R-547 mesylate's primary targets are cyclin-dependent kinases (CDKs), including CDK1, CDK2, and CDK4. By inhibiting these kinases, it prevents the phosphorylation of their downstream targets, leading to cell cycle arrest. CDK1 is involved in the G2/M transition, CDK2 in the G1/S transition, and CDK4 in the G1 phase. Inhibition of these CDKs can inhibit tumor cell proliferation.

R547 identified as a diaminopyrimidine compound, which is a potent and selective ATP-competitive CDK inhibitor. R547 effectively inhibits CDK1/cyclinB, CDK2/cyclinE, and CDK4/cyclinD1(Ki=1-3nM) and is inactive(Ki>5,000nM) against a panel of >120 unrelated kinases. R547 effectively inhibits the proliferation of tumor cell lines independent of multidrug resistant status, histologic type, retinoblastoma protein, or p53 status, with IC50s<0.60 μM. R547 reduces phosphorylation of the cellular retinoblastoma protein at specific CDK phosphorylation sites at the same concentrations that induced cell cycle arrest, suggesting a potential pharmaco dynamics marker for clinical use. R547 inhibits the proliferation of tumor cell lines and is active in all 19 cell lines tested irrespective of tissue of origin, multidrug resistance (MDR), p53, or retinoblastoma status. R547 possessing both 5-and 6-fluoro substitution culminated in an Inhibitor with low, single-digit nanomolar potency against the CDKs(Ki=0.001,0.003,and 0.001 μM for CDK1,CDK2, and CDK4,respectively) and excellent cellular potency (IC50=0.08 μM,HCT116 cell line).
Kinase Assay: R547 is a potent ATP-competitive inhibitor of CDK1/2/4 with Ki of 2 nM/3 nM/1 nM.
Cell Assay: R547 effectively inhibits CDK1/cyclinB, CDK2/cyclinE, and CDK4/cyclinD1(Ki=1-3nM) and is inactive(Ki>5,000nM) against a panel of >120 unrelated kinases. R547 effectively inhibits the proliferation of tumor cell lines independent of multidrug resistant status, histologic type, retinoblastoma protein, or p53 status, with IC50s<0.60 μM. R547 reduces phosphorylation of the cellular retinoblastoma protein at specific CDK phosphorylation sites at the same concentrations that induced cell cycle arrest, suggesting a potential pharmaco dynamics marker for clinical use. R547 inhibits the proliferation of tumor cell lines and is active in all 19 cell lines tested irrespective of tissue of origin, multidrug resistance (MDR), p53, or retinoblastoma status. R547 possessing both 5-and 6-fluoro substitution culminated in an Inhibitor with low, single-digit nanomolar potency against the CDKs(Ki=0.001,0.003,and 0.001 μM for CDK1,CDK2, and CDK4,respectively) and excellent cellular potency (IC50=0.08 μM,HCT116 cell line).
In vitro, R-547 mesylate is a potent inhibitor of CDK1, CDK2, and CDK4. Its activity is assessed in cell-free kinase assays using purified CDK/cyclin complexes and peptide or protein substrates. It has been shown to inhibit the proliferation of various cancer cell lines by inducing cell cycle arrest.

R547 administered with oral and i.v. dosing in multiple established human tumor significantly inhibits tumor activity(P < 0.01). R547 administered orally at dose of 40 mg/kg daily in colon, lung, breast, prostate, and melanoma human tumor xenograft models shows significant TGI (79-99%). R547 is equally efficacious (TGI, 61-95%) when dosed with 40 mg/kg i.v. once weekly. These doses of R547 are not toxic and did not result in body weight loss. R547 does not show signs of overt toxicity during the course of the 3-week study and any gross pathology at necropsies done at the end of the studies. R547 inhibits tumor growth up to 95% in the HCT116 human colorectal tumor xenograft model in nude mice . R547 causes significant TGI in all of the models tested when dosed orally and i.v. at or below the maximum tolerated dose. R547 inhibits phosphorylation of retinoblastoma protein in tumors at the efficacious exposures in tumor xenograft models, providing a pharmacodynamic biomarker for clinical use. R547 reported here suggests that this is a promising molecule for evaluation in the treatment of solid tumors.

The in vitro activity of R-547 mesylate is assessed using cell-free CDK kinase assays. In these assays, purified CDK/cyclin complexes (e.g., CDK1/cyclin B, CDK2/cyclin A, CDK4/cyclin D) are incubated with a substrate (e.g., histone H1 or a peptide substrate) and ATP in the presence of varying concentrations of R-547 mesylate. The phosphorylation of the substrate is measured using a radioactive (e.g., [32P]ATP) or a fluorescence-based method. The IC50 is determined as the concentration of R-547 mesylate required to inhibit 50% of the kinase activity.

For cellular assays, various cancer cell lines (e.g., HeLa, HCT116, MCF-7) are used. Cells are cultured in appropriate media and treated with various concentrations of R-547 mesylate (typically ranging from 1 nM to 10 µM) for different time periods (e.g., 24-72 hours). Cell cycle progression is analyzed by flow cytometry using propidium iodide staining. The phosphorylation of CDK substrates (e.g., Rb protein) is assessed by Western blotting. Cell viability is assessed using the MTT or CellTiter-Glo assay.

sSspended in 1% Klucel LF in water with 0.1% Tween 80; 25 ,50,75 mg/kg; p.o.
Female nude mice bearing established HCT116 human colorectal xenografts with a mean starting volume of about 100 mm3

Specific pharmacokinetic data for R-547 mesylate is not detailed in the provided search results. As a small molecule with a molecular weight of 564.6 g/mol, it is expected to have reasonable oral bioavailability. Its half-life and metabolism would require further investigation.

Specific toxicity data for R-547 mesylate is not available in the provided search results. As a CDK inhibitor, it may have effects on rapidly dividing cells, including those in the bone marrow and gastrointestinal tract. Comprehensive toxicological studies are required to establish its full safety profile.

Mol Cancer Ther.2006 Nov;5(11):2644-58;J Med Chem.2006 Nov 2;49(22):6549-60.

R-547 mesylate is a research compound that was developed as a CDK inhibitor for the potential treatment of cancer. Its development reflects the interest in CDKs as therapeutic targets in oncology. It is not approved for clinical use.

C20H26F2N4O7S2

536.57

537.116

869369-26-6

869369-26-6 (mesylate);741713-40-6;

91618035

Typically exists as solid at room temperature

3

14

6

35

794

0

BUYYGASGVDVCPU-UHFFFAOYSA-N

InChI=1S/C18H21F2N5O4S.CH4O3S/c1-29-13-4-3-12(19)15(20)14(13)16(26)11-9-22-18(24-17(11)21)23-10-5-7-25(8-6-10)30(2,27)281-5(2,3)4/h3-4,9-10H,5-8H2,1-2H3,(H3,21,22,23,24)1H3,(H,2,3,4)

4-Piperidinamine, N-(4-amino-5-(2,3-difluoro-6-methoxybenzoyl)-2-pyrimidinyl)-1-(methylsulfonyl)-, monomethanesulfonate

R547 R 547 R-547 Ro4584820 Ro-4584820 Ro 4584820R-547 mesylate RO-4584820 mesylate.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

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

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.

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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).

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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)

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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).

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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

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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.

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 (Please use freshly prepared in vivo formulations for optimal results.)