Treatment with benserazide hydrochloride (BH) and levodopa (LD) alone or in combination (benserazide hydrochloride + LD) (25 μM; 0 hours, 12 hours, 24 hours, and 168 hours; SH-SY5Y) decreases protein aggregation and can limit protein aggregation. Amyloid-induced cytotoxicity in human neuroblastoma cell lines. Both benserazide hydrochloride and LD can be efficient inhibitors of the formation of cytotoxic HSA aggregates, and the inhibitory effect is more visible when the two medicines are introduced at the same time [2].

Treatment with benserazide hydrochloride (BH) and levodopa (LD) alone or in combination (benserazide hydrochloride + LD) (25 μM; 0 hours, 12 hours, 24 hours, and 168 hours; SH-SY5Y) decreases protein aggregation and can limit protein aggregation. Amyloid-induced cytotoxicity in human neuroblastoma cell lines. Both benserazide hydrochloride and LD can be efficient inhibitors of the formation of cytotoxic HSA aggregates, and the inhibitory effect is more visible when the two medicines are introduced at the same time [2].

Benserazide (5–50 mg/kg; intraperitoneal injection; male Wistar rats) treatment raised exogenous L-DOPA-derived extracellular DA levels and considerably extended the time to peak DA levels in 6-OHDA-injured rats. ..Depend on. With 10 mg/kg and 50 mg/kg benserazide, there was a substantial decrease in AADC activity in denervated striatal tissue. Benserazide alters the metabolism of exogenous L-DOPA by lowering central AADC activity in the striatum of rats suffering from nigrostriatal denervation [1].

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In sham-lesioned (intact) rats, administration of 50 mg/kg Benserazide hydrochloride (i.p.) significantly decreased endogenous extracellular dopamine (DA) levels in the striatum, with the lowest level reaching 53% of baseline at 100 minutes post-injection. Lower doses (5 and 10 mg/kg) induced a transient, non-significant decrease followed by a slight increase.[1]
In the striatum of 6-hydroxydopamine (6-OHDA)-lesioned rats (dopaminergic denervation model), pretreatment with Benserazide hydrochloride (5, 10, or 50 mg/kg, i.p., 30 minutes before L-DOPA) significantly increased the cumulative amount of extracellular DA derived from exogenous L-DOPA (50 mg/kg, i.p.) over 320 minutes compared to vehicle pretreatment.[1]
The peak extracellular DA level after L-DOPA administration was highest with 10 mg/kg Benserazide hydrochloride pretreatment. However, the time to reach the peak DA level was significantly prolonged with the 50 mg/kg dose (140 minutes) compared to the vehicle group (80 minutes) or the lower dose groups (100 minutes for 5 and 10 mg/kg).[1]
Administration of Benserazide hydrochloride (10 and 50 mg/kg, i.p.) significantly inhibited central AADC activity measured in striatal tissue homogenates from both sham-lesioned and 6-OHDA-lesioned rats 60 minutes after drug administration.[1]
In sham-lesioned rats, AADC activity was reduced to 72% of vehicle control by 10 mg/kg and to 22% by 50 mg/kg Benserazide hydrochloride.[1]
In 6-OHDA-lesioned rats, AADC activity was reduced to 25% of vehicle control by 10 mg/kg and to 12% by 50 mg/kg Benserazide hydrochloride. The remaining AADC activity in the denervated striatum (vehicle group) was about 41% of that in sham-lesioned rats.[1]

Ex Vivo AADC Activity Assay: Rats were administered Benserazide hydrochloride (10 or 50 mg/kg, i.p.) or vehicle. Sixty minutes later, they were sacrificed, and the brains were rapidly removed. The striatum was dissected on ice, homogenized in sucrose solution, and centrifuged. The supernatant was incubated at 37°C for 20 minutes in a reaction mixture containing sodium phosphate buffer (pH 7.2), pyridoxal-5-phosphate, pargyline, 2-mercaptoethanol, EDTA, ascorbic acid, and L-DOPA as the substrate. The reaction was stopped with perchloric acid containing an internal standard. After centrifugation and filtration, the amount of dopamine (DA) formed in the supernatant was quantified using High-Performance Liquid Chromatography with electrochemical detection (HPLC-ECD). AADC activity was expressed as nmol of DA formed per mg of striatal tissue per 20 minutes of incubation.[1]

Cell Viability Assay[2]
Cell Types: SH-SY5Y Cell
Tested Concentrations: 25 μM
Incubation Duration: 0 hrs (hours), 12 hrs (hours), 24 hrs (hours) and 168 hrs (hours)
Experimental Results: Enhanced cell viability and inhibited cytotoxic human serum albumin (HSA) aggregates Formation.

Animal/Disease Models: Male Wistar rat 6-hydroxydopamine (6-OHDA) (8 Ag/4 Al)[1]
Doses: 5 mg/kg, 10 mg/kg or 50 mg/kg (pharmacokinetic/PK/PK study) Give Medication: intraperitoneal (ip) injection.
Experimental Results: Extracellular DA levels derived from exogenous L-DOPA increased, and the time to reach peak DA levels was Dramatically prolonged in a dose-dependent manner. AADC activity in denervated striatal tissue was Dramatically diminished at 10 mg/kg and 50 mg/kg.

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Animal Model Preparation: Male Wistar rats (220-250 g) were used. For the dopaminergic denervation model, rats were anesthetized and a stereotaxic injection of 6-hydroxydopamine (6-OHDA, 8 µg in 4 µl) was made into the right medial forebrain bundle. Rats were pretreated with desipramine (25 mg/kg, i.p.) 30 minutes before 6-OHDA injection to protect noradrenergic neurons. Sham-lesioned rats received a saline injection. Two weeks post-surgery, successful denervation was confirmed by apomorphine-induced rotation ( >20 contralateral turns/5 min). Microdialysis experiments were performed 3-4 weeks after surgery.[1]
In Vivo Microdialysis: A guide cannula was implanted into the right striatum. A microdialysis probe was inserted and perfused with artificial Ringer’s solution at 2 µl/min. Dialysates were collected every 20 minutes. After stable baseline DA levels were established (~3 hours), Benserazide hydrochloride (5, 10, or 50 mg/kg, dissolved in saline) or vehicle was administered intraperitoneally (i.p.). In 6-OHDA-lesioned rat experiments, Benserazide hydrochloride or vehicle was administered i.p., followed 30 minutes later by L-DOPA methyl ester hydrochloride (50 mg/kg, i.p., dissolved in saline). Extracellular DA levels in dialysates were measured by HPLC-ECD.[1]
Ex Vivo Tissue Analysis for AADC Activity: Separate groups of rats (sham-lesioned and 6-OHDA-lesioned) were administered Benserazide hydrochloride (10 or 50 mg/kg, i.p.) or vehicle. Sixty minutes later, rats were decapitated, striatal tissues were rapidly dissected, and AADC activity was measured as described in the "Enzyme Assay" section.[1]

This study did not provide data on the absorption, distribution, metabolism, excretion, half-life, or oral bioavailability of benserazide hydrochloride. However, it cited an estimate from another study that the concentration of benserazide in brain tissue was approximately 1 µM after an injection of 50 mg/kg benserazide. [1]

This study did not provide specific toxicity data for benserazide hydrochloride, such as LD50, organ toxicity, drug interactions, or plasma protein binding. The decrease in endogenous dopamine levels in the striatum at high doses (50 mg/kg) observed in intact rats is a pharmacological effect related to its mechanism of action. [1]

[1]. Effects of benserazide on L-DOPA-derived extracellular dopamine levels and aromatic L-amino acid decarboxylase activity in the striatum of 6-hydroxydopamine-lesioned rats. Tohoku J Exp Med. 2003 Mar;199(3):149-59.

[2]. A multiparametric analysis of the synergistic impact of anti-Parkinson's drugs on the fibrillation of human serum albumin. Biochim Biophys Acta Proteins Proteom. 2019 Mar;1867(3):275-285.

Benserazide hydrochloride is the monohydrochloride salt of benserazide. It is an aromatic L-amino acid decarboxylase inhibitor (DOPA decarboxylase inhibitor) that does not enter the central nervous system and is therefore often used in combination with levodopa to treat Parkinson's disease. By inhibiting the peripheral conversion of levodopa to dopamine, benserazide hydrochloride increases the amount of levodopa reaching the central nervous system, thereby reducing the required dose. When used alone, benserazide hydrochloride does not have an anti-Parkinson's effect. It is both an anti-Parkinson's drug and an EC 4.1.1.28 (aromatic L-amino acid decarboxylase) inhibitor and a dopaminergic drug. It contains benserazide (1+). Benserazide hydrochloride is a peripheral aromatic L-amino acid decarboxylase (AADC) inhibitor, often used in combination with levodopa to treat Parkinson's disease by preventing the peripheral conversion of levodopa to dopamine, thereby increasing the availability of levodopa in the brain and reducing peripheral side effects. [1] This study shows that benserazide hydrochloride also has a central effect, which can exert its effect by inhibiting the activity of AADC in the brain (striatum) of intact rats and dopaminergic denervated rats (6-OHDA injury model). [1] The central inhibitory effect of benserazide hydrochloride on AADC affects the metabolism of exogenous levodopa in the denervated striatum. Peripheral inhibition can increase the bioavailability of levodopa, thereby increasing striatal dopamine levels; while central inhibition weakens the conversion of levodopa to dopamine in the brain, resulting in a longer duration of dopamine release (prolonged time to peak) at high doses (e.g., 50 mg/kg). [1] The study shows that the optimal ratio of levodopa to benserazide to maximize extracellular dopamine levels in the striatum is about 5:1 (50 mg/kg levodopa in rats: 10 mg/kg benserazide), which is consistent with clinical observations. [1] The results of the study indicate that the central activity of AADC inhibitors such as benserazide should be considered in the experimental design and clinical application of Parkinson's disease, as it can modulate the pharmacokinetics and efficacy of levodopa. [1]

C10H16CLN3O5

293.7041

293.077

14919-77-8

Benserazide;322-35-0;Benserazide-d3 hydrochloride

26964

White to off-white solid powder

574.2ºC at 760 mmHg

146°C

0.527

8

7

5

19

278

0

ULFCBIUXQQYDEI-UHFFFAOYSA-N

InChI=1S/C10H15N3O5.ClH/c11-6(4-14)10(18)13-12-3-5-1-2-7(15)9(17)8(5)16;/h1-2,6,12,14-17H,3-4,11H2,(H,13,18);1H

2-amino-3-hydroxy-N'-[(2,3,4-trihydroxyphenyl)methyl]propanehydrazide;hydrochloride

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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

DMSO : ~100 mg/mL (~340.48 mM)
H2O : ≥ 50 mg/mL (~170.24 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (7.08 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 20.8 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: ≥ 2.08 mg/mL (7.08 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 20.8 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: ≥ 2.08 mg/mL (7.08 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 100 mg/mL (340.48 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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