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Quality Control & SDS

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Purity: >99.50%

实验参考方法

Kinase experiment [1]:
Preparation Method	Inhibitor(GDC-0068 (RG7440)), enzyme (9 nM Akt1 or 100 pM PKA), and substrate (100 nM Crosstide) were incubated with 5 μM ATP in assay buffe, final DMSO 2% (v/v)) for 60 min at ambient temperature in a 5 μL reaction volume. Reactions were initiated by addition of enzyme + peptide substrate to ATP solutions. IMAP binding reagent (15 μL) was added to terminate the reaction, and the stopped reactions were incubated for a minimum of 30 min at room temperature (rt).
Applications	GDC-0068 (RG7440) is a selective, ATP-competitive pan-Akt inhibitor that inhibits Akt1, Akt2, and Akt3 at an IC50 of 5,18,8 nM, respectively.
Cell experiment [2]:
Cell lines	HCT116 cell
Preparation Method	To study how GDC-0068 (RG7440) influences tumor progression, cell viability was detected by CCK-8 in HCT116 at indicated time points after 1–20 μmol/L GDC-0068 (RG7440)treatment.
Reaction Conditions	1–20 μmol/L GDC-0068 (RG7440) for 0, 3, 6, 12, and 24 h
Applications	Cell viability decreased markedly with increasing dose or time, cell proliferation was inhibited by GDC-0068 (RG7440) in a dose- and time-dependent manner.
Animal experiment [3]:
Animal models	Nude mice
Preparation Method	Nude mice were injected s.c. with HCT116 WT or PUMA−/−. Once the tumor was measurable, mice were treated daily with 30 mg/kg GDC-0068 (RG7440) by oral gavage, for 15 consecutive days.
Dosage form	30 mg/kg GDC-0068 (RG7440) by oral gavage, for 15 consecutive days
Applications	GDC-0068 (RG7440) therapy significantly inhibited the growth of WT tumors, however, for the PUMA / tumors, GDC-0068 (RG7440) therapy caused some suppression, but not so markedly compared with that of WT tumors
References: [1]. Blake JF, Xu R, et,al. Discovery and preclinical pharmacology of a selective ATP-competitive Akt inhibitor (GDC-0068) for the treatment of human tumors. J Med Chem. 2012 Sep 27;55(18):8110-27. doi: 10.1021/jm301024w. Epub 2012 Sep 18. PMID: 22934575. [2]. Sun L, Huang Y, et,al.Ipatasertib, a novel Akt inhibitor, induces transcription factor FoxO3a and NF-κB directly regulates PUMA-dependent apoptosis. Cell Death Dis. 2018 Sep 5;9(9):911. doi: 10.1038/s41419-018-0943-9. PMID: 30185800; PMCID: PMC6125489.

产品描述

GDC-0068 (RG7440, Ipatasertib) is a novel highly selective ATP-competitive pan-Akt inhibitor that inhibits all three isoforms of Akt1, Akt2, and Akt3 at an IC50 of 5,18,8 nM, respectively ^[1,5].

GDC-0068 decreased cell viability, induced apoptosis, and inhibited phosphorylation of proline rich Akt substrate 40 kDa and p70 S6 kinase in a dose-dependent manner in PIK3CA-mutant breast cancer brain metastatic cell lines compared with PIK3CA-wildtype cell lines^[6]. The combined treatment of GDC-0068 (RG7440) plus anti-microtubule chemotherapy, including vinorelbine, paclitaxel, eribulin, is contributed to anti-proliferative, pro-apoptotic, and anti-metastatic effect on human breast cancer cells^[7]. GDC-0068 (RG7440) blocked Akt signaling both in cultured human cancer cell lines and in tumor xenograft models as evidenced by dose-dependent decrease in phosphorylation of downstream targets. Inhibition of Akt activity by GDC-0068 (RG7440) resulted in blockade of cell-cycle progression and reduced viability of cancer cell lines^[2].

Preclinical Pharmacology demonstrates GDC-0068 (RG7440) exposure resulting in a dose-dependently pharmacodynamic effect and robust anti-tumor activity in a broad spectrum of human cancer cells in vitro and in vivo^[5]. GDC-0068 (RG7440) therapy significantly inhibited the growth of WT tumors, however, for the PUMA / tumors, GDC-0068 (RG7440) therapy caused some suppression, but not so markedly compared with that of WT tumors. Immunohistochemistry staining shows that the expression of P-Akt reduced in both WT and PUMA / tumors after GDC-0068 (RG7440) therapy. PUMA-dependent antitumor effects of GDC-0068 (RG7440) in colon cancer^[4]. As an ATP-competitive Akt inhibitor, GDC-0068 (RG7440) is a strong and safe target for Akt, which has been proved in a first-in-human phase I study^[3].

References:
[1]. Lin K, Lin J, et,al. An ATP-site on-off switch that restricts phosphatase accessibility of Akt. Sci Signal. 2012 May 8;5(223):ra37. doi: 10.1126/scisignal.2002618. PMID: 22569334.
[2]. Lin J, Sampath D, et,al.Targeting activated Akt with GDC-0068, a novel selective Akt inhibitor that is efficacious in multiple tumor models. Clin Cancer Res. 2013 Apr 1;19(7):1760-72. doi: 10.1158/1078-0432.CCR-12-3072. Epub 2013 Jan 3. PMID: 23287563.
[3]. Saura C, Roda D, et,al. A First-in-Human Phase I Study of the ATP-Competitive AKT Inhibitor Ipatasertib Demonstrates Robust and Safe Targeting of AKT in Patients with Solid Tumors. Cancer Discov. 2017 Jan;7(1):102-113. doi: 10.1158/2159-8290.CD-16-0512. Epub 2016 Nov 21. Erratum in: Cancer Discov. 2018 Nov;8(11):1490. PMID: 27872130; PMCID: PMC5463454.
[4]. Sun L, Huang Y, et,al. Ipatasertib, a novel Akt inhibitor, induces transcription factor FoxO3a and NF-κB directly regulates PUMA-dependent apoptosis. Cell Death Dis. 2018 Sep 5;9(9):911. doi: 10.1038/s41419-018-0943-9. PMID: 30185800; PMCID: PMC6125489.
[5]. Blake JF, Xu R, et,al. Discovery and preclinical pharmacology of a selective ATP-competitive Akt inhibitor (GDC-0068) for the treatment of human tumors. J Med Chem. 2012 Sep 27;55(18):8110-27. doi: 10.1021/jm301024w. Epub 2012 Sep 18. PMID: 22934575.
[6]. Ippen FM, Grosch JK, et,al. Targeting the PI3K/Akt/mTOR pathway with the pan-Akt inhibitor GDC-0068 in PIK3CA-mutant breast cancer brain metastases. Neuro Oncol. 2019 Nov 4;21(11):1401-1411. doi: 10.1093/neuonc/noz105. PMID: 31173106; PMCID: PMC6827829.
[7]. Yan Y, Serra V, et,al.Evaluation and clinical analyses of downstream targets of the Akt inhibitor GDC-0068. Clin Cancer Res. 2013 Dec 15;19(24):6976-86. doi: 10.1158/1078-0432.CCR-13-0978. Epub 2013 Oct 18. PMID: 24141624.

GDC-0068 (RG7440, Ipatasertib) 是一种新型高选择性 ATP 竞争性泛 Akt 抑制剂，可抑制 Akt1、Akt2 和 Akt3 的所有三种亚型，IC50 分别为 5、18、8 nM ^[1,5].

与 PIK3CA 野生型细胞系相比，GDC-0068 在 PIK3CA 突变型乳腺癌脑转移细胞系中以剂量依赖性方式降低细胞活力、诱导细胞凋亡并抑制富含脯氨酸的 Akt 底物 40 kDa 和 p70 S6 激酶的磷酸化^[6]。 GDC-0068 (RG7440) 联合抗微管化疗，包括长春瑞滨、紫杉醇、艾日布林，有助于对人乳腺癌细胞产生抗增殖、促凋亡和抗转移作用^[7]。 GDC-0068 (RG7440) 在培养的人类癌细胞系和肿瘤异种移植模型中阻断 Akt 信号，下游靶标磷酸化的剂量依赖性降低证明了这一点。 GDC-0068 (RG7440) 对 Akt 活性的抑制导致细胞周期进程受阻并降低癌细胞系的活力^[2]。

临床前药理学证明 GDC-0068 (RG7440) 暴露会在体外和体内的广谱人类癌细胞中产生剂量依赖性药效学效应和强大的抗肿瘤活性^[5] . GDC-0068（RG7440）治疗显着抑制WT肿瘤的生长，然而，对于PUMA/肿瘤，GDC-0068（RG7440）治疗引起一些抑制，但与WT肿瘤相比没有那么显着。免疫组织化学染色显示，在 GDC-0068 (RG7440) 治疗后，P-Akt 的表达在 WT 和 PUMA/肿瘤中均降低。 GDC-0068 (RG7440) 在结肠癌中的 PUMA 依赖性抗肿瘤作用^[4]。作为一种 ATP 竞争性 Akt 抑制剂，GDC-0068 (RG7440) 是 Akt 的一个强大而安全的靶点，这一点已在人体第一阶段研究中得到证实^[3]。

Chemical Properties

Cas No.	1001264-89-6	SDF
别名	帕他色替,GDC0068,RG7440,CS0975
化学名	(2S)-2-(4-chlorophenyl)-1-[4-[(5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl]piperazin-1-yl]-3-(propan-2-ylamino)propan-1-one
Canonical SMILES	CC1CC(C2=C1C(=NC=N2)N3CCN(CC3)C(=O)C(CNC(C)C)C4=CC=C(C=C4)Cl)O
分子式	C₂₄H₃₂ClN₅O₂	分子量	458
溶解度	≥ 22.9mg/mL in DMSO	储存条件	Desiccate at -20°C
General tips	请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。
Shipping Condition	评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。

溶解性数据

制备储备液
		1 mg	5 mg	10 mg
	1 mM	2.1834 mL	10.917 mL	21.8341 mL
	5 mM	0.4367 mL	2.1834 mL	4.3668 mL
	10 mM	0.2183 mL	1.0917 mL	2.1834 mL

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Research Update

Targeting the PI3K/Akt/mTOR pathway with the pan-Akt inhibitor GDC-0068 in PIK3CA-mutant breast cancer brain metastases

Neuro Oncol.2019 Nov 4;21(11):1401-1411. PMID: 31173106 DOI: 10.1093/neuonc/noz105

Background: Activating mutations in the pathway of phosphatidylinositol-3 kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) occur in 43-70% of breast cancer brain metastasis patients. To date, the treatment of these patients presents an ongoing challenge, mainly because of the lack of targeted agents that are able to sufficiently penetrate the blood-brain barrier. GDC-0068 is a pan-Akt inhibitor that has shown to be effective in various preclinical tumor models as well as in clinical trials. The purpose of this study was to analyze the efficacy of GDC-0068 in a breast cancer brain metastases model. Methods: In in vitro studies, antitumor activity of GDC-0068 was assessed in breast cancer cells of phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA)-mutant and PIK3CA-wildtype breast cancer cell lines using cell viability and apoptosis assays, cell cycle analysis, and western blots. In vivo, the efficacy of GDC-0068 was analyzed in a PIK3CA-mutant breast cancer brain metastasis orthotopic xenograft mouse model and evaluated by repeated bioluminescent imaging and immunohistochemistry. Results: GDC-0068 decreased cell viability, induced apoptosis, and inhibited phosphorylation of proline rich Akt substrate 40 kDa and p70 S6 kinase in a dose-dependent manner in PIK3CA-mutant breast cancer brain metastatic cell lines compared with PIK3CA-wildtype cell lines. In vivo, treatment with GDC-0068 notably inhibited the growth of PIK3CA-mutant tumors and resulted in a significant survival benefit compared with sham, whereas no effect was detected in a PIK3CA-wildtype model. Conclusions: This study suggests that the Akt inhibitor GDC-0068 may be an encouraging targeted treatment strategy for breast cancer brain metastasis patients with activating mutations in the PI3K pathway. These data provide a rationale to further evaluate the efficacy of GDC-0068 in patients with brain metastases.

A phase Ib open-label dose escalation study of the safety, pharmacokinetics, and pharmacodynamics of cobimetinib (GDC-0973) and ipatasertib (GDC-0068) in patients with locally advanced or metastatic solid tumors

Invest New Drugs.2021 Feb;39(1):163-174.PMID: 32737717DOI: 10.1007/s10637-020-00975-6

Background: This Phase Ib study explored combination dosing of the allosteric MEK1/2 inhibitor cobimetinib and the ATP-competitive pan-AKT inhibitor ipatasertib. Methods: Patients with advanced solid tumors were enrolled to two dose escalation arms, each using a 3 + 3 design in 28-day cycles. In Arm A, patients received concurrent cobimetinib and ipatasertib on days 1-21. In Arm B, cobimetinib was administered intermittently with ipatasertib for 21 days. Primary objectives evaluated dose-limiting toxicities (DLTs), maximum tolerated doses (MTD), and the recommended Phase II dose (RP2D). Secondary objectives included analysis of pharmacokinetic parameters, MAPK and PI3K pathway alterations, changes in tissue biomarkers, and preliminary anti-tumor efficacy. Expansion cohorts included patients with PTEN-deficient triple-negative breast cancer and endometrial cancer. Results: Among 66 patients who received ≥1 dose of study drug, all experienced an adverse event (AE). Although no DLTs were reported, 6 patients experienced Cycle 1 DLT-equivalent AEs. The most common treatment-related AEs were diarrhea, nausea, vomiting, dermatitis acneiform, and fatigue. Thirty-five (53%) patients experienced drug-related AEs of ≥ grade 3 severity. Cobimetinb/ipatasertib MTDs were 60/200 mg on Arm A and 150/300 mg on Arm B; the latter was chosen as the RP2D. No pharmacokinetic interactions were identified. Biomarker analyses indicated pathway blockade and increases in IFNγ and PD-L1 gene expression following the combination. Three patients with endometrial or ovarian cancer achieved partial response, all with PTEN-low disease and two with tumor also harboring KRAS mutation. Conclusion: There was limited tolerability and efficacy for this MEK and AKT inhibitor combination. Nonetheless, pharmacodynamic analyses indicated target engagement and suggest rationale for further exploration of cobimetinib or ipatasertib in combination with other anticancer agents. ClinicalTrials.gov identifier: NCT01562275.

Ipatasertib (GDC-0068) and erdafitinib co-treatment for inducing mitochondrial apoptosis through Bim upregulation in bladder cancer cells

Biochem Biophys Res Commun.2022 May 14;604:165-171.PMID: 35306249DOI: 10.1016/j.bbrc.2022.03.029

Bladder cancer (BC) is a common malignancy of the urological system that still lacks effective treatment. It is frequently characterised by dysregulation of fibroblast growth factor receptor (FGFR) signalling. FGFR inhibitors have been proven as a promising treatment for BC in clinical settings. Besides the FGFR signalling, the therapeutic effects of FGFR inhibitors are often limited owing to various mechanisms, such as the activation of the Akt signalling pathway. Therefore, this study aimed to examine the synergistic effects of ipatasertib, a FGFR inhibitor, and erdafitinib, an Akt inhibitor, in BC cells. Ipatasertib and erdafitinib co-treatment synergistically inhibited cell proliferation and induced BC cell death. Mechanically, ipatasertib and erdafitinib induced the activation of Bax, an essential protein for cell death. Moreover, erdafitinib, which inhibited the Akt signalling pathway, is responsible for Bim upregulation, a condition critical to achieving the synergistic effects. Therefore, our data suggest that ipatasertib and erdafitinib co-treatment is a promising strategy for BC.

Targeting activated Akt with GDC-0068, a novel selective Akt inhibitor that is efficacious in multiple tumor models

Clin Cancer Res.2013 Apr 1;19(7):1760-72.PMID: 23287563DOI: 10.1158/1078-0432.CCR-12-3072

Purpose: We describe the preclinical pharmacology and antitumor activity of GDC-0068, a novel highly selective ATP-competitive pan-Akt inhibitor currently in clinical trials for the treatment of human cancers. Experimental design: The effect of GDC-0068 on Akt signaling was characterized using specific biomarkers of the Akt pathway, and response to GDC-0068 was evaluated in human cancer cell lines and xenograft models with various genetic backgrounds, either as a single agent or in combination with chemotherapeutic agents. Results: GDC-0068 blocked Akt signaling both in cultured human cancer cell lines and in tumor xenograft models as evidenced by dose-dependent decrease in phosphorylation of downstream targets. Inhibition of Akt activity by GDC-0068 resulted in blockade of cell-cycle progression and reduced viability of cancer cell lines. Markers of Akt activation, including high-basal phospho-Akt levels, PTEN loss, and PIK3CA kinase domain mutations, correlate with sensitivity to GDC-0068. Isogenic PTEN knockout also sensitized MCF10A cells to GDC-0068. In multiple tumor xenograft models, oral administration of GDC-0068 resulted in antitumor activity ranging from tumor growth delay to regression. Consistent with the role of Akt in a survival pathway, GDC-0068 also enhanced antitumor activity of classic chemotherapeutic agents. Conclusions: GDC-0068 is a highly selective, orally bioavailable Akt kinase inhibitor that shows pharmacodynamic inhibition of Akt signaling and robust antitumor activity in human cancer cells in vitro and in vivo. Our preclinical data provide a strong mechanistic rationale to evaluate GDC-0068 in cancers with activated Akt signaling.

Evaluation and clinical analyses of downstream targets of the Akt inhibitor GDC-0068

Clin Cancer Res.2013 Dec 15;19(24):6976-86.DOI: 10.1158/1078-0432.CCR-13-0978

Purpose: The oncogenic PI3K/Akt/mTOR pathway is an attractive therapeutic target in cancer. However, it is unknown whether the pathway blockade required for tumor growth inhibition is clinically achievable. Therefore, we conducted pharmacodynamic studies with GDC-0068, an ATP competitive, selective Akt1/2/3 inhibitor, in preclinical models and in patients treated with this compound. Experimental design: We used a reverse phase protein array (RPPA) platform to identify a biomarker set indicative of Akt inhibition in cell lines and human-tumor xenografts, and correlated the degree of pathway inhibition with antitumor activity. Akt pathway activity was measured using this biomarker set in pre- and post-dose tumor biopsies from patients treated with GDC-0068 in the dose escalation clinical trial. Results: The set of biomarkers of Akt inhibition is composed of 10 phosphoproteins, including Akt and PRAS40, and is modulated in a dose-dependent fashion, both in vitro and in vivo. In human-tumor xenografts, this dose dependency significantly correlated with tumor growth inhibition. Tumor biopsies from patients treated with GDC-0068 at clinically achievable doses attained a degree of biomarker inhibition that correlated with tumor growth inhibition in preclinical models. In these clinical samples, compensatory feedback activation of ERK and HER3 was observed, consistent with preclinical observations. Conclusion: This study identified a set of biomarkers of Akt inhibition that can be used in the clinical setting to assess target engagement. Here, it was used to show that robust Akt inhibition in tumors from patients treated with GDC-0068 is achievable, supporting the clinical development of this compound in defined patient populations.

GDC-0068 (RG7440)

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