Fmoc-PEA (Example 1-2) can be used as a cleavable (degradable) cross-linker for Antibody-drug conjugates (ADC).

Physicochemical Properties

Molecular Formula	C17H18NO6P
Molecular Weight	363.3017
Exact Mass	363.087
CAS #	329223-23-6
PubChem CID	11187609
Appearance	Off-white to light yellow solid powder
LogP	1.3
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	7
Heavy Atom Count	25
Complexity	487
Defined Atom Stereocenter Count	0
SMILES	P(=O)(O[H])(O[H])OC([H])([H])C([H])([H])N([H])C(=O)OC([H])([H])C1([H])C2=C([H])C([H])=C([H])C([H])=C2C2=C([H])C([H])=C([H])C([H])=C12
InChi Key	YHUHBHVNOVAEMP-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H18NO6P/c19-17(18-9-10-24-25(20,21)22)23-11-16-14-7-3-1-5-12(14)13-6-2-4-8-15(13)16/h1-8,16H,9-11H2,(H,18,19)(H2,20,21,22)
Chemical Name	9H-fluoren-9-ylmethyl N-(2-phosphonooxyethyl)carbamate
HS Tariff Code	2934.99.9001
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 and light.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro	- Fmoc-PEA functions as a phosphate-based linker to construct drug conjugates; the conjugates prepared with Fmoc-PEA achieve intracellular delivery of active drugs in vitro cell experiments [1] - The phosphate ester bond of Fmoc-PEA is hydrolyzed by intracellular phosphatases, enabling controlled release of the conjugated active drug inside target cells, which maintains the biological activity of the drug against target cells [1]
Cell Assay	- Cellular Uptake Assay: Fluorescently labeled drug conjugates constructed with Fmoc-PEA were co-incubated with target cells for different time periods [1] After incubation, cells were washed to remove unbound conjugates, and intracellular fluorescence intensity was detected by fluorescence microscopy or flow cytometry to evaluate the cellular uptake efficiency of the conjugates [1] - Drug Release Assay: Target cells were treated with Fmoc-PEA-based conjugates, and intracellular components were extracted after a specific incubation time [1] Free active drug was separated and analyzed by chromatographic methods to verify the hydrolysis of the phosphate ester bond of Fmoc-PEA and the subsequent release of the active drug inside cells [1] - Cytotoxicity Assay: Cytotoxic drugs conjugated with Fmoc-PEA were co-cultured with target cells [1] After incubation for a predetermined period, cell viability assays were performed to evaluate the inhibitory activity of the conjugates against target cells, verifying the effectiveness of the Fmoc-PEA-mediated intracellular delivery system [1]
References	[1]. PHOSPHATE BASED LINKERS FOR INTRACELLULAR DELIVERY OF DRUG CONJUGATES. WO2015153401A1.
Additional Infomation	- Fmoc-PEA is a phosphate-based linker with a chemical structure containing an Fmoc (9-fluorenylmethoxycarbonyl) protecting group, a phosphate ester moiety, and a PEA (polyetheramine) segment [1] - Its core application is as a connecting unit in drug conjugates, linking carrier molecules with active drug molecules [1] - The design of Fmoc-PEA ensures stability of the drug conjugate in the bloodstream (no hydrolysis) while enabling efficient intracellular release of the active drug via phosphatase-catalyzed hydrolysis of the phosphate ester bond [1] - It is suitable for intracellular delivery of various cytotoxic drugs and biological agents [1] - The Fmoc group of Fmoc-PEA can be deprotected under specific conditions, facilitating the protection and deprotection of amino groups during the synthesis of drug conjugates [1]

Solubility Data

Solubility (In Vitro)

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

Solubility (In Vivo)

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. Injection Formulations (e.g. IP/IV/IM/SC) 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). 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) Oral Formulations 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). 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 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.7525 mL	13.7627 mL	27.5255 mL
	5 mM	0.5505 mL	2.7525 mL	5.5051 mL
	10 mM	0.2753 mL	1.3763 mL	2.7525 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.