AIM-100 is a potent inhibitor of Ack1 (also known as TNK2), a non-receptor tyrosine kinase. The half maximal inhibitory concentration (IC₅₀) of AIM-100 for Ack1 is 22 nmol/L. It does not inhibit the other 30 kinases tested, including PI3-kinase subfamily members and the three known AKT isoforms. [1]

AIM-100 (2-10 μM; 48 hours) causes cell cycle arrest in the G1 phase by blocking both AKT tyrosine phosphorylation and Ack1 activation. AIM-100 suppresses source Ack1 activation in addition to Ack1/AKT Tyr phosphorylation. AIM-100 suppresses AR Tyr267 phosphorylation and its instability leading to ataxia-telangiectasia (ATM) in addition to suppressing Ack1 activation. Androgen receptor (AR) binding to PSA, NKX3.1, and TMPRSS2 promoters, as well as AR polymerization activity, can all be inhibited by AIM-100 [3]. It can also prevent pTyr267-AR phosphorylation.

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In CD18 human pancreatic cancer cells, treatment with insulin led to tyrosine phosphorylation of insulin receptor and Tyr284-phosphorylation of Ack1, which was abrogated by AIM-100 treatment, leading to a concomitant decrease in AKT Tyr176-phosphorylation. [1]
In Panc-1 pancreatic cancer cells, EGF treatment induced Ack1 activation, and AIM-100 treatment inhibited both Ack1 Tyr284-phosphorylation and AKT Tyr176-phosphorylation. [1]
In MCF-7 breast cancer cells, H292 lung cancer cells, and A2780-CP ovarian cancer cells, insulin or EGF stimulation elevated pTyr284-Ack1 and pTyr176-AKT expression. AIM-100 treatment inhibited these phosphorylations and also suppressed AKT activation as indicated by decreased Ser473- and Thr308-phosphorylations of AKT. [1]
AIM-100 treatment (2-10 μmol/L for 48 hours) significantly decreased cell proliferation in pancreatic (CD18, Panc-1), ovarian (OV90), and breast (MCF-7, MDA-MB-468) cancer cell lines in MTT assays, with GI₅₀ values of approximately 7-8 μmol/L. Normal human pancreatic ductal epithelial (HPNE) cells and mouse embryonic fibroblast (MEF) cells were less sensitive, with GI₅₀ values of approximately 14-15 μmol/L. [1]
Treatment of CD18 cells with Ack1 siRNA significantly decreased cell proliferation compared to control siRNA, as shown by cell counting and WST-1 assay. [1]
Treatment of Panc-1 cells with AIM-100 (6 μmol/L for 48 hours) induced cell cycle arrest in the G1 phase, increasing the percentage of cells in G1 by 13% and decreasing cells in S phase by 10%, as determined by flow cytometry. [1]
Treatment of Panc-1 cells with AIM-100 (10 μmol/L for 48 hours) induced apoptosis, as evidenced by increased caspase-3/7 activity detected using a fluorescent caspase-3/7 substrate. [1]

AIM-100 (4 mg/kg) reduces ataxia telangiectasia rupture (ATM) expression in amygdala-castrated nude mice, thereby inhibiting the formation of castrate-resistant radioresistant castrated obese carcinoma (CRPC) xenograft tumors [2].

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In castrated male nude mice, subcutaneous xenograft tumors were established by injecting LNCaP-caAck cells (2x10⁶ cells suspended in 100 μL PBS and 100 μL Matrigel). Mice were injected intraperitoneally (i.p.) with AIM-100 at a dose of 4 mg/kg body weight. The first injection was on day 7 post-cell injection, followed by five more injections on days 11, 15, 19, 23, and 27. AIM-100 treatment significantly inhibited the growth of unirradiated LNCaP-caAck xenograft tumors, resulting in approximately 50% reduction in tumor volume compared to vehicle-treated control. [2]
In a separate experiment, mice bearing irradiated (1.5 Gy) LNCaP-caAck xenografts were treated with AIM-100 (4 mg/kg, i.p.) on a similar schedule (starting day 7, with five total injections). The combination of irradiation and AIM-100 treatment resulted in significant suppression of tumor growth compared to irradiation alone or vehicle control. [2]
Immunohistochemical analysis of excised xenograft tumors from AIM-100-treated mice showed decreased ATM protein expression compared to tumors from vehicle-treated mice. [2]

Kinase assays were performed in the presence of increasing concentrations of AIM-100 to establish its specificity. The assays revealed that AIM-100 specifically inhibits Ack1 with an IC₅₀ of 22 nmol/L, but does not inhibit 30 other tested kinases, including PI3-kinase subfamily members and the three AKT isoforms. [1]

For cell proliferation assays (MTT assay), CD18, Panc-1, HPNE, OV90, MCF-7, MDA-MB-468, and MEF cells were untreated or treated with 2 to 10 μmol/L AIM-100 for 48 hours. The assay was performed twice with eight replicates. [1]
For Western blot analysis to assess protein phosphorylation, serum-depleted cells (CD18, Panc-1, MCF-7, H292, A2780-CP) were treated with insulin (0.8 μg/mL for 30 minutes) or EGF (10 ng/mL for 10 minutes) with or without pretreatment with AIM-100 (0.8-10 μmol/L overnight). Cell lysates were immunoblotted using antibodies against pTyr284-Ack1, pTyr176-AKT, pSer473-AKT, pThr308-AKT, panAKT, Ack1, and tubulin. Some lysates were immunoprecipitated with pTyr176-AKT antibody followed by immunoblotting with AKT antibody. [1]
For siRNA transfection, CD18 cells were electroporated with control or Ack1 siRNA. Cell lysates were immunoblotted with Ack1 and actin antibodies. Transfected cells were also assessed for cell proliferation by phase-contrast imaging and WST-1 assay. [1]
For cell cycle analysis, Panc-1 cells were untreated or treated with 6 μmol/L AIM-100 for 48 hours, stained with propidium iodide, and analyzed by flow cytometry. [1]
For apoptosis assay, Panc-1 cells were untreated or treated with 10 μmol/L AIM-100 for 48 hours. A caspase-3/7 green detection reagent was added, and caspase activity was assessed by monitoring green fluorescence under a microscope. The number of fluorescent-positive cells was counted in multiple fields. [1]

Male nude castrated mice were used for xenograft studies. LNCaP-caAck cells (2x10⁶ cells) were suspended in 100 μL of PBS and mixed with 100 μL of Matrigel. The cell suspension was injected subcutaneously into the flanks of the mice. [2]
For drug treatment, AIM-100 was dissolved in dimethyl sulfoxide (DMSO) and administered via intraperitoneal (i.p.) injection at a dose of 4 mg/kg body weight per injection. The treatment schedule began on day 7 after tumor cell injection. Mice received a total of six injections on days 7, 11, 15, 19, 23, and 27. Tumor volumes were measured twice weekly using calipers. [2]
In some experiments, tumor cells were irradiated (1.5 Gy) prior to injection into mice. The subsequent drug treatment schedule was the same. [2]
At the end of the experiment, mice were sacrificed, tumors were excised, fixed, paraffin-embedded, and sectioned for immunohistochemical analysis. [2]

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Male nude castrated mice were used for xenograft studies. LNCaP-caAck cells (2x10⁶ cells) were suspended in 100 μL of PBS and mixed with 100 μL of Matrigel. The cell suspension was injected subcutaneously into the flanks of the mice. [2]
For drug treatment, AIM-100 was dissolved in dimethyl sulfoxide (DMSO) and administered via intraperitoneal (i.p.) injection at a dose of 4 mg/kg body weight per injection. The treatment schedule began on day 7 after tumor cell injection. Mice received a total of six injections on days 7, 11, 15, 19, 23, and 27. Tumor volumes were measured twice weekly using calipers. [2]
In some experiments, tumor cells were irradiated (1.5 Gy) prior to injection into mice. The subsequent drug treatment schedule was the same. [2]
At the end of the experiment, mice were sacrificed, tumors were excised, fixed, paraffin-embedded, and sectioned for immunohistochemical analysis. [2]

[1]. Ack1 tyrosine kinase activation correlates with pancreatic cancer progression. Am J Pathol. 2012 Apr;180(4):1386-93.

[2]. Ack1-mediated androgen receptor phosphorylation modulates radiation resistance in castration-resistant prostate cancer. J Biol Chem. 2012 Jun 22;287(26):22112-22.

[3]. Effect of Ack1 tyrosine kinase inhibitor on ligand-independent androgen receptor activity. Prostate. 2010 Sep 1;70(12):1274-85.

AIM-100 is a 4-amino-5,6-biaryl-furo[2,3-d]pyrimidine derivative. [1]
AIM-100 was identified as a potent Ack1 inhibitor and has been shown to inhibit prostate cancer cell proliferation in previous studies. [1]
In this study, activated Ack1 (pTyr284-Ack1) expression was significantly upregulated in pancreatic intraepithelial neoplasia (PanIN) and metastatic pancreatic cancer, correlating with disease severity and poor patient survival. AIM-100 inhibits this pathway, suggesting its potential as a therapeutic agent for pancreatic cancer. [1]
The study highlights the importance of the Ack1/AKT signaling axis in pancreatic cancer progression and suggests that targeted inhibition of Ack1 by compounds like AIM-100 could be a promising therapeutic strategy. [1]

371.163

C, 74.37; H, 5.70; N, 11.31; O, 8.61

873305-35-2

11501591

White to yellow solid powder

5.22

1

5

5

28

492

1

C1C[C@H](OC1)CNC2=C3C(=C(OC3=NC=N2)C4=CC=CC=C4)C5=CC=CC=C5

XNFHHOXCDUAYSR-SFHVURJKSA-N

InChI=1S/C23H21N3O2/c1-3-8-16(9-4-1)19-20-22(24-14-18-12-7-13-27-18)25-15-26-23(20)28-21(19)17-10-5-2-6-11-17/h1-6,8-11,15,18H,7,12-14H2,(H,24,25,26)/t18-/m0/s1

N-[[(2S)-oxolan-2-yl]methyl]-5,6-diphenylfuro[2,3-d]pyrimidin-4-amine

AIM-100 AIM100 AIM 100

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)

DMSO : ~50 mg/mL (~134.61 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.73 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 (6.73 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 25.0 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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Solubility in Formulation 3: ≥ 2.5 mg/mL (6.73 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.)