| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Targets |
Human Endogenous Metabolite
No specific direct pharmacological target; N1,N8-Diacetylspermidine is a metabolite and a disease biomarker. The unlabeled N1,N8-diacetylspermidine is an endogenous polyamine that is produced from spermidine by the action of polyamine N-acetyltransferases (e.g., spermidine N1-acetyltransferase, SSAT). Elevated levels of N1,N8-diacetylspermidine are associated with cancer, but it does not have a direct drug target; instead, it is a metabolic byproduct resulting from increased polyamine biosynthesis and turnover in rapidly proliferating cells (including cancer cells). |
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| ln Vitro |
In vitro, N1,N8-Diacetylspermidine has no direct biological activity of its own; it is a stable end-product of polyamine metabolism. However, its levels are measured as an indicator of cellular proliferation and polyamine metabolism. In cancer cells (e.g., colorectal, bladder, prostate, ovarian, and cervical cancer cell lines), intracellular and extracellular levels of N1,N8-diacetylspermidine are elevated compared to normal cells due to increased activity of ornithine decarboxylase (ODC) and spermidine/spermine N1-acetyltransferase (SSAT). The compound itself is not cytotoxic and does not induce apoptosis or cell cycle arrest; rather, its abundance correlates with the rate of cell proliferation. N1,N8-Diacetylspermidine is stable in urine and plasma and is not further metabolized to a significant extent, making it an ideal biomarker. In in vitro studies using cancer cell lines, treatment with chemotherapeutic agents (e.g., 5-FU, cisplatin, doxorubicin) that induce cell death leads to a transient increase in N1,N8-diacetylspermidine release into the culture medium, reflecting increased polyamine catabolism. Thus, it can be used as a pharmacodynamic biomarker to monitor treatment response in cell culture.
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| ln Vivo |
In vivo, N1,N8-Diacetylspermidine is an endogenous polyamine metabolite found in human urine. Elevated urinary levels of N1,N8-diacetylspermidine serve as a non-invasive biomarker for the early detection, prognosis, and monitoring of treatment response in cancer patients, particularly those with colorectal cancer, bladder cancer, prostate cancer, ovarian cancer, and cervical cancer. In animal models of cancer (e.g., nude mice bearing human colon cancer xenografts), urinary N1,N8-diacetylspermidine levels correlate with tumor burden and decrease following tumor resection or effective chemotherapy. In rats treated with carcinogens (e.g., azoxymethane), elevated urinary N1,N8-diacetylspermidine levels are observed before the appearance of histologically detectable tumors, indicating its potential as an early detection biomarker. The compound itself is not administered for therapeutic purposes; it is measured as an endogenous biomarker. Elevated N1,N8-diacetylspermidine levels are also observed in patients with inflammatory bowel disease (IBD) and other conditions associated with increased cell turnover. In clinical studies, urinary N1,N8-diacetylspermidine has been shown to have high sensitivity (70-90%) and specificity (75-95%) for colorectal cancer detection, often in combination with other polyamines (e.g., N1,N12-diacetylspermine, N1-acetylspermidine) to form a multi-biomarker panel.
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| Enzyme Assay |
No specific enzyme/receptor binding protocol; N1,N8-Diacetylspermidine is analyzed as a biomarker, not as an enzyme/substrate. For quantification of N1,N8-diacetylspermidine in urine, plasma, or tissue homogenates by LC-MS/MS: (1) Prepare a stock solution of N1,N8-Diacetylspermidine (unlabeled) in water or 0.1% formic acid in water at 1 mg/mL. (2) Prepare calibration standards by spiking known concentrations of N1,N8-diacetylspermidine (1-5000 ng/mL) into blank biological matrix (e.g., urine from healthy donors, stripped plasma, or synthetic urine). (3) For sample preparation: mix 50-100 uL of urine or plasma with 150-300 uL of acetonitrile or methanol containing 0.1-1% formic acid. (4) Centrifuge at 14,000 rpm for 10 min at 4degC. (5) Transfer supernatant to a clean tube, evaporate under nitrogen, reconstitute in 50-100 uL of mobile phase (0.1% formic acid in water, or 0.1% formic acid in acetonitrile/water, 5:95). (6) Separate on a HILIC column (e.g., ZIC-HILIC, 2.1 × 100 mm, 3.5 um) or a C18 reverse-phase column using a mobile phase containing heptafluorobutyric acid (HFBA) or pentafluoropropionic acid (PFPA) as ion-pairing reagents to retain polar polyamines. (7) For LC-MS/MS detection: use ESI in positive ion mode. MRM transitions: For N1,N8-Diacetylspermidine: m/z 230 → 171 (or 230 → 100, 230 → 113). (8) For quantification using stable isotope dilution: add a fixed concentration of deuterated internal standard (e.g., N1,N8-Diacetylspermidine-d3 or -d6) to each sample before extraction. (9) Use the analyte/internal standard peak area ratio to calculate concentration via calibration curve. (10) Normalize urinary levels to creatinine concentration (mg/dL) to account for urine dilution. (11) Alternatively, use competitive ELISA kits available for N1,N8-diacetylspermidine, which use a polyclonal antibody specific for the acetylated polyamine.
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| Cell Assay |
(1) For in vitro cell culture experiments: seed cancer cells (e.g., HCT116 colorectal cancer, HT-29, MCF-7, HeLa) in 6-well or 12-well plates at 1 × 10⁶ cells/well in DMEM + 10% FBS. (2) Treat cells with chemotherapeutic agents (e.g., 5-fluorouracil, 5-FU, 10-50 uM; cisplatin, 10-50 uM; doxorubicin, 0.1-1 uM) or experimental compounds for 24-72 h. (3) Collect cell culture supernatant at various time points (24, 48, 72 h). (4) For cellular lysates: wash cells with PBS, add 200 uL of 0.4 M perchloric acid (PCA), scrape cells, centrifuge at 14,000 rpm for 10 min, collect supernatant for polyamine analysis. (5) Analyze N1,N8-Diacetylspermidine levels by LC-MS/MS as described in field 5, or use ELISA. (6) For cell viability: after treatment, add MTT or CCK-8 reagent to the remaining cells, measure viability. Correlate N1,N8-Diacetylspermidine levels in supernatant with cell death/viability. (7) For polyamine metabolism studies: treat cells with DFMO (alpha-difluoromethylornithine, 1-10 mM), an inhibitor of ornithine decarboxylase (ODC), for 48-72 h, then measure N1,N8-Diacetylspermidine levels to assess SSAT activity. (8) For western blot: after treatment, lyse cells, perform SDS-PAGE and blot for ODC, SSAT, and polyamine metabolism-related proteins.
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| Animal Protocol |
(1) For preclinical studies in animal models: use 6-8 week old male athymic nude mice (BALB/c nude, 20-25 g) bearing subcutaneous HCT116 or HT-29 colorectal cancer xenografts (tumor volume 100-200 mm3). (2) Administer chemotherapeutic agents: e.g., 5-fluorouracil (5-FU, 50 mg/kg, IP, weekly) or vehicle (saline) for 14 days. (3) Collect urine samples at baseline (day 0) and at days 3, 7, 10, 14 using metabolic cages (24 h urine collection) or by gentle abdominal massage (spot urine). (4) Collect blood by tail vein or cardiac puncture at endpoint. (5) Measure N1,N8-Diacetylspermidine levels in urine and plasma by LC-MS/MS or ELISA as described. Normalize urinary concentrations to creatinine. (6) Correlate N1,N8-Diacetylspermidine levels with tumor volume (measured by caliper), tumor weight (at necropsy), and treatment response. (7) For early detection studies: use a carcinogen-induced cancer model (e.g., azoxymethane (AOM, 15 mg/kg, IP weekly for 2 weeks) followed by dextran sulfate sodium (DSS, 1-2% in drinking water for 7 days) to induce colorectal cancer in C57BL/6 mice). Collect urine weekly for 10-20 weeks, measure N1,N8-Diacetylspermidine levels, and correlate with histopathological findings (at endpoint: count colon tumors). (8) For human studies: collect first-morning urine samples from cancer patients and healthy controls. (9) Precipitate proteins with acetonitrile (1:1), centrifuge, and analyze by LC-MS/MS or ELISA. (10) Normalize to creatinine: measure creatinine concentration in urine using Jaffe′s method or creatinine assay kit. (11) Perform receiver operating characteristic (ROC) analysis to determine sensitivity, specificity, and cut-off values for cancer detection. (12) For monitoring treatment response: collect urine samples from cancer patients before and after chemotherapy (e.g., neoadjuvant chemotherapy for bladder cancer or colorectal cancer). Compare N1,N8-Diacetylspermidine levels to assess treatment efficacy. Elevated levels generally correlate with tumor burden; successful treatment leads to decreased levels.
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| ADME/Pharmacokinetics |
Standard formulation for N1,N8-Diacetylspermidine as an analytical standard: prepare a stock solution in water or 0.1% formic acid in water at 1 mg/mL. For LC-MS analysis, dilute stock in 0.1% formic acid in water to prepare calibration standards (1-5000 ng/mL). For ELISA, follow kit instructions. Storage: store powder at -20degC for 3 years, protect from light. In solution (water, 0.1% formic acid), store at -20degC for up to 6 months, -80degC for up to 1 year; avoid repeated freeze-thaw cycles. Solubility: N1,N8-Diacetylspermidine (free base) is a small polar molecule (M.W. 229.32); it is soluble in water (10-50 mg/mL) and in organic solvents such as methanol, ethanol, and DMSO (≥ 20 mg/mL). The compound is stable in urine for at least 6 months at -80degC; at 4degC, stable for 7 days; at room temperature, stable for 24 h. For long-term storage of urine samples, acidify with 0.1 M HCl to pH 3-4 to prevent bacterial degradation, then store at -80degC. Endogenous levels in humans: Urinary N1,N8-Diacetylspermidine levels in healthy adults: typically 0.1-1.0 umol/g creatinine; in colorectal cancer patients, levels may be elevated to 1-10 umol/g creatinine. In plasma, levels are lower (nM range).
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| Toxicity/Toxicokinetics |
In vitro toxicity: N1,N8-Diacetylspermidine is an endogenous polyamine metabolite and is non-toxic at physiological concentrations (uM to low mM). In vitro, up to 1 mM concentrations do not cause cytotoxicity in HEK293 or cancer cells (cell viability >90% by MTT for 48 h). In vivo toxicity: not applicable, as it is an endogenous metabolite present in urine and tissues at low concentrations (uM range). The compound is not administered for therapeutic effect. The synthetic standard used for research is non-toxic at the concentrations used for analytical purposes (ng/mL to ug/mL). The compound is for research use only, not for human therapeutic use. Standard laboratory safety precautions: gloves, lab coat, eye protection. No teratogenic, mutagenic, or reproductive toxicity data is available, but based on its endogenous nature, it is unlikely to be toxic.
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| References |
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| Additional Infomation |
N-{3-[(4-acetaminobutyl)amino]propyl}acetamide is a member of the acetamide class of compounds.
N1,N8-Diacetylspermidine is an endogenous polyamine metabolite found in human urine, formed by the acetylation of spermidine by spermidine/spermine N1-acetyltransferase (SSAT). It serves as a non-invasive biomarker for the early detection, prognosis, and monitoring of treatment response in various cancers, including colorectal cancer, bladder cancer, prostate cancer, ovarian cancer, and cervical cancer. Elevated urinary levels of N1,N8-diacetylspermidine are associated with increased polyamine turnover in rapidly proliferating tumor cells. This compound is not a drug and has no direct pharmacological activity; it is used purely as an analytical standard and a research biomarker. It is not FDA-approved for clinical diagnosis but is used as a research tool in cancer biology, clinical chemistry, and metabolomics. The compound is intended for laboratory research use only and is not for therapeutic or diagnostic applications without appropriate regulatory authorization. For research use, the synthetic standard (≥98% purity) is used to quantify endogenous levels by LC-MS/MS, GC-MS, or ELISA. The diacetylated form is more stable than spermidine and is resistant to further metabolism, making it an ideal biomarker for polyamine metabolism. This product is for research purposes only. |
| Molecular Formula |
C11H23N3O2
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|---|---|
| Molecular Weight |
229.32
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| Exact Mass |
229.179
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| CAS # |
82414-35-5
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| Related CAS # |
N1,N8-Diacetylspermidine hydrochloride;178244-42-3
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| PubChem CID |
389613
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| Appearance |
White to off-white solid powder
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| LogP |
1.191
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
16
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| Complexity |
207
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(NCCCCNCCCNC(=O)C)(=O)C
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| InChi Key |
BKCVMAZDKFQPHB-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H23N3O2/c1-10(15)13-8-4-3-6-12-7-5-9-14-11(2)16/h12H,3-9H2,1-2H3,(H,13,15)(H,14,16)
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| Chemical Name |
N-[4-(3-acetamidopropylamino)butyl]acetamide
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.3607 mL | 21.8036 mL | 43.6072 mL | |
| 5 mM | 0.8721 mL | 4.3607 mL | 8.7214 mL | |
| 10 mM | 0.4361 mL | 2.1804 mL | 4.3607 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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