TMP195 (TMP-195) is a first-in-class and selective inhibitor of class IIa histone deacetylase (HDAC) with anticancer and immunomodulatory effects. Its IC50 value for HDAC inhibition is 300 nM. By modifying macrophage phenotypes, in vivo TMP195 treatment modifies the tumor microenvironment and lowers tumor burden and pulmonary metastases. Within tumors, TMP195 promotes the recruitment and differentiation of highly stimulatory and phagocytic macrophages. Moreover, in this model, the combination of TMP195 and chemotherapy regimens or T-cell checkpoint blockade greatly improves the durability of tumor reduction. These findings present class IIa HDAC inhibition as a way to improve cancer therapy by utilizing macrophages' anti-tumor properties. In the supernatants of cultures where monocytes are differentiated into macrophages, TMP195 prevents the buildup of CCL2 protein. Comparing the vehicle group to the TMP195 group, the monocytes secrete significantly more CCL1 protein. TMP195 down- or up-regulates CCL2 and CCL1, respectively, according to the transcriptional profiling data from the PHA-stimulated PBMC experiments.

Physicochemical Properties

Molecular Formula	C23H19F3N4O3
Molecular Weight	456.42
Exact Mass	456.14
Elemental Analysis	C, 60.53; H, 4.20; F, 12.49; N, 12.28; O, 10.52
CAS #	1314891-22-9
Related CAS #	1314891-22-9
PubChem CID	67324851
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Index of Refraction	1.546
LogP	5.57
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	6
Heavy Atom Count	33
Complexity	672
Defined Atom Stereocenter Count	0
SMILES	FC(C1=NC(C2C([H])=C([H])C([H])=C(C=2[H])C(N([H])C([H])([H])C(C([H])([H])[H])(C([H])([H])[H])C2=C([H])OC(C3C([H])=C([H])C([H])=C([H])C=3[H])=N2)=O)=NO1)(F)F
InChi Key	QTCSXAUJBQZZSN-UHFFFAOYSA-N
InChi Code	InChI=1S/C23H19F3N4O3/c1-22(2,17-12-32-20(28-17)14-7-4-3-5-8-14)13-27-19(31)16-10-6-9-15(11-16)18-29-21(33-30-18)23(24,25)26/h3-12H,13H2,1-2H3,(H,27,31)
Chemical Name	N-[2-methyl-2-(2-phenyl-1,3-oxazol-4-yl)propyl]-3-[5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl]benzamide
Synonyms	TFMO 2; TMP-195; TMP195; TMP 195
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	HDAC9 ( IC50 = 9 nM ); HDAC7 ( IC50 = 46 nM ); HDAC5 ( IC50 = 106 nM ); HDAC4 ( IC50 = 111 nM ); HDAC8 ( IC50 = 11700 nM ); HDAC6 ( IC50 = 47800 nM )
ln Vitro	In vitro activity: TMP195 fully inhibits class IIa HDAC activity without inhibiting other HDACs due to its low potency in recombinant class I and IIb HDAC assays. TMP195 dramatically increases the amount of CCL1 protein secreted by the monocytes in comparison to vehicle (DMSO)-treated M-CSF plus GM-CSF cultures, and it inhibits the accumulation of CCL2 protein in the supernatants of monocyte-derived macrophage differentiation cultures[1]. In vitro, TMP195 affects how human monocytes react to colony-stimulating factors CSF-1 and CSF-2[2].
ln Vivo	TMP195 treatment in vivo modifies macrophage phenotypes, changing the tumor microenvironment and lowering tumor burden and pulmonary metastases. Highly phagocytic and stimulatory macrophages are recruited and differentiated within tumors by TMP195. Proliferating tumor cells are dramatically reduced by TMP195 treatment, especially those near the tumor's leading edge. In this mouse model of breast cancer, checkpoint blockade immunotherapy and standard chemotherapy regimens are both more effective and durable due to the anti-tumour macrophage phenotype induced by TMP195 treatment[2].
Enzyme Assay	The labeled recombinant HDAC7 catalytic domain (amino acids 483-903) is applied to an arrayed library of 3,868 immobilized 20-mer peptides using DyLight 650. Using an automated TECAN HS4 microarray processing station, arrays are performed. First, blocking buffer is incubated for 30 minutes at 30°C. Next, saline containing 50 mM Tris Base and 0.1% Tween-20 (pH 7.2) is ished. Finally, the labeled HDAC7 protein is incubated for 120 minutes at 4°C. The labeled protein is pre-incubated with TMP195 for 30 minutes prior to application to the array in TMP195 competition experiments. After that, the microarrays are ished, dried, and imaged using a scanner.
Cell Assay	For five days, monocytes were differentiated into antigen-presenting cells in RPMI Medium 1640 supplemented with either 0.1% (v/v) DMSO or 300 nM TMP195. Other supplements included IL-4 (10 ng/ml), penicillin (100 U/ml), streptomycin (100 μg/ml), and GlutaMAX fetal bovine serum (10% v/v). After washing and incubating the cells in a 5 mM EDTA solution in Ca2+/Mg2+-free PBS, the cells were collected for flow cytometric analysis.
Animal Protocol	Mice: In every mouse study, 50 μL of the vehicle, dimethyl sulfoxide (DMSO), or 50 μL of TMP195 dissolved in 100% DMSO at a final concentration of 50 mg per kg daily, are injected intraperitoneally (i.p.).
References	[1]. Selective class IIa histone deacetylase inhibition via a nonchelating zinc-binding group. Nat Chem Biol. 2013 May;9(5):319-25. [2]. Class IIa HDAC inhibition reduces breast tumors and metastases through anti-tumor macrophages. Nature. 2017 Mar 16;543(7645):428-432.

Solubility Data

Solubility (In Vitro)

DMSO: 91~100 mg/mL (199.4~219.1 mM) Water: <1 mg/mL Ethanol: ~91 mg/mL (~199.4 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 3 mg/mL (6.57 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 30.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 3 mg/mL (6.57 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 30.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 3 mg/mL (6.57 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 30.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.. Solubility in Formulation 4: 5%DMSO+ Corn oil: 5.0mg/ml (10.95mM)  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1910 mL	10.9548 mL	21.9096 mL
	5 mM	0.4382 mL	2.1910 mL	4.3819 mL
	10 mM	0.2191 mL	1.0955 mL	2.1910 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.