S3I-201

Catalog No.S1155 Synonyms: NSC 74859

For research use only.

S3I-201 (NSC 74859) shows potent inhibition of STAT3 DNA-binding activity with IC50 of 86 μM in cell-free assays, and low activity towards STAT1 and STAT5.

S3I-201 Chemical Structure

CAS No. 501919-59-1

Selleck's S3I-201 has been cited by 183 publications

Purity & Quality Control

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Biological Activity

Description S3I-201 (NSC 74859) shows potent inhibition of STAT3 DNA-binding activity with IC50 of 86 μM in cell-free assays, and low activity towards STAT1 and STAT5.
Features A chemical probe inhibitor of Stat3 activity.
Targets
STAT3 [1]
(Cell-free assay)
86 μM
In vitro

S3I-201 inhibits growth and induces apoptosis preferentially in tumor cells that contain persistently activated Stat3 by inhibiting Stat3·Stat3 complex formation and Stat3 DNA-binding and transcriptional activitie. Moreover, S3I-201 also inhibits the expression of the Stat3-regulated genes encoding cyclin D1, Bcl-xL, and survivin. [1] S3I-201 inhibits breast carcinoma MDA-MB-435, MDA-MB-453 and MDA-MB-231 cell lines with IC50 of 100 μM. In addition, the cells with impaired TGF-β signaling are four times as sensitive to the STAT3 inhibitor S3I-201. [2] A recent study shows that S3I-201 potentiates the antiproliferative effect of cetuximab in HepG2 and Huh-7 cells via the STAT3 signalling pathway. [3]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
U87 Mly3S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MYm3NkBp MkLlTWM2OD13NT6xJO69VQ>? NHzwfIE9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MEC3NlY2Oid-MkCwO|I3PTJ:L3G+
U373  MnOxS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NYDl[|NxPzJiaB?= MoLnTWM2OD13Mj61JO69VQ>? MlfJQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjByN{K2OVIoRjJyMEeyOlUzRC:jPh?=
HPAC MmCyS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M4L5clczKGh? M1XZfmlEPTExvK6xNFAh|ryP MV[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zODB5Mk[1Nkc,OjByN{K2OVI9N2F-
PANC-1 MlH5S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? Mmi3O|IhcA>? NEW0[3ZKSzVy78{eNVAxKM7:TR?= M4PacVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJyMEeyOlUzLz5{MEC3NlY2OjxxYU6=
SK-BR-3 Mnj0S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NH:xRlg4OiCq M{H4Z2lEPTExvK6xNFAh|ryP M4K3blxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJyMEeyOlUzLz5{MEC3NlY2OjxxYU6=
 U-373 MG M2LoT2N6fG:2b4jpZ4l1gSCDc4PhfS=> MYKzM|ExKM7:TR?= MmHmNlQhcA>? M17oRpJm\HWlZYOgSm4u|rNvaX7keYNm\CClZXzsJI5mfXKxdH;4bYNqfHl? NGXVdFU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MEi4PFQyPid-MkC4PFg1OTZ:L3G+
MDA-MB-231 MnjzS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MXm3NkBp MVLJR|Ux97zgMUCwJO69VQ>? M3LWSlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJyMEeyOlUzLz5{MEC3NlY2OjxxYU6=
HUVEC  NV2xV4w6TnWwY4Tpc44hSXO|YYm= NU\Zd5BoOC53LUKwJO69VQ>? M1mwZ|I1KGh? M4WwOGROW09? Mnv0d5VxeHKnc4Pld{B1cGViaInwc5hq[S2rbnT1Z4VlKGGlY4XteYxifGmxbjDv[kBJUUZvMd8x NW\TcWhbRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkG1NlM2PTlpPkKxOVI{PTV7PD;hQi=>
Huh7 NHLq[4dIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MkXrNVAxKG6P MlzSOFghcA>? MUTEUXNQ NXqydWEzcW6qaXLpeJMhfGinIFnMMVYhe3SrbYXsZZRqd25icILvcY91\WRiY3XscEBxem:uaX\ldoF1cW:w MoHpQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN|NkSzPFkoRjJ|M{[0N|g6RC:jPh?=
PLC/PRF/5  MXjHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M3nVNFExOCCwTR?= MXK0PEBp NYXzfXhXTE2VTx?= NFjzPJhqdmirYnn0d{B1cGViSVytOkB{fGmvdXzheIlwdiCycn;tc5Rm\CClZXzsJJBzd2yrZnXyZZRqd25? NIXndFU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{O2OFM5QSd-MkOzOlQ{QDl:L3G+
H460  NYTYZ5RrTnWwY4Tpc44hSXO|YYm= MXm1NE8yODBizszN MkfVOFghcA>? NYjK[od{cW6qaXLpeJMhfGinIGP0ZZQ{SyCrbnPy[YF{\WRibXnSMVkz[SCneIDy[ZN{cW:w MVe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzh{MEK1OEc,OjN6MkCyOVQ9N2F-
H1299 MU\GeY5kfGmxbjDBd5NigQ>? NIPmepM2OC9zMECg{txO MVi0PEBp NWfHSWlqe3WycILld5NmeyCvaWKtPVJiKGW6cILld5Nqd25iZH;z[U1l\XCnbnTlcpRtgQ>? MX:8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzh{MEK1OEc,OjN6MkCyOVQ9N2F-
T-cell  MXvHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MXfJR|UxRTVyIN88US=> MYK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDB4OEezNUc,OjRyNki3N|E9N2F-
U373  M4fiZmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MYKxNlUh|ryP M1LZbVI1KGh? NEHPNJhFVVOR MYHkbZNzfXC2czDTWGFVOyC|aXfuZYxqdmdiYX7kJJBzd2yrZnXyZZRqd25? MlXsQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjRyN{C4NlAoRjJ2MEewPFIxRC:jPh?=
HUT-102 NHXyR5dCeG:ydH;zbZMhSXO|YYm= NGO1[mQ4PS1|MECg{txO NHLOXoQzPC92ODDo M{PqXpN2eHC{ZYPz[ZMh[2WubDDwdo9tcW[ncnH0bY9vKGmwIHGg[I9{\S2mZYDlcoRmdnRibXHucoVzKGGwZDDpcoR2[2W|IHPlcIwh[XCxcITvd4l{yqB? NETRWnY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEC5NFk6PSd-MkSwPVA6QTV:L3G+
MT-2 NW[yXFlLSXCxcITvd4l{KEG|c3H5 MUC3OU0{ODBizszN Ml;oNlQwPDhiaB?= MYLzeZBxemW|c3XzJINmdGxicILvcIln\XKjdHnvckBqdiCjIHTvd4Uu\GWyZX7k[Y51KG2jbn7ldkBidmRiaX7keYNmeyClZXzsJIFxd3C2b4Ppd:Kh NXvIeFI1RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkSwPVA6QTVpPkK0NFkxQTl3PD;hQi=>
H460  NX;P[lhOSXCxcITvd4l{KEG|c3H5 M3\iUFExOMLibl2= NUPIXplZOjUEoHi= Mlv1[Y5p[W6lZYOgZ4VtdCCmZXH0bEBkdy22cnXheIVlKHerdHigUHkzQTRyMEK= NVi4TGVPRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS0O|I2OzhpPkK0OFczPTN6PD;hQi=>
A459 M2LScGFxd3C2b4Ppd{BCe3OjeR?= Mlv3NVAxyqCwTR?= NXKwVGJmOjUEoHi= M4GwU4lv\HWlZYOgZ4VtdCCjcH;weI9{cXNiY3:teJJm[XSnZDD3bZRpKEKHWkKzOS=> NWLzNXlxRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS0O|I2OzhpPkK0OFczPTN6PD;hQi=>
H460  NWC1Roh[SXCxcITvd4l{KEG|c3H5 NWXmV3VoOTBywrDuUS=> NELySmozPMLiaB?= MnP2bY5lfWOnczDj[YxtKGGyb4D0c5NqeyClbz30doVifGWmIIfpeIghSkWcMkO1 MYe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDR5MkWzPEc,OjR2N{K1N|g9N2F-
GC  NXnk[mZrT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= Mme2OVAuOTJ3IN88US=> M3y1VVczKGh? NGW5doZifHSnboXheIV{KHSqZTDj[YxtKGe{b4f0bEBqdiCjIHTvd4Uu\GWyZX7k[Y51KG2jbn7ldi=> NH7SfFM9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NUe3OFUxOyd-MkW3O|Q2ODN:L3G+
GH3 MVfHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? Mn3lOVAuOTJ3IN88US=> NUX3bmdvPzJiaB?= NY\wVIt1[XS2ZX71ZZRmeyC2aHWgZ4VtdCCpcn;3eIghcW5iYTDkc5NmNWSncHXu[IVvfCCvYX7u[ZI> M4r2NVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ3N{e0OVA{Lz5{NUe3OFUxOzxxYU6=
BT474R  NH3obHJHfW6ldHnvckBCe3OjeR?= NH7uVIc2OCEQvF2= M3\GflExNTZyIHS= NVT6XY56cW6qaXLpeJMhW1SDVEOgZYN1cX[rdIm= MoPVQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjV|Mke1OlEoRjJ3M{K3OVYyRC:jPh?=
NCI-N87R NWi2d4MyTnWwY4Tpc44hSXO|YYm= MkCyOVAh|ryP NGjB[mYyOC14MDDk NIH4SlJqdmirYnn0d{BUXEGWMzDhZ5Rqfmm2eR?= NELMXZU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NUOyO|U3OSd-MkWzNlc2PjF:L3G+
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MDA-MB-231 MoD0SpVv[3Srb36gZZN{[Xl? Mn73NVAxKHWP MUGyOEBpenN? NH;0dWFKdmirYnn0bY9vKG:oIGP0ZZQ{KGGldHn2ZZRqd25iaX6gbJVu[W5iTVTBMW1DNTJ|MTDj[YxteyCjdDCxNFAhfU1iYX\0[ZIhOjRiaILz MmDDQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd2NkOwPVAoRjF5NE[zNFkxRC:jPh?=
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NIH3T3 NFvWVFBHfW6ldHnvckBie3OjeR?= Ml3nNVAxKHWP NWj4bJg6PDhiaILz M{W4PXJm\HWldHnvckBw\iC|dYL2bZZqdiCpZX7lJIV5eHKnc4Ppc44hcW5idj3TdoMhfHKjboPm[YN1\WRibX;1d4UhVkmKM2SzJINmdGy|IHH0JFExOCC3TTDh[pRmeiB2ODDodpM> NIPxO209[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zN{S2N|A6OCd-MUe0OlMxQTB:L3G+
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MDA-MB-231 MlfmSpVv[3Srb36gZZN{[Xl? M{\6U|ExOCC3TR?= MlriOFghcHK| MoTVVoVlfWO2aX;uJI9nKHO3co\peolvKGenbnWg[ZhxemW|c3nvckBqdiCqdX3hckBOTEFvTVKtNlMyKGOnbHzzJIF1KDFyMDD1UUBi\nSncjC0PEBpenN? MnPBQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd2NkOwPVAoRjF5NE[zNFkxRC:jPh?=
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NIH3T3 M4LpV2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M3XxeVExOCC3TR?= NHfIS4VIem:5dHigbY5pcWKrdHnvckBw\iCvb4Xz[UBPUUh|VEOgZ4VtdHNiZYjwdoV{e2mwZzD2MXNz[yCjdDCxNFAhfU1iZn;yJIV3\XK7IEOg[IF6eyCkeTDzc4Z1NWGpYYKgZ49td267LX\vdo1ifGmxbjDhd5NigQ>? NW\wb3VIRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUe0OlMxQTBpPkG3OFY{ODlyPD;hQi=>
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NB1643 MX\xTHRUKGG|c3H5 Ml3TdWhVWyCxZjDw[YRq[XS{aXOgZ4Fv[2W{IHPlcIwhdGmwZYOgeI8hcWSnboTp[pkhdXWudHnwcIUhd3Cyb4L0eY5qfGmnczDmc5Ih\HK3ZzDy[ZB2enCxc3nu[|ohWHKrbXHyfUB{[3KnZX6g[o9zKE6EMU[0N{Bk\Wyucx?= NWThfXRpRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkm0N|UyOzlpPkK5OFM2OTN7PD;hQi=>
LAN-5 M3nM[JFJXFNiYYPzZZk> MlHIdWhVWyCxZjDw[YRq[XS{aXOgZ4Fv[2W{IHPlcIwhdGmwZYOgeI8hcWSnboTp[pkhdXWudHnwcIUhd3Cyb4L0eY5qfGmnczDmc5Ih\HK3ZzDy[ZB2enCxc3nu[|ohWHKrbXHyfUB{[3KnZX6g[o9zKEyDTj21JINmdGy| MnTTQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjl2M{WxN|koRjJ7NEO1NVM6RC:jPh?=
Assay
Methods Test Index PMID
Western blot STAT3 / p-STAT3 / Cyclin D1 / Bcl-2 / Nanog / OCT4 / ALDH1 / CD44 ; PD-L1 26556875 28605599
Immunofluorescence p-STAT3 ; Oct4 / Twist 26556875 27521216
Growth inhibition assay Cell viability 26813676
In vivo S3I-201 (5 mg/kg, i.v. every 2 or every 3 days) shows the antitumor efficacy in mouse models with human breast tumor xenografts that harbor constitutively active Stat3. [1] S3I-201 treatment reduces Varicella-zoster virus (VZV) replication on the basis of the bioluminescence signal and the number of positive skin xenografts compared with DMSO-treated mice by inhibiting STAT3 phosphorylation. [4]

Protocol (from reference)

Kinase Assay:[1]
  • In vitro Stat3 DNA-binding assay and EMSA analysis :

    Briefly, 100 mL of biotinyl-e-Ac-EPQpYEEIEL-OH (in 50 mM Tris/150 mM NaCl, pH 7.5) is added to each well of streptavidin-coated 96-well microtiter plates and incubated with shaking at 4 °C overnight. Then plates are rinsed with PBS/Tween 20 and then two times with 200 mL of BSA-T-PBS (0.2% BSA/0.1% Tween 20/PBS). Then 50 mL of Lck-SH2-GST fusion protein (6.4 ng/ml in BSA-T-PBS) is added to each well of the 96-well plate in the presence and absence of 50 mL of S3I-201 (for 30 and 100 mM final concentrations), and the plate is shaken at room temperature for 4 hours. After solutions are removed, each well is rinsed four times with BSA-T-PBS (200 mL), and 100 mL of polyclonal rabbit anti-GST antibody (100 ng/mL in BSA-T-PBS) is added to each well and incubated at 4 °C overnight. After washing with BSA-T-PBS, 100 mL of 200 ng/mL BSA-T-PBS horseradish peroxidase-conjugated mouse anti-rabbit antibody is added to each well and incubated for 45 minutes at room temperature. After four washing steps with BSA-T-PBS and three washing steps with PBS-T, 100 mL of peroxidase substrate is added to each well and incubated for 5-15 minutes. The peroxidase reaction is stopped by adding 100 mL of 1 M sulfuric acid solution, and absorbance is read at 450 nm with an ELISA plate rea

Cell Research:[2]
  • Cell lines: MDA-MB-435, MDA-MB-453 and MDA-MB-231 cells lines
  • Concentrations: ~ 250 μM
  • Incubation Time: 72 hours
  • Method: The MTT assay is based on the conversion of the yellow tetrazolium salt MTT to purple formazan crystals by metabolically active cells. The MTT assay provides a quantitative determination of viable cells. Cells are seeded in 96-well microplates in complete culture medium in the absence or presence of increasing serial dosages of S3I-201 as indicated. At 72 hours after culture, the number of viable cells is measured by adding 100 μL/well of 2 mg/mL MTT solution. After 2 hours, the medium is removed, and the formazan crystals are dissolved by adding 100 μL dimethylsulfoxide per well. The absorbance is read at 590 nm with an enzyme-linked immunosorbent assay reader. Each treatment point is performed in 10 wells or sextuplicate.
  • (Only for Reference)
Animal Research:[1]
  • Animal Models: Human breast cancer MDA-MB-231 cells are injected s.c. into the left flank of athymic nu/nu mice.
  • Dosages: ≤5 mg/kg
  • Administration: Administered via i.v.
  • (Only for Reference)

Solubility (25°C)

In vitro

DMSO 73 mg/mL
(199.8 mM)
Water Insoluble
Ethanol Insoluble

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
5% DMSO+corn oil
For best results, use promptly after mixing.

3mg/mL

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 365.36
Formula

C16H15NO7S

CAS No. 501919-59-1
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles CC1=CC=C(C=C1)S(=O)(=O)OCC(=O)NC2=CC(=C(C=C2)C(=O)O)O

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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