ML-T7 is a potent Tim-3 inhibitor. ML-T7 blocks the interaction of Tim-3 with PtdSer and CEACAM1. ML-T7 not only enhances the anti-tumor activity of CTLs and CAR T cell adoptive transfer therapy, but also enhances the effector function of T cells. ML-T7 promotes the killing activity of NK cells against tumor cells and the antigen presentation ability of DCs. ML-T7 alone or in combination with Nivolumab exerts direct anti-tumor efficacy in preclinical tumor models. ML-T7 may be utilized in tumor immunotherapy research.

Physicochemical Properties

CAS #	459789-75-4
Appearance	White to off-white 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

ln Vitro	ML-T7 (10 μM, 0–6 days) stimulates CD8+ cells' anti-tumor activity and amplifies the TCR/STAT5 signaling pathway via Tim-3 [1]. ML-T7 (10 μM, 24 h) can improve DC antigen presentation capacity and stimulate DC maturation and function via Tim-3 and Tim-4 [1]. ML-T7 (10 μM, 24 h) decreases CTL death while increasing CTL activation and cytokine production [1]. In the human NK cell line NK92 cells, ML-T7 (10 μM, 48 h) can dramatically increase the production of IFN-γ, TNF-α, CD107a, and granzyme B [1].
ln Vivo	ML-T7 (10–50 mg/kg; intraperitoneal injection; every two days, ten times) suppresses the growth of tumors and extends the life of mice without having a negative impact on their body weight [1]. The anti-tumor impact of Nivolumab antibody can be enhanced and the survival rate of HCC mice can be effectively increased by combining treatment with ML-T7 (20 mg/kg; intraperitoneal injection; every 2 days, 10 times) and Nivolumab [1]. Mouse safety results for ML-T7 (50 mg/kg, intraperitoneal injection, every two days for three weeks) are good [1].
Cell Assay	Western Blot Analysis[1] Cell Types: CD8+ cell Tested Concentrations: 10 μM Incubation Duration: 0-6 days Experimental Results: Increased the phosphorylation of phospholipase C–γ1(PLC-γ1), ZAP70, LCK, ERK1/2, and STAT5 upon anti -CD3/CD28 stimulation. Cell Invasion Assay[1] Cell Types: Bone Marrow Cells Tested Concentrations: 10 μM Incubation Duration: 24 h Experimental Results: Increased the expression of DC maturation markers.
References	[1]. Identification of a small-molecule Tim-3 inhibitor to potentiate T cell-mediated antitumor immunotherapy in preclinical mouse models. Sci Transl Med. 2023 Nov 15;15(722):eadg6752.

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.)