| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
Mal-GGFG-PAB-MMAE itself has no therapeutic target; it is a pro-drug. Its mechanism is triggered after conjugation to an antibody. The antibody targets a specific tumor antigen, and after internalization into the cancer cell, the linker is cleaved by lysosomal proteases (cathepsin B), releasing the active payload MMAE. MMAE then targets tubulin, inhibiting its polymerization, which leads to G2/M cell cycle arrest and apoptosis. The GGFG tetrapeptide sequence is specifically recognized and cleaved by cathepsin B.
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| ln Vitro |
The activity of Mal-GGFG-PAB-MMAE is not evaluated as a standalone molecule in free drug assays. As an ADC payload-linker, its in vitro activity is determined by the specificity of the antibody it is conjugated to. In ADC form, it exhibits potent and selective cytotoxicity against target antigen-positive cancer cells in the low nanomolar to picomolar range, with a high selectivity index (SI) over antigen-negative cells. The released MMAE has an IC50 of 0.1-10 nM against various cancer cell lines in standard proliferation assays.
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| ln Vivo |
ADCs synthesized using Mal-GGFG-PAB-MMAE show potent in vivo efficacy in xenograft mouse models. For example, an anti-HER2 ADC carrying this payload-linker induces tumor regression at doses as low as 1-10 mg/kg. The 'bystander effect' is observed due to the membrane permeability of the released MMAE, allowing it to diffuse into and kill neighboring antigen-negative tumor cells. The hydrophilic GGFG linker is designed for high plasma stability, minimizing premature payload release and systemic toxicity.
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| Enzyme Assay |
In vitro cathepsin B cleavage assay: The ADC is incubated with purified cathepsin B in cleavage buffer (50 mM sodium acetate, pH 5.0, 2 mM DTT) at 37degC for 1-4 hours. The release of MMAE is quantified by LC-MS/MS, monitoring the specific mass transition for MMAE. The rate of cleavage (pmol/min/ug enzyme) is calculated. This assay confirms the linker's cleavability by lysosomal proteases. The maleimide group's reactivity with thiols is confirmed by reacting it with cysteine-containing peptides and analyzing the adduct by mass spectrometry.
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| Cell Assay |
In vitro ADC cytotoxicity assay: Target antigen-positive and -negative cancer cells are seeded in 96-well plates (5,000 cells/well) and treated with serial dilutions (0.001-1000 ng/mL) of the ADC (conjugated to Mal-GGFG-PAB-MMAE) for 96-120 hours. Cell viability is measured by CellTiter-Glo. The IC50 for target-positive cells is calculated, and the selectivity index is determined. The bystander effect is assessed in co-culture assays with a mixture of antigen-positive and -negative cells; the killing of antigen-negative cells is measured by flow cytometry.
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| Animal Protocol |
Xenograft mouse model: Female BALB/c nude mice (6-8 wk, n=8/group) bearing subcutaneous target-positive tumor xenografts are used. When tumors reach ~100-200 mm3, the ADC is administered intravenously (tail vein) at doses of 1-10 mg/kg, once weekly for 2-3 weeks. Tumor volume is measured every 2-3 days with calipers. Complete regression is often observed at the highest doses. Pharmacodynamic analysis includes measurement of mitotic biomarkers (phospho-histone H3) and cleaved caspase-3 in tumor lysates. Body weight is monitored as a primary indicator of toxicity.
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| ADME/Pharmacokinetics |
The PK profile of the ADC (the final conjugate) is determined by the antibody, with a long half-life (days to weeks). The stability of the maleimide-thiol linkage is crucial. The hydrophilic GGFG linker reduces aggregation and improves plasma stability. The released MMAE (as the active catabolite) has a short half-life (< 1 h). The drug-to-antibody ratio (DAR) is a critical quality attribute, typically analyzed by hydrophobic interaction chromatography (HIC) or mass spectrometry. For research use, the compound is stored at -80degC.
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| Toxicity/Toxicokinetics |
The major toxicity of ADCs containing MMAE is often dose-limiting neutropenia and peripheral neuropathy, which are class-effects of tubulin inhibitors. The stability of the linker is critical; premature release of MMAE in circulation can cause increased systemic toxicity. Mal-GGFG-PAB-MMAE is for research use only and is not a drug product. Standard safety precautions for handling potent cytotoxic compounds should be followed.
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| References | |
| Additional Infomation |
Mal-GGFG-PAB-MMAE (CAS# 3055607-18-3) is a research-grade drug-linker conjugate for the site-specific synthesis of antibody-drug conjugates (ADCs). It utilizes a cathepsin B-cleavable peptide linker (GGFG) to attach the potent anti-mitotic agent MMAE to a thiolated antibody. It is not an FDA-approved drug and is strictly a research reagent. Storage: Powder at -80degC for 2 years, -20degC for 1 year; In solvent at -80degC for 6 months, -20degC for 1 month.
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| Molecular Formula |
C69H97N11O16
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|---|---|
| Molecular Weight |
1336.57
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| Exact Mass |
1335.711
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| CAS # |
3055607-18-3
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| PubChem CID |
172677432
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
8
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| Rotatable Bond Count |
37
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| Heavy Atom Count |
96
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| Complexity |
2640
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| Defined Atom Stereocenter Count |
11
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| SMILES |
CC[C@H](C)[C@@H]([C@@H](CC(=O)N1CCC[C@H]1[C@@H]([C@@H](C)C(=O)N[C@H](C)[C@H](C2=CC=CC=C2)O)OC)OC)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)N(C)C(=O)OCC3=CC=C(C=C3)NC(=O)CNC(=O)[C@H](CC4=CC=CC=C4)NC(=O)CNC(=O)CNC(=O)CCN5C(=O)C=CC5=O
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| InChi Key |
KIPVTXGIYUVJSR-ZKMHZRHYSA-N
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| InChi Code |
InChI=1S/C69H97N11O16/c1-13-43(6)62(52(94-11)36-59(87)79-33-20-25-51(79)64(95-12)44(7)65(89)73-45(8)63(88)48-23-18-15-19-24-48)77(9)68(92)60(41(2)3)76-67(91)61(42(4)5)78(10)69(93)96-40-47-26-28-49(29-27-47)74-55(83)39-72-66(90)50(35-46-21-16-14-17-22-46)75-56(84)38-71-54(82)37-70-53(81)32-34-80-57(85)30-31-58(80)86/h14-19,21-24,26-31,41-45,50-52,60-64,88H,13,20,25,32-40H2,1-12H3,(H,70,81)(H,71,82)(H,72,90)(H,73,89)(H,74,83)(H,75,84)(H,76,91)/t43-,44+,45+,50-,51-,52+,60-,61-,62-,63+,64+/m0/s1
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| Chemical Name |
[4-[[2-[[(2S)-2-[[2-[[2-[3-(2,5-dioxopyrrol-1-yl)propanoylamino]acetyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]acetyl]amino]phenyl]methyl N-[(2S)-1-[[(2S)-1-[[(3R,4S,5S)-1-[(2S)-2-[(1R,2R)-3-[[(1S,2R)-1-hydroxy-1-phenylpropan-2-yl]amino]-1-methoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl]-3-methoxy-5-methyl-1-oxoheptan-4-yl]-methylamino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]-N-methylcarbamate
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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 : ≥ 100 mg/mL (~74.82 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (1.87 mM)(saturation unknown) in 10% DMSO 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution and add it to 900 μL corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.7482 mL | 3.7409 mL | 7.4818 mL | |
| 5 mM | 0.1496 mL | 0.7482 mL | 1.4964 mL | |
| 10 mM | 0.0748 mL | 0.3741 mL | 0.7482 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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