Evobrutinib (M2951)
(Synonyms: 埃沃布鲁替尼) 目录号 : GC34062
Evobrutinib 作为一种口服、高选择性、共价的布鲁顿氏酪氨酸激酶抑制剂，具有良好的耐受性和有效性。
Cas No.:1415823-73-2
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
B cells |
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Preparation Method |
In a complementary in vitro setting, freshly isolated B cells from non-evobrutinib-treated mice were directly pre-incubated with evobrutinib doses up to 1 µM. |
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Reaction Conditions |
1 µM; 30min |
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Applications |
Evobrutinib specifically inhibits B cellular excitatory calcium mobilization and cytokine production. |
| Animal experiment [2]: | |
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Animal models |
DBA/1J female mice aged 11–12 wk |
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Preparation Method |
Mice were administered a single dose of 12 mg/kg evobrutinib, and B cell inhibition was measured as before at different time points. |
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Dosage form |
12 mg/kg; p.o. |
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Applications |
16 h after dosing, B cell activation was still inhibited by roughly 50%. |
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References: [1]. Torke S, et al. Inhibition of Bruton's tyrosine kinase interferes with pathogenic B-cell development in inflammatory CNS demyelinating disease. Acta Neuropathol. 2020 Oct;140(4):535-548. [2]. Haselmayer P, et al. Efficacy and Pharmacodynamic Modeling of the BTK Inhibitor Evobrutinib in Autoimmune Disease Models. J Immunol. 2019 May 15;202(10):2888-2906. |
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Evobrutinib, as an orally, highly selective, covalent Bruton's tyrosine kinase inhibitor, was well‐tolerated and effective.[1] Evobrutinib was metabolized via hydroxylation, hydrolysis, O-dealkylation, glucuronidation, and GSH conjugation.[4]
n vitro efficacy test it shown that evobrutinib inhibits Btk in vitro with IC50 values in two studies 58 nM and 38 nM. Evobrutinib at concentrations of 10 μM did not increase bleeding time in vitro. [5] In U937 NF-κB–Luc reporter cells, evobrutinib inhibited NF-κB activation and, FcγR signaling with an IC50 of 78 nM. Evobrutinib inhibited BTK and BMX with IC50 values of 0.058 μM and 0.02 μM, respectively. Evobrutinib inhibited B cell activation in PBMCs with a mean IC50 of 15.8 nM. Evobrutinib inhibited basophil activation with an average IC50 of 1.66 μM. [2] In vitro, treatment with 100 to 1000 nM evobrutinib dose-dependently reduced calcium mobilization upon BCR ligation in a manner indistinguishable from the murine setting, while T cells again remained unaffected. In addition, production of IL-6, IFN-γ and IL-10 upon BCR ligation was reduced by 1000 nM evobrutinib.[3]
In vivo, treatment with 1, 3 or 10 mg/kg evobrutinib in C57/BL6 mice orally inhibited expression of molecules involved in B-cell antigen presentation. Evobrutinib treatment (10 mg/kg) functionally impaired the capacity of B cells to act as antigen-presenting cells for the development of encephalitogenic T cells, resulting in a remarkably decreased disease severity in mice.[3]
References:
[1]Scheible H, et al. Evobrutinib, a covalent Bruton's tyrosine kinase inhibitor: Mass balance, elimination route, and metabolism in healthy participants. Clin Transl Sci. 2021 Nov;14(6):2420-2430.?
[2]Haselmayer P, et al. Efficacy and Pharmacodynamic Modeling of the BTK Inhibitor Evobrutinib in Autoimmune Disease Models. J Immunol. 2019 May 15;202(10):2888-2906.
[3]Torke S, et al. Inhibition of Bruton's tyrosine kinase interferes with pathogenic B-cell development in inflammatory CNS demyelinating disease. Acta Neuropathol. 2020 Oct;140(4):535-548.?
[4]Li Z, et al. Identification of metabolites of evobrutinib in rat and human hepatocytes by using ultra-high performance liquid chromatography coupled with diode array detector and Q Exactive Orbitrap tandem mass spectrometry. Drug Test Anal. 2019 Jan;11(1):129-139.?
[5]von Hundelshausen P, et al. Bleeding by Bruton Tyrosine Kinase-Inhibitors: Dependency on Drug Type and Disease. Cancers (Basel). 2021 Mar 4;13(5):1103.
Evobrutinib 作为一种口服、高选择性、共价的布鲁顿氏酪氨酸激酶抑制剂，具有良好的耐受性和有效性。[1]Evobrutinib 通过羟基化、水解、O-脱烷基化、葡萄糖醛酸化和GSH 结合。[4]
体外药效试验表明，evobrutinib 在体外抑制 Btk，两项研究中的 IC50 值为 58 nM 和 38 nM。浓度为 10 μM 的 Evobrutinib 在体外不会增加出血时间。 [5] 在 U937 NF-κB-Luc 报告细胞中，evobrutinib 抑制 NF-κB 活化和 FcγR 信号，IC50 为 78 nM。 Evobrutinib 抑制 BTK 和 BMX，IC50 值分别为 0.058 μM 和 0.02 μM。 Evobrutinib 抑制 PBMC 中的 B 细胞活化，平均 IC50 为 15.8 nM。 Evobrutinib 抑制嗜碱性粒细胞活化，平均 IC50 为 1.66 μM。 [2] 在体外，100 至 1000 nM evobrutinib 剂量依赖性地减少了 BCR 结扎时的钙动员，其方式与鼠类设置没有区别，而 T 细胞再次保持不受影响。此外，1000 nM evobrutinib 减少了 BCR 连接时 IL-6、IFN-γ 和 IL-10 的产生。[3]
在体内，C57/BL6 小鼠口服 1、3 或 10 mg/kg evobrutinib 可抑制参与 B 细胞抗原呈递的分子的表达。 Evobrutinib 治疗 (10 mg/kg) 在功能上削弱了 B 细胞作为抗原呈递细胞的能力，从而促进了脑炎性 T 细胞的发育，从而显着降低了小鼠的疾病严重程度。[3]
| Cas No. | 1415823-73-2 | SDF | |
| 别名 | 埃沃布鲁替尼 | ||
| Canonical SMILES | C=CC(N1CCC(CNC2=NC=NC(N)=C2C3=CC=C(OC4=CC=CC=C4)C=C3)CC1)=O | ||
| 分子式 | C25H27N5O2 | 分子量 | 429.51 |
| 溶解度 | DMSO : 6.4 mg/mL (14.90 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。 |
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| 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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.3282 mL | 11.6412 mL | 23.2823 mL |
| 5 mM | 0.4656 mL | 2.3282 mL | 4.6565 mL |
| 10 mM | 0.2328 mL | 1.1641 mL | 2.3282 mL |
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动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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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，
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1. 首先保证母液是澄清的；
2.
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Efficacy and Pharmacodynamic Modeling of the BTK Inhibitor Evobrutinib in Autoimmune Disease Models
J Immunol2019 May 15;202(10):2888-2906.PMID: 30988116DOI: 10.4049/jimmunol.1800583
Because of its role in mediating both B cell and Fc receptor signaling, Bruton's tyrosine kinase (BTK) is a promising target for the treatment of autoimmune diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). Evobrutinib is a novel, highly selective, irreversible BTK inhibitor that potently inhibits BCR- and Fc receptor-mediated signaling and, thus, subsequent activation and function of human B cells and innate immune cells such as monocytes and basophils. We evaluated evobrutinib in preclinical models of RA and SLE and characterized the relationship between BTK occupancy and inhibition of disease activity. In mouse models of RA and SLE, orally administered evobrutinib displayed robust efficacy, as demonstrated by reduction of disease severity and histological damage. In the SLE model, evobrutinib inhibited B cell activation, reduced autoantibody production and plasma cell numbers, and normalized B and T cell subsets. In the RA model, efficacy was achieved despite failure to reduce autoantibodies. Pharmacokinetic/pharmacodynamic modeling showed that mean BTK occupancy in blood cells of 80% was linked to near-complete disease inhibition in both RA and SLE mouse models. In addition, evobrutinib inhibited mast cell activation in a passive cutaneous anaphylaxis model. Thus, evobrutinib achieves efficacy by acting both on B cells and innate immune cells. Taken together, our data show that evobrutinib is a promising molecule for the chronic treatment of B cell-driven autoimmune disorders.
Placebo-Controlled Trial of an Oral BTK Inhibitor in Multiple Sclerosis
N Engl J Med2019 Jun 20;380(25):2406-2417.PMID: 31075187DOI: 10.1056/NEJMoa1901981
Background: Bruton's tyrosine kinase (BTK) regulates the functions of B cells and myeloid cells that are implicated in the pathogenesis of multiple sclerosis. Evobrutinib is a selective oral BTK inhibitor that has been shown to inhibit B-cell activation both in vitro and in vivo.
Methods: In this double-blind, randomized, phase 2 trial, we assigned patients with relapsing multiple sclerosis to one of five groups: placebo, evobrutinib (at a dose of 25 mg once daily, 75 mg once daily, or 75 mg twice daily), or open-label dimethyl fumarate (DMF) as a reference. The primary end point was the total (cumulative) number of gadolinium-enhancing lesions identified on T1-weighted magnetic resonance imaging at weeks 12, 16, 20, and 24. Key secondary end points included the annualized relapse rate and change from baseline in the score on the Expanded Disability Status Scale (EDSS).
Results: A total of 267 patients were randomly assigned to a trial group. The mean (±SD) total number of gadolinium-enhancing lesions during weeks 12 through 24 was 3.85±5.44 in the placebo group, 4.06±8.02 in the evobrutinib 25-mg group, 1.69±4.69 in the evobrutinib 75-mg once-daily group, 1.15±3.70 in the evobrutinib 75-mg twice-daily group, and 4.78±22.05 in the DMF group. The baseline adjusted rate ratios for the total number of lesions over time as compared with placebo were 1.45 in the evobrutinib 25-mg group (P = 0.32), 0.30 in the evobrutinib 75-mg once-daily group (P = 0.005), and 0.44 in the evobrutinib 75-mg twice-daily group (P = 0.06). The unadjusted annualized relapse rate at week 24 was 0.37 in the placebo group, 0.57 in the evobrutinib 25-mg group, 0.13 in the evobrutinib 75-mg once-daily group, 0.08 in the evobrutinib 75-mg twice-daily group, and 0.20 in the DMF group. There was no significant effect of trial group on the change from baseline in the EDSS score. Elevations in liver aminotransferase values were observed with evobrutinib.
Conclusions: Patients with relapsing multiple sclerosis who received 75 mg of evobrutinib once daily had significantly fewer enhancing lesions during weeks 12 through 24 than those who received placebo. There was no significant difference with placebo for either the 25-mg once-daily or 75-mg twice-daily dose of evobrutinib, nor in the annualized relapse rate or disability progression at any dose. Longer and larger trials are required to determine the effect and risks of evobrutinib in patients with multiple sclerosis. (Funded by EMD Serono; ClinicalTrials.gov number, NCT02975349.).
Evobrutinib, a covalent Bruton's tyrosine kinase inhibitor: Mass balance, elimination route, and metabolism in healthy participants
Clin Transl Sci2021 Nov;14(6):2420-2430.PMID: 34374206DOI: 10.1111/cts.13108
The highly selective, covalent Bruton's tyrosine kinase inhibitor evobrutinib is under investigation for treatment of patients with multiple sclerosis (MS). Early clinical studies in healthy participants and patients with relapsing MS indicated that evobrutinib is well-tolerated and effective. We undertook a mass balance study in six men who received a single 75-mg oral dose of evobrutinib containing ~ 3.6 MBq (100 μCi) 14 C-evobrutinib, to determine the absorption, metabolic pathways, and routes of excretion of evobrutinib. The primary objectives of this phase I study (NCT03725072) were to (1) determine the rates and routes of total radioactivity excretion, including the mass balance of total drug-related radioactivity in urine and feces, (2) assess the pharmacokinetics (PKs) of total radioactivity in blood and plasma, and (3) characterize the plasma PKs of evobrutinib. Exploratory end points included identifying and quantifying evobrutinib and its metabolites in plasma and excreta (urine and feces) and exploring key biotransformation pathways and clearance mechanisms. Evobrutinib was primarily eliminated in feces (arithmetic mean percentage, SD, 71.0, 2.1) and, to a lesser extent, in urine (20.6, 2.0), with most of the total radioactivity (85.3%) excreted in the first 72 h after administration. No unchanged evobrutinib was detected in excreta. Evobrutinib was rapidly absorbed and substantially metabolized upon absorption. Only one major metabolite M463-2 (MSC2430422) was identified in plasma above the 10% of total drug exposure threshold, which classifies M463-2 (MSC2430422) as a major metabolite according to the US Food and Drug Administration (FDA; metabolites in safety testing [MIST]) and the European Medicines Agency (EMA; International Conference on Harmonization [ICH] M3). These results support further development of evobrutinib and may help inform subsequent investigations.
Comparative Analysis of BTK Inhibitors and Mechanisms Underlying Adverse Effects
Front Cell Dev Biol2021 Mar 11;9:630942.PMID: 33777941DOI: 10.3389/fcell.2021.630942
The cytoplasmic protein-tyrosine kinase BTK plays an essential role for differentiation and survival of B-lineage cells and, hence, represents a suitable drug target. The number of BTK inhibitors (BTKis) in the clinic has increased considerably and currently amounts to at least 22. First-in-class was ibrutinib, an irreversible binder forming a covalent bond to a cysteine in the catalytic region of the kinase, for which we have identified 228 active trials listed at ClinicalTrials.gov. Next-generation inhibitors, acalabrutinib and zanubrutinib, are approved both in the United States and in Europe, and zanubrutinib also in China, while tirabrutinib is currently only registered in Japan. In most cases, these compounds have been used for the treatment of B-lymphocyte tumors. However, an increasing number of trials instead addresses autoimmunity and inflammation in multiple sclerosis, rheumatoid arthritis, pemphigus and systemic lupus erythematosus with the use of either irreversibly binding inhibitors, e.g., evobrutinib and tolebrutinib, or reversibly binding inhibitors, like fenebrutinib. Adverse effects (AEs) have predominantly implicated inhibition of other kinases with a BTKi-binding cysteine in their catalytic domain. Analysis of the reported AEs suggests that ibrutinib-associated atrial fibrillation is caused by binding to ERBB2/HER2 and ERBB4/HER4. However, the binding pattern of BTKis to various additional kinases does not correlate with the common assumption that skin manifestations and diarrhoeas are off-target effects related to EGF receptor inhibition. Moreover, dermatological toxicities, diarrhoea, bleedings and invasive fungal infections often develop early after BTKi treatment initiation and subsequently subside. Conversely, cardiovascular AEs, like hypertension and various forms of heart disease, often persist.