WO2016187347A1 - Methods for diagnosing and assessing treatment for cushing's syndrome - Google Patents

Methods for diagnosing and assessing treatment for cushing's syndrome Download PDF

Info

Publication number
WO2016187347A1
WO2016187347A1 PCT/US2016/033143 US2016033143W WO2016187347A1 WO 2016187347 A1 WO2016187347 A1 WO 2016187347A1 US 2016033143 W US2016033143 W US 2016033143W WO 2016187347 A1 WO2016187347 A1 WO 2016187347A1
Authority
WO
WIPO (PCT)
Prior art keywords
sample
gra
fkbp5
subject
activity
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2016/033143
Other languages
English (en)
French (fr)
Inventor
Joseph K. Belanoff
Hazel Hunt
John Francis UNITT
Andreas G. MORAITIS
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Corcept Therapeutics Inc
Original Assignee
Corcept Therapeutics Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to EP16797242.1A priority Critical patent/EP3297728A4/en
Priority to JP2017559803A priority patent/JP6755264B2/ja
Priority to IL309712A priority patent/IL309712A/en
Priority to KR1020247001601A priority patent/KR102876869B1/ko
Priority to MX2017014857A priority patent/MX2017014857A/es
Priority to PH1/2017/502099A priority patent/PH12017502099B1/en
Priority to CA2984623A priority patent/CA2984623C/en
Priority to US15/570,710 priority patent/US10604807B2/en
Priority to KR1020177035970A priority patent/KR20180008627A/ko
Priority to AU2016264321A priority patent/AU2016264321B2/en
Application filed by Corcept Therapeutics Inc filed Critical Corcept Therapeutics Inc
Publication of WO2016187347A1 publication Critical patent/WO2016187347A1/en
Priority to IL255612A priority patent/IL255612A/en
Anticipated expiration legal-status Critical
Priority to US15/839,666 priority patent/US11268145B2/en
Priority to US16/823,802 priority patent/US20200216905A1/en
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/565—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol
    • A61K31/567—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol substituted in position 17 alpha, e.g. mestranol, norethandrolone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
    • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/38—Drugs for disorders of the endocrine system of the suprarenal hormones
    • A61P5/46—Drugs for disorders of the endocrine system of the suprarenal hormones for decreasing, blocking or antagonising the activity of glucocorticosteroids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/533—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving isomerase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y502/00—Cis-trans-isomerases (5.2)
    • C12Y502/01—Cis-trans-Isomerases (5.2.1)
    • C12Y502/01008—Peptidylprolyl isomerase (5.2.1.8), i.e. cyclophilin
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
    • G01N33/743—Steroid hormones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/72—Assays involving receptors, cell surface antigens or cell surface determinants for hormones
    • G01N2333/723—Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/99—Isomerases (5.)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/04—Endocrine or metabolic disorders
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • Glucocorticoids are a class of steroid hormones that bind to and activate the glucocorticoid receptor (GR), which is present in almost every vertebrate cell.
  • the GR is pleiotropic and regulates a variety of important pathways in the vertebrate organism, for example, metabolism, immunity, and development. As such, detection of GR activity or regulation can be used to diagnose a variety of different vertebrate diseases, or assess a clinical or biochemical response to treatments that modulate GR activity.
  • GR activity or regulation can be used for detection of various forms of Gushing 's syndrome.
  • glucocorticoid receptor antagonists can be administered to a patient to treat a number of different diseases and conditions, and detection of a change in GR activity in response to administration of the GRA can indicate or assess a clinical or biochemical response to the treatment.
  • current methods and compositions for assessing GR activity suffer from one or more of the following insufficiencies: high cost, low sensitivity, low specificity, high false positive rate, or high false negative rate. Therefore, there remains a need for improved methods and compositions for detection of GR activity.
  • the present invention provides a method for assessing a clinical or biochemical response to a glucocorticoid receptor antagonist (GRA) in a human subject, the method comprising: a) measuring a first amount, or activity of 51 kDa FK506 binding protein (FKBP5 protein) or a first expression level of a gene encoding F BP5 protein in a first sample from the subject, wherein: i) the first sample comprises primary cells: and ii) the first sample is or was obtained before administering the GRA to the subject; b) optionally administering the GRA to the subject; c) measuring a second amount or activity of FKBP5 protein or a second expression level of a gene encoding FKBP5 protein in a second sample from the subject, wherein: i) the second sample comprises primary cells; and ii) the second sample is or was obtained after administering the GRA to the subject; and d) comparing the first and second amounts, activities, or expression levels, where
  • the present invention provides a method for assessing a clinical or biochemical response to a GRA in a human subject, the method comprising: a) measuring a first amount, or activity of 51 kDa FK506 binding protein (FKBP5 protein) or a first expression level of a gene encoding FKBP5 protein in a first sample from the subject, wherein: i) the first sample comprises primary cells; and ii) the first sample is or was obtained before administering the GRA to the subject; b) optionally administering the GRA to the subject; c) measuring a second amount or activity of FKBP5 protein or a second expression level of a gene encoding FKBP5 protein in a second sample from the subject, wherein: i) the second sample comprises primary ceils; and ii) the second sample is or was obtained after administering the GRA to the subject; d) comparing the first and second amounts, activities, or expression levels to obtain an FKBP5 difference value; e) comparing
  • FKBP5 protein 51
  • the threshold difference value is a threshold reduction value and a presence of a reduction in FKBP5 amount or activity between the first and second sample that is greater than a threshold reduction value indicates a clinical or biochemical response to the GRA.
  • the threshold difference value is a threshold reduction value and a presence of a reduction in FKBP5 amount or activity between the first and second sample that is similar to (e.g. , within about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, or 70% of) a threshold reduction value indicates a clinical or biochemical response to Z die GRA.
  • the at least 20 or 30 or 50 test individuals are subjects known to have or suspected of having Gushing 's syndrome.
  • the threshold difference value is dete mined by subtracting a post-GRA administration FKBP5 amount, activity, or expression from a pre-GRA administration FKBP5 amount, activity, or expression in a cohort of 20 or 30 or 50 test individuals known to have or suspected of having Gushing 's syndrome that is controlled or well-controlled by GRA administration.
  • the subject is administered a GRA in multiple doses and the first sample is obtained prior to administering the multiple doses of GRA and the second sample is obtained after administering the multiple doses of GRA to the subject.
  • the measuring of a) and/or c) comprises quantitating an amount of mRNA encoding FKBP5 protein in the sample.
  • the measuring of a) and/or c) comprises quantitating the amount of FKBP5 protein in the sample.
  • the measuring comprises quantitating the amount of FKBP5 protein activity in the sample.
  • the quantitating the amount of FKBP5 protein activity in the sample comprises measuring FKBP5 protein peptidyl-prolyl-cis-trans isomerase activity in the sample. In some embodiments, the quantitating the amount of FKBP5 protein activity in the sample comprises measuring die amount of FKBP5 protein bound to glucocorticoid receptor (GR) in the sample.
  • GR glucocorticoid receptor
  • the administering the GRA to the subject comprises administering mifepristone to the subject. In some embodiments, the administering the GRA to the subject comprises administering a GRA that is not mifepristone to the subject. In some embodiments, die administering the GRA to the subject comprises administering a heteroaiyl-ketone GRA.
  • the first or second samples comprise whole blood, or a fraction thereof. In some embodiments, the first and second samples comprise whole blood, or a fraction thereof. In some embodiments, the first or second samples comprise nasal epithelial scraping samples.
  • the patient is in need of administration of the glucocorticoid receptor antagonist (GRA). In some embodiments, the patient has elevated levels of Cortisol.
  • GAA glucocorticoid receptor antagonist
  • the patient has cancer and the first and second samples comprise tumor cells.
  • the first or second samples comprise whole blood, or a fraction thereof.
  • the first and second samples comprise whole blood, or a fraction thereof.
  • the method comprises administering an increased amount of GRA to the subject in the absence of a detected reduction in the amount or activity of FKBP5 protein or a detected reduction in the expression level of the gene encoding FKBP5 protein in the second sample.
  • the present invention provides a method for diagnosing Gushing 's syndrome in a human subject, the method comprising: a) measuring an amount, or activity of 51 kDa FK506 binding protein (FKBP5 protein) or an expression level of a gene encoding FKBP5 protein in a sample obtained or provided from the subject, wherein the sample comprises primary cells; and b) identifying the subject as likely to be suffering from
  • the control comprises a value derived from at least 100 or at least 200 test individuals.
  • the test individuals are subjects that are known to not exhibit Cushing's syndrome.
  • the test individuals are subjects that are known to have normal Cortisol levels or are known to lack hypercortisolemia.
  • the control comprises a value derived from at least 20 or 30 or 50 test individuals.
  • the test individuals are subjects diagnosed with Cushing's syndrome and undergoing therapy with a GRA.
  • the test individuals are subjects diagnosed with Cushing's syndrome, undergoing therapy with a GRA, wherein at least one symptom of the Cushing's syndrome is mitigated or eliminated by the GRA therapy.
  • the present invention provides a method for assessing a clinical or biochemical response to a GRA in a human subject, the method comprising: a) measuring a amount, or activity of 51 kDa FK506 binding protein (FKBP5 protein) or an expression level of a gene encoding FKBP5 protein in a sample from the subject, wherein: i) the sample comprises primary ceils; and li) the sample is or was obtained after administering the GRA to the subject; d) comparing the amount, activities, or expression level of FKBP5 to a control value derived from a cohort of at least 100 or at least 200 test individuals; and f) identifying tlie subject as having or not having the clinical or biochemical response to the GRA based on a comparison of the FKBP5 amount, activity or expression level to tlie control value.
  • FKBP5 protein 51 kDa FK506 binding protein
  • the at least 100 or at least 200 test individuals are normal subjects that are not otherwise in need of a GRA. In some cases, the at least 100 or at least 200 test individuals do not have Cushing's syndrome. In some cases, the at least 100 or at least 200 test individuals do not have elevated levels of Cortisol. In some cases, the at least 100 or at least 200 test individuals do not have, or do not exhibit symptoms of hypercortisolemia.
  • the measuring comprises quantitating an amount of mRNA encoding FKBP5 protem in the primary cells of the sample. In some embodiments, the measuring comprises quantitating the amount of FKBP5 protein in the primary cells of the sample.
  • the measuring comprises quantitating the amount of FKBP5 protein activity in the primary cells of the sample.
  • the quantitating the amount of FKBP5 protein activity in the primary cells of the sample comprises quantitating FKBP5 protein peptidyl-proiyl-cis-trans isomerase activity' in the primary cells of the sample.
  • the quantitating the amount of FKBP5 protein activity in the primary cells of the sample comprises quantitating the amount of FKBP5 protein bound to G in the primary cells of the sample.
  • the sample obtained from the subject comprises whole blood, or a fraction thereof.
  • the subject has undergone transsphenoidal surgery before the sample is obtained from the subject.
  • the sample is obtained from, the subject less than eleven days after the subject is treated with transsphenoidal surgery. In some embodiments, the sample is obtained from die subject less than two, four, or six weeks after the subject is treated with transsphenoidal surgery. In some embodiments, the method comprises administering a treatment for Cushing's syndrome when the amount or activity of FKBP5 protein or the expression level of the gene encoding FKBP5 protein is high relative to a control. In some cases, the administering the treatment for Cushing's syndrome comprises administering to the subject a glucocorticoid receptor antagonist (GRA)
  • GAA glucocorticoid receptor antagonist
  • the present invention provides a method for assessing a clinical or biochemical response in a human subject to administering to the subject a medical or surgical therapy for treatment of hypercortisolemia, the method comprising: a) measuring a first amount, or activity of 51 kDa FK506 binding protein (FKBP5 protein) or a first expression level of a gene encoding FKBP5 protein in a first sample from the subject, wherein: i) the first sample comprises primary cells; and ii) the first sample is or was obtained before administering the medical or surgical therapy for treatment of hypercortisolemia to the subject: b) optionally administering the medical or surgical therapy for treatment of hypercortisolemia to the subject; c) measuring a second amount or activity of FKBP5 protein or a second expression level of a gene encoding FKBP5 protein in a second sample from the subject, wherein: i) the second sample comprises primary cells; and ii) the second sample is or was obtained
  • a reduction in the amount or activity of FKBP5 protein or a reduction in the expression level of the gene encoding FKBP5 protein in the second sample can indicate that the medical or surgical therapy is successful in treating the liypercortisolism
  • the present invention provides a method for assessing a clinical or biochemical response to administering to the subject a medical or surgical therapy for treatment of hypercortisolemia, the method comprising: a) measuring a first amount, or activity of 51 kDa FK506 binding protein (FKBP5 protein) or a first expression level of a gene encoding FKBP5 protein in a first sample from, the subject, wherein: i) the first sample comprises primary cells; and ii) the first sample is or was obtained before administering the GRA to the subject; b) optionally administering the medical or surgical therapy for treatment of hypercortisolemia to the subject; c) measuring a second amount or activity of FKBP5 protein or a second expression level of a gene encoding FKBP5 protein in a second sample from die subject, wherein: i) the second sample comprises primary cells; and ii) the second sample is or was obtained after administering the GRA to the subject; d) comparing the first amount, or activity of 51 kD
  • the threshold difference value is a threshold reduction value and a presence of a reduction in FKBP5 amount or activity between the first and second sample that is greater than, or similar to, a threshold reduction value indicates a clinical or biochemical response to the GRA.
  • the at least 20 or 30 or 50 test individuals are subjects known to have or suspected of having Cushing's syndrome.
  • hypercortisolemia is selected from the group consisting of: inhibition of steroidogenesis, administration of an ACTH modulator, GRA administration, : transsphenoidal surgery, repeat transsphenoidal surgery, unilateral adrenalectomy, bilateral adrenalectomy, radiotherapy, resection of a non-pituitary ACTH-secreting tumor, treatment with a peptide receptor radionuclide therapy, and combinations thereof.
  • the inhibition of steroidogenesis comprises administration of ketoconazole, levoketoconazole, metyrapone, LCI699, mitotane, aminoglutethimide, etomidate, or a combination thereof
  • the administration of an ACTH modulator comprises administration of a dopamine agonist, somatostatin, a somatostatin analog, retinoic acid, R-roscovitine, or a combination thereof.
  • the dopamine agonist is selected from the group consisting of bromocriptine and cabergoline.
  • the metliod comprises administering an additional medical or surgical therapy for treatment of hypercortisolemia in an absence of a detected reduction in the amount or activity of FKBP5 protein or reduction in the expression level of the gene encoding FKBP5 protein in the second sample.
  • the measuring of a) and/or c) comprises quantitating an amount of mRNA encoding FKBP5 protein in the sample. In some embodiments, the measuring of a) and/or c) comprises quantitating the amount of FKBP5 protein in the sample. In some embodiments the measuring comprises quantitating the amount of FKBP5 protein activity in the sample.
  • the quantitating the amount of FKBP5 protein activity in the sample comprises measuring FKBP5 protein peptidyl-prolyl-cis-trans isomerase activity in the sample. In some embodiments, the quantitating the amount of FKBP5 protein activity in the sample comprises measuring the amount of FKBP5 protein bound to glucocorticoid receptor (GR) in the sample.
  • GR glucocorticoid receptor
  • the GRA administration comprises administering mifepristone to the subject. In some embodiments, the GRA administration comprises administering a GRA that is not mifepristone to the subject. In some embodiments, GRA administration comprises administering a heteroaryl-ketone GRA.
  • the first or second samples comprise whole blood, or a fraction thereof. In some embodiments, the first and second samples comprise whole blood, or a fraction thereof. In some embodiments, the first or second samples comprise nasal epithelial scraping samples.
  • the patient is in need of a medical or surgical therapy for treatment of hypercortisolemia. In some embodiments, the patient has elevated levels of Cortisol.
  • FIG. 1 depicts a study scheme for examining the expression level of the gene
  • FIG. 2 depicts the fold difference in GAPDH normalized FKBP5 expression levels in samples taken before and after administration of the indicated GR agonist (prednisone), or before and after administration of the indicated GR agonist in combination with the indicated GR antagonist (mifepristone or CORT125134).
  • GR glucocorticoid receptor'
  • NRC1 nuclear receptor subfamily 3, group C, member 1
  • the GR is also referred to as the Cortisol receptor.
  • the term includes isoforms of GR, recombinant GR and mutated GR.
  • GABA Glucocorticoid receptor antagonist
  • glucocorticoid receptor GR
  • Cortisol cortisol analogs, synthetic or natural
  • a "specific glucocorticoid receptor antagonist” refers to any composition or compound which inhibits any biological response associated with the binding of a GR to an agonist. By “specific,” the drug preferentially binds to the GR rather than other nuclear receptors, such as mineralocorticoid receptor (MR), androgen receptor ( AR), or progesterone receptor (PR).
  • MR mineralocorticoid receptor
  • AR androgen receptor
  • PR progesterone receptor
  • the specific glucocorticoid receptor antagonist binds GR with an affinity that is 10-fold greater ( 1/10 th the 3 ⁇ 4 value) than its affinity to the MR, AR, or PR, both the MR and PR, both the MR and AR, both the AR and PR, or to the MR, AR, and PR.
  • the specific glucocorticoid receptor antagonist binds GR with an affinity that is 100-fold greater ( 1/100 th the d value) than its affinity to the MR, AR, or PR, both the MR and PR, both the MR and AR, both the AR and PR, or to the MR, AR, and PR.
  • "Human subject" refers to a human primate.
  • the human subject can be a person having, or suspected of having, Gushing 's syndrome or a disease or condition that can be treated with a GRA.
  • a patient in need of administration of a glucocorticoid receptor antagonist (GRA) can be a person having or, or suspected of having, Gushing' s syndrome or a disease or condition that can be treated with a GRA.
  • GAA glucocorticoid receptor antagonist
  • Exemplary diseases or conditions that can be treated with a GRA include, but are not limited to cancer, breast cancer, triple negative breast cancer, prostate cancer, metastatic prostate cancer, ovarian cancer, Cushing's syndrome, or Cushing's disease.
  • the human subject can be a person that has been previously treated with transsphenoidal surgery (e.g.
  • the subject may have undergone transsphenoidal surgery to treat Cushing ' s disease.
  • the human subject previously treated with transsphenoidal surgery can be a subject that has undergone transsphenoidal surgery less than, or less than about, 20 days, 19 days, 18 days, 17 days, 16 days, 15 days, 14 days, 13 days, 12 days, 1 1 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day ago.
  • Assessing a clinical or biochemical response to a glucocorticoid receptor antagonist refers to detecting or quantifying a response to an administered GRA.
  • the clinical response can be an indication that the GRA is likely to be successful in treating a disease or condition, or successful in mitigating or ameliorating one or more symptoms of a disease or condition.
  • the biochemical response can be an indication that the GRA is at a dose that is sufficient to alter, or significantly alter, the physiology of the subject to which the GRA is administered.
  • the biochemical response can be an indication that the GRA is likely to be successful in treating a disease or condition, or successful in mitigating or ameliorating one or more symptoms of a disease or condition.
  • the disease or condition can be, e.g.
  • hypercortisolemia or Cushing's syndrome The clinical or biochemical response can be assessed by detecting a change in G activity or regulation caused by or correlated with administration of a GRA. For example, a change in the amount or activity of FKBP5 protein, or the expression level of a gene encoding FKBP5 protein, in response to administration of a GRA can be detected to assess a clinical response or a biochemical response to a GRA.
  • "Assessing a clinical or biochemical response to administering a medical or surgical therapy for treatment of hypercortisolemia" and the like, refers to detecting or quantitating a response to the administered therapy.
  • the clinical response can be an indication that the therapy is likely to be successful in treating the hypercortisolemia, or successful in mitigating or ameliorating one or more symptoms of the hypercortisolemia.
  • the biochemical response can be an indication that the administered therapy has altered, or significantly altered, the physiology of the subject to which the therapy is administered.
  • the biochemical response can be an indication that the therapy is likely to be successful in treating the
  • hypercortisolemia or successful in mitigating or ameliorating one or more symptoms of the hypercortisolemia.
  • Measuring an amount or activity of FKBP5 protein refers to measuring the amount of FKBP5 protein or measuring the amo unt of an acti v ity of the FKBP5 protein in a sample.
  • the activity can be cis-trans prolyl isomerase activity, FK506 or rapamycin binding activity, GR binding activity, or chaperone activity (e.g., steroid hormone chaperone activity).
  • Measuring an expression level of a gene encoding FKBP5 protein generally refers to measurement of the amount of mRNA encoding FKBP5 protein in a sample, or measuring the production of mRNA encoding FKBP5 protein in the sample.
  • Methods for measuring mRNA or mRNA production include, but are not limited to, RT-PCR, digital RT-PCR, RNA- seq (e.g., Methods Mol Biol. 2014; 1158:71-91), and microarray analysis.
  • sample refers to a biological sample obtained from any tissue or organ of a human subject.
  • the sample can be any cell or tissue sample from a human subject. Samples can be subject to various treatment, storage or processing procedures before being analyzed according to the methods described herein. Generally, the terms “sample” or “samples” are not intended to be limited by their source, origin, manner of procurement, treatment, processing, storage or analysis, or any modification.
  • the biological sample can contain primary' cells originating from a human subject.
  • the sample can contain an FKBP5 polypeptide or portion thereof, a nucleic acid encoding an FKBP5 polypeptide or portion thereof, an amplification or reverse transcription product of a nucleic acid encoding an FKBP5 polypeptide or portion thereof, or combination of any two or more of the foregoing polypeptides, nucleic acids, amplification products, reverse transcription products, or portions thereof.
  • “Whole blood” refers to blood collected from a human subject that is not subject to serum or plasma separation.
  • a fraction thereof 1 in the context of such whole blood refers to any fraction of whole blood, such as plasma, serum, a leukocyte fraction, a red blood cell fraction, a sample of peripheral blood mononuclear cells, and the like.
  • Control in the context of amount or activity of FKBP5 protein, expression levels of the FKBP5 gene, or a fold-change in FKBP5 amount, activity, or expression level, can refer to a level that is typically found in a subject under various clinical conditions.
  • the control value can be an amount typically found in a Gushing' s patient.
  • the control value can be an amount typically found in a healthy (e.g. , non-Cushing's) patient.
  • the control value can be a fold-change typically observed when a patient or cells of a patient are administered a GRA (e.g.
  • a control value can be a fold-change that is typically observed when a patient or cells of a patient are administered a GRA (e.g. , at a typical dose), wherein the patient or cells of the patient that do not exhibit a clinical response to the GRA or do not exhibit a biochemical response to the GRA.
  • control value is a normalized control value.
  • the normalized control value can be normalized against a house keeping gene (e.g. , GAPDH) or protein, or normalized against total protein or RNA (e.g. , mRNA) levels as described herein.
  • control value is an absolute value, or an absolute value per sample volume, or per number of cells in a sample.
  • the 51 -kDa FK506 binding protein (FKBP51 or FKBP5) is part of the
  • immunophilin family a superfamily of highly conserved proteins first characterized by their ability to bind to immunosuppressant drugs (Barik, 2006; Baughman et al., 1995).
  • FK506 binding immunophilins are also protein chaperones, and have the related but apparently separate ability to isomerize proline residues (PPIase activity) (Bank, 2006).
  • the FKBP5 protein is an hsp90 co-chaperone that interacts with steroid hormone receptors, including the glucocorticoid receptor (GR), the progesterone receptor, and the androgen receptor (O'Leary, 2013), It is one of the chaperones that maintains un-liganded GR in the cytoplasm, thus reducing the affinity of GR for Cortisol and reducing translocation of GR to the nucleus.
  • Proline cis-trans isomerization is important for proper protein folding, but deletion of the -terminal PPIase domain had little effect on the efficacy of the FKBP5 protein as a chaperone. It appears that the binding activity of the FKBP5 protein may be more important than its PPIase enzymatic activity in terms of GR signaling (O'Leary, 2013).
  • FKBP5 gene expression is induced by glucocorticoids (including Cortisol) as part of an intracellular ultrashort negative feedback loop for GR activity (Vermeer, 2003). This induction is mediated by GR (Vermeer, 2003; Caldwell 2010). Chronic administration of corticosterone to mice, also results in elevated FKBP5 gene expression (Lee, 2010; Ewald, 2014).
  • glucocorticoids including Cortisol
  • FKBP5 Overexpression of FKBP5 in vitro reduces glucocorticoid binding affinity and nuclear translocation of GR.
  • Naturally occurring over-expression of FKBP5 causes GR resistance in New World monkeys (Scammell, 2001), which is accompanied by increased Cortisol levels.
  • a role for FKBP5 in bone destruction, the development of osteoporosis in rheumatoid arthritis, and in glucocorticoid induced osteoporosis has been suggested (Kimura, 2013).
  • Overexpression of F BP5 in vivo in a rTg45 10 tau transgenic mouse model resulted in an increase in the level of phosphorylated tau and was associated with neuronal loss (Blair, 2013).
  • overexpression of FKBP5 has been associated with Alzheimer's disease (Blair, 2013).
  • FKBP5 polymorphisms of FKBP5 have been linked with several diseases, including depression (Binder 2004), post-traumatic stress disorder (Binder 2008), mood disorders (Binder 2009) and bipolar disorder (Willour, 2009). FKBP5 polymorphisms have also been linked to the response to anti -depressant treatment (Binder, 2004).
  • FKBP5 expression in adipose tissue is correlated with markers of insulin resistance.
  • SNPs in the FKBP5 region were associated with type 2 diabetes and diabetes-related traits.
  • GAA glucocorticoid receptor antagonist
  • the cells, tissue, or organ affected by the disease or condition can be, or can become, resistant to, or unaffected by, the administration of one or more GRAs.
  • certain breast cancer cells e.g. , triple negative breast cancer cells
  • GRA treatment e.g., in combination with one or more chemotherapeutics
  • some diseases or conditions caused by or associated with elevated levels of Cortisol exhibit variable treatment efficacy by administration of one or more GRAs.
  • identification of a clinical or biochemical response to the GRA can indicate that the GRA treatment should be continued.
  • a lack of a clinical or biochemical response can indicate that a different treatment (e.g., administration of a different GRA or administration of the same GRA at a higher dose) is indicated.
  • a different treatment e.g., administration of a different GRA or administration of the same GRA at a higher dose
  • Such responsiveness can vary from subject to subject, cell to cell, tissue to tissue, or during the course of a subject's disease progression.
  • glucocorticoid receptor antagonist GAA
  • the methods for detecting or assessing a clinical or biochemical response to a GRA described herein involve detection of: (i) the activity or amount of FKBP5 protein; or (ii) the expression level of a gene encoding the FKBP5 protein (e.g. , the absolute or relative amount of FKBP5 mRN A), in one or more provided samples from a human subject.
  • the methods described herein involve detection of, or detection of a change in,: (i) the activity or amount of FKBP5 protein; or (ii) the expression level of a gene encoding the FKBP5 protein (e.g., the absolute or relative amount of FKBP5 mRNA), in a subject.
  • a change can be detected by determining the activity or amount of FKBP5 protein, or the expression level of a gene encoding FKBP5, in a first sample from a subject and comparing the results to the results from a second sample from the subject.
  • the first sample can be provided or obtained from the subject prior to one or more steps of administering a GRA
  • the second sample can be provided or obtained from the subject after one or more steps of administering a GRA.
  • Cushing's syndrome is a condition caused by the excessive production of the glucocorticoid Cortisol by the adrenal cortex.
  • the condition is often due to the presence of a tumor or hyperplasia that exhibits unregulated secretion of adrenocorticotropic hormone (ACTH).
  • the unregulated secretion of ACTH in turn induces the adrenal glands to secrete excess Cortisol.
  • Cortisol generally participates in a negative feedback loop, in which high levels of Cortisol suppress secretion of both ACTH and Cortisol.
  • this negative regulation is not effective or absent, resulting in chronic hypercortisolemia.
  • Cushing's syndrome can be diagnosed in a variety of ways.
  • One method of diagnosis is known as the dexamethasone suppression test (DST).
  • DST dexamethasone suppression test
  • tins test the glucocorticoid receptor agonist dexamethasone is administered to a patient and Cortisol levels are measured after administration.
  • the agonist can be administered in a low dose (e.g., 1 -2 mg) to measure effects of a low dose on Cortisol levels.
  • a high dose is also administered (e.g., 8 mg) to measure the effects of a high dose on Cortisol levels.
  • ACTH levels can be determined prior to administration of dexamethasone for additional information.
  • the presence of a low ACTH level and a Cortisol level that is not suppressed by high or low dose dexamethasone indicates primary hypercortisolemia caused by, e.g., Cortisol secreting adenoma tumors in the adrenal cortex. This type of Cushing's syndrome is not typically subject to ACTH or Cortisol regulation.
  • the Cortisol cut off for the DST has been a moving target. Currently the endocrine society recommends a cut off of 1.8 ⁇ g/dl and, the AACE guidelines recently decreased the cut off to 3 ⁇ g/dl (pre viously it w as higher at 5 ⁇ g/dl). Endocrine societies in other countries use different cut offs.
  • the DST has high sensitivity but low specificity, therefore current guidelines for the diagnosis of Cushing's syndrome (CS) require at least 2 confirmatory tests (e.g. , a DST and confirmation by an alternative diagnostic method).
  • the false positive rate of the 1 mg DST is high in certain populations (severe insulin resistance, fatty liver disease, PCOS etc., see "The Diagnosis of Cushing's Syndrome: An Endocrine Society Clinical Practice Guideline (2008), published as J. Clin. Endocrin. & Metab., May 2008, 93(5): 1526-40.). Also medications that affect the metabolism of dexamethasone can influence the test (see guidelines from the Endocrine Society).
  • DST dexamethasone suppression test
  • UFC urine free Cortisol
  • DST Another alternative to the DST is based on midnight salivary Cortisol levels. This test also is less sensitive in milder forms of CS. It can be useful to diagnose early relapse of CD after transsphenoidal surgery. However, it has a lot of false positives and false negatives. It depends on whether the patient is able to perform the test properly, and inappropriate sample collection is frequently a confounding factor,
  • CRH can be useful in the differential diagnosis of Cushing's syndrome, because most patients with Cushing's disease respond to CRH, while those with other types of Cushing's syndrome generally do not.
  • these tests, and in particular the dexamethasone suppression test are widely used to diagnose Gushing syndrome and/or distinguish between Cushing's disease and oilier types of Cushing's syndrome, they are not definitive, and can yield an undesirable number of false positives and false negatives.
  • the present inventors have developed an improved method for diagnosing Cushing's syndrome.
  • the method can include detecting: (i) the activity or amount of FKBP5 protein; or (ii) the expression level of a gene encoding the FKBP5 protein, in one or more samples from a human subject.
  • Described herein is a method for assessing a clinical or a biochemical response to a glucocorticoid receptor antagonist (GRA) in a human subject.
  • the method can be useful, e.g. , for confirming the efficacy of a treatment modality, monitoring treatment outcomes, or guiding treatment decisions. For example, if a patient presents with a disease or condition known to be, or suspected of being, treatable with a GRA, then a GRA can be administered, and the clinical response to the GRA can be assessed using one or more of the methods described herein.
  • a positive indication of a clinical or biochemical response, or an indication of a strong clinical or biochemical response, to the GRA can then predict a positi ve clinical outcome, an increased likelihood of a positive clinical outcome, or suggest continuation of GRA adm inistration.
  • a negative indication of a clinical or biochemical response, or a lack of a strong clinical or biochemical response can predict a negative clinical outcome, an increased likelihood of a negative clinical outcome, or suggest administration of an increased dose of the GRA or administration of an alternative GRA.
  • a negative indication of a clinical response or a biochemical response, or a lack of a strong clinical or biochemical response can suggest a need for additional or alternative treatments.
  • the method for assessing a clinical or biochemical response to a glucocorticoid receptor antagonist (GRA) in a human subject can include: a) measuring a first amount, or activity of FKBP5 protein or a first expression level of a gene encoding FKBP5 protein in a first sample from the subject, wherein i) the first sample comprises primary cells; and ii) the first sample is obtained before administering the GRA to the subject; b) optionally administering the GRA to the subject; c) measuring a second amount or activity of FKBP5 protein or a second expression level of a gene encoding FKBP5 protein in a second sample from the subject, wherein: i) the second sample comprises primary cells: and ⁇ ) the second sample is obtained after administering the GRA to the subject; and d) comparing the first and second amounts, activities, or expression levels, wherein a reduction in the amount or activity of FKBP5 protein or a reduction in the expression level of the gene encoding F BP5
  • the subject has received one or more doses of a GRA prior to the obtaining of a first sample.
  • the subject may be undergoing GRA therapy, or may have previously undergone GRA therapy.
  • the pre-GRA administration sample may be obtained a suitable period of time after a GRA administration to allow for measurement of baseline FKBP5 le vels that are unaffected by the presence of a GRA.
  • the first sample which is obtained prior to administering the GRA to the subject, is obtained at least 4, 5, 6, 7, 8, 9, 10, 12, 16, 1 8, or 24 hours after a previous GRA administration, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days after a previous GRA
  • a suitable period of time between a previous administration of a GRA and obtaining a first sample can be determined based on the pharmacokinetics of a previously administered GRA. In some cases, a suitable period of time is a length of time sufficient for the GRA to be completely removed from, the subject. In some cases, a suitable period of time is a length of time sufficient for the GRA to have no or minimal effect on FKBP5 levels (e.g. , amount or activity of FKBP5 protein or mRNA levels).
  • the subject is administered multiple doses of a GRA between the obtaining of the first sample and the obtaining of the second sample.
  • the subject is administered, or is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, or more doses between the obtaining of the first sample and the obtaining of the second sample.
  • the subject is administered a GRA, and the second sample is obtained after a period of time suitable to allow for downreguiation of FKBP5.
  • the second sample can be obtained at least 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours after GRA administration.
  • the second sample can be obtained between doses of a multiple GRA dosage schedule.
  • the second sample is obtained immediately prior, immediately after, or during one or more GRA administrations. In any case, the second sample is obtained after at least one dose of GRA has been administered to the subject.
  • a method for assessing a clinical or a biochemical response in a human subject to administering to the subject a medical or surgical therapy for treatment of hypercortisolemia can be useful, e.g. , for confirming the efficacy of a treatment modality, monitoring treatment outcomes, or guiding treatment decisions.
  • a medical or surgical therapy can be administered, and the clinical response to the medical or surgical therapy can be assessed using one or more of the methods described herein, in some cases, a positive indication of a clinical or biochemical response, or an indication of a strong clinical or biochemical response, to the medical or surgical therapy can then predict a positive clinical outcome, an increased likelihood of a positive clinical outcome, or suggest continuation of the medical or surgical therapy.
  • a negative indication of a clinical or biochemical response, or a lack of a strong clinical or biochemical response can predict a negative clinical outcome, an increased likelihood of a negative clinical outcome, or suggest administration of an increased dose or additional doses of a medical therapy, or administration of an alternative medical or surgical therapy.
  • a negative indication of a clinical response or a biochemical response, or a lack of a strong clinical or biochemical response can suggest a need for additional or alternative treatments.
  • the method for assessing a clinical or biochemical response to a medical or surgical therapy for hypercortisolemia in a human subject can include: a) measuring a first amount, or activity of FKBP5 protein or a first expression level of a gene encoding F BP5 protein in a first sample from the subject, wherein i) the first sample comprises primary cells: and ii) the first sample is obtained before administering the medical or surgical therapy to the subject; b) optionally administering the medical or surgical therapy to the subject; c) measuring a second amount or activity of F BP5 protein or a second expression level of a gene encoding FKBP5 protein in a second sample from the subject, wherein: i) the second sample comprises primary cells; and ii) the second sample is obtained after administering the medical or surgical therapy to the subject; and d) comparing the first and second amounts, activities, or expression levels, wherein a reduction in the amount or activity of FKBP5 protein or a reduction in the expression level of the gene
  • the medical or surgical therapy includes administering an inhibitor of steroidogenesis.
  • inhibitors of steroidogenesis include, but are not limited to, aminogiutethimide, cholesterol sulfate, ketoconazoie, levoketoconazole, metyrapone, LCI699, mitotane, and eiomidate.
  • the medical or surgical therapy includes administering an ACTH modulator.
  • ACTH modulators include, but are not limited to, dopamine agonists, retinoic acid, R-roscovitine, somatostatin, and somatostatin analogues.
  • Exemplary dopamine agonists include, but are not limited to, bromocriptine, and cabergoline.
  • Exemplar ⁇ ' somatostatin analogues include, but are not limited to, pasireotide, octreotide, and Ianreotide.
  • the medical or surgical therapy is or includes GRA administration.
  • the medical or surgical therapy is or includes transsphenoidal surgery, or repeat transsphenoidal surgery.
  • the medical or surgical therapy is or includes adrenalectomy, unilateral adrenalectomy, or bilateral adrenalectomy.
  • the medical or surgical therapy is or includes radiotherapy.
  • the medical or surgical therapy is or includes resection of a non-pituitary ACTH-secreting tumor.
  • the medical or surgical therapy is or includes treatment with a peptide receptor radionuclide therapy (e.g., Y-90 or Lu-177 labeled octreotide) .
  • the medical or surgical therapy is or includes a combination of any two or more of the foregoing medical or surgical therapies.
  • the combination is a combination of two or more medical therapies.
  • the combination is a combination of two or more surgical therapies.
  • the combination is a combination of a medical therapy and a surgical therapy.
  • the subject has received one or more doses of a medical therapy for hypercortisoiemia prior to the obtaining of a first sample.
  • a medical therapy for hypercortisoiemia for hypercortisoiemia prior to the obtaining of a first sample.
  • the subject may be undergoing GRA therapy, or may have previously undergone GRA therapy.
  • the pre-medical therapy administration sample may be obtained a suitable period of time after a medical therapy administration to allow for measurement of baseline FKBP5 levels that are unaffected by the presence of the medical therapy.
  • the first sample which is obtained prior to administering the medical therapy to the subject, is obtained at least 4, 5, 6, 7, 8, 9, 10, 12, 16, 18, or 24 hours after a previous medical therapy administration, or at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 1 8, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days after a previous medical therapy administration.
  • a suitable period of time between a previous administration of a medical therapy and obtaining a fi rst sample can be determined based on the pharmacokinetics of a previously administered medical therapy.
  • a suitable period of time is a length of time sufficient for the medical therapy to be completely removed from the subject.
  • a suitable period of time is a length of time sufficient for the medical therapy to have no or minimal effect on FKBP5 levels (e.g. , amount or activity of FKBP5 protein or mRNA levels).
  • the subject is administered multiple doses of a medical therapy between the obtaining of the first sample and the obtaining of the second sample.
  • the subject is administered, or is administered at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or more doses between the obtaining of the first sample and the obtaining of the second sample.
  • the subject is administered a medical therapy
  • the second sample is obtained after a period of time suitable to allow for downregulation of FKBP5 expression, amount, or activity.
  • the second sample can be obtained at least 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours after administration of the medical therapy.
  • the second sample can be obtained between doses of a multiple dosage schedule.
  • the second sample is obtained immediately prior, immediately after, or during one or more administrations of a medical therapy.
  • the second sample is obtained after at least one dose of a medical therapy has been administered to the subject.
  • the sample can be obtained by any means known in the art.
  • the sample can be obtained by collecting a blood sample (e.g., a sample of whole blood or a fraction thereof).
  • the sample can be obtained by scraping epithelial cells (e.g., nasal epithelial cells) of a subject.
  • Samples include, but are not limited to samples of human cells and tissues, such as blood samples, cerebrospinal fluid samples, synovial tissue samples, synovial fluid samples, brain tissue samples, blood vessel samples, or tumor samples.
  • Samples encompass samples of healthy or pathological cells, tissues or stractures.
  • the sample can be provided by obtaining cells of a tissue or organ affected by a disease or condition mediated by G activity or signaling.
  • a type of cancer that can be treated with a GRA (e.g. , a GRA in combination with a chemotherapeutic)
  • a sample containing primary tumor cells can be obtained and assayed as described herein.
  • Such cancers include, but are not limited to, breast cancer, triple negative breast cancer, prostate cancer, metastatic prostate cancer, androgen resistant prostate cancer, and ovarian cancer.
  • the sample can be provided by obtaining cells of a subject suffering from hypercortisolemia.
  • the sample can be provided by obtaining cells of a tissue or organ affected by hypercortisolemia. In some cases, the sample can be provided by obtaining cells of a subject suffering from or suspected of being suffering from Gushing 's syndrome. In some cases, the sample can be provided by obtaining cells of a tissue or organ affected by, or suspected of being affected by, Gushing' s syndrome. [0061]
  • the sample can be extracted to obtain FKBP5 protein, or FKBP5 nucleic acid.
  • cells can be lysed, and the protein fraction obtained.
  • the lysate is further fractionated to purify a specified cellular compartment. For example, the cell lysate can be fractionated to obtain a cytosolic protein fraction.
  • the cell lysate can be fractionated to obtain a nuclear or nucleolar protein fraction.
  • the protein fraction can be assayed for FKBP5 protein levels or activity.
  • the cells can be lysed, and a nucleic acid (e.g. , mRNA) fraction obtained.
  • the nucleic acid fraction can be assayed for expression of the gene encoding FKBP5.
  • the expression can be assayed by quantitative amplification (e.g. , qPCR), or reverse transcription and subsequent quantitative amplification (e.g. , RT-qPCR).
  • the cell lysate, or protein fraction thereof can be purified using a variety of methods to obtain a fraction enriched for FKBP5 protein, or a portion thereof.
  • cells can be lysed and contacted with a chromatography medium under conditions suitable to preferentially bind contaminants or target protein. Where contaminants are preferentially bound, target protein can be collected as a flow through fraction and assayed further. Where target protein is bound, the chromatography medium can be washed and the target protein eluted.
  • cells can be lysed and contacted with a capture reagent (e.g., a capture antibody or aptamer) that specifically binds to the FKBP5 protein, or a portion thereof.
  • a capture reagent e.g., a capture antibody or aptamer
  • the capture reagent can be immobilized on a solid support. In some cases, the
  • FKBP5 protein or portion thereof can be eluted from, the capture reagent and then detected or quantified. In other cases, the FKBP5 protein or portion thereof can be detected or quantified as a capture reagent-bound form.
  • the cell lysate, or nucleic acid fraction thereof can be purified using a variety of methods to obtain a fraction enriched for a transcript of a gene encoding FKBP5.
  • cells can be lysed and nucleic acids can be precipitated or otherwise purified from the lysate.
  • the nucleic acids can be purified by contacting the sample, or a fraction thereof, with a surface immobilized oligodT moiety to preferentially bind poiyadenylated mRNA.
  • Nucleic acids can be subject to amplification, hybridization, polymerization, reverse transcription, or a combination thereof.
  • the amplification, hybridization, polymerization, or reverse transcription is target specific such that the gene encoding FKBP5, a transcript thereof, or a portion thereof is specifically amplified, hybridized, or reverse transcribed.
  • the amplification, hybridization, polymerization, or reverse transcription is not target specific such that the sample is subject to whole genome or whole transcriptome hybridization, polymerization, or reverse transcription.
  • Whole genome or other non-specific hybridization, polymerization, or reverse transcription can be performed with the use of one or more degenerate primers or probes. After non-specific hybridization, polymerization, or reverse transcription, the gene encoding FKBP5 can be detected and/or quantified.
  • the amount or activity of FKBP5 polypeptide, or a portion thereof can be measured by a variety of methods known in the art.
  • an ELISA e.g., sandwich ELISA
  • sandwich ELISA can be used to measure polypeptide levels in a sample, or a protein extract thereof, using one or more antibodies specific for the FKBP5 protein, or a portion thereof.
  • the ELISA is a sandwich ELISA, in which FKBP5 polypeptide, or a portion thereof, is immobilized by binding to an immobilized capture reagent (e.g., capture antibody or aptamer), and the immobilized polypeptide or portion thereof is detected with a detection reagent (e.g. , detection antibody or aptamer).
  • an immobilized capture reagent e.g., capture antibody or aptamer
  • activity of the FKBP5 polypeptide, or a portion thereof can be measured by contacting the sample, or a protein extract thereof, with a proline containing peptide substrate to measure the FKBP5 -mediated cis-trans prolyl isomerase activity of the sample.
  • the isomerization of the substrate can be measured using an cis-trans proline isomer sensitive enzyme, such as a protease.
  • chymotrypsin which has a high substrate specificity and catalytic efficiency (k cat /Km) against peptide substrates having a trans-proline at the P2 position and phenylalanine or tyrosine at the P 1 position, but very little or no specificity or catalytic efficiency against such peptides containing a cis-proline at P2, can be used to measure prolyl isomerase activity.
  • an N-succinyl-Ala-Leu- c s-Pro-Phe-p-nitroanilide substrate can be used in combination with chymotrypsin, which preferably cleaves the trans prolyl isomer of the substrate to assay a sample or extract thereof for FKBP5 prolyl isomerase activity.
  • the production of the trans isomer of the substrate by isomerase activity of the FKBP5 or portion thereof can be measured by detecting the digestion of the nitroanilide substrate by the chymotrypsin as described, e.g., in Fischer et al. Nature. 1989 Feb 2;337(6206):476-8.
  • activity of the FKBP5 polypeptide, or a portion thereof can be detected or quantified by measuring the amount of FKBP5 protein bound to glucocorticoid receptor (GR) in the sample.
  • GR glucocorticoid receptor
  • This can be performed, e.g. , by purifying GR or FKBP5 protein under conditions suitable to preserve binding between GR and FKBP5 protein.
  • the purification product can then be assayed for the presence, absence, or quantity of the cognate binding partner.
  • cytosolic GR can be purified and the presence of FKBP5 protein detected in the purification product.
  • FKBP5 protein can be purified and the presence of GR can be detected in the purification product.
  • this can be performed using a sandwich ELISA-type assay in which the immobilized capture reagent recognizes one member of the GR:FKBP5 protein complex and the detection reagent recognizes the other member of the GR:FKBP5 protein complex.
  • expression of a gene encoding FKBP5 can be measured by reverse transcription of FKBP5 mRNA, or a portion thereof, and quantitative amplification of tlie reverse transcription product or a portion thereof.
  • the quantitative amplification can be performed using PGR (e.g., real time PGR) or other amplification techniques known in the art.
  • the amplification can be detected by, e.g., detecting incorporation of an intercalating dye into the amplification product, degradation of a quenched fluorescence hydrolysis probe, or binding of quenched molecular beacon.
  • any measured amount or activity or expression level in a sample can be normalized to a reference.
  • an expression level can be normalized to the amount of total mRNA, or the expression level of a reference gene.
  • Suitable reference genes include, but are not limited to, GAPDH, hypoxanthine phosphoribosyltransferase 1 (HRPT1), ribosornal protein large PI, or another housekeeping gene.
  • FKBP5 protein amount can be normalized to total protein levels, or the level of a reference gene product.
  • Suitable reference gene products include, but are not limited to, actin, tubulin, COX IV, HRPT1 , GAPDH, or another housekeeping gene product. ii. Comparing F BP5 amount activity or expression
  • the amount or activity of FKBP5 or the expression level of a gene encoding FKBP5 protein after administration of a GRA is quantified from a sample obtained after administration of one or more doses of the GRA and compared to a control or threshold value rather than a pre-GRA administration value.
  • the control or threshold value is a positive control or threshold value that indicates a biochemical or clinical response, or a strong biochemical or clinical response to the GRA in the tissue, organ, or cell- type subject to the assay.
  • a quantified amount, activity, or expression level that is near, equal to, or below the positive control or threshold value can indicate a biochemical or clinical response or a strong biochemical or clinical response to the GRA.
  • a quantified amount, activity, or expression level that is above, or significantly- above, the positive control or threshold value can indicate a lack of a biochemical or clinical response, or a lack of a strong biochemical or clinical response to the GRA.
  • the quantified amount, activity, expression that indicates a biochemical or clinical response to the GRA is equal to about, or less than about, 25%, 30%, 40%, 50%, 75%, 100%, 1 10%, 125%, or 150% of the positive control or threshold value.
  • the quantified amount, activity, or expression that indicates a lack of a biochemical or clinical response to the GRA is equal to about or above about a 2-fold, 2.5- fbld, 3-fold, 4-fold, 5-fold, 7.5-fold, 10-fold, 12-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40- fold, or 50-fold multiple of the positive control or threshold value.
  • control or threshold value is a negative control or threshold value that indicates a lack of a response (e.g. , biochemical or clinical) to a GRA or a value that is typically obtained without, or prior to, administration of a GRA.
  • a quantified amount, activity, or expression level that is below the negative control or threshold value can indicate a clinical or biochemical response or a strong clinical or biochemical response to the GRA.
  • a quantified amount, activity, or expression level that is near, equal to, or above the negative control or threshold value can indicate a lack of a clinical response, a lack of biochemical response, a lack of a strong clinical response, or a lack of a strong biochemical response to the GRA.
  • the quantified amount, activity, or expression that indicates a clinical response to the GRA or a biochemical response to the GRA is equal to about or less than about 0.25%, 0.5%, 1 %, 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 25%, 30%, 40%, or 50% of the negative control or threshold value.
  • the quantified amount, activity, or expression that indicates a lack of a clinical response or a lack of a biochemical response to the GRA is at least about 75%, 100%, 1 10%, 125%, 150%, 200%, 300%, or 400% of the negative control or threshold value.
  • the amount or activity of FKBP5 protein or the expression level of a gene encoding FKBP5 after administration of a GRA is quantified from a sample obtained after administration of one or more doses of the GRA and compared to a value quantified from a sample obtained prior to administration of one or more doses of the GRA.
  • a quantified post-admin stration amount activity or expression level that is below the pre- administration value can indicate a clinical response, a biochemical response, a strong clinical response, or a strong biochemical response to the GRA.
  • a quantified post- administration amount activity or expression level that is near or equal to the pre- admini station value can indicate a lack of a clinical response, a lack of a biochemical response, a lack of a strong biochemical response, or a lack of a strong clinical response to the GRA.
  • the post-GRA administration value is greater than the pre- GRA administration value, progression of the disease or condition in the human subject is indicated.
  • the post-GRA administration value is less than the pre-GRA administration value, improvement of the disease or condition in the human subject is indicated.
  • the provided pre-GRA administration sample can be taken at any time prior to administration of a GRA, including but not limited to, immediately before GRA
  • the provided pre-administration sample is obtained from about 0 to about 12, from about 0 to 6 hours, from about 0 to about 4 hours, from about 1 to 4 hours, or from about 2 to 6 hours prior to administration of the GRA.
  • the provided post-GRA administration sample can be taken after a time suitable to allow the activity of the GRA to manifest in a change (e.g. , a detectable change) in FKBP5 amount or activity, or a change in expression of a gene encoding FKBP5.
  • the time is selected to achieve a maximum possible response (e.g. , clinical or biochemical response) to GRA administration.
  • the delay between administration of the GRA and obtaining of the post-GRA administration sample is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 15, 20, 25, 30, 40, or 50 minutes; 1 , 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 18, 20, or 22 hours; I, 2, 3, 4, 5, or 6 days; or I, 2, 3, or 4 weeks.
  • the provided post-GRA administration sample can be taken after multiple administrations of one or more GRAs.
  • one or more GRAs can be administered to a human subject in need thereof for a period of days (e.g., 1, 2, 3, 4, 5, or 6 days) or weeks ⁇ e.g.
  • the quantified post-GRA administration amount, activity, or expression that indicates a clinical or biochemical response to the GRA is less than about, 300%, 250%, 200%, 150%, 100%, 75%, 50%, 40%, 30%, 25%, 20%, 15%, 10%, 7.5%, 5%, 2.5%, 2%, 1.5%, or 1% of the pre-GRA administration value.
  • the quantified post-GRA administration amount activity or expression that indicates a lack of a clinical or biochemical response to the GRA is equal to about or above about 75%, 80%, 85%, 90%, 100%, 1 10%, 120%, 125%, 150%, or 200% of the pre-GRA administration value.
  • the human subject can have cancer, and methods described herein can be employed to assess or predict the effect of GRA therapy on the cancer cells of the subject.
  • cancer cells are obtained from a subject and assayed for FKBP5 amount or activity or the expression level of a gene encoding FKBP5.
  • a high level of FKBP5 relative to a control may suggest that the ceils express a high level of GR and GRA therapy may be indicated.
  • a low level of FKBP5 relative to a control may suggest that the cells express a low level of GR and GRA therapy is not indicated.
  • a low level of F BP5 relative to a control may suggest that the FKBP5 feedback mechanism is inoperative, and thus GRA therapy may be indicated.
  • a reduction in FKBP5 in tumor cells after administration of GRA as compared to a pre- administration value can suggest that GRA therapy is indicated.
  • FKBP5 amount, activity, or expression level can be assessed in combination with GR amount, activity, or expression level.
  • low GR amount, activity, or expression level and low FKBP5 amount, activity, or expression level can predict that GRA therapy is not beneficial for this cancer type, cell, or tumor.
  • high GR amount, activity, or expression level and high FKBP5 amount, activity, or expression level can indicate that GRA therapy can be beneficial for this cancer type, cell, or tumor.
  • high GR amount, activity, or expression level and low FKBP5 amount, activity, or expression level can indicate a defective Cortisol counter regulation mechanism and GRA therapy can be a beneficial treatment.
  • F BP5 levels can be detected in tumor cells that have been extracted from a patient and cultured in vitro.
  • a sample containing tumor ceils can be obtained, a portion of the sample assayed for FKBP5 (protein or expression), and a GRA administered to a different portion of the sample.
  • the GRA administered sample can be assayed for FKBP5 protein amount, or activity, or FKBP5 gene expression and compared to the pre-administration value.
  • F BP5 levels can be determined after transsphenoidal surgery to detect relapse or remission of Cushing's syndrome.
  • FKBP5 can be measured in samples obtained before and after transsphenoidal surgery.
  • FKBP5 can be detected in samples obtained at several time points after transsphenoidal surgery.
  • FKBP5 can be detected in one or more samples obtained within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13,14, 15, 16, 17, 18, 19, or 20 days after transsphenoidal surgery.
  • a low level of FKBP5 relative to a control can indicate that the subject is exhibiting remission of Cushing's syndrome or is not exhibiting early relapse of Cushing's syndrome.
  • a decrease in FKBP5 as compared to the value detected in a sample obtained prior to transsphenoidal surgery indicates that the subject is exhibiting remission of Cushing's syndrome or is not exhibiting early relapse of Cushing's syndrome.
  • a stable or decreasing value of FKBP5 in samples taken at multiple time points after transsphenoidal surgery indicates that the subject is exhibiting remission of Cushing's syndrome or is not exhibiting early relapse of Cushing's syndrome.
  • the method comprises administering an increased amount of GRA to the subject, or
  • the method comprises administering an increased amount of GRA to the subject, or administering an alternative GRA to the subject, or a combination thereof.
  • the amount or activity of FKBP5 or the expression level of a gene encoding FKBP5 protein after administration of a medical (e.g. , GRA) or surgical therapy for treatment of hypercortisolemia is quantified from a provided sample obtained after administration of one or more doses of a medical therapy for treatment of
  • control or threshold value is a positive control or threshold value that indicates a biochemical or clinical response, or a strong biochemical or clinical response to the medical or surgical therapy in the tissue, organ, or cell-type subject to the assay.
  • a quantified amount, activity, or expression level that is near, equal to, or below the positive control or threshold value can indicate a biochemical or clinical response or a strong biochemical or clinical response to the medical or surgical therapy for treatment of hypercortisolemia.
  • a quantified amount, activity, or expression level that is above, or significantly above, the positive control or threshold value can indicate a lack of a biochemical or clinical response, or a lack of a strong biochemical or clinical response to the medical or surgical therapy for treatment of hypercortisolemia.
  • the quantified amount, activity, expression that indicates a biochemical or clinical response to the medical or surgical therapy for treatment of hypercortisolemia is equal to, about, or less than about, 25%, 30%, 40%, 50%, 75%, 100%, 1 10%, 125%, or 150% of the positive control or threshold value.
  • the quantified amount, activity, or expression that indicates a lack of a biochemical or clinical response to the medical or surgical therapy for treatment of hypercortisolemia is equal to, about, or above about a 2-fold, 2.5-fold, 3-fold, 4-fold, 5-fold, 7.5-fold, 10-fold, 12-fold, 15- fold, 20-fold, 25-fold, 30-fold, 40-fold, or 50-fold multiple of the positive control or threshold value.
  • control or threshold value is a negative control or threshold value that indicates a lack of a response (e.g. , biochemical or clinical) to a medical or surgical therapy for treatment of hypercortisolemia or a value that is typically obtained without, or prior to, administration of a medical or surgical therapy for treatment of hypercortisolemia.
  • a quantified amount, activity, or expression level that is below the negative control or threshold value can indicate a clinical or biochemical response or a strong clinical or biochemical response to the medical or surgical therapy for treatment of hypercortisolemia.
  • a quantified amount, activity, or expression level that is near, equal to, or above the negative control or threshold value can indicate a lack of a clinical response, a lack of biochemical response, a lack of a strong clinical response, or a lack of a strong biochemical response to the medical or surgical therapy for treatment of hypercortisolemia.
  • the quantified amount, activity, or expression that indicates a clinical response to the medical or surgical therapy for treatment of hypercortisolemia or a biochemical response to the medical or surgical therapy for treatment of hypercortisolemia is equal to, about, or less than about 0.25%, 0.5%, 1%, 5%, 7.5%, 10%, 12.5%, 15%, 17.5%, 20%, 25%, 30%, 40%, or 50% of the negative control or threshold value.
  • the quantified amount, activity, or expression that indicates a lack of a clinical response or a lack of a biochemical response to the medical or surgical therapy for treatment of hypercortisolemia is at least about 75%, 100%, 110%, 125%, 150%, 200%, 300%, or 400% of the negative control or threshold value.
  • the amount or activity of FKBP5 protein or the expression le vel of a gene encoding FKBP5 after administration of a medical therapy for treatment of hypercortisolemia is quantified from a sample obtained after administration of one or more doses of the medical therapy for treatment of hypercortisolemia and compared to a value quantified from a sample obtained prior to administration of one or more doses of the medical therapy for treatment of hypercortisolemia.
  • a quantified post-administration amount activity or expression level that is below the pre -administration value can indicate a clinical response, a biochemical response, a strong clinical response, or a strong biochemical response to the medical therapy for treatment of hypercortisolemia.
  • a quantified post- administration amount activity or expression level that is near or equal to the pre- administration value can indicate a lack of a clinical response, a lack of a biochemical response, a lack of a strong biochemical response, or a lack of a strong clinical response to the medical therapy for treatment of hypercortisolemia.
  • the post- administration value is greater than the pre -administration value, progression of the hypercortisolemia condition in the human subject is indicated.
  • the post- administration value is less than the pre-administration value, improvement of the hypercortisolemia condition in the human subject is indicated.
  • the pre-administration sample can be taken at any time prior to administration of a medical or surgical therapy for treatment of hypercortisolemia, including but not limited to, immediately before therapy administration; at least about 1, 2, 3, 4, 6, 7, 8, 10, 15, 20, 25, 30, 45, 50, 60, 80, or 90 minutes before therapy administration: at least about 2, 3, 4, 5, 6, 7, 8,
  • the pre-administration sample is obtained from about 0 to about 12, from about 0 to 6 hours, from about 0 to about 4 hours, from about 1 to 4 hours, or from about 2 to 6 hours prior to administration of the therapy.
  • the post-therapy administration sample can be taken after a time suitable to allow the activity of the therapy to manifest in a change (e.g., a detectable change) in FKBP5 amoimt or activity, or a change in expression of a gene encoding FKBP5.
  • the time is selected to achieve a maximum possible response (e.g., clinical or biochemical response) to therapy administration.
  • the delay between administration of the therapy and obtaining of the post-therapy administration sample is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 20, 25, 30, 40, or 50 minutes; or 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 18, 20, 22, or 24 hours.
  • the post-therapy administration sample can be taken after multiple administrations of one or more medical or surgical therapies.
  • one or more medical therapies e.g., administration of a GRA, ACTH-modulator, inhibitor of steroidogenesis, etc.
  • one or more surgical therapies e.g.
  • transsphenoidal surgery can be administered to a human subject in need thereof and then repeated, and then a post-administration sample obtained and analyzed for FKBP5 protein amount or activity or FKBP5 gene expression to assess a clinical response to the surgical therapy or to assess a biochemical response to the surgical therapy.
  • the quantified post-administration amount, activity, or expression that indicates a clinical or biochemical response to the therapy is less than about, 75%, 50%, 40%, 30%, 25%, 20%, 15%, 10%, 7.5%, 5%, 2.5%, 2%, 1.5%, or 1% of the pre- administration value.
  • the quantified post-administration amoimt activity or expression that indicates a lack of a clinical or biochemical response to the therapy is equal to, about, or above about 80%, 85%, 90%, 100%, 110%, 120%, 125%, 150%, or 200% of the pre -administration value.
  • F BP5 levels can be determined after transsphenoidal surgery to detect relapse or remission of hypercortisolemia.
  • FKBP5 can be measured in samples obtained before and after transsphenoidal surgery. Additionally or alternatively.
  • FKBP5 can be detected in samples obtained at several time points after transsphenoidal surger -.
  • FKBP5 can be detected in one or more samples obtained within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, or 20 days after transsphenoidal surgery.
  • a low level of FKBP5 relative to a control can indicate that the subject is exhibiting remission of hypercortisolemia or is not exhibiting early relapse of hypercortisoleniia.
  • a decrease in FKBP5 as compared to the value detected in a sample obtained prior to transsphenoidal surger ' indicates that the subject is exhibiting remission of hypercortisolemia or is not exhibiting early relapse of hypercortisolemia.
  • a stable or decreasing value of FKBP5 in samples taken at multiple time points after transsphenoidal surgery indicates that the subject is exhibiting remission of hypercortisolemia or is not exhibiting early relapse of hypercortisolemia
  • the method comprises administering an increased amount of a medical therapy to the subject, administering an alternative medical therapy to the subject, administering an additional surgical therapy to the subject, or administering an alternative surgical therapy to the subject, or a combination thereof.
  • the method comprises administering an increased amount of a medical therapy to the subject, administering an alternative medical therapy to the subject, administering an additional surgical therapy to the subject, or administering an alternative surgical therapy to the subject, or a combination thereof.
  • FKBP5 amount, expression, or activity can be compared to various threshold, control, or threshold difference values to assess clinical or biochemical response as described herein.
  • a change in FKBP5 amount, expression, or activity from pre- administration e.g., pre-GRA administration or pre-medical or surgical therapy for treatment of hypercortisolemia administration
  • pre- administration e.g., pre-GRA administration or pre-medical or surgical therapy for treatment of hypercortisolemia administration
  • the threshold, control, or threshold difference value is selected to provide a sufficient sensitivity or specificity for use as a diagnostic.
  • the threshold, control, or threshold difference value can be selected to provide at least a 90% sensitivity, at least a 95% specificity, or the combination thereof.
  • the threshold, control, or threshold difference value is derived from a cohort of test individuals, wherein the cohort is of sufficient size to provide the sufficient sensitivity or specificity for use as a diagnostic.
  • the cohort size is at least 100 or at least 200 test individuals.
  • 100 or 200 healthy (e.g. , non -Gushing 's, non-hypercortisolemic, or not otherwise in need of a GRA) test individuals can be selected to provide a threshold or control amount, activity, or expression level of FKBP5.
  • a measured FKBP5 amount, activity, or expression level in a human subject that is high as compared to a threshold or control level derived from a cohort of at least 100 or at least 200 healthy test individuals can indicate the subject has Gushing' s syndrome.
  • a cohort of such healthy individuals would be expected to exhibit a small difference in FKBP5 amount, expression, or activity in response to administration of a medical or surgical therapy such as a GRA.
  • a measured reduction in FKBP5 amount, activity, or expression level from pre -administration to post-administration that is greater than (e.g. , at least 2-fold, 3-fold, 5- fold, 10-fold, 25 -fold, or 50-fold greater than) a threshold difference (e.g., threshold reduction) value derived from a cohort of at least 100 or at least 200 healthy test individuals in response to therapy administration can indicate a clinical or biochemical response to the GRA.
  • the cohort size is at least 20 or 30 or 50 test individuals.
  • the cohort can comprise or consist of subjects known or suspected of hav ing Cushing's syndrome.
  • the cohort of test individuals known or suspected of having Cushing's syndrome is an untreated control cohort.
  • the cohort can comprise or consist of subjects having Cushing's syndrome and exhibiting a clinical or biochemical response to an administered GRA or other medical or surgical therapy for treatment of Cushing's, e.g., as indicated by a reduction in, or elimination of, one or more symptoms of Cushing ' s.
  • a measured FKBP5 amount, activity, or expression level that is similar to (e.g., less than 5%, 10%, 25%, 30%>, 50%, 75%, or 100% above, or at least 95%, 90%, 80%, 70%, 60%, or 50% of) a threshold value derived from a cohort of 20 or 30 or 50 test individuals known or suspected of having (e.g. , untreated) Gushing 's syndrome can indicate that the subject has Gushing 's syndrome.
  • a measured FKBP5 amount, activity, or expression level that is lower than e.g.
  • a threshold value derived from a cohort of 20 or 30 or 50 test individuals known or suspected of having (e.g. , untreated) Gushing ' s syndrome can indicate that the subject does not have Cushing's syndrome.
  • a measured reduction in FKBP5 amount, activity, or expression level from pre-GRA administration to post-GRA administration that is similar to a threshold difference value derived from a cohort of 20 to 30, 20 to 50, 30 to 50, or at least 20, 30, or 50 individuals known or suspected of having Cushing's syndrome and exhibiting a clinical or biochemical response to GRA administration can indicate a clinical or biochemical response to the GILA.
  • the reduction from pre-GRA administration to post-GRA administration does not meet the threshold difference value derived from the cohort of 20 to 30, 20 to 50, 30 to 50, or at least 20, 30, or 50 individuals known or suspected of having Cushing's syndrome and exhibiting a clinical or biochemical response to GRA
  • the threshold, control, or threshold difference value derived from the cohort can be a contained in a database.
  • comparisons to the threshold, control, or threshold difference value can comprise querying the database.
  • the comparison further comprises applying a test of statistical significance.
  • the test of statistical significance includes a comparison of the relative variability of the amount, activity, or expression level in the cohort to the amount, activity, or expression level in the human subject.
  • the test of statistical significance includes a comparison of the relative variability in a change in FKBP5 amount, activit 7 , or expression from pre- to post- medical or surgical therapy administration or from pre- or post-GRA administration in the cohort to the change in the amount, activity, or expression level of FKBP5 in the human subject.
  • the methods include identifying whether the comparison is statistically significant in view of the inherent variability of the measurement as indicated by the observed variability in the cohort. iv. GRA administration
  • a GRA administered to the human subject can be any glucocorticoid receptor antagonist, including any of the GRAs described herein.
  • Exemplary GRAs include, but are not limited to, mifepristone, or a heteroaryl-ketone GRA, in some cases, the GRA administered to the human subject is not mifepristone.
  • the GRA is a specific GRA that preferentially binds to, and antagonizes, the GR ratlier than other nuclear receptors, such as mineralocorticoid receptor (MR), androgen receptor (AR), or progesterone receptor (PR).
  • MR mineralocorticoid receptor
  • AR androgen receptor
  • PR progesterone receptor
  • the GRA is a pan-specific GRA that preferentially binds to, and antagonizes, the GR and one or two other nuclear receptors selected from the group consisting of MR, AR, and PR. In some cases, the GRA is a non-specific GRA that binds to and antagonizes GR, MR, AR, and PR.
  • the GRA can be a steroidal GRA, such as a compound containing a modified Cortisol steroid backbone.
  • a steroidal GRA such as a compound containing a modified Cortisol steroid backbone.
  • Cortisol steroid backbone to create glucocorticoid antagonists include modifications of the 11- ⁇ hydroxy group and modification of the 17- ⁇ side chain (See, e. g., Lefebvre (1989) J. Steroid Biochem. 33: 557-563).
  • Steroidal GRAs can also include androgen-type steroid compounds as described in U.S. Pat. No. 5,929,058, and the compounds disclosed in U.S. Pat. Nos. 4,296,206; 4,386,085: 4,447,424: 4,477,445;
  • Such steroidal GRAs include cortexolone, dexamethasone-oxetanone, 19-nordeoxycorticosterone, 19-norprogesterone, cortisol-21 -mesylate; dexamethasone-21 -mesylate, 11 p>-(4-dimethylaminoethoxyphenyl) ⁇ 17a-propynyl- 17p ⁇ hydroxy ⁇ 4,9 ⁇ estradien-3-one (RU009), and 17 -hydroxy- 17a- 19-(4- methylphenyl)androsta-4,9(l l)-dien-3-one (RU044).
  • the GRA can be a non-steroidal GRA.
  • Non-steroidal GRAs do not share structural homology to, or are not modifications of, Cortisol.
  • Such compounds include synthetic mimetics and analogs of proteins, including partially peptidic, pseudopeptidic and non- peptidic molecular entities.
  • Non-steroidal GRA compounds also include glucocorticoid receptor antagonists having a cyclohexyl-pyrimidine backbone, a fused azadecalin backbone, a heteroaryl ketone fused azadecalin backbone, or an octahydro fused azadecalin backbone.
  • glucocorticoid receptor antagonists having a cyclohexyl-pyrimidine backbone include those described in U.S. Patent No. 8,685,973.
  • Exemplary glucocorticoid receptor antagonists having a fused azadecalin backbone include those described in U.S. Patent Nos. 7,928,237; and 8,461,172.
  • Exemplary glucocorticoid receptor antagonists having a heteroaryl ketone fused azadecalin backbone include those described in U.S. 2014/0038926.
  • Exemplar ⁇ ' glucocorticoid receptor antagonists having an octohydro fused azadecalin backbone include those described in U.S. Provisional Patent Appl. No. 61/908,333, entitled Octahydro Fused Azadecalin Glucocorticoid Receptor Modulators, Attorney Docket No. 85178-887884 (007800US), filed on November 25, 2013.
  • non-steroidal GRAs include the GR antagonist compounds disclosed in U.S. Pat. Nos. 5,696, 127; 6,051,573; and 6,570,020; the GR antagonist compounds disclosed in US Patent Application 20020077356, the glucocorticoid receptor antagonists disclosed in Bradley et al, J. Med. Chem.
  • WO 96/19458 which describes non-steroidal compounds which are high-affinity, highly selective antagonists for steroid receptors, such as 6-substituted-l,2-dihydro-N-protected-quinolines.
  • the GRAs can be delivered by any suitable means, including oral, parenteral and topical methods. Transdermal administration methods, by a topical route, can be formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols.
  • the pharmaceutical preparation is preferably in unit dosage form.
  • the preparation is subdivided into unit doses containing appropriate quantities of a GRA
  • the unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as pocketed tablets, capsules, and powders in vials or ampoules.
  • the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of any of these in packaged form.
  • the GRA can be co-administered with other agents.
  • Co-administration includes administering the GRA within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of the oilier agent.
  • Co-administration also includes administering simultaneously, approximately simultaneously (e.g., within about 1, 5, 10, 15, 20, or 30 minutes of each other), or sequentially in any order.
  • the GRA can each be administered once a day, or two, three, or more times per day so as to provide the preferred dosage level per day.
  • co-administration can be accomplished by co-formulation, i.e., preparing a single pharmaceutical composition including a GRA and any other agent.
  • the various components can be formulated separately.
  • Exemplary compounds, compositions, or active agents that can be co-administered with a GRA include, but are not limited to, another GRA, an agent that inhibits cellular proliferation, an anti -cancer chemotherapeutic, or an antibody.
  • the GRA, and any other agents can be present in any suitable amount, and can depend on various factors including, but not limited to, weight and age of the subject, state of the disease, etc.
  • Suitable dosage ranges include from about 0.1 mg to about 10,000 mg, or about 1 rng to about 1000 mg, or about 10 mg to about 750 rng, or about 25 mg to about 500 mg, or about 50 mg to about 250 mg. Suitable dosages also include about 1 mg, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900 or 1000 mg. If the GRA is mifepristone, treatment can be further understood by reference to U.S. Application No. 13/677,465, the disclosure of which is incorporated by reference in its entiret '.
  • composition can also contain other compatible therapeutic agents.
  • the therapeutic agents can be used in combination with one another, with other active agents known to be useful in antagonizing a glucocorticoid receptor, or with adjunctive agents that may not be effective alone, but may contribute to the efficacy of the active agent. b. Diagnosis of Cushing's syndrome
  • Cushing's syndrome can be diagnosed by detecting an amount or activity of FKBP5 protein or an expression le vel of a gene encoding FKBP5 and correlating the amount, activity, or expression level with a presence or absence, or likelihood, of Cushing's syndrome.
  • the method can include: a) measuring an amount, or activity of FKBP5 protein or an expression level of a gene encoding F BP5 in a sample from the subject; and b) identifying the subject as likely to be suffering from Cushing's syndrome when the amount, activity, or expression level is indicative of Cushing's syndrome.
  • the sample can be obtained by a variety of means.
  • the sample can be obtained by collecting a blood sample (e.g. , a sample of whole blood or a fraction thereof).
  • the sample can be obtained by scraping epithelial cells (e.g. , nasal epithelial cells) of a subject.
  • Samples include, but are not limited to samples of human cells and tissues, such as blood samples, cerebrospinal fluid samples, synovial tissue samples, synovial fluid samples, brain tissue samples, blood vessel samples, or tumor (e.g., pituitar ' adenoma) samples.
  • the sample can be extracted to obtain FKBP1 protein, or FKBP5 nucleic acid.
  • cells can be lysed, and the protein fraction obtained.
  • the lysate is further fractionated to purify a specified cellular compartment.
  • the cell lysate can be fractionated to obtain a cytosoiic protein fraction.
  • the cell lysate can be fractionated to obtain a nuclear or nucleolar protein fraction.
  • the protein fraction can be assayed for FKBP5 protein levels or activity.
  • the cells can be lysed, and a nucleic acid (e.g. , ni NA) fraction obtained.
  • the nucleic acid fraction can be assayed for expression of the FKBP5 gene.
  • the cell lysate, or protein fraction thereof can be purified using a variety of methods to obtain a fraction enriched for FKBP5 protein, or a portion thereof.
  • cells can be lysed and contacted with a chromatography medium under conditions suitable to preferentially bind contaminants or target protein. Where contaminants are preferentially bound, target protein can be collected as a flow through fraction and assayed further. Where target protein is bound, the chromatography medium can be washed and the target protein eluted.
  • cells can be lysed and contacted with a capture reagent (e.g., a capture antibody or aptamer) that specifically binds to the FKBP5 protein, or a portion thereof.
  • a capture reagent e.g., a capture antibody or aptamer
  • the capture reagent can be immobilized on a solid support.
  • the FKBP5 protein or portion thereof can be eluted from the capture reagent and then detected or quantified.
  • the F BP5 protein or portion thereof can be detected or quantified as a capture reagent-bound form.
  • the cell lysate, or nucleic acid fraction thereof can be purified using a variety of methods to obtain an fraction enriched for a transcript of a gene encoding FKBP5.
  • cells can be lysed and nucleic acids can be precipitated or otherwise purified from the lysate.
  • the nucleic acids can be purified by contacting the sample, or a fraction thereof, with a surface immobilized oligodT moiety to preferentially bind poiyadenylated mR A.
  • Nucleic acids can be subject to amplification, hybridization, polymerization, reverse transcription, or a combination thereof.
  • the amplification, hybridization, polymerization, or reverse transcription is target specific such that the gene encoding FKBP5, a transcript thereof, or a portion thereof i s specifically amplified, hybridized, or reverse transcribed.
  • the amplification, hybridization, polymerization, or reverse transcription is not target specific such that the sample is subject to whole genome or whole transcriptome hybridization, polymerization, or reverse transcription.
  • Whole genome or other non-specific hybridization, polymerization, or reverse transcription can be performed with the use of one or more degenerate primers or probes. After non-specific hybridization, polymerization, or reverse transcription, the FKBP5 gene expression can be specifically detected and/or quantified.
  • the amount or activity of FKBP5 polypeptide, or a portion thereof can be measured by a variety of methods known in the art.
  • an ELISA e.g., sandwich ELISA
  • the ELISA is a sandwich ELISA, in which F BP5 polypeptide, or a portion thereof, is immobilized by binding to an immobilized capture reagent (e.g., capture antibody or aptamer), and the immobilized polypeptide or portion thereof is detected with a detection reagent (e.g. , detection antibody or aptamer).
  • an immobilized capture reagent e.g., capture antibody or aptamer
  • activity of the FKBP5 polypeptide, or a portion thereof can be measured by contacting the sample, or a protein extract thereof, with a proline containing peptide substrate to measure the FKBP5 -mediated cis-trans prolyl isomerase activity of the sample.
  • the isomerizatson of the substrate can be measured using an cis-trans proline isomer sensitive enzyme, such as a protease.
  • chymotrypsin which has a high substrate specificity and catalytic efficiency (k cat /Km) against peptide substrates having a trans-proline at the P2 position and phenylalanine or tyrosine at the 1 position, but very little or no specificity or catalytic efficiency against such peptides containing a cis-proline at P2, can be used to measure prolyl isomerase activity.
  • an N-succinyl-Ala-Leu- c/s-Pro-Phe- >-nitroanilide substrate can be used in combination with chymotrypsin, which preferably cleaves the trans prolyl isomer of the substrate to assay a sample or extract thereof for FKBP5 prolyl isomerase activity.
  • the production of the trans isomer of the substrate by isomerase activity of the FKBP5 or portion thereof can be measured by detecting the digestion of the nitroanilide substrate by the chymotrypsin as described, e.g., in Fischer et al. Nature. 1989 Feb 2;337(6206):476-8.
  • activity of the FKBP5 polypeptide, or a portion thereof can be detected or quantified by measuring the amount of FKBP5 protein bound to GR in the sample. This can be performed, e.g., by purifying GR or FKBP5 protein under conditions suitable to preserve binding between GR and FKBP5 protein. The purification product can then be assayed for the presence, absence, or quantity of the cognate binding partner. For example, cytosolic GR can be purified and the presence of F BP5 protein detected in the purification product. As another example, FKBP5 protein can be purified and the presence of GR can be detected in the purification product.
  • this can be performed using a sandwich ELISA-type assay in which the immobilized capture reagent recognizes one member of the GR:FKBP5 protein complex and the detection reagent recognizes the other member of the GR FKBP5 protein complex.
  • expression of a gene encoding FKBP5 protein can be measured by reverse transcription of FKBP5 mRNA, or a portion thereof, and quantitative amplification of the reverse transcription product or a portion thereof.
  • the quantitative amplification can be performed using PGR (e.g., real time PGR) or other amplification techniques known in the art.
  • the amplification can be detected by, e.g., detecting incorporation of an intercalating dye into the amplification product, degradation of a quenched fluorescence hydrolysis probe, or binding of quenched molecular beacon.
  • Any measured amount activity or expression level in a sample can be normalized to a reference.
  • expression level can be normalized to total mRNA levels, or the expression level of a reference gene.
  • Suitable reference genes include, but are not limited to, GAPDH, , hypoxanthine phosphoribosyltransferase I (HRPT1), ribosomal protein large PI , or another housekeeping gene.
  • HRPT1 hypoxanthine phosphoribosyltransferase I
  • FKBP5 protein amount can be normalized to total protein levels, or the level of a reference gene product.
  • Suitable reference gene products include, but are not limited to, actin, tubulin, COX IV, HRPT1, GAPDH, or another housekeeping gene product.
  • the amount or activity of F BP5 protein or expression level of the gene encoding FKBP5 can be compared to a control value or threshold to indicate the presence or absence, or likelihood, of Gushing 's syndrome.
  • the control or threshold is a positive control, such that when the amount or activity of FKBP5 protein or expression level of the gene encoding FKBP5 is near, equal to, or higher than the control or threshold value, Cushing's syndrome is indicated.
  • the control or threshold is a negative control, such that when the amount or activity of FKBP5 protein or expression level of the gene encoding F BP5 is near, equal to, or below the threshold value, the absence of Cushing's syndrome is indicated.
  • Raw expression level data in the samples are normalized against the expression level of the glyceraldehyde phosphate dehydrogenase (GAPDH) gene in each sample.
  • GPDH glyceraldehyde phosphate dehydrogenase
  • the normalized expression level of the FKBP5 gene in the whole blood samples after administration of the GR agonist prednisone is from about 10 to 50 fold above the pre-administration level.
  • Administration of die GRAs mifepristone or CORT125134 causes a significant decrease in the fold change in FKBP5 expression.
  • Binder EB Salyakina D, Lichtoer P, Wochnik GM, Ising M, Putz B, Papiol S, Seaman S, Lucae S, Kohli MA, Nickel T, Kunzel HE, Fuchs B, Majer M, Pfennig A, Kem N, Brunner J, Modell S, Baghai T, Deiml T, Zill P, Bondy B, Rupprecht R, Messer T, Kohnlein O, Dabitz H, Bruckl T, Muller N, Pfister H, Lieb R, Mueller JC, Lohmussaar E, Strom TM, Bettecken T, Meitinger T, Uhr M, Rein T, Holsboer F, Muller-Myhsok B. Polymorphisms in FKBP5 are associated with increased recurrence of depressive episodes and rapid response to antidepressant treatment. Nature genetics 2004, 36: 1319-1325.
  • Binder EB Bradley RG, Liu W, Epstein MP, Deveau TC, Mercer KB, Tang Y, Gillespie CF, Hem CM, Nemeroff CB, Schwartz AC, Cubells IF, Ressler KJ.
  • Binder EB The role of FKBP5, a co-chaperone of the glucocorticoid receptor in the pathogenesis and therapy of affective and anxiety disorders.
  • MacKinnon DF Mondimore FM, Schweizer B, Bipolar disorder phenome group, NIMH genetks initiative bipolar disorder consortium, DePaulo JR Jr., Gershon ES, McMahon FJ, Potash JB. Family-based association of FKBP5 in bipolar disorder. Mol. Psychiatry, 2009, 14:261-268.
  • Meyneil A Levis B, McCrimmon R, Sherwin R, Musatov, S, Mobbs, CV.
  • hypothalamic FkbpS 1 induced by fasting, and elevated hypothalamic expression promotes obese phenotypes.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Epidemiology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pathology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • General Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • Endocrinology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Diabetes (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
PCT/US2016/033143 2015-05-18 2016-05-18 Methods for diagnosing and assessing treatment for cushing's syndrome Ceased WO2016187347A1 (en)

Priority Applications (13)

Application Number Priority Date Filing Date Title
KR1020177035970A KR20180008627A (ko) 2015-05-18 2016-05-18 쿠싱 증후군을 진단하고 이에 대한 치료를 평가하는 방법
IL309712A IL309712A (en) 2015-05-18 2016-05-18 Methods for diagnosing and determining treatment for Cushing's syndrome
KR1020247001601A KR102876869B1 (ko) 2015-05-18 2016-05-18 쿠싱 증후군을 진단하고 이에 대한 치료를 평가하는 방법
MX2017014857A MX2017014857A (es) 2015-05-18 2016-05-18 Metodos para diagnosticar y evaluar tratamiento para sindrome de cushing.
PH1/2017/502099A PH12017502099B1 (en) 2015-05-18 2016-05-18 Methods for diagnosing and assessing treatment for cushing's syndrome
CA2984623A CA2984623C (en) 2015-05-18 2016-05-18 Methods for diagnosing and assessing treatment for cushing's syndrome
US15/570,710 US10604807B2 (en) 2015-05-18 2016-05-18 Methods for diagnosing and assessing treatment for Cushing's syndrome
EP16797242.1A EP3297728A4 (en) 2015-05-18 2016-05-18 METHOD FOR DIAGNOSIS AND ASSESSMENT OF THE TREATMENT OF THE CUSHING SYNDROME
JP2017559803A JP6755264B2 (ja) 2015-05-18 2016-05-18 クッシング症候群についての処置を診断および評価するための方法
AU2016264321A AU2016264321B2 (en) 2015-05-18 2016-05-18 Methods for diagnosing and assessing treatment for Cushing's syndrome
IL255612A IL255612A (en) 2015-05-18 2017-11-12 Methods for diagnosing and assessing treatment for cushing's syndrome
US15/839,666 US11268145B2 (en) 2015-05-18 2017-12-12 Methods for treating Cushing's syndrome
US16/823,802 US20200216905A1 (en) 2015-05-18 2020-03-19 Methods for diagnosing and assessing treatment for cushing's syndrome

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201562163130P 2015-05-18 2015-05-18
US62/163,130 2015-05-18

Related Child Applications (3)

Application Number Title Priority Date Filing Date
US15/570,710 A-371-Of-International US10604807B2 (en) 2015-05-18 2016-05-18 Methods for diagnosing and assessing treatment for Cushing's syndrome
US15/839,666 Continuation US11268145B2 (en) 2015-05-18 2017-12-12 Methods for treating Cushing's syndrome
US16/823,802 Division US20200216905A1 (en) 2015-05-18 2020-03-19 Methods for diagnosing and assessing treatment for cushing's syndrome

Publications (1)

Publication Number Publication Date
WO2016187347A1 true WO2016187347A1 (en) 2016-11-24

Family

ID=57320595

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2016/033143 Ceased WO2016187347A1 (en) 2015-05-18 2016-05-18 Methods for diagnosing and assessing treatment for cushing's syndrome

Country Status (10)

Country Link
US (3) US10604807B2 (enExample)
EP (1) EP3297728A4 (enExample)
JP (3) JP6755264B2 (enExample)
KR (2) KR20180008627A (enExample)
AU (1) AU2016264321B2 (enExample)
CA (1) CA2984623C (enExample)
IL (2) IL309712A (enExample)
MX (1) MX2017014857A (enExample)
PH (1) PH12017502099B1 (enExample)
WO (1) WO2016187347A1 (enExample)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108389135A (zh) * 2018-03-21 2018-08-10 安徽理工大学 一种基于灰色关联模型的突水水源判别方法
WO2018160775A1 (en) * 2017-03-01 2018-09-07 Corcept Therapeutics, Inc. Concomitant administration of glucocorticoid receptor modulators and cyp3a inhibitors
US11285145B2 (en) 2020-05-27 2022-03-29 Corcept Therapeutics Incorporated Concomitant administration of glucocorticoid receptor modulator relacorilant and paclitaxel, a dual substrate of CYP2C8 and CYP3A4

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113194957A (zh) * 2018-12-21 2021-07-30 科塞普特治疗公司 通过施用糖皮质激素受体调节剂治疗库欣综合征中的高凝血病
EP4157275A4 (en) * 2020-05-27 2024-06-05 Corcept Therapeutics Incorporated CONCURRENT ADMINISTRATION OF GLUCOCORTICOID RECEPTOR MODULATOR RELACTORILANTS AND CYP2C8 SUBSTRATES
MX2022014923A (es) * 2020-05-27 2023-01-04 Corcept Therapeutics Inc Administracion concomitante del modulador del receptor de glucocorticoides relacorilant y sustratos cyp2c9.

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080118521A1 (en) * 2003-04-14 2008-05-22 Mira Susa Spring Gene expression associated with osteoblast differentiation
US20120039812A1 (en) * 2008-09-12 2012-02-16 Florian Holsboer Genes associated with posttraumatic-stress disorder (ptsd)
US20120094945A1 (en) * 2009-01-09 2012-04-19 The Regents Of The University Of California Discovery of Selective Glucocorticoid Receptor Modulators by Multiplexed Reporter Screening
WO2013177559A2 (en) * 2012-05-25 2013-11-28 Corcept Therapeutics, Inc. Heteroaryl-ketone fused azadecalin glucocorticoid receptor modulators
US20140170768A1 (en) * 2012-12-14 2014-06-19 Joel R. L. Ehrenkranz Method for monitoring and assessing pituitary function

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU747956B2 (en) * 1997-10-06 2002-05-30 Board Of Trustees Of The Leland Stanford Junior University Methods for treating psychosis associated with glucocorticoid related dysfunction
JP2004526425A (ja) * 2000-11-28 2004-09-02 ワイス 前立腺癌の診断および治療に有用なｆｋｂｐ核酸およびポリペプチドの発現分析
WO2013039916A1 (en) 2011-09-12 2013-03-21 The United States Of America, Represented By The Secretary, Dept. Of Health And Human Services Compositions for and methods of treatment and enhanced detection of non-pituitary tumors
JP2013090761A (ja) 2011-10-25 2013-05-16 Osaka City Univ 立体視内視鏡装置

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080118521A1 (en) * 2003-04-14 2008-05-22 Mira Susa Spring Gene expression associated with osteoblast differentiation
US20120039812A1 (en) * 2008-09-12 2012-02-16 Florian Holsboer Genes associated with posttraumatic-stress disorder (ptsd)
US20120094945A1 (en) * 2009-01-09 2012-04-19 The Regents Of The University Of California Discovery of Selective Glucocorticoid Receptor Modulators by Multiplexed Reporter Screening
WO2013177559A2 (en) * 2012-05-25 2013-11-28 Corcept Therapeutics, Inc. Heteroaryl-ketone fused azadecalin glucocorticoid receptor modulators
US20140170768A1 (en) * 2012-12-14 2014-06-19 Joel R. L. Ehrenkranz Method for monitoring and assessing pituitary function

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
GLAXOSMITHKLINE: "A Randomised, Double Blind, Placebo Controlled, 4 Period, Incomplete Block, Crossover Study Assessing the Doseresponse Curve of Fluticasone Propionate in an Antigen Challenge Chamber", CLINICAL TRIALS, 23 August 2012 (2012-08-23), pages 2, XP055331559, Retrieved from the Internet <URL:https://clinicaltrials.gov/ct2/show/NCT00848965> [retrieved on 20160713] *
LEE ET AL.: "A measure of glucocorticoid load provided by DNA methylation of Fkbp5 in mice.", PSYCHOPHARMACOLOGY, vol. 218, no. 1, 2011, pages 303 - 312, XP019963307 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018160775A1 (en) * 2017-03-01 2018-09-07 Corcept Therapeutics, Inc. Concomitant administration of glucocorticoid receptor modulators and cyp3a inhibitors
US10195214B2 (en) 2017-03-01 2019-02-05 Corcept Therapeutics, Inc. Concomitant administration of glucocorticoid receptor modulators and CYP3A inhibitors
US10842800B2 (en) 2017-03-01 2020-11-24 Corcept Therapeutics, Inc. Concomitant administration of glucocorticoid receptor modulators and CYP3A inhibitors
US11969435B2 (en) 2017-03-01 2024-04-30 Corcept Therapeutics, Inc. Concomitant administration of glucocorticoid receptor modulators and CYP3A inhibitors
US12097210B2 (en) 2017-03-01 2024-09-24 Corcept Therapeutics, Inc. Concomitant administration of glucocorticoid receptor modulators and CYP3A inhibitors
CN108389135A (zh) * 2018-03-21 2018-08-10 安徽理工大学 一种基于灰色关联模型的突水水源判别方法
US11285145B2 (en) 2020-05-27 2022-03-29 Corcept Therapeutics Incorporated Concomitant administration of glucocorticoid receptor modulator relacorilant and paclitaxel, a dual substrate of CYP2C8 and CYP3A4
US11944617B2 (en) 2020-05-27 2024-04-02 Corcept Therapeutics Incorporated Concomitant administration of glucocorticoid receptor modulator relacorilant and paclitaxel, a dual substrate of CYP2C8 and CYP3A4
US12589094B2 (en) 2020-05-27 2026-03-31 Corcept Therapeutics Incorporated Concomitant administration of glucocorticoid receptor modulator relacorilant and paclitaxel, a dual substrate of CYP2C8 and CYP3A4

Also Published As

Publication number Publication date
KR20180008627A (ko) 2018-01-24
EP3297728A4 (en) 2019-07-03
JP2020195401A (ja) 2020-12-10
KR102876869B1 (ko) 2025-10-27
JP6755264B2 (ja) 2020-09-16
AU2016264321B2 (en) 2021-02-18
US11268145B2 (en) 2022-03-08
US20180100179A1 (en) 2018-04-12
PH12017502099A1 (en) 2018-05-07
CA2984623A1 (en) 2016-11-24
US10604807B2 (en) 2020-03-31
AU2016264321A1 (en) 2017-11-30
KR20240014584A (ko) 2024-02-01
PH12017502099B1 (en) 2023-08-30
EP3297728A1 (en) 2018-03-28
JP2020079799A (ja) 2020-05-28
JP2018524562A (ja) 2018-08-30
US20200216905A1 (en) 2020-07-09
IL309712A (en) 2024-02-01
IL255612A (en) 2018-01-31
US20180291451A1 (en) 2018-10-11
MX2017014857A (es) 2018-08-15
CA2984623C (en) 2023-05-02
JP6893264B2 (ja) 2021-06-23

Similar Documents

Publication Publication Date Title
US11268145B2 (en) Methods for treating Cushing&#39;s syndrome
Gubelmann et al. Identification of the transcription factor ZEB1 as a central component of the adipogenic gene regulatory network
EP3079475B1 (en) Glucocorticoid inhibitors for treatment of prostate cancer
Padgett et al. Deoxyhypusine synthase, an essential enzyme for hypusine biosynthesis, is required for proper exocrine pancreas development
WO2007145992A2 (en) Genetic basis of treatment response in depression patients
Nakata et al. Epigenetic upregulation of Schlafen11 renders WNT-and SHH-activated medulloblastomas sensitive to cisplatin
van Zyl et al. Isoginkgetin leads to decreased protein synthesis and activates an ATF4-dependent transcriptional response
Zheng et al. Stress induces corticosterone-mediated CD8+ T cell exhaustion to promote non-small cell lung cancer
EP4073521A2 (en) Materials and methods for monitoring inflammation
Müller et al. SNP dependent modulation of circulating miRNAs from the miR25/93/106 cluster in patients undergoing weight loss
Gilad et al. Steroid receptor coactivators: from basic research to translational opportunities
CN112795651B (zh) Muc20作为诊断多发性套细胞淋巴瘤蛋白酶体抑制剂耐药的标志物及其应用
Cucuzza et al. Role of FKBP51 in the modulation of the expression of the corticosteroid receptors in bovine thymus following glucocorticoid administration
JP2015159766A (ja) クッシング症候群の検査方法、検査用バイオマーカー及び治療剤
WO2009045403A2 (en) MENIN REGULATION OF β-ISLET CELL PROLIFERATION
Olczak et al. Glucocorticoid receptor (NR3C1) gene polymorphisms are associated with age and blood parameters in Polish Caucasian nonagenarians and centenarians
US20070254303A1 (en) Methods for assessing the efficacy of treatment with a glucocorticoid
EP3255433A1 (en) Methods using blm as a marker of multiple myeloma
WO2015089311A1 (en) Sgk1 inhibitors for treatment of prostate cancer
CN105008550A (zh) 包含对muc4基因多态性进行检测的步骤的、判定抗癌剂疗法的副作用的发生风险的方法
Karakuz et al. EXPERIMENTAL BIOMEDICAL RESEARCH
Karakuz et al. Is there a relationship between polycystic ovary syndrome and the FABP1 gene rs2197076 single nucleotide polymorphism?
Sánchez Pascua Carboxylesterase 1 genetic variability, expression and potential for drug-drug interations
Hynne Glucocorticoid pharmacogenetics in Addison’s disease–The role of the immunophilin FK506-binding protein (FKBP51) for glucocorticoid sensitivity
Nelson et al. Changmeng Cai, Housheng Hansen He, 2, 3 Sen Chen, Ilsa Coleman, 4 Hongyun Wang, Zi Fang, Shaoyong Chen

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16797242

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2984623

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: 255612

Country of ref document: IL

ENP Entry into the national phase

Ref document number: 2017559803

Country of ref document: JP

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 11201709449U

Country of ref document: SG

WWE Wipo information: entry into national phase

Ref document number: MX/A/2017/014857

Country of ref document: MX

Ref document number: 12017502099

Country of ref document: PH

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2016264321

Country of ref document: AU

Date of ref document: 20160518

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 20177035970

Country of ref document: KR

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 2016797242

Country of ref document: EP