| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
| Targets |
CBP (CREB-binding protein) - No specific IC50/Ki provided; TTK21 activated CBP HAT activity in a concentration-dependent manner with maximal activation at 250-275 μM in filter-binding assays. [2]
p300 - No specific IC50/Ki provided; TTK21 activated p300 HAT activity similarly at 275 μM. [2] TTK21 targets the histone acetyltransferases CBP (CREB-binding protein) and p300, which are transcriptional coactivators that regulate gene expression through histone acetylation. By activating CBP/p300, TTK21 increases histone acetylation (H2B, H3, H4K12) in the brain. This leads to the upregulation of genes involved in neurogenesis, synaptic plasticity, and memory formation. |
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| ln Vitro |
In vitro, TTK21 (100-500 μM) induced auto-acetylation of full-length recombinant p300 in a concentration-dependent manner, with significant induction at 100 μM compared to DMSO control; higher concentrations (up to 500 μM) did not further increase auto-acetylation. [2]
TTK21 alone (50-275 μM) did not induce histone acetylation in HeLa cells, as shown by Western blot analysis using anti-acetylated H3K14 antibody after 24 h treatment, indicating poor cell permeability. Sodium butyrate (5 mM, positive control) induced hyperacetylation. [2] In vitro, TTK21 activates CBP/p300 acetyltransferase activity. It increases histone acetylation in neuronal cultures. CSP-TTK21 has been shown to promote axon growth, sprouting, and synaptic plasticity in chronic experimental spinal cord injury. It also ameliorates amyloid-beta-induced deficits in plasticity and associativity in hippocampal CA1 pyramidal neurons. |
| ln Vivo |
In vivo, CSP-TTK21, upon intraperitoneal administration to mice, acetylates histones in the hippocampus and frontal cortex and increases BDNF levels. It reinstated plasticity and memory in a tauopathy mouse model, ameliorated Aβ-impaired long-term potentiation, and promoted axon growth, sprouting, and synaptic plasticity in chronic experimental spinal cord injury.
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| Enzyme Assay |
Auto-acetylation reactions of full-length p300 were performed in lysine acetyltransferase assay buffer at 30°C for 10 min with or without TTK21, followed by the addition of 1 μl of 4.7 Ci/mmol [3H]acetyl-CoA (NEN–PerkinElmer). The reaction mixture was further incubated for another 10 min at 30°C. The 3H-labeled acetylated p300 was visualized by fluorography followed by autoradiography.[1]
Filter-binding HAT assay: Highly purified HeLa core histones were incubated in HAT assay buffer at 30°C for 10 min with or without baculovirus-expressed recombinant p300 or CBP in the presence or absence of TTK21 at various concentrations (including 25, 50, 100, 200, 250, 275, 500 μM), followed by addition of 1 μl of 3.6 Ci/mmol [3H]acetyl-CoA and incubation for 10 min at 30°C. The mixture was blotted onto P-81 filter paper and radioactive counts were recorded on a liquid scintillation counter. [2] Gel fluorography HAT assay: Histones were isolated by TCA precipitation using 25% TCA, washed with acetone, dissolved in 2× SDS loading dye, heated for 5 min, separated using 15% SDS-PAGE. Coomassie staining confirmed equal loading. Gel was dehydrated in DMSO for 1 h, then incubated in scintillation fluid (2,5 diphenyl oxazole in DMSO) for 45 min, rehydrated in distilled water for 4 h, dried, and exposed to X-ray film for 5 days at -80°C. [2] p300 auto-acetylation assay: Auto-acetylation reactions of full-length p300 were performed in lysine acetyltransferase assay buffer at 30°C for 10 min with or without TTK21 (100, 200, 275, 500 μM), followed by addition of 1 μl of 4.7 Ci/mmol [3H]acetyl-CoA and further incubation for 10 min at 30°C. The 3H-labeled acetylated p300 was visualized by fluorography followed by autoradiography. [2] In vitro CBP/p300 acetyltransferase assays for TTK21 involve measuring the transfer of an acetyl group from acetyl-CoA to a histone peptide substrate. The enzyme is incubated with the substrate, acetyl-CoA, and varying concentrations of TTK21. The acetylation reaction is detected using a radioactive (³H-acetyl-CoA) or luminescence-based assay. EC₅₀ values are calculated from dose-response curves. |
| Cell Assay |
One equivalent of SOCl2 diluted in DCM was added dropwise to a suspension of 100 mg of CSP in DCM, followed by the addition of few drops of DMF . The reaction mixture was stirred at room temperature for 8–9 h. TTK21 dissolved in DCM was added dropwise to this solution. The reaction mixture was stirred for 8–9 h at room temperature. The solvent was then evaporated and washed with cold water. The crude product was centrifuged and the supernatant (i.e., water) was removed; this procedure was repeated 7–8 times. Washing was then performed using DCM and the supernatant was subsequently tested for absence of TTK21. The CSP-TTK21 conjugated was then dried at 60°C for 2–3 d.[1]
Highly purified HeLa core histones were incubated in HAT assay buffer at 30°C for 10 min with or without baculovirus-expressed recombinant p300 or CBP in the presence or absence of TTK21, followed by addition of 1 μl of 3.6 Ci/mmol [3H]acetyl-CoA (NEN–PerkinElmer) and incubated for 10 min in a final volume of 30 μl at 30°C.[1] HeLa cells were treated with TTK21 alone (50, 100, 200, 275, 500 μM) or DMSO vehicle control for 24 h. After treatment, cells were lysed, and histone acetylation was analyzed by Western blot using antibody against acetylated H3K14. No significant alteration of histone H3 acetylation was observed in response to TTK21 treatment, whereas sodium butyrate (5 mM) as positive control induced hyperacetylation. This indicates TTK21 alone is impermeable to mammalian cells. [2] In vitro cell-based assays for TTK21 are performed using neuronal cell lines or primary neurons. Cells are cultured in appropriate media and treated with TTK21 or CSP-TTK21 at various concentrations. Histone acetylation is assessed by Western blot using antibodies against acetylated histones (e.g., H2B, H3, H4K12). Gene expression changes are analyzed by RT-PCR. Neuroprotective effects are assessed in models of oxidative stress or excitotoxicity. |
| Animal Protocol |
In vivo animal studies for TTK21 are conducted in mouse models of tauopathy (e.g., P301S mice), Alzheimer's disease (Aβ-infused mice), and spinal cord injury. CSP-TTK21 is administered intraperitoneally. Cognitive function is assessed by behavioral tests (e.g., novel object recognition, Morris water maze). Synaptic plasticity is assessed by electrophysiology. Axon growth and sprouting are assessed histologically.
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| ADME/Pharmacokinetics |
Specific pharmacokinetic properties of TTK21 are characterized by its conjugation to glucose-based carbon nanospheres (CSP). CSP-TTK21 is blood-brain barrier permeable and reaches different parts of the brain. It is non-toxic and is typically administered intraperitoneally. The compound is soluble in DMSO (50 mg/ml).
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| Toxicity/Toxicokinetics |
TTK21, when conjugated to CSP, is non-toxic. In mouse studies, no toxicity was reported at the doses used. As a research compound, it is intended for laboratory use only and is not for human therapeutic use. Standard safety precautions should be followed when handling this compound.
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| References |
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| Additional Infomation |
Ttk21 is a secondary amide formed by the condensation of the carboxyl group of 2-propoxybenzoic acid with the amino group of 4-chloro-3-(trifluoromethyl)aniline. It is an activator of histone acetyltransferase CBP/p300, capable of crossing the blood-brain barrier, non-toxic, and reaching different brain regions after binding to glycosyl carbon nanospheres. It functions as a nootropic agent and a histone acetyltransferase activator. It is a secondary amide belonging to the benzamide, aromatic ether, monochlorobenzene, and (trifluoromethyl)benzene classes.
TTK21 is a small-molecule activator of CBP/p300 histone acetyltransferases, derived from the first HAT activator CTPB. It was synthesized from salicylic acid. TTK21 is poorly permeable to living cells and cannot cross the blood-brain barrier by itself. To enable cellular and in vivo activity, TTK21 was covalently conjugated to glucose-derived carbon nanospheres (CSP) using functional groups on the CSP surface. The conjugation was confirmed by infrared spectroscopy and energy-dispersive X-ray spectroscopy (detection of fluorine from the CF3 group of TTK21). CSP-TTK21 retains spherical shape and can enter mammalian cells, including neural cells and cross the blood-brain barrier after intraperitoneal administration in mice. [2] TTK21 is an activator of CBP/p300 histone acetyltransferases. When conjugated to glucose-based carbon nanospheres (CSP), it is BBB permeable and non-toxic. CSP-TTK21 promotes neurogenesis, enhances memory, and has beneficial effects in models of neurodegeneration and spinal cord injury. This product is for research use only. |
| Molecular Formula |
C17H15CLF3NO2
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|---|---|
| Molecular Weight |
357.7572
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| Exact Mass |
357.074
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| Elemental Analysis |
C, 57.07; H, 4.23; Cl, 9.91; F, 15.93; N, 3.92; O, 8.94
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| CAS # |
709676-56-2
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| PubChem CID |
68453302
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| Appearance |
White to off-white solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
369.8±42.0 °C at 760 mmHg
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| Flash Point |
177.4±27.9 °C
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| Vapour Pressure |
0.0±0.8 mmHg at 25°C
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| Index of Refraction |
1.553
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| LogP |
5.7
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
24
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| Complexity |
419
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(NC1=CC=C(Cl)C(C(F)(F)F)=C1)(=O)C1=CC=CC=C1OCCC
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| InChi Key |
AQJBXYBDNZHZRE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H15ClF3NO2/c1-2-9-24-15-6-4-3-5-12(15)16(23)22-11-7-8-14(18)13(10-11)17(19,20)21/h3-8,10H,2,9H2,1H3,(H,22,23)
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| Chemical Name |
N-[4-chloro-3-(trifluoromethyl)phenyl]-2-propoxybenzamide
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| Synonyms |
TTK-21; TTK21; TTK 21;
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~279.52 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.99 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7952 mL | 13.9758 mL | 27.9517 mL | |
| 5 mM | 0.5590 mL | 2.7952 mL | 5.5903 mL | |
| 10 mM | 0.2795 mL | 1.3976 mL | 2.7952 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.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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