In LPS-stimulated RAW264.7 macrophages, Fimasartan (62.5, 125, or 250 μM) potently and concentration-dependently inhibited LPS-induced nitric oxide (NO) production. The positive control NIL (20 μM) also inhibited NO production [4].
Fimasartan (62.5, 125, or 250 μM) concentration-dependently inhibited the LPS-induced up-regulation of iNOS protein expression in RAW264.7 macrophages as determined by Western blot analysis [4].
Fimasartan (62.5, 125, or 250 μM) significantly reduced LPS-induced iNOS mRNA levels in RAW264.7 macrophages as determined by qRT-PCR [4].
Fimasartan (62.5, 125, or 250 μM) concentration-dependently inhibited the LPS-induced increase in iNOS promoter activity in RAW264.7 macrophages using a luciferase reporter gene assay [4].
Fimasartan (62.5, 125, or 250 μM) prevented LPS-induced increases in NF-κB-dependent luciferase activity in RAW264.7 macrophages [4].
Fimasartan (250 μM) significantly reduced LPS-induced NF-κB-DNA binding activity at 30 or 60 minutes as measured by EMSA in RAW264.7 macrophages [4].
Fimasartan (62.5, 125, or 250 μM) concentration-dependently reduced LPS-induced NF-κB-DNA binding activity at 60 minutes and significantly suppressed LPS-induced nuclear translocation of the p65 and p50 subunits of NF-κB [4].
Fimasartan (62.5, 125, or 250 μM) significantly decreased LPS-induced transcription activity of AP-1 [4].
Fimasartan (250 μM) reduced LPS-induced AP-1-DNA binding activity at 30 or 60 minutes as measured by EMSA in RAW264.7 macrophages [4].
Fimasartan (62.5, 125, or 250 μM) concentration-dependently attenuated LPS-induced nuclear translocation of c-Fos and c-Jun (AP-1 subunits) [4].
In vitro transport studies using Xenopus laevis oocytes expressing OATP1B1 showed that Fimasartan inhibited the uptake of the OATP1B1 substrate estrone-3-sulfate (ES). The IC50 value for this inhibition was 11.6 ± 2.4 μM [3].

Pretreatment with Fimasartan increased the survival rates of mice with established LPS-induced endotoxaemia in a murine model of sepsis [4].

To evaluate inhibition of OATP1B1-mediated uptake, cRNA encoding the OATP1B1 transporter was synthesized and injected into Xenopus laevis oocytes, which were incubated for 2 days at 18°C to ensure high expression. The specific uptake of a known OATP1B1 substrate, [3H]estrone-3-sulfate (ES), was quantified in control oocytes. To assess inhibition, different concentrations of Fimasartan (0.1 - 200 μM) were added along with 35 nM [3H]ES to the oocytes. After 30-minute incubations, the reactions were terminated by adding ice-cold ND96 solution. Oocytes were washed 5 times, then individual oocytes were crushed and the associated radioactivity was quantified in a liquid scintillation analyzer. Initial ES uptake rates (ν) were determined for the various concentrations of Fimasartan and expressed as a percentage of the control uptake (in the absence of Fimasartan). [3].

RAW264.7 macrophages were plated and incubated with or without LPS (1 μg/mL) in the absence or presence of various concentrations (62.5, 125, or 250 μM) of Fimasartan for 24 h. The nitrite accumulated in the culture medium was measured based on the Griess reaction. An equal volume of cell culture medium was mixed with Griess reagent, incubated at room temperature for 10 min, and the absorbance at 540 nm was measured [4].
Cells were pretreated with Fimasartan (62.5, 125, or 250 μM) for 1 h and then treated with LPS (1 μg/mL) for 24 h. Total cellular proteins were extracted, resolved by SDS-PAGE, and electroblotted onto a PVDF membrane. The immunoblot was incubated with primary antibodies for iNOS, p65, c-Jun, c-Fos, PARP, or β-actin, followed by incubation with a horseradish peroxidase-conjugated secondary antibody. Blots were developed by enhanced chemiluminescence [4].
Total cellular RNA was isolated from cells stimulated with LPS (1 μg/mL) in the presence or absence of Fimasartan (62.5, 125, or 250 μM) for 4 h. RNA was reverse-transcribed using MuLV reverse transcriptase, and PCR amplification was performed using SYBR green incorporation with specific primers for iNOS and β-actin. Steady-state mRNA levels were determined by quantitative real-time PCR [4].
For the iNOS promoter activity assay, RAW264.7 macrophages were co-transfected with a pGL3-iNOS promoter (-1592/+185) reporter plasmid vector and the phRL-TK plasmid as an internal control. After transfection, cells were pretreated with Fimasartan for 1 h and then stimulated with LPS (1 μg/mL) for 18 h. Cells were lysed and luciferase activity was determined using a luciferase assay system [4].
For the NF-κB and AP-1 reporter gene assays, cells were transiently co-transfected with a pNF-κB-luc or pAP-1-luc reporter plasmid vector plus the phRL-TK plasmid. After transfection, cells were pretreated with various concentrations (62.5, 125, or 250 μM) of Fimasartan for 1 h and then stimulated with LPS (1 μg/mL) for 18 h. Cells were harvested and luciferase activities were determined [4].
For EMSA, nuclear extracts were prepared from cells treated with Fimasartan (62.5, 125, or 250 μM) for 1 h and then stimulated with LPS (1 μg/mL) for 30 or 60 min. Nuclear extracts (5 μg) were mixed with double-stranded biotin end-labeled NF-κB or AP-1 oligonucleotides. DNA-binding activities were measured using a chemiluminescent EMSA kit [4].

The study used a murine model of LPS-induced sepsis. No details on drug formulation, dose, route, or frequency of Fimasartan administration were provided in the available text [4].
The human study for pharmacokinetics was an open-label, single-dose, parallel study. After an overnight fast of 10 h, subjects received a single 240-mg oral dose of Fimasartan taken with 240 ml of water. Food intake was permitted 4 h after dosing [1].
The drug interaction study was an open-label, 2-period, 2-sequence, crossover, multiple-dosing study. In one period, a group of subjects received a single daily oral dose of 80 mg atorvastatin for 7 days, and another group received 80 mg atorvastatin with 240 mg Fimasartan for 7 days. The first period was followed by a 1-week washout period, after which volunteers were administered the opposite treatments. On the seventh day of each period, subjects were dosed after overnight fasting, which lasted until 4 hours after administration [3].

Absorption, Distribution and Excretion
Time to Peak (Tmax) is 0.5–1.3 hours. Most is excreted unchanged via bile, with less than 3% excreted in urine. Biological Half-Life Elimination half-life is 7–10 hours.

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After oral administration of a single 240 mg dose of Fimasartan to healthy young (19-45 years) and older (≥65 years) male subjects: In older subjects (n=10), the mean AUC0-∞ was 2899.0 ng/mL/h, which was significantly greater than in young subjects (n=12) where it was 1767.4 ng/mL/h (p = 0.03). The geometric mean AUC0-∞ was 69.4% higher in older subjects. The geometric mean Cmax was 81.2% higher in older subjects (1103.5 ± 951.7 ng/mL) than in young subjects (658.2 ± 684.5 ng/mL), but this difference was not statistically significant (p = 0.107). Median tmax was 1.0 hour (range 0.5-6.0) for older and 1.0 hour (range 0.5-3.0) for young subjects. Mean elimination half-life (t1/2) was 6.6 ± 1.3 h in older and 6.0 ± 1.7 h in young subjects (p = 0.203). Mean apparent oral clearance (CL/F) was 1688.4 ± 1074.0 mL/min in older and 2797.7 ± 1201.0 mL/min in young subjects (p = 0.03). Mean apparent volume of distribution (Vd/F) was 973.7 ± 705.2 L in older and 1481.8 ± 882.9 L in young subjects (p = 0.069) [1].
In a previous study cited in the literature, after oral administration of Fimasartan to rats and dogs, systemic bioavailability ranged from 34.2 to 37.7%, and most of the radioactive dose (97.3%) was excreted in feces (72 h post-dose) via bile in rats, with unchanged parent drug being the major circulating component in the plasma [1].
According to the investigator's brochure and a report by Lane et al., peak plasma concentrations of Fimasartan have been observed between 0.5 and 3 h after dosing. Fimasartan is highly bound to plasma proteins (95.6 - 97.2%). After a single oral administration, it exhibits dose-dependent pharmacokinetics over the 20- to 480-mg dose range, and the mean elimination half-life is about 9-16 h. After repeated dosing, steady-state plasma concentrations are reached in 3 days, with no accumulation observed [1].
Fimasartan is metabolized primarily by cytochrome P450 3A (CYP3A). It acts both as a substrate and as an inhibitor of organic anion-transporting polypeptide 1B1 (OATP1B1; SLC21A6). Less than 3% of the Fimasartan dose is recovered in the urine over the first 24 hours post-dose [3].
Coadministration of Fimasartan (240 mg) with atorvastatin (80 mg) increased the atorvastatin acid geometric mean Cmax,ss by 1.89-fold (95% CI: 1.49-2.39) and AUCτ,ss by 1.19-fold (0.96-1.48) compared with atorvastatin alone. Fimasartan also increased the mean 2-hydroxy atorvastatin acid Cmax,ss and AUCτ,ss by 2.45-fold (1.80-3.35) and 1.42-fold (1.09-1.85), respectively [3].

In the single-dose study (240 mg), Fimasartan was well tolerated. Of 22 subjects, 8 (36.4%) experienced 13 treatment-emergent adverse events (TEAEs). Four events (1 dizziness, 1 headache, and 2 CPK elevations) occurred in the young group, were considered probably related to medication, and were mild-to-moderate in severity. No TEAE was serious or severe, except one older subject with a pre-existing left inguinal hernia. Reductions in blood pressure were mild, transient, and asymptomatic in most subjects [1].
In the drug interaction study (multiple doses of 80 mg atorvastatin with or without 240 mg Fimasartan), a total of 39 adverse events (AEs) were reported in 17 of 30 subjects. Most AEs were transient, and all were mild in intensity, with no serious AEs. Three subjects discontinued due to AEs (one urticaria, two with increased blood CPK levels and myalgia, likely from strenuous exercise). There were no significant changes in vital signs, ECGs, or laboratory tests [3].
Fimasartan exhibits >95% protein binding capacity [3].

[1]. Fimasartan, anti-hypertension drug, suppressed inducible nitric oxide synthase expressions via nuclear factor-kappa B and activator protein-1 inactivation. Biol Pharm Bull. 2013;36(3):467-74.

[2]. Pharmacological characterization of BR-A-657, a highly potent nonpeptide angiotensin II receptor antagonist. Biol Pharm Bull. 2013;36(7):1208-15.

[3]. Effect of age on the pharmacokinetics of fimasartan (BR-A-657). Expert Opin Drug Metab Toxicol. 2011 Nov;7(11):1337-44.

[4]. The effect of the newly developed angiotensin receptor II antagonist fimasartan on the pharmacokinetics of atorvastatin in relation to OATP1B1 in healthy male volunteers. J Cardiovasc Pharmacol. 2011 Nov;58(5):492-9.

Finmasartan belongs to the biphenyl class of drugs. Finmasartan is an angiotensin II receptor antagonist (ARB) used to treat hypertension and heart failure. Clinical trials have shown that the combination of fimasartan and hydrochlorothiazide (a diuretic) is safe and effective. Finmasartan was approved for marketing in South Korea on September 9, 2010, and is marketed by Boling Pharmaceutical under the brand name Kanarb. Indications: For the treatment of hypertension and heart failure. Mechanism of Action: Angiotensin II activates AR1 receptors, leading to vasoconstriction and increased norepinephrine release. Norepinephrine further enhances vasoconstriction by acting on α1-adrenergic receptors. It also stimulates aldosterone secretion, thereby increasing the reabsorption of sodium and water by the renal tubules. Finmasartan binds to and antagonizes angiotensin II receptor 1 (AR1), thereby preventing vasoconstriction and reducing aldosterone secretion, increasing sodium excretion, and ultimately leading to a decrease in blood volume. These effects work together to produce a blood pressure-lowering effect.

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Pharmacodynamics
Fimasartan is a selective angiotensin receptor 1 (AR1) inhibitor. It lowers blood pressure by inhibiting vasoconstriction.

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Fimasartan is approved by the Korean Food and Drug Administration (KFDA) for the treatment of essential hypertension [1][4].
Inflammatory processes are important in hypertension pathophysiology. Angiotensin II is a pro-inflammatory factor, and ARBs like Fimasartan may exert general anti-inflammatory effects beyond blood pressure control. Fimasartan suppresses iNOS expression via NF-κB and AP-1 inactivation in macrophages, suggesting potential to ameliorate inflammatory disease [4].
The effect of age on the PK of Fimasartan was modest (AUC increase < twofold in older subjects) [1].
The interaction between Fimasartan and atorvastatin is considered weak (AUC increase < 2.0-fold), but careful patient assessment is recommended when coadministering [3].
Fimasartan exhibits little capacity to inhibit or induce cytochrome P450 enzymes [3].

C27H31N7OS

501.65

501.231

247257-48-3

Fimasartan-d6

9870652

Light yellow to yellow solid powder

1.3±0.1 g/cm3

693.0±65.0 °C at 760 mmHg

372.9±34.3 °C

0.0±2.2 mmHg at 25°C

1.658

4.53

1

6

9

36

849

0

AMEROGPZOLAFBN-UHFFFAOYSA-N

InChI=1S/C27H31N7OS/c1-5-6-11-24-28-18(2)23(16-25(36)33(3)4)27(35)34(24)17-19-12-14-20(15-13-19)21-9-7-8-10-22(21)26-29-31-32-30-26/h7-10,12-15H,5-6,11,16-17H2,1-4H3,(H,29,30,31,32)

2-(2-Butyl-4-methyl-6-oxo-1-{[2'-(1H-tetrazol-5-yl)-4-biphenylyl]methyl}-1,6-dihydro-5-pyrimidinyl)-N,N-dimethylethanethioamide

BR-A657; BR A657; BRA 657; BR-A-657.K; BRA657; Fimasartan. brand name: Kanarb.

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)

DMSO : ≥ 49 mg/mL (~97.68 mM)

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.)