| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
OCI-101 does not have a defined direct molecular target in this study. It inhibits osteoclast differentiation by modulating multiple signaling pathways downstream of RANKL and M-CSF. Specifically, it inhibits the phosphorylation of IκBα, p65, and ERK in the RANKL-induced pathway, and inhibits the phosphorylation of JNK, Akt, and ERK in the M-CSF-induced pathway. The study does not report IC50, Ki, or EC50 values for these effects. [1]
The primary biological targets of 1,3-dibenzyl-5-fluorouracil are the receptor activator of NF-κB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) signaling pathways, which are critical for osteoclast differentiation and function. Unlike 5-fluorouracil which exerts its effects as a thymidylate synthase inhibitor, this derivative acts as a chemical suppressor of osteoclastogenesis by inhibiting the expression of osteoclast markers and reducing the formation of multinucleated osteoclasts. The compound targets the cellular processes involved in bone resorption rather than nucleic acid synthesis, representing a distinct mechanism from its parent compound. |
|---|---|
| ln Vitro |
- Inhibition of Osteoclast Differentiation: OCI-101 treatment (0-100 μM) dose-dependently reduced the formation of multinucleated, TRAP-positive osteoclasts from bone marrow macrophages (BMMs) stimulated with M-CSF (50 ng/ml) and RANKL (200 ng/ml) for 4 days. The number of TRAP+ multinucleated cells (≥3 nuclei) and total TRAP activity were significantly decreased. [1]
- Inhibition of Bone Resorption: OCI-101 significantly reduced the resorption pit area formed by mature osteoclasts on a bone substrate, as quantified by ImageJ software. [1] - Inhibition of Osteoclast Marker Gene Expression: Real-time PCR analysis showed that OCI-101 inhibited the expression of osteoclast-specific marker genes (e.g., TRAP, cathepsin K) in a dose-dependent manner. [1] - Effect on RANKL-Induced Signaling: In BMMs pre-incubated with 100 μM OCI-101 for 12 hours, subsequent RANKL stimulation (200 ng/ml) resulted in decreased phosphorylation of IκBα, p65, and ERK compared to control. Phosphorylation of JNK was unaffected, while p38 phosphorylation was slightly enhanced. [1] - Effect on M-CSF-Induced Signaling: Under the same pre-incubation conditions, M-CSF stimulation (50 ng/ml) led to decreased phosphorylation of JNK, Akt, and ERK in OCI-101-treated cells. Phosphorylation of IκBα, p65, and p38 was unaffected. [1] - Cytotoxicity: OCI-101 showed no significant cytotoxicity in BMMs at concentrations up to 100 μM after 4 days of culture, as measured by a Cell Counting Kit-8 assay. [1] |
| ln Vivo |
- Inhibition of OVX-Induced Bone Loss in Mice: In an ovariectomy (OVX)-induced osteoporosis model, daily intraperitoneal injection of OCI-101 (10 mg/kg, but not 1 mg/kg) for 3 weeks significantly prevented trabecular bone loss in the femur. Histological analysis showed that the number of TRAP+ osteoclasts in the trabecular region was decreased by 26.9% (p<0.05) in the OCI-101 (10 mg/kg)-treated group compared to the OVX control group. [1]
- Micro-CT Analysis: Micro-computed tomography (micro-CT) analysis confirmed the protective effect. In OVX mice treated with OCI-101 (10 mg/kg), bone mineral density (BMD) was 12.8% higher (p<0.001), trabecular bone volume fraction (BV/TV) was 34.8% higher (p<0.01), trabecular number (Tb. N) was 32.9% higher (p<0.01), and trabecular spacing (Tb. Sp) was 17.8% lower (p<0.05) compared to vehicle-treated OVX controls. [1] |
| Enzyme Assay |
Specific non-cellular enzyme/receptor binding assays for 1,3-dibenzyl-5-fluorouracil are not detailed in the available literature. However, standard protocols for assessing osteoclastogenesis inhibition typically involve evaluating the RANKL-RANK interaction using cell-free systems. A representative approach includes using purified RANKL protein immobilized on assay plates, incubating with varying concentrations of the test compound, and detecting binding using labeled anti-RANKL antibodies. Alternatively, NF-κB activation assays in cell-free nuclear extracts can be performed, where the compound's ability to inhibit RANKL-induced p65 nuclear translocation is measured by ELISA-based transcription factor assays. These methods allow quantification of the compound's interference with the initial signaling events in osteoclast differentiation.
|
| Cell Assay |
- Osteoclast Differentiation Assay: Bone marrow cells from 6-week-old male C57BL/6J mice were cultured at 1x10⁵ cells/well in 96-well plates. They were differentiated into bone marrow-derived osteoclasts (BMOCs) using 50 ng/ml M-CSF and 200 ng/ml RANKL for 4 days, in the presence or absence of OCI-101 (0-100 μM). Cells were then fixed and subjected to TRAP staining (using naphthol AS phosphate and fast red violet) and TRAP solution assay (using p-nitrophenyl phosphate substrate, with absorbance read at 405 nm). TRAP-positive multinucleated cells (TRAP-MNCs) with >3 nuclei were counted as mature osteoclasts. [1]
- Pit Formation Assay (Bone Resorption): Mature osteoclasts were cultured on bone substrate with or without OCI-101. The resorbed pits were visualized, and the resorption area was quantified using ImageJ software. [1] - Real-Time PCR for OC Markers: Total RNA was isolated from differentiated BMOCs, and real-time PCR was performed to measure the expression of osteoclast-specific genes (e.g., TRAP, cathepsin K). Expression levels were normalized to β-actin. [1] - Western Blot (Signaling Pathway Analysis): BMMs were pre-incubated with 100 μM OCI-101 for 12 hours. After a 2-hour starvation period without RANKL or M-CSF, cells were stimulated with RANKL (200 ng/ml) or M-CSF (50 ng/ml) for the indicated times. Cell lysates were subjected to SDS-PAGE and immunoblotting with antibodies specific for phosphorylated and total forms of IκBα, p65, p38, ERK, JNK, and Akt. β-Actin served as the loading control. [1] - Cytotoxicity Assay: BMMs were cultured with M-CSF alone (50 ng/ml) and various concentrations of OCI-101 (0-100 μM) for 4 days. Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) according to the manufacturer's protocol. [1] |
| Animal Protocol |
- Animals: 8-week-old female C57BL/6J mice (n=10-11 per group) were used. [1]
- Ovariectomy (OVX) Model: Mice were randomly divided into four groups: Sham operation (Sham), OVX with vehicle control, OVX + low-dose OCI-101 (1 mg/kg), and OVX + high-dose OCI-101 (10 mg/kg). OVX was performed by surgical removal of ovaries, while sham surgery involved identifying both ovaries without removal. [1] - Drug Administration: One week after surgery, mice received daily intraperitoneal (i.p.) injections of OCI-101 (1 or 10 mg/kg) or vehicle for 3 weeks. [1] - Tissue Collection and Analysis: After the treatment period, femurs were collected and fixed with 10% formalin. For micro-CT analysis, the trabecular morphometry of the distal femur was analyzed using a SkyScan 1076 micro-CT scanner. For histological analysis, femurs were decalcified in 15% EDTA, embedded in paraffin, and sectioned. Sections were stained with TRAP and hematoxylin, and TRAP-positive osteoclasts were counted under a microscope. [1] |
| ADME/Pharmacokinetics |
Specific pharmacokinetic parameters for 1,3-dibenzyl-5-fluorouracil have not been fully characterized in publicly available literature. However, based on its physicochemical properties, the compound has a calculated LogP value of 2.245, indicating moderate lipophilicity that may facilitate cellular membrane permeation. The molecular weight of 310.32 g/mol falls within the typical range for orally bioavailable small molecules. For research applications, storage recommendations include powder stability at -20°C for up to 3 years and solutions at -80°C for up to 6 months. The compound shows solubility in chloroform, methanol, and other organic solvents, with limited water solubility as suggested by its structure featuring two benzyl groups.
|
| Toxicity/Toxicokinetics |
- In Vitro Cytotoxicity: No significant cytotoxicity was observed in bone marrow macrophages (BMMs) treated with OCI-101 at concentrations up to 100 μM for 4 days, as determined by a CCK-8 assay. [1]
- In Vivo Tolerability: The document states that OCI-101 was injected intraperitoneally at doses of 1 and 10 mg/kg daily for 3 weeks. No overt signs of toxicity, such as significant body weight loss or mortality, were reported. The study was approved by the Animal Experiment Ethics Committee of Chungnam National University. [1] |
| References | |
| Additional Infomation |
- Mechanism of Action: OCI-101 inhibits osteoclastogenesis by downregulating multiple signaling pathways downstream of both RANKL and M-CSF. Specifically, it blocks RANKL-induced phosphorylation of ERK, IκBα, and p65, and M-CSF-induced phosphorylation of ERK, JNK, and Akt. This leads to decreased expression of key osteoclastogenic transcription factors and OC-specific genes. [1]
- Therapeutic Potential: Given its efficacy in preventing OVX-induced bone loss in mice, OCI-101 is proposed as a good drug candidate for treating postmenopausal osteoporosis and other inflammatory bone-related diseases characterized by excessive osteoclast activity. [1] - Discovery Method: OCI-101 was identified by screening a chemical library of 16,380 unique compounds from the Korea Chemical Bank (KCB) using an osteoclast differentiation assay. [1] |
| Molecular Formula |
C18H15FN2O2
|
|---|---|
| Molecular Weight |
310.32
|
| Exact Mass |
310.112
|
| CAS # |
75500-02-6
|
| PubChem CID |
348853
|
| Appearance |
Typically exists as solid at room temperature
|
| Density |
1.32g/cm3
|
| Boiling Point |
441.1ºC at 760 mmHg
|
| Flash Point |
220.6ºC
|
| Index of Refraction |
1.643
|
| LogP |
2.245
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
23
|
| Complexity |
479
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1=CC=C(C=C1)CN2C=C(C(=O)N(C2=O)CC3=CC=CC=C3)F
|
| InChi Key |
QVRHSLFHSJNNKP-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C18H15FN2O2/c19-16-13-20(11-14-7-3-1-4-8-14)18(23)21(17(16)22)12-15-9-5-2-6-10-15/h1-10,13H,11-12H2
|
| Chemical Name |
1,3-dibenzyl-5-fluoropyrimidine-2,4-dione
|
| Synonyms |
1,3-DIBENZYL-5-FLUOROURACIL; 75500-02-6; DTXSID00325048; RefChem:218359; DTXCID80276165;
|
| 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)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| 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 | 3.2225 mL | 16.1124 mL | 32.2248 mL | |
| 5 mM | 0.6445 mL | 3.2225 mL | 6.4450 mL | |
| 10 mM | 0.3222 mL | 1.6112 mL | 3.2225 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
