Bright shining single-chain nanoparticles: Advanced applications in photocatalysis and photodynamic therapy

B igh Shining Single-Chain
Nanopa icles: Ad anced
Applica ions in Pho oca alysis and
Pho odynamic The apy
by Da ide A ena
Supe ised by P o . José A. Pomposo and D . M. Es e Ve de
Donos ia - San Sebas ián, 2024
(cc) 2024 Da ide A ena (cc by-nc-nd 4.0)
Acknowledgemen s
Fi s ly, I since ely acknowledge my hesis di ec o s P o . José A. Pomposo and
D . M. Es e Ve de. Wi h wise guidance and daily suppo , you bo h made me g ow as
a esea che and as a pe son.
I g a e ully acknowledge he Ma e ial Physics Cen e and he Polyme s, So
Ma e & Sus ainable Ma e ials G oup o gi ing me he oppo uni y o ca y ou my
doc o al s udies in a scien i ically s imula ing en i onmen . I would like o especially
acknowledge P o . A an xa A be and D . Amaia I u ospe o he con inue dedica ion
and suppo wi h he SAXS cha ac e iza ion.
I would like exp ess deep g a i ude o D . Magali Ga y-Bobo, no only o
gi ing me he oppo uni y o wo k on an essen ial pa o his wo k a he Uni e si é de
Mon pellie , bu also o he kindes welcoming and con inuous suppo . I would also
like o ex end my mos since e g a i ude o D . Ch is ophe Nguyen, you eachings a e,
in a wo d, i eplaceable. Thank you o all he Glyco and Nano ec o s o The apeu ic
Ta ge ing G oup, Lamiaa, Lau e, Ma iana, Nadi , Alain, Denis, Kamel, Ma ie, Khaled,
Melanie, o ans o ming my s ay in such a meaning ul and en iching expe ience.
I would also like o g a e ully acknowledge P o . Fabienne Dumoulin, P o .
Zo aida F eixa, D . Ane Izaskun A anbu u Lei a and D . I án Ri illa, o hei
c ucially impo an con ibu ions and suppo o he wo k which is now condensed in
his hesis.
I would like o hank my amily and Daniel, o unde s anding and always
being he e, belie ing in me, b inging inex inguishable ligh o my li e. This p ojec
would ha e no been possible wi hou you.
Finally, hank o my b o he , his hesis is dedica ed o you, o he ime ha ,
wi h my absence, I s ole you.
1
Summa y
Keywo ds: Single-Chain Nanopa icles, Pho ochemis y,
Pho oca alysis, Pho odynamic The apy, Func ional Polyme s.
Wi h he wo k which we p esen in his hesis, we aimed o es ablish a link
be ween single-chain nanopa icle (SCNP) echnology and pho oca alysis, inding
no el and ad anced applica ions o nex -gene a ion, ligh -ha es ing SCNPs. In
pa icula , we ocused on employing he oppo uni ies o e ed by SCNP unique
opology o enable ad anced applica ions o pho oca alysis in aqueous and complex
en i onmen s such as o o ganic pho oca alysis and pho odynamic he apy (PDT) o
cance . The p esen hesis’ s uc u e comp ises wo i s Chap e s, in which basic
concep s and ecen li e a u e on bo h SCNPs echnology and o ganic pho oca alysis
a e b ie ly e iewed, opening he discussion o he expe imen al esul s disclosed in
Chap e III and IV.
Speci ically, in Chap e I he de ini ion o SCNPs is gi en and he ele an
syn he ic aspec s o hei p epa a ion a e illus a ed. Namely, he syn hesis o he
polyme ic p ecu so s ia e e sible-addi ion agmen a ion eac ion (RAFT)
polyme iza ions and he main s a egies o chain olding / collapse epo ed in
li e a u e a e discussed. In conclusion o he i s Chap e we delinea ed he main aims
o he p esen wo k, and inally ou lined some majo con ibu ions o he ields o
ca alysis and nanomedicine in ol ing SCNPs-based sys ems.

2
In Chap e II, we in oduce he de ini ions, he undamen al concep s o
pho oca alysis, wi h a special ocus on o ganic pho oca alyzed eac ions and he majo
cons ain s p esen ed by he use o wa e as sol en o ca y ou such kind o aluable
ans o ma ions. Apa om i s , seminal epo s a he in ancy o O ganic Chemis y,
in which wa e was commonly used as eac ion medium, he aqueous en i onmen
quickly disappea ed amongs he common p ac ice o syn he ic o ganic chemis s o e
he cou se o he pas cen u y. Conside ing he g owing demand o inc easingly low-
impac p ocesses, bo h om an economic and ecological poin o iew, he use o wa e
o eplace lammable, o en oxic and expensi e o ganic sol en s in indus ially
ele an p ocesses, e.g. he aluable small o ganic molecules p epa a ion, ecen ly
gained back in e es amongs he scien i ic communi y. Fo his eason, in Chap e II
we pu special a en ion on how sup amolecula app oaches ha e been ecen ly applied
o add ess he issues associa ed wi h he use o wa e as sol en o pho o-induced
o ganic ans o ma ions.
In Chap e III, we con inue he discussion epo ing he p epa a ion o a no el
class o e sa ile SCNP capable o e icien ly ca y ou pho oca aly ic o ganic eac ions
in wa e . We designed an amphiphilic polyme ic p ecu so o de ined molecula weigh
and dispe si y exploi ing he amphiphilic polyme ic sca old o he
Poly[(olygoe hylene glycol)monome hyle he me hac yla e]- -Poly(ace oace oxye hyl
me hac yla e), Poly(OEGMA)- -Poly(AEMA), which was p epa ed by RAFT
copolyme iza ion o he comme cially a ailable monome s OEGMA and AEMA. The
ob ained copolyme ic p ecu so was hen unc ionalized and gi en o pho oca aly ic
ac i i y by deco a ion wi h an i idium(III)-based cyclome ala ed complex hough a
mild pos -polyme iza ion unc ionaliza ion app oach, exploi ing he ich β-ke oes e
chemis y o he hyd ophobic comonome AEMA. The p epa ed pho oac i e
amphiphile copolyme esul ed o e icien ly sel -assemble in aqueous solu ion by
olding / collapse in o a SCNP s uc u e as e ealed by dynamic ligh sca e ing (DLS)
echniques and as con i med by UV-Visible spec opho ome y. In e es ingly, he
wa e soluble, i idium(III)-con aining SCNPs, which we called a i icial pho osyn hase
(APS) allowed he obse a ion o an enhancemen in he pho oluminescence (PL) in
wa e wi h espec o he o ganic sol en solu ions, sugges ing he a ising o
3
agg ega ion-induced emission phenomena a ising om he locally compac
hyd ophobic pocke s o he SCNP.
Wi h he APS in hand, we subsequen ly es ed hei abili y o pe o m a a ie y
o o ganic eac ions. Speci ically, we we e able o obse e e icien isible-ligh
induced pho oca aly ic ac i i y o wo unp eceden ly epo ed o ganic eac ions in
wa e , namely he pho o[2+2]cycloaddi ion o inyl a enes and he α-a yla ion o
a ylamines, as well as he oxida ion o 9-subs i u ed an h acenes and he β-
sul onyla ion o s y ene-like compounds. Due o he simila i ies o hese APS o
enzymes, kine ics da a o he pho o[2+2]cycloaddi ion o inyl a enes “in wa e ”
pho oca alyzed by APS we e analyzed in e ms o he adi ional Michaelis-Men en
model. The appa en alues o kca and KM ob ained we e 2.6 s-1 and 4.6 × 10-2 M,
espec i ely, alues which a e compa a i ely and signi ican ly close o wha epo ed
o some bio ic enzymes (Chymo ypsin shows kca = 0.14 s-1 and KM = 1.5 × 10-2 M,
Pepsin kca = 0.50 s-1 and KM = 3.0 × 10-4 M, and RNA syn he ase kca = 7.6 s-1 and KM
= 9.0 × 10-4 M).
In summa y, in Chap e III we desc ibe and epo a i s gene a ion o APS,
b oadening he possibili ies o pe o ming challenging “in wa e ” o ganic
ans o ma ions ia APS-media ed isible-ligh pho oca alysis.
In Chap e IV, we epo he design and syn hesis o a polyme ic p ecu so s o
enhance he PDT e iciency o a no el, long-wa eleng h abso bing zinc(II)-
ph halocyanine (ZnPc). Fo his, we ook ad an age o he well-known sel -assembly
capabili y o an h acene molecules, we p epa ed an h acene-based amphiphilic
copolyme s om he comme cially a ailable hyd ophobic monome 9-
an h acenylme hyl me hac yla e (AnMA) and he hyd ophilic OEGMA ia RAFT
copolyme iza ion. The p epa ed Poly(AnMA)-co-Poly(OEGMA)s esul ed o be bo h
capable o sel -assembly in wa e and o e icien ly encapsula ing he a - ed-
esponsi e complex ZnPc, yielding s able, wa e soluble, ed-ligh eac i e SCNPs,
which we called a i icial pho o-oxidases (APO), mainly due o hei abili y o induce
oxida i e s ess in cell upon exposu e o ligh and o hei ul a-small dimensions (<
20 nm). The nano-objec s we e cha ac e ized bo h by DLS and SAXS, which e ealed
4
he e ec o he ZnPc encapsula ion on he s e ic hind ance o he SCNPs co e h ough
he measu emen o he adius o gy a ion in p esence and / o absence o ZnPc.
In e es ingly, he eadily p epa ed nano-assemblies showed di e en pho oluminescen
p ope ies in he ed egion depending on he o e all an h acene mola ac ion in he
polyme ic p ecu so . In pa icula , he ex en o ei he b oadening o quenching o Q-
band ansi ions o ZnPc inc eased upon dec easing he an h acene mola ac ion in
he nanoca ie . Analogously, a p onounced quenching (λexc = 650 nm) o he ZnPc
emission in he a ed is obse ed upon dec easing he an h acene con en , hough
allowing he unabili y o he deg ee o agg ega ion wi hin he hyd ophobic co e o he
nanoca ie .
We inally es ed APO-ZnPc agains human b eas cance cell MDA-MB-231
lines o assess hei PDT e iciency. Ha ing obse ed ou s anding pe o mance o one
o he selec ed o mula ions, we inally p o ed hei PDT ac i i y in zeb a ish emb yo
xenog a s as a mo e accu a e human cance model.
In conclusion, in he p esen hesis, he de elopmen o no el sys ems based on
SCNPs o ad anced applica ions in pho oca alyzed o ganic eac ions and
pho odynamic he apy has been s udied and ca ied ou , demons a ing ha he
echnology o single-polyme ic chains olding can be exploi ed o he ab ica ion o
a i icial nano-objec s wi h p o ein- esembling s uc u e o ailo ed pho oca aly ic
ac i i y.
5
Resumen
Palab as cla e: Single-Chain Nanopa icles, Fo oquímica,
Fo oca álisis, Fo o e apia Dinámica, Políme os Funcionales.
Median e el p esen e abajo, se p e ende es ablece una conexión en e la
ecnología de nanopa ículas polimé icas unimolecula es (SCNP, del inglés single-
chain nanopa icles) y la o oca álisis, encon ando aplicaciones no edosas y
a anzadas pa a las SCNP de nue a gene ación capaces de u iliza la luz como uen e
de ene gía. En conc e o, se ha a ado de ap o echa las ca ac e ís icas opológicas de
las SCNP pa a implemen a aplicaciones a anzadas de o oca álisis en ambien es
acuosos y complejos, con el obje i o de emplea las en casos como la o oca álisis
o gánica y la e apia o odinámica (PDT, del inglés pho odynamic he apy) del cánce .
La es uc u a de la p esen e esis cons a de dos p ime os capí ulos, en los que se e isan
b e emen e los concep os básicos y la bibliog a ía ecien e an o de la ecnología de las
SCNPs como de la o oca álisis o gánica, ab iendo la discusión pa a los esul ados
expe imen ales expues os en los capí ulos III y IV.
Conc e amen e, en el Capí ulo I se da la de inición de SCNPs y se ilus an los
p incipales aspec os sin é icos pa a su p epa ación. En conc e o, se discu e la sín esis
de los p ecu so es polimé icos median e polime izaciones po adición, agmen ación
y ans e encia e e sible (RAFT, del inglés e e sible addi ion- agmen a ion
ans e ) y las p incipales es a egias pa a el plegamien o / colapso de cadenas desc i as
12
2.2.4. So Polyme ic Ma e ials o Aqueous O ganic Pho oca alysis
77
2.3. Conclusions
79
2.4. Re e ences
81
3. Chap e III
89
3.1. In oduc ion
89
3.2. Main Aims
90
3.3. Resul s and Discussion
91
3.3.1. P epa a ion o he pho oac i e polyme s
91
3.3.2. Sel -assembly in aqueous solu ion
94
3.3.3. Pho oca aly ic ac i i y “in wa e ”
96
3.3.4. E ec o APS ype on con e sion
103
3.3.5. Kine ic aspec s
104
3.3.6. Recyclabili y o APS
108
3.4. Expe imen al Techniques
110
3.4.1. Sol en s and eagen s
110
3.4.2. Analy ical me hods and echniques
111
3.4.3. Syn hesis and cha ac e iza ion o compounds
114
3.4.3.1. Syn hesis o he polyme ic p ecu so Poly(OEGMA300-co-
AEMA) (P1)
114
3.4.3.2. Syn hesis o he Hyd oxo-B idged I idium(III) Dime
Te akis(2-phenylpy idina o-N,C2)(m-dihyd oxy)dii idium(III)
([I (ppy)2OH]2) (C1)
115
3.4.3.3. Syn hesis o he Cyclome ala ed Complex Bis[2-(2-py idinyl-
N)phenyl-C](me hyl ace oace a o)i idium(III) (C2)
116
3.4.3.4. Syn hesis o I idium(III)-Deco a ed Copolyme s a Di e en
I idium(III) Loadings (P1-I 40, P1-I 23 and P1-I 10)
117
3.4.3.5. P epa a ion o A i icial Pho o-Syn hases based on I idium(III)-
Deco a ed Single Chain Nanopa icles in Aqueous Solu ions (APS-I 40, APS-
I 23, APS-I 10)
118
3.4.4. “In Wa e ” [2+2] Pho ocycloaddi ion o Vinyl A enes
119

13
3.4.5. “In Wa e ” Oxida ion o 9-Subs i u ed An h acenes
122
3.4.6. “In Wa e ” α-A yla ion o A ylamines
123
3.4.7. “In wa e ” β-Hyd oxysul onyla ion o α-Me hyl S y ene
126
3.5. Conclusions
128
3.6. Re e ences
129
4. Chap e IV
137
4.1. In oduc ion
137
4.1.1. Pho odynamic The apy
138
4.1.2. T adi ional Pho osensi ize s o PDT
141
4.1.3. Nanos uc u es in PDT
143
4.2. Objec i es
148
5.3. Resul s and Discussion
152
4.3.1. P epa a ion o π-π sel -assembled amphiphilic SCNPs con aining
Zn(II)-ph halocyanine ZnPc
152
4.3.1.1. P epa a ion o he nanoca ie s
152
4.3.1.2. P epa a ion o he pho osensi ize ZnPc
156
4.3.1.3. Encapsula ion o he pho osensi ize ZnPc
158
4.3.2. In i o Imaging and PDT wi h Amphiphilic SCNPs Con aining
ZnPc Molecules and An h acene Moie ies
161
4.3.3. π-π sel -assembled amphiphilic SCNPs con aining Zn(II)-
ph halocyanine Pc as imaging and a - ed pho o-killing agen s o PDT in
Zeb a ish emb yo xenog a s
168
4.4. Expe imen al Techniques
171
4.4.1. Ma e ials
171
4.4.2. Syn hesis o he pho osensi ize ZnPc
171
4.4.3. Syn hesis o P1
171
4.4.4. Syn hesis o P2
172
4.4.5. Syn hesis o P3
172
4.4.6. P epa a ion o APOx-Pcy
173
4.4.7. Cell cul u e condi ions o in i o expe imen s
173
14
4.4.8. Cell iabili y assay
174
4.4.9. In i o da k cy o oxici y
174
4.4.10. In i o pho o oxici y assay
174
4.4.11. Reac i e oxygen species (ROS) p oduc ion
175
4.4.12. Danio Re io emb yos handling o in Zeb a ish expe imen s
175
4.4.13. Injec ion, i adia ion, and imaging o MDA-MB-231 in Zeb a ish
emb yos
176
4.5. Conclusions
177
4.6. Re e ences
178
5. Conclusions
187
Appendix o Chap e III
193
A.III.1. SEC ch oma og ams
193
A.III.2. DLS da a
196
A.III.3. Supplemen a y UV-Vis spec a
202
A.III.4 NMR spec a
203
Appendix o Chap e IV
236
A.IV.1. p epa a ion o copolyme s wi h highe an h acene con en
236
A.IV.1.1. Syn hesis o he copolyme P4
236
A.IV.1.2. Syn hesis o he copolyme P5
236
A.IV.2. Room-ligh PDT expe imen
237
A.IV.3. SEC ch oma og ams
238
A.IV.4. NMR spec a
241
A.IV.5. Supplemen a y spec oscopic da a
245
15
Lis o Abb e ia ions
[I (ppy)2OH]2
Dihyd oxo e akis[2-(2-py idinyl)phenyl]dii idium-(III)
dime
°C
Celsius deg ee / deg ees
µL
Mic oli e / mic oli e s
µmol
Mic omole / mic omoles
1Sn
Single s exci ed s a es
2-CNIPB
2-chlo o-N,N-diisop opylbenzamide
Å
Angs om / angs oms
AA
α-A yla ion o a ylamines
ABA
4-ace oxybenzaldehyde
AcOE
E hyl ace a e
AEE
Agg ega ion enhanced emission
AEMA
4-ace oace oxye hyl me hac yla e
AFM
A om- o ce mic oscopy
AgCl
Sil e chlo ide
AgOT
Sil e (I) i luo ome hansul ona e
AIBN
Azobisisobu y oni ile
APS
A i icial pho osyn hase
ATRP
A om- ans e adical polyme iza ion
AuNP
Gold nanopa icle
BHT
Bu yla ed hyd oxy oluene
BODIPY
4,4-di luo o-4-bo a-3a,4a-diaza-s-indacenes
c%
Con e sion
CA
[2+2] Cycloaddi ion o inyl a enes
cal
Calo y / calo ies
CDCl3
Deu e a ed chlo o o m
CH2Cl2
Me hylene chlo ide
CHCl3
Chlo o o m
cm
Cen ime e / cen ime e s
16
COSY
Homonuclea co ela ion spec oscopy
CPADB
4-cyano-4-( hiobenzoyl hio)pen anoic acid
CRP
Con olled adical polyme iza ions
CTA
Chain- ans e agen
CTAB
Ce immonium b omide
d
in a-pa icle pho oca aly ic uni densi y
Đ
Dispe si y
Da
Dal ons (u.m.a.)
DCB
1,4-dicyanobenzene
DEPT
Dis o ionless enhancemen by pola iza ion ans e
Dh
Hyd odynamic diame e
DLS
Dynamic ligh sca e ing
DMA
Dime hylace amide
DMEM
Dulbecco’s Modi ied Eagle’s Medium
DMF
N,N-dime hyl o mammide
DMSO
Dime hyl sul oxide
DP
Deg ee o polyme iza ion
DPBF
1,3-diphenylisobenzo u an
DRI
Di e en ial e ac i e index
E*ox
Exci ed s a e’s oxida ion po en ial
E* ed
Exci ed s a e’s educ ion po en ial
Eox
Oxida ion po en ial
EPR
Enhanced pe meabili y and e en ion
E ed
Reduc ion po en ial
ET
Ene gy ans e
E 2O
Die hyl e he
E OH
E hanol
FRET
Fö s e esonance ene gy ans e
FT-IR
Fou ie ans o m in a ed
GPC
Gel pe mea ion ch oma og aphy
h
Hou / hou s
HPAM
N-(2-hyd oxyp opyl)ac ylamide
HPLC
High p essu e liquid ch oma og aphy
HS
α-S y ene β-hyd oxysul onyla ion
HSQC
He e onuclea single quan um cohe ence spec oscopy
IONP
I on oxide nanopa icle
IR
In a ed
I (ppy)2(acac)
Bis(2-phenylpy idine)-(ace ylace ona e)i idium(III)
I (ppy)2(meacac)
bis[2-(2-py idinyl-N)phenyl-C](me hyl
ace oace a o)i idium(III)
17
I Cl3
I idium(III) ichlo ide
I sppy
ac- is[2-(5’-sul ona ophenyl)py idine]i ida e(III)
ISC
In e -sys em c ossing
IUPAC
In e na ional Union o Pu e and Applied Chemis y
J
Joule / joules
JNPs
Janus nanopa icles
K2CO3
Po assium ca bona e
kca ,app
Appa en ca aly ic cons an
kDa
Kilodal on / kilodal ons
KM,app
Appa en Michaelis-Men en cons an
kV
Kilo ol / kilo ol s
L
Li e / li e s
LAM
Less ac i a ed monome
LED
Ligh -emi ing diodes
LI
i idium loading
M
Mola i y / mola
m
Me e / me e s
MA
Me hyl ac yla e
mA
Milliampe e / milliampe es
MALDI
Mass analysis lase deso p ion ioniza ion
MALS
Mul i-angle lase ligh sca e ing
MAM
Mo e ac i a ed monome
MDO
2-me hylene-1,3-dioxepane
MeOH
Me hanol
mg
Millig am / millig ams
MgSO4
Magnesium sul a e
MHz
Megahe z
min
Minu e / minu es
mL
Millili e / millili e s
mm
Millime e / millime e s
MMA
Me hyl me hac yla e
Mn
Numbe a e age molecula weigh
mol
Mole / moles
MPEG
Poly(e hylene glycol e he ) me hyl e he
MTT
4,5-dime hyl hiazol-2-yl)-2,5-diphenyl e azolium b omide
Mw
Weigh ed a e age molecula weigh
mW
Milliwa / milliwa s
Na2HPO4ꞏnH2O
Sodium phospha e dibasic n-hyd a e
NaCl
Sodium chlo ide
NaOAc
Sodium ace a e

18
NaOH
Sodium hyd oxide
NBD
Ni obenzoxadiole
NHSMA
N-hyd oxysuccinimide es e
nm
Nanome e / nanome e s
NMP
Ni oxide-media ed polyme iza ion
NMR
Nuclea magne ic esonance
NVC
N- inyl ca bazole
NVP
N- inyl py olidone
OA
Oxida ion o 9-subs i u ed an h acenes
OD
Op ical densi y
OEGMA
(Oligoe hylene glycol monome hyl e he ) me hac yla e
PC
Pho oca alys
PC*
Pho oca alys exci ed s a e
PC-SCNP
Pho oca aly ic single chain nanopa icles
PDI
Polydispe sion index
PDMEAMA
Poly(dime hylaminoe hylme hac yla e)
PDT
Pho odynamic he apy
PEG
Polye hylene glycol
PL
Pho oluminescence
PMAA
Poly(me hac ylic acid)
PN
Phenoxazine
PNIPAM
Poly(N-isop opylac ylamide)
ppm
Pa s pe million
PS
Pho osensi ize
PVBC
Poly( inylbenzylchlo ide)
PVP
Poly inylpi olidone
. .
Room empe a u e
RAFT
Re e sible-addi ion- agmen a ion chain- ans e
RDRP
Re e sible-deac i a ion adical polyme iza ion
Rg
Radius o gy a ion
Rg
Radius o gy a ion
Rh
Hyd odynamic adius
ROS
Reac i e-oxygen species
ROP
Radical ing opening polyme iza ion
SAPC
Sup amolecula assembly pho oca alys
SAXS
Small-angle X- ay sca e ing
SCE
Sa u a ed calomel elec ode
SCNP
Single-chain nanopa icles
SDS
Sodium dodecyl sul a e
SEC
Size exclusion ch oma og aphy
19
SET
Single-elec on ans e
SLA
S a -like agg ega es
S y
S y ene
TEA
T ie hylamine
THF
Te ahyd o u an
TLC
Thin laye ch oma og aphy
TMS
Te ame hylsilane
TOF
Time o ligh
UPy
2-u eidopy imidinone
UV
Ul a iole
V
Vol / ol s
VAc
Vinyl ace a e
WA
Wen eib amide
Zn(OAc)2
Zinc diace a e
λ exc max
Maximum exci a ion wa elengh
λmax
Maximum wa eleng h
ν
Size scaling exponen
20
21
1. Chap e I
1.1. In oduc ion
Despi e he nanome e (nm) being an ex emely small uni o leng h – housand
million (10-9) o a me e – he manipula ion o ma e wi h dimensions sized om 1 o
100 nanome e s is a deeply in es iga ed b anch o he echnology we now dispose o ,
namely, he nano echnology. Co e ing om medicine, molecula biology, ene gy
s o age, chemis y, semiconduc o physics, jus o ci e a ew, nano echnology includes
all kinds o scien i ic esea ch, ha ing possibly gained place a he cen e o he
a en ion o he scien i ic communi y wo ldwide, in he nano echnology e a i has
al eady been demons a ed how powe ully ou daily li e can be acili a ed by i s
applica ion. The ema kably ascina ing pic u es p esen ed in Figu e 1.1 gi es us a bi e
o how ex ensi ely he nano echnology is in using ou scien i ic cul u e and li e.
28
Scheme 1.2. Equilib ia be ween adicals and do man species in he con olled
adical polyme iza ions.
A he p esen day, se e al CRP echniques a e a ailable, each elying on
di e en chemis ies, mos amously A om T ans e Radical Polyme iza ion (ATRP),5
Ni oxide-Media ed Polyme iza ion (NMP)6 and Re e sible-Addi ion F agmen a ion
Chain-T ans e Polyme iza ion.7 While o ex eme impo ance o syn he ic
applica ions, o exhaus i ely epo all he mechanis ic aspec s o CRPs is a beyond
he scope o he p esen wo k, o his, in he ollowing discussion only he main
ea u es RAFT polyme iza ion will be men ioned, being his la e he mo e ex ensi ely
CRP echnique employed o he wo ks ha cons i u e his hesis, as i will be disclosed
in he nex chap e s, o he ab ica ion o nex -gene a ion, unc ional SCNPs.
As an icipa ed abo e, RAFT polyme iza ion is one o he mos e sa ile and
powe ul ools o he cons uc ion o polyme ic ma e ials o complex a chi ec u e,
de ined molecula weigh s and low dispe si y.8 RAFT is a obus echnique, showing
g ea e sa ili y o a wide ange o monome s and ole ance owa ds many unc ional
g oups. No ably RAFT does no equi e he use o me als, which is e y desi able in
e ms o o e all p ocess economy, pu i ica ion p o ocols and end-use in biology and
elec onic applica ions, allowing o ge close o milde eac ion condi ions which a e
mo e ypical o uncon olled adical polyme iza ions.4,9
RAFT polyme iza ion equi es he p esence o a adical ini ia o and a chain
ans e agen (CTA), commonly a hioca bonyl-based compound (Figu e 1.4).

29
Figu e 1.4. E ec o he subs i uen R and Z o commonly employed CTAs on he
polyme iza ion o comme cially a ailable monome s. Con inuous a ow indica es high o
good compa ibili y, do ed a ow low compa ibili y and no line sca ce o no compa ibili y.4
See ex o monome s comple e names (MMA, HPAM, S y e c.).
As men ioned abo e, he RAFT p ocess o e s a wide ole abili y owa ds
di e en polyme izable monome s and, o ha o be achie ed, CTAs should be chosen
in ela ion o expe imen al condi ions,10 as he na u e o he Z and R g oups o he
RAFT agen (CTA) is undamen al and can be a ied acco dingly o he ype o he
selec ed monome s. In Figu e 1.4, a schema ized selec ion guide o CTA-monome
pai ing is p esen ed.4 The monome species ha can be used in his ype o
polyme iza ion a e con en ionally di ided in o mo e ac i a ed monome s (MAMs) and
less ac i a ed monome s (LAMs). The i s g oup includes monome s wi h inyl
g oups conjuga ed o a oma ic ings (as s y ene, S y) double bonds, ca bonyl o ni ile
g oups such as ac yla es and me hac yla es (e.g. me hyl me hac yla e, MMA; and
me hyl ac yla e, MA) dienes, o me hac ylamides (e.g. N-(2-
hyd oxyp opyl)ac ylamide HPAM), which can be con olled using di hioes e s /
30
di hiobenzoa es o i hioca bona es as CTAs. On he o he hand, commonly employed
LAMs a e inyl ace a es (VAc), inyl chlo ides and 1-alkenes which ha e double
bonds adjacen o a oms o oxygen, ni ogen, halogens o sa u a ed ca bons (e.g. N-
inyl py olidone, NVP; N- inyl ca bazole, NVC).
As illus a ed in Scheme 1.3, in he RAFT eac ion mechanism an ini ia o
gene a es a adical species ha is eadily added o a monome M, hus o ming a Pn
polyme chain.
Scheme 1.3. RAFT polyme iza ion mechanism.
This is ollowed by he addi ion- agmen a ion s age in which he p opaga ing
chains a e added o he CTA, eleasing a new adical species R·, capable o ini ia ing a
new chain ( e-ini ia ion p ocess). Then, in a ypically sho ime wi h espec o he
decomposi ion o he ini ia o , an equilib a ed exchange is es ablished be ween
do man species, which a e illus a ed as S=CS-Pn and con ibu e o he con olled
cha ac e o he RAFT p ocess, and ac i e chains. In an analogy o wha illus a ed in
Scheme 2, he in e media e adical, Pn-SĊ(Z)S-Pm, can spli in bo h di ec ions,
31
p o iding all polyme chains wi h he same chance o g ow. Te mina ion occu s, as
wi h any adical polyme iza ion, h ough coupling and disp opo ion mechanisms. Is
wo h no ing ha he adical species ha a e o med emain cons an h oughou he
cou se o he eac ion. This esul s in a uni o m dis ibu ion o molecula weigh s and
a Ð close o uni y and, o his o occu , he p opaga ion speed should always be
main ained much lowe han ha o he addi ion- agmen a ion s age, ensu ing ha a
single monome ic uni is inco po a ed, on a e age, a e ew ac i a ion cycles, hus
gene a ing chains o simila polyme iza ion deg ee. The small pe cen age o
e mina ion eac ions, which leads o he o ma ion o dead chains, is ela ed o he
numbe o adicals gene a ed in he ini ial s age acco ding o Equa ion 1.1.9
%𝑑𝑒𝑎𝑑 𝑐ℎ𝑎𝑖𝑛 =1
[𝐶𝑇𝐴]
2𝑓[𝐼]0(1 − 𝑒−𝑘𝑡)+ 1
102
Equa ion 1.1
wi h [CTA] equal o he concen a ion o he chain ans e agen in solu ion,
[I]0 equal o he concen a ion o ini ia o p esen in solu ion a he beginning o he
eac ion, k kine ic cons an o he ini ia o decomposi ion a e in he selec ed
expe imen al condi ions and is he decomposi ion a e o he ini ia o .
No ably, a he end o he p ocess, each RAFT (non-dead) chain will ha e a
ca bonyl hioyl hio (Z-C(C=S)S) g oup and an R g oup a he polyme chain
ex emi ies, which is an essen ial ea u e o he ab ica ion o block copolyme s and
o si e-speci ic pos -polyme iza ion modi ica ions.11 Despi e he obus ness o he
RAFT echnique, i should be emphasized ha hioca bonyl moie ies a e subjec o
undesi able eac ions in he p esence o nucleophilic species such amine impu i ies,
which mus be s ic ly absen in he eac ion en i onmen o , simila ly, i is necessa y
o p o ec any amino unc ions i p esen in he monome s used, as pola chemical
eac ions as aminolysis o he Z-C(C=S)S g oup i e e sibly e mina e he chain
polyme iza ion.
The a io be ween he concen a ion o CTA and adical ini ia o [CTA] / [I]0
is he e o e an impo an pa ame e o conside o con olling he inal p ope ies o
32
he syn hesized ma e ial. I is usually g ea e han one, so ha he numbe o molecules
o he RAFT agen in solu ion is always main ained g ea e han he ee adical species.
While he concen a ion o ee adicals in he sys em is ela ed o he a e o
decomposi ion o he ini ia o , he numbe o chains is, indeed, de e mined by he
amoun o CTA.12 An inc ease in he concen a ion o RAFT agen leads o he
o ma ion o polyme s wi h lowe molecula weigh and lowe Đ. Con e sely,
dec easing he [CTA]/[I] a io leads o a highe con e sion a e bu wo se con ol o e
he inal molecula weigh , inc easing he likelihood o i e e sible e mina ion
eac ions. Fo his eason, while op imizing di e en eac ion condi ions o RAFT
polyme iza ions, i is commonly a good p ac ice o main ain a low [CTA] / [I] o
achie e be e con ol o e he inal molecula weigh dis ibu ions o he p oduc . I is
wo h no ing ha , o achie e a ce ain desi ed molecula weigh is possible also by
s opping he polyme iza ion eac ion a de ined eac ion imes. The deg ee o
polyme iza ion DP h, in ac , is gi en by he ollowing equa ion.
𝐷𝑃𝑡ℎ = [𝑀]0− [𝑀]𝑡
[𝐶𝑇𝐴]0+𝑑𝑓([𝐼]0− [𝐼]𝑡)
Equa ion 1.2
whe e [M]0 and [M] a e he monome concen a ions a he beginning o he
eac ion and a ime , espec i ely; [CTA]0 is he ini ial concen a ion o chain ans e
agen , [I]0 is equal o he concen a ion o ini ia o p esen in solu ion a he beginning
o he eac ion, [I] is equal o he concen a ion o ini ia o p esen in solu ion a he
ime , d is he ac ion o chains esul ing om coupling eac ions, and is he
e iciency o he ini ia o .
As a i s app oxima ion, wi h almos uni a y e iciency o he ini ia o and a
low incidence o e mina ion eac ions, he equa ion can be simpli ied o he ollowing.
𝐷𝑃𝑡ℎ = [𝑀]0− [𝑀]𝑡
[𝐶𝑇𝐴]0
Equa ion 1.3
33
This con ol allows he syn hesis o i ually any desi ed polyme a chi ec u e:
mul i-block, s a , g a , s a is ical, al e na ing, and g adien copolyme s, jus o ci e a
ew (Figu e 1.5).9
Figu e 1.5. Examples o di e en a chi ec u es which can be achie ed by RAFT
copolyme iza ion.
1.3.2. S a egies o SCNP Compac ion
To acili a e he in amolecula c osslinking, a wide ange o unc ional g oups
has been explo ed, u ilizing o example pho ochemis y, me al-complexa ion o non-
co alen in e ac ions, as highligh ed by se e al e iew a icles in which as collec ions
o echniques o SCNPs olding ha e been epo ed.13 He ein, h ee main s a egies
a ailable o polyme single-chain compac ion will be b ie ly men ioned, namely (i)
co alen c oss-linking, (ii) me al-induced c oss-linking and (iii) non-co alen c oss-
linking, by e acing some ep esen a i e and ecen li e a u e examples in he ield.
(i) Co alen c oss-linking.
Thanks o he high dissocia ion ene gies o he co alen ange o chemical
bonds, olding a polyme molecule in o a single-chain nanopa icle ia co alen
chemis y is usually a way o ensu e du abili y, mo phology s abili y and compac ness

34
o he inal p oduc . SCNPs which a e ob ained by means o co alen chemis ies a e
mo e s able han hei non-co alen coun e pa s, he e o e acili a ing hei analysis
ia common ch oma og aphy echniques and educing i e e sible agg ega ion
phenomena upon c owding.
S a ing om he he mal dime iza ion o benzocyclobu ene uni s o a single-
polyme ic chain,14 milde and mo e con olled me hodologies o ab ica ion o co alen
SCNPs ha e been in es iga ed, as amide15 and u ea o ma ion,16 as well as hiol-
Michael u ilizing diac yla e moie ies in he la e al polyme ic chain as elec ophiles o
he c oss-linking, which was success ully ca ied ou bo h in o ganic sol en s and in
wa e o di e en unc ional copolyme s.17-19 1,3-dipola cycloaddi ions ha e also
been u ilized o he o ma ion o SCNPs, ei he exploi ing wo complemen a y
unc ional monome s wi hin he same polyme ic p ecu so o wi h he in oduc ion o
a c osslinke .20-23
Se e al s a egies ha e al eady been epo ed in his sense,24-27 amongs which
he pho ochemically induced collapse epo ed by Thannee u e al. is wo h
men ioning.28 In hei wo k on adpole-shaped SCNPs, he au ho s demons a ed he
use o in e molecula pho o-c osslinking o amphiphilic polyac yla es co-
polyme hac yla es block copolyme s, which we e e icien ly con e ed in o adpole-
shaped SCNPs o a ious mo phologies hanks o he pho o[2+2]cycloaddi ion o he
cinnamoyl moie ies p esen in he la e al chains o he p ecu so s. In e es ingly,
di e en mo phologies we e achie ed depending bo h on he sol en used o he pho o
c oss-linking eac ion and he posi ion o he cinnamoyl moie ies in he main chain,
wi hin he hyd ophilic block o in he hyd ophobic sec ion (Figu e 1.6).
35
Figu e 1.6. a) Tadpole-shaped, amphiphilic SCNPs o di e en mo phologies upon
swi ching p ecu so s’ c oss-linkable uni s posi ion, ob ained ia pho o[2+2]cycloaddi ion o
cinnamoyl moie ies.28 b) FRET-SCNPs syn hesisez by Maag e al. ia eac ion wi h he
dib omo bimane c oss-linke .29 c) Reac ion scheme o he eac ion o diazaylide bond
o ma ion o he p epa a ion o he ul a-s able “S audinge ” SCNPs.30
In an analogous wo k, Zhang e al. epo he olding o a linea polyme ia
pho odime iza ion o cyanos ilbene moie ies pendan s, a o ed by sel -o ganiza ion o
he polyme ic p ecu so ia he non-co alen π-π s acking o ces,31 displays how he
concomi an use o in amolecula non-co alen in e ac ions and pho ochemically
induced co alen bond o ma ion can lead o eliable SCNPs o de ined mo phology.
36
The o ma ion o amide and es e bonds o he chain collapse eac ion in he
ab ica ion o SCNPs is ano he eliable s a egy which ha e been ecen ly employed,
as demons a ed by Maag e al., he collapsed, highly packed polyme ic s uc u e
exhibi ing Fö s e esonance ene gy ans e was achie ed by eac ing a linea
polyme ic p ecu so bea ing ni obenzoxadiole (NBD) uni s and ee ca boxylic
moie ies wi h he luo escen eac an dib omobimane (Figu e 5).29
In he wo k published by Jackson e al., s imuli-deg adable SCNPs we e
ob ained h ough olding o a polyme ic p ecu so , which was ob ained ia adical ing-
opening polyme iza ion ( ROP) o 2-me hylene-1,3-dioxepane (MDO) and
me hac ylic acid N-hyd oxysuccinimide es e (NHSMA) and subsequen ly c oss-
linked by condensa ion eac ion wi h a diamino-c oss-linke .17 As an icipa ed,
co alen ly c oss-linked SCNPs a e especially desi able when s abili y, ei he he mal
o chemical, is equi ed o a speci ic applica ion, o his, he p epa a ion o no el
sys ems as he ul a- obus “S audinge ” SCNPs was ecen ly epo ed, o which he
au ho s ook ad an age o a pen a luo ophenyl-bea ing s y enic copolyme , which was
azida ed as showed in Figu e 1.6 and subsequen ly co alen ly c oss-linked by eac ion
wi h a diphosphine c oss-linke , o gi e he s able azaylide bonds, as showed in Figu e
1.6.30
(ii) Me al-media ed olding / collapse.
Me al-o ganic compounds p esen a ich chemis y, which is o en p esen in
na u ally occu ing p o eins and exploi ed by enzymes no only o he ul ima e
ca aly ic pu poses, bu wi h s uc u al unc ions as well, he “zinc inge ” is a
ep esen a i e phenomenon, being a ecu en s uc u al mo i in a plen y o abundan
p o eins o li ing o ganisms.32 Analogously, he o ma ion o me al-o ganic complexes
ha e been employed o he ab ica ion o SCNPs, as in he case o he p epa a ion o
he he e obime allic SCNPs epo ed by Knö el e al.13 The au ho s exploi ed a
bi unc ional e polyme p ecu so , syn hesized by NMP, con aining wo o hogonal
ligand moie ies, phosphines and phosphine oxide which allowed he acile and selec i e
inco po a ion o he wo me als P (II) and Eu(III) espec i ely, as showed in Figu e 1.7.
37
Figu e 1.7. a) O hogonal complexa ion modali ies employed o he e obime allic
me al-induced SCNP collapse.33 b) Sequen ial and o hogonal me al inse ions wi hin he
s uc u e o a me al- olded SCNP.34 c) SCNPs embedded wi h a e ocene-uni , allowing he
subsequen in oduc ion o Pd(II) ions wi hin he nanopa icle.35
In an analogous wo k, he e obime allic Au(I) / Y(III)-SCNPs we e p epa ed
aking ad an age o he o hogonal chemis ies o iphenylphosphines and ca boxyla e
anions.34 In his la e wo k, he in oduc ion o he i s me al, Au(I), hough
complexa ion wi h phosphines in he la e al chains o a unc ionalized polys y ene
(Figu e 1.7) did no in ol e he collapse o he polyme ic chains, which was ins ead
induced only by in oduc ion o he second me al, Y(III), ia complexa ion wi h h ee
ca boxyla e anion pendan s o he same polyme ic chain.
A hi d example o he e obime allic SCNPs is p o ided by he wo k o Rei h
e al., in which a me hod o olding / collapse o he polyme ic chains o SCNPs based
44
The umo a ge ing capabili y and he clea ance s udies o SCNPs ha e been
e alua ed in xenog a mouse models o di e en kinds o human umo s, and
zeb a ish emb yos.60,61 SCNPs ha e also been ecen ly es ed o a ge ing o speci ic
cells, a c ucial challenge in biomedical applica ions o s ep o wa d d ug selec i i y o
a ious diseases ea men s. E.g. K öge e al. glucose-based SCNPs which, a e
ancho ing on nanodiamonds sca olds, showed e icien selec i e mac ophages
imaging capabili y in i o.62 Speci ic ecep o a ge ing was also u ilized by Baij e al.
o cell labeling and imaging o human b eas cance (SK-BR-3) cells.63
Ano he oppo uni y p o ided by SCNPs o nanomedicine applica ions is he
encapsula ion o ac i e compounds, bo h o imaging and he apeu ic pu poses. The
encapsula ion abili ies o SCNPs ha e been in es iga ed o di e en sys ems, bo h
co alen ly and non-co alen ly o med SCNPs. K öge e al. epo ed a SCNP sys em
ha is able o encapsula e bo h hyd ophobic and hyd ophilic small d ug molecules.64
To conclude, SCNPs ha e shown p omise in in i o s udies and in p elimina y
in i o obse a ions. The in es iga ed sys ems usually main ain good cell iabili ies
and he acile access o di e en unc ional g oups allowed he p epa a ion o selec i e,
ul a-small nano-objec s which p esen ed good malignan cell a ge ing beha io .
Despi e he encou aging esul s, sys ema ic in es iga ions o biocompa ibili y,
biodis ibu ion and beha io o he SCNPs in hei biological applica ions a e equi ed
o un a el he ull po en ial o SCNPs o nanomedical pu poses.
1.5. Main Aims and S uc u e o he Thesis
The p incipal objec i e o his wo k consis ed in building a b idge, bo h
concep ual and empi ical, be ween SCNP-based echnology and pho oca alysis. In
pa icula , he p esen hesis ocused on employing he oppo uni ies o e ed by SCNP
opology o enable pho oca alysis in aqueous en i onmen s.
In Chap e II, we aimed a in oducing he de ini ions, undamen al concep s
and majo cons ain s p esen ed by o ganic pho oca alyzed p ocesses in aqueous
solu ions, wi h special a en ion on how sup amolecula app oaches ha e been ecen ly

45
applied o add ess he issues associa ed wi h he use o wa e as sol en o pho o-
induced o ganic ans o ma ions.
These concep s pu he basis o wha we subsequen ly epo ed in Chap e III,
in which we aimed o p epa e a no el class o e sa ile SCNP capable o e icien ly
ca y ou pho oca aly ic o ganic eac ions in wa e . The sol en selec ed by Na u e o
pe o ming biochemical ans o ma ion is, indeed, wa e , a s a egy which o en elies
on he exploi a ion o he hyd ophobic in e ac ions be ween he nano-ca alys s, he
enzymes, and subs a es. Fo his, we designed an amphiphilic polyme ic p ecu so o
ul a-high molecula weigh and low dispe si y, which was embedded wi h
pho oca aly ic ac i i y by deco a ion wi h an i idium(III)-based cyclome ala ed
complex hough a pos -polyme iza ion unc ionaliza ion app oach. The eadily
p epa ed pho oac i e amphiphile, which esul ed o e icien ly sel -assemble in
aqueous solu ion by olding / collapse in o a SCNP s uc u e, was subsequen ly es ed
o i s capabili y o pe o ming wo unp eceden ly epo ed o ganic eac ions in wa e ,
namely he pho o[2+2]cycloaddi ion o inyl a enes and he α-a yla ion o a ylamines,
as well as he oxida ion o 9-subs i u ed an h acenes and he β-sul onyla ion o α-
s y ene.
In Chap e IV we decided o u he exploi he oppo uni ies p o ided by
SCNP owa ds pho oca aly ic applica ions, p o ing hei sui abili y as e icien
pho osensi ize nanoca ie s o pho odynamic he apy (PDT) applica ions. Fo his,
we de eloped a acile p o ocol o he encapsula ion o a syn hesized, long-wa eleng h
abso bing zinc(II)-ph halocyanine (ZnPc) wi hin he co e o a no el, sel -agg ega ing
gene a ion o amphiphilic copolyme s. Taking ad an age o he well-known sel -
assembly capabili y o an h acene molecules, we p epa ed an h acene-based
amphiphilic copolyme s bo h capable o sel -assembly in wa e and o e icien ly
encapsula ing he a in a ed- esponsi e complex ZnPc, yielding s able, wa e soluble,
ed-ligh eac i e SCNPs, as con i med by SAXS cha ac e iza ion. We hen es ed his
long-wa eleng h eac i e amphiphilic SCNPs agains human b eas cance cell MDA-
MB-231 lines o assess hei PDT e iciency. Ha ing obse ed ou s anding
46
pe o mance o one o he selec ed o mula ions, we inally es ed hei PDT ac i i y
in Zeb a ish emb yo xenog a s as a mo e accu a e human cance model.
Finally, he conclusions o he p esen hesis will be delinea ed in Chap e V.
47
1.6. Re e ences
(1) K aus, T.; Malaquin, L.; Schmid, H.; Riess, W.; Spence , N. D.; Wol ,
H. Nanopa icle p in ing wi h single-pa icle esolu ion. Na . Nano ech. 2007, 2, 570-
576.
(2) Lee, H.; Choi, T. K.; Cho, H. R.; Gha a i, R.; Wang, L.; Choi, H. J.;
Chung, T. D.; Lu, N.; Hyeon, T.; Choi, S. H.; Kim, D.-H. A g aphene-based
elec ochemical de ice wi h he mo esponsi e mic oneedles o diabe es moni o ing
and he apy. Na . Nano ech. 2016, 11, 566-572.
(3) Pa ka zidis, K.; Wang, H. S.; T uong, N. P.; Anas asaki, A. Chem
2020, 6, 1575-1588.
(4) Ni i, A.; Ca o a, R.; Assanelli, G.; No a i, M.; Pasini, D. Single-chain
polyme nanopa icles o add essing mo phologies and unc ions a he nanoscale: a
e iew. ACS Appl. Nano Ma e . 2022, 5, 10, 13985-13997.
(5) Ma yiaszewski, K.; Xia, J. A om ans e adical polyme iza ion.
Chem. Re . 2001, 101, 2921-2990.
(6) Hawke , C. J.; Bosman, A. W.; Ha h, E. New polyme syn hesis by
ni oxide media ed li ing adical polyme iza ions. Chem. Re . 2001, 101, 3661-3688.
(7) Moad, G.; Rizza do, E.; Thang, S. H. Li ing adical polyme iza ion by
he RAFT p ocess-a hi d upda e. Aus . J. Chem. 2012, 65, 985-1076.
(8) Moad, G.; Chie a i, J.; K s ina, J.; Pos ma, A.; Mayadunne, R. T. A.;
Rizza do, E.; Thang, S. H. Li ing ee adical polyme iza ion wi h e e sible addi ion
agmen a ion chain ans e ( he li e o RAFT). Polym. In . 2000, 49, 993–1001.
(9) Handbook o RAFT polyme iza ion. Edi ed by Ch is ophe Ba ne -
Kowollik. 2008 WILEY-VHC Ve lag GmbH & Co. KGaA, Weinheim.
(10) Be da, E. B.; Fos e , E. J.; Meije , E. W. Towa d con olling olding in
syn he ic polyme s: ab ica ing and cha ac e izing sup amolecula single-chain
nanopa icles. Mac omolecules 2010, 43, 1430-1437.
(11) Willcock, H.; O’Reilly, R. End g oup emo al and modi ica ion o
RAFT polyme s. Polym. Chem. 2010, 1, 149-157.
(12) a) A a , M.; Te ashima, T.; Sawamo o, M.; Meije , E. W.; palmans,
A. R. A. Unde s anding he ca aly ic ac i i y o single-chain polyme ic nanopa icles
in wa e . J. Polym. Sci. A Polym. Chem. 2014, 52, 12-20. b) Abdouni, Y.; e Huu ne,
G. M.; Yilmaz, G.; Monaco, A.; Redondo-Gómez, C.; Meije , E. W.; Palmans, A. R.
A.; Bece , C. R. Sel -assembled mul i- and single-chain glyconanopa icles and hei
lec in ecogni ion. Biomac omol. 2021, 22, 661-670. c) Liu, Y.; Tu unen, P.; de Waal,
48
B. F. M.; Blank, K. G.; Rowan, A. E.; Palmans, A. R. A.; Meije , E. W. Ca aly ic single-
chain polyme ic nanopa icles a wo k: om ensemble owa ds single-pa icle kine ics.
Mol. Sys . Des. Eng. 2018, 3, 609-618. d) Deng, L.; Albe azzi, L.; Palmans, A. R. A.
Elucida ing he s abili y o single-chain polyme ic nanopa icles in biological media
and li ing cells. Biomac omol. 2022, 23, 326-338.
(13) Zhang, C.; Yan, L.; Wang, X.; Zhu, S.; Chen, C.; Gu, Z.; Zhao, Y.
p og ess, challenges, and u u e o nanomedicine. Nano Today 2020, 35, 101008.
(14) Ha h, E.; Ho n, B. V.; Lee, V. Y.; Ge mack, D. S.; Gonzales, C. P.;
Mille , R. D.; Hawke , C. J. A acile app oach o a chi ec u ally de ined nanopa icles
ia in amolecula chain collapse. J. Am. Chem. Soc. 2002, 124, 8653-8660.
(15) Sanchez-Sanchez, A.; Ful on, D. A.; Pomposo, J. A. pH- esponsi e
single-chain polyme nanopa icles u ilizing dynamic co alen enamine bonds. Chem.
Commun. 2014, 50, 1871-1874.
(16) Beck, J. B.; Killops, K. L.; Kang, T.; Si anandan, K.; Bayles, A.;
Mackay, M. E.; Wooley, K. L.; Hawke , C. J. Facile p epa a ion o nanopa icles by
in amolecula c oss-linking o isocyana e unc ionalized copolyme s.
Mac omolecules 2009, 42, 5629-5635.
(17) K öge , A. P. P.; Hamelmann, N. M.; Juan, A.; Lindhoud, S.; Paulusse,
J. M. J. Biocompa ible single-chain polyme nanopa icles o d ug deli e y – a dual
app oach ACS Appl. Ma e . In e aces 2018, 10, 30946-30951.
(18) K öge , A. P. P.; Boonen, R. J. E. A.; Paulusse, J. M. J. Well-de ined
single-chain polyme nanopa icles ia hiol-Michael addi ion. Polyme 2017, 120,
119-128.
(19) K öge , A. P. P.; Paa s, W. D.; Boonen, R. J. E. A.; Hamelmann, N.
M.; Paulusse, J. M. J. Pen a luo ophenyl-based single-chain polyme nanopa icles as
a e sa ile pla o m owa ds p o ein mimic y. Polym. Chem. 2020, 11, 6056-6065.
(20) De Luzu iaga, A. R.; O ma egui, N.; G ande, H. J.; Od iozola, I.;
Pomposo, J. A.; Loinaz, I. In amolecula click cyccloaddi ion: an e icien oom-
empe a u e ou e owa ds bioconjugable polyme ic nanopa icles. Mac omol. Rapid
Commun. 2008, 29, 1156-1160.
(21) Chen, J.; Wang, J.; Li, K.; Wang, Y.; G uebele, M.; Fe guson, A. L.;
Zimme man, S. C. Polyme ic “clickase” accele a es he coppe click eac ion o small
molecules, p o eins, and cells. J. Am. Chem. Soc. 2019, 141, 9693-9700.
(22) Chen, J.; Li, K.; Bonson, S. E.; Zimme man, S. C. A bio hogonal
small molecule selec i e polyme ic “clickase” J. Am. Chem. Soc. 2020, 142, 13966-
13973.
(23) Maiz, J.; Ve de-Ses o, E.; Asenjo-Sanz, I.; Fouque , P.; Po ca , L.;
Pomposo, J. A.; de Molina, P. M.; A be, A.; Colmene o, J. Collec i e mo ions and
mechanical esponse o a bulk o single-chain nano-pa icles syn hesized by click-
chemis y. Polyme s, 2021, 13, 50.
49
(24) Willenbache , J.; Wues . K. N. R.; Muelle , J. O.; Kaupp, M.;
Wagenknech , A.; Ba ne -Kowollik, C. Pho ochemical design o unc ional luo escen
single-chain nanopa icles. ACS Mac o Le . 2014, 3, 574-579.
(25) Wues , K. N. R.; Lu, H.; Thomas, D. S.; Goldmann, A. S.; S enzel, M.
H.; Ba ne -Kowollik, C. Fluo escen glyco single-chain nanopa icle-deco a ed
nanodiamonds. ACS Mac o Le . 2017, 6, 1168-1174.
(26) O enloch, J. T.; Willenbache , J.; Tz e ko a, P.; Heile , C.; Mu lu, H.;
Ba ne -Kowollik, C. Deg adable, luo escen single-chain nanopa icles based on
me a hesis polyme s. Chem. Commun. 2017, 53, 775-778.
(27) O enloch, J. T.; Willenbache , J.; Tz e ko a, P.; Heile , C.; Mu lu, H.;
Ba ne -Kowollik, C. Deg adable, luo escen single-chain nanopa icles based on
me a hesis polyme s. Chem. Commun. 2017, 53, 775-778.
(28) Thannee u, S.; Li, W.; He, J.; Con ollable sel -assembly o
amphiphilic adpole-shaped polyme single-chain nanopa icles p epa ed h ough
in achain pho o-c oss-linking. Langmui , 2019, 35, 2619-2629.
(29) Maag, P. H.; Feis , F.; F isch, H.; Roesky, P. W.; Ba ne -Kowollik, C.
Fö s e esonance ene gy ans e wi hin songle chain nanopa icles. Chem. Sci. 2024,
15, 5218-5224.
(30) Blázquez-Ma ín, A.; Bona dd, S.; Ve de-Ses o, E.; A be, A.;
Pomposo, J. A. ACS Polym. Au 2024, 4, 140-148.
(31) Zhang, Y.; Xue, Y.; Gao, L.; Liao, R.; Wang, F.; Wang, F.; Me ging
non-co alen and co alen c osslinking: en ou e o single chain nanopa icles. CChem.
Le . 2024, 35, 109217.
(32) Li, X.; Han, M.; Zhang, H.; Liu, F.; Pan, Y.; Zhu, J.; Liao, Z.; Chen,
X.; Zhang, B. S uc u es and biological unc ions o zinc inge p o eins and hei oles
in hepa ocellula ca cinoma. Bioma ke Res. 2022, 10, 2.
(33) Knö el, N. D.; Ro h uss, H.; Tzue ko a, P.; Kulend an, B.; Ba ne -
Kowollik, C.; Roesky, P. W. He e obime alli Eu(III)/P (II) single-chain nanopa icles:
a pa h o enligh en ca aly ic eac ions. Chem. Sci. 2020, 11, 10331-10336.
(34) Bahley, J. L.; Kulend an, B.; Roesky, P. W.; He e obime allic
Au(I)/Y(II) single-chain nanopa icles as ecyclable homogeneous ca alys s. Polym.
Chem. 2021, 12, 4016-4021.
(35) Gillhube , S.; Halloway, J. O.; F isch, H.; Feis , F.; Weigend, F.;
Ba ne -Kowollik, C.; Roesky, P. W.; Fe ocene-d i en single-chain polyme
compac ion. Chem. Commun. 2023, 59, 4672-4675.
(36) Rei h, M. A.; Ka das, S.; Me ens, C.; Fossep é, M.; Su in, M.;
S einkoenig, J.; Du P ez, F. E.; Using nickel o old disc e e syn he ic mac omolecules
in o single-chain nanopa icles. Polym. Chem. 2021, 12, 4924-4933.
(37) Wang, W.; Wang, J.; Li, S.; Li, C.; Tan, R.; Yin, D. I on(II)- olded
single-chain nanopa icles: a me alloenzyme mimicking sus ainable ca alys s o highly
enan ioselec i e sul a-Michael addi ion in wa e . G een Chem. 2020, 22, 4645-4655.

50
(38) Mello, C. C.; Ba ick, D. An Expe imen ally de e mined p o ein
olding ene gy landscape. P oc. Na l. Acad. Sci. U. S. A. 2004, 101, 14102−14107.
(39) Seo, M.; Beck, B. J.; Paulusse, J. M. J.; Hawke , C. J.; Kim, S. Y.;
Polyme ic nanopa icles ia nonco alen c oss-linking o linea chains.
Mac omolecules 2008, 41, 6413-6418.
(40) Appel, E.A.; Dyson, J.; del Ba io, J.; Walsh, Z.; Sche man, O. A.
Fo ma ion o single-chain nanopa icles in wa e h ough hos -gues in e ac ions.
Angew. Chem. In . Ed. 2012, 51, 4185-4189.
(41) Ma sumo o, K.; Te ashima, T.; Sugi a, T.; Takenaka, M.; Sawamo o,
M. Amphiphilic andom copolyme s wi h hyd ophobic / hyd ogen-bonding u ea
pendan s: sel - olding polyme s in aqueous and o ganic media. Mac omolecules 2016,
49, 7917-7927.
(42) a) Hosono, N.; Gillissen, M. A. J.; Li, Y.; Sheiko, S. S.; Palmans, A.
R. A.; Meije , E. W.; o hogonal sel -assembly in olding block copolyme s. J. Am.
Chem. Soc. 2013, 135, 501-510. b) Fos e , E. J.; Be da, E. B.; Meije , E. W. Me as able
sup amolecula polyme nanopa icles ia in e molecula collapse o single polyme
chains. J. Am. Chem. Soc. 2009, 131, 6964-6966. c) Fos e , E. J.; Be da, E. B.; Meije ,
E. W. Tuning he size o sup amolecula single-chain polyme nanopa icles. J. Polym.
Sci. A Polym. Chem. 2011, 49, 118-126. d) Cheng, C.-C.; Chang, F.-C-; Yen, H.-C.;
Lee, D.-J.; Chiu, C.-W.; Xin, Z. Sup amolecula assembly media es he o ma ion o
single-chain polyme ic nanopa icles. ACS Mac o Le . 2015, 4, 1184-1188. e) Cheng,
C.-C.; Lee, D.-J.; Liao, Z.-S., Huang, J.-J. S imuli- esponsi e single-chain polyme ic
nanopa icles owa ds he de elopmen o e icien d ug deli e y sys ems. Polym.
Chem. 2016, 7, 6164-6169.
(43) Shao, Y.; Wang, Y.-L.; Tang, Z.; Wen, Z.; Chang, C.; Wang, C.; Sun,
D.; Ye, Y.; Qin, D.; Ke, Y.; Liu, F.; Yang, Z. Scalable syn hesis o pho oluminescen
single-chain nanopa icles by elec os a ic-media ed in amolecula c osslinking.
Angew. Chem. In . Ed. 2022, 61, e202205183.
(44) Guazzelli, E.; Maso i, E.; Calosi, M.; K iechbaum, M.; Chlig, F.;
Galli, G.; Ma inelli, E. Single-chain olding and sel -assembling o amphiphilic
polye hylene glycol-modi ied luo ina ed s y ene homopolyme s in wa e solu ion.
Polyme 2021, 231, 124107.
(45) Te ashima, T.; Sugi a, T.; Fukoa, K.; Sawamo o, M. Syn hesis and
single-chain olding o amphiphilic andom copolyme s in wa e . Mac omolecules
2014, 47, 589-600.
(46) Imai, S.; Hi ai, Y.; Nagao, C.; Sawamo o, M.; Te ashima, T.
P og ammed sel -assembly sys ems o amphiphilic andom copolyme s in o size-
con olled and he mo esponsi e micelles in wa e . Mac omolecules 2018, 51, 398-
409.
51
(47) Ve de-Ses o, E.; A be, A.; Mo eno, A. J.; Cangialosi, D.; Aleg ía, A.;
Colmene o, J.; Pomposo, J. A. Single-chain nanopa icles: oppo uni ies p o ided by
in e nal and ex e nal con inemen . Ma e . Ho iz. 2020, 7, 2292-2313.
(48) a) Single-Chain Polyme Nanopa icles: Syn hesis, Cha ac e iza ion,
Simula ions, and Applica ions, Edi ed by José Adol o Pomposo Wiley-VCH,
Weinheim, 2017. b) Ma ila, S.; Ei gi, O.; Be ko ich, I.; Lemco , N. G. In amolecula
c oss-linking me hodologies o he syn hesis o polyme nanopa icles. Chem. Re .
2016, 116, 878-961. c) Lyon, C. K.; P ashe , A.; Hanlon, A. M.; Tu en, B. T.; Tooley,
C. A.; F ank, P. G.; Be da, E. B. A b ie use ’s guide o single-chain nanopa icles.
Polym. Chem. 2015, 6, 181-197.
(49) a) Koda, Y.; Te ashima, T.; Sawamo o, M.; Mayna d, H. D.; P o ein
s o age wi h pe luo ina ed PEG compa men s in a hyd o luo oca bon sol en . Polym.
Chem. 2015, 6, 240-247. b) La o e-Sanchez, A.; Pomposo, J. A. Recen bioinspi ed
applica ions o single-chain nanopa icles. Polym. In . 2016, 65, 855-860.
(50) Robinson, P. K.; Enzymes p inciples and bio echnological
applica ions. Essays Biochem. 2015, 59, 1-41.
(51) Sahin, A.; Weiland , D. R.; Ha zimanika is, V.; Op imal enzyme
u iliza ion sugges s ha concen a ions and he modynamics de e mine binding
mechanisms and enzyme sa u a ions. Na . Commun. 2023, 16, 2618.
(52) Rubio-Ce illa, J.; González, E.; Pomposo, J. A.; Applica ions o
single-chain polyme nanopa icles. John Wiley & Sons.: Weinheim, Ge many 2017.
(53) Chen, J.; Li, K.; Bonson, S. E.; Zimme mann, S. C. A bio hogonal
small molecule selec i e polyme ic “clickase”. J. Am. Chem. Soc. 2020, 142, 13966-
13973.
(54) Chen, J.; Li, K.; Sean, J.; Shon, L.; Zimme mann, S. C.; Single-chain
nanopa icle deli e s a pa ne enzyme o concu en and andem ca alysis in cells. J.
Am. Chem. Soc. 2020, 142, 4565-4569.
(55) Mundsinge , K.; Tu en, B. T.; Wang, L.; Neubaue , K.; K op , C.;
O’Ma a, M. L.; Ba ne -Kowollik, C. Visible-Ligh -Reac i e Single-Chain
Nanopa icles Angew. Chem. In . Ed. 2023, 62, e202302995.
(56) Chen, X.; Chen, Z.; Ma, L. Mil i-s imuli- esponsi e bo leb ush-
colloid Janus nanopa icles owa d emulsion in e acial manipula ion and ca alysis.
Polym. Chem. 2022, 13, 959-966.
(57) Xu, W.; Ye, Y.; Sun, D.; Yang, Z. Single-chain nanopa icles ca alyzed
polyme iza ion owa d composi e nanopa icles. J. Polym. Scie. 2024, 62, 427-435.
(58) Hamelmann, N. M.; Paulusse, J. M. J. Single-chain polyme
nanopa icles in biomedical applica ions. J. Con ol. Release 2023, 356, 26-42.
(59) a) Liu, D.; He, C.; Wang, A. Z.; Lin, W. Applica ions o liposomal
echnologies o deli e y o pla inum analogs in oncology. In . J. Nanomed. 2013, 8,
3309-33149. b) Ko, A. H.; Tempe o, M. A.; Shan, Y. S.; Su, W. C.; Lin, Y. L.; Di o,
52
E.; Ong, A.; Wang, Y. W.; Yeh, C. G.; Chen, L. T. A mul ina ional phase 2 s udy o
nanoliposomal i ino ecan suc oso a e (PEP02, MM-398) o pa ien s wi h gemci abine-
e ac o y me as a ic panc ea ic cance . B i . J. Cance 2013, 109, 920–925. c)
Koudelka, S.; Tu ánek, J.; Liposomal pacli axel o mula ions. J. Con ol. Release
2012, 163, 322–334. d) Deeken, J. F.; Slack, R.; Weiss, G. J.; Ramana han, R. K.;
Pish aian, M. J.; Hwang, J.; Lewandowski, K.; Sub amanian, D.; He, A. R.; Co a la,
I.; A phase i s udy o liposomal-encapsula ed doce axel (LE-DT) in pa ien s wi h
ad anced solid umo malignancies. Cance Chemo h. Pha m. 2013, 71, 627-633.
(60) a) Song, C.; Lin, T.; Zhang, Q.; Thayumana an, S.; Ren, L. pH-
sensi i e mo phological ansi ions in polyme ic adpole assemblies o p og ammed
umo he apy. J. Con ol. Release 2019, 293, 1–9. b) Beni o, A. B.; Aie za, M. K.;
Ma adi, M.; Gil-Ice a, L.; Shekh e Zaha i, T.; Szczupak, B.; Jiménez-González, M.;
Reese, T.; Scanziani, E.; Passoni, L.; Ma eoli, M.; De Maglie, M.; O ens ein, A.; O on-
He man, M.; Kos enich, G.; Buzhansky, L.; Gazi , E.; G ande, H.-J.; Gomez-Vallejo,
V.; Llop, J.; Loinaz, I. Func ional single-chain polyme nanopa icles: a ge ing and
imaging panc ea ic umo s in i o, Biomac omolecules 2016, 17, 3213–3221.
(61) A ias-Alpiza , G.; Koch, B.; Hamelmann, N. M.; Neus up, M. A.;
Paulusse, J. M. J.; Jiskoo , W.; K os, A.; Bussmann, J. S abilin-1 is equi ed o he
endo helial clea ance o small anionic nanopa icles, Nanomed. Nano echnol. Biol.
Med. 2021, 34, 102395.
(62) a) Wues , K. N. R.; Lu, H.; Thomas, D. S.; Goldmann, A. S.; S enzel,
M. H.; Ba ne -Kowollik, C. Fluo escen glyco single-chain nanopa icle-deco a ed
nanodiamonds. ACS Mac o Le . 2017, 6, 1168–1174. b) K öge , A. P. P.; Komil, M.
I.; Hamelmann, N. M.; Juan, A.; S enzel, M. H.; Paulusse, J. M. J. Glucose single-chain
polyme nanopa icles o cellula a ge ing, ACS Mac o Le . 2019, 8, 95–101.
(63) Bajj, D. N. F; T an, M. V.; Tsai, H.-Y.; Kim, H.; Paisley, N. R.; Alga ,
W. R.; Hudson, Z. M. Fluo escen he e o elechelic single-chain polyme nanopa icles:
syn hesis, spec oscopy, and cellula imaging. ACS Appl. Nano Ma e 2019, 2, 898–
909.
(64) Tian, X.; Xue, R.; Yang, F.; Yin, L.; Luan, S.; Tang, H. Single-chain
nanopa icle-based coa ings wi h imp o ed bac e icidal ac i i y and an i ouling
p ope ies, Biomac omolecules 2021, 22, 4306–4315. b) Nguyen, ] T.-K.; Lam, S. J.;
Ho, K. K. K.; Kuma , N.; Qiao, G. G.; Egan, S.; Boye , C.; Wong, E. H. H. Ra ional
design o single-chain polyme ic nanopa icles ha kill plank onic and bio ilm bac e ia.
ACS In ec . Dis. 2017, 3, 237–248.
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60
In p esence o a hi d subs ance, he pho oca alys (PC), capable o abso b ligh
and o ake place in a ce ain p ocess wi hou being chemically al e ed, pho ochemical
ans o ma ion can occu e en be ween eac an s which ei he end no o eac o do
no eac a all he mally. In his case, he p ocess is said o be pho oca alyzed by PC.
F om he he modynamic poin o iew, a pho oca alyzed ans o ma ion can
occu ia wo di e en modali ies, which a e schema ized in Figu e 2.3: in he case b,
is desc ibed a pho osensi ized p ocess, while he case c is e e ed o he con e sion o
ligh in o chemical ene gy. In a pho osensi ized p ocess, a he modynamically allowed,
ye slow, eac ion be ween A and B o yield C and D, is sped up by he p esence o PC,
whose elec onic exci ed s a e gene a ed upon abso p ion o ligh augmen he o al ee
ene gy o he sys em. In he case o he con e sion o ligh in o chemical ene gy, an
ini ially he modynamically impossible eac ion becomes allowed by he p esence o
he pho oexci ed species PC*. Bo h modes o ac ion a e he esul an o he con ibu ion
o he elec onic exci ed s a e o PC o he o al ee Gibbs ene gy o he sys em.15,16
I is wo h no ing ha all pho oca alyzed eac ions can be g ouped in o wo
main classes, acco ding o he mechanism wi h which he PC enables he ca aly ic
cycle: (i) ene gy ans e and (ii) pho o edox ca alysis. In he eac ion d is schema ized
he main p ocess o pho osensi ized eac ions by ene gy ans e .
(d)
In ene gy ans e ca alysis, a iple exci ed s a e T1 o a molecule (3PC* o
he pho oca alys ) is deac i a ed o a lowe ene ge ic s a e by ans e ing ene gy o a
second molecule ( he subs a e A, he ene gy accep o ), which is he eby p omo ed o
a highe ene ge ic le el, usually i s iple exci ed s a e 3A*, wi h his la e , being
capable o decaying in o p oduc s (Figu e 2.4).

61
Figu e 2.4. Schema ic ep esen a ion (Jablonski diag am) o he main elec onic
ansi ions in ol ed in ene gy ans e pho oca aly ic p ocesses.17 The ine ib onic s uc u es
o he single s exci ed s a es 1Sn wi h in e nal elaxa ion he eo we e omi ed o pic o ial
cla i y. The eac i e iple s a e o he accep o subs a e 3A*, he c ucial in e media e in he
mechanis ic pa hway o p oduc s, is highligh ed in yellow. The spec oscopic iple ene gies
ET and he associa ed ansi ions a e ep esen ed in g ey.
2.1.2. Pho oca alysis o O ganic Reac ions.
As an icipa ed in he p e ious pa ag aphs, o access common o ganic
subs a es’ iple s a es h ough di ec exci a ion would gene ally equi e bo h sho
(280-315 nm) i adia ion wa eleng hs and high ligh in ensi ies, which esul in
unselec i e abso p ion and undesi ed side eac ions. E icien pho oca alys s, mus hen
exhibi high ex inc ion coe icien s and apid in e sys em c ossing (ISC), which in
conjunc ion wi h a long-li ing iple s a e (>100 ns), explains he p e alence o hea y-
a om Ru/I complexes and ca bonyl compounds as e icien iple sensi ize s.18
Pho ocycloaddi ions, he e oly ic bond dissocia ions, ca bon-ca bon double bonds
pho oisome iza ions and eac ions in ol ing he o ma ion o pho osensi ized single
62
oxygen a e he ou mos commonly used classes o eac ions in which he ene gy
ans e mechanism is in ol ed. In Figu e 2.5 a e shown some exempli ica i e
applica ions o he abo emen ioned ans o ma ions.
Figu e 2.5. On he le , ou examples o ecen ly epo ed pho oca alyzed eac ions
ia ene gy ans e mechanism: he he e oly ic bond dissocia ion o ca bazides,19 he
egioselec i e pho o Diels-Alde cycloaddi ions o an h acenes o ole ins,20 he single
oxygen cycloaddi ion o dienes,21 and he isome iza ion o ca bon-ca bon double bonds.22 On
he igh , he s uc u e o commonly used pho osensi ize s wi h hei espec i e ET (in
pa en heses) epo ed in kcal mol-1.18
Rega ding he second mode o ac ion o a pho oca alyzed eac ion, he
pho o edox ca alysis, he eac ion mechanism in ol es he elec on ans e be ween
he exci ed pho oca alys and subs a es, acili a ed by he p inciple discussed abo e by
which he exci ed s a es o a pho oca alys a e usually bo h be e educ an s and be e
oxidize s.15 I PC’s exci ed s a e ans e s an elec on o one subs a e o ini ia e he
ca aly ic cycle, he mechanis ic ou e is called educ i e quenching cycle (Scheme 2.1),
which is usually comple ed by he educ ion o he oxidized in e media e o he
pho oca alys ia elec on ans e o m a second, elec on dona ing eagen . In some
cases, he edox po en ials o he species in ol ed in he cycle can unde go he opposi e
p ocess, an oxida i e quenching cycle, in which he pho oexci ed s a e o PC abs ac
an elec on om a dono o ini ia e he ca alysis (Scheme 2.1).
63
Scheme 2.1. Possible mechanisms cycles allowed o pho o edox ca alysis. On he
igh he pho oca alys PC is educed by a dono eac an D ia single-elec on ans e (SET)
a e being exci ed upon ligh abso p ion, he educed o m o PC is oxidized by an accep o
eac an A o inally yield he egene a ed PC. On he le , a e being p omo ed o he eac i e
exci ed s a e upon ligh abso p ion, PC* ans e s an elec on ia SET o an accep ing eac an
A. The g ound s a e pho oca alys is hen egene a ed by a second SET s ep in which a
dona ing eac an D is oxidized by he in e media e oxidized PC.
In he case o he mally-equilib a ed exci ed s a es and e e sible elec on
ans e , he edox po en ials o he eac i e exci ed s a e o PC (PC*) can be calcula ed
acco ding o Equa ions 1 and 2.
E edox(PC+/PC*) = E edox(PC+/PC) – Eoo(PC*/PC)
Eq. (1)
E edox(PC*/PC-) = E edox(PC/PC-) + Eoo(PC*/PC)
Eq. (2)
Whe e Eoo(PC*/PC) is he one-elec on po en ial co esponding o he ze o-
ze o spec oscopic ene gy o he exci ed s a e and E edox(PC/PC-) and E edox(PC+/PC) he
g ound s a e edox po en ials measu ed o PC.23 As a gene al ule, i is possible o une
64
he oxida ion po en ials o he pho oca alys o swi ch om one mechanism o ano he ,
opening new syn he ically aluable possibili ies.24
Pho o edox ca alysis, and isible-ligh pho o edox ca alysis in pa icula , is an
undoub edly powe ul, ye s ill lou ishing app oach o he p epa a ion o o ganic
molecules. Fi s pionee ing epo s o pho o edox ca alysis we e only published
s a ing o m 1979 by Kellogg,25 ollowed by Fukuzumi and Tanaka,26 Pac and
De onzie ,27 all conce ning he capabili y o u henium polypy idyl complexes o
accele a e a a ie y o o ganic eac ion, in Figu e 2.6 some signi ica i e examples a e
shown.
Figu e 2.6. Th ee ep esen a i e examples o his o ically ele an pho o edox
o ganic eac ion. F om he op: he ligh -induced accele a ion o he educ i e desul u a ion
o ialkyl sul onium sal s in p esence o ca aly ic amoun s o [Ru(bpy)3]Cl2, epo ed by
Kellogg in 1979.25 The ligh -dependen educ i e coupling o benzyl b omide in p esence o
1-benzyl-1,4-dihyd onico inamide and ca aly ic amoun o [Ru(bpy)3]Cl2.26 The pho o-
oxida ion o benzylic alcohols o he co esponding aldehydes, media ed by polypy idyl
Ru(II) complexes and using diazonium sal s as sac i icial elec on dono s.27
By he beginning o his cen u y, pho o edox ca alysis s a ed i s blossoming
season wi h he wo ks o he Nobel Lau ea e MacMillan and Yoon’s g oups on he
65
e ec o isible ligh on o ganic eac ions in p esence o me al complexes.28 F om hen
on, pho o edox ca alysis now coun s wi h a ple ho a o di e en applica ions and a
a ie y o eac ions, o en ca ied ou in milde condi ions han hei he mal analogues,
ha e been disco e ed and op imized.15 Due o he possibili y o in oduce s uc u ally
complex moie ies in mild condi ions, an inc easing numbe o au ho i a i e indus ies
decide o in oduce pho o edox ca alysis-based eac ions in he syn hesis and la e-s age
unc ionaliza ion o highly- aluable molecules.29
Rega dless he applica ion, mos ly all pho ochemis y and pho oca alysis-
based echnologies ha e exponen ially g own du ing he cou se o he pas decade, a
ise in de elopmen which can be easily co ela ed o he a ailabili y o no el, mo e
s able and powe ul ligh sou ces. Especially, ela ed o expe imen al se -ups o
isible-ligh pho oca alysis p ac ice, which adi ionally elied on he use o ei he he
sunligh o household ligh bulbs, he comme cializa ion o s anda dized ligh emi ing
diodes (LED) pe mi ed a ne gain in e ms o eac ion epea abili y (LEDs sou ces
usually p esen na ow spec al emission) and o e all pe o mance.30 I is also
impo an o ake in mind ha ligh is always a p ope eac an in pho oca alyzed
p ocesses, which is s oichiome ically consumed, hough aking pa nea ly in he inal
yield o a p ocess and making i s ic ly dependen o he ligh in ensi y o he sou ce
employed.30,31 The endency can be isually app ecia ed in Figu e 2.7, in which a
g aphic o he es ima ion o he numbe o publica ions wi h ime in he ields o isible-
ligh pho oca alysis and pho ochemis y is epo ed and supe imposed wi h h ee o he
miles one op oelec onic in en ions, which undoub edly con ibu ed o he e olu ion
o he scien i ic communi y in he pas decade.32

66
Figu e 2.7. G aphic o he endencies, gi en by he numbe o publica ions
con aining he exp essions isible-ligh pho oca alysis, pho ochem and pho oca alysis pe
quinquennia, s a ing om 1910s o 2020s and de i ed om he digi al sea ch po al Google
Schola (14 h Feb ua y 2024).
2.2. Discussion
The ollowing discussion will mainly ocus on ecen de elopmen s o so -
ma e based pho oca aly ic sys ems, capable o ca ying ou o ganic eac ions in wa e .
The use o sup amolecula sca olds is one o he possible s a egies explo ed by ecen
de elopmen in he ield o pho oca alysis o add ess, no only he majo cons ain s o
sca ce solubili y o ca alys s and p oduc s, bu which is capable o ou pe o m na u al
sys ems in many cases. In sup amolecula sys ems, disc e e molecules in e ac and
come oge he o o m assembled en i ies. This app oach, gene ally inspi ed by
biological eac ions which o en occu in he con ined pocke s o enzymes, allow
lowe ing he ansi ion s a e ene gies o eac an molecules, which a e hos ed in he
sup amolecula assemblies’ nanoca i ies and a e acili a ed o eac in he
mic oen i onmen p o ided by he con inemen .33,34 A a ie y o sup amolecula hos s
ha e been de eloped wi h well-de ined nanoca i ies o mimic enzyme ac i i y,
scien i ic li e a u e is plen y o b illian examples o how sup amolecula chemis s a e
67
capable o employ weak, nonco alen in e ac ions o build de ined and disc e e
molecula a chi ec u es o ca alysis pu poses35-39 and, no ably, sup amolecula hos s
a e, in some cases, supe io o molecula ca alys s in e ms o highe yields and be e
selec i i y.40-42 Ne e heless, he applica ion o sup amolecula sys ems o isible-
ligh pho oca alysis o o ganic ans o ma ions in wa e s ill is in i s in ancy and,
al hough some ep esen a i e examples epo ed in li e a u e al eady disclosed he g ea
po en ial o his app oach, ques ions and unsol ed majo issues emain abundan .
2.2.1. Wa e as Sol en in O ganic Pho oca alysis.
One could easily and easonably a i m ha , when hinking abou
pho oca alysis, scien is s a e someway aking o ook inspi a ion om Na u e.
Conside ing he cen al ole ha he pho oca aly ic p ocess plays in he ecological
homeos asis o li e, he plane ea h i sel can be conside ed as a big, almos inde ini ely
complex pho o eac o . In 1772, Joseph P ies ley epo ed he i s expe imen
demons a ing he p oduc ion o oxygen gas by plan s, while i e yea s la e Jan Ingen-
Housz documen ed ha he p oduc ion o oxygen gas in plan s lea es was a ligh -
dependen p ocess.43 F om hen on, coun less o scien is s s a ed he ace owa ds he
comp ehension o he phenomenon, which emained co e ed in mys e y up un il he
ad en o cell biochemis y and mode n biomolecula echniques. In his scena io, i s
epo s o pho oca alyzed eac ions in wa e appea ed. P obably aiming o simula e and
un eil he p ocesses behind plan s’ pho osyn hesis, Ba ke epo ed in 1921 he pho o-
induced in wa e syn hesis o o maldehyde and ca bohyd a es om ca bon dioxide in
p esence o u anium sal s and colloidal i on oxides.44 Apa om some mo e seminal
epo s conce ning zinc and i anium oxides pho obleaching p ope ies in ae obic
en i onmen ,45 pho oca alysis in aqueous media la gely emained an uncha ed
challenge un il he las decade.
Ne e heless, Wohle ’s syn hesis o u ea, pe o med by hea ing an aqueous
solu ion o ammonium cyana e, was de eloped by he i s mid o he 19 h cen u y and
is commonly conside ed he beginning o syn he ic o ganic chemis y. As well as o
68
many o he ans o ma ions epo ed in he same pe iod, he use o wa e as sol en o
eac ion op imiza ion was no uncommon up un il he second decade o he 20 h
cen u y,46 a endency which ha e been su ely in e ed wi h he ad en o G igna d
eac an s and he ise o pe ochemical indus y.28,47 I should be poin ed ou ha
eplacing haza dous sol en s wi h sa e , enewable al e na i es is ecei ing inc easing
a en ion bo h in academia and indus ies. Almos all o ganic sol en s, excep
chlo ina ed sol en s, a e lammable, chlo ina ed and a oma ic sol en s a e
ca cinogenic, e he and chlo o o m ha e na co ic p ope ies, high apo p essu es and
o m smog, jus o ci e ew o hei e y undesi able cha ac e is ics.48 Making a p ocess
g eene , he e o e dec easing he en i onmen al impac and isks associa ed wi h
handling and esidues managemen , is undoub edly linked o he use o g eene sol en s
al e na i es. The ac ha wa e ul ills all he c i e ia o being an excellen ly g een
sol en is commonly ag eed, as i is no only allowing he minimiza ion o heal h and
en i onmen al isks, bu also p esen s unique oppo uni y o p oducing pola i y-
unable eac ion media.49,50
In addi ion, when wa e is used as sol en , di e en and unexpec ed eac i i ies
can be disclosed. Due o his enewed a en ion o sus ainabili y, i seems logical o
me ge he in insically g een ea u es o pho oca alysis o he bene i s ha he use o
wa e as eac ion media could p o ide. As highligh ed by ecen e iews on he opic,
he same can be said o pho oca alysis, s ill li le ha e been epo ed on pu ely in wa e
pho oca alyzed o ganic eac ions,28,51 as he sca ce solubili y o he mos po en
pho oca alys s and eac an s is o majo conce n. In Figu e 2.8, some signi ica i e
examples o pho oca aly ic syn hesis o o ganic molecules in which wa e is used as
he sole sol en medium a e schema ized.
69
Figu e 2.8. Six ep esen a i e examples o ligh -induced pho oca aly ic o ganic
eac ions allowed in wa e : a) A yla ion o N-he e oa enes wi h a yldiazonium sal s, i s
epo ed in 2014 by Xue e al.52 b) Visible-ligh -p omo ed oxida i e adical cycliza ion o N-
N-bia ylglycines, epo ed by Na a ajan e al. in 2019.53 c) In wa e , isible ligh -induced
acyla i e epoxida ion o inyla enes, epo ed by Salles e al. in 2018.54 d) A oma i e
dehyd ogena ion o cyclic amines, i s epo ed by Bala aman’s g oup in 2019.53 e) Visible-
76
2.2.3. Sup amolecula Nanocapsules.
While explo ing he di e en possibili ies o e ed by sup amolecula
in e ac ions o pho oca alysis in wa e , i is wo h epo ing he use o ailo -made so
nanocapsules o he s abiliza ion o hyd ophobic pho oca alys in wa e . This s a egy
was ema kably employed by Aki a e al. by p epa ing V-shaped a oma ic amphiphiles,
whose s uc u e is shown in Figu e 2.12, which a e capable o spon aneously sel -
assemble in wa e and gene a e hyd ophobic nanoca i ies, subsequen ly exploi ed o
hos he phenoxazine pho o edox ca alys PN.71 The au ho s epo ed ha , he esul ing
sup amolecula assembly (SAPC, Figu e 2.12), composed o o ganic PC enclosed in
he V-shaped a oma ic amphiphile, esul ed o e icien ly ca alyze he me al- ee
pinacol coupling in wa e , using blue LED ligh as he only ex e nal ene gy sou ce. The
same concep was success ully applied by he same g oup, which exploi ed he same
o ganic V-shaped amphiphile o ap h ee di e en pho oca alys PC (Scheme 2.3),
enabling a deme hoxyla i e educ i e clea age o N-O bonds in Wen eib amides (WA,
Scheme 2.3) in wa e , mild condi ions and good yields on a easonably la ge subs a e
pool (62 o 88% yield).72
Figu e 2.12. Pic o ial ep esen a ion o he in wa e sel -assembly o he
nanoca i ies p epa ed by Aki a e al. The V-shaped o ganic amphiphile spon aneously
agg ega e in aqueous en i onmen , gene a ing hyd ophobic nanoca i ies in which lipophilic
pho oca alys as PC can be hos ed.71

77
Scheme 2.3. On he le , s uc u es o he h ee di e en o ganic pho o edox
ca alys s selec ed by Aki a e al. o he inco po a ion wi hin he sup amolecula assembly
(SAPC). On he igh , eac ion scheme o he in wa e educ i e clea age o N-O bond o
WA.72
2.2.4. So Polyme ic Ma e ials o Aqueous O ganic Pho oca alysis.
As an icipa ed abo e, in all hese wo ks in he ield o pho oca alysis o o ganic
ans o ma ions in wa e , we obse ed he ex ensi e use o sup amolecula assembly
o simul aneously shield he pho oca aly ic species om he aqueous media and o
p o ide a con ined en i onmen in which he subs a es can be hos ed and unde go he
desi ed p ocess. While polyme ic sca old has been al eady widely employed o
ca alys suppo ing, demons a ing hei supe io i y in e ms o obus ness, ca alys
ecyclabili y, p oduc selec i i y enhancemen and es ic ed ca alys leaching, s ill
li le ha e been desc ibed on hei applica ion in pho oca alysis. Poly inylpy olidone
(PVP)-s abilized colloidal pla inum nanopa icles in combina ion wi h a wa e -soluble
zinc po phy in we e exploi ed o p omo e he isible-ligh educ ion o he subs a e
py u a e o lac a e, enabling a pho o edox-based, mild and g een p ocedu e o he
p epa a ion o a aluable aw ma e ial o he p epa a ion o biodeg adable polyme ic
compounds.73
Palmans e al. epo ed wo seminal wo ks in which wa e -soluble pho oac i e
SCNPs we e p o en o be capable o e icien ly ca alyze he me al- ee educ ion and
a oma ic C-C c oss-coupling eac ions in wa e and wi hou he use o any
cosol en .74,75 In hei epo s, he au ho s in oduce a sys ema ic me hodology o he
p epa a ion o s able, sel -assembled pho oca aly ic SCNPs o de ined s uc u e and
chemical composi ion. In hei i s wo k, a high-molecula weigh , poly(pen a luo o
phenyl) ac yla e p ecu so was modi ied h ough a pos - unc ionaliza ion app oach
78
wi h ou di e en unc ionali ies including pheno hiazine pendan s, ensu ing bo h he
homogenei y o he molecula weigh o he polyme ic p ecu so s and o con e he
pho o edox p ope ies o he ma e ial (Figu e 2.13). The ob ained amphiphilic
copolyme s esul ed o spon aneously sel -assemble in wa e o yield well-de ined non-
co alen SCNPs, which we e exploi ed o pe o m he UV-ligh induced educ ion o
haloa enes and he c oss-coupling o 2-cyanoiodobenzene and N-me hyl py ole in
wa e and in p esence ie hylamine (Scheme 2.4).74
Figu e 2.13. On op, he o ma ion o non-co alen , pho oca aly ic SCNP upon
spon aneous olding/collapse in wa e is depic ed. The unimolecula polyme ic agg ega e
enables he pho oca aly ic p ocess o occu in i s lipophilic, con ined co e. On he bo om,
he syn he ic ou e explo ed by Palmans e al. o he p epa a ion o he amphiphilic
pho o edox SCNP polyme ic p ecu so is shown.74
79
Scheme 2.4. Reac ion schemes o he “in wa e ”, ligh -d i en ans o ma ion
explo ed by Palmans’ g oup in hei wo ks on pho oca aly ic SCNPs (PC-SCNPs).74,75 On
op, he educ ion o C-halogen bonds o a oma ic subs a es, on he bo om he c oss-
coupling o N-me hyl py olidone and iodoa enes.
Ba ne -Kowollik e al. ecen ly epo ed a SCNPs-based pho o eac o capable
o pe o ming pho ooxida ion o a y acid in wa e and unde isible-ligh i adia ion.76
No ably, h ough he p ecise con ol o he olding/collapse p ocess o he polyme ic
chains by means o he use o a ose Bengal de i a i e c osslinke , he au ho s we e
able o une he deg ee o hyd ophobici y o he lipophilic pocke s wi hin he co e o
he single-chain nano eac o . In e es ingly, he s iking augmen in e iciency (up o
h ee- imes) o he pho oca aly ic SCNP owa d he pho ooxida ion o a y acids in
compa ison o he ee-pho osensi ize was in e p e ed, and suppo ed by MD
simula ions, in e ms o augmen ed pola i y and high con inemen in he ac i e pocke s
p o ided by he SCNP.76
2.3. Conclusions.
To summa ize, despi e he g ea in e es eme ged amongs pu e o ganic
chemis s owa ds isible-ligh pho oca alysis in he las decades and conside ing he
quick las cen u y ise o pho ochemis y, he implemen a ion o pho oca alysis and
80
so -ma e based sys ems o pe o ming aluable and challenging o ganic eac ion
s ill is in i s in ancy. Wi h his b ie o e iew, is in he au ho ’s hope ha he
impo ance, bo h academically and ecologically speaking, ha e been s essed ou . In
he nex Chap e s, we will y o u he ly highligh he po en ial o me ging
pho oca alysis and so -ma e sys ems o in wa e applica ions, by means o e acing
dome ema kable esul s in he ields o pho oca alysis in wa e and nanomedicine.
81
2.4. Re e ences.
(1) “pho oca alysis.” Collinsdic iona y.com. 2024.
h ps://www.collinsdic iona y.com/dic iona y/english/pho oca alysis (06 Feb ua y
2024).
(2) “pho oca alysis.” Oed.com. 2024.
h ps://www.oed.com/dic iona y/pho oca alysis_n (06 Feb ua y 2024).
(3) “pho oca alysis.” Me iam-webs e .com. 2024. h ps://www.me iam-
webs e .com/medical/pho oca alysis (06 Feb ua y 2024).
(4) “pho oca alysis” in IUPAC Compendium o Chemical Te minology,
3 d ed. In e na ional Union o Pu e and Applied Chemis y; 2006. Online e sion 3.0.1,
2019. h ps://doi.o g/10.1351/goldbook.P04580.
(5) Some expe imen s on he combus ion o he diamond and o he
ca bonaceous subs ances by Si Humph y Da y Phil. T ans. R.S. 1814, 104, 557-570.
(6) Ma in, T. The ins umen used by Da y and Fa aday in Flo ence o
he combus ion o diamonds. J. Sci. Ins um. 1931. 8 379.
(7) h ps://ca alogue.museogalileo.i /objec /Lens.h ml (06 Feb ua y
2024).
(8) “pho ochemis y” in IUPAC Compendium o Chemical Te minology,
3 d ed. In e na ional Union o Pu e and Applied Chemis y; 2006. Online e sion 3.0.1,
2019. h ps://doi.o g/10.1351/goldbook.P04588.
(9) T ommsdo , H. Uebe San onin. Annalen de Pha macie 1834, 11,
190-207.
(10) a) Ma suu a, T.; Sa o, Y.; Ogo o, R. A no el pho o ea angemen o
san onin in he solid s a e. Te ahed on Le . 1968, 44, 4627-4630. b) Fisch, M. H.;
Richa ds, J. H. The mechanism o he pho ocon e sion o san onin. J. Am. Chem. Soc.
1963, 85, 3029–3030. c) Chen, X.; Rinke icius, Z.; Luo, Y.; Åg en, H.; Cao, Z. The
mechanism o he pho ocon e sion o san onin. ChemPhysChem 2012, 13, 353 – 362.
(11) a) Becque el, E. Mémoi e su les e e s élec iques p odui s sous
l'in luence des ayons solai es. Comp es Rendus 1839, 9, 561–567. b) Becque el, E.
Mémoi e su le ayonnemen chimique qui accompagne la lumiè e solai e e la lumiè e
élec ique. Comp es Rendus 1840, 11, 702-703.
(12) a) B une , L.; Kozak, Z. Elek ochem. Angew. Phys. Chem. 1911, 17,
354. c) Eibne , A. Chem.-Z g. 1911, 35, 753. b) Eibne , A. Chem.-Z g. 1911, 35, 774.

82
(13) Ciamician, G. The pho ochemis y o he u u e. Science 1912, 36, 385-
394.
(14) a) Allmand, A. J. Pho ochemis y, 1914-1925. Annu. Rep. P og.
Chem., 1925, 22, 333-373.
(15) Megan, H.; Twil on, J.; MacMillan, D. W. C. Pho o edox ca alysis in
o ganic chemis y. J. O g. Chem. 2016, 81, 6898−692.
(16) Kozlowksi, M.; Yoon, T. Edi o ial o he special issue on
pho oca alysis. J. O g. Chem. 2016, 81, 6895−6897.
(17) Du a, S.; E chinge , J., E.; S ie h-Kal ho , F.; Kleinmans, R.;
Glo ius, F. Ene gy ans e pho oca alysis: exci ing modes o eac i i y. Chem. Soc.
Re ., 2024, 53, 1068-1089.
(18) S ie h-Kal ho , F.; James, M. J.; Tede s, M.; Pi ze , L.; Glo ius, F.
Ene gy ans e ca alysis media ed by isible ligh : p inciples, applica ions, di ec ions.
Chem. Soc. Re . 2018, 47, 7190-7202.
(19) Fa ney, E. P.; Yoon, T. Visible-ligh sensi iza ion o inyl azides by
ansi ion-me al pho oca alysis. Angew. Chem. In . Ed. 2014, 53, 793-797.
(20) Zhou, C.; Liu, Z.; Liang, Y.-Q.; Lei, T.; Chen, B.; Liao, R.-Z.; Tung,
C.-H.; Wu, L.-Z. Regioselec i e Diels–Alde eac ions o an h acenes wi h ole ins ia
isible ligh pho oca alysis in a homogeneous solu ion. O g. Le . 2024, 26, 1116-1121.
(21) a) Ghoga e, A. A.; G ee , A. Using single oxygen o syn hesize na u al
p oduc s and d ugs. Chem. Re . 2016, 116, 17, 9994–10034. b) Jung, M.; Ham, J.;
Song, J. Fi s To al Syn hesis o Na u al 6-Epiplako olide E. O g. Le . 2002, 4,
2763−2765.
(22) Me e nich, J. B.; A iukhin, D. G.; Holland, M. C.; on B emen-
Kuhne, M.; Neugebaue , J.; Gilmou , R. Pho oca aly ic E → Z isome iza ion o
pola ized alkenes inspi ed by he isual cycle: mechanis ic dicho omy and o igin o
selec i i y. J. O g. Chem., 2017, 82, 9955–9977.
(23) Balzani, V.; Be gamini, G.; Ce oni, P. Pho ochemis y and
pho oca alysis. Rend. Fis. Acc. Lincei 2017, 28, 125–142.
(24) Teega din, K.; Day, J. I.; Chan, J.; Wea e , J. Ad ances in
pho oca alysis: a mic o e iew o isible ligh media ed u henium and i idium
ca alyzed o ganic ans o ma ions. O g. P ocess Res. De . 2016, 20, 1156−1163.
(25) an Be gen, T. J.; Heds and, D. M.; K uizinga, W. H.; Kellogg, R. M.
Chemis y o dihyd opy idines. 9. Hyd ide ans e om 1,4-dihyd opy idines o sp3-
83
hyb idized ca bon in sul onium sal s and ac i a ed halides. S udies wi h NAD(P)H
models. J. O g. Chem. 1979, 44, 4953−4962.
(26) Hi onaka, K.; Fukuzumi, S.; Tanaka, T. T is(bipy idyl) u henium(II)-
pho osensi ized eac ion o 1-benzyl-1,4-dihyd onico inamide wi h benzyl b omide. J.
Chem. Soc., Pe kin T ans. 2, 1984, 1705-1709.
(27) Cano-Yelo, H.; De onzie , A. Pho o-oxida ion o some ca binols by
he Ru(II) polypy idyl complex-a yl diazonium sal sys em. Te ahed on Le . 1984,
25, 5517−5520.
(28) Russo, C.; B unelli, F.; T on, G. C.; Gius iniano, M. Visible-ligh
pho o edox ca alysis in wa e . J. O g. Chem. 2023, 88, 6284−6293.
(29) Candish, E.; Collins, K. D.; Cook, G. C.; Douglas, J. J.; Gómez-Suá ez,
A.; Joli , A.; Keess, S. Pho oca alysis in he li e science indus y. Chem. Re . 2022,
122, 2907-2080.
(30) Gesmundo, N. J.; Shaw, M. H.; Twil on, J.; Tellis, J. C.; MacMillan D.
W. C.; Nicewicz, D. A. Pho o edox Ca alysis In oduc ion, Desk Re e ence, and Use ’s
Guide. Me ck KGaA 2019
(31) Buglioni, L.; Raymenan s, F.; Sla e y, A.; Zondag, S. D. A.; Noël T.
Technological inno a ions in pho ochemis y o o ganic syn hesis: low chemis y,
high- h oughpu expe imen a ion, scale-up, and pho oelec ochemis y. Chem. Re .
2022, 122, 2752–2906.
(32) Zhelude , N. The li e and imes o he LED — a 100-yea his o y.
Na u e Pho on. 2007, 1, 189-192.
(33) Valla oju, N.; Si agu u, J., Sup amolecula pho oca alysis: combining
con inemen and non-co alen in e ac ions o con ol ligh ini ia ed eac ions. Chem.
Soc. Re . 2014, 43, 4084-4101.
(34) Raynal, M.; Balles e , P.; Vidal-Fe an, A.; an Leeuwen, P. W. N. M.,
Sup amolecula ca alysis. Pa 1: non-co alen in e ac ions as a ool o building and
modi ying homogeneous ca alys s. Chem. Soc. Re . 2014, 43 (5), 1660-1733.
(35) Wies e , M. J.; Ulmann, P. A.; Mi kin, C. A. Enzyme mimics based
upon sup amolecula coo dina ion chemis y. Angew. Chem., In . Ed. 2011, 50,
114−137.
(36) Koblenz, T. S.; Wassenaa , J.; Reek, J. N. H. Reac i i y wi hin a
con ined sel -assembled nanospace. Chem. Soc. Re . 2008, 37, 247−262.
84
(37) B own, C. J.; Tos e, F. D.; Be gman, R. G.; Raymond, K. N.
Sup amolecula ca alysis in me al−ligand clus e hos s. Chem. Re . 2015, 115,
3012−3035.
(38) Meeuwissen, J.; Reek, J. N. H. Sup amolecula ca alysis beyond
enzyme mimics. Na . Chem. 2010, 2, 615−621.
(39) Yoshizawa, M.; Klos e man, J. K.; Fuji a, M. Func ional molecula
lasks: new p ope ies and eac ions wi hin disc e e, sel -assembled hos s. Angew.
Chem., In . Ed. 2009, 48, 3418−3438.
(40) Mo imo o, M.; Bie schenk, S. M.; Xia, K. T.; Be gman, R. G.;
Raymond, K. N.; Tos e, F. D. Ad ances in sup amolecula hos -media ed eac i i y.
Na . Ca al. 2020, 3, 969−984.
(41) Yoshizawa, M.; Tamu a, M.; Fuji a, M. Diels-Alde in aqueous
molecula hos s: unusual egioselec i i y and e icien ca alysis. Science 2006, 312,
251−254.
(42) Zhang, Q.; Ca i, L.; Pleiss, J.; Tie enbache , K. Te pene cycliza ions
inside a sup amolecula ca alys : lea ing-g oup-con olled p oduc selec i i y and
mechanis ic s udies. J. Am. Chem. Soc. 2017,139,11482−11492.
(43) Go indjee, J. T. B.; Ges , H.; Allen, J. F.; Disco e ies in
pho osyn hesis. Ad ances in Pho osyn hesis and Respi a ion, 2005.
(44) a) Baly, E. C. C.; Heilb on, I. M.; Ba ke , W. F.; CX.—Pho oca alysis.
Pa I. The syn hesis o o maldehyde and ca bohyd a es om ca bon dioxide and
wa e . J. Chem. Soc., T ans. 1921, 119, 1025-1035. b) Baly, E.C.C.; Heilb on, I. M.;
S e n, H.J. XXIII.—Pho oca alysis. Pa III. The pho osyn hesis o na u ally occu ing
ni ogen compounds om ca bon dioxide and ammonia. J. Chem. Soc., T ans. 1923,
123, 185-197.
(45) Co onado, J. M.; F esno, F.; He nández-Alonso, M. D.; Po ela, R.
Design o ad anced pho oca aly ic ma e ials o ene gy and en i onmen al
applica ions. G een Ene gy and Technology, 2013.
(46) (a) Kobayashi, S. Science o Syn hesis: Wa e in O ganic Syn hesis.
Thieme Chemis y, 2012. (b) Linds om, U. M. O ganic Reac ions in Wa e ; Blackwell:
Ox o d, 2007.
(47) Ki anosono, T.; Masuda, K.; Xu, P.; Kobayashi, S. Ca aly ic o ganic
eac ions in wa e owa d sus ainable socie y. Chem. Re . 2018, 118, 679-746.
85
(48) By ne, F. P.; Jin, S.; Giulia, P.; Pe chey, T. H. M.; Cla k, J. h.; Fa me ,
J. T.; Hun , A. J.; McEl oy, C. R.; She wood, J. Tools and echniques o sol en
selec ion: g een sol en selec ion guides. Sus. Chem. P oc. 2016, 4, 7.
(49) Me ce , S. M.; Jessop, P. G. “Swi chable wa e ”: aqueous solu ions o
swi chable ionic s eng h. ChemSusChem 2010, 3, 467-470.
(50) Me ce , S. M.; Robe , T.; Dixon, D. V.; Chen, C.-S.; Ghoshouni, Z.;
Ha jani, J. R.; Jahangi i, S.; Peslhe be, G. H.; Jessop P. G. Design, syn hesis, and
solu ion beha io o small polyamines as swi chable wa e addi i es. G een Chem.,
2012, 14, 832-839.
(51) Hao, Y.; Lu, Y.-L.; Jiao, Z.; Su, C.-Y. Pho oca alysis mee s
con inemen : an eme ging oppo uni y o pho oinduced o ganic ans o ma ions.
Angew. Chem. In . Ed. 2024, DOI: 10.1002/anie.202317808.
(52) Xue, D.; Jia, Z.-H.; Zhao, C.-J.; Zhang, Y.-Y.; Wang, C.; Xiao, J.
Di ec a yla ion o N-he e oa enes wi h a yldiazonium sal s by pho o edox ca alysis in
wa e . Chem. Eu . J. 2014, 20, 2960 – 2965.
(53) Na a ajan, P.; Chuski , D.; P iya, S. Me al- ee, isible-ligh -p omo ed
oxida i e adical cycliza ion o N-bia ylglycine es e s: one-po cons uc ion o
phenan h idine-6-ca boxyla es in wa e . G een Chem., 2019, 21, 4406-4411.
(54) De Souza, G. F. P.; Bonacin, J. A.; Salles J , A. G. Visible-ligh -d i en
epoxyacyla ion and hyd oacyla ion o ole ins using me hylene blue/pe sul a e sys em
in wa e . J. O g. Chem. 2018, 83, 8331-8340.
(55) Ke zig, C.; Wenge , O. S.; Reac i i y con ol o a pho oca aly ic
sys em by changing he ligh in ensi y. Chem Sci. 2019, 10, 11023-1109.
(56) Pan, S.; Jiang, M.; Hu, J.; Xu, R.; Zeng, X.; Zhong, G. Syn hesis o
1,2-amino alcohols by deca boxyla i e coupling o amino acid de i ed α-amino
adicals o ca bonyl compounds ia isible-ligh pho oca alys in wa e . G een Chem.
2020, 22, 336-341.
(57) Guo, X.; Okamo o, Y.; Sch eie , M. R.; Wa d, T. R.; Wenge , O. S.
Enan ioselec i e syn hesis o amines by combining pho o edox and enzyma ic ca alysis
in a cyclic eac ion ne wo k. Chem. Sci. 2018, 9, 5052-5056.
(58) Von Lie , R. C. W.; de B ujin, A. D.; Roel es, G. A wa e -soluble
i idium pho oca alys o chemical modi ica ion o dehyd oalanine in pep ides and
p o eins. Chem. Eu . J. 2021, 27, 1430-1437.
(59) Nguyen, T.-T. H.; O’B ien, C. J.; Minh, L. N. T.; Olson, S. H.;
Se ine i, N. S.; P usine , S. B.; Pa as, N. A.; Con ad, J. Wa e soluble i idium
92
We hen p epa ed I (III)-con aining copolyme s a h ee i idium loading (LI )
egimes, P1−I 10, P1−I 23, and P1−I 40 (LI = 10, 23, and 40 mol % wi h espec o
AEMA uni s, espec i ely), by deco a ing he copolyme P1 h ough eac ion wi h he
C1 unde e y mild condi ions, a oom empe a u e and wi hou he use o any
addi i es (see Expe imen al Techniques o p ocedu e’s de ails).
Table 3.1. P ope ies o nea (P1) and unc ionalized copolyme s (P1-I x) syn hesized in his wo k
Sample
Mw
(kDa)a
Đb
AEMA
(mol%)c
LI
(mol%)d
P1
169.8
1.06
20
-
P1-I 10
178.6
1.10
18
10
P1-I 23
174.5
1.13
15.4
23
P1-I 40
211.7
1.03
12
40
a) Weigh -a e age molecula weigh . b) Dispe si y. c) Mola con en o AEMA uni s. d) I idium con en .
A second cyclome ala ed complex, bis[2-(2-py idinyl-N)phenyl-C](me hyl
ace oace a o)i idium(III) I (ppy)2(meacac) C2, was hen p epa ed ollowing a well-
known li e a u e p ocedu e,22 o be used as e e ence compound bo h o analy ic
pu poses and ca alysis pe o mance compa a ions, as i will be discussed la e . A e
de e mining he mola ex inc ion coe icien in chlo o o m o he complex C2 (Figu e
3.1), which was selec ed as model compound, i was possible o measu e by UV-Vis

93
spec opho ome y o polyme ic solu ions in chlo o o m ([polyme ] = 1 mg mL-1) he
quan i y o inco po a ed i idium(III) in he la e al chain o he polyme , by means o i s
con e sion in o he desi ed species shown in Table 1.
Figu e 3.1. On he le , supe imposed abso p ion spec a o he model compound
C2 in chlo o o m 6.5 10-5 < [C2] < 3.7 10-4 M in chlo o o m. On he igh , supe imposed
abso p ion spec a in chlo o o m o he h ee copolyme s P1-I x a polyme concen a ion o
1 mg mL-1, used o inco po a ed i idium LI quan i ica ion.
94
3.3.2. Sel -assembly in aqueous solu ion
Thanks o hei amphiphilic na u e, high molecula weigh (>150 kDa), and
inely uned composi ion, s able, sel -assembled SCNPs23,24 ha we deno ed as SCNP-
P1, APS-I 10, APS-I 23, and APS-I 40 we e ob ained upon simple dissolu ion o P1,
P1−I 10, P1−I 23, and P1−I 40, espec i ely, in wa e . Sel -assembly was asce ained by
measu ing ia dynamic ligh sca e ing (DLS) he di e ence ΔDh be ween
hyd odynamic diame e s Dh o P1, P1−I 10, P1−I 23, and P1−I 40 in e ahyd o u an,
THF, (good sol en o AEMA and OEGMA300) and hose o SCNP-P1, APS-I 10,
APS-I 23, and APS-I 40 measu ed in wa e (selec i e sol en o OEGMA300). i idium-
unc ionalized copolyme s all p esen ed a posi i e ΔDh (Table 3.2) because o he
o ma ion o a sel -collapsed a chi ec u e (see Figu e 3.2 and he Appendix: Sec ion
III).
Table 3.2. Hyd odynamic adii measu ed in THF (good sol en ) and wa e (selec i e sol en )
Sample
DhTHF (nm)
Sample
Dhwa e (nm)
P1
9.9
SCNP-P1
8.7
P1-I 10
12.3
APS-I 10
10.4
P1-I 23
11.8
APS-I 23
9.6
P1-I 40
10.4
APS-I 40
9.1
95
Figu e 3.2. Illus a ion o he educ ion in hyd odynamic size due o sel -assembly
o P1-I 23 (linea a chi ec u e) o APS-I 23 (SCNP a chi ec u e) on changing om THF (good
sol en ) o wa e (selec i e sol en ).
Addi ional e idence o he o ma ion o a sel -assembled con o ma ion in
APS-I 10, APS-I 23, and APS-I 40 was ob ained h ough pho oluminescence (PL)
expe imen s. As illus a ed in Figu e 3.3a, we measu ed he PL o eigh solu ions o he
model compound, bis(2-phenylpy idine)(me hyl ace oace ona e)i idium(III) C2 (see
he Appendix: Sec III), in THF / wa e mix u es upon inc ease o he wa e con en
( om 0% o 70%). We obse ed a dec ease in he PL in ensi y o C2 a high wa e
con en , which is consis en wi h li e a u e da a epo ed o he analogous complex
bis(2-phenylpy idine)- (ace ylace ona e)i idium(III), I (ppy)2(acac).25,26 Con e sely,
APS-I 10, APS-I 23, and APS-I 40 in wa e displayed signi ican PL in ensi y
enhancemen wi h espec o P1−I 10, P1−I 23, and P1−I 40 eco ded in wo di e en
nonselec i e sol en s (chlo o o m and THF) (see Figu e 3.3b).
96
Figu e 3.3. a) PL emission o model compound bis(2-phenylpy idine)(me hyl
ace oace ona e)i idium(III) C2 in THF / wa e mix u es ( om 0% o 70%) ([I (III)] = 3 μM,
λexc = 450 nm). b) Illus a ion o he agg ega ion-enhanced emission (AEE) o APS-I 40 in
wa e s P1–I 40 in CHCl3 and THF unde LED illumina ion ([I (III)] = 3 μM, λexc = 450 nm,
λem max = 521 nm).
These indings sugges ha ha con inemen o he hyd ophobic pho oca alys
in he limi ed space o he sel -assembled APS-I 10, APS-I 23, and APS-I 40 induces
signi ican agg ega ion-enhanced emission (AEE).
3.3.3. Pho oca aly ic ac i i y “in wa e ”
Wi h he sel -assembled SCNPs in hand, we es ed hei sui abili y as a i icial
pho osyn hases (APS) o in wa e pho oca alysis o a a ie y o o ganic
ans o ma ions, which we e selec ed om among he ple ho a o i idium(III)
cyclome ala ed complex-media ed eac ions in o ganic sol en s.27−29 Visible ligh has
97
been ecen ly applied success ully as an ene gy sou ce o [2 + 2] cycloaddi ions in
o ganic sol en s;30 he ein, we epo an unp eceden ed p ocedu e ha employs wa e
as he sole eac ion medium. Fo ins ance, an APS-I 40 solu ion was p epa ed by
dissol ing 2 mg o P1−I 40 in 1 mL o deionized wa e , which was hen cha ged wi h
58 μmol o he inylic compound 1a, and he esul ing mix u e was le s i ing a oom
empe a u e and unde LED illumina ion (λmax = 450 nm). A e his ime, he ex ac ed
c ude p oduc was analyzed ia quan i a i e 1H NMR (see he Appendix: Sec III) o
con e sion (c%) de e mina ion. Mul iple s a 3.54 and 3.98 ppm (see Figu e 3.4)
indica e he o ma ion o he 1,2-bis-subs i u ed cyclobu ane p oduc 2a, which was
hen isola ed in 90% yield as a mix u e o cis and ans dias e eome s.
Figu e 3.4. 1H NMR o he c ude o “in wa e ” [2+2] pho ocycloaddi ion eac ion
o 1a in he p esence o APS-I 40 (see ex ).
To ou deligh , we obse ed simila esul s when explo ing a a ie y o
subs a es, as illus a ed in Table 3.3. Conside ing ypical edox po en ials o inyl
a enes (e.g., Eox = 1.97 V s SCE, E ed = −2.53 V s SCE)31,32 and pho oca alys exci ed-
s a e po en ials (e.g., E*ox = 0.43 V s SCE and E* ed = −2.57 V s SCE),27 i seems
di icul o imagine he ac i a ion o ole ins 1a−1 by ypical single-elec on ans e
(SET).33 We hypo hesized ha , helped by he locally hyd ophobic packed en i onmen

98
o he APS, hese unlike p ocesses could be allowed ia an ene gy ans e (ET)
mechanism because o he simila iple ene gies o he in ol ed species (e.g., ET =
∼60 kcal mol−1 o inyl a enes, ET = ∼55 kcal mol−1 o C2).34
Table 3. Pho oca alyzed “in wa e ” [2+2] cycloaddi ion o inyl a enes (CA eac ion)a
En y
PC
T ans o ma ion
c%b
ans:cisc
1
APS-I 40
1a → 2a
96(90)d
1:0.3
2
APS-I 23
1a → 2a
94
1:0.3
3
APS-I 10
1a → 2a
96
1:0.3
4
APS-I 40
1b → 2b
97
1:0.2
5
APS-I 40
1c → 2c
96
1:0.3
6
APS-I 40
1d → 2d
90
1:0.3
7
APS-I 40
1e → 2e
60
1:0.3
8
APS-I 40
1 → 2
79
1:0.3
9
APS-I 40
1g → 2g
n.p.e
-
10
APS-I 40
1h → 2h
n.p e
-
11
No PC
1a → 2a
n.p.e
-
12
C2
1a → 2a
63
1:0.3
aSee Expe ime al Techniques o de ails. bCon e sion om 1H NMR. cDe e mined by 1H NMR.
dIsola ed yield in pa en hesis. eNo p oduc .
99
Byp oduc s o med in ace amoun s, among which we assigned35 he s uc u e
o 4-ace oxybenzaldehyde, ABA, o be he majo cons i uen in ag eemen wi h he
eac i i y o elec on- ich inyl a enes wi h eac i e oxygen species ha may be
gene a ed du ing i adia ion.36 Pe o mance o he same eac ion using he model
pho oca alys C2 d opped con e sion o 63% (see he Appendix o Chap e III) wi h he
emaining componen s o he c ude being he eac an and byp oduc s. No con e sion
was obse ed by employing wa e -soluble subs a es (1g, 1h, Table 3.3), p obably
because o di e ences in he ET alues.
In e es ingly, when 9-subs i u ed an h acenes we e employed as he subs a e,
we obse ed he o ma ion o an h aquinone 4. Since single oxygen and o he eac i e
oxygen species (ROS) a e gene a ed upon exci a ion o C2 in o ganic sol en s unde
ae obic condi ions,37 we su mised ha APS-I 10, APS-I 23, and APS-I 40 could be
e icien pho o-ca alys s o he “in wa e ” oxida ion o 9-subs i u ed an h acenes 3 o
an h aquinone 4 ia eac ion wi h ROS species38 (see Table 3.4). The o ma ion o 4
ha was con i med by 1H NMR, which is consis en wi h li e a u e da a,39 esul ed in
being comple ely neglec ed when he complex is no con ined wi hin he SCNP (see
he Appendix o Chap e III).
100
Table 3.4. Pho oca alyzed “in wa e ” oxida ion o 9-subs i u ed an h acenes (OA eac ion)a
En y
PC
Subs a e
c%b
1
APS-I 40
3a
62 (58)c
2
APS-I 23
3a
21
3
APS-I 10
3a
19
4
APS-I 40
3b
59
5
No PC
3a
n.p.d
6
C2
3a
n.p.d
aSee Expe imen al Techinques o de ails. bCon e sion om 1H NMR. cIsola ed yield. dNo p oduc .
Nex , we e alua ed he capabili y o APS-I 10, APS-I 23, and APS-I 40 o
ca alyze isible-ligh -induced α-a yla ion o a ylamines ha , since i s disco e y
epo ed by McMillan e al.,40,41 ha e ne e been epo ed in wa e . Table 3.5, en ies
1−3 show he p ocedu e in o ganic sol en s. To enable he pho oca aly ic cycle o occu
in wa e , we p epa ed APS-I 40 by dissol ing 2 mg o P1−I 40 in 1 mL o degassed
deionized wa e . We hen cha ged he APS aqueous solu ion wi h 29 μmol o 6 and
wi h a la ge excess o sodium ace a e, NaOAc, (84 equi .). The esul ing mix u e was
deoxygena ed by h ee consecu i e acuum / a gon back ill cycles, inally 87 μmol o
5 we e added and le s i ing a oom empe a u e and unde blue LED (λmax = 450 nm)
i adia ion o 12 h. Unde hese condi ions, p oduc 7 was ob ained by using APS-I 40
as pho oca alys in 57% con e sion and 48% pu i ied yield (Table 3.5, en y 5).
101
Table 3.5. Pho oca alyzed “in wa e ” α-a yla ion o a ylamines (AA eac ion)a
En y
PC
mol% PC
[NaOAc] Mb
Sol en b
c%c
1
C2
0.5
0.06
DMA
>95(95)d
2
P1-I 40
0.5
0.06
DMA
>95
3
C2
0.5
0.06
MeOH
8
4
APS-I 40
2
0.06
H2O
n.p.e
5
APS-I 40
2
2.4
H2O
57(48)d
6
APS-I 23
2
2.4
H2O
22
7
APS-I 10
2
2.4
H2O
21
8
C2
2
2.4
H2O
n.p.e
9
APS-I 40
2
2.4
H2O+
DMA
33
10
APS-I 40
2
sa .
H2O
24
11
No PC
-
2.4
H2O
n.p.e
aSee Expe imen al Techinques o de ails. bNaOAc = sodium ace a e; DMA = dime hylace amide;
MeOH = me hanol. cCon e sion o 7 om 1H NMR. dIsola ed yield o 7 in pa en hesis. eNo p oduc .
Sa u a ed in NaOAc.
Con e sion was ound o dec ease upon educ ion o he i idium loading in he
APS pho oca alys (Table 3.5, en ies 6 and 7). We a ibu ed he s iking con e sion
d op upon ei he lowe ing o inc easing he base concen a ion (Table 3.5, en ies 4 and
10) o he al e ed acidi ies o he eac an s and addi i es in aqueous media wi h espec
o he eac ion condi ions epo ed in li e a u e o o ganic sol en .40,42
108
Figu e 3.9. Michaelis-Men en plo o he “in wa e ” [2+2] pho ocycloaddi ion o
1a ca alyzed by APS-I 40.
An appa en ca aly ic cons an kca ,app o 2.6 s-1 was ob ained om Equa ion 3.3:
𝑘𝑐𝑎𝑡,𝑎𝑝𝑝(𝑠−1)=𝑉
𝑚𝑎𝑥
𝑁𝑇
Equa ion 3.3
whe e Vmax is he maximum measu ed eloci y o con e sion o 1a in o he i le
p oduc 2a and NT is he nanopa icle concen a ion employed o he ca aly ic eac ion,
calcula ed aking he Mw (SEC) as molecula weigh . The alue o he appa en
Michaelis-Men en cons an ob ained was KM,app = 4.6 × 10-2 M.
3.3.6. Recyclabili y o APS
To assess he eusabili y o APSs, we pe o med a ecyclabili y expe imen
agains he “in wa e ” [2+2] pho ocycloaddi ion o inyl a enes in he bes condi ions
in e ms o p oduc yields. A e a 1s “in wa e ” [2+2] pho ocycloaddi ion eac ion o

109
1a wi h APS-I 40 as ca alys , he o ganic p oduc s we e ex ac ed by h ee consecu i e
imes wi h 3 mL o die hyl e he each. The collec ed o ganic ac ions we e d ied o e
anhyd ous magnesium sul a e, il e ed, and concen a ed unde educed p essu e. The
ex ac ed c udes we e hen di ec ly dilu ed wi h 0.5 mL o CDCl3 o quan i a i e NMR
analysis. The aqueous phase was hen collec ed and cen i uged a 4°C, he pale-yellow
sedimen was e-dispe sed in 1 mL o deionized wa e by means o con inuous s i ing
a . . and in he da k o 1 h. Fo he 2nd ca aly ic cycle, he esul ing clea , aded-yellow
solu ion was subsequen ly ans e ed o a 4 mL eac ion essel, cha ged wi h 58 µmol
o inyl a ene 1a and le open and s i ing a . . and unde LED illumina ion (λmax =
450 nm) o 12 hou s. A e his ime, he o ganic p oduc s we e ex ac ed by h ee
consecu i e imes wi h 3 mL o die hyl e he each. The collec ed o ganic ac ions we e
d ied o e anhyd ous magnesium sul a e, il e ed, and concen a ed unde educed
p essu e. The ex ac ed c udes we e hen di ec ly dilu ed wi h 0.5 mL o CDCl3 o
quan i a i e NMR analysis. Nex 3 d and 4 h ca aly ic cycles we e ca ied ou ollowing
he same p ocedu e as desc ibed o he 2nd one. Figu e 3.10 illus a es he esul s o
he ecyclabili y o APS-I 40 in 4 consecu i e cycles.
Figu e 3.10. Resul s o he ecyclabili y expe imen o APS-I 40 as ca alys o he
“in wa e ” [2+2] pho ocycloaddi ion o 1a.
0
20
40
60
80
100
1 2 3 4
2a : 1a (mol/mol)
Nº cycle
110
3.4. Expe imen al Techniques
3.4.1. Sol en s and eagen s
Unless o he wise no ed, all eagen s and sol en s we e used as ecei ed om
endo s. (Oligoe hylene glycol monome hyl e he ) me hac yla e (OEGMA300, a e age
molecula weigh = 300 Da) (>99%) and 4-ace oace oxye hyl me hac yla e (AEMA)
(>95%) we e pu chased om TCI Eu ope N.V. and we e il e ed o e basic alumina
be o e use. n-Hexane (96%), ace one (>99%), me hylene chlo ide (CH2Cl2) (>99%,
+0.2 % E OH), n-pen ane (99%), e hyl ace a e (AcOE ) (>99.8) and die hyl e he
(E 2O) (>99%, +7 ppm BHT) we e pu chased om Scha lab. Te ahyd o u an (THF)
(>99 %, +0.025% BHT) and me hanol (MeOH) (>99%) we e pu chased om Fishe
Scien i ic, inhibi o - ee THF was ob ained by il a ion o e basic alumina. 1,4-
Dioxane (>99%), me hyl ace oace a e (99%), 2-e hoxye hanol (≥99%), sodium
hyd oxide (NaOH) (≥98%), chlo o o m (>99%, +100-200 ppm amylene), α-me hyl
s y ene (99%), 2-naph alenesul onyl luo ide (95%), sodium ace a e (NaOAc) (>99%),
anhyd ous magnesium sul a e (MgSO4) (>99%), sodium chlo ide (NaCl) (>99%),
sil e (I) i luo ome hansul ona e (AgOT ) (≥98%), 4-cyano-4-
( hiobenzoyl hio)pen anoic acid (CPADB), 1,4-dicyanobenzene (DCB) (98%),
ie hylamine (TEA) (>99%), linalool (97%) and dime hylace amide (DMA) (≥99%)
we e pu chased om Sigma-Ald ich. p-Toluenesul onyl chlo ide (99%) and p-
oluenesul onyl b omide (98%) we e pu chased om Sigma-Ald ich and handled
unde a gon a mosphe e. 4-Ace oxys y ene (96%), 4-chlo os y ene (99%), 3-
b omos y ene (97%), 4- i luo ome hyls y ene (99%), 4- inylpy idine (≥95%) and 1-
inyl-1,2,4- iazole (≥97%) we e pu chased om Sigma-Ald ich and used a e
il e ing o e basic alumina. Bis(μ-chlo o) e akis(2- phenyl-py idina o)dii idium(III)
(98%), i idium(III) chlo ide ihyd a e (98%), 2-phenylpy idine (>99%), 4-
ca boxys y ene (>99%) and 4-me hoxys y ene (>99%) we e pu chased om BLD
Pha ma. Azobisisobu y oni ile (AIBN) (98%) was pu chased om Fluka and
ec ys allized om MeOH p io use. Silica gel o column ch oma og aphy (0.035-0.07
nm 60 A) was pu chased om Ac os O ganics. Basic alumina (0.063-0.2 mm) was
pu chased om Me ck. N-phenylpy olidine (>98%) was pu chased om Alpha Aesa .
Deu e a ed chlo o o m (CDCl3, 99.8% D, + 0.03% e ame hylsilane) o NMR
111
analysis was pu chased om Eu iso op. Deionized wa e was ob ained om a
The moscien i ic Ba ns ead TII Sys em.
3.4.2. Analy ical me hods and echniques
Nuclea Magne ic Resonance (NMR) Spec oscopy: 1H and 13C NMR spec a
we e eco ded a oom empe a u e ( . .) on a B uke spec ome e ope a ing a 400
MHz, using CDCl3 as sol en .
- Hyd ophobic monome con en in he copolyme , %AEMA (mol%), was
calcula ed acco ding o Equa ion 3.4:
%𝐴𝐸𝑀𝐴 (𝑚𝑜𝑙%)= 𝑆𝐴𝐸𝑀𝐴
𝑂𝑀𝑒
𝑆𝑂𝐸𝐺𝑀𝐴
𝑂𝑀𝑒 + 𝑆𝐴𝐸𝑀𝐴
𝑂𝑀𝑒 ×100
Equa ion 3.4
whe e SOMeAEMA is he in eg a ed a ea o he signal co esponding o he
me hoxylic p o ons o he AEMA uni s wi hin he copolyme (2.29 ppm) and
SOMeOEGMA is he in eg a ed a ea o he signal co esponding o he me hoxylic p o ons
o OEGMA uni s (3.37 ppm).
- NMR con e sion o he pho o [2+2] cycloaddi ion eac ion, c% (mol%), was
calcula ed acco ding o Equa ion 3.5:
𝑐% (𝑚𝑜𝑙%)= 𝛴𝑃
𝑆𝑅+ 𝛴𝑃×100
Equa ion 3.5
whe e ΣP is he sum o he no malized in eg a ed a ea signals o he alkylic
p o ons o all he isome s o he p oduc (3.56 ppm and 4.02 ppm) and SR is he
no malized a ea o he signal o he inylic p o on o he eac an (5.74-5.69 ppm). The
dias e eome ic a io d. . was calcula ed using he in eg a ed a ylic p o on signals o he
p oduc (7.03-7.01 ppm o he ans isome and 6.86-6.83 ppm o he cis isome ).
- NMR con e sion o he α-a yla ion o a ylamines, c% (mol%), was calcula ed
acco ding o Equa ion 3.6 o Equa ion 3.7 when an in e nal s anda d was used:
112
𝑐% (𝑚𝑜𝑙%)= 𝑆𝑃
𝑆𝐷𝐶𝐵
𝐴𝑟 + 𝑆𝑃×100
Equa ion 3.6
𝑐% (𝑚𝑜𝑙%)= 𝑆𝑃𝑛𝑆𝐼
𝑆𝑆𝐼 + 𝑛0
𝑃×100
Equa ion 3.7
whe e SP is he no malized a ea o he a ylic signals o he p oduc (7.60-7.58
ppm), SA DCB is he no malized a ea o he a oma ic signal o he eac an 1,4-
dicyanobenzene (7.79 ppm), nSI is he in e nal s anda d (linalool) numbe o moles
co esponding o he weigh ed amoun in he ube, SSI is he no malized in ensi y o he
signal o he in e nal s anda d (5.95-5.89 ppm) and nP0 is he numbe o moles o he
eac an 1,4-dicyanobenzene co esponding o he weigh ed amoun be o e he
eac ion.
- NMR con e sion o he oxida ion o 9-subs i u ed an h acenes, c% (mol%),
was calcula ed acco ding o Equa ion 3.8:
𝑐% (𝑚𝑜𝑙%)= 𝑆𝑃
𝑆𝑅+ 𝑆𝑃×100
Equa ion 3.8
whe e SP is he no malized a ea o he a ylic signals o he an h aquinone
p oduc (7.81-7.79 ppm), SR is he no malized a ea o he signal o he eac an (e.g,.
signal a 8.04-8.01 ppm o he 9-hyd oxyme hyl an h acene).
- NMR con e sion o he β-hyd oxysul onyla ion eac ion, c% (mol%), was
calcula ed acco ding o Equa ion 3.9:
𝑐% (𝑚𝑜𝑙%)= 𝑆𝑃𝑛𝑆𝐼
𝑆𝑆𝐼 + 𝑛0
𝑃×100
Equa ion 3.9
whe e SP is he no malized a ea o he a ylic signals o he p oduc (7.49-7.48
ppm), nSI is he in e nal s anda d (1,4-dicyanobenzene) numbe o moles co esponding
o he weigh ed amoun in he ube, SSI is he no malized in ensi y o he signal o he
in e nal s anda d (7.79 ppm) and nP0 is he numbe o moles o he eac an α-me hyl
s y ene co esponding o he amoun be o e he eac ion.
113
Size-Exclusion Ch oma og aphy (SEC): SEC measu emen s we e pe o med a
30 °C on an Agilen 1200 sys em equipped wi h PLgel 5m Gua d and PLgel 5m
MIXED-C columns, and iple de ec ion: a di e en ial e ac i e index (dRI) de ec o
(Op ilab Rex, Wya ), a mul i-angle lase ligh sca e ing (MALLS) de ec o
(MiniDawn T eos, Wya ), and a iscosime ic (VIS) de ec o (ViscoS a -II, Wya ).
Da a analysis was pe o med wi h ASTRA So wa e ( e sion 6.1) p o ided by Wya .
THF was used as eluen a a low a e o 1 mL min-1. A alue o dn/dc = 0.115 mLg-1
was used o copolyme P1 and de i a i es he eo .
Dynamic Ligh Sca e ing (DLS): DLS measu emen s we e ca ied ou a . .
on a Mal e n Ze asize Nano ZS appa a us, using high p ecision qua z cells, ligh pa h
10×10 mm, p o ided by Hellma Analy ics. Da a a e gi en as by Numbe as an a e age
o a leas ou measu emen s. All he sol en s o sample p epa a ion we e il e ed wi h
2 µM Te lon il e s p io use.
UV-Vis Spec oscopy (UV-Vis): UV-Vis spec a we e eco ded a 25 °C in an
Agilen 8453A appa a us wi h Pel ie he mos a ic cell holde , T-con olle 89090A,
using high p ecision qua z cells, ligh pa h 10×10 mm, p o ided by Hellma Analy ics.
Pho oluminescence (PL) Spec oscopy: PL spec a we e eco ded a . . on an
Agilen Ca y Eclipse spec ome e a an exci a ion wa eleng h o 365 nm, using high
p ecision qua z cells, ligh pa h 10×10 mm, p o ided by Hellma Analy ics. Samples
we e degassed by pu ging a gon gas o 5 consecu i e minu es be o e each
measu emen .
Pho oca aly ic Reac ions, I adia ion Se -Up: Pho o eac ions we e ca ied ou
using a Penn PhD Pho o eac o , equipped wi h a 450 nm LED sou ce, pu chased om
Me ck. LED in ensi y was se a 100%, and a 4 mL ial-holde was used (10 cm ligh
pa h o he eac ion essel and o m he ligh sou ce).

114
3.4.3. Syn hesis and cha ac e iza ion o compounds
3.4.3.1. Syn hesis o he Polyme ic P ecu so Poly(OEGMA300-co-AEMA) (P1)
In an o en d ied Schlenk lask equipped o a magne ic s i ba , 1.92 g (6.4
mmol) o OEGMA300, 343 mg (1.6 mmol) o AEMA, 10.6 mg (38 µmol) o 4-cyano-
4-( hiobenzoyl hio)pen anoic acid (CPADB), 1.29 mg (7.7 µmol) o AIBN and 3.4 mL
o 1,4-dioxane we e added in his o de . The lask was hen sealed wi h a ubbe sep um
and, a e pu ging he solu ion wi h a gon low o 20 min., he eac ion mix u e was
le s i ing a 70ºC unde a gon a mosphe e o 19 h. A e his ime, he eac ion was
quenched subme ging he ube in liquid ni ogen. The c ude was hen p ecipi a ed in a
la ge excess o n-hexane o h ee consecu i e imes, yielding he desi ed polyme ic
p oduc P1. Mw (kDa) = 169.8, PDI = 1.06, %AEMA (mol%) = 20, 1H NMR (400 MHz,
CDCl3): δ (ppm) = 4.33 (m., CH3COCH2CO), 4.14 (m., CH2CO2CH2CH2), 4.07 (m.,
CH2CO2C), 3.65-3.54 (m., OCH2CH2O), 3.37 (COCH3), 2.29 (s., CH3COCH2), 2.09-
1.17 (m., CH2CCH3), 1.01-0.86 (m., CH2CCH3). 13C NMR (100 MHz, CDCl3): δ (ppm)
= 30.42, 44.90, 45.26, 49.75, 59.12, 64.03, 68.58, 70.69, 72.05.
115
3.4.3.2. Syn hesis o he Hyd oxo-B idged I idium(III) Dime Te akis(2-
phenylpy idina o-N,C2)(m-dihyd oxy)dii idium(III) ([I (ppy)2OH]2) (C1)
The dime ic complex C1 was syn hesized acco ding o a p ocedu e epo ed in
li e a u e.21 A mix u e o 257 mg (0.73 mmol) o I Cl3 3H2O, 276 mg (1.71 mmol) o
2-phenyl-py idine in 12 ml o a solu ion o 2-e hoxy-e hanol/wa e (3:1) was e luxed
unde a gon o 4.5 h. A e his ime, an excess o NaOH (1.25 g, 30 mmol) dissol ed
in 12.5 mL o H2O was added and he esul ing mix u e was le s i ing unde e lux
o 2 h. A e cooling o . ., 25 mL o H2O was added. An o ange-b own p ecipi a e
was il e ed o , dissol ed in 15 mL me hylene chlo ide and, subsequen ly, il e ed. The
il a e was ea ed wi h a NaOH solu ion (1.68 g, 0.04 mol in 4.5 mL H2O) a e lux
o 6 h. A e wa ds, he o ganic sol en (CH2Cl2) was e apo a ed, and H2O (125 mL)
was added. The c ude p oduc was il e ed o and washed wi h n-pen ane (10 mL) and
die hyl e he (10 mL). Fu he pu i ica ion was ca ied ou by p ecipi a ion (CH2Cl2
solu ion) in n-pen ane, yielding [I (ppy)2OH]2 (C1) as a b own powde (295 mg, 78%).
1H NMR (400 MHz, CDCl3): δ (ppm) = 9.4-9.23 (m, 1H, Hk), 8.69-8.47 (m, 1H, Hh),
7.9-7.45 (m, 2H, He,i), 6.84-6.51 (m, 3H, Hc,d,j), 6.02-5.83 (m, 1H, Hb), –1.54 (s, 1H,
Hl).
116
3.4.3.3. Syn hesis o he Cyclome ala ed Complex Bis[2-(2-py idinyl-
N)phenyl-C](me hyl ace oace a o)i idium(III) (C2)
The complex C2 was syn hesized as ollows and acco ding o a p ocedu e o
he p epa a ion o he analogue bis[2-(2-py idinyl-N)phenyl-
C](ace ylace ona o)i idium(III), which is well desc ibed in li e a u e.22 Speci ically,
117.9 mg (0.11 mmol) o [I (ppy)2Cl]2 and 84 mg (0.32 mmol) AgOT we e dissol ed,
in his o de , in 8 mL o degassed ace one and e luxed a 55ºC unde ni ogen
a mosphe e and con inuous s i ing o 2 h. The solu ion was cooled o . . and il e ed
o emo e AgCl. The il a e was e luxed unde ni ogen a mosphe e o 1 h and added
unde ine a mosphe e o a 1 h e luxed solu ion o me hyl ace oace a e (46 µl, 0.43
mmol) and ie hylamine (113 µl, 0.81 mmol) dissol ed in degassed ace one (4 ml),
The esul ing b igh -b own solu ion was e luxed o e nigh unde ni ogen a mosphe e.
The c ude was hen cooled o . . and il e ed on co on o elimina e las esidues o
AgCl. The ola iles we e emo ed and he solid was sonica ed in deionized wa e and
cen i uged o h ee consecu i e imes. The ob ained solid was hen d ied unde
acuum a . . o h ee days, a o ding he desi ed p oduc C2. Yield % (weigh %): 85
%. 1H NMR (400 MHz, CDCl3): δ (ppm) = 8.64-8.53 (m, 2H, Hk), 7.90-7.84 (m, 2H,
Hh), 7.79-7.73 (m, 2H, Hi), 7.58-7.53 (m, 2H, He), 7.21-7.15 (m, 2H, Hj), 6.86-6.80 (m,
2H, Hd), 6.73-6.86 (m, 2H, Hc), 6.29-6.25 (m, 2H, Hb), 4.72 (s, 1H, Hm), 3.40 (s, 3H,
Hl), 1.83 (s, 3H, Hn). 13C NMR (100 MHz, CDCl3): δ (ppm) = 28.73, 46.67, 51.28,
83.19, 99.95, 118.10, 118.39, 119.18, 120.60, 120.69, 121.42, 121.52, 123.49, 123.87,
128.78, 132.92, 133.10, 136.75, 136.96, 148.62, 148.80, 168.73, 186.37.
117
3.4.3.4. Syn hesis o I idium(III)-Deco a ed Copolyme s a Di e en
I idium(III) Loadings (P1-I 40, P1-I 23 and P1-I 10)
50 mg (0.707 mmol o AEMA) o polyme p ecu so P1 and 18.6 mg (17.7
µmol) o C1 we e dissol ed in 2 mL o anhyd ous chlo o o m. The mix u e was hen
degassed o 10 minu es and le s i ing a . . unde ine a mosphe e and in he da k
o 2 days. A e his ime, he c ude was il e ed and he ola iles we e emo ed luxing
ni ogen, un il a deep yellowish-b own ilm was ob ained. P1-I 40: I idium(I )(III)
loading, LI (mol%) = 40, Mw (kDa) = 211.7, PDI = 1.03. P1-I 23 was ob ained ollowing
he same p ocedu e epo ed abo e, employing 50 mg o polyme p ecu so P1 and 9.3
mg (8.8 µmol) o C1. P1-I 23: LI (mol%) = 23, Mw (kDa) = 174.5, PDI = 1.13. P1-I 10
was ob ained ollowing he same p ocedu e epo ed abo e, employing 50 mg o
polyme p ecu so P1 and 4 mg (3.8 µmol) o C1. LI (mol%) = 10, Mw (kDa) = 178.6,
PDI = 1.10. 1H NMR (400 MHz, CDCl3) (P1-I 40): δ (ppm) = 8.55-8.50 (m, 1 H), 7.84-
7.75 (m, 1 H), 7.50 (m, 2H), 7.20 (m, 1 H), 6.77 (m, 1 H), 6.63 (m, 1 H), 6.19 (m, 1 H),
4.73 (m, 1 H), 4.31 (m., CH3COCH2CO), 4.13 (m., CH2CO2CH2CH2), 4.05 (m.,
CH2CO2C), 3.62-3.51 (m., OCH2CH2O), 3.34 (COCH3), 2.27 (s., CH3COCH2), 2.09-
1.78 (m., CH2CCH3), 0.99-0.85 (m., CH2CCH3). 13C NMR (100 MHz, CDCl3): δ (ppm)
= 30.79, 45.46, 45.76, 50.30, 59.69, 64.60, 69.14, 71.25, 72.60.
124
NaOAc espec i ely. AA-9 was ca ied ou ollowing he gene al p ocedu e, using 1
mL o bina y solu ion wa e /DMA (DMA 10%) as sol en .
α-A yla ion o N-A ylamines in Wa e . Non-suppo ed ca alys p ocedu e (AA-
8). To a 4 mL o en d ied ial, equipped o a magne ic s i ba , was added 0.36 mg (0.58
µmol) o C2, 3.7 mg (29 µmol) o 1,4-dicyanobenzene and 203 mg (2.47 mmol) o
sodium ace a e. The ial was hen sealed wi h a ubbe sep um. A e h ee consecu i e
acuum-e acua ion/A gon back ill cycles, 1 mL o ex ensi ely degassed deionized
wa e and 12.5 µL (87 µmol) o N-phenyl py olidine we e added unde A gon posi i e
p essu e. The esul ing mix u e was le s i ing a . . and unde LED illumina ion (λmax
= 450 nm) o 12 hou s. A e his ime, he mix u e was ex ac ed h ee imes wi h 6
mL o e hyl ace a e each. The collec ed o ganic ac ions we e d ied o e anhyd ous
magnesium sul a e, il e ed, and concen a ed unde educed p essu e. The ex ac ed
c ude was hen dilu ed wi h 6.5 mL o CDCl3 o quan i a i e NMR analysis. No
con e sion o he desi ed p oduc was obse ed.
α-A yla ion o N-A ylamines in Wa e . P ocedu e in o ganic sol en s (AA-1,
AA-2, AA-3). An o en-d ied 15 mL ial equipped wi h a ubbe sep um and a magne ic
s i ba was cha ged wi h 0.5% mol o pho oca alys (3.08 mg o C2 o AA-1 and AA-
3, 50 µL o 20 mgmL-1 P1-I 40 s ock solu ion in me hylene chlo ide o AA-2), 128.1
mg (1 mmol) o 1,4-dicyanobenzene and 164,1 mg (2 mmol) o NaOAc. The Schlenk
was hen pu ged wi h 3 consecu i e acuum e acua ion/a gon back ill. Then, 4 mL o
degassed sol en (DMA o AA-1, AA-2, MeOH o AA-3) was added unde posi i e
ni ogen p essu e ollowed by 434 µL (3 mmol) o N-phenylpi olidine. The eac ion
mix u e was u he ly degassed ia h ee cycles o acuum e acua ion/a gon back ill.
A e degassing, he ial was sealed wi h pa a ilm and le unde LED illumina ion
(λmax = 450 nm) o 12 hou s. A e his ime, he eac ion was hen dilu ed wi h e hyl
ace a e and added o a sepa a o y unnel con aining 25 mL o a sa u a ed Na2CO3
aqueous solu ion. The laye s we e sepa a ed, and he aqueous laye was ex ac ed wi h
E OAc. The collec ed o ganic ac ions we e d ied o e anhyd ous magnesium sul a e,
il e ed, and concen a ed unde educed p essu e. The ex ac ed c ude was hen

125
pu i ied ia silica gel column ch oma og aphy o a o d he i le compound 7 (n-
hexane/e hyl ace a e 10:1 o 2:1).
126
3.4.7. “In wa e ” β-Hyd oxysul onyla ion o α-Me hyl S y ene
Gene al P ocedu e
200 µL o a s ock solu ion o polyme ic ca ie P1-I 23 in me hylene chlo ide
([polyme ] = 20 mg/mL) and 9 mg (35 µmol) o osyl chlo ide we e pu in an o en
d ied 4 mL ial, p e iously equipped wi h a magne ic s i ba . A e comple e emo al
o he ola iles, he ial was sealed wi h a ubbe sep um and he esul ing polyme ic
ilm was degassed ia h ee consecu i e acuum-pump/A gon-back ill cycles. 1 mL o
ex ensi ely degassed deionized wa e and 3.4 mg (29 µmol) o α-me hyl s y ene we e
hen added unde A gon posi i e p essu e. The esul ing solu ion was le s i ing a . .
and unde LED ligh (λmax = 450 nm) i adia ion o 12 hou s. A e his ime, he c ude
was ex ac ed h ee imes wi h 6 mL o e hyl ace a e each. The collec ed o ganic
ac ions we e d ied o e anhyd ous magnesium sul a e, il e ed, and concen a ed
unde educed p essu e. A e sol en emo al, he c ude p oduc was ei he pu i ied
by column ch oma og aphy on silica gel (n-hexane/e hyl ace a e 4:1) o a o d he i le
compound 10a o added o a known amoun o 1,4-dicyanobenzene and dilu ed wi h
0.6 mL o CDCl3 o quan i a i e 1HNMR analysis. Isola ed yield (weigh %) = 58 %.
Con e sion NMR (mol%) = 67 %. 1H NMR (400 MHz, CDCl3): δ (ppm) = 7.49 (d,
2H), 7.29 – 7.27 (m, 2H), 7.18 ( d, 5H), 4.63 (b .s., 1H), 3.71 – 3.57 (m, 2H), 2.39 (s,
3H), 1.71 (s, 3H). 13C NMR (100 MHz, CDCl3): δ (ppm) = 21.73, 29.85, 60.54, 62.31,
121.88, 126.38, 128.08, 128.51, 128.84, 129.65, 136.76, 142.43.
HS-1 was ca ied ou acco ding o he gene al p ocedu e, using 100 µL o P1-
I 40 s ock solu ion in me hylene chlo ide ([polyme ] = 20 mg/mL). HS-3 was ca ied
127
ou acco ding o he gene al p ocedu e, using 400 µL o P1-I 10 s ock solu ion in
me hylene chlo ide ([polyme ] = 20 mg/mL). HS-4 was ca ied ou ollowing he
gene al p ocedu e using 8.2 mg (35 µmol) o pa a oluensul onyl b omide in
subs i u ion o he osyl chlo ide. HS-5 was ca ied ou ollowing he gene al p ocedu e
using 7.4 mg (35 µmol) o 2-naph alenesul onyl luo ide in subs i u ion o osyl
chlo ide.
β-Hyd oxysul onyla ion o A oma ic Alkenes. Non-suppo ed ca alys
p ocedu e (HS-6). To a 4 mL o en d ied ial, equipped o a magne ic s i ba , was
added 0.36 mg (0.58 µmol) o C2, 9 mg (35 µmol) o osyl chlo ide we e pu in an o en
d ied 4 mL ial, p e iously equipped wi h a magne ic s i ba . A e comple e emo al
o he ola iles, he ial was sealed wi h a ubbe sep um and he esul ing polyme ic
ilm was degassed ia h ee consecu i e acuum-pump/A gon-back ill cycles. 1 mL o
ex ensi ely degassed deionized wa e and 3.4 mg (29 µmol) o α-me hyl s y ene we e
hen added unde A gon posi i e p essu e. The esul ing solu ion was le s i ing a . .
and unde LED ligh (λmax = 450 nm) i adia ion o 12 hou s. A e his ime, he c ude
was ex ac ed h ee imes wi h 6 mL o e hyl ace a e each. The collec ed o ganic
ac ions we e d ied o e anhyd ous magnesium sul a e, il e ed, and concen a ed
unde educed p essu e. A e sol en emo al, he c ude p oduc was added o a known
amoun o 1,4-dicyanobenzene and dilu ed wi h 0.6 mL o CDCl3 o quan i a i e
1HNMR analysis.
HS-7 was ca ied ou ollowing he gene al p ocedu e and wi hou adding any
pho oca alys . No con e sion o he desi ed p oduc was obse ed.
128
3.5. Conclusions
In summa y, a i icial pho osyn hases (APS) a ise by endowing enzyme-
mime ic single-chain nanopa icles, SCNPs, wi h b oad isible-ligh pho oca aly ic
ac i i y o challenging “in wa e ” o ganic eac ions. We in oduce a i s gene a ion o
APS esul ing om he deco a ion o an amphiphilic high-molecula -weigh
copolyme , poly- (OEGMA300- -AEMA), wi h i idium(III) cyclome ala ed complex
pendan s ollowed by i s own sel -assembly in wa e . APS enabled e icien isible-
ligh pho oca alysis o a a ie y o o ganic ans o ma ions in an aqueous solu ion a
oom empe a u e and unde LED illumina ion (λmax = 450 nm). This wo k b oadens
he possibili ies o pe o ming challenging “in wa e ” o ganic ans o ma ions ia
APS-media ed isible-ligh pho oca alysis.
129
3.6. Re e ences
(1) Emmanuel, M. A.; Bende , S. G.; Bilodeau, C.; Ca celle , J. M.; DeHo i z,
J. S.; Fu H.; Liu, Y.; Nicholls B. T.; Ouyang, Y.; Page, C. G.; Qiao, T.; Raps, F. C.;
So igué, D. R.,; Sun, S.-Z.; Tu ek-He man, J.; Ye, Y.; Ri as-Suche , A.; Cao, J.;
Hys e , T. K. Pho obioca aly ic s a egies o o ganic syn hesis. Chem. Re . 2023, 123,
5459-5520.
(2) Alphand, V.; Van Be kel, W. J. H.; Ju kaš, V.; Ka a, S.; Kou is , R.;
K ou il, W.; Mascia, F.; Nowaczyk, M. M.; Paul, C. E.; Schmid , S.; Spasic, J.;
Tamagnini, P.; Winkle , C. K. Exci ing enzymes: Cu en s a e and u u e pe spec i e
o pho obioca alysis. ChemPho oChem 2023, 7, e202200325.
(3) Sanca , A. S uc u e and unc ion o DNA pho olyase. Biochemis y 1994,
33, 2-9.
(4) T imble, S. J; C awshaw, R; Ha dy, F. J.; Le y, C. W.; B own, M. J. B;
Fue s , D. E.; Heyes, D. J.; Obexe , R.; G een, A. P. A designed pho oenzyme o
enan ioselec i e [2+2] cycloaddi ions. Na u e 2022, 611, 709-714.
(5) Jeong, W. J.; Lee, J.; Eom, H.; Song, W. J. A speci ic guide o
me alloenzyme designe s: in oduc ion and e olu ion o me al-coo dina ion sphe es
embedded in p o ein en i onmen s. Acc. Chem. Res. 2023, 56, 2416-2425.
(6) Fu, Y.; Huang, J.; Wu, Y.; Liu, X.; Zhong, F.; Wang, J. Bioca aly ic c oss-
coupling o a yl halides wi h a gene ically enginee ed pho osensi ize a i icial
dehalogenase. J. Am. Chem. Soc. 2021, 143, 617-622.
(7) Russo, C.; B unelli, F.; T on, C. G.; Gius iniano, M. Visible-ligh
pho o edox ca alysis in wa e . J. O g. Chem. 2023, 88, 6284-6293.
(8) Sun, K.; L , Q.-Y.; Chen, X.-L.; Qu, L.-B., Yu, B. Recen ad ances in
isible-ligh -media ed o ganic ans o ma ions in wa e . G een Chem. 2021, 23, 232-
248.
(9) Hong, D.; Shi, L.; Liu, X.; Ya, H.; Han, X. Pho oca alysis in wa e -soluble
sup amolecula me al o ganic complex. Molecules 2023, 28, 4068.
(10) Bo ecchia, C.; Noël, T. Pho oca aly ic modi ica ion o amino acids,
pep ides and p o eins. Chem. Eu . J. 2019, 25, 26-42.
(11) Bu, M.-J.; Cai, C.; Gallou, F.; Lipshu z, B. H. PQS-enabled isible-ligh
i idium pho o edox ca alysis in wa e a oom empe a u e. G een Chem. 2018, 20,
1233-1237.

130
(12) Mundsinge K.; Tu en B. T.; Wang, L.; Neubane , K.; K op , C.; O’Ma a,
M. L.; Ba ne -Kowollik, C. Visible-ligh - eac i e single-chain nanopa icles. Angew.
Chem. In . Ed. 2023, 62, e202302995.
(13) Pomposo, J. Single-chain polyme nanopa icles: syn hesis,
cha ac e iza ion, and applica ions. Wiley-VSH 2017, 978-3-527-80638-6.
(14) Ro h uss, H.; Knö el, N. D.; Roesky, P. W.; Ba ne -Kowollik, C. Single-
chain nanopa icles as ca aly ic nano eac o s. J. Am. Chem. Soc. 2018, 140, 5875-5881.
(15) Rubio-Ce illa, J.; González, E.; Pomposo, J. A. Ad ances in single chain
nanopa icles o ca alysis applica ions. Nanoma e ials 2017, 7, 341.
(16) Ve de-Ses o, E.; A be, A.; Mo eno, A. J.; Cangialosi, D.; Aleg ía, A.;
Colmene o, J.; Pomposo, J. A. Single-chain nanopa icles: oopo uni ies p o ided by
in e nal and ex e nal con inemen . Ma e . Ho iz. 2020, 7, 2292-2313.
(17) Xiong, T. M.; Ga cia, E. S.; Chen, J.; Zhu, L.; Alzona, A. J.; Zimme mann,
S. C. Enzyme-like ca alysis by single chain nanopa icles ha use ansi ion me al
co ac o s Chem. Commun. 2022, 58, 985-988.
(18) Mundsinge , K.; Izuagbe, A.; Tu en, B. T.; Roesky, P. W.; Ba ne -
Kowollik, C. Single chain nanopa icles in ca alysis. Angew. Chem. In . Ed. 2023,
e202311734.
(19) Chen, R.; Be da, E. 100 h anny e sa y o mac omolecula science
iewpoin : e-examining single-chain nanopa icles. ACS Mac o Le . 2020, 9, 1836-
1843.
(20) Spicuzza, M.; Gaikwad, S. P.; Huss, S.; Lee, A.; C aescu, C. V.; G iggs,
A.; Joseph, J.; Pu henpu ayil, M.; Lin, W.; Ma a azzo, C.; Baldwin, S.; Pe ez, V.;
Rod iguez-Ace edo, D. A.; Swie k, J. R.; Elaqua E. Visible-Ligh -Media ed Diels-
Alde Reac ions Unde Single-Chain Polyme Con inemen : In es iga ing he Role o
he C osslinking Moie y on Ca alys Ac i i y. ChemRxi , 2023, 10.26434/chem xi -
2023-zw sq- 2.
(21) Ulb ich , C.; Bece , C. R.; Win e , A.; Schube , U. S. RAFT
polyme iza ion mee s coo dina ion chemis y: syn hesis o a polyme -based
i idium(III) emi e . Mac omol. Rapid Commun. 2010, 31, 827-833.
(22) Beye , B.; Ulb ich , C.; Win e , A.; Hage , M. D.; Hoogenboom, R.;
He ze , N.; Baumann, S. O.; Kickelbick, G.; Gö ls, H.; Schube , U. S. Unexpec ed
me al-media ed oxida ion o hyd oxyme hyl g oups o coo dina ed ca boxyla e g oups
by bis-cyclome ala ed i idium(III) cen e s. New J. Chem. 2010, 34, 2622-2633.
131
(23) Pomposo, J. A.; Colmene o, J.; Kohlb eche , J.; A be, A.; Sanchez-
Sanchez, A. E icien syn hesis o single-chain globules mimicking he mo phology
and polyme ase ac i i y o me alloenzymes. Mac omol. Rapid Commun. 2015, 36,
1592-1597.
(24) Robles-He nandez, B.; González, E.; Pomposo, J. A.; Colmene o, J.;
Aleg ía, A. Wa e dynamics and sel -assembly o single-chain nanopa icles in
concen a ed solu ions. So Ma e 2020, 16, 9738-9745.
(25) Chen, Z.; Jiang, J.; Zhao, Q.; An agg ega ion-induced phospho escen
emission-ac i e i idium(III) complex o luo ide anion imaging in li ing cells. J.
O ganome . Chem. 2021, 932, 1211644.
(26) Di, L.; Xing, Y.; Yang, Z.; Qiao, C.; Xia, Z. Sens. Pho os able
agg ega ion-induced emission o i idium(III) complex ealizing obus and high-
esolu ion imaging o la en inge p in s. Ac ua o s B Chem. 2023, 375, 132898.
(27) Twil on, J.; Le, C.; Zhang, P.; Shaw, M. H.; E ans, R. W; MacMillan, D.
W. C. The me ge o ansi ion me als and pho oca alysis. Na . Re . Chem. 2017, 1,
0052.
(28) S ie h-Kal ho , F.; James, J J.; Tede s, M.; Pi ze , L.; Glo ius, F, Ene gy
ans e ca alysis media ed by isible ligh : p inciples, applica ions, di ec ions. Chem.
Soc. Re . 2018, 47, 7190-7202.
(29) Shon, J.-H.; Kim, D.; Ra hnayake, M. D.; Si el, S.; Wea e , J.; Tee s, T.
S. Pho o edox ca alysis on unac i a ed subs a es wi h s ongly educing i idium
pho osensi ize s. Chem. Sci. 2021, 12, 4069-4078.
(30) a) Popla a, S.; T ös e , A.; Zou, Y.-Q.; Bach, T. Recen ad ances in he
syn hesis o cyclobu anes by ole in [2+2] pho ocycloaddi ion eac ions. Chem. Re .
2016, 116, 9748-9815. b) Du, J.; Yoon, T. P. C ossed in e molecula [2+]
cycloaddi ions o acyclic enones ia isible ligh pho oca alysis. Chem. Soc. 2009, 131,
14604-14605. c) Pagi e, S. K.; Hossain, A.; T aub, L.; Ke es, S.; Reise , O.
Pho osensi ized egioselec i e [2+2]-cycloaddi ion o cynnama es and ela ed alkenes.
Chem. Commun. 2017, 53, 12072-12075. d) Zhao, J.; B osme , J. L.; Tang, Q.; Yang,
Z.; Houk, K. N.; Diaconescu, P. L.; Kwon, O. In amolecula c ossed [2+2]
pho ocycloaddi ion ho ugh isible ligh -induced ene gy ans e . J. Am. Chem. Soc.
2017, 139, 9807-9810. e) Lu, Z.; Yoon, T. P. Visible ligh pho oca alysis o [2+2]
s y ene cycloaddi ions by ene gy ans e . Angew. Chem., In . Ed. 2012, 51, 10329-
10332. ) Tanaka, K.; Iwama, Y.; Kishimo o, M.; Oh suka, N.; Hoshino, Y.; Honda, K.
Redox po en ial con olled selec i e oxida ion o s y enes o egio- and s e eoselec i e
c ossed in e molecula [2+2] cycloaddi ion ia o ganopho o edox ca alysis. O g. Le .
2020, 22, 5207-5211.
132
(31) Ro h, H. G.; Rome o, N. A.; Nicewicz, D. A.; Expe imen al and calcula ed
elec ochemical po en ials o common o ganic molecules o applica ions o single-
elec on edox chemis y. Synle 2015, 27, 714-723.
(32) P ie , C. K.; Rankic, D. A.; McMillan, D. W. C. Visible ligh pho o edox
ca alysis wi h ansi ion me al complexes: applica ions in o ganic syn hesis. Chem.
Re . 2013, 113, 5322-5363.
(33) Eady, S. C.; Maclness, M. M.; Lehne , N. Immobilized cobal
bis(benzenedi hiola e) complexes: excep ionally ac i e he e ogeneous elec oca alys s
o dihyd ogen p oduc ion om mildly acidic aqueous solu ions. Ino g. Chem. 2017,
56, 11565-11576.
(34) Mon al i, M.; C edi, A.; P odi, L.; Gandol i, M. T.; Handbook o
pho ochemis y. Taylo & F ancis G oup, LCC 2006, 9780429115387.
(35) a) Kawaji i, T.; Ka o, M.; Naka a, H.; Go o, R.; Aiba a, S.; Oh a, R.;
Fujioka, H.; Sajiki, H.; Sawama, Y. A oma ic aldehydes and ace als ia py idinium sal
in e media es. J. O g. Chem. 2019, 84, 3853-3870. b) Ch is ensen, S. H.; Olsen, E. P.
K.; Rosenbaum, J.; Madsen, R. Hyd o o myla ion o ole ins and educ i e
ca bonyla ion o a yl halydes wi h syngas o med ex si u om dehyd ogena i e
deca bonyla ion o hexane-1,6-diol. O g. Biomol. Chem. 2015, 13, 938-945. c)
Desp as, G.; Zamalee a, A. I.; Da de e , L.; Tissey e, C.; Magalhaes, J. G.; Ga ne ,
C.; De Waa d M.; Amigo ena, S.; Fel z, A.; Malle , J.-M.; Collo , M. H-Rubies, a new
amily o ed emi ing luo escen pH senso s o li ing cells. Chem. Sci. 2015, 6, 5928-
5937. d) Shmid , B.; Eliza o , N.; Be ge , R.; Höl e , F. Scope and limi a ion o he
Heck-Ma suda-coupling o phenol diazonium sal s and s y enes: a p o ec ing-g oup
economic syn hesis o phenolic s ilbenes. O g. Biomol. Chem. 2013, 11, 3674-3691. e)
Hussain, M. I.; Feng, Y.; Hu, L.; Deng, Q.; Zhang, X.; Xiong, Y. Coppe -ca alyzed
oxida i e di unc ionaliza ion o e minal unac i a ed alkenes. J. O g. Chem. 2018, 83,
7852-7859. ) Mu phy-Benena o, K. E.; Oli ie , N.; Choy, A.; Ross, P. L.; Mille , M.
D.; Th eshe , J.; Gao, N.; Hale, M. R. Syn hesis, s uc u e, and SAR o
e ahyd opy an-based LpxC inhibi o s. ACS Med. Chem. Le ., 2014, 5, 1213-1218.
(36) a) Naik, A.; Meina, L.; Zabel, M.; Reise , O. E icien ae obic Wacke
oxida ion o s y enes using palladium bis(isoni ile) ca alys s. Chem. Eu . J. 2010, 16,
1624-1628. b) Daw, P.; Pe akamse y, R.; Sa bajna, A.; Laha, S.; Ramapanicke , R.;
Be a, J. K. A highly e icien ca alys o selec i e oxida i e scission o ole ins o
aldehydes: abno mal-NHC-Ru(II) complex in oxida ion chemis y. J. Am. Chem. Soc.
2014, 136, 40, 13987-13990. c) Xu, J.; Zhang, Y.; Yue, X.; Huo, J.; Xiong, D.; Zhang,
P. Selec i e oxida ion o alkenes o ca bonyls unde mild condi ions. G een Chem.
2021, 23, 5549-5555.
133
(37) a) Gao, R.; Ho, D. G.; He nandez, B.; Selke, M.; Mu phy, D.; Dju o ich,
P. I.; Thompson, M. E. Bis-cyclome ala ed I (III) complexes as e icien single oxygen
sensi ize s. J. Am. Chem. Soc. 2002, 124, 14828-14829. b) Xu, Y.; Wang, X.; Song,
K.; Du, J.; Liu, J.; Miao, Y.; Li, Y. BSA-encapsula ed cyclome ala ed i idium
complexes as nano-pho osensi ize s o pho odynamic he apy o umo cells. RSC Ad .
2021, 11, 15323-15331. c) Jiang, X.; Peng, J.; Wang, J.; Guo, X.; Zhao, D.; Ma, Y.
I idium-based high-sensi i i y oxygen senso s and pho osensi ize s wi h ul along
iple li e imes. ACS Appl. Ma e . In e aces 2016, 8, 3591–3600. d) Ashen-Ga y, D.;
Selke, M. Single oxygen gene a ion by cyclome ala ed complexes and applica ions.
Pho ochem Pho obiol. 2014, 90, 257–274. e) McKenzie, L. K.; Sazano ich, I. V.;
Baggaley, E.; Bonneau, M.; Gue chais, V.; Williams, J. A. G.; Weins ein, J. A.; B yan ,
H. E. Me al complexes o wo-pho on pho odynamic he apy: a cyclome ala ed
i idium complex induces wo-pho on pho osensi iza ion o cance cells unde nea -IR
ligh . Chem. Eu . J. 2017, 23, 234-238. ) Xu, Y.; Wang, X.; Song, K.; Du, J.; Liu, J.;
Miao, Y.; Li, Y. BSA-encapsula ed cyclome ala ed i idium complexes as nano-
pho osensi ize s o pho odynamic he apy o umo cells. RSC Ad . 2021, 11, 15323-
15331. g) Maggioni, D.; Galli, M.; D’Al onso, L.; In e so, D.; Dozzi, M. V.; Si oni,
L.; Iannacone, M.; Collini, M.; Fe u i, P.; Ranucci, E.; D’Al onso, G. A luminescen
poly(amidoamine)-i idium complex as new single -oxygen sensi ize o
pho odynamic he apy. Ino g. Chem. 2015, 54, 544-553. h) Qin, W.-W.; Pan, Z.-Y.;
Cai, D.-H.; Li, Y.; He, L. Cyclome ala ed i idium(III)-complexes o mi ochond ia-
a ge ed combined chemo-pho odynamic he apy. Dal on T ans. 2020, 49, 3562. i)
Zamo a, A.; Vigue as, G.; Rod íguez, V.; San ana, M. D.; Ruiz, J. Cyclome ala ed
i idium(III) luminescen complexes in he apy and pho o he apy. Coo d. Chem. Re .
2018, 360, 34-76.
(38) a) Zhao, J.-L.; Jiang, X.-K.; Wu, C.; Wang, C.-Z.; Zeng, X.; Redshaw, C.;
Yama o, T. An unp eceden ed pho ochemical eac ion o an h acene-con aining
de i a i es. ChemPhysChem 2016, 17, 3217-3222. b) Jacob, J. P.; Gopalak ishnan, R.;
Mallia, R. R.; Vadakkan, J. J.; Unnik ishan, P. A.; P a hapan, S. D ama ic sol en and
concen a ion dependence in he eac ion o (an h acen-9-yl)me hanamines wi h
sui able elec on-de icien ace ylenes. Phys. O g. Chem. 2014, 27, 884-891. c)Adam,
W.; P ein, M. Dias e eoselec i e [4+2] cycloaddi ion o syngle oxygen in he
pho ooxygena ion o chi al naph hyl alcohols: e idence o a hyd oxy g oup-di ec ing
e ec . J. Am. Chem. Soc. 1993, 115, 3766-3767. d) Mondal, S.; Mondal, S.; Midya, S.
P.; Das, S.; Mondal, S.; Ghosh, P. Me ging pho oca aly ic C-O c oss-coupling o α-
oxyca bonyl-β-ke ones: es e i ica ion o ca boxylic acids ia a deca boxyla i e
pa hway. O g. Le . 2023, 25, 184-189.
(39) a) Na a ajan, P.; Vagiche la, V. D.; Vijayan, M. T.; A mild oxida ion o
deac i a ed naph halenes and an h acenes o co esponding pa a-quinones by N-
b omosuccinimide. Te ahed on Le . 2014, 55, 3511-3515. b) Pankhu s , J. R.; Cu cio,
140
Figu e 4.1. Miles ones in he de elopmen o pho o he apy and pho odynamic
he apy o e he pas cen u y.32-35
The expe imen epo ed by Meye -Be z is wo h men ioning, being a mo e
han a scien i ic cu iosi y, he applied on his own skin 200 mg o hema opo phy in and
documen ed he ex ended in lamma ion esponse upon exposu e o sunligh (in Figu e
4.1, he pic u e o F ied ich Meye -Be z cap u ed a e ha ing a walk) is he i s
documen desc ibing he e ec s o a po phy in sys em in humans: po phy ins a e now
he mo e ex ensi ely s udied pho osensi ize s o PDT applica ions. These seminal and
undamen al obse a ions pa ed he way o wha is now a consolida ed ield o esea ch
and, in a non-i ele an numbe o cases, an app o ed medical ea men o a a ie y
o malignan solid umo s.
Pho o he apy is he use o ligh in he ea men o a disease, while
pho odynamic he apy (PDT), which can be conside ed as a speci ic
pho ochemo he apy, in ol es a combina ion o he adminis a ion o a pho osensi ize
and he use o ligh o ea a ce ain pa hological condi ion. Mo e speci ically, PDT
can be de ined as: a he apeu ic modali y, which in ol es wo indi idually non- oxic
componen (a pho osensi ize and ligh ) ha a e combined o induce cellula and issue
e ec in an oxygen-dependen manne .34 PDT in ol ed he adminis a ion, usually
ei he o al, o opic, o in a enous, o a pho osensi ize (PS) molecule, which is

141
biodis ibu ed acco ding o he biology o he o ganism unde s udy and can, o no ,
selec i ely accumula e in o gans o in e es o he ea men . A e he PS being
abso bed, he applica ion o ligh induces he p omo ion o PS g ound s a e o i s exci ed
s a es and, as o e e y pho oca aly ic p ocess, longe -li ing exci ed s a es (usually he
iple s) induces oxida i e s ess wi hin he cellule in which he PS ha e been
in e nalized. This la e can be he esul an o he di ec eac ion o PS exci ed s a es
wi h a subs a e impo an o cell su i al ( ype-I eac ions), gene a ing eac i e
adicals which can p oduce oxygena ed p oduc s upon eac ion wi h cellula oxygen.
Al e na i ely, PS iple s a e can di ec ly eac wi h cellula oxygen, o ming single
oxygen and eac i e oxygen species (ROS), which a e hen capable o induce high
oxida i e s ess in he cell, inally degene a ing in o cell apop osis ( ype-II eac ions).
In Figu e 4.2, he subs an ial di e en mechanis ic pa hways wi h which PDT can
ac ua e in he cellula medium a e illus a ed. The p opo ion o hese wo ypes o
eac ion aking place elies on he p ope ies o he speci ic PS used, in all cases, PDT
is an oxygen-dependen p ocess whose undamen al p inciples ely on pho oca alysis
in he biological, s ill aqueous, en i onmen .
Figu e 4.2. Pic o ial ep esen a ion o he basic p inciples o PDT.34, 36, 37
4.1.2. T adi ional Pho osensi ize s o PDT
Ideal pho osensi ize s o PDT should possess a good lipophilic balance, o
simul aneously ensu e e icien biodis ibu ion and accumula ion in he umo al issues,
142
as well as high ligh abso p ion coe icien s, p e e ably a as-long-as-possible
wa eleng hs, high ISC e iciencies, negligible da k cy o oxici y and low
pho obleaching. A he da e, di e en PS ha e been deeply in es iga ed and widely
used o he apeu ic applica ions, and can be classi ied in h ee subg oups, o
gene a ions. Po phy ins like hema opo phy in, whose chemical s uc u e is shown in
Figu e 4.3, a e i s -gene a ion pho osensi ize s and a e adi ionally na u al occu ing
subs ance wi h high quan um yields o p oduce ROS.38
Figu e 4.3. S uc u es o commonly employed molecula a chi ec u es o i s and
second-gene a ion pho osensi ize o PDT wi h hei espec i e wa eleng h o ac i a ion. a)
hema opo phy in, b) p o opo phy in IX, c) chlo in e6, d) iodine-subs i u ed BODIPY, e) Lu-
exaphy in,39 ) modi ied aza-BODIPY, g) Ru[(dmb)2(IP-3T)]Cl2 (TLD1433).40
Hema opo phy ins and hei analogues ha e been ex ensi ely epo ed as
e ec i e PDT agen s since he e y i s epo s on he ield, howe e , wi h hei
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ex ended polycyclic a oma ic s uc u e low solubili y in wa e usually is o cen al
conce n.40 Second-gene a ion PS a e commonly subs ances designed o o e come hese
majo issues, as o chlo in e6 and p o opo phy in IX and Lu- exaphy in (shown in
Figu e 4.3), he basic s uc u e o i s -gene a ion pho osensi ize s we e modi ied o
imp o e c i ical ea u es like solubili y in wa e , wa eleng h o abso bance and
enhance ROS gene a ion quan um e iciency. Comple ely no el molecula s uc u es
as a iously unc ionalized 4,4-di luo o-4-bo a-3a,4a-diaza-s-indacenes (BODIPYs) as
well as wa e -soluble cyclome ala ed ansi ion-me al complexes (Figu e 4.3) can be
classi ied as second-gene a ion ca alys . In pa icula , BODIPY appea ed o be an
in e es ing molecula s uc u e o hei low pho obleaching deg ees, highe s abili y in
biological en i onmen , low da k- oxici y and high ligh abso p ion coe icien s,
howe e , due o he high luminescence hey usually p esen low single oxygen
gene a ion quan um yields. Rega ding cyclome ala ed ansi ion-me al complexes, hey
commonly p esen ROS e iciency o compa able en i y han ha o ph halocyanine
and po phy i ic sys ems and, helped by he ich chemis y o hei elec onic exci ed
s a es, usually mani es high pho o oxci i ies.40,41 Fo hese easons, hey ha e been o
pa icula in e es as sys ems ha can enable PDT in hypoxic condi ion and a low
oxygen ension issues, as in he case o TLD1433, he u henium(II) polypy idyl
complex Ru[(dmb)2(IP-3T)]Cl2 (Figu e 4.3) being he i s o ganome allic o each
human clinical ials, which exploi s long-li ing iple s a es o ei he single -oxygen
sensi iza ion and o ini ia e in acellula cy o oxic adical pa hways.40
4.1.3. Nanos uc u es in PDT
As a gene al aspec , he induc ion o e ec i e and selec i e des uc ion o
damaged and cance ous cells while o bea ing he su ounding heal hy issues is o
pi o al impo ance o PDT. Despi e he p omising esul s and p og ess egis e ed o e
he las cen u y, PDT is cu en ly an app o ed clinical solu ion only o e y speci ic
cases o umo s, which gene ally mus be supe icial and la , o accessible wi h
endoscopes o ensu e good illumina ion o he a ec ed egion. PDT is no applicable
o highly di used, me as a ic umo s, as is no possible o illumina e he en i e body o
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a pa ien wi h he echnology we cu en ly dispose o . The same can be said o solid,
deep-sea ed umo s, in which ligh pene a ion, and so PSs ac i a ion, is o majo
cons ain .42 Selec i i y o cance issue is usually achie ed by aking ad an age o he
abno mal physiology o his la e , which commonly p esen ei he highe
ascula iza ion, poo lympha ic d ainage o dec eased pH, all ea u es which can be
exploi ed o designing sma e PSs capable o selec i e accumula ion wi hin he
umo . In his sense, he use o nanopa icles o PDT e iciency imp o emen is a
pa icula ly p omising app oach o se e al easons. An impo an bene i a ising om
he use o a nanos uc u e o PSs deli e y o PDT is gi en by he possibili y o
inducing enhanced pe meabili y and e en ion (EPR) e ec . Al e ed ascula iza ion o
umo al issue no mally esul s in abno mal in e naliza ion o ci cula ing species in he
blood sys ems o he a ec ed o ganisms and, when he size o he nanos uc u es
employed o he d ug deli e y is app op ia ely selec ed, his abno mal physiology can
lead o selec i e accumula ion o he bigge objec s wi hin he umo ( e en ion).43 This
way, he use o nano-sized objec s has been exploi ed o mo e a s ep o wa d a mo e
selec i e PDT, pe mi ing a mo e si e-speci ic accumula ion o he pho oac i e species,
u he ly imp o ing he cellula up ake, biodis ibu ion, pha macokine ics and
minimizing he unwan ed side e ec s a ising om sca cely selec i e accumula ion.44
O he bene i s o using nanopa icles in PDT include, la ge su ace- o- olume a ios,
which always co esponds o enhanced e icacy o he deli e ed species in he a ge
cells, he p ema u e deli e y o he PSs is also p e en ed, wi h his being especially
ue when a non-biodeg adable, ine nanopa icle is used as a passi e ca ie and, no
less impo an ly, nanopa icles can be p epa ed in a a ie y o ashions, opologies,
dimensions and wi h he possibili y o almos inexhaus ible ways o chemical
unc ionaliza ion o imp o ing biodis ibu ion and PSs pe o mance.42-45 Conce ning
ino ganic-based sys ems, gold nanopa icles (AuNPs) ha e demons a ed o be
employable as e ec i e passi e PSs nanoca ie s o PDT-applica ions. Apa om
being usually s able, ha d sys ems which p esen op imal biocompa ibili y, wi h his
explaining i s wide use in nanomedicine mo e in gene al, AuNPs can be easily
unc ionalized ei he co alen ly o non-co alen ly ia he easily accessible me allic
gold- hiola e chemis y. In Figu e 4.4 a e shown wo examples o he di e en
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app oaches which can be ollowed o inco po a ing a PS in o AuNPs, which can be
ei he co alen , as epo ed by Russel e al. in he unc ionaliza ion o PEGyla ed
AuNPs wi h zinc(II)-ph halocyanine con aining hiola e moie ies,24 o non-co alen ly,
exploi ing s ong ionic in e ac ions be ween opposi e cha ged NPs and PS, as epo ed
by Pé ez-Ga cía e al. in he p epa a ion o posi i ely cha ged, wa e -soluble AuNPs
loaded wi h anionic po phy ins.46
Figu e 4.4. Illus a ion o wo exempli ica i e s a egies owa ds PDT-ac i e
AuNPs. On op, co alen unc ionaliza ion o he su ace o me allic gold nanopa icles wi h
hyd ophilic PEG-based pendan s and hiola e-bea ing zinc(III)ph halocyanine.47 On he
bo om, he wa e -soluble pho oac i e AuNPs sys em assembled ia coulombic o ces
be ween py idinium species and anionic po phy ins.46
Analogously, nanos uc u ed silica also o e s a ple ho a o possibili ies in
e ms o i s applica ion in nanomedicine. I s de i a i e nanos uc u es a e well-known
o hei biocompa ibili y and he possibili y o subsequen unc ionaliza ion, as well
o he usually high colloidal s abili y o he inal p oduc . Silica-based nanopa icles

146
ha e al eady been applied o he ab ica ion o PDT-ac i e ma e ials, as PSs can be
inco po a ed in o mesopo ous silica nanopa icles, o used o chemical modi ica ion
o silica nano-objec s and, no ably, silica can be used o coa p e-exis ing nanopa icles,
hanks o he possibili y o laye -by-laye deposi ion.48,49 Amongs he di e en
oppo uni ies p o ided by ino ganic-based ma e ials, i on oxide nanopa icles (IONPs)
a e wo h men ioning, due o he possibili y o e ed by he applica ion o local magne ic
ields, ei he s a ic o di ec ing o he desi ed si e o ac ion o al e na ing o inducing
magne ic hype he mia, IONPs gained special a en ion in nanomedicine in ecen
imes50,51 and, o wha conce ns PDT, hey ha e al eady been demons a ed o expand
he possibili ies o mul i-di ec ed app oaches (coupling PDT wi h magne ic induced
hype he mia) when included wi h PSs in liposomes and cell-mimicking s uc u es.52
Nanopa icles can o en gua an ee augmen ed PSs solubiliza ion in he
complex, aqueous medium o he biological sys ems. E.g., he use o polyme ic
micelles o ca y s ongly hyd ophobic PSs by means o hei non-co alen
encapsula ion wi hin he amphiphilic nanopa icles and / o co alen ancho ing in o he
hyd ophobic co e ha e been demons a ed in se e al applica ions, including Plu onic,
PEG-based lipids, pH- esponsi e Poly(N-isop opyl ac ylamide) (PNIPAM)-based
micelles and poly-ion complex micelles, o ci e a ew.53 As a gene al beha io , he
micella co e is esponsible o he d ug-ca ying capabili y o hese polyme ic nano-
ec o s, since a a ie y o hyd ophobic d ugs can be inco po a ed in o he co e by non-
co alen in e ac ions. Fo his eason, PS a e usually inco po a ed in o polyme ic
micelles h ough hyd ophobic in e ac ions wi h he segmen ha o ms he micella
co e, by means o physical en apmen . PS can also be ca ied h ough co alen bonds
be ween unc ional g oups o he PS and penden g oup o hyd ophobic segmen s o
he polyme ic sca old used o he sup amolecula assembly o ma ion. I is wo h
men ioning ha he s abili y o he physical apping o he PS depends on he
magni ude o he hyd ophobic in e ac ions be ween he PS and he hyd ophobic pa o
he copolyme .54
Amongs all he possibili ies o e ed by he cons uc ion o nano-objec s o
con olled s uc u e and shape, single-chain polyme ic nanopa icles (polyme ic
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SCNPs) p o ide an in e es ing sca old o medicinal chemis y. Thanks o hei
de ined shape, unable mo phology esembling he one o enzymes and na u ally
occu ing p o eins, as well as he ease o unc ionaliza ion, good wa e -solubili y, and
ul a-small dimensions (2-20 nm), SCNPs al eady ha e been p o ed o be e icien
nanoca ie s o d ug-deli e y applica ions and imaging. Due o he by-de ini ion
unimolecula cha ac e , SCNPs a e usually mo e dilu ion- esis an han micelles and
liposomes, whose majo cons ain is indeed coalescence in he bloods eam o en
leading o PS agg ega ion, which is usually de imen al o he pha macokine ics and
PSs e icacy o e ime. Also due o hei ul a- educed dimensions, SCNPs a e usually
highly compa ible wi h he in acellula space.55,56 In addi ion, he collapse/ olding o
indi idual polyme ic chains allows he o ma ion o indi idual pocke s in which he
pho oac i e uni s can be isola ed om hemsel es, which is gene ally associa ed wi h
he enhancemen o hei op ical p ope ies, hence esul ing in augmen a ion o ROS
gene a ion in he biological media. In a seminal, ecen wo k epo ed by Meije e al.
he use o a no el class o po phy in-con aining, wa e -soluble SCNPs is p oposed as
inno a i e sca old o aqueous pho osensi iza ions.57 The au ho s de eloped he
pho oac i e single-chain nanoca ie by means o a pos -polyme iza ion
unc ionaliza ion o an ul a-high molecula weigh polyme ic p ecu so
Poly(pen a luo o s y ene), while le eac ing speci ic amoun o he po phy in-based
pho osensi ize and Je amine, con e ing wa e -solubili y o he inal p oduc .
The eadily p epa ed pho oac i e polyme esul ed o sel -assemble in aqueous
en i onmen in o he collapsed/ olded unimolecula s uc u e we e hen e alua ed o
hei easibili y in aqueous pho osensi iza ion by means o a spec opho ome ic
in es iga ion, bo h in he UV-Vis ange o he de e mina ion po phy in agg ega ion-
ex en and in a- ed (IR) emission spec oscopy o single -oxygen gene a ion abili y
es ima ion.57 In a la e wo k, Liu e al. de eloped analogous SCPNs-based sys ems
bea ing pho oca aly ic po phy ins, which we e success ully in oduced in o he
lysosomal egion o HeLa cells ia endocy osis.58 In e es ingly, he au ho s epo ed no
signi ican changes in he emission spec a o he po phy ins upon cell in e naliza ion,
indica ing minimal agg ega ion o modi ica ion o he unc ional uni . The po phy in
based SCPN induced signi ican cell dea h a e i adia ing he cells wi h λ exc max = 403
148
nm ligh , which ag ees wi h hei abili y o gene a e cy o oxic 1O2 single oxygen. The
ema kable single -oxygen capabili y o he PS-loaded SCNP, in compa ison o he
ee-po phy in sys em, could no only be due o an augmen ed s abiliza ion in wa e o
he PS, bu also o he ac ha SCNPs possess a compac bu a he open and lexible
s uc u e, allowing o he eac an s ( iple molecula oxygen) and he p oduc s (ROS)
o di use easily in and ou he nanopa icles. To summa ize, wi h a pho oac i e,
po phy in-con aining SCNPs being e alua ed o PDT-e ec in a p ope biological
sys em, al hough hese pionee ing wo ks demons a e ha he inclusion o a
hyd ophobic, highly agg ega ing PDT-sui able pho osensi ize wi hin he a chi ec u e
o a wa e -soluble SCNP cons i u e a alid s a egy o ROS-gene a ing dyes o
he apeu ic applica ions, s ill a lo o e o ha e o be pu o e ealing he hidden
po en ial o pho oca aly ic SCNPs o be exploi ed as e icien , las gene a ion PDT-
nanod ugs.
4.2. Objec i es
When planning an encapsula ion s a egy, sup amolecula app oaches
cons i u e a alid op ion owa ds he o mula ion o mo e pe o ma i e PSs. The ypes
o agg ega es a ising om di e en nonco alen in e ac ions can con e o ma e ials
in iguing pho ochemical p ope ies, e y di e en om he single cons i uen s aken
alone.59 In Na u e, pho oca aly ic p ocesses a e enabled in wa e by aminoacidic single-
chain nanopa icles, which we call p o eins, whose p ecise olding allows he
s abiliza ion and e icien unc ioning o pho osensi izing molecules in he chemically
complex, aqueous, biological media. As i happens o he mos abundan
pho oca alyzed eac ion on Ea h, he chlo ophyll-based pho osyn hesis, he ca e ul
assemble o p o eins and pigmen s (o en hyd ophobic molecules) is equi ed o he
ca aly ic p ocess o ake place. E.g., highe plan s pho osys em I (PSI) is composed,
amongs o he s, by he unique assembly o ou di e en ligh -ha es ing p o eins
(LHCI), which welcome and enclose in hei olded s uc u e a o al o 165 chlo ophylls
and 5 addi ional elec on-ca ying co ac o s as phylloquinones and Fe-S clus e s, as
epo ed in he s uc u e shown in Figu e 4.5.60,61
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Figu e 4.5. C ys al s uc u es o pa o he cons i u i e uni s o he PSI om Pisum
Sa i um. Speci ically, he con ac egion o Lhca1 (g een) and Lhca4 (g ey) is shown. The
magen a solid line ollows schema ically he backbone o he p o einic single-chain uni
Lhca1. Enclosed chlo ophylls a e depic ed in blue i bound o LHCA monome s, in ed when
p esen a linke unc ion.60
Due o he ex ended elec on delocaliza ion in he e apy olic s uc u e, i is
wo h no ing ha ph halocyanines a e capable o speci ically in e ac h ough π-π
in e ac ions wi h he a oma ic ing sys ems o p o eins, o he poin ha ha e been
epo ed o be able o modula ion o unwan ed amyloid agg ega ion o p o eins.62
O ganic ch omopho es like an h acene, pe ylene e c. a e, in ac , knowingly p one o
agg ega ion and, al hough hei p ope ies in he condensed s a e a e di icul o p edic
and con ol, se e al s a egies o une hei assembly a e o en desi able when hinking
on ad anced applica ion o hei op oelec onic unc ionali ies. The use o me al
o ganic amewo ks,63,64 co-assembly wi h biomac omolecules,65 and hos -gues
encapsula ion,66 a e some o he possible app oaches owa d he design o unable-
agg ega ion o ph halocyanine PSs-based ma e ials.
The p esen wo k aimed a he a ional design and p epa a ion o imp o ed
amphiphilic single-chain polyme nanopa icles (SCNP) o imaging and
pho odynamic he apy (PDT) in zeb a ish emb yo xenog a s. As desc ibed in he
p e ious chap e s, SCNPs a e ul a-small polyme ic nanopa icles wi h sizes simila o