| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
Phosphoramide mustard cyclohexanamine targets DNA, specifically alkylating guanine residues. The compound induces DNA crosslinking and damage, forming cross-linked DNA adducts that hinder DNA strand separation during replication. It also increases the production of covalent DNA-protein conjugates. These DNA lesions lead to impaired DNA replication, cell cycle arrest, and cell death, particularly in rapidly dividing cancer cells.
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| ln Vitro |
In order to prevent DNA strand separation during replication, phosphoramide mustard cyclohexanamine forms cross-linked DNA adducts, which are cytotoxic[1]. For a 48-hour period, rat spontaneously immortalized granulosa cells (SIGCs) exposed to phosphoramide mustard cyclohexanamine (3-6 μM) lose viability. Damage to ovarian DNA and the production of DNA adducts are caused by phosphoamide mustard cyclohexanamine (3-6 μM; 24-48 hours). The expression of DNA damage response (DDR) gene mRNA and DDR proteins are increased by phosphoamide mustard cyclohexanamine (3-6 μM; 24-48 hours)[1].
In vitro, Phosphoramide mustard cyclohexanamine induces DNA cross-linking and damage in SIGC (rat ovarian granulosa) cells, reducing cell viability in a dose-dependent manner. Cell viability is significantly decreased at concentrations of 3 microM and 6 microM after 48 hours of exposure. The compound is cytotoxic to HT-1080 human fibrosarcoma cells in a concentration-dependent manner. It reduces cell viability and increases DNA damage response (DDR) gene expression and protein levels in SIGCs. |
| ln Vivo |
Rats' subcutaneous tumor growth is inhibited by phosphoramide mustard cyclohexanamine (2.1-20.7 mg/kg; ip; daily; for 5 days)[2]. After intravenous injection (rat 59.4 mg/kg), phosphoramide mustard cyclohexanamine shows terminal elimination half-lives (rat 15.1 min)[2].
In vivo, Phosphoramide mustard cyclohexanamine (2.1-20.7 mg/kg; intraperitoneal; daily for 5 days) inhibits subcutaneous tumor growth in rats. The compound exhibits significant anticancer activity by inhibiting tumor growth in animal models. It is a major contributor to cyclophosphamide's anticancer activity. The compound activates the ovarian DNA repair response in granulosa cells. |
| Enzyme Assay |
In vitro DNA binding assays for Phosphoramide mustard cyclohexanamine involve incubating the compound with purified DNA or oligonucleotides. DNA alkylation and crosslink formation are analyzed by gel electrophoresis, denaturation, or HPLC-MS. Guanine alkylation at the N7 position is confirmed by mass spectrometry. DNA-protein conjugate formation is assessed by immunoblotting or pull-down assays. Dose-response experiments determine the concentration-dependent effects on DNA damage.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: SIGCs Tested Concentrations: 0.5 μM, 1 μM, 3 μM, 6 μM Incubation Duration: 48 hrs (hours) Experimental Results: decreased cell viability at concentrations of 3 μM and higher. RT-PCR[1] Cell Types: SIGCs Tested Concentrations: 3 μM, 6 μM Incubation Duration: 24 hrs (hours), 48 hrs (hours) Experimental Results: Increased DDR gene mRNA expression levels. Western Blot Analysis[1] Cell Types: SIGCs Tested Concentrations: 3 μM, 6 μM Incubation Duration: 24 hrs (hours), 48 hrs (hours) Experimental Results: Generally increased DDR proteins. In vitro cell-based assays for Phosphoramide mustard cyclohexanamine use SIGC (rat ovarian granulosa) cells, HT-1080 human fibrosarcoma cells, or other cancer cell lines. Cells are treated with compound at concentrations ranging from 0-10 microM for 24-48 hours. Cell viability is assessed by MTT or CCK-8 assays. DNA damage response is evaluated by Western blot for phosphorylated H2AX (gamma-H2AX), ATM, and other DDR markers. Gene expression changes are analyzed by qPCR. |
| Animal Protocol |
Animal/Disease Models: Rat, subcutaneously (sc) implanted Walker 256 carcinosarcoma tumor[2]
Doses: 2.1 mg/kg, 4.8 mg/kg , 10.4 mg/kg, 20.7 mg/kg Route of Administration: intraperitoneal (ip)injection, one time/day, for 5 days Experimental Results: Required to produce 50% inhibition of subcutaneous (sc)tumor growth with dose of 12 mg/kg. Animal/Disease Models: Rats[2] Doses: 86.0 mg/kg (pharmacokinetic/PK Analysis) Route of Administration: intravenous (iv)injection Experimental Results: T1/2 (15.1 min). In vivo animal studies for Phosphoramide mustard cyclohexanamine typically use rat or mouse tumor models. The compound is administered intraperitoneally at doses of 2.1-20.7 mg/kg daily for 5 days. Subcutaneous tumor growth is monitored by caliper measurements. Pharmacokinetic studies involve intravenous administration (86.0 mg/kg) with plasma sampling to determine half-life and clearance. Tissue distribution and DNA adduct formation may be assessed post-mortem. |
| ADME/Pharmacokinetics |
Pharmacokinetic properties: Phosphoramide mustard cyclohexanamine (86.0 mg/kg; i.v.) has a plasma disappearance half-life of 15.1 minutes in rats. Plasma concentration decline is biphasic, with a longer terminal half-life of 8.68 hours. Renal function influences total exposure. The compound is the final cytotoxic metabolite formed from cyclophosphamide through hepatic metabolism. Plasma concentrations following therapeutic cyclophosphamide doses are probably sufficient for cytotoxic effects.
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| Toxicity/Toxicokinetics |
Toxicity of Phosphoramide mustard cyclohexanamine is characteristic of alkylating agents. It induces DNA damage, which can lead to cytotoxicity in normal rapidly dividing cells such as bone marrow and gastrointestinal epithelium. Myelosuppression is a expected toxicity. The compound is a genotoxic agent and potential carcinogen. It is for research use only and not for human therapeutic use. Standard safety precautions for handling alkylating agents apply, including use of fume hoods and personal protective equipment.
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| References |
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| Additional Infomation |
Phosphoramide mustard cyclohexanamine is the major active metabolite of cyclophosphamide. It exerts its cytotoxic effects through DNA crosslinking, guanine alkylation, and formation of DNA-protein conjugates. The compound destroys rapidly dividing cells and leads to DNA damage. The cyclohexanamine moiety may modify pharmacokinetic or lipophilic properties, potentially enhancing tissue penetration or altering toxicity. It is used in the development of chemotherapeutic drugs for cancer treatment.
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| Molecular Formula |
C10H24CL2N3O2P
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|---|---|
| Molecular Weight |
320.20
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| Exact Mass |
319.098
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| Elemental Analysis |
C, 37.51; H, 7.56; Cl, 22.14; N, 13.12; O, 9.99; P, 9.67
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| CAS # |
1566-15-0
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| Related CAS # |
Phosphoramide mustard;10159-53-2
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| PubChem CID |
96355
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| Appearance |
White to off-white solid powder
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| Boiling Point |
363.5ºC at 760mmHg
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| Melting Point |
100-103 °C
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| Flash Point |
173.6ºC
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| Vapour Pressure |
2.84E-06mmHg at 25°C
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| LogP |
3.503
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
18
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| Complexity |
197
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1(N)CCCCC1.ClCCN(CCCl)P(N)(=O)O
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| InChi Key |
BGTIPRUDEMNRIP-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C6H13N.C4H11Cl2N2O2P/c7-6-4-2-1-3-5-6;5-1-3-8(4-2-6)11(7,9)10/h6H,1-5,7H2;1-4H2,(H3,7,9,10)
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| Chemical Name |
amino-[bis(2-chloroethyl)amino]phosphinic acid;cyclohexanamine
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| Synonyms |
PMC; NSC-69945; NSC 69945; NSC69945; Phosphoramide Mustard cyclohexylammonium salt
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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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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) |
H2O : ~100 mg/mL (~312.3 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (312.30 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1230 mL | 15.6152 mL | 31.2305 mL | |
| 5 mM | 0.6246 mL | 3.1230 mL | 6.2461 mL | |
| 10 mM | 0.3123 mL | 1.5615 mL | 3.1230 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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