IP; Prodrug of PGI2; Vasodilator
Beraprost targets prostacyclin (IP) receptors, which are G-protein coupled receptors (GPCRs). It also shows agonistic activity at EP3 and EP4 receptors. By binding to these receptors, Beraprost activates adenylate cyclase, increasing intracellular cAMP levels. This leads to inhibition of calcium release from intracellular storage sites and reduced calcium influx, causing relaxation of smooth muscle cells and vasodilation. The compound's dual IP/EP3 receptor affinity distinguishes it from other prostacyclin analogs.

The number of blood vessels created increased significantly after treatment with beraprost sodium (0.1, 1.0, and 10.0 μM; 24 hours), and BPS is crucial for angiogenic activity [1]. When endothelial cells were treated with beraprost sodium (0.1, 1.0, and 10.0 μM) for 24 hours, the amount of VE-cadherin in the cell-cell contact area increased, and their shape tended to be normal in contrast to cells grown in hypoxic environments [1].

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In vitro, Beraprost demonstrates potent vasodilatory effects on isolated blood vessels. The compound inhibits platelet aggregation through activation of IP receptors and elevation of cAMP levels. Beraprost also shows anti-proliferative effects on vascular smooth muscle cells. In cell-based assays, Beraprost treatment leads to increased cAMP production and downstream signaling pathway activation. The compound's activity is blocked by IP receptor antagonists, confirming receptor-mediated effects. Its in vitro potency makes it a valuable tool for studying prostacyclin signaling.

Beraprost sodium is an oral medication that is administered once day for three to seven days at a dose of 0.6 mg/kg. It has the ability to lower renal oxidative stress and further prevent renal interstitial fibrosis. fibrosis [1].

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In vivo, Beraprost exhibits potent vasodilatory and anti-platelet effects following oral administration. The compound reduces pulmonary arterial pressure in animal models of pulmonary hypertension. Beraprost also inhibits thrombosis and improves hemodynamic parameters in various cardiovascular disease models. Its oral bioavailability allows for convenient dosing in chronic studies. The compound has been evaluated in clinical trials for the treatment of pulmonary arterial hypertension and peripheral vascular disease.

Tube formation assay[1] In brief, endothelial cells (1 × 104) were seeded in a 48-well plate coated with 100 μl of growth factor-reduced Matrigel TM and incubated with and without varied concentrations of BPS (Beraprost sodium) at 0.1, 1.0, and 10.0 μmol/l for tube stabilization for 24 h at 37 °C. Tube formation was quantified by measuring the total tube loops in five random microscopic fields with a computer-assisted microscope

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The in vitro receptor binding assays for Beraprost typically involve radioligand binding studies using membrane preparations from cells expressing IP, EP3, or EP4 receptors. Competition binding assays are performed using [³H]-iloprost or other suitable radioligands to determine binding affinities (Ki values). Functional assays measure cAMP accumulation in response to Beraprost treatment. The compound is tested at concentrations ranging from 0.1 nM to 100 µM. EC₅₀ values for receptor activation are calculated from dose-response curves. Selectivity is determined by comparing potency across different receptor subtypes.

The HUVECs were cultured in a modified minimum essential medium supplemented with 10% fetal bovine serum and 1% mycillin at 37 °C in 5% CO2 and 95% air. HUVECs in hypoxia group were cultured for 12 h into an airtight humidified chamber flushed with a gas mixture containing 5% CO2, 95% N2, and 1% O2 at 37 °C. HUVECs in hypoxia + BPS group were cultured with BPS (Beraprost sodium) at 1.0 μmol/l. The cells were cultured according to the manufacturer’s instructions and the culture medium was changed every 2 or 3 days. HUVECs at passage 3 were used for the following experiments[1].

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In vitro cell-based experiments with Beraprost utilize cell lines expressing IP receptors, such as human pulmonary artery smooth muscle cells or platelets. Cells are cultured in appropriate media and treated with Beraprost at concentrations ranging from 1 nM to 10 µM. cAMP levels are measured by ELISA or radioimmunoassay. Platelet aggregation assays are performed using platelet-rich plasma. Calcium mobilization is measured using fluorescent indicators. Cell proliferation is assessed by BrdU incorporation or MTT assays. All experiments include vehicle controls and receptor antagonists to confirm specificity.

In vivo animal studies with Beraprost typically use rat or mouse models of pulmonary hypertension, thrombosis, or ischemia-reperfusion injury. Beraprost is administered orally at doses ranging from 0.1-10 mg/kg daily for 1-4 weeks. Hemodynamic parameters including blood pressure and heart rate are monitored. Platelet aggregation is measured ex vivo. Tissue samples are collected for histopathological examination and biomarker analysis. Efficacy is evaluated by reduction in pulmonary arterial pressure, prevention of thrombosis, or improvement in tissue perfusion. All procedures follow institutional animal care guidelines.

[1]. Li S, et al. Beraprost sodium mitigates renal interstitial fibrosis through repairing renal microvessels.J Mol Med (Berl). 2019 Jun;97(6):777-791.

Beraprost, a synthetic analogue of prostacyclin, is currently undergoing clinical trials for the treatment of pulmonary arterial hypertension (PAH). It is also being investigated for the prevention of reperfusion injury.

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Pharmacodynamics
Beraprost is a stable, orally effective prostacyclin analogue with vasodilatory, antiplatelet, and cytoprotective effects. Beraprost is generally well-tolerated and appears to be an effective treatment for thromboangiitis obliterans and occlusive arteriosclerosis. Comparative data from a large randomized trial showed that it was comparable to ticlopidine in treating these conditions. A randomized clinical trial reported significant efficacy of beraprost compared to placebo in patients with intermittent claudication; however, recent preliminary unpublished data from a large phase III placebo-controlled study do not support the use of beraprost in these patients. Limited data suggest that long-term treatment with beraprost may be effective for patients with pulmonary arterial hypertension (PAH), for whom treatment options are limited, and oral administration may offer a significant advantage over intravenous prostacyclin (PGI2) therapy.

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Absorption
Oral bioavailability is 50-70%.
Biological half-life
35-40 minutes
Mechanism of action
Beraprost exerts its effect by binding to prostacyclin membrane receptors, ultimately inhibiting the release of Ca2+ from intracellular storage sites. The reduction in Ca2+ influx is thought to induce smooth muscle cell relaxation and vasodilation.
Pharmacodynamics
Beraprost is a stable, orally effective prostacyclin analog with vasodilatory, antiplatelet aggregation, and cytoprotective effects. Beraprost is generally well tolerated and appears to be effective in treating patients with thromboangiitis obliterans and arteriosclerosis. Comparative data from a large randomized trial showed that its efficacy in these patients was comparable to ticlopidine. A randomized clinical trial reported significant efficacy of beraprost compared to placebo in patients with intermittent claudication; however, recent preliminary unpublished data from a large phase III placebo-controlled study do not support the use of beraprost in these patients. Limited data suggest that beraprost may be effective in the long-term treatment of patients with pulmonary hypertension. However, given the limited treatment options for patients with pulmonary hypertension, oral administration may have a significant advantage over intravenous prostacyclin (PGI2) treatment.

C24H30O5

398.499

398.209

88430-50-6

88430-50-6 (free acid);88475-69-8 (sodium);

5282428

Typically exists as solid at room temperature

1.3±0.1 g/cm3

572.1±50.0 °C at 760 mmHg

193.1±23.6 °C

0.0±1.7 mmHg at 25°C

1.625

2.87

3

5

8

29

659

5

CC#CCC(C)C(/C=C\C1C(CC2C1C3=CC=CC(=C3O2)CCCC(=O)O)O)O

CTPOHARTNNSRSR-OUKQBFOZSA-N

InChI=1S/C24H30O5/c1-3-4-7-15(2)19(25)13-12-17-20(26)14-21-23(17)18-10-5-8-16(24(18)29-21)9-6-11-22(27)28/h5,8,10,12-13,15,17,19-21,23,25-26H,6-7,9,11,14H2,1-2H3,(H,27,28)/b13-12+

4-[2-hydroxy-1-[(E)-3-hydroxy-4-methyloct-1-en-6-ynyl]-2,3,3a,8b-tetrahydro-1H-cyclopenta[b][1]benzofuran-5-yl]butanoic acid

ML-1229; MDL-201229; beraprost; 88430-50-6; 88430-50-6 (free acid); 4-[2-hydroxy-1-[(E)-3-hydroxy-4-methyloct-1-en-6-ynyl]-2,3,3a,8b-tetrahydro-1H-cyclopenta[b][1]benzofuran-5-yl]butanoic acid; Beraprost (USAN); 1H-Cyclopenta(b)benzofuran-5-butanoic acid, 2,3,3a,8b-tetrahydro-2-hydroxy-1-(3-hydroxy-4-methyl-1-octen-6-ynyl)-; 4-(2-Hydroxy-1-(3-hydroxy-4-methyloct-1-en-6-yn-1-yl)-2,3,3a,8b-tetrahydro-1H-cyclopenta[b]benzofuran-5-yl)butanoic acid; 4-(1,2,3a,8b-tetrahydro-2-hydroxy-1-(3-hydroxy-4-methyloct-6-yne-1-enyl)-5-cyclopenta(b)benzofuranyl)butyrate; MDL 201229

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.)