| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
Serine hydroxymethyltransferase 1 and 2 (SHMT1 and SHMT2). These are pyridoxal 5'-phosphate (PLP)-dependent enzymes that catalyze the reversible conversion of serine and tetrahydrofolate (THF) to glycine and 5,10-methylenetetrahydrofolate, a key step in one-carbon metabolism. SHMT1 is primarily cytosolic, and SHMT2 is mitochondrial. SHIN1 is a folate-competitive inhibitor that targets both isoforms.
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| ln Vitro |
In biochemical assays, (-)-SHIN1 inhibits human SHMT1 and SHMT2 with IC50 values of 5 nM and 13 nM, respectively. It is a folate-competitive inhibitor that potently engages the cellular targets. At concentrations below 100 nM, it impairs cell growth due to its potent engagement of cellular SHMT1, but the (-) enantiomer is inactive compared to (+)-SHIN1. In HCT-116 cells, it blocks cell growth and causes cell death by gradually depleting purine levels.
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| ln Vivo |
(-)-SHIN1 has been shown to impair the growth of HCT-116 cells in culture with an IC50 of 870 nM. It is particularly effective against B cell malignancies. By inhibiting SHMT1/2, it blocks purine biosynthesis, leading to a gradual depletion of purine nucleotide pools, which in turn halts cell cycle progression and leads to cell death, specifically in cancer cells that have a high demand for one-carbon metabolism. The (-)-enantiomer is the inactive control.
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| Enzyme Assay |
A standard biochemical assay for SHMT1/2 involves measuring the conversion of [3-14C]-serine to [3-14C]-glycine. The assay is performed by incubating the recombinant enzyme (SHMT1 or SHMT2) with tetrahydrofolate (THF), pyridoxal phosphate (PLP), and varying concentrations of the test inhibitor. The reaction is initiated with the addition of [3-14C]-serine. After a set time, the product [3-14C]-glycine is separated from the substrate using a Dowex column and quantified by liquid scintillation counting to determine the IC50 value.
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| Cell Assay |
To measure the cellular activity of the compound, a metabolomics-based assay can be used. HCT-116 cells are treated with varying concentrations of (-)-SHIN1 for a defined period (e.g., 72 hours). The intracellular metabolites are then extracted and analyzed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The levels of purine nucleotides and other one-carbon pathway intermediates are quantified to assess the extent of pathway inhibition. Cell viability is measured via a standard assay like MTT to determine the half-maximal growth inhibitory concentration (GI50).
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| Animal Protocol |
The most common in vivo model for SHMT inhibitors is a xenograft model using a cancer cell line that is sensitive to SHMT inhibition, such as HCT-116, which has defects in nucleotide salvage pathways. Immunodeficient mice are implanted subcutaneously with tumor cells. Once tumors are established, the mice are treated with (-)-SHIN1 or vehicle control, typically via intraperitoneal (IP) injection or oral gavage, and tumor growth is monitored. The active enantiomer (+)-SHIN1 would be used for efficacy, while (-)-SHIN1 can serve as an inactive control.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of (-)-SHIN1 are not well described in the literature. However, it is a small molecule with a molecular weight of 400.47 daltons, suggesting potential for moderate oral absorption. The compound is formulated in vehicles like DMSO:PEG300:Tween80:ddH2O for in vivo injections. As a research chemical, its PK profile, including bioavailability, half-life, and tissue distribution, would need to be empirically determined for specific experimental designs.
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| Toxicity/Toxicokinetics |
Based on its mechanism of action as a folate-competitive inhibitor, on-target toxicity is expected to be related to the disruption of nucleotide synthesis and cell proliferation. This could affect rapidly dividing cell populations in the bone marrow, gastrointestinal tract, and immune system. The (-)-enantiomer is generally used as the inactive control in experiments, as the (+)-enantiomer is the active form. Specific toxicity data for the (-)-enantiomer is not reported, but it would be presumed to have lower toxicity than its active counterpart.
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| References | |
| Additional Infomation |
(-)-SHIN1 is the (-) enantiomer of SHIN1 (RZ-2994). The active (+) enantiomer, (+)-SHIN1, is a potent inhibitor of SHMT1/2 with IC50 values of 5 nM and 13 nM, respectively, and is used for efficacy studies. The (-)-enantiomer is the inactive form, which is structurally analogous to the active compound but has been shown to have minimal on-target activity. It serves as an essential negative control in pharmacological experiments to confirm that observed effects are due to SHMT1/2 inhibition and not off-target interactions. It is strictly a research tool, not approved for clinical use.
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| Molecular Formula |
C24H24N4O2
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|---|---|
| Molecular Weight |
400.4730
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| Exact Mass |
400.189
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| CAS # |
2444764-09-2
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| Related CAS # |
SHIN1;2146095-85-2;(+)-SHIN1;2443966-90-1
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| PubChem CID |
134337375
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| Appearance |
White to off-white solid powder
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| LogP |
4.4
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
30
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| Complexity |
707
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| Defined Atom Stereocenter Count |
1
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| SMILES |
O1C(=C(C#N)[C@@](C2=C([H])C(C([H])([H])O[H])=C([H])C(C3C([H])=C([H])C([H])=C([H])C=3[H])=C2[H])(C2=C(C([H])([H])[H])N([H])N=C12)C([H])(C([H])([H])[H])C([H])([H])[H])N([H])[H]
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| InChi Key |
VVVOFJZXKJKHTD-XMMPIXPASA-N
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| InChi Code |
InChI=1S/C24H24N4O2/c1-14(2)24(20(12-25)22(26)30-23-21(24)15(3)27-28-23)19-10-16(13-29)9-18(11-19)17-7-5-4-6-8-17/h4-11,14,29H,13,26H2,1-3H3,(H,27,28)/t24-/m1/s1
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| Chemical Name |
(4R)-6-amino-4-[3-(hydroxymethyl)-5-phenylphenyl]-3-methyl-4-propan-2-yl-2H-pyrano[2,3-c]pyrazole-5-carbonitrile
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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. |
| 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 (249.71 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.24 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.24 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (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 to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.24 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4971 mL | 12.4853 mL | 24.9707 mL | |
| 5 mM | 0.4994 mL | 2.4971 mL | 4.9941 mL | |
| 10 mM | 0.2497 mL | 1.2485 mL | 2.4971 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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