Echinomycin, a small-molecule inhibitor of hypoxia-inducible factor-1 DNA-binding activity.
Open accessPublished version of a preprint
Abstract
The identification of small molecules that inhibit the sequence-specific binding of transcription factors to DNA is an attractive approach for regulation of gene expression. Hypoxia-inducible factor-1 (HIF-1) is a transcription factor that controls genes involved in glycolysis, angiogenesis, migration, and invasion, all of which are important for tumor progression and metastasis. To identify inhibitors of HIF-1 DNA-binding activity, we expressed truncated HIF-1alpha and HIF-1beta proteins containing the basic-helix-loop-helix and PAS domains. Expressed recombinant HIF-1alpha and HIF-1beta proteins induced a specific DNA-binding activity to a double-stranded oligonucleotide containing a canonical hypoxia-responsive element (HRE). One hundred twenty-eight compounds previously identified in a HIF-1-targeted cell-based high-throughput screen of the National Cancer Institute 140,000 small-molecule library were tested in a 96-well plate ELISA for inhibition of HIF-1 DNA-binding activity. One of the most potent compounds identified, echinomycin (NSC-13502), a small-molecule known to bind DNA in a sequence-specific fashion, was further investigated. Electrophoretic mobility shift assay experiments showed that NSC-13502 inhibited binding of HIF-1alpha and HIF-1beta proteins to a HRE sequence but not binding of the corresponding proteins to activator protein-1 (AP-1) or nuclear factor-kappaB (NF-kappaB) consensus sequences. Interestingly, chromatin immunoprecipitation experiments showed that NSC-13502 specifically inhibited binding of HIF-1 to the HRE sequence contained in the vascular endothelial growth factor (VEGF) promoter but not binding of AP-1 or NF-kappaB to promoter regions of corresponding target genes. Accordingly, NSC-13502 inhibited hypoxic induction of luciferase in U251-HRE cells and VEGF mRNA expression in U251 cells. Our results indicate that it is possible to identify small molecules that inhibit HIF-1 DNA binding to endogenous promoters.
AI snapshot
Written from the paper’s full text.
TL;DR
Echinomycin (NSC-13502) was identified from a HIF-1-targeted compound screen as a sequence-specific DNA-binding inhibitor that blocks HIF-1 binding to hypoxia-responsive elements and reduces HIF-1-dependent transcription in U251 cells. It appears to act by binding DNA rather than by lowering HIF-1α protein, with selectivity over AP-1 and NF-κB in the assays used.
Key findings
- Of 128 active compounds from an NCI 140,000-compound HIF-1 cell-based screen, 20 inhibited HIF-1 DNA-binding activity by more than 50% at 10 µM; 12 of those also inhibited AP-1 and/or NF-κB binding, leaving 8 with relative HIF-1 selectivity.
- NSC-13502 dose-dependently inhibited recombinant HIF-1α/HIF-1β DNA binding in ELISA, with nearly complete inhibition at 320 nM, and in EMSA it did not inhibit AP-1 or NF-κB binding up to 10 µM.
- In ChIP assays, 0.5 µM NSC-13502 abolished HIF-1 binding to the VEGF promoter HRE but did not affect c-Fos binding to the cyclin D1 promoter or p65 binding to the ICAM-1 promoter; HIF-1α protein levels were not reduced.
- NSC-13502 inhibited hypoxic luciferase induction in U251-HRE cells with an EC50 of about 1.2 nM, blocked the roughly 5-fold hypoxic increase in VEGF mRNA (P < 0.01), and in microarray profiling 14 of 17 hypoxia-induced HIF-1 target genes were inhibited more than 3-fold.
Methods
- Recombinant truncated HIF-1α bHLH-PAS (amino acids 1–389) and HIF-1β bHLH-PAS (amino acids 1–475) proteins were expressed and used in a 96-well DNA-binding ELISA and EMSA with HRE, mutant HRE, AP-1, NF-κB, and E-box oligonucleotides; 128 compounds were screened at 10 µM.
- U251, U251-HRE, U251-pGL3, and MCF-7 cells were cultured under normoxia or 1% O2 hypoxia; assays included ChIP for VEGF, cyclin D1, and ICAM-1 promoters, Western blotting, luciferase reporter measurement, real-time PCR for VEGF, DNA thermal melting and ethidium bromide displacement, SRB viability, and oligonucleotide microarray analysis of about 11,000 genes.
- Main analyses compared dose-dependent inhibition, competition and supershift controls, promoter binding specificity, reporter activity, mRNA expression, and global gene-expression changes.
Limitations
- The paper states that NSC-13502’s sequence specificity may not be limited to HIF-1; it also inhibited nuclear protein binding to an E-box oligonucleotide, the binding site for Myc/Max.
- The paper states that in vivo feasibility and whether alternative dosing schedules could improve therapeutic potential remain to be established, and notes that earlier phase II clinical trials found minimal or no antitumor activity.
- Evident from the text: most mechanistic evidence comes from cell-free DNA-binding assays and cultured U251/MCF-7 cells, with no in vivo tumor efficacy experiment reported here; ChIP used 0.5 µM while the luciferase EC50 was about 1.2 nM, indicating different potency across assay contexts.
Model-written summary. Check it against the paper before relying on it.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
3 of 30 cited sources checked · 3 supported · 0 flagged
- Premisecitation
HIF-1α is overexpressed in human cancers due to hypoxia-dependent and/or hypoxia-independent pathways.
“Notably, HIF-1a is overexpressed in human cancers due to hypoxia-dependent and/or hypoxia-independent pathways (6,18,19)”
Quote in the paperSubstantial claim2 of 3 sources checked- cites [6] Karni R, Dor Y, Keshet E, Meyuhas O, Levitzki A. Activated pp60c-Src leads to elevated hypoxia-inducible factor (HIF)-1α expression under normoxia. J Biol Chem 2002; 277: 42919–25. source
- cites [18] Semenza G. Signal transduction to hypoxia-inducible factor 1. Biochem Pharmacol 2002; 64: 993–8. source
- cites [19] Laughner E, Taghavi P, Chiles K, Mahon PC, Semenza GL. HER2 (neu) signaling increases the rate of hypoxia-inducible factor 1α (HIF-1α) synthesis: novel mechanism for HIF-1-mediated vascular endothelial growth factor expression. Mol Cell Biol 2001; 21: 3995–4004. source
- Findingown data
NSC-13502 inhibits HIF-1 DNA-binding activity.
“We report here that echinomycin (NSC-13502) inhibits HIF-1 DNA-binding activity.”
- Also stated in: § Results · plain
“ELISA experiments (Fig. 2B) showed that NSC-13502 potently inhibited HIF-1a-389 and HIF-1h-475 DNA-binding activity in a dosedependent fashion, with almost complete inhibition at 320 nmol/L.”
- Also stated in: § Abstract · plain
“Electrophoretic mobility shift assay experiments showed that NSC-13502 inhibited binding of HIF-1A and HIF-1B proteins to a HRE sequence but not binding of the corresponding proteins to activator protein-1 (AP-1) or nuclear factor-KB (NF-KB) consensus sequences.”
Quote in the paperSubstantial claim - Also stated in: § Results · plain
- Findingown data
NSC-13502 does not affect HIF-1α mRNA expression in U251 cells.
“In contrast, HIF-1a mRNA expression, which was slightly decreased under hypoxic conditions relative to normoxia, was not affected by NSC-13502 (data not shown).”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 specifically inhibits HIF-1 binding to the HRE in the VEGF promoter in U251 cells treated with hypoxia and desferrioxamine.
“Treatment of U251 cells with NSC-13502 completely abrogated HIF-1 DNA-binding activity to the VEGF promoter.”
- Also stated in: § Abstract · plain
“Interestingly, chromatin immunoprecipitation experiments showed that NSC-13502 specifically inhibited binding of HIF-1 to the HRE sequence contained in the vascular endothelial growth factor (VEGF) promoter but not binding of AP-1 or NF-KB to promoter regions of corresponding target genes.”
Quote in the paperSubstantial claim - Also stated in: § Abstract · plain
- Findingown data
NSC-13502 does not inhibit AP-1 DNA-binding activity induced by c-Fos and JunD at concentrations up to 10 μmol/L.
“More importantly, NSC-13502 up to 10 Amol/L did not inhibit DNA-binding activities induced by c-Fos and JunD to the corres-ponding AP-1 DNA-binding sequence (Fig. 2C-b)”
Quote in the paperSubstantial claim
- Findingown data
NSC-13502 does not affect cyclin D1 protein levels in U251 cells under hypoxia.
“In addition, cyclin D1 protein levels were not affected by treatment under hypoxia or with NSC-13502, consistent with data obtained in the ChIP assay.”
Quote in the paperSubstantial claim - Findingown data
Treatment of hypoxic U251 cells with NSC-13502 has minimal effect on constitutive and inducible HIF-1 DNA-binding activities detected in nuclear extracts.
“Treatment of hypoxic U251 cells with NSC-13502 had a minimal effect on both constitutive and inducible HIF-1 DNA-binding activities detected in nuclear extracts (Fig. 2D-b, lane 3).”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 does not significantly affect luciferase expression in U251-pGL3 control cells under normoxic or hypoxic conditions.
“In contrast, NSC-13502 did not significantly affect luciferase expression in the U251-pGL3 control cell line under either normoxic or hypoxic conditions (Fig. 4B).”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 inhibits hypoxic induction of luciferase expression in U251-HRE cells in a dose-dependent fashion with an EC50 of approximately 1.2 nmol/L.
“As shown in Fig. 4A, NSC-13502 very potently inhibited hypoxic induction of luciferase expression in U251-HRE in a dose-dependent fashion with an EC 50 of f1.2 nmol/L.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 does not inhibit p65 binding to the NF-κB consensus sequence at concentrations up to 10 μmol/L.
“More importantly, NSC-13502 up to 10 Amol/L did not inhibit DNA-binding activities induced by c-Fos and JunD to the corres-ponding AP-1 DNA-binding sequence (Fig. 2C-b) or the binding of p65 to the NF-nB consensus sequence (Fig. 2C-c).”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 increases the melting temperature of HRE DNA from 51.5°C to 65°C.
“The melting temperature (T m ) of HRE DNA was increased from 51.5jC to 65jC (26.2% increase), whereas only a 5.5jC increase for NF-nB binding and a 5jC increase for AP-1-binding oligonucleotides were observed”
Quote in the paperSubstantial claim - Findingown data
Hypoxia induces a 5-fold increase in VEGF mRNA expression in U251 cells.
“As shown in Fig. 5A, hypoxia induced a 5-fold increase in VEGF mRNA expression, which was significantly (P < 0.01) inhibited by addition of NSC-13502 in a dose-dependent fashion.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 inhibits hypoxic induction of VEGF mRNA expression in U251 cells in a dose-dependent manner.
“As shown in Fig. 5A, hypoxia induced a 5-fold increase in VEGF mRNA expression, which was significantly (P < 0.01) inhibited by addition of NSC-13502 in a dose-dependent fashion.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 does not affect binding of NF-κB (p65) to the ICAM-1 promoter in U251 cells.
“Importantly, NSC-13502 did not affect the binding of AP-1 (c-Fos) to the cyclin D1 promoter or the binding of NF-nB (p65) to the ICAM-1 promoter, transcription factors that were constitutively present in untreated U251 cells.”
Quote in the paperSubstantial claim - Findingown data
Recombinant HIF-1α-389 and HIF-1β-475 proteins mediate specific DNA-binding activity to an oligonucleotide containing the canonical HIF-1-binding site (HRE).
“Expressed recombinant HIF-1A and HIF-1B proteins induced a specific DNA-binding activity to a double-stranded oligonucleotide containing a canonical hypoxia-responsive element (HRE).”
- Also stated in: § Results · plain
“EMSA experiments showed that HIF-1a-389 and HIF-1h-475 recombinant proteins mediated a specific DNA-binding activity to an oligonucleotide containing the canonical HIF-1-binding site (HRE), which was competed for by addition of excess unlabeled WT, but not mutant, oligonucleotide (Fig. 1B).”
Quote in the paperSubstantial claim - Also stated in: § Results · plain
- Findingown data
Addition of NSC-13502 to the binding reaction in vitro completely abrogates HIF-1-dependent DNA-binding activities detected in nuclear extracts from hypoxic U251 cells.
“In contrast, addition of NSC-13502 to the binding reaction in vitro completely abrogated HIF-1-dependent DNA-binding activities detected in nuclear extracts from hypoxic U251 cells (Fig. 2D-b, lane 4).”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 does not affect HIF-1α protein levels in U251 cells under hypoxia.
“In conclusion, NSC-13502 does not affect HIF-1a protein levels and does not have a general effect on other proteins.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 does not affect binding of AP-1 (c-Fos) to the cyclin D1 promoter in U251 cells.
“Importantly, NSC-13502 did not affect the binding of AP-1 (c-Fos) to the cyclin D1 promoter or the binding of NF-nB (p65) to the ICAM-1 promoter, transcription factors that were constitutively present in untreated U251 cells.”
Quote in the paperSubstantial claim - Findingown data
Addition of NSC-13502 to the DNA-binding reaction completely abrogated binding of nuclear proteins to the E-box in MCF-7 nuclear extracts.
“Interestingly, addition of NSC-13502 to the DNA-binding reaction completely abrogated binding of nuclear proteins to the DNA (lane 4), providing further evidence of the sequence specificity of NSC-13502.”
Quote in the paperSubstantial claim - Findingown data
14 of the 17 HIF-1-inducible genes were significantly inhibited by NSC-13502 by >3-fold.
“Fourteen of the 17 HIF-1-inducible genes were significantly inhibited by NSC-13502 by >3-fold (Table 1).”
Quote in the paperSubstantial claim - Findingown data
All 20 compounds inhibited HIF-1 DNA-binding activity in EMSA.
“EMSA results showed that all 20 compounds inhibited HIF-1 DNA-binding activity, consistent with results obtained in ELISA.”
Quote in the paperSubstantial claim - Findingown data
In EMSA, NSC-13502 showed a significant degree of specificity between inhibiting binding of HIF-1 to the HRE, which contains 5'-ACGT-3' in its core sequence, and unrelated sequences recognized by the AP-1 or NF-κB family of transcription factors.
“In EMSA, NSC-13502 showed a significant degree of specificity between inhibiting binding of HIF-1 to the HRE, which contains 5V-ACGT-3V in its core sequence, and unrelated sequences recognized by the AP-1 or NF-nB family of transcription factors.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 inhibition of luciferase expression in U251-HRE is reversible.
“As shown in Fig. 4C, 94%, 80%, and 42% of hypoxiainduced luciferase expression was restored in cells in which NSC-13502 was washed out after 4-hour treatment at 5, 10, and 20 nmol/L, respectively, indicating that the DNA-binding activity of NSC-13502 is in fact reversible.”
Quote in the paperNot a substantial claim - Findingown data
NSC-13502 inhibited hypoxic induction of luciferase in U251-HRE cells.
“Consistent with data from the primary high-throughput screen, NSC-13502 inhibited hypoxic induction of luciferase in U251-HRE cells”
- Also stated in: § Discussion · plain
“NSC-13502 inhibits hypoxic induction of luciferase expression in U251-HRE cells.”
Quote in the paperSubstantial claim - Also stated in: § Discussion · plain
- Findingown data
10 known HIF-1-inducible genes were identified among the 65 genes inhibited >4-fold by NSC-13502 under hypoxic conditions.
“Moreover, 10 known HIF-1-inducible genes were identified among the 65 genes inhibited >4-fold by NSC-13502 under hypoxic conditions.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 blocks HIF-1 DNA binding of endogenous nuclear proteins from hypoxic-treated cells only when the compound is added directly to the binding reaction.
“the ability of NSC-13502 to block HIF-1 DNA binding of endogenous nuclear proteins from hypoxic treated cells, but only when the compound was added directly to the binding reaction”
Quote in the paperSubstantial claim - Findingown data
Eight compounds showed relative specificity for inhibition of HIF-1 DNA binding.
“Eight compounds showed a relative specificity for inhibition of HIF-1 DNA binding (Supplementary Fig. S1).”
Quote in the paperSubstantial claim - Findingown data
More genes are down-regulated under hypoxic than normoxic conditions in U251 cells treated with NSC-13502 (451 vs 276 by ≥2-fold; 65 vs 36 by ≥4-fold).
“Notably, more genes were down-regulated under hypoxic than normoxic conditions (451 versus 276 by z2-fold and 65 versus 36 by z4-fold)”
Quote in the paperSubstantial claim - Findingown data
Hypoxia induces HIF-1α protein accumulation in U251 cells.
“As shown in Fig. 3B, hypoxia induced HIF-1a protein accumulation, which was slightly increased by addition of NSC-13502.”
Quote in the paperSubstantial claim - Findingown data
Twelve of 20 compounds also inhibited either AP-1 or NF-κB DNA binding or both.
“However, 12 of 20 compounds also inhibited either AP-1 or NF-nB DNA binding or both, suggesting that these compounds were nonspecific inhibitors of protein DNA binding (data not shown).”
Quote in the paperSubstantial claim - Findingcitation
17 of 109 genes induced by hypoxia by >2.5-fold could be identified as HIF-1-inducible genes based on published results.
“In particular, 17 of 109 genes induced by hypoxia by >2.5-fold could be identified as HIF-1-inducible genes based on published results.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 inhibited VEGF mRNA expression in U251 cells.
“NSC-13502 inhibited hypoxic induction of luciferase in U251-HRE cells and VEGF mRNA expression in U251 cells.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 slightly increases hypoxia-induced HIF-1α protein accumulation.
“As shown in Fig. 3B, hypoxia induced HIF-1a protein accumulation, which was slightly increased by addition of NSC-13502.”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 blocks HIF-1 DNA binding to the HRE of the VEGF promoter detected by ChIP.
“the ability of NSC-13502 to block HIF-1 DNA binding to the HRE of VEGF promoter detected by ChIP.”
- Also stated in: § Discussion · plain
“NSC-13502 inhibits HIF-1 binding to the HRE of the VEGF promoter in cultured cells.”
Quote in the paperSubstantial claim - Also stated in: § Discussion · plain
- Findingown data
More genes were down-regulated than up-regulated by NSC-13502 in U251 cells.
“Overall, we found that more genes were down-regulated than up-regulated”
Quote in the paperNot a substantial claim - Findingown data
Twenty of 128 compounds inhibited HIF-1 DNA-binding activity by >50% relative to untreated control at 10 μmol/L.
“Twenty of 128 compounds inhibited HIF-1h-389 and HIF-1h-475 DNA-binding activity by >50% relative to untreated control at the 10 Amol/L concentration (data not shown).”
Quote in the paperSubstantial claim - Findingown data
NSC-13502 completely inhibited DNA-binding activities to an oligonucleotide containing the E-box sequence detected in nuclear extracts harvested from untreated MCF-7 cells.
“Indeed, we found that NSC-13502 completely inhibited DNA-binding activities to an oligonucleotide containing the E-box sequence detected in nuclear extracts harvested from untreated MCF-7 cells.”
Quote in the paperSubstantial claim - Interpretationown data
NSC-13502 inhibits DNA binding of endogenous full-length HIF-1α and HIF-1β proteins.
“These results show that NSC-13502 inhibits DNA binding of endogenous full-length HIF-1a and HIF-1h proteins and are consistent with the possibility that NSC-13502 inhibits HIF-1 DNA binding by interacting with DNA and not by interfering with protein-protein interaction.”
Quote in the paperSubstantial claim - Interpretationown data
It is possible to identify small molecules that inhibit HIF-1 DNA binding to endogenous promoters.
“Our results indicate that it is possible to identify small molecules that inhibit HIF-1 DNA binding to endogenous promoters.”
- Also stated in: § Introduction · hedged
“Our data provide proof-of-principle that it is possible to identify small molecules that specifically inhibit HIF-1 DNA binding”
Quote in the paperSubstantial claim - Also stated in: § Introduction · hedged
- Premisecitation
Echinomycin (NSC-13502) is a small molecule known to bind DNA in a sequence-specific fashion.
“echinomycin (NSC-13502), a smallmolecule known to bind DNA in a sequence-specific fashion”
- Also stated in: § Results · boosted
“NSC-13502 is a small molecule known to bind DNA in a sequence-specific fashion (15).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [15] Van Dyke MM, Dervan PB. Echinomycin binding sites on DNA. Science 1984; 225: 1122–7. source
- Also stated in: § Results · boosted
- Interpretationown data
NSC-13502 exhibits a higher affinity for HRE core sequence than AP-1- or NF-κB-binding core sequences.
“indicating that NSC-13502 exhibits a higher affinity for HRE core sequence than AP-1-or NF-nB-binding core sequences.”
Quote in the paperSubstantial claim - Interpretationown data
NSC-13502 inhibited preferentially HIF-1-dependent gene expression.
“consistent with the possibility that NSC-13502 inhibited preferentially HIF-1-dependent gene expression.”
- Also stated in: § Results · hedged
“In conclusion, our results show that expression of a limited number of genes is affected by treatment of U251 cells with NSC-13502 and they are consistent with a predictable pattern of inhibition of known HIF-1-inducible genes.”
Quote in the paperSubstantial claim - Also stated in: § Results · hedged
- Premisecitation
HIF-1 induces expression of genes whose products are involved in cancer cell survival, glycolysis, angiogenesis, migration, and invasion.
“HIF-1 induces expression of genes whose products are involved in cancer cells survival, glycolysis, angiogenesis, migration, and invasion (5).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [5] Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer 2003; 3: 721–32. source
- Premisecitation
Minimal or no antitumor activity was found in phase II clinical trials of NSC-13502.
“However, minimal or no antitumor activity was found in phase II clinical trials.”
Quote in the paperSubstantial claim0 of 4 sources checked- cites [47] Foster BJ, Clagett-Carr K, Shoemaker DD, et al. Echinomycin: the first bifunctional intercalating agent in clinical trials. Invest New Drugs 1985; 3: 403–10. source
- cites [48] Muss HB, Blessing JA, DuBeshter B. Echinomycin in recurrent and metastatic endometrial carcinoma. A phase II trial of the Gynecologic Oncology Group. Am J Clin Oncol 1993; 16: 492–3. source
- cites [49] Wadler S, Tenteromano L, Cazenave L, et al. Phase II trial of echinomycin in patients with advanced or recurrent colorectal cancer. Cancer Chemother Pharmacol 1994; 34: 266–9. source
- cites [50] Chang AY, Kim K, Boucher H, et al. A randomized phase II trial of echinomycin, trimetrexate, and cisplatin plus etoposide in patients with metastatic nonsmall cell lung carcinoma: an Eastern Cooperative Oncology Group Study (E1587). Cancer 1998; 82: 292–300. source
- Interpretationown data
The results are consistent with a predictable pattern of inhibition of known HIF-1-inducible genes.
“they are consistent with a predictable pattern of inhibition of known HIF-1-inducible genes.”
Quote in the paperSubstantial claim - Interpretationreasoning
The sequence specificity of NSC-13502 may not be restricted to HIF-1 but may include other transcription factors that have similar core-binding sites in their DNA consensus sequences.
“It is, however, conceivable that the sequence specificity of NSC-13502 may not be restricted to HIF-1 but may include other transcription factors that have similar core-binding sites in their DNA consensus sequences.”
Quote in the paperNot a substantial claim - Premisecitation
Overexpression of HIF-1α has been implicated as a poor prognostic indicator in a variety of tumors.
“overexpression of HIF-1a has been implicated as a poor prognostic indicator in a variety of tumors (20)(21)(22)(23).”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [20] Zhong H, De Marzo AM, Laughner E, et al. Overexpression of hypoxia-inducible factor 1α in common human cancers and their metastases. Cancer Res 1999; 59: 5830–5.
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- cites [21] Shibaji T, Nagao M, Ikeda N, et al. Prognostic significance of HIF-1α overexpression in human pancreatic cancer. Anticancer Res 2003; 23: 4721–7.
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- cites [22] Bachtiary B, Schindl M, Potter R, et al. Overexpression of hypoxia-inducible factor 1α indicates diminished response to radiotherapy and unfavorable prognosis in patients receiving radical radiotherapy for cervical cancer. Clin Cancer Res 2003; 9: 2234–40.
- Premisecitation
NSC-13502, at 1.2 to 1.5 mg/m2 once weekly for 4 weeks or once every 3 to 4 weeks, has been extensively tested in phase I-II clinical trials.
“NSC-13502, at 1.2 to 1.5 mg/m 2 once weekly for 4 weeks or once every 3 to 4 weeks (47-50), has been extensively tested in phase I-II clinical trials.”
Quote in the paperNot a substantial claim0 of 4 sources checked- cites [47] Foster BJ, Clagett-Carr K, Shoemaker DD, et al. Echinomycin: the first bifunctional intercalating agent in clinical trials. Invest New Drugs 1985; 3: 403–10. source
- cites [48] Muss HB, Blessing JA, DuBeshter B. Echinomycin in recurrent and metastatic endometrial carcinoma. A phase II trial of the Gynecologic Oncology Group. Am J Clin Oncol 1993; 16: 492–3. source
- cites [49] Wadler S, Tenteromano L, Cazenave L, et al. Phase II trial of echinomycin in patients with advanced or recurrent colorectal cancer. Cancer Chemother Pharmacol 1994; 34: 266–9. source
- cites [50] Chang AY, Kim K, Boucher H, et al. A randomized phase II trial of echinomycin, trimetrexate, and cisplatin plus etoposide in patients with metastatic nonsmall cell lung carcinoma: an Eastern Cooperative Oncology Group Study (E1587). Cancer 1998; 82: 292–300. source
- Interpretationown data
NSC-13502 has a limited effect on global gene expression in U251 cells.
“Fewer genes were changed by z4-fold, suggesting that NSC-13502 has a limited effect on global gene expression (Fig. 5B).”
- Also stated in: § Discussion · plain
“Global analysis of gene expression using oligonucleotide arrays containing sequences representing f11,000 human genes showed that a relatively small number of genes are changed by addition of NSC-13502 under either normoxic or hypoxic conditions.”
- Also stated in: § Results · plain
“In conclusion, our results show that expression of a limited number of genes is affected by treatment of U251 cells with NSC-13502”
Quote in the paperSubstantial claim - Also stated in: § Discussion · plain
- Premisecitation
NSC-13502 is a small molecule known to bind DNA in a sequence-specific fashion.
“NSC-13502, a cyclic peptide of the family of quinoxaline antibiotics that was originally isolated from Streptomyces echinatus (45), is a small molecule known to bind DNA in a sequence-specific fashion.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [15] Van Dyke MM, Dervan PB. Echinomycin binding sites on DNA. Science 1984; 225: 1122–7. source
- Interpretationown data
NSC-13502 can inhibit, in a sequence-specific fashion, the binding of HIF-1 to its cognate DNA-binding sequence.
“We provide here proof-of-principle that NSC-13502 can in fact inhibit, in a sequence-specific fashion, the binding of HIF-1 to its cognate DNA-binding sequence.”
- Also stated in: § Discussion · boosted
“We believe that these results provide strong evidence that NSC-13502 interferes with HIF-1 DNA binding in a sequencespecific fashion.”
- Also stated in: § Discussion · plain
“NSC-13502 specifically inhibits HIF-1 DNA binding to the HRE.”
Quote in the paperSubstantial claim - Also stated in: § Discussion · boosted
- Premisecitation
Most activities of transcription factors are mediated by protein-protein interaction or binding to DNA in a sequence-specific fashion, both conventionally considered challenging targets for development of small-molecule inhibitors.
“However, most of the activities of transcription factors are mediated by protein-protein interaction or binding to DNA in a sequence-specific fashion, both of which are conventionally considered challenging targets for development of small-molecule inhibitors (2).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [2] Arkin MR, Wells JA. Small-molecule inhibitors of protein-protein interactions: progressing towards the dream. Nat Rev Drug Discov 2004; 3: 301–17. source
- Classified as probably not a substantial claim. 9
- May combine two claims. 17
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
PremisecitationA synthetic pyrrole-imidazole polyamide that inhibits in a sequence-specific fashion HIF binding to the HRE of the VEGF promoter caused 264 and 73 genes to be down-regulated and up-regulated by 2-fold, respectively, at a concentration of 1 μmol/L.
“Interestingly, a synthetic pyrroleimidazole polyamide that inhibits in a sequence-specific fashion HIF binding to the HRE of the VEGF promoter (44) caused 264 and 73 genes to be down-regulated and up-regulated by 2-fold, respectively, at a concentration of 1 Amol/L.”
Quote in the paperNot a substantial claim1 of 1 sources checked- cites [44] Olenyuk BZ, Zhang GJ, Klco JM, et al. Inhibition of vascular endothelial growth factor with a sequence-specific hypoxia response element antagonist. Proc Natl Acad Sci U S A 2004; 101: 16768–73. source
May combine two claims.
Findingown dataNSC-13502 causes approximately 50% displacement of ethidium bromide on HRE oligonucleotide at 3 μmol/L, whereas no significant influence on AP-1 oligonucleotide is observed even at 10 μmol/L.
“NSC-13502 caused f50% displacement of ethidium bromide on HRE oligonucleotide at 3 Amol/L, whereas no significant influence on AP-1 oligonucleotide was observed even at a concentration of 10 Amol/L (data not shown).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataNSC-13502 specifically inhibits HIF-1, but not AP-1 or NF-κB, binding activity to endogenous promoters in cultured cells.
“These results clearly show that NSC-13502 specifically inhibits HIF-1, but not AP-1 or NF-nB, binding activity to endogenous promoters in cultured cells.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown data65 genes are down-regulated by ≥4-fold under hypoxia and 36 under normoxia in U251 cells treated with NSC-13502.
“451 versus 276 by z2-fold and 65 versus 36 by z4-fold”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataNSC-13502 under the same experimental conditions did not block the binding of c-Fos to the AP-1 site of the cyclin D1 promoter or the binding of p65 to the NF-κB site of the ICAM-1 promoter.
“More importantly, NSC-13502 under the same experimental conditions did not block the binding of c-Fos to the AP-1 site of the cyclin D1 promoter or the binding of p65 to the NF-nB site of the ICAM-1 promoter.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataNSC-13502 up-regulates 186 genes under normoxia and 197 under hypoxia by ≥2-fold in U251 cells.
“only 186 genes were up-regulated under normoxia and 197 under hypoxia.”
- Also stated in: § Discussion · plain
“addition of NSC-13502 to U251 cells cultured under normoxic conditions only down-regulated or up-regulated 276 and 186 genes, respectively, by z2-fold.”
Quote in the paperNot a substantial claim- Also stated in: § Discussion · plain
May combine two claims.
Findingown dataNSC-13502 is one of the most potent compounds identified in the HIF-1 cell-based high-throughput screen and further validated using DNA-binding assays.
“Figure 2A shows the chemical structure of NSC-13502, one of the most potent compounds identified in the HIF-1 cell-based high-throughput screen and further validated using DNA-binding assays.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataHIF-1α mRNA expression is slightly decreased under hypoxic conditions relative to normoxia.
“In contrast, HIF-1a mRNA expression, which was slightly decreased under hypoxic conditions relative to normoxia, was not affected by NSC-13502 (data not shown).”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataNSC-13502 down-regulates 276 genes by ≥2-fold under normoxic conditions and 451 genes under hypoxic conditions in U251 cells.
“276 and 451 genes were down-regulated by z2-fold under normoxic or hypoxic conditions, respectively, in the presence of NSC-13502 (Fig. 5B).”
- Also stated in: § Discussion · plain
“addition of NSC-13502 to U251 cells cultured under normoxic conditions only down-regulated or up-regulated 276 and 186 genes, respectively, by z2-fold.”
Quote in the paperSubstantial claim- Also stated in: § Discussion · plain
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
PremisecitationHIF-1α and HIF-1β belong to the bHLH-PAS family; the basic domain is responsible for sequence-specific DNA binding, whereas HLH and PAS domains mediate dimerization and stabilization of the complex on DNA.
“HIF-1a and HIF-1h belong to the basic-helix-loop-helix (bHLH)-PAS family of transcription factors.The basic domain is responsible for sequence-specific DNA binding, whereas the HLH and the PAS domains mediate dimerization and stabilization of the complex on the DNA (12).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [12] Jiang BH, Rue E, Wang GL, Roe R, Semenza GL. Dimerization, DNA binding, and transactivation properties of hypoxia-inducible factor 1. J Biol Chem 1996; 271: 17771–8. source
Classified as probably not a substantial claim.
PremisecitationHIF-1 activates transcription of genes involved in cancer biology including cell survival, glycolysis, angiogenesis, migration, and invasion.
“HIF-1 activates the transcription of genes whose products are involved in crucial aspects of cancer biology, including cell survival, glycolysis, angiogenesis, migration, and invasion (5).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [5] Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer 2003; 3: 721–32. source
May combine two claims.
Interpretationown dataNSC-13502 inhibits HIF-1, but not AP-1 or NF-κB, DNA-binding activity in vitro by binding DNA in a sequence-specific fashion.
“Results shown thus far indicate that NSC-13502 inhibits HIF-1, but not AP-1 or NF-nB, DNA-binding activity in vitro by binding DNA in a sequence-specific fashion.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationThe sequence specificity of NSC-13502 encompasses the core sequence (5'-CGTG-3') of the HRE consensus sequence 5'-R(A/G)CGTG-3'.
“Interestingly, NSC-13502 was originally discovered as a sequence-specific DNA-binding agent (15), whose sequence specificity encompasses the core sequence (5V-CGTG-3V) of HRE consensus sequence 5V-R(A/ G)CGTG-3V (12).”
Quote in the paperNot a substantial claim0 of 2 sources checkedClassified as probably not a substantial claim. May combine two claims.
PremisecitationHIF-1 is a heterodimer composed of HIF-1α and HIF-1β (ARNT) that mediates the transcriptional program to oxygen deprivation.
“Hypoxia-inducible factor-1 (HIF-1), a heterodimer composed of two subunits, HIF-1a and HIF-1h (also known as aryl hydrocarbon receptor nuclear translocator), mediates the transcriptional program to oxygen deprivation (3).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [3] Semenza GL. Hypoxia-inducible factor 1: oxygen homeostasis and disease pathophysiology. Trends Mol Med 2001; 7: 345–50. source
May combine two claims.
PremisecitationTopotecan administered on a chronic schedule inhibits HIF-1α protein accumulation, angiogenesis, and tumor growth in U251-HRE human glioblastoma xenografts.
“More importantly, we have shown that topotecan administered on a chronic schedule inhibits HIF-1a protein accumulation, angiogenesis, and tumor growth in U251-HRE human glioblastoma xenografts (38).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [38] Rapisarda A, Zalek J, Hollingshead M, et al. Schedule-dependent inhibition of hypoxia-inducible factor-1α protein accumulation, angiogenesis, and tumor growth by topotecan in U251-HRE glioblastoma xenografts. Cancer Res 2004; 64: 6845–8. source
May combine two claims.
PremisecitationThe strong binding sites for NSC-13502 contain the central 2-bp sequence 5'-CG-3', and the key recognition elements for NSC-13502 are contained in the sequences 5'-ACGT-3' and 5'-TCGT-3'.
“The strong binding sites for NSC-13502 contain the central 2-bp sequence 5V-CG-3V, and the key recognition elements for NSC-13502 are contained in the sequences 5V-ACGT-3V and 5V-TCGT-3V (15).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [15] Van Dyke MM, Dervan PB. Echinomycin binding sites on DNA. Science 1984; 225: 1122–7. source
May combine two claims.
PremisecitationThe Myc/Max family DNA-binding site (E-box, 5'-CACGTG-3') encompasses the core sequence 5'-CGTG-3', which is also present in the HIF-1 consensus sequence and is part of the sequence specificity of NSC-13502.
“One example is the Myc/ Max family of transcription factors, whose DNA-binding site (E-box, 5V-CACGTG-3V; ref. 46) encompasses the core sequence 5V-CGTG-3V, which is also present in the HIF-1 consensus sequence and is part of the sequence specificity of NSC-13502.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [46] Nair SK, Burley SK. X-ray structures of Myc-Max and Mad-Max recognizing DNA. Molecular bases of regulation by proto-oncogenic transcription factors. Cell 2003; 112: 193–205. source
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationNSC-13502 was brought into clinical trials by the NCI 20 years ago based on its antitumor activity against two i.p. implanted murine tumors, the B16 melanoma and the P388 leukemia.
“NSC-13502 was brought into clinical trials by the NCI 20 years ago based on its antitumor activity against two i.p. implanted murine tumors, the B16 melanoma and the P388 leukemia (47).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [47] Foster BJ, Clagett-Carr K, Shoemaker DD, et al. Echinomycin: the first bifunctional intercalating agent in clinical trials. Invest New Drugs 1985; 3: 403–10. source
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationTopotecan inhibits HIF-1α protein accumulation independently of DNA replication-mediated DNA damage.
“Our group has recently shown that topotecan, a topoisomerase I poison, inhibits HIF-1a protein accumulation independently of DNA replication-mediated DNA damage, suggesting a distinct mechanism of action from the cytotoxic effects (35).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [35] Rapisarda A, Uranchimeg B, Sordet O, et al. Topoisomerase I-mediated inhibition of hypoxia-inducible factor 1: mechanism and therapeutic implications. Cancer Res 2004; 64: 1475–82. source
May combine two claims.
Interpretationown dataNSC-13502 binds DNA and not HIF-1α and/or HIF-1β proteins.
“consistent with the notion that NSC-13502 binds DNA and not HIF-1a and/or HIF-1h proteins”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
InterpretationreasoningMost agents described above affect targets that are also involved in other biological functions, which will ultimately determine their therapeutic activity, and they can be defined nonselective inhibitors of HIF-1.
“Most of the agents described above affect targets that are also involved in other biological functions, which will ultimately determine their therapeutic activity, and they can be defined nonselective inhibitors of HIF-1.”
Quote in the paperNot a substantial claimMay combine two claims.
PremisecitationHIF-1α is the downstream mediator of hypoxic and nonhypoxic signaling pathways, which play a crucial role in cancer progression.
“HIF-1a is the downstream mediator of hypoxic and nonhypoxic signaling pathways, which play a crucial role in cancer progression.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [5] Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer 2003; 3: 721–32. source
The authors state this as an assumption.
NSC-13502 inhibits HIF-1 DNA-binding activity in vitro by binding DNA in a sequence-specific fashion.
“Results shown thus far indicate that NSC-13502 inhibits HIF-1, but not AP-1 or NF-nB, DNA-binding activity in vitro by binding DNA in a sequence-specific fashion.If this assumption were correct, a ChIP assay would show whether NSC-13502 inhibited endogenous HIF-1 DNA-binding activity in cultured cells.”
The paper relies on this without support.
The core sequence of the HRE (5'-CGTG-3') is also present in the E-box (5'-CACGTG-3'), the DNA-binding site of the Myc/Max complex.
“The core sequence of the HRE (5V -CGTG-3V) is also present in the E-box (5V-CACGTG-3V), which is the DNA-binding site of the Myc/Max complex, raising the possibility that NSC-13502 might also inhibit the binding of nuclear proteins to the E-box.”
The paper relies on this without support.
The HRE recognized by HIF-1 contains the core sequence 5'-R(A/G)CGTG-3' and is present in promoters of HIF-1 target genes.
“The DNA-binding site recognized by HIF-1, known as hypoxia-responsive element (HRE), contains the core sequence 5V -R(A/G)CGTG-3V , which is present in promoters of HIF-1 target genes.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Echinomycin preferentially intercalates into DNA at sites containing the central 5′-CG-3′ dinucleotide, a critical recognition element within the HIF consensus HRE sequence (5′-ACGT-3′), thereby preventing binding of both HIF-1α and HIF-2α to their cognate response elements.
“Echinomycin preferentially intercalates into DNA at sites containing the central 5′-CG-3′ dinucleotide, a critical recognition element within the HIF consensus HRE sequence (5′-ACGT-3′), thereby preventing binding of both HIF-1α and HIF-2α to their cognate response elements [383,384].”
From HIF inhibition: Current strategies and clinical challenges (2026) · cites this paper as [383]
Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measuredEchinomycin exhibits limited specificity and a high propensity for off-target transcriptional effects.
“As a result, despite its potent ability to block HIF–DNA interactions, echinomycin exhibits limited specificity and a high propensity for off-target transcriptional effects.”
From HIF inhibition: Current strategies and clinical challenges (2026) · cites this paper as [383]
The Discussion (page not available) acknowledges that echinomycin may also inhibit transcription factors with HIF-like DNA-binding sites, which could detract from its target specificity. But the Results (page not available) describe a limited effect on global gene expression, not a high propensity for off-target transcriptional effects. (LLM, conf=0.88)
Experimental — accuracy not yet measuredEchinomycin also binds DNA motifs recognized by other transcription factors, such as AP-1, NF-κB, or members of the Myc/Max family, whose E-box binding site overlaps substantially with the HIF HRE core.
“Echinomycin also binds DNA motifs recognized by other transcription factors, such as AP-1, NF-κB, or members of the Myc/Max family, whose E-box binding site (5′-CACGTG-3′) overlaps substantially with the HIF HRE core [383,385].”
From HIF inhibition: Current strategies and clinical challenges (2026) · cites this paper as [383]
The Results and Discussion (page not available) identify the Myc/Max E-box sequence, show that it contains the HIF HRE core, and report that echinomycin abolished protein binding to an E-box oligonucleotide. The Results report small melting-temperature increases for AP-1 and NF-κB oligonucleotides, but emphasize echinomycin’s higher affinity for HRE DNA and find no inhibition of AP-1 or NF-κB protein binding. Thus, the Myc/Max portion is supported, while the unqualified suggestion that echinomycin also binds AP-1 and NF-κB motifs is stronger than this paper establishes. (LLM, conf=0.84)
Experimental — accuracy not yet measuredEchinomycin is a potent small-molecule inhibitor of HIF-1α that functions through sequence-specific binding to HRE to competitively inhibit HIF-1α binding to its target genes.
“Echinomycin is a potent small-molecule inhibitor of HIF-1α, which functions through sequence-specific binding to HRE to competitively inhibit HIF-1α binding to its target genes9.”
From Liposomal Formulation of HIF-1α Inhibitor Echinomycin Eliminates Established Metastases of Triple-negative Breast Cancer (2020) · cites this paper as [9]
The Results (page unavailable) identify echinomycin as a potent inhibitor of HIF-1 DNA binding and show that it binds HRE-containing DNA with relatively high affinity. They also show reduced binding of the HIF-1α/HIF-1β complex to HREs, but describe inhibition through interaction with DNA as a possibility rather than establishing a competitive mechanism. The claim overstates that mechanism and describes the target as HIF-1α alone. (LLM, conf=0.86)
Experimental — accuracy not yet measuredEchinomycin binds specifically to the core of the HIF-1 recognition sequence 5'-CGTG-3' precluding HIF-1 and DNA binding.
“It binds specifically to the core of the HIF-1 recognition sequence 5′–CGTG-3′ in a sequence-dependent fashion precluding HIF-1 and DNA binding.”
From HIF-1α pathway: role, regulation and intervention for cancer therapy (2015) · cites this paper as [49]
Echinomycin leads to partial restoration of insulin-induced tyrosine phosphorylation of insulin receptor in hypoxia.
“The use of echinomycin, an inhibitor that has been characterized to inhibit HIF activity without modulating HIF-α subunit protein levels (23), led also to partial restoration of insulin-induced tyrosine phosphorylation of the insulin receptor in hypoxia (data not shown).”
From Hypoxia decreases insulin signaling pathways in adipocytes. (2009) · cites this paper as [23]
Echinomycin can inhibit HIF complex-induced binding to HRE regions in promoters of various genes.
“the HIF complex-induced binding to HRE regions in promoters of various genes can be achieved by echinomycin [87].”
From Hypoxia Signaling in Cancer: From Basics to Clinical Practice (2021) · cites this paper as [87]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Protein identifier conflicts with the constructs described
MinorThe screening result names HIF-1h-389, but the expressed 389-residue construct is identified elsewhere as HIF-1a-389; HIF-1h is the 475-residue construct.
“Twenty of 128 compounds inhibited HIF-1h-389 and HIF-1h-475 DNA-binding activity by >50% relative to untreated control at the 10 Amol/L concentration (data not shown).”
“we expressed recombinant HIF-1a-bHLH-PAS-His (HIF-1a-389) and Flag-HIF-1h-bHLH-PAS (HIF-1h-475) proteins”
Cites
Targeting HIF-1 for cancer therapy
Semenza, G. L. (2003) · Nature Reviews Cancer · doi:10.1038/nrc1187 · Cited by 6,717 · see citing works
Wang, G. L., Jiang, B. H., Rue, E. A., & Semenza, G. L. (1995) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.92.12.5510 · Cited by 6,212 · see citing works
The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis
Maxwell, P. H., Wiesener, M. S., Chang, G.-W., Clifford, S. C., Vaux, E. C., Cockman, M. E., Wykoff, C. C., Pugh, C. W., Maher, E. R., & Ratcliffe, P. J. (1999) · Nature · doi:10.1038/20459 · Cited by 5,180 · see citing works
Monks, A. P., Scudiero, D. A., Skehan, P., Shoemaker, R. H., Paull, K. D., Vistica, D. T., Hose, C. D., Langley, J. W., Cronise, P., Vaigro-Wolff, A., et al. (1991) · JNCI Journal of the National Cancer Institute · doi:10.1093/jnci/83.11.757 · Cited by 3,305 · see citing works
Self-citation(1999) · PubMed · Cited by 2,204 · see citing works
Small-molecule inhibitors of protein–protein interactions: progressing towards the dream
Arkin, M. R., & Wells, J. A. (2004) · Nature Reviews Drug Discovery · doi:10.1038/nrd1343 · Cited by 1,705 · see citing works
(2000) · PubMed · Cited by 1,445 · see citing works
Laughner, E., Taghavi, P., Chiles, K., Mahon, P. C., & Semenza, G. L. (2001) · Molecular and Cellular Biology · doi:10.1128/mcb.21.12.3995-4004.2001 · Cited by 1,256 · cited 1× · see citing works
“Notably, HIF-1a is overexpressed in human cancers due to hypoxia-dependent and/or hypoxia-independent pathways (6, 18, 19), and overexpression of HIF-1a has been implicated as a poor prognostic indicator in a variety of tumors (20–23).”
BackgroundTranscription factors as targets for cancer therapy
Darnell, J. E. (2002) · Nature Reviews Cancer · doi:10.1038/nrc906 · Cited by 1,194 · see citing works
Hudson, C. C., Liu, M., Chiang, G. G., Otterness, D. M., Loomis, D. C., Kaper, F., Giaccia, A. J., & Abraham, R. T. (2002) · Molecular and Cellular Biology · doi:10.1128/mcb.22.20.7004-7014.2002 · Cited by 1,186 · see citing works
Dimerization, DNA Binding, and Transactivation Properties of Hypoxia-inducible Factor 1
Jiang, B.-H., Rue, E., Wang, G. L., Roe, R., & Semenza, G. L. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.30.17771 · Cited by 1,106 · see citing works
Regulation of tumor angiogenesis by p53-induced degradation of hypoxia-inducible factor 1alpha.
Ravi, R. K., Mookerjee, B., Bhujwalla, Z. M., Sutter, T. R., Artemov, D., Zeng, Q., Dillehay, L. E., Madan, A., Semenza, G. L., & Bedi, A. (2000) · PubMed Central · Cited by 1,099 · see citing works
Regulation of tumor angiogenesis by p53-induced degradation of hypoxia-inducible factor 1α
Ravi, R., Mookerjee, B., Bhujwalla, Z. M., Sutter, C. H., Artemov, D., Zeng, Q., Dillehay, L. E., Madan, A., Semenza, G. L., & Bedi, A. (2000) · Genes & Development · doi:10.1101/gad.14.1.34 · Cited by 967 · see citing works
Hypoxia-inducible factor 1: oxygen homeostasis and disease pathophysiology
Semenza, G. L. (2001) · Trends in Molecular Medicine · doi:10.1016/s1471-4914(01)02090-1 · Cited by 930 · see citing works
Signal transduction to hypoxia-inducible factor 1
Semenza, G. L. (2002) · Biochemical Pharmacology · doi:10.1016/s0006-2952(02)01168-1 · Cited by 860 · see citing works
Loss of <i>PTEN</i> facilitates HIF-1-mediated gene expression
Zundel, W., Schindler, C., Haas-Kogan, D., Koong, A., Kaper, F., Chen, E., Gottschalk, A. R., Ryan, H. E., Johnson, R. S., Jefferson, A. B., et al. (2000) · Genes & Development · doi:10.1101/gad.14.4.391 · Cited by 840 · see citing works
Fukuda, R., Hirota, K., Fan, F., Jung, Y. D., Ellis, L. M., & Semenza, G. L. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m203781200 · Cited by 815 · see citing works
Hsp90 inhibitors as novel cancer chemotherapeutic agents
Neckers, L. (2002) · Trends in Molecular Medicine · doi:10.1016/s1471-4914(02)02316-x · Cited by 677 · see citing works
HIF-1 as a target for drug development
Giaccia, A., Siim, B. G., & Johnson, R. S. (2003) · Nature Reviews Drug Discovery · doi:10.1038/nrd1199 · Cited by 631 · see citing works
X-Ray Structures of Myc-Max and Mad-Max Recognizing DNA
Nair, S. K., & Burley, S. K. (2003) · Cell · doi:10.1016/s0092-8674(02)01284-9 · Cited by 551 · see citing works
Suppression of tumor growth through disruption of hypoxia-inducible transcription
Kung, A. L., Wang, S., Klco, J. M., Kaelin, W. G., & Livingston, D. M. (2000) · Nature Medicine · doi:10.1038/82146 · Cited by 501 · see citing works
Bos, R., van der Groep, P., Greijer, A. E., Shvarts, A., Meijer, S., Pinedo, H. M., Semenza, G. L., van Diest, P. J., & van der Wall, E. (2003) · Cancer · doi:10.1002/cncr.11246 · Cited by 500 · see citing works
Small molecule blockade of transcriptional coactivation of the hypoxia-inducible factor pathway
Kung, A. L., Zabludoff, S. D., France, D. S., Freedman, S. J., Tanner, E. A., Vieira, A., Cornell-Kennon, S., Lee, J., Wang, B., Wang, J., et al. (2004) · Cancer Cell · doi:10.1016/j.ccr.2004.06.009 · Cited by 480 · see citing works
YC-1: A Potential Anticancer Drug Targeting Hypoxia-Inducible Factor 1
Yeo, E., Chun, Y., Cho, Y.-S., Kim, J., Lee, J.-C., Kim, M.-S., & Park, J.-W. (2003) · JNCI Journal of the National Cancer Institute · doi:10.1093/jnci/95.7.516 · Cited by 477 · see citing works
Structural basis for recruitment of CBP/p300 by hypoxia-inducible factor-1α
Freedman, S. J., Sun, Z., Poy, F., Kung, A. L., Livingston, D. M., Wagner, G., & Eck, M. J. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.082117899 · Cited by 457 · see citing works
(2002) · PubMed · Cited by 457 · see citing works
Self-citationTopoisomerase I-Mediated Inhibition of Hypoxia-Inducible Factor 1
Rapisarda, A., Uranchimeg, B., Sordet, O., Pommier, Y., Shoemaker, R. H., & Melillo, G. (2004) · Cancer Research · doi:10.1158/0008-5472.can-03-3139 · Cited by 301 · see citing works
Self-citationMabjeesh, N. J., Post, D. E., Willard, M. T., Kaur, B., Meir, E. G. V., Simons, J. W., & Zhong, H. (2002) · PubMed · Cited by 299 · see citing works
Welsh, S. J., Williams, R. R., Kirkpatrick, L., Paine-Murrieta, G. D., & Powis, G. (2004) · PubMed · Cited by 280 · see citing works
(2003) · PubMed · Cited by 250 · see citing works
HIF-1: A Target For Cancer, Ischemia and Inflammation—Too Good to be True?
Melillo, G. (2004) · Cell Cycle · doi:10.4161/cc.3.2.689 · Cited by 247 · see citing works
Self-citationWelsh, S., Williams, R., Kirkpatrick, L., Paine-Murrieta, G., & Powis, G. (2004) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.233.3.3 · Cited by 240 · see citing works
Identification of a Novel Small-Molecule Inhibitor of the Hypoxia-Inducible Factor 1 Pathway
Tan, C., de Noronha, R. G., Roecker, A. J., Pyrzyńska, B., Khwaja, F. W., Zhang, Z., Zhang, H., Teng, Q., Nicholson, A. C., Giannakakou, P., et al. (2005) · Cancer Research · doi:10.1158/0008-5472.605.65.2 · Cited by 225 · see citing works
Olenyuk, B. Z., Zhang, G.-J., Klco, J. M., Nickols, N. G., Kaelin, W. G., & Dervan, P. B. (2004) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0407617101 · Cited by 219 · see citing works
Echinomycin Binding Sites on DNA
Van Dyke, M. M., & Dervan, P. B. (1984) · Science · doi:10.1126/science.6089341 · Cited by 218 · see citing works
Rapisarda, A., Zalek, J., Hollingshead, M., Braunschweig, T., Uranchimeg, B., Bonomi, C. A., Borgel, S. D., Carter, J. P., Hewitt, S. M., Shoemaker, R. H., et al. (2004) · Cancer Research · doi:10.1158/0008-5472.can-04-2116 · Cited by 187 · see citing works
Self-citation(2003) · PubMed · Cited by 167 · see citing works
Patel, D. J. (1979) · Accounts of Chemical Research · doi:10.1021/ar50136a002 · Cited by 147 · see citing works
Sun, X., Kanwar, J. R., Leung, E. Y., Lehnert, K., Wang, D., & Krissansen, G. W. (2001) · Gene Therapy · doi:10.1038/sj.gt.3301388 · Cited by 146 · see citing works
Prognostic significance of HIF-1 alpha overexpression in human pancreatic cancer.
(2003) · PubMed · Cited by 138 · see citing works
Activated pp60c-Src Leads to Elevated Hypoxia-inducible Factor (HIF)-1α Expression under Normoxia
Karni, R., Dor, Y., Keshet, E., Meyuhas, O., & Levitzki, A. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m206141200 · Cited by 116 · see citing works
Zhang, Q., Zhang, Z., Rao, J. Y., Sato, J. D., Brown, J. J., Messadi, D. V., & Le, A. D. (2004) · International Journal of Cancer · doi:10.1002/ijc.20334 · Cited by 83 · see citing works
Allosteric Inhibition of Protein−DNA Complexes by Polyamide−Intercalator Conjugates
Fechter, E. J., & Dervan, P. B. (2003) · Journal of the American Chemical Society · doi:10.1021/ja030125e · Cited by 70 · see citing works
Echinomycin: The first bifunctional intercalating agent in clinical trials
Foster, B. J., Clagett-Carr, K., Shoemaker, D. D., Suffness, M., Plowman, J., Trissel, L. A., Grieshaber, C. K., & Leyland-Jones, B. (1985) · Investigational New Drugs · doi:10.1007/bf00170766 · Cited by 61 · cited 1× · see citing works
“implanted murine tumors, the B16 melanoma and the P388 leukemia (47).”
BackgroundKurebayashi, J., Otsuki, T., Kurosumi, M., Soga, S., Akinaga, S., & Sonoo, H. (2001) · Japanese Journal of Cancer Research · doi:10.1111/j.1349-7006.2001.tb02159.x · Cited by 60 · see citing works
PATEL, D. J. (1979) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1979.tb13265.x · Cited by 54 · see citing works
Phase II trial of echinomycin in patients with advanced or recurrent colorectal cancer
Wadler, S., Tenteromano, L., Cazenave, L., Sparano, J. A., Greenwald, E. S., Rozenblit, A., Kaleya, R., & Wiernik, P. H. (1994) · Cancer Chemotherapy and Pharmacology · doi:10.1007/bf00685088 · Cited by 43 · see citing works
Dibenzoylmethane, a natural dietary compound, induces HIF-1α and increases expression of VEGF
Mabjeesh, N. J., Willard, M. T., Harris, W. B., Sun, H.-Y., Wang, R., Zhong, H., Umbreit, J. N., & Simons, J. W. (2003) · Biochemical and Biophysical Research Communications · doi:10.1016/s0006-291x(03)00336-x · Cited by 37 · see citing works
Muss, H. B., Blessing, J. A., & DuBeshter, B. (1993) · American Journal of Clinical Oncology · doi:10.1097/00000421-199312000-00006 · Cited by 33 · see citing works
Effect of phosphate and amino acids on echinomycin biosynthesis by Streptomyces echinatus
Formica, J. V., & Waring, M. J. (1983) · Antimicrobial Agents and Chemotherapy · doi:10.1128/aac.24.5.735 · Cited by 24 · cited 1× · see citing works
“(45), is a small molecule known to bind DNA in a sequence-specific fashion.”
BackgroundPyridine Nucleotide‐Dependent Dehydrogenases
Scrutton, M. C. (1979) · FEBS Letters · doi:10.1016/0014-5793(79)80996-5 · Cited by 21 · see citing works
Inhibition of major groove DNA binding bZIP proteins by positive patch polyamides.
Bremer, R., Wurtz, N. R., Szewczyk, J., & Dervan, P. (2001) · Bioorganic & Medicinal Chemistry · doi:10.1016/s0968-0896(01)00122-5 · Cited by 19 · see citing works
Chang, A. Y., Kim, K., Boucher, H., Bonomi, P., Stewart, J. A., Karp, D. D., & Blum, R. H. (1998) · Cancer · doi:10.1002/(sici)1097-0142(19980115)82:2
Phase II trial of echinomycin in patients with advanced or recurrent colorectal cancer
Wadler, S., Tenteromano, L., Cazenave, L., et al. (1994) · Cancer Chemotherapy and Pharmacology · doi:10.1007/s002800050139
Rapisarda, A., Uranchimeg, B., Hollingshead, M., Zalek, J., Bonomi, C. A., Borgel, S. D., Carter, J., Shoemaker, R., & Melillo, G. (2004) · European Journal of Cancer Supplements · doi:10.1016/s1359-6349(04)80334-3
Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Tumor cell metabolism: cancer's Achilles' heel
Kroemer, G., & Pouyssegur, J. (2008) · Cancer Cell · doi:10.1016/j.ccr.2008.05.005 · Cited by 2,337 · see citing works
HIF-1α pathway: role, regulation and intervention for cancer therapy
Masoud, G. N., & Li, W. (2015) · Acta Pharmaceutica Sinica B · doi:10.1016/j.apsb.2015.05.007 · Cited by 1,871 · see citing works
Defining the role of hypoxia-inducible factor 1 in cancer biology and therapeutics
Semenza, G. L. (2010) · Oncogene · doi:10.1038/onc.2009.441 · Cited by 1,712 · see citing works
“intercalating drugs, including echinomycin (Kong et al., 2005) and anthracyclines, such as doxorubicin (Adriamycin) and daunorubicin (Lee et al.”
Hypoxia signalling in cancer and approaches to enforce tumour regression
Pouysségur, J., Dayan, F., & Mazure, N. M. (2006) · Nature · doi:10.1038/nature04871 · Cited by 1,676 · see citing works
Hypoxia-Induced Angiogenesis: Good and Evil
Krock, B. L., Skuli, N., & Simon, M. C. (2011) · Genes & Cancer · doi:10.1177/1947601911423654 · Cited by 1,179 · see citing works
Energy metabolism in tumor cells
Moreno‐Sánchez, R., Rodríguez‐Enríquez, S., Marín‐Hernández, Á., & Saavedra, E. (2007) · FEBS Journal · doi:10.1111/j.1742-4658.2007.05686.x · Cited by 1,086 · see citing works
Therapeutic targeting of hypoxia and hypoxia-inducible factors in cancer
Wigerup, C., Påhlman, S., & Bexell, D. (2016) · Pharmacology & Therapeutics · doi:10.1016/j.pharmthera.2016.04.009 · Cited by 687 · see citing works
“Echinomycin is a small antibiotic molecule isolated from Streptomyces echinatus that binds DNA in a sequence-specific manner.”
BackgroundInfluentialTriterpenoids and rexinoids as multifunctional agents for the prevention and treatment of cancer
Liby, K. T., Yore, M. M., & Sporn, M. B. (2007) · Nature Reviews Cancer · doi:10.1038/nrc2129 · Cited by 637 · see citing works
Rohwer, N., & Cramer, T. (2011) · Drug Resistance Updates · doi:10.1016/j.drup.2011.03.001 · Cited by 608 · see citing works
Hypoxia-inducible factors: cancer progression and clinical translation
Wicks, E. E., & Semenza, G. L. (2022) · Journal of Clinical Investigation · doi:10.1172/jci159839 · Cited by 600 · see citing works
“HIF inhibitors that have shown antitumor activity in mouse tumor models include acriflavine (47), digoxin (106), echinomycin (156), the HSP90 inhibitors 17-allylaminogeldanamycin (157) and ganetespib (158), 2-methoxyestradiol (159), PX-478 (160), and YC-1 (161).”
BackgroundCancer Cell Metabolism in Hypoxia: Role of HIF-1 as Key Regulator and Therapeutic Target
Infantino, V., Santarsiero, A., Convertini, P., Todisco, S., & Iacobazzi, V. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms22115703 · Cited by 510 · see citing works
“doxorubicin, impair binding of HIF to chromatin, resulting in blocking of its transcriptional activity [146,147].”
BackgroundN‐Methylation of Peptides and Proteins: An Important Element for Modulating Biological Functions
Chatterjee, J., Rechenmacher, F., & Kessler, H. (2012) · Angewandte Chemie International Edition · doi:10.1002/anie.201205674 · Cited by 500 · see citing works
Angiogenesis Inhibitors: Current Strategies and Future Prospects
Cook, K. M., & Figg, W. D. (2010) · CA A Cancer Journal for Clinicians · doi:10.3322/caac.20075 · Cited by 478 · see citing works
Hypoxia-inducible factors: central regulators of the tumor phenotype
Gordan, J. D., & Simon, M. C. (2007) · Current Opinion in Genetics & Development · doi:10.1016/j.gde.2006.12.006 · Cited by 450 · see citing works
Autophagy in cancer associated fibroblasts promotes tumor cell survival
Martinez‐Outschoorn, U., Trimmer, C., Zhao, L., Whitaker‐Menezes, D., Chiavarina, B., Zhou, J., Wang, C., Pavlides, S., Cantarin, M. P. M., Capozza, F., et al. (2010) · Cell Cycle · doi:10.4161/cc.9.17.12928 · Cited by 406 · see citing works
Targeting HIF1α Eliminates Cancer Stem Cells in Hematological Malignancies
Wang, Y., Liu, Y., Malek, S. N., Zheng, P., & Liu, Y. (2011) · Cell stem cell · doi:10.1016/j.stem.2011.02.006 · Cited by 410 · see citing works
Endoplasmic Reticulum Stress and Cancer
Yadav, R. K., Chae, S., Kim, H., & Chae, H. (2014) · Journal of Cancer Prevention · doi:10.15430/jcp.2014.19.2.75 · Cited by 396 · see citing works
“Therefore, blocking the HIF1α signal might be a novel and promising therapeutic target for the treatment of hypoxic tumors.(21)”
BackgroundGLUT1 Expression Is Increased in Hepatocellular Carcinoma and Promotes Tumorigenesis
Amann, T., Maegdefrau, U., Hartmann, A., Agaimy, A., Marienhagen, J., Weiß, T. S., Stoeltzing, O., Warnecke, C., Schölmerich, J., Oefner, P. J., et al. (2009) · American Journal Of Pathology · doi:10.2353/ajpath.2009.080596 · Cited by 370 · see citing works
Targeting Transcription Factors for Cancer Treatment
Lambert, M., Jambon, S., Depauw, S., & David‐Cordonnier, M. (2018) · Molecules · doi:10.3390/molecules23061479 · Cited by 351 · see citing works
“In terms of transcription factor inhibition, echinomycin was identified as a HIF1α and HIF1β/DNA binding inhibitor as evidenced using EMSA, luciferase activity and ChIP experiments on HIF-1 response elements (HRE) of the vascular endothelial growth factor promoter (VEGF, associated with tumor vascularization) [277].”
MethodTargeting hypoxia signalling for the treatment of ischaemic and inflammatory diseases
Eltzschig, H. K., Bratton, D. L., & Colgan, S. P. (2014) · Nature Reviews Drug Discovery · doi:10.1038/nrd4422 · Cited by 363 · see citing works
Cairns, R. A., Papandreou, I., & Denko, N. C. (2006) · Molecular Cancer Research · doi:10.1158/1541-7786.mcr-06-0002 · Cited by 370 · see citing works
“Several small-molecule inhibitors of HIF have been identified (119-122) and are now being tested for antitumor activity in preclinical trials.”
BackgroundEvaluation of HIF-1 inhibitors as anticancer agents
Semenza, G. L. (2007) · Drug Discovery Today · doi:10.1016/j.drudis.2007.08.006 · Cited by 363 · see citing works
Mitophagy defects arising from BNip3 loss promote mammary tumor progression to metastasis
Chourasia, A. H., Tracy, K., Frankenberger, C. A., Boland, M. L., Sharifi, M. N., Drake, L. E., Sachleben, J. R., Asara, J. M., Locasale, J. W., Karczmar, G. S., et al. (2015) · EMBO Reports · doi:10.15252/embr.201540759 · Cited by 332 · see citing works
“To assess to what extent elevated Hif levels contributed to the observed hyper-proliferative phenotype (Fig 1H), we treated cells with 10 ng/ml echinomycin (a small molecule inhibitor of Hif DNA binding activity) [25] and confirmed that echinomycin reduced expression of key Hif target genes (Fig 5E).”
Up-down regulation of HIF-1α in cancer progression
Rashid, M., Zadeh, L. R., Baradaran, B., Molavi, O., Ghesmati, Z., Sabzichi, M., & Ramezani, F. (2021) · Gene · doi:10.1016/j.gene.2021.145796 · Cited by 291 · see citing works
Jensen, R. L. (2009) · Journal of Neuro-Oncology · doi:10.1007/s11060-009-9827-2 · Cited by 286 · see citing works
“Using similar methods, we have been working on developing HRE-controlled expression of short hairpin vector-expressing inhibitory RNA for specific posttran- scriptional gene silencing (Fig.”
BackgroundInfluentialPharmacologic Targeting of Hypoxia-Inducible Factors
Semenza, G. L. (2019) · Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev-pharmtox-010818-021637 · Cited by 288 · see citing works
Hypoxia decreases insulin signaling pathways in adipocytes.
Regazzetti, C., Peraldi, P., Grémeaux, T., Najem-Lendom, R., Ben-Sahra, I., Cormont, M., Bost, F., Le Marchand-Brustel, Y., Tanti, J.-F., & Giorgetti-Peraldi, S. (2009) · Diabetes · doi:10.2337/db08-0457 · Cited by 281 · see citing works
“The use of echinomycin, an inhibitor that has been characterized to inhibit HIF activity without modulating HIF- subunit protein levels (23), led also to partial nm ol D O G /m g pr ot /3 m in”
Lactate Activates HIF-1 in Oxidative but Not in Warburg-Phenotype Human Tumor Cells
Saedeleer, C. J. D., Copetti, T., Porporato, P. E., Verrax, J., Féron, O., & Sonveaux, P. (2012) · PLoS ONE · doi:10.1371/journal.pone.0046571 · Cited by 274 · see citing works
Recent advances in hypoxia-inducible factor (HIF)-1 inhibitors
Xia, Y., Choi, H.-K., & Lee, K. (2012) · European Journal of Medicinal Chemistry · doi:10.1016/j.ejmech.2012.01.033 · Cited by 278 · see citing works
Targeting Protein–Protein Interactions for Therapeutic Intervention: A Challenge for the Future
Zinzalla, G., & Thurston, D. E. (2009) · Future Medicinal Chemistry · doi:10.4155/fmc.09.12 · Cited by 257 · see citing works
Qiu, G., Jin, M., Dai, J., Sun, W., Feng, J., & Jin, W. (2017) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2017.05.002 · Cited by 245 · see citing works
Development of HIF‐1 inhibitors for cancer therapy
Onnis, B., Rapisarda, A., & Melillo, G. (2009) · Journal of Cellular and Molecular Medicine · doi:10.1111/j.1582-4934.2009.00876.x · Cited by 268 · see citing works
Self-citationPathania, D., Millard, M., & Neamati, N. (2009) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2009.05.010 · Cited by 265 · see citing works
Broekgaarden, M., Weijer, R., van Gulik, T. M., Hamblin, M. R., & Heger, M. (2015) · Cancer and Metastasis Reviews · doi:10.1007/s10555-015-9588-7 · Cited by 261 · see citing works
Inhibition of mitophagy drives macrophage activation and antibacterial defense during sepsis
Patoli, D., Mignotte, F., Deckert, V., Dusuel, A., Dumont, A., Rieu, A., Jalil, A., Dongen, K. V., Bourgeois, T., Gautier, T., et al. (2020) · Journal of Clinical Investigation · doi:10.1172/jci130996 · Cited by 248 · see citing works
“Cheng SC, et al. mTOR- and HIF-1α-mediated aerobic glycolysis as metabolic basis for trained immunity.”
BackgroundInfluentialTargeting hypoxia cell signaling for cancer therapy
Melillo, G. (2007) · Cancer and Metastasis Reviews · doi:10.1007/s10555-007-9059-x · Cited by 230 · see citing works
“Echinomycin was found to potently inhibit HIF-1 DNA binding activities both in ELISA and EMSA [ 91 ].”
BackgroundSelf-citationMolecular mechanisms mediating metastasis of hypoxic breast cancer cells
Semenza, G. L. (2012) · Trends in Molecular Medicine · doi:10.1016/j.molmed.2012.08.001 · Cited by 217 · see citing works
Metabolic control of the Treg/Th17 axis
Barbi, J. J., Pardoll, D. M., & Pan, F. (2013) · Immunological Reviews · doi:10.1111/imr.12029 · Cited by 215 · see citing works
“For instance, the well-characterized HIF-1 inhibiting compounds digoxin, acriflavine, and echinomycin suppress HIF-1 activity by reducing its expression (55), preventing HIF-1a/HIF-1b dimer formation (56), and interfering with HIF-1 DNA binding (58, 59), respectively.”
BackgroundTargeting the hypoxia-inducible factor (HIF) pathway in cancer
Poon, E., Harris, A. L., & Ashcroft, M. A. (2009) · Expert Reviews in Molecular Medicine · doi:10.1017/s1462399409001173 · Cited by 210 · see citing works
The hypoxic microenvironment: A determinant of cancer stem cell evolution
Carnero, A., & LLeonart, M. E. (2016) · BioEssays · doi:10.1002/bies.201670911 · Cited by 201 · see citing works
Kang, J., Kim, E., Kim, W., Seong, K. M., Youn, H., Kim, J. W., Kim, J., & Youn, B. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m113.490482 · Cited by 188 · see citing works
Development of Inhibitors Targeting Hypoxia-Inducible Factor 1 and 2 for Cancer Therapy
Yu, T., Tang, B., & Sun, X. (2017) · Yonsei Medical Journal · doi:10.3349/ymj.2017.58.3.489 · Cited by 192 · see citing works
“Echinomycin (quinomycin A), a small molecule known to bind DNA in a sequence-specific fashion, was shown to inhibit the binding of HIF-1 to the core HRE sequences (5’-CGTG-3’) contained in the VEGF promoter.(54,55)”
HIFs, angiogenesis, and cancer
Yang, Y., Sun, M., Wang, L., & Jiao, B. (2012) · Journal of Cellular Biochemistry · doi:10.1002/jcb.24438 · Cited by 189 · see citing works
A complex task? Direct modulation of transcription factors with small molecules
Koehler, A. N. (2010) · Current Opinion in Chemical Biology · doi:10.1016/j.cbpa.2010.03.022 · Cited by 189 · see citing works
“The compounds include the natural product echinomycin [66], a potent yet nonspecific intercalator that blocks HIF-1 DNA binding and transcription, and NSC 50253 [67], a modest inhibitor of PAS-A domain interactions involving HIF-1α and HIF-1β.”
BackgroundInterfering with Tumor Hypoxia for Radiotherapy Optimization
Telarović, I., Wenger, R. H., & Pruschy, M. N. (2021) · Journal of Experimental & Clinical Cancer Research · doi:10.1186/s13046-021-02000-x · Cited by 196 · see citing works
Metabolic targeting of hypoxia and HIF1 in solid tumors can enhance cytotoxic chemotherapy
Cairns, R. A., Papandreou, I., Sutphin, P. D., & Denko, N. C. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0611662104 · Cited by 182 · see citing works
Yang, L., Lin, C., Wang, L., Guo, H., & Wang, X. (2012) · Experimental Cell Research · doi:10.1016/j.yexcr.2012.07.017 · Cited by 179 · see citing works
HIF1A Reduces Acute Lung Injury by Optimizing Carbohydrate Metabolism in the Alveolar Epithelium
Eckle, T. D. L. G., Brodsky, K. S., Bonney, M., Packard, T. A., Han, J., Borchers, C. H., Mariani, T. J., Kominsky, D. J., Mittelbronn, M. G. A., & Eltzschig, H. K. (2013) · PLoS Biology · doi:10.1371/journal.pbio.1001665 · Cited by 165 · see citing works
Inhibiting Hypoxia-Inducible Factor 1 for Cancer Therapy
Melillo, G. (2006) · Molecular Cancer Research · doi:10.1158/1541-7786.mcr-06-0235 · Cited by 165 · see citing works
“Perhaps not unexpectedly, only a few examples of HIF inhibitors that potentially target selective pathways associated with HIF activation have been described, among which echinomycin (29) and synthetic polyamides (30) that inhibit HIF-1 DNA binding, and chetomin (31), which blocks recruitment of coactivator p300/CBP (…”
BackgroundSelf-citationHypoxia-inducible factor pathway and diseases of the vascular wall
Lim, C. S., Kiriakidis, S., Sandison, A., Paleolog, E. M., & Davies, A. H. (2013) · Journal of Vascular Surgery · doi:10.1016/j.jvs.2013.02.240 · Cited by 154 · see citing works
“Examples of HIF inhibitors include EZN-2968,(132) sorafenib,(131) and echinomycin.(133) On the other hand, drugs that increase the activity of HIF-a are being developed for diseases such as ischemic heart diseases, stroke, peripheral arterial diseases, and chronic anemia.”
BackgroundMahabeleshwar, G. H., Kawanami, D., Sharma, N., Takami, Y., Zhou, G., Shi, H., Nayak, L. V., Jeyaraj, D., Grealy, R., White, M., et al. (2011) · Immunity · doi:10.1016/j.immuni.2011.04.014 · Cited by 149 · see citing works
“Echinomycin is a well-established inhibitor of HIF-1α (Kong et al., 2005).”
Maybin, J. A., Murray, A. A., Saunders, P. T. K., Hirani, N., Carmeliet, P., & Critchley, H. (2018) · Nature Communications · doi:10.1038/s41467-017-02375-6 · Cited by 149 · see citing works
Vadlapatla, R. K., Vadlapudi, A. D., & Mitra, A. K. (2013) · Current Drug Targets · doi:10.2174/13894501113149990015 · Cited by 147 · see citing works
ROS-activated Anticancer Prodrugs: a New Strategy for tumor-specific Damage
Peng, X., & Gandhi, V. V. (2012) · Therapeutic Delivery · doi:10.4155/tde.12.61 · Cited by 147 · see citing works
Hypoxia-induced alternative splicing: the 11th Hallmark of Cancer
Farina, A. R., Cappabianca, L. A., Sebastiano, M., Zelli, V., Guadagni, S., & Mackay, A. R. (2020) · Journal of Experimental & Clinical Cancer Research · doi:10.1186/s13046-020-01616-9 · Cited by 131 · see citing works
Hypoxia Contributes to Melanoma Heterogeneity by Triggering HIF1α-Dependent Phenotype Switching
Widmer, D. S., Hoek, K. S., Cheng, P. F., Eichhoff, O. M., Biedermann, T., Raaijmakers, M. I. G., Hemmi, S., Dummer, R., & Levesque, M. P. (2013) · Journal of Investigative Dermatology · doi:10.1038/jid.2013.115 · Cited by 139 · see citing works
“To inhibit HIF1a transcriptional activity, we treated the cells with Echinomycin during exposure to hypoxia (1% O2 for 72 hours) (Kong et al., 2005).”
MethodHypoxia-Inducible Factor-1: A Critical Player in the Survival Strategy of Stressed Cells
Chen, S., & Sang, N. (2015) · Journal of Cellular Biochemistry · doi:10.1002/jcb.25283 · Cited by 135 · see citing works
“, 2005]; PX478 and 103D5R showed the ability to inhibit HIF-1 function through unkown mechanisms [Welsh et al., 2004; Kong et al., 2005; Tan et al., 2005].”
BackgroundInfluentialPacary, E., Legros, H., Valable, S., Duchatelle, P., Lecocq, M., Petit, E., Nicole, O., & Bernaudin, M. (2006) · Journal of Cell Science · doi:10.1242/jcs.03004 · Cited by 143 · see citing works
“In addition, we used echinomycin, a small molecule previously reported by Kong et al. (Kong et al., 2005) which inhibits HIF- 1 DNA-binding activity.”
BackgroundMethodRole of the hypoxic tumor microenvironment in the resistance to anti-angiogenic therapies
Rapisarda, A., & Melillo, G. (2009) · Drug Resistance Updates · doi:10.1016/j.drup.2009.03.002 · Cited by 141 · see citing works
“, 2008) or the AhR ligand aminoflavone (Terzuoli and Melillo, submitted for publication) and inhibition of HIF-1 DNA binding by echinomycin (Kong et al., 2005) and doxorubicin (Lee et al.”
BackgroundSelf-citationHypoxia Inducible Factor Pathway Inhibitors as Anticancer Therapeutics
Burroughs, S. K., Kaluz, S., Wang, D., Wang, K., Van Meir, E. G., & Wang, B. (2013) · Future Medicinal Chemistry · doi:10.4155/fmc.13.17 · Cited by 140 · see citing works
Role and potential for therapeutic targeting of MYB in leukemia
Pattabiraman, D. R., & Gonda, T. J. (2012) · Leukemia · doi:10.1038/leu.2012.225 · Cited by 134 · see citing works
Hypoxia and Mucosal Inflammation
Colgan, S. P., Campbell, E. L., & Kominsky, D. J. (2016) · Annual Review of Pathology Mechanisms of Disease · doi:10.1146/annurev-pathol-012615-044231 · Cited by 133 · see citing works
Signaling by Hypoxia-Inducible Factors Is Critical for Ovulation In Mice
Kim, J., Bagchi, I. C., & Bagchi, M. K. (2009) · Endocrinology · doi:10.1210/en.2008-0948 · Cited by 134 · see citing works
“Echinomycin is a sequence-specific DNA-binding agent that binds the sequence 5 -CGTG-3 , which forms the core of the HRE [5 -(A/G)CGTG-3 ], which is found at the promoters of HIF-target genes (22).”
BackgroundInfluentialHai, Y., Wei, M., Wang, C., Gu, Y., & Shao, C. (2021) · Marine Life Science & Technology · doi:10.1007/s42995-021-00101-2 · Cited by 133 · see citing works
Wu, D., Zhau, H. E., Huang, W.-C., Iqbal, S. A., Habib, F. K., Sartor, O. A., Cvitanovic, L., Marshall, F. F., Xu, Z., & Chung, L. W. K. (2007) · Oncogene · doi:10.1038/sj.onc.1210316 · Cited by 132 · see citing works
“From the isolated DNA, a 135-bp region was amplified by PCR (Kong et al., 2005).”
BackgroundMethodS-Adenosylmethionine: more than just a methyl donor
Lee, Y., Ren, D., Jeon, B., & Liu, H. (2023) · Natural Product Reports · doi:10.1039/d2np00086e · Cited by 128 · see citing works
Marine antifungal theonellamides target 3β-hydroxysterol to activate Rho1 signaling
Nishimura, S., Arita, Y., Honda, M., Iwamoto, K., Matsuyama, A., Shirai, A., Kawasaki, H., Kakeya, H., Kobayashi, T., Matsunaga, S., et al. (2010) · Nature Chemical Biology · doi:10.1038/nchembio.387 · Cited by 125 · see citing works
Mitochondria as targets for chemotherapy
Gogvadze, V. G., Orrenius, S., & Zhivotovsky, B. (2009) · APOPTOSIS · doi:10.1007/s10495-009-0323-0 · Cited by 125 · see citing works
“Echinomycin HIF1 Inhibition achieved by DNA binding [ 119 ]”
MethodModulating Hypoxia-Inducible Transcription by Disrupting the HIF-1–DNA Interface
Nickols, N. G., Jacobs, C. S., Farkas, M. E., & Dervan, P. B. (2007) · ACS Chemical Biology · doi:10.1021/cb700110z · Cited by 127 · see citing works
“As previously reported, 100 nM echinomycin potently inhibits VEGF expression to levels below the non-induced control (21).”
BackgroundInfluentialHypoxia-inducible factors in cancer stem cells and inflammation
Peng, G., & Liu, Y. (2015) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2015.03.003 · Cited by 124 · see citing works
“This HRE sequence was specifically targeted by echinomycin [9, 42].”
The potential role of HIF on tumour progression and dissemination
Unwith, S., Zhao, H., Hennah, L., & Ma, D. (2014) · International Journal of Cancer · doi:10.1002/ijc.28889 · Cited by 126 · see citing works
Zou, G.-M., & Maitra, A. (2008) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-08-0113 · Cited by 124 · see citing works
Zhang, J., Yao, M., Xia, S., Zeng, F., & Liu, Q. (2025) · Cellular & Molecular Biology Letters · doi:10.1186/s11658-024-00682-7 · Cited by 139 · see citing works
Hsu, S.-H., Chen, C.-T., & Wei, Y. (2013) · Stem Cells · doi:10.1002/stem.1441 · Cited by 122 · see citing works
“To further confirm the involvement of HIF-1 , we treated hMSCs with echinomycin, a small-molecule inhibitor of HIF-1 transcription activity [19], during differentiation under hypoxia.”
MethodTsai, J. M., Sinha, R., Seita, J., Fernhoff, N. B., Christ, S., Koopmans, T., Krampitz, G. W., McKenna, K. M., Xing, L., Sandholzer, M., et al. (2018) · Science Translational Medicine · doi:10.1126/scitranslmed.aan6735 · Cited by 120 · see citing works
Real-Time Imaging of HIF-1α Stabilization and Degradation
Moroz, E., Carlin, S., Dyomina, K., Burke, S. A., Thaler, H. T., Blasberg, R., & Serganova, I. S. (2009) · PLoS ONE · doi:10.1371/journal.pone.0005077 · Cited by 117 · see citing works
The magic of the hypoxia-signaling cascade
Benizri, E. I., Ginouvès, A., & Berra, E. (2008) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-008-7472-0 · Cited by 111 · see citing works
Development of novel therapeutic strategies that target HIF-1
Semenza, G. L. (2006) · Expert Opinion on Therapeutic Targets · doi:10.1517/14728222.10.2.267 · Cited by 120 · see citing works
“In cancer cells, the transcription of many tumour suppressor genes is turned off through epigenetic regulation of histone acetylation and DNA methylation [115].”
BackgroundInfluentialGlioblastoma, Cancer Stem Cells and Hypoxia
Bar, E. E. (2010) · Brain Pathology · doi:10.1111/j.1750-3639.2010.00460.x · Cited by 117 · see citing works
Hypoxia, drug therapy and toxicity
Lee, K., Roth, R. A., & LaPres, J. J. (2006) · Pharmacology & Therapeutics · doi:10.1016/j.pharmthera.2006.08.001 · Cited by 118 · see citing works
Targeting Intrinsically Disordered Transcription Factors: Changing the Paradigm
Tsafou, K., Tiwari, P. B., Forman-Kay, J. D., Metallo, S. J., & Toretsky, J. A. (2018) · Journal of Molecular Biology · doi:10.1016/j.jmb.2018.04.008 · Cited by 110 · see citing works
Hypoxia signaling in cancer: Implications for therapeutic interventions
Zhuang, Y., Liu, K., He, Q., Gu, X., Jiang, C., & Wu, J. (2023) · MedComm · doi:10.1002/mco2.203 · Cited by 119 · see citing works
Hypoxia inducible factor-1: a novel target for cancer therapy
Belozerov, V. E., & Meir, E. G. V. (2005) · Anti-Cancer Drugs · doi:10.1097/01.cad.0000180116.85912.69 · Cited by 117 · see citing works
Inhibition of transcription factors with small organic molecules
Berg, T. (2008) · Current Opinion in Chemical Biology · doi:10.1016/j.cbpa.2008.07.023 · Cited by 115 · see citing works
Baumeister, J., Chatain, N., Hubrich, A., Maié, T., Costa, I. G., Denecke, B., Han, L., Küstermann, C., Sontag, S., Seré, K. M., et al. (2019) · Leukemia · doi:10.1038/s41375-019-0629-z · Cited by 86 · see citing works
“While HIF-2α protein was detectable and gradually increasing between 48 and 96 h of cultivation up to a level of shHIF-1α transduced cells, no increase by echinomycin was found (Supplementary Fig.”
BackgroundInfluentialA chemical genomics screen highlights the essential role of mitochondria in HIF-1 regulation
Lin, X., David, C. A., Donnelly, J. B., Michaelides, M. R., Chandel, N. S., Huang, X., Warrior, U., Weinberg, F. D., Tormos, K. V., Fesik, S. W., et al. (2008) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0706585104 · Cited by 115 · see citing works
Targeting cancer cells through autophagy for anticancer therapy
Turcotte, S., & Giaccia, A. J. (2010) · Current Opinion in Cell Biology · doi:10.1016/j.ceb.2009.12.007 · Cited by 114 · see citing works
Recent agents targeting HIF‐1α for cancer therapy
Hu, Y., Liu, J., & Huang, H. (2013) · Journal of Cellular Biochemistry · doi:10.1002/jcb.24390 · Cited by 113 · see citing works
“specifically binds to the core of the HIF-1a recognition sequence and inhibits binding of HIF-1 to the HRE sequence [Kong, 2005].”
BackgroundDuette, G., Gerber, P. P., Rubione, J., Pérez, P. S., LANDAY, A. L., Crowe, S. M., Liao, Z., Witwer, K. W., Holgado, M. P., Salido, J., et al. (2018) · mBio · doi:10.1128/mbio.00757-18 · Cited by 107 · see citing works
Terzuoli, E., Puppo, M., Rapisarda, A., Uranchimeg, B., Cao, L., Burger, A. M., Ziche, M., & Melillo, G. (2010) · Cancer Research · doi:10.1158/0008-5472.can-10-1075 · Cited by 111 · see citing works
“We and others have identified small molecule inhibitors of HIF-1α that act by distinct mechanisms of action, including, but not limited to, inhibition of HIF-1α translation (31,34,37,38), inhibition of HIF-1 DNA binding (39) or transcriptional activity (40–42), inhibition of protein-protein interaction (43), increased…”
Self-citationHypoxia Signaling in Cancer: From Basics to Clinical Practice
Sebestyén, A., Kopper, L., Dankó, T., & Tímár, J. (2021) · Pathology and Oncology Research · doi:10.3389/pore.2021.1609802 · Cited by 110 · see citing works
“Of note, the HIF complex-induced binding to HRE regions in promoters of various genes can be achieved by echinomycin [87].”
BackgroundTargeting HIF-2 α in clear cell renal cell carcinoma: A promising therapeutic strategy
Martínez‐Sáez, O., Borau, P. G., Alonso‐Gordoa, T., Molina‐Cerrillo, J., & Grande, E. (2017) · Critical Reviews in Oncology/Hematology · doi:10.1016/j.critrevonc.2017.01.013 · Cited by 109 · see citing works
Belaïdi, E., Morand, J., Gras, E., Pépin, J., & Godin‐Ribuot, D. (2016) · Pharmacology & Therapeutics · doi:10.1016/j.pharmthera.2016.07.010 · Cited by 107 · see citing works
“Echinomycin is an antibiotic that can bind DNA in a sequence-speci fi c fashion and that interferes with HIF-1-induced gene transcription by binding to its hypoxia response elements (Kong et al., 2005).”
Hatano, N., Itoh, Y., Suzuki, H., Muraki, Y., Hayashi, H., Onozaki, K., Wood, I. C., Beech, D. J., & Muraki, K. (2012) · Journal of Biological Chemistry · doi:10.1074/jbc.m112.361139 · Cited by 108 · see citing works
“over, Echi and YC-1, which reduceHIF1 activity as HIF inhibitors (32, 33), effectively inhibited the action of TNF .”
BackgroundZhang, J., Hu, Z., Horta, C. A., & Yang, J. (2022) · Seminars in Cancer Biology · doi:10.1016/j.semcancer.2022.12.002 · Cited by 103 · see citing works
Dynamic regulation of stem cell specification and maintenance by hypoxia-inducible factors
Semenza, G. L. (2015) · Molecular Aspects of Medicine · doi:10.1016/j.mam.2015.09.004 · Cited by 92 · see citing works
Hemin-induced necroptosis involves glutathione depletion in mouse astrocytes.
Laird, M. D., Wakade, C., Alleyne, C. H., & Dhandapani, K. (2008) · Free Radical Biology & Medicine · doi:10.1016/j.freeradbiomed.2008.07.003 · Cited by 108 · see citing works
Kwon, T., Zhao, X., Yang, Q., Li, Y., Ge, C., Zhao, G., & Franceschi, R. T. (2011) · Journal of Cellular Biochemistry · doi:10.1002/jcb.23289 · Cited by 102 · see citing works
Jin, W., Zhao, J., Yang, E., Wang, Y., Wang, Q., Wu, Y., Tong, F., Tan, Y., Zhou, J., & Kang, C. (2022) · Theranostics · doi:10.7150/thno.71029 · Cited by 104 · see citing works
“Echinomycin is a potent small-molecule and cell-permeable inhibitor of HIF-1α DNA-binding activity [34].”
BackgroundAML1/ETO cooperates with HIF1α to promote leukemogenesis through DNMT3a transactivation
Gao, X., Yan, F., Lin, J., Gao, L., Lu, X., Wei, S., Shen, N., Pang, J. X., Ning, Q., Komeno, Y., et al. (2015) · Leukemia · doi:10.1038/leu.2015.56 · Cited by 91 · see citing works
- Authors
- Kong, D., Park, E. J., Stephen, A. G., Calvani, M., Cardellina, J. H., Monks, A., Fisher, R. J., Shoemaker, R. H., & Melillo, G.
- Affiliations
- Science Applications International Corporation (United States) (US)
- National Cancer Institute (US)
- Venue
- Cancer Research
- Published
- 2005-10-01
- Volume
- 65
- Issue
- 19
- Pages
- 9047-9055
- ISSN
- 0008-5472 (ISSN-L)
- 1538-7445
- OpenAlex
- W2144670533
- Cited by
- 508
- References
- 53
- Citations by year
- 2026: 12 · 2025: 24 · 2024: 23 · 2023: 25 · 2022: 31 · 2021: 34 · 2020: 26 · 2019: 22 · 2018: 23 · 2017: 26 · 2016: 22 · 2015: 23 · 2014: 20 · 2013: 23 · 2012: 26
- FWCI
- 7.64
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Cancer Research (publishedVersion)
- PubMed (publishedVersion)
- Florence Research (University of Florence) (submittedVersion, other-oa)
- Funders
- National Institutes of Health (n01co12400-79-0-22, N01-CO-12400)
- National Cancer Institute (N01CO12400)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Physical Sciences › Chemistry › Organic Chemistry › Synthesis and biological activity
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Kruppel-like factors research
- Keywords
- echinomycin, HIF-1, hypoxia response element, transcription factor-DNA binding, small molecule inhibitors, VEGF promoter, angiogenesis, tumor progression, metastasis, high-throughput screening
- MeSH terms
- Antibiotics, Antineoplastic, Breast Neoplasms, DNA, Neoplasm, Echinomycin, Enzyme-Linked Immunosorbent Assay, Glioma, Humans, Luciferases, Promoter Regions, Genetic, Recombinant Proteins, RNA, Messenger, Vascular Endothelial Growth Factor A, Cell Line, Tumor, Hypoxia-Inducible Factor 1
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-11.
Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-11.
Cited sources are checked by gpt-6-sol.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
