| Size | Price | Stock | Qty |
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| 5mg |
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| Targets |
ADAMTS-5 inhibitors: ADAMTS-5 (aggrecanase-2) [1]
ADAMTS-4: Compound 12 showed functional selectivity over ADAMTS-4 (aggrecanase-1) with an IC50 of 44 µM [1] ADAMTS-4: Compound 14 showed an IC50 of 7.5 µM against ADAMTS-4 [1] |
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| ln Vitro |
ADAMTS-5 (protein aggregase-2) is an external matrix-degrading protease that causes cartilage degradation in arthritis [2].
ADAMTS-5 Enzyme Inhibition: The compounds were evaluated for their ability to inhibit ADAMTS-5. Compound 12 exhibited potent inhibitory activity with an IC50 of 1.1 µM. Other active compounds included 14 (IC50: 2.2 µM), 17 (IC50: 2.5 µM), 18 (IC50: 4.2 µM), 20 (IC50: 2.1 µM), and 21 (IC50: 3.7 µM) [1]. ADAMTS-4 Enzyme Inhibition: Compound 12 showed an IC50 of 44 µM against ADAMTS-4, demonstrating >40-fold functional selectivity over ADAMTS-5. Compound 14 showed an IC50 of 7.5 µM against ADAMTS-4, demonstrating lower selectivity [1]. Structure-Activity Relationship (SAR): The presence of a para-chloro (p-Cl) moiety on the benzyl group (as in compound 12) was crucial for potent ADAMTS-5 inhibition. Moving the chloro substituent to the ortho-position (compound 13) resulted in a ~20-fold loss in activity (IC50: 23.2 µM). Replacing the benzyl methylene with a direct phenyl link (compound 16) reduced activity. The 3-CF3 moiety on the pyrazole could be replaced with other substituents like phenyl to maintain activity (e.g., compound 21) [1]. Binding Mechanism: Compounds 12 and 14 successfully displaced a xanthene-ADAMTS-5 hydroxamic acid inhibitor from the ADAMTS-5 protein, indicating that their inhibitory activity involved direct binding to ADAMTS-5 [1]. |
| ln Vivo |
The study references previous findings that Adamts5 knockout mice are protected from cartilage destruction in arthritis models and that knock-in of an aggrecan cleavage site mutation is protective. [2]
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| Enzyme Assay |
ADAMTS-5 FRET Assay: ADAMTS-5 activity was determined using a fluorescence resonance energy transfer (FRET) assay. The assay utilized a QF-peptide that contained the aggrecanase cleavage site. This method was used to measure the functional inhibition of ADAMTS-5 by the compounds. The results were reported as IC50 values [1].
ADAMTS-4 FRET Assay: Several compounds were analyzed for ADAMTS-4 activity using the same QF-peptide substrate that was used for the ADAMTS-5 activity studies. This allowed for a direct comparison of inhibitory potency and selectivity between the two enzymes [1]. Fluorescence Polarization Binding Assay: Binding to target studies were conducted using fluorescence polarization experiments. These experiments employed a xanthene-ADAMTS-5 hydroxamic acid inhibitor as a probe to assess whether the thioxothiazolidin-4-one compounds directly bound to the ADAMTS-5 protein [1]. |
| Animal Protocol |
Ex Vivo Joint Inflammation Model: A novel ex vivo knee inflammation model was developed. Lower limbs from 8-week-old male Adamts5+/- mice were dissected. After mid-diaphyseal tibial and femoral osteotomy, the knee joint was sectioned sagittally in the midline. The explants were washed with serum-free medium and then cultured in DMEM containing 10% fetal bovine serum for 48 hours at 37°C in 5% CO2. After 48 hours, the medium was replaced with serum-free DMEM containing antibiotics, with or without 10 ng/ml of IL-1α and 20 ng/ml of TNFα. Explants were cultured for 16, 48, or 96 hours. For each mouse, one hind limb was treated with cytokines, while the contralateral joint was cultured in serum-free medium as a control. [2]
Ex Vivo Joint Inflammation Model: A novel ex vivo knee inflammation model was developed. Lower limbs from 8-week-old male Adamts5+/- mice were dissected. After mid-diaphyseal tibial and femoral osteotomy, the knee joint was sectioned sagittally in the midline. The explants were washed with serum-free medium and then cultured in DMEM containing 10% fetal bovine serum for 48 hours at 37°C in 5% CO2. After 48 hours, the medium was replaced with serum-free DMEM containing antibiotics, with or without 10 ng/ml of IL-1α and 20 ng/ml of TNFα. Explants were cultured for 16, 48, or 96 hours. For each mouse, one hind limb was treated with cytokines, while the contralateral joint was cultured in serum-free medium as a control. [2] |
| References |
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| Additional Infomation |
Background and Disease Context: Osteoarthritis (OA) is a disease characterized by the degradation of aggrecan and collagen in articular cartilage. Aggrecan loss occurs in the initial phases of the disease. Inhibiting aggrecan breakdown could represent a disease-modifying osteoarthritis treatment, in contrast to therapies that only provide symptomatic relief [1].
Target Rationale: Aggrecan is catabolized by ADAMTS-4 (aggrecanase-1) and ADAMTS-5 (aggrecanase-2). While both enzymes cleave aggrecan at the Glu373-Ala374 bond, studies in knockout mice suggest ADAMTS-5 could be the primary aggrecanase in cartilage, as ADAMTS-5 knockout mice showed reduced OA severity, whereas ADAMTS-4 knockouts showed normal progression [1]. Compound Significance: To the authors' knowledge, these compounds represent the first disclosure of inhibitors of ADAMTS-5 [1]. |
| Molecular Formula |
C16H11CLF3N3OS3
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|---|---|
| Molecular Weight |
449.91
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| Exact Mass |
448.97
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| CAS # |
929634-33-3
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| PubChem CID |
16664963
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.59±0.1 g/cm3 (20 °C, 760 mmHg)
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| LogP |
5.202
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
27
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| Complexity |
626
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1C(=C(C(=N1)C(F)(F)F)/C=C\2/C(=O)NC(=S)S2)SCC3=CC=C(C=C3)Cl
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| InChi Key |
CVYPRDPBCXSVBN-WDZFZDKYSA-N
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| InChi Code |
InChI=1S/C16H11ClF3N3OS3/c1-23-14(26-7-8-2-4-9(17)5-3-8)10(12(22-23)16(18,19)20)6-11-13(24)21-15(25)27-11/h2-6H,7H2,1H3,(H,21,24,25)/b11-6-
|
| Chemical Name |
(5Z)-5-[[5-[(4-chlorophenyl)methylsulfanyl]-1-methyl-3-(trifluoromethyl)pyrazol-4-yl]methylidene]-2-sulfanylidene-1,3-thiazolidin-4-one
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| Synonyms |
ADAMTS-5 Inhibitor ADAMTS5 Inhibitor ADAMTS 5 Inhibitor
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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 |
| 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 : ~83.33 mg/mL (~185.21 mM)
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2227 mL | 11.1133 mL | 22.2267 mL | |
| 5 mM | 0.4445 mL | 2.2227 mL | 4.4453 mL | |
| 10 mM | 0.2223 mL | 1.1113 mL | 2.2227 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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