Naphthylfluorescein inhibits the response between HIF-1 and Mint3. Naphthylfluorescein inhibits HIF-1 activity as well as repair and glycolytic activities of macrophages without causing external cytotoxic effect or internal reactions. Naphthylfluorescein is also a fluorophore pH-sensitive probe that can be used for functional Cherenkov imaging.

Physicochemical Properties

Molecular Formula	C28H16O5
Molecular Weight	432.424
Exact Mass	432.099
CAS #	61419-02-1
PubChem CID	3124834
Appearance	Purple to purplish red solid powder
Density	1.6±0.1 g/cm3
Boiling Point	755.5±60.0 °C at 760 mmHg
Melting Point	189ºC (dec.)(lit.)
Flash Point	266.4±26.4 °C
Vapour Pressure	0.0±2.6 mmHg at 25°C
Index of Refraction	1.861
LogP	5.09
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	0
Heavy Atom Count	33
Complexity	740
Defined Atom Stereocenter Count	0
InChi Key	IXQIUDNVFVTQLJ-UHFFFAOYSA-N
InChi Code	InChI=1S/C28H16O5/c29-17-7-9-19-15(13-17)5-11-23-25(19)32-26-20-10-8-18(30)14-16(20)6-12-24(26)28(23)22-4-2-1-3-21(22)27(31)33-28/h1-14,29-30H
Chemical Name	7',19'-dihydroxyspiro[2-benzofuran-3,13'-2-oxapentacyclo[12.8.0.03,12.04,9.017,22]docosa-1(14),3(12),4(9),5,7,10,15,17(22),18,20-decaene]-1-one
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
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	Naphthylfluorescein (compound 19) inhibits HIF-1 reporter gene activity in a concentration-dependent manner (0-10 μM; 24 hours) [1].
ln Vivo	Naphthafluorescein (Compound 19) (100 mg/kg; i.p.; once daily for 5 days followed by 2 days off for 2 weeks) does not induce weight loss or major histological abnormalities in the lungs, liver, and kidneys, or Causes severe symptoms and harmful consequences in mice for at least two weeks [1]. Naphthofluorescein (compound 19) (100 mg/kg; i.p.; once daily for 5 days, followed by 2 days of rest for 2 weeks) greatly decreased tumor growth of subcutaneously injected E0771 cells and considerably diminished MDA- Tumor growth of MB-231 and MDA-MB-231. AsPC-1 cells in immunodeficient mice. In turn, naphthylfluorescein did not decrease tumor growth in FIH-1-depleted MDA-MB-231 cells [1]. Naphthafluorescein (compound 19) (100 mg/kg; i.p.; once daily for 5 days followed by 2 days rest for 2 weeks) suppresses tumor development in human cancer cells in a FIH-1-dependent way [1] .
Cell Assay	Cell viability assay [1] Cell Types: HT1080 Cell Tested Concentrations: 0-10 μM Incubation Duration: 24 hrs (hours) Experimental Results: Dramatically inhibited HIF-1 reporter gene activity in a concentration-dependent manner.
Animal Protocol	Animal/Disease Models: Male C57BL/6J mice (8 weeks) [1] Doses: 100 mg/kg Route of Administration: intraperitoneal (ip) injection; one time/day for 5 days, then rest for 2 days for 2 weeks. Experimental Results: Mice were There was no weight loss or obvious histological abnormalities in the lungs, liver, or kidneys, or serious adverse reactions for at least 2 weeks. Animal/Disease Models: Female C57BL/6J mice (E0771) and BALB/c nude mice (HT1080, MDA-MB-231 and AsPC-1) (6 weeks) [1] Doses: 100 mg/kg Route of Administration: intraperitoneal (ip) injection ; one time/day for 5 days, then 2 days off for 2 weeks. Experimental Results: Dramatically inhibited tumor growth of subcutaneously (sc) (sc) injected E0771 cells and Dramatically attenuated the growth of MDA-MB-231 and AsPC-1 cells in immunodeficient mice. Tumor growth. In turn, naphthylfluorescein did not attenuate tumor growth in FIH-1-depleted MDA-MB-231 cells.
References	[1]. Pharmacological inhibition of Mint3 attenuates tumour growth, metastasis, and endotoxic shock. Commun Biol. 2021;4(1):1165. Published 2021 Oct 7. [2]. Development of fluorinated naphthofluoresceins for Cerenkov imaging. J Fluor Chem. 2019;225:27-34.
Additional Infomation	Naphthofluorescein is a member of xanthenes.

Solubility Data

Solubility (In Vitro)

DMSO : ~125 mg/mL (~289.07 mM)

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	2.3126 mL	11.5628 mL	23.1257 mL
	5 mM	0.4625 mL	2.3126 mL	4.6251 mL
	10 mM	0.2313 mL	1.1563 mL	2.3126 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.