| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
IC50: GLP-1 receptor
GLP-1 receptor (orthosteric agonist). [1] β-endorphin (indirectly, via stimulating expression in microglia). [1] μ-opioid receptor (indirectly, via β-endorphin release). [1] |
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| ln Vitro |
In primary cultures of rat microglial cells, incubation with Shanzhiside methylester (300 μM) significantly increased the level of β-endorphin in the culture medium by 1.3 times (2 hours after incubation). This effect was completely suppressed by pre-treatment (1 hour prior) with minocycline (60 μM). Shanzhiside methylester did not stimulate β-endorphin release in primary cultures of neurons or astrocytes. [1]
In primary rat microglial cells, Shanzhiside methylester (300 μM) significantly increased the POMC (proopiomelanocortin, the β-endorphin precursor) mRNA expression by 3.1 times (2 hours after incubation). This effect was completely inhibited by minocycline. Shanzhiside methylester did not significantly increase POMC gene expression in primary cultures of neurons or astrocytes. [1] In primary rat microglial cells, Shanzhiside methylester (300 μM) significantly increased the POMC mRNA expression by 9.5 times (2 hours after incubation). Pre-treatment (1 hour prior) with the selective p38 MAPK activation inhibitor SB203580 (50 μM) completely prevented this increase. In contrast, pre-treatment with the JNK MAPK inhibitor SP600125 (50 μM) or the ERK1/2 inhibitor U0126 (50 μM) did not significantly attenuate the Shanzhiside methylester-increased POMC expression. [1] |
| ln Vivo |
Methyl ester of lobeside, with an estimated ED50 of 40.4 μg, and a maximum inhibition rate of 49%, exhibits a dose-dependent and long-lasting (>4 h) anti-allodynic action on rats with neuropathy produced by spinal cord nerve injury [1].
In spinal nerve ligation-induced neuropathic rats, intrathecal injection of Shanzhiside methylester (10, 30, 100, and 300 μg) elevated ipsilateral paw withdrawal thresholds in a time- and dose-dependent manner without affecting contralateral paw withdrawal thresholds. The Emax was 49% MPE (Maximal Possible Effect) and the projected ED50 was 40.4 μg (95% confidence limits: 32.4 - 50.4 μg). The anti-allodynic effect lasted for >4 hours. This effect was completely inhibited by the selective GLP-1 receptor antagonist exendin(9-39) (2 μg). [1] In neuropathic rats, intrathecal injection of Shanzhiside methylester (100 μg) significantly increased β-endorphin levels by 0.8 and 1.1 times in the homogenates of the contralateral and ipsilateral spinal cords, respectively (0.5 hours after injection). This increase was completely inhibited by minocycline (100 μg). [1] The anti-allodynic effect of intrathecal Shanzhiside methylester (100 μg) in neuropathic rats was completely blocked by pre-treatment (0.5 hours earlier) with β-endorphin antiserum (1:10 dilution), the selective μ-opioid receptor antagonist CTAP, the microglial inhibitor minocycline (100 μg), and the p38 MAPK inhibitor SB203580 (30 μg). Pre-treatment with κ-opioid receptor antagonist nor-BNI or δ-opioid receptor antagonist natrlindole did not significantly inhibit the effect. [1] In neuropathic rats, repeated bi-daily intrathecal injections of Shanzhiside methylester (100 μg) over 7 days did not induce self-tolerance to its anti-allodynic effect nor cross-tolerance to morphine anti-allodynia. Concurrent treatment with Shanzhiside methylester and morphine for 7 days did not diminish the anti-allodynic effect of a subsequent morphine challenge. However, Shanzhiside methylester was not effective in relieving mechanical allodynia in morphine-tolerant rats. [1] Shanzhiside methylester (100 μg, intrathecal) and exenatide (100 ng, intrathecal) specifically activated spinal p38 MAPK phosphorylation in neuropathic rats, without affecting phospho-ERK1/2 or phospho-JNK signaling. [1] |
| Cell Assay |
Primary microglial cells were derived from 1-day-old neonatal rats. The cells were plated in a 75-cm² tissue culture flask (1×10⁷ cells/flask) coated with poly-L-lysine. After an 8-day culture, microglial cells were prepared as floating cell suspensions by shaking the flask at 260 rpm for 2 hours. The harvested microglia exhibited a purity >95% as determined by CD11b (OX42) immunoreactivity. For the experiment, Shanzhiside methylester (300 μM) was incubated with primary microglial cells. The culture medium and primary cells were collected 2 hours after incubation to measure β-endorphin levels and POMC gene expression, respectively. Minocycline (60 μM), SB203580 (50 μM), U0126 (50 μM), or SP600125 (50 μM) were incubated for 1 hour before Shanzhiside methylester treatment. [1]
Primary neuronal and astrocyte cultures were also used. For neurons, cell suspensions (1×10⁵ cells/well) were plated in a 24-well plate coated with poly-L-lysine. The medium was changed to Neurobasal medium containing B27 supplement, and experiments were initiated 8-16 days later (purity >90% by NeuN immunoreactivity). For astrocytes, after microglial removal, the nearly intact layer of astrocytes was trypsinized and subcultured (purity >90% by GFAP immunoreactivity). Shanzhiside methylester (300 μM) was incubated, and β-endorphin release and POMC expression were measured. [1] |
| Animal Protocol |
Male Wistar adult rats (160-250 g) were used. For intrathecal catheterization, rats were anesthetized with isoflurane (4% for induction, 1% for maintenance). A polyethylene catheter (PE-10, 13-μl volume) was inserted into the spinal cord at the lumbar level. Catheter placement was verified by intrathecal administration of 4% lidocaine (10 μl). Only rats exhibiting bilateral paralysis of hindlimbs after lidocaine injection were selected. Intrathecal injections were performed with a 50-μl micro-injector, injecting 10 μl of the test article followed by a 15 μl normal saline flush. [1]
To induce neuropathy, unilateral ligation of the left L5 and L6 spinal nerves was performed under anesthesia immediately after intrathecal catheterization. The nerves were isolated and tightly ligated with 6-0 silk thread. Drug tests started 1-2 weeks after surgery. Only rats with marked unilateral allodynia (hindlimb withdrawal thresholds <10 g on the operated side) and no major motor impairment were selected. [1] For the dose-response study, neuropathic rats received a single intrathecal injection of saline (10 μl) or Shanzhiside methylester at doses of 10, 30, 100, and 300 μg. For tolerance studies, rats received bi-daily intrathecal injections (twice a day) of saline, morphine (20 μg), β-endorphin (1 μg), Shanzhiside methylester (100 μg), exenatide (100 ng), or combinations for 7 days. On the eighth day, they received a challenge injection of the respective drugs. For antagonist studies, exendin(9-39) (2 μg), β-endorphin antiserum (1:10 dilution), CTAP, nor-BNI, natrlindole, minocycline (100 μg), SB203580 (30 μg), U0126 (20 μg), or SP600125 (30 μg) were intrathecally injected 0.5 or 4 hours (minocycline) before Shanzhiside methylester (100 μg) treatment. [1] Mechanical allodynia was assessed using an electrical von Frey hair (forces ranging from 0.1 to 90 g). The monofilament was applied to the foot pad with increasing force until the rat withdrew its hindlimb. The lowest force producing a withdrawal response was considered the threshold. Triplicate measurements were made at approximately 30-second intervals, and the three values were averaged for each hindpaw at each time point. [1] |
| References | |
| Additional Infomation |
methyl benzyl ester is a glycoside compound belonging to the iridoid Monotropein class. It has been reported to exist in Malus baccata, Gardenia jasminoides, and other organisms with relevant data.
Shanzhiside methylester is a small molecule (molecular weight approximately 390 Da) orthosteric agonist of the GLP-1 receptor. It is a principle effective iridoid glycoside from the analgesic herb Lamiophlomis rotata, which is listed in the Pharmacopoeia of China and has been used for hundreds of years to manage cancer pain, postoperative and bone fracture pain, and neuropathic pain. [1] Unlike morphine or exogenous β-endorphin, chronic treatment with Shanzhiside methylester does not induce anti-nociceptive tolerance. This suggests that stimulating endogenous β-endorphin expression may be a novel strategy for developing analgesics for long-term treatment of chronic pain without the risk of tolerance associated with direct μ-opioid receptor agonists. [1] |
| Molecular Formula |
C17H26O11
|
|---|---|
| Molecular Weight |
406.3817
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| Exact Mass |
406.147
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| CAS # |
64421-28-9
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| PubChem CID |
13892722
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| Appearance |
White to off-white solid
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| Density |
1.6±0.1 g/cm3
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| Boiling Point |
651.4±55.0 °C at 760 mmHg
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| Melting Point |
90ºC (dec.)
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| Flash Point |
233.0±25.0 °C
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| Vapour Pressure |
0.0±4.4 mmHg at 25°C
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| Index of Refraction |
1.621
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| LogP |
-3.25
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| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
11
|
| Rotatable Bond Count |
5
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| Heavy Atom Count |
28
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| Complexity |
624
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| Defined Atom Stereocenter Count |
10
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| SMILES |
O([H])[C@@]1(C([H])([H])[H])C([H])([H])[C@]([H])([C@]2([H])C(C(=O)OC([H])([H])[H])=C([H])O[C@]([H])([C@]12[H])O[C@@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[H])O[H])O[H]
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| InChi Key |
KKSYAZCUYVRKML-IRDZEPHTSA-N
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| InChi Code |
InChI=1S/C17H26O11/c1-17(24)3-7(19)9-6(14(23)25-2)5-26-15(10(9)17)28-16-13(22)12(21)11(20)8(4-18)27-16/h5,7-13,15-16,18-22,24H,3-4H2,1-2H3/t7-,8-,9+,10-,11-,12+,13-,15+,16+,17+/m1/s1
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| Chemical Name |
methyl (1S,4aS,5R,7S,7aS)-5,7-dihydroxy-7-methyl-1-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4a,5,6,7a-tetrahydro-1H-cyclopenta[c]pyran-4-carboxylate
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~246.08 mM)
H2O : ~50 mg/mL (~123.04 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.15 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.15 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.15 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 50 mg/mL (123.04 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4608 mL | 12.3038 mL | 24.6075 mL | |
| 5 mM | 0.4922 mL | 2.4608 mL | 4.9215 mL | |
| 10 mM | 0.2461 mL | 1.2304 mL | 2.4608 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.
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