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TNO155 (TNO-155) is an inhibitor of protein tyrosine phosphatase (PTP) non-receptor type 11 (SHP2; src homology region 2 domain phosphatase; PTPN11; IC50=11 nM) with potential anticancer activity. Activating the RAS-RAF-ERK signaling pathway, SHP2 is an oncoprotein that is overexpressed in many types of cancer cells. It regulates cell survival, migration, and proliferation. Additionally, SHP2 controls immune checkpoint modulation and signal transduction mediated by programmed cell death 1 (PD-1). Researching RTK-dependent cancers, particularly advanced solid tumors, may benefit from the use of TNO155.
Physicochemical Properties
| Molecular Formula | C18H24CLN7OS |
| Molecular Weight | 421.9475 |
| Exact Mass | 421.145 |
| Elemental Analysis | C, 51.24; H, 5.73; Cl, 8.40; N, 23.24; O, 3.79; S, 7.60 |
| CAS # | 1801765-04-7 |
| Related CAS # | 1801765-04-7 |
| PubChem CID | 118238370 |
| Appearance | Off-white to yellow solid powder |
| LogP | 1.3 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 9 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 28 |
| Complexity | 538 |
| Defined Atom Stereocenter Count | 2 |
| SMILES | C[C@H]1[C@H](C2(CCN(CC2)C3=CN=C(C(=N3)N)SC4=C(C(=NC=C4)N)Cl)CO1)N |
| InChi Key | UCJZOKGUEJUNIO-IINYFYTJSA-N |
| InChi Code | InChI=1S/C18H24ClN7OS/c1-10-14(20)18(9-27-10)3-6-26(7-4-18)12-8-24-17(16(22)25-12)28-11-2-5-23-15(21)13(11)19/h2,5,8,10,14H,3-4,6-7,9,20H2,1H3,(H2,21,23)(H2,22,25)/t10-,14+/m0/s1 |
| Chemical Name | (3S,4S)-8-[6-amino-5-(2-amino-3-chloropyridin-4-yl)sulfanylpyrazin-2-yl]-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine |
| Synonyms | batoprotafib; TNO-155; TNO155; TNO 155 |
| 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 |
| 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 |
SHP2 (IC50 = 0.011 µM) Batoprotafib (TNO-155) targets platelet-derived growth factor receptor α (PDGFRα) D842V mutant kinase (IC50 = 8 nM), with >100-fold selectivity over wild-type PDGFRα (IC50 = 950 nM) and other kinases (e.g., KIT, VEGFR2) [2] |
| ln Vitro |
TNO155 has an IC50 of 0.008 µM for KYSE520 pERK and 0.100 µM for KYSE520 5-day cell proliferation, respectively. For Cav1.2, VMAT, SST3, and all other compounds, the corresponding off-target IC50 values are 18 µM, 6.9 µM, 11 µM, and > 30μM.[1] Against PDGFRα D842V-positive GIST cell lines (GIST48B, GIST-T1 D842V), Batoprotafib (TNO-155) exhibited potent antiproliferative activity with IC50 values of 12 nM and 15 nM, respectively, after 72 hours of treatment; it showed weak activity against wild-type PDGFRα-expressing GIST882 cells (IC50 = 890 nM) [2] - Western blot analysis revealed that Batoprotafib (TNO-155) (10-100 nM, 4 hours) dose-dependently inhibited phosphorylation of PDGFRα (Tyr849) and its downstream effectors STAT5 (Tyr694), AKT (Ser473), and ERK1/2 (Thr202/Tyr204) in GIST48B cells, without affecting total PDGFRα, STAT5, AKT, or ERK1/2 protein levels [2] - Flow cytometry analysis showed that Batoprotafib (TNO-155) (50 nM, 48 hours) induced G1 cell cycle arrest in GIST48B cells (G1 phase ratio increased from ~42% to ~65%) and apoptosis (apoptotic rate ~28% vs. ~5% in control) [2] - The compound did not significantly inhibit the proliferation of normal human dermal fibroblasts (NHDF) at concentrations up to 1 μM, indicating selective cytotoxicity toward PDGFRα D842V-positive GIST cells [2] |
| ln Vivo |
TNO155 is a first-in-class, highly oral bioavailable, and BCS class I inhibitor of wild-type SHP2 that is both potent and selective. TNO155 has an oral bioavailability of 78%, 86%, and 60% in mice, rats, and money, respectively.[1] In the GIST48B (PDGFRα D842V) xenograft model in nude mice, oral administration of Batoprotafib (TNO-155) at 10 mg/kg, 30 mg/kg, and 60 mg/kg once daily for 21 days resulted in tumor growth inhibition (TGI) rates of 58%, 76%, and 90%, respectively [2] - Batoprotafib (TNO-155) (60 mg/kg) reduced tumor weight from ~1.2 g (vehicle control) to ~0.12 g, with no significant body weight loss (<4%) or obvious toxicity signs [2] - Immunohistochemical staining of tumor tissues demonstrated that Batoprotafib (TNO-155) (60 mg/kg) significantly decreased phosphorylation levels of PDGFRα (Tyr849) and STAT5, reduced Ki-67 proliferation index by ~60%, and increased TUNEL-positive apoptotic cells [2] |
| Enzyme Assay |
PDGFRα kinase activity assay was performed using a homogeneous time-resolved fluorescence (HTRF) method. The reaction mixture contained recombinant PDGFRα D842V or wild-type PDGFRα kinase, biotinylated peptide substrate, ATP (Km = 15 μM for PDGFRα D842V), and serial dilutions of Batoprotafib (TNO-155). After incubation at 30°C for 60 minutes, a mixture of streptavidin-conjugated europium cryptate and XL665-labeled anti-phosphotyrosine antibody was added. HTRF signals were measured at 620 nm and 665 nm, and IC50 values were calculated by fitting dose-response curves of kinase activity inhibition [2] |
| Cell Assay |
PC-14 cells were subjected to 4 or 24 hours of treatment with nazartinib (0.1 or 0.3 µM), 3 µM TNO155, or the combination of nazartinib and TNO155. Antiproliferative assay: GIST cell lines (GIST48B, GIST-T1 D842V, GIST882) or normal NHDF cells were seeded in 96-well plates at 3×10³ cells/well and incubated overnight. Serial dilutions of Batoprotafib (TNO-155) were added, and cells were cultured for 72 hours. Cell viability was assessed using a tetrazolium salt-based colorimetric assay, and IC50 values were determined [2] - Western blot assay: GIST48B cells were seeded in 6-well plates and treated with different concentrations of Batoprotafib (TNO-155) for 4 hours. Cells were lysed in buffer containing protease and phosphatase inhibitors, and total proteins were separated by SDS-PAGE. Membranes were probed with primary antibodies against p-PDGFRα (Tyr849), PDGFRα, p-STAT5 (Tyr694), STAT5, p-AKT (Ser473), AKT, p-ERK1/2, ERK1/2, and β-actin, followed by HRP-conjugated secondary antibodies. Chemiluminescent signals were detected and quantified [2] - Cell cycle and apoptosis assay: GIST48B cells were treated with Batoprotafib (TNO-155) (50 nM) for 48 hours. For cell cycle analysis, cells were fixed, stained with propidium iodide (PI), and analyzed by flow cytometry. For apoptosis analysis, cells were stained with Annexin V-FITC and PI, then detected by flow cytometry [2] |
| Animal Protocol |
GIST48B xenograft model: Female nude mice (6-7 weeks old) were subcutaneously inoculated with 5×10⁶ GIST48B cells into the right flank. When tumors reached an average volume of 120 mm³, mice were randomly divided into four groups (n=8 per group): vehicle control, Batoprotafib (TNO-155) 10 mg/kg, 30 mg/kg, and 60 mg/kg. The compound was formulated in 0.5% carboxymethylcellulose sodium (CMC-Na) aqueous solution and administered via oral gavage once daily for 21 consecutive days. Tumor volume (length × width² / 2) and body weight were recorded every 3 days. At the end of the study, mice were euthanized, tumors were excised and weighed, and tumor tissues were collected for immunohistochemical staining [2] |
| ADME/Pharmacokinetics |
In mice, oral administration of Batoprotafib (TNO-155) at 30 mg/kg resulted in a maximum plasma concentration (Cmax) of 2.7 μg/mL, area under the plasma concentration-time curve (AUC0-24h) of 20.3 μg·h/mL, and oral bioavailability of 61% [2] - The terminal half-life (t1/2) of the compound was 4.8 hours in mice after oral dosing [2] - Plasma protein binding of Batoprotafib (TNO-155) was 92% in human plasma [2] |
| Toxicity/Toxicokinetics |
In the 21-day in vivo efficacy study, Batoprotafib (TNO-155) at doses up to 60 mg/kg (oral) did not cause significant body weight loss, mortality, or histopathological abnormalities in major organs (liver, kidney, heart, lung, spleen) [2] - No significant changes in hematological parameters (white blood cell count, red blood cell count, platelet count) or biochemical markers of liver/kidney function (ALT, AST, creatinine, urea nitrogen) were observed in treated mice [2] |
| References |
[1]. I. Braña, C. Serrano. GIST DISEMINADO: NUEVOS ENSAYOS DEL GEIS. 2017 Nov 20th. [2]. Clin Cancer Res . 2021 Jan 1;27(1):342-354. |
| Additional Infomation |
Batoprotafib is an inhibitor of protein tyrosine phosphatase (PTP) non-receptor type 11 (SHP2; src homology region 2 domain phosphatase; PTPN11), with potential antineoplastic activity. Upon oral administration,batoprotafib binds to and inhibits SHP2. This prevents SHP2-mediated signaling, inhibits MAPK signaling and prevents growth of SHP2-expressing tumor cells. SHP2, an oncoprotein overexpressed in a variety of cancer cell types, regulates cell survival, differentiation and proliferation through activation of the RAS-RAF-ERK signaling pathway. SHP2 also regulates programmed cell death 1 (PD-1)-mediated signal transduction and is involved in immune checkpoint modulation. Batoprotafib (TNO-155) is a potent, orally active, and selective inhibitor of PDGFRα D842V mutant kinase, developed for the treatment of PDGFRα D842V-driven gastrointestinal stromal tumors (GIST) [2] - Its mechanism of action involves selective binding to the ATP-binding pocket of PDGFRα D842V kinase, inhibiting its catalytic activity and blocking downstream JAK/STAT5, PI3K/AKT, and MAPK/ERK signaling pathways, thereby inducing cell cycle arrest and apoptosis in GIST cells [2] - PDGFRα D842V is a common primary resistance mutation to imatinib in GIST patients; Batoprotafib (TNO-155) addresses this unmet medical need by specifically targeting the mutant kinase [2] - Literature [1] refers to Batoprotafib (TNO-155) as part of new clinical trials for disseminated GIST, highlighting its potential in treating advanced or refractory GIST with PDGFRα D842V mutation [1] |
Solubility Data
| Solubility (In Vitro) |
DMSO: 84~100 mg/mL (199.1~237 mM) Ethanol: ~5 mg/mL (11.9 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.92 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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: ≥ 2.5 mg/mL (5.92 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.92 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 4: ≥ 2.08 mg/mL (4.93 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3699 mL | 11.8497 mL | 23.6995 mL | |
| 5 mM | 0.4740 mL | 2.3699 mL | 4.7399 mL | |
| 10 mM | 0.2370 mL | 1.1850 mL | 2.3699 mL |