Osthole

Histamine H₁ receptor

Osthol is an O-methylated coumarin present in plants, including Angelica pubescens, Cnidium monnieri, and Angelica archangelica. In rabbit platelets that have been cleaned, osthol prevents platelet aggregation and ATP release that is brought on by ADP, arachidonic acid, PAF, collagen, ionophore A23187, and thrombin. Through the extracellular signal-regulated kinase 1/2 and bone morphogenetic protein-2/p38 pathways, osteospot mediates cell differentiation in human osteoblast cells and holds promise as a treatment for osteoporosis. By making HBsAg more glycosylated, osteoporosis also inhibits the hepatitis B virus's secretion in cell culture. [3]

Local bone formation was significantly stimulated by subcutaneous injection of osthole at a dose of 5 mg/kg per day into mouse skulls (histological analysis of skull tissue samples collected 2 weeks after the last injection and stained with H&E Orange G). Osthole significantly influenced bone formation, according to morphological analysis, and microtubule inhibition of TN-16 was just as successful as it was in the earlier investigation. However, when osthole was taken daily at a dose of 1 mg/kg, no such effect was observed. The bone loss of the castration stent can be considerably reversible with an 8-week intraperitoneal injection of osthole. Histological analysis of L4 samples stained with trinitrophenol poinsettia revealed that castrate stents treated with osthole had partially recovered their trabecular structure. Osthole treatment dramatically increased trabecular volume, thickness, and total BMD while decreasing trabecular separation, according to morphological analysis [2].

On the first study day, participants' peripheral blood samples are drawn between 7 and 9 a.m. and transferred into grouping tubes containing K₃EDTA. And then they make fresh PBMCs. A solution of 1% heat-inactivated human AB serum, 1% gentamicin, and 0.25% PHA is added to isolated cells that are seeded on 24-well plates at a density of 1×10⁶ per well using RPMI-1640. A 24-hour period is given to each well before active reagents are added, with pure medium serving as the substance's control. After three additional days, cells are harvested[1].

Mice: Four-week-old ICR Swiss mice receive subcutaneous injections over the calvarial surface twice a day for five days straight, either with or without Osthole treatment. The doses are 1 and 5 mg/kg per day, with three mice per group. As a positive control, microtubule inhibitor TN-16 (5 mg/kg per day, subcutaneous injection, twice daily for 2 days; 3 mice per group) is used. Three weeks after the start of treatment, all the mice are put to sleep, and the calvariae are removed, preserved for two days in 10% phosphate-buffered formalin, decalcified for two weeks in 10% EDTA, and then embedded in paraffin. Hematoxylin and eosine orange G are used to cut and stain histologic sections. Using the OsteoMeasure System for histomorphometry, the amount of new bone over the calvarial surface is measured. In order to quantify the mineral appositional rate (MAR) and bone-formation rate (BFR), mice undergo intraperitoneal injections of 20 mg/kg of double calcein at days 7 and 14, following which they are put to death 7 days later. Plastic sections of the labeling are inspected. The calvarial samples that have been dissected are embedded in methyl methacrylate after being fixed in 75% ethanol. A fluorescent microscope is used to examine unstained transverse sections that have a thickness of 3 µm. With the OsteoMeasure System, MAR and BFR are measured.
Rats: The rats are thirty six-month-old female Sprague-Dawley rats. The rats are randomly assigned by body weight into three groups for the surgery (n=10): group 1 is a sham surgery followed by PBS vehicle treatment (sham+VEH); group 2 is an ovariectomy followed by vehicle treatment (OVX+VEH); and group 3 is an ovariectomy followed by Osthole treatment (OVX+OST). The rats are given an intraperitoneal nembutal injection (30 mg/kg) to induce anesthesia. The eight-week course of treatment is administered beginning one month following surgery. For eight weeks, either vehicle or Osthole (100 mg/kg daily) is taken orally once daily. Dual-energy X-ray absorptiometry is used to measure the total bone mineral density (BMD, g/m²) of the rats prior to their euthanasia at the end of the experiments. Then, the left femoral shafts are utilized for biomechanical testing, and the fourth lumbar vertebrae (L4) are dissected for histomorphometric and micro-computed tomographic (µCT) analysis.

[1]. Changes in gene expression induced by histamine, fexofenadine and osthole: Expression of histamine H1 receptor, COX-2, NF-κB, CCR1, chemokine CCL5/RANTES and interleukin-1β in PBMC allergic and non-allergic patients. Immunobiology . 2017 Mar;222(3):571-581.

[2]. Osthole stimulates osteoblast differentiation and bone formation by activation of beta-catenin-BMP signaling. J Bone Miner Res. 2010 Jun;25(6):1234-45.

[3]. Osthole pretreatment alleviates TNBS-induced colitis in mice via both cAMP/PKA-dependent and independent pathways. Acta Pharmacol Sin. 2017 Aug;38(8):1120-1128.

[4]. Osthole: A Review on Its Bioactivities, Pharmacological Properties, and Potential as Alternative Medicine. Evid Based Complement Alternat Med. 2015;2015:919616.

C15H16O3

244.29

244.11

C, 73.75; H, 6.60; O, 19.65

484-12-8

Osthole-d3

Solid powder

CC(=CCC1=C(C=CC2=C1OC(=O)C=C2)OC)C

MBRLOUHOWLUMFF-UHFFFAOYSA-N

InChI=1S/C15H16O3/c1-10(2)4-7-12-13(17-3)8-5-11-6-9-14(16)18-15(11)12/h4-6,8-9H,7H2,1-3H3

7-methoxy-8-(3-methylbut-2-enyl)chromen-2-one

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)

Solubility in Formulation 1: ≥ 2.5 mg/mL (10.23 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (10.23 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (10.23 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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