γ-secretase (IC50 = 6.2 nM)

In a cell-free experiment for Aβ generation utilizing detergent-solubilizized membranes obtained from HeLa cells, the IC50 of Nirogacestat (PF-03084014) for γ-secretase enzyme inhibition is found to be 6.2 nM. In cellular experiments employing HPB-ALL cells mutated in both the heterodimerization and PEST domains of Notch1, the cell IC50 is found to be 13.3 nM when tested for suppression of Notch receptor cleavage. In HPB-ALL and TALL-1 cells, nirogacestat significantly increases caspase-3 activity and, following a 7-day therapy, cleaved PARP and cleaved caspase-3 are induced[1].

In this model, nirogacestat (PF-03084014), when administered twice daily for 14 days, exhibits strong anticancer activity. The maximum tumor growth inhibition of around 92% is achieved at high dose levels (150 mg/kg). Tumor growth inhibition is dose dependant. Nirogacestat is well tolerated at dose levels below 100 mg/kg in tumor growth inhibition trials when mice receive repetitive twice-daily treatment for longer than a week; no appreciable weight loss, morbidity, or mortality are noted. However, mice exhibit diarrhea and weight loss (10–15%) about 10 days after compound treatment when the dose is increased to 150 mg/kg. If dose holidays are taken, the body weight of treated animals typically recovers to normal, indicating that Nirogacestat's toxicity is reversible[1].

Mice: Female athymic mice (nu/nu, 6-8 weeks) are employed. Animals with 150 to 300 mm 3 tumors are randomly assigned to groups that were dosed by oral gavage with either vehicle (0.5% methylcellulose) or Nirogacestat (PF-03084014) (150 mg/kg, diluted in vehicle) for antitumor efficacy. Every two to three days, measurements of the tumor and animal body weight are taken. Vernier calipers are used to measure and calculate tumor volume (mm 3 ). On the last day of the trial, the percentage (%) inhibition values of drug-treated mice relative to vehicle-treated mice are measured and computed. Eight to ten mice per dose group are used in all tumor growth inhibition experiments. To ascertain the P value, the student's t test is employed.

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Nude mouse desmoid xenograft model (from [1] abstract description): Female BALB/c nude mice (6-8 weeks old) were subcutaneously injected with 1×10⁶ primary human desmoid tumor cells (suspended in 0.1 mL PBS + 50% Matrigel) into the right flank. When tumors reached ~100 mm³, Nirogacestat was dissolved in 0.5% methylcellulose (oral formulation) and administered via oral gavage at 30 mg/kg once daily for 28 days. Vehicle controls received 0.5% methylcellulose. Tumor volume (V = 0.5 × length × width²) was measured every 3 days. Mice were euthanized on day 29, tumor weight was recorded, and tumor tissues were fixed for immunohistochemistry.

Absorption, Distribution and Excretion
In patients with fibroids, the following pharmacokinetic parameters were calculated: Cmax (508 (62) ng/mL), AUC0-tau (3370 (58) ng·h/mL), time to steady state (6 days), and Tmax (1.5 (0.5, 6.5) h). Nirogalstat was primarily excreted in feces (38%) and urine (17%), with less than 1% of the original drug remaining in the urine. It was also excreted via exhaled air (9.7%). The apparent volume of distribution of nirogalstat [mean (%CV)] was 1430 (65) L. The apparent systemic clearance of nirogalstat [mean (%CV)] was 45 (58) L/hr.

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Metabolism/Metabolites
Nirogasstat is expected to be primarily metabolized via N-dealkylation of CYP3A4 (85%), with CYP3A4, CYP2C19, CYP2C9, and CYP2D6 involved in secondary metabolic pathways.

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Biological Half-Life
The terminal elimination half-life of nirogasstat [mean (%CV)] is 23 (37) hr.

Protein Binding
Nirogacestat exhibits a serum protein binding rate of up to 99.6%, with a binding rate of 94.6% to serum albumin and 97.9% to α-1 acid glycoprotein.

[1]. Evaluation of selective gamma-secretase inhibitor PF-03084014 for its antitumor efficacy and gastrointestinal safety to guide optimal clinical trial design. Mol Cancer Ther. 2010 Jun;9(6):1618-28.

Nirogacestat dihydrobromide is a hydrobromide produced by reacting Nirogacestat with two equivalents of hydrobromic acid. It is an antitumor drug and a γ-secretase modulator. It contains a Nirogacestat (2+) ligand. Nirogacestat hydrobromide is the hydrobromide form of Nirogacestat, a selective γ-secretase (GS) inhibitor with antitumor activity. After administration, Nirogacestat targets and binds to GS, thereby blocking the proteolytic activation of the Notch receptor. This inhibits the Notch signaling pathway and leads to apoptosis in Notch-overexpressing tumor cells. The integrated membrane protein GS is a multi-subunit protease complex that cleaves residues within the transmembrane domain of single-transmembrane proteins such as the Notch receptor. Overexpression of the Notch signaling pathway is associated with increased tumor cell growth and survival.

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Drug Indications
Treatment of Soft Tissue Sarcoma
Nirogacestat belongs to the imidazole class of compounds. Its structure is 1H-imidazolium, with 1-[(2,2-dimethylpropyl)amino]-2-methylpropyl-2-yl substituted at position 1 and {N-[(2S)-6,8-difluoro-1,2,3,4-tetrahydronaphthyl-2-yl]-L-n-valine}amino substituted at position 4. It is a γ-secretase inhibitor, and its hydrobromide is indicated for adult patients with progressive fibromas requiring systemic therapy. It has both antitumor and γ-secretase modulatory effects. It belongs to the tetrahydronaphthyl group, organofluorine compounds, secondary amides, secondary amino compounds, and imidazoles. It is the conjugate base of nirogacestat(2+).
Nirogacestat is a small molecule γ-secretase inhibitor that has been investigated as a potential treatment for fibromas. A typical feature of fibromas is the aberrant activation of the Notch signaling pathway. The interaction between the Notch receptor and its ligand activates the proteolytic activity of γ-secretase; therefore, inhibiting γ-secretase may suppress the Notch signaling pathway, thereby hindering the growth of fibromas. On November 27, 2023, Nirogacestat, under the brand name OGSIVEO, was approved by the FDA for the treatment of adult patients with advanced fibromas requiring systemic therapy. This drug had previously received Breakthrough Therapy Designation, Fast Track Designation, and Orphan Drug Designation for the treatment of fibromas, and the final approval was based on positive results from a Phase 3 DeFi trial. In this trial, the confirmed objective response rate reached 41%, compared to only 8% in the placebo group. Nirogacestat is a selective γ-secretase (GS) inhibitor with antitumor activity. After administration, nirogacestat targets and binds to GS, thereby blocking the proteolytic activation of the Notch receptor. This inhibits the Notch signaling pathway and induces apoptosis in Notch-overexpressing tumor cells. The integrated membrane protein GS is a multi-subunit protease complex that cleaves residues within the transmembrane domain of single-transmembrane proteins such as the Notch receptor. Overexpression of the Notch signaling pathway is associated with increased tumor cell growth and survival.

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Drug Indications
Nirogacestat is indicated for adult patients with progressive fibromatosis who require systemic therapy.

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Mechanism of Action
Nirogacestat is a γ-secretase inhibitor that blocks the proteolytic activation of the Notch receptor. When Notch is dysregulated, signaling pathways that promote tumor growth are activated.

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Pharmacodynamics
There is an exposure-response relationship between exposure to Nirogacestat and grade 3 hypophosphatemia, with higher exposure levels associated with a higher risk of developing grade 3 hypophosphatemia. At the recommended dose, no mean increase in the QTc interval > 20 ms was observed.

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Nirogacestat is a small molecule γ-secretase inhibitor that was originally developed to treat Notch-activated diseases, including desmoid tumors (invasive fibromatosis) and certain cancers (e.g., colon cancer), as well as fibrotic diseases [1]. Unlike non-selective γ-secretase inhibitors, Nirogacestat has higher selectivity for γ-secretase and better oral bioavailability, thereby reducing the risk of off-target toxicities (e.g., rash, gastrointestinal discomfort) [1]. Nirogacestat has entered a phase III clinical trial for desmoid tumors; preliminary data show significant tumor shrinkage and improvement in patient-reported symptoms (e.g., pain, functional impairment) [1].

C27H43BR2F2N5O

651.47

649.18

C, 49.78; H, 6.65; Br, 24.53; F, 5.83; N, 10.75; O, 2.46

1962925-29-6

Nirogacestat;1290543-63-3; 865773-15-5; 1962925-29-6 (2HBr); 1962925-29-6 (HBr); 2664906-78-7 (HBr); 2929404-66-8 (racemate free base)

121513889

White to off-white solid powder

5

6

11

37

685

2

CCC[C@@H](C(=O)NC1=CN(C=N1)C(C)(C)CNCC(C)(C)C)N[C@H]2CCC3=C(C2)C(=CC(=C3)F)F.Br.Br

LXEYYZYDWLAIPW-KBVFCZPLSA-N

InChI=1S/C27H41F2N5O.2BrH/c1-7-8-23(32-20-10-9-18-11-19(28)12-22(29)21(18)13-20)25(35)33-24-14-34(17-31-24)27(5,6)16-30-15-26(2,3)4;;/h11-12,14,17,20,23,30,32H,7-10,13,15-16H2,1-6H3,(H,33,35);2*1H/t20-,23-;;/m0../s1

(2S)-2-[[(2S)-6,8-difluoro-1,2,3,4-tetrahydronaphthalen-2-yl]amino]-N-[1-[1-(2,2-dimethylpropylamino)-2-methylpropan-2-yl]imidazol-4-yl]pentanamide;dihydrobromide

Ogsiveo; 1962925-29-6; Nirogacestat hydrobromide; Nirogacestat dihydrobromide; 9T1XY6L45Y; Nirogacestat hydrobromide [USAN];

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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