СПОСОБЫ ЛЕЧЕНИЯ ЗЛОКАЧЕСТВЕННЫХ ОПУХОЛЕЙ С ИСПОЛЬЗОВАНИЕМ ИНГИБИТОРА ATR

Группа изобретений относится к способам выявления рака, имеющего чувствительность к соединению-ингибитору ATR в комбинации с агентом, повреждающим ДНК, и к лечению субъектов с таким выявленным раком с помощью ингибитора ATR в комбинации с агентом, повреждающим ДНК. Измеряют уровень активности CDKN1A в злокачественной опухоли пациента. Сравнивают измеренную активность CDKN1A с активностью CDKN1A в контрольной ткани или клетке. Отбирают пациента со злокачественной опухолью, идентифицированной как имеющая сниженную активность CDKN1A, для лечения с использованием ингибитора ATR IIA-7 или I-G-32 в комбинации с повреждающим ДНК средством, которое представляет собой цисплатин или гемцитабин. Изобретения позволяют осуществлять стратификацию пациентов со злокачественной опухолью для терапии с использованием ингибитора ATR в комбинации с повреждающим ДНК средством с синергическим ответом. 4 н. и 32 з.п. ф-лы, 14 ил., 5 табл., 1 пр.

СПОСОБ ОПРЕДЕЛЕНИЯ ИНГИБИТОРА, КОВАЛЕНТНО СВЯЗЫВАЮЩЕГО ЦЕЛЕВОЙ ПОЛИПЕПТИД

Изобретение относится к области биотехнологии и может быть использовано для разработки необратимого ингибитора, ковалентно связывающего целевой полипептид. Указанный способ включает: а) представление структурной модели обратимого ингибитора или модели сайта связывания в целевом полипептиде, b) идентификацию Цис-остатка в сайте связывания целевого полипептида, с) предоставление структурной модели предполагаемого необратимого ингибитора, который потенциально ковалентно связывается с целевым полипептидом, где ингибитор содержит «боеголовку», или предоставление структурной модели группы-«боеголовки», d) определение, находится ли реакционноспособная группа «боеголовки» предполагаемого ингибитора в пределах расстояния образования связи с Цис-остатком в сайте связывания целевого полипептида, или идентификацию замещаемой позиции обратимого ингибитора, к которой может быть присоединена группа-«боеголовка», способная образовывать ковалентную связь с Цис-остатком в сайте связывания целевого полипептида ...

СПОСОБ ИНДЕНТИФИКАЦИИ КЛЕТОК, ПРОЯВЛЯЮЩИХ ВОСПРИИМЧИВОСТЬ К МОДУЛЯЦИИ ПЕРЕДАЧИ СИГНАЛА, ОПОСРЕДОВАННОЙ РЕЦЕПТОРОМ ФАКТОРА РОСТА ФИБРОБЛАСТОВ ИЛИ ЕГО ВАРИАНТОМ

Изобретение относится к области биохимии. Способ идентификации клеток, которые показывают восприимчивость к ингибированию передачи сигнала, в которую вовлечены рецептор фактора роста фибробластов (FGF-R) или его вариант, включающий определение статуса фосфорилирования субстрата 2 рецептора FGF-R (FRS-2), его варианта или его фрагмента, содержащего тирозин, в биологическом образце, в качестве биомаркера такой чувствительности к ингибированию, где в отношении клеток, которые показывают фосфорилирование тирозина FRS-2, можно ожидать, что они покажут восприимчивость к ингибированию передачи сигнала с участием FGF-R. Способ идентификации фосфорилирования в FRS-2, его варианта или его фрагмента, содержащих тирозин, в качестве биомаркера для клеток, тканей или органов. Применение идентификации фосфорилирования в FRS-2, его варианта или его фрагмента, содержащего тирозин, в качестве биомаркера для клеток, тканей или органов, которые показывают гиперактивную передачу сигнала с участием FGF-R. Применение ...

ПРИМЕНЕНИЕ SGK И NEDD В КАЧЕСТВЕ МИШЕНЕЙ ДЛЯ ДИАГНОСТИЧЕСКИХ И ТЕРАПЕВТИЧЕСКИХ ЦЕЛЕЙ

... 1. Применение по меньшей мере одного вещества для обнаружения экспрессии и/или функции активированной и/или неактивной Sgk, прежде всего Sgk1 и/или Sgk3, и/или РКВ, и/или Nedd, прежде всего Nedd4-2, для диагностики заболеваний, связанных с нарушением транспорта глюкозы. 2. Применение по п.1, отличающееся тем, что субстанция представляет собой по меньшей мере одно вещества из группы антител и нуклеотидов. 3. Применение по п.2, отличающееся тем, что применяют антитела к фосфорилированным и/или нефосфорилированным последовательностям в Sgk, прежде всего Sgk1 и/или Sgk3, и/или в РКВ, и/или Nedd, прежде всего Nedd4-2. 4. Применение по п.3, отличающееся тем, что применяют антитела к по меньшей мере одной фосфорилированной и/или нефосфорилированной киназной консенсусной последовательности, прежде всего консенсусной последовательности Sgk1 в протеине Nedd, прежде всего в протеине Nedd4-2. 5. Применение по п.1, отличающееся тем, что выявляют по меньшей мере одну мутацию в Nedd, прежде всего инактивирующую ...

ПРОИЗВОДНЫЕ АЗАИНДОЛА В КАЧЕСТВЕ ИНГИБИТОРОВ ПРОТЕИНКИНАЗ

КРИСТАЛЛИЧЕСКАЯ ФОРМА (S)-N-(5-((R)-2-(2,5-ДИФТОРФЕНИЛ)-ПИРОЛИДИН-1-ИЛ)-ПИРАЗОЛО[1,5-A]ПИРИМИДИН-3-ИЛ)-3-ГИДРОКСИПИРРОЛИДИН-1-КАРБОКСАМИДА ГИДРОСУЛЬФАТА

ИДЕНТИФИКАЦИЯ, ОЦЕНКА И ЛЕЧЕНИЕ РАКОВЫХ ЗАБОЛЕВАНИЙ С ГЕНЕТИЧЕСКОЙ ИЛИ ПРИОБРЕТЕННОЙ УСТОЙЧИВОСТЬЮ К ИНГИБИТОРАМ ALK

... 1. Способ идентификации субъекта, страдающего раковым заболеванием или имеющего риск развития ракового заболевания, как имеющего повышенный риск нечувствительности к лечению ингибитором ALK, включающий:а. отбор пробы у указанного пациента; иb. анализ указанной пробы на присутствие одной или более молекул полинуклеотидов мутантных ALK, или одного или более мутантных полипептидов ALK, или того и другого,причем присутствие одной или более молекул полинуклеотидов мутантных ALK или одного, или более мутантных полипептидов ALK, или того и другого указывает на то, что указанный субъект имеет повышенный риск нечувствительности к лечению ингибитором ALK.2. Способ по п.1, в котором указанный субъект ранее не подвергался лечению ингибитором ALK, или ранее подвергался лечению ингибитором ALK, и у него развилась по меньшей мере частичная устойчивость к ингибитору ALK.3. Способ по п.1, в котором указанное раковое заболевание выбирают из группы, состоящей из анапластической крупноклеточной лимфомы, нейробластомы ...

MUTATIONEN IN DER MIT DER RESISTENZ GEGENÜBER STI-

Homogeneous fluorescent assay for kinase, phosphatase or phosphodiesterase, useful in screening for pharmaceuticals and plant-protection agents, uses polycationic polymer as quencher

Homogeneous assay for quantitative measurement of kinase, phosphatase or phosphodiesterase reactions by reacting the enzyme with a fluorescent, (de)phosphorylatable substrate (A) in presence of polycationic polymer (B) containing quencher groups. The change in phosphorylation is determined from a change in fluorescence.

TESTVERFAHREN ZUR MESSUNG DER PHOSPHORYLIERUNG

Methods

Substrate of LRRK2 and methods of assessing LRRK2 activity

A substrate for the LRRK2 protein kinase is disclosed, wherein the substrate comprises the sequence (W/F/R/K)(W/F/R/K)(R/K)(F/VV/H/R)(Y/VV/R)(S/T)(UV/I)(R/K)(R/K)(A/Y). Method for identifying a compound expected to be useful in modulating, for example inhibiting, LRRK2 protein kinase activity using said substrate are disclosed, whereby the method comprising the steps of (1) determining whether a test compound modulates, for example inhibits, the protein kinase activity of a LRRK2 polypeptide on a substrate polypeptide and (2) selecting a compound which modulates, for example inhibits, the said LRRK2 polypeptide protein kinase activity, wherein the substrate polypeptide comprises the sequence. Such a compound may be useful in treating Parkinson's Disease or Parkinsonism. The substrate polypeptide may consist or comprise the sequence WW[R/K]FYTLR[R/K]A. In a further embodiment, antibodies against LRRK2 and methods of obtaining them are disclosed.

Assay

Kinase inhibitor screening assay

Method for determining cytolysis and cytotoxicity

The invention relates to methods for detecting the cytotoxic activity of an effector, such as a cytotoxic T lymphocyte, on a non-microbial target cell. Detection of cytotoxic activity is preferably achieved by detecting adenylate kinase release using photometric methods, e.g. adenylate kinase is detected by adding ADP and detecting its conversion to ATP using a bioluminescent reagent comprising luciferin and a luciferase.

Screening Assay

Classification of kinase inhibitors using second harmonic optical techniques

Methods for measuring protein kinase and phosphatase activity

Materials and methods for analysing phosphorylation of a target polypeptide

COMPOSITIONS AND METHODS FOR ALTERING SECOND MESSENGER SIGNALING

Methods of diagnosing and treating pre-eclampsia or eclampsia.

COMPOSITIONS AND METHODS FOR ALTERING SECOND MESSENGER SIGNALING

Methods of diagnosing and treating pre-eclampsia or eclampsia.

Methods of diagnosing and treating pre-eclampsia or eclampsia.

Methods of diagnosing and treating pre-eclampsia or eclampsia.

COMPOSITIONS AND METHODS FOR ALTERING SECOND MESSENGER SIGNALING

PROCEDURE AND PREPARATIONS AROUND RESISTANCE AGAINST BIOLOGICAL OR CHEMICAL THERAPIES FOR OVERCOMING

PROCEDURE FOR THE IDENTIFICATION AND/OR VALIDATION OF REZEPTORTYROSINKINASE INHIBITORS

PROCEDURE AND DEVICE FOR MEASURING SAMPLES BY LUMINESCENCE

ENZYMATIC MEASUREMENT OF IMATINIB MESYLAT

PROCEDURE FOR THE EVALUATION OF CELLULAR PROLIFERATIVEN ILLNESSES

PROCEDURE FOR THE IDENTIFICATION OF LRRK2 INTERACTING MOLECULES AND FOR THE CLEANING OF LRRK2

PROCEDURE AND COMPOSITIONS FOR THE PROOF OF THE ACTIVATION STATUS OF MULTIPLE PROTEINS IN SINGLE CELLS

CLINICAL DIAGNOSTIC REAGENT WITH GLUCOSE-6 PHOSPHATE DEHYDROGENASE ONE (G6PDH), PROCEDURES FOR THE G6PDH-STABILISIERUNG AND USE OF A G6PDH STABILISIERERS

NEW PROCEDURE FOR CREATING PROTEIN KINASE INHIBITORS

TEST PROCEDURE FOR THE MEASUREMENT OF THE PHOSPHORYLATION

PROCEDURE FOR THE DETERMINATION OF THE REACTIVITY OPPOSITE CHK1-INHIBITOREN

PROCEDURE AND KIT FOR THE REGULATION OF THYMIDINKINASE ACTIVITY AND USE OF IT

PROCEDURE FOR THE PROGNOSIS OF THE EFFECTIVENESS OF CHEMOTHERAPY

UPS AS THE BETA CATENIN PATH MODIFYING SUBSTANCES AND OPERATING TECHNIQUE

ELISA (ENZYME LINKED IMMUNO ABSORBENT ATE) WITH HIGH THROUGHPUT FOR THE REGULATION OF RHO KINASE (ROK) - ACTIVITY

PROCEDURE FOR THE EVALUATION THE PROLIFERATIONSHEMMENDEN EFFECT OF A HEMMERS AND A PROCEDURE FOR SCREENING THE CONNECTION TO THE INHIBITION OF THE TUMORPROLIFERATION

N-Ä3-FLUORO-4 (Ä6 (METHYLOXY) - 7-Ä (3-MORPHOLIN-4 YLPROPYL) OXYÜQUINOLIN-4-YLÜOXY) PHENYLÜ N' (4FLUOROPHENYL) CYCLOPROPANE-1,1-DICARBOXAMID FOR the TREATMENT OF CANCER

PROCEDURE FOR THE CYCLIC NUCLEOTIDE REGULATION

LRRK2 POLYPEPTIDESUBSTRATE AND YOUR USES

PROCEDURE FOR THE DETERMINATION OF THE ACTIVITY OF THE CELL CYCLE REGULARIZATION FACTOR AND PROCEDURE FOR THE DIAGNOSIS OF CANCER USING THE SAME

GEFITINIB (IRESSA) TO THE TREATMENT OF CANCER

PROCEDURE FOR MEASURING GROUPS CHEMICAL TO BIOLOGICAL MOLECULES BOUND ONES,

PROCEDURE FOR THE DETERMINATION OF SECONDARY ONES MOLECULE MODIFICATIONS USING ARRAYS

THERAPEUTIC ACTIVE SUBSTANCE AGAINST NGF

PROCEDURE FOR THE RECOGNITION OF MODULATORS OF THE VEGF KINASE DOMAIN

A FAMILY OF MAP2 PROTEIN KINASES

CYTOKIN -, STRESS - AND ONCOGEN - KINASEN OF THE HUMAN PROTEIN KINASES ACTIVATED

PLKs as modifiers of the beta catenin pathway and methods of use

Methods for modulating ikkalpha activity

PAS Kinase regulates energy homeostasis

Disclosed are compositions and methods related to PAS Kinase (PASK) and various diseases and disorders associated therewith. Included are methods for treating insulin resistance, cancer, and diabetes comprising administering a composition that inhibits PAS Kinase. Further included are methods, including high throughput screening methods, for identifying test compounds that modulate PAS Kinase.

Identification of compounds suitable for treating AD

Ccr8 as modifier of branching morphogenesis and methods of use

PHOSPHORYLATED PROTEINS AND USES RELATED THERETO

Gsk3 polypeptides

CSNKS AS MODIFIERS OF THE RAC PATHWAY AND METHODS OF USE

Compositions and methods for the treatment of cancer, screening of putative anti-cancer compounds, and assessing cancer progression

AMPK PATHWAY COMPONENTS

High-throughput Enzyme-linked Immuno absorbent Assay (HT-ELISA) for identifying R Rho-kinase (ROK) modulator

Methods and compositions for the diagnosis and treatment of cancer resistant to anaplastic lymphoma kinase (ALK) kinase inhibitors

Compositions and methods for the diagnosis and treatment of a cancer that is resistant to at least one anaplastic lymphoma kinase (ALK) kinase inhibitor are provided herein. The present invention is based on the discovery of mutations within ALK that confer resistance to at least one ALK kinase inhibitor. Polynucleotides and polypeptides having at least one ALK inhibitor resistance mutation are provided and find use in methods and compositions useful in the diagnosis, prognosis, and/or treatment of diseases associated with aberrant ALK activity, more particularly, those that are resistant to at least one ALK kinase inhibitors. Methods and compositions are also provided for the identification of agents that can inhibit the kinase activity and/or reduce the expression level of the ALK resistance mutants.

Kinase substrate sensor

The present invention relates to a cytoplasmic protein complex comprising (a) a first recombinant fusion protein comprising a kinase, fused to a first interaction polypeptide and (b) a second recombinant fusion protein comprising a domain comprising a reporter phosphorylation site, whereby said domain is fused to a second interaction polypeptide. The present invention relates further to a method to detect compound-compound-interaction using said cytoplasmic protein complex, and to cells comprising such cytoplasmic protein complex.

UPs as modifiers of the beta catenin pathway and methods of use

Tumor marker and methods of use

Methods and compositions for the diagnosis and treatment of cancer resistant to anaplastic lymphoma kinase (ALK) kinase inhibitors

Compositions and methods for the diagnosis and treatment of a cancer that is resistant to at least one anaplastic lymphoma kinase (ALK) kinase inhibitor are provided herein. The present invention is based on the discovery of mutations within ALK that confer resistance to at least one ALK kinase inhibitor. Polynucleotides and polypeptides having at least one ALK inhibitor resistance mutation are provided and find use in methods and compositions useful in the diagnosis, prognosis, and/or treatment of diseases associated with aberrant ALK activity, more particularly, those that are resistant to at least one ALK kinase inhibitors. Methods and compositions are also provided for the identification of agents that can inhibit the kinase activity and/or reduce the expression level of the ALK resistance mutants.

Methods for modulating metabolic and circadian rhythms

The role of AMPK in arcadian rhythms and methods of screening for agents that modulate such rhythms are disclosed. Compositions that are useful for modulating such rhythms and uses thereof are also disclosed.

METHOD FOR CRYSTALLIZING HUMAN GSK3 AND NOVEL CRYSTAL STRUCTURE THEREOF

Multiplexed enzymatic assays

Means of examining nephropathy

Nits as modifiers of the p53 pathway and methods of use

Promls as modifiers of the p53 pathway and methods of use

Shikimate kinase from s. aureus for drug discovery

Kcnmas as modifiers of the p53 pathway and methods of use

B3galts as modifiers of the p53 pathway and methods of use

Method of identifying compounds capable of acting as agonists or antagonists of g-protein coupled receptors

Method to detect modulators of vegf kinase domain

IDENTIFICATION OF INHIBITORS OF MITOSIS

METHOD OF DETECTING AND PREVENTING ALZHEIMER'S DISEASE, PARTICULARLY AT PRODROMAL AND EARLY STAGES

Continuous-flow enzyme assay with mass spectrometry detection

Detection of cells using molecular electrophoretic change

New markers for severe progression of idiopathic scoliosis and uses thereof to stratify scoliotic patients and predict the risk of developing scoliosis

Methods of stratifying a subject having or at risk for developing adolescent idiopathic scoliosis (AIS) into diagnostically or clinically useful subclasses are provided. The stratification is based on the subject's expression and/or activity and/or PIPK1 expression and/or activity. Also provided are methods of predicting the risk of developing a scoliosis also based on the subject's expression and/or activity and/or PIPK1 expression and/or activity; and methods of increasing GiPCR signaling in cells of a subject in need thereof comprising administering to the subject's cells an effective amount of an inhibitor of PIPK1 tyrosine phosphorylation; an activator of PIPK1Y tyrosine dephosphorylation; and/or an inhibitor of PIPK1 expression and/or activity.

Compositions and methods for treating B-lymphoid malignancies

Compositions and method for inhibiting, treating, and/or preventing a B-cell neoplasm are provided. In accordance with the present invention, methods of inhibiting, treating, and/or preventing cancer in a subject are provided. In certain embodiments, the methods comprise administering to the subject a therapeutically effective amount of at least one inhibitor of the CD19-PI3K interaction. In a particular embodiment, the method further comprises the administration of at least one other anti-cancer measure, such as the administration of at least one other chemotherapeutic agent and/or administration of radiation therapy.

Treatment of cancer with TOR kinase inhibitors

TREATMENT OF CANCER WITH TOR KINASE INHIBITORS Provided herein are methods for treating or preventing ETS overexpressing castration-resistant prostate cancer, comprising administering an effective amount of a TOR kinase inhibitor to a patient having ETS overexpressing castration-resistant prostate cancer.

Methods for identifying cellular responses attributable to signaling molecule inhibition and inhibitors thereof

Gfralpha3 and its uses

Cyclic gmp dependent protein kinase as a chemotherapeutic target for antiprotozoal agents

Fluorescence-based high throughput screening assays for protein kinases and phosphatases

Diagnostics and therapeutics for glaucoma

PROTEIN KINASE PEPTIDE SUBSTRATE DETERMINATION USING PEPTIDE LIBRARIES

A method of isolating and identifying peptide substrates for a protein kinase is disclosed. The method involves a combination of size exclusion and gallium-based metal affinity chromatography. The method includes the steps of incubating a protein kinase with a peptide library in the presence of kinase reaction components, the library comprising library members; separating library members from the kinase reaction components using size exclusion chromatography to give a pool of phosphopeptides and unphosphorylated peptides; contacting the pool with immobilized gallium ions to form chelated phosphopeptides; eluting chelated phosphopeptides away from the gallium ions to give eluted phosphopeptides; sequencing the eluted phosphopeptides, whereby a preferred amino acid sequence of a preferred peptide substrate for a protein kinase is elucidated. Also disclosed is a method of identifying a compound that modulates the protein kinase catalyzed phosphorylation of a peptide substrate and a method ...

EMISSION RATIOMETRIC INDICATORS OF PHOSPHORYLATION

A chimeric phosphorylation indicator is provided. A chimeric phosphorylation indicator can contain a donor molecule, a phosphorylatable domain, a phosphoaminoacid binding domain (PAABD), and an acceptor molecule. A chimeric phosphorylation indicator also can contain a phosphorylatable polypeptide and a fluorescent protein, wherein the phosphorylatable polypeptide is contained within the sequence of the fluorescent protein, or wherein the fluorescent protein is contained within the sequence of the phosphorylatable polypeptide. Also provided are polynucleotides encoding such chimeric phosphorylation indicators, as well as kits containing the indicators or the polynucleotides. In addition, a method of using the chimeric phosphorylation indicators to detect a kinase or phosphatase in a sample is provided.

METHOD FOR MEASURING TYROSINE KINASE PHOSPHORYLATION

Method, kit and composition for measuring the autophosphorylation of one or more tyrosine kinases in presence of a kinase inhibitor compared to the absence of said kinase inhibitor for kinase specificity profiling of kinase inhibitors.

ASSAY FOR DETECTING THE ENZYMATIC ACTIVITY OF A PHOSPHORYLATION ENZYME USING ENHANCED SIGNAL GENERATION

An assay system is provided for detecting the enzymatic activity of a phosphorylation enzyme, which enzyme may be a phosphatase or protein kinase. The substrate for the enzyme is immobilized on a solid support via covalent or non-covalent binding, through a signal enhancing polymer. The immobilized substrate provides an enhanced signal to background ratio, when compared to a substrate in solution. The methods are easily adapted to high throughput screening systems.

ASSAYS FOR MEASURING PHOSPHATE MODIFICATION ENZYME ACTIVITY

The present invention relates to assays that can measure the activity of enzymes that catalyze phosphate modifications, such as kinases, phosphatases, cyclases and phosphodiesterases. The assays can also be used to identify and screen for substances that modulate the activity of kinases, phosphatases, cyclases and phosphodiesterases.

SCREENING METHODS USING C-ABL, FYN AND SYK IN COMBINATION WITH TAU PROTEIN

METHODS FOR SELECTIVELY MODULATING SURVIVIN APOPTOSIS PATHWAYS

The present invention, based on the discovery of a new biological phenomena, provides methods and compositions for use in identifying agents that modulate the phosphorylation of survivin, the interaction between survivin and p34cdc2- cyclin B1 kinase complex, and the interaction between survivin and caspase-9. Related methods and compositions can be used to modulate survivin regulated apoptosis.

HUMAN ORTHOLOGUES OF WART

The present invention relates in part to hWART nucleic acid molecules. The invention also relates in part to nucleic acid molecules encoding portions of hWART full-length proteins, nucleic acid vectors containing hWART nucleic acid molecules, recombinant cells containing such nucleic acid vectors, polypeptides purified from such recombinant cells, antibodies to such polypeptides, and methods of identifying compounds that modulate the function of an hWART polypeptide. Also disclosed are methods for diagnosing abnormal cell proliferative conditions in an organism using hWART-related molecules or compounds.

Tyrosine kinase receptor tyro3 as a therapeutic target in the treatment of cancer

The present invention concerns new methods for treating cancer by using TYRO3 inhibitors and methods for identifying new molecules of interest for treating cancer.

Irak kinase family as novel target and biomarker for alzheimer

The present invention relates to methods and devices for the diagnosis or drug response prediction of neurological disorders by measuring kinase activity and studying the phosphorylation levels and profiles in samples of said patients. Furthermore the present invention relates to methods of identifying drug compounds relevant to neurological disorders by measuring kinase activity and studying phosphorylation levels. Also, the present invention relates to the use of inhibitors of the IRAK protein kinase family or a pharmaceutical composition thereof in the treatment of neurological disorders such as Alzheimer's disease.

Methods of diagnosing cervical cancer

The invention provides reagents and methods for detecting pathogen infections in human samples. This detection utilizes specific proteins to detect the presence of pathogen proteins or abnormal expression of human proteins resulting from pathogen infections. Specific methods, compositions and kits are disclosed herein for the detection of oncogenic Human papillomavirus E6 proteins in clinical samples.

Ipp complex as marker for erlotinib treatment

The present invention provides biomarkers which are predictive for the clinical benefit of erlotinib hydrochloride treatment in cancer patients.

Inhibition of the activity of kinase and synthetase enzymes

The invention provides a method of inhibiting the activity of a kinase or a synthetase, the method including binding an active site of the kinase or synthetase with a deuterated imidazole moiety, thereby inhibiting the activity of the kinase or the synthetase.

Use of tyrosine kinase inhibitors for treatment of cushing's disease and hypercortisolism

The invention relates to methods and kits for the treatment of, prevention of, and lowering the chances of developing Cushing's Disease and/or hypercortisolism by the administration of a tyrosine kinase inhibitor, such as gefitinib.

Diagnostic marker for effect of anticancer agent

The present invention enables to realize a convenient determination of a therapeutic effect of an anticancer agent on a cancer. Specifically, the present invention provides a diagnostic marker for an effect of an anticancer agent on a cancer, comprising a substance having an affinity for a fragment of an extracellular domain of c-MET; a diagnostic reagent for an effect of an anticancer agent on a cancer, comprising a substance having an affinity for a fragment of an extracellular domain of c-MET and the fragment of the extracellular domain of c-MET; and a method of testing an effect of an anticancer agent on a cancer, comprising (a) measuring a concentration of a fragment of an extracellular domain of c-MET in a biological sample from a subject, and (b) comparing the measured concentration of the fragment of the extracellular domain of c-MET with an indicator which presents a relationship between a concentration of the fragment of the extracellular domain of c-MET and the effect of the anticancer agent on the cancer.

Post-Translational Modifications Identified by Elemental Analysis

Methods and kits for enzymes involved in post-translational modifications are provided. The methods employ elemental analysis, including ICP-MS. The methods allow for the convenient and accurate analysis of post-translation modifications of substrates by enzymes involved in post-translational modifications, including kinase and phosphatase enyzmes 1. A method for a kinase assay , the method comprising:a) incubating ATP, at least one kinase, and a free non-phosphorylated substrate labeled with an element tag, with a support having attached thereto a metal ion coordination complex under conditions to enable the kinase to phosphorylate the substrate;b) separating free non-phosphorylated substrate from phosphorylated substrate labeled with an element tag bound to the support;c) eluting the element tag associated with the resultant phosphorylated substrate into a solution; andd) performing solution elemental analysis of said solution.2. The method of wherein step (a) comprises incubating a multitude of free non-phosphorylated substrates claim 1 , each labeled with a unique element tag.3. The method of where the metal ion coordination complex attached to the support is a titanium oxide bead.4. The method of where in step (a) claim 1 , the conditions to enable the kinase to phosphorylate the substrate include a kinase reaction buffer.5. The method of wherein step (a) comprises incubating antagonists or agonists of kinase.6. The method of wherein the kinase is delivered in the form of a cell lysate.7. A method for a kinase assay claim 1 , comprising:a) incubating ATP, at least one kinase, and a free metal ion coordination complex, with an immobilized non-phosphorylated substrate under conditions which enable the kinase to phosphorylate the substrate;b) separating immobilized phosphorylated substrate attached to the metal ion coordination complex from the free ion coordination complex and the immobilized non-phosphorylated substrate;c) eluting the metal ion coordination ...

METHODS AND COMPOSITIONS FOR DETECTING PROTEIN MODIFICATIONS

Methods and compositions for detecting a protein modification in vitro and in vivo are disclosed. In certain embodiments, the protein modification detected is phosphorylation. 1. A method for the detection of a modification to a target protein , comprising:(a) contacting a target protein comprising an unnatural amino acid with a modifying enzyme; and(b) assaying for a detectable signal from the unnatural amino acid in (a) after the target protein of (a) has been contacted with the modifying enzyme,wherein if the modifying enzyme modifies the target protein, the unnatural amino acid generates a detectable signal, thereby indicating that the target protein has been modified.2. The method of claim 1 , wherein the detectable signal is fluorescence.3. The method of claim 1 , wherein the unnatural amino acid is 7HC.4. The method of claim 1 , wherein the target protein is STAT3.5. The method of claim 1 , wherein the target protein comprises an SH2 domain.6. The method of claim 1 , wherein the modifying enzyme is a kinase.7. The method of claim 1 , wherein the modification to the target protein is phosphorylation.8. The method of claim 1 , wherein the target protein comprising the unnatural amino acid is expressed in a host cell selected from the group consisting of bacterial claim 1 , insect claim 1 , and mammalian cells.9. The method of claim 8 , wherein the host cell is a bacterial cell.10E. coli. The method of claim 9 , wherein the bacterial cell is an cell.11. The method of claim 8 , wherein the host cell is a mammalian cell.12. The method of claim 11 , wherein the mammalian cell is a HepG2 cell.13. The method of claim 1 , wherein the unnatural amino acid generates a detectable signal as a result of a conformational change.14. The method of claim 1 , wherein the unnatural amino acid generates a detectable signal as a result of a pH change.15. The method of claim 8 , further comprising contacting the host cell expressing the target protein with a physiological activator ...

Methods and Tools for Screening Agents Exhibiting an Activity on Receptors of the Tumor Necrosis Factor Receptor Superfamily

The present invention provides novel chimeric receptors and methods of screening using the chimeric receptors. The chimeric receptors comprise an extracellular domain of a tumor necrosis factor receptor superfamily (TNFRSF) receptor and an intracellular domain with kinase activity stemming from a receptor tyrosine kinase. According to an embodiment, the chimeric receptor comprises a full-length TNFRSF receptor. The present invention provides means for screening and testing of modulators of TNFRSF receptors. 1. A chimeric polypeptide comprising:a first part comprising an amino acid sequence that is substantially identical to the amino acid sequence of an extracellular, ligand-binding portion of a receptor A, said receptor A being selected from receptors of the tumor necrosis factor receptor super family (TNFRSF);a second part comprising an amino acid sequence that is substantially identical to the amino acid sequence of an intracellular, signalling kinase portion of a receptor B, said receptor B being selected from receptor tyrosine kinases (RTKs); and,between said first and second parts, a third part comprising an amino acid sequence taken from and/or substantially identical to a transmembrane domain.2. The chimeric polypeptide of , wherein the amino acid sequence of said first part has the capacity of oligomerization with the corresponding extracellular domain of the receptor A and/or with another chimeric polypeptide of .3. The chimeric polypeptide of claim 1 , wherein the amino acid sequence of said first part has the capacity of binding an agent exhibiting an activity on receptor A claim 1 , such as a natural ligand of the receptor A.4. The chimeric polypeptide of claim 1 , wherein the amino acid sequence of said second part has the capacity of oligomerization with the corresponding intracellular domain of the receptor B and/or of another chimeric polypeptide of .5. The chimeric polypeptide of claim 1 , wherein the amino acid sequence of said second part has ...

Inhibition of AXL Signaling in Anti-Metastatic Therapy

Compositions and methods are provided for alleviating cancer in a mammal by administering a therapeutic dose of a pharmaceutical composition that inhibits activity of AXL protein activity, for example by competitive or non-competitive inhibition of the binding interaction between AXL and its ligand GAS6.

SCREENING METHODS

The present invention provides materials and methods relating to screening for compounds useful in the treatment of Alzheimer's disease and related conditions. In particular, screening methods using tyrosine kinases are provided, as are methods relating to the role of tyrosine kinases as therapeutic targets. 151.-. (canceled)52. An in vitro method of screening for substances which are candidate therapeutic agents for the treatment of tauopathies , said substance being effective to inhibit the phosphorylation of a tau protein by a tyrosine kinase , wherein the tau protein comprises at least one phosphorylation site , the method comprising:(a) contacting at least one said substance, the tau protein and tyrosine kinase under conditions in which the tyrosine kinase is capable of phosphorylating the site(s) of the tau protein in the absence of the substance;(b) detecting whether, and optionally the extent to which, the substance inhibits the phosphorylation of the tau protein at one or more sites of the tau protein by the tyrosine kinase; and,(c) selecting the substance which inhibits phosphorylation of the tau protein at one or more of the sites;wherein the tyrosine kinase is selected from the group consisting of Fyn or Syk.53. The method of claim 52 , wherein the tau protein is paired helical filament tau.5452. The method of claim 52 , wherein the tau protein is a fragment or derivative of a tau protein having the amino acid sequence set out in .55. The method of claim 52 , wherein the tau protein has greater than 80% sequence identity with a tau protein having the amino acid sequence set out in .56. The method of claim 52 , wherein the tyrosine kinase phosphorylates tau protein at one or more sites selected from the group consisting of Y18 claim 52 , Y29 claim 52 , Y197 claim 52 , Y310 and Y394 of tau protein.57. The method of claim 52 , wherein the tyrosine kinase is Fyn.58. The method of claim 57 , wherein Fyn phosphorylates tau protein at one or more sites selected ...

Methods for diagnosing solid tumors

An embodiment of the present invention is useful for identifying tumor cells in bladder cancer washes, pleural effusions, biliary tumor cells and circulating tumor cells in whole blood. Subsequent analysis may identify therapeutic treatments based on a single cell analysis of activatable elements in cell signaling pathways. This analysis can be useful for diagnosis, prognosis, therapy selection and monitoring of solid tumor diseases.

DISTINGUISHING CELLS IN A SAMPLE BY INACTIVATING EXTRACELLULAR ENZYME BEFORE RELEASING INTRACELLULAR ENZYME

A method for detecting the absence or presence of cells of interest in a liquid sample, wherein: (a) the sample: (i) comprises an extracellular medium containing an enzyme with a measurable activity; and (ii) is suspected of containing cells of interest that contain an enzyme with said measurable activity; and (b) the method comprises the steps of: (i) treating the liquid sample with a reagent that inactivates said measurable activity in the extracellular medium, but does not inactivate the measurable activity in said cells of interest; (ii) lysing the cells of interest to release the intracellular enzyme; and (iii) measuring said measurable activity. Thus the intracellular enzyme can be measured without interference from the extracellular enzyme. The invention is particularly useful for treatment of bacterially-infected blood using a detection assay based on adenylate kinase activity. 1. A method for detecting the absence or presence of cells of interest in a liquid sample , the method comprising: wherein the extracellular medium contains an extracellular enzyme having a measurable enzymatic activity;', 'wherein the sample is suspected of comprising cells of interest that comprise an intracellular enzyme having the measurable enzymatic activity;', 'wherein treating the liquid sample comprises treating the sample with a reagent that inactivates the measurable enzymatic activity in the extracellular medium, but does not inactivate the measurable enzymatic activity in the cells of interest;, 'treating a liquid sample comprising an extracellular medium;'}lysing said cells of interest to release the intracellular enzyme; andmeasuring the measurable enzymatic activity.2. The method of claim 1 , wherein the cells of interest are microorganism cells.3. The method of claim 2 , wherein the microorganism cells comprise bacterial cells.4. The method of claim 1 , wherein the liquid sample comprises a blood sample.5. The method of claim 4 , wherein the blood sample comprises ...

c-Src Selected Reaction Monitoring Assay

Objective quantitation of the c-Src protein directly in cancer patient tissue can aid in determining the aggressiveness of an individual patient's tumor as well as help make more informed decisions about choice of therapy. However, the c-Src protein is currently analyzed directly in formalin fixed patient tissue only by immunohistochemistry methodology which is at best subjectively semi-quantitative. This invention describes an objective quantitative assay for the c-Src protein using mass spectrometry as the analytical methodology. Specific peptides, experimentally discovered characteristics about the peptides, and experimentally established assay conditions based on those peptide characteristics are provided for use in a mass spectrometry-based Selected Reaction Monitoring (SRM) assay in order to measure relative or absolute quantitative levels of c-Src directly in a protein preparation obtained from a formalin fixed cancer patient tissue sample. 1. A method for measuring levels of the Src protein in a biological sample , comprising detecting at least one fragment peptide from Src in a protein digest prepared from said biological sample using mass spectrometry; and calculating the levels of the Src protein in said sample , wherein said measured levels of the Src protein are independently selected from a relative level or an absolute quantitative level.2. The method of claim 1 , further comprising the step of fractionating said protein digest prior to detecting said peptides.3. The method of claim 2 , wherein said fractionating step is selected from the group consisting of gel electrophoresis claim 2 , liquid chromatography claim 2 , capillary electrophoresis claim 2 , nano-reversed phase liquid chromatography claim 2 , high performance liquid chromatography claim 2 , isoelectric separation chromatography claim 2 , or reverse phase high performance liquid chromatography.4. (canceled)5. The method of claim 1 , wherein said protein digest comprises a protease digest.6 ...

CRYSTAL STRUCTURE OF HUMAN JAK3 KINASE DOMAIN COMPLEX AND BINDING POCKETS THEREOF

The present invention relates to human Janus Kinase 3 (JAK3) and JAK3-like binding pockets. The present invention provides a computer comprising a data storage medium encoded with the structure coordinates of such binding pockets. This invention also relates to methods of using the structure coordinates to solve the structure of homologous proteins or protein complexes. In addition, this invention relates to methods of using the structure coordinates to screen for and design compounds, including inhibitory compounds, that bind to JAK3 protein or JAK3 protein homologues, or complexes thereof. The invention also relates to crystallizable compositions and crystals comprising JAK3 kinase domain and JAK3 kinase domain complexes with AMP-PNP. 1. A crystal comprising a human Janus Kinase 3 kinase domain.2. A crystal comprising a Janus Kinase 3 kinase domain homologue.341-. (canceled)42. A method of using the crystal of or in an inhibitor screening assay comprising:(a) selecting a potential inhibitor by performing rational drug design with a three-dimensional structure determined for the crystal, wherein said selecting is performed in conjunction with computer modeling;(b) contacting the potential inhibitor with a kinase; and(c) detecting the ability of the potential inhibitor for inhibiting the kinase. This application claims the benefit under 35 U.S.C. §119 of U.S. Provisional Application No. 60/566,393, filed Apr. 28, 2004, and U.S. Provisional Application titled “CRYSTAL STRUCTURE OF HUMAN JAK3 KINASE DOMAIN COMPLEX AND BINDING POCKETS THEREOF” and filed Apr. 8, 2005, the disclosures of which is incorporated herein by reference.The present invention relates to human Janus Kinase 3 (JAK3) and JAK3-like binding pockets. The present invention provides a computer comprising a data storage medium encoded with the structure coordinates of such binding pockets. This invention also relates to methods of using the structure coordinates to solve the structure of homologous ...

VOLATILE ORGANIC COMPOUNDS FOR DETECTING CELL DYSPLASIA AND GENETIC ALTERATIONS ASSOCIATED WITH LUNG CANCER

The present invention provides methods of identifying a genetic abnormality such as mutation in EGFR or KRAS or ALK which is associated with the management of lung cancer or diagnosing, prognosing or monitoring the treatment of pre-cancerous conditions of the lung, such as bronchial dysplasia or atypical alveolar hyperplasia (AAH), through the detection of at least one volatile organic compound indicative of these states. 1. A method of identifying a genetic alteration selected from a mutation in EGFR , a mutation in KRAS , an ALK-ELM4 translocation and CMET amplification , wherein the genetic alteration is associated with lung cancer , the method comprising the steps of:a) obtaining a sample from a test subject;b) determining the level of at least one volatile organic compound in the test sample; andc) comparing the level of the at least one volatile organic compound from the test sample with the level of said at least one volatile organic compound in a negative control sample, whereby a significantly different level of said at least one volatile organic compound in the test sample as compared to the level of said compound in the negative control sample is indicative of the presence of said genetic alteration.2. The method according to claim 1 , wherein the at least one volatile organic compound is selected from the group consisting of 4-methyl-1-heptanol claim 1 , acetic acid octyl ester claim 1 , decane claim 1 , 3-methyl-decane claim 1 , octanal claim 1 , pentadecanenitrile claim 1 , and tetradecene; or selected from the group consisting of 4-methyl-1-heptanol claim 1 , 6-methyl-1-heptanol claim 1 , 2-ethyl-1-hexanol claim 1 , acetic acid octyl ester claim 1 , benzaldehyde claim 1 , decance claim 1 , 3-methyl-dodecance claim 1 , tetrahydrofuran claim 1 , isopropyl myristate claim 1 , octanal claim 1 , pentadecanenitrile claim 1 , 2 claim 1 ,2 claim 1 ,4-trimethyl-pentanenitrile claim 1 , 2 claim 1 ,2 claim 1 ,4-trimethyl-3-carboxyisopropyl-isobutyl ester ...

Tyrosine Kinome

Protein kinases are important signaling molecules involved in tumorigenesis. Mutational analysis of the human tyrosine kinase gene family (98 genes) identified somatic alterations in −20% of colorectal cancers, with the majority of mutations occurring in NTRK3, FES, GUCY2F and a previously uncharacterized tyrosine kinase gene called MCCK/MLK4. Most alterations were in conserved residues affecting key regions of the kinase domain. These data represent a paradigm for the unbiased analysis of signal transducing genes in cancer and provide useful targets for therapeutic intervention. 1. A method of screening test substances for use as anti-cancer agents , comprising:contacting a test substance with a protein kinase selected from the group consisting of: NTRK3, FES, MCCK, EPHA3, NTRK2, INSRR, JAK1, PDGFRA, EPHA7, EPHA8, KDR, FGFR1, and ERBB4;testing activity of the protein kinase;identifying a test substance which inhibits the activity of the protein kinase as a potential anti-cancer agent.2. The method of wherein the protein kinase is in a cell.3. The method of wherein the protein kinase is isolated from a cell.4. The method of wherein the protein kinase is in a cell of a cancer cell line.5. The method of wherein the protein kinase is in a cell which has been modified to express the protein kinase.6. The method of wherein the protein kinase is NTRK3.7. The method of wherein the protein kinase is FES.8. The method of wherein the protein kinase is MCCK.9. The method of wherein the protein kinase is EPHA3.10. The method of wherein the protein kinase is NTRK2.11. The method of wherein the protein kinase is INSRR.12. The method of wherein the protein kinase is JAK1.13. The method of wherein the protein kinase is PDGFRA.14. The method of wherein the protein kinase is EPHA7.15. The method of wherein the protein kinase is EPHA8.16. The method of wherein the protein kinase is ERBB4.17. The method of wherein the protein kinase is FGFR1.18. The method of wherein the protein kinase ...

EML4-ALK FUSION GENE

The present inventors found that a fusion gene present in some cancer patients is an oncogene. The present invention relates to a polypeptide as a novel fusion protein, a polynucleotide encoding the polypeptide, a vector comprising the polynucleotide, a transformed cell comprising the vector, a method for detecting the fusion protein or polynucleotide, a method for screening a therapeutic agent for cancer, and a method for treating cancer that is shown to be positive for the fusion gene. Further, the present invention relates kit, primer set, and probe useful in the detection of cancer that is shown to be positive for the fusion gene. 1. An isolated polypeptide comprising the amino acid sequence represented by SEQ ID NO: 2 or 7 and having a kinase activity.2. The isolated polypeptide according to comprising the amino acid sequence represented by SEQ ID NO: 2 or 7.3. An isolated polynucleotide encoding the polypeptide according to .4. An expression vector comprising the polynucleotide according to .5. A cell transformed with the expression vector according to .6. A method for producing the polypeptide according to claim 4 , comprising culturing a transformed cell according to under conditions suitable for polypeptide expression and collecting the polypeptide from the cell.7. (canceled)8. (canceled)9. A kit for detecting a fusion gene of EML4 gene and ALK gene claim 1 , comprising sense and antisense primers designed to specifically amplify a polynucleotide encoding the polypeptide according to any of .10. A primer set for detecting a fusion gene of EML4 gene and ALK gene claim 3 , comprising an antisense primer comprising nucleic acid molecule hybridizing under stringent conditions to i) the polynucleotide according to claim 3 , ii) a polynucleotide comprising the nucleotide sequence represented by SEQ ID NO: 4 claim 3 , and/or iii) a polynucleotide comprising the nucleotide sequence represented by SEQ ID NO: 5 claim 3 , and a sense primer comprising a nucleic acid ...

Assay for Identification of LRRK2 Inhibitors

A method for assessing the effect of a test compound on LRRK2 in a cell-based system, the method comprising the steps of a) assessing the effect of exposing the cell-based system comprising LRRK2 to the test compound on the phosphorylation state of Ser910 and/or Ser935 of the LRRK2; and/or b) assessing the effect of exposing the cell-based system comprising LRRK2 to the test compound on the binding of the LRRK2 to a 14-3-3 polypeptide. The method may comprise or further comprise the step of assessing the effect of exposing the cell-based system comprising LRRK2 to the test compound on the subcellular location of LRRK2. The method is considered to be useful in assessing the effect of putative LRRK2 inhibitors in cell based systems, including in vivo systems. 1. An antibody that binds specifically to LRRK2 phosphorylated at Ser910; or an antibody that binds specifically to LRRK2 phosphorylated at Ser935; or an antibody that binds specifically to LRRK2 that is not phosphorylated at Ser910; or an antibody that binds specifically to LRRK2 that is not phosphorylated at Ser935.2. A kit of parts comprising two or more of:1) an antibody that binds specifically to LRRK2 phosphorylated at Ser910, or an antibody that binds specifically to LRRK2 that is not phosphorylated at Ser910;2) an antibody that binds specifically to LRRK2 phosphorylated at Ser935, or an antibody that binds specifically to LRRK2 that is not phosphorylated at Ser935;3) a 14-3-3 polypeptide, or an antibody that specifically binds to a 14-3-3 polypeptide; and4) a fluorescently labeled LRRK2 polypeptide, or polynucleotide encoding a fluorescently labeled LRRK2.3. A purified preparation or kit of parts comprising an LRRK2 polypeptide or polynucleotide or antibody binding specifically to LRRK2; and a 14-3-3 polypeptide or polynucleotide or antibody binding specifically to a 14-3-3 polypeptide.4. A method of characterising an LRRK2 mutant , the method comprising the steps of:a) assessing the phosphorylation state ...

METHOD FOR PREDICTING DIFFERENTIATION-INDUCING PROPERTIES TO REGULATORY T-CELLS, BIOMARKER USED FOR THE METHOD, AND USE THEREOF

A method for predicting differentiation-inducing properties of naive T-cells to regulatory T-cells comprising: measuring an amount of ZAK in naive T-cells contained in the body fluid collected from the living body; and predicting differentiation-inducing properties of the naive T-cells to regulatory T-cells based on the measurement results is disclosed. A method for determining the risk of development of Graft versus Host Disease and a biomarker for predicting differentiation-inducing properties of naive T-cells to regulatory T-cells are also disclosed. 1. A method for predicting differentiation-inducing properties of naive T-cells to regulatory T-cells comprising:measuring an amount of ZAK in naive T-cells contained in the body fluid collected from the living body; andpredicting differentiation-inducing properties of the naive T-cells to regulatory T-cells based on the measurement results.2. The method according to claim 1 , wherein the differentiation-inducing properties are predicted based on the results obtained by comparing a value showing the amount of ZAK with a threshold in the prediction process.3. The method according to claim 2 , wherein the differentiation-inducing properties are predicted to be high when the value showing the amount of ZAK is higher than the threshold in the prediction process.4. The method according to claim 2 , wherein the differentiation-inducing properties are predicted to be low when the value showing the amount of ZAK is lower than the threshold in the prediction process.5. The method according to claim 1 , wherein the body fluid is cord blood claim 1 , bone marrow fluid or peripheral blood.6. The method according to claim 1 , wherein the amount of ZAK is an expression level of ZAK.7. The method according to claim 6 , wherein the expression level of ZAK is an expression level of ZAK gene or an expression level of ZAK protein.8. The method according to claim 2 , wherein the value showing the amount of ZAK is an activity value of ...

MIG6 AND THERAPEUTIC EFFICACY

We identify markers capable of guiding the decision to incorporate epidermal growth factor receptor (EGFR) inhibitors, in particular EGFR tyrosine kinase inhibitors (TKIs), into chemotherapeutic regimens. Mitogen-inducible gene 6 (Mig6), a negative regulator of EGFR, is selectively upregulated during the development of resistance to the EGFR tyrosine kinase inhibitor (TKI) erlotinib, resulting in decreased EGFR phosphorylation. The ratio of Mig6/EGFR expression highly correlates with erlotinib sensitivity. A low Mig6/EGFR ratio correlates with a high response rate to gefitinib and a marked increase in progression-free survival for patients. The ratio of Mig6 to EGFR is a major predictor of biologic and clinical responses to EGFR inhibitors. 1. A method of predicting tumor resistance to an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor , comprising:testing a patient tumor sample and determining expression level of mitogen inducible gene 6 (Mig6) and of EGFR in the sample; andcomparing the expression level of mitogen inducible gene 6 (Mig6) to the expression level of EGFR, wherein a ratio of Mig6 to EGFR lower than a predetermined cut-off value indicates sensitivity to the EGFR tyrosine kinase inhibitor and a ratio of Mig 6 higher than the predetermined cut-off value indicates resistance to the EGFR tyrosine kinase inhibitor.2. The method of wherein the predetermined cut-off value is 0.44.3. The method of wherein the expression level determined is of protein expression.4. The method of wherein the expression level determined is of mRNA expression.5. The method of wherein the tumor is selected from the group of tumors consisting of lung claim 1 , bladder claim 1 , head and neck claim 1 , and pancreatic tumors.6. The method of wherein the EGFR tyrosine kinase inhibitor is erlotinib.7. The method of wherein EGFR tyrosine kinase inhibitor is gefitinib.8. The method of wherein the inhibitor is vandetanib.9. The method of wherein the ratio is lower than ...

ASSAY SYSTEM

The invention provides a method of forming a plurality of re-constitutable doses of at least one drug in a plurality of wells, the method including the steps of (i) placing a known amount of said drug in a suitable carrier to form a first composition having a known concentration (ii) placing at least two selected amounts of that first composition into individual wells and (iii) converting the first composition into a transportable form that can later be converted into a second composition having a known concentration and (iv) sealing the wells. 1. A method of forming a plurality of re-constitutable doses of at least one drug in a plurality of wells , the method including the steps of (i) placing a known amount of said drug in a suitable carrier to form a first composition having a known concentration (ii) placing at least two selected amounts of that first composition into individual wells and (iii) converting the first composition into a transportable form that can later be converted into a second composition having a known concentration and (iv) sealing the wells.2. The method according to wherein the plurality of re-constitutable doses of at least one drug is an array of a plurality of drugs that target cell signalling molecules claim 1 , and steps (i) and (ii) include determining a series of dilutions for each of the selected drugs that span the EC50 of the molecular target of the selected drugs and dispensing an amount of each of the selected drugs into a series of wells such that when a fixed amount of the selected drugs is transferred from each well to a series of fixed volumes of the molecular target claim 1 , the final range of concentrations created spans the EC50 of the molecular target (determined previously) claim 1 , and step (iii) includes purging the series of wells with a suitable gas prior to sealing the wells.3. The method according to wherein the plurality of re-constitutable doses of at least one drug is an array of at least one drug and step (i ...

Methods and means for diagnosing spondylarthritis using autoantibody markers

The present invention relates generally to methods for diagnosing the presence or the risk of development or the therapy control of spondyloarthritis (Spa), in particular, of ankylosing spondylitis (AS) and undifferentiated spondyloarthritis in a subject, in particular in mammals. In addition, the present invention relates to test kits for use in the diagnosis of the presence or the risk of development, or for the therapy control of Spa, like AS and undifferentiated spondyloarthritis, in a subject. In particular, the present invention relates to a method for diagnosing the presence or the risk of development, or for the therapy control of Spa, like AS and undifferentiated spondyloarthritis, in a subject analysing for the presence of autoantibodies against CD74 and/or IKBKB in a subject. The presence of autoantibodies against CD74 and/or IKBKB is indicative for the presence or the risk of development, or for the therapy control of Spa, like AS and undifferentiated spondyloarthritis. In particular, detection of the presence of autoantibodies against CD74 and/or IKBKB allows early diagnosis of Spa, in particular, AS and undifferentiated spondyloarthritis.

Identification, Assessment, and Therapy of Cancers with Innate or Acquired Resistance to ALK Inhibitors

Described herein are compositions, kits, and methods for determining whether subjects having cancer(s) positive for ALK mutations are likely to respond to treatment with an ALK inhibitor and/or whether a patient having such cancer(s) is likely to have a relatively slower disease progression. Further described are methods for prognosing a time course of disease in a subject having such cancer.

Small Peptides Specifically Bind to Colorectal Cancers

Cancers are extremely heterogeneous in terms of the frequency and types of mutations present in different malignant tumors. Thus, it is likely that uniform clinical treatment is not optimal for all patients, and that the development of individualized therapeutic regimens may be beneficial. Multiple, unique small peptides bind to cell lines derived from different colon adenocarcinomas. Within two hours of contact, the colorectal cancer cells are able to transfer a P radioisotope from the small peptides to cellular proteins; the transfer occurs at a substantially higher rate than in the colorectal cancer cells than in cell lines derived from other cancers or from normal tissues. 1. A method of delivering a radioactive isotope to a colon adenocarcinoma cell , comprising:administering to a colon adenocarcinoma cell a radioactive isotope-labeled peptide, wherein the peptide is a substrate for protein kinase A (PKA) comprising the motif R-X-S/T or R-R/K-X-S/T, whereby the peptide binds to the cell and the radioactive isotope is internalized and transferred to cellular proteins.2. The method of wherein the peptide comprises a sequence R-R-X-S.3. The method of wherein the peptide comprises a sequence R-R-A/G-S.4. The method of wherein the peptide comprises a sequence S-R-R-X-S.5. The method of wherein the peptide comprises a sequence R-R-X-S-G/A.6. The method of wherein the peptide comprises a sequence G-S-R-R-X-S.7. The method of wherein the peptide comprises a sequence R-R-X-S-V.8. The method of wherein the peptide comprises a sequence R-R-X-S-V-G/A.9. The method of wherein the peptide comprises a sequence R-R-X-S selected from the group consisting of SEQ ID NO: 1-28.10. The method of wherein the peptide comprises from 4-50 amino acid residues.11. The method of wherein the peptide comprises from 4-35 amino acid residues.12. The method of wherein the peptide comprises from 4-30 amino acid residues.13. The method of wherein the peptide comprises from 9-15 amino acid ...

BIOMARKER FOR FATIGUE, AND USE THEREOF

Ratios of measured values of glucose, citrate, and cis-aconitate in a biological sample obtained from a subject to measured values of glucose, citrate, and cis-aconitate in a biological sample obtained from a healthy subject are calculated, and the glucose ratio, the citrate ratio, and the cis-aconitate ratio are used to evaluate and/or assess fatigue. Similarly, an isocitrate ratio, a succinate ratio, a malate ratio, and a lactate ratio are calculated, and these ratios are used together with the three ratios to evaluate and/or assess fatigue. This makes it possible to objectively and easily diagnose and evaluate fatigue. 1. A method for evaluating fatigue , comprising at least two steps selected from the group consisting of:(a) obtaining a first ratio of a first measured value to a first standard value, the first measured value being a measured value of a glucose concentration in a biological sample obtained from a subject;(b) obtaining a second ratio of a second measured value to a second standard value, the second measured value being a measured value of a citrate concentration in the biological sample obtained from the subject;(c) obtaining a third ratio of a third measured value to a third standard value, the third measured value being a measured value of a cis-aconitate concentration in the biological sample obtained from the subject;(d) obtaining a fourth ratio of a fourth measured value to a fourth standard value, the fourth measured value being a measured value of an isocitrate concentration in the biological sample obtained from the subject;(e) obtaining a fifth ratio of a fifth measured value to a fifth standard value, the fifth measured value being a measured value of a succinate concentration in the biological sample obtained from the subject;(f) obtaining a sixth ratio of a sixth measured value to a sixth standard value, the sixth measured value being a measured value of a malate concentration in the biological sample obtained from the subject; and(g) ...

IMIDAZOPYRIDINES SYK INHIBITORS

Certain imidazopyridines (I) and pharmaceutical compositions thereof are provided herein. Methods of treating patients suffering from certain diseases and disorders responsive to the inhibition of Syk activity, which comprises administering to such patients an amount of at least one chemical entity effective to reduce signs or symptoms of the disease or disorder are provided. Also provided are methods for determining the presence or absence of Syk kinase in a sample. 2. At least one chemical entity of claim 1 , wherein Ris chosen from phenyl claim 1 , pyridinyl claim 1 , pyrimidinyl claim 1 , pyridazinyl claim 1 , pyrazolyl claim 1 , and thiazolyl claim 1 , each of which is optionally substituted with one or more groups chosen fromhydroxy;{'sup': b', 'c', 'b', 'c, 'sub': 1', '6', '1', '4', '1', '4', '1', '4, '—NRRwherein Ris chosen from hydrogen and C-Calkyl optionally substituted with one or two groups chosen from hydroxy and —OC-Calkyl and Ris independently chosen from hydrogen and C-Calkyl optionally substituted with one or two groups chosen from hydroxy and —OC-Calkyl;'}{'sub': 3', '6', '1', '4', '1', '4', '1', '4', '1', '4', '1', '4', '2', '1', '4', '1', '4', '1', '4', '1', '4', '1', '4', '1', '4, 'heterocycloalkyl optionally substituted with one or two groups chosen from hydroxy, C-Ccycloalkyl, C-Calkyl, —C-Calkyl-OH, —C-Calkyl-O—C-Calkyl, —C-Calkyl-NH, —N(C-Calkyl)(C-Calkyl), —NH(C-Calkyl), —C(O)(C-Calkyl), —C(O)(C-Calkyl-OH), and —OC-Calkyl;'}{'sub': 1', '6', '3', '6', '1', '4', '1', '4', '1', '4', '1', '4', '1', '4', '2', '1', '4', '1', '4', '1', '4', '1', '4, '—OC-Calkyl optionally substituted with one or two groups chosen from hydroxy, C-Ccycloalkyl, C-Calkyl, —C-Calkyl-OH, —C-Calkyl-O—C-Calkyl, —C-Calkyl-NH, —N(C-Calkyl)(C-Calkyl), —NH(C-Calkyl), and —OC-Calkyl; and'}{'sub': 1', '6', '3', '6', '1', '4', '1', '4', '1', '4', '1', '4', '1', '4', '2', '1', '4', '1', '4', '1', '4', '1', '4, 'C-Calkyl optionally substituted with one or two groups chosen from ...

PFKs as Modifiers of the IGFR Pathway and Methods of Use

Human PFK genes are identified as modulators of the IGFR pathway, and thus are therapeutic targets for disorders associated with defective IGFR function. Methods for identifying modulators of IGFR, comprising screening for agents that modulate the activity of PFK are provided. 1. A method of identifying a candidate IGFR pathway modulating agent , said method comprising the steps of:(a) providing an assay system comprising a PFK polypeptide or nucleic acid;(b) contacting the assay system with a test agent under conditions whereby, but for the presence of the test agent, the system provides a reference activity; and(c) detecting a test agent-biased activity of the assay system, wherein a difference between the test agent-biased activity and the reference activity identifies the test agent as a candidate IGFR pathway modulating agent.2. The method of wherein the assay system comprises cultured cells that express the PFK polypeptide.3. The method of wherein the cultured cells additionally have defective IGFR function.4. The method of wherein the assay system includes a screening assay comprising a PFK polypeptide claim 1 , and the candidate test agent is a small molecule modulator.5. The method of wherein the assay is a kinase assay.6. The method of wherein the assay system is selected from the group consisting of an apoptosis assay system claim 1 , a cell proliferation assay system claim 1 , an angiogenesis assay system claim 1 , and a hypoxic induction assay system.7. The method of wherein the assay system includes a binding assay comprising a PFK polypeptide and the candidate test agent is an antibody.8. The method of wherein the assay system includes an expression assay comprising a PFK nucleic acid and the candidate test agent is a nucleic acid modulator.9. The method of wherein the nucleic acid modulator is an antisense oligomer.10. The method of wherein the nucleic acid modulator is a PMO.11. The method of additionally comprising:(d) administering the candidate ...

METHOD FOR PREDICTING TYROSINE KINASE INHIBITOR (TKI) RESISTANCE IN PATIENTS SUFFERING FROM CHRONIC MYELOGENOUS LEUKEMIA (CML)

The present invention relates to a method for determining or predicting the response of a patient diagnosed with chronic myelogenous leukaemia (CML) to treatment with a tyrosine kinase inhibitor. More specifically, the present invention provides methods which measure kinase activity by studying phosphorylation levels and profiles and inhibitions thereof thereby diagnosing CML patients resistant to treatment with Imatinib. 1. A method for predicting the response of a patient diagnosed with chronic myelogenous leukaemia (CML) , to a tyrosine kinase inhibitor (TKI) , comprising the steps of:(a) measuring the kinase activity of a sample, obtained from said patient diagnosed with CML, by contacting said sample with immobilized protein kinase substrates, thereby providing a phosphorylation profile of said sample, said phosphorylation profile comprising the phosphorylation levels of phosphorylation sites present in at least 10 peptide markers as listed in Table 1; and,(b) determining from said phosphorylation profile the response of said patient to said TKI.2. Method according to claim 1 , wherein said TKI is Imatinib.3. Method according to claim 1 , wherein step (b) is replaced by calculating a classifier parameter from said phosphorylation profile; and determining the expected response of said patient to treatment with said TKI on the basis of said classifier parameter.4. Method according to claim 3 , wherein said classifier parameter predicts the response of said patient to treatment with said TKI if said classifier parameter is above a first predetermined threshold level claim 3 , and wherein said classifier parameter indicates non-response of said patient to treatment with said TKI if said classifier parameter is below a second predetermined threshold level.5. Method according to claim 1 , wherein said differential phosphorylation level or said classifier parameter predicts a response claim 1 , non-response or undetermined or intermediate prediction of the effect of ...

TYROSINE KINASE BIOSENSORS AND METHODS OF USE

Disclosed are compositions and methods for measuring tyrosine kinase activity. 1. A Syk-specific biosensor comprising:{'sub': 1', '2', '3', '4', '6', '7', '8', '9, 'a peptide comprising a substrate sequence, the substrate sequence comprising a core sequence XXXXYXXXX, wherein'}{'sub': '1', 'Xis A, D, E, N, or W;'}{'sub': '2', 'Xis C, D, E, G, N, or S;'}{'sub': '3', 'Xis C, D, E, G, N, or Q;'}{'sub': '4', 'Xis D, E, F, M, S, or Y;'}{'sub': '6', 'Xis D, E, I, V, or Y;'}{'sub': '7', 'Xis D, E, H, M, N, S, or T;'}{'sub': '8', 'Xis G, L, M, or P; and'}{'sub': '9', 'Xis A, D, E, H, M, N, P, Q, S, or Y; and'}at least one of a cell penetrating peptide and an affinity tag, the cell penetrating peptide and/or tag linked directly or indirectly to the substrate sequence.2. The biosensor of claim 1 , wherein the substrate sequence is selected from the group consisting of DEEDYEEPD (SEQ ID NO:1) claim 1 , DEEDYEEPDEP (SEQ ID NO:2) claim 1 , EEDDYESPN (SEQ ID NO:3) claim 1 , EEDSYESPN (SEQ ID NO:4) claim 1 , EEDSYDSPN (SEQ ID NO:5) claim 1 , EEDDYESPNEP (SEQ ID NO:6) claim 1 , EEDSYESPNEP (SEQ ID NO:7) claim 1 , EEDSYDSPNEP (SEQ ID NO:8) claim 1 , GGEEDDYESPNEPGG (SEQ ID NO:9) claim 1 , GGEEDSYESPNEPGG (SEQ ID NO:10) claim 1 , GGEEDSYDSPNEPGG (SEQ ID NO:11) claim 1 , GGDEEDYEEPDEPGG (12) claim 1 , and GGEEDSYDSPNGG (SEQ ID NO:13).3. A kit for detecting Syk activity comprising the biosensor of and at least one of a buffer claim 1 , Syk claim 1 , and a phosphorylation detection reagent.4. The kit of claim 3 , wherein the detection reagent is selected from terbium and an antibody that preferentially binds to the peptide substrate when the tyrosine residue is phosphorylated.5. A method of measuring Syk activity in a cell comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) contacting the cell with the biosensor of under conditions suitable for phosphorylation of the biosensor by intracellular Syk; and'}(b) detecting phosphorylation of the peptide substrate.6. The method ...

Method of Modulating Protein 14-3-3 Functionality By Facilitating or Inhibiting Phosphorylation

The present invention relates to a method of modulating cellular activity. More particularly, the present invention provides a method of modulating apoptosis by modulating protein 14-3-3 phosphorylation and, thereby its functionality. The present invention still further extends to methods for identifying agents capable of modulating protein 14-3-3 phosphorylation. The method and molecules of the present invention are useful, inter alia, in the treatment and/or prophylaxis of conditions characterized by unwanted cellular activity, such as unwanted cell survival. 1. A method of modulating protein 14-3-3 or homologue or variant functionality in a mammal said method comprising administering to said mammal an effective amount of an agent for a time and under conditions sufficient to either modulate the interaction of sphingosine or homologue or variant with protein 14-3-3 or mimic the sphingosine interaction wherein agonising said interaction facilitates phosphorylation of Ser58 or analogous residue thereby inhibiting the functionality of said protein 14-3-3 and wherein antagonising said interaction inhibits phosphorylation of Ser58 or analogous residue thereby maintaining said protein 14-3-3 functionality.2. A method of modulating cellular apoptosis in a mammal said method comprising administering to said mammal an effective amount of an agent for a time and under conditions sufficient to either modulate the interaction of sphingosine or homologue or variant with protein 14-3-3 or mimic the interaction of sphingosine or homologue or variant with protein 14-3-3 wherein agonising said interaction induces apoptosis and wherein antagonising said interaction inhibits apoptosis.3. A method for the treatment or prophylaxis of a condition in a mammal , which condition is characterised by inappropriate protein 14-3-3 or homologue or variant functionality , said method comprising administering to said mammal an effective amount of an agent which either modulates the interaction ...

METHODS OF TREATING AND PREVENTING THROMBOTIC DISEASES USING ASK1 INHIBITORS

A method of treating or preventing a thrombotic disease in a subject in need thereof comprises administering to the subject an effective amount of a pharmaceutical composition comprising an inhibitor of an apoptosis signal regulating kinase 1 (ASK1) protein. A method of identifying an inhibitor of an apoptosis signal regulating kinase 1 (ASK1) protein useful for treating or preventing a thrombotic disease, comprising (a) contacting a candidate agent with a test sample comprising the ASK1 protein, and (b) comparing the ASK1 protein activity in the test sample with the ASK1 protein activity in a control sample that has not been contacted with the candidate agent, whereby a decrease in the ASK1 protein activity in the test sample compared with the control sample indicates that the candidate agent is an ASK1 inhibitor. 1. A method of treating or preventing a thrombotic disease in a subject in need thereof , comprising administering to the subject an effective amount of a pharmaceutical composition comprising an inhibitor of an apoptosis signal regulating kinase 1 (ASK1) protein.2. The method of claim 1 , wherein the thrombotic disease is selected from the group consisting of venous thrombosis claim 1 , arterial thrombosis claim 1 , atherosclerosis claim 1 , arthritis claim 1 , coagulopathy claim 1 , deep venous thrombosis (DVT) claim 1 , disseminated intravascular coagulopathy (DIC) claim 1 , pulmonary thromboembolism claim 1 , Budd-Chiari syndrome claim 1 , Paget-Schroetter diseases claim 1 , stroke and myocardial infraction.3. The method of claim 1 , wherein the ASK1 protein is obtained from activated platelets.4. The method of claim 3 , wherein the activated platelets are obtained from a subject who has suffered from the thrombotic disease.5. The method of claim 1 , wherein the ASK1 protein comprises an amino acid sequence of SEQ ID NO: 1 claim 1 , and wherein the ASK1 inhibitor is capable of attenuating phosphorylation of threonine 838 (T838) in SEQ ID NO: 1.6. The ...

EPIDERMAL GROWTH FACTOR RECEPTOR VARIANT LACKING EXON

The present invention provides an epidermal growth factor receptor variant-de4 EGFR protein. The variant lacks the fourth exon of the epidermal growth factor receptor, and promotes tumor cell invasion/metastasis. The present invention also provides an encoding gene for the variant and a method of producing the variant by means of recombination technology. 1. An isolated polypeptide of human epidermal growth factor receptor variant de4 EGFR , wherein it is selected from the following groups:(a) Polypeptide comprising the amino acid sequence of SEQ ID NO: 2;(b) Polypeptide generated by replacement, deletion or addition of amino acid sequence of SEQ ID NO: 2 with one or more amino acids, which can promote tumor cell invasion and/or migration and is derived from (a);(c) Polypeptide possessing ≧95% homology to the amino acid sequence of SEQ ID NO: 2, which can promote tumor cell invasion or migration and is derived from (a).2. The polypeptide according to of claim 1 , wherein the amino acid sequence of the polypeptide is shown as SEQ ID NO: 2.3. An isolated polynucleotide claim 1 , wherein it comprises a nucleotide sequence selected from the following groups:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) Polynucleotide encoding the polypeptide according to ;'}(b) Polynucleotide complementary to the polynucleotide (a).4. The polynucleotide according to claim 3 , wherein it encodes amino acid sequence of SEQ ID NO: 2 claim 3 , (i) Sequence of 1st-3495th in SEQ ID NO: 1;', '(ii) Sequence of 1 st-3498th in SEQ ID NO: 1., 'more preferably, the polynucleotide sequence is selected from the following groups5. A vector claim 3 , wherein it contains the polynucleotide according to .6. A genetically engineered host cell claim 3 , wherein it contains the vector according to or its chromosome is integrated by the polynucleotide according to .7. A method for preparing a polypeptide claim 3 , wherein it comprises:{'claim-ref': {'@idref': 'CLM-00006', 'claim 6'}, '(a) culturing ...

Use of the Genes in the Hog, Ras and cAMP Pathway for Treatment Of Fungal Infection

Provided herein are uses of genes for HOG, Ras and cAMP signal transduction pathways to treat fungal infection. To regulate the HOG pathway of roles of SSK1, TCO2, SSK2, PBS2, HOG1, ENA1 and NHA1 genes were investigated to find that a biosynthesis level of ergosterol is increased when these genes are inhibited. When the genes are inhibited, a large amount of ergosterol is distributed on a fungal cell membrane. Accordingly, since there are many working points of an ergosterol-binding antifungal agent, an efficiency of the ergosterol-binding antifungal agent can be considerably improved. To regulate the Ras and cAMP pathways of roles of RAS1, RAS2, CDC24, GPA1, CAC1, ACA1, PKA1, HSP12 and HSP122 genes were investigated to find that a sensitivity to a polyene- or azole-based drug is increased when these genes are inhibited. Therefore, an antifungal pharmaceutical composition including an inhibitor against the gene or protein encoded by the same can be used as an excellent combined antifungal agent which can reduce a conventional amount of an antifungal agent used and increase an efficiency. 1. A method of screening an antifungal agent for improving a fungal killing ability when used with an ergosterol-binding antifungal agent or azole-based antifungal agent , comprising:{'i': 'Cryptococcus neoformans', 'contacting Ssk1 protein of with a candidate material; and'}determining whether the candidate material inhibits or stimulates an activity of the protein.2. The method of claim 1 , wherein the ergosterol-binding antifungal agent is a polyene-based antifungal agent.3. The method of claim 2 , wherein the polyene-based antifungal agent is at least one selected from the group consisting of amphotericin B claim 2 , natamycin claim 2 , rimocidin claim 2 , filipin claim 2 , nystatin claim 2 , and candicin.4. The method of claim 2 , wherein the polyene-based antifungal agent is amphotericin B.5. The method of claim 1 , wherein the azole-based antifungal agent is at least one ...

IMIDAZOPYRAZINE SYK INHIBITORS

Certain imidazopyrazines and pharmaceutical compositions thereof are provided herein. Methods of treating patients suffering from certain diseases and disorders responsive to the inhibition of Syk activity, which comprises administering to such patients an amount of at least one chemical entity effective to reduce signs or symptoms of the disease or disorder are provided. Also provided are methods for determining the presence or absence of Syk kinase in a sample. 2. At least one chemical entity of claim 1 , wherein Ris chosen from hydrogen claim 1 , methyl claim 1 , ethyl claim 1 , and chloro.3. At least one chemical entity of claim 2 , wherein Ris hydrogen.4. At least one chemical entity of wherein Ris chosen from hydrogen and methyl.5. At least one chemical entity of claim 4 , wherein Ris hydrogen7. At least one chemical entity of claim 1 , wherein Ris phenyl substituted with one or two groups chosen fromhalo,hydroxy,carboxy,cycloalkyl optionally substituted with one or two groups chosen from hydroxy, lower alkoxy, and lower alkyl,heterocycloalkyl optionally substituted with one or two groups chosen from hydroxy, lower alkoxy, lower alkyl, lower alkyl substituted with hydroxy, optionally substituted amino, and oxo,heteroaryl,amino optionally substituted with one or two groups chosen from lower alkyl, lower alkyl substituted with halo, lower alkyl substituted with hydroxy, and lower alkyl substituted with lower alkoxy,{'sub': 6', '7', '6', '7', '6', '7, '—C(O)NRRwherein Rand Rare independently selected from hydrogen, lower alkyl, lower alkyl substituted with hydroxy, lower alkyl substituted with optionally substituted amino, cycloalkyl, aryl, heteroaryl, and heterocycloalkyl, or Rand Rtogether with the nitrogen to which they are bound form a 3- to 7-membered heterocycloalkyl ring optionally substituted with one or two groups chosen from hydroxy, lower alkyl, and lower alkyl substituted with hydroxy,'}{'sub': 2', '6', '7', '6', '7', '6', '7', '6', '7, '—S(O) ...

Methods And Compositions For The Diagnosis And Treatment Of Cancer Resistant To Anaplastic Lymphoma Kinase (ALK) Kinase Inhibitors

Compositions and methods for the diagnosis and treatment of a cancer that is resistant to at least one anaplastic lymphoma kinase (ALK) kinase inhibitor are provided herein. The present invention is based on the discovery of mutations within ALK that confer resistance to at least one ALK kinase inhibitor. Polynucleotides and polypeptides having at least one ALK inhibitor resistance mutation are provided and find use in methods and compositions useful in the diagnosis, prognosis, and/or treatment of diseases associated with aberrant ALK activity, more particularly, those that are resistant to at least one ALK kinase inhibitors. Methods and compositions are also provided for the identification of agents that can inhibit the kinase activity and/or reduce the expression level of the ALK resistance mutants.

Method for screening compounds for treating sepsis targeting nod2 signalling pathway and composition for treating sepsis comprising nod2 signalling pathway inhibitors

Methods for screening compounds for treating sepsis are disclosed. The present methods and compositions are targeting NOD2 mediated signaling pathway and the agents identified by the present methods are qualified as drug candidates for clinical development. Further Methods and composition for treating sepsis are disclosed.

Targeting mtor substrates in treating proliferative diseases

Provided are over 300 mTOR kinase targets identified by a comprehensive phosphoproteomics assay. Methods of targeting mTOR kinase targets, methods to determine the level of mTOR activity by measuring the level of phosphorylation of an mTOR targeted phosphorylation site, methods for distinguishing different classes of mTOR activity in a cell based on phosphoproteomic analysis of mTOR-targeted proteins are also provided. Also provided is the classification of a hyperproliferative disease based on phosphoproteomic analysis of mTOR-targeted proteins, as well as the personalization of therapeutic methods for the treatment of hyperproliferative disease based on phosphoproteomics. Also provided are therapeutic methods including administering to a subject an mTOR inhibitor, an mTOR inhibitor and an additional kinase inhibitor, or a dual inhibitor of mTOR and an additional kinase based on the phosphorylation levels of mTOR targets determined in the subject. Some aspects of this invention relate to the discovery that GrblO is an mTOR-targeted tumor suppressor gene.

AURORA A KINASE EFFECTORS

Two proteins (PHLDA1 and LIMK2) have been identified as direct targets of Aurora A kinase activity. PHLDA1 downregulation and Aurora A upregulation are strong predictors of poor prognosis for breast cancer patients. In accordance with one embodiment a method of detecting, prognosing and monitoring the presence/progression of cancer, and more specifically breast or prostate cancer, is provided. In one embodiment the method comprises the step of analyzing a biological sample from a patient to detect and/or quantitate the presence of Aurora A, PHLDA1 or LIMK2 amino acid sequences. In one embodiment a method of treating cancer is provided comprising the administration of therapies that enhance the activity of PHLDA1 and/or decrease the activity of LIMK2. 1. A kit for conducting Aurora A kinase reactions , said kit comprisingan Aurora A kinase;microtubule-associated protein TPX2; 'reagents for conducting a kinase reaction.', 'an Aurora A kinase substrate selected from the group consisting of PHLDA-1 and LIMK2; and'}2. The kit according to wherein the Aurora A kinase is complexed to said TPX2.3. The kit according to wherein the PHLDA-1 substrate comprises a peptide of SEQ ID NO: 1 or SEQ ID NO: 2 and the LIMK2 substrate comprises a peptide of SEQ ID NO: 3 or SEQ ID NO: 4.4. The kit according to wherein the Aurora A kinase comprises the sequence of SEQ ID NO: 5 or SEQ ID NO: 7.5. The kit according to wherein the kit further comprises labeledATP.6. The kit according to wherein the Aurora A kinase comprises the sequence of SEQ ID NO: 7.7. The kit according to wherein the kit further comprises an orthogonal ATP analog.8. The kit according to wherein the orthogonal ATP analog is N-6-Phenethyl ATP.9. The kit according to wherein the kit comprises an Aurora A kinase substrate negative control claim 4 , said negative control comprising an amino acid sequence selected from SEQ ID NO: 8 and SEQ ID NO: 9.1015.-. (canceled)16. A method of inhibiting the proliferation of cells claim 4 ...

COMPOSITIONS AND METHODS FOR TREATING CANCER AND NEURODEGENERATIVE DISEASES

The present invention relates to compositions and methods for regulating ROC-mediated death-associated protein kinase (DAPk) activity. The compositions and methods of the present invention are useful for treating or ameliorating cancer as well as pathologies associated with neuronal cell death, such as epilepsy and hypoxia/ischemia acute brain injury. The present invention further relates to screening methods for identifying agents that regulate ROC-mediated DAPk activation. 1. A method of screening for an anti cancer agent , comprising the steps of:(a) exposing a cell expressing death-associated protein kinase (DAPk) to a putative anti-cancer agent;(b) determining promotion of DAPk activation mediated by the binding of the agent to the ROC domain of DAPk;wherein promotion of DAPk activation indicates that said agent is an anti cancer agent.2. The method of claim 1 , wherein said promotion of DAPk activation is determined by the ability of the agent to negatively affect GTP binding to the ROC domain claim 1 , wherein said ability is selected from the group consisting of: prevent GTP binding to said ROC domain claim 1 , enhance GTP hydrolysis from said ROC domain claim 1 , and enhance GTP dissociation from said ROC domain of DAPk claim 1 , or wherein said promotion of DAPk activation is determined by the ability of the agent to inhibit DAPk homo-oligomerization.3. (canceled)4. A method of screening for an agent for treating pathologies associated with neuronal cell death claim 1 , comprising(a) exposing a cell expressing DAPk to a putative agent;(b) determining the inhibition or reduction DAPk activation mediated by the binding of the agent to the ROC domain of DAPk;wherein inhibition or reduction of DAPk activation indicates that said agent is capable of treating pathologies associated with neuronal cell death.5. The method of claim 4 , wherein said inhibition of DAPk activation is determined by the ability of the agent to positively affect GTP binding to the ROC ...

Activators of SGK-1 for Use as Cardioprotective Agents

Compositions for activating SGK-1 are provided. SGK-1 activators can be identified by the methods of screening provided herein. The SGK-1 activators can be used in the disclosed methods of reducing cell death and methods of treating ischemic-reperfusion injury. The ischemic-reperfusion injury can be due to a variety of complications resulting in blood loss to tissue and then the re-establishment of blood flow to the tissue.

ALK AND ROS KINASE IN CANCER

A method for identifying a patient with cancer or suspected of having cancer as a patient likely to respond to an ALK- and/or ROS-inhibiting therapeutic is provided, the method comprising: contacting a biological sample from a patient with a first reagent that specifically binds a polypeptide having ROS kinase activity and a second reagent that specifically binds to a polypeptide having ALK kinase activity, and detecting whether the first reagent or the second reagent specifically binds to the biological sample, wherein detection of binding of either the first reagent or the second reagent to the biological sample identifies the patient as a patient likely to respond to an ALK-inhibiting and/or ROS-inhibiting therapeutic. 14.-. (canceled)5. A method for identifying a patient with cancer or suspected of having cancer as a patient likely to respond to an ALK- and/or ROS-inhibiting therapeutic , comprising:contacting a biological sample from a patient with a first reagent that specifically binds a polypeptide having ROS kinase activity or specifically binds to a polynucleotide encoding a polypeptide having ROS kinase activity and a second reagent that specifically binds to a polypeptide having ALK knase activity or specifically binds to a polynucleotide encoding a polypeptide having ALK kinase activity, anddetecting whether the first reagent or the second reagent specifically binds to the biological sample, wherein detection of binding of either the first reagent or the second reagent to the biological sample identifies the patient as a patient likely to respond to an ALK- and/or ROS-inhibiting therapeutic.6. The method of claim 5 , wherein the first reagent specifically binds to full length ROS kinase protein.7. The method of claim 5 , wherein the second reagent specifically binds to full length ALK kinase protein.8. The method of claim 5 , wherein the first reagent specifically binds to the kinase domain of ROS kinase protein.9. The method of wherein the second ...

BIOLOGICAL INDICATOR

A kinase is used in a biological indicator for validation of treatment processes designed to reduce the amount or activity of a biological agent in a sample. The indication can be used for validation of sterilisation treatment. The formation of ATP from a substrate comprising ADP is measured via the liciferin/luciferate system in a luminameter. Thermostable adenylate kinase from sulfolobus acidocaldarius is especially suitable for the validation of procedures to inactivate transmissable spongiform encephalopathy agents. 183-. (canceled)84. A biological process indicator , comprising:(i) a thermostable kinase, wherein the thermostable kinase retains at least 95% of its kinase activity after exposure to 70° C. for 30 minutes; and (a) a matrix, wherein the thermostable kinase is dispersed within the matrix;', '(b) an indicator strip, a dip stick or a bead, wherein the thermostable kinase is immobilised inside the solid support or covalently coupled onto the solid support; or', '(c) a tube, wherein the thermostable kinase is attached to an internal face of the tube., '(ii) a solid support, wherein the solid support is85. The biological process indicator according to claim 84 , wherein the thermostable kinase catalyses formation of ATP from a substrate comprising ADP.86. The biological process indicator according to claim 84 , wherein the thermostable kinase retains at least 95% of its kinase activity after exposure to 80° C. for 10 minutes.87. The biological process indicator according to claim 84 , wherein the thermostable kinase is an adenylate kinase claim 84 , acetate kinase or pyruvate kinase.88. The biological process indicator according to claim 84 , wherein the thermostable kinase is a trimeric adenylate kinase or a monomeric adenylate kinase.89. The biological process indicator according to claim 84 , wherein the thermostable kinase is a trimeric adenylate kinase.90SulfolobusThermotoga. The biological process indicator according to claim 84 , wherein the ...

METHODS AND COMPOSITIONS FOR USE WITH K-RAS MEDIATED DISORDERS

Methods and compositions that can be used to identify and characterize inhibitors of K-ras localization to the plasma membrane and in doing so inhibit the signal transduction of K-ras. Such compositions can be used to treat K-ras mediated disorders, such as cancer. 1. A method of inhibiting K-ras signaling; wherein said inhibiting comprises:blocking the association of a K-ras protein with a plasma membrane, wherein said blocking is by a chemical compound.2. The method of claim 1 , wherein said compound is fendiline; fendiline HCl; or a combination thereof.3. The method of claim 2 , wherein said compound is substantially in R-isomeric form4. The method of claim 2 , wherein said blocking is by a non-L-type calcium channel blocking mechanism claim 2 , and wherein said inhibiting of K-ras signaling is Ca independent.5. The method of claim 1 , wherein said compound targets a K-ras polybasic domain.6. The method of claim 5 , wherein the polybasic domain is prenylated.7. The method of claim 1 , wherein said compound acts to perturb the electrostatic interactions of the polybasic domains and plasma membrane.8. The method of claim 1 , wherein said compound acts to perturbs the trafficking of prenylated of the polybasic domain targeted RAS proteins9. The method of claim 1 , wherein said compound reduces K-ras-plasma membrane nanoclustering; and redistributes K-ras to the endoplasmic reticulum.10. The method of claim 1 , wherein said blocking increases the binding of K-ras to at least one of: cytosol; endoplasmic reticulum; endosomes; or Golgi apparatus.11. A method of treating a K-ras mediated disorder in a patient in need thereof; said method comprising:administering to the patient a therapeutically effective amount of a compound that inhibits K-ras signaling by blocking the association of the K-ras protein with plasma membrane.12. The method of claim 11 , wherein said compound is fendiline; fendiline HCl or a combination thereof.13. The method of claim 11 , wherein said ...

Eml4-alk translocations in lung cancer

The present disclosure relates to methods for the diagnosis and evaluation of neoplastic disorders, particularly non-small cell lung cancer. Assays are described in which patient test samples are analyzed for the presence of one or more specific EML4-ALK fusion genes associated with neoplastic disorders.

NOVEL ANTIBACTERIAL COMPOUNDS

The present invention relates to compounds of formula (I): wherein Rj, R2, R3, R4, Xi, X2, X3 and Z are as defined in claim . The compounds are useful in the prevention and/or treatment of bacterial infections. 3: The compound of claim 2 , wherein Xis —O— or —NH—.5: The compound of claim 1 , wherein Xis —CH—.6: The compound of claim 1 , wherein Ris NHor OH.7: The compound of claim 1 , wherein Ris H.8: The compound of claim 1 , wherein Ris H.9: The compound of any claim 1 , wherein Ris H.10: The compound of claim 1 , wherein Ris H.11: The compound of wherein the compound is 5′-amino-5′-deoxyadenosin-8-yl-thio-N-[(5′-amino-5′-deoxyadenosine)methyl]acetamide; 8-[3-N-(5′-deoxyadenosyl)aminoprop-1-ynyl]adenosine; 8-[3-(5′-deoxyadenosyl)methoxyprop-1-ynyl]adenosine claim 1 , or any combination thereof.12: A pharmaceutical composition comprising the compound of claim 1 , in admixture with one or more pharmaceutically acceptable excipients.13: The compound of claim 1 , wherein the compound is suitable for prevention claim 1 , treatment claim 1 , or both prevention and treatment of a bacterial infection.16: A chemical probe comprising the compound of claim 1 , coupled to a detectable label.17. (canceled)18: A method for screening a molecule inhibiting NAD kinase claim 1 , the method comprising:{'claim-ref': {'@idref': 'CLM-00016', 'claim 16'}, 'contacting NAD kinase with the chemical probe of with a molecule to be screened;'}determining a quantity of chemical probe bound or unbounded to the NAD kinase; anddeducing from the quantity if the molecule is an inhibitor of the NAD kinase.19: A method of treating a bacterial infection claim 1 , the method comprising administering the compound of to a subject in need thereof. The present invention relates to compounds for use in the prevention and/or treatment of bacterial infections, pharmaceutical compositions comprising them, and processes for the preparation thereof.The search for new antibacterial compounds has become ...

METHOD FOR SUPPRESSING RECEPTOR TYROSINE KINASE-MEDIATED PRO-SURVIVAL SIGNALING IN CANCER CELL

In order to provide a screening method for a compound capable of suppressing a mechanism of ROR1 by targeting ROR1 and to provide a drug comprising as an active ingredient a compound capable of suppressing the mechanism, it has been found out that suppressing a function of ROR1 makes it possible to suppress receptor tyrosine kinase-mediated pro-survival signaling in a cancer cell. Further, it has been found out that suppressing a function of ROR1 also effectively suppresses growth of cancer cell lines having acquired resistance to inhibitors of receptor tyrosine kinases such as EGFR and MET. 1. A method for suppressing receptor tyrosine kinase-mediated pro-survival signaling in a cancer cell , the method comprising suppressing a function of ROR1.2. The method according to claim 1 , wherein the receptor tyrosine kinase is any one of EGFR and MET.3. The method according to claim 1 , wherein the cancer cell is a cancer cell resistant to any one of an EGFR tyrosine kinase inhibitor and a MET tyrosine kinase inhibitor.4. The method according to claim 1 , wherein the function of ROR1 is suppressed by suppressing a binding between ROR1 and c-Src claim 1 , suppressing phosphorylation of c-Src due to ROR1 claim 1 , suppressing a binding between ROR1 and EGFR claim 1 , suppressing a binding between EGFR and ErbB3 due to ROR1 claim 1 , suppressing phosphorylation of ErbB3 due to ROR1 claim 1 , suppressing autophosphorylation of ROR1 claim 1 , or suppressing an expression of an ROR1 gene.5. A screening method for a compound capable of suppressing receptor tyrosine kinase-mediated pro-survival signaling in a cancer cell claim 1 , the method comprising:(a) a step of bringing a test compound into contact with a system capable of detecting a function of ROR1; and(b) a step of selecting a compound having an activity of suppressing a function of ROR1.6. The method according to claim 5 , wherein the receptor tyrosine kinase is any one of EGFR and MET.7. The method according to claim 5 ...

MULTIPLE MECHANISMS FOR MODULATION OF THE P13 KINASE PATHWAY

An embodiment of the present invention is a method for measuring the post translational states and expression levels of proteins in the PI3K and/or mTOR for use in diagnosis, prognosis and drug screening applications. 1. A method for classifying a cell comprising:contacting the cell with a PI3K and/or mTOR pathway modulator;determining the presence or absence of a change in activation level of an activatable element in the cell; andclassifying the cell based on the presence or absence of the change in the activation level of the activatable element.2. The method of claim 1 , wherein the change in activation level of the activatable element is an increase in activation level of the activatable element.3. The method of claim 1 , wherein the cell is a cancer cell or hematopoietic cell.4. The method of claim 1 , wherein the presence or absence of a change in the activation level of the activatable element is compared to a normal cell contacted with the PI3K and/or mTOR inhibitor.5. The method of claim 1 , wherein the presence or absence of a change in the activation levels of the activatable element is determined in the determining step.6. The method of claim 1 , wherein the classification comprises classifying the cell as a cell that is correlated with a clinical outcome.7. The method of claim 6 , wherein the clinical outcome is the presence or absence of a cancer claim 6 , metabolic disorder or immune disorder.8. The method of claim 6 , wherein the clinical outcome is the staging or grading of a neoplastic condition.9. The method of claim 1 , wherein the classification further comprises determining a method of treatment.10. The method of claim 1 , wherein the modulator is a cancer cell modulator or hematopoietic cell modulator.11. The method of claim 1 , wherein the modulator is a growth factor claim 1 , chemokine claim 1 , cytokine claim 1 , drug claim 1 , immune modulator claim 1 , ion claim 1 , neurotransmitter claim 1 , adhesion molecule claim 1 , hormone claim 1 ...

Rapid Bioluminescence Detection System

An assay is provided for detecting the activity of a reporter kinase comprising (i) adding said reporter kinase to an assay mixture wherein said reporter kinase is contacted with bioluminescent reagent no more than 5 minutes after being contacted with ADP, and wherein, prior to contacting the reporter kinase with ADP, the assay mixture is substantially free from kinase other than reporter kinase; and (ii) detecting light output from the assay mixture. Methods for detecting the presence of an analyte in a sample and methods for validating a treatment process using the above assay are also provided. Further provided are devices for conducting these assays and methods. 126-. (canceled)27. A method of validating a treatment process for reducing the amount or activity of a contaminating biological agent in a sample , comprising the steps of:(i) providing a sample that contains, or is suspected to contain, a contaminating biological agent;(ii) subjecting the sample to a treatment process in the presence of a defined amount of an exogenous kinase, wherein the exogenous kinase and the contaminating biological agent are both exposed to the treatment process;(iii) measuring the residual activity of the exogenous kinase comprising adding the exogenous kinase to an assay mixture, wherein said exogenous kinase is contacted simultaneously with ADP and a bioluminescent reagent, wherein prior to contacting the exogenous kinase with ADP, the assay mixture is substantially free from kinase other than exogenous kinase; and(iv) detecting light output from the assay mixture, thereby measuring the residual activity of the exogenous kinase; and(v) comparing said residual activity to a predetermined kinase activity, wherein the pre-determined kinase activity corresponds to a confirmed reduction in the amount or activity of the contaminating biological agent under the same conditions.28. The method according to claim 27 , wherein prior to contacting the exogenous kinase with the ADP claim ...

KINASE SUBSTRATE SENSOR

The disclosure relates to a cytoplasmic protein complex comprising: (a) a first recombinant fusion protein comprising a kinase, fused to a first interaction polypeptide; and (b) a second recombinant fusion protein comprising a domain comprising a reporter phosphorylation site, whereby the domain is fused to a second interaction polypeptide. The disclosure relates further to a method to detect compound-compound-interaction using the cytoplasmic protein complex, and to cells comprising such cytoplasmic protein complex. 1. A cytoplasmic protein complex comprising:a first recombinant fusion protein comprising a kinase fused to a first interaction polypeptide; anda second recombinant fusion protein comprising a domain comprising a reporter phosphorylation site, wherein the domain is fused to a second interaction polypeptide.2. The cytoplasmic protein complex according to claim 1 , wherein the reporter phosphorylation site is phosphorylated.3. A cytoplasmic protein complex comprising:a first recombinant fusion protein comprising a mutant kinase fused to a first interaction polypeptide; anda second recombinant fusion protein comprising a domain comprising a reporter phosphorylation site, wherein the domain is fused to a second interaction polypeptide.4. The cytoplasmic protein complex according to claim 3 , wherein the mutant kinase is a mutant constitutive tyrosine kinase.5. The cytoplasmic protein complex according to claim 3 , wherein the mutant kinase is an inactive tyrosine kinase that is activated by addition of an exogenous small compound.6. The cytoplasmic protein complex according to claim 4 , wherein the tyrosine kinase is selected from the group consisting of a Tyk2 mutant claim 4 , a JAK mutant claim 4 , and a Src mutant.7. The cytoplasmic protein complex according to claim 1 , wherein the first recombinant fusion protein comprises SEQ ID NO:1 or SEQ ID NO:2.8. The cytoplasmic protein complex according to claim 1 , wherein the reporter phosphorylation domain is ...

NOVEL PEPTIDIC ACTIVATORS OF TYPE I cGMP DEPENDENT PROTEIN KINASES AND USES THEREOF

The present invention relates to cGMP protein kinase (PKG) and regulatory domains and methods of use thereof. The structural determination of PKG domains is also described. cGMP independent PKG activators and uses thereof are also described. 1. (canceled)2. A method of treating a PKG deficient condition in a subject , comprising administering to the subject a therapeutically effective amount of a cGMP independent PKG activator.3. The method of claim 2 , wherein the PKG deficient condition is selected from the group consisting of cardiovascular disorders claim 2 , hypoxia claim 2 , spinal cord injury claim 2 , and stroke.4. The method of claim 2 , further comprising administering cGMP to the subject.5. The method of claim 2 , further comprising administering a cGMP dependent PKG activator to the subject.6. The method of claim 5 , wherein the cGMP dependent PKG activator is a PDE inhibitor.7. The method of claim 6 , wherein the PDE inhibitor is selected from the group consisting of sildenafil citrate claim 6 , vinpocetine claim 6 , EHNA claim 6 , anagrelide claim 6 , enoximone claim 6 , milrinone claim 6 , mesembrine claim 6 , rolipram claim 6 , ibudilast claim 6 , piclamilast claim 6 , luteolin claim 6 , drotaverine claim 6 , roflumilast claim 6 , tadalafil claim 6 , vardenafil claim 6 , udenafil claim 6 , avanafil claim 6 , and papaverine.8. The method of claim 5 , wherein the a cGMP dependent PKG activator is a NO-donor.9. The method of claim 8 , wherein the NO-donor is selected from the group consisting of isosorbide dinitrite claim 8 , Diazeniumdiolates claim 8 , NONOates claim 8 , S-Nitrosothiols claim 8 , NO hybrid drugs claim 8 , and Zeolites.10. The method of claim 2 , wherein the cGMP independent PKG activator is a peptide.12. The method of claim 2 , wherein the cGMP independent PKG activator is a small molecule.13. A composition comprising: a cGMP independent PKG activator and a carrier.14. The composition of claim 13 , wherein the cGMP independent PKG ...

ALZHEIMER'S DISEASE-SPECIFIC ALTERATIONS OF PROTEIN KINASE C EPSILON (PKC-EPSILON) PROTEIN LEVELS

The present invention relates to methods of diagnosing Alzheimer's Disease in a human patient by detecting alterations in the ratio of PKC epsilon protein levels in a human patient compared with PKC epsilon levels in a control subject. The Alzheimer's disease-specific molecular biomarkers disclosed herein are useful for the diagnosis of Alzheimer's disease and for screening methods for the identification of compounds for treating or preventing Alzheimer's disease. The present invention also provides methods for elevating PKC epsilon protein levels comprising the steps of contacting one or more human cells with an amount of a PKC activator effective to elevate PKC epsilon levels compared to an uncontacted human cell. 1) A method of diagnosing Alzheimer's Disease in a human subject , said method comprising the steps of:a) determining the PKC epsilon level in said human subject; andb) comparing the PKC epsilon level in said human subject to the PKC epsilon level in a control subject;wherein said method is indicative of Alzheimer's Disease in said human subject if the PKC epsilon level in said human subject is lower than the PKC epsilon level in said control subject.2) The method of claim 1 , wherein said PKC epsilon level is measured in one or more cells.3) The method of claim 1 , wherein said PKC epsilon level is a PKC epsilon protein level or a PKC epsilon activity level.4) The method of claim 1 , wherein said PKC epsilon level is measured by RT-PCR.5) The method of claim 1 , wherein said control subject does not have Alzheimer's Disease.6) The method of claim 1 , wherein said determining step (a) is done in vitro.7) The method of claim 2 , wherein said cell is a fibroblast claim 2 , buccal mucosal claim 2 , neuron claim 2 , or blood cell.8) The method of wherein said determining step (a) comprises a method selected from the group consisting of radioimmunoassay claim 1 , Western blot assay claim 1 , immunofluorescent assay claim 1 , enzyme immunoassay claim 1 , ...

Ack1 kinase inhibition to treat triple negative breast cancer

Compositions and methods are disclosed for treating Triple Negative Breast Cancers (TNBCs). The methods involve administering to a subject with TNBC a composition containing an Ack1 kinase inhibitor. In some embodiments, the method involves first assaying a sample from the subject for Tyr176-phosphorylated-AKT and/or Tyr284-phosphorylated-Ack1. In these embodiments, detection of the phosphorylated AKT and/or Ack1 is an indication that the subject is a suitable candidate for treatment with the Ack1 kinase inhibitor.

METHODS FOR PROTEIN PURIFICATION AND ANALYSIS

Methods to separate complex protein mixtures into individual proteins while maintaining protein function or enzyme activity are disclosed. They are designed to provide high resolution and efficient recovery of the functional proteins. The purified proteins may be analyzed with functional assays including enzymatic activities. 1. A method for purifying and characterizing proteins from a mixture comprising:passing the mixture through at least two orthogonal separations under conditions that preserve protein activity;eluting the purified proteins into individual wells of a protein elution plate; andassaying the purified proteins in each well for protein activity.2. The method according to claim 1 , wherein the proteins in the mixture are purified in a first separation according to their isoelectric points.3. The method according to claim 2 , wherein the first separation utilizes no reducing agents.4. The method according to claim 1 , wherein the proteins in the mixture are purified in a second separation according to their molecular weight.5. The method according to claim 4 , wherein the second separation utilizes no more than about 2% SDS.6. The method according to claim 4 , wherein the second separation utilizes no more than about 1% SDS.7. The method according to claim 4 , wherein the second separation utilizes no more than about 0.1% SDS.8. The method according to claim 1 , wherein the purified proteins are assayed for NAD reductase activity.9. The method according to claim 1 , wherein the purified proteins are assayed for protein kinase activity.10. The method according to claim 1 , wherein the protein mixture is obtained from healthy cells.11. The method according to claim 1 , wherein the protein mixture is obtained from diseased cells.12. The method according to claim 1 , further comprising quantifying the purified proteins.13. The method according to claim 1 , further comprising identifying the purified proteins.14. The method according to claim 11 , wherein ...

JAK/STAT INHIBITORS AND MAPK/ERK INHIBITORS FOR RSV INFECTION

The present invention concerns a method for treating or reducing the likelihood of developing a respiratory syncytial virus (RSV) infection in a subject by administering an effective amount of an inhibitor of the janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway or the mitogen-activated kinase (MAPK)/extracellular signal-regulated kinase (ERK1/2) signaling pathway to the subject. Another aspect of the invention concerns a pharmaceutical composition that includes an inhibitor of JAK/STAT or MAPK/ERK signaling to the subject; and a pharmaceutically acceptable carrier. Another aspect of the invention concerns a method for identifying agents useful for treating or reducing the likelihood of developing an RSV infection 1. A method for identifying agents useful for treating or reducing the likelihood of developing a respiratory syncytial virus (RSV) infection , comprising determining whether a candidate agent acts as an inhibitor of janus kinase (JAK)/signal transducers and activators of transcription (STAT) signaling pathway or the mitogen-activated kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling pathway , wherein inhibition of JAK/STAT or MAPK/ERK is indicative of an agent useful for treating or reducing the likelihood of developing RSV infection.2. The method of claim 1 , wherein the candidate agent is a polynucleotide.3. The method of claim 1 , wherein the candidate agent is a polypeptide.4. The method of claim 1 , wherein the candidate agent is an antibody.5. The method of claim 1 , wherein the candidate agent is a small molecule.6. The method of claim 1 , wherein the candidate agent is a polynucleotide claim 1 , and wherein the polynucleotide encodes a polypeptide which claim 1 , when expressed claim 1 , inhibits JAK/STAT or MAPK/ERK signaling.7. The method of claim 1 , wherein the candidate agent is a polynucleotide claim 1 , and wherein the polynucleotide is an antisense molecule or a small interfering ...

Photoluminescent molecular complex and method for determining of the concentration of said molecular complex

The invention concerns novel molecular complexes with photoluminescent probes whose specific association with purine-binding proteins leads to increased emission of long lifetime luminescence, and the application of the probes for monitoring activity of protein kinases (PKs) and other purine-binding proteins, screening of compounds as inhibitors of PKs and characterization of inhibitors targeted to the kinase, and methods of manufacturing of such probes. The invention concerns also the use of the improved method for monitoring activity of protein kinases in living cells, characterization of inhibitors of protein kinases, analysis of protein kinase-based disease biomarkers and other tasks of biological and medical importance.

METHOD OF SCREENING FOR CHAPERONIN MODULATOR

The present invention relates to a method of screening for modulator of chaperonin that is involved in protein aggregation inducing neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and Huntington's disease, use of the chaperonin modulator screened by the method for prevention and treatment of neurodegenerative diseases. According to the present invention, novel negative chaperonin modulator is provided, and chaperonin modulator may be more rapidly and conveniently screened with the negative modulator as a target. Furthermore, by using the screened material, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and Huntington's disease may be effectively prevented or treated without concern for cell death due to autophagy, which is the existing method of removing protein aggregate. 1. A method of screening for a chaperonin modulator comprising1) preparing a candidate material and VRK2;2) measuring an expression amount or a kinase activity of the VRK2; and3) treating the VRK2 with the candidate material and measuring a change in the expression amount or the kinase activity of the VRK2, and4) determining the candidate material as the chaperonin modulator by comparing the expression amount or the kinase activity of VRK2 between the treated and untreated groups.2. The method according to claim 1 , wherein the chaperonin modulator increases an expression amount or a stability of chaperonin protein claim 1 , andthe step 4) is performed by identifying the candidate material as a material that increases the expression amount or the stability of chaperonin protein if the expression amount or the kinase activity of VRK2 is decreased compared to untreated group.3. The method according to claim 1 , wherein the chaperonin modulator decreases an expression amount or a stability of chaperonin protein claim 1 , andthe step 4) is performed by identifying the candidate material as a material that decreases the expression amount or ...

METHOD FOR THE REGULATION OF PROTEIN KINASE ACTIVITY IN VIVO AND IN VITRO

Disclosed herein is a substantially pure nucleic acid encoding a eukaryotic protein kinase having at least one mutated amino-acid residue located in its subdomain IX. Also disclosed is a substantially pure eukaryotic protein kinase polypeptide having at least one mutation in its subdomain IX, the kinase being encoded by the nucleic acid. 1. A temperature-sensitive eukaryotic protein kinase polypeptide comprising at least one mutated amino acid residue located in its subdomain IX.2. The polypeptide according to claim 1 , wherein the at least one mutated residue is the conserved aspartate and/or the residue immediately C-terminal to the conserved glycine residue in the subdomain IX.3. (canceled)4. The polypeptide according to claim 2 , wherein the conserved aspartate residue is mutated to an asparaqine or a glutamine residue.5. The polypeptide according to claim 2 , wherein the residue immediately C-terminal to the conserved glycine residue is mutated to an aspartic acid or a qlutamic acid residue.6. (canceled)7. The polypeptide according to claim 1 , wherein the subdomain IX comprises one or the following amino acid sequences SEQ ID NO: 1; SEQ ID NO:2; SEQ ID NO: 3; SEQ ID NO: 4; SEQ ID NO: 5; SEQ ID NO: 6; SEQ ID NO: 7; SEQ ID NO: 8; SEQ ID NO: 9; SEQ ID NO: 10; SEQ ID NO: 11; or SEQ ID NO: 12.89-. (canceled)10. The polypeptide according to claim 1 , wherein the kinase is of the following family of kinases: AMPK claim 1 , Aurora kinases claim 1 , BUB1 claim 1 , CAK1 claim 1 , Cyclin-dependent kinases (CDKs) Calmodulin kinase claim 1 , Casein kinases 1 and 2 claim 1 , CHK1 claim 1 , CHK2 claim 1 , MAP kinases claim 1 , MAP kinase kinase claim 1 , MPS1 claim 1 , NDR claim 1 , or p21-activated kinase.11. A method of producing a mutated eukaryotic protein kinase claim 1 , the method comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'a) transfecting a cell with a nucleic acid encoding the polypeptide according to ; and'}b) culturing the transfected cells under ...

METHODS FOR DETECTING ENZYME ACTIVITY USING FLUORESCENCE LIFETIME IMAGING

Disclosed are methods of detecting enzymatic activity on a fluorophore-labeled substrate using by monitoring the fluorescence lifetime of the fluorophore. 1. A method of detecting phosphorylation activity of an enzyme in a sample , the method comprising:contacting the sample with a biosensor, the biosensor comprising a fluorophore and a substrate of the enzyme, the substrate of the enzyme selected from a synthetic polypeptide, a peptidomimetic, and a nucleic acid mimetic; andmonitoring fluorescence lifetime of the fluorophore to detect phosphorylation of the substrate.2. The method of claim 1 , wherein the fluorescence lifetime of the fluorophore is monitored by fluorescence lifetime spectroscopy and/or fluorescence lifetime imaging microscopy (FLIM).3. The method of claim 2 , wherein the substrate of the enzyme is a synthetic polypeptide.4. The method of claim 2 , wherein the substrate of the enzyme is a peptidomimetic.5. The method of claim 1 , wherein the sample comprises an intact cell.6. The method of claim 5 , wherein the biosensor further comprises a cell penetrating peptide.7. The method of claim 6 , wherein the cell penetrating peptide is a TAT peptide.8. The method of claim 1 , wherein the enzyme is a serine claim 1 , threonine claim 1 , or tyrosine kinase.9. The method of claim 8 , wherein the enzyme is a tyrosine kinase.10. The method of claim 9 , wherein the enzyme is an Abl kinase.11. The method of claim 1 , wherein the fluorophore is an organic moiety.12. The method of claim 1 , wherein the fluorophore is selected from Cy series of dyes claim 1 , AlexaFluor series of dyes claim 1 , DyLight series of dyes claim 1 , fluorescein claim 1 , fluorescein derivatives claim 1 , rhodamine claim 1 , rhodamine derivatives claim 1 , coumarin claim 1 , coumarin derivatives claim 1 , nitrobenzoxadiazole claim 1 , nitrobenzoxadiazole derivatives claim 1 , BODIPY claim 1 , BODIPY derivatives claim 1 , pyrene claim 1 , pyrene derivatives claim 1 , dansyl claim 1 , ...

PHOSPHATASE COUPLED KINASE ASSAY

Assays and methods for detecting and quantifying the activity of a test kinase. The assay includes a nucleotide phosphatase at least ten times more active on ADP than on ATP and a free phosphate detector. The assay may also include a control kinase and/or ATP. Methods of the invention include combining a test kinase, a substrate of the test kinase, and a known quantity of ATP under conditions to produce ADP, combining the produced ADP with a phosphatase specific for ADP, and measuring free phosphate. The amount of phosphate produced directly correlates to the activity of the kinase.phosphatase coupled kinase assay 1. An assay for detecting activity of a test kinase comprising:a nucleotide phosphatase, wherein the phosphatase is at least ten times more active on ADP than on ATP;a free phosphate detector; anda control kinase.2. The assay of wherein the phosphatase is an ectonucleoside triphosphate diphosphohydrolase.3. The assay of wherein the phosphatase is ectonucleoside triphosphate diphosphohydrolase 6.4. The assay of wherein the phosphatase is at least about 30 times more active on ADP than on ATP.5. The assay of wherein the phosphatase is at least about 70 times more active on ADP than on ATP.6. The assay of further comprising a source of free phosphate for use as a phosphate standard.7. The assay of wherein the free phosphate detector comprises a colorimetric assay.8. The assay of wherein the free phosphate detector comprises a first reagent and a second reagent claim 7 , wherein the first reagent comprises ammonium molybdate and the second reagent comprises malachite green oxalate.9Saccharomyces cerevisiae. The assay of wherein the control kinase comprises a hexokinase.10. The assay of further comprising ATP.11. An assay for detecting kinase activity comprising:ATP;a nucleotide phosphatase, wherein the phosphatase is at least ten times more active on ADP than on ATP; anda free phosphate detector.12. The assay of wherein the phosphatase is an ectonucleoside ...

Benzoxazepines as Inhibitors of PI3K/mTOR and Methods of Their Use and Manufacture

The invention is directed to Compounds of Formula I: (I) and pharmaceutically acceptable salts or solvates thereof, as well as methods of treating using the compounds, methods for screening for inhibitor compounds and methods for identifying treatment regimens. 1. A method for treating a subject having a tumor comprising:(a) administering a PI3K-α selective inhibitor, a dual PI3K-α/mTOR selective inhibitor, or a combination of a PI3K-α selective inhibitor and a mTOR selective inhibitor to the subject if said tumor comprises a mutation in a PI3K-α kinase domain; or(b) administering a combination of a PI3K-α selective inhibitor and a PI3K-β selective inhibitor, a dual PI3K-α/mTOR selective inhibitor, or a PI3K-β selective inhibitor, to said subject if said tumor comprises a mutation in a PI3K-α helical domain.281-. (canceled)82. The method of claim 1 , further comprising administering an additional chemotherapeutic agent in steps (a) or (b) claim 1 , wherein said additional chemotherapeutic agent comprises anti-microtubule agents; platinum coordination complexes; alkylating agents; antibiotic agents; topoisomerase II inhibitors; antimetabolites; topoisomerase I inhibitors such as camptothecins; hormones and hormonal analogues; signal transduction pathway inhibitors; non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; proapoptotic agents; and cell cycle signaling inhibitors.83. The method of claim 82 , wherein said additional therapeutic agent administered in step (b) further comprises a PI3K-δ selective inhibitor claim 82 , a PI3K-γ selective inhibitor or a pan PI3K selective inhibitor claim 82 , wherein said pan PI3K selective inhibitor comprises PI-103 or PIK-75.84. The method according to claim 1 , wherein said mutation in said PI3K-α comprises a mutation in a kinase domain; wherein the mutation in said kinase domain comprises a mutation at position 1047 of SEQ ID NO: 1; and wherein said mutation of said kinase domain is a substitution ...

USE OF PHOSPHO-AKT AS A BIOMARKER OF DRUG RESPONSE

Use of phospho-Akt as a biomarker for predicting the response, such as resistance, to a compound, wherein phospho-Akt is Akt that has been phosphorylated on one or more residues, with the proviso that for Akt1, Akt2, and Akt3 the designation phospho-Akt is used to indicate phosphorylation at a site other than T308, T309 or T305 respectively, wherein the compound is a compound of general formula (I) wherein R represents phenyl, thienyl or pyridinyl wherein phenyl is optionally substituted by one or two substituents independently selected from alkyl, halo-lower alkyl, hydroxy-lower alkyl, lower alkoxy-lower alkyl, acyloxy-lower alkyl, phenyl, hydroxy, lower alkoxy, hydroxy-lower alkoxy, lower alkoxy-lower alkoxy, phenyl-lower alkoxy, lower alkylcarbonyloxy, amino, monoalkylamino, dialkylamino, lower alkoxycarbonylamino, lower alkylcarbonylamino, substituted amino wherein the two substituents on nitrogen form together with the nitrogen heterocyclyl, lower alkylcarbonyl, carboxy, lower alkoxycarbonyl, cyano, halogen, and nitro; and wherein two adjacent substituents are methylenedioxy; and wherein pyridinyl is optionally substituted by lower alkoxy, amino or halogen; X represents a group C═Y, wherein Y stands for oxygen or nitrogen substituted by hydroxy or lower alkoxy; Rrepresents hydrogen, lower alkylcarbonyl, hydroxy-lower alkyl or cyano-lower alkyl; R, Rand Rrepresent hydrogen; Rand R, independently of each other, represent hydrogen, lower alkyi or lower alkoxy; or Rand Rtogether represent methylenedioxy; and pharmaceutically acceptable derivatives thereof; or wherein R represents phenyl or pyridinyl wherein phenyl is optionally substituted by one or two substituents independently selected from alkyi, halo-lower alkyi, hydroxy-lower alkyi, lower alkoxy-lower alkyi, acyloxy-lower alkyi, phenyl, hydroxy, lower alkoxy, hydroxy-lower alkoxy, lower alkoxy-lower alkoxy, phenyl-lower alkoxy, lower alkylcarbonyloxy, amino, monoalkylamino, dialkylamino, lower ...

MODULATION OF PHOSPHATIDYLINOSITOL-5-PHOSPHATE-4-KINASE ACTIVITY

The invention features methods for identifying compounds that modulate the activity of phosphatidylinositol 5-phosphate 4-kinase (PI5P4K) Inhibitors of PI5P4K can be used in, for example, the treatment or prevention of cell proliferation dis orders (e.g., the prevention of tumor cell growth in p53 mutated cancers). 1. A method for identifying compounds that modulate phosphatidylinositol-5-phosphate-4-kinase (PI5P4K) , said method comprising:(a) providing a medium comprising said PI5P4K and a substrate;(b) contacting said medium with a candidate compound;(c) detecting the activity of said PI5P4K; and(d) determining if said candidate compound modulates said PI5P4K.2. The method of claim 1 , wherein said substrate is guanosine-5′-triphosphate (GTP).3. The method of claim 1 , wherein said PI5P4K is the PI5P4Kγ isoform.4. The method of claim 1 , wherein said substrate is adenosine-5′-triphosphate (ATP).5. The method of claim 1 , wherein said PI5P4K is the PI5P4Kα isoform or the PI5P4Kβ isoform.6. The method of claim 1 , wherein said method further comprises comparing the activity detected in step (c) with the detected activity of PI5P4K in a medium not contacted with said candidate compound of step (b) claim 1 , wherein(i) increased activity of PI5P4K in the presence of said candidate compound of step (b) identifies said candidate compound as a promoter of PI5P4K activity; and(ii) wherein decreased activity of PI5P4K in the presence of said candidate compound of step (b) identifies said candidate compound as an inhibitor of PI5P4K activity.7. The method of claim 1 , wherein said method further comprises comparing the activity detected in step (c) with the activity observed in a medium where said PI5P4K is absent.8. The method of claim 1 , wherein said medium of step (a) is a medium suitable for use in qualitative high throughput screening.9. The method of claim 1 , wherein said PI5P4K is human recombinant PI5P4K.10. The method of claim 1 , wherein said medium of step (a) ...

NEW BORANOPHOSPHATE ANALOGUES OF CYCLIC NUCLEOTIDES

The present invention relates to novel boranophosphate analogues of cyclic nucleotides. The invention further relates to the use of such compounds as reagents for signal transduction research or as modulators of cyclic nucleotide-regulated binding proteins and isoenzymes thereof, and/or as hydrolysis- and oxidation-resistant ligands for affinity chromatography, for antibody production or for diagnostic applications e.g. on chip surfaces and/or as additive for organ transplantation storage solutions. 3. The compound of claim 1 , whereinR1 is H, F, Cl, Br, I, azido, nitro, 2-furyl, 3-furyl, 2-bromo-5-furyl, 2-thienyl, 3-thienyl, allyl, trifluoromethyl, phenyl, OH, methoxy, ethoxy, n-propoxy, n-butoxy, benzyloxy, 4-methylbenzyloxy, 4-methoxybenzyloxy, 4-fluorobenzyloxy, 4-chlorobenzyloxy, 4-bromobenzyloxy, phenyloxy, SH, methylthio, ethylthio, n-propylthio, n-butylthio, n-pentylthio, n-hexylthio, isopropylthio, 2-hydroxyethylthio, 2-aminoethylthio, 2-carboxyethylthio, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)ethylthio, (4-bromo-2,3-dioxobutyl)thio, [2-[(fluoresceinylthioureido)amino]ethyl]thio, cyclohexylthio, benzylthio, 4-azidobenzylthio, phenylthio, 2-chlorophenylthio, 3-chlorophenylthio, 4-chlorophenylthio, 2-bromophenylthio, 3-bromophenylthio, 4-bromophenylthio, 2-fluorophenylthio, 3-fluorophenylthio, 4-fluorophenylthio, 4-nitrophenylthio, 2-aminophenylthio, 3-aminophenylthio, 4-aminophenylthio, phenylethylamino, 3-phenyl-propylamino, 2-methoxyphenylthio, 3-methoxyphenylthio, 4-methoxyphenylthio, 2-methylphenylthio, 3-methylphenylthio, 4-methylphenylthio, 2-isopropylphenylthio, 4-isopropylphenylthio, 2,3,5,6-tetrafluorophenylthio, 4-hydroxyphenylthio, 2,3-dichlorophenylthio, 2,4-dichlorophenylthio, 2,5-dichlorophenylthio, 2,4-difluorophenylthio, 2,5-dimethoxyphenylthio, 2,5-dimethylthiophenylthio, 2,6-dimethylthiophenylthio, 2,6-dichlorophenylthio, 4-isopropyloxyphenylthio, 4-methylthiophenylthio, 4-azidophenylthio, 4-methylcumarinyl, naphtyl-2-thio, 4- ...

ENZYME AND RECEPTOR MODULATION

Covalent conjugates of an α,α-disubstituted glycine ester and a modulator of the activity of a target intracellular enzyme or receptor, wherein the ester group of the conjugate is hydrolysable by one or more intracellular carboxylesterase enzymes to the corresponding acid and the α,α-disubstituted glycine ester is conjugated to the modulator at a position remote from the binding interface between the inhibitor and the target enzyme or receptor pass into cells and the active acid hydrolysis product accumulates within the cells. 1. A covalent conjugate of an α ,α-disubstituted glycine ester and a modulator of the activity of a target intracellular enzyme or receptor , wherein:the ester group of the conjugate is hydrolysable by one or more intracellular carboxylesterase enzymes to the corresponding acid; andthe α,α-disubstituted glycine ester is conjugated to the modulator at a position remote from the binding interface between the inhibitor and the target enzyme or receptor.2. A covalent conjugate of an α ,α-disubstituted glycine ester and a modulator of the activity of a target intracellular enzyme or receptor , wherein:the ester group of the conjugate is hydrolysable by one or more intracellular carboxylesterase enzymes to the corresponding acid; andthe α,α-disubstituted glycine ester is conjugated to the modulator such that the binding mode of the conjugated modulator and the said corresponding acid to the target enzyme or receptor is the same as that of the unconjugated modulator.3. A method of increasing or prolonging the intracellular potency and/or residence time of modulator of the activity of a target intracellular enzyme or receptor comprising structural modification of the modulator by covalent attachment thereto of an α ,α-disubstituted glycine ester at a position remote from the binding interface between the inhibitor and the target enzyme or receptor , the ester group of the conjugate being hydrolysable by one or more intracellular carboxylesterase ...

CRYSTALLINE FORM OF (S)-N-(5-((R)-2-(2,5-DIFLUOROPHENYL)-PYRROLIDIN-1-YL)-PYRAZOLO[1,5-A]PYRIMIDIN-3-YL)-3-HYDROXYPYRROLIDINE-1-CARBOXAMIDE HYDROGEN SULFATE

A novel crystalline form of (S)—N-(5-((R)-2-(2,5-difluorophenyl)pyrrolidin-1-yl)-pyrazolo[1,5-a]pyrimidin-3-yl)-3-hydroxypyrrolidine-1-carboxamide, pharmaceutical compositions containing said crystalline form and the use of said crystalline form in the treatment of pain, cancer, inflammation, neurodegenerative disease or infection are disclosed. In some embodiments, the novel crystalline form comprises a stable polymorph of (S)—N-(5-((R)-2-(2,5-difluorophenyl)pyrrolidin-1-yl)-pyrazolo[1,5-a]pyrimidin-3-yl)-3-hydroxypyrrolidine-1-carboxamide hydrogen sulfate. The present invention is further directed to a process for the preparation of the novel crystalline form. 1. (canceled)3. The method of claim 2 , wherein the crystalline form is characterized by having an X-ray powder diffraction pattern comprising peaks at °2θ values of 10.7±0.2 claim 2 , 18.4±0.2 claim 2 , 20.7±0.2 claim 2 , 23.1±0.2 claim 2 , and 24.0±0.2.4. The method of claim 2 , wherein the crystalline form is characterized by having an X-ray powder diffraction pattern comprising peaks at °2θ values of 10.7±0.2 claim 2 , 18.4±0.2 claim 2 , 19.2±0.2 claim 2 , 20.2±0.2 claim 2 , 20.7±0.2 claim 2 , 21.5±0.2 claim 2 , 23.1±0.2 claim 2 , and 24.0±0.2.5. The method of claim 2 , wherein the crystalline form is characterized by having an X-ray powder diffraction pattern comprising peaks at °2θ values of 10.7±0.2 claim 2 , 15.3±0.2 claim 2 , 16.5±0.2 claim 2 , 18.4±0.2 claim 2 , 19.2±0.2 claim 2 , 19.9±0.2 claim 2 , 20.2±0.2 claim 2 , 20.7±0.2 claim 2 , 21.5±0.2 claim 2 , 22.1±0.2 claim 2 , 23.1±0.2 claim 2 , 24.0±0.2 claim 2 , 24.4±0.2 claim 2 , 25.6±0.2 claim 2 , 26.5±0.2 claim 2 , 27.6±0.2 claim 2 , 28.2±0.2 claim 2 , 28.7±0.2 claim 2 , 30.8±0.2 claim 2 , and 38.5±0.2.6. The method of claim 2 , wherein the cancer is selected from the group consisting of: non-small cell lung cancer claim 2 , papillary thyroid carcinoma claim 2 , glioblastoma multiforme claim 2 , acute myeloid leukemia claim 2 , colorectal ...

COMPOSITIONS AND METHODS INVOLVING ENGINEERED P27

The disclosure provides polypeptides comprising an engineered p27, or a fragment thereof. Such polypeptides may be used to form trimeric protein complexes with a cyclin-dependent kinase 4 (Cdk4) (or a variant thereof) or Cdk6 (or a variant thereof), and a cyclin D (CycD) or a variant thereof. 1. A polypeptide comprising an engineered p27 , or a fragment thereof , wherein the engineered p27 has at least one amino acid substitution at a position selected from the group consisting of Y74 , Y88 , and Y89 , wherein the engineered p27 forms a trimeric protein complex with (i) a cyclin-dependent kinase 4 (Cdk4) or a variant thereof , or a Cdk6 or a variant thereof , and (ii) a cyclin D (CycD) or a variant thereof , and wherein the amino acid positions are determined with reference to the sequence of SEQ ID NO: 1.2. The polypeptide of claim 1 , wherein the amino acid substitution at position Y74 is Y74E claim 1 , Y74D claim 1 , or Y74R claim 1 , the amino acid substitution at Y88 is Y88E or Y88D claim 1 , and the amino acid substitution at Y89 is Y89E or Y89D.34-. (canceled)5. The polypeptide of claim 1 , wherein the engineered p27 comprises a sequence having at least 90% sequence identity to the sequence of SEQ ID NO: 1.716-. (canceled)17. The polypeptide of claim 6 , wherein the engineered p27 comprises a sequence having at least 90% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 6 claim 6 , 4 claim 6 , 12 claim 6 , 10 claim 6 , 15 claim 6 , 13 claim 6 , 21 claim 6 , 19 claim 6 , 27 claim 6 , 25 claim 6 , 30 claim 6 , 28 claim 6 , 33 claim 6 , 31 claim 6 , 36 claim 6 , and 34.1824-. (canceled)25. A trimeric protein complex comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(i) the polypeptide of , or a phosphorylated, wild-type p27 or a fragment thereof;'}(ii) a Cdk4 or a variant thereof, or a Cdk6 or a variant thereof; and(iii) a CycD or a variant thereof,wherein the Cdk4 or the variant thereof or the Cdk6 or the variant ...

ASSAYS FOR DETECTING MODIFIED COMPOUNDS

Provided are methods and compositions which are useful for separating, isolating, detecting, and quantifying compounds of interest which have been modified chemically, enzymatically or catalytically from other compounds which have not been so modified. The modifications may take the form of functional groups which are gained, lost or retained by the compounds of interest. 1100-. (canceled)101. A reagent for determining the presence of an enzyme that adds or removes a charged functional group to an uncharged moiety on a substrate , the reagent comprising the substrate to which a first member and a second member of an energy transfer pair are covalently bound at a distance such that a FRET interaction does not occur unless the charged functional group is present ,wherein the second member of the energy transfer pair has a charge opposite to the charge of the functional group, andwherein the presence of the charged functional group to the substrate causes noncovalent binding of the second member of the energy transfer pair to the functional group, bringing the first member and the second member of the energy transfer pair close enough so that a FRET interaction occurs.102. The reagent of claim 101 , wherein the functional group is negative charged.103. The reagent of claim 102 , wherein the negatively charged functional group is a phosphate claim 102 , carboxyl claim 102 , sulfate claim 102 , nitro claim 102 , or acyl group.104. The reagent of claim 101 , wherein the second member of the energy transfer pair further comprises a chelator that binds a metal ion that binds to the functional group.105. The reagent of claim 104 , wherein the functional group is a phosphate.106. A reagent for determining the presence of an enzyme that adds or removes an uncharged functional group to a charged moiety on a substrate claim 104 , the reagent comprising the substrate to which a first member and a second member of an energy transfer pair are covalently bound at a distance such ...

Heat stable mutants of starch biosynthesis enzymes

The subject invention pertains to novel mutant polynucleotide molecules that encode enzymes that have increased heat stability. These polynucleotides, when expressed in plants, result in increased yield in plants grown under conditions of heat stress. The polynucleotide molecules of the subject invention encode maize endosperm ADP glucose pyrophosphorylase (AGP) and soluble starch synthase (SSS) enzyme activities. Plants and plant tissue bred to contain, or transformed with, the mutant polynucleotides, and expressing the polypeptides encoded by the polynucleotides, are also contemplated by the present invention. The subject invention also concerns methods for isolating polynucleotides and polypeptides contemplated within the scope of the invention. Methods for increasing yield in plants grown under conditions of heat stress are also provided.

Treatment of Hearing Loss by Inhibition of Casein Kinase 1

Methods for treating hearing loss that include administering an inhibitor, e.g., a small molecule inhibitor, of casein kinase 1, preferably in combination with a treatment that stimulates Atoh1 gene expression, e.g., a gamma-secretase inhibitor, an Atoh1 stimulatory compound, or a GSK-3-beta inhibitor. 1. A method of treating a subject who has or is at risk of developing hearing loss or vestibular dysfunction , comprising administering to the subject one or more inhibitory nucleic acids that target casein kinase 1 (CK1) epsilon and/or CK1 delta , and one or more compounds that stimulate Atoh1 gene expression.2. The method of claim 1 , wherein the subject has or is at risk for developing sensorineural hearing loss claim 1 , auditory neuropathy claim 1 , or both.3. The method of claim 1 , wherein the subject has or is at risk for developing a vestibular dysfunction that results in dizziness claim 1 , imbalance claim 1 , or vertigo.4. The method of claim 1 , wherein the one or more inhibitory nucleic acids that target CK1 epsilon and/or CK1 delta claim 1 , and/or the one or more compounds that stimulate Atoh1 gene expression is administered systemically.5. The method of claim 1 , wherein the one or more inhibitory nucleic acids that target CK1 epsilon and/or CK1 delta claim 1 , and/or the one or more compounds that stimulate Atoh1 gene expression is administered locally to the inner ear.6. The method of claim 1 , wherein the one or more inhibitory nucleic acids that target CK1 epsilon and/or CK1 delta is shRNA or siRNA.7. The method of claim 1 , wherein the method comprises administering to the subject one or more inhibitory nucleic acids that target CK1 epsilon.8. The method of claim 1 , wherein the method comprises administering to the subject one or more inhibitory nucleic acids that target CK1 delta.9. The method of claim 1 , wherein the one or more compounds that stimulate Atoh1 gene expression comprises a glycogen synthase kinase 3 beta (GSK-3-beta) inhibitor.10. ...

Contemporaneous, heterogeneously-oriented, multi-targeted therapeutic modification and/or modulation of disease by administration of sulfur-containing, amino acid-specific small molecules

The present invention discloses and claims novel pharmaceutical compositions, methods, and kits used for the contemporaneous, heterogeneously-oriented, multi-targeted therapeutic modification and/or modulation of cellular metabolic anomalies or other undesirable physiological conditions, including cancer, where the normal cellular biochemical function and/or the expression levels of various proteins/enzymes (i.e., the target molecules) are abnormal and must be modified and/or modulated in order to treat these metabolic anomalies or other undesirable physiological conditions, including cancer. The aforementioned target molecules, by way of non-limiting example, include: anaplastic lymphoma kinase (ALK), mesenchymal epithelial transition (MET) kinase, the receptor tyrosine kinase (ROS1), epidermal growth factor receptor (EGFR), peroxiredoxin (Prx), excision repair cross-complementing protein 1 (ERCC1), insulin growth factor 1 receptor (IGF1R), ribonucleotide reductase (RNR), tubulin, farnesyltransferase, and various other classes of proteins/enzymes. Additionally, the present invention discloses and claims methods and kits for (a) the selection of subjects for treatment; (b) the determination of the most effective medicinal agent(s) to be administered in combination with the administration of the sulfur-containing, amino acid-specific small molecules of the present invention; (c) the dosage of the medicinal agent(s) to be administered; (d) the determination of the length and/or number of treatment cycles; (e) the adjustment of the specific medicinal agent(s) used and the dosage administered during treatment; and/or (f) ascertaining the potential treatment responsiveness of the specific disease to the medicinal agents (s) selected for administration to a subject suffering from one or more types of: (i) cancer or (ii) metabolic anomalies or other undesirable physiological conditions by quantitatively determining the level of the abnormal biochemical activity and/or ...

Kinase activity detection methods

The invention provides cellular assays for detecting the activity of one or more kinase from multiple conditions simultaneously, by encoding biochemically identical substrates with isotope labels that enable them to be distinguished in pooled samples by mass spectrometry.

BIOMARKER FOR PARKINSON'S DISEASE AND USE THEREOF

A antibody has as a target molecule a ubiquitin protein comprising a phosphorylated serine residue at position 65. In addition, a method is provided for specifically detecting Parkinson's disease at an early stage, in which a target molecule is a ubiquitin protein comprising a phosphorylated serine residue at position 65, a pharmaceutical composition for definitively treating or preventing Parkinson's disease, and a method for screening for the pharmaceutical composition. 110-. (canceled)11. A ubiquitin protein comprising a phosphorylated serine residue at position 65 (SEQ ID NO: 1) , or a ubiquitin protein consisting of an amino acid sequence comprising a substitution , deletion , insertion and/or addition of 1 to 3 amino acids with respect to the amino acid sequence shown in SEQ ID NO: 1 , on condition that the phosphorylated serine residue at position 65 is conserved.12. A phosphorylation-mimicking form of ubiquitin , in which the serine residue at position 65 is substituted with an aspartic acid residue (SEQ ID NO: 2) , or a phosphorylation-mimicking form of ubiquitin consisting of an amino acid sequence comprising a substitution , deletion , insertion and/or addition of 1 to 3 amino acids with respect to the amino acid sequence shown in SEQ ID NO: 2 , on condition that the aspartic acid residue at position 65 is conserved.13. A method for testing for Parkinson's disease , comprising a step of detecting or quantifying a ubiquitin protein comprising a phosphorylated serine residue at position 65 in a sample isolated from a subject.14. The method according to claim 13 , wherein the step of detecting or quantifying is carried out by an immunological technique.15. A biomarker for detecting Parkinson's disease claim 13 , which consists of a ubiquitin protein comprising a phosphorylated serine residue at position 65.16. An antibody having an ability to specifically bind to a ubiquitin protein comprising a phosphorylated serine residue at position 65.17. The antibody ...

AMINO ACID-SENSING DIGUANYLATE CYCLASE AND METHODS OF USE

Provided are SpdE polypeptides and variants and nucleic acids encoding the SpdE polypeptides and variants. Also provided are vectors including one or more nucleic acids encoding a SpdE polypeptide or variant and cells including a nucleic acid encoding the SpdE polypeptide or variant, as well as cells expressing a SpdE polypeptide or variant and compositions including such cells and a pharmaceutically acceptable carrier. Finally, methods of detecting presence and/or amount of one or more amino acids in a sample are provided. The methods include contacting the sample with a SpdE protein, measuring diguanylate cyclase activity of the SpdE protein; and comparing the diguanylate cyclase activity of the SpdE protein to a control. The methods can utilize isolated SpdE protein or a cell expressing a SpdE protein. 1. A method of detecting presence or amount of one or more amino acids in a sample , comprising:contacting the sample with a SpdE protein;measuring diguanylate cyclase activity of the SpdE protein; andcomparing the diguanylate cyclase activity of the SpdE protein to a control, wherein a decrease in diguanylate cyclase activity compared to the control indicates the presence or amount of one or more amino acids in the sample.2. The method of claim 1 , wherein the one or more amino acids are one or more of proline claim 1 , valine claim 1 , isoleucine claim 1 , leucine claim 1 , alanine claim 1 , methionine claim 1 , or threonine.3. The method of claim 1 , wherein measuring the diguanylate cyclase activity of the SpdE protein comprises measuring an amount of cyclic-di-GMP or pyrophosphate.4. The method of claim 1 , wherein contacting the sample with the SpdE protein comprises contacting the sample with a cell expressing the SpdE protein.5. The method of claim 4 , wherein measuring the diguanylate cyclase activity of the SpdE protein comprises measuring the motility of the cell expressing the SpdE protein and wherein an increase in the motility of the cell compared to ...

METHODS AND COMPOSITIONS FOR THE DIAGNOSIS AND FOR THE TREATMENT OF ADRENOLEUKODYSTROPHY

The present invention is directed to a diagnostic method for adrenoleukodystrophy in a subject based on the determination of the levels of different markers. The invention also provides a method for monitoring the progression of an adrenoleukodystrophy, a method for monitoring the effect of an adrenoleukodystrophy therapy and fingolimod, an analogue, metabolite or derivative thereof, or a pharmaceutically acceptable salt thereof, for use in the treatment and/or prevention of anadrenoleukodystrophy. 1. (canceled)4. The method according to wherein the therapy comprises an antioxidant compound.5. The method according to wherein the antioxidant compound is a combination of N-acetylcysteine claim 4 , lipoic acid and vitamin E.6. The method according to wherein the adrenoleukodystrophy occurs without inflammatory demyelination.7. The method according to wherein the sample is a sample containing peripheral mononuclear cells.8. The method according to wherein the adrenoleukodystrophy is selected from the group consisting of adult adrenomyeloneuropathy (AMN) claim 2 , cerebral adrenomyeloneuropathy (cAMN) and the childhood variant of drenoleukodystrophy (cALD).920-. (canceled)21. A method for the treatment and/or prevention of an adrenoleukodystrophy in a subject in need thereof comprising the administration of an inhibitor of sphingosine-1-phosphate receptor 1 or a pharmaceutical composition comprising said inhibitor.22. The method according to claim 21 , wherein said inhibitor is fingolimod claim 21 , an analogue claim 21 , metabolite or derivative thereof claim 21 , or a pharmaceutically acceptable salt thereof.23. The method according to wherein the fingolimod analogue is siponimod.24. The method according to wherein the adrenoleukodystrophy is selected from the group consisting of adult adrenomyeloneuropathy (AMN) claim 21 , cerebral adrenomyeloneuropathy (cAMN) and the childhood variant of adrenoleukodystrophy (cALD).25. The method according to wherein the ...

USE OF PHOSPHO-AKT AS A BIOMARKER OF DRUG RESPONSE

Use of phospho-Akt as a biomarker for predicting the response, such as resistance, to a compound, wherein phospho-Akt is Akt that has been phosphorylated on one or more residues, with the proviso that fir Akt1, Akt2, and Akt3 the designation phospho-Akt is used to indicate phosphorylation at a site other than T308, T309 or T305 respectively, wherein the compound is a compound of general formula (I) wherein R represents phenyl, thienyl or pyridinyl wherein phenyl is optionally substituted by one or two substituents independently selected from alkyl, halo-lower alkyl, hydroxy-lower alkyl, lower alkoxy-lower alkyl, acyloxy-lower alkyl, phenyl, hydroxy, lower alkoxy, hydroxy-lower alkoxy, lower alkoxy-lower alkoxy, phenyl-lower alkoxy, lower alkylcarbonyloxy, amino, monoalkylamino, dialkylamino, lower alkoxycarbonylamino, lower alkylcarbonylamino, substituted amino wherein the two substituents on nitrogen form together with the nitrogen heterocyclyl, lower alkylcarbonyl, carboxy, lower alkoxycarbonyl, cyano, halogen, and nitro; and wherein two adjacent substituents are methylenedioxy; and wherein pyridinyl is optionally substituted by lower alkoxy, amino or halogen; X represents a group C═Y, wherein Y stands for oxygen or nitrogen substituted by hydroxy or lower alkoxy; Rrepresents hydrogen, lower alkylcarbonyl, hydroxy-lower alkyl or cyano-lower alkyl; R, Rand Rrepresent hydrogen; Rand R, independently of each other, represent hydrogen, lower alkyl or lower alkoxy, or Rand Rtogether represent methylenedioxy, and pharmaceutically acceptable derivatives thereof; or wherein R represents phenyl or pyridinyl wherein phenyl is optionally substituted by one or two substituents independently selected from alkyl, halo-lower alkyl, hydroxy-lower alkyl, lower alkoxy-lower alkyl, acyloxy-lower alkyl, phenyl, hydroxy, lower alkoxy, hydroxy-lower alkoxy, lower alkoxy-lower alkoxy, phenyl-lower alkoxy, lower alkylcarbonyloxy, amino, monoalkylamino, dialkylamino, lower ...

ASSAYS FOR DETECTING MODIFIED COMPOUNDS

Provided are methods and compositions which are useful for separating, isolating, detecting, and quantifying compounds of interest which have been modified chemically, enzymatically or catalytically from other compounds which have not been so modified. The modifications may take the form of functional groups which are gained, lost or retained by the compounds of interest. 1. A method for detecting or quantifying an enzyme capable of adding a functional group to a compound in a sample , comprising the steps of: (i) a compound which can gain a functional group from an enzyme, the compound comprising at least one non-radioactive signaling moiety;', '(ii) a sample suspected of containing an enzyme that could add a functional group to the compound; and, '(a) providing(b) forming a mixture comprising the compound and the sample; and(c) separating any compound that has gained the functional group; and(d) detecting or quantifying the non-radioactive signaling moiety of any separated compounds having the functional group, thereby detecting or quantifying the enzyme in the sample.2. The method of claim 1 , wherein the enzyme is selected from the group comprising kinases claim 1 , phosphatases claim 1 , sulfatases claim 1 , sufotransferases claim 1 , acetyltransferases claim 1 , deacetylases claim 1 , methylases claim 1 , demethylases claim 1 , carboxylases claim 1 , decarboxylases claim 1 , glycosylases claim 1 , amidases claim 1 , deamidases claim 1 , aminases and deaminases.3. The method of claim 1 , wherein the enzyme is selected from the group comprising kinases claim 1 , phosphatases claim 1 , or sulfotransferases.4. The method of claim 1 , wherein the enzyme is sphingosine kinase 1 or sphingosine kinase 2.5. The method of claim 1 , wherein the enzyme is sphingosine kinase 1.6. The method of claim 1 , wherein the enzyme is sphingosine kinase 2.7. The method of claim 1 , wherein the non-radioactive signaling moiety comprises a fluorescent compound claim 1 , a ...

STAT5a AND ITS FUNCTIONAL TUMOR SUPPRESSOR ANALOGS FOR TREATMENT OF MALIGNANCIES EXPRESSING NPM/ALK AND OTHER ONCOGENIC KINASES

The invention provides methods of inhibiting epigenetic gene silencing in a cell expressing NPM/ALK or decreasing NPM/ALK content in a cell, by contacting a cell with an agent capable of increasing the concentration of Stat5a protein or its functional analog. Further, the invention provides a method of treating malignancies expressing oncogenic kinase by administering to a patient affected with a malignancy an agent capable of increasing the concentration of Stat5a protein or its epigenetically silenced functional tumor suppressor analog in a malignant cell. Finally, it provides a method to diagnose malignancy and monitor patient's response to therapy by analysis of the degree of DNA methylation of the gene encoding for Stat5a or its analog, their mRNA, or protein. 110.-. (canceled)11. A method of decreasing NPM/ALK , or another oncogenic -kinase content in a cell , comprising the step of contacting a cell expressing NPM/ALK or another oncogenic -kinase , with an effective amount of an agent capable of increasing the amount of Stat5a or its functional analog , thereby down-regulating the kinase gene expression in the cell.12. The method of claim 11 , whereby the agent is Stat5a protein claim 11 , a Stat5a mRNA claim 11 , Stat5a DNA or a combination thereof.13. The method of claim 12 , further comprising a siRNA claim 12 , a polyamide claim 12 , a triple-helix-forming agent claim 12 , an antisense RNA claim 12 , a synthetic peptide nucleic acids (PNAs) claim 12 , an agRNA claim 12 , a LNA/DNA copolymer claim 12 , a small molecule chemical compound claim 12 , or a combination thereof claim 12 , specific against a NPM/ALK claim 12 , another form of ALK claim 12 , another chimeric tyrosine kinase claim 12 , or another oncogenic kinase.14. A method of treating malignancies expressing an ALK chimeric tyrosine kinase in a subject claim 12 , comprising the step of administering to the subject a composition comprising a therapeutically effective amount of a small molecule ...

METHOD TO DETERMINE RESPONSIVENESS OF CANCER TO EPIDERMAL GROWTH FACTOR RECEPTOR TARGETING TREATMENTS

Disclosed herein are methods and reagents for determining the responsiveness of cancer to an epidermal growth factor receptor (EGFR) targeting treatment. The detection of these mutations will allow for the administration of gefitinib, erlotinib and other tyrosine kinase inhibitors to those patients most likely to respond to the drug. 1. An assay comprising: i) a mutation in exon 18 that results in a substitution of cysteine for glycine at position 719 (G719C) or in a substitution of serine for glycine at position 719 (G719S) or in a substitution of an alanine for glycine at position 719 (G719A) of SEQ ID NO: 512;', 'ii) an in-frame deletion in exon 19 that results in a deletion of at least amino acids leucine, arginine, glutamic acid and alanine at codons 747, 748, 749, and 750 of SEQ ID NO: 512;', 'iii) a mutation in exon 20 that results in a insertion of amino acids asparagine, proline and glycine between position 770 and 771 (D770_N771insNPG), or in an insertion of amino acids serine, valine and aspartic acid between position 770 and 771 (D770_N771insSVD), or in an insertion of amino acid valine between position 772 and 773 (P772_H773insV), or in a substitution at position 790 of SEQ ID NO: 512; and', 'iv) a mutation in exon 21 that results in an amino acid substitution of arginine for leucine at position 858 (L858R) or of glutamine for leucine at position 861 (L861Q) of SEQ ID NO: 512;, '(a) adding primers specific for at least one of the following nucleotide variances in an epidermal growth factor receptor (EGFR) gene, where the nucleotide variance is selected fromto a biological sample obtained from the blood of a human patient afflicted with non-small cell lung cancer;(b) performing an amplification step by polymerase chain reaction (PCR) wherein the PCR is allele-specific amplification for at least one of the nucleotide variances; and(c) detecting whether at least one of the above-described variances is present.2. The assay of claim 1 , wherein the allele- ...

CRYSTALLINE FORM OF (S)-N-(5-((R)-2-(2,5-DIFLUOROPHENYL)-PYRROLIDIN-1-YL)-PYRAZOLO[1,5-A]PYRIMIDIN-3-YL)-3-HYDROXYPYRROLIDINE-1-CARBOXAMIDE HYDROGEN SULFATE

A novel crystalline form of (S)—N-(5-((R)-2-(2,5-difluorophenyl)pyrrolidin-1-yl)-pyrazolo[1,5-a]pyrimidin-3-yl)-3-hydroxypyrrolidine-1-carboxamide, pharmaceutical compositions containing said crystalline form and the use of said crystalline form in the treatment of pain, cancer, inflammation, neurodegenerative disease or Trypanosoma cruzi infection are disclosed. In some embodiments, the novel crystalline form comprises a stable polymorph of (S)—N-(5-((R)-2-(2,5-difluorophenyl)pyrrolidin-1-yl)-pyrazolo[1,5-a]pyrimidin-3-yl)-3-hydroxypyrrolidine-1-carboxamide hydrogen sulfate. The present invention is further directed to a process for the preparation of the novel crystalline form. 2. The method of claim 1 , wherein the crystalline form is characterized by having XRPD diffraction peaks (2θ degrees) at 10.7±0.2 claim 1 , 18.4±0.2 claim 1 , 20.7±0.2 claim 1 , 23.1±0.2 claim 1 , and 24.0±0.2.3. The method of claim 1 , wherein the crystalline form is characterized by having XRPD diffraction peaks (2θ degrees) at 10.7±0.2 claim 1 , 18.4±0.2 claim 1 , 19.2±0.2 claim 1 , 20.2±0.2 claim 1 , 20.7±0.2 claim 1 , 21.5±0.2 claim 1 , 23.1±0.2 claim 1 , and 24.0±0.2.4. The method of claim 1 , wherein the crystalline form is characterized by having XRPD diffraction peaks (2θ degrees) at 10.7±0.2 claim 1 , 15.3±0.2 claim 1 , 16.5±0.2 claim 1 , 18.4±0.2 claim 1 , 19.2±0.2 claim 1 , 19.9±0.2 claim 1 , 20.2±0.2 claim 1 , 20.7±0.2 claim 1 , 21.5±0.2 claim 1 , 22.1±0.2 claim 1 , 23.1±0.2 claim 1 , 24.0±0.2. 24.4±0.2 claim 1 , 25.6±0.2 claim 1 , 26.5±0.2 claim 1 , 27.6±0.2 claim 1 , 28.2±0.2 claim 1 , 28.7±0.2 claim 1 , 30.8±0.2 claim 1 , and 38.5±0.2.5. The method of claim 1 , wherein the crystalline form exhibits an onset to maximum of about 193° C. to about 205° C. claim 1 , as measured by differential scanning calorimetry.6. The method of claim 1 , wherein the crystalline form exhibits a heat of melting of about 2.415 mW claim 1 , as measured by differential scanning calorimetry.7. The ...

BIOLOGICAL SAMPLE AND BIOLOGICAL INSTRUMENT CLEANLINESS MEASUREMENT KIT AND METHOD

Provided are a kit and a method for measuring cleanliness of a bio-related sample and a bio-related instrument, which is not susceptible to ATP-degrading activity. Provided are a method for measuring cleanliness of a bio-related sample or a bio-related instrument and a kit therefor, comprising using an enzyme that catalyzes a reaction that produces ATP from ADP, luciferin, luciferase, and a metal salt. Also provided are a method further comprising using pyruvate orthophosphate dikinase (PPDK), adenylate kinase (ADK), or pyruvate-water dikinase (PWDK) and a kit further comprising PPDK, ADK, or PWDK. Further provided are a method for measuring cleanliness of a bio-related sample or a bio-related instrument and a kit therefor, comprising using an enzyme that catalyzes a reaction that produces ATP from AMP, an enzyme that catalyzes a reaction that produces AMP from ADP, luciferin, luciferase, and a metal salt. 1. A kit for measuring cleanliness of a blood-related sample or a blood-related instrument , comprising an enzyme that catalyzes a reaction that produces ATP from ADP , luciferin , luciferase and a metal salt.2. A kit for measuring cleanliness of a bio-related sample or a bio-related instrument , comprising an enzyme that catalyzes a reaction that produces ATP from ADP , luciferin , luciferase , and a metal salt.3. The kit according to claim 1 , wherein the enzyme that catalyzes a reaction that produces ATP from ADP is selected from the group consisting of pyruvate kinase (PK) claim 1 , acetate kinase (AK) claim 1 , creatine kinase (CK) claim 1 , polyphosphate kinase (PPK) claim 1 , hexokinase claim 1 , glucokinase claim 1 , glycerol kinase claim 1 , fructokinase claim 1 , phosphofructokinase claim 1 , riboflavin kinase claim 1 , and fructose-bisphosphatase.4. The kit according to claim 1 , further comprising an enzyme that catalyzes a reaction that produces ADP or ATP from AMP.5. The kit according to claim 4 , wherein the enzyme that catalyzes a reaction that ...

METHODS TO IDENTIFY AND TREAT SUBJECTS HAVING CORTICOSTEROID-RESISTANT INFLAMMATORY DISEASES

The present invention is directed toward novel methods to identify as well as to treat a subject having an inflammatory disease resistant to corticosteroids. 144.-. (canceled)45. A method to identify a subject having an inflammatory disease resistant to corticosteroid treatment comprising determining the level of phosphorylated MSK1 (p-MSKI) in a biological sample from the subject.46. The method of claim 45 , wherein the step of determining the subject's p-MSK1 level comprises determining the subject's p-MSK1 level claim 45 , wherein an elevated p-MSK1 level in the subject as compared to a control p-MSK1 level indentifies the subject as having an inflammatory disease resistant to corticosteroid treatment.47. The method of wherein the biological sample is selected from the group consisting of urine claim 45 , blood claim 45 , sputum claim 45 , bronchoalveolar lavage (BAL) cells and peripheral blood mononuclear cells (PBMCs).48. The method of claim 45 , wherein the inflammatory disease is selected from the group consisting of inflammatory bowel disease claim 45 , allergic rhinitis claim 45 , sinusitis claim 45 , atopic dermatitis claim 45 , psoriasis claim 45 , arthritis claim 45 , inflammatory lung disease claim 45 , interstitial lung disease claim 45 , sarcoidosis claim 45 , and an autoimmune inflammatory disease.49. (canceled)50. (canceled)51. The method of claim 48 , wherein the inflammatory lung disease is triggered by the subject's exposure to environmental conditions.52. (canceled)53. The method of claim 48 , wherein the inflammatory lung disease is asthma.54. The method of claim 45 , wherein the subject is being administered a non-steroidal anti-inflammatory drug.55. (canceled)56. A method for diagnosing and treating an inflammatory disease resistant to corticosteroid treatment in a subject comprising analyzing a subject sample for the presence of p-MSK1 claim 45 , wherein the subject is diagnosed as having an inflammatory disease resistant to corticosteroid ...

Camkk-beta as a target for treating cancer

Provided herein are compounds, compositions, including pharmaceutical compositions, having anti-cancer activity. Also provided are methods for diagnosing, detecting, and treating cancer in a subject, as well as a method for evaluating cancer stage in a subject, wherein the methods include determining the amount of a Ca 2+ /calmodulin dependent kinase kinase (CaMKK) in a sample. Further provided are methods of screening and identifying a compound that inhibits CaMKK.

IDENTIFICATION OF MOLECULAR PATHWAYS AND METHODS OF USE THEREOF FOR TREATING RETINAL NEURODEGENERATION AND OTHER NEURODEGENERATIVE DISORDERS

Drug targets, pathways, kits and methods for treating conditions related to neurodegeneration or ocular disease, are disclosed. 1. A method for inhibiting or preventing retinal ganglion cell injury or death , the method comprising:contacting the retinal ganglion cell with a small molecule that modulates protein kinase expression or activity, wherein the protein kinase is selected from the group consisting of MAP3K12, MAP3K13, MAP3K14, MAP2K7, MAP2K4, LYN, PLK3, PFKP MARK3, MARK2, TAOK1, IKBKB, BRSK2, PBK, PRKCH, TESK1, Csnk1e, Oxsr1, Tgfbr2, Mapk10, PFTK1, ERN2, AK2, HSPB8, FRAP1, DGUOK, ERN1, STK32B, PIK3C2G, BCR, DYRK1A, DYRK1B, PNCK, EIF2AK1, PKD2L1, NRK, Endothelin Receptor Type B, and TLK2.2. The method of claim 1 , wherein the modulation of protein kinase expression or activity is an inhibition of expression or activity.3. The method of claim 1 , wherein the modulation of protein kinase expression or activity is an increase of expression or activity.4. The method of claim 1 , wherein the small molecule is a short interfering RNA (siRNA).5. The method of claim 4 , wherein the siRNA is a siRNA targeting MAP3K12 claim 4 , MAP3K13 claim 4 , MAP2K4 claim 4 , or MAP2K7.6. The method of claim 1 , wherein the small molecule is selected from the group consisting of CEP-1347 and CEP-11004.7. The method of claim 1 , wherein the contacting of the retinal ganglion cell with a small molecule is performed ex vivo or in vivo.8. The method of claim 7 , wherein the method further comprises grafting or implanting the retinal ganglion cell into a subject after contacting the cell with a small molecule.9. The method of claim 8 , wherein the retinal ganglion cell is in a pharmaceutically acceptable carrier.10. The method of claim 1 , wherein the inhibiting or preventing retinal ganglion cell injury or death treats or prevents an ocular neurodegenerative disease.11. The method of claim 10 , wherein the ocular neurodegenerative disease is selected from the group consisting of ...

COMPOSITIONS INCLUDING TRICIRIBINE AND ONE OR MORE PLATINUM COMPOUNDS AND METHODS OF USE THEREOF

This application encompasses combination therapies including triciribine and related compounds and one or more platinum compounds and compositions with reduced toxicity for the treatment and prevention of tumors, cancer, and other disorders associated with abnormal cell proliferation. 125-. (canceled)27. The composition of claim 26 , wherein the compound of formula I is triciribine.28. The composition of claim 26 , wherein the compound of formula I is triciribine phosphate.29. The composition of claim 26 , wherein the compound of formula I is triciribine phosphonate.30. The composition of claim 26 , wherein the compound of formula I is present in a dose amount of at least 20 mg/m.31. The composition of claim 26 , wherein the compound of formula I is present in an amount of at least 10 mg/m.32. The method of claim 26 , wherein the administration is parenteral administration.33. The method of claim 32 , wherein the parenteral administration is intravenous administration.34. The method of claim 26 , wherein the administration is oral administration.35. The method of claim 26 , suitable for intravenous administration.36. The method of claim 26 , wherein the one or more platinum compounds is present in a dose about 0.1 mg/mto about 200 mg/m.37. The method of claim 26 , wherein the one or more platinum compounds is present in a dose about 1 mg/mto about 150 mg/m.38. The method of claim 26 , wherein the one or more platinum compounds is present in a dose about 10 mg/mto about 100 mg/m.39. The composition of claim 26 , wherein the one or more platinum compounds is present in a dose about 25 mg/mto about 50 mg/m.40. The method of claim 26 , wherein the administration of a compound of formula I and the one or more platinum compounds is concurrently administered.41. The method of claim 26 , wherein the administration of a compound of formula I is followed by the administration of the one or more platinum compounds.42. The method of claim 26 , wherein the administration of the ...

CLASSIFICATION OF KINASE INHIBITORS USING NONLINEAR OPTICAL TECHNIQUES

A method is disclosed for classifying and distinguishing between type I and type II kinase inhibitors. The method involves the use of non-linear optical techniques, in particular second-harmonic generation (SHG) to identify conformational changes in kinase proteins obtained from known type I or type II inhibitors. The method further involves deducing the manner of binding of unknown inhibitors by comparison with the signal changes produced by known ligands. The method is also applied to comparing the conformational changes induced by the binding of generic and branded kinase inhibitor drugs to a target kinase. 1. A method for classifying a kinase inhibitor as a type I or type II kinase inhibitor based on a conformational change that the kinase inhibitor induces in the structure of a kinase labeled with a second harmonic-active label , wherein the label has a net orientation at an interface , the method comprising:a. contacting the kinase with a kinase inhibitor, wherein the kinase specifically interacts with said kinase inhibitor;b. detecting an interaction between the kinase and said kinase inhibitor by measuring a first signal or signal change generated by the second harmonic-active label using a surface-selective technique, wherein the first signal or signal change indicates a conformational change in the structure of the kinase that is specific for the kinase inhibitor; andc. classifying the kinase inhibitor as a type I or type II kinase inhibitor by comparing the first signal or signal change of (b) with a second signal or signal change detected by an interaction between the kinase and a known type I or type II inhibitor of the kinase, wherein the second signal or signal change indicates a conformational change in the structure of the kinase that is specific for the known type I or type II inhibitor of the kinase.27-. (canceled)8. The method of any of claim 1 , wherein the kinase comprises an affinity tag.9. The method of claim 1 , wherein the conformational ...

COMPOSITIONS AND METHODS RELATING TO FUSION PROTEIN BIOMARKERS

The present invention provides fusion proteins as biomarkers specific for chromosomal translocation-based conditions (e.g., cancer), related methods for detecting fusion protein biomarkers associated with chromosomal translocation-based conditions, related methods for quantifying amount of fusion protein expression, and related methods for diagnosing chromosomal translocation-based conditions through detection of such fusion protein biomarkers. Such fusion protein biomarkers and related methods additionally find use in research settings. 1. A method for detecting the presence of one or more fusion proteins within a biological sample , comprising: a biological sample, wherein said biological sample is a blood sample and/or a tissue sample,', wherein said labeled peptides within each distinct group comprise peptides having one isotopically labeled amino acid residue and peptides having two isotopically labeled amino acid residues,', 'wherein said amino acid sequence of said labeled peptides within each distinct group is identical with an amino acid sequence of a particular fusion protein peptide fragment generated through digestion of said particular fusion protein with a protease,, 'one or more distinct groups of labeled peptides, wherein each distinct group of labeled peptides is specific for a particular fusion protein to be detected, wherein the labeled peptides within each distinct group have labeled peptides having an amino acid sequence that spans the fusion junction of said particular fusion protein encoded by a fusion of a first nucleic acid to a second nucleic acid,'}, 'one or more proteases known to generate upon digestion of said two or more fusion proteins to be detected particular fusion protein peptide fragments,, 'a) providingb) digesting said biological sample with said one or more proteases resulting in generation of biological sample-based peptide fragments,c) combining said one or more distinct groups of labeled peptides with said generated ...

IMIDAZOPYRAZINE SYK INHIBITORS

Certain imidazopyrazines and pharmaceutical compositions thereof are provided herein. Methods of treating patients suffering from certain diseases and disorders responsive to the inhibition of Syk activity, which comprises administering to such patients an amount of at least one chemical entity effective to reduce signs or symptoms of the disease or disorder are provided. Also provided are methods for determining the presence or absence of Syk kinase in a sample. 152-. (canceled)54. A pharmaceutical composition comprising the compound of claim 53 , or a pharmaceutically acceptable salt thereof claim 53 , and at least one pharmaceutically acceptable vehicle chosen from carriers claim 53 , adjuvants claim 53 , and excipients.55. A method for treating a human having a disease responsive to the inhibition of Syk activity claim 53 , comprising administering to the human an effective amount of the compound according to claim 53 , or a pharmaceutically acceptable salt thereof.56. The method according to claim 55 , wherein an effective amount of the compound is administered to the human intravenously claim 55 , intramuscularly claim 55 , parenterally claim 55 , or orally.57. The method according to claim 55 , wherein the disease responsive to inhibition of Syk activity is cancer.58. The method according to claim 55 , wherein the disease responsive to inhibition of Syk activity is B-cell lymphoma or leukemia.59. The method according to claim 55 , wherein the disease responsive to inhibition of Syk activity is rheumatoid arthritis claim 55 , allergic rhinitis claim 55 , chronic obstructive pulmonary disease (COPD) claim 55 , adult respiratory distress syndrome (ARDs) claim 55 , an allergy-induced inflammatory disease claim 55 , multiple sclerosis claim 55 , an autoimmune disease claim 55 , an inflammatory disease claim 55 , an acute inflammatory reaction claim 55 , an allergic disorder claim 55 , polycystic kidney disease claim 55 , B-cell lymphoma claim 55 , Hodgkin's ...

KIT AND METHOD

The present invention relates to a kit comprising a DNA-dependant DNA polymerase and at least one natural deoxynucleoside and to a kit comprising a DNA-dependent DNA polymerase and a detection system comprising a DNA template molecule, a DNA primer molecule, and a fluorescent moiety capable of being displaced from, or bound to, dsDNA synthesized by said DNA-dependent polymerase. It further relates to A method for measuring deoxynucleoside kinase activity in a sample characterized by; (i) contacting the sample, in a container, with a reaction mix comprising a DNA-dependent DNA polymerase, at least one deoxynucleoside and a detection system comprising a DNA template molecule, a DNA primer molecule, and a fluorescent moiety capable of being incorporated in, displaced from, or bound to dsDNA synthesized by said DNA dependent polymerase; (ii) incubating said container; (iii) measuring the signal from the fluorescent moiety; and correlating the signal from the fluorescent moiety to the deoxynucleoside kinase activity in the sample. 1. A kit comprising a DNA-dependant DNA polymerase and at least one natural deoxynucleoside.2. A kit according to claim 1 , further comprising a detection system comprising a DNA template molecule claim 1 , a DNA primer molecule claim 1 , and a fluorescent moiety capable of being displaced from claim 1 , or bound to claim 1 , dsDNA synthesized by said DNA-dependent polymerase.3. A kit according to claim 1 , further comprising at least a phosphotransferase.4. A kit according to claim 1 , further comprising at least one modified deoxynucleoside.5. A kit according to claim 1 , further comprising at least one modified deoxynucleoside chosen from deoxythymidine and/or deoxycytidine.6. A kit according to claim 1 , further comprising at least one modified deoxynucleoside chosen from modified thymidine and modified cytidine claim 1 , such as BrdU and/or IdU.7. A kit according to claim 2 , wherein the fluorescent moiety is coupled to a single stranded ...

Protein Kinase C Inhibitors and Uses Thereof

This disclosure concerns compounds which are useful as inhibitors of protein kinase C (PKC) and are thus useful for treating a variety of diseases and disorders that are mediated or sustained through the activity of PKC. This disclosure also relates to pharmaceutical compositions comprising these compounds, methods of using these compounds in the treatment of various diseases and disorders, processes for preparing these compounds and intermediates useful in these processes. 189.-. (canceled)91. The method of claim 90 , wherein the lipase performs enzyme-catalyzed acylation of one of the stereoisomers in the racemic mixture.92. The method of claim 90 , further comprising after the contacting claim 90 , separating a stereoisomer from the racemic mixture.93. The method of claim 92 , wherein the separating comprising separating the stereoisomer by chromatography.94. The method of claim 92 , wherein the separated stereoisomer comprises an acyl group.95. The method of claim 92 , wherein the separated stereoisomer comprises a hydroxyl group.96. The method of claim 95 , further comprising converting the hydroxyl group of the separated stereoisomer to an amino group.99. The method of claim 98 , wherein the reaction is run in a polar aprotic or polar protic solvent.100. The method of claim 98 , further comprising performing separation of isomers with chiral chromatography.101. The method of claim 98 , further comprising performing separation of isomers with a resolution technique.102. The method of claim 98 , wherein Aris aryl or substituted aryl.103. The method of claim 98 , wherein Aris heteroaryl or substituted heteroaryl. Pursuant to 35 U.S.C. §119(e), this application claims priority to the filing date of U.S. Provisional Application No. 61/620,232, filed Apr. 4, 2012, and U.S. Provisional Application No. 61/783,647, filed Mar. 14, 2013, the disclosures of each of which are herein incorporated by reference.Protein kinase C (“PKC”) is a key enzyme in signal transduction ...

TREATING COGNITIVE DECLINE AND OTHER NEURODEGENERATIVE CONDITIONS BY SELECTIVELY REMOVING SENESCENT CELLS FROM NEUROLOGICAL TISSUE

Methods are provided herein for selectively killing senescent cells and for treating senescence-associated diseases and disorders by administering a senolytic agent. Senescence-associated diseases and disorders treatable by the methods using the senolytic agents described herein include cardiovascular diseases and disorders associated with or caused by arteriosclerosis, such as atherosclerosis; idiopathic pulmonary fibrosis; chronic obstructive pulmonary disease; osteoarthritis; senescence-associated ophthalmic diseases and disorders; and senescence-associated dermatological diseases and disorders. 1. A method of treating a neurodegenerative disease or disorder that is not a cancer in a subject in need thereof , the method comprisingtreating the subject using a course of therapy with a pharmaceutical composition that contains a compound that constitutes a means for selectively inhibiting Bcl-2 or Bcl-xL,wherein the pharmaceutical composition is formulated and administered such that the compound contacts p16 positive senescent cells located in or around neurological tissue in the subject that are promoting symptoms of the neurodegenerative disease or disorder, thereby selectively removing such cells;wherein the course of therapy includes:(1) a treatment period during which the pharmaceutical composition is administered to the subject whereby the compound selectively removes senescent cells from the neurological tissue; followed by(2) a therapeutic period of at least two weeks, during which the pharmaceutical composition is not administered to the subject, and progression of the neurodegenerative disease or disorder is inhibited as a result of administration of the compound to the subject during the treatment period.2. The method of claim 1 , wherein the means for selectively inhibiting Bcl-2 or Bcl-xL is ABT-263 (Navitoclax) claim 1 , or a pharmaceutically acceptable salt thereof.3. The method of claim 1 , wherein the means for selectively inhibiting Bcl-2 or Bcl-xL ...

Novel Cell-Based Assay for Determining Activity in the Retinoblastoma Pathway

Disclosed are methods of determining activity of CDK4 and CDK6 variants upon exposure to CDK inhibitors, methods for determining activity of a Rb variant, methods for determining the activity of a p16 variant in a cell, and methods for determining the sensitivity of a CDK4 variant or a CDK6 variant to p16 in a cell. Stable cell lines for determining activity of CDK4 variants, CDK6 variants, Rb variants, and p16 variants are also disclosed. 14.-. (canceled)5. In another aspect , the present invention concerns a cell line comprising:a. a first construct comprising a first cDNA encoding E2F1 protein which is linked to Dp-1 protein and a first portion of a luciferase gene at its C-terminus; andb. a knockout of endogenous CDK4 and endogenous CDK6.6. In another aspect , the present invention concerns a cell line comprising:a. a first construct comprising a first cDNA encoding Dp-1 protein and a second cDNA encoding E2F1 protein which is linked to a first portion of a luciferase gene at its C-terminus; andb. a knockout of endogenous CDK4 and endogenous CDK6. This application claims the benefit of under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 62/587,107, filed Nov. 16, 2017 and Provisional Application No. 62/756,750, filed Nov. 7, 2018, the content of which are incorporated herein by reference in their entirety.The present invention generally relates to a cell-based assay useful for determining the activity of CDK4, CDK6, Rb, and p16, and their respective variants. The novel cell-based assay is useful in screening CDK4 and CDK6 inhibitors and predicting patient response to treatment with CDK targeted inhibitors.The cyclin-dependent kinases are serine/threonine protein kinases which are the driving force behind the cell cycle and cell proliferation. Specifically, CDK4 and CDK6 (CDK4/6) is a catalytic subunit of the protein kinase complex that is important for cell cycle G1 phase progression. The activity of this kinase is restricted to the G1-S phase, which is ...

TREATING COGNITIVE DECLINE AND OTHER NEURODEGENERATIVE CONDITIONS BY SELECTIVELY REMOVING SENESCENT CELLS FROM NEUROLOGICAL TISSUE

Methods are provided herein for selectively killing senescent cells and for treating senescence-associated diseases and disorders by administering a senolytic agent. Senescence-associated diseases and disorders treatable by the methods using the senolytic agents described herein include cardiovascular diseases and disorders associated with or caused by arteriosclerosis, such as atherosclerosis; idiopathic pulmonary fibrosis; chronic obstructive pulmonary disease; osteoarthritis; senescence-associated ophthalmic diseases and disorders; and senescence-associated dermatological diseases and disorders. 1. A method of selectively eliminating senescent cells in tissues of a brain , comprising contacting senescent cells in tissues of the brain with a senolytic agent that is an MDM2 inhibitor.2. The method of claim 1 , wherein the MDM2 senolytic agent is a small molecule inhibitor.3. The method of claim 1 , wherein the MDM2 senolytic agent is a polynucleotide or oligonucleotide.4. The method of claim 3 , wherein the MDM2 senolytic agent that is a polynucleotide or oligonucleotide is an antisense oligonucleotide claim 3 , siRNA or shRNA.5. The method of claim 1 , wherein the MDM2 senolytic agent is administered to the senescent cells in tissues of the brain intermittently claim 1 , wherein the intermittent administration is a treatment cycle followed by a non-treatment interval.6. The method of claim 5 , wherein the non-treatment interval is between at least about 0.5 to 12 months.7. The method of claim 2 , wherein the MDM2 senolytic small molecule inhibitor is administered to the senescent cells in tissues of the brain intermittently claim 2 , wherein the intermittent administration is a treatment cycle followed by a non-treatment interval.8. The method of claim 7 , wherein the non-treatment interval is between at least about 0.5 to 12 months.9. The method of claim 4 , wherein the MDM2 senolytic antisense oligonucleotide is administered to the senescent cells in tissues of ...

TRUNCATED VARIANT OF THE MAMMALIAN TARGET FOR RAPAMYCIN (mTOR) PROTEIN

The invention relates to mTORbeta, a splice form of mTOR, nucleic acids encoding mTOR beta, and antibodies against mTOR beta. The invention also relates to methods of producing mTOR beta and methods of screening for an agent that modulates mTOR beta expression and/or activity. The invention further relates to a method of treating a disease associated with aberrant expression of mTOR beta by administration of an agent that alters mTOR activity and/or expression.

Leptin Compositions and Methods for Treating Progressive Cognitive Function Disorders Resulting from Accumulation of Neurofibrillary Tangles and Amyloid Beta

The present disclosure provides compositions containing a leptin product and methods of clinical therapy and diagnostic methods for progressive cognitive disorders. According to one aspect, the described invention provides a method for treating a progressive cognitive disorder. According to another aspect, the described invention provides a method for improving resilience of cognitive function in a subject in need thereof. According to another aspect, the described invention provides a method for identifying an effective therapeutic agent for treating a progressive cognitive dysfunction disease or disorder that results from at least one of accumulation of Aβ, hyperphosphorylation of tau, or accumulation of neurofibrillary tangles.

COMPOSITIONS AND METHODS FOR INHIBITING KINASE ACTIVITY

The present invention features therapeutic compositions comprising an agent that specifically binds to a PIF pocket of Aurora A kinase and an agent that specifically binds to an ATP-binding site of Aurora A kinase, and the use of the therapeutic compositions to modulate Aurora A kinase for the treatment of cancer. 1. A pharmaceutical composition comprising an effective amount of an agent that specifically binds a PIF pocket on a kinase , an effective amount of an agent that specifically binds an ATP-binding site on the kinase , and a pharmaceutically acceptable carrier.2. The pharmaceutical composition of claim 1 , wherein the agent that specifically binds a PIF pocket and/or the agent that specifically binds an ATP-binding site is an antibody or antigen binding fragment thereof claim 1 , antibody mimetic claim 1 , peptide claim 1 , polynucleotide claim 1 , or small molecule compound.3. The pharmaceutical composition of claim 1 , wherein the kinase is Aurora A kinase.410-. (canceled)12. The pharmaceutical composition of claim 1 , wherein the agent that specifically binds the PIF pocket is PS48.13. The pharmaceutical composition of claim 1 , wherein the agent that specifically binds the ATP-binding site is Danusertib1415-. (canceled)16. A method of decreasing activity of a kinase claim 1 , the method comprising contacting a kinase with an effective amount of an agent that specifically binds a PIF pocket on the kinase and an effective amount of an agent that specifically binds an ATP-binding site of the kinase claim 1 , thereby decreasing activity of the kinase.17. A method of inhibiting development of resistance to kinase inhibition in a subject claim 1 , inhibiting growth and/or proliferation of a cancer cell claim 1 , or treating cancer associated with kinase activity claim 1 , the method comprising administering to the subject an effective amount of an agent of claim 1 , thereby preventing or inhibiting development of resistance to kinase inhibition in the subject ...

METHOD

The present invention provides a method of quantifying the activity of a protein modifying enzyme in a sample, comprising: (i) grouping modified peptides from a first sample and modified peptides from a second sample into a single group according to one of the following parameters: (a) modified peptides having a modification site that is modified by the same protein modifying enzyme; or (b) modified peptides having a modification site that is part of the same modification motif; (ii) calculating enrichment of the modified peptides from the first sample compared to the modified peptides from the second sample in the group; and (iii) calculating the statistical significance of said enrichment; wherein a statistically significant enrichment is indicative of a protein modifying enzyme being activated in the first sample compared to the second sample. In some embodiments, the method further comprises identifying modified peptides in a first sample and a second sample using mass spectrometry (MS) prior to step (i). 1. A method of antifying the activity of a protein modifying enzyme in a. sample , comprising: (a) modified peptides having a modification site that is modified by the same protein modifying enzyme; or', '(b) modified peptides having a modification site that is part of the same modification motif;, '(i) grouping modified peptides from a first sample and modified peptides from a second sample into a single group according to one of the following parameters(ii) calculating enrichment of the modified peptides from the first sample compared to the modified peptides from the second sample in the group; and(iii) calculating the statistical significance of said enrichment;wherein a statistically significant enrichment is indicative of a protein modifying enzyme being activated in the first sample compared to the second sample.2. The method according to claim 1 , wherein enrichment is calculated by counting the number of modified peptides in the group which are more ...

Assay for compounds having activity against influenza

The present invention provides a method for identifying a substance capable of inhibiting at least one activity of an RNA-dependent RNA polymerase (RdRp) comprised in an influenza viral ribonucleoprotein (RNP) complex, which method comprises or consists of: (a) providing an aqueous suspension of purified influenza virus comprising said RNP complex; (b) adding to said suspension of step (a) (i) a nonionic or zwitterionic surfactant; and (ii) a reducing agent; (c) incubating the mixture obtained in step (b) in order to obtain an influenza virus lysate; (d) contacting the influenza virus lysate obtained in step (c) with a test substance under conditions that permit said test substance to interact with said RdRp comprised in said RNP complex, said RNP complex being present in said influenza virus lysate; and (e) determining whether said test substance inhibits said at least one activity of said RdRp. 114-. (canceled)15. A method for identifying a substance capable of inhibiting at least one activity of an RNA-dependent RNA polymerase (RdRp) comprised in an influenza viral ribonucleoprotein (RNP) complex , the method comprising:(a) providing an aqueous suspension of purified influenza virus comprising said RNP complex; (i) a nonionic or zwitterionic surfactant; and', '(ii) a reducing agent;, '(b) adding to said suspension of step (a)'}(c) incubating the mixture obtained in step (b) in order to obtain an influenza virus lysate;(d) contacting the influenza virus lysate obtained in step (c) with a test substance under conditions that permit said test substance to interact with said RdRp comprised in said RNP complex, said RNP complex being present in said influenza virus lysate; and(e) determining whether said test substance inhibits said at least one activity of said RdRp.16. The method of claim 15 , wherein said activity is selected from transcription claim 15 , Cap-binding claim 15 , endonuclease activity claim 15 , replication and polymerase activity.17. The method ...

METHODS OF DIAGNOSING AND TREATING B CELL ACUTE LYMPHOBLASTIC LEUKEMIA

Methods for the diagnosis and treatment of B cell Acute Lymphoblastic Leukemia (B-ALL), based in part on the detection and/or inhibition of Focal Adhesion Kinase (FAK), e.g., phosphorylated FAK (pFAK). 1. A method of treating a subject who has B cell Acute Lymphoblastic Leukemia (B-ALL) , comprising(i) identifying a subject comprising leukemic cells having a mutation in IKZF1, wherein the mutation results in haploinsufficiency or expression of a dominant negative form of Ikaros and/or in hyperactivation of FAK activity; and(ii) administering a therapeutically effective amount of an inhibitor of Focal Adhesion Kinase (FAK).2. The method of claim 1 , wherein the inhibitor of FAK is Compound C4 (chloropyramine hydrochloride); FAK Inhibitor 14; Masitinib; PF 562271 (N-methyl-N-(3-(((2-((2-oxoindolin-5-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)methyl)pyridin-2-yl)methanesulfonamide); PF 431396 (N-Methyl-N-[2-[[[2-[(2 claim 1 ,3-dihydro-2-oxo-1H-indol-5-yl)amino]-5-(trifluoromethyl)-4-pyrimidinyl]amino]methyl]phenyl]methanesulfonamide); PF 573228 (3 claim 1 ,4-Dihydro-6-[[4-[[[3-(methylsulfonyl)phenyl]methyl]amino]-5-(trifluoromethyl)-2-pyrimidinyl]amino]-2(1H)-quinolinone); PF-00562271 claim 1 , the benzenesulfonate salt of PF-562271; VS-4718; VS-6063 (PF-04554878 claim 1 , defactinib); AG82; a 7H-pyrrolo[2 claim 1 ,3-d]pyrimidine; GSK2256098; BI1853520; TAE-226; ME-TAE-226; NVP-TAE-226; FRNK; PND-1186; TAC-544; 1 claim 1 ,2 claim 1 ,4 claim 1 ,5-Benzenetetraamine terrahydrochloride; or 2-[(5-chloro-2-[[3-methyl-1-(1-methylethyl)-1H-pyrazol-5-yl]ami-no]-4-pyridinyl)aminol-N-methoxybenzamide claim 1 , or a pharmaceutically acceptable salt thereof.3. The method of claim 1 , wherein the inhibitor of FAK is an inhibitory nucleic acid selected from the group consisting of siRNA claim 1 , shRNA claim 1 , and antisense oligonucleotides; or a dominant negative FAK protein.4. The method of claim 1 , wherein the inhibitor of FAK is administered in combination with an ABL1 ...