+

WO2016007535A1 - Antagonistes de hsp90/cdc37 et méthodes d'utilisation correspondantes - Google Patents

Antagonistes de hsp90/cdc37 et méthodes d'utilisation correspondantes Download PDF

Info

Publication number
WO2016007535A1
WO2016007535A1 PCT/US2015/039415 US2015039415W WO2016007535A1 WO 2016007535 A1 WO2016007535 A1 WO 2016007535A1 US 2015039415 W US2015039415 W US 2015039415W WO 2016007535 A1 WO2016007535 A1 WO 2016007535A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
substituted
hexamethyl
hydroxy
carboxamide
Prior art date
Application number
PCT/US2015/039415
Other languages
English (en)
Inventor
Mei-Sze Chua
Wei Wei
Song Wu
Samuel So
Original Assignee
The Board Of Trustees Of The Leland Stanford Junior University
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
Application filed by The Board Of Trustees Of The Leland Stanford Junior University filed Critical The Board Of Trustees Of The Leland Stanford Junior University
Publication of WO2016007535A1 publication Critical patent/WO2016007535A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
    • C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids

Definitions

  • Hepatocellular carcinoma the most common adult liver malignancy, is the seventh most common cancer and the second most frequent cause of cancer-related death worldwide.
  • Most of the burden (80%) of HCC is borne in the developing world, such as Eastern and Southeast Asia and sub-Saharan Africa, where the dominant risk factor is chronic infection with hepatitis B virus (HBV), together with exposure with aflatoxin B1 .
  • HBV hepatitis B virus
  • the dominant risk factor is chronic infection with hepatitis C virus.
  • HCC has a poor prognosis, partly due to late diagnosis of the disease and lack of effective therapeutic options. Most patients remain asymptomatic until the disease is advanced. Although more than 50 drugs that target different biomarkers or signaling pathways are in clinical trials for HCC treatment, as yet there is no therapeutic agent superior to sorafenib, a tyrosine kinase inhibitor, which was FDA approved as the standard of care for advanced HCC. Due to recent emergence of resistance to sorafenib, second-line therapies with other targets are highly desirable.
  • the chaperone-kinome pathway is attractive as a therapeutic target in cancer.
  • the heat shock protein 90 (HSP90) cooperates with its molecular co-chaperone CDC37 to regulate the folding, maturation, stabilization, and phosphorylation of a wide array of protein kinases, which are important mediators of signal transduction and cell growth in human cancers.
  • HSP90 has been recognized as a key facilitator of oncogene addiction and a promising therapeutic target in cancers, with several HSP90 inhibitors in preclinical and clinical evaluation for cancer therapy.
  • Current HSP90 inhibitors interact with the N-terminal ATP-binding pocket and block ATP binding to stop the chaperone cycle, thereby leading to client protein
  • CDC37 is the key permissive factor in cell transformation caused by oncogenic protein kinases.
  • the central segment of CDC37 associates with the N-terminal ATPase domain of HSP90, and the N-terminal segment of CDC37 associates with its client protein kinases (e.g., Cdk4, EGFR, AKT, MEK1/2 and Raf family proteins).
  • the present disclosure provides compounds and methods of use in the treatment of cancer (e.g, hepatocellular carcinoma).
  • cancer e.g, hepatocellular carcinoma
  • This disclosure concerns compounds which are useful as HSP90/CDC37 antagonists and are thus useful for treating a variety of diseases and disorders that are mediated or sustained through the interaction of HSP90 and CDC37.
  • This disclosure also relates to pharmaceutical compositions that include these compounds, including formulations providing an effective dose, e.g. a unit dose, of the compound for treatment of cancer, including without limitation carcinomas; methods of using these compounds in the treatment of various diseases and disorders, and processes for preparing these compounds.
  • X is selected from:
  • aspects of the present disclosure include a compound, and formulations thereof, selected from:
  • aspects of the present disclosure include a pharmaceutical composition
  • a pharmaceutical composition comprising a compound as described above and a pharmaceutically acceptable carrier, e.g. in a unit dose effective for treating cancer.
  • aspects of the present disclosure include a method of antagonizing the interaction of HSP90 with CDC37 in a biological sample or a patient, which method can include contacting the biological sample or administering to the patient a compound as described above.
  • antagonizing the interaction of HSP90 with CDC37 results in treatment of a disease or disorder that is mediated or sustained through the interaction of HSP90 with CDC37.
  • the disease or disorder is cancer (e.g., a carcinoma such as hepatocellular carcinoma).
  • aspects of the present disclosure include a method of antagonizing the interaction of HSP90 with CDC37 in an individual having cancer, which method can include administering to the individual, at a dose effective for antagonizing the interaction of HSP90 with CDC37 in cancer cells, a compound as described above.
  • the cancer is a carcinoma (e.g., hepatocellular carcinoma).
  • aspects of the present disclosure include a method of treating an individual having cancer, which method can include administering to the individual, at a dose effective for reducing the number of cancer cells in the individual, a compound as described above.
  • the cancer is a carcinoma (e.g., hepatocellular carcinoma).
  • Such a method may comprise (i) identifying a patient having hepatocellular cancer; and administering to the individual an effective dose of a compound described above, optionally in combination with additional chemotherapeutic agent(s).
  • Another aspect of the present invention relates to the use of a compound described above in the manufacture of a medicament for the treatment of cancer, including hepatocellular carcinoma, wherein the medicament is administered to a patient in an effective dose, optionally in combination with additional chemotherapeutic agent(s).
  • kits for treatment of cancer includes a compound described herein, in an amount sufficient to reduce the growth of the cancer cells.
  • aspects of the present disclosure further include a method for making Compound 2, Compound 3, or Compound 4, where the method includes contacting
  • FIG. 1 Celastrol and its derivatives disrupt HSP90/CDC37 interaction in HCC cells. HepG2 cells were incubated with 5 ⁇ of each compound and the same volume of DMSO as negative control for 6 hours. HSP90/CDC37 complex was then pulled down by anti-CDC37 antibody in the cell lysates. Anti-HSP90 antibody was used to detect the HSP90 protein in the complex. The lysates were used to detect HSP90, CDC37, and GAPDH (loading control).
  • FIGS 2A-C Figures 2A-C.
  • Celastrol and its derivatives are preferentially inhibited viability of HCC cells compared to normal hepatocytes.
  • A CDC37 and GAPDH (loading control) expression were determined by Western Blot using specific antibodies in HCC cells (HepG2, Huh7, and Hep3B) and normal hepatocytes (Hu81 14, Hu4175, and Hu8130).
  • B Phase-contrast microscopic examination of the effect of cel-D7 (5 ⁇ ) on HepG2 cells and normal hepatocytes Hu4175.
  • C Celastrol and its derivatives (5 ⁇ each) induced apoptosis in Huh7 cells after 6 hours treatment. Cells were stained with TUNEL and DAPI as described under Materials and Methods to detect for apoptotic cells. Fluorescence labeling was visualized and photographed at 100x magnification.
  • FIGS 3A-C Celastrol and its derivatives induced degradation and inhibited phosphorylation of HSP90/CDC37 client proteins in HCC cell lines. HepG2, Huh7, and Hep3B cells were incubated for 6 hours with each compound (at 1 or 10 ⁇ ) and CDC37, HSP90/CDC37 client proteins and GAPDH (loading control) levels were determined by Western blotting using specific antibodies.
  • FIGS 4A-F Celastrol and its derivatives inhibited growth of orthotopic HCC patient- derived xenografts.
  • A CDC37 and GAPDH (loading control) protein expressions in the tumor (T) and matched non-tumor liver (N) tissues of three HCC patients were determined by Western Blot.
  • E E).
  • FIG. 1 Celastrol and its derivatives induced apoptosis (TUNEL Assay) in orthotopic HCC patient-derived xenografts. Representative images are shown for HCC-3 xeongrafts after 3 weeks treatment of each compound and saline control (200x magnification). Apoptotic cells are defined by cells with brown nucleic staining..
  • Figure 7 presents a table of structures and activities of celastrol and subject
  • FIGS 8A-D Celastrol and its derivatives preferentially inhibited viability of HCC cells compared to normal hepatocytes.
  • Cell viability assays based on ATP release were used to determine the cytotoxicity of celastrol and its derivatives on three human HCC cell lines cells (HepG2, Huh7, and Hep3B) and normal hepatocytes (Hu81 14, Hu4175, and Hu8130) following 72 hours of treatment. Results are presented as mean ⁇ SD (error bars). Relative ATP activity is proportional to the number of viable cells.
  • the values of luciferase activity were normalized and compared with the DMSO control value, which was set at 100% cell viability. Three independent experiments were done, each in triplicates.
  • Figures 9A-F depict bioluminescence images of three HCC patient-derived xenograft models (mice) shown before treatment and after 3 weeks of treatment with celastrol or its derivatives.
  • Orthotopic liver tumor models derived from human HCC patient specimens (HCC-1 , HCC-2, HCC-3) expressing a trifusion reporter gene were given intravenous injection of celastrol or its derivatives, and the tumor growth was monitored weekly using the Xenogen MS 100 imaging system.
  • FIGS 10A-B depict synthetic schemes for (A) cel-D2 and (B) cel-D7.
  • DIPEA DIPEA
  • Figure 11 depicts 1 H NMR data for cel-D2
  • Figure 12 depicts 13 C NMR data for cel-D7
  • Figure 13 depicts 13 C NMR data for cel-D2
  • Figure 14 depicts 1 H NMR data for cel-D7
  • Figure 15 depicts high resolution mass spectrometry data for cel-D2
  • Figure 16 depicts high resolution mass spectrometry data for cel-D7
  • This disclosure concerns compounds which are useful as HSP90/CDC37 antagonists and are thus useful for treating a variety of diseases and disorders that are mediated or sustained through the interaction of HSP90 and CDC37.
  • This disclosure also relates to pharmaceutical compositions that include these compounds, methods of using these compounds in the treatment of various diseases and disorders, and processes for preparing these compounds.
  • dates of publication provided may be different from the actual publication dates which may need to be independently confirmed
  • Alkyl refers to monovalent saturated aliphatic hydrocarbyl groups having from 1 to 10 carbon atoms and preferably 1 to 6 carbon atoms. This term includes, by way of example, linear and branched hydrocarbyl groups such as methyl (CH 3 -), ethyl (CH 3 CH 2 -), n-propyl (CH 3 CH 2 CH 2 -), isopropyl ((CH 3 ) 2 CH-), n-butyl (CH 3 CH 2 CH 2 CH 2 -), isobutyl ((CH 3 ) 2 CHCH 2 -), sec- butyl ((CH 3 )(CH 3 CH 2 )CH-), t-butyl ((CH 3 ) 3 C-), n-pentyl (CH 3 CH 2 CH 2 CH 2 CH 2 -), and neopentyl ((CH 3 ) 3 CCH 2 -).
  • substituted alkyl refers to an alkyl group as defined herein wherein one or more carbon atoms in the alkyl chain have been optionally replaced with a heteroatom such as -0-, -N-, -S-, -S(0) n - (where n is 0 to 2), -NR- (where R is hydrogen or alkyl) and having from 1 to 5 substituents selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy
  • Alkylene refers to divalent aliphatic hydrocarbyl groups preferably having from 1 to 6 and more preferably 1 to 3 carbon atoms that are either straight-chained or branched, and which are optionally interrupted with one or more groups selected from -0-, -NR 10 -, -NR 10 C(O)-, -C(0)NR 10 - and the like.
  • This term includes, by way of example, methylene (-CH 2 -), ethylene (-CH 2 CH 2 -), n-propylene (-CH 2 CH 2 CH 2 -), iso-propylene (-CH 2 CH(CH 3 )-), (-C(CH 3 ) 2 CH 2 CH 2 -), (-C(CH 3 ) 2 CH 2 C(0)-), (-C(CH 3 ) 2 CH 2 C(0)NH-), (-CH(CH 3 )CH 2 -), and the like.
  • Substituted alkylene refers to an alkylene group having from 1 to 3 hydrogens replaced with substituents as described for carbons in the definition of “substituted” below.
  • alkane refers to alkyl group and alkylene group, as defined herein.
  • alkylaminoalkyl refers to the groups R NHR - where R is alkyl group as defined herein and R is alkylene, alkenylene or alkynylene group as defined herein.
  • alkaryl or "aralkyl” refers to the groups -alkylene-aryl and -substituted alkylene-aryl where alkylene, substituted alkylene and aryl are defined herein.
  • Alkoxy refers to the group -O-alkyl, wherein alkyl is as defined herein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, sec- butoxy, n-pentoxy, and the like.
  • alkoxy also refers to the groups alkenyl-O-, cycloalkyl-O-, cycloalkenyl-O-, and alkynyl-O-, where alkenyl, cycloalkyi, cycloalkenyl, and alkynyl are as defined herein.
  • substituted alkoxy refers to the groups substituted alkyl-O-, substituted alkenyl-O-, substituted cycloalkyl-O-, substituted cycloalkenyl-O-, and substituted alkynyl-O- where substituted alkyl, substituted alkenyl, substituted cycloalkyi, substituted cycloalkenyl and substituted alkynyl are as defined herein.
  • alkoxyamino refers to the group -NH-alkoxy, wherein alkoxy is defined herein.
  • haloalkoxy refers to the groups alkyl-O- wherein one or more hydrogen atoms on the alkyl group have been substituted with a halo group and include, by way of examples, groups such as trifluoromethoxy, and the like.
  • haloalkyl refers to a substituted alkyl group as described above, wherein one or more hydrogen atoms on the alkyl group have been substituted with a halo group.
  • groups include, without limitation, fluoroalkyl groups, such as trifluoromethyl, difluoromethyl, trifluoroethyl and the like.
  • alkylalkoxy refers to the groups -alkylene-O-alkyl, alkylene-O-substituted alkyl, substituted alkylene-O-alkyl, and substituted alkylene-O-substituted alkyl wherein alkyl, substituted alkyl, alkylene and substituted alkylene are as defined herein.
  • alkylthioalkoxy refers to the group -alkylene-S-alkyl, alkylene-S-substituted alkyl, substituted alkylene-S-alkyl and substituted alkylene-S-substituted alkyl wherein alkyl, substituted alkyl, alkylene and substituted alkylene are as defined herein.
  • Alkenyl refers to straight chain or branched hydrocarbyl groups having from 2 to 6 carbon atoms and preferably 2 to 4 carbon atoms and having at least 1 and preferably from 1 to 2 sites of double bond unsaturation. This term includes, by way of example, bi-vinyl, allyl, and but-3-en-1-yl. Included within this term are the cis and trans isomers or mixtures of these isomers.
  • substituted alkenyl refers to an alkenyl group as defined herein having from 1 to 5 substituents, or from 1 to 3 substituents, selected from alkoxy, substituted alkoxy, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,
  • Alkynyl refers to straight or branched monovalent hydrocarbyl groups having from 2 to
  • alkynyl groups include acetylenyl (-C ⁇ CH), and propargyl (-CH 2 C ⁇ CH).
  • substituted alkynyl refers to an alkynyl group as defined herein having from 1 to 5 substituents, or from 1 to 3 substituents, selected from alkoxy, substituted alkoxy, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,
  • Alkynyloxy refers to the group -O-alkynyl, wherein alkynyl is as defined herein.
  • Alkynyloxy includes, by way of example, ethynyloxy, propynyloxy, and the like.
  • Acyl refers to the groups H-C(O)-, alkyl-C(O)-, substituted alkyl-C(O)-, alkenyl-C(O)-, substituted alkenyl-C(O)-, alkynyl-C(O)-, substituted alkynyl-C(O)-, cycloalkyl-C(O)-, substituted cycloalkyl-C(O)-, cycloalkenyl-C(O)-, substituted cycloalkenyl-C(O)-, aryl-C(O)-, substituted aryl-C(O)-, heteroaryl-C(O)-, substituted heteroaryl-C(O)-, heterocyclyl-C(O)-, and substituted heterocyclyl-C(O)-, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, substitute
  • Acylamino refers to the groups -NR 20 C(O)alkyl, -NR 20 C(O)substituted alkyl, N
  • R 20 C(O)cycloalkyl, -NR 20 C(O)substituted cycloalkyi, - NR U C(0)cycloalkenyl, -NR U C(0)substituted cycloalkenyl, -NFTC(0)alkenyl, - NR 20 C(O)substituted alkenyl, -NR 20 C(O)alkynyl, -NR 20 C(O)substituted
  • Aminocarbonyl or the term “aminoacyl” refers to the group -C(0)NR 21 R 22 , wherein R 21 and R 22 independently are selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic and where R 21 and R 22 are optionally joined together with the nitrogen bound thereto to form a heterocyclic or substituted heterocyclic group, and wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl
  • Aminocarbonylamino refers to the group -NR 21 C(0)NR 22 R 23 where R 21 , R 22 , and R 23 are independently selected from hydrogen, alkyl, aryl or cycloalkyi, or where two R groups are joined to form a heterocyclyl group.
  • alkoxycarbonylamino refers to the group -NRC(0)OR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclyl wherein alkyl, substituted alkyl, aryl, heteroaryl, and heterocyclyl are as defined herein.
  • acyloxy refers to the groups alkyl-C(0)0-, substituted alkyl-C(0)0-, cycloalkyl-C(0)0-, substituted cycloalkyl-C(0)0-, aryl-C(0)0-, heteroaryl-C(0)0-, and heterocyclyl-C(0)0- wherein alkyl, substituted alkyl, cycloalkyi, substituted cycloalkyi, aryl, heteroaryl, and heterocyclyl are as defined herein.
  • Aminosulfonyl refers to the group -S0 2 NR 21 R 22 , wherein R 21 and R 22 independently are selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic and where R 21 and R 22 are optionally joined together with the nitrogen bound thereto to form a heterocyclic or substituted heterocyclic group and alkyl, substituted alkyi, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, aryl, substituted substituted
  • “Sulfonylamino” refers to the group -NR 21 S0 2 22 , wherein R 21 and R 22 independently are selected from the group consisting of hydrogen, alkyi, substituted alkyi, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic and where R 21 and R 22 are optionally joined together with the atoms bound thereto to form a heterocyclic or substituted heterocyclic group, and wherein alkyi, substituted alkyi, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, aryl
  • Aryl or “Ar” refers to a monovalent aromatic carbocyclic group of from 6 to 18 carbon atoms having a single ring (such as is present in a phenyl group) or a ring system having multiple condensed rings (examples of such aromatic ring systems include naphthyl, anthryl and indanyl) which condensed rings may or may not be aromatic, provided that the point of attachment is through an atom of an aromatic ring. This term includes, by way of example, phenyl and naphthyl.
  • such aryl groups can optionally be substituted with from 1 to 5 substituents, or from 1 to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyi, alkoxy, alkenyl, alkynyl, cycloalkyi, cycloalkenyl, substituted alkyi, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyi, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl, carboxylalkyi, cyano, halogen, nitro, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thi
  • Aryloxy refers to the group -O-aryl, wherein aryl is as defined herein, including, by way of example, phenoxy, naphthoxy, and the like, including optionally substituted aryl groups as also defined herein.
  • Amino refers to the group -NH 2 .
  • substituted amino refers to the group -NRR where each R is independently selected from the group consisting of hydrogen, alkyi, substituted alkyi, cycloalkyi, substituted cycloalkyi, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl, and heterocyclyl provided that at least one R is not hydrogen.
  • azido refers to the group -N 3 .
  • Carboxyl refers to -C0 2 H or salts thereof.
  • Carboxyl ester or “carboxy ester” or the terms “carboxyalkyl” or “carboxylalkyl” refers to the groups -C(0)0-alkyl, -C(0)0-substituted alkyl, -C(0)0-alkenyl, -C(0)0-substituted alkenyl, -C(0)0-alkynyl, -C(0)0-substituted alkynyl, -C(0)0-aryl, -C(0)0-substituted aryl, -C(0)0-cycloalkyl, -C(0)0-substituted cycloalkyi, -C(0)0-cycloalkenyl, -C(0)0-substituted cycloalkenyl, -C(0)0-heteroaryl, -C(0)0-substituted heteroaryl, -C(0)0-heterocyclic, and -C(0)0-substituted heterocyclic, wherein alkyl, substitute
  • (Carboxyl ester)oxy refers to the groups -0-C(0)0- alkyl, -0-C(0)0-substituted alkyl, -0-C(0)0-alkenyl, -0-C(0)0-substituted alkenyl, -0-C(0)0- alkynyl, -0-C(0)0-substituted alkynyl, -0-C(0)0-aryl, -0-C(0)0-substituted aryl, -0-C(0)0- cycloalkyl, -0-C(0)0-substituted cycloalkyi, -0-C(0)0-cycloalkenyl, -0-C(0)0-substituted cycloalkenyl, -0-C(0)0-heteroaryl, -0-C(0)0-substituted heteroaryl, -0-C(0)0-heterocyclic, and -0-C(0)0-substituted heterocyclic, wherein alkyl, substituted alkyl
  • Cycloalkyi refers to cyclic alkyl groups of from 3 to 10 carbon atoms having single or multiple cyclic rings including fused, bridged, and spiro ring systems.
  • suitable cycloalkyi groups include, for instance, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl and the like.
  • Such cycloalkyi groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, and the like.
  • substituted cycloalkyi refers to cycloalkyi groups having from 1 to 5 substituents, or from 1 to 3 substituents, selected from alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamin
  • Cycloalkenyl refers to non-aromatic cyclic alkyl groups of from 3 to 10 carbon atoms having single or multiple rings and having at least one double bond and preferably from 1 to 2 double bonds.
  • substituted cycloalkenyl refers to cycloalkenyl groups having from 1 to 5 substituents, or from 1 to 3 substituents, selected from alkoxy, substituted alkoxy, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro
  • Cycloalkynyl refers to non-aromatic cycloalkyi groups of from 5 to 10 carbon atoms having single or multiple rings and having at least one triple bond.
  • Cycloalkoxy refers to -O-cycloalkyl
  • Cycloalkenyloxy refers to -O-cycloalkenyl.
  • Halo or halogen refers to fluoro, chloro, bromo, and iodo.
  • Heteroaryl refers to an aromatic group of from 1 to 15 carbon atoms, such as from 1 to 10 carbon atoms and 1 to 10 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur within the ring.
  • Such heteroaryl groups can have a single ring (such as, pyridinyl, imidazolyl or furyl) or multiple condensed rings in a ring system (for example as in groups such as, indolizinyl, quinolinyl, benzofuran, benzimidazolyl or benzothienyl), wherein at least one ring within the ring system is aromatic and at least one ring within the ring system is aromatic , provided that the point of attachment is through an atom of an aromatic ring.
  • the nitrogen and/or sulfur ring atom(s) of the heteroaryl group are optionally oxidized to provide for the N-oxide (N ⁇ 0), sulfinyl, or sulfonyl moieties.
  • This term includes, by way of example, pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl.
  • heteroaryl groups can be optionally substituted with 1 to 5 substituents, or from 1 to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyi, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyi, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl, carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thio
  • heteroaryl refers to the groups -alkylene-heteroaryl where alkylene and heteroaryl are defined herein. This term includes, by way of example, pyridylmethyl, pyridylethyl, indolylmethyl, and the like.
  • Heteroaryloxy refers to -O-heteroaryl.
  • Heterocycle refers to a saturated or unsaturated group having a single ring or multiple condensed rings, including fused bridged and spiro ring systems, and having from 3 to 20 ring atoms, including 1 to 10 hetero atoms.
  • These ring atoms are selected from the group consisting of nitrogen, sulfur, or oxygen, wherein, in fused ring systems, one or more of the rings can be cycloalkyi, aryl, or heteroaryl, provided that the point of attachment is through the non-aromatic ring.
  • the nitrogen and/or sulfur atom(s) of the heterocyclic group are optionally oxidized to provide for the N-oxide, -S(O)-, or -S0 2 - moieties.
  • heterocycles and heteroaryls include, but are not limited to, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,
  • phenanthridine acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1 ,2,3,4- tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene, benzo[b]thiophene, morpholinyl, thiomorpholinyl (also referred to as thiamorpholinyl), 1 ,1 - dioxothiomorpholinyl, piperidinyl, pyrrolidine, tetrahydrofuranyl, and the like.
  • heterocyclic groups can be optionally substituted with 1 to 5, or from 1 to 3 substituents, selected from alkoxy, substituted alkoxy, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro,
  • Heterocyclyloxy refers to the group -O-heterocyclyl.
  • heterocyclylthio refers to the group heterocyclic-S-.
  • heterocyclene refers to the diradical group formed from a heterocycle, as defined herein.
  • hydroxyamino refers to the group -NHOH.
  • Niro refers to the group -N0 2 .
  • “Sulfonyl” refers to the group S0 2 -alkyl, S0 2 -substituted alkyl, S0 2 -alkenyl, S0 2 - substituted alkenyl, S0 2 -cycloalkyl, S0 2 -substituted cylcoalkyi, S0 2 -cycloalkenyl, S0 2 - substituted cylcoalkenyl, S0 2 -aryl, S0 2 -substituted aryl, S0 2 -heteroaryl, S0 2 -substituted heteroaryl, S0 2 -heterocyclic, and S0 2 -substituted heterocyclic, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substituted
  • “Sulfonyloxy” refers to the group -OS0 2 -alkyl, OS0 2 -substituted alkyl, OS0 2 -alkenyl, OS0 2 -substituted alkenyl, OS0 2 -cycloalkyl, OS0 2 -substituted cylcoalkyi, OS0 2 -cycloalkenyl, OS0 2 -substituted cylcoalkenyl, OS0 2 -aryl, OS0 2 -substituted aryl, OS0 2 -heteroaryl, OS0 2 - substituted heteroaryl, OS0 2 -heterocyclic, and OS0 2 substituted heterocyclic, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyi, substituted cycloalkyi, cycloalkenyl, substitute
  • aminocarbonyloxy refers to the group -OC(0)NRR where each R is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl, or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic are as defined herein.
  • Thiol refers to the group -SH.
  • Alkylthio or the term “thioalkoxy” refers to the group -S-alkyl, wherein alkyl is as defined herein.
  • sulfur may be oxidized to -S(O)-.
  • the sulfoxide may exist as one or more stereoisomers.
  • substituted thioalkoxy refers to the group -S-substituted alkyl.
  • thioaryloxy refers to the group aryl-S- wherein the aryl group is as defined herein including optionally substituted aryl groups also defined herein.
  • heteroaryloxy refers to the group heteroaryl-S- wherein the heteroaryl group is as defined herein including optionally substituted aryl groups as also defined herein.
  • thioheterocyclooxy refers to the group heterocyclyl-S- wherein the
  • heterocyclyl group is as defined herein including optionally substituted heterocyclyl groups as also defined herein.
  • substituted when used to modify a specified group or radical, can also mean that one or more hydrogen atoms of the specified group or radical are each, independently of one another, replaced with the same or different substituent groups as defined below.
  • R 60 is selected from the group consisting of optionally substituted alkyl, cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl, each R 70 is independently hydrogen or R 60 ; each R 80 is independently R 70 or alternatively, two R 80 s, taken together with the nitrogen atom to which they are bonded, form a 5-, 6- or 7-membered heterocycloalkyl which may optionally include from 1 to 4 of the same or different additional heteroatoms selected from the group consisting of O, N and S, of which N may have -H or C-
  • Each M + may independently be, for example, an alkali ion, such as K + , Na + , Li + ; an ammonium ion, such as + N(R 60 ) 4 ; or an alkaline earth ion, such as [Ca 2+ ] 0 5 , [Mg 2+ ]o.5, or [Ba 2+ ]o.5
  • subscript 0.5 means that one of the counter ions for such divalent alkali earth ions can be an ionized form of a compound of the invention and the other a typical counter ion such as chloride, or two ionized compounds disclosed herein can serve as counter ions for such divalent alkali earth ions, or a doubly ionized compound of the invention can serve as the counter ion for such divalent alkali earth ions).
  • -NR R is meant to include -NH 2 , -NH-alkyl, /V-pyrrolidinyl, /V-piperazinyl, 4/V-methyl-piperazin-1-yl and N- morpholinyl.
  • substituent groups for hydrogens on unsaturated carbon atoms in "substituted" alkene, alkyne, aryl and heteroaryl groups are, unless otherwise specified, -R 60 , halo, -0 " M + , -OR 70 , -SR 70 , -S " M + , -NR 80 R 80 ,
  • R 60 , R 70 , R 80 and M + are as previously defined, provided that in case of substituted alkene or alkyne, the substituents are not -0 " M + , -OR 70 , -SR 70 , or -S " M + .
  • substituent groups for hydrogens on nitrogen atoms in "substituted" heteroalkyi and cycloheteroalkyi groups are, unless otherwise specified, -R 60 , -0 " M + , -OR 70 , -SR 70 , -S " M + , -NR 80 R 80 ,
  • a group that is substituted has 1 , 2, 3, or 4 substituents, 1 , 2, or 3 substituents, 1 or 2 substituents, or 1 substituent.
  • arylalkyloxycarbonyl refers to the group (aryl)-(alkyl)-0-C(0)-.
  • any of the groups disclosed herein which contain one or more substituents it is understood, of course, that such groups do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible.
  • the subject compounds include all stereochemical isomers arising from the substitution of these
  • pharmaceutically acceptable salt means a salt which is acceptable for administration to a patient, such as a mammal (salts with counterions having acceptable mammalian safety for a given dosage regime). Such salts can be derived from
  • “Pharmaceutically acceptable salt” refers to pharmaceutically acceptable salts of a compound, which salts are derived from a variety of organic and inorganic counter ions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, formate, tartrate, besylate, mesylate, acetate, maleate, oxalate, and the like.
  • salt thereof means a compound formed when a proton of an acid is replaced by a cation, such as a metal cation or an organic cation and the like.
  • the salt is a pharmaceutically acceptable salt, although this is not required for salts of intermediate compounds that are not intended for administration to a patient.
  • salts of the present compounds include those wherein the compound is protonated by an inorganic or organic acid to form a cation, with the conjugate base of the inorganic or organic acid as the anionic component of the salt.
  • solvent refers to a complex formed by combination of solvent molecules with molecules or ions of the solute.
  • the solvent can be an organic compound, an inorganic compound, or a mixture of both.
  • Some examples of solvents include, but are not limited to, methanol, /V,/V-dimethylformamide, tetrahydrofuran, dimethylsulfoxide, and water. When the solvent is water, the solvate formed is a hydrate.
  • Stereoisomers refer to compounds that have same atomic connectivity but different atomic arrangement in space. Stereoisomers include cis-trans isomers, H and Z isomers, enantiomers, and diastereomers.
  • pyrazoles imidazoles, benzimidazoles, triazoles, and tetrazoles.
  • “Pharmaceutically effective amount” and “therapeutically effective amount” refer to an amount of a compound sufficient to treat a specified disorder or disease or one or more of its symptoms and/or to prevent the occurrence of the disease or disorder.
  • a pharmaceutically or therapeutically effective amount comprises an amount sufficient to, among other things, cause the tumor to shrink or decrease the growth rate of the tumor.
  • mammal for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, cats, cows, sheep, goats, pigs, etc. In some cases, the mammal is human.
  • a target cell is an "inflicted” cell (e.g., a cell from an "inflicted” individual), where the term “inflicted” is used herein to refer to a subject with symptoms, an illness, or a disease that can be treated with a subject compound.
  • An "inflicted” subject can have cancer (e.g., HCC) and/or can have other hyper-proliferative conditions, for example sclerosis, fibrosis, and the like, etc.
  • “Inflicted cells” can be those cells that cause the symptoms, illness, or disease.
  • an inflicted cell of an inflicted patient can be a cancer cell (e.g., an hepatocellular carcinoma cell).
  • treatment used herein to generally refer to obtaining a desired pharmacologic and/or physiologic effect.
  • the effect can be prophylactic in terms of completely or partially preventing a disease or symptom(s) thereof and/or may be therapeutic in terms of a partial or complete stabilization or cure for a disease and/or adverse effect attributable to the disease.
  • treatment encompasses any treatment of a disease in a mammal, particularly a human, and includes: (a) preventing the disease and/or symptom(s) from occurring in a subject who may be predisposed to the disease or symptom but has not yet been diagnosed as having it; (b) inhibiting the disease and/or symptom(s), i.e., arresting their development; or (c) relieving the disease symptom(s), i.e., causing regression of the disease and/or symptom(s).
  • Those in need of treatment include those already inflicted (e.g., those with cancer) as well as those in which prevention is desired (e.g., those with increased susceptibility to cancer).
  • a therapeutic treatment is one in which the subject is inflicted prior to administration and a prophylactic treatment is one in which the subject is not inflicted prior to administration.
  • the subject has an increased likelihood of becoming inflicted or is suspected of being inflicted prior to treatment.
  • the subject is suspected of having an increased likelihood of becoming inflicted.
  • co-administration include the administration of two or more therapeutic agents either simultaneously, concurrently or sequentially within no specific time limits.
  • the agents are present in the cell or in the subject's body at the same time or exert their biological or therapeutic effect at the same time.
  • the therapeutic agents are in the same composition or unit dosage form. In other embodiments, the therapeutic agents are in separate compositions or unit dosage forms.
  • a first agent can be administered prior to (e.g., minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantly with, or subsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of a second therapeutic agent.
  • cancer includes any form of cancer, including but not limited to solid tumor cancers (e.g., lung, prostate, breast, bladder, colon, ovarian, pancreas, kidney, liver, glioblastoma, medulloblastoma, leiomyosarcoma, head & neck squamous cell carcinomas, melanomas, neuroendocrine; etc.) and liquid cancers (e.g., hematological cancers);
  • solid tumor cancers e.g., lung, prostate, breast, bladder, colon, ovarian, pancreas, kidney, liver, glioblastoma, medulloblastoma, leiomyosarcoma, head & neck squamous cell carcinomas, melanomas, neuroendocrine; etc.
  • liquid cancers e.g., hematological cancers
  • carcinomas soft tissue tumors; sarcomas; teratomas; melanomas; leukemias; lymphomas; and brain cancers, including minimal residual disease, and including both primary and metastatic tumors.
  • Carcinomas are malignancies that originate in the epithelial tissues. Epithelial cells cover the external surface of the body, line the internal cavities, and form the lining of glandular tissues. Examples of carcinomas include, but are not limited to: adenocarcinoma (cancer that begins in glandular (secretory) cells), e.g., cancers of the breast, pancreas, lung, prostate, and colon can be adenocarcinomas; adrenocortical carcinoma; hepatocellular carcinoma (HCC, malignant hepatoma); renal cell carcinoma; ovarian carcinoma; carcinoma in situ; ductal carcinoma; carcinoma of the breast; basal cell carcinoma; squamous cell carcinoma; transitional cell carcinoma; colon carcinoma; nasopharyngeal carcinoma; multilocular cystic renal cell carcinoma; oat cell carcinoma; large cell lung carcinoma; small cell lung carcinoma; non-small cell lung carcinoma; and the like. Carcinomas may be found in prostrate, pancreas, colon, brain (usually as secondary
  • Soft tissue tumors are a highly diverse group of rare tumors that are derived from connective tissue.
  • Examples of soft tissue tumors include, but are not limited to: alveolar soft part sarcoma; angiomatoid fibrous histiocytoma; chondromyoxid fibroma; skeletal tissue
  • chondrosarcoma extraskeletal myxoid chondrosarcoma; clear cell sarcoma; desmoplastic small round-cell tumor; dermatofibrosarcoma protuberans; endometrial stromal tumor; Ewing's sarcoma; fibromatosis (Desmoid); fibrosarcoma, infantile; gastrointestinal stromal tumor; bone giant cell tumor; tenosynovial giant cell tumor; inflammatory myofibroblastic tumor; uterine leiomyoma; leiomyosarcoma; lipoblastoma; typical lipoma; spindle cell or pleomorphic lipoma; atypical lipoma; chondroid lipoma; well-differentiated liposarcoma; myxoid/round cell
  • liposarcoma pleomorphic liposarcoma; myxoid malignant fibrous histiocytoma; high-grade malignant fibrous histiocytoma; myxofibrosarcoma; malignant peripheral nerve sheath tumor; mesothelioma; neuroblastoma; osteochondroma; osteosarcoma; primitive neuroectodermal tumor; alveolar rhabdomyosarcoma; embryonal rhabdomyosarcoma; benign or malignant schwannoma; synovial sarcoma; Evan's tumor; nodular fasciitis; desmoid-type fibromatosis; solitary fibrous tumor; dermatofibrosarcoma protuberans (DFSP); angiosarcoma; epithelioid hemangioendothelioma; tenosynovial giant cell tumor (TGCT); pigmented villonodular synovitis (PVNS);
  • a sarcoma is a rare type of cancer that arises in cells of mesenchymal origin, e.g., in bone or in the soft tissues of the body, including cartilage, fat, muscle, blood vessels, fibrous tissue, or other connective or supportive tissue.
  • Different types of sarcoma are based on where the cancer forms. For example, osteosarcoma forms in bone, liposarcoma forms in fat, and rhabdomyosarcoma forms in muscle.
  • Examples of sarcomas include, but are not limited to: askin's tumor; sarcoma botryoides; chondrosarcoma; ewing's sarcoma; malignant
  • hemangioendothelioma malignant schwannoma; osteosarcoma; and soft tissue sarcomas
  • soft tissue sarcomas e.g., alveolar soft part sarcoma; angiosarcoma; cystosarcoma phyllodesdermatofibrosarcoma protuberans (DFSP); desmoid tumor; desmoplastic small round cell tumor; epithelioid sarcoma; extraskeletal chondrosarcoma; extraskeletal osteosarcoma; fibrosarcoma; gastrointestinal stromal tumor (GIST); hemangiopericytoma; hemangiosarcoma (more commonly referred to as "angiosarcoma”); kaposi's sarcoma; leiomyosarcoma; liposarcoma; lymphangiosarcoma;
  • DFSP cystosarcoma phyllodesdermatofibrosarcoma protuberans
  • MPNST malignant peripheral nerve sheath tumor
  • neurofibrosarcoma neurofibrosarcoma
  • synovial sarcoma undifferentiated pleomorphic sarcoma, and the like.
  • a teratomas is a type of germ cell tumor that may contain several different types of tissue (e.g., can include tissues derived from any and/or all of the three germ layers: endoderm, mesoderm, and ectoderm), including for example, hair, muscle, and bone. Teratomas occur most often in the ovaries in women, the testicles in men, and the tailbone in children.
  • Melanoma is a form of cancer that begins in melanocytes (cells that make the pigment melanin). It may begin in a mole (skin melanoma), but can also begin in other pigmented tissues, such as in the eye or in the intestines.
  • Leukemias are cancers that start in blood-forming tissue, such as the bone marrow, and causes large numbers of abnormal blood cells to be produced and enter the bloodstream.
  • leukemias can originate in bone marrow-derived cells that normally mature in the bloodstream.
  • Leukemias are named for how quickly the disease develops and progresses (e.g., acute versus chronic) and for the type of white blood cell that is effected (e.g., myeloid versus lymphoid).
  • Myeloid leukemias are also called myelogenous or myeloblasts leukemias.
  • Lymphoid leukemias are also called lymphoblastic or lymphocytic leukemia. Lymphoid leukemia cells may collect in the lymph nodes, which can become swollen. Examples of leukemias include, but are not limited to: Acute myeloid leukemia (AML), Acute lymphoblastic leukemia (ALL), Chronic myeloid leukemia (CML), and Chronic lymphocytic leukemia (CLL).
  • AML Acute myeloid leukemia
  • ALL Acute lymphoblastic leukemia
  • CML Chronic myeloid leukemia
  • CLL Chronic lymphocytic leukemia
  • Lymphomas are cancers that begin in cells of the immune system.
  • lymphomas can originate in bone marrow-derived cells that normally mature in the lymphatic system.
  • One kind is Hodgkin lymphoma (HL), which is marked by the presence of a type of cell called the Reed-Stern berg cell.
  • HL Hodgkin lymphoma
  • Examples of Hodgkin lymphomas include: nodular sclerosis classical Hodgkin lymphoma (CHL), mixed cellularity CHL, lymphocyte-depletion CHL, lymphocyte-rich CHL, and nodular lymphocyte predominant HL.
  • NHL non-Hodgkin lymphomas
  • non-Hodgkin lymphomas include, but are not limited to: AIDS-related Lymphomas, anaplastic large-cell lymphoma, angioimmunoblastic lymphoma, blastic NK-cell lymphoma, Burkitt's lymphoma, Burkitt-like lymphoma (small non-cleaved cell lymphoma), chronic lymphocytic leukemia/small lymphocytic lymphoma, cutaneous T-Cell lymphoma, diffuse large B-Cell lymphoma, enteropathy-type T-Cell lymphoma, follicular lymphoma, hepatosplenic gamma-delta T-Cell lymphomas, T-Cell leukemias, lymphoblastic lymphoma, mantle cell lymphoma, marginal zone lymphoma, nasal T-Cell lymphoma, pediatric lymphoma, peripheral T-Cell lymphomas, primary central nervous system lymphoma, transformed lymphomas
  • Brain cancers include any cancer of the brain tissues.
  • Examples of brain cancers include, but are not limited to: gliomas (e.g., glioblastomas, astrocytomas, oligodendrogliomas, ependymomas, and the like), meningiomas, pituitary adenomas, vestibular schwannomas, primitive neuroectodermal tumors (medulloblastomas), etc.
  • embodiments are directed to racemic mixtures; pure stereoisomers (i.e., enantiomers or diastereomers); stereoisomer-enriched mixtures and the like unless otherwise indicated.
  • pure stereoisomers i.e., enantiomers or diastereomers
  • stereoisomer-enriched mixtures and the like unless otherwise indicated.
  • compositions of the present disclosure include compounds 1-7, shown below.
  • compositions and methods of the present disclosure also contemplate compounds 1-7.
  • Embodiments of the present disclosure include a compound that is a derivative of (2R ! 4aS ! 6aR ! 6aS ! 14aS ! 14bR)-10-hydroxy-2 ! 4a ! 6a ! 6a ! 9,14a-hexamethyl-1 1 -oxo- 1 ,3,4,5,6, 13,14, 14b-octahydropicene-2-carboxylic acid (Celastrol):
  • Embodiments of the present disclosure include a compound of formula (I):
  • the compounds described also include isotopically labeled compounds where one or more atoms have an atomic mass different from the atomic mass conventionally found in nature.
  • isotopes that may be incorporated into the compounds disclosed herein include, but are not limited to, 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18 0, 17 0, etc.
  • the subject compounds may be enriched in one or more of these isotopes relative to the natural abundance of such isotope.
  • deuterium ( 2 H) has a natural abundance of about 0.015%. Accordingly, for approximately every 6,500 hydrogen atoms occurring in nature, there is one deuterium atom.
  • Specifically contemplated herein are compounds enriched in deuterium at one or more positions.
  • deuterium containing compounds of the disclosure have deuterium at one or more positions (as the case may be) in an abundance of greater than 0.015%.
  • compositions that include a pharmaceutically acceptable carrier and a therapeutically effective amount of Compound 1 , Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, and/or Compound 7; or a pharmaceutically acceptable salt or solvate or stereoisomer thereof.
  • a subject compound can be administered alone, as the sole active pharmaceutical agent, or in combination with one or more additional subject compounds (e.g., any of
  • Compounds 1-7) or in conjunction with other agents e.g., in combination with an agent that is a standard of care for a given cancer such as hepatocellular carcinoma, e.g., Sorafenib; Linifanib; sunitinib; an HGF/c-MET inhibitor; an mTOR inhibitor, e.g., an mTOR inhibitor; and FGFR inhibitor; brivanib; tivantinib; cabozantinib; a CTLA-4 blocking compound such as an antibody such as tremelimumab; a PD-1 inhibitor, e.g., BMS-936558; an inhibitor of the Raf/MEK/ERK signaling pathway; an inhibitor of the PI3K AKT pathway; an inhibitor of the EGFR pathway; temsirolimus; rigosertib; imatinib; dasatinib; nilotinib; and the like; or, for example, in
  • the therapeutic agents can be formulated as separate compositions that are administered simultaneously or at different times, or the therapeutic agents can be administered together as a single composition combining two or more therapeutic agents.
  • the pharmaceutical compositions disclosed herein containing a subject compound optionally include other therapeutic agents. Accordingly, certain embodiments are directed to such pharmaceutical compositions, where the composition further includes a therapeutically effective amount of an agent selected as is known to those of skill in the art.
  • the subject compounds can antagonize the interaction of HSP90 with CDC37.
  • the compounds are useful for antagonizing the interaction of HSP90 with CDC37.
  • the compounds are useful for treating a disease or disorder (e.g., cancer such as a carcinoma such as hepatocellular carcinoma) that is mediated through the interaction of HSP90 with CDC37 in an individual (e.g., an individual having cancer, e.g., hepatocellular carcinoma).
  • a disease or disorder e.g., cancer such as a carcinoma such as hepatocellular carcinoma
  • an individual e.g., an individual having cancer, e.g., hepatocellular carcinoma.
  • the present disclosure provides a method of antagonizing the interaction of HSP90 with CDC37 (e.g., in a cell, in a cancer cell, in a biological sample, in an individual, in a cell of an individual, in a cancer cell of an individual, and the like).
  • the present disclosure provides a method of treating an individual having a disease or disorder that is mediated through the interaction of HSP90 with CDC37 in the individual.
  • the present disclosure provides a method of treating an individual having a cell proliferative disorder (e.g., cancer).
  • the present disclosure also provides a method of treating in individual having cancer (e.g., hepatocellular carcinoma).
  • cancer includes any form of cancer, including but not limited to solid tumor cancers (e.g., lung, prostate, breast, bladder, colon, ovarian, pancreas, kidney, liver, glioblastoma, medulloblastoma, leiomyosarcoma, head & neck squamous cell carcinomas, melanomas, neuroendocrine; etc.) and liquid cancers (e.g., hematological cancers); carcinomas (e.g., hepatocellular carcinoma); soft tissue tumors; sarcomas; teratomas; melanomas;
  • solid tumor cancers e.g., lung, prostate, breast, bladder, colon, ovarian, pancreas, kidney, liver, glioblastoma, medulloblastoma, leiomyosarcoma, head & neck squamous cell carcinomas, melanomas, neuroendocrine; etc.
  • liquid cancers e.g., hematological cancer
  • leukemias include lymphomas; and brain cancers, including minimal residual disease, and including both primary and metastatic tumors.
  • the disclosure also provides for a method for using a subject compound (e.g., any of compounds 1-7), a salt or solvate or stereoisomer thereof, as a research tool for studying a biological system or samples, or for discovering new chemical compounds having antagonistic properties toward the interaction of HSP90 and CDC37.
  • a subject compound e.g., any of compounds 1-7), a salt or solvate or stereoisomer thereof, as a research tool for studying a biological system or samples, or for discovering new chemical compounds having antagonistic properties toward the interaction of HSP90 and CDC37.
  • the embodiments are also directed to a subject compound (Compounds 1-7) or a salt or solvate or stereoisomer thereof, for use in therapy or as a medicament.
  • the embodiments are directed to the use of a subject compound or a salt or solvate or stereoisomer thereof, for the manufacture of a medicament; for example, for the manufacture of a medicament for antagonizing the interaction of HSP90 and CDC37.
  • the embodiments are also directed to the use of a subject compound or a salt or solvate or stereoisomer thereof for the manufacture of a medicament for the treatment of a disease or disorder mediated or sustained through the interaction of HSP90 with CDC37.
  • embodiments are also directed to the use of a subject compound or a salt or solvate or stereoisomer thereof for the manufacture of a medicament for the treatment of cancer (e.g., a carcinoma such as HCC).
  • the embodiments of the present disclosure are also directed to the use of a subject compound or a salt or solvate or stereoisomer thereof for the manufacture of a medicament for the treatment of a cell proliferative disorder.
  • Compounds as described herein can be purified by any purification protocol known in the art, including chromatography, such as HPLC, preparative thin layer chromatography, flash column chromatography and ion exchange chromatography. Any suitable stationary phase can be used, including normal and reversed phases as well as ionic resins.
  • the subject compounds are purified via silica gel and/or alumina chromatography. See, e.g., Introduction to Modern Liquid Chromatography, 2nd Edition, ed. L. R. Snyder and J. J. Kirkland, John Wiley and Sons, 1979; and Thin Layer Chromatography, ed E. Stahl, Springer-Verlag, New York, 1969.
  • the subject compounds can be synthesized via a variety of different synthetic routes using commercially available starting materials and/or starting materials prepared by
  • the amide Celstrol derivatives and the ester Celstrol derivatives of the subject compounds can be synthesized by contacting (2R,4aS,6aR,6aS,14aS,14bR)-10- hydroxy-2,4a,6a,6a,9,14a-hexamethyl-1 1 -oxo-1 ,3,4,5,6,13, 14,14b-octahydropicene-2-carboxylic acid (Celastrol) with an appropriate amine (e.g., benzylamine (1- phenylmethanamine) for cel- D1 , phenethylamine (2-phenylethylamine) for cel-D2; diethylamine (ethanamine) for cel-D3; or tert-butylamine (2-propanamine) for cel-D4) or appropriate alkyl halide (e.g. 2-bromopropane for cel-D6, benzyl bromide for cel-D7).
  • an appropriate amine
  • the compounds react in a polar aprotic solvent or a polar protic solvent.
  • the compounds react in a polar aprotic solvent.
  • the compounds react in a polar protic solvent.
  • Suitable polar aprotic solvents can include dimethylformamide, tetrahydrofuran, dimethylsulfoxide, dioxane, and the like.
  • Suitable polar protic solvents can include alcohols (e.g., isopropanol, methanol, ethanol, etc.), formic acid, and the like.
  • the reaction is carried out in the presence of a base.
  • the base can be a poor nucleophile (e.g., Diisopropylethylamine).
  • the reaction is carried out in the presence of a coupling reagent.
  • the coupling reagent is (Benzotriazol-1 - yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP).
  • a method of synthesizing a subject compound includes contacting (2R,4aS,6aR,6aS,14aS,14bR)-10- hydroxy-2,4a,6a,6a,9,14a-hexamethyl-1 1-oxo-1 ,3,4,5,6,13,14,14b-octahydropicene-2-carboxylic acid (Celastrol) with benzylamine (1 -phenylmethanamine) (e.g. to synthesize Compound 1 ), phenethylamine (2-phenylethylamine)(e.g., to synthesize Compound 2), diethylamine
  • reaction can be run at room temperature or can be heated.
  • the compounds react in a polar aprotic solvent or a polar protic solvent.
  • the compounds react in a polar aprotic solvent.
  • the compounds react in a polar protic solvent.
  • Suitable polar aprotic solvents can include dimethylformamide, tetrahydrofuran, dimethylsulfoxide, dioxane, and the like.
  • Suitable polar protic solvents can include alcohols (e.g., isopropanol, methanol, ethanol, etc.), formic acid, and the like.
  • the reaction is carried out in the presence of sodium bicarbonate. The reaction can be run at room temperature or can be heated.
  • the method further includes separating isomers with a resolution technique. In certain embodiments, in the above methods, the method further includes separating isomers with chiral chromatography.
  • the above methods further include the step of forming a salt of a subject compound.
  • Embodiments are directed to the other processes described herein, and to the product prepared by any of the processes described herein.
  • deionized water can be added to the reaction mixture.
  • the mixture is extracted (e.g., by ethyl acetate).
  • the combined organic extracts can be dried (e..g, over MgS04), and evaporated, filtered, and concentrated (e.g., via a rotary evaporator), which in some cases will yield a dark red oil.
  • the product can be purified (e.g., by flash chromatography (ethyl acetate:hexanes); reversed phase (RP)-HPLC, e.g., using a C18 column with an acetonitrile-water gradient mobile phase) and lyophilized, which in some cases will result in a dark orange to red solid.
  • RP reversed phase
  • the disclosed compounds are useful for the inhibition of PKC activity and the treatment of a disease or disorder that is mediated through the activity of a PKC activity. Accordingly, pharmaceutical compositions comprising at least one disclosed compound are also described herein.
  • a pharmaceutical composition that includes a subject compound may be administered to a patient alone, or in combination with other supplementary active agents.
  • the pharmaceutical compositions may be manufactured using any of a variety of processes, including, but not limited to, conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, lyophilizing, and the like.
  • the pharmaceutical composition can take any of a variety of forms including, but not limited to, a sterile solution, suspension, emulsion, lyophilisate, tablet, pill, pellet, capsule, powder, syrup, elixir or any other dosage form suitable for administration.
  • a subject compound may be administered to a subject using any convenient means capable of resulting in the desired reduction in disease condition or symptom.
  • a subject compound can be incorporated into a variety of formulations for therapeutic administration. More particularly, a subject compound can be formulated into pharmaceutical compositions by combination with appropriate pharmaceutically acceptable carriers or diluents, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, aerosols, and the like.
  • compositions that include at least one of the subject compounds can be formulated for use in human or veterinary medicine. Particular formulations of a disclosed pharmaceutical composition may depend, for example, on the mode of administration and/or on the location of the subject to be treated. In some embodiments, formulations include a pharmaceutically acceptable carrier in addition to at least one active ingredient, such as a subject compound. In other embodiments, other medicinal or pharmaceutical agents, for example, with similar, related or complementary effects on the disease or condition being treated can also be included as active ingredients in a
  • parenteral formulations may include injectable fluids, such as, but not limited to, pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle.
  • injectable fluids such as, but not limited to, pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle.
  • non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate.
  • compositions to be administered can optionally contain minor amounts of non-toxic auxiliary substances (e.g., excipients), such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like; for example, sodium acetate or sorbitan monolaurate.
  • excipients include, nonionic solubilizers, such as cremophor, or proteins, such as human serum albumin or plasma preparations.
  • materials which can serve as pharmaceutically-acceptable carriers include: (1 ) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (1 1 ) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydrox
  • compositions may be formulated as a pharmaceutically acceptable salt of a disclosed compound.
  • Pharmaceutically acceptable salts are non-toxic salts of a free base form of a compound that possesses the desired pharmacological activity of the free base. These salts may be derived from inorganic or organic acids. Non-limiting examples of suitable inorganic acids are hydrochloric acid, nitric acid, hydrobromic acid, sulfuric acid, hydroiodic acid, and phosphoric acid.
  • Non-limiting examples of suitable organic acids are acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, methyl sulfonic acid, salicylic acid, formic acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid, asparagic acid, aspartic acid,
  • the pharmaceutically acceptable salt includes formic acid. In certain embodiments, the pharmaceutically acceptable salt includes trifluoroacetic acid. Other suitable pharmaceutically acceptable salts are found in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., 1985. A pharmaceutically acceptable salt may also serve to adjust the osmotic pressure of the composition.
  • a subject compound can be used alone or in combination with appropriate additives to make tablets, powders, granules or capsules, for example, with conventional additives, such as lactose, mannitol, corn starch or potato starch; with binders, such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrators, such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants, such as talc or magnesium stearate; and if desired, with diluents, buffering agents, moistening agents, preservatives and flavoring agents.
  • Such preparations can be used for oral administration.
  • a subject compound can be formulated into preparations for injection by dissolving, suspending or emulsifying the compound in an aqueous or nonaqueous solvent, such as vegetable or other similar oils, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol; and if desired, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives.
  • the preparation may also be emulsified or the active ingredient encapsulated in liposome vehicles.
  • Formulations suitable for injection can be administered by an intravitreal, intraocular, intramuscular, subcutaneous, sublingual, or other route of administration, e.g., injection into the gum tissue or other oral tissue. Such formulations are also suitable for topical administration.
  • a subject compound can be delivered by a continuous delivery system.
  • continuous delivery system is used interchangeably herein with “controlled delivery system” and encompasses continuous (e.g., controlled) delivery devices (e.g., pumps) in combination with catheters, injection devices, and the like, a wide variety of which are known in the art.
  • a subject compound can be utilized in aerosol formulation to be administered via inhalation.
  • a subject compound can be formulated into pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen and the like.
  • a subject compound can be made into suppositories by mixing with a variety of bases such as emulsifying bases or water-soluble bases.
  • bases such as emulsifying bases or water-soluble bases.
  • a subject compound can be administered rectally via a suppository.
  • the suppository can include vehicles such as cocoa butter, carbowaxes and polyethylene glycols, which melt at body temperature, yet are substantially solid at room temperature.
  • unit dosage form refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a subject compound calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier or vehicle.
  • the specifications for a subject compound depend on the particular compound employed and the effect to be achieved, and the pharmacodynamics associated with each compound in the host.
  • the dosage form of a disclosed pharmaceutical composition may be determined by the mode of administration chosen.
  • topical or oral dosage forms may be employed.
  • Topical preparations may include eye drops, ointments, sprays and the like.
  • Oral formulations may be liquid (e.g., syrups, solutions or suspensions), or solid (e.g., powders, pills, tablets, or capsules). Methods of preparing such dosage forms are known, or will be apparent, to those skilled in the art.
  • compositions that include a subject compound may be formulated in unit dosage form suitable for individual administration of precise dosages.
  • the amount of active ingredient administered may depend on the subject being treated, the severity of the affliction, and the manner of administration, and is known to those skilled in the art.
  • the formulation to be administered contains a quantity of the compounds disclosed herein in an amount effective to achieve the desired effect in the subject being treated.
  • Each therapeutic compound can independently be in any dosage form, such as those described herein, and can also be administered in various ways, as described herein.
  • the compounds may be formulated together, in a single dosage unit (that is, combined together in one form such as capsule, tablet, powder, or liquid, etc.) as a combination product.
  • an individual subject compound may be administered at the same time as another therapeutic compound or sequentially, in any order thereof.
  • the subject compounds can antagonize the interaction of HSP90 with CDC37.
  • the compounds are useful for antagonizing the interaction of HSP90 with CDC37.
  • the compounds are useful for treating a disease or disorder (e.g., cancer such as a carcinoma such as hepatocellular carcinoma) that is mediated through the interaction of HSP90 with CDC37 in an individual (e.g., an individual having cancer, e.g., hepatocellular carcinoma).
  • a disease or disorder e.g., cancer such as a carcinoma such as hepatocellular carcinoma
  • an individual e.g., an individual having cancer, e.g., hepatocellular carcinoma.
  • the present disclosure provides a method of antagonizing the interaction of HSP90 with CDC37 (e.g., in a cell, in a cancer cell, in a biological sample, in an individual, in a cell of an individual, in a cancer cell of an individual, and the like).
  • the present disclosure provides a method of treating an individual having a disease or disorder that is mediated through the interaction of HSP90 with CDC37 in the individual.
  • the present disclosure provides a method of treating an individual having a cell proliferative disorder (e.g., cancer).
  • the present disclosure also provides a method of treating in individual having cancer (e.g., hepatocellular carcinoma).
  • the route of administration of a subject compound may be selected according to a variety of factors including, but not limited to, the condition to be treated, the formulation and/or device used, the patient to be treated, and the like.
  • administration useful in the disclosed methods include but are not limited to oral and parenteral routes, such as intravenous (iv), intraperitoneal (ip), rectal, topical, ophthalmic, nasal, and transdermal. Formulations for these dosage forms are described herein.
  • An effective amount of a subject compound may depend, at least, on the particular method of use, the subject being treated, the severity of the affliction, and the manner of administration of the therapeutic composition.
  • a "therapeutically effective amount" of a composition is a quantity of a specified compound sufficient to achieve a desired effect in an individual (e.g., patient) being treated. For example, this may be the amount of a subject compound necessary to prevent, inhibit, reduce or relieve a disease or disorder that is mediated through the interaction of HSP90 and CDC37 (e.g., hepatocellular carcinoma).
  • a therapeutically effective amount of a compound is an amount sufficient to prevent, inhibit, reduce or relieve a disease or disorder that is mediated through the interaction of HSP90 and CDC37 in an individual without causing a substantial cytotoxic effect on host cells (e.g., noncancerous cells of the individual).
  • Therapeutically effective doses of a subject compound or pharmaceutical composition can be determined by one of skill in the art, with a goal of achieving local (e.g., tissue) concentrations that are at least as high as the IC 50 of an applicable compound disclosed herein.
  • a dosage range is from 0.1 to 200 mg/kg body weight orally in single or divided doses.
  • a dosage range is from 1.0 to 100 mg/kg body weight orally in single or divided doses, including from 1.0 to 50 mg/kg body weight, from 1 .0 to 25 mg/kg body weight, from 1 .0 to 10 mg/kg body weight, from 0.5 to 25 mg/kg body weight, from 0.5 to 10 mg/kg body weight, from 5.0 to 25 mg/kg body weight, and/or from 5.0 to 10 mg/kg body weight (assuming an average body weight of approximately 70 kg; values may be adjusted accordingly for persons weighing more or less than average).
  • the compositions are, for example, provided in the form of a tablet containing from about 10 to about 1000 mg of the active ingredient, such as 25 to 750 mg, or 50 to 500 mg, for example 75 mg, 100 mg, 200 mg, 250 mg, 400 mg, 500 mg, 600 mg, 750 mg, or 1000 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject being treated.
  • a tablet containing from 500 mg to 1000 mg active ingredient is administered once (e.g., a loading dose) followed by administration of 1/2 (i.e., half) dosage tablets (e.g., from 250 to 500 mg) each 6 to 24 hours for 3 days or more.
  • the specific dose level and frequency of dosage for any particular subject may be varied and may depend upon a variety of factors, including the activity of the subject compound, the metabolic stability and length of action of that compound, the age, body weight, general health, sex and diet of the subject, mode and time of administration, rate of excretion, drug
  • Embodiments of the present disclosure also include combinations of one or more disclosed compounds with one or more other agents or therapies useful in the treatment of a disease or disorder.
  • the disease or disorder is mediated through the interaction of HSP90 with CDC37.
  • the disease or disorder is cell proliferative disorder (e.g., cancer).
  • one or more disclosed compounds may be administered in combination with therapeutically effective doses of other medicinal and pharmaceutical agents, or in combination other non-medicinal therapies, such as hormone or radiation therapy.
  • administration in combination with refers to both concurrent and sequential administration of the active agents.
  • a subject method can include a step to determine whether the interaction of HSP90 with CDC37 is antagonized (i.e., whether the interaction is reduced).
  • Antagonizing the interaction of HSP90 with CDC37 can be measured by monitoring the ability of HSP90 to specifically bind to CDC37 in a cell.
  • cell lysates can be prepared from cells that are contacted with a test Compound and a control compound (or no compound at all).
  • Immunoprecipitation can be performed using an antibody against CDC37, and then one can assay for the presence of HSP90 (e.g., via Western blot, mass spectrometry, etc.). Alternatively, immunoprecipitation can be performed using an antibody against HSP90, and then one can assay for the presence of CDC37 (e.g., via Western blot, mass spectrometry, etc.). Any method for detecting whether HSP90 forms a complex with (i.e., interacts with) CDC37 can be used.
  • test compound When less of an interaction is detected in the presence of a test compound compared to a control (e.g., a vehicle only control, or a compound known not to antagonize the interaction), then the test compound can be said to antagonize the interaction between HSP90 and CDC37.
  • the compound reduces the interaction to 90% or less (e.g., 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, or 5% or less) compared to control.
  • the compound when immunoprecipitating with an antibody against CDC37 and detecting HSP90, if the sample from the cells contacted with the test compound contains 90% the amount of HSP90 detected in the control sample, the compound can be said antagonize the interaction of HSP90 with CDC37 (e.g., reduce the interaction to 90% compared to control).
  • the efficacy of a subject compound can be tested by monitoring the presence of cancer (e.g., monitoring the presence of cancer cells, tumors, etc.) in the individual (e.g., following
  • HSP90AB1 Protein 90kDa alpha, class B member 1
  • HSP90AB1 HSP90 alpha
  • HSP90 beta HSP903
  • Many antibodies used to detect HSP90 detect both proteins.
  • a series of proteins including HSP90, HSP70, HSP20-30, and ubiquitin are induced by insults such as temperature shock, chemicals and other environmental stress.
  • a major function of HSP90 and other heat shock proteins (HSPs) is to act as molecular chaperones.
  • HSP 90 functions as a homodimer.
  • the encoded protein aids in the proper folding of specific target proteins by use of an ATPase activity that is modulated by co-chaperones.
  • Cell division cycle 37 (CDC37) is highly similar to Cdc37, a cell division cycle control protein of the yeast Sacchromyces cerevisiae. This protein is a molecular co-chaperone with specific function in cell signal transduction. It has been shown to form complex with Hsp90 and a variety of protein kinases including CDK4, CDK6, SRC, RAF-1 , MOK, as well as elF2 alpha kinases. It is thought to play a critical role in directing Hsp90 to its target kinases. For example, CDC 37 is over-expressed in hepatocellular carcinoma (HCC) cells, where it functions with HSP90 to regulate the activity of protein kinases in multiple oncogenic signaling pathways that contribute towards hepatocarcinogenesis.
  • HCC hepatocellular carcinoma
  • an article of manufacture containing materials useful for the treatment of the disorders described above comprises a container and a label.
  • Suitable containers include, for example, bottles, vials, syringes, and test tubes.
  • the containers may be formed from a variety of materials such as glass or plastic.
  • the container holds a composition which is effective for treating the condition and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle).
  • An active agent in the composition can be any of the subject compounds (Compounds 1 -7).
  • the label on, or associated with, the container can indicate that the composition is used for treating the condition of choice.
  • the article of manufacture may further comprise a second container comprising a pharmaceutically-acceptable buffer, such as phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.
  • a pharmaceutically-acceptable buffer such as phosphate-buffered saline, Ringer's solution and dextrose solution.
  • It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.
  • HSP90 and CDC37 chaperone complex would achieve anti-tumor effects in HCC, by inducing degradation and inhibiting phosphorylation of their client proteins kinases.
  • the recently identified HSP90/CDC37 antagonist, celastrol was evaluated for its anti-tumor activity in HCC cell lines and patient-derived xenografts.
  • derivatives of celastrol were synthesized and compared regarding safety and anti-tumor activity profiles in HCC patient-derived xenografts, a clinically relevant model for evaluating the performance of these compounds.
  • Celastrol and pristimerin were purchased from Sigma-Aldrich (St. Louis, MO). Two other celastrol derivatives, cel-D2 and cel-D7, were synthesized using celastrol as the starting material, using chemicals purchased from Sigma-Aldrich. Synthetic schemes for cel-D2 and cel- D7 are shown in Figure 9, and are exemplary for the synthesis of the subject compounds (amide derivatives: compounds 1-4; and ester derivatives: compounds 5-7). The 1 H and 13 C NMR spectra were obtained using the Varian 300 MHz or 400 MHz magnetic resonance spectrometer. High resolution mass spectrometric (MS) analyses of the compounds were performed at the Mass Spectrometry Facility at Stanford University.
  • MS mass spectrometric
  • RP-HPLC Reversed phase (RP)-HPLC (Dionex HPLC System; Dionex Corporation, Sunnyvale, CA) using a C18 column (Phenomenax, 5 ⁇ , 4.6 x 250 mm or Dionex, 5 ⁇ , 21.2 x 250 mm) and an acetonitrile-water gradient mobile phase (flow of 1 or 12 mL/minutes) afforded cel-D2 as an orange solid (17.2 mg, 68.0% yield), with m/z 554.3621 (M+H).
  • RP Reversed phase
  • cel-D7 For cel-D7, celastrol (20.2 mg, 0.045 mmol) was dissolved in dimethylformamide (2 mL), followed by addition of sodium bicarbonate (21 .5 mg, 0.256 mmol) and benzyl bromide (8 ⁇ , 0.067 mmol). After stirring for 24 hours at room temperature, deionized water (15 mL) was added and the reaction mixture was extracted by ethyl acetate (3 X 15 mL). The combined organic extracts were dried over MgS04, filtered and concentrated via a rotary evaporator to yield a dark red oil. RP-HPLC purification on a C18 column with an acetonitrile-water gradient afforded cel-D7 as an orange solid (14.0 mg, 57.4% yield), with m/z 541 .3306 (M+H).
  • DMEM Modified Eagle's Medium
  • fetal bovine serum Invitrogen, Carlsbad, CA
  • penicillin 100 ⁇ g/ml penicillin
  • streptomycin 100 ⁇ g/ml streptomycin.
  • Cells were cultured at 37°C in a humidified atmosphere with 5% C0 2 .
  • the HepG2 and Hep3B cell lines were obtained from American Type Culture Collection (Manassas, VA) in 2008.
  • the Huh7 cell line was a gift from Dr. Mark Kay (Stanford University, CA) in 2003. All cell lines were last
  • Total protein was extracted from tissues or harvested cells using T-PER Tissue Protein Extraction Reagent (Pierce; Rockford, IL), and protein concentration was determined with BCA Protein Assay Kit (Pierce, Rockford, IL). Equal amounts of protein (10 ⁇ g) were electrophoresed on 4% to 12% polyacrylamide gels (Invitrogen; Carlsbad, CA), and transferred onto
  • polyvinylidene difluoride membranes blocked with 10% nonfat milk for at least 1 hour, and probed with primary antibodies against CDC37 (ab61773) from Abeam (Cambridge, MA);
  • HSP90 (SC-59577), B-Raf (SC-166), GAPDH (SC-365062) from Santa Cruz Biotechnology (Santa Cruz, CA); CDK4 (2906), EGFR (2232), MEK1/2 (9122), AKT (9272), A-Raf (4432), C- Raf (9422), p-A-Raf (s299, 4431 ), p-B-Raf (s445,2696), p-C-Raf (s289/296/301 , 9431 ), p-AKT (s473, 4058s), p-MEK1/2 (s217/221 , 9121 ) from Cell Signaling ( Danvers, MA).
  • the specific proteins were detected with HRP-conjugated secondary antibodies (Santa Cruz Biotechnology; Santa Cruz, CA) and SuperSignal West Pico or West Femto Maximum Sensitivity substrate from Pierce (Rockford, IL). Co-immunoprecipitation was performed using Protein A G agarose (SC-2003, Santa Cruz Biotechnology, Santa Cruz, CA) according to manufacturer's instructions, with anti-CDC37 antibody (ab61773, Abeam, Cambridge, MA) for pull-down, and anti-HSP90 antibody (SC-59577, Santa Cruz Biotechnology, Santa Cruz, CA) for immunoblotting.
  • Test compounds were added at desired final concentrations, and further incubated for 72 hours before cell viability was assessed using CellTiter-Glo Luminescent Cell Viability Assay (Promega; Madison, Wisconsin) as previously described.
  • IC 50 s concentration of drug that inhibits cell growth by 50%
  • Three independent experiments were done, each in triplicates. Apoptosis analysis
  • Terminal dUTP-mediated nick-end labeling (TUNEL) assays were performed according to the manufacturer's protocol. Briefly, HepG2 cells were seeded in 8-chamber BD tissue culture slides (BD Bioscience Labware, Bedmord, MA) at 10% confluency. Celastrol or its derivatives were added to the culture medium at final concentrations of 5 ⁇ . After 6 hours incubation, cells were washed twice with PBS, and then fixed in 4% paraformaldehyde for 25 minutes. Fixed cells were washed twice in PBS with 0.1 % Triton X- 100, and then incubated with TUNEL reaction mixture for 60 minutes at 37°C.
  • TUNEL Terminal dUTP-mediated nick-end labeling
  • HCC tissues were collected from HCC patients who had undergone liver resection as part of their treatment. This study was approved by the Institutional Review Board at Stanford University for the use of human subjects in medical research, and informed consent was obtained from each patient prior to liver resection. Animal studies were carried out in compliance with Federal and local institutional rules for the conduct of animal experiments.
  • HCC specimens were mechanically and enzymatically dissociated in HBSS containing 0.1 % collagenase, 0.01 % hyaluronidase and 0.002% deoxyribonuclease at 37°C to obtain single cell suspensions. Cells were then passed through a 70- ⁇ filter, centrifuged at 100 g for 10 minutes and resuspended in Freezing Medium (FBS containing 10% DMSO) for storage at -80°C overnight, and transferred to liquid nitrogen for long-term storage.
  • FBS Freezing Medium
  • Single cell suspensions were then transduced with self-inactivating lentivirus carrying an ubiquitin promoter driving a trifusion reporter gene, which harbors a bioluminescence (Luc2), a fluorescence (egfp), and a positron emission tomography reporter gene (ttk) at a multiplicity of infection of 5.
  • High titer lentiviral vectors were produced using a modified version of a previous protocol. Tumor cells were stained with Pacific BlueTM anti- mouse CD45, Pacific BlueTM anti-mouse H-2Kd, and Pacific BlueTM anti-mouse CD31 (BioLegend; San Diego, CA).
  • Stable expressors were isolated by sorting as eGFP positive and Pacific Blue negative cells performed on a BD FACSAria (Becton Dickinson; Franklin Lakes, NJ).
  • Tumor size was measured with digital calipers and tumor volume was calculated using the formula ⁇ /6 x larger diameter x [smaller diameter] 2 .
  • Liver and tumor tissues were fixed in formalin and embedded with paraffin.
  • the 6- ⁇ tissue sections were stained with hematoxylin and eosin (H&E) for evaluation of cell morphology and apoptosis as described above.
  • H&E hematoxylin and eosin
  • mice in each group were euthanized and their bodies and harvested organs were weighed.
  • blood was collected for whole blood complete blood counts (CBC) and plasma chemistry analysis at the Stanford Animal Diagnostic Laboratory.
  • CBC whole blood complete blood counts
  • HSP90/CDC37 antagonists are feasible anti-tumor agents in the treatment of HCC
  • their selective cytotoxicity against HCC cells HepG2, Huh7, and Hep3B
  • normal hepatocytes Hu81 14, Hu4175, and Hu8130, obtained from three donors with non-diseased liver.
  • Western blot analysis confirmed that only HCC cells express high levels of CDC37, whereas normal hepatocytes express undetectable levels of CDC37 (Fig. 2A).
  • HSP90/CDC37 client protein kinases that are known to be highly activated in HCC, including the Raf family proteins, AKT, MEK1/2, CDK4, and EGFR.
  • HSP90/CDC37 client protein kinases studied, compared to DMSO control (Fig. 3). These effects were dose-dependent, with 10 ⁇ concentration of each compound causing greater reductions in expression and phosphorylation levels for almost all protein kinases compared to treatment with 1 ⁇ of each compound.
  • EGFR was not detected in HepG2 cells, but showed decreased levels after treatment with all compounds in Huh7 cells (with dose-dependence seen only with pristimerin and cel-D7).
  • CDC37 levels did not change after treatment with any compound.
  • celastrol reduced tumor volumes by 2-5 fold; pristimerin by 5-7 fold; cel-D2 by 1.5-3.5 fold; and cel-D7 by 1.8-3.2 fold.
  • the in vivo data were consistent with the in vitro data, with celastrol and pristimerin showing greater anti-tumor activity than cel-D2 and cel-D7.
  • tumor mass was absent from one of five mice in the group with HCC-1 xenografts after celastrol treatment, and from one of five mice in all three groups with HCC-1 , HCC-2, or HCC-3 xenografts after pristimerin treatment.
  • WBC white blood cells
  • RBC red blood cells
  • HGB hemoglobin
  • ALP alkaline phosphatase
  • AST aspartate aminotransferase
  • ALT alanine aminotransferase
  • BUN blood urea nitrogen.
  • Data represent mean ⁇ SD. (* p ⁇ 0.05%, compared to saline group.)
  • Pristimerin in particular showed strongest effects on AKT, MEK1/2 and EGFR, whereas celastrol showed strongest effect on B- Raf.
  • CDC37 expression in these xenografts were maintained (compared to original HCC tumor levels), and expression levels were unaffected by treatment with all four compounds.
  • HSP90 and its co-chaperone CDC37 are key factors in the chaperone-kinome pathway that plays permissive roles in the oncogenesis of multiple types of tumors.
  • celastrol and three of its derivatives were successfully shown to disrupt HSP90 and CDC37 interaction in HCC cells; to inhibit the growth of secondary HCC cell lines in vitro; and to inhibit the growth of orthotopic HCC patient-derived xenografts in vivo.
  • HCC cells such as Raf family proteins, AKT, MEK1/2, CDK4, and EGFR.
  • HSP90/CDC37 antagonists are potentially broad spectrum inhibitors that would be beneficial for treating the heterogeneous subtypes of HCC.
  • Our results suggest that HSP90/CDC37 antagonists can effectively inhibit the above (and other) pathways simultaneously, offering the benefit of treating a larger percentage of HCC patients without the need to administer chemotherapeutic cocktails that would also increase side effects.
  • An additional benefit resulting from simultaneous inhibition of a wide array of oncogenic kinase pathway is the potential to help overcome drug resistance that is often associated with the activation of one or more pathways.
  • sorafenib targeting angiogenesis and Raf/MEK ERK signaling pathways
  • adaptive drug resistance to EGFR-targeted therapies have been associated with activation of PI3K AKT pathway
  • therapies targeting AKT/mTOR leads to Raf/MEK/ERK pathway activation through a PI3K-dependent feedback loop.
  • HSP90/CDC37 antagonists either by themselves or in combination therapy with other pathway-specific inhibitors may potentially circumvent the development of acquired drug resistance by inhibiting multiple pathways simultaneously, and at the same time increase drug sensitivity. This is particularly beneficial for HCC, which is highly resistant to currently used chemotherapeutic drugs, making successful treatment a clinical challenge.
  • celastrol and pristimerin have greater anti-tumor efficacy but also greater toxicity both in vitro and in vivo (observed as lower IC 50 to normal hepatocytes in vitro and greater weight loss in vivo); whereas cel-D2 and cel-D7 have slightly reduced anti-tumor efficacy, but also have reduced toxicity (observed as higher IC 50 to normal hepatocytes in vitro and absence of weight loss in vivo).
  • Celastrol treatment also caused enlarged spleen and elevated white blood cells; whereas pristimerin treatment caused significant liver impairment (elevated AST and ALT level).
  • celastrol and pristimerin showed greatest inhibitory effects on the client protein kinases, which may underlie their greater apoptotic effects to both malignant and normal hepatocytes.
  • the structure-activity- relationship (SAR) study suggests that modifications of the carboxylic acid group of celastrol do not drastically affect its ability to disrupt HSP90/CDC37 inhibition; however, derivatives with aromatic phenyl substituents (cel-D2 and cel-D7) appear to induce less marked apoptosis and may therefore be less toxic to normal cells compared to derivatives with non-aromatic alkyl groups.
  • the clinical translation of celastrol and pristimerin may be limited due to their toxicity.
  • orthotopic patient-derived xenografts provides a high predictive value of the response of HCC patients to celastrol and its derivatives.
  • the human origin of these xenografts more accurately reflect the response rates; in fact, responses to chemotherapy in patient-derived xenografts have been reported to resemble the response rates of monotherapy in clinical trials.
  • Orthotopic models have also been shown to be predictive of a patient's response to chemotherapy.
  • Our xenografts are established from HCC patients with over- expression of CDC37, which we confirmed to be maintained in the xenografts.
  • HSP90/CDC37 antagonists can be used for the treatment of HCC, which are typically resistant to standard chemotherapeutic agents. They are broad spectrum agents, being able to simultaneously disrupt multiple oncogenic pathways that are critical in development of the heterogeneous subtypes of HCC. Thus, HSP90/CDC37 antagonists will be effective as monotherapy or as combination therapy with other conventional agents.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

L'invention concerne des composés qui sont utiles en tant qu'inhibiteurs de HSP90/CDC37 et qui sont par conséquent utiles pour traiter une variété de maladies et troubles qui sont médiés ou entretenus par l'intermédiaire de activité de HSP90 et CDC37. Cette invention concerne en outre des compositions pharmaceutiques comprenant ces composés, des méthodes d'utilisation de ces composés dans le traitement de diverses maladies ou divers troubles, et des procédés de préparation de ces composés.
PCT/US2015/039415 2014-07-08 2015-07-07 Antagonistes de hsp90/cdc37 et méthodes d'utilisation correspondantes WO2016007535A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201462022008P 2014-07-08 2014-07-08
US62/022,008 2014-07-08

Publications (1)

Publication Number Publication Date
WO2016007535A1 true WO2016007535A1 (fr) 2016-01-14

Family

ID=55064781

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2015/039415 WO2016007535A1 (fr) 2014-07-08 2015-07-07 Antagonistes de hsp90/cdc37 et méthodes d'utilisation correspondantes

Country Status (1)

Country Link
WO (1) WO2016007535A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108026142A (zh) * 2016-07-04 2018-05-11 厦门大学 孤儿核受体Nur77 的配体及其用途
CN109678923A (zh) * 2019-01-04 2019-04-26 中国药科大学 雷公藤红素（异）阿魏酸酯类衍生物及其制备方法与用途
CN113135977A (zh) * 2021-04-23 2021-07-20 沈阳药科大学 雷公藤红素-1,2,3-三氮唑衍生物及其制备方法和应用
CN113150058A (zh) * 2021-04-23 2021-07-23 沈阳药科大学 一种雷公藤红素衍生物及其制备方法和应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5650167A (en) * 1995-11-16 1997-07-22 Dawa Incorporated Method and composition for treating hepatitis B
US20090054438A1 (en) * 2007-08-17 2009-02-26 Burnham Institute For Medical Research Compositions and methods for inhibiting growth and metastasis of melanoma
US20110263693A1 (en) * 2006-03-31 2011-10-27 Dana-Farber Cancer Institute, Inc. Celastrol, gedunin, and derivatives thereof as hsp90 inhibitors

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5650167A (en) * 1995-11-16 1997-07-22 Dawa Incorporated Method and composition for treating hepatitis B
US20110263693A1 (en) * 2006-03-31 2011-10-27 Dana-Farber Cancer Institute, Inc. Celastrol, gedunin, and derivatives thereof as hsp90 inhibitors
US20090054438A1 (en) * 2007-08-17 2009-02-26 Burnham Institute For Medical Research Compositions and methods for inhibiting growth and metastasis of melanoma

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ZHANG, T ET AL.: "Characterization of Celastrol to Inhibit Hsp90 and Cdc37 Interaction", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 284, no. 51, 2009, pages 35381 - 35388 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108026142A (zh) * 2016-07-04 2018-05-11 厦门大学 孤儿核受体Nur77 的配体及其用途
CN109678923A (zh) * 2019-01-04 2019-04-26 中国药科大学 雷公藤红素（异）阿魏酸酯类衍生物及其制备方法与用途
CN109678923B (zh) * 2019-01-04 2021-04-27 中国药科大学 雷公藤红素（异）阿魏酸酯类衍生物及其制备方法与用途
CN113135977A (zh) * 2021-04-23 2021-07-20 沈阳药科大学 雷公藤红素-1,2,3-三氮唑衍生物及其制备方法和应用
CN113150058A (zh) * 2021-04-23 2021-07-23 沈阳药科大学 一种雷公藤红素衍生物及其制备方法和应用
CN113135977B (zh) * 2021-04-23 2022-04-22 沈阳药科大学 雷公藤红素-1,2,3-三氮唑衍生物及其制备方法和应用

Similar Documents

Publication Publication Date Title
Avril et al. Endoplasmic reticulum stress signaling and chemotherapy resistance in solid cancers
Yang et al. The role of autophagy in cancer: therapeutic implications
Huynh et al. RAD001 (everolimus) inhibits tumour growth in xenograft models of human hepatocellular carcinoma
Lee et al. A novel C-terminal heat shock protein 90 inhibitor that overcomes STAT3-Wnt-β-catenin signaling-mediated drug resistance and adverse effects
Wei et al. Novel celastrol derivatives inhibit the growth of hepatocellular carcinoma patient-derived xenografts
WO2016007535A1 (fr) Antagonistes de hsp90/cdc37 et méthodes d'utilisation correspondantes
CN112638889A (zh) 用于治疗的尿石素a和其衍生物
Crettol et al. In vitro P-glycoprotein-mediated transport of (R)-,(S)-,(R, S)-methadone, LAAM and their main metabolites
US9629830B2 (en) EphA4 inhibitors as neuroprotective agents
US9795595B2 (en) Methods for treating cancer
US20200360380A1 (en) Cancers expressing ccr5 and methods of treatment of same
Yu et al. NSK-01105 inhibits proliferation and induces apoptosis of prostate cancer cells by blocking the Raf/MEK/ERK and PI3K/Akt/mTOR signal pathways
WO2014160947A1 (fr) Inhibiteurs de diméthylarginine diméthylaminohydrolase et leurs procédés d'utilisation
Frączek et al. An outline of main factors of drug resistance influencing cancer therapy
EP3057593B1 (fr) Traitement du cancer du pancréas
Chen et al. TPGS-1000 exhibits potent anticancer activity for hepatocellular carcinoma in vitro and in vivo
US20240343728A1 (en) Compounds that inhibit pi3k isoform alpha and methods for treating cancer
Schweizer et al. From bevacizumab to tasquinimod: angiogenesis as a therapeutic target in prostate cancer
US20230065158A1 (en) Oxabicycloheptanes for treatment of small cell lung cancer
KR20220025849A (ko) 미토콘드리아의 표적화 및 암 줄기 세포의 근절을 위한 카르보시아닌 화합물
WO2012012653A1 (fr) Polythérapie basée sur des inhibiteurs de mdm2 et efgr
WO2013123033A1 (fr) Inhibiteurs de diméthylarginine diméthylaminohydrolase et procédés d'utilisation associés
Ghahremanloo et al. Synergistic anticancer effects of liposomal doxorubicin and aprepitant combination therapy in breast cancer: Preclinical insights and therapeutic potential
EP3137079A1 (fr) Inhibiteurs de jmjd2c utilisés en tant qu'agents anticancéreux
GUO Mechanistic Study of Artesunate Inhibiting B-cell Lymphoma and Treating Metabolic Diseases by Inducing Integrated Stress Response Pathway

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: 15819293

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15819293

Country of ref document: EP

Kind code of ref document: A1

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载