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 was dose-proportional across the studied dose range. The Tmax is about two to three hours after administration. Based on the mass balance study, at least 84% of the administered oral dose of 12,000 mg trofinetide was absorbed. A high-fat meal had negligible effects on drug exposure and reduced the Cmax by approximately 20%.
Trofinetide is primarily excreted in urine, with minor excretion in feces. Approximately 80% of the dose recovered in urine was in the unchanged parent drug form.
Following oral administration, the apparent volume of distribution of trofinetide in healthy adult subjects was approximately 80 L. Minimal to no accumulation was observed following multiple-dose administration.
No information is available.

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Metabolism / Metabolites
Trofinetide is not significantly metabolized by CYP450 enzymes. Hepatic metabolism is not a significant route of trofinetide elimination.

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Biological Half-Life
The effective elimination half-life of orally administered trofinetide in healthy subjects is about 1.5 hours.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the use of trofinetide during breastfeeding. If trofinetide is required by the mother, it is not a reason to discontinue breastfeeding. Until more data are available, trofinetide should be used with careful infant monitoring for diarrhea and proper weight gain.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

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

[1]. Wei HH, et al. 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]. Lu XC, et al. 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 comprising of glycine, 2-methyl-L-proline, and L-glutamic acid joined in sequence by peptide linkages. It is the first and only treatment approved by the FDA specifically indicated for Rett syndrome in adults and pediatric patients two years of age and older. It has a role as a neuroprotective agent and an anti-inflammatory agent. It is a tripeptide, a pyrrolidinecarboxamide, a primary amino compound, a secondary carboxamide, a tertiary carboxamide and a dicarboxylic acid. It is functionally related to a L-glutamic acid and a glycine.
Trofinetide is a novel synthetic analog of [glypromate], also known as 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 was approved by the FDA on March 10, 2023, for the treatment of Rett syndrome, which is an X-linked neurodevelopmental disorder characterized by a range of cognitive, motor, and autonomic symptoms. Trofinetide is believed to work by reducing inflammation and apoptosis of neurons.
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.

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Drug Indication
Trofinetide is indicated for the treatment of Rett syndrome in adults and pediatric patients two years of age and older.

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Mechanism of Action
Most cases of Rett syndrome are associated with loss-of-function mutations in a gene encoding methyl CpG binding protein 2 (MECP2), a DNA binding protein with a role in epigenetic regulation of gene expression. These mutations are believed to lead to synaptic immaturation in the cortex, aberrant metabolism of brain cholesterol resulting in abnormal neuronal development, and abnormal neuronal signaling. Rett syndrome more commonly occurs in girls than boys. The exact mechanism of action of trofinetide in Rett syndrome has not been fully elucidated. In mice studies, GPE improved motor and cardiorespiratory function, increased brain weight, and prolonged the lifespan in MECP2-deficient mice. As a GPE analog, trofinetide similarly attenuated apoptosis and reduced infarct size in a dose-dependent manner in a rat model of hypoxic insult. Trofinetide exerts a multi-faceted action to reduce inflammation, excitotoxicity-induced tissue damage, and apoptosis, thereby protecting the neurons and their surrounding infrastructure.

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Pharmacodynamics
Trofinetide works to reduce and alleviate the symptoms of Rett syndrome. It is an analog of [glypromate] or 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 exhibits neuroprotective properties: it protects neurons from glutamate-mediated excitotoxicity and oxidative stress at nanomolar concentrations. With a longer half-life than GPE, trofinetide is believed to exhibit similar neuroprotective properties.

315.143

853400-76-7

11318905

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](CCC(=O)O)C(=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 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.)