 Inhibitor Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C13H15NO6S |
| Molecular Weight | 313.3263 |
| Exact Mass | 313.062 |
| CAS # | 192203-60-4 |
| PubChem CID | 9796834 |
| Appearance | White to off-white solid powder |
| Density | 1.43 |
| LogP | 2.019 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 8 |
| Heavy Atom Count | 21 |
| Complexity | 381 |
| Defined Atom Stereocenter Count | 1 |
| SMILES | COC1=CC=C(C=C1)SC(=O)N[C@@H](CCC(=O)O)C(=O)O |
| InChi Key | OBCGYIKABKFIQB-JTQLQIEISA-N |
| InChi Code | InChI=1S/C13H15NO6S/c1-20-8-2-4-9(5-3-8)21-13(19)14-10(12(17)18)6-7-11(15)16/h2-5,10H,6-7H2,1H3,(H,14,19)(H,15,16)(H,17,18)/t10-/m0/s1 |
| Chemical Name | (2S)-2-[(4-methoxyphenyl)sulfanylcarbonylamino]pentanedioic acid |
| 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 |
| 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 |
1. Carboxypeptidase G2 (CPG2, a folate-degrading metalloenzyme, Ki = 0.8 nM for the lead Carboxypeptidase G2 (CPG2) Inhibitor; IC50 = 2.1 nM for recombinant CPG2 enzyme activity inhibition) [1] |
| ln Vitro |
1. CPG2 enzyme inhibitory activity: Carboxypeptidase G2 (CPG2) Inhibitor (lead compound in the study) exhibited potent, competitive inhibition of recombinant bacterial CPG2 enzyme; it had a Ki value of 0.8 nM and an IC50 of 2.1 nM for the hydrolysis of the substrate methotrexate (MTX) by CPG2. The inhibitor showed high selectivity for CPG2, with no significant inhibition of other carboxypeptidases (e.g., carboxypeptidase A, carboxypeptidase B) at concentrations up to 1 μM (residual enzyme activity > 90% for these off-target enzymes) [1] 2. Folate protection effect: In in vitro folate degradation assays, Carboxypeptidase G2 (CPG2) Inhibitor (10 nM) prevented CPG2-mediated cleavage of folic acid and its analogs; at this concentration, the residual folate level in the reaction system was increased by 89% compared with the CPG2-alone control group, confirming its ability to block CPG2-dependent folate catabolism [1] |
| Enzyme Assay |
1. Recombinant CPG2 enzyme activity inhibition assay: The assay was established in a buffer system containing purified recombinant CPG2 enzyme, folate-based substrate (methotrexate), and serial dilutions of Carboxypeptidase G2 (CPG2) Inhibitor (0.1 nM–10 μM). The reaction was initiated by adding the enzyme and incubated at 37℃ for 20 min, then terminated by adding a stop solution that quenched the enzymatic reaction. The remaining intact substrate was quantified using a UV-visible spectrophotometer at a wavelength of 302 nm (specific for methotrexate). Enzyme activity was calculated based on the substrate consumption rate, and the Ki and IC50 values were derived by fitting the dose-response curves with a competitive inhibition model [1] 2. Carboxypeptidase selectivity assay: A panel of purified carboxypeptidases (CPG2, carboxypeptidase A, carboxypeptidase B) was incubated with Carboxypeptidase G2 (CPG2) Inhibitor (concentrations up to 1 μM) and their respective specific substrates in buffer systems optimized for each enzyme. The enzymatic reaction was carried out at 37℃ for 30 min, and substrate hydrolysis was detected using substrate-specific spectrophotometric or fluorometric methods. Residual enzyme activity was calculated relative to the vehicle control to evaluate the inhibitor’s target selectivity [1] |
| References |
[1]. Novel Inhibitors of Carboxypeptidase G2 (CPG2): Potential Use in Antibody-Directed Enzyme Prodrug Therapy. Journal of Medicinal Chemistry (1999), 42(6), 951-956. [2]. Attenuated Salmonella targets prodrug activating enzyme carboxypeptidase G2 to mouse melanoma and human breast and colon carcinomas for effective suicide gene therapy. Clin Cancer Res. 2008 Jul 1;14(13):4259-66. [3]. Suicide gene therapy of human colon carcinoma xenografts using an armed oncolytic adenovirus expressing carboxypeptidase G2. Cancer Res. 2007 May 15;67(10):4949-55. [4]. Systemic gene-directed enzyme prodrug therapy of hepatocellular carcinoma using a targeted adenovirus armed with carboxypeptidase G2. Cancer Res. 2005 Jun 15;65(12):5003-8. |
| Additional Infomation |
1. Carboxypeptidase G2 (CPG2) Inhibitor is a novel, small-molecule metalloenzyme inhibitor designed to target bacterial and recombinant CPG2, with a core structure containing a zinc-binding moiety that chelates the active-site metal ion of CPG2 and a hydrophobic scaffold that occupies the enzyme’s substrate-binding pocket [1] 2. The inhibition mechanism of Carboxypeptidase G2 (CPG2) Inhibitor is competitive binding to CPG2’s active site: its zinc-binding group coordinates with the Zn²⁺ ion in CPG2’s catalytic center, while the adjacent hydrophobic region blocks the access of folate substrates (e.g., methotrexate) to the active site, thereby inhibiting CPG2-mediated folate degradation [1] 3. Carboxypeptidase G2 (CPG2) Inhibitor was developed for potential use in antibody-directed enzyme prodrug therapy (ADEPT): it can be used to modulate CPG2 activity in ADEPT systems, preventing off-target folate degradation and reducing systemic toxicity of CPG2-based prodrug therapies while maintaining localized prodrug activation at tumor sites [1] |
Solubility Data
| Solubility (In Vitro) | H2O : ~1.43 mg/mL (~4.56 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 | 3.1915 mL | 15.9576 mL | 31.9152 mL | |
| 5 mM | 0.6383 mL | 3.1915 mL | 6.3830 mL | |
| 10 mM | 0.3192 mL | 1.5958 mL | 3.1915 mL |