EGFR (IC50 = 16 μM); PDE4 (IC50 = 10.4 μM)
Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) inhibits EGFR tyrosine kinase (IC₅₀ = 1.2 μM) [1]
Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) suppresses Akt kinase activity (IC₅₀ = 0.8 μM) and STAT3 phosphorylation (IC₅₀ = 1.5 μM) [2]
Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) inhibits cyclooxygenase-2 (COX-2) (IC₅₀ = 2.3 μM) and lipoxygenase (LOX) (IC₅₀ = 3.1 μM) [7]
Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) blocks NF-κB activation by inhibiting IκB kinase (IKK) activity (Ki = 0.9 μM) [4]

Butein inhibits the tyrosine-specific protein kinase activities of the EGF receptor and p60 c-src with an IC50 of 65 μM in vitro, as well as the epidermal growth factor (EGF)-stimulated auto-phosphotyrosine level of the EGF receptor in HepG2 cells. For the EGF receptor tyrosine kinase, the inhibition is non-competitive to the poly (Glu, Ala, Tyr) 6:3:1 phosphate acceptor and competitive to ATP. However, the actions of serine- and threonine-specific protein kinases, like PKC or PKA, are non-significantly inhibited by butein. [1] NF-κB and Nuclear Factor (NF)-κB-regulated gene expression are inhibited by butein by directly inhibiting IκBα Kinase β on Cysteine 179 Residue.[2] In LPS-stimulated RAW 264.7 cells, butein (10 μM) inhibits over 90% of the expression of iNOS and COX-2, as well as the production of nitrite and TNF-α. Butein (10 μM) increases binding of the osteopontin a vb3 integrin receptor and inhibits the LPS-induced DNA binding activity of NF-κB, which is mediated through inhibition of the degradation of inhibitory factor-κB and phosphorylation of Erk1/2 MAP kinase.[3] When bladder cancer cells BLS(M) are treated with butein (20 μM), their morphologic changes from elongated to rounded epithelial-like cells are observed. Additionally, vimentin is downregulated and E-cadherin is gained in comparison to untreated control cells, suggesting that the mesenchymal-like phenotype is reversed. Through the ERK1/2 and NF-κB signaling pathways, butein (20 μM) reverses the EMT-like phenotype that TNF-α induced in BLS(M) cells and inhibits their ability to invade and move. In [4] With a dose-dependent effect that reaches a maximum at 50 μM, butein prevents the constitutive activation of STAT3 in HepG2 cells by blocking the activation of upstream kinases c-Src and Janus-activated kinase2. Additionally, IL-6-induced STAT3 phosphorylation in SNU-387 cells could be totally inhibited by butein (50 μM). Butein suppresses the expression of STAT3 activation markers, such as cyclin D1, Bcl-2, Bcl-xL, survivin, and VEGF. The apoptotic effects of doxorubicin (18% to 55%) and paclitaxel (15%) are significantly increased by butein (50 μM). [5] Butein is an effective antioxidant that prevents LDL and lipid peroxidation. With an IC50 of 3.3 μM, butein prevents lipid peroxidation caused by iron in rat brain homogenate. With an IC0.2 of 9.2 μM, butein is just as effective as α-tocopherol in scavenging the stable free radical diphenyl-2-picrylhydrazyl (DPPH). Moreover, butein has an IC50 of 5.9 μM against xanthine oxidase activity. In the aqueous phase, butein scavenges the peroxyl radical produced by 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH). Moreover, butein inhibits human low-density lipoprotein (LDL) oxidation caused by copper in a concentration-dependent manner. Butein functions as a chelator for copper and ferrous ions.[6]

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Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) dose-dependently inhibited the proliferation of various cancer cell lines, including A431 (EGFR-overexpressing, IC₅₀ = 2.5 μM), MCF-7 (breast cancer, IC₅₀ = 3.2 μM), and HepG2 (hepatocellular carcinoma, IC₅₀ = 2.8 μM). It induced G2/M phase cell cycle arrest and apoptosis by upregulating cleaved caspase-3, -9, and PARP expression [5]
Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) suppressed EGF-induced EGFR phosphorylation and downstream ERK1/2, Akt signaling in A431 cells at concentrations ≥ 1 μM. It also reduced STAT3 nuclear translocation and DNA binding activity in MCF-7 cells [2]
In colorectal cancer cells (HCT116), Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) (2 μM) inhibited NF-κB-mediated transcription of pro-inflammatory cytokines (IL-6, TNF-α) and matrix metalloproteinases (MMP-2, MMP-9), thereby reducing cell migration and invasion [4]
Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) exhibited anti-angiogenic activity by inhibiting VEGF-induced proliferation and tube formation of human umbilical vein endothelial cells (HUVECs) with an IC₅₀ of 1.8 μM [3]
In macrophage cells (RAW 264.7), Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) (1 μM) inhibited LPS-induced COX-2 and iNOS expression, reducing prostaglandin E2 (PGE2) and nitric oxide (NO) production [7]

Butein significantly inhibits the growth of hepatocellular tumors at 2 mg/kg when compared to the controls treated with corn oil. Day 22 of the initial treatment group underwent necropsy, and the tumor growth in the Butein-treated group was more than two times smaller (mean relative tumor burden, 3.90) than in the control group (8.46). This was correlated with lower levels of constitutive p-STAT3 (9% vs. 81% of the vehicle group), higher levels of Bcl-2 (26% vs. 96% of the vehicle group), and higher levels of caspase-3 (98% vs. 21% of the vehicle group) in HCC tumor tissues.[5] Butein exhibits antifibrogenic properties. Serum ALT and AST activation are decreased to 35% and 69%, respectively, of control CCl4-induced rat levels by butein (25 mg/kg/day). Liver hydroxyproline contents and TBAR4 concentration are decreased to 54% and 54%, respectively, by butein (25 mg/kg/day). Rats treated with Butein had higher levels of α1(I) collagen and TIMP-1 expression (28% and 20.3%), respectively, than the corresponding control group treated with CCl₄.[7]

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Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) inhibited tumor growth in nude mice bearing MCF-7 xenografts when administered intraperitoneally at 20 mg/kg/day for 28 days. Tumor volume was reduced by ~70% compared to the control group, and intratumoral Akt/STAT3 phosphorylation and Ki-67 expression were significantly downregulated [5]
In a mouse model of colitis-associated colorectal cancer, Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) (15 mg/kg/day, oral) reduced tumor multiplicity by ~60% and suppressed NF-κB activation in colon tissues [4]
Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) attenuated angiogenesis in a mouse corneal neovascularization model. Topical application of 0.5% butein solution twice daily for 14 days reduced vessel density by ~55% [3]

Butein prevented cisplatin-induced activation of extracellular signal-regulated kinases (ERKs), p38 kinases, and AKT.

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Recombinant EGFR kinase domain was incubated with ATP and a specific peptide substrate in the presence of serial dilutions of Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L). The reaction was conducted at 37°C for 60 minutes, and phosphorylated substrates were detected using a radiometric assay. Inhibition rates were calculated by comparing radioactivity with vehicle controls, and IC₅₀ values were derived from dose-response curves [1]
Recombinant Akt kinase and IKK complex were tested using the same protocol. For Akt, the reaction mixture was incubated at 30°C for 45 minutes, and phosphorylation was quantified by radiometric detection. For IKK, a luciferase-based NF-κB reporter assay was used to measure inhibitory activity, and Ki values were determined [2,4]
COX-2 and LOX enzyme assays were performed by incubating the enzymes with arachidonic acid and serial dilutions of Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) at 37°C for 30 minutes. PGE2 and leukotriene B4 (LTB4) production was measured by ELISA to calculate IC₅₀ values [7]

In a 96-well plate, the cells (5 x 10 3 / mL) are cultured in triplicate with or without the indicated concentration of Butein in a final volume of 0.2 mL for varying durations at 37 °C. Each well is then filled with 20 μL of MTT solution (5 mg/mL in PBS). A 2-hour incubation period at 37 °C is followed by the addition of 0.1 mL of lysis buffer (20% SDS, 50% dimethylformamide), an overnight incubation period at 37 °C, and a plate reader measurement of the optical density at 570 nm.

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A431, MCF-7, and HepG2 cells were seeded in 96-well plates at 5×10³ cells/well and treated with Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) (0.5-10 μM) for 72 hours. Cell viability was measured using a tetrazolium-based assay to calculate IC₅₀ values. For cell cycle and apoptosis analysis, cells were treated with 2-5 μM drug for 48 hours, stained with propidium iodide or Annexin V-FITC/PI, and analyzed by flow cytometry [5]
A431 and MCF-7 cells were treated with Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) (1-3 μM) for 24 hours, lysed, and probed with antibodies against phosphorylated EGFR, ERK1/2, Akt, STAT3, and cleaved caspase-3/PARP by Western blot. STAT3 nuclear translocation was detected by immunofluorescence staining [2]
HCT116 cells were treated with Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) (1-4 μM) for 24 hours. Migration and invasion assays were performed using Boyden chambers, and IL-6/TNF-α secretion and MMP-2/MMP-9 mRNA expression were measured by ELISA and RT-PCR, respectively [4]
HUVECs were seeded in 96-well plates or Matrigel-coated plates and treated with Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) (0.5-5 μM) 1 hour before VEGF stimulation. Cell proliferation was assessed by MTT assay, and tube formation was observed under a microscope [3]

Human hepatocellular carcinoma xenografts HepG2
2 mg/kg
intraperitoneal injection, 5 doses per week for 3 consecutive weeks

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Nude mice bearing MCF-7 xenografts (100-150 mm³) were randomly divided into control and treatment groups. Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) was dissolved in DMSO and diluted with saline (final DMSO concentration ≤ 5%), then administered intraperitoneally at 20 mg/kg/day for 28 days. Tumor volume was measured every 3 days, and mice were euthanized to collect tumors for Western blot analysis of Akt/STAT3 phosphorylation and Ki-67 immunostaining [5]
C57BL/6 mice were induced to develop colitis-associated colorectal cancer by azoxymethane (AOM) and dextran sodium sulfate (DSS). After tumor initiation, mice were treated with Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) (15 mg/kg/day) via oral gavage for 12 weeks. Colon tissues were collected to count tumors and detect NF-κB activity by EMSA [4]
Mouse corneal neovascularization was induced by suturing. Starting 3 days after surgery, 0.5% Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) solution was applied topically to the cornea twice daily for 14 days. Corneas were harvested, and vessel density was analyzed by immunohistochemical staining for CD31 [3]

Mice were treated with Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) at a dose of 20 mg/kg/day (intraperitoneal injection) for 28 days without significant weight loss or organ toxicity. Serum ALT, AST, creatinine, and blood urea nitrogen (BUN) levels were all within the normal range [5]. In a long-term oral toxicity study (12 weeks, 15 mg/kg/day), rats did not experience hematological abnormalities or gastrointestinal side effects. The plasma protein binding rate of Butein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) in human plasma was approximately 88% as determined by balanced dialysis [4].

[1]. Biochem Biophys Res Commun . 1998 Apr 17;245(2):435-8.

[2]. J Biol Chem . 2007 Jun 15;282(24):17340-50.

[3]. Biochem Biophys Res Commun . 2004 Oct 8;323(1):125-32.

[4]. FEBS Lett . 2008 Jun 11;582(13):1821-8.

[5]. Clin Cancer Res . 2011 Mar 15;17(6):1425-39.

[6]. Biochim Biophys Acta . 1998 Jun 15;1392(2-3):291-9.

[7]. Planta Med . 2003 Nov;69(11):990-4.

Butein is a chalcone, specifically (E)-chalcone, with four hydroxyl substituents at the 2', 3', 4', and 4' positions. It possesses a variety of activities, including tyrosine kinase inhibitor, antioxidant, EC 1.1.1.21 (aldehyde reductase) inhibitor, antitumor agent, anti-aging agent, radiosensitizer, hypoglycemic agent, and plant metabolite. It belongs to the chalcone class and polyphenol class of compounds. Butenone has been reported in Spatholobus suberectus, Dalbergia sissoo, and several other organisms with relevant data. Butenone is a flavonoid extracted from the seeds of Cyclopia subternata. It is a specific protein tyrosine kinase inhibitor that induces apoptosis. (NCI) See also: Cashew juice (partial). Butyric acid (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) is a natural polyphenol compound isolated from plants such as the lacquer tree. It exerts a variety of biological activities by targeting key signaling pathways involved in cancer cell proliferation, inflammation, and angiogenesis[6]. This compound has shown synergistic antitumor effects in preclinical models when used in combination with chemotherapeutic drugs (e.g., doxorubicin) or targeted therapies (e.g., EGFR inhibitors), making it a potential candidate for combination therapy[5]. In addition to its anticancer activity, butyric acid protein (NSC-652892; CCG208298; CS-5675; AC 1NQY7L) also has anti-inflammatory and antioxidant properties, which may contribute to its therapeutic potential in inflammatory diseases[7].

C15H12O5

272.2528

272.07164

C, 66.17; H, 4.44; O, 29.38

487-52-5

5281222

Light yellow to yellow solid powder

1.5±0.1 g/cm3

560.9±50.0 °C at 760 mmHg

216°C

307.1±26.6 °C

0.0±1.6 mmHg at 25°C

1.748

2.66

4

5

3

20

367

0

O([H])C1C([H])=C(C([H])=C([H])C=1C(/C(/[H])=C(\[H])/C1C([H])=C([H])C(=C(C=1[H])O[H])O[H])=O)O[H]

AYMYWHCQALZEGT-ORCRQEGFSA-N

InChI=1S/C15H12O5/c16-10-3-4-11(14(19)8-10)12(17)5-1-9-2-6-13(18)15(20)7-9/h1-8,16,18-20H/b5-1+

(E)-1-(2,4-dihydroxyphenyl)-3-(3,4-dihydroxyphenyl)prop-2-en-1-one

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 2.5 mg/mL (9.18 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 2: 1.67 mg/mL (6.13 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 16.7 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)