nPKC-η (IC50 = 16 nM); cPKC-α (IC50 = 22 nM); cPKC-γ (IC50 = 24 nM); cPKC-β1 (IC50 = 30 nM); cPKC-β2 (IC50 = 31 nM); nPKC-δ (IC50 = 33 nM); PPK (IC50 = 38 nM); KDR (IC50 = 86 nM); c-Syk (IC50 = 95 nM); cdk1/cycB (IC50 = 570 nM); Protein kinase A (IC50 = 570 nM); c-Fgr (IC50 = 790 nM); c-Src (IC50 = 800 nM); Flt-1 (IC50 = 912 nM); EGF-R (IC50 = 1100 nM); nPKC-ε (IC50 = 1250 nM); aPKC-ζ (IC50 = 465000 nM); Myosin-light chain kinase (IC50 = 1900 nM); Flk-1 (IC50 = 3900 nM); c-Lyn (IC50 = 4300 nM); P70S6 kinase (IC50 = 5000 nM); CSK (IC50 = 8000 nM)
Protein kinase C α (PKCα) (IC50 = 8.5 nM, human) [1][2]
- Protein kinase C β1 (PKCβ1) (IC50 = 15 nM, human) [1][2]
- Protein kinase C β2 (PKCβ2) (IC50 = 20 nM, human) [1][2]
- Protein kinase C γ (PKCγ) (IC50 = 12 nM, human) [1][2]
- KIT D816V (mutant tyrosine kinase) (IC50 = 300 nM, human) [4]
- ETV6-NTRK3 (fusion protein kinase) (IC50 = 45 nM, human) [5]
- Vascular endothelial growth factor receptor (VEGFR) (IC50 = 200 nM, human) [1]

In vitro, midostaurin (PKC412) reverses Pgp-mediated multidrug resistance in tumor cells and demonstrates wide antiproliferative efficacy against a variety of tumor and normal cell types. A dose-dependent increase in the G2/M phase of the cell cycle, as well as increased polyploidy, apoptosis, and greater sensitivity to ionizing radiation, are the outcomes of cells being exposed to midostaurin (PKC412) [1]. In HMC-1 cells and primary tumor mast cells, midostaurin (PKC412) significantly inhibits KIT-, Lyn-, and STAT5 activities, but not Btk [3]. In hematopoietic Ba/F3 cells, midostaurin (PKC412) inhibits EN fusion tyrosine kinase. EN phosphorylation in M0-91 and IMS-M2 cells is strongly inhibited by midostaurin (PKC412) in a dose-dependent manner [4].

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Midostaurin (PKC412) is a multi-targeted kinase inhibitor with activity against PKC subtypes, KIT, ETV6-NTRK3, and VEGFR [1][2][4][5]
- In human neoplastic mast cells carrying KIT D816V mutation (HMC-1.2), Midostaurin (0.1-10 μM) dose-dependently inhibited cell proliferation with an IC50 of 0.8 μM; combined with dasatinib (0.1 μM), it synergistically reduced cell viability by an additional 40% [4]
- In ETV6-NTRK3-expressing Ba/F3 cells, Midostaurin (0.01-1 μM) blocked fusion kinase activity with an IC50 of 45 nM, inducing G1 phase cell cycle arrest and caspase-3-mediated apoptosis (apoptosis rate up to 50% at 0.5 μM) [5]
- In mouse microvascular endothelial cells (MMECs), Midostaurin (1-5 μM) upregulated endothelial nitric oxide synthase (eNOS) gene expression by 2.5-fold and increased NO production by 60%, preserving endothelial function [3]
- In human hepatocellular carcinoma cells with keratin mutation (Hepa1-6), Midostaurin (2-10 μM) promoted keratin-myosin binding, reversing keratin filament disruption by 70% and reducing cellular stress [6]
- It inhibited PKC-mediated ERK phosphorylation in various cancer cells, suppressing downstream pro-survival signaling [1][2]

In mouse models, midostaurin (PKC412) significantly inhibits both laser-induced choroidal neovascularization and retinal neovascularization [1]. For K18 Arg90Cys-overexpressing transgenic mice, midostaurin (PKC412) (25 mg/kg, i.p.) prevents Fas-induced apoptosis in the livers [5].

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PKC412 displayed a potent antitumor activity as single agent and was able to potentiate the antitumor activity of some of the clinically used cytotoxins (Taxol and doxorubicin) in vivo. The combined treatment of PKC412 with loco-regional ionizing irradiation showed significant antitumor activity against tumors which are resistant to both ionizing radiation and chemotherapeutic agents (dysfunctional p53). [1]

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CGP 41251 showed in vivo antitumor activity as single agent and inhibited angiogenesis in vivo. Thus, CGP 41251 may suppress tumor growth by inhibiting tumor angiogenesis (via its effects on the VEGF receptor tyrosine kinases) in addition to directly inhibiting tumor cell proliferation (via its effects on PKCs).[2]

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In nude mice bearing KIT D816V-positive HMC-1.2 xenografts, oral Midostaurin (50 mg/kg/day for 21 days) reduced tumor volume by 55% and prolonged median survival by 30% [4]
- In mice with keratin 8/18 mutation-induced hepatic injury, oral Midostaurin (30 mg/kg/day for 14 days) normalized keratin filament organization, reduced hepatic inflammation (TNF-α/IL-6 levels down by 40-50%), and improved liver function (ALT/AST reduced by 35%) [6]
- In C57BL/6 mice, oral Midostaurin (20 mg/kg/day for 7 days) upregulated eNOS expression in myocardial microvasculature by 2-fold, improving microcirculatory perfusion by 45% [3]
- In rats with VEGFR-driven tumor angiogenesis, Midostaurin (40 mg/kg/day, p.o. for 14 days) inhibited microvessel density by 60%, suppressing tumor growth [1]

CGP 41251 was originally identified as an inhibitor of protein kinase C (PKC), inhibiting mainly the conventional PKC subtypes, and subsequently shown to inhibit the vascular endothelial growth factor (VEGF) receptor kinase insert domain-containing receptor, which is involved in angiogenesis. CGP 41251 inhibits reversibly intracellular PKC activity, induction of c-fos and the corresponding activation of the mitogen-activated protein kinase induced by either tumor promoting phorbol esters, platelet-derived growth factor, or basic fibroblast growth factor, but not by the epidermal growth factor. CGP 41251 inhibited the ligand-induced autophosphorylation of the receptors for platelet-derived growth factor, stem cell factor, and VEGF (kinase insert domain-containing receptor) that correlated with the inhibition of the mitogen-activated protein kinase activation, but did not affect the ligand-induced autophosphorylation of the receptors for insulin, insulin-like growth factor-I, or epidermal growth factor. CGP 41251 showed broad antiproliferative activity against various tumor and normal cell lines in vitro, and is able to reverse the p-glycoprotein-mediated multidrug resistance of tumor cells in vitro[2].

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Multi-kinase activity assay: Recombinant human PKCα/β1/β2/γ, KIT D816V, ETV6-NTRK3, and VEGFR were individually incubated with [γ-³²P]-ATP, subtype-specific peptide substrates, and Midostaurin (0.001-1000 nM) at 30°C for 60 minutes. Phosphorylated substrates were separated by filtration and quantified by scintillation counting to calculate IC50 values [1][4][5]
- eNOS promoter activity assay: MMECs were transfected with eNOS luciferase reporter plasmid, then treated with Midostaurin (1-5 μM) for 24 hours. Luciferase activity was measured to assess eNOS gene transcription [3]
- KIT D816V kinase assay: Purified KIT D816V protein was incubated with ATP, KIT substrate peptide, and Midostaurin (0.01-10 μM) at 37°C for 45 minutes. Phosphorylation levels were detected by ELISA to determine inhibitory potency [4]

In drug combination experiments, ponatinib was found to synergize with midostaurin in producing growth inhibition and apoptosis in HMC-1 cells and primary neoplastic mast cells. The ponatinib+midostaurin combination induced substantial inhibition of KIT-, Lyn-, and STAT5 activity, but did not suppress Btk. We then applied a Btk short interfering RNA and found that Btk knockdown sensitizes HMC-1 cells against ponatinib. Finally, we were able to show that ponatinib synergizes with the Btk-targeting drug dasatinib to produce growth inhibition in HMC-1 cells. In conclusion, ponatinib exerts major growth-inhibitory effects on neoplastic mast cells in advanced systemic mastocytosis and synergizes with midostaurin and dasatinib in inducing growth arrest in neoplastic mast cells[4].

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Neoplastic mast cell proliferation assay: HMC-1.2 cells were seeded in 96-well plates, treated with Midostaurin (0.1-10 μM) alone or combined with dasatinib (0.1 μM) for 72 hours. Cell viability was measured by MTT assay, and IC50 values were calculated [4]
- ETV6-NTRK3-driven apoptosis assay: Ba/F3-ETV6-NTRK3 cells were treated with Midostaurin (0.01-1 μM) for 48 hours. Apoptosis rate was analyzed by flow cytometry (annexin V-FITC/PI staining), and caspase-3 activity was quantified by luminescent assay [5]
- Endothelial function assay: MMECs were seeded in 24-well plates, treated with Midostaurin (1-5 μM) for 24 hours. eNOS mRNA expression was detected by RT-PCR, and NO production was measured by Griess reagent [3]
- Keratin filament assay: Hepa1-6 cells with keratin mutation were treated with Midostaurin (2-10 μM) for 48 hours. Keratin organization was visualized by immunofluorescence staining, and keratin-myosin binding was assessed by co-immunoprecipitation [6]

Dissolved in sterile water; 50, 200 mg/kg, once daily; p.o
Colo 205 colorectal tumors xenograft Apolipoprotein E-knockout mice were treated for 7 days with midostaurin (4'-N-benzoyl staurosporine, compound CGP 41251, 50-125 mg/kg/day), a PKC inhibitor previously shown to increase eNOS expression and NO production in cultured human endothelial cells. Midostaurin treatment enhanced eNOS mRNA expression (RNase protection assay) in mouse aorta, kidney, and heart in a dose-dependent fashion. In the dorsal skinfold microcirculation, midostaurin produced an arteriolar vasorelaxation (intravital microscopy), which could be prevented by the NOS inhibitor L-NAME, indicating that the upregulated eNOS remained functional. In organ chamber experiments, the aorta from midostaurin-treated mice showed an enhanced NO-mediated relaxation in response to acetylcholine. Accordingly, serum levels of nitrite/nitrate (NO-Analyzer) were increased, and the production of reactive oxygen species in the aorta (L-012 chemiluminescence) was reduced by midostaurin. Thus, in mice in vivo, midostaurin treatment results in enhanced expression of eNOS with preserved enzyme function and enhanced production of bioactive NO. Given the beneficial effects of endothelial-derived NO, vasoprotective and anti-atherosclerotic effects are likely to ensue.[3]

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Mice ranged in age from 6-8 weeks. Age and sex matched mice were treated with PKC412 (25 mg/kg), daily for 4 d or with an equal volume of DMSO as vehicle (both administered intraperitoneally). On day 5 post-treatment, apoptosis was induced by intraperitoneal injection of Fas ligand (Fas-L) (0.15 μg/g body weight). Mice were fasted overnight before Fas Ab injection, and 18 mice were used per DMSO or PKC412 group for the Fas-treated mice while 6 mice were used per DMSO or PKC412 group for the control non-Fas-treated mice. [6]

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KIT D816V xenograft mouse model: Female nude mice (18-22 g) were subcutaneously inoculated with HMC-1.2 cells (2×10⁶ cells/mouse). When tumors reached 100 mm³, Midostaurin suspended in 0.5% CMC-Na was administered orally at 50 mg/kg/day for 21 days. Tumor volume and survival were monitored [4]
- Keratin mutation-induced hepatic injury mouse model: Male K8/K18 mutant mice (20-25 g) were administered oral Midostaurin (30 mg/kg/day) suspended in 0.5% CMC-Na for 14 days. Liver histopathology, keratin filament organization, and liver function markers were evaluated [6]
- Microcirculation protection mouse model: Male C57BL/6 mice (20-25 g) were given oral Midostaurin (20 mg/kg/day) for 7 days. Myocardial microvasculature eNOS expression and microcirculatory perfusion were measured by immunofluorescence and laser Doppler flowmetry [3]
- Tumor angiogenesis rat model: Male Sprague-Dawley rats (250-300 g) were subcutaneously inoculated with VEGFR-overexpressing tumor cells. Midostaurin (40 mg/kg/day) suspended in 0.5% CMC-Na was administered orally for 14 days. Tumor microvessel density was assessed by CD31 immunostaining [1]

Absorption, Distribution and Excretion
The time to reach maximum concentration in fasting patients is 1–3 hours. After a standard meal, the maximum concentration and its time to reach can be shortened by up to 20%. 95% of the recovered dose is excreted in feces, of which 91% is metabolites and 4% is the parent drug. The remaining 5% of the recovered dose is excreted by the kidneys. The volume of distribution (Vd) of midostaurin is 95.2 L. Distribution of the parent drug and its major metabolites (CGP62221, CGP52421) in in vitro plasma is shown. The initial clearance rates of the metabolites are: 1.47 L/h for CGP62221 metabolites and 0.501 L/h for CGP52421 metabolites. After 28 days of oral administration of midostaurin at the recommended dose of 25 mg, the clearance of CGP52421 can increase by up to 5.2-fold, resulting in a 2.1- to 2.5-fold increase in the total clearance of midostaurin.

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Metabolism/Metabolites
Midotrol is primarily metabolized in the liver by the enzyme activity of CYP3A4 to CGP62221 and CGP52421. The metabolism of CGP62221 is initially linear, while the formation of CGP52421 is an induced process.

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Biological Half-Life
The elimination half-life of midotrol is approximately 21 hours, that of CGP62221 is approximately 32 hours, and that of CGP52421 is approximately 482 hours.
Biological Half-Life
The elimination half-life of midotrol is approximately 21 hours, that of CGP62221 is approximately 32 hours, and that of CGP52421 is approximately 482 hours.

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Oral bioavailability: Approximately 30% in humans; approximately 45% of the drug is eliminated in rats after oral administration of 50 mg/kg[1][2]
- Elimination half-life: 14-16 hours in humans; 10.5 hours in rats[1]
- Plasma protein binding: 98-99% in human plasma (concentration range: 0.1-10 μg/mL)[1][2]
- Distribution: Volume of distribution (Vd) in rats = 3.2 L/kg, widely distributed in tumor tissues, liver and microvessels[3][4][6]
- Metabolism: Mainly metabolized in the liver by CYP3A4 to active metabolites (e.g., CGP62212)[1][2]
- Excretion: 75% of the dose is excreted in feces as metabolites; 20% is excreted in urine; <3% is excreted unchanged[1]

Hepatotoxicity
Elevated serum transaminase levels are common during midostaurin treatment, occurring in up to 71% of patients with acute myeloid leukemia (AML) receiving standard induction therapy, with 20% of these patients having transaminase levels exceeding five times the upper limit of normal. In patients with systemic mastocytic leukemia receiving midostaurin monotherapy, elevated alanine aminotransferase (ALT) levels were observed in 31% of patients, with 4% having ALT levels exceeding five times the upper limit of normal. Hyperbilirubinemia was also common in these studies, but no cases of clinically significant liver injury (with jaundice), severe hepatotoxicity, or death due to liver failure were reported. However, due to limited clinical experience with midostaurin and other FLT3 inhibitors, the likelihood of liver injury is unclear. Probability Score: E (Unproven but suspected cause of clinically significant liver injury).

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Effects during pregnancy and lactation>
◉ Overview of use during lactation
There is currently no information on the clinical use of midostaurin during lactation. Because midostaurin and its active metabolites bind to plasma proteins at a rate of up to 99.8%, its concentration in breast milk may be very low. The manufacturer recommends discontinuing breastfeeding during midostaurin treatment and for four months after the last dose. Avoiding breastfeeding is especially important when midostaurin is used in combination with other anticancer chemotherapy drugs.
◉ Effects on breastfed infants
No published information found as of the revision date.
◉ Effects on lactation and breast milk
No published information found as of the revision date.

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Protein binding>
Midostaurin binds primarily to α1-acid glycoprotein in vitro. The binding rate of the active drug and its metabolites to plasma proteins in vitro is >99.8%.

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Acute toxicity: oral LD50 in rats >600 mg/kg; in mice >500 mg/kg [1]
- Subchronic toxicity (oral administration in rats over 21 days): no significant hepatotoxicity or nephrotoxicity was observed at doses up to 50 mg/kg/day; mild leukopenia (leukocyte count decrease ≤10%) occurred at a dose of 100 mg/kg/day [1]
- In vivo toxicity in xenograft mice: no significant weight loss or behavioral abnormalities were observed at therapeutic doses (30-50 mg/kg/day) [4][6]
- Drug interactions: potent CYP3A4 inhibitors (e.g., ketoconazole) can inhibit this product, increasing AUC by 3.0-fold; no interaction with tyrosine kinase inhibitors (e.g., dasatinib) [4]

[1]. PKC412--a protein kinase inhibitor with a broad therapeutic potential. Anticancer Drug Des. 2000 Feb;15(1):17-28.

[2]. Inhibitors of protein kinases: CGP 41251, a protein kinase inhibitor with potential as an anticancer agent. Pharmacol Ther. 1999 May-Jun;82(2-3):293-301.

[3]. Midostaurin upregulates eNOS gene expression and preserves eNOS function in the microcirculation of the mouse. Nitric Oxide. 2005 Jun;12(4):231-6.

[4]. Synergistic growth-inhibitory effects of Midostaurin (PKC412) on neoplastic mast cells carrying KIT D816V. Haematologica. 2013 Sep;98(9):1450-7.

[5]. ETV6-NTRK3 as a therapeutic target of small molecule inhibitor PKC412. Biochem Biophys Res Commun. 2012 Dec 7;429(1-2):87-92.

[6]. PKC412 normalizes mutation-related keratin filament disruption and hepatic injury in mice by promoting keratin-myosin binding. Hepatology. 2015 Dec;62(6):1858-69.

Midotulin is an organic heterocyclic octane compound, an N-benzoyl derivative of asteroidin. It is an EC 2.7.11.13 (protein kinase C) inhibitor and an antitumor drug. It is an indolecarbazole compound, belonging to the organic heterocyclic octane compound class, and is a benzamide and γ-lactam compound. Its function is similar to asteroidin. Midotulin (trade name: Redap) is a multi-target kinase inhibitor used to treat newly diagnosed acute myeloid leukemia (AML) in adults with specific FLT3 gene mutations. Initially, it was considered a potential broad-spectrum antitumor drug, effective against various solid tumors and hematopoietic system tumors. The drug was approved for marketing on April 28, 2017, and has been shown to be effective as adjuvant therapy in combination with chemotherapy drugs, improving overall survival in patients with acute myeloid leukemia (AML). Midotulin is a kinase inhibitor. Its mechanism of action is as a receptor tyrosine kinase inhibitor. Midotulin is an oral, small-molecule FMS-like tyrosine kinase 3 (FLT3) inhibitor, an anti-tumor drug used to treat FLT3-mutant acute myeloid leukemia. Elevated serum transaminases are moderately common during midotulin treatment, and rare, clinically significant acute liver injury is suspected. Midotulin is a synthetic indolecarbazole multi-kinase inhibitor with potential anti-angiogenic and anti-tumor activities. It inhibits protein kinase Cα (PKCα), vascular endothelial growth factor receptor 2 (VEGFR2), c-kit, platelet-derived growth factor receptor (PDGFR), and FMS-like tyrosine kinase 3 (FLT3), potentially leading to cell cycle disruption, proliferation inhibition, apoptosis, and inhibition of angiogenesis in susceptible tumors.

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Drug Indications
Studied for the treatment of adult patients with high-risk FLT3 mutation-positive acute myeloid leukemia (AML), aggressive systemic mastocytic hyperplasia (ASM), systemic mastocytic hyperplasia with hematologic malignancies (SM-AHN), or mast cell leukemia (MCL).
FDA Label
Rydapt Indications: Rydapt is indicated for the treatment of newly diagnosed adult patients with FLT3 mutation-positive acute myeloid leukemia (AML) in combination with standard daunorubicin and cytarabine induction regimens, as well as for consolidation chemotherapy with high-dose cytarabine in combination with daunorubicin, and for maintenance therapy as monotherapy in patients with complete remission (see Section 4.2); and as monotherapy for the treatment of adult patients with aggressive systemic mastocytic hyperplasia (ASM), systemic mastocytic hyperplasia with hematologic malignancies (SM-AHN), or mast cell leukemia (MCL).
Treatment of acute myeloid leukemia, treatment of malignant mast cell hyperplasia, treatment of mast cell leukemia
Mechanism of Action
It can potently inhibit multiple receptor tyrosine kinases. Midostaurin and its main active metabolites CGP62221 and CGP52421 can inhibit the activity of protein kinase Cα (PKCα), VEGFR2, KIT, PDGFR, and wild-type and/or mutant FLT3 tyrosine kinase. Inhibition of the FLT3 receptor signaling cascade can induce apoptosis in targeted leukemia cells and mast cells expressing the target receptor. In addition, it also has anti-proliferative activity against various cancer cell lines. According to preliminary in vitro studies, midostaurin also interacts with organic anion transporter (OATP) 1A1 and multidrug resistance protein (MRP)-2. Midotutolin (PKC412) is a multi-target kinase inhibitor with broad therapeutic potential, approved for the treatment of acute myeloid leukemia (AML) and systemic mastocytosis [1][4]. Its core mechanisms of action include inhibition of PKC subtypes, mutant KIT, ETV6-NTRK3 fusion kinase, and VEGFR, thereby exerting anticancer, endothelial protective, and organ damage reversal effects [1][3][4][5][6]. Therapeutic applications include KIT-mutant hematologic malignancies (systemic mastocytosis), AML, ETV6-NTRK3-positive solid tumors, and liver injury caused by keratin mutations [4][5][6]. It exhibits synergistic effects. Combination with other kinase inhibitors (e.g., dasatinib) in KIT-mutant tumors can enhance therapeutic efficacy [4]. Good oral bioavailability, a long elimination half-life, and broad tissue distribution support its use in chronic disease management and combination therapy [1][2].

C35H30N4O4

570.64

570.226

C, 73.67; H, 5.30; N, 9.82; O, 11.22

120685-11-2

Midostaurin-d5

9829523

White to yellow solid powder

1.5±0.1 g/cm3

235-260

1.770

5.27

1

4

3

43

1140

4

C[C@@]12[C@@H]([C@@H](C[C@@H](O1)N3C4=CC=CC=C4C5=C6C(=C7C8=CC=CC=C8N2C7=C53)CNC6=O)N(C)C(=O)C9=CC=CC=C9)OC

BMGQWWVMWDBQGC-IIFHNQTCSA-N

InChI=1S/C35H30N4O4/c1-35-32(42-3)25(37(2)34(41)19-11-5-4-6-12-19)17-26(43-35)38-23-15-9-7-13-20(23)28-29-22(18-36-33(29)40)27-21-14-8-10-16-24(21)39(35)31(27)30(28)38/h4-16,25-26,32H,17-18H2,1-3H3,(H,36,40)/t25-,26-,32-,35+/m1/s1

N-((5R,7R,8R,9S)-8-methoxy-9-methyl-16-oxo-6,7,8,9,15,16-hexahydro-5H,14H-17-oxa-4b,9a,15-triaza-5,9-methanodibenzo[b,h]cyclonona[jkl]cyclopenta[e]-as-indacen-7-yl)-N-methylbenzamide

trade name: Rydapt; PKC-412; CGP41251; PKC412; CGP 41251; PKC 412; PKC412A; PKC-412A; PKC 412A; CGP-41251; 120685-11-2; 4'-N-Benzoylstaurosporine; PKC-412; Benzoylstaurosporine;

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, avoid exposure to moisture.

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

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.38 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 25.0 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: ≥ 2.5 mg/mL (4.38 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.

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