TAK-418 is a potent and selective inhibitor of lysine-specific demethylase 1 (LSD1/KDM1A) enzyme activity. It inhibits LSD1 with an IC₅₀ of 2.9 nM and a \(k_{\text{inact}}/K_{\text{I}}\) value of \(3.8 \times 10^5 \pm 3.8 \times 10^4\) M⁻¹ s⁻¹ [1].

In primary cultured rat neurons, treatment with TAK-418 (0.0001–10 µM) for 3 days concentration-dependently increases H3K4me2 and H3K9me2 levels at the Ucp2 gene and induces Ucp2 mRNA expression, with significant effects observed at 0.01 µM [1].
TAK-418 has minimal impact on the interaction between LSD1 and its cofactor GFI1B in TF-1a cells, unlike the conventional LSD1 inhibitor T-711. It forms a compact formylated adduct with the FAD cofactor in LSD1, avoiding steric interference with GFI1B binding [1].

TAK-418 (1 mg/kg; oral; once daily for 14 days) improves certain autism spectrum disorder (ASD)-like symptoms in animals caught in models of chronic neurodevelopmental abnormalities [1]. TAK-418 elevates H3K4me1/2/3 and H3K9me2 levels of the Ucp2 gene and stimulates Ucp2 mRNA expression in primary cultured neurons. TAK-418 also elevates H3K4me1/2/3 of the Bdnf gene. TAK-418 avoids steric hindrance of GFI1B in the binding pocket by creating a tightly formylated adduct form of coenzyme flavin adenine dinucleate (FAD). TAK-418 demonstrates excellent pharmacokinetic characteristics in omnidids and inhibits the LSD1 enzyme in the brain. A single injection of 1 or 3 mg of TAK-418/kg enhanced H3K4me2 levels of the Ucp2 gene in the mouse brain [ 1]. TAK-418 can be active without generating hematological toxicity in hungry animals [1]. Improves neurological difficulties in terms of cells, molecules, and gene expression, and functional and functional levels of KS mouse model (Kmt2d+/βGeo mice) [2].

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In valproate (VPA)-exposed rat models of autism spectrum disorder (ASD), oral administration of TAK-418 (1 mg/kg/day for 2 weeks) significantly improves sociability deficits in both juvenile (5-week-old) and adult (15-week-old) rats, while lower doses (0.1, 0.3 mg/kg) are ineffective. A 1-week treatment also rescues sociability, but a single dose is ineffective. TAK-418 (1 mg/kg/day for 16–20 days) also ameliorates cognitive impairment in VPA rats in the novel object recognition test [1].
In poly I:C-induced maternal immune activation mouse models of ASD, TAK-418 (0.1, 0.3, 1 mg/kg/day for 2 weeks) dose-dependently rescues sociability deficits [1].
TAK-418 (1 mg/kg) inhibits LSD1 enzyme activity in the rat cortex by 95.7% at 2 hours post-dose and in mouse brain by 91.5%. It almost completely normalizes genome-wide dysregulated gene expression in the cortex of VPA rats and poly I:C mice, as shown by RNA-seq, and restores aberrant DNA methylation patterns [1].
In Kmt2d\(^{+/βGeo}\) mice (a model of Kabuki syndrome), oral TAK-418 (1 mg/kg/day for 2 weeks) rescues adult hippocampal neurogenesis deficits (increased DCX⁺ cells) in a dose-dependent manner (full effect at 1 mg/kg, partial at 0.5 and 2 mg/kg). It restores H3K4me1/2 levels and partially restores H3K4me3 in the hippocampus, and reverses abnormal gene expression (e.g., Fos, Fosb). TAK-418 also improves spatial learning and memory deficits in the Morris water maze and corrects splenomegaly after 8 weeks of treatment [2].

LSD1 enzyme kinetics are measured using a peroxidase-coupled assay. Recombinant human LSD1/CoREST complex is incubated with various concentrations of TAK-418 in assay buffer containing monomethylated H3K4 peptide. Horseradish peroxidase and Amplex Red are added, and the hydrogen peroxide produced during the amine oxidase reaction is monitored fluorometrically (excitation 570 nm, emission 585 nm). Progress curves are fitted to obtain \(k_{\text{obs}}\), and \(k_{\text{inact}}/K_{\text{I}}\) values are calculated from replots of \(k_{\text{obs}}\) versus inhibitor concentration [1].

Primary cultured rat neurons are treated with TAK-418 (0.0001–10 µM) or DMSO for 3 days. Chromatin immunoprecipitation (ChIP) is performed using antibodies against H3K4me2 and H3K9me2, followed by qPCR at the Ucp2 gene. Total RNA is extracted and Ucp2 mRNA levels are quantified by qRT-PCR [1].
For co-immunoprecipitation, TF-1a cells are treated with TAK-418 or T-711 for 6 h, then lysed. LSD1 is immunoprecipitated from chromatin fractions, and bound GFI1B is detected by Western blotting [1].

VPA rats are generated by injecting pregnant SD rats with sodium valproate (500 mg/kg i.p.) on gestational day 12.5. Poly I:C mice are generated by injecting pregnant C57BL/6J mice with poly I:C (5 mg/kg i.v.) on gestational day 15. Male offspring are used. TAK-418 is dissolved in 0.5% methylcellulose/0.5% citrate and administered orally once daily for 2 weeks (or 1 week, or single dose) at doses of 0.1, 0.3, or 1 mg/kg. Behavioral tests (three-chamber sociability, novel object recognition, Morris water maze) are performed after treatment. Tissues (cortex, hippocampus) are collected for RNA-seq, ChIP-seq, RRBS, Western blot, and immunohistochemistry [1, 2].
For Kabuki syndrome model, Kmt2d\(^{+/βGeo}\) mice on C57BL/6J background are orally gavaged daily with TAK-418 (1 mg/kg) or vehicle for 2 weeks (neurogenesis and molecular studies) or 8 weeks (splenomegaly study). Brains are processed for DCX immunofluorescence, histone extraction, ChIP-seq, RNA-seq; spleens are weighed [2].

TAK-418 exhibits favorable pharmacokinetic characteristics in rodents. After oral administration, it is rapidly absorbed and has good bioavailability. In mice, a single dose of 1 mg/kg increased the H3K4me2 level of the Ucp2 gene in the brain, indicating its brain permeability. Specific parameters such as half-life, Cmax and oral bioavailability are shown in Supplementary Figure [1].

TAK-418 was well tolerated in rodents. No significant weight loss or adverse clinical signs were observed at treatment doses (up to 1 mg/kg/day for 8 weeks) in VPA rats, poly I:C mice, or Kmt2d(^{+/βGeo}) mice. Unlike the conventional LSD1 inhibitor T-711, TAK-418 did not cause thrombocytopenia or hematologic toxicity [1, 2]. In a Kabuki syndrome mouse model, TAK-418 treatment for 8 weeks improved splenomegaly without significant side effects [2].

[1]. LSD1 enzyme inhibitor TAK-418 unlocks aberrant epigenetic machinery and improves autism symptoms in neurodevelopmental disorder models. Sci Adv. 2021;7(11):eaba1187.

[2]. Inhibition of KDM1A activity restores adult neurogenesis and improves hippocampal memory in a mouse model of Kabuki syndrome. Mol Ther Methods Clin Dev. 2021;20:779-791.

TAK-418 is a novel, orally bioavailable, LSD1 enzyme-specific inhibitor developed by Takeda Pharmaceutical Company Limited. It is designed to avoid interfering with the interaction between LSD1 and cofactors, thereby minimizing hematologic toxicity. Preclinical studies have shown that TAK-418 can restore abnormal epigenetic modifications and gene expression to normal in various neurodevelopmental disorder models (autism spectrum disorder, Kabuki syndrome), thereby improving behavior. TAK-418 has completed Phase I clinical trials in healthy volunteers (NCT03228433, NCT03501069) and has received orphan drug designation from the EMA and FDA for the treatment of Kabuki syndrome [1, 2]. TAK-418 is a selective, orally active LSD1 (KDM1A) enzyme inhibitor. TAK-418 unlocks abnormal epigenetic mechanisms and improves autism symptoms in neurodevelopmental disorder models.

C17H25CLN2O2S

356.9106

356.132

C, 57.21; H, 7.06; Cl, 9.93; N, 7.85; O, 8.97; S, 8.98

1818252-53-7

1818252-52-6; 1818252-53-7 (HCl);

118432651

White to yellow solid-liquid Mixture

3

4

6

23

410

2

Cl.S1C=C(C(NC2CCOCC2)=O)C=C1[C@@H]1C[C@H]1NCC1CC1

LEEWMGOHGNRDKC-NUNOUFIPSA-N

InChI=1S/C17H24N2O2S.ClH/c20-17(19-13-3-5-21-6-4-13)12-7-16(22-10-12)14-8-15(14)18-9-11-1-2-11/h7,10-11,13-15,18H,1-6,8-9H2,(H,19,20)1H/t14?,15-/m1./s1

5-((2R)-2-((cyclopropylmethyl)amino)cyclopropyl)-N-(tetrahydro-2H-pyran-4-yl)thiophene-3-carboxamide hydrochloride

TAK418; TAK-418 HCl; TAK-418; 1818252-53-7; LVM0PK6IHG; 5-((1R,2R)-2-(cyclopropylmethylamino)cyclopropyl)-N-(oxan-4-yl)thiophene-3-carboxamide; RefChem:187188; TAK 418

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 : ~55 mg/mL (~154.10 mM)
H2O : ≥ 16.67 mg/mL (~46.71 mM)

Solubility in Formulation 1: ≥ 5.5 mg/mL (15.41 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 55.0 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.

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Solubility in Formulation 2: ≥ 5.5 mg/mL (15.41 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 55.0 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.

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 (Please use freshly prepared in vivo formulations for optimal results.)