CSF1R, Mer (MERTK), and Axl. Adrixetinib is a triple inhibitor with Kd values of 8.7 nM (CSF1R), 0.8 nM (Mer), and 0.3 nM (Axl). It potently blocks these TAM family receptors to modulate innate immunity.

Adrixetinib has Kd values of 8.7 nM for CSF1R, 0.8 nM for Mer, and 0.3 nM for Axl. This high-affinity triple inhibition helps to repolarize tumor-associated macrophages from an immunosuppressive M2-like phenotype to a pro-inflammatory M1-like state, enhancing anti-tumor immunity and reducing tumor growth.

In preclinical mouse models, Adrixetinib has demonstrated robust anti-tumor activity by reducing the number of immunosuppressive macrophages within the tumor microenvironment and promoting T-cell activation. It has been used to validate the triple blockade of CSF1R, Mer, and Axl as a therapeutic strategy for solid tumors.

Cell-free binding affinities (Kd) are determined using a competition binding assay. Recombinant CSF1R, Mer, and Axl kinases are incubated with a proprietary ligand and varying concentrations of Adrixetinib. The degree of binding is measured using fluorescence polarization or a similar detection method, and the dissociation constant (Kd) is calculated using non-linear regression.

Murine or human macrophage cell lines (e.g., RAW 264.7 or THP-1-derived) are treated with Adrixetinib for 24-72 hours. The expression of M1 (e.g., iNOS, TNF-alpha) and M2 (e.g., Arg-1, CD206) markers is measured by qRT-PCR or flow cytometry. The ability of treated macrophages to suppress T-cell proliferation is assessed in co-culture assays.

For in vivo efficacy, syngeneic mouse tumor models (e.g., CT26 colon carcinoma or EMT6 breast cancer) are used. Adrixetinib is administered orally at 10-50 mg/kg daily. Tumor growth is monitored, and at the end of the study, tumors are analyzed by flow cytometry and immunohistochemistry to assess the percentage of M2 macrophages and the frequency of CD8+ T cells.

Adrixetinib is an orally bioavailable small molecule with a favorable ADME profile suitable for in vivo studies. Its half-life in mice and rats is sufficient to support once-daily dosing. It is metabolized in the liver primarily by CYP3A4. Its ability to penetrate the tumor microenvironment is attributed to its small molecule size (531.48 g/mol).

No specific toxicity data are detailed in the literature. As a dual CSF1R and Axl inhibitor, the primary potential toxicities are on-target and may include effects on bone metabolism (osteopetrosis) due to CSF1R inhibition. However, in preclinical studies, Adrixetinib is generally well-tolerated at therapeutic doses.

[1]. WHO Drug Informat ion - World Health Organization (WHO).

[2]. Quinoline derivatives as inhibitors of Axl/Mer RTK and CSF1R and their preparation: World Intellectual Property Organization, WO2019229251. 2019-12-05.

[3]. WHO Drug Information-World Health Organization (WHO).

Adrixetinib is a small molecule drug. The International Nonproprietary Name (INN) prefix "-tinib" in its name indicates that adrixetinib is a tyrosine kinase inhibitor. The monoisotope molecular weight of adrixetinib is 531.17 Da. Adrixetinib is an orally bioavailable receptor tyrosine kinase (RTK) inhibitor that inhibits Axl (UFO), Mer, and colony-stimulating factor-1 receptors (CSF1R; CSF-1R; CD115; M-CSFR), possessing potential immunomodulatory, chemosensitizing, and antitumor activities. After oral administration, adrixetinib targets and blocks the activity of Axl, Mer, and CSF1R, thereby blocking Axl, Mer, and CSF1R-mediated signaling pathways. This inhibits the proliferation of tumor cells expressing Axl and Mer. Furthermore, blocking Axl and Mer-mediated signaling pathways may reactivate the innate immune system against cancer cells. Blocking the CSF1R-mediated signaling pathway inhibits the activity of tumor-associated macrophages (TAMs), regulatory T cells (Tregs), and the recruitment of myeloid-derived suppressor cells (MDSCs). In summary, this can eliminate immunosuppression in the tumor microenvironment (TME), enhance anti-tumor T cell immune responses, and inhibit tumor cell proliferation. CSF1R, also known as macrophage colony-stimulating factor receptor (M-CSFR) and CD115 (differentiation cluster 115), is a cell surface receptor that plays a crucial role in tumor cell proliferation, metastasis, and cancer-related immunosuppression. Axl and Mer are both members of the TAM (Tyro3, Axl, and Mer) family of RTKs and are overexpressed in various tumor cell types. They play key roles in tumor cell proliferation, survival, invasion, angiogenesis, metastasis, and inactivation of innate immunity in cancer. Their expression is associated with drug resistance and poor prognosis.

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Other information: Adrixetinib is a cutting-edge research tool in immuno-oncology. It is used to dissect the roles of TAM receptors in immunosuppression and has potential for combination therapy with immune checkpoint inhibitors. It is not an FDA-approved drug and is strictly for research use only (CAS: 2394874-66-7).

C25H24F3N5O5

531.483776092529

531.172

2394874-66-7

Adrixetinib TFA

156440741

White to off-white solid powder

4.3

1

11

10

38

765

0

N1(CCC)C=C(OCC(F)(F)F)C(C(NC2=NC=C(OC3C4C(N=CC=3)=CC(OC)=C(OC)C=4)C=C2)=O)=N1

BWUUJXXFCSYLFW-UHFFFAOYSA-N

InChI=1S/C25H24F3N5O5/c1-4-9-33-13-21(37-14-25(26,27)28)23(32-33)24(34)31-22-6-5-15(12-30-22)38-18-7-8-29-17-11-20(36-3)19(35-2)10-16(17)18/h5-8,10-13H,4,9,14H2,1-3H3,(H,30,31,34)

N-[5-(6,7-dimethoxyquinolin-4-yl)oxypyridin-2-yl]-1-propyl-4-(2,2,2-trifluoroethoxy)pyrazole-3-carboxamide

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ≥ 100 mg/mL (188.15 mM)

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.)