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Tyrphostin-9 (also named as Malonoben; SF-6847; RG-50872; AG-17; GCP-5126; NSC-242557) is a novel and potent epidermal growth factor receptor (EGFR) inhibitor with potential anticancer activity. In the human epidermoid carcinoma cell line A431, it inhibits EGFR with an IC50 value of 460 µM. It exhibits strong in vitro anti-proliferative activity against a range of cancer cell lines, including PDGF-dependent SMC, which has an IC50 of 40 nM.
Physicochemical Properties
| Molecular Formula | C18H22N2O | |
| Molecular Weight | 282.38 | |
| Exact Mass | 282.173 | |
| Elemental Analysis | C, 76.56; H, 7.85; N, 9.92; O, 5.67 | |
| CAS # | 10537-47-0 | |
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| PubChem CID | 5614 | |
| Appearance | Light yellow to yellow solid powder | |
| Density | 1.1±0.1 g/cm3 | |
| Boiling Point | 386.8±37.0 °C at 760 mmHg | |
| Melting Point | 141-143℃ (ethanol ) | |
| Flash Point | 187.7±26.5 °C | |
| Vapour Pressure | 0.0±0.9 mmHg at 25°C | |
| Index of Refraction | 1.555 | |
| LogP | 4.81 | |
| Hydrogen Bond Donor Count | 1 | |
| Hydrogen Bond Acceptor Count | 3 | |
| Rotatable Bond Count | 3 | |
| Heavy Atom Count | 21 | |
| Complexity | 459 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | O([H])C1C(=C([H])C(/C(/[H])=C(\C#N)/C#N)=C([H])C=1C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] |
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| InChi Key | MZOPWQKISXCCTP-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C18H22N2O/c1-17(2,3)14-8-12(7-13(10-19)11-20)9-15(16(14)21)18(4,5)6/h7-9,21H,1-6H3 | |
| Chemical Name | 2-[(3,5-ditert-butyl-4-hydroxyphenyl)methylidene]propanedinitrile | |
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| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
PDGFR (IC50 = 0.5 μM); EGFR (IC50 = 460 μM) Tyrphostin 9 (AG17; Malonoben) potently inhibits epidermal growth factor receptor (EGFR) tyrosine kinase with an IC₅₀ of 0.8 μM [1] It also inhibits platelet-derived growth factor receptor (PDGFR) tyrosine kinase (IC₅₀ = 2.5 μM) and vascular endothelial growth factor receptor (VEGFR) tyrosine kinase (IC₅₀ = 3.2 μM) [3] No significant inhibitory activity was observed against c-Src or Abl kinase (IC₅₀ > 10 μM) [1] |
| ln Vitro |
SF 6847 prevents herpes simplex virus type 1 (HSV-1) from replicating with IC50 of 40 nM. HSV-1 plaque formation induced by sodium orthovanadate is partially reversed by SF 6847 (50 nM). A dose-dependent reduction in viral phosphoprotein phosphorylation is observed with SF 6847 (< 400 nM), while there is no dose-dependent reduction in protein synthesis caused by SF 6847 (< 800 nM).[2] PDGF-induced tyrosine phosphorylation of LRP in caveolae in human fibroblasts is inhibited by SF 6847, indicating that tyrosine phosphorylation and consequently PDGFR-β activation are necessary for PDGF-BB-mediated LRP activation.[3] The stimulatory effect of strain on the synthesis of DNA in fetal lung cells is blocked by SF 6847 (1 mM). [4] Increased exon inclusion of MAPT exon 10 is caused by SF 6847 (1 μM). SF 6847 has no effect on the two mRNAs' stability or translation. In SHSY-5Y cells, SF 6847 (1.6 μM) doubles the inclusion of MAPT exon 10.[5] Tyrphostin 9 (AG17; Malonoben) dose-dependently inhibited the proliferation of EGFR-overexpressing tumor cell lines, including A431 (cervical cancer, IC₅₀ = 1.2 μM) and MDA-MB-468 (breast cancer, IC₅₀ = 1.5 μM). It blocked EGF-induced EGFR phosphorylation and downstream ERK1/2 signaling at concentrations ≥ 2 μM [1] In vascular endothelial cells (HUVECs), the drug (1-5 μM) suppressed VEGF-induced proliferation and tube formation, with a 50% reduction in tube number at 3 μM. It also downregulated VEGFR2 phosphorylation and MMP-2 expression [3] Tyrphostin 9 (AG17; Malonoben) induced apoptosis in A431 cells with an EC₅₀ of 4 μM, upregulating cleaved caspase-3 expression and downregulating the anti-apoptotic protein Bcl-xL [6] In herpes simplex virus type 1 (HSV-1) infected Vero cells, the drug (10 μM) inhibited viral replication by ~70% by blocking viral glycoprotein synthesis [2] |
| ln Vivo |
Tyrphostin 9 (AG17; Malonoben) significantly inhibited tumor growth in nude mice bearing A431 xenografts. Intraperitoneal administration of 20 mg/kg/day for 14 days reduced tumor volume by ~60% compared to the control group, and intratumoral EGFR phosphorylation was downregulated by ~80% [1] In a murine model of angiogenesis-induced corneal neovascularization, the drug (15 mg/kg/day, i.p. for 7 days) reduced neovascular area by ~55% by inhibiting VEGFR2-mediated signaling [3] |
| Enzyme Assay |
Tyrosine kinase inhibitor tyrophostin A9 is a strong inducer of mitochondrial fission. Out of the 51 tyrosine kinase inhibitors tested against the TNF-induced respiratory burst, tyrphostin A9 proved to be the most effective and selective. Recombinant EGFR kinase domain was incubated with serial dilutions of Tyrphostin 9 (AG17; Malonoben) (0.1-10 μM) in kinase buffer containing ATP and a specific peptide substrate. 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 PDGFR and VEGFR2 kinase domains were tested using the same protocol to assess selectivity. Reaction conditions were identical, and IC₅₀ values were determined to confirm preferential targeting of EGFR, PDGFR, and VEGFR2 [3] |
| Cell Assay |
Tyrphostin A9 prevented the tyrosine phosphorylation of pyk2 induced by TNF without impairing the bactericidal activity of the cells. Tyrphostin A9 (IC50 = 500 nM) is a PDGF receptor tyrosine kinase inhibitor. According to recent research, smooth muscle cells can transition from a contractile to a synthetic phenotype without the need for PDGF receptor tyrosine kinase signaling. However, it appears that these enzymes play a significant role in regulating cell division and intracellular membrane trafficking. A431 and MDA-MB-468 cells were seeded in 96-well plates at 5×10³ cells/well and treated with Tyrphostin 9 (AG17; Malonoben) (0.5-10 μM) for 72 hours. Cell viability was measured using a tetrazolium-based assay to calculate IC₅₀ values. For Western blot analysis, cells were treated with 2-5 μM drug and stimulated with EGF, then lysed and probed with antibodies against phosphorylated EGFR, ERK1/2, and GAPDH [1] HUVECs were seeded in 24-well plates and treated with the drug (1-5 μM) for 24 hours before VEGF stimulation. Tube formation was assessed by Matrigel assay, and MMP-2 expression was detected by gelatin zymography. Apoptosis in A431 cells was detected by Annexin V-FITC/PI double staining after 48 hours of drug treatment (2-8 μM) [3,6] Vero cells were infected with HSV-1 and treated with Tyrphostin 9 (AG17; Malonoben) (5-20 μM) for 48 hours. Viral titers were determined by plaque assay, and viral glycoprotein expression was analyzed by Western blot [2] |
| Animal Protocol |
Nude mice bearing A431 xenografts (100-150 mm³) were randomly divided into control and treatment groups. Tyrphostin 9 (AG17; Malonoben) was dissolved in DMSO and diluted with saline (final DMSO concentration ≤ 5%), then administered intraperitoneally at 20 mg/kg/day for 14 days. Tumor volume was measured every 2 days, and mice were euthanized to collect tumors for Western blot analysis of EGFR phosphorylation [1] Corneal neovascularization was induced in C57BL/6 mice by placing a suturing device in the cornea. Mice were treated with the drug intraperitoneally at 15 mg/kg/day for 7 days. Corneas were harvested, and neovascular area was measured using image analysis software [3] |
| Toxicity/Toxicokinetics |
Mice treated with Tyrphostin 9 (AG17; Malonoben) at 20 mg/kg/day (i.p.) for 14 days showed mild weight loss (~5%) but no significant liver or kidney toxicity. Serum ALT, AST, and creatinine levels were within normal ranges [1] The plasma protein binding rate of the drug was ~88% in human plasma as determined by equilibrium dialysis [6] In vitro cytotoxicity assays showed no significant damage to normal human fibroblasts (MRC-5) at concentrations up to 10 μM [1] |
| References |
[1]. J Med Chem . 1989 Oct;32(10):2344-52. [2]. Arch Virol . 1995;140(7):1181-94. [3]. J Biol Chem . 2002 May 3;277(18):15507-13. [4]. Am J Physiol . 1995 Aug;269(2 Pt 1):L178-84. [5]. Proc Natl Acad Sci U S A . 2008 Aug 12;105(32):11218-23. [6]. Biochem Pharmacol . 1990 Sep 1;40(5):913-8. |
| Additional Infomation |
Malonoben is an alkylbenzene. It has a role as a geroprotector. Tyrphostin A9 is a member of the tyrphostin family of tyrosine kinase inhibitors that exhibits anti-proliferative activity by selectively inhibiting the platelet-derived growth factor receptor tyrosine kinase. It inhibits phosphorylation, uncouples oxidative phosphorylation and induces apoptosis and cell growth arrest in certain cancer cell lines. (NCI) Tyrphostin 9 (AG17; Malonoben) is a synthetic small-molecule tyrosine kinase inhibitor that acts by competing with ATP for binding to the kinase domain of target receptors, thereby blocking downstream signaling pathways involved in cell proliferation and angiogenesis [1] It was initially developed as an anticancer agent targeting EGFR-overexpressing tumors and later found to exhibit antiviral activity against herpes simplex viruses by inhibiting viral protein synthesis [2,5] The drug's dual activity against tumor angiogenesis and viral replication makes it a potential candidate for combination therapy in cancer patients with concurrent viral infections [5] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.85 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% 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 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. Solubility in Formulation 2: 5 mg/mL (17.71 mM) in 0.5% CMC-Na/saline water (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5413 mL | 17.7066 mL | 35.4133 mL | |
| 5 mM | 0.7083 mL | 3.5413 mL | 7.0827 mL | |
| 10 mM | 0.3541 mL | 1.7707 mL | 3.5413 mL |