AG-205
Based on 1 Customer Validation
AG-205 is a progesterone receptor membrane component 1 (PGRMC1) antagonist and CGT inhibitor, with an IC50 of 50 μM against rat CGT. AG-205 exhibits antimitotic, antimigratory and anti-invasive activities. AG-205 increases the expression of genes encoding cholesterol biosynthesis pathway or steroidogenic enzymes. AG-205 promotes the regulation of cell cycle by apoptosis and reduces the migratory and invasive capacities of ovarian and breast cancer cells. AG-205 can be used in research related to renal cancer and breast cancer.
Nur für Forschungszwecke. Wir verkaufen nicht an Patienten.
- Reinheit : 99%
- CAS. Nr.: 1375078-57-1
- Formel: C22H23ClN6OS
- Molecular Weight:454.98
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Speicherung:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biologische Aktivität
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Chemical Information
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Lösungsmittel & Löslichkeit
- Protokoll
- Reinheit & Dokumentation
- Verweise
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Help & FAQs
Biologische Aktivität
Beschreibung
In Vitro
AG-205 (10-40 μM; 24 h) significantly increases the survival rate of SMKT-R3 human renal cancer cells[1].
AG-205 (2-10 μM) specifically reduces sulfatide levels in SMKT-R3 human renal cancer cells, with no effect on major non-sphingolipids[1].
AG-205 (10 μM; 6 h) specifically inhibits the synthesis of CGT-dependent galactolipids (GalC, MGDG) and sulfatides in CHO-CGT and CHO-Sulf cells expressing galactosylceramide synthase, while promoting the synthesis of GlcC[1].
AG-205 (10 μM; 48 h) inhibits the synthesis of GalC and MGDG and promotes the synthesis of GlcC in CHO-CGT cells, and this effect is independent of the expression of PGRMC1 or PGRMC2[1].
AG-205 (50 μM; 30 min) significantly inhibits the in vitro activity of UDP-galactose:ceramide galactosyltransferase (CGT) derived from CHO-CGT cells[1].
Pretreatment with AG-205 (50 μM; 24 h) abolishes the anti-apoptotic effect of progesterone against H2O2-induced cell death in human granulosa/luteal cells[2].
AG-205 (50 μM; overnight) reduces the nuclear localization level of PGRMC1 in human granulosa/luteal cells, with most PGRMC1 retained in the cytoplasm[2].
AG-205 (50 μM; overnight) increases the cytoplasmic level of HRK protein in human granulosa/luteal cells[2].
AG-205 (50 μM; overnight) disrupts the colocalization of cytochrome c with mitochondria in human granulosa/luteal cells, but does not alter the total level of cytochrome c in the cells[2].
AG-205 (50 μM; overnight) enhances the interaction between PGRMC1 and PGRMC2 in human granulosa/luteal cells, as evidenced by a 2.3-fold increase in PLA dots per cell[2].
AG-205 (50 μM; overnight) increases the expression level of Hrk mRNA by 6-8 folds in human granulosa/luteal cells, while slightly elevating the mRNA expression levels of several anti-apoptotic genes by 1.5-3 folds[2].
AG-205 (15 μM; 32 h) significantly upregulates the mRNA expression levels of genes encoding cholesterol biosynthetic enzymes, the sterol regulatory factor INSIG1, and steroidogenic enzymes in human endometrial T-HESC and HEC-1A cell lines[3].
AG-205 (15 μM; 80 h) upregulates the mRNA expression of HSD17B7, MSMO1 and INSIG1 in human endometrial T-HESC and HEC-1A cell lines, and this process is independent of PGRMC1 expression[3].
AG-205 (15 μM; 80 h) upregulates the mRNA expression of HSD17B7, MSMO1, and INSIG1 in human endometrial T-HESC and HEC-1A cell lines, and this process is independent of the expression of all four members of the MAPR family (PGRMC1, PGRMC2, NENF, CYB5D2)[3].
AG-205 (15 μM; 32 h) does not alter the mRNA or protein expression levels of PGRMC1 in the human endometrial T-HESC and HEC-1A cell lines, nor does it change the subcellular localization of PGRMC1 in these cell lines[3].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:human granulosa/luteal cells
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Concentration:50 μM
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Incubation Time:24 h (pretreatment)
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Result:Eliminated progesterone's ability to attenuate H2O2-induced apoptosis, resulting in a dead cell/total cell ratio equivalent to H2O2 treatment alone.
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Cell Line:human granulosa/luteal cells
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Concentration:50 μM
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Incubation Time:overnight
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Result:Reduced nuclear localization of PGRMC1; 31 ± 10% of cells showed nuclear PGRMC1 staining, compared to 67 ± 8% in controls.
Prominently localized PGRMC1 to the cytoplasm in treated cells.\nIncreased levels of HRK protein in the cytoplasm of treated cells, visible as enhanced green fluorescence compared to controls.\nAltered the subcellular localization of cytochrome c, with a portion of cytochrome c no longer co-localizing with mitochondria (visible as diffuse green fluorescence in the cytoplasm) instead of being restricted to punctate mitochondrial foci.
Left total cellular cytochrome c levels unchanged relative to controls.
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Cell Line:human granulosa/luteal cells
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Concentration:50 μM
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Incubation Time:overnight
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Result:Increased abundance of the ~27 kDa monomeric form of PGRMC1.
Decreased abundance of the higher molecular weight forms ≥50 kDa (dimers/oligomers) of PGRMC1.
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Cell Line:human granulosa/luteal cells
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Concentration:50 μM
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Incubation Time:overnight
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Result:Induced a 6 to 8 fold increase in Harakiri (Hrk) mRNA levels.
Induced 1.5 to 3 fold increases in mRNA levels of Bag1, Bcl2A1, Birc3, Mcl1, and XIAP.
Induced a small increase in Casp4 mRNA.
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Cell Line:human endometrial T-HESC, HEC-1A
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Concentration:5 μM, 15 μM, 30 μM, 45 μM
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Incubation Time:24 h, 32 h, 48 h
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Result:Had no effect on cell viability in either cell line for up to 48 h when used at 5 μM or 15 μM.
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Cell Line:human endometrial T-HESC, HEC-1A
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Concentration:15 μM
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Incubation Time:32 h
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Result:Significantly upregulated mRNA expression of multiple enzymes involved in cholesterol biosynthesis, including ACAT2, AACS, HMGCS1, HMGCR, MVK, PMVK, MVD, IDI1, FDPS, FDFT1, SQLE, LSS, CYP51A1, TM7SF2, MSMO1, NSDHL, HSD17B7, EBP, SC5D, DHCR7, and DHCR24, in both cell lines.
Upregulated mRNA expression of INSIG1 (a sterol-sensitive regulator) and steroidogenesis-related enzymes AKR1C1 and HSD17B14 in both cell lines.
Confirmed significant upregulation of HSD17B7, MSMO1, and INSIG1 in both cell lines via qPCR.
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Cell Line:human endometrial T-HESC, HEC-1A (with prior siRNA-mediated knockdown of PGRMC1)
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Concentration:15 μM
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Incubation Time:32 h (following 48 h siRNA transfection)
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Result:Induced a ~3 to 4-fold mean increase in HSD17B7, MSMO1, and INSIG1 mRNA expression in HEC-1A cells, and a ~4 to 6-fold mean increase in T-HESC cells.
Maintained identical upregulation in cells transfected with PGRMC1-targeting siRNA or control siRNA.
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Cell Line:human endometrial T-HESC, HEC-1A (with prior siRNA-mediated knockdown of all four MAPR family members: PGRMC1, PGRMC2, NENF, CYB5D2)
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Concentration:15 μM
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Incubation Time:32 h (following 48 h siRNA transfection)
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Result:Significantly upregulated mRNA expression of HSD17B7, MSMO1, and INSIG1 in both cell lines.
Maintained upregulation at the same magnitude in cells transfected with the MAPR-targeting siRNA mixture as in control siRNA-transfected cells.
Chemical Information
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CAS. Nr. 1375078-57-1
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Appearance Solid
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Molecular Weight 454.98
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Formel C22H23ClN6OS
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Color White to off-white
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SMILES
CC1=CC2=C(C=C1)N([C@@]3([H])[C@]2([H])CN(CC3)C)C(CSC4=NN=NN4C5=CC=C(C=C5)Cl)=O
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Versand
Room temperature in continental US; may vary elsewhere.
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Speicherung
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Lösungsmittel & Löslichkeit
In Vitro:
DMSO : 10 mg/mL (21.98 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Konzentration (Stammlösung) × Volumen (Stammlösung) = Konzentration (Ziellösung) × Volumen (Ziellösung)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: 1 mg/mL (2.20 mM); Suspended solution; Need ultrasonic
This protocol yields a suspended solution of 1 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (10.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protokoll
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RNA extraction experimental
By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.
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Apoptosis
Apoptosis, also called programmed cell death, is generally characterized by distinct morphological characteristics.
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TUNEL staining for apoptotic DNA fragmentation
TUNEL staining detects DNA strand breaks by using terminal deoxynucleotidyl transferase to add labeled nucleotides to exposed 3′-OH DNA termini, generating either microscopic staining in fixed cells or tissue sections, or fluorescence/cytometric signal in cell suspensions. TUNEL positivity reflects DNA fragmentation but should not be interpreted alone as definitive apoptosis, because TUNEL can also label necrotic, autolytic, mechanically damaged, or DNA-repair-associated DNA breaks.
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Flow cytometric DNA-content cell-cycle staining
Flow cytometric DNA-content cell-cycle staining measures the fluorescence intensity of DNA-bound fluorochromes in single cells or nuclei to estimate DNA content distributions, allowing assignment of populations to G0/G1, S, and G2/M phases by DNA histogram deconvolution. Propidium iodide (PI) intercalates into DNA, and PI fluorescence is proportional to cellular DNA content when staining is performed under conditions that make DNA accessible and minimize non-DNA signal. Cells with G2/M DNA content are expected to show approximately twice the fluorescence intensity of G0/G1 cells, while S-phase cells occupy intermediate fluorescence values. PI-based DNA-content analysis can also detect cells with fractional DNA content, often reported as sub-G1, when DNA fragmentation and extraction during staining reduce retained DNA signal in apoptotic cells. DAPI is an alternative DNA fluorochrome for univariate DNA-content analysis, while bivariate approaches combining DNA content with proliferation
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Annexin V plus membrane-impermeant dye apoptosis staining
Annexin V-based apoptosis assays rely on the detection of phosphatidylserine (PS) externalization from the inner leaflet of the plasma membrane to the outer leaflet, an early biochemical hallmark of apoptosis. Fluorescently labeled Annexin V binds PS in a calcium-dependent manner, enabling identification of early apoptotic cells by flow cytometry or fluorescence microscopy. When combined with a membrane-impermeant DNA-binding dye (e. g. , propidium iodide), this approach allows discrimination between viable (Annexin V−/dye−), early apoptotic (Annexin V+/dye−), and late apoptotic or necrotic (Annexin V+/dye+) cell populations by assessing membrane integrity and PS exposure.
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BrdU Incorporation Assay
Bromodeoxyuridine (BrdU) incorporation assay is based on the principle that BrdU, a thymidine analog, is incorporated into newly synthesized DNA during the S phase of the cell cycle, thereby serving as a marker of DNA replication and cellular proliferation. Incorporated BrdU can be detected using anti-BrdU antibodies following DNA denaturation, enabling visualization or quantification of proliferating cells through immunochemical detection methods such as immunofluorescence or immunohistochemistry.
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Apoptosis Solutions
Apoptosis is a regulated, generally non-lytic cell-death pathway that removes unwanted, damaged, infected, or abnormal cells through coordinated morphological changes, caspase activation, DNA fragmentation, and membrane remodeling. The intrinsic apoptosis pathway is controlled mainly by mitochondrial outer membrane permeabilization, BCL-2 family proteins, cytochrome c release, apoptosome formation, caspase-9 activation, and downstream executioner caspase-3/7 activation. The extrinsic apoptosis pathway is initiated by death receptors such as Fas, TNFR, and TRAIL receptors, which recruit adaptor proteins and activate caspase-8 before engaging executioner caspases or mitochondrial amplification through BID cleavage. Apoptosis is linked to many phenotypes, including cancer cell killing, tissue homeostasis, immune regulation, neurodegeneration, infection response, and treatment-induced cytotoxicity; unresolved questions include how apoptosis interacts with necroptosis, pyroptosis, ferroptos
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Protocol for Cell Cycle
Cell-cycle analysis by flow cytometry measures DNA content in single cells to estimate the fraction of cells in G0/G1, S, and G2/M phases. Propidium iodide intercalates into DNA, and after RNA removal with RNase, fluorescence intensity reflects cellular DNA content: 2N cells are assigned to G0/G1, cells between 2N and 4N to S phase, and 4N cells to G2/M. DNA-content analysis alone cannot reliably separate G0 from G1 or G2 from M. Ki-67 can distinguish quiescent G0 cells from cycling cells, EdU or BrdU incorporation marks active DNA synthesis in S phase, and phospho-histone H3 staining identifies mitotic cells within the 4N population.
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Breast Cancer Modeling
Breast cancer is a heterogeneous cancer, and it has been distinguished into four subtypes: luminal A, luminal B, HER2-positive and basal-like. Molecular mutations, epigenetic alterations, hormone exposure and immune microenvironment are related to the progression of breast cancer.
Reinheit & Dokumentation
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Data Sheet (290 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
Verweise
[1]. Wang-Eckhardt L, et al. The PGRMC1 Antagonist AG-205 Inhibits Synthesis of Galactosylceramide and Sulfatide. Cells. 2021;10(12):3520. Published 2021 Dec 13. [Content Brief]
[3]. Thieffry C, et al. AG-205 Upregulates Enzymes Involved in Cholesterol Biosynthesis and Steroidogenesis in Human Endometrial Cells Independently of PGRMC1 and Related MAPR Proteins. Biomolecules. 2021;11(10):1472. Published 2021 Oct 6. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
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| DMSO | 1 mM | 2.1979 mL | 10.9895 mL | 21.9790 mL | 54.9475 mL |
| 5 mM | 0.4396 mL | 2.1979 mL | 4.3958 mL | 10.9895 mL | |
| 10 mM | 0.2198 mL | 1.0989 mL | 2.1979 mL | 5.4947 mL | |
| 15 mM | 0.1465 mL | 0.7326 mL | 1.4653 mL | 3.6632 mL | |
| 20 mM | 0.1099 mL | 0.5495 mL | 1.0989 mL | 2.7474 mL |
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