ZLMT-12 (compound 35) is a tacnin analogue and a potent CDK2/9 inhibitor. The IC50s for inhibiting CDK9 and CDK2 are 0.002 and 0.011 μM respectively. ZLMT-12 has a weak inhibitory effect on AChE (IC50=19.023 μM) and BuChE (IC50=2.768 μM). ZLMT-12 has low toxicity and antiproliferation activity. ZLMT-12 causes apoptosis and blocks the cell cycle in S phase and G2/M phase.

Physicochemical Properties

CAS #	2841473-39-8
Appearance	Typically exists as solid at room temperature
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	CDK9 0.002 μM (IC50) CDK2 0.011 μM (IC50) BChE 2.768 μM (IC50) AChE 19.023 μM (IC50)
ln Vitro	Compound 35, ZLMT-12, exhibits antiproliferative action in cancer cells at 500 nM for 72 hours[1]. ZLMT-12 (500 nM; 72 h) stops the cell cycle in the S and G2/M phases and causes apoptosis[1].
ln Vivo	In the HCT116 xenograft model, ZLMT-12 (compound 35; 10 mg/kg; po; daily, for 21 d) demonstrates antitumor efficaciousness and reduces tumor growth, without causing injury to the liver[1].
Cell Assay	Cell Viability Assay[1] Cell Types: HCT116, SW480, A549, and MCF-7 cells Tested Concentrations: 500 nM Incubation Duration: 72 hrs (hours) Experimental Results: Inhibited cell proliferative with GI50 values of 0.029, 0.328, 0.051, and 0.109 μM for HCT116, SW480, A549, and MCF-7 cells, respectively. Apoptosis Analysis[1] Cell Types: HCT116 cells Tested Concentrations: 10 and 20 nM Incubation Duration: 48 hrs (hours) Experimental Results: Increased apoptotic cells rate from 9.22% in the control to 23.77 % at 10 nM and increased apoptotic cells rate to 46.2% at 20 nM. Cell Cycle Analysis[1] Cell Types: HCT116 cells Tested Concentrations: 10 and 20 nM Incubation Duration: 48 hrs (hours) Experimental Results: Increased the percentage of the S phase from 31.43% to 42.75% (10 nM) and 49.38% (20 nM) respectively, and the percentage of the G2/M phase from 6.39% to 10.60% (10 nM) and 13.11% (20 nM), respectively.
Animal Protocol	Animal/Disease Models: Male BALB/cA-nu (nude) mice with HCT116 xenografts (18-25 g, 6-8 weeks of age)[1] Doses: 10 mg/kg Route of Administration: Oral administration; daily, for 21 days Experimental Results: Inhibited tumor growth with GI (tumor volume growth inhibition)= 47.66% and TGI (tumor weight growth inhibition)=62.39%. demonstrated no significant changes in behavior or body weight in mice. Had no obvious liver injury. Animal/Disease Models: Male SD (Sprague-Dawley) rats (240±20 g)[1] Doses: 2 mg/kg (iv) and 20 mg/kg (po) (pharmacokinetic/PK Analysis) Route of Administration: intravenous (iv) injection and oral administration; once Experimental Results: 1.19 Parameter 2 mg/kg (iv) 20 mg/kg (po) T1/2 (h) 0.461 1.77 Tmax (h) 0.083/td> 1.00 Cmax (ng/mL) 206 67.8 AUCo-t (h*ng/mL) 110 302 AUCo-∞ (h*ng/mL) 115 316 CL (L/ h/kg) 18.7 Vss(L/Kg) 12.6 F (%) 27.47
References	[1]. Development and structure-activity relationship of tacrine derivatives as highly potent CDK2/9 inhibitors for the treatment of cancer. Eur J Med Chem. 2022 Nov 15;242:114701.

Solubility Data

Solubility (In Vitro)

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

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.)