Compound 24 (3-Methyl-6-(2-(4-(2-methoxyphenyl)piperazin-1-yl)ethyl)benzothiazolin-2-one) is a multi-receptor ligand. The half-maximal inhibitory concentrations (IC₅₀) from receptor binding assays are: 5-HT_1A (2 ± 0.3 nM), 5-HT_1B (300 ± 55 nM), 5-HT_2A (500 ± 75 nM), 5-HT_2C (4000 ± 440 nM), D₂ (40 ± 9 nM), and α₁ (10 ± 3 nM). It functions as a 5-HT_1A ligand, a D₂ antagonist, and a 5-HT_2A antagonist [1].

5-HT1A modulator 1 (compound 24) exhibits affinity for 5HT1B, 5-HT2A, and 5-HT2C receptors, with IC50 values of 300±55 nM, 500±75 nM, and 4000±440 nM, in that order [1].

Compound 24, also known as 5-HT1A modulator 1, had notable antagonistic effects on the mouse 5HT2A receptor subtype. At 1 mg/kg ip dosage, 5-HT1A Modulator 1 antagonistic activity was nearly full (94% antagonism, p<0.01). At an intraperitoneal dose of 1 mg/kg, 5-HT1A modulator 1 totally prevented setting and rising (100% antagonism). Rats were also used to test 5-HT1A modulator 1 using the same paradigm. 5-HT1A modulator 1 significantly (p<0.05) decreased ADHD after oral dosing at oral dosages of 2 and 4 mg/kg, which represented 63% and 58% of 5-HT1A modulator 1's antagonism, respectively. Complete (103% and 108%), 5-HT1A modulator 1 antagonistic effects were observed at oral dosages of 8 and 16 mg/kg (p<0.01) [1].

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Compound 24 (1 mg/kg, ip) significantly antagonized the hyperactivity induced by d-amphetamine in mice, showing 57% antagonism (p < 0.01), which was comparable to the effects of clozapine and haloperidol [1].
In rats, after oral administration (po), compound 24 at doses of 2 mg/kg and 8 mg/kg po reduced the d-amphetamine-induced hyperactivity by 63% (p < 0.05) and completely blocked it (103% antagonism, p < 0.01), respectively [1].
Compound 24 (0.25 and 1 mg/kg, ip) potently and significantly (p < 0.001) antagonized mescaline-induced scratching in mice, with 92% and 100% antagonism, respectively, indicating serotonergic antagonism [1].
Compound 24 (1 mg/kg, ip) completely blocked the stereotypies and climbing induced by a low dose of apomorphine in mice (100% antagonism), demonstrating potent D₂ antagonist activity [1].
Compound 24 showed a dose-dependent antagonism of the hyperlocomotion induced by the σ agonist SKF 10047 in rats (39%, 56%, 71% antagonism at 0.5, 2, 8 mg/kg po, p < 0.05 to p < 0.001) [1].
In the tail suspension test in mice, compound 24 (1 mg/kg, ip) increased the duration of immobility by 89% (p < 0.001), a profile consistent with neuroleptic drugs [1].
Compound 24 did not induce forepaw treading in rats at doses active in antipsychotic models, indicating a lack of in vivo 5-HT_1A agonist-like effects [1].
Compound 24 (2 mg/kg, ip) only weakly increased the duration of barbital-induced sleep in mice (50% increase, not significant), indicating low sedative potential at behaviorally active doses [1].
In the Sidman avoidance test in rats, compound 24 dose-dependently decreased the number of avoidance responses after oral administration. At 32 mg/kg po, it decreased responses by 78% (p < 0.001) and increased the number of shocks and escape failures significantly, indicating neuroleptic activity [1].
Compound 24 did not induce catalepsy in rats at pharmacologically active doses (1 mg/kg, ip). Significant catalepsy was only observed at a very high dose (32 mg/kg, ip), suggesting a low potential for inducing extrapyramidal side effects (EPS) [1].

5-HT_1A Receptor Binding Assay: The assay was performed using membranes prepared from bovine hippocampus. The receptor was labeled with 0.5 nM [³H]-8-OH-DPAT. Incubation was carried out at 25°C for 30 minutes in the presence of 11 concentrations of the test compound (10^-9 to 10^-4 M). Nonspecific binding was determined using 10^-5 M buspirone. Competition data were analyzed using a nonlinear curve-fitting program to calculate IC₅₀ values [1].
D₂ Receptor Binding Assay: The assay was performed using membranes prepared from bovine striatum. The receptor was labeled with 1.2 nM [³H]raclopride. Incubation was carried out at 25°C for 30 minutes in the presence of 11 concentrations of the test compound (10^-9 to 10^-4 M). Nonspecific binding was determined using 10^-5 M spiperone. Competition data were analyzed to calculate IC₅₀ values [1].
5-HT_2A Receptor Binding Assay: The assay was performed using membranes prepared from bovine frontal cortex. The receptor was labeled with 0.8 nM [³H]ketanserin. Incubation was carried out for 30 minutes at 37°C. Nonspecific binding was determined using 10^-5 M spiperone. Competition data were analyzed to calculate IC₅₀ values [1].
α₁ Receptor Binding Assay: The assay was performed using membranes prepared from bovine frontal cortex. The receptor was labeled with 0.5 nM [³H]prazosin. Incubation was carried out at 25°C for 40 minutes in the presence of 11 concentrations of the test compound (10^-9 to 10^-4 M). Nonspecific binding was determined using 10^-5 M phentolamine. Competition data were analyzed to calculate IC₅₀ values [1].

d-Amphetamine-induced Hyperactivity in Mice: Swiss mice were pretreated with d-amphetamine (4 mg/kg, intraperitoneal, ip). Vehicle or the test compound was administered 30 minutes after amphetamine. Locomotor activity was then recorded for 30 minutes in plexiglass chambers. Statistical comparison was made using Student's t-test [1].
d-Amphetamine-induced Hyperactivity in Rats: Wistar rats were pretreated with d-amphetamine (4 mg/kg, ip). The test compound was administered orally (po) in specified doses. Locomotor activity was recorded for 30 minutes. Statistical comparison was made using Student's t-test [1].
Mescaline-induced Scratching in Mice: Swiss mice were injected with mescaline hydrochloride (25 mg/kg, ip) and placed in square cages. The number of scratching paroxysms (defined as brief bursts of scratching the head/ear with the hind foot) was counted for 5 minutes, starting 30 minutes after treatment. Results were calculated as percent antagonism of scratching compared to controls [1].
Apomorphine Antagonism in Mice: The intensity of stereotypies, climbing behavior, and rectal temperature were measured in mice 30 minutes after administration of apomorphine (1 mg/kg, subcutaneous, sc). The test compound was administered prior to apomorphine to evaluate D₂ antagonism [1].
Tail Suspension Test in Mice: Swiss mice were suspended by the tail for 6 minutes. Their behavior was recorded automatically using a computerized apparatus. The duration of immobility was measured. Neuroleptics typically increase immobility time [1].
Sidman Avoidance Test in Rats: Lister Hooded rats, pretrained in a Skinner box, were required to press a lever at regular intervals to avoid an electric shock. After administration of the test compound (po), the number of avoidance responses, shocks received, and escape failures were recorded. The compound's effect on conditioned avoidance behavior was evaluated [1].
Catalepsy Test in Rats: Wistar rats were administered the test compound or vehicle and tested for catalepsy at 30-minute intervals for 90 minutes. Catalepsy was assessed by three methods: imposed crossing of ipsilateral fore- and hindlimbs, placing in the "Buddha" position, and using a tilting board device [1].

[1]. Novel benzothiazolin-2-one and benzoxazin-3-one arylpiperazine derivatives with mixed 5HT1A/D2 affinity as potential atypical antipsychotics. J Med Chem. 1998 Jun 4;41(12):2010-8.

Compound 24 is a benzothiazolin-2-one derivative containing an arylpiperazine moiety. It was designed as a mixed 5-HT1A/D2 ligand based on the assumption that such ligand properties might confer atypical antipsychotic activity and reduce extrapyramidal side effects (EPS) [1]. The compound exhibits multi-receptor binding properties with high affinity for both 5-HT1A and D2 receptors. In vivo experiments showed that the compound exhibited potent antipsychotic-like activity in a variety of predicted models (e.g., antagonism of amphetamine- and mescaline-induced behavior) with a low tendency to induce rigidity at pharmacologically relevant doses. This combination of high efficacy and low EPS risk makes it a promising potential atypical antipsychotic in this study [1].

C21H25N3O2S

383.507103681564

383.166

142477-34-7

9821397

White to off-white solid powder

3.7

0

5

5

27

512

0

S1C2=CC(CCN3CCN(C4=CC=CC=C4OC)CC3)=CC=C2N(C)C1=O

RTHIBOYVHBRSHH-UHFFFAOYSA-N

InChI=1S/C21H25N3O2S/c1-22-18-8-7-16(15-20(18)27-21(22)25)9-10-23-11-13-24(14-12-23)17-5-3-4-6-19(17)26-2/h3-8,15H,9-14H2,1-2H3

6-[2-[4-(2-methoxyphenyl)piperazin-1-yl]ethyl]-3-methyl-1,3-benzothiazol-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)

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.)