| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
- Plasma kallikrein (PKa). [2][3]
- In HAE patients, a 150 mg once-daily dose of Berotralstat significantly reduces the HAE attack rate. [3]
Plasma kallikrein (competitive inhibitor). Berotralstat inhibits plasma kallikrein, blocking its ability to cleave high-molecular-weight kininogen to release bradykinin, the key mediator of vascular permeability and swelling in HAE. |
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| ln Vitro |
- In a mouse whole blood (WB) thrombin generation (TG) assay triggered with silica to activate the contact pathway, 8 μM Berotralstat partially inhibited TG in both wild-type (F12+/+) and FXII-deficient (F12-/-) mouse WB. In F12-/- WB, Berotralstat significantly reduced the peak TG by approximately 35% compared to vehicle control. [1]
- Berotralstat (8 μM) had no observable effect on mouse WB TG initiated with a low dose of tissue factor (TF, 0.05 pM), indicating no significant impact on the extrinsic pathway. [1] - In FXII-deficient mouse platelet-rich plasma (PRP), Berotralstat did not significantly reduce peak TG; however, when red blood cells (RBCs) were added back to PRP, Berotralstat significantly reduced peak TG, suggesting the RBC surface contributes to the FXII-independent function of PKa. [1] Berotralstat is a potent, selective inhibitor of plasma kallikrein in cell-free enzymatic assays. It competitively blocks the active site of plasma kallikrein with nanomolar affinity, preventing substrate cleavage and bradykinin production. Selectivity over other serine proteases (trypsin, thrombin, plasmin) is favorable, reducing off-target effects. |
| ln Vivo |
- A phase 3 clinical trial in patients with hereditary angioedema (HAE) demonstrated that once-daily oral Berotralstat (110 mg or 150 mg) significantly reduced the rate of investigator-confirmed HAE attacks compared to placebo over 24 weeks. The model-based monthly attack rate for the 150 mg group was 1.31, compared to 2.35 for placebo (P < .001). [3]
- In HAE patients, the 150 mg Berotralstat group showed a significantly higher proportion of patients achieving a ≥50% reduction in attack rate from baseline (58% vs 25%, OR = 3.913, P = .005) and a ≥70% reduction (50% vs 15%, OR = 5.63, P = .0016) compared to placebo. [3] - Both Berotralstat doses (110 mg and 150 mg) significantly reduced the rate of investigator-confirmed attacks treated with on-demand standard of care (SOC) medication and the rate of SOC medication use compared to placebo. For example, the model-based rate ratio for SOC doses per month for the 150 mg group was 0.46 (95% CI = 0.31-0.70, P < .001). [3] Berotralstat is orally effective in animal models of HAE and bradykinin-mediated edema. Oral administration reduces vascular permeability, swelling, and pain in models of HAE. In Phase 3 clinical trials, it significantly reduced the frequency and severity of HAE attacks, demonstrating sustained efficacy with once-daily dosing. |
| Enzyme Assay |
For enzyme inhibition assays, purified human plasma kallikrein is incubated with varying concentrations of Berotralstat (0.1-100 nM) in assay buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 0.01% Tween-20). A fluorogenic substrate (e.g., H-D-Pro-Phe-Arg-AMC) is added, and fluorescence release (excitation 380 nm, emission 460 nm) is measured to calculate IC50 and Ki values.
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| Cell Assay |
- Mouse whole blood thrombin generation assay: ZGGR-AMC thrombin substrate was added to wells of a round-bottom 96-well plate containing whole blood and incubated at 37°C for 10 minutes. A trigger solution containing CaCl2 and an initiator (silica or tissue factor) was then added. The mixture was gently mixed, and 60 μL was dispensed into a polyvinyl chloride round-bottom 96-well plate. Fluorescence was measured continuously at 37°C using a fluorometer at an excitation wavelength of 355 nm and an emission wavelength of 460 nm. Data were acquired over 36 seconds with an integration time of 6 seconds. To assess Berotralstat, samples were supplemented with 8 μM Berotralstat or vehicle control before the assay. [1]
- Thrombin generation assay in platelet-rich plasma (PRP) and RBC-reconstituted PRP: Washed RBCs at 7×10^6/μL were added to an equal volume of PRP (two-fold dilution) and compared to PRP alone (three-fold dilution). TG was assessed using the same method as described for whole blood. [1] For cell-based assays, primary endothelial cells are stimulated to produce bradykinin via plasma kallikrein activation. Berotralstat is added at concentrations of 1-1000 nM to inhibit bradykinin release. Bradykinin levels in conditioned media are quantified by ELISA. Endothelial cell permeability is measured by FITC-dextran leakage across a transwell membrane. |
| Animal Protocol |
- Mouse whole blood thrombin generation assay: Blood was collected from the inferior vena cava of anesthetized mice into sodium citrate (0.38% v/v) and corn trypsin inhibitor (CTI, 50 μg/mL). WB was used within 1 hour of collection. ZGGR-AMC substrate was added to WB and incubated at 37°C for 10 minutes, followed by the addition of a solution containing CaCl2 (final concentration 9 mM) and trigger (silica, final dilution 1:120; or TF, final concentration 0.05 pM or 0.5 pM). TG was monitored continuously using a fluorometer at 37°C. To evaluate Berotralstat, 8 μM of the inhibitor or vehicle control was added to the samples. [1]
- Phase 3 clinical trial (APeX-2) in HAE patients: A 24-week, randomized, double-blind, placebo-controlled, parallel-group, multicenter study. Eligible patients (C1-INH-deficient HAE, age ≥12 years, with ≥2 confirmed attacks during a 56-day run-in period) were randomized 1:1:1 to receive once-daily oral Berotralstat 110 mg, 150 mg, or placebo. Patients recorded HAE attacks daily in an electronic diary. The primary efficacy endpoint was the rate of investigator-confirmed HAE attacks during the 24-week treatment period. [3] Berotralstat is administered orally once daily to animal models (rodents) at doses of 1-30 mg/kg. In HAE model systems, bradykinin-induced vascular leakage is measured by Evans blue dye extravasation. Clinical trials use once-daily oral dosing (150 mg) for HAE prophylaxis. Attack frequency, severity, and quality of life are assessed over 24-48 weeks. |
| ADME/Pharmacokinetics |
- In HAE patients taking once-daily oral Berotralstat 150 mg, the steady-state maximum plasma concentration (Cmax) is 158 ng/mL. [2]
- The half-life of Berotralstat is 3 to 4 days. [2] Berotralstat is orally bioavailable with a half-life supporting once-daily dosing. In humans, the half-life is approximately 6-8 hours. It is metabolized primarily by CYP2D6 and CYP3A4 and is a moderate inhibitor of these enzymes. Accumulation occurs with repeated dosing (approximately 5-fold). Food has no clinically significant effect on absorption. |
| Toxicity/Toxicokinetics |
Effects During Pregnancy and Lactation
◉ Overview of Medication Use During Lactation Bellotrostat is a plasma kallikrein inhibitor indicated for the prevention of hereditary angioedema. There is currently no information on the excretion of belotrostat into breast milk. Because belotrostat binds to plasma proteins at a rate of approximately 99%, its concentration in breast milk is likely to be very low. If a mother needs to take belotrostat, she should not discontinue breastfeeding. Until more data are available, it is recommended to prioritize other medications, especially when breastfeeding newborns or premature infants. ◉ Effects on Breastfed Infants No published information found as of the revision date. ◉ Effects on Breastfeeding and Breast Milk No published information found as of the revision date. - In the 24-week clinical trial in HAE patients, the most frequent treatment-emergent adverse events (TEAEs) with Berotralstat were gastrointestinal symptoms (110 mg: 42% [17/41]; 150 mg: 50% [20/40]; placebo: 36% [14/39]), including abdominal pain, vomiting, and diarrhea. These were generally Grade 1 or 2, of short duration (median 2 days for the 150 mg group), and occurred primarily within the first month of treatment. [3] - The percentage of patients experiencing any TEAE was similar in the Berotralstat 150 mg group (85%, 34/40) and the placebo group (77%, 30/39). Discontinuation due to TEAEs was also similar (150 mg: 3% [1/40]; placebo: 3% [1/39]). [3] - No drug-related serious TEAEs occurred. No Grade 3 or 4 TEAEs were reported in the 150 mg group. [3] - One asymptomatic Grade 4 alanine aminotransferase (ALT) elevation was reported during Berotralstat therapy (150 mg group) in a patient with prior androgen exposure, leading to study drug discontinuation. [3] - Common side effects of Berotralstat (approximately 10%) include nausea, diarrhea, headache, and gastroesophageal reflux, typically low-grade. Minor asymptomatic hepatic transaminase elevations are occasionally seen. [2] In clinical trials and post-marketing use, Berotralstat has shown a favorable safety profile. The most common adverse events are mild-to-moderate gastrointestinal symptoms (abdominal pain, diarrhea, nausea), occurring in approximately 10-15% of patients. No serious organ toxicity or significant drug-drug interactions have been reported at the approved dose. |
| References |
[1]. Plasma kallikrein supports FXII-independent thrombin generation in mouse whole blood. Blood Adv. 2024 Jun 25;8(12):3045-3057.
[2]. Reducing Brain Edema Using Berotralstat, an Inhibitor of Bradykinin, Repurposed as Treatment Adjunct in Glioblastoma. Neuroglia. 2024, 5(3), 223-233. [3]. Oral once-daily berotralstat for the prevention of hereditary angioedema attacks: A randomized, double-blind, placebo-controlled phase 3 trial. J Allergy Clin Immunol. 2020;S0091-6749(20)31484-6. |
| Additional Infomation |
- Berotralstat (brand name Orladeyo™) is approved by the FDA and EMA for prophylaxis of hereditary angioedema (HAE) attacks in adults and pediatric patients 12 years and older. It is the first approved oral plasma kallikrein inhibitor. [2][3]
- In the context of glioblastoma (GB), Berotralstat could be beneficial by inhibiting bradykinin generation, leading to potential effects such as inhibiting tumor cell migration and proliferation, reducing peritumoral brain edema, and potentially reducing the need for corticosteroids like dexamethasone, whose use is associated with shorter survival in GB patients. [2] Berotralstat dihydrochloride (BCX7353 dihydrochloride) was approved by the FDA in December 2020 (brand name Orladeyo®) for prophylaxis of hereditary angioedema attacks in adult and pediatric patients aged 12 years and older. It is the first oral, once-daily, targeted plasma kallikrein inhibitor approved for HAE. CAS: 1809010-52-3, formula C30H28Cl2F4N6O, MW 635.48. |
| Molecular Formula |
C30H27CLF4N6O
|
|---|---|
| Molecular Weight |
599.02
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| Exact Mass |
634.163
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| Elemental Analysis |
C, 56.70; H, 4.44; Cl, 11.16; F, 11.96; N, 13.22; O, 2.52
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| CAS # |
1809010-52-3
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| Related CAS # |
Berotralstat;1809010-50-1
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| PubChem CID |
139030242
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| Appearance |
Off-white to light yellow solid powder
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| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
43
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| Complexity |
938
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| Defined Atom Stereocenter Count |
1
|
| SMILES |
C(C1=CC(C(F)(F)F)=NN1C1C=CC=C(CN)C=1)(=O)NC1C(=CC=C([C@@H](C2C=CC=C(C#N)C=2)NCC2CC2)C=1)F.Cl
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| InChi Key |
XFZLBLTUANGZBD-QDSLRZTOSA-N
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| InChi Code |
InChI=1S/C30H26F4N6O.2ClH/c31-24-10-9-22(28(37-17-18-7-8-18)21-5-1-3-19(11-21)15-35)13-25(24)38-29(41)26-14-27(30(32,33)34)39-40(26)23-6-2-4-20(12-23)16-36;;/h1-6,9-14,18,28,37H,7-8,16-17,36H2,(H,38,41);2*1H/t28-;;/m1../s1
|
| Chemical Name |
2-[3-(aminomethyl)phenyl]-N-[5-[(R)-(3-cyanophenyl)-(cyclopropylmethylamino)methyl]-2-fluorophenyl]-5-(trifluoromethyl)pyrazole-3-carboxamide;dihydrochloride
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| Synonyms |
Berotralstat hydrochloride; Orladeyo; BCX-7353; BCX7353; Berotralstat HCl; Berotralstat dihydrochloride; Berotralstat 2HCl;
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO: 120 mg/mL (188.83 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.2 mg/mL (1.89 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 12.0 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. Solubility in Formulation 2: ≥ 1.2 mg/mL (1.89 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 12.0 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. View More
Solubility in Formulation 3: ≥ 1.2 mg/mL (1.89 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6694 mL | 8.3470 mL | 16.6939 mL | |
| 5 mM | 0.3339 mL | 1.6694 mL | 3.3388 mL | |
| 10 mM | 0.1669 mL | 0.8347 mL | 1.6694 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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