Cytochrome P450 3A4; Organic Anion Transporting Polypeptide 1B1; Organic Anion Transporting Polypeptide 1B3; Neuroprotective agent

The mechanism of action of trofinetide is as a Cytochrome P450 3A4 Inhibitor, and Organic Anion Transporting Polypeptide 1B1 Inhibitor, and Organic Anion Transporting Polypeptide 1B3 Inhibitor.

Trofinetide (NNZ-2566), as opposed to tax substance therapy, promoted cytokine activation three days following PBBI in inhibitory ballistic-like brain damage (PBBI). Treatment with trafeneptide for 12 hours after PBBI resulted in a considerable decrease in the amount of mRNA for IL-6 (79%), E-selectin (81%), IL-1β (76%), and TNF-α (72%), in the injured hemisphere. levels rose, and between 12 and 24 hours was when the maximum inhibition happened. Treatment with trofinetide did not influence the upregulation of IL-6 expression induced by PBBI at any time point, but it dramatically decreased the upregulation of IL-1β, INF-γ, and TNF-α expression caused by injury. IL-1β expression in the cerebral hemispheres was decreased by a peptide administered with tramenide for a prolonged period of 7 days following PBBI [1]. Two hours after temporary middle cerebral artery blockage (pMCAo), nonconvulsive reflux (NCS) was lessened by high doses of trofinetide (NNZ-2566) (10 and 100 mg/kg bolus followed by continuous infusion). Trofinetide, in comparison to animals administered with a vehicle, totally reduced the delayed onset of NCS at all dosages [2].

Absorption, Distribution and Excretion
Systemic exposure to trofinetide is dose-dependent within the studied dose range. The time to peak exposure (Tmax) is approximately 2 to 3 hours after administration. According to mass balance studies, at least 84% of the orally administered 12,000 mg trofinetide is absorbed. High-fat meals have a negligible effect on drug exposure, reducing Cmax by only about 20%. Trofinetide is primarily excreted in the urine, with a small amount excreted in the feces. Approximately 80% of the dose recovered in the urine is in its unchanged parent form. Following oral administration, the apparent volume of distribution of trofinetide in healthy adult subjects is approximately 80 liters. No drug accumulation was observed after multiple administrations. No further information is available. Metabolism/Metabolites Trofinetide is primarily metabolized by CYP450 enzymes; hepatic metabolism is not its primary clearance pathway.

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Biological Half-Life
The effective elimination half-life of trafenidone in healthy subjects after oral administration is approximately 1.5 hours.

Effects During Pregnancy and Lactation
◉ Overview of Use During Lactation
There is currently no information regarding the use of trafeneptide during lactation. If the mother requires trafeneptide, breastfeeding should not be discontinued. The infant's diarrhea and weight gain should be closely monitored while trafeneptide is used until more data are available.
◉ Effects on Breastfed Infants
No published information found as of the revision date.
◉ Effects on Lactation and Breast Milk
No published information found as of the revision date.

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Protein Binding
Protein binding to human plasma is less than 6%.

[1]. NNZ-2566 treatment inhibits neuroinflammation and pro-inflammatory cytokine expression induced by experimental penetrating ballistic-like brain injury in rats. J Neuroinflammation. 2009 Aug 5;6:19.
[2]. NNZ-2566, a glypromate analog, attenuates brain ischemia-induced non-convulsive seizures in rats. J Cereb Blood Flow Metab. 2009 Dec;29(12):1924-32.
[3]. Cartagena CM, Phillips KL, Williams GL, et al. Mechanism of action for NNZ-2566 anti-inflammatory effects following PBBI involves upregulation of immunomodulator ATF3. Neuromolecular Med. 2013;15(3):504-514

Trofinetide is a tripeptide composed of glycine, 2-methyl-L-proline, and L-glutamate linked by peptide bonds. It is currently the only FDA-approved drug specifically for the treatment of Rett syndrome in adults and children aged two years and older. It has neuroprotective and anti-inflammatory effects. Trofinetide is a tripeptide whose structure includes pyrrolidine carboxamide, a primary amino compound, a secondary carboxamide, a tertiary carboxamide, and a dicarboxylic acid. Functionally, it is associated with L-glutamate and glycine. Trofinetide is a novel synthetic analog of glycine-proline-glutamate (GPE), a naturally occurring protein in the brain and the N-terminal tripeptide of insulin-like growth factor 1 (IGF-1). Trofinetide received FDA approval on March 10, 2023, for the treatment of Rett syndrome, an X-linked neurodevelopmental disorder characterized by a range of cognitive, motor, and autonomic symptoms. Trofinetide is believed to work by reducing neuronal inflammation and apoptosis.
The mechanism of action of tofenitide is as an inhibitor of cytochrome P450 3A4, organic anion transport peptide 1B1, and organic anion transport peptide 1B3.
Pharmaceutical Indications

Tofenitide is indicated for the treatment of Rett syndrome in adults and children aged two years and older.
Mechanism of Action

Most cases of Rett syndrome are associated with loss-of-function mutations in the gene encoding methyl CpG-binding protein 2 (MECP2). MECP2 is a DNA-binding protein that plays a role in the epigenetic regulation of gene expression. These mutations are thought to lead to immature cortical synaptic development, abnormal brain cholesterol metabolism, and consequently, abnormal neuronal development and signal transduction. Rett syndrome is more common in girls than boys. The exact mechanism of action of trefenide in treating Rett syndrome is not fully elucidated. In mouse studies, GPE improved motor and cardiopulmonary function, increased brain weight, and prolonged lifespan in MECP2-deficient mice. As a GPE analog, trofinetide reduced apoptosis and infarct size in a dose-dependent manner in a rat model of hypoxic injury. Trofinetide exerts multiple effects, reducing tissue damage and apoptosis caused by inflammation and excitotoxicity, thereby protecting neurons and surrounding structures.

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Pharmacodynamics
Trofinetide can alleviate and relieve symptoms of Rett syndrome. It is an analog of glycine-proline-glutamate (GPE), a naturally occurring protein in the brain and the N-terminal tripeptide of insulin-like growth factor 1 (IGF-1). GPE has neuroprotective effects: it can protect neurons from glutamate-mediated excitotoxicity and oxidative stress damage at nanomolar concentrations. Compared to GPE, trofinetide, with its longer half-life, is considered to have similar neuroprotective effects.

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Trofinetide is used to alleviate and relieve symptoms of Rett syndrome. It is an analogue of glycine-proline-glutamate (GPE), a naturally occurring protein in the brain and the N-terminal tripeptide of insulin-like growth factor 1 (IGF-1). GPE has neuroprotective effects: it can protect neurons from glutamate-mediated excitotoxicity and oxidative stress damage at nanomolar concentrations. Trofinetide, with its longer half-life compared to GPE, is thought to have similar neuroprotective effects.

C13H21N3O6

315.322343587875

315.143

C, 49.52; H, 6.71; N, 13.33; O, 30.44

853400-76-7

11318905

H-Gly-aMePro-Glu-OH; glycyl-alpha-methyl-L-prolyl-L-glutamic acid; Gly-{Pro(α-Me)}-Glu

GXE; G{Pro(α-Me)}E

White to off-white solid powder

1.4±0.1 g/cm3

655.4±55.0 °C at 760 mmHg

350.2±31.5 °C

0.0±4.3 mmHg at 25°C

1.558

-2.22

4

7

7

22

480

2

C([C@@]1(CCCN1C(=O)CN)C)(=O)N[C@H](C(=O)O)CCC(=O)O

BUSXWGRAOZQTEY-SDBXPKJASA-N

InChI=1S/C13H21N3O6/c1-13(5-2-6-16(13)9(17)7-14)12(22)15-8(11(20)21)3-4-10(18)19/h8H,2-7,14H2,1H3,(H,15,22)(H,18,19)(H,20,21)/t8-,13-/m0/s1

((S)-1-glycyl-2-methylpyrrolidine-2-carbonyl)-L-glutamic acid

NNZ-2566; Daybue; NNZ2566; Trofinetide; 853400-76-7; DAYBUE; L-Glutamic acid, glycyl-2-methyl-L-prolyl-; NNZ 2566

2934.99.9001

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 and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~110 mg/mL (~348.85 mM)
H2O : ≥ 50 mg/mL (~158.57 mM)
DMSO : ~25 mg/mL (~79.28 mM)

Solubility in Formulation 1: 100 mg/mL (317.14 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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