PF-06751979
目录号 : GC30876
PF-06751979是一种有效的，脑渗透性的β-淀粉样前体蛋白裂解酶1(BACE1)抑制剂，IC50为7.3nM。
Cas No.:1818339-66-0
Sample solution is provided at 25 µL, 10mM.
;)
,-1-7$$$37316672.jpg');)
;)
;)
$$$36806353.jpg');)
;33(7)%EF%BC%9A546-561$$$37156877.jpg');)
;)
;)
;)
%201124-1138.$$$35908550.jpg');)
%20766-780.$$$37100057.jpg');)
%20418-432.$$$36893736.jpg');)
%EF%BC%9A-240-255.$$$35108512.jpg');)
533-545$$$36543525.jpg');)
%201-13.$$$36600053.jpg');)
;)
Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Kinase experiment: | Both BACE1 and BACE2 enzymatic activity is measured with the aid of an optimized synthetic peptide substrate biotin-GLTNIKTEEISEISYEVEFR-C[Oregon green]KK-OH. Upon cleavage of the peptide substrate, a decrease in fluorescence polarization is measured. Compounds are diluted by half log in 100% DMSO 11 times with a top concentration of 10mM in a 384-well polypropylene plate. The 100% DMSO dose response curve is then added to a 384-well black assay plate as 0.150 μL per well. The final working top concentration is 0.1 mM, and the DMSO concentration is 1%. A volume of 7.5 μL of BACE substrate is then added in assay buffer (100 mM sodium acetate, pH to 4.5 with glacial acetic acid, 0.001% Tween 20). The background wells in column 1 of the 384-well assay plate receive 7.5 μL of assay buffer. The reaction is started with the addition of 7.5 μL of BACE1 or BACE2 enzyme in assay buffer to all wells except the background wells in column 1. The final concentration of peptide substrate is 150 nM, and the final concentration of BACE1 and BACE2 enzyme is 0.15 and 2.5 nM, respectively. The assay plate is sealed and incubated at 37°C for 3 or 1 h (BACE1 or BACE2, respectively). After incubation, 15 μL of stop solution (1.5 μM streptavidin in Dulbecco’s PBS) is added to all wells, and the plate is read. Percent effect values for each concentration of compound are calculated based on fluorescence polarization (FP) readings in the 100% effect control wells containing no enzyme and the 0% effect control wells containing no compound. Curve-fitting analysis utilizing concentrations and percent effect values for a given compound is plotted, and the IC50 is determined using a sigmoidal four-parameter fit algorithm[1]. |
Cell experiment: | The neuroglioma cell line H4 cells are grown in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS) and 200 mM glutamax. Cells are plated overnight in tissue culture treated Falcon 384-well plates at a cell density of 4500 cells/well in 50 μL of media. The next day, media is removed, and cells are washed once with PBS, after which 25 μL of media is placed in all wells, followed by the addition of the diluted PF-06751979 dose response curve. The highest concentration tested is 30 μM with 1% DMSO. Cells serving as the background controls receive 30 μM of a proprietary compound. Compounds are allowed to incubate with cells overnight in a 37°C incubator. Concurrently, 384-well black Nunc Maxisorp plates are also incubated overnight at 4°C with 10 μL of 4 μg/mL Aβ antibody in coating buffer (0.1 M sodium bicarbonate, pH 8.8 to 9.0)[1]. |
Animal experiment: | Mice[1]Male 129/sve wild-type mice (20-25 g) are in a nonfasted state prior to subcutaneous dosing with vehicle or PF-06751979 using a dosing volume of 10 mL/kg. The mice are dosed subcutaneously once a day for 5 days with PF-06751979 (10 or 50 mg/kg/day) or vehicle. The mice (n=5 per group) are then sacrificed at 1, 3, 5, 7, 14, 20, and 30 h postdose. Following cardiac puncture into ethylenediaminetetraacetic acid (EDTA)-containing tubes, whole blood samples (0.5-1.0 mL) are collected, and plasma is separated by centrifugation (1500× g for 10 min at 4°C). The generated plasma is distributed into separate tubes on wet ice for exposure measurements (50 μL) and Aβ analysis (remainder). CSF samples (8-12 μL) are obtained by cisterna magna puncture using a sterile 25 gauge needle and collected with a P-20 Eppendorff pipet. CSF samples are distributed into separate tubes on dry ice for exposure measurements (3 μL) and Aβ analysis (remainder). Whole brain is removed and divided for exposure measurements (cerebellum) and Aβ analysis (left and right hemispheres), weighed, and frozen on dry ice. Prior to the assay, all samples are stored at -80°C[1]. |
References: [1]. O'Neill BT, et al. Design and Synthesis of Clinical Candidate PF-06751979: A Potent, Brain Penetrant, β-Site Amyloid Precursor Protein Cleaving Enzyme 1 (BACE1) Inhibitor Lacking Hypopigmentation. J Med Chem. 2018 May 24;61(10):4476-4504. | |
PF-06751979 is a potent, brain penetrant, β-site amyloid precursor protein cleaving enzyme 1 (BACE1) inhibitor with an IC50 of 7.3 nM in BACE1 binding assay.
PF-06751979 shows improved selectivity over BACE2 (IC50=194 nM) in binding (27-fold) relative to the literature examples and across multiple chemical series in BACE1 program. PF-06751979 also inhibits BACE1 and BACE2 in a fluorescent polarization (FP) assay with IC50s of 26.9 nM and 238 nM, respectively. PF-06751979 has excellent potency at BACE1 in binding or FP assay formats along with cellular activity looking at production of sAPPβ in H4 cells with an IC50 of 5 nM[1].
PF-06751979 displays excellent brain penetration, potent in vivo efficacy, and broad selectivity over related aspartyl proteases including BACE2. Acute administration of PF-06751979 yields a robust dose-responsive and time-dependent reduction of cerebral spinal fluid (CSF) Aβx-40 with peak inhibition at 3 h of >77%. To determine if the reduction in brain and CSF Aβ is maintained during sustained exposure to PF-06751979, a 5 day subchronic study is executed, dosing once daily by subcutaneous (SC) administration (10 or 50 mg/kg/day). Brain and CSF samples are collected on day 5, following the last dose. PF-06751979 produces a dose-responsive and time-dependent inhibition of Aβ42 in mouse brain. At the 50 mg/kg/day dose, maximal brain lowering is 63% at 7 to 9 h. Administration of PF-06751979 (10 or 50 mg/kg/day for 5 days) produces a dose-responsive and time-dependent inhibition of Aβx-40 in mouse CSF resulting in 77% inhibition of CSF at 3 h post-final 50 mg/kg dose[1].
[1]. O'Neill BT, et al. Design and Synthesis of Clinical Candidate PF-06751979: A Potent, Brain Penetrant, β-Site Amyloid Precursor Protein Cleaving Enzyme 1 (BACE1) Inhibitor Lacking Hypopigmentation. J Med Chem. 2018 May 24;61(10):4476-4504.
| Cas No. | 1818339-66-0 | SDF | |
| Canonical SMILES | FC(F)OC1=CN=C(C(NC2=CSC([C@]34CO[C@@H](C)C[C@@]3([H])CSC(N)=N4)=N2)=O)C=C1 | ||
| 分子式 | C18H19F2N5O3S2 | 分子量 | 455.5 |
| 溶解度 | DMSO : 150 mg/mL (329.31 mM);Ethanol : 50 mg/mL (109.77 mM) | 储存条件 | Store 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。 | ||
| 制备储备液 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 2.1954 mL | 10.9769 mL | 21.9539 mL |
| 5 mM | 0.4391 mL | 2.1954 mL | 4.3908 mL |
| 10 mM | 0.2195 mL | 1.0977 mL | 2.1954 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Safety, Tolerability, Pharmacokinetics, and Pharmacodynamic Effects of PF-06751979, a Potent and Selective Oral BACE1 Inhibitor: Results from Phase I Studies in Healthy Adults and Healthy Older Subjects
PF-06751979 is a selective inhibitor of the beta-site amyloid precursor protein cleaving enzyme-1, which is a key aspartyl protease in the generation of amyloid-β (Aβ) peptides, thought to be critical for the cerebral degeneration observed in Alzheimer's disease. Two Phase I studies (NCT02509117, NCT02793232) investigated the safety/tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) of PF-06751979. Single-ascending doses up to 540 mg and multiple-ascending doses up to 275 mg once daily (QD) in healthy adults, and multiple doses of 50 mg or 125 mg QD in healthy older subjects were assessed. PF-06751979 was well tolerated at all doses given, and all treatment-related adverse events (AEs) were mild to moderate. PK parameters remained consistent across the PF-06751979 QD dosing regimens, and no notable food effects were observed. PD analysis showed that PF-06751979 reduced the cerebrospinal fluid (CSF) and plasma levels of Aβ peptides in a dose-dependent manner, with the greatest reductions observed in subjects treated with 275 mg QD (approximately 92% and 93% reduction in CSF Aβ1-40 and Aβ1-42 observed at 24 h after Day 14 dose, respectively). A drug interaction study (NCT03126721) using midazolam indicated that there was no clinically meaningful effect of multiple doses of PF-06751979 100 mg QD on the PK of single-dose midazolam in healthy adults. Overall, these data suggest that PF-06751979 with daily dosing is favorable for further clinical development.
Design and Synthesis of Clinical Candidate PF-06751979: A Potent, Brain Penetrant, β-Site Amyloid Precursor Protein Cleaving Enzyme 1 (BACE1) Inhibitor Lacking Hypopigmentation
A major challenge in the development of β-site amyloid precursor protein cleaving enzyme 1 (BACE1) inhibitors for the treatment of Alzheimer's disease is the alignment of potency, drug-like properties, and selectivity over related aspartyl proteases such as Cathepsin D (CatD) and BACE2. The potential liabilities of inhibiting BACE2 chronically have only recently begun to emerge as BACE2 impacts the processing of the premelanosome protein (PMEL17) and disrupts melanosome morphology resulting in a depigmentation phenotype. Herein, we describe the identification of clinical candidate PF-06751979 (64), which displays excellent brain penetration, potent in vivo efficacy, and broad selectivity over related aspartyl proteases including BACE2. Chronic dosing of 64 for up to 9 months in dog did not reveal any observation of hair coat color (pigmentation) changes and suggests a key differentiator over current BACE1 inhibitors that are nonselective against BACE2 in later stage clinical development.
Selective Secretase Targeting for Alzheimer's Disease Therapy
Alzheimer's disease (AD) is associated with marked atrophy of the cerebral cortex and accumulation of amyloid plaques and neurofibrillary tangles. Amyloid plaques are formed by oligomers of amyloid-β (Aβ) in the brain, with a length of 42 and 40 amino acids. α-secretase cleaves amyloid-β protein precursor (AβPP) producing the membrane-bound fragment CTFα and the soluble fragment sAβPPα with neuroprotective activity; β-secretase produces membrane-bound fragment CTFβ and a soluble fragment sAβPPβ. After α-secretase cleavage of AβPP, γ-secretase cleaves CTFα to produce the cytoplasmic fragment AICD and P3 in the non-amyloidogenic pathway. CTFβ is cleaved by γ-secretase producing AICD as well as Aβ in amyloidogenic pathways. In the last years, the study of natural products and synthetic compounds, such as α-secretase activity enhancers, β-secretase inhibitors (BACE-1), and γ-secretase activity modulators, have been the focus of pharmaceuticals and researchers. Drugs were improved regarding solubility, blood-brain barrier penetration, selectivity, and potency decreasing Aβ42. In this regard, BACE-1 inhibitors, such as Atabecestat, NB-360, Umibecestat, PF-06751979 Verubecestat, LY2886721, Lanabecestat, LY2811376 and Elenbecestat, were submitted to phase I-III clinical trials. However, inhibition of Aβ production did not recover cognitive functions or reverse disease progress. Novel strategies are being developed, aiming at a partial reduction of Aβ production, such as the development of γ-secretase modulators or α-secretase activity enhancers. Such therapeutic tools shall focus on slowing down or minimizing the progression of neuronal damage. Here, we summarize structures and activities of the latest compounds designed for AD treatment, with remarkable in vitro, in vivo, and clinical phase activities.