PF-07245303 (0.49-40 nM; binding period 60 seconds, dissociation period 60-480 seconds) can effectively bind recombinant human ITK, TRKA, TRKB and TRKC proteins, with Kd values of 0.71 nM, 2.94 nM, 8.73 nM and 10 nM, respectively [1]. Under 1 mM ATP conditions, PF-07245303 can effectively inhibit the enzyme activity of full-length human ITK protein, with an IC50 value of 5.62 nM [1]. PF-07245303 (0-10 μM; pre-incubation for 30 minutes, activation for 5 minutes) can effectively inhibit ITK-dependent PLCγ1 phosphorylation in Jurkat T cells activated by anti-CD3/CD28 antibody, with an IC50 of 154 nM [1]. PF-07245303 can effectively inhibit the release of IL-2 from human primary CD4+ T cells activated by anti-CD3/CD28 antibodies, with an IC50 of 38.5 nM[1]. PF-07245303 can inhibit the release of IL-2 from human whole blood T cells activated by anti-CD3/CD28/CD2 antibodies, with an IC50 of 1350 nM (122 nM after protein binding correction)[1]. PF-07245303 (20-24 hours) inhibits the release of IL-4, IL-13, IFNγ, IL-17A and IL-10 from human primary CD4+ T cells activated by anti-CD3/CD28, with corresponding IC50 values of 73.5 nM, 348 nM, 73.2 nM, 282 nM and 114.5 nM, respectively[1]. PF-07245303 (6-7 days) inhibited the release of cytokines from differentiated human Th1, Th2 and Th17 cells, with IC50 values of 35.4 nM, 60.9 nM and 12.5 nM for IFNγ, IL-13 and IL-17A, respectively [1]. PF-07245303 effectively inhibited the release of IFNγ from human primary CD8+ T cells activated by anti-CD3/CD28 antibody, with an IC50 value of 18.9 nM [1]. PF-07245303 effectively inhibited the enzyme activity of the cytoplasmic domains of human TRKA, TRKB and TRKC, with average IC50 values of 7.2 nM, 12.0 nM and 20.7 nM, respectively, under 1 mM ATP conditions [1]. PF-07245303 inhibited ligand-induced phosphorylation of human TRKA, TRKB, and TRKC in U2OS cells (expressing p75) with IC50 values of 72.6 nM, 48.7 nM, and 21.7 nM, respectively [1]. PF-07245303 inhibited NGF-induced activation of primary human peripheral blood basophils with an IC50 of 442 nM (39.8 nM after protein binding correction) [1]. PF-07245303 (0.1-10 μM; IL-4/IL-13 pretreatment for 24 hours, NGF co-incubation for 5 minutes) inhibited NGF-induced TRKA phosphorylation in human skin explants pretreated with IL-4/IL-13 [1]. PF-07245303 (0.3-10 μM; 24 hours) inhibited the expression of anti-CD3/CD28-induced IFNG, IL13 and IL2 mRNA in a concentration-dependent manner, with almost complete inhibition at 10 μM; at the same time, the drug could also inhibit the release of IL-2 protein, with significant inhibitory effects detected at concentrations of 1, 3 and 10 μM [1].

PF-07245303 (2%; topical application; once daily) significantly reduced oxazolone-induced ear swelling, disease severity score, and pro-inflammatory cytokine levels in mice, with an ear thickness inhibition rate of 43%-61% observed at multiple time points [1].

Western Blot Analysis [1]
Cell Types: Jurkat T cells (clone E6-1)
Tested Concentrations: 0, 0.001, 0.003, 0.01, 0.03, 0.1, 1, 3.2, 10 μM
Incubation Duration: 30 minutes (pre-incubation); 5 minutes (activation)
Experimental Results: Inhibited anti-CD3/CD28-induced PLCγ1 phosphorylation in a concentration-dependent manner, with an IC50 of 154 nM.

Animal/Disease Models:BALB/c (female, 8-10 weeks old, oxazolone-induced contact hypersensitivity model) [1]
Doses: 2%
Route of Administration: Topical application; once daily; administration started 1 hour after the first exposure to oxazolone, and was administered on days 4, 7, 9 and 11 after exposure to oxazolone stimulation.
Experimental Results: On days 4, 7, 9 and 11 after exposure to oxazolone, auricular thickening was inhibited by 43%-61%. Visual and histological comprehensive disease scores were reduced compared with the solvent group. Protein levels of IL-4, IL-10, IL-17A, GM-CSF and TNFα in ear tissue were moderately reduced. There was no difference in protein levels of IL-2 and IFNγ in ear tissue compared with the control group. Scores for dermal inflammatory cell infiltration and epithelial endpoints (including hyperplasia, ulceration, thinning, and surface hyperkeratosis) were moderately reduced.

[1]. Discovery of an ITK and TRK kinase inhibitor for the potential topical treatment of atopic dermatitis. Nat Commun. Published online March 9, 2026.

C25H32N6O2

448.56

2655556-75-3

Typically exists as solids at room temperature

CC1=C(C=C2NC(C3=NNC4=C3C[C@@H]5C[C@@]5(C4)C)=NC2=C1)N(C([C@@H](N6CCOCC6)C)=O)C

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.)