(Rac)-ZLc-002 is an inhibitor (blocker/antagonist) of the interaction between nNOS and nitric oxide synthase 1 adapter protein (NOS1AP), inhibits inflammatory pain and chemotherapy-induced neuropathic pain, and synergizes with Paclitaxel to reduce tumor cell viability.

Physicochemical Properties

Molecular Formula	C10H17NO5
Molecular Weight	231.25
Related CAS #	(S)-ZLc002;308277-46-5;ZLc-002;1446971-41-0
Appearance	Off-white to light yellow solid powder
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	nNOS/NOS1AP interaction[1]
ln Vitro	(Rac)-ZLc-002 (10 μM) attenuates the co-immunoprecipitation of nNOS and NOS1AP in primary cultured cortical neurons. In vitro binding assays using AlphaScreen, (Rac)-ZLc-002 was unable to break the nNOS-NOS1AP protein-protein interaction. inert in tests conducted without cells [1]. In HEK293T cells that co-express full-length nNOS, (Rac)-ZLc-002 (10 μM) can decrease the co-immunoprecipitation of full-length NOS1AP but not PSD95-PDZ2 [1]. In order to lower tumor cell viability, (Rac)-ZLc-002 (0-50 μM; 72 h) works in concert with Paclitaxel[1].
ln Vivo	In mouse models, (Rac)-ZLc-002 (10 mg/kg; ip; once or daily for 8 days) suppresses Paclitaxel-induced neuropathic pain and lessens mechanical and cold ectopic pain (HY-B0015) [1]. In the spinal dorsal horn of rats, (Rac)-ZLc-002 (4 and 10 mg/kg; ip; once) decreases formalin-induced nociceptive behavior and formalin-like immunoreactivity [1]. In ICR mice, the anxiety behavior caused by chronic mild stress (CMS) is attenuated by (Rac)-ZLc-002 (40 mg/kg/day; iv; 7 days) [2]. The behavioral effects of glucocorticoids in mice were reversed when (Rac)-ZLc-002 (10 μM; 1 μl; 30 min after Corticosterone, HY-B1618) was injected into the hippocampus for 7 days [2].
Cell Assay	Cell Viability Assay[1] Cell Types: 4T1 cells Tested Concentrations: 0-50 µM Incubation Duration: 72 h Experimental Results: Treatment alone had no effect on the viability. demonstrated synergistic effect with Paclitaxel.
Animal Protocol	Animal/Disease Models: Adult C57BL/6J male mice, paclitaxel model of neuropathic pain[1] Doses: 10 mg/kg Route of Administration: IP, once or daily for 8 days Experimental Results: Increased postinjection mechanical paw withdrawal thresholds, mechanical paw withdrawal thresholds differed across postinjection times, and the interaction between drug treatment and injection time was significant. Elevated mechanical paw withdrawal thresholds relative to vehicle treatment from 30 min. diminished postinjection cold responsiveness, cold responsiveness did not differ reliably across postinjection times, and the interaction between drug treatment and injection time was not significant. one time/day dosing increased mechanical paw withdrawal thresholds relative to the vehicle-treated group across the observation interval.
References	[1]. ZLc002, a putative small-molecule inhibitor of nNOS interaction with NOS1AP, suppresses inflammatory nociception and chemotherapy-induced neuropathic pain and synergizes with paclitaxel to reduce tumor cell viability. Mol Pain. 2018;14:1744806918801224. [2]. nNOS-CAPON blockers produce anxiolytic effects by promoting synaptogenesis in chronic stress-induced animal models of anxiety. Br J Pharmacol. 2020;177(16):3674-3690.

Solubility Data

Solubility (In Vitro)

DMSO :~100 mg/mL (~432.43 mM)

Solubility (In Vivo)

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. Injection Formulations (e.g. IP/IV/IM/SC) 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). 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) Oral Formulations 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). 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 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	4.3243 mL	21.6216 mL	43.2432 mL
	5 mM	0.8649 mL	4.3243 mL	8.6486 mL
	10 mM	0.4324 mL	2.1622 mL	4.3243 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.