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Physicochemical Properties
| Molecular Formula | C69H122N26O22 |
| Molecular Weight | 1667.86798 |
| Exact Mass | 1666.92 |
| CAS # | 144796-71-4 |
| Related CAS # | TNF-α (31-45), human TFA |
| PubChem CID | 71311650 |
| Appearance | White to off-white solid powder |
| LogP | 1.697 |
| Hydrogen Bond Donor Count | 26 |
| Hydrogen Bond Acceptor Count | 26 |
| Rotatable Bond Count | 57 |
| Heavy Atom Count | 117 |
| Complexity | 3550 |
| Defined Atom Stereocenter Count | 14 |
| SMILES | C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)N |
| InChi Key | HYFATHUJEOQTRM-PJKQPBPGSA-N |
| InChi Code | InChI=1S/C69H122N26O22/c1-30(2)23-41(93-64(114)42(24-31(3)4)89-53(103)35(10)85-62(112)45(27-48(72)97)91-55(105)34(9)83-58(108)38(16-13-21-80-68(75)76)86-56(106)37(70)15-12-20-79-67(73)74)61(111)84-36(11)54(104)90-44(26-47(71)96)57(107)82-29-49(98)95-52(33(7)8)65(115)88-40(18-19-50(99)100)60(110)92-43(25-32(5)6)63(113)87-39(17-14-22-81-69(77)78)59(109)94-46(66(116)117)28-51(101)102/h30-46,52H,12-29,70H2,1-11H3,(H2,71,96)(H2,72,97)(H,82,107)(H,83,108)(H,84,111)(H,85,112)(H,86,106)(H,87,113)(H,88,115)(H,89,103)(H,90,104)(H,91,105)(H,92,110)(H,93,114)(H,94,109)(H,95,98)(H,99,100)(H,101,102)(H,116,117)(H4,73,74,79)(H4,75,76,80)(H4,77,78,81)/t34-,35-,36-,37-,38-,39-,40-,41-,42-,43-,44-,45-,46-,52-/m0/s1 |
| Chemical Name | (2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-5-(diaminomethylideneamino)pentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]propanoyl]amino]-4-oxobutanoyl]amino]propanoyl]amino]-4-methylpentanoyl]amino]-4-methylpentanoyl]amino]propanoyl]amino]-4-oxobutanoyl]amino]acetyl]amino]-3-methylbutanoyl]amino]-4-carboxybutanoyl]amino]-4-methylpentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]butanedioic acid |
| 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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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
| ln Vitro | Treatment of adenoid cystic carcinoma (ACC) cell lines (ACC-83 and ACC-LM) with 5 µM TNF-α (31-45), human for 1 hour activated the NF-κB pathway, as evidenced by increased phosphorylation of the P65 subunit. This activation promoted cisplatin resistance and enhanced the migration and invasion capabilities of the ACC cells. The use of TNF-α (31-45), human did not alter the mRNA or protein expression levels of RPS3. [2] |
| Cell Assay |
NF-κB Pathway Activation Assay: To activate the NF-κB pathway, ACC cells were treated with TNF-α (31-45), human at a final concentration of 5 µM for 1 hour. The activation status of the pathway was subsequently assessed by western blot analysis of phosphorylated P65 (P-P65) levels. [2] Cisplatin Resistance (IC₅₀) Assay: Cells were seeded into 96-well plates at 10,000 cells/well. After treatment with TNF-α (31-45), human (5 µM, 1 hour) or other conditions, cisplatin was added at a series of concentrations (0, 2.5, 5, 10, 20, 40, 80, 160 µM) for 48 hours. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay to determine the half-maximal inhibitory concentration (IC₅₀) of cisplatin. [2] Migration and Invasion Assay (Transwell): For migration assays, 30,000 cells in medium containing 1% FBS were seeded into the upper chamber of a Transwell insert (8-µm pore size). For invasion assays, the upper chamber was coated with matrix gel. The lower chamber contained complete medium with 10% FBS. After pretreatment with TNF-α (31-45), human (5 µM, 1 hour), cells were allowed to migrate or invade for 24 hours. Cells on the lower surface of the membrane were fixed, stained with crystal violet, and counted under a microscope. [2] |
| References |
[1]. Idriss HT, Naismith JH. TNF alpha and the TNF receptor superfamily: structure-function relationship(s). Microsc Res Tech. 2000 Aug 1;50(3):184-95. [2]. RPS3 Promotes the Metastasis and Cisplatin Resistance of Adenoid Cystic Carcinoma. Front Oncol. 2022 Jun 30:12:804439. [3]. The Role of Tumor Necrosis Factor Alpha (TNF-α) in Autoimmune Disease and Current TNF-α Inhibitors in Therapeutics. Int J Mol Sci. 2021 Mar 8;22(5):2719. |
| Additional Infomation |
TNF-α Biology: Tumor necrosis factor alpha (TNF-α) is a homotrimeric cytokine with pleiotropic effects, primarily known as a key regulator of inflammatory responses. It exists in a soluble form (sTNF-α) and a transmembrane precursor form (tmTNF-α). Its biological activities are mediated through two receptors: TNFR1 (expressed ubiquitously, contains a death domain, primarily pro-inflammatory and pro-apoptotic) and TNFR2 (expressed mainly on immune cells, promotes cell survival, proliferation, and tissue regeneration). Dysregulated or excessive TNF-α signaling is implicated in the pathogenesis of various autoimmune diseases. [1] Role in Autoimmune Diseases: The review details the pathogenic role of excessive TNF-α in several autoimmune diseases: In Rheumatoid Arthritis (RA), it activates synovial fibroblasts and osteoclasts, leading to joint destruction. In Psoriatic Arthritis (PsA) and Psoriasis (PS), it drives keratinocyte proliferation and inflammatory cell recruitment. In Inflammatory Bowel Disease (IBD), it contributes to intestinal fibrosis, matrix degradation, and epithelial damage. In Noninfectious Uveitis (NIU), it promotes T cell differentiation and leukocyte infiltration into the eye. [1] Therapeutic TNF-α Inhibitors: The article reviews several clinically approved biologic drugs that inhibit TNF-α, but none are the peptide “TNF-α (31-45), human”. These include: Infliximab (chimeric monoclonal antibody), Etanercept (TNFR2-Fc fusion protein), Adalimumab (fully human monoclonal antibody), Certolizumab Pegol (PEGylated Fab’ fragment), and Golimumab (fully human monoclonal antibody). Their mechanisms involve blocking the interaction of TNF-α with its receptors (TNFR1/2). The review also lists FDA-approved biosimilars for some of these originator drugs and mentions new anti-TNF agents (e.g., Ozoralizumab) under development. [1] Important Note: The molecule “TNF-α (31-45), human” is not discussed, studied, or mentioned in this review article. The article focuses on the endogenous TNF-α cytokine as a pathogenic factor and on protein-based therapeutic agents that block its activity. [1] In this study, TNF-α (31-45), human is employed solely as a pharmacological tool to activate the NF-κB signaling pathway in vitro, in order to investigate the role of this pathway in regulating cisplatin resistance and metastatic behavior in adenoid cystic carcinoma cells. [2] |
Solubility Data
| Solubility (In Vitro) | H2O : ~50 mg/mL (~29.98 mM) |
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.5996 mL | 2.9978 mL | 5.9957 mL | |
| 5 mM | 0.1199 mL | 0.5996 mL | 1.1991 mL | |
| 10 mM | 0.0600 mL | 0.2998 mL | 0.5996 mL |