| Size | Price | Stock | Qty |
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| 50mg |
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| Other Sizes |
| Targets |
XL388-C2-amide-PEG9-NH2 hydrochloride itself is not an active drug compound but a chemical intermediate. The intended target of the final synthesized product (C26-linked rapamycin analog) is the mechanistic target of rapamycin (mTOR), a serine/threonine protein kinase that regulates cell growth, proliferation, metabolism, and autophagy. Rapamycin and its analogs (rapalogs) bind to the FK506-binding protein 12 (FKBP12), and the resulting complex binds to and inhibits the mTOR complex 1 (mTORC1). The C26 position of rapamycin is a site that can be chemically modified without losing binding to FKBP12, allowing for the synthesis of novel rapamycin analogs with altered pharmacokinetic or targeting properties. The PEG9 linker in this intermediate provides increased solubility and flexibility, while the terminal NH2 group allows for conjugation to various reporter groups (e.g., biotin, fluorescent dyes, peptides, or antibodies). Thus, the final products derived from this intermediate target mTOR, a central regulator of cell growth and metabolism.
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| ln Vitro |
The intermediate XL388-C2-amide-PEG9-NH2 hydrochloride itself is not biologically active and does not directly inhibit mTOR or other targets in vitro. Its activity lies in its utility as a building block for synthesizing active molecules. The final C26-linked rapamycin analogs synthesized from this intermediate are expected to retain the mTOR inhibitory activity of rapamycin, which has been extensively characterized. In vitro assays for mTOR inhibition include measurement of phosphorylation of downstream substrates such as p70S6 kinase (p70S6K) and 4E-BP1 in cell lysates by Western blotting, using purified mTOR complex in kinase activity assays, or using cell-based proliferation assays. Any biological activity observed in a final product derived from this intermediate should be attributed to the full conjugate, not to the intermediate itself. For research purposes, the intermediate is typically used as a chemical reagent without further biological characterization.
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| ln Vivo |
C57BL/6 mice.
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| Enzyme Assay |
There is no standard non-cellular enzyme assay for XL388-C2-amide-PEG9-NH2 hydrochloride, as it is a chemical intermediate without inherent biological activity. Instead, the intermediate is characterized by analytical chemistry methods: high-performance liquid chromatography (HPLC) for purity assessment (>95% by typical specifications); mass spectrometry (MS) to confirm molecular weight (C43H₆3ClFN₅O13S, exact mass ~944.50 g/mol); and nuclear magnetic resonance (NMR) spectroscopy to confirm structure (¹H NMR, ¹3C NMR, and 2D NMR correlation experiments). If the final synthesized product (a C26-linked rapamycin analog) is to be tested for mTOR binding affinity, a standard mTOR kinase assay can be performed using recombinant mTOR complex and an appropriate substrate. For example, an in vitro kinase assay would include purified mTOR, ATP, and a substrate peptide, and the reaction is monitored by measuring ADP production using a luminescent ADP detection kit or by detection of phosphorylated substrate by ELISA or Western blot. However, the intermediate itself is not used in such assays.
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| Cell Assay |
There is no standard in vitro cell-based assay for XL388-C2-amide-PEG9-NH2 hydrochloride itself, as it is a chemical intermediate. However, if the final conjugate (a C26-linked rapamycin analog) is to be tested, typical cell-based assays for mTOR inhibition are performed. Cells (e.g., HEK293, HeLa, MCF-7, or primary cells) are cultured in DMEM supplemented with 10% FBS at 37degC with 5% CO2. Cells are seeded in 6-well or 96-well plates and allowed to reach 70-80% confluence. They are then treated with varying concentrations of the final test compound (0.1-1000 nM) for 4-24 h. After treatment, cells are lysed, and the phosphorylation status of mTOR downstream targets (p70S6K at Thr389, 4E-BP1 at Thr37/46, and AKT at Ser473) is determined by Western blotting using phospho-specific antibodies. Cell proliferation is measured using the MTT assay or the CellTiter-Glo luminescent cell viability assay after 48-72 h of treatment. Flow cytometry can be used to assess cell cycle distribution (e.g., G1 arrest induced by mTOR inhibition). Cytotoxicity is evaluated using LDH release or propidium iodide staining. For the intermediate itself, no such assays are performed.
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| Animal Protocol |
In vivo animal studies for this intermediate are not performed because it is not a final therapeutic agent. If a final C26-linked rapamycin analog synthesized using this intermediate is to be tested in animals, typical protocols for rapalogs would apply. For example, female BALB/c nude mice bearing subcutaneous tumor xenografts (e.g., HCT116 colon cancer cells or other rapamycin-sensitive tumors) are used. When tumors reach ~100-200 mm3, animals are randomized to treatment groups (n = 5-10 per group). The test compound is formulated in a suitable vehicle (e.g., 5% DMSO, 5% Tween-80, 90% saline) and administered intraperitoneally (IP) or orally (by gavage) at doses of 1-20 mg/kg, once daily or every other day for 14-21 days. Tumor volume is measured every 2-3 days using calipers, and tumor growth inhibition (TGI) is calculated as the percentage reduction in tumor volume compared to vehicle control. Body weight is monitored as a measure of toxicity. At study termination, tumors are excised and processed for histology or Western blot analysis to confirm mTOR pathway inhibition (reduced p-p70S6K and p-4E-BP1). All animal studies require prior institutional animal ethics approval. The intermediate itself is not administered to animals.
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| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties of XL388-C2-amide-PEG9-NH2 hydrochloride itself have not been studied, as it is a chemical intermediate with no intended in vivo use. For the final C26-linked rapamycin analog synthesized from this intermediate, PK properties may vary depending on the specific structure of the conjugate. Rapamycin itself has poor aqueous solubility and low oral bioavailability, requiring specialized formulation (e.g., nanoparticle formulations or inclusion complexes). Rapamycin has a long elimination half-life (t1/2 ~ 60 h in humans), extensive tissue distribution, and is metabolized primarily by CYP3A4 in the liver. The introduction of a PEG9 linker and terminal NH2 group may alter solubility, tissue distribution, and elimination half-life compared to rapamycin, potentially increasing aqueous solubility and altering biodistribution. However, no specific PK data are available for any conjugate derived from this intermediate. This compound is for research use only; not for human administration.
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| Toxicity/Toxicokinetics |
Toxicological data for XL388-C2-amide-PEG9-NH2 hydrochloride are not available, as the compound is a chemical intermediate and is not intended for in vivo use. Standard safety precautions for handling organic compounds should be followed: wear appropriate personal protective equipment (PPE), including gloves, lab coat, and safety goggles. Avoid inhalation and contact with skin and eyes. Work should be performed in a well-ventilated area, preferably in a fume hood. The compound is intended for laboratory use only and should not be ingested or injected. If the final product derived from this intermediate is intended for in vivo use, a full toxicological evaluation would be required, including acute, subchronic, and chronic toxicity studies, genotoxicity assays, and reproductive/developmental toxicity studies, in accordance with regulatory guidelines. For research use only; not intended for human therapeutic or diagnostic use.
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| References |
[1]. Christopher Michael Semko, et al. C26-linked rapamycin analogs as mtor inhibitors. WO2019212991A1.
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| Additional Infomation |
XL388-C2-amide-PEG9-NH2 hydrochloride is not a drug and is not approved for any human use. It is a synthetic chemical intermediate used exclusively for research purposes in the synthesis of C26-linked rapamycin analogs. Its mechanism (in the context of the final product) relates to the synthesis of modified rapamycin derivatives that can be conjugated to other molecules for studying mTOR biology or for developing targeted therapies. The PEG9 linker improves hydrophilicity and reduces aggregation of the conjugate, while the terminal NH2 group allows for further conjugation (e.g., to biotin, fluorescent dyes, peptides, or antibodies). No clinical trials have been registered for this intermediate or for any final products derived from it. For research use only; not for diagnostic, therapeutic, or clinical applications in humans.
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| Molecular Formula |
C43H63CLFN5O13S
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| Related CAS # |
XL388-C2-amide-PEG9-NH2 TFA
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| Appearance |
Light yellow to yellow oil
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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: Please store this product in a sealed and protected environment, 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) |
DMSO :~250 mg/mL (~264.69 mM)
H2O :~100 mg/mL (~105.88 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 4.17 mg/mL (4.42 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 41.7 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 4.17 mg/mL (4.42 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 41.7 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 4.17 mg/mL (4.42 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 100 mg/mL (105.88 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
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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