| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
AKT and NQO1. KP372-1 is an inhibitor of AKT kinase activity. It has an IC50 of 250 nM for AKT inhibition in cellular assays and an IC50 of 30-60 nM for inhibiting the growth of thyroid cancer cells. Concurrently, it acts as a redox-cycling agent for NQO1, an enzyme often overexpressed in cancer cells, leading to the generation of reactive oxygen species (ROS) and subsequent DNA damage.
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| ln Vitro |
With an IC50 of 200 and 100 nM, respectively, KP372-1 (0.0625, 0.125, 0.25, 0.5, 1.0 μM; 48 hours) suppresses the development of JMARc42 and Tu167c2 cells [1]. In Tu167c2 cells, KP372-1 (125 nM; 24 h) causes apoptosis, but in JMARc42 cells, it causes anoikis [1]. In Tu167 and JMAR cells, KP372-1 (125 nM; 30 minutes) decreases phosphorylation of S6 ribosomal protein via blocking Akt [1]. KP372-1 (IC50 = 250 nM) suppresses Akt kinase activity in JMAR cells at 0.250, 0.5, and 1.0 μM for 30 minutes [1].
In vitro, KP372-1 demonstrates potent anti-proliferative activity. At low micromolar concentrations (0.25-1.0 uM), it effectively inhibits AKT kinase activity, suppresses cell proliferation, and induces apoptosis (programmed cell death) and anoikis (a form of cell death induced by loss of cell adhesion) in various cancer cell lines. For instance, in thyroid cancer cells, KP372-1 shows an IC50 in the range of 30-60 nM for inhibiting cell growth. Its mechanism also involves causing DNA fragmentation through the generation of ROS, which contributes to its potent cytotoxic effects. |
| ln Vivo |
KP372-1 (10, 20 mg/kg; intravenous injection; once daily for 33 days) inhibits the growth of tumors in vivo and causes NADH oxidation without appearing to be harmful[2].
In vivo, KP372-1 has been shown to effectively reduce tumor growth in animal models. Following systemic administration, the compound leverages the high expression of NQO1 in tumors to generate damaging ROS, while simultaneously shutting down the pro-survival AKT pathway. This combination leads to significant inhibition of cancer/tumor cell growth and the induction of cell death, leading to reduced tumor burden. |
| Enzyme Assay |
Standard cell‑free assays for KP372-1 are performed to measure its two primary targets. For AKT inhibition, a kinase activity assay is used. Recombinant active AKT1 enzyme (10-50 ng) is incubated with varying concentrations of KP372-1 (0-1000 nM) in a reaction buffer (50 mM HEPES, pH 7.5, 10 mM MgCl2, 1 mM DTT) along with a synthetic peptide substrate and 10 uM ATP. After 30 minutes at 22degC, the reaction is stopped and the level of phosphorylated substrate is measured, for example using a luminescence-based assay (e.g., ADP-Glo) or by a time-resolved FRET (TR-FRET) method. The IC50 for AKT inhibition is calculated from the resulting inhibition curve. For NQO1 redox cycling activity, a cell‑free enzymatic assay is used. Purified human NQO1 enzyme (50 nM) is incubated with varying concentrations of KP372-1 (0-100 uM) in a Tris-HCl buffer (pH 7.4) containing 100 uM NADH as the electron donor and cytochrome c (50 uM) as an electron acceptor. The reduction of cytochrome c is monitored by measuring the increase in absorbance at 550 nm over 5-10 minutes. An increase in the rate of reduction in the presence of KP372-1 indicates NQO1-dependent redox cycling activity.
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| Cell Assay |
Cell Proliferation Assay[1]
Cell Types: JMARc42 and Tu167c2 cells Tested Concentrations: 0.0625, 0.125, 0.25, 0.5, 1.0 µM Incubation Duration: 48 h Experimental Results: demonstrated antiproliferative activity. Apoptosis Analysis[1] Cell Types: Tu167c2 and JMARc42 cells Tested Concentrations: 125 nM Incubation Duration: 24 h Experimental Results: Induced approximately 90% of cells apoptosis. Western Blot Analysis[1] Cell Types: Tu167 and JMAR cells Tested Concentrations: 125 nM Incubation Duration: 30 min Experimental Results: Induced a small but consistent decrease in Akt phosphorylation with a concomitant marked decrease in S6 phosphorylation. Inhibited the EGF induced phosphorylation of Aktser473 in Tu167 and AktThr308 in JMAR. Western Blot Analysis[1] Cell Types: JMAR cells Tested Concentrations: 0.250, 0.5, 1.0 µM Incubation Duration: 30 min Experimental Results: Dramatically blocked Akt kinase activity in a dose-dependent fashion, with an IC50 of 250 nM. For in vitro cellular assays, human cancer cell lines with high NQO1 expression, such as H1299 lung carcinoma or MIA PaCa-2 pancreatic cancer cells, are seeded in 96‑well plates (5,000 cells/well) in DMEM/10% FBS. After 24 hours, the cells are treated with varying concentrations of KP372-1 (0.01-10 uM). After 48-72 hours, cell viability is measured using a luminescent CellTiter-Glo assay. To assess apoptosis, cells treated with KP372-1 for 24 hours are harvested and stained with Annexin V-FITC and propidium iodide (PI) followed by analysis via flow cytometry. For intracellular ROS measurement, cells are loaded with 10 uM DCFH-DA for 30 minutes, washed, and then treated with KP372-1. The increase in fluorescence (ex 485 nm, em 535 nm) is measured in real-time to quantify the generation of ROS. |
| Animal Protocol |
Animal/Disease Models: Nude mice (H1299 xenografts model)[2].
Doses: 10, 20 mg/kg Route of Administration: Tailvein injection; single daily for 33 days Experimental Results: Affected tumor metabolism and suppressed tumor growth. In vivo efficacy studies for KP372-1 are typically performed in athymic nude mice bearing human tumor xenografts. Female mice (6-8 weeks old) are injected subcutaneously with 5×10⁶ cancer cells (e.g., H1299). When tumors reach an average volume of 100-150 mm3, the mice are randomized into treatment groups (n=8). KP372-1 is formulated in a vehicle such as a 1:1:18 ratio of ethanol, Cremophor EL, and saline, and is administered intravenously (e.g., 2 mg/kg) or intraperitoneally (e.g., 5-10 mg/kg) daily for 2-3 weeks. Tumor volume (length × width2 × 0.5) and body weight are measured every 2-3 days. At the end of the study, tumors are excised, weighed, and processed for histology (e.g., H&E and TUNEL staining) to evaluate cell death, and lysates are prepared for Western blot analysis to assess the inhibition of AKT signaling and the induction of apoptosis markers. |
| ADME/Pharmacokinetics |
KP372-1 is a small molecule that can be formulated for in vivo administration via intravenous or intraperitoneal routes. Its pharmacokinetic properties are not extensively detailed in the provided search results. It is soluble in DMSO at concentrations up to 100 mM. Preclinical PK parameters would need to be determined for specific formulations. Given that it is a redox-cycling agent, its half-life may be relatively short.
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| Toxicity/Toxicokinetics |
Preclinical toxicity studies for KP372-1 are limited, as it is primarily a research tool. At the effective doses used in xenograft models, significant off-target toxicity was not reported, but careful monitoring of animal weight and behavior is standard. As an NQO1-targeting agent, its toxicity is expected to be higher in tissues with elevated NQO1 expression, such as certain tumor types. Standard safety precautions for handling research chemicals should be followed.
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| References |
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| Additional Infomation |
KP372-1 is a multi-targeted anticancer agent that inhibits AKT and acts as an NQO1 redox cycler. Its dual mechanism leads to potent suppression of tumor cell growth and induction of apoptosis through ROS generation. It has been evaluated in preclinical studies, but has not entered clinical trials and is not FDA-approved. It is used for research purposes. CAS: 1374996-60-7.
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| Molecular Formula |
C20H8N12O2
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| Molecular Weight |
448.36
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| Exact Mass |
448.089
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| CAS # |
1374996-60-7
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| Related CAS # |
PDK1/Akt/Flt dual pathway inhibitor;331253-86-2
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| PubChem CID |
49835991
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| Appearance |
Light yellow to yellow solid powder
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
12
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
34
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| Complexity |
699
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC=C2C(=C1)C3=NC4=NN=NN4N=C3C2=O.C1=CC=C2C(=C1)C3=NN4C(=NN=N4)N=C3C2=O
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| InChi Key |
CWFOAASSUQIXOW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/2C10H4N6O/c17-9-6-4-2-1-3-5(6)7-8(9)11-10-12-14-15-16(10)13-7;17-9-6-4-2-1-3-5(6)7-8(9)13-16-10(11-7)12-14-15-16/h2*1-4H
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| Chemical Name |
10,11,12,13,14,16-hexazatetracyclo[7.7.0.02,7.011,15]hexadeca-1(16),2,4,6,9,12,14-heptaen-8-one;10,12,13,14,15,16-hexazatetracyclo[7.7.0.02,7.011,15]hexadeca-1(16),2,4,6,9,11,13-heptaen-8-one
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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: 17.86 mg/mL (39.83 mM)
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| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2304 mL | 11.1518 mL | 22.3035 mL | |
| 5 mM | 0.4461 mL | 2.2304 mL | 4.4607 mL | |
| 10 mM | 0.2230 mL | 1.1152 mL | 2.2304 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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