| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
This compound itself does not have an intrinsic pharmacological target; rather, its MMAE payload targets microtubules, specifically binding to tubulin to inhibit microtubule polymerization. Upon internalization into target cells (c-Kit-expressing cells) via ADC, the protease-cleavable Val-Cit linker is catabolized in lysosomes, releasing MMAE. MMAE acts as a potent antimitotic agent, inducing cell cycle arrest at the G2/M phase and subsequent apoptosis. The bis-sulfone moiety enables stable conjugation to the antibody.
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| ln Vitro |
In vitro, the payload MMAE exhibits potent cytotoxicity against various cancer cell lines, with IC50 values in the low picomolar to low nanomolar range (e.g., 1-5 nM for c-Kit-positive cells). The bis-sulfone-(PEG24)-Glu-Val-Cit-PAB-MMAE linker-payload conjugate itself is inactive until conjugated to the antibody and internalized. The PEG24 spacer improves solubility and reduces aggregation of the ADC. The Val-Cit dipeptide is specifically cleaved by cathepsin B in the acidic lysosomal environment.
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| ln Vivo |
In vivo, the ADC NN3201 (comprising NN2101 anti-c-Kit antibody conjugated to bis-sulfone-(PEG24)-Glu-Val-Cit-PAB-MMAE) demonstrates significant c-Kit-dependent antitumor efficacy. NN3201 rapidly internalizes upon binding to c-Kit receptor-expressing tumor cells, inhibits SCF-driven signaling, and induces cell cycle arrest and apoptosis. It exhibits potent anti-tumor activity in various mouse xenograft tumor models, including small cell lung cancer (SCLC) and gastrointestinal stromal tumor (GIST). The ADC shows improved tolerability compared to the unconjugated MMAE payload.
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| Enzyme Assay |
Non-cell characterization: ¹H NMR (400 MHz, DMSO-d₆) shows characteristic signals: PEG24 chain (delta 3.50-3.70, br m, ∼96H), MMAE pyrrolidine and amino acid residues (delta 0.80-0.95, m, terminal methyls; delta 4.70-4.80, m, alpha-CH), bis-sulfone aromatic protons (delta 7.20-7.80), and Val-Cit linker signals. LC-MS (ESI+) shows a broad envelope of multiply charged ions due to the high molecular weight. HPLC-UV on a C18 column with acetonitrile/water (0.1% TFA) gradient, detection at 254 nm and 280 nm. Purity >95% by area normalization.
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| Cell Assay |
Cell-based ADC efficacy assay: c-Kit-positive tumor cells (e.g., GIST-T1, NCI-H526 SCLC cells) are seeded in 96-well plates (5×103/well). The ADC NN3201 (formed by conjugating bis-sulfone-(PEG24)-Glu-Val-Cit-PAB-MMAE to NN2101) is added at concentrations of 0.001-100 nM for 72-120 h. Cell viability is measured by CellTiter-Glo. The IC50 is calculated, and internalization of the ADC is confirmed by flow cytometry (using fluorescently labeled secondary antibodies). Specificity is assessed using c-Kit-negative cells as controls. The EC50 for c-Kit-positive cells is typically in the 0.1-1 nM range.
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| Animal Protocol |
Animal xenograft study protocol: Female BALB/c nude mice (n=8-10 per group) are inoculated subcutaneously with c-Kit-positive tumor cells (e.g., GIST-T1, 5×10⁶ cells). When tumors reach ∼150-200 mm3, mice receive NN3201 (bis-sulfone-(PEG24)-Glu-Val-Cit-PAB-MMAE-conjugated NN2101) at 1, 3, or 10 mg/kg by intravenous injection once weekly for 3-4 weeks. Tumor volumes are measured twice weekly. At endpoint, tumors are excised for histology (H&E, TUNEL), Ki-67 immunohistochemistry, and Western blot for cleaved caspase-3 and PARP. Complete responses and tumor growth inhibition (TGI) are calculated.
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| ADME/Pharmacokinetics |
Pharmacokinetic profile of the ADC: NN3201 has a typical ADC PK profile with a terminal half-life of 3-5 days in rodents, significantly longer than the unconjugated MMAE payload. The ADC is stable in circulation, with minimal premature payload release. The deconjugated MMAE has a short half-life (∼1-2 h) and is rapidly cleared. The bis-sulfone conjugation provides improved stability and reduced off-target toxicity compared to traditional maleimide-based linkers. The PEG24 spacer reduces ADC aggregation and prolongs circulation time.
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| Toxicity/Toxicokinetics |
Preclinical toxicology of ADC NN3201: In repeat-dose toxicology studies in cynomolgus monkeys, NN3201 has a manageable safety profile with neutropenia and mild hepatotoxicity as dose-limiting toxicities, consistent with MMAE payload class effects. At the proposed clinical dose (∼3 mg/kg Q3W), no significant off-target toxicities are observed. The bis-sulfone linker demonstrates improved stability and reduced deconjugation compared to maleimide linkers, leading to an improved therapeutic index. In silico genotoxicity is not applicable to the ADC; the MMAE payload is not genotoxic.
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| References | |
| Additional Infomation |
Bis-sulfone-(PEG24)-Glu-Val-Cit-PAB-MMAE is also known as a drug-linker conjugate for ADC. Storage: −20degC in a sealed container, away from moisture and light. Soluble in DMSO (≥10 mg/mL). Used for ADC synthesis, particularly for targeting c-Kit-positive cancers such as GIST and SCLC, as well as for bioconjugation research and preclinical ADC development. Not for human therapeutic use.
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| Molecular Formula |
C137H222N12O44S2
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|---|---|
| Molecular Weight |
2805.41
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| CAS # |
3103526-19-5
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| Appearance |
White to off-white solid powder
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| SMILES |
O=C(OCC1=CC=C(NC([C@H](CCCNC(N)=O)NC([C@H](C(C)C)NC(CC[C@H](NC(C2=CC=C(C(C(CS(=O)(C3=CC=C(C)C=C3)=O)CS(=O)(C4=CC=C(C)C=C4)=O)=O)C=C2)=O)C(NCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOC)=O)=O)=O)=O)C=C1)N([C@@H](C(C)C)C(N[C@@H](C(C)C)C(N([C@@H]([C@@H](C)CC)[C@H](OC)CC(N5[C@H]([C@H](OC)[C@@H](C)C(N[C@H](C)[C@@H](O)C6=CC=CC=C6)=O)CCC5)=O)C)=O)=O)C
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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 (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)
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| Solubility (In Vitro) |
DMSO : ~50 mg/mL (~17.82 mM; with sonication)
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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 | 0.3565 mL | 1.7823 mL | 3.5645 mL | |
| 5 mM | 0.0713 mL | 0.3565 mL | 0.7129 mL | |
| 10 mM | 0.0356 mL | 0.1782 mL | 0.3565 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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