkages |
WO1995006752A1 (en) * | 1993-09-03 | 1995-03-09 | Duke University | A method of nucleic acid sequencing |
DE69433036T2 (en) | 1993-09-03 | 2004-05-27 | Isis Pharmaceuticals, Inc., Carlsbad | AMINODERIVATIZED NUCLEOSIDES AND OLIGONUCLEOSIDES |
US6376178B1 (en) | 1993-09-03 | 2002-04-23 | Duke University | Method of nucleic acid sequencing |
US6180766B1 (en) * | 1993-12-02 | 2001-01-30 | Raymond F. Schinazi | Nucleosides and oligonucleotides containing boron clusters |
US5599796A (en) * | 1993-12-02 | 1997-02-04 | Emory University | Treatment of urogenital cancer with boron neutron capture therapy |
US5643893A (en) * | 1994-06-22 | 1997-07-01 | Macronex, Inc. | N-substituted-(Dihydroxyboryl)alkyl purine, indole and pyrimidine derivatives, useful as inhibitors of inflammatory cytokines |
US5550132A (en) * | 1994-06-22 | 1996-08-27 | University Of North Carolina | Hydroxyalkylammonium-pyrimidines or purines and nucleoside derivatives, useful as inhibitors of inflammatory cytokines |
US5595878A (en) * | 1995-06-02 | 1997-01-21 | Boron Biologicals, Inc. | Detection of biopolymers and biooligomers with boron hydride labels |
US6420549B1 (en) | 1995-06-06 | 2002-07-16 | Isis Pharmaceuticals, Inc. | Oligonucleotide analogs having modified dimers |
US5854033A (en) | 1995-11-21 | 1998-12-29 | Yale University | Rolling circle replication reporter systems |
EP0932698A1 (en) * | 1996-03-26 | 1999-08-04 | Lynx Therapeutics, Inc. | Oligonucleotide treatments and compositions for human melanoma |
US5898031A (en) | 1996-06-06 | 1999-04-27 | Isis Pharmaceuticals, Inc. | Oligoribonucleotides for cleaving RNA |
US20030044941A1 (en) | 1996-06-06 | 2003-03-06 | Crooke Stanley T. | Human RNase III and compositions and uses thereof |
US9096636B2 (en) * | 1996-06-06 | 2015-08-04 | Isis Pharmaceuticals, Inc. | Chimeric oligomeric compounds and their use in gene modulation |
US20040161777A1 (en) * | 1996-06-06 | 2004-08-19 | Baker Brenda F. | Modified oligonucleotides for use in RNA interference |
US20070275921A1 (en) * | 1996-06-06 | 2007-11-29 | Isis Pharmaceuticals, Inc. | Oligomeric Compounds That Facilitate Risc Loading |
US20040266706A1 (en) * | 2002-11-05 | 2004-12-30 | Muthiah Manoharan | Cross-linked oligomeric compounds and their use in gene modulation |
US7812149B2 (en) | 1996-06-06 | 2010-10-12 | Isis Pharmaceuticals, Inc. | 2′-Fluoro substituted oligomeric compounds and compositions for use in gene modulations |
US6111085A (en) * | 1996-09-13 | 2000-08-29 | Isis Pharmaceuticals, Inc. | Carbamate-derivatized nucleosides and oligonucleosides |
US6444659B1 (en) * | 1996-11-28 | 2002-09-03 | Cognis Deutschland Gmbh | Use of mixtures of active substances, containing phytostenols and/or phytostenol esters and potentiators, for the production of hypocholesterolemic agents |
AU731909B2 (en) | 1997-07-01 | 2001-04-05 | Isis Pharmaceuticals, Inc. | Compositions and methods for the delivery of oligonucleotides via the alimentary canal |
US7321828B2 (en) | 1998-04-13 | 2008-01-22 | Isis Pharmaceuticals, Inc. | System of components for preparing oligonucleotides |
AU745880B2 (en) | 1998-05-21 | 2002-04-11 | Isis Pharmaceuticals, Inc. | Compositions and methods for non-parenteral delivery of oligonucleotides |
JP2002515514A (en) | 1998-05-21 | 2002-05-28 | アイシス・ファーマシューティカルス・インコーポレーテッド | Compositions and methods for local delivery of oligonucleotides |
US6867294B1 (en) | 1998-07-14 | 2005-03-15 | Isis Pharmaceuticals, Inc. | Gapped oligomers having site specific chiral phosphorothioate internucleoside linkages |
US6225293B1 (en) | 1998-09-02 | 2001-05-01 | Isis Pharmaceuticals, Inc. | Methods and compounds for tracking the biodistribution of macromolecule-carrier combinations |
US6077709A (en) | 1998-09-29 | 2000-06-20 | Isis Pharmaceuticals Inc. | Antisense modulation of Survivin expression |
US6300320B1 (en) | 1999-01-05 | 2001-10-09 | Isis Pharmaceuticals, Inc. | Modulation of c-jun using inhibitors of protein kinase C |
US6127124A (en) * | 1999-01-20 | 2000-10-03 | Isis Pharmaceuticals, Inc. | Fluorescence based nuclease assay |
US7098192B2 (en) | 1999-04-08 | 2006-08-29 | Isis Pharmaceuticals, Inc. | Antisense oligonucleotide modulation of STAT3 expression |
CN1370170A (en) * | 1999-05-25 | 2002-09-18 | 宾夕法尼亚州研究基金会 | DNA methyltransferase inhibitors |
EP1420021A1 (en) * | 1999-05-25 | 2004-05-19 | The Penn State Research Foundation | DNA Methyltransferase inhibitors |
US6656730B1 (en) | 1999-06-15 | 2003-12-02 | Isis Pharmaceuticals, Inc. | Oligonucleotides conjugated to protein-binding drugs |
US6147200A (en) * | 1999-08-19 | 2000-11-14 | Isis Pharmaceuticals, Inc. | 2'-O-acetamido modified monomers and oligomers |
US6261840B1 (en) | 2000-01-18 | 2001-07-17 | Isis Pharmaceuticals, Inc. | Antisense modulation of PTP1B expression |
US20020055479A1 (en) | 2000-01-18 | 2002-05-09 | Cowsert Lex M. | Antisense modulation of PTP1B expression |
JP2003530841A (en) * | 2000-04-13 | 2003-10-21 | トーマス エヌ. ワイト, | Therapeutic compounds and methods |
US6680172B1 (en) | 2000-05-16 | 2004-01-20 | Regents Of The University Of Michigan | Treatments and markers for cancers of the central nervous system |
US20060166227A1 (en) * | 2000-06-20 | 2006-07-27 | Stephen Kingsmore | Protein expression profiling |
US8568766B2 (en) | 2000-08-24 | 2013-10-29 | Gattadahalli M. Anantharamaiah | Peptides and peptide mimetics to treat pathologies associated with eye disease |
WO2002030465A2 (en) | 2000-10-12 | 2002-04-18 | University Of Rochester | Compositions that inhibit proliferation of cancer cells |
US7767802B2 (en) | 2001-01-09 | 2010-08-03 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of anti-apoptotic genes |
US6573051B2 (en) * | 2001-03-09 | 2003-06-03 | Molecular Staging, Inc. | Open circle probes with intramolecular stem structures |
ATE434936T1 (en) | 2001-03-14 | 2009-07-15 | Myriad Genetics Inc | TSG101-GAG INTERACTION AND THEIR USE |
US20070015144A9 (en) * | 2001-05-25 | 2007-01-18 | Genset, S.A. | Human cDNAs and proteins and uses thereof |
DK2000545T3 (en) | 2001-06-20 | 2011-11-28 | Genentech Inc | Compositions and methods for diagnosis and treatment of lung tumor |
US7803915B2 (en) | 2001-06-20 | 2010-09-28 | Genentech, Inc. | Antibody compositions for the diagnosis and treatment of tumor |
CA2451643C (en) | 2001-06-21 | 2012-11-13 | Isis Pharmaceuticals, Inc. | Antisense modulation of superoxide dismutase 1, soluble expression |
US7425545B2 (en) | 2001-07-25 | 2008-09-16 | Isis Pharmaceuticals, Inc. | Modulation of C-reactive protein expression |
US6964950B2 (en) | 2001-07-25 | 2005-11-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of C-reactive protein expression |
US20030096772A1 (en) | 2001-07-30 | 2003-05-22 | Crooke Rosanne M. | Antisense modulation of acyl CoA cholesterol acyltransferase-2 expression |
US7407943B2 (en) | 2001-08-01 | 2008-08-05 | Isis Pharmaceuticals, Inc. | Antisense modulation of apolipoprotein B expression |
US7227014B2 (en) | 2001-08-07 | 2007-06-05 | Isis Pharmaceuticals, Inc. | Antisense modulation of apolipoprotein (a) expression |
EP2174953A1 (en) | 2001-09-18 | 2010-04-14 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
EP1488233A4 (en) | 2001-10-09 | 2006-06-21 | Genentech Inc | Novel acidic mammalian proteins and polynucleotides encoding the same |
US6750019B2 (en) | 2001-10-09 | 2004-06-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of insulin-like growth factor binding protein 5 expression |
NZ566396A (en) | 2001-10-09 | 2009-07-31 | Isis Pharmaceuticals Inc | Antisense modulation of insulin-like growth factor binding protein 5 expressions |
US20050227933A1 (en) * | 2001-11-29 | 2005-10-13 | Benkovic Stephen J | Treatment of bacterial induced diseases using DNA methyl transferase inhibitors |
US6965025B2 (en) | 2001-12-10 | 2005-11-15 | Isis Pharmaceuticals, Inc. | Antisense modulation of connective tissue growth factor expression |
US7255874B1 (en) | 2001-12-21 | 2007-08-14 | Closure Medical Corporation | Biocompatible polymers and adhesives: compositions, methods of making and uses related thereto |
AU2002367318B2 (en) | 2002-01-02 | 2007-07-12 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
JP4138662B2 (en) * | 2002-01-10 | 2008-08-27 | ザ・ペンシルバニア・ステイト・リサーチ・フアウンデイシヨン | Process for preparing alkyl diaryl borinates and complexed diaryl boronic acids. |
US7553619B2 (en) | 2002-02-08 | 2009-06-30 | Qiagen Gmbh | Detection method using dissociated rolling circle amplification |
EP1485395A4 (en) * | 2002-02-28 | 2011-04-13 | Biota Scient Management | Nucleotide mimics and their prodrugs |
US20030180712A1 (en) | 2002-03-20 | 2003-09-25 | Biostratum Ab | Inhibition of the beta3 subunit of L-type Ca2+ channels |
US7169916B2 (en) * | 2002-04-01 | 2007-01-30 | Isis Pharmaceuticals, Inc. | Chloral-free DCA in oligonucleotide synthesis |
JP2005536190A (en) | 2002-04-16 | 2005-12-02 | ジェネンテック・インコーポレーテッド | Compositions and methods for tumor diagnosis and treatment |
WO2003097643A1 (en) * | 2002-05-17 | 2003-11-27 | Neurogen Corporation | Substituted ring-fused imidazole derivates: gabaa receptor ligands |
US7199107B2 (en) | 2002-05-23 | 2007-04-03 | Isis Pharmaceuticals, Inc. | Antisense modulation of kinesin-like 1 expression |
US20040092470A1 (en) * | 2002-06-18 | 2004-05-13 | Leonard Sherry A. | Dry powder oligonucleotide formualtion, preparation and its uses |
EP2116604A1 (en) | 2002-08-05 | 2009-11-11 | University of Rochester | Protein transducing domain/deaminase chimeric proteins, related compounds, and uses thereof |
US20050196382A1 (en) * | 2002-09-13 | 2005-09-08 | Replicor, Inc. | Antiviral oligonucleotides targeting viral families |
CN1694959B (en) * | 2002-09-13 | 2013-09-18 | 雷普利瑟公司 | Non-sequence complementary antiviral oligonucleotides |
EP2322535A3 (en) | 2002-09-20 | 2011-09-28 | Yale University | Riboswitches, methods for their use, and compositions for use with riboswitches |
US7229976B2 (en) | 2002-09-26 | 2007-06-12 | Isis Pharmaceuticals, Inc. | Modulation of forkhead box O1A expression |
US9150606B2 (en) * | 2002-11-05 | 2015-10-06 | Isis Pharmaceuticals, Inc. | Compositions comprising alternating 2'-modified nucleosides for use in gene modulation |
AU2003290598A1 (en) | 2002-11-05 | 2004-06-03 | Isis Pharmaceuticals, Inc. | Modified oligonucleotides for use in rna interference |
US9150605B2 (en) | 2002-11-05 | 2015-10-06 | Isis Pharmaceuticals, Inc. | Compositions comprising alternating 2′-modified nucleosides for use in gene modulation |
AU2003287505A1 (en) | 2002-11-05 | 2004-06-03 | Isis Pharmaceuticals, Inc. | Chimeric oligomeric compounds and their use in gene modulation |
US7696345B2 (en) | 2002-11-05 | 2010-04-13 | Isis Pharmaceuticals, Inc. | Polycyclic sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation |
ES2420914T3 (en) | 2002-11-13 | 2013-08-27 | Genzyme Corporation | Antisense modulation of apolipoprotein B expression |
CA2505801A1 (en) | 2002-11-13 | 2004-05-27 | Rosanne Crooke | Antisense modulation of apolipoprotein b expression |
CA2506127C (en) | 2002-11-15 | 2013-07-09 | Morphotek, Inc. | Methods of generating high-production of antibodies from hybridomas created by in vitro immunization |
US8007804B2 (en) | 2002-11-15 | 2011-08-30 | Musc Foundation For Research Development | Complement receptor 2 targeted complement modulators |
EP2410332A1 (en) | 2002-11-21 | 2012-01-25 | The University Of Utah | Method for identifying purinergic modulators of the olfactory system |
US7144999B2 (en) | 2002-11-23 | 2006-12-05 | Isis Pharmaceuticals, Inc. | Modulation of hypoxia-inducible factor 1 alpha expression |
WO2004058987A2 (en) | 2002-12-20 | 2004-07-15 | Qiagen Gmbh | Nucleic acid amplification |
US9487823B2 (en) | 2002-12-20 | 2016-11-08 | Qiagen Gmbh | Nucleic acid amplification |
US6977153B2 (en) * | 2002-12-31 | 2005-12-20 | Qiagen Gmbh | Rolling circle amplification of RNA |
US7468356B2 (en) | 2003-02-11 | 2008-12-23 | Antisense Therapeutics Ltd. | Modulation of insulin like growth factor I receptor expression |
US7002006B2 (en) * | 2003-02-12 | 2006-02-21 | Isis Pharmaceuticals, Inc. | Protection of nucleosides |
US7803781B2 (en) | 2003-02-28 | 2010-09-28 | Isis Pharmaceuticals, Inc. | Modulation of growth hormone receptor expression and insulin-like growth factor expression |
ATE479752T1 (en) | 2003-03-07 | 2010-09-15 | Alnylam Pharmaceuticals Inc | THERAPEUTIC COMPOSITIONS |
US20040185559A1 (en) | 2003-03-21 | 2004-09-23 | Isis Pharmaceuticals Inc. | Modulation of diacylglycerol acyltransferase 1 expression |
US8043834B2 (en) | 2003-03-31 | 2011-10-25 | Qiagen Gmbh | Universal reagents for rolling circle amplification and methods of use |
WO2004091515A2 (en) | 2003-04-09 | 2004-10-28 | Alnylam Pharmaceuticals, Inc. | iRNA CONJUGATES |
US7598227B2 (en) | 2003-04-16 | 2009-10-06 | Isis Pharmaceuticals Inc. | Modulation of apolipoprotein C-III expression |
EP1625138A4 (en) | 2003-04-17 | 2010-06-23 | Alnylam Pharmaceuticals Inc | Protected monomers |
EP2660322A3 (en) | 2003-04-17 | 2013-11-13 | Alnylam Pharmaceuticals Inc. | Modified iRNA agents |
US7399853B2 (en) | 2003-04-28 | 2008-07-15 | Isis Pharmaceuticals | Modulation of glucagon receptor expression |
US7276599B2 (en) * | 2003-06-02 | 2007-10-02 | Isis Pharmaceuticals, Inc. | Oligonucleotide synthesis with alternative solvents |
WO2005002507A2 (en) | 2003-06-03 | 2005-01-13 | Isis Pharmaceuticals, Inc. | Modulation of survivin expression |
US7786290B2 (en) | 2003-06-13 | 2010-08-31 | Alnylam Pharmaceuticals, Inc. | Double-stranded ribonucleic acid with increased effectiveness in an organism |
CA2533701A1 (en) | 2003-07-31 | 2005-02-17 | Isis Pharmaceuticals, Inc. | Oligomeric compounds and compositions for use in modulation of small non-coding rnas |
US7825235B2 (en) | 2003-08-18 | 2010-11-02 | Isis Pharmaceuticals, Inc. | Modulation of diacylglycerol acyltransferase 2 expression |
PT1660057E (en) | 2003-08-27 | 2012-08-02 | Ophthotech Corp | Combination therapy for the treatment of ocular neovascular disorders |
US20050053981A1 (en) * | 2003-09-09 | 2005-03-10 | Swayze Eric E. | Gapped oligomeric compounds having linked bicyclic sugar moieties at the termini |
US20070123480A1 (en) * | 2003-09-11 | 2007-05-31 | Replicor Inc. | Oligonucleotides targeting prion diseases |
NZ545134A (en) | 2003-09-18 | 2009-06-26 | Lilly Co Eli | Modulation of eIF4E expression |
AU2004274021B2 (en) * | 2003-09-18 | 2009-08-13 | Isis Pharmaceuticals, Inc. | 4'-thionucleosides and oligomeric compounds |
EP1678194B1 (en) | 2003-10-10 | 2013-06-26 | Alchemia Oncology Pty Limited | The modulation of hyaluronan synthesis and degradation in the treatment of disease |
US20050191653A1 (en) | 2003-11-03 | 2005-09-01 | Freier Susan M. | Modulation of SGLT2 expression |
ES2493016T3 (en) | 2003-11-17 | 2014-09-11 | Genentech, Inc. | Compositions comprising antibodies against CD79b conjugated to a growth inhibitory agent or a cytotoxic agent and methods for the treatment of tumors of hematopoietic origin |
JP2007520222A (en) | 2004-01-20 | 2007-07-26 | アイシス ファーマシューティカルズ インコーポレイテッド | Regulation of glucocorticoid receptor expression |
US7468431B2 (en) | 2004-01-22 | 2008-12-23 | Isis Pharmaceuticals, Inc. | Modulation of eIF4E-BP2 expression |
US8569474B2 (en) | 2004-03-09 | 2013-10-29 | Isis Pharmaceuticals, Inc. | Double stranded constructs comprising one or more short strands hybridized to a longer strand |
US8790919B2 (en) | 2004-03-15 | 2014-07-29 | Isis Pharmaceuticals, Inc. | Compositions and methods for optimizing cleavage of RNA by RNase H |
US20050244869A1 (en) | 2004-04-05 | 2005-11-03 | Brown-Driver Vickie L | Modulation of transthyretin expression |
EP2540734B1 (en) | 2004-04-05 | 2016-03-30 | Alnylam Pharmaceuticals, Inc. | Process and reagents for oligonucleotide synthesis and purification |
US20050260755A1 (en) * | 2004-04-06 | 2005-11-24 | Isis Pharmaceuticals, Inc. | Sequential delivery of oligomeric compounds |
EP1750776A2 (en) | 2004-04-30 | 2007-02-14 | Alnylam Pharmaceuticals Inc. | Oligonucleotides comprising a c5-modified pyrimidine |
DK1773872T3 (en) | 2004-05-21 | 2017-05-08 | Uab Res Found | VARIABLE Lymphocyte Receptors, Associated Polypeptides and Nucleic Acids, and Uses thereof |
US8394947B2 (en) | 2004-06-03 | 2013-03-12 | Isis Pharmaceuticals, Inc. | Positionally modified siRNA constructs |
US20090048192A1 (en) * | 2004-06-03 | 2009-02-19 | Isis Pharmaceuticals, Inc. | Double Strand Compositions Comprising Differentially Modified Strands for Use in Gene Modulation |
JP5192234B2 (en) | 2004-08-10 | 2013-05-08 | アルナイラム ファーマシューティカルズ, インコーポレイテッド | Chemically modified oligonucleotide |
US7427675B2 (en) | 2004-08-23 | 2008-09-23 | Isis Pharmaceuticals, Inc. | Compounds and methods for the characterization of oligonucleotides |
US7884086B2 (en) | 2004-09-08 | 2011-02-08 | Isis Pharmaceuticals, Inc. | Conjugates for use in hepatocyte free uptake assays |
EP1799859B1 (en) | 2004-09-17 | 2014-07-02 | Isis Pharmaceuticals, Inc. | Enhanced antisense oligonucleotides |
WO2006032144A1 (en) * | 2004-09-23 | 2006-03-30 | Arc Pharmaceuticals, Inc. | Pharmaceutical compositions and methods relating to inhibiting fibrous adhesions or inflammatory disease using fucans from various echinoderm sources |
AU2006223498A1 (en) | 2005-03-10 | 2006-09-21 | Genentech, Inc. | Methods and compositions for modulating vascular integrity |
US7476733B2 (en) | 2005-03-25 | 2009-01-13 | The United States Of America As Represented By The Department Of Health And Human Services | Development of a real-time PCR assay for detection of pneumococcal DNA and diagnosis of pneumococccal disease |
EP1863908B1 (en) | 2005-04-01 | 2010-11-17 | Qiagen GmbH | Reverse transcription and amplification of rna with simultaneous degradation of dna |
JP5329949B2 (en) | 2005-05-31 | 2013-10-30 | エコーレ ポリテクニーク フェデラーレ デ ローザンヌ | Triblock copolymers for cytoplasmic delivery of gene-based drugs |
US8252756B2 (en) | 2005-06-14 | 2012-08-28 | Northwestern University | Nucleic acid functionalized nanoparticles for therapeutic applications |
US20090270479A1 (en) * | 2005-07-12 | 2009-10-29 | Antonio Giordano | Genetic and Epigenetic Alterations In the Diagnosis and Treatment of Cancer |
EP1929012B1 (en) | 2005-08-11 | 2010-10-06 | Synthetic Genomics, Inc. | Method for in vitro recombination |
AU2006281569A1 (en) | 2005-08-17 | 2007-02-22 | Medexis S.A. | Composition and method for determination of CK19 expression |
WO2007027775A2 (en) | 2005-08-29 | 2007-03-08 | Isis Pharmaceuticals, Inc. | Methods for use in modulating mir-122a |
EP1762627A1 (en) | 2005-09-09 | 2007-03-14 | Qiagen GmbH | Method for the activation of a nucleic acid for performing a polymerase reaction |
EA200800868A1 (en) | 2005-09-19 | 2008-10-30 | ДЖОНСОН ЭНД ДЖОНСОН ФАРМАСЬЮТИКАЛ РИСЕРЧ ЭНД ДИВЕЛОПМЕНТ, Эл. Эл. Си. | MODULATION OF THE GLUCOCORTICOID RECEPTOR EXPRESSION |
EP2096170B1 (en) | 2005-09-19 | 2011-08-10 | Isis Pharmaceuticals, Inc. | Modulation of glucagon receptor expression |
EP2189522A1 (en) | 2005-10-14 | 2010-05-26 | MUSC Foundation For Research Development | Targeting PAX2 for the induction of DEFB1-mediated tumor immunity and cancer therapy |
US8080534B2 (en) | 2005-10-14 | 2011-12-20 | Phigenix, Inc | Targeting PAX2 for the treatment of breast cancer |
EP2325315B1 (en) | 2005-10-28 | 2014-05-07 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of huntingtin gene |
WO2007056331A2 (en) | 2005-11-09 | 2007-05-18 | Alnylam Pharmaceuticals, Inc. | Compositions and methods for inhibiting expression of factor v leiden mutant gene |
EP1966377A2 (en) | 2005-11-21 | 2008-09-10 | Isis Pharmaceuticals, Inc. | Modulation of eif4e-bp2 expression |
JP2009524411A (en) * | 2005-12-21 | 2009-07-02 | イェール ユニバーシティー | Methods and compositions related to the regulation of riboswitches |
US8288354B2 (en) | 2005-12-28 | 2012-10-16 | The Scripps Research Institute | Natural antisense and non-coding RNA transcripts as drug targets |
JP2009522281A (en) | 2005-12-28 | 2009-06-11 | トランスレーショナル セラピューティクス,インク. | Treatment based on translational dysfunction |
JP5213723B2 (en) | 2006-01-27 | 2013-06-19 | アイシス ファーマシューティカルズ, インコーポレーテッド | Oligomer compounds and compositions for use in modulating microRNA |
US7569686B1 (en) | 2006-01-27 | 2009-08-04 | Isis Pharmaceuticals, Inc. | Compounds and methods for synthesis of bicyclic nucleic acid analogs |
JP5342881B2 (en) | 2006-01-27 | 2013-11-13 | アイシス ファーマシューティカルズ, インコーポレーテッド | 6-modified bicyclic nucleic acid analogues |
WO2007125173A2 (en) | 2006-05-03 | 2007-11-08 | Baltic Technology Development, Ltd. | Antisense agents combining strongly bound base - modified oligonucleotide and artificial nuclease |
WO2007134014A2 (en) * | 2006-05-05 | 2007-11-22 | Isis Pharmaceuticals, Inc. | Compounds and methods for modulating expression of gcgr |
US7666854B2 (en) * | 2006-05-11 | 2010-02-23 | Isis Pharmaceuticals, Inc. | Bis-modified bicyclic nucleic acid analogs |
ES2389737T3 (en) * | 2006-05-11 | 2012-10-31 | Isis Pharmaceuticals, Inc. | 5 'modified bicyclic nucleic acid analogs |
WO2007137301A2 (en) * | 2006-05-23 | 2007-11-29 | Isis Pharmaceuticals, Inc. | Modulation of chrebp expression |
WO2008097328A2 (en) * | 2006-06-23 | 2008-08-14 | Northwestern University | Asymmetric functionalized nanoparticles and methods of use |
US8198253B2 (en) | 2006-07-19 | 2012-06-12 | Isis Pharmaceuticals, Inc. | Compositions and their uses directed to HBXIP |
EP2061799A4 (en) * | 2006-09-11 | 2010-12-22 | Univ Yale | Methods and compositions for the use of lysine riboswitches |
WO2008042156A1 (en) * | 2006-09-28 | 2008-04-10 | Northwestern University | Maximizing oligonucleotide loading on gold nanoparticle |
CA2665536C (en) | 2006-10-05 | 2016-02-16 | Massachusetts Institute Of Technology | Multifunctional encoded particles for high-throughput analysis |
EP2104516B1 (en) | 2006-11-01 | 2015-01-07 | University of Rochester | Methods and compositions related to the structure and function of apobec3g |
CA2672297A1 (en) | 2006-12-11 | 2008-06-19 | University Of Utah Research Foundation | Compositions and methods for treating pathologic angiogenesis and vascular permeability |
EP2913341A1 (en) | 2006-12-22 | 2015-09-02 | University of Utah Research Foundation | Method of detecting ocular diseases and pathologic conditions and treatment of same |
US20080293053A1 (en) * | 2006-12-28 | 2008-11-27 | The Regents Of The University Of Michigan | shRNA Materials and Methods of Using Same for Inhibition of DKK-1 |
US20100093836A1 (en) | 2007-01-29 | 2010-04-15 | Isis Pharmaceuticals, Inc | Compounds and methods for modulating protein expression |
CA2691066C (en) | 2007-02-09 | 2018-07-31 | Northwestern University | Particles for detecting intracellular targets |
AU2008227458A1 (en) | 2007-03-22 | 2008-09-25 | Yale University | Methods and compositions related to riboswitches that control alternative splicing |
WO2008156987A2 (en) | 2007-05-29 | 2008-12-24 | Yale University | Riboswitches and methods and compositions for use of and with riboswitches |
US20100221821A1 (en) * | 2007-05-29 | 2010-09-02 | Yale University | Methods and compositions related to riboswitches that control alternative splicing and rna processing |
CA2688321A1 (en) | 2007-05-30 | 2008-12-11 | Isis Pharmaceuticals, Inc. | N-substituted-aminomethylene bridged bicyclic nucleic acid analogs |
EP2826863B1 (en) | 2007-05-30 | 2017-08-23 | Northwestern University | Nucleic acid functionalized nanoparticles for therapeutic applications |
US7807372B2 (en) * | 2007-06-04 | 2010-10-05 | Northwestern University | Screening sequence selectivity of oligonucleotide-binding molecules using nanoparticle based colorimetric assay |
WO2008154401A2 (en) | 2007-06-08 | 2008-12-18 | Isis Pharmaceuticals, Inc. | Carbocyclic bicyclic nucleic acid analogs |
US8278283B2 (en) * | 2007-07-05 | 2012-10-02 | Isis Pharmaceuticals, Inc. | 6-disubstituted or unsaturated bicyclic nucleic acid analogs |
AU2008286771B2 (en) | 2007-08-15 | 2013-08-15 | Isis Pharmaceuticals, Inc. | Tetrahydropyran nucleic acid analogs |
WO2009032702A2 (en) | 2007-08-28 | 2009-03-12 | Uab Research Foundation | Synthetic apolipoprotein e mimicking polypeptides and methods of use |
WO2009032693A2 (en) | 2007-08-28 | 2009-03-12 | Uab Research Foundation | Synthetic apolipoprotein e mimicking polypeptides and methods of use |
US8445217B2 (en) | 2007-09-20 | 2013-05-21 | Vanderbilt University | Free solution measurement of molecular interactions by backscattering interferometry |
WO2009039442A1 (en) * | 2007-09-21 | 2009-03-26 | California Institute Of Technology | Nfia in glial fate determination, glioma therapy and astrocytoma treatment |
MX2010003465A (en) | 2007-10-02 | 2010-07-05 | Amgen Inc | Increasing erythropoietin using nucleic acids hybridizable to micro-rna and precursors thereof. |
USRE47320E1 (en) | 2007-11-20 | 2019-03-26 | Ionis Pharmaceuticals, Inc. | Modulation of CD40 expression |
CA2708173C (en) | 2007-12-04 | 2016-02-02 | Alnylam Pharmaceuticals, Inc. | Targeting lipids |
EP3100718B1 (en) | 2008-01-02 | 2019-11-27 | Arbutus Biopharma Corporation | Improved compositions and methods for the delivery of nucleic acids |
EP2265627A2 (en) * | 2008-02-07 | 2010-12-29 | Isis Pharmaceuticals, Inc. | Bicyclic cyclohexitol nucleic acid analogs |
WO2009117589A1 (en) * | 2008-03-21 | 2009-09-24 | Isis Pharmaceuticals, Inc. | Oligomeric compounds comprising tricyclic nucleosides and methods for their use |
WO2009124295A2 (en) * | 2008-04-04 | 2009-10-08 | Isis Pharmaceuticals, Inc. | Oligomeric compounds comprising bicyclic nucleosides and having reduced toxicity |
EP2285819B1 (en) * | 2008-04-04 | 2013-10-16 | Isis Pharmaceuticals, Inc. | Oligomeric compounds comprising neutrally linked terminal bicyclic nucleosides |
CA2721183C (en) | 2008-04-11 | 2019-07-16 | Alnylam Pharmaceuticals, Inc. | Site-specific delivery of nucleic acids by combining targeting ligands with endosomolytic components |
MX2010011508A (en) | 2008-04-18 | 2011-05-03 | Baxter Int | Microsphere-based composition for preventing and/or reversing new-onset autoimmune diabetes. |
EP2280995A2 (en) * | 2008-04-29 | 2011-02-09 | Wyeth LLC | Methods for treating inflammation |
EP3081648A1 (en) | 2008-08-25 | 2016-10-19 | Excaliard Pharmaceuticals, Inc. | Antisense oligonucleotides directed against connective tissue growth factor and uses thereof |
WO2011028218A1 (en) | 2009-09-02 | 2011-03-10 | Alnylam Pharmaceuticals, Inc. | Process for triphosphate oligonucleotide synthesis |
EP2331690B1 (en) | 2008-09-02 | 2016-01-13 | Alnylam Pharmaceuticals Inc. | Compositions and methods for inhibiting expression of mutant egfr gene |
EP3587434A1 (en) | 2008-09-23 | 2020-01-01 | Alnylam Pharmaceuticals Inc. | Chemical modifications of monomers and oligonucleotides with click components for conjugation with ligands |
US8501805B2 (en) * | 2008-09-24 | 2013-08-06 | Isis Pharmaceuticals, Inc. | Substituted alpha-L-bicyclic nucleosides |
US8604192B2 (en) * | 2008-09-24 | 2013-12-10 | Isis Pharmaceuticals, Inc. | Cyclohexenyl nucleic acids analogs |
ES2475065T3 (en) | 2008-10-09 | 2014-07-10 | Tekmira Pharmaceuticals Corporation | Enhanced amino acids and methods for nucleic acid administration |
AU2009305636A1 (en) | 2008-10-15 | 2010-04-22 | Ionis Pharmaceuticals, Inc. | Modulation of Factor 11 expression |
AU2009308217B2 (en) | 2008-10-24 | 2016-01-21 | Ionis Pharmaceuticals, Inc. | 5' and 2' bis-substituted nucleosides and oligomeric compounds prepared therefrom |
WO2010048585A2 (en) | 2008-10-24 | 2010-04-29 | Isis Pharmaceuticals, Inc. | Oligomeric compounds and methods |
KR101967417B1 (en) | 2008-11-10 | 2019-04-10 | 알닐람 파마슈티칼스 인코포레이티드 | Novel lipids and compositions for the delivery of therapeutics |
CA2744207C (en) | 2008-11-24 | 2019-05-28 | Northwestern University | Polyvalent rna-nanoparticle compositions |
EP2370451B1 (en) | 2008-12-02 | 2016-11-16 | Wave Life Sciences Japan, Inc. | Method for the synthesis of phosphorus atom modified nucleic acids |
CA2745811C (en) | 2008-12-04 | 2021-07-13 | Joseph Collard | Treatment of tumor suppressor gene related diseases by inhibition of natural antisense transcript to the gene |
KR101749352B1 (en) | 2008-12-04 | 2017-06-20 | 큐알엔에이, 인크. | Treatment of sirtuin 1(sirt1) related diseases by inhibition of natural antisense transcript to sirtuin 1 |
CN102317458B (en) | 2008-12-04 | 2018-01-02 | 库尔纳公司 | Pass through treatment of the suppression of erythropoietin(EPO) (EPO) natural antisense transcript to EPO relevant diseases |
US20100233270A1 (en) | 2009-01-08 | 2010-09-16 | Northwestern University | Delivery of Oligonucleotide-Functionalized Nanoparticles |
KR101546673B1 (en) * | 2009-01-15 | 2015-08-25 | 삼성전자주식회사 | Toner for electrophotographic and process for preparing the same |
CA2750820A1 (en) | 2009-01-27 | 2010-08-05 | Qiagen Gaithersburg | Thermophilic helicase dependent amplification technology with endpoint homogenous fluorescent detection |
AU2010208035B2 (en) | 2009-01-29 | 2016-06-23 | Arbutus Biopharma Corporation | Improved lipid formulation for the delivery of nucleic acids |
US8536320B2 (en) | 2009-02-06 | 2013-09-17 | Isis Pharmaceuticals, Inc. | Tetrahydropyran nucleic acid analogs |
WO2010091308A2 (en) | 2009-02-06 | 2010-08-12 | Isis Pharmaceuticals, Inc. | Oligomeric compounds and methods |
KR101805199B1 (en) | 2009-02-12 | 2017-12-05 | 큐알엔에이, 인크. | Treatment of glial cell derived neurotrophic factor (gdnf) related diseases by inhibition of natural antisense transcript to gdnf |
EP3009150B1 (en) | 2009-02-12 | 2019-11-13 | CuRNA, Inc. | Treatment of brain derived neurotrophic factor (bdnf) related diseases by inhibition of natural antisense transcript to bdnf |
EP3424939A1 (en) | 2009-03-02 | 2019-01-09 | Alnylam Pharmaceuticals Inc. | Nucleic acid chemical modifications |
US20110319317A1 (en) | 2009-03-04 | 2011-12-29 | Opko Curna, Llc | Treatment of sirtuin 1 (sirt1) related diseases by inhibition of natural antisense transcript to sirt1 |
WO2010107733A2 (en) | 2009-03-16 | 2010-09-23 | Curna, Inc. | Treatment of nuclear factor (erythroid-derived 2)-like 2 (nrf2) related diseases by inhibition of natural antisense transcript to nrf2 |
EP2408920B1 (en) | 2009-03-17 | 2017-03-08 | CuRNA, Inc. | Treatment of delta-like 1 homolog (dlk1) related diseases by inhibition of natural antisense transcript to dlk1 |
KR20120022938A (en) | 2009-04-15 | 2012-03-12 | 노오쓰웨스턴 유니버시티 | Delivery of oligonucleotide-functionalized nanoparticles |
EP3248618A1 (en) | 2009-04-22 | 2017-11-29 | Massachusetts Institute Of Technology | Innate immune suppression enables repeated delivery of long rna molecules |
ES2661787T3 (en) | 2009-05-01 | 2018-04-04 | Curna, Inc. | Treatment of hemoglobin-related diseases (hbf / hbg) by inhibition of natural antisense transcript for hbf / hbg |
NZ596186A (en) | 2009-05-05 | 2014-03-28 | Alnylam Pharmaceuticals Inc | Lipid compositions |
CA3045126A1 (en) | 2009-05-05 | 2010-11-11 | Arbutus Biopharma Corporation | Methods of delivering oligonucleotides to immune cells |
KR101722541B1 (en) | 2009-05-06 | 2017-04-04 | 큐알엔에이, 인크. | Treatment of tristetraproline(ttp) related diseases by inhibition of natural antisense transcript to ttp |
CN103223177B (en) | 2009-05-06 | 2016-08-10 | 库尔纳公司 | By suppression therapy lipid transfer and the metabolic gene relevant disease of the natural antisense transcript for lipid transfer and metabolic gene |
WO2010132665A1 (en) | 2009-05-15 | 2010-11-18 | Yale University | Gemm riboswitches, structure-based compound design with gemm riboswitches, and methods and compositions for use of and with gemm riboswitches |
CN102575251B (en) | 2009-05-18 | 2018-12-04 | 库尔纳公司 | The relevant disease of the reprogramming factor is treated by inhibiting the natural antisense transcript for the reprogramming factor |
KR101703695B1 (en) | 2009-05-22 | 2017-02-08 | 큐알엔에이, 인크. | Treatment of transcription factor e3 (tfe3) and insulin receptor substrate 2 (irs2) related diseases by inhibition of natural antisense transcript to tfe3 |
EP2435571B1 (en) | 2009-05-28 | 2016-12-14 | CuRNA, Inc. | Treatment of antiviral gene related diseases by inhibition of natural antisense transcript to an antiviral gene |
KR101766408B1 (en) | 2009-06-10 | 2017-08-10 | 알닐람 파마슈티칼스 인코포레이티드 | Improved lipid formulation |
JP5944311B2 (en) | 2009-06-16 | 2016-07-05 | クルナ・インコーポレーテッド | Treatment of collagen gene-related diseases by suppression of natural antisense transcripts against collagen genes |
EP2443238B1 (en) | 2009-06-16 | 2017-03-22 | CuRNA, Inc. | Treatment of paraoxonase 1 (pon1) related diseases by inhibition of natural antisense transcript to pon1 |
CN102597238B (en) | 2009-06-24 | 2016-06-29 | 库尔纳公司 | The relevant disease of TNFR2 is treated by suppressing for the natural antisense transcript of tumor necrosis factor receptor 2 (TNFR2) |
WO2010151674A2 (en) | 2009-06-26 | 2010-12-29 | Curna, Inc. | Treatment of down syndrome gene related diseases by inhibition of natural antisense transcript to a down syndrome gene |
SG177564A1 (en) | 2009-07-06 | 2012-02-28 | Ontorii Inc | Novel nucleic acid prodrugs and methods of use thereof |
JP6128848B2 (en) | 2009-08-05 | 2017-05-17 | クルナ・インコーポレーテッド | Treatment of insulin gene (INS) -related diseases by suppression of natural antisense transcripts against the insulin gene (INS) |
WO2011017521A2 (en) | 2009-08-06 | 2011-02-10 | Isis Pharmaceuticals, Inc. | Bicyclic cyclohexose nucleic acid analogs |
WO2011022420A1 (en) | 2009-08-17 | 2011-02-24 | Yale University | Methylation biomarkers and methods of use |
WO2011031482A2 (en) | 2009-08-25 | 2011-03-17 | Curna, Inc. | Treatment of 'iq motif containing gtpase activating protein' (iqgap) related diseases by inhibition of natural antisense transcript to iqgap |
CA2772715C (en) | 2009-09-02 | 2019-03-26 | Genentech, Inc. | Mutant smoothened and methods of using the same |
CN102711826B (en) | 2009-10-22 | 2017-03-29 | 霍夫曼-拉罗奇有限公司 | For the method and composition that the HEPSIN for regulating and controlling macrophage-stimulating albumen is activated |
WO2011056687A2 (en) | 2009-10-27 | 2011-05-12 | Swift Biosciences, Inc. | Polynucleotide primers and probes |
CA2779099C (en) | 2009-10-30 | 2021-08-10 | Northwestern University | Templated nanoconjugates |
US20130084565A1 (en) | 2009-11-03 | 2013-04-04 | University Of Virginia Patent Foundation | Versatile, visible method for detecting polymeric analytes |
AU2010321576A1 (en) | 2009-11-23 | 2012-06-07 | Swift Biosciences, Inc. | Devices to extend single stranded target molecules |
CA2781887C (en) | 2009-11-30 | 2018-03-27 | Genentech, Inc. | Compositions and methods for the diagnosis and treatment of tumor |
CA3044884A1 (en) | 2009-12-07 | 2011-06-16 | Arbutus Biopharma Corporation | Compositions for nucleic acid delivery |
WO2011084455A2 (en) | 2009-12-16 | 2011-07-14 | Opko Curna, Llc. | Treatment of membrane bound transcription factor peptidase, site 1 (mbtps1) related diseases by inhibition of natural antisense transcript to mbtps1 |
ES2749426T3 (en) | 2009-12-18 | 2020-03-20 | Univ British Columbia | Nucleic Acid Administration Methods and Compositions |
KR101793753B1 (en) | 2009-12-23 | 2017-11-03 | 큐알엔에이, 인크. | Treatment of uncoupling protein 2 (ucp2) related diseases by inhibition of natural antisense transcript to ucp2 |
CN102869776B (en) | 2009-12-23 | 2017-06-23 | 库尔纳公司 | HGF relevant diseases are treated by suppressing the natural antisense transcript of HGF (HGF) |
EP2519633B1 (en) | 2009-12-29 | 2017-10-25 | CuRNA, Inc. | Treatment of nuclear respiratory factor 1 (nrf1) related diseases by inhibition of natural antisense transcript to nrf1 |
WO2011090741A2 (en) | 2009-12-29 | 2011-07-28 | Opko Curna, Llc | TREATMENT OF TUMOR PROTEIN 63 (p63) RELATED DISEASES BY INHIBITION OF NATURAL ANTISENSE TRANSCRIPT TO p63 |
NO2521784T3 (en) | 2010-01-04 | 2018-05-05 | ||
US8912157B2 (en) | 2010-01-06 | 2014-12-16 | Curna, Inc. | Treatment of pancreatic developmental gene related diseases by inhibition of natural antisense transcript to a pancreatic developmental gene |
US8779118B2 (en) | 2010-01-11 | 2014-07-15 | Isis Pharmaceuticals, Inc. | Base modified bicyclic nucleosides and oligomeric compounds prepared therefrom |
DK2524039T3 (en) | 2010-01-11 | 2018-03-12 | Curna Inc | TREATMENT OF GENDER HORMON-BINDING GLOBULIN (SHBG) RELATED DISEASES BY INHIBITION OF NATURAL ANTISENCE TRANSCRIPTS TO SHBG |
EP2524042A2 (en) | 2010-01-12 | 2012-11-21 | Yale University | Structured rna motifs and compounds and methods for their use |
WO2011091390A2 (en) | 2010-01-25 | 2011-07-28 | Opko Curna, Llc | Treatment of rnase h1 related diseases by inhibition of natural antisense transcript to rnase h1 |
WO2011100131A2 (en) | 2010-01-28 | 2011-08-18 | Alnylam Pharmacuticals, Inc. | Monomers and oligonucleotides comprising cycloaddition adduct(s) |
WO2011094580A2 (en) | 2010-01-28 | 2011-08-04 | Alnylam Pharmaceuticals, Inc. | Chelated copper for use in the preparation of conjugated oligonucleotides |
US20130028889A1 (en) | 2010-02-04 | 2013-01-31 | Ico Therapeutics Inc. | Dosing regimens for treating and preventing ocular disorders using c-raf antisense |
WO2011103528A2 (en) | 2010-02-22 | 2011-08-25 | Opko Curna Llc | Treatment of pyrroline-5-carboxylate reductase 1 (pycr1) related diseases by inhibition of natural antisense transcript to pycr1 |
WO2011105900A2 (en) | 2010-02-23 | 2011-09-01 | Academisch Ziekenhuis Bij De Universiteit Van Amsterdam | Antagonists of complement component 8-alpha (c8-alpha) and uses thereof |
WO2011105901A2 (en) | 2010-02-23 | 2011-09-01 | Academisch Ziekenhuis Bij De Universiteit Van Amsterdam | Antagonists of complement component 9 (c9) and uses thereof |
WO2011105902A2 (en) | 2010-02-23 | 2011-09-01 | Academisch Ziekenhuis Bij De Universiteit Van Amsterdam | Antagonists of complement component 8-beta (c8-beta) and uses thereof |
EP2538981B1 (en) | 2010-02-23 | 2017-12-20 | F. Hoffmann-La Roche AG | Compositions and methods for the diagnosis and treatment of tumor |
WO2011112516A1 (en) | 2010-03-08 | 2011-09-15 | Ico Therapeutics Inc. | Treating and preventing hepatitis c virus infection using c-raf kinase antisense oligonucleotides |
US20130101512A1 (en) | 2010-03-12 | 2013-04-25 | Chad A. Mirkin | Crosslinked polynucleotide structure |
WO2011112732A2 (en) | 2010-03-12 | 2011-09-15 | The Brigham And Women's Hospital, Inc. | Methods of treating vascular inflammatory disorders |
WO2011115817A1 (en) | 2010-03-16 | 2011-09-22 | Isis Pharmaceuticals, Inc. | Methods of preparing 2'-o-substituted purine nucleosides |
US9193752B2 (en) | 2010-03-17 | 2015-11-24 | Isis Pharmaceuticals, Inc. | 5′-substituted bicyclic nucleosides and oligomeric compounds prepared therefrom |
US8889350B2 (en) | 2010-03-26 | 2014-11-18 | Swift Biosciences, Inc. | Methods and compositions for isolating polynucleotides |
ES2893199T3 (en) | 2010-03-29 | 2022-02-08 | Alnylam Pharmaceuticals Inc | dsRNA therapy for transthyretin (TTR)-related ocular amyloidosis |
US9102938B2 (en) | 2010-04-01 | 2015-08-11 | Alnylam Pharmaceuticals, Inc. | 2′ and 5′ modified monomers and oligonucleotides |
US9044494B2 (en) | 2010-04-09 | 2015-06-02 | Curna, Inc. | Treatment of fibroblast growth factor 21 (FGF21) related diseases by inhibition of natural antisense transcript to FGF21 |
WO2011133695A2 (en) | 2010-04-20 | 2011-10-27 | Swift Biosciences, Inc. | Materials and methods for nucleic acid fractionation by solid phase entrapment and enzyme-mediated detachment |
WO2011133871A2 (en) | 2010-04-22 | 2011-10-27 | Alnylam Pharmaceuticals, Inc. | 5'-end derivatives |
US10913767B2 (en) | 2010-04-22 | 2021-02-09 | Alnylam Pharmaceuticals, Inc. | Oligonucleotides comprising acyclic and abasic nucleosides and analogs |
US20130260460A1 (en) | 2010-04-22 | 2013-10-03 | Isis Pharmaceuticals Inc | Conformationally restricted dinucleotide monomers and oligonucleotides |
EP2625186B1 (en) | 2010-04-28 | 2016-07-27 | Ionis Pharmaceuticals, Inc. | 5' modified nucleosides and oligomeric compounds prepared therefrom |
WO2011139695A2 (en) | 2010-04-28 | 2011-11-10 | Isis Pharmaceuticals, Inc. | Modified 5' diphosphate nucleosides and oligomeric compounds prepared therefrom |
KR101869570B1 (en) | 2010-04-28 | 2018-06-20 | 아이오니스 파마수티컬즈, 인코포레이티드 | Modified nucleosides and oligomeric compounds prepared therefrom |
WO2011139911A2 (en) | 2010-04-29 | 2011-11-10 | Isis Pharmaceuticals, Inc. | Lipid formulated single stranded rna |
WO2011139917A1 (en) | 2010-04-29 | 2011-11-10 | Isis Pharmaceuticals, Inc. | Modulation of transthyretin expression |
MA34291B1 (en) | 2010-05-03 | 2013-06-01 | Genentech Inc | COMPOSITIONS AND METHODS FOR DIAGNOSING AND TREATING A TUMOR |
EP2566966A4 (en) | 2010-05-03 | 2013-12-11 | Curna Inc | Treatment of sirtuin (sirt) related diseases by inhibition of natural antisense transcript to a sirtuin (sirt) |
TWI586356B (en) | 2010-05-14 | 2017-06-11 | 可娜公司 | Treatment of par4 related diseases by inhibition of natural antisense transcript to par4 |
WO2011150227A1 (en) | 2010-05-26 | 2011-12-01 | Qiagen Gaithersburg, Inc. | Quantitative helicase assay |
CA2799207C (en) | 2010-05-26 | 2019-03-26 | Curna, Inc. | Treatment of atonal homolog 1 (atoh1) related diseases by inhibition of natural antisense transcript to atoh1 |
US20130203045A1 (en) | 2010-05-26 | 2013-08-08 | University Of Virginia Patent Foundation | Method for detecting nucleic acids based on aggregate formation |
CA2802049C (en) | 2010-06-07 | 2018-07-10 | Firefly Bioworks, Inc. | Scanning multifunctional particles |
WO2011156278A1 (en) | 2010-06-07 | 2011-12-15 | Isis Pharmaceuticals, Inc. | Bicyclic nucleosides and oligomeric compounds prepared therefrom |
EP2580228B1 (en) | 2010-06-08 | 2016-03-23 | Ionis Pharmaceuticals, Inc. | Substituted 2'-amino and 2'-thio-bicyclic nucleosides and oligomeric compounds prepared therefrom |
US9638632B2 (en) | 2010-06-11 | 2017-05-02 | Vanderbilt University | Multiplexed interferometric detection system and method |
WO2011159836A2 (en) | 2010-06-15 | 2011-12-22 | Isis Pharmaceuticals, Inc. | Compounds and methods for modulating interaction between proteins and target nucleic acids |
WO2011163466A1 (en) | 2010-06-23 | 2011-12-29 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Regulation of skin pigmentation by neuregulin-1 (nrg-1) |
US8980860B2 (en) | 2010-07-14 | 2015-03-17 | Curna, Inc. | Treatment of discs large homolog (DLG) related diseases by inhibition of natural antisense transcript to DLG |
WO2012012467A2 (en) | 2010-07-19 | 2012-01-26 | Isis Pharmaceuticals, Inc. | Modulation of nuclear-retained rna |
WO2012016184A2 (en) | 2010-07-30 | 2012-02-02 | Alnylam Pharmaceuticals, Inc. | Methods and compositions for delivery of active agents |
WO2012016188A2 (en) | 2010-07-30 | 2012-02-02 | Alnylam Pharmaceuticals, Inc. | Methods and compositions for delivery of active agents |
US20130143955A1 (en) | 2010-08-09 | 2013-06-06 | Yale University | Cyclic di-GMP-II Riboswitches, Motifs, and Compounds, and Methods for Their Use |
JP5868324B2 (en) | 2010-09-24 | 2016-02-24 | 株式会社Wave Life Sciences Japan | Asymmetric auxiliary group |
US8481680B2 (en) | 2010-10-05 | 2013-07-09 | Genentech, Inc. | Mutant smoothened and methods of using the same |
US8993533B2 (en) | 2010-10-06 | 2015-03-31 | Curna, Inc. | Treatment of sialidase 4 (NEU4) related diseases by inhibition of natural antisense transcript to NEU4 |
US20140031250A1 (en) | 2010-10-07 | 2014-01-30 | David Tsai Ting | Biomarkers of Cancer |
US8648053B2 (en) | 2010-10-20 | 2014-02-11 | Rosalind Franklin University Of Medicine And Science | Antisense oligonucleotides that target a cryptic splice site in Ush1c as a therapeutic for Usher syndrome |
CA2815212A1 (en) | 2010-10-22 | 2012-04-26 | Curna, Inc. | Treatment of alpha-l-iduronidase (idua) related diseases by inhibition of natural antisense transcript to idua |
CN103201387B (en) | 2010-10-27 | 2018-02-02 | 库尔纳公司 | IFRD1 relevant diseases are treated by suppressing the natural antisense transcript of interferon correlative development regulatory factor 1 (IFRD1) |
CN110123830A (en) | 2010-11-09 | 2019-08-16 | 阿尔尼拉姆医药品有限公司 | Composition and method for inhibiting the lipid of the expression of Eg5 and VEGF gene to prepare |
EP3260540A1 (en) | 2010-11-12 | 2017-12-27 | The General Hospital Corporation | Polycomb-associated non-coding rnas |
EP2640853B1 (en) | 2010-11-17 | 2018-12-26 | Ionis Pharmaceuticals, Inc. | Modulation of alpha synuclein expression |
WO2012071238A2 (en) | 2010-11-23 | 2012-05-31 | Opko Curna Llc | Treatment of nanog related diseases by inhibition of natural antisense transcript to nanog |
US9150926B2 (en) | 2010-12-06 | 2015-10-06 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Diagnosis and treatment of adrenocortical tumors using human microRNA-483 |
DK3202760T3 (en) | 2011-01-11 | 2019-11-25 | Alnylam Pharmaceuticals Inc | PEGYLED LIPIDS AND THEIR USE FOR PHARMACEUTICAL SUPPLY |
WO2012097261A2 (en) | 2011-01-14 | 2012-07-19 | The General Hospital Corporation | Methods targeting mir-128 for regulating cholesterol/lipid metabolism |
EP2670404B1 (en) | 2011-02-02 | 2018-08-29 | The Trustees of Princeton University | Sirtuin modulators as virus production modulators |
SI2670411T1 (en) | 2011-02-02 | 2019-06-28 | Excaliard Pharmaceuticals, Inc. | Antisense compounds targeting connective tissue growth factor (ctgf) for use in a method of treating keloids or hypertrophic scars |
WO2012109157A2 (en) | 2011-02-07 | 2012-08-16 | The Governing Council Of The University Of Toronto | Bioprobes and methods of use thereof |
EP3467109A1 (en) | 2011-02-08 | 2019-04-10 | Ionis Pharmaceuticals, Inc. | Oligomeric compounds comprising bicyclic nucleotides and uses thereof |
US9562853B2 (en) | 2011-02-22 | 2017-02-07 | Vanderbilt University | Nonaqueous backscattering interferometric methods |
WO2012138453A1 (en) | 2011-04-03 | 2012-10-11 | The General Hospital Corporation | Efficient protein expression in vivo using modified rna (mod-rna) |
US20140186844A1 (en) | 2011-04-26 | 2014-07-03 | Swift Biosciences, Inc. | Polynucleotide primers and probes |
WO2012151324A1 (en) | 2011-05-02 | 2012-11-08 | Isis Pharmaceuticals, Inc. | Antisense compounds targeting genes associated with usher syndrome |
WO2012151268A1 (en) | 2011-05-02 | 2012-11-08 | University Of Virginia Patent Foundation | Method and system for high throughput optical and label free detection of analytes |
WO2012151289A2 (en) | 2011-05-02 | 2012-11-08 | University Of Virginia Patent Foundation | Method and system to detect aggregate formation on a substrate |
WO2012170347A1 (en) | 2011-06-09 | 2012-12-13 | Isis Pharmaceuticals, Inc. | Bicyclic nucleosides and oligomeric compounds prepared therefrom |
RU2620980C2 (en) | 2011-06-09 | 2017-05-30 | Курна, Инк. | Treatment of diseases associated with frataxin (fxn), by inhibiting natural antisense fxn transcript |
US9222093B2 (en) | 2011-06-30 | 2015-12-29 | The University Of Hong Kong | Two-way, portable riboswitch mediated gene expression control device |
US20140227293A1 (en) | 2011-06-30 | 2014-08-14 | Trustees Of Boston University | Method for controlling tumor growth, angiogenesis and metastasis using immunoglobulin containing and proline rich receptor-1 (igpr-1) |
RU2014105311A (en) | 2011-07-19 | 2015-08-27 | Уэйв Лайф Сайенсес Пте. Лтд. | METHODS FOR SYNTHESIS OF FUNCTIONALIZED NUCLEIC ACIDS |
EP2742136B1 (en) | 2011-08-11 | 2017-09-27 | Ionis Pharmaceuticals, Inc. | Gapped oligomeric compounds comprising 5'-modified deoxyribonucleosides in the gap and uses thereof |
DK2751270T3 (en) | 2011-08-29 | 2018-10-29 | Ionis Pharmaceuticals Inc | OLIGOMER-CONJUGATE COMPLEXES AND THEIR USE |
EP2751269B1 (en) | 2011-08-29 | 2016-03-23 | Ionis Pharmaceuticals, Inc. | Methods and compounds useful in conditions related to repeat expansion |
EP2756080B1 (en) | 2011-09-14 | 2019-02-20 | Translate Bio MA, Inc. | Multimeric oligonucleotide compounds |
EP2755692B1 (en) | 2011-09-14 | 2020-11-25 | Northwestern University | Nanoconjugates able to cross the blood-brain barrier |
US9580713B2 (en) | 2011-09-17 | 2017-02-28 | Yale University | Fluoride-responsive riboswitches, fluoride transporters, and methods of use |
CA2849476A1 (en) | 2011-09-27 | 2013-04-04 | Alnylam Pharmaceuticals, Inc. | Di-aliphatic substituted pegylated lipids |
EP2766482B1 (en) | 2011-10-11 | 2016-12-07 | The Brigham and Women's Hospital, Inc. | Micrornas in neurodegenerative disorders |
ES2687951T3 (en) | 2011-10-14 | 2018-10-30 | F. Hoffmann-La Roche Ag | Anti-HtrA1 antibodies and procedures for use |
US9243291B1 (en) | 2011-12-01 | 2016-01-26 | Isis Pharmaceuticals, Inc. | Methods of predicting toxicity |
EP2790736B1 (en) | 2011-12-12 | 2018-01-31 | Oncoimmunin, Inc. | In vivo delivery of oligonucleotides |
WO2013106770A1 (en) | 2012-01-11 | 2013-07-18 | Isis Pharmaceuticals, Inc. | Compositions and methods for modulation of ikbkap splicing |
SG11201405669XA (en) | 2012-03-13 | 2014-10-30 | Swift Biosciences Inc | Methods and compositions for size-controlled homopolymer tailing of substrate polynucleotides by a nucleic acid polymerase |
EP2639238A1 (en) | 2012-03-15 | 2013-09-18 | Universität Bern | Tricyclic nucleosides and oligomeric compounds prepared therefrom |
JP2015511494A (en) | 2012-03-15 | 2015-04-20 | キュアナ,インク. | Treatment of BDNF-related diseases by inhibition of natural antisense transcripts against brain-derived neurotrophic factor (BDNF) |
WO2013138662A1 (en) | 2012-03-16 | 2013-09-19 | 4S3 Bioscience, Inc. | Antisense conjugates for decreasing expression of dmpk |
AU2013202595B2 (en) | 2012-03-30 | 2016-04-21 | Biogen Ma Inc. | Methods for modulating Tau expression for reducing seizure and modifying a neurodegenerative syndrome |
EP2850092B1 (en) | 2012-04-09 | 2017-03-01 | Ionis Pharmaceuticals, Inc. | Tricyclic nucleic acid analogs |
WO2013154799A1 (en) | 2012-04-09 | 2013-10-17 | Isis Pharmaceuticals, Inc. | Tricyclic nucleosides and oligomeric compounds prepared therefrom |
JP2015518485A (en) | 2012-04-20 | 2015-07-02 | アプタミアール セラピューティクス インコーポレイテッド | Thermogenic miRNA regulator |
EP2839006B1 (en) | 2012-04-20 | 2018-01-03 | Ionis Pharmaceuticals, Inc. | Oligomeric compounds comprising bicyclic nucleotides and uses thereof |
EP2841572B1 (en) | 2012-04-27 | 2019-06-19 | Duke University | Genetic correction of mutated genes |
US9273949B2 (en) | 2012-05-11 | 2016-03-01 | Vanderbilt University | Backscattering interferometric methods |
AU2013262663A1 (en) | 2012-05-16 | 2015-01-22 | The General Hospital Corporation D/B/A Massachusetts General Hospital | Compositions and methods for modulating gene expression |
EA201492123A1 (en) | 2012-05-16 | 2015-10-30 | Рана Терапьютикс, Инк. | COMPOSITIONS AND METHODS FOR MODULATING THE EXPRESSION OF THE SMN GENES FAMILY |
US9574193B2 (en) | 2012-05-17 | 2017-02-21 | Ionis Pharmaceuticals, Inc. | Methods and compositions for modulating apolipoprotein (a) expression |
US20160002624A1 (en) | 2012-05-17 | 2016-01-07 | Isis Pharmaceuticals, Inc. | Antisense oligonucleotide compositions |
US9828602B2 (en) | 2012-06-01 | 2017-11-28 | Ionis Pharmaceuticals, Inc. | Antisense compounds targeting genes associated with fibronectin |
US9487780B2 (en) | 2012-06-01 | 2016-11-08 | Ionis Pharmaceuticals, Inc. | Antisense compounds targeting genes associated with fibronectin |
WO2013184209A1 (en) | 2012-06-04 | 2013-12-12 | Ludwig Institute For Cancer Research Ltd. | Mif for use in methods of treating subjects with a neurodegenerative disorder |
WO2013185097A1 (en) | 2012-06-08 | 2013-12-12 | The Regents Of The University Of Michigan | Ultrasound-triggerable agents for tissue engineering |
SG11201500243WA (en) | 2012-07-13 | 2015-04-29 | Shin Nippon Biomedical Lab Ltd | Chiral nucleic acid adjuvant |
SG11201500239VA (en) | 2012-07-13 | 2015-03-30 | Wave Life Sciences Japan | Asymmetric auxiliary group |
US9982257B2 (en) | 2012-07-13 | 2018-05-29 | Wave Life Sciences Ltd. | Chiral control |
US20140038182A1 (en) | 2012-07-17 | 2014-02-06 | Dna Logix, Inc. | Cooperative primers, probes, and applications thereof |
WO2014022852A1 (en) | 2012-08-03 | 2014-02-06 | Aptamir Therapeutics, Inc. | Cell-specific delivery of mirna modulators for the treatment of obesity and related disorders |
CN104736551B (en) | 2012-08-15 | 2017-07-28 | Ionis制药公司 | The method for preparing oligomeric compounds using improved end-blocking scheme |
CA2884245C (en) | 2012-09-06 | 2023-03-14 | The University Of Chicago | Antisense polynucleotides to induce exon skipping and methods of treating dystrophies |
US9695418B2 (en) | 2012-10-11 | 2017-07-04 | Ionis Pharmaceuticals, Inc. | Oligomeric compounds comprising bicyclic nucleosides and uses thereof |
EP4144845A1 (en) | 2012-10-12 | 2023-03-08 | Ionis Pharmaceuticals, Inc. | Antisense compounds and uses thereof |
DK2906256T3 (en) | 2012-10-12 | 2018-11-19 | Ionis Pharmaceuticals Inc | SELECTIVE ANTISENSE COMPOUNDS AND APPLICATIONS THEREOF |
US9029335B2 (en) | 2012-10-16 | 2015-05-12 | Isis Pharmaceuticals, Inc. | Substituted 2′-thio-bicyclic nucleosides and oligomeric compounds prepared therefrom |
US9695475B2 (en) | 2012-12-11 | 2017-07-04 | Ionis Pharmaceuticals, Inc. | Competitive modulation of microRNAs |
WO2014121287A2 (en) | 2013-02-04 | 2014-08-07 | Isis Pharmaceuticals, Inc. | Selective antisense compounds and uses thereof |
KR20150130430A (en) | 2013-03-14 | 2015-11-23 | 아이시스 파마수티컬즈 인코포레이티드 | Compositions and methods for modulating tau expression |
US9347095B2 (en) | 2013-03-15 | 2016-05-24 | Bio-Rad Laboratories, Inc. | Digital assays for mutation detection |
EP2978446B1 (en) | 2013-03-27 | 2020-03-04 | The General Hospital Corporation | Anti-cd33 antibody for use in treating alzheimer's disease |
WO2015012916A2 (en) | 2013-04-23 | 2015-01-29 | Northwestern University | Metal-ligand coordination polymer nanoparticles and methods for making |
SG10201906382QA (en) | 2013-05-01 | 2019-08-27 | Ionis Pharmaceuticals Inc | Compositions and methods for modulating hbv and ttr expression |
AU2014280847B2 (en) | 2013-06-13 | 2019-07-04 | Antisense Therapeutics Ltd | Combination therapy |
CA2916252A1 (en) | 2013-06-21 | 2014-12-24 | Isis Pharmaceuticals, Inc. | Compositions and methods for modulation of target nucleic acids |
EP3022176B8 (en) | 2013-07-15 | 2019-12-25 | The Regents of the University of California | Azacyclic constrained analogs of fty720 |
TWI657819B (en) | 2013-07-19 | 2019-05-01 | 美商Ionis製藥公司 | Compositions for modulating tau expression |
US10435430B2 (en) | 2013-07-31 | 2019-10-08 | Ionis Pharmaceuticals, Inc. | Methods and compounds useful in conditions related to repeat expansion |
ES2773547T3 (en) | 2013-08-08 | 2020-07-13 | Scripps Research Inst | An in vitro nucleic acid site specific enzymatic labeling procedure by incorporating unnatural nucleotides |
TW201536329A (en) | 2013-08-09 | 2015-10-01 | Isis Pharmaceuticals Inc | Compounds and methods for modulation of dystrophia myotonica-protein kinase (DMPK) expression |
WO2015042447A1 (en) | 2013-09-20 | 2015-03-26 | Isis Pharmaceuticals, Inc. | Targeted therapeutic nucleosides and their use |
WO2015054451A1 (en) | 2013-10-09 | 2015-04-16 | The United States Of America As Represented By The Secretary Department Of Health And Human Services | Detection of hepatitis delta virus (hdv) for the diagnosis and treatment of sjögren's syndrome and lymphoma |
US11162096B2 (en) | 2013-10-14 | 2021-11-02 | Ionis Pharmaceuticals, Inc | Methods for modulating expression of C9ORF72 antisense transcript |
WO2015066708A1 (en) | 2013-11-04 | 2015-05-07 | Northwestern University | Quantification and spatio-temporal tracking of a target using a spherical nucleic acid (sna) |
US10752940B2 (en) | 2013-11-08 | 2020-08-25 | Ionis Pharmaceuticals, Inc. | Compounds and methods for detecting oligonucleotides |
ES2797679T3 (en) | 2013-12-02 | 2020-12-03 | Ionis Pharmaceuticals Inc | Antisense compounds and their uses |
WO2015126502A2 (en) | 2013-12-03 | 2015-08-27 | Northwestern University | Liposomal particles, methods of making same and uses thereof |
US10385388B2 (en) | 2013-12-06 | 2019-08-20 | Swift Biosciences, Inc. | Cleavable competitor polynucleotides |
CA2844640A1 (en) | 2013-12-06 | 2015-06-06 | The University Of British Columbia | Method for treatment of castration-resistant prostate cancer |
JPWO2015108047A1 (en) | 2014-01-15 | 2017-03-23 | 株式会社新日本科学 | Chiral nucleic acid adjuvant having immunity induction activity and immunity induction activator |
US10149905B2 (en) | 2014-01-15 | 2018-12-11 | Shin Nippon Biomedical Laboratories, Ltd. | Chiral nucleic acid adjuvant having antitumor effect and antitumor agent |
JPWO2015108046A1 (en) | 2014-01-15 | 2017-03-23 | 株式会社新日本科学 | Chiral nucleic acid adjuvant and antiallergic agent having antiallergic action |
DK3094728T3 (en) | 2014-01-16 | 2022-05-16 | Wave Life Sciences Ltd | KIRALT DESIGN |
EP3102197B1 (en) | 2014-02-04 | 2018-08-29 | Genentech, Inc. | Mutant smoothened and methods of using the same |
EP3119789B1 (en) | 2014-03-17 | 2020-04-22 | Ionis Pharmaceuticals, Inc. | Bicyclic carbocyclic nucleosides and oligomeric compounds prepared therefrom |
RU2019130898A (en) | 2014-03-19 | 2019-11-11 | Ионис Фармасьютикалз, Инк. | COMPOSITIONS FOR MODULATION OF ATAXIN 2 EXPRESSION |
US10006027B2 (en) | 2014-03-19 | 2018-06-26 | Ionis Pharmaceuticals, Inc. | Methods for modulating Ataxin 2 expression |
RU2704619C2 (en) | 2014-04-01 | 2019-10-30 | Биоген Ма Инк. | Compositions for modulating expression of sod-1 |
EP3943607A1 (en) | 2014-04-09 | 2022-01-26 | The Scripps Research Institute | Import of unnatural or modified nucleoside triphosphates into cells via nucleic acid triphosphate transporters |
US10221416B2 (en) | 2014-04-24 | 2019-03-05 | Ionis Pharmaceuticals, Inc. | Oligomeric compounds comprising alpha-beta-constrained nucleic acid |
DK3137476T3 (en) | 2014-04-28 | 2019-11-18 | Ionis Pharmaceuticals Inc | LINKER-MODIFIED OLIGOMER COMPOUNDS |
EP3137604B1 (en) | 2014-05-01 | 2020-07-15 | Ionis Pharmaceuticals, Inc. | Compositions and methods for modulating growth hormone receptor expression |
SG11201608502TA (en) | 2014-05-01 | 2016-11-29 | Ionis Pharmaceuticals Inc | Compositions and methods for modulating complement factor b expression |
SI3137605T1 (en) | 2014-05-01 | 2021-02-26 | Ionis Pharmaceuticals, Inc. | Compositions and methods for modulating angiopoietin-like 3 expression |
US10570169B2 (en) | 2014-05-22 | 2020-02-25 | Ionis Pharmaceuticals, Inc. | Conjugated antisense compounds and their use |
CN106661580B (en) | 2014-06-10 | 2022-02-15 | 鹿特丹伊拉斯谟大学医疗中心 | Antisense oligonucleotides for treating pompe disease |
EP3161159B1 (en) | 2014-06-25 | 2020-08-05 | The General Hospital Corporation | Targeting human satellite ii (hsatii) |
US9951327B1 (en) | 2014-07-17 | 2018-04-24 | Integrated Dna Technologies, Inc. | Efficient and rapid method for assembling and cloning double-stranded DNA fragments |
MX2017001432A (en) | 2014-07-31 | 2017-05-09 | Uab Res Found | Apoe mimetic peptides and higher potency to clear plasma cholesterol. |
CA2958431A1 (en) | 2014-08-19 | 2016-02-25 | Northwestern University | Protein/oligonucleotide core-shell nanoparticle therapeutics |
CN107074767A (en) | 2014-08-20 | 2017-08-18 | 西北大学 | Unlimited coordination polymer nano particle-nucleic acid conjugate of the bio-compatible adjusted for antisense gene |
WO2016033424A1 (en) | 2014-08-29 | 2016-03-03 | Genzyme Corporation | Methods for the prevention and treatment of major adverse cardiovascular events using compounds that modulate apolipoprotein b |
WO2016044271A2 (en) | 2014-09-15 | 2016-03-24 | Children's Medical Center Corporation | Methods and compositions to increase somatic cell nuclear transfer (scnt) efficiency by removing histone h3-lysine trimethylation |
EP3198012B1 (en) | 2014-09-26 | 2019-09-04 | University of Massachusetts | Rna-modulating agents |
WO2016057693A1 (en) | 2014-10-10 | 2016-04-14 | Alnylam Pharmaceuticals, Inc. | Methods and compositions for inhalation delivery of conjugated oligonucleotide |
WO2016077540A1 (en) | 2014-11-12 | 2016-05-19 | Ionis Pharmaceuticals, Inc. | Compounds and methods for the modulation of comp |
CN107106493A (en) | 2014-11-21 | 2017-08-29 | 西北大学 | The sequence-specific cellular uptake of spherical nucleic acid nano particle conjugate |
EP3229842B1 (en) | 2014-12-08 | 2022-07-06 | The Board of Regents of The University of Texas System | Lipocationic polymers and uses thereof |
JP6997623B2 (en) | 2014-12-12 | 2022-02-04 | エム. ウルフ、トッド | Compositions and Methods for Editing Intracellular Nucleic Acids Utilizing Oligonucleotides |
US9688707B2 (en) | 2014-12-30 | 2017-06-27 | Ionis Pharmaceuticals, Inc. | Bicyclic morpholino compounds and oligomeric compounds prepared therefrom |
WO2016112132A1 (en) | 2015-01-06 | 2016-07-14 | Ionis Pharmaceuticals, Inc. | Compositions for modulating expression of c9orf72 antisense transcript |
WO2016115490A1 (en) | 2015-01-16 | 2016-07-21 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulation of dux4 |
JP2018506715A (en) | 2015-01-23 | 2018-03-08 | ヴァンダービルト ユニバーシティー | Robust interferometer and method of use |
EP3256487A4 (en) | 2015-02-09 | 2018-07-18 | Duke University | Compositions and methods for epigenome editing |
EP3262173A2 (en) | 2015-02-23 | 2018-01-03 | Crispr Therapeutics AG | Materials and methods for treatment of human genetic diseases including hemoglobinopathies |
US11129844B2 (en) | 2015-03-03 | 2021-09-28 | Ionis Pharmaceuticals, Inc. | Compositions and methods for modulating MECP2 expression |
WO2016164463A1 (en) | 2015-04-07 | 2016-10-13 | The General Hospital Corporation | Methods for reactivating genes on the inactive x chromosome |
PL3283500T3 (en) | 2015-04-08 | 2021-05-31 | The University Of Chicago | Compositions and methods for correcting limb girdle muscular dystrophy type 2c using exon skipping |
WO2016167780A1 (en) | 2015-04-16 | 2016-10-20 | Ionis Pharmaceuticals, Inc. | Compositions for modulating expression of c9orf72 antisense transcript |
CN108139375A (en) | 2015-06-26 | 2018-06-08 | 贝斯以色列女执事医疗中心股份有限公司 | Target the cancer therapy for inhibiting the four transmembrane proteins 33 (TSPAN33) in cell derived from marrow sample |
EP3314027A4 (en) | 2015-06-29 | 2019-07-03 | Caris Science, Inc. | Therapeutic oligonucleotides |
EP3313989A4 (en) | 2015-06-29 | 2018-12-05 | Ionis Pharmaceuticals, Inc. | Modified crispr rna and modified single crispr rna and uses thereof |
MY192997A (en) | 2015-07-10 | 2022-09-20 | Ionis Pharmaceuticals Inc | Modulators of diacyglycerol acyltransferase 2 (dgat2) |
CA2993652A1 (en) | 2015-07-28 | 2017-02-02 | Caris Science, Inc. | Targeted oligonucleotides |
WO2017021961A1 (en) | 2015-08-04 | 2017-02-09 | Yeda Research And Development Co. Ltd. | Methods of screening for riboswitches and attenuators |
CN108271360B (en) | 2015-09-14 | 2023-01-24 | 得克萨斯州大学系统董事会 | Lipophilic cationic dendritic polymer and use thereof |
CA2999177A1 (en) | 2015-09-24 | 2017-03-30 | The Regents Of The University Of California | Synthetic sphingolipid-like molecules, drugs, methods of their synthesis and methods of treatment |
AU2016339053A1 (en) | 2015-09-24 | 2018-04-12 | Crispr Therapeutics Ag | Novel family of RNA-programmable endonucleases and their uses in genome editing and other applications |
RU2018113709A (en) | 2015-09-24 | 2019-10-30 | Айонис Фармасьютикалз, Инк. | KRAS EXPRESSION MODULATORS |
WO2017053781A1 (en) | 2015-09-25 | 2017-03-30 | Ionis Pharmaceuticals, Inc. | Compositions and methods for modulating ataxin 3 expression |
WO2017058672A1 (en) | 2015-09-29 | 2017-04-06 | The Regents Of The University Of Michigan Office Of Technology Transfer | Biodegradable hydrogel for tissue expansion |
EP4089175A1 (en) | 2015-10-13 | 2022-11-16 | Duke University | Genome engineering with type i crispr systems in eukaryotic cells |
EP3368063B1 (en) | 2015-10-28 | 2023-09-06 | Vertex Pharmaceuticals Inc. | Materials and methods for treatment of duchenne muscular dystrophy |
SI3368578T1 (en) | 2015-10-30 | 2021-08-31 | F. Hoffmann-La Roche Ag | Anti-htra1 antibodies and methods of use thereof |
WO2017075670A1 (en) | 2015-11-05 | 2017-05-11 | Children's Hospital Los Angeles | "mobilizing leukemia cells" |
BR112018008971A2 (en) | 2015-11-06 | 2018-11-27 | Crispr Therapeutics Ag | Materials and Methods for Treatment of Type 1a Glycogen Storage Disease |
US20190046555A1 (en) | 2015-11-06 | 2019-02-14 | Ionis Pharmaceuticals, Inc. | Conjugated antisense compounds for use in therapy |
PE20181180A1 (en) | 2015-11-06 | 2018-07-20 | Ionis Pharmaceuticals Inc | MODULATE THE EXPRESSION OF APOLIPOPROTEIN (a) |
AU2016355178B9 (en) | 2015-11-19 | 2019-05-30 | Massachusetts Institute Of Technology | Lymphocyte antigen CD5-like (CD5L)-interleukin 12B (p40) heterodimers in immunity |
EP3967758A1 (en) | 2015-12-01 | 2022-03-16 | CRISPR Therapeutics AG | Materials and methods for treatment of alpha-1 antitrypsin deficiency |
WO2017096395A1 (en) | 2015-12-04 | 2017-06-08 | Ionis Pharmaceuticals, Inc. | Methods of treating breast cancer |
AU2015416656B2 (en) | 2015-12-07 | 2023-02-23 | Erasmus University Medical Center Rotterdam | Enzymatic replacement therapy and antisense therapy for Pompe disease |
US11761007B2 (en) | 2015-12-18 | 2023-09-19 | The Scripps Research Institute | Production of unnatural nucleotides using a CRISPR/Cas9 system |
AU2016376191A1 (en) | 2015-12-23 | 2018-07-12 | Crispr Therapeutics Ag | Materials and methods for treatment of amyotrophic lateral sclerosis and/or frontal temporal lobular degeneration |
AU2017205462A1 (en) | 2016-01-05 | 2018-06-07 | Ionis Pharmaceuticals, Inc. | Methods for reducing LRRK2 expression |
WO2017132483A1 (en) | 2016-01-29 | 2017-08-03 | Vanderbilt University | Free-solution response function interferometry |
AU2017213826A1 (en) | 2016-02-04 | 2018-08-23 | Curis, Inc. | Mutant smoothened and methods of using the same |
US20190112353A1 (en) | 2016-02-18 | 2019-04-18 | Crispr Therapeutics Ag | Materials and methods for treatment of severe combined immunodeficiency (scid) or omenn syndrome |
JP7033072B2 (en) | 2016-02-25 | 2022-03-09 | ザ ブリガム アンド ウィメンズ ホスピタル インコーポレイテッド | Treatment for fibrosis targeting SMOC2 |
US11136577B2 (en) | 2016-03-09 | 2021-10-05 | Ionis Pharmaceuticals, Inc. | Methods and compositions for inhibiting PMP22 expression |
WO2017158422A1 (en) | 2016-03-16 | 2017-09-21 | Crispr Therapeutics Ag | Materials and methods for treatment of hereditary haemochromatosis |
WO2017161168A1 (en) | 2016-03-16 | 2017-09-21 | Ionis Pharmaceuticals, Inc. | Modulation of dyrk1b expression |
AU2017234678A1 (en) | 2016-03-16 | 2018-08-16 | Ionis Pharmaceuticals, Inc. | Methods of modulating KEAP1 |
JP2019516393A (en) | 2016-03-18 | 2019-06-20 | カリス サイエンス インコーポレイテッド | Oligonucleotide probes and uses thereof |
ES2933435T3 (en) | 2016-04-13 | 2023-02-08 | Ionis Pharmaceuticals Inc | Methods to reduce the expression of C9ORF72 |
AU2017252023A1 (en) | 2016-04-18 | 2018-11-15 | Crispr Therapeutics Ag | Materials and methods for treatment of hemoglobinopathies |
WO2017191503A1 (en) | 2016-05-05 | 2017-11-09 | Crispr Therapeutics Ag | Materials and methods for treatment of hemoglobinopathies |
CN109414408B (en) | 2016-05-16 | 2022-03-29 | 得克萨斯州大学系统董事会 | Cationic sulfonamide amino lipids and amphiphilic zwitterionic amino lipids |
WO2017205686A1 (en) | 2016-05-25 | 2017-11-30 | Caris Science, Inc. | Oligonucleotide probes and uses thereof |
US11708614B2 (en) | 2016-06-15 | 2023-07-25 | Streck Llc | Assays and methods for determining microbial resistance |
EP3471781A4 (en) | 2016-06-17 | 2020-05-06 | Ionis Pharmaceuticals, Inc. | Modulation of gys1 expression |
WO2017223528A1 (en) | 2016-06-24 | 2017-12-28 | The Scripps Research Institute | Novel nucleoside triphosphate transporter and uses thereof |
WO2018002783A1 (en) | 2016-06-29 | 2018-01-04 | Crispr Therapeutics Ag | Materials and methods for treatment of friedreich ataxia and other related disorders |
WO2018002762A1 (en) | 2016-06-29 | 2018-01-04 | Crispr Therapeutics Ag | Materials and methods for treatment of amyotrophic lateral sclerosis (als) and other related disorders |
WO2018002812A1 (en) | 2016-06-29 | 2018-01-04 | Crispr Therapeutics Ag | Materials and methods for treatment of myotonic dystrophy type 1 (dm1) and other related disorders |
AU2017292173B2 (en) | 2016-07-06 | 2022-01-13 | Vertex Pharmaceuticals Incorporated | Materials and methods for treatment of pain related disorders |
EP3481856A1 (en) | 2016-07-06 | 2019-05-15 | Crispr Therapeutics AG | Materials and methods for treatment of pain related disorders |
WO2018007871A1 (en) | 2016-07-08 | 2018-01-11 | Crispr Therapeutics Ag | Materials and methods for treatment of transthyretin amyloidosis |
US11253601B2 (en) | 2016-07-11 | 2022-02-22 | Translate Bio Ma, Inc. | Nucleic acid conjugates and uses thereof |
RS63928B1 (en) | 2016-07-15 | 2023-02-28 | Ionis Pharmaceuticals Inc | Compounds and methods for modulation of smn2 |
WO2018020323A2 (en) | 2016-07-25 | 2018-02-01 | Crispr Therapeutics Ag | Materials and methods for treatment of fatty acid disorders |
NL2017294B1 (en) | 2016-08-05 | 2018-02-14 | Univ Erasmus Med Ct Rotterdam | Natural cryptic exon removal by pairs of antisense oligonucleotides. |
NL2017295B1 (en) | 2016-08-05 | 2018-02-14 | Univ Erasmus Med Ct Rotterdam | Antisense oligomeric compound for Pompe disease |
US11364304B2 (en) | 2016-08-25 | 2022-06-21 | Northwestern University | Crosslinked micellar spherical nucleic acids |
SG10201607303YA (en) | 2016-09-01 | 2018-04-27 | Agency Science Tech & Res | Antisense oligonucleotides to induce exon skipping |
WO2018055577A1 (en) | 2016-09-23 | 2018-03-29 | Synthena Ag | Mixed tricyclo-dna, 2'-modified rna oligonucleotide compositions and uses thereof |
JOP20190065A1 (en) | 2016-09-29 | 2019-03-28 | Ionis Pharmaceuticals Inc | Compounds and methods for reducing tau expression |
WO2018067900A1 (en) | 2016-10-06 | 2018-04-12 | Ionis Pharmaceuticals, Inc. | Method of conjugating oligomeric compounds |
SG10201609048RA (en) | 2016-10-28 | 2018-05-30 | Agency Science Tech & Res | Antisense oligonucleotides |
CA3037046A1 (en) | 2016-10-31 | 2018-05-03 | University Of Massachusetts | Targeting microrna-101-3p in cancer therapy |
JOP20190104A1 (en) | 2016-11-10 | 2019-05-07 | Ionis Pharmaceuticals Inc | Compounds and methods for reducing atxn3 expression |
US11033570B2 (en) | 2016-12-02 | 2021-06-15 | Cold Spring Harbor Laboratory | Modulation of Lnc05 expression |
EP3555296A4 (en) | 2016-12-13 | 2020-07-29 | Seattle Children's Hospital (DBA Seattle Children's Research Institute) | Methods of exogenous drug activation of chemical-induced signaling complexes expressed in engineered cells in vitro and in vivo |
IL301053A (en) | 2017-01-23 | 2023-05-01 | Regeneron Pharma | Hsd17b13 variants and uses thereof |
EP3585900B1 (en) | 2017-02-22 | 2022-12-21 | CRISPR Therapeutics AG | Materials and methods for treatment of spinocerebellar ataxia type 2 (sca2) and other spinocerebellar ataxia type 2 protein (atxn2) gene related conditions or disorders |
US11407997B2 (en) | 2017-02-22 | 2022-08-09 | Crispr Therapeutics Ag | Materials and methods for treatment of primary hyperoxaluria type 1 (PH1) and other alanine-glyoxylate aminotransferase (AGXT) gene related conditions or disorders |
WO2018154439A1 (en) | 2017-02-22 | 2018-08-30 | Crispr Therapeutics Ag | Materials and methods for treatment of spinocerebellar ataxia type 1 (sca1) and other spinocerebellar ataxia type 1 protein (atxn1) gene related conditions or disorders |
CA3054031A1 (en) | 2017-02-22 | 2018-08-30 | Crispr Therapeutics Ag | Compositions and methods for gene editing |
EP3585807A1 (en) | 2017-02-22 | 2020-01-01 | CRISPR Therapeutics AG | Materials and methods for treatment of early onset parkinson's disease (park1) and other synuclein, alpha (snca) gene related conditions or disorders |
US11180756B2 (en) | 2017-03-09 | 2021-11-23 | Ionis Pharmaceuticals | Morpholino modified oligomeric compounds |
JOP20190215A1 (en) | 2017-03-24 | 2019-09-19 | Ionis Pharmaceuticals Inc | Modulators of pcsk9 expression |
US20180284123A1 (en) | 2017-03-30 | 2018-10-04 | California Institute Of Technology | Barcoded rapid assay platform useful for efficient analysis of candidate molecules and methods of making and using the platform |
US11203611B2 (en) | 2017-04-14 | 2021-12-21 | Tollnine, Inc. | Immunomodulating polynucleotides, antibody conjugates thereof, and methods of their use |
WO2018193428A1 (en) | 2017-04-20 | 2018-10-25 | Synthena Ag | Modified oligomeric compounds comprising tricyclo-dna nucleosides and uses thereof |
JP2020517613A (en) | 2017-04-20 | 2020-06-18 | シンセナ アーゲー | Modified oligomeric compounds containing tricyclo DNA nucleosides and uses thereof |
EP3612232A1 (en) | 2017-04-21 | 2020-02-26 | The Broad Institute, Inc. | Targeted delivery to beta cells |
WO2018209270A1 (en) | 2017-05-11 | 2018-11-15 | Northwestern University | Adoptive cell therapy using spherical nucleic acids (snas) |
US11622977B2 (en) | 2017-05-12 | 2023-04-11 | Crispr Therapeutics Ag | Materials and methods for engineering cells and uses thereof in immuno-oncology |
EP3645546A4 (en) | 2017-06-30 | 2021-12-01 | Solstice Biologics, Ltd. | Chiral phosphoramidite auxiliaries and methods of their use |
CN111051512A (en) | 2017-07-11 | 2020-04-21 | 辛索克斯公司 | Incorporation of non-natural nucleotides and methods thereof |
EP3652186A4 (en) | 2017-07-13 | 2021-03-31 | Northwestern University | General and direct method for preparing oligonucleotide-functionalized metal-organic framework nanoparticles |
WO2019028425A1 (en) | 2017-08-03 | 2019-02-07 | Synthorx, Inc. | Cytokine conjugates for the treatment of autoimmune diseases |
WO2019036613A1 (en) | 2017-08-18 | 2019-02-21 | Ionis Pharmaceuticals, Inc. | Modulation of the notch signaling pathway for treatment of respiratory disorders |
US10517889B2 (en) | 2017-09-08 | 2019-12-31 | Ionis Pharmaceuticals, Inc. | Modulators of SMAD7 expression |
SG11202003464VA (en) | 2017-10-17 | 2020-05-28 | Crispr Therapeutics Ag | Compositions and methods for gene editing for hemophilia a |
US20210180091A1 (en) | 2017-10-26 | 2021-06-17 | Vertex Pharmaceuticals Incorporated | Materials and methods for treatment of hemoglobinopathies |
US20210032622A1 (en) | 2017-11-09 | 2021-02-04 | Crispr Therapeutics Ag | Self-inactivating (sin) crispr/cas or crispr/cpf1 systems and uses thereof |
TWI809004B (en) | 2017-11-09 | 2023-07-21 | 美商Ionis製藥公司 | Compounds and methods for reducing snca expression |
US20200385719A1 (en) | 2017-11-16 | 2020-12-10 | Alnylam Pharmaceuticals, Inc. | Kisspeptin 1 (kiss1) irna compositions and methods of use thereof |
EP3714054A1 (en) | 2017-11-20 | 2020-09-30 | Alnylam Pharmaceuticals, Inc. | Serum amyloid p component (apcs) irna compositions and methods of use thereof |
CN111727251A (en) | 2017-11-21 | 2020-09-29 | 克里斯珀医疗股份公司 | Materials and methods for treating autosomal dominant retinitis pigmentosa |
US20200384033A1 (en) | 2017-12-05 | 2020-12-10 | Vertex Pharmaceuticals Incorporated | Crispr-cas9 modified cd34+ human hematopoietic stem and progenitor cells and uses thereof |
WO2019118916A1 (en) | 2017-12-14 | 2019-06-20 | Ionis Pharmaceuticals, Inc. | Conjugated antisense compounds and their use |
CA3084825A1 (en) | 2017-12-14 | 2019-06-20 | Crispr Therapeutics Ag | Novel rna-programmable endonuclease systems and their use in genome editing and other applications |
JP7348185B2 (en) | 2017-12-21 | 2023-09-20 | アルニラム ファーマスーティカルズ インコーポレイテッド | Chirally enriched double-stranded RNA agent |
AU2018393050A1 (en) | 2017-12-21 | 2020-06-18 | Bayer Healthcare Llc | Materials and methods for treatment of Usher Syndrome Type 2A |
EP3728595A1 (en) | 2017-12-21 | 2020-10-28 | CRISPR Therapeutics AG | Materials and methods for treatment of usher syndrome type 2a and/or non-syndromic autosomal recessive retinitis pigmentosa (arrp) |
WO2019126641A2 (en) | 2017-12-21 | 2019-06-27 | Ionis Pharmaceuticals, Inc. | Modulation of frataxin expression |
WO2019140231A1 (en) | 2018-01-12 | 2019-07-18 | Bristol-Myers Squibb Company | Antisense oligonucleotides targeting alpha-synuclein and uses thereof |
CA3088180A1 (en) | 2018-01-12 | 2019-07-18 | Crispr Therapeutics Ag | Compositions and methods for gene editing by targeting transferrin |
MX2020007369A (en) | 2018-01-15 | 2020-10-28 | Ionis Pharmaceuticals Inc | Modulators of dnm2 expression. |
WO2019142135A1 (en) | 2018-01-19 | 2019-07-25 | Synthena Ag | Tricyclo-dna nucleoside precursors and processes for preparing the same |
US20190233816A1 (en) | 2018-01-26 | 2019-08-01 | Massachusetts Institute Of Technology | Structure-guided chemical modification of guide rna and its applications |
MA51788A (en) | 2018-02-05 | 2020-12-16 | Vertex Pharma | SUBSTANCES AND METHODS FOR TREATING HEMOGLOBINOPATHIES |
MA51787A (en) | 2018-02-05 | 2020-12-16 | Vertex Pharma | SUBSTANCES AND METHODS OF TREATMENT OF HEMOGLOBINOPATHIES |
JP7317029B2 (en) | 2018-02-12 | 2023-07-28 | アイオーニス ファーマシューティカルズ, インコーポレーテッド | Modified compounds and uses thereof |
US20210130824A1 (en) | 2018-02-16 | 2021-05-06 | Crispr Therapeutics Ag | Compositions and methods for gene editing by targeting fibrinogen-alpha |
KR20200127207A (en) | 2018-02-26 | 2020-11-10 | 신톡스, 인크. | IL-15 conjugate and uses thereof |
US11732260B2 (en) | 2018-03-02 | 2023-08-22 | Ionis Pharmaceuticals, Inc. | Compounds and methods for the modulation of amyloid-β precursor protein |
TW202000199A (en) | 2018-03-02 | 2020-01-01 | 美商Ionis製藥公司 | Modulators of IRF4 expression |
EP3768834A1 (en) | 2018-03-19 | 2021-01-27 | CRISPR Therapeutics AG | Novel rna-programmable endonuclease systems and uses thereof |
US11661601B2 (en) | 2018-03-22 | 2023-05-30 | Ionis Pharmaceuticals, Inc. | Methods for modulating FMR1 expression |
EP4051799A2 (en) | 2018-03-30 | 2022-09-07 | Rheinische Friedrich-Wilhelms-Universität Bonn | Aptamers for targeted activaton of t cell-mediated immunity |
CN116536272A (en) | 2018-04-06 | 2023-08-04 | 儿童医疗中心有限公司 | Compositions and methods for somatic reprogramming and imprinting |
SG11202008660TA (en) | 2018-04-11 | 2020-10-29 | Ionis Pharmaceuticals Inc | Modulators of ezh2 expression |
WO2019204668A1 (en) | 2018-04-18 | 2019-10-24 | Casebia Therapeutics Limited Liability Partnership | Compositions and methods for knockdown of apo(a) by gene editing for treatment of cardiovascular disease |
WO2019213571A1 (en) | 2018-05-03 | 2019-11-07 | The Trustees Of Wheaton College | Improved membranes for nanopore sensing applications |
CR20200604A (en) | 2018-05-09 | 2021-02-09 | Ionis Pharmaceuticals Inc | Compounds and methods for reducing atxn3 expression |
BR112020020957B1 (en) | 2018-05-09 | 2022-05-10 | Ionis Pharmaceuticals, Inc | Oligomeric compounds, population and pharmaceutical composition thereof and their uses |
US11833168B2 (en) | 2018-06-14 | 2023-12-05 | Ionis Pharmaceuticals, Inc. | Compounds and methods for increasing STMN2 expression |
US11332746B1 (en) | 2018-06-27 | 2022-05-17 | Ionis Pharmaceuticals, Inc. | Compounds and methods for reducing LRRK2 expression |
MX2021000922A (en) | 2018-07-25 | 2021-03-31 | Ionis Pharmaceuticals Inc | Compounds and methods for reducing atxn2 expression. |
AU2019325255A1 (en) | 2018-08-20 | 2021-04-15 | Rogcon, Inc. | Antisense oligonucleotides targeting SCN2A for the treatment of SCN1A encephalopathies |
US20210292766A1 (en) | 2018-08-29 | 2021-09-23 | University Of Massachusetts | Inhibition of Protein Kinases to Treat Friedreich Ataxia |
EP3620520A1 (en) | 2018-09-10 | 2020-03-11 | Universidad del Pais Vasco | Novel target to treat a metabolic disease in an individual |
US20220056220A1 (en) | 2018-09-14 | 2022-02-24 | Northwestern University | Programming protein polymerization with dna |
US20210332367A1 (en) | 2018-09-18 | 2021-10-28 | Alnylam Pharmaceuticals, Inc. | KETOHEXOKINASE (KHK) iRNA COMPOSITIONS AND METHODS OF USE THEREOF |
TW202023573A (en) | 2018-09-19 | 2020-07-01 | 美商Ionis製藥公司 | Modulators of pnpla3 expression |
US20210348159A1 (en) | 2018-10-17 | 2021-11-11 | Crispr Therapeutics Ag | Compositions and methods for delivering transgenes |
TW202028222A (en) | 2018-11-14 | 2020-08-01 | 美商Ionis製藥公司 | Modulators of foxp3 expression |
BR112021008967A2 (en) | 2018-11-15 | 2021-08-17 | Ionis Pharmaceuticals, Inc. | irf5 expression modulators |
TW202039841A (en) | 2018-11-21 | 2020-11-01 | 美商Ionis製藥公司 | Compounds and methods for reducing prion expression |
US20210332495A1 (en) | 2018-12-06 | 2021-10-28 | Northwestern University | Protein Crystal Engineering Through DNA Hybridization Interactions |
KR20210117271A (en) | 2018-12-21 | 2021-09-28 | 노쓰웨스턴유니버시티 | Use of annexin to prevent and treat myofascial injuryUse of annexin to prevent and treat muscle damage |
WO2020139977A1 (en) | 2018-12-26 | 2020-07-02 | Northwestern University | Use of glucocorticoid steroids in preventing and treating conditions of muscle wasting, aging and metabolic disorder |
JP2022519532A (en) | 2019-01-31 | 2022-03-24 | アイオーニス ファーマシューティカルズ, インコーポレーテッド | Modulator of YAP1 expression |
AU2020218203A1 (en) | 2019-02-06 | 2021-08-26 | Synthorx, Inc. | IL-2 conjugates and methods of use thereof |
MA54951A (en) | 2019-02-15 | 2021-12-22 | Bayer Healthcare Llc | GENE EDITING FOR HEMOPHILIA A WITH ENHANCED FACTOR VIII EXPRESSION |
AU2020227824A1 (en) | 2019-02-27 | 2021-08-26 | Ionis Pharmaceuticals, Inc. | Modulators of MALAT1 expression |
US20220175956A1 (en) | 2019-03-06 | 2022-06-09 | Northwestern University | Hairpin-like oligonucleotide-conjugated spherical nucleic acid |
CA3139919A1 (en) | 2019-03-11 | 2020-09-17 | Ochsner Health System | Microrna regulatory network as biomarkers of seizure in patients with spontaneous intracerebral hemorrhage |
SG11202109741VA (en) | 2019-03-12 | 2021-10-28 | Crispr Therapeutics Ag | Novel high fidelity rna-programmable endonuclease systems and uses thereof |
WO2020205463A1 (en) | 2019-03-29 | 2020-10-08 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulating ube3a-ats |
WO2020243644A1 (en) | 2019-05-31 | 2020-12-03 | Streck, Inc. | Detection of antibiotic resistance genes |
US11879145B2 (en) | 2019-06-14 | 2024-01-23 | The Scripps Research Institute | Reagents and methods for replication, transcription, and translation in semi-synthetic organisms |
EP3956450A4 (en) | 2019-07-26 | 2022-11-16 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulating gfap |
CN114555128A (en) | 2019-08-15 | 2022-05-27 | 新索思股份有限公司 | Combination immunooncology therapy with IL-2 conjugates |
EP4013767A4 (en) | 2019-08-15 | 2023-10-25 | Ionis Pharmaceuticals, Inc. | Linkage modified oligomeric compounds and uses thereof |
KR20220051355A (en) | 2019-08-23 | 2022-04-26 | 신톡스, 인크. | IL-15 conjugates and uses thereof |
WO2021050554A1 (en) | 2019-09-10 | 2021-03-18 | Synthorx, Inc. | Il-2 conjugates and methods of use to treat autoimmune diseases |
JP2022552249A (en) | 2019-10-14 | 2022-12-15 | アストラゼネカ・アクチエボラーグ | Modulators of PNPLA3 expression |
WO2021086623A1 (en) | 2019-10-31 | 2021-05-06 | The Trustees Of Wheaton College | Design and characterization of multilayered structures for support of lipid bilayers |
JP2022554272A (en) | 2019-11-04 | 2022-12-28 | シンソークス, インコーポレイテッド | Interleukin 10 conjugates and uses thereof |
JP2023503635A (en) | 2019-11-27 | 2023-01-31 | クリスパー・セラピューティクス・アクチェンゲゼルシャフト | Methods of synthesizing RNA molecules |
EP4077674A1 (en) | 2019-12-18 | 2022-10-26 | Alia Therapeutics S.R.L. | Compositions and methods for treating retinitis pigmentosa |
SG10201914033YA (en) | 2019-12-31 | 2021-07-29 | Wilmar International Ltd | Polypeptides with Lipase Activity and Uses Thereof |
WO2021142245A1 (en) | 2020-01-10 | 2021-07-15 | Translate Bio, Inc. | Compounds, pharmaceutical compositions and methods for modulating expression of muc5b in lung cells and tissues |
US20230057461A1 (en) | 2020-01-27 | 2023-02-23 | The U.S.A., As Represented By The Secretary, Department Of Health And Human Services | Rab13 and net1 antisense oligonucleotides to treat metastatic cancer |
TW202140787A (en) | 2020-02-28 | 2021-11-01 | 美商Ionis製藥公司 | Compounds and methods for modulating smn2 |
CA3169523A1 (en) | 2020-02-28 | 2021-09-02 | Jaume Pons | Transglutaminase-mediated conjugation |
CN116134135A (en) | 2020-04-07 | 2023-05-16 | 阿尔尼拉姆医药品有限公司 | Compositions and methods for silencing SCN9A expression |
BR112022021813A2 (en) | 2020-04-27 | 2023-01-17 | Alnylam Pharmaceuticals Inc | APOLIPOPROTEIN AND (APOE) IRNA AGENT COMPOSITIONS AND METHODS OF USE THEREOF |
EP4143321A2 (en) | 2020-05-01 | 2023-03-08 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulating atxn1 |
AR122534A1 (en) | 2020-06-03 | 2022-09-21 | Triplet Therapeutics Inc | METHODS FOR THE TREATMENT OF NUCLEOTIDE REPEAT EXPANSION DISORDERS ASSOCIATED WITH MSH3 ACTIVITY |
AU2021296622A1 (en) | 2020-06-25 | 2023-02-23 | Synthorx, Inc. | Immuno oncology combination therapy with IL-2 conjugates and anti-EGFR antibodies |
JP2023532518A (en) | 2020-06-29 | 2023-07-28 | アイオーニス ファーマシューティカルズ, インコーポレーテッド | Compounds and methods for modulating PLP1 |
TW202227102A (en) | 2020-09-22 | 2022-07-16 | 瑞典商阿斯特捷利康公司 | Method of treating fatty liver disease |
US20230392134A1 (en) | 2020-09-30 | 2023-12-07 | Crispr Therapeutics Ag | Materials and methods for treatment of amyotrophic lateral sclerosis |
EP3978608A1 (en) | 2020-10-05 | 2022-04-06 | SQY Therapeutics | Oligomeric compound for dystrophin rescue in dmd patients throughout skipping of exon-51 |
BR112023006024A2 (en) | 2020-10-09 | 2023-05-09 | Synthorx Inc | IMMUNO-ONCOLOGY THERAPIES WITH IL-2 CONJUGATES |
MX2023004029A (en) | 2020-10-09 | 2023-04-27 | Synthorx Inc | Immuno oncology combination therapy with il-2 conjugates and pembrolizumab. |
US11447521B2 (en) | 2020-11-18 | 2022-09-20 | Ionis Pharmaceuticals, Inc. | Compounds and methods for modulating angiotensinogen expression |
CA3202708A1 (en) | 2020-11-23 | 2022-05-27 | Alpha Anomeric Sas | Nucleic acid duplexes |
GB2603454A (en) | 2020-12-09 | 2022-08-10 | Ucl Business Ltd | Novel therapeutics for the treatment of neurodegenerative disorders |
WO2022174102A1 (en) | 2021-02-12 | 2022-08-18 | Synthorx, Inc. | Lung cancer combination therapy with il-2 conjugates and an anti-pd-1 antibody or antigen-binding fragment thereof |
TW202245843A (en) | 2021-02-12 | 2022-12-01 | 美商欣爍克斯公司 | Skin cancer combination therapy with il-2 conjugates and cemiplimab |
US20220288181A1 (en) | 2021-03-12 | 2022-09-15 | Northwestern University | Antiviral vaccines using spherical nucleic acids |
TW202313679A (en) | 2021-06-03 | 2023-04-01 | 美商欣爍克斯公司 | Head and neck cancer combination therapy comprising an il-2 conjugate and a pd-1 antagonist |
KR20240021218A (en) | 2021-06-11 | 2024-02-16 | 바이엘 악티엔게젤샤프트 | Novel type V RNA programmable endonuclease system |
EP4101928A1 (en) | 2021-06-11 | 2022-12-14 | Bayer AG | Type v rna programmable endonuclease systems |
CN117500815A (en) | 2021-06-18 | 2024-02-02 | Ionis制药公司 | Compounds and methods for reducing IFNAR1 expression |
WO2023285431A1 (en) | 2021-07-12 | 2023-01-19 | Alia Therapeutics Srl | Compositions and methods for allele specific treatment of retinitis pigmentosa |
WO2023034870A2 (en) | 2021-09-01 | 2023-03-09 | Ionis Pharmaceuticals, Inc. | Compounds and methods for reducing dmpk expression |
EP4144841A1 (en) | 2021-09-07 | 2023-03-08 | Bayer AG | Novel small rna programmable endonuclease systems with impoved pam specificity and uses thereof |
WO2023086292A2 (en) | 2021-11-10 | 2023-05-19 | University Of Rochester | Gata4-targeted therapeutics for treatment of cardiac hypertrophy |
WO2023086295A2 (en) | 2021-11-10 | 2023-05-19 | University Of Rochester | Antisense oligonucleotides for modifying protein expression |
GB202117758D0 (en) | 2021-12-09 | 2022-01-26 | Ucl Business Ltd | Therapeutics for the treatment of neurodegenerative disorders |
WO2023122573A1 (en) | 2021-12-20 | 2023-06-29 | Synthorx, Inc. | Head and neck cancer combination therapy comprising an il-2 conjugate and pembrolizumab |
WO2023118349A1 (en) | 2021-12-21 | 2023-06-29 | Alia Therapeutics Srl | Type ii cas proteins and applications thereof |
WO2023122750A1 (en) | 2021-12-23 | 2023-06-29 | Synthorx, Inc. | Cancer combination therapy with il-2 conjugates and cetuximab |
WO2023118068A1 (en) | 2021-12-23 | 2023-06-29 | Bayer Aktiengesellschaft | Novel small type v rna programmable endonuclease systems |
WO2023194359A1 (en) | 2022-04-04 | 2023-10-12 | Alia Therapeutics Srl | Compositions and methods for treatment of usher syndrome type 2a |
WO2023237587A1 (en) | 2022-06-10 | 2023-12-14 | Bayer Aktiengesellschaft | Novel small type v rna programmable endonuclease systems |
WO2024050261A1 (en) | 2022-08-29 | 2024-03-07 | University Of Rochester | Antisense oligonucleotide-based anti-fibrotic therapeutics |
WO2024056880A2 (en) | 2022-09-16 | 2024-03-21 | Alia Therapeutics Srl | Enqp type ii cas proteins and applications thereof |
-
1989
- 1989-12-20 US US07/453,311 patent/US5130302A/en not_active Expired - Fee Related
-
1990
- 1990-12-17 AT AT91905077T patent/ATE159027T1/en not_active IP Right Cessation
- 1990-12-17 EP EP91905077A patent/EP0506892B1/en not_active Expired - Lifetime
- 1990-12-17 WO PCT/US1990/007446 patent/WO1991009048A1/en active IP Right Grant
- 1990-12-17 AU AU73448/91A patent/AU634450B2/en not_active Ceased
- 1990-12-17 JP JP91505222A patent/JPH05507265A/en active Pending
- 1990-12-17 CA CA002071936A patent/CA2071936A1/en not_active Abandoned
- 1990-12-17 DE DE69031567T patent/DE69031567D1/en not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
ATE159027T1 (en) | 1997-10-15 |
EP0506892A4 (en) | 1993-03-31 |
EP0506892B1 (en) | 1997-10-08 |
AU7344891A (en) | 1991-07-18 |
EP0506892A1 (en) | 1992-10-07 |
AU634450B2 (en) | 1993-02-18 |
WO1991009048A1 (en) | 1991-06-27 |
JPH05507265A (en) | 1993-10-21 |
US5130302A (en) | 1992-07-14 |
DE69031567D1 (en) | 1997-11-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP0506892B1 (en) | Boronated nucleosides | |
US5659027A (en) | Boronated compounds | |
US5272250A (en) | Boronated phosphoramidate compounds | |
EP0594578B1 (en) | Oligoribonucleoside and oligodeoxyribonucleoside boranophosphates | |
US5750669A (en) | Oligonucleotide analogs with terminal 3'-3' or 5'-5' internucleotide linkages | |
CA2177952C (en) | Nucleosides and oligonucleotides containing boron clusters | |
CN103052646A (en) | Methods for the preparation of diasteromerically pure phosphoramidate prodrugs | |
JP2007504152A (en) | Novel tricyclic nucleosides or nucleotides as therapeutic agents | |
JPS59205394A (en) | 2,5-riboadenylate-morpholinoadenylate nucleotide | |
WO2001040515A1 (en) | Gapped oligomers having site specific chiral phosphorothioate internucleoside linkages | |
Shaw et al. | Oligonucleoside boranophosphate (borane phosphonate) | |
WO1996030386A1 (en) | Amidite derivatives and oligonucleotide derivatives | |
US6033909A (en) | Oligonucleotide analogs, their preparation and use | |
CA2130926C (en) | Dual action 2',5'-oligoadenylate antiviral derivatives and uses thereof | |
Bellon et al. | Sugar modified oligonucleotides: Synthesis, nuclease resistance and base pairing of oligodeoxynucleotides containing 1-(4′-thio-β-d-ribofuranosyl)-thymine | |
Baraniak et al. | Synthesis of nucleoside–amino acid conjugates containing boranephosphate, boranephosphorothioate and boranephosphoramidate linkages |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
FZDE | Discontinued |