PI3Kγ (IC50 = 9 nM); PI3Kα (IC50 = 39 nM); PI3Kδ (IC50 = 43 nM); PI3Kβ (IC50 = 113 nM); DNA-PK (IC50 = 150 nM)

PI3K-IN-1 (25 μM) blocks PI3K/Akt signaling pathways. With relatively lower expressions of -SMA, Col-1, and Timp-1, TGF-1-induced transformation into myofibroblast is also inhibited by the PI3K inhibitor PI3K-IN-1.

In mice models, BxPC-3 xenograft growth is significantly inhibited by the combination of XL765 (30 mg/kg) and chloroquine (50 mg/kg), whereas XL765 alone at the same dose has no inhibitory effect. [2]
In nude mice implanted intracranially with GBM 39-luc cells, oral administration of XL765 results in a greater than 12-fold reduction in median tumor bioluminescence compared to control and an improvement in median survival. In comparison to temozolomide (TMZ) alone, the combination of XL765 and TMZ results in a 140-fold reduction in median bioluminescence and a slight improvement in median survival. [3]

Kinase activity for PI3K isoforms is measured as the percentage of ATP consumed following the kinase reaction using luciferase-luciferin-coupled chemiluminescence, with ATP concentrations approximately equal to the Kmfor each respective kinase. Kinase reactions are initiated by combining test compounds, ATP and kinase in a 20 μL volume. PI3Kα, PI3Kβ, PI3Kγ, and PI3Kδ final enzyme concentrations are 0.5, 8, 20, and 2 nM, respectively. A similar assay format is used for DNA-PK (DNA protein kinase) and VPS34. VPS34 assay buffer contains 20 mM Tris-HCl, pH 7.5, 3.5 mM MnCl2, 100 mM NaCl, 1 mM DTT, and 0.01% cholamidopropyldimethylammonio propanesulfonate (CHAPS). Of note, 0.5 μL DMSO containing varying concentrations of the test compound is mixed with 10 μL enzyme solution (2× concentration). Kinase reactions are initiated by the addition of 10 μL of liver phosphatidylinositol and ATP solution (2× concentration). Assay concentrations for VPS34, ATP, and phosphatidylinositol are 40 nM, 1 μM, and 5 μM, respectively.

Cells (Pancreatic cancer cell lines (HcG25, Panc89, PA-TU8988T, Panc2.13, MiaPaCa2, Panc10.05, Panc8.13, BxPC-3, etc.) are treated with XL765 24 hours after plating and harvested for apoptosis or autophagy assays at 24, 48, or 72 hours after XL765 treatment. Apoptosis is determined by total percentage of annexin V-positive cells by fluorescence-activated cell sorting (FACS).Acidic vesicular organelles (AVOs) are detected in XL765-treated cells by vital staining with acridine orange. The degree of AVO formation is expressed as fold increase of acridine orange fluorescence intensity (FL3) in XL765-treated cells versus control cells.XL765 is active against class I PI3K (IC50 = 39, 113, 9 and 43 nM for p110α, β, γ and δ, respectively). XL765 also inhibits DNA-PK (IC50 = 150 nM) and mTOR (IC50 = 157 nM) but not XL-147 which shows IC50 values of > 15 μM.XL765 treatment results in decreased cell viability in 13 PDA cell lines in a dose-dependent manner. XL765, a dual-target PI3K/mTOR inhibitor, inhibits cell growth and apoptosis in many more cell lines and at lower concentrations as compared to the PI3K-selective inhibitors XL147 and PIK90. The effect can be recapitulated by using combinations of single-targeted compounds. XL765 significantly reduces phosphorylation of the mTOR targets S6, S6K, and 4EBP1, which is associated with greater apoptosis induction rather than to PI3K inhibition alone. XL765 treatment causes accumulation of autophagosomes in MIAPaCa-2 cells, and results in significant dose-dependent AVO induction and LC3-II stimulation in MIAPaCa-2 cells stably expressing a LC3-GFP construct.

Female Nu/Nu mice inoculated s.c. with BxPC-3 cells
30 mg/kg
Oral gavage once a day

Absorption, Distribution and Excretion
Orally available

[1]. Oncogene. 2008 Sep 18;27(41):5511-26.

[2]. J Mol Med (Berl). 2011 Sep;89(9):877-89.

[3]. Neuro Oncol. 2011 Apr;13(4):384-92.

XL765 is a sulfonamide obtained by formal condensation of the sulfonic acid group of 4-[(3-methoxy-4-methylbenzoyl)amino]benzenesulfonic acid with the primary aromatic amino group of N-(3,5-dimethoxyphenyl)quinoxaline-2,3-diamine. A dual PI3K/mTOR inhibitor used in cancer treatment. It has a role as an EC 2.7.1.137 (phosphatidylinositol 3-kinase) inhibitor, an antineoplastic agent and a mTOR inhibitor. It is a sulfonamide, a quinoxaline derivative, an aromatic amine, a member of benzamides and an aromatic ether.
XL765 is an orally available small molecule that has been shown in preclinical studies to selectively inhibit the activity of phosphoinositide-3 kinase (PI3K) and mammalian target of rapamycin (mTOR). It is being developed by Exelixis, Inc.

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Drug Indication
For the treatment of various forms of cancer.

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Mechanism of Action
XL765 inhibits the activity of phosphoinositide-3 kinase (PI3K), which is frequently activated in tumors and promotes cell growth, survival and resistance to chemotherapy and radiotherapy. XL765 also inhibits mammalian target of rapamycin (mTOR), which also is activated frequently in human tumors and plays a central role in tumor cell growth. XL765 potently inhibits Class I PI3K isoforms and mTOR.

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Pharmacodynamics
XL765 is an orally available small molecule that has been shown in preclinical studies to selectively inhibit the activity of phosphoinositide-3 kinase (PI3K) and mammalian target of rapamycin (mTOR). Activation of PI3K is a frequent event in human tumors, promoting tumor cell growth, survival, and resistance to chemotherapy and radiotherapy. mTOR is also frequently activated in human tumors and plays a central role in tumor cell growth. mTOR can be activated via upregulation of PI3K, or via PI3K-independent mechanisms. Inactivation of PI3K has been shown to inhibit growth and induce apoptosis (programmed cell death) in tumor cells, whereas inactivation of mTOR has been shown to inhibit the growth of tumor cells. In preclinical studies, XL765 slowed tumor growth or caused tumor shrinkage in multiple preclinical cancer models, including breast, lung, ovarian, and prostate cancers, and gliomas. XL765 has also been shown to enhance the anti-tumor effects of several chemotherapeutic agents in preclinical cancer models. XL765 has at least a 24-hour duration of action against both PI3K and mTOR readouts in tumors in vivo following a single oral (PO) dose of 30-100 mg/kg.

C31H29N5O6S

599.66

599.183

C, 62.09; H, 4.87; N, 11.68; O, 16.01; S, 5.35

1349796-36-6

1349796-36-6;934493-76-2;

49867926

Light yellow to yellow solid powder

1.4±0.1 g/cm3

1.674

5.96

3

10

10

43

993

0

S(C1C([H])=C([H])C(=C([H])C=1[H])N([H])C(C1C([H])=C([H])C(C([H])([H])[H])=C(C=1[H])OC([H])([H])[H])=O)(N([H])C1C(=NC2=C([H])C([H])=C([H])C([H])=C2N=1)N([H])C1C([H])=C(C([H])=C(C=1[H])OC([H])([H])[H])OC([H])([H])[H])(=O)=O

HJSSPYJVWLTYHG-UHFFFAOYSA-N

InChI=1S/C31H29N5O6S/c1-19-9-10-20(15-28(19)42-4)31(37)33-21-11-13-25(14-12-21)43(38,39)36-30-29(34-26-7-5-6-8-27(26)35-30)32-22-16-23(40-2)18-24(17-22)41-3/h5-18H,1-4H3,(H,32,34)(H,33,37)(H,35,36)

N-[4-[[[3-[(3,5-dimethoxyphenyl)amino]-2-quinoxalinyl]amino]sulfonyl]phenyl]-3-methoxy-4-methyl-benzamide

SAR245409 analogue; SAR245409 analogue; SAR 245409analogue; XL765 analogue; XL 765 analogue; XL-765 analogue; Voxtalisib analogue

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: ~12 mg/mL (20.0 mM)
Water: <1 mg/mL
Ethanol: <1 mg/mL

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.17 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 (4.17 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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: 30% PEG400+0.5% Tween80+5%Propylene glycol: 30mg/mL

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