WARNING: This product is for research use only, not for human or veterinary use.
MedKoo CAT#: 406308
CAS#: 675576-98-4 (4S5R)
Description: Nutlin-3a, also known as SML 0580, is an inhibitor of MDM2 (human homolog of murine double minute 2), which disrupts its interaction with p53, leading to the stabilization and activation of p53. Nutlin-3a activates the p53 pathway and efficiently induces apoptosis in tumours with amplified MDM2 gene and overexpression of MDM2 protein. Nutlin-3 enhances the bortezomib sensitivity of p53-defective cancer cells by inducing paraptosis.
MedKoo Cat#: 406308
Name: Nutlin-3a
CAS#: 675576-98-4 (4S5R)
Chemical Formula: C30H30Cl2N4O4
Exact Mass: 580.16441
Molecular Weight: 581.49
Elemental Analysis: C, 61.97; H, 5.20; Cl, 12.19; N, 9.64; O, 11.01
Related CAS #: 675576-98-4 (4S5R) 548472-68-0 (4R5S and 4S5R) 675576-97-3 (4R5S)
Synonym: Nutlin3a; Nutlin 3a; Nutlin-3a; SML 0580; SML0580; SML-0580; (-)-Nutlin 3, (-)-Nutlin-3;
IUPAC/Chemical Name: 4-((4S,5R)-4,5-bis(4-chlorophenyl)-2-(2-isopropoxy-4-methoxyphenyl)-4,5-dihydro-1H-imidazole-1-carbonyl)piperazin-2-one
InChi Key: BDUHCSBCVGXTJM-WUFINQPMSA-N
InChi Code: InChI=1S/C30H30Cl2N4O4/c1-18(2)40-25-16-23(39-3)12-13-24(25)29-34-27(19-4-8-21(31)9-5-19)28(20-6-10-22(32)11-7-20)36(29)30(38)35-15-14-33-26(37)17-35/h4-13,16,18,27-28H,14-15,17H2,1-3H3,(H,33,37)/t27-,28+/m0/s1
SMILES Code: O=C1NCCN(C(N2[C@H](C3=CC=C(Cl)C=C3)[C@H](C4=CC=C(Cl)C=C4)N=C2C5=CC=C(OC)C=C5OC(C)C)=O)C1
Appearance: Solid powder
Purity: >98% (or refer to the Certificate of Analysis)
Shipping Condition: Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition: Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility: Soluble in DMSO, not in water
Shelf Life: >2 years if stored properly
Drug Formulation: This drug may be formulated in DMSO
Stock Solution Storage: 0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code: 2934.99.9001
| Biological target: | Nutlin-3a, an active enantiomer of Nutlin-3, is a potent murine double minute (MDM2) inhibitor (IC50=90 nM). |
| In vitro activity: | Fifteen epithelial ovarian cancer cell lines of varying histologic subtypes were treated with Nutlin-3a with determination of IC50 values. Western Blot (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) analyses quantified MDM2, p53, and p21 expression after Nutlin-3a treatment. DNA from 15 cell lines was then sequenced for TP53 mutations in exons 2-11 including intron-exon boundaries. Responses to Nutlin-3a were dependent upon TP53 mutation status. By qRT-PCR and WB, levels of MDM2 and p21 were upregulated in wild-type TP53 sensitive cell lines, and p21 induction was reduced or absent in mutant cell lines. Annexin V assays demonstrated apoptosis in sensitive cell lines treated with Nutlin-3a. Reference: PLoS One. 2015 Aug 6;10(8):e0135101. https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/26248031/ |
| In vivo activity: | To increase its potency the active enantiomer nutlin-3a was purified and a 3-week antitumor efficacy study in SJSA-1 tumor-bearing nude mice was performed(Fig. 6A). Nutlin-3a suppressed xenograft growth in a dose-dependent fashion with the highest dose (200 mg/kg) showing a substantial tumor shrinkage (eight partial and one full regressions). No weight loss or significant pathological changes were recorded during the course of the study (data not shown). To show that the antitumor effect of nutlin-3a is caused by activation of the p53 pathway we analyzed the level of p53 targets p21 and MDM2 in nutlin-treated SJSA-1 xenografts. Western blot analysis revealed accumulation of both proteins in tumors from nutlin-treated animals but not vehicle controls (Fig. 6B). This experiment is consistent with the hypothesis that the in vivo antitumor effect of nutlin-3 is derived from activation of the p53 pathway. To exclude any possibility for an in vivo-related artifact we also tested nutlin-3a on cells with mutant p53. Colon cancer cell line HT29 expresses mutant p53 and does not respond to nutlin-3a treatment in vitro (data not shown). Nude mice bearing established HT29 xenografts were treated orally with 200 mg/kg nutlin-3 for 3 weeks. In agreement with the in vitro results, HT29 xenografts showed undistinguishable growth characteristics in the presence or absence of nutlin-3 (data not shown). These experiments confirmed that nutlin-3 is a selective activator of the p53 pathway in vivo and highly efficacious against SJSA-1 osteosarcoma tumors. Reference: Proc Natl Acad Sci U S A. 2006 Feb 7;103(6):1888-93. https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/16443686/ |
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMSO | 25.0 | 43.0 |
The following data is based on the product molecular weight 581.49 Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass | 1 mg | 5 mg | 10 mg |
|---|---|---|---|
| 1 mM | 1.15 mL | 5.76 mL | 11.51 mL |
| 5 mM | 0.23 mL | 1.15 mL | 2.3 mL |
| 10 mM | 0.12 mL | 0.58 mL | 1.15 mL |
| 50 mM | 0.02 mL | 0.12 mL | 0.23 mL |
| In vitro protocol: | 1. Crane EK, Kwan SY, Izaguirre DI, Tsang YT, Mullany LK, Zu Z, Richards JS, Gershenson DM, Wong KK. Nutlin-3a: A Potential Therapeutic Opportunity for TP53 Wild-Type Ovarian Carcinomas. PLoS One. 2015 Aug 6;10(8):e0135101. doi: 10.1371/journal.pone.0135101. PMID: 26248031; PMCID: PMC4527847. 2. Kobayashi M, Ishizaki Y, Owaki M, Matsumoto Y, Kakiyama Y, Hoshino S, Tagawa R, Sudo Y, Okita N, Akimoto K, Higami Y. Nutlin-3a suppresses poly (ADP-ribose) polymerase 1 by mechanisms different from conventional PARP1 suppressors in a human breast cancer cell line. Oncotarget. 2020 May 5;11(18):1653-1665. doi: 10.18632/oncotarget.27581. PMID: 32405340; PMCID: PMC7210013. |
| In vivo protocol: | 1. Tovar C, Rosinski J, Filipovic Z, Higgins B, Kolinsky K, Hilton H, Zhao X, Vu BT, Qing W, Packman K, Myklebost O, Heimbrook DC, Vassilev LT. Small-molecule MDM2 antagonists reveal aberrant p53 signaling in cancer: implications for therapy. Proc Natl Acad Sci U S A. 2006 Feb 7;103(6):1888-93. doi: 10.1073/pnas.0507493103. Epub 2006 Jan 27. PMID: 16443686; PMCID: PMC1413632. 2. Lerche CM, Philipsen PA, Poulsen T, Gniadecki R, Wulf HC. Topical nutlin-3a does not decrease photocarcinogenesis induced by simulated solar radiation in hairless mice. Photodermatol Photoimmunol Photomed. 2012 Aug;28(4):207-12. doi: 10.1111/j.1600-0781.2012.00675.x. PMID: 23017174. |
1: Yee-Lin V, Pooi-Fong W, Soo-Beng AK. Nutlin-3, A p53-Mdm2 Antagonist for Nasopharyngeal Carcinoma Treatment. Mini Rev Med Chem. 2018;18(2):173-183. doi: 10.2174/1389557517666170717125821. PMID: 28714398; PMCID: PMC5769085.
2: Wang X, Chen Y, Liu H, Guo S, Hu Y, Zhan Y, An H. A novel anti-cancer mechanism of Nutlin-3 through downregulation of Eag1 channel and PI3K/AKT pathway. Biochem Biophys Res Commun. 2019 Sep 24;517(3):445-451. doi: 10.1016/j.bbrc.2019.07.106. Epub 2019 Aug 1. PMID: 31378368.
3: Wang M, Wang X, Li Y, Xiao Q, Cui XH, Xiao GD, Wang JC, Xu CW, Ren H, Liu D. Nutlin-3-Induced Sensitization of Non-Small Cell Lung Cancer Stem Cells to Axitinib-Induced Apoptosis Through Repression of Akt1/Wnt Signaling. Oncol Res. 2019 Sep 23;27(9):987-995. doi: 10.3727/096504018X15424918479652. Epub 2019 Mar 4. PMID: 30832755; PMCID: PMC7848271.
4: He T, Guo J, Song H, Zhu H, Di X, Min H, Wang Y, Chen G, Dai W, Ma J, Sun X, Ma J. Nutlin-3, an Antagonist of MDM2, Enhances the Radiosensitivity of Esophageal Squamous Cancer with Wild-Type p53. Pathol Oncol Res. 2018 Jan;24(1):75-81. doi: 10.1007/s12253-017-0215-5. Epub 2017 Mar 24. PMID: 28341911.
5: Lee DM, Kim IY, Seo MJ, Kwon MR, Choi KS. Nutlin-3 enhances the bortezomib sensitivity of p53-defective cancer cells by inducing paraptosis. Exp Mol Med. 2017 Aug 11;49(8):e365. doi: 10.1038/emm.2017.112. PMID: 28798402; PMCID: PMC5579507.
6: Wu L , Hu Y , He Y , Xia Y , Lu H , Cao Z , Yi X , Wang J . Dual-channel surface plasmon resonance monitoring of intracellular levels of the p53-MDM2 complex and caspase-3 induced by MDM2 antagonist Nutlin-3. Analyst. 2019 Jul 7;144(13):3959-3966. doi: 10.1039/c9an00301k. Epub 2019 May 28. PMID: 31134974.
7: Drost J, van Jaarsveld RH, Ponsioen B, Zimberlin C, van Boxtel R, Buijs A, Sachs N, Overmeer RM, Offerhaus GJ, Begthel H, Korving J, van de Wetering M, Schwank G, Logtenberg M, Cuppen E, Snippert HJ, Medema JP, Kops GJ, Clevers H. Sequential cancer mutations in cultured human intestinal stem cells. Nature. 2015 May 7;521(7550):43-7. doi: 10.1038/nature14415. Epub 2015 Apr 29. PMID: 25924068.
8: Secchiero P, Bosco R, Celeghini C, Zauli G. Recent advances in the therapeutic perspectives of Nutlin-3. Curr Pharm Des. 2011;17(6):569-77. doi: 10.2174/138161211795222586. PMID: 21391907.
9: Voltan R, Secchiero P, Ruozi B, Caruso L, Forni F, Palomba M, Zauli G, Vandelli MA. Nanoparticles loaded with Nutlin-3 display cytotoxicity towards p53(wild-type) JVM-2 but not towards p53(mutated) BJAB leukemic cells. Curr Med Chem. 2013;20(21):2712-22. doi: 10.2174/0929867311320210007. PMID: 23627936.
10: Wu Q, Wang X, Liu J, Zheng J, Liu Y, Li Y, Su F, Ou W, Wang R. Nutlin-3 reverses the epithelial-mesenchymal transition in gemcitabine-resistant hepatocellular carcinoma cells. Oncol Rep. 2016 Sep;36(3):1325-32. doi: 10.3892/or.2016.4920. Epub 2016 Jul 8. PMID: 27430152.
11: Bajelan B, Zaki-Dizaji M, Darabi S, Rajaei F. The effects of Nutlin-3 on morphology, cellular proliferation, and apoptosis in rat primary mesenchymal stem cells. J Cell Physiol. 2019 Jul;234(7):11424-11430. doi: 10.1002/jcp.27798. Epub 2018 Nov 27. PMID: 30478967.
12: Wang XH, Zhang SF, Bao JT, Liu FY. Oridonin synergizes with Nutlin-3 in osteosarcoma cells by modulating the levels of multiple Bcl-2 family proteins. Tumour Biol. 2017 Jun;39(6):1010428317701638. doi: 10.1177/1010428317701638. PMID: 28618955.
13: Bajelan B, Zaki-Dizaji M, Rahmani B, Darzi S, Darabi S, Rajaei F. Resistance of human primary mesenchymal stem cells to cytotoxic effects of nutlin-3 in vitro. J Cell Biochem. 2020 Jan;121(1):788-796. doi: 10.1002/jcb.29324. Epub 2019 Aug 26. PMID: 31452266.
14: Miao R, Xu X, Wang Z, Liu S, Qu K, Chen W, Liu C. Synergistic effect of nutlin-3 combined with aspirin in hepatocellular carcinoma HepG2 cells through activation of Bcl-2/Bax signaling pathway. Mol Med Rep. 2018 Mar;17(3):3735-3743. doi: 10.3892/mmr.2017.8346. Epub 2017 Dec 22. PMID: 29286113; PMCID: PMC5802178.
15: Lee SY, Choi HC, Choe YJ, Shin SJ, Lee SH, Kim HS. Nutlin-3 induces BCL2A1 expression by activating ELK1 through the mitochondrial p53-ROS-ERK1/2 pathway. Int J Oncol. 2014 Aug;45(2):675-82. doi: 10.3892/ijo.2014.2463. Epub 2014 May 26. PMID: 24867259.
16: Voon YL, Ahmad M, Wong PF, Husaini R, Ng WT, Leong CO, Lane DP, Khoo AS. Nutlin-3 sensitizes nasopharyngeal carcinoma cells to cisplatin-induced cytotoxicity. Oncol Rep. 2015 Oct;34(4):1692-700. doi: 10.3892/or.2015.4177. Epub 2015 Aug 5. PMID: 26252575; PMCID: PMC4564086.
17: Guo Y, Li Y, Xiang B, Huang XO, Ma HB, Wang FF, Gong YP. Nutlin-3 plus tanshinone IIA exhibits synergetic anti-leukemia effect with imatinib by reactivating p53 and inhibiting the AKT/mTOR pathway in Ph+ ALL. Biochem J. 2017 Dec 6;474(24):4153-4170. doi: 10.1042/BCJ20170386. PMID: 29046392.
18: Chen H, Xue L, Huang H, Wang H, Zhang X, Zhu W, Wang Z, Wang Z, Wu H. Synergistic effect of Nutlin-3 combined with MG-132 on schwannoma cells through restoration of merlin and p53 tumour suppressors. EBioMedicine. 2018 Oct;36:252-265. doi: 10.1016/j.ebiom.2018.09.042. Epub 2018 Sep 28. PMID: 30274821; PMCID: PMC6197711.
19: Choe YJ, Lee SY, Ko KW, Shin SJ, Kim HS. Nutlin-3 induces HO-1 expression by activating JNK in a transcription-independent manner of p53. Int J Oncol. 2014 Mar;44(3):761-8. doi: 10.3892/ijo.2013.2227. Epub 2013 Dec 23. PMID: 24366007.
20: Vatsyayan R, Singhal J, Nagaprashantha LD, Awasthi S, Singhal SS. Nutlin-3 enhances sorafenib efficacy in renal cell carcinoma. Mol Carcinog. 2013 Jan;52(1):39-48. doi: 10.1002/mc.20875. Epub 2011 Oct 17. PMID: 22006587; PMCID: PMC4153352.
Nutlins are cis-imidazoline analogs which inhibit the interaction between MDM2 and tumour suppressor p53, and were discovered by screening a chemical library by Vassilev et al. Nutlin-1, Nutlin-2 and Nutlin-3 were all identified in the same screen, however Nutlin-3 is the compound most commonly used in anti-cancer studies. Inhibiting the interaction between mdm2 and p53 stabilizes p53 and is thought to selectively induce a growth-inhibiting state called senescence in cancer cells. These compounds are therefore thought to work best on tumors that contain normal or wild type p53.[citation needed] Nutlin-3 has been shown to affect the production of p53 within minutes. (source: http://en.wikipedia.org/wiki/Nutlin).
