Aurora B (IC50 = 10.37 nM)
Barasertib-HQPA (3 μM, 3 hours) considerably reduces the expression of histone H3 phosphorylation in newly isolated leukemia cells[1].
Barasertib-hydroxyquinazoline pyrazol anilide (HQPA)] is quickly changed in plasma to the active form of barasertib-HQPA[2].
Barasertib-HQPA is employed in the in vitro research[3].
Barasertib-HQPA causes a polyploid population to emerge along with a noticeable anti-propliferative effect that, in most cases, results in apoptosis[4].

Barasertib-HQPA (3 μM, 3 hours) considerably reduces the expression of histone H3 phosphorylation in newly isolated leukemia cells[1].
Barasertib-hydroxyquinazoline pyrazol anilide (HQPA)] is quickly changed in plasma to the active form of barasertib-HQPA[2].
Barasertib-HQPA is employed in the in vitro research[3].
Barasertib-HQPA causes a polyploid population to emerge along with a noticeable anti-propliferative effect that, in most cases, results in apoptosis[4].

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The active metabolite AZD1152-HQPA inhibited the proliferation of a panel of human leukemia cell lines (including HL-60, NB4, MOLM13, PALL-2, MV4-11, EOL-1, K562) with IC50 values ranging from 3 nM to 40 nM, as measured by [³H]-thymidine uptake after 48 hours of treatment. [1]
AZD1152-HQPA inhibited the clonogenic growth (colony formation) of MOLM13 and MV4-11 cell lines with IC50s of 1 nM and 2.8 nM, respectively. [1]
In freshly isolated primary leukemia cells from patients (n=10), AZD1152-HQPA inhibited colony formation with IC50s less than 3 nM in all cases. In contrast, the IC50 for bone marrow mononuclear cells from healthy volunteers was >10 nM. [1]
AZD1152-HQPA (10 nM, 24 hours) significantly decreased the population of MOLM13 and MV4-11 cells expressing phosphorylated histone H3 (Ser10), a substrate of Aurora B kinase, confirming target inhibition. A similar effect was observed in primary patient cells. [1]
Treatment with AZD1152-HQPA (1-10 nM) induced a dose- and time-dependent accumulation of cells with 4N/8N DNA content (polyploidy) in MOLM13 and PALL-2 cells, indicating mitotic exit without cytokinesis. [1]
AZD1152-HQPA (1-10 nM) induced apoptosis in MOLM13 cells in a dose-dependent manner, as measured by Annexin V/PI staining. For example, 3 nM and 10 nM induced 6% and 50% apoptosis, respectively, at 48 hours. [1]
AZD1152-HQPA synergistically enhanced the antiproliferative effects of vincristine (a tubulin depolymerizing agent) and daunorubicin (a topoisomerase II inhibitor) against PALL-2 and MOLM13 cells, with Combination Index (CI) values < 1. [1]
Western blot analysis showed that AZD1152-HQPA enhanced the cleavage of PARP (a marker of apoptosis) induced by vincristine or daunorubicin in PALL-2 and MOLM13 cells. [1]

Barasertib (AZD1152, 25 mg/kg) significantly inhibits the weight gain and growth of tumors treated with AZD1152[1].
Barasertib (AZD1152, 5 mg/kg) improves vincristine's or daunorubicin's capacity to stop human MOLM13 leukemic xenografts from proliferating[1].
Barasertib (AZD1152, (10-150 mg/kg/d)significantly reduced the growth of xenografts of human colon, lung, and hematologic tumors (mean tumor growth inhibition range, 55% to z100%; P < 0.05)[2].

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In an immunodeficient mouse xenograft model with subcutaneously implanted MOLM13 leukemia cells, intraperitoneal administration of AZD1152 (25 mg/kg, 4 times a week for 2 weeks) significantly inhibited tumor growth and reduced final tumor weight compared to the control group (mean tumor volume: 71 vs 1261 mm³; mean tumor weight: 78 vs 583 mg). No signs of wasting or other toxicity were observed at this dose. Histological examination of tumors from treated mice showed necrotic tissue with phagocytic cell infiltration, with no detectable leukemia cells. [1]
AZD1152 (5 mg/kg) potentiated the antitumor activity of vincristine (0.2 mg/kg) in the MOLM13 xenograft model. The combination almost completely inhibited tumor growth, resulting in significantly lower tumor weights compared to either single agent, without causing weight loss. [1]
AZD1152 (5 mg/kg) also enhanced the action of daunorubicin (1 mg/kg) in the same model, leading to greater tumor growth inhibition. However, daunorubicin treatment alone at this schedule was toxic to the mice, as indicated by body weight loss. [1]

In vitro studies. [2]
Phospho-histone H3 (PhH3) suppression was determined by high-content image analysis screening. SW620 cells, seeded in 96-well plates, were incubated with AZD1152-HQPA for 24 h before being fixed in 3.7% formaldehyde for 30 min. Cells were then washed with PBS, permeabilized with 0.5% Triton X-100, and stained with rabbit anti-PhH3 (Ser10) antibodies (1:100) for 1 h at room temperature. After washing with PBS, cells were incubated with Alexa Fluor 488 goat anti-rabbit antibodies (1:200) and Hoechst stain (1:10,000) for 1 h at room temperature. Cellular levels of PhH3 were analyzed on the Array Scan II using the Target Activation algorithm to calculate the percentage of PhH3-positive cells. Individual IC50 values were calculated in Origin (version 7.5) and the data were summarized using the geometric mean (i.e., the average of the logarithmic values converted back to a base 10 number).

Cell Proliferation Assay[1].
Cell Types: AML lines (HL-60, NB4, MOLM13), ALL line (PALL-2), biphenotypic leukemia (MV4-11), acute eosinophilic leukemia (EOL-1), and the blast crisis of chronic myeloid leukemia K562 cells.
Tested Concentrations: 0-100 nM. (Barasertib -HQPA)
Incubation Duration: 48 h.
Experimental Results: IC50 values ranged from 3 nM to 40 nM.

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Colony-forming assay[1]
The effects of AZD1152 on clonogenic growth of leukemia cells as well as normal bone marrow mononuclear cells were assessed by colony-forming assay using methylcellulose medium H4534, as previously described.

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Cell-cycle analysis by flow cytometry[1]
Cell-cycle analysis was performed on leukemia cells incubated with AZD1152-HQPA (1-10 nM) for 2 days at 5 × 105 cells/mL in 12-well plates.

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Apoptosis assays[1]
The ability of AZD1152-HQPA to induce apoptosis of leukemia cells was measured by annexin V–FITC apoptosis detection kit according to the manufacturer's instructions.

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Proliferation Assay ([³H]-Thymidine Uptake): Leukemia cells were cultured in liquid medium with various concentrations of AZD1152-HQPA (1-100 nM) for 48 hours. [³H]-thymidine was added for the final 6 hours of culture. Cells were then harvested, and incorporated radioactivity was measured to assess DNA synthesis and cell proliferation. The concentration causing 50% growth inhibition (IC50) was calculated from dose-response curves. [1]
Clonogenic (Colony-Forming) Assay: Leukemia cells (1x10⁵ cells/mL for cell lines, primary patient cells for primary samples) were mixed with methylcellulose-based semisolid medium containing cytokines. AZD1152-HQPA (0.3-10 nM) or vehicle control was added to the wells. Cells were plated and incubated for 10-14 days at 37°C in a humidified 5% CO₂ atmosphere. Colonies (clusters of >50 cells) were then counted under a microscope. The IC50 for clonogenic growth was determined. [1]
Phospho-Histone H3 (Ser10) Flow Cytometry: Cells were treated with AZD1152-HQPA (e.g., 3-10 nM) for specified times (e.g., 3 or 24 hours). Cells were then fixed, permeabilized, and stained with a fluorescently labeled antibody specific for phosphorylated histone H3 (Ser10). The percentage of positive cells was quantified using flow cytometry. [1]
Cell Cycle Analysis by Flow Cytometry: Cells were treated with AZD1152-HQPA (1-10 nM) for 24 or 48 hours. Cells were harvested, fixed, stained with a DNA-binding dye (e.g., propidium iodide), and analyzed by flow cytometry. The distribution of cells in different phases (G0/G1, S, G2/M) and with polyploid DNA content (4N, 8N) was determined. [1]
Apoptosis Assay (Annexin V/Propidium Iodide Staining): Cells were treated with AZD1152-HQPA (1-10 nM) for 24 or 48 hours. Both adherent and floating cells were collected, stained with FITC-conjugated Annexin V and propidium iodide (PI), and analyzed by flow cytometry. Cells in early apoptosis (Annexin V+/PI-), late apoptosis/necrosis (Annexin V+/PI+), and viable states (Annexin V-/PI-) were distinguished. [1]
Western Blot Analysis: Cells were treated with AZD1152-HQPA and/or chemotherapeutic agents (vincristine, daunorubicin) for specified times (e.g., 12 hours). Cells were lysed, proteins were quantified, separated by SDS-PAGE, and transferred to membranes. Membranes were probed with primary antibodies against target proteins such as cleaved PARP (to detect apoptosis) and α-tubulin (loading control), followed by appropriate secondary antibodies and detection via chemiluminescence. [1]
Combination Index Analysis: PALL-2 or MOLM13 cells were cultured with various concentrations of AZD1152-HQPA (0.3-10 nM) and vincristine (0.1-1 µM) or daunorubicin (3-30 nM) for 48 hours. Proliferation was assessed by [³H]-thymidine uptake. Dose-response data were analyzed using the median-effect method (Chou-Talalay) to calculate the Combination Index (CI) at different effect levels (e.g., IC50, IC75, IC90). A CI < 1 indicates synergy. [1]

Mice[1]
\nFemale immune-deficient BALB/c nude mice at 4 weeks of age were were maintained in pathogen-free conditions with irradiated chow. Animals were bilaterally, subcutaneously injected with 2 × 106 MOLM13 cells/tumor in 0.1 mL Matrigel or every another day, respectively. Daunorubicin (1 mg/kg) was given to mice by intraperitoneal injection 6 times during 2 weeks of treatment either alone or in combination with AZD1152 (5 mg/kg). The dose of these agents was determined by our preliminary studies (data not shown). Control diluent was given to the untreated control mice. Body weight and tumors were measured twice a week. Tumor sizes were calculated by the formula: a × b × c, where “a” is the length, “b” is the width, and “c” is the height in millimeters.

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\nIn vivo studies. Male Swiss nude (nu/nu genotype), SCID-bg mice (CB17/Icr.Cg.PrkdcSCIDLystbg/Crl), or nude rats (Nude:Hsd Han:RNU-rnu; AstraZeneca) were housed in negative pressure isolators or in an individually ventilated cage system. Experiments were conducted on 8- to 12-week-old animals. Human tumor xenografts were established by s.c. injecting 100 to 200 μL tumor cells (between 1 × 106 and 1 × 107 cells mixed 50:50 with Matrigel; Becton Dickinson) on the flank. Animals were randomized into treatment groups (n = 8-11 per group) when tumors reached a defined palpable size (0.2-0.3 cm3 and 0.5-1 cm3 for mice and rats, respectively). AZD1152 was prepared in Tris buffer (pH 9) and administered either as a bolus injection (i.v. or i.p.) or as a continuous 48-h infusion via s.c. implanted osmotic mini-pumps (two 24-h pumps implanted sequentially.) in accordance with the manufacturer's instructions. Tumors were measured up to three times weekly with calipers, tumor volumes were calculated, and the data were plotted using the geometric mean for each group versus time. Tumor volume and tumor growth inhibition were calculated as described previously. Statistical analysis of any change in tumor volume was carried out using a Student's one-tailed t test (P value of <0.05 was considered to be statistically significant).[2]
\nFor pharmacodynamic time course studies, nude rats bearing established SW620 tumor xenografts received vehicle or AZD1152 (25 mg/kg/d) as a daily i.v. bolus dose for 4 consecutive days (days 1-4). At multiple time points after dosing (days 0, 5, 9, 12, 16, and 19), two subgroups (n = 3 per group) of either vehicle- or AZD1152-treated animals were humanely killed and tumor and normal proliferating tissues (including bone marrow) were excised and assessed for pharmacodynamic effects using flow cytometric, histologic, or immunohistochemical analysis.[2]

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\nFemale immune-deficient BALB/c nude mice (MOLM13 cells injected)[1].
\n5 or 25 mg/kg.
\nIntraperitoneal injection 4 times a week or every another day.

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\nMOLM13 Xenograft Model: Female immunodeficient BALB/c nude mice (4 weeks old) were subcutaneously injected bilaterally with MOLM13 leukemia cells (2x10⁶ cells/tumor) suspended in Matrigel. When palpable tumors formed, mice were randomized into treatment groups (n=5 mice/group, 10 tumors/group). [1]
\nAZD1152 Monotherapy: AZD1152 (formulated in 3 M Tris buffer, pH 9.0, at 2.5 mg/mL) was administered via intraperitoneal (i.p.) injection at doses of 5 or 25 mg/kg, four times a week for two weeks. Control mice received the diluent. [1]
\nCombination Therapy with Vincristine: AZD1152 (5 mg/kg, i.p., 4 times/week for 2 weeks) was co-administered with vincristine (0.2 mg/kg, i.p., every other day during the first week). [1]
\nCombination Therapy with Daunorubicin: AZD1152 (5 mg/kg, i.p., 4 times/week for 2 weeks) was co-administered with daunorubicin (1 mg/kg, i.p., 6 times over 2 weeks). [1]
\nTumor dimensions (length, width, height) were measured twice weekly to calculate volume. Body weight was monitored as an indicator of toxicity. At the end of the experiment (2 weeks), mice were euthanized, tumors were excised and weighed, and tissue samples were collected for histological analysis. [1]

AZD1152 is described as a prodrug that can be rapidly converted into its active metabolite AZD1152-HQPA in human plasma. [1]

In mouse xenograft studies, mice treated with AZD1152 alone at doses of 5 or 25 mg/kg did not show emaciation (weight loss) or other significant toxicities. However, combination therapy with daunorubicin (1 mg/kg) was found to be toxic, resulting in weight loss in mice. [1]
The literature indicates that in a phase I clinical trial (not within the scope of the results of this preclinical study), the dose-limiting toxicity of AZD1152 was neutropenia. [1]

[1]. AZD1152, a novel and selective aurora B kinase inhibitor, induces growth arrest, apoptosis, and sensitization for tubulin depolymerizing agent or topoisomerase II inhibitor in human acute leukemia cells in vitro and in vivo.Blood. 2007 Sep 15;110(6):2034-40.

[2]. AZD1152, a selective inhibitor of Aurora B kinase, inhibits human tumor xenograft growth by inducing apoptosis. Clin Cancer Res. 2007 Jun 15;13(12):3682-8.

[3]. The selective Aurora B kinase inhibitor AZD1152 is a potential new treatment for multiple myeloma. Br J Haematol. 2008 Feb;140(3):295-302.

[4]. AZD1152 rapidly and negatively affects the growth and survival of human acute myeloid leukemia cells in vitro and in vivo. Cancer Res. 2009 May 15;69(10):4150-8.

AZT-1152 is a dihydrophosphate prodrug whose active form is the pyrazoloquinazoline Aurora kinase inhibitor AZD1152-hydroxyquinazoline pyrazolaniline (HQPA), which is rapidly converted to the active drug AZD1152-HQPA in plasma. It possesses multiple pharmacological effects, including as a prodrug, antitumor drug, and Aurora kinase inhibitor. AZT-1152 belongs to the quinazoline, monoalkyl phosphate, aniline, monofluorobenzene, pyrazole, secondary amino compounds, secondary carboxamides, and tertiary amino compounds. It is functionally related to AZD-1152. Balazertinib has been used in clinical trials for the treatment of various diseases, including oncology, lymphoma, solid tumors, and myeloid leukemia. Balazineib is a small-molecule dihydrophosphate prodrug with high oral bioavailability. Its active ingredient is AZD1152-hydroxyquinazoline pyrazole aniline (AZD1152-HQPA), an Aurora kinase inhibitor with potential antitumor activity. After administration, AZD1152-HQPA is rapidly converted into the active drug in plasma, specifically binding to and inhibiting Aurora kinase B, leading to spindle checkpoint dysfunction and chromosome alignment disorder, thereby interfering with chromosome segregation and cytokinesis. Therefore, in tumor cells overexpressing Aurora kinase B, cell division and proliferation are inhibited, and apoptosis is induced. Aurora kinase B is a serine/threonine protein kinase whose function is to connect the mitotic spindle to the centromere; it is overexpressed in various cancer cell types. AZD-1152 is a quinazoline compound with the chemical name 4-aminoquinazoline-7-ol, in which the 4-amino group is replaced by a 5-[2-(3-fluoroaniline)-2-oxoethyl]-1H-pyrazol-3-yl group, and the 7-hydroxyl group is converted to the corresponding 3-[ethyl(2-hydroxyethyl)aminopropyl] ether. It is an antitumor drug and an Aurora kinase inhibitor. It belongs to the quinazoline class, secondary amide class, tertiary amine class, secondary amine class, pyrazol class, primary alcohol class, monofluorobenzene class, and aniline class. Defosbarasertib is a small molecule inhibitor of the serine/threonine kinase Aurora B with potential antitumor activity. After administration, defosbarasertib specifically binds to and inhibits Aurora kinase B, thereby disrupting spindle checkpoint function and chromosome alignment, leading to chromosome segregation and cytokinesis arrest. This inhibited cell division and proliferation, and induced apoptosis in tumor cells overexpressing Aurora kinase B. Aurora kinase B is a serine/threonine protein kinase that functions to link the spindle to the centromere and is overexpressed in various cancer cell types. Aurora kinases play a crucial role in chromosome alignment, segregation, and cytokinesis during mitosis. Our recent studies have shown that hematopoietic malignancies, including acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL), abnormally express Aurora A and B kinases, and that the potent inhibitor of Aurora kinase, ZM447439, effectively induced growth arrest and apoptosis in various leukemia cell lines. This study investigated the effects of the highly selective inhibitor of Aurora B kinase, AZD1152, on various human leukemia cell lines. AZD1152 inhibited the proliferation of acute myeloid leukemia (AML) cell lines (HL-60, NB4, MOLM13), acute lymphoblastic leukemia (ALL) cell line (PALL-2), biphenotypic leukemia (MV4-11), acute eosinophilic leukemia (EOL-1), and chronic myeloid leukemia (CML) K562 cells during the blast crisis, with IC50 values ranging from 3 nM to 40 nM, determined by thymidine uptake on day 2 of culture. These cells had a DNA content of 4N/8N and subsequently underwent apoptosis, as detected by cell cycle analysis and annexin V staining. Notably, AZD1152 synergistically enhanced the antiproliferative activity against MOLM13 and PALL-2 cells in vitro with vincristine (a microtubule depolymerizer) and daunorubicin (a topoisomerase II inhibitor). Furthermore, AZD1152 enhanced the effects of vincristine and daunorubicin in a MOLM13 mouse xenograft model. In summary, AZD1152 is a promising new drug for the treatment of leukemia. Further research is warranted on the combination of AZD1152 with traditional chemotherapy drugs for the treatment of leukemia patients. [1]

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Objective: This study aimed to investigate the in vivo effects of AZD1152 (a novel specific Aurora kinase inhibitor selective for Aurora B). Experimental design: The pharmacodynamic effects and efficacy of AZD1152 were determined in various human tumor xenograft models. AZD1152 was administered via various parenteral routes (subcutaneous osmotic pump, intraperitoneal injection and intravenous injection). Results: AZD1152 effectively inhibited the growth of human colon cancer, lung cancer and hematological malignancies xenografts in immunodeficient mice (mean tumor growth inhibition rate ranged from 55% to ≥100%; P<0.05). After intravenous treatment with AZD1152 in athymic rats carrying SW620 colorectal cancer tumors, detailed pharmacodynamic analysis revealed the time sequence of tumor phenotypic events: first, transient inhibition of histone H3 phosphorylation, followed by accumulation of intracellular 4N DNA (2.4 times higher than the control group), and then an increase in the proportion of polyploid cells (>4N DNA, 2.3 times higher than the control group). Histological analysis showed abnormal cell division and increased apoptosis in the tumors treated with AZD1152. Bone marrow analysis showed that the drug caused transient myelosuppression, which was completely reversible after discontinuation of AZD1152 treatment. Conclusion: These data suggest that selective targeting of Aurora B kinase may be a promising approach for the treatment of a variety of malignancies. In addition to inhibition of histone H3 phosphorylation, the detection of tumor cell polyploidy and apoptosis may also be useful biomarkers for such therapeutics. AZD1152 is currently undergoing a phase I clinical trial. [2]

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Prostate cancer is the most common non-skin cancer in men and has a high mortality rate. Prostate tumor cells exhibit different androgen receptor states. Androgen receptors play a crucial role in the progression and treatment of prostate cancer. Aurora B kinase possesses oncogenic potential and participates in chromosome segregation and cytokinesis; inhibition of Aurora B kinase is a promising anticancer therapy. This study aimed to investigate the effects of the Aurora B inhibitor AZD1152-HQPA on the survival and proliferation of androgen receptor (AR)-positive prostate cancer cells. We used LNCaP as the androgen-dependent prostate cancer cell line. We investigated the effects of AZD1152-HQPA on cell viability, DNA content, micronucleus formation, and the expression of apoptosis and cell cycle-related genes. Furthermore, we examined the expression of Aurora B and AR in 23 benign prostatic hyperplasia (BPH) specimens and 38 prostate cancer specimens. AZD1152-HQPA treatment induced LNCaP cell survival defects, polyploidization, and cell death. Fluorescence in situ hybridization (FISH) centromere labeling showed that the deletion of the entire chromosome was the source of micronuclei, indicating that AZD1152-HQPA had an aneuploidy-inducing effect. AZD1152-HQPA treatment reduced AR expression. In addition, we found that the expression of Aurora B and AR in benign prostatic hyperplasia and prostate cancer specimens was weakly positively correlated (r = 0.25, r = 0.41). This is the first time that AZD1152-HQPA can be used as an effective treatment strategy for androgen-dependent prostate cancer cell lines. AZD1152-HQPA can induce aneuploidy in micronuclei. In summary, this study provides a new idea for overcoming the obstacles in prostate cancer treatment. [3]

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Aurora kinase plays a key role in regulating mitosis and cell division, and its overexpression is associated with the survival and proliferation of human cancer cells. This study reports the in vitro and in vivo activities of compound AZD1152 (a compound selective for Aurora B kinase) in acute myeloid leukemia (AML) cell lines, primary AML samples, and cord blood cells. AZD1152 exhibited antiproliferative or cytotoxic effects in all studied cell lines, inhibiting phosphorylation of histone H3 Ser10 (pHis H3) in a dose-dependent manner and resulting in cellular DNA content >4N. In contrast to the apoptotic response observed in other cell lines, AZD1152-treated THP-1 cells accumulated in a polyploid state and exhibited a senescent response. Accordingly, AZD1152 significantly affected the growth of AML cell lines and primary AML in an in vivo xenograft model. However, concentration-dependent effects on cell growth, apoptosis, and cell cycle progression were also observed in in vitro and in vivo analyses of human cord blood and primary lineage-negative stem cells and progenitor cells. These data suggest that inhibiting Aurora B kinase may be an effective treatment strategy for acute myeloid leukemia (AML), and further exploration of dosage and treatment regimens is necessary in clinical trials. [4] AZD1152 is a novel acetanilide-substituted pyrazole-aminoquinazoline prodrug. Its active form, AZD1152-HQPA, is a highly potent and selective Aurora B kinase inhibitor, which is about 100 times more potent in leukemia cells than another Aurora kinase inhibitor, ZM447439. [1] Its mechanism of action involves the inhibition of Aurora B kinase, leading to cytokinesis failure, polyploidization (4N/8N DNA content), and subsequent apoptosis. [1] Both in vitro and in vivo experiments have shown that AZD1152 has synergistic antitumor activity with traditional chemotherapeutic drugs such as vincristine and daunorubicin. [1]
Studies have shown that AZD1152 may be a promising treatment for leukemia, including in cases resistant to imatinib, and its efficacy in combination with other chemotherapy drugs warrants further investigation. [1]

C26H31FN7O6P

587.54

587.205

C, 53.15; H, 5.32; F, 3.23; N, 16.69; O, 16.34; P, 5.2 7

722543-31-9

Barasertib-HQPA;722544-51-6; 722543-31-9 (free acid); 722543-50-2 (2HCl); 957104-91-5

11497983

Off-white to light yellow solid powder

1.5±0.1 g/cm3

1.675

1.71

5

12

15

41

859

0

O=C(NC1=CC=CC(F)=C1)CC2=CC(NC3=C4C=CC(OCCCN(CC)CCOP(O)(O)=O)=CC4=NC=N3)=NN2

GBJVVSCPOBPEIT-UHFFFAOYSA-N

InChI=1S/C26H31FN7O6P/c1-2-34(10-12-40-41(36,37)38)9-4-11-39-21-7-8-22-23(16-21)28-17-29-26(22)31-24-14-20(32-33-24)15-25(35)30-19-6-3-5-18(27)13-19/h3,5-8,13-14,16-17H,2,4,9-12,15H2,1H3,(H,30,35)(H2,36,37,38)(H2,28,29,31,32,33)

2-[ethyl-[3-[4-[[5-[2-(3-fluoroanilino)-2-oxoethyl]-1H-pyrazol-3-yl]amino]quinazolin-7-yl]oxypropyl]amino]ethyl dihydrogen phosphate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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 (4.26 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
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 (4.26 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
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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Solubility in Formulation 3: ≥ 2.17 mg/mL (3.69 mM) 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 21.7 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 4: ≥ 2.17 mg/mL (3.69 mM) 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 21.7 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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Solubility in Formulation 5: ≥ 2.17 mg/mL (3.69 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 21.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 6: 2% DMSO+40% PEG 300+2% Tween 80+ddH2O: 7mg/mL

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