+

WO1997009877A1 - INHIBITION DE L'ACTIVATION DU FACTEUR NF-λB PAR LE CURCUMIN (DIFERULOYLMETHANE) - Google Patents

INHIBITION DE L'ACTIVATION DU FACTEUR NF-λB PAR LE CURCUMIN (DIFERULOYLMETHANE) Download PDF

Info

Publication number
WO1997009877A1
WO1997009877A1 PCT/US1996/014725 US9614725W WO9709877A1 WO 1997009877 A1 WO1997009877 A1 WO 1997009877A1 US 9614725 W US9614725 W US 9614725W WO 9709877 A1 WO9709877 A1 WO 9709877A1
Authority
WO
WIPO (PCT)
Prior art keywords
curcumin
activation
tnf
animal
cells
Prior art date
Application number
PCT/US1996/014725
Other languages
English (en)
Inventor
Bharat B. Aggarwal
Original Assignee
Research Development Foundation
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 Research Development Foundation filed Critical Research Development Foundation
Priority to AU69773/96A priority Critical patent/AU6977396A/en
Publication of WO1997009877A1 publication Critical patent/WO1997009877A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/12—Ketones

Definitions

  • the present invention relates generally to the fields of immunology and protein chemistry. More specifically, the present invention relates to the use of curcumin to inhibit activation of the transcription factor, NF-iB.
  • NF-iB transcription factor
  • These transcription factors are dimeric complexes composed of different members of the Rel/NF- ⁇ B family of polypeptides. This family is distinguished by the presence of a Rel homology domain of about 300 amino acids that displays a 35 percent to 61 percent identity between various family members (for references see 1) .
  • NF-icB is a ubiquitous transcription factor, it plays a critical role in the cells of the immune system, where it controls the expression of various cytokines and the major histocompatibility complex genes.
  • the inappropriate regulation of NF- ⁇ B and its dependent genes have been associated with various pathological conditions including toxic/septic shock, graft vs host reaction, radiation damage, atherosclerosis, and cancer (1, 2) .
  • NF- ⁇ B is an important target for therapeutic intervention.
  • the NF-JCB proteins and other members of the Rel family reside in the cytoplasm in an inactive state but upon activation, they are translocated to the nucleus.
  • the nuclear translocation of Rel proteins is induced by many agents, including inflammatory cytokines (e.g., tumor necrosis factor (TNF) , lymphotoxin (LT) , and interleukin (IL)-l), mitogens, bacterial products, protein synthesis inhibitors, oxidative stress (H 2 0 2 ) , ultraviolet light, and phorbol esters (3, 4).
  • TNF tumor necrosis factor
  • LT lymphotoxin
  • IL interleukin
  • Curcumin (diferuloylmethane) has been shown to block many reactions in which NF- ⁇ B plays a major role.
  • This agent is a major active component of turmeric (Curcuma lojiga) and gives specific flavor and yellow color to curry.
  • the compound has been shown to display anticarcinogenic properties in animals as indicated by its ability to inhibit both tumor initiation induced by benz (a) pyrene and 7,12 dimethylbenz (a) anthracene (5-8) and tumor promotion induced by phorbol esters (9,10), which are known to activate NF-KB.
  • Curcumin has also been shown to inhibit type 1 human immunodeficiency virus long terminal repeat (HIV-LTR) directed gene expression and virus replication stimulated by TNF and phorbol ester (11) , which likewise require NF-KB activation.
  • HIV-LTR type 1 human immunodeficiency virus long terminal repeat
  • TNF and phorbol ester (11)
  • the anti-inflammatory and antioxidant properties of curcumin have been well documented (12-14) . How these inhibitory responses are modulated by curcumin is not understood.
  • the prior art is deficient in the lack of effective means of inhibiting the activation of the transcription factor, NF-KB.
  • the present invention fulfills this longstanding need and desire in the art.
  • NF-/B a ubiquitous transcription factor
  • NF-KB a transcription factor that binds DNA as a heterodimeric complex composed of members of the Rel/NF-icB family of polypeptides. Because of its intimate involvement in host defense against disease, this transcription factor is an important target for therapeutic intervention.
  • curcumin diiferuloylmethane
  • NF-#B Treatment of human myeloid ML-la cells with TNF rapidly activated NF-#B, which consists of p50 and p65 subunits, and this activation was inhibited by curcumin.
  • AP-1 binding factors were also found to be down-modulated by curcumin, whereas the Spl binding factor was unaffected.
  • curcumin also blocked phorbol ester- and hydrogen peroxide-mediated activation of NF-/cB.
  • curcumin TNF-dependent phosphorylation and degradation of I/B ⁇ was not observed in curcumin-treated cells; the translocation of p65 subunit to the nucleus was inhibited at the same time.
  • the mechanism of action of curcumin was found to be different from that of protein tyrosine phosphatase inhibitors.
  • the present invention indicates that curcumin inhibits NF-JCB activation pathway at a step before IcB ⁇ phosphorylation but after the convergence of various stimuli.
  • curcumin is a potent inhibitor of NF- «B activation induced by various agents.
  • the results also indicate that curcumin inhibits at a step in the signal transduction cascade of NF- ⁇ B activation that occurs before I ⁇ B ⁇ phosphorylation but after the point at which various signals transduced by different stimuli converge.
  • the present invention shows that curcumin should be able to modulate NF-*B-dependent pathological conditions.
  • a method of inhibiting the activation of the NF-icB transcription factor in an animal in need of such treatment comprising the step of administering to said animal a pharmacologically effective dose of curcumin.
  • a method of inhibiting the nuclear translocation of the p65 subunit of the NF- ⁇ B transcription factor in a cell or in an animal in need of such treatment comprising the step of administering to said animal a pharmacologically effective dose of curcumin.
  • a method of inhibiting the phosphorylation and degradation of I/B ⁇ protein in a cell or in an animal in need of such treatment comprising the step of administering to said animal a pharmacologically effective dose of curcumin.
  • Figure 1 shows the dose response and kinetics of inhibition of TNF-dependent NF-KB by curcumin.
  • Figure la ML-la cells (2 x 10 6 /ml) were pre-incubated at 37°C for 60 minutes with different concentrations (2 to 60 ⁇ M) of curcumin followed by 30 minutes incubation with 0.1 nM TNF.
  • Figure lb ML-la cells (2 x 10 6 /ml) were preincubated at 37°C with 20 ⁇ M curcumin for different times and then measured NF- ⁇ B activation at 37°C for 30 minutes either with or without 0.1 nM TNF.
  • Figure 2 shows the supershift analysis and specificity of the effect of curcumin on the NF-JB activation.
  • nuclear extracts NE were prepared from untreated or TNF (0.1 nM)-treated ML-la cells (2 x 10 6 /ml) , incubated for 30 minutes with antibodies and then assayed for NF-*B as described below.
  • nuclear extracts prepared from TNF pre-treated cells were incubated with different concentrations of curcumin for 15 minutes and then analyzed for NF-/B by EMSA.
  • Figure 3 shows the effect of Curcumin on PMA- and H 2 0 2 - mediated activation of NF- ⁇ B.
  • ML-la cells (2 x 10 /ml) were preincubated for 60 minutes at 37°C with curcumin (50 ⁇ M) followed by PMA (25 ng/ml) or H 2 0 2 (0.5 mM) or indicated combinations for 30 minutes and then tested for NF- ⁇ B activation as described below.
  • Figure 4 shows the effect of curcumin on AP-1 and Spl transcription factors.
  • Cells were treated with different concentrations of curcumin for 60 minutes at 37°C, and nuclear extracts were then prepared and used for EMSA of AP-1 and Spl transcription factors as described below.
  • Figure 5 shows the effect of DTT and DMP on the Curcumin and PAO-induced inhibition of NF-/B activation.
  • ML-la (2 x 10 6 ml) were incubated at 37°C for 60 minutes with DTT (100 ⁇ M) or DMP (100 ⁇ M) in the presence of Curcumin (50 ⁇ M) or PAO (2.4 ⁇ M) or indicated combinations followed by 30 minutes incubation with 0.1 nM TNF and then assayed for NF-/cB activation as described below.
  • Figure 6 shows the effect of curcumin on TNF-induced phosphorylation and degradation of I/cB ⁇ and on level of p65 in cytoplasm and nucleus.
  • ML-la (2 x 10 6 /ml) pretreated for 60 minutes at 37 * C) with or without curcumin (50 ⁇ M) were incubated for different times with TNF (0.1 nM) , and then assayed for I*cB ⁇ ( Figure 6A) and for p65 ( Figure 6B) in cytosolic fractions by western blot analysis as described below.
  • ML-la (2 x l0 6 /ml) pretreated (for 60 minutes at 37°C) with curcumin were incubated with TNF (0.1 nM) for 30 minutes.
  • TNF tumor necrosis factor
  • PTPase protein tyrosine phosphatase
  • PTK protein tyrosine kinase
  • PPase protein phosphatase
  • PAO phenylarsine oxide
  • DTT dithiothreitol
  • DMP 2, 3-dimercaptopropanol
  • FBS fetal bovine serum
  • HIV-LTR human immunodeficiency virus-1 long terminal repeat.
  • the present invention is directed to a method of inhibiting the activation of the NF-icB transcription factor in an animal in need of such treatment comprising the step of administering to said animal a pharmacologically effective dose of curcumin.
  • the animal is a human.
  • the human has a pathophysiological state selected from the group consisting of toxic/septic shock, graft vs host reaction, radiation damage, atherosclerosis, and cancer.
  • the curcumin may be given in any dose which suitably inhibits the activation of the NF-icB transcription factor, the curcumin is preferably administered in a dose of from about l mg/kg to about 100 mg/kg.
  • the present invention is also directed to a method of inhibiting the nuclear translocation of the p65 subunit of the NF-KB transcription factor in a cell in an animal in need of such treatment comprising the step of administering to said animal a pharmacologically effective dose of curcumin.
  • the animal is preferably a human.
  • the human has a pathophysiological state selected from the group consisting of toxic/septic shock, graft vs host reaction, radiation damage, atherosclerosis, AIDS, inflammation and cancer.
  • the curcumin may be given in any dose which suitably inhibits the activation of the NF- ⁇ B transcription factor, the curcumin is preferably administered in a dose of from about 1 mg/kg to about 100 mg/kg.
  • the present invention also provides a method of inhibiting the phosphorylation and degradation of IicB ⁇ protein in a cell or in an animal in need of such treatment comprising the step of administering to said animal a pharmacologically effective dose of curcumin.
  • the animal is a human.
  • a method of inhibiting the phosphorylation and degradation of IicB ⁇ protein in a cell or in an animal is relevant where the human has a pathophysiological state selected from the group consisting of toxic/septic shock, graft vs host reaction, radiation damage, atherosclerosis, and cancer.
  • the curcumin is administered in a dose of from about 1 mg/kg to about 100 mg/kg.
  • Penicillin, streptomycin, RPMI 1640 medium, and fetal calf serum were obtained from GIBCO (Grand Island, NY) .
  • Curcumin, glycine, NaCl, and bovine serum albumin were obtained from Sigma Chemical Co. (St. Louis, MO) , and phenylarsine oxide from Aldrich Chemicals (Milwaukee, WI) .
  • Antibody against I/cB ⁇ , cyclin Dl and NF- ⁇ B subunits p50 and p65 and double stranded oligonucleotides having AP-1 and SP1 consensus sequences were obtained from Santa Cruz Biotechnology (Santa Cruz, CA) .
  • EXAMPLE 2 Cell lines The cell line employed was ML-la, a human myelomonoblastic leukemia cell line provided by Dr. Ken Takeda of Showa University, Japan. Cells were routinely grown in RPMI 1640 medium supplemented with giutamine (2 mM) , gentamicin (50 mg/ml) , and fetal bovine serum (FBS) (10 percent) . The cells were seeded at a density of lx 10 5 cells/ml in T25 flasks (Falcon 3013, Becton Dickinson Labware, Lincoln Park, NJ) containing 10 ml of medium and grown at 37 * C in an atmosphere of 95 percent air and 5 percent C0 2 . Cell cultures were split every 3 or 4 days. Occasionally, cells were tested for mycoplasma contamination using the DNA-based assay kit purchased from Gen-Probe (San Diego, CA) .
  • ML-la cells (2 x IO 6 cells/ml) were treated separately with different concentrations of an activator at 37°C. Nuclear extracts were then prepared according to Schreiber et al. (15) . Briefly, 2 x IO 6 cells were washed with cold phosphate-buffered saline (PBS) and suspended in 0.4 ml of lysis buffer (10 mM HEPES pH 7.9, 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1 mM DTT, 0.5 mM PMSF, 2.0 mg/ml leupeptin, 2.0 mg/ml aprotinin, and 0.5 mg/ml benzamidine) .
  • PBS cold phosphate-buffered saline
  • the cells were allowed to swell on ice for 15 minutes, after which 12.5 ml of 10 percent NP-40 was added.
  • the tube was then vigorously mixed on a vortex machine for 10 seconds, and the homogenate was centrifuged for 30 seconds in a microfuge.
  • the nuclear pellet was resuspended in 25 ⁇ l ice-cold nuclear extraction buffer (20 mM HEPES pH 7.9, 0.4 M NaCl, 1 mM EDTA, 1 mM EGTA, 1 mM DTT, 1 mM PMSF, 2.0 mg/ml leupeptin, 2.0 mg/ml aprotinin, and 0.5 mg/ml benzamidine), and the tube was incubated on ice for 30 minutes with intermittent mixing.
  • the tube was then centrifuged for 5 minutes in a microfuge at 4'C, and the supernatant (nuclear extract) was either used immediately or stored at -70 * C for later use.
  • the protein content was measured by the method of Bradford (16) .
  • Electrophoretic mobility shift assays were performed by incubating 4 mg of nuclear extract (NE) , with 16 fmoles of 32 P end-labeled 45-mer double-stranded NF-/cB oligonucleotide from the HIV-LTR, 5'-TTGTTACAAGGGACTTTCCGCTG GGGAjCTTTCCAGGGAGGCGTGG- 3', (17) for 15 minutes at 37°C.
  • the incubation mixture included 2-3 mg of poly (dl-dC) in a binding buffer (25 mM HEPES pH 7.9, 0.5 mM EDTA, 0.5 mM DTT, 1 percent NP-40, 5 percent glycerol, and 50 mM NaCl) (18, 19).
  • the DNA-protein complex formed was separated from free oligonucleotide on 4.5 percent native polyacrylamide gel using buffer containing 50 mM Tris, 200 mM glycine pH 8.5, and 1 mM EDTA (20) , and then the gel was dried.
  • nuclear extracts prepared from TNF treated cells were incubated with the antibodies against either p50 or p65 subunits of NF- ⁇ B for 30 minutes at room temperature before the complex was analyzed by EMSA (21) .
  • Antibodies against cyclin Dl and preimmune serum were included as negative controls.
  • EMSAs for AP-1 and Spl were performed as described for NF-JCB using 32 P end-labeled double-stranded oligonucleotides. Specificity of binding was determined routinely by using an excess of unlabeled oligonucleotide for competition as described earlier (21) . Visualization and quantitation of radioactive bands was carried out by a phosphorimager (Molecular Dynamics, Sunnyvale, CA) using 'Image-quant' software. EXAMPLE 4
  • ML-la cells (2 x 10° cells/ml) were treated separately with different concentrations of an activator at 37°C. Nuclear extracts were then prepared according to Schreiber et al. (15). Briefly, 2 x 10° cells were washed with cold phosphate-buffered saline (PBS) and suspended in 0.4 ml of lysis buffer (10 mM HEPES pH 7.9, 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1 mM DTT, 0.5 mM PMSF, 2.0 mg/ml leupeptin, 2.0 mg/ml aprotinin, and 0.5 mg/ml benzamidine).
  • PBS cold phosphate-buffered saline
  • the cells were allowed to swell on ice for 15 minutes, after which 12.5 ml of 10 percent NP-40 was added.
  • the tube was then vigorously mixed on a vortex machine for 10 seconds, and the homogenate was centrifuged for 30 seconds in a microfuge.
  • the nuclear pellet was resuspended in 25 ⁇ l ice-cold nuclear extraction buffer (20 mM HEPES pH 7.9, 0.4 M NaCl, 1 mM EDTA, 1 mM EGTA, 1 mM DTT, 1 mM PMSF, 2.0 mg/ml leupeptin, 2.0 mg/ml aprotinin, and 0.5 mg/ml benzamidine), and the tube was incubated on ice for 30 minutes with intermittent mixing.
  • the tube was then centrifuged for 5 minutes in a microfuge at 4 * C, and the supernatant (nuclear extract) was either used immediately or stored at -70 * C for later use.
  • the protein content was measured by the method of Bradford (16) .
  • EXAMPLE S Western Blotting for I/cBa and p65 After the NF-icB activation reaction described above, postnuclear extracts were resolved on 10 percent SDS-polyacrylamide gels for IjcBa. To determine p65 levels, nuclear and post nuclear (cytoplasmic) extracts were resolved on 8 percent SDS-polyacrylamide gels. After the gels, the proteins were electrotransferred to nitrocellulose filters, probed with a rabbit polyclonal antibody against I/Ba or against p65, and detected by chemiluminescence (ECL-Amersham) (22) . The bands obtained were quantitated using Personal Densitometer Scan vl.30 using Image Quanta software version 3.3 (Molecular Dynamics, Sunnyvale, CA) .
  • the present invention demonstrated the effect of curcumin on the activation of transcription factor NF-#cB.
  • Human ML-la cells were used for these studies because their response to NF-KB activation by various stimuli has been well characterized (21-23 ) .
  • the time of incubation and the concentration of the drugs used in the studies had no effect on the cell viability (data not shown) .
  • Curcumin inhibits TNF-Dependent NF- ⁇ B activation ML-la cells were preincubated for 1 hour with different concentrations of curcumin followed by treatment with TNF (0.1 nM) for 30 minutes at 37 ⁇ C. They were then examined for NF- ⁇ B activation by electrophoretic mobility shift assay. The results in Fig. la indicate that 40-60 ⁇ M curcumin inhibited most of the TNF response. Curcumin by itself did not activate NF- ⁇ B. The kinetics of inhibition were shown next, incubating the cells with curcumin either for 60, 30 and 10 minutes prior to the addition of TNF, at the same time as the addition of TNF, or 10 minutes after the addition of TNF. The cells were treated with TNF for 30 minutes. TNF response was inhibited only when cells were pretreated with curcumin ( Figure lb) . Cotreat ent of cells with TNF and curcumin was not effective.
  • a high concentration of TNF (10 nM) can activate NF-KB within 5 minutes and this induction is higher in its intensity than that obtained with cells using 100-fold lower concentration of TNF for longer time (23) .
  • curcumin-pretreated cells were exposed to 10 nM TNF for various times (Fig. lc) .
  • the induction of NF- ⁇ B by 10 nM TNF was very high and occurred within 5 minutes.
  • Curcumin could completely inhibit the activation of NF- ⁇ B induced by 10 nM as efficiently as it did with 0.1 nM TNF.
  • curcumin is a very potent inhibitor of NF-KB activation.
  • Curcumin down-modulates AP-1 but not Spl transcription factors Whether curcumin specifically blocks the activation of NF-KB or also affects other transcription factors was investigated. Curcumin had no effect on the Spl transcription factor (Fig. 4); however, DNA binding of AP-1 transcription factors was found to be down-modulated. This result is in agreement with an earlier report which showed that curcumin not only inhibits the DNA-binding activity of c-Jun/AP-1 binding factors but also down-modulates the level of these factors (24) .
  • DTT and DMP can also reverse the inhibitory effect of phenylarsine oxide (PAO; a potent PTPase inhibitor) on NF-JCB activation (21) .
  • PAO phenylarsine oxide
  • ML-la cells were treated with curcumin in the presence and absence of either DTT or DMP and then examined for the activation of NF-/cB by TNF.
  • DTT and DMP did not reverse the inhibition caused by curcumin but completely reversed the PAO-mediated inhibition.
  • Curcumin inhibits TNF-dependent phosphorylation and degradation of I/cBa and translocation of P65 subunit of NF- ⁇ B to the nucleus: The translocation of NF- ⁇ B to the nucleus is preceded by the phosphorylation and proteolytic degradation of I*cB ⁇ (26) .
  • the inhibitory action of curcumin was due to its effect on IJCBC. degradation, the cytoplasmic levels of I/cB protein were examined by western blot analysis. I ⁇ was phosphorylated within 5 minutes of TNF treatment of ML-la cells and then disappeared within 15 minutes. However, curcumin abolished both the phosphorylation (as indicated by absence of the slow migrating band) and degradation of I ⁇ a induced by TNF (Fig. 6A) .
  • the level of p65 in the cytoplasm and in the nucleus was also measured. As expected upon TNF treatment, the level of p65 declined in the cytoplasm with a concurrent increase in the nucleus (Fig 6B and 6C) . The treatment of cells with curcumin abolished the TNF-dependent change in the nuclear and cytoplasmic p65 levels. These results show that curcumin inhibits the TNF-induced translocation of p65 to the nucleus and this is consistent with the inhibition of TNF-dependent degradation of I ⁇ B curcumin.
  • Curcumin is a pharmacologically safe compound with known anti-inflammatory, anticarcinogenic, and free radical scavenger properties (6, 10, 27-30). However, how curcumin carries out these functions is not very clear.
  • NF-icB plays a pivotal role in cells of the immune system because it is rapidly activated by wide variety of pathogenic signals and functions as a potent and pleiotropic transcriptional activator. Intervention in NF-KB activation may be beneficial in suppressing toxic/septic shock, graft-vs-host reactions, acute inflammatory reactions, HIV replication, acute phase response and radiation damage.
  • the present invention demonstrates that curcumin completely blocked the TNF-dependent activation of NF- ⁇ B.
  • the activation induced by various other agents including phorbol ester and H 2 0 2 was also inhibited by curcumin.
  • the effect of curcumin was not due to the chemical modification of NF- ⁇ B proteins (25, 31, 52).
  • the inhibition of NF-KB activation was accompanied by the inhibition of p65 translocation to the nucleus and of I ⁇ B degradation.
  • TNF-activated signals including acidic and neutral sphingomyelinase-generatedceramides, proteases, serine/threonine protein kinase (PK) , protein tyrosine kinase (PTK) , protein tyrosine phosphatase (PTPase) and superoxide radicals in the activation of NF- ⁇ B have been implicated (1, 21, 22, 32-35). Whether these signals are generated by TNF sequentially or independently of each other, however, is not understood.
  • Curcumin may also block NF- ⁇ B activation by inhibiting a protein kinase.
  • curcumin has been shown to inhibit both serine/threonine PK and PTK (44) .
  • the PK needed for the activation of NF- ⁇ B has not, however, been identified.
  • PMA is an activator of PkC
  • both TNF and H 2 0 2 have been shown to activate both PkC and PTK.
  • NF- ⁇ B activation by TNF and H 2 0 2 has been shown to be blocked by inhibitors of both PKC and PTK (50) .
  • a PTK has also been implicated in NF- ⁇ B activation by ultraviolet light, lipopolysaccharide, hypoxia, and v-src (37-40, 51) .
  • TNF-dependent activation of NF- ⁇ B is dependent on erbstatin-sensitive PTK (22) .
  • Studies of Schieven et al 1993 (43) showed that PTK inhibitors block gamma-irradiation-induced NF- ⁇ B activation, a stimulant thought to work through the immediate generation of ROI, which suggest that PTK activation may precede ROI generation.
  • curcumin not only inhibits the DNA binding of c-jun/AP-1 transcription factor but it also down-modulates c-jun level by preventing its transcription (24) . Curcumin did not inhibit the Spl transcription factor under the same conditions in which it inhibited NF- ⁇ B and AP-1 transcription factors. Curcumin has also been shown to inhibit TNF and phorbol ester-stimulated type 1 HIV-LTR-directed gene expression and virus replication (11) , and this may be mediated through the inhibition of NF- ⁇ B.
  • curcumin can also inhibit nitric oxide synthase (45-47) . These observations can be explained based on these results since the expression of this enzyme is NF- ⁇ B dependent. This is consistent with the observation that TPCK, a protease inhibitor that blocks NF- ⁇ B activation, also blocks the expression of nitric oxide synthase (48) . TPCK, however, may exert its effect by a different mechanism than curcumin does. It has been shown that TPCK chemically modifies NF- ⁇ B, thus altering its release from I ⁇ Ba (25) . Curcumin, however, does not chemically modify the DNA binding properties of NF- ⁇ B.
  • Another level of modification that could prevent formation of p50/p65 heterodimer is down-modulation of the cytoplasmic pool of p65 subunit of NF- ⁇ B.
  • the present invention shows that p65 was not down-modulated by curcumin but its translocation to the nucleus was inhibited, most likely through inhibition of degradation of I ⁇ B.
  • curcumin has a high potential for use in modulating expression of genes regulated by NF-KB.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

La présente invention concerne un procédé pour inhiber l'activation du facteur de transcription NF-λB chez un animal nécessitant un tel traitement, consistant à lui administrer une dose de curcumin suffisante pour avoir un effet pharmacologique. L'invention concerne également un procédé pour inhiber la translocation nucléaire de la sous-unité p65 du facteur de transcription NF-λB dans une cellule ou chez un animal nécessitant un tel traitement, consistant à administrer une dose de curcumin suffisante pour avoir un effet pharmacologique.
PCT/US1996/014725 1995-09-14 1996-09-13 INHIBITION DE L'ACTIVATION DU FACTEUR NF-λB PAR LE CURCUMIN (DIFERULOYLMETHANE) WO1997009877A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU69773/96A AU6977396A (en) 1995-09-14 1996-09-13 Curcumin (diferuloylmethane) inhibition of nf$g(k)b activation

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US379995P 1995-09-14 1995-09-14
US60/003,799 1995-09-14

Publications (1)

Publication Number Publication Date
WO1997009877A1 true WO1997009877A1 (fr) 1997-03-20

Family

ID=21707670

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/014725 WO1997009877A1 (fr) 1995-09-14 1996-09-13 INHIBITION DE L'ACTIVATION DU FACTEUR NF-λB PAR LE CURCUMIN (DIFERULOYLMETHANE)

Country Status (3)

Country Link
AU (1) AU6977396A (fr)
WO (1) WO1997009877A1 (fr)
ZA (1) ZA967730B (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999065512A1 (fr) * 1998-06-19 1999-12-23 Japan As Represented By Director-General Of Agency Of Industrial Science And Technology Compositions medicinales contenant des peptides apparentes au facteur de croissance des fibroblastes
WO2000061167A3 (fr) * 1999-04-09 2001-01-11 Osteoscreen Inc TRAITEMENT DU SYNDROME MYELOPROLIFERATIF AU MOYEN D'INHIBITEURS DE L'ACTIVITE DES PROTEASOMES ET DE NF-λB
DE10029770A1 (de) * 2000-06-16 2001-12-20 Transmit Technologietransfer Verwendung von Curcuminen zur Behandlung von soliden Tumoren,insbesondere von Hirntumoren
US20110306675A1 (en) * 2002-02-21 2011-12-15 Kuwada Scott K Compositions and methods for inhibiting nf-kb mediated tumorigenicity and adhesion-dependent survival of cancer cells
WO2012163589A1 (fr) * 2011-05-27 2012-12-06 Unilever Plc Composition de polyphénols destinée à la prévention ou au traitement de l'athérosclérose

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5401777A (en) * 1991-11-14 1995-03-28 Steigerwald Arzneimittelwerk Gmbh Use of preparations of curcuma plants

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5401777A (en) * 1991-11-14 1995-03-28 Steigerwald Arzneimittelwerk Gmbh Use of preparations of curcuma plants

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BIOCHEMICAL PHARMACOLOGY, 26 May 1995, Vol. 49, No. 11, M.M. CHAN, "Inhibition of Tumor Necrosis Factor by Curcumin, A Phytochemical", pages 1551-1556. *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999065512A1 (fr) * 1998-06-19 1999-12-23 Japan As Represented By Director-General Of Agency Of Industrial Science And Technology Compositions medicinales contenant des peptides apparentes au facteur de croissance des fibroblastes
US6624146B1 (en) 1998-06-19 2003-09-23 Agency Of Industrial Science And Technology Pharmaceutical compositions containing fibroblast growth factor-related peptides
WO2000061167A3 (fr) * 1999-04-09 2001-01-11 Osteoscreen Inc TRAITEMENT DU SYNDROME MYELOPROLIFERATIF AU MOYEN D'INHIBITEURS DE L'ACTIVITE DES PROTEASOMES ET DE NF-λB
US6492333B1 (en) 1999-04-09 2002-12-10 Osteoscreen, Inc. Treatment of myeloma bone disease with proteasomal and NF-κB activity inhibitors
DE10029770A1 (de) * 2000-06-16 2001-12-20 Transmit Technologietransfer Verwendung von Curcuminen zur Behandlung von soliden Tumoren,insbesondere von Hirntumoren
US20110306675A1 (en) * 2002-02-21 2011-12-15 Kuwada Scott K Compositions and methods for inhibiting nf-kb mediated tumorigenicity and adhesion-dependent survival of cancer cells
WO2012163589A1 (fr) * 2011-05-27 2012-12-06 Unilever Plc Composition de polyphénols destinée à la prévention ou au traitement de l'athérosclérose

Also Published As

Publication number Publication date
ZA967730B (en) 1998-03-13
AU6977396A (en) 1997-04-01

Similar Documents

Publication Publication Date Title
US5891924A (en) Curcumin (diferuloylmethane) inhibition of NFκB activation
Xagorari et al. Luteolin inhibits an endotoxin-stimulated phosphorylation cascade and proinflammatory cytokine production in macrophages
Lee et al. Astaxanthin inhibits nitric oxide production and inflammatory gene expression by suppressing IκB kinase-dependent NF-κB activation
Manna et al. Immunosuppressive leflunomide metabolite (A77 1726) blocks TNF-dependent nuclear factor-κB activation and gene expression
Imai et al. The periodontal pathogen Porphyromonas gingivalis induces the Epstein–Barr virus lytic switch transactivator ZEBRA by histone modification
Singh et al. Capsaicin (8-methyl-N-vanillyl-6-nonenamide) is a potent inhibitor of nuclear transcription factor-kappa B activation by diverse agents.
Lin et al. Novel anti-inflammatory actions of nobiletin, a citrus polymethoxy flavonoid, on human synovial fibroblasts and mouse macrophages
Chabane et al. Histone deacetylase inhibitors suppress interleukin-1β-induced nitric oxide and prostaglandin E2 production in human chondrocytes
Rusyn et al. WY-14 643 rapidly activates nuclear factor kappaB in Kupffer cells before hepatocytes.
CA2289119C (fr) Procede de traitement de l'inflammation ou de maladies inflammatoires a l'aide d'inhibiteurs de poly-adp ribose polymerase
Grandjean-Laquerriere et al. Relative contribution of NF-κB and AP-1 in the modulation by curcumin and pyrrolidine dithiocarbamate of the UVB-induced cytokine expression by keratinocytes
Manna et al. Suppression of tumor necrosis factor-activated nuclear transcription factor-κB, activator protein-1, c-Jun N-terminal kinase, and apoptosis by β-lapachone
US5981583A (en) Inhibition of nuclear transcription factor NF-κB by caffeic acid phenethyl ester (CAPE), derivatives of CAPE, capsaicin (8-methyl-N-vanillyl-6-nonenamide) and resiniferatoxin
Yamazaki et al. (+)-Syringaresinol-di-O-β-d-glucoside, a phenolic compound from Acanthopanax senticosus Harms, suppresses proinflammatory mediators in SW982 human synovial sarcoma cells by inhibiting activating protein-1 and/or nuclear factor-κB activities
Tse et al. Magnolol suppresses NF-κB activation and NF-κB regulated gene expression through inhibition of IkappaB kinase activation
Kim et al. Suppression by a sesquiterpene lactone from Carpesium divaricatum of inducible nitric oxide synthase by inhibiting nuclear factor-κB activation
Umansky et al. Induction of NO synthesis in macrophages by Newcastle disease virus is associated with activation of nuclear factor-κB
Rödel et al. Dose‐dependent biphasic induction and transcriptional activity of nuclear factor kappa B (NF‐κ B) in EA. hy. 926 endothelial cells after low‐dose X‐irradiation
Koppal et al. Cyclopentenone prostaglandin 15-deoxy-Δ12, 14-prostaglandin J2 acts as a general inhibitor of inflammatory responses in activated BV-2 microglial cells
Ma et al. Osthole inhibits osteoclasts formation and bone resorption by regulating NF‐κB signaling and NFATc1 activations stimulated by RANKL
Fiebich et al. The non-steroidal anti-inflammatory drug tepoxalin inhibits interleukin-6 and alphal-anti-chymotrypsin synthesis in astrocytes by preventing degradation of IκB-alpha
Van Neerven et al. Inflammatory chemokine release of astrocytes in vitro is reduced by all‐trans retinoic acid
Kastelic et al. Induction of rapid IL-β mRNA degradation in THP-1 cells mediated through the AU-Rich region in the 3′ UTR by a radicicol analogue
Yeo et al. Avenanthramides attenuate inflammation and atrophy in muscle cells
Balci et al. Gossypol induced apoptosis in the human promyelocytic leukemia cell line HL 60

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AM AT AU BB BG BR BY CA CH CN CZ DE DK EE ES FI GB GE HU IL IS JP KE KG KP KR KZ LK LR LT LU LV MD MG MN MW MX NO NZ PL PT RO RU SD SE SG SI SK TJ TM TT UA UG UZ VN

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): KE LS MW SD SZ UG AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA

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