MMAD (also known as Monomethyl auristatin D), an auristatin analog, is a novel and highly potent tubulin inhibitor/antimitotic agent that is often used as a warhead or a toxin payload in antibody drug conjugates (ADCs). Antibodies covalently linked to extremely powerful drugs through a range of conjugation technologies are known as antibody-drug conjugates (ADCs). When used as therapeutics, they combine the cytotoxic drug's capacity to kill cells with the exceptional specificity of antibodies, which allows for the differentiation between healthy and diseased tissue. With two US Food and Drug Administration-approved ADCs now on the market (Adcetris and Kadcyla) and about 40 more undergoing clinical evaluation, this potent and exciting class of targeted therapy has shown considerable promise in the treatment of various cancers. The majority of these ADCs, however, are heterogeneous mixtures, which can have significant pharmacokinetic effects and lead to a limited therapeutic window. Sophisticated site-specific conjugation technologies to link the drug and antibody are essential for ADCs to perform to the best of their abilities.
Physicochemical Properties
| Molecular Formula | C41H66N6O6S |
| Molecular Weight | 771.0643 |
| Exact Mass | 770.476 |
| Elemental Analysis | C, 63.87; H, 8.63; N, 10.90; O, 12.45; S, 4.16 |
| CAS # | 203849-91-6 |
| Related CAS # | MMAD-d8 |
| PubChem CID | 10723894 |
| Appearance | White to off-white solid powder |
| Density | 1.1±0.1 g/cm3 |
| Boiling Point | 906.1±65.0 °C at 760 mmHg |
| Flash Point | 501.8±34.3 °C |
| Vapour Pressure | 0.0±0.3 mmHg at 25°C |
| Index of Refraction | 1.537 |
| LogP | 5.82 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 9 |
| Rotatable Bond Count | 21 |
| Heavy Atom Count | 54 |
| Complexity | 1190 |
| Defined Atom Stereocenter Count | 9 |
| SMILES | S1C([H])=C([H])N=C1[C@]([H])(C([H])([H])C1C([H])=C([H])C([H])=C([H])C=1[H])N([H])C([C@]([H])(C([H])([H])[H])[C@]([H])([C@]1([H])C([H])([H])C([H])([H])C([H])([H])N1C(C([H])([H])[C@]([H])([C@]([H])([C@@]([H])(C([H])([H])[H])C([H])([H])C([H])([H])[H])N(C([H])([H])[H])C([C@]([H])(C([H])(C([H])([H])[H])C([H])([H])[H])N([H])C([C@]([H])(C([H])(C([H])([H])[H])C([H])([H])[H])N([H])C([H])([H])[H])=O)=O)OC([H])([H])[H])=O)OC([H])([H])[H])=O |
| InChi Key | BLUGYPPOFIHFJS-UUFHNPECSA-N |
| InChi Code | InChI=1S/C41H66N6O6S/c1-12-27(6)36(46(9)41(51)35(26(4)5)45-39(50)34(42-8)25(2)3)32(52-10)24-33(48)47-21-16-19-31(47)37(53-11)28(7)38(49)44-30(40-43-20-22-54-40)23-29-17-14-13-15-18-29/h13-15,17-18,20,22,25-28,30-32,34-37,42H,12,16,19,21,23-24H2,1-11H3,(H,44,49)(H,45,50)/t27-,28+,30-,31-,32+,34-,35-,36-,37+/m0/s1 |
| Chemical Name | (2S)-N-[(2S)-1-[[(3R,4S,5S)-3-methoxy-1-[(2S)-2-[(1R,2R)-1-methoxy-2-methyl-3-oxo-3-[[(1S)-2-phenyl-1-(1,3-thiazol-2-yl)ethyl]amino]propyl]pyrrolidin-1-yl]-5-methyl-1-oxoheptan-4-yl]-methylamino]-3-methyl-1-oxobutan-2-yl]-3-methyl-2-(methylamino)butanamide |
| Synonyms | Monomethylauristatin D; MMAD; Demethyldolastatin 10; Monomethyl Dolastatin 10 |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.(2). This product is not stable in solution, please use freshly prepared working solution for optimal results. |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets | Auristatin |
| ln Vitro | MMAD (Monomethyl Dolastatin 10) is joined via a steady oxime-ligation procedure to produce a number of nearly homogenous drug-to-antibody conjugates (ADCs) with an approximate drug-to-antibody ratio of 2.0. The resultant conjugates exhibit strong in vitro cytotoxic activity against HER2+ cancer cells along with good pharmacokinetic characteristics. Site-specific unnatural amino acid-based ADCs are demonstrated to exhibit higher in vitro cytotoxicity in comparison to ADCs made by cysteine alkylation after native interchain disulfide reduction[1]. |
| ln Vivo | In rodents, the resultant antibody-drug conjugates (ADCs) show total tumor regression. Additionally, they have a better toxicology profile in rats [1]. |
| References |
[1]. Recent advances in the construction of antibody-drug conjugates. Nat Chem. 2016 Feb;8(2):114-9. |
Solubility Data
| Solubility (In Vitro) | DMSO: ~24.5 mg/mL (~31.8 mM) |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.2969 mL | 6.4846 mL | 12.9692 mL | |
| 5 mM | 0.2594 mL | 1.2969 mL | 2.5938 mL | |
| 10 mM | 0.1297 mL | 0.6485 mL | 1.2969 mL |