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7 protocols using anhydrous ch2cl2

1

Characterization of Organic Compounds

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UV–visible–NIR
spectra were recorded on an HP 8453 spectrophotometer. 1H NMR spectra were recorded on 400 MHz Bruker Avance III HD spectrometer
equipped with a 5 mm BB/1H SmartProbe at 253 K in CDCl3 or CD2Cl2 and referenced to residual
CHCl3 at 7.26 ppm and CH2Cl2 at 5.31
ppm. Mass spectra were recorded on a Thermo Scientific LTQ Orbitrap
XL spectrometer with an ION-MAX electrospray ion source in positive
mode.
Cyclic voltammetry was carried out at 298 K with an EG&G
Model 263A potentiostat having a three-electrode system–a glassy
carbon working electrode, a platinum wire counterelectrode, and a
saturated calomel reference electrode (SCE). Anhydrous CH2Cl2 (Aldrich) was used as solvent and tetrakis(n-butyl)ammonium perchlorate, recrystallized twice from
absolute ethanol, and dried in a desiccator for at least 2 weeks,
was used as the supporting electrolyte. The reference electrode was
separated from the bulk solution using a fritted-glass bridge filled
with the solvent/supporting-electrolyte mixture. The electrolyte solution
was purged with argon for at least 2 min, and all measurements were
carried out under an argon blanket. All potentials were referenced
to the SCE. Elemental analyses were obtained from Atlantic Microlab,
Inc.
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2

Spectroscopic and Electrochemical Analysis

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UV–visible
spectra
were recorded on an HP 8454 spectrophotometer. 1H NMR spectra
were recorded on a 400 MHz Bruker AVANCE III HD spectrometer equipped
with a 5 mm BB/1H SmartProbe and referenced to either residual CH2Cl2 at 5.32 ppm or residual CHCl3 at
7.26 ppm. High-resolution electrospray-ionization mass spectra were
recorded on an Orbitrap Exploris 120 spectrometer using methanolic
solutions.
Cyclic voltammetry was carried out at ambient temperature
with a Gamry Reference 620 potentiostat equipped with a three-electrode
system: a glassy carbon working electrode, a platinum wire counter
electrode, and a saturated calomel reference electrode (SCE). Tetra(n-butyl)ammonium perchlorate (CAUTION!)
was used as the supporting electrolyte. Anhydrous CH2Cl2 (Aldrich) was used as the solvent. The electrolyte solution
was purged with argon for at least 2 min prior to all measurements,
which was carried out under an argon blanket. All potentials were
referenced to the SCE.
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3

UV-vis, NMR, and Electrochemical Analyses

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UV–vis
spectra were recorded
on a Cary 8454 spectrophotometer. 1H NMR spectra were recorded
on a 400 MHz Bruker Avance III HD spectrometer (equipped with a 5-mm
BB/1H SmartProbe) at a temperature of 298 K in tetrahydrofuran
(THF)-d8 and referenced to residual THF
protons at 3.48 and 2.37 ppm. High-resolution (HR) mass spectra were
recorded (typically in the positive-ion mode) on a Thermo LTQ Orbitrap
XL spectrometer equipped with an electrospray ION MAX source. Elemental
analyses were performed by Atlantic Microlab, Inc.
Cyclic voltammetry
was carried out at 298 K with an EG&G model 263A potentiostat
having a three-electrode system: a glassy carbon working electrode,
a platinum wire counterelectrode, and a standard calomel reference
electrode (SCE). Tetra-n-butylammonium perchlorate,
recrystallized twice from absolute ethanol and dried in a desiccator
for at least two weeks, was used as the supporting electrolyte. Anhydrous
CH2Cl2 (Sigma-Aldrich) was used as a solvent.
The reference electrode was separated from the bulk solution by a
fritted-glass bridge filled with a saturated AgCl/KCl mixture. The
electrolyte solution was purged with argon for at least 2 min, and
all measurements were carried out under an argon blanket. All potentials
were referenced to the SCE.
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4

Synthesis and Characterization of COP

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2-Chloro-2-oxo-1,3,2-dioxaphospholane (COP) was synthesized, distilled under vacuum and stored at −20 °C.23 (link) Tetrahydrofuran (THF) purchased from Alfa Aesar in Shanghai was used after distillation. After purification by phthalic anhydride, sodium hydroxide and calcium hydride, triethylamine (TEA, Aladdin) was distilled for subsequent studies. Monomethoxy poly(ethylene glycol) (mPEG, Mn = 5000 g mol−1) was acquired from Sigma-Aldrich and was dried by azeotropic distillation with toluene subsequently. 1,5,7-Triazabicyclo[4.4.0]dec-5-ene (TBD), anhydrous CH2Cl2, 3-butyn-1-ol, DOX and Ce6 were purchased from Sigma-Aldrich. All other undeclared reagents and solvents were of analytical grade and used as received.
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5

Synthesis of Photosensitizer-Conjugated Nanocarriers

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mPEG-OH, anhydrous CH2Cl2, docosanedioic acid 6b, KI, dexamethasone and streptomycin were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Triethyl amine and anhydrous DMSO were purchased from Acros Organics. Mesyl chloride, I2, 11-mercaptoundecanoic acid 5a and N-hydroxysuccinimide (NHS) were purchased from Aldrich. NH3 25% solution, NaSO4, MeOH, Na2SO3 and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDCI) were purchased from Carl Roth (Carl Roth GmbH + Co. KG, Karlsruhe, Germany). NaOH and HCl were bought from Fisher Scientific (Fisher Scientific GmbH, Schwerte, Germany). 2-Morpholinoethanesulfonic acid (MES), GSH, oxidized glutathione (GSSG), TCEP and fluorescein isothiocyanate (FITC) were purchased from TCI (TCI Deutschland GmbH, Eschborn, Germany). Hydroxyethyl cellulose (HEC) gel was purchased from Ashland (Düsseldorf, Germany). Diethyl ether, acetone and acetonitrile were purchased from VWR Chemicals (Avantor, Darmstadt, Germany). Meso-tetra (m-hydroxyphenyl) porphyrin (mTHPP) was purchased from Biolitec (Jena, Germany). Pheophorbide A (PhA) was purchased from Frontier scientific (Logan, UT, USA). Rapamycin was bought from LC Laboratories (Woburn, MA, USA). Aminated hyperbranched polyglycerol (hPG-NH2, 10 kDa, 70% functionalized, 20 mg/mL in MeOH) was synthesized according to Roller et al. [18 (link)].
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6

Purification of Organic Compounds

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Anhydrous CH2Cl2, THF, CH3CN, DMF, DMA, acetone and other solvents were reagent grade and purchased from Sigma-Aldrich (Milwaukee, WI) or Fisher (Pittsburgh, PA). Analytical TLC was performed using UNIPLATE silica gel GHLF 250 μm pre-coated plates (ANALTECH, Newark, DE). Column chromatography was performed using silica gel (200–400 mesh, 60 Å) from Sigma-Aldrich. Chemical reactions sensitive to air or moisture were carried out under nitrogen atmosphere in oven-dried glassware. Reagent solutions, unless otherwise noted, were handled under a nitrogen atmosphere using syringe techniques. Flash chromatography was performed using Teledyne ISCO (Lincoln, NE) Combiflash RF system and disposable normal silica cartridges of 30–50 μ particle size, 230–400 mesh size and 60 Å pore size. The flow rate of the mobile phase was in the range of 18 to 35 ml/min and mobile phase gradients of ethyl acetate/hexanes and CH2Cl2/CH3OH were used to elute compounds.
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7

Electrochemical Characterization of Monomer

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Cyclic voltammetry
(CV) and differential
pulse voltammetry (DPV) experiments were performed in a one-compartment
three-electrode electrochemical cell with a glassy carbon disk (diameter
= 2 mm) working electrode, platinum counter electrode, and an Ag/0.1
M AgNO3/CH3CN reference electrode, whose potential
was verified at the end of each set of experiments using the ferrocene
couple (Fc/Fc+). The monomer concentration was 1 ×
10–3 M in an electrolyte consisting of 0.1 M Bu4NBF4 (for electrochemical analysis, Sigma-Aldrich)
dissolved in anhydrous CH2Cl2 (Sigma–Aldrich).
For all electrochemical experiments, Autolab PGSTAT 20 (Eco Chemie,
Netherlands) was used.
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