Review Article The Chemistry of Bioconjugation in...

Review ArticleThe Chemistry of Bioconjugation in Nanoparticles-BasedDrug Delivery System

Karolina Werengowska-CieTwierz1 Marek WiVniewski12

Artur P Terzyk1 and Sylwester Furmaniak1

1Physicochemistry of Carbon Materials Research Group Faculty of Chemistry Nicolaus Copernicus University in TorunGagarin Street 7 87-100 Torun Poland2INVEST-TECH RampD Center 32-34 Plaska Street 87-100 Torun Poland

Correspondence should be addressed to Sylwester Furmaniak sfchemumkpl

Received 10 November 2014 Accepted 1 February 2015

Academic Editor Mohindar S Seehra

Copyright copy 2015 Karolina Werengowska-Ciecwierz et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Nanomedicine is generally the application of nanotechnology to medicine The term nanomedicine includes monitoringconstruction of novel drug delivery systems and any possible future applications of nanotechnology and nanovaccinology In thisreview themost important ligand-nanocarrier and drug-nanocarrier bioconjugations are describedThe detailed characterizationsof covalently formed bonds between targeted ligand and nanocarrier including amide thioether disulfide acetyl-hydrazone andpolycyclic groups are described Also the coupling of small elements and heteroatoms in the form of R-X-R the ldquoclick chemistryrdquogroups is shown Physical adsorption and chemical bonding of drug to nanocarrier surface involving drug on the internal or externalsurfaces of nanocarriers are described throughout possibility of the formation of the above-mentioned functionalities Moreoverthe most popular nanostructures (liposomes micelles polymeric nanoparticles dendrimers carbon nanotubes and nanohorns)are characterized as nanocarriers Building of modern drug carrier is a new method which could be effectively applied in targetedanticancer therapy

1 Introduction Nanotechnology andNanomedicine

Nanotechnology has led to a junction of different fieldsincluding chemistry biology applied physics optics compu-tational analysis andmodeling andmaterials science Recentadvances in the physical sciences have provided the abilityto analyze and manipulate structures at nanometer scalesand this has been accompanied by advances in molecularmodeling and computational science Using them one canpredict bymodeling and simulation the behavior of biologicalstructures in (the most important for human) aqueoussolution [1] The purpose of nanomedicine is the same as thatof medicine to diagnose as accurately and early as possibleand to treat as effectively as possible It means that side effectsshould be avoided [2] Human health-care nanotechnologyresearch can definitely result in immense health benefits [3]

Therefore there is increasing funding for nanotechnology allover the world [4] Estimates of the impact from advancesemerging from nanotechnology developments over the next15 to 20 years have been estimated to be approximately$1 trillion by studies conducted at the National ScienceFoundation [3]

Generally nanomedicine may be defined as the monitor-ing reparation construction and control of human biologi-cal systems at themolecular level To do this nanodevices andnanostructures are applied [3] The prefix ldquonano-rdquo means theuse of materials of which at least one of their dimensions is inthe scale range 1ndash100 nm [5]

There are numerous methods for drugs delivering intoorganisms They include oral transdermal transepithelialand intravenous delivery All of these methods could befulfilled using nanostructures as drug containers Differentforms of drug nanocontainers as polymeric nanoparticles

Hindawi Publishing CorporationAdvances in Condensed Matter PhysicsVolume 2015 Article ID 198175 27 pageshttpdxdoiorg1011552015198175

2 Advances in Condensed Matter Physics

Classical therapy Targeted therapy

Cancer cell Endothelial cell

Nanocarrier Antibody nanocarrier

Highconcentration

Lowconcentration

ligand to receptorHigh affinity ofEPR effect

Figure 1 The mechanism of drug delivery in classical and targeted therapy (the figure is based on [12 199])

liposomes magnetic nanoparticles and carbon nanostruc-tures are extensively studied [4ndash10] All theseDDSs have beenreported to have huge biomedical potential

Nanomedicine is a rapidly developing research areain biological and medical science and the development ofnanomedicine research requires collaboration among dif-ferent scientists physicians engineers molecular biologistsmaterial scientists chemists and so forth [8] Applicationsof nanotechnology in medicine are especially promisingand areas such as disease diagnosis drug delivery targetedat specific sites in the body (see the next paragraph) andmolecular imaging are intensively investigated and someproducts are undergoing clinical trials Nanotechnology playsan important role in therapies of the future as ldquonanomedicinerdquoby lowering doses required for efficacy as well as increasingthe therapeutic indices and safety profiles of new therapeutics[9] The therapeutics are delivery systems in the nanometersize range containing encapsulated dispersed adsorbed orconjugated drugs and imaging agents Selected nanoscalesystems including liposomes micelles nanoemulsionsnanoparticulate systems (drug nanoparticles polymer-lipid- carbon- and ceramic-based albumin and nanogels)and dendrimers are used for drug delivery and imaging [9]

Within the next 5 to 10 years nanomedicine will addressmany important medical problems by using nanoscale-structured materials and simple nanodevices that can bemanufactured today Many approaches to nanomedicinebeing carried out today are already close enough to fruitionthat it is fair to say that their successful development isalmost inevitable and their subsequent incorporation intovaluable medical diagnostics or clinical therapeutics ishighly likely and may occur very soon Among them Fre-itas Jr [4] mentioned immunoisolation gated nanosievesultrafast DNA sequencing fullerene-based pharmaceuticalsnanoshells single-virus detectors tectodendrimers radio-controlled biomolecules and biologic robots [4] Flynn andWei [10] mentioned also the importance of commercializa-tion of nanomedicine

In the next chapter we present short review on one ofthe practical applications of nanotechnology inmedicineWefocus on the application of nanoparticles as DDS

2 Targeted Anticancer Therapy

A deeper understanding of the molecular events leading totumor formation invasion angiogenesis and metastasis hasprovided a newmechanistic basis for drug discovery targetedanticancer therapy By specifically blocking the molecularpathways implicated in the pathogenesis of cancer targetedanticancer agents are expected to alter the natural courseof the disease and at the same time to offer an enhancedtherapeutic index over traditional cytotoxic agents

Targeted anticancer therapy must fulfill few requestsAnticancer drugs should be delivered to the cancer cellswith minimal activity loss The drug should kill selectivelythe cancer cells The release of drug active form must beof course controlled [11] Simultaneously only the minimaldoses of chemotherapeutic agent should be administeredduring the targeted therapy (doses should be smaller thanthose in the traditional chemotherapy) The therapy shouldalso minimize different possible side effects [11 12]

Currently drugs are usually delivered inside or outsideof nanocarriers via the passive or selective mechanism inconventional or targeted therapy respectively [12] (Figure 1)The traditional mechanism is connected with a drug aggre-gation process (in form of carrier drug) inside cancer tissuesvia enhanced permeability and retention effect (EPR) [13ndash15] This method is based on the abnormal structure ofblood vessels near tumor Thus the drug reaches easily thetissues near cancer cells [9 14] The most popular drugsused during conventional treatment are doxorubicin [16]paclitaxel [17] methotrexate [18] hexamethylmelamine [19]and gemcitabine [20] and drugs based on platinum [21] likecisplatin (DDP) or carboplatin (Figure 2) The linking ofdrugs to nanocarriers is described in this paper together withprogressive activity after linking with antibody

Advances in Condensed Matter Physics 3

OHOH

O

ONH

O

OO

AcO O OH

OHAcO

(a)

OO

O OH O

O

OH

OH

OH

OCH3

NH2

CH2OH

(b)

N N

NN N

H2N

NH2NH

OH

OH

O

O

O

(c)

N

N

O O

F

FOH

HO

NH2

(d)

PtOCO

OCO

H3N

H3N

(e)

PtCl

ClH3N

H3N

(f)

N

N

N

N

N

N

(g)

Figure 2 Chemical structures of drugs (a) paclitaxel (b) doxorubicin (c) methotrexate (d) gemcitabine (e) carboplatin (f) cisplatin and(g) hexamethylmelamine

The targeted anticancer therapy uses systems composedof the ligands connected to the carriers of chemotherapeuticagent Thus constructed structure facilitates the bioconjuga-tion with appropriate receptors of cancer cells The targetedanticancer therapy in contrast to the conventional treatmentsupports the overexpression of cancer cell receptors and theaffinity of ligand to receptor [12 14 15]Themain advantage oftargeted treatment is the delivery of chemotherapeutic agentsto the most resistant cancer cells and longtime circulationinside them Application of this method guarantees highconcentration of drug inside the tumor Moreover drugcannot be released back to the blood The main factordetermining the type of used ligand is immunogenicity [13]

3 Nanoparticles as Drug Delivery System

31 The Importance of Proper Drug Delivery The Role ofNanoparticles The major practical purpose of nanotechnol-ogy in medicine is the application of nanoparticles in DDS[22] The reason of this is that during the past two decades

researchers involved in the development of pharmaceuticalshave understood that drug delivery is a fundamental part ofdrug development and a wide range of DDS has thus beendesigned [3] It is also very important that at present 95of all new potential therapeutics have poor pharmacokineticsand biopharmaceutical properties [9] The therapeutic indexof nearly all drugs currently being used would be improved ifthey were more efficiently delivered to their biological targetsthrough appropriate application of nanotechnologies Somedrugs that have previously failed clinical trials might also bereexamined using nanotechnological approaches [22]

What are the requirements for an effective and safe drugFor example in anticancer therapy theremust be an adequatedrug concentration in the body to allow for an effectivedose at the tumor site The target must be strongly inhibitedwith the function essential for tumor cell viability The drugmust have a high toxicity toward the tumor or a favorabletherapeutic windowMany authors have delivered a variety ofdrugs such as hydrophilic and hydrophobic drugs proteinsvaccines and biological macromolecules using nanoparticles


as carriers For the delivery of antigens for vaccination[3] three main types of gene delivery systems have beendescribed viral vectors nonviral vectors (in the form ofparticles such as nanoparticles liposomes or dendrimers)and the direct injection of genetic materials into tissues usingso-called gene guns [3] Nanostructured architectures arepromising candidates that will enable targeted delivery ofnovel drug compounds Nanoscale drug delivery mechanismhas effects on continuous drug release and intracellular entrycapability Moreover it minimizes side effects and allows forthe direct treatment of the cause of the disease rather than thesymptoms of the illness Generally nanoparticles [14 23 24]

(i) have advantage over larger microparticles becausethey are better suited for intravenous delivery

(ii) have been highly exploited for controlled drug releaseand site-specific drug targeting

(iii) have shown promising results in the case of site-specific drug targeting for treating various diseasesincluding cancer human immunodeficiency virusinfection and central nervous system disorders

(iv) have a higher surface to volume ratio as comparedwith bulk material and therefore the dose and fre-quency of administration would be reduced henceincreasing patient compliance recently it was shownthat solid lipid doxorubicin loaded nanoparticles havepotential to acne

(v) have the additional advantage of prolonged circula-tion in the blood which would facilitate extravasa-tion and passive targeting (nanoparticles made withhydrophilic polymers)

(vi) avoid opsonization with particle size less than 100 nm(hydrophilic nanoparticles) These systems prolongthe duration of action as well as increasing thetargeting of the drug to specific sites [25]

Multifunctional nanodelivery systems could combinetargeting diagnostic and therapeutic actions There arealready an astonishing number of emerging applicationsThese purposes either take advantage of the unique propertiesof nanoparticles as drugs or components of drugs per se orare designed for new approaches to controlled release drugtargeting and salvage of drugs with low bioavailability [3]

As mentioned above the success of a therapy dependson the drug delivery method Its importance is exemplifiedby the presence of more than 300 companies based on theUnited States involved with developing drug delivery plat-forms In addition to the commonly used oral and injectionroutes drugs can also be administered through other meansincluding transdermal transmucosal ocular pulmonary andimplantation delivery The mechanisms used to achievealternative drug delivery typically incorporate one or moreof the following materials biologics polymers silicon-basedmaterials carbon-based materials or metals [3]

In this paragraph we will focus mainly on carbonnanomaterials in drug delivery reporting the results ofnew findings The applications of biologic structures poly-mers dendrimers silicon-based structures and some carbon

materials in drug delivery were reviewed in [24] In [26]the review on nanoshells carbon nanotubes dendrimerssuperparamagnetic nanoparticles and liposomes applied incancer therapeutics is presented All of these nanotechnologyplatforms can be multifunctional and so they are frequentlynamed ldquosmartrdquo or ldquointelligentrdquoThe authors raise awareness ofthe physiological challenges for the application of these ther-apeutic nanotechnologies in light of some recent advances inour understanding of tumor biology [26]

When drugs and imaging agents are associated withnanoscale carriers their volumes of distribution are reducedNanoscale DDS also has the ability to improve the pharma-cokinetics and increase biodistribution of therapeutic agentsto target organs which will result in improved efficiencyDrug toxicity is reduced as a consequence of preferentialaccumulation at target sites and lower concentration inhealthy tissues Nanocarriers have been designed to targettumors and inflammation sites that have permeable vascu-lature Targeting and reduced clearance increase therapeuticindex and lower the dose required for efficacy Deliverysystems have been shown to increase the stability of awide variety of therapeutic agents such as small hydropho-bic molecules peptides and oligonucleotides Nanocarrierscomposed of biocompatible materials are investigated as safealternatives to existing vehicles that may cause hypersensi-tivity reactions and peripheral neuropathy [9] A numberof additional obstacles can be overcome with various novelapplications of nanodrug delivery Many drugs are not verysoluble making it difficult to administer therapeutic dosesThese compounds can be ldquosolubilizedrdquo by formulating theminto crystalline nanosuspensions that are stabilized by surfac-tants or by combining them with organic or lipid nanopar-ticles that keep them in circulation for longer periods If anefficacious compound has a short half-life in the circulationits stability can be increased tremendously by encasing itwithin for example nanosized liposome as a drug carrier Inthe case of cancers for example of central nervous systemmany drugs have difficulty in crossing the blood-brain barrierto attack the tumor Drug-loaded nanoparticles are ableto penetrate this barrier and have been shown to greatlyincrease therapeutic concentrations of anticancer drugs inbrain tumors The best way to increase the efficiency andto reduce the toxicity of a drug is to direct it into its targetand maintain its concentration at the site for a sufficienttime for therapeutic action to take effect [22] The majorityof solid tumors exhibit a vascular pore cutoff size between380 and 780 nm [9] although tumor vasculature organizationmay differ depending on the tumor type its growth rateand microenvironment Therefore particles need to be of asize much smaller than the cutoff pore diameter to reachto the target tumor sites By contrast normal vasculatureis impermeable to drug associated carriers larger than 2 to4 nm compared to free unassociated drug molecules Thisnanosized window offers the opportunity to increase drugaccumulation and local concentration in target sites such astumor or inflamed sites by extravasations and significantly toreduce drug distribution and toxicity to normal tissues [9]

Ideal DDS can be achieved by creation of materialsundergoing no chemical changes and satisfying the demands


of biodegradability and biocompatibility of the nanoparticlescarrier the rate of biodegradation of the carrier and therelease dynamics of the drug [15 27] For example for so-called passive targeting to be successful the nanocarriersneed to circulate in the blood for extended times so thatthere will be multiple possibilities for the nanocarriers topass by the target site Nanoparticulates usually have shortcirculation half-lives due to natural defense mechanismsof the body to eliminate them after opsonization by themononuclear phagocytic system (also known as reticuloen-dothelial system) Therefore the particle surfaces need tobe modified to be ldquoinvisiblerdquo to opsonization A hydrophilicpolymer such as polyethylene glycol (PEG) is commonlyused for this purpose because it has desirable attributessuch as low degree of immunogenicity and antigenicitychemical inertness of the polymer backbone and availabilityof the terminal primary hydroxyl groups for derivatiza-tion PEG-grafted liposomes in the size range of 70 to200 nm containing 3 to 7mol methoxy-PEG-2000 graftedto distearoyl phosphatidylethanolamine (DSPE) or dipalmi-toyl phosphatidylethanolamine showed extended circulationhalf-lives of 15 to 24 hours in rodents and up to 45 hours inhumans whereas non-PEGylated liposomes had half-lives of2 hours [15 27]

Nanocarriers typically consist of macromolecular mate-rials with the active principle either dissolved within apolymeric matrix entrapped inside lipid encapsulated oradsorbed onto surfaces of particles Accordingly they canbe classified into mainly two types nanocapsules andnanospheres The former are vesicular systems in whichdrug molecules are surrounded by a membrane whereas thelatter are matrix systems with the drug molecules dispersingthroughout [2] Though the technology is still young morethan 1000 nanopharmaceutical patents have been issued bythe US Patent and Trademark Office (US PTO) duringthe period 1999ndash2008 [2] Nanotechnology-based methodsof synthesis are most commonly developed on the basis oftwo rational designs either top-down or bottom-up engi-neering of individual components The top-down processinvolves starting with a larger object and breaking it up intonanostructures through etching grinding or ball millingThe process can be accelerated by addition of chemicalsor using laser Microscale or macroscale manufacturinglike silicon microfabrication and photolithography is oftenaccomplished as top-down process However the methodis time-consuming and frequently generates considerablybroader particle size distribution The bottom-up techniquerefers to synthesis based on atom-by-atom or molecule-by-molecule arrangement in a controlled manner The processtakes place through controlled chemical reactions in eithergas or liquid phase resulting in nucleation and growth ofnanoparticles Bottom-up techniques (like supercritical fluidantisolvent techniques precipitation methods etc) createheavily clustered masses of particles that do not breakup on reconstitution [2] All preparation methods high-pressure homogenization complex coacervation coprecipi-tation salting-out nanoprecipitation solvent emulsification-diffusion supercritical fluid rapid expansion of supercrit-ical solutions supercritical antisolvent precipitation and

self-assembly methods were in detail described in [2]Among nanoparticles they describe polymeric nanoparticlessolid lipid nanoparticles magnetic nanoparticles metal andinorganic nanoparticles quantum dots polymeric micelles(PMs) phospholipid micelles and colloidal nanoliposomes

32 Drug Delivery Systems The examples of recent appli-cation of nanoscale systems for drug delivery are shown inFigure 3 We focus on solid nanomaterials most often usedas nanocarriers These carriers after joining some ligandsandor drugs can be used in designing of the systems fortargeted therapies as described in the following sections

Liposomes and lipids have been used as DDS since 1960Liposomes are defined as vesicles in which an aqueousvolume is entirely surrounded by a phospholipid membraneLiposome size can vary from30 nmup to severalmicrometersand can be uni- or multilamellar [9] Recently it was shownthat solid lipid doxorubicin loaded nanoparticles have poten-tial to serve as a useful therapeutic approach to overcomethe chemoresistance of Adriamycin-resistant breast cancerKoren et al [28] Koshkaryev et al [29] and Etzerodt et al[30] showed that the entrapment of some chemical com-pounds inside modified liposomes (resp by antibody [28 30]and transferring [29]) causes an increase of apoptosis of can-cer cells Moreover the solid lipid nanoparticle system couldbe generally applied for the delivery ofmany chemotherapeu-tic agents in chemotherapy-resistant cancers [31]

Micelles are self-assemblies of amphiphiles that formsupramolecular core-shell structures in the aqueous environ-ment Hydrophobic interactions are the predominant drivingforce in the assembly of the amphiphiles in the aqueousmediumwhen their concentrations exceed the criticalmicelleconcentration Phospholipid Pluronic poly(L-amino acid)and polyester micelles are most often applied In [32] authorssummarize advances related to targeted anticancer drugdelivery to tumor sites using PMs via active and passivemechanisms (see below) PMs can be conjugated with diverseligands such as antibodies fragments epidermal growthfactors 120572-2-glycoprotein transferrin and folic acid

Over the past few decades there have been repeatedattempt to develop an ideal DDS that selectively acts againstdiseased cells but is not harmful to healthy cells PMs areone of the nanocarriers that can do this Also as mostof the anticancer drugs are poorly water-soluble PMs areconvenient drug carriers for carrying as well as targeting suchdrugs to tumors Collectively all these studies suggest thatdrugs encapsulated in micelles show enhanced therapeuticindex in solid tumors correlating to their passive targetingtaking advantage of tumor characteristics as well as activetargeting using various mechanisms and fewer side effectsin comparison with conventional drug formulations Amongthese a few PM formulations have been successfully devel-oped and a few more are at preclinical stage There is a direneed to translate these proven experimental advantageousconcepts into clinical practice to diminish the death rate fromcancers and increase hope in cancer chemotherapy [32]

Besidesmicelles there is the other groupof nanomaterialsforming self-assembling structure known as cell-penetrating


(a) (b)

(c) (d) (e) (f)

Figure 3 Schematic representation of nanocarriers (a) liposome (b)micelle (c) polymeric nanoparticle (d) dendrimer (e) carbonnanotubeand (f) carbon nanohorn

peptides (CPPs) These peptides could be applied to identifyhydrophobic anticancer drugs and intracellular delivery ofbiomolecules such as nucleic acids (siRNA pDNA) Theapplication of CPPs in novel delivery systems is favorablebecause of numerous advantages biocompatibility low tox-icity easy preparation and stability of the structure [33ndash36]Arukuusk et al [33] proved that the addition of hydrophobicmoieties to CPPs improves their properties during applica-tion as the nucleic acid delivery systems Deshayes et al [37]and Hou et al [38] described the creation of siRNA deliverysystem based on the CPPs The process of peptides self-assembly proceeds spontaneously during the contact withsiRNA A stable structure was formed mainly due to bothelectrostatic and hydrophobic interactions Those complexeseasily penetrate the cell and could be applied to successfullyprimary cell lines

Polymeric Nanoparticles Many methods have been devel-oped for preparing polymeric nanoparticles These methodscan be classified into two main categories according towhether the formulation requires a polymerization reactionor is achieved directly from a macromolecule or preformed

polymer Polymerization methods can be further classifiedinto emulsion and interfacial polymerization and there aretwo types of emulsion polymerization organic and aqueousdepending on the continuous phase Nanoparticles can bealso prepared directly from preformed synthetic or naturalpolymers and by desolvation of macromolecules Recentlythese polymeric systems have been prepared by nebulizationtechniques In [39] authors present all these methods includ-ing their detailed procedures and technological advantagesas well as providing several examples of encapsulants that areentrapped into or adsorbed to these particles The evolutionof nanoparticle preparation methods has been marked bythree aspects need for less toxic reagents simplification ofthe procedure to allow economic scale-up and optimizationto improve yield and entrapment efficiency Efficient drugentrapment and transition to large scale are of highestimportance to industrial applicability Depending on thephysicochemical characteristics of a drug it is now possibleto choose the best method of preparation and the bestpolymer to achieve an efficient entrapment of the drugNevertheless there are several problems remaining to besolvedThe process is not suitable to all drugs In addition the


postpreparative steps such as purification and preservationparticularly important for nanocapsules and residual solventanalysis must be extensively investigated Other difficultiessuch as the formation of an incomplete or of discontinuousfilm with inadequate stability of certain active componentsno reproducible or predictable release characteristics causesthat the final product is economically unfeasible [39] In [40]authors discuss possibilities of the polymeric nanoparticle-based technique of targeted drug delivery through the blood-brain barrier The biodistribution of novel nanoparticlesshowed two orders of magnitude greater efficiency in com-parison to other known drug carriers [40]

Dendrimers The role of dendrimers in delivery of differ-ent compounds (eg 5-fluorouracil primaquine phosphatedoxorubicin artemether tamsulosin indomethacin tropi-camide and pilocarpine) is presented in [41] Authors discussthe methods of intravenous transdermal ophthalmic andoral delivery There are different results that prove theversatility of dendrimers and some very important in vitrostudies with in vivo potential further endorse this versatilityMore detailed studies on the routes already investigated andstudies on other routes for dendrimer-mediated drug deliveryare required yet the existing data emphasized the potentialof dendrimers as drug carriers via various routes Howeverthe toxicological status of candidate dendrimers must beestablished conclusively before drawing any final conclusionsin this regard [41] Interesting review describing dendrimersis presented by Wen et al [42]

Carbon Nanotubes Basic properties and application of car-bon nanotubes in drug delivery were presented in [43ndash47]Different studies [48ndash54] describe the application of carbonnanotubes in DDS Firme III and Bandaru [45] describe themost popular strategies applied to increase the solubility ofnanotubesThe role of defects is also discussed and finally it isconcluded that the lack of centralized toxicity database limitsmakes the comparison between research results impossibleJain et al [55] present a novel cascade of chemical func-tionalization of multiwalled carbon nanotubes (MWCNTs)through chemical modification by a carbohydrate as D-galactose Galactose-conjugated MWCNTs were synthesizedinvolving the sequential steps of carboxylation acylationamine modification and finally galactose conjugation Themodification of MWCNTs with galactose was investigated bydifferent methods at every sequential step of functionaliza-tion Size and surface characteristics of chemically modifiedMWCNTs were monitored That galactosylation improveddispersibility ofMWCNTs in aqueous solventswas confirmedby investigation of their dispersion characteristics at differentpH values Thus the galactosylated MWCNTs could be usedfor delivery of different bioactive(s) as well as active ligand(galactose) based targeting to hepatic tissue [55]

Carbon nanohorns belong to a new class of carbonmaterials similar to carbon nanotubes Single-walled carbonnanohorn (SWNH) aggregates composed of thousands ofgraphitic tubules (similar in structure to single-walled CNTs)having wide diameters of 2ndash5 nm have a spherical struc-ture with a diameter of 50ndash100 nm On the basis of their

morphology they were classified into dahlia bud and seedtypes SWNHs contain no metal catalyst because they areproduced by laser ablation of a pure graphite target Thismeans that the effects of metal impurities can be excludedwhen determining toxic responses enabling investigation ofthe pure toxicological effects of nanometer-sized graphiticstructures To avoid potential health hazards caused by occu-pational exposure to SWNHs and to promote industrial andbiomedical applications of SWNHs the toxicity of SWNHsshould be proactively investigated from various aspectsComprehensively investigated in vivo and in vitro toxicities ofas-grown SWNHs lead to conclusions that carbon nanohornsare nontoxic [56]

There are much more important materials that can beused as drug carriers quantum dots [57 58] Pluronic [5960] mesoporous silica [61 62] nanoemulsions [63 64]drug nanocrystals [65 66] ceramic-based nanoparticles [67]albumin nanoparticles [68 69] nanogels [70 71] magneticnanoparticles [72 73] and so forth however in this reviewwe focus on (in our opinion) the most important onesAlso proteins are promising delivery agents They couldbe bioconjugated with drugs as albumin-bound paclitaxelforming Abraxane nanoparticle The formation of albumin-paclitaxel linking is prepared via homogenization processat high pressure [74] Abraxane is used as Cremophor EL-free formulation Thanks to this the system is more effectiveand less toxic than conventional drugs Chemotherapeuticagent is released fromAbraxane nanoparticle via the albuminreceptor in tumor blood vessel [74 75] Albumin-paclitaxelconjugationwas successfully applied against gastric [75] lung[74] and metastatic breast [76] cancer

4 Covalent Bond Formation between TargetedLigand and Nanocarrier

Various methods have been employed to link ligands withreactive groups of the surface of the nanocarriers and themethods can be classified into covalent and noncovalentconjugations [77] Common covalent coupling among theother methods involve conjugation of

(i) 2 thiol groups(ii) 2 primary amines(iii) a carboxylic acid and primary amine(iv) maleimide and thiol(v) hydrazide and aldehyde(vi) a primary amine and aldehyde

On the other hand the noncovalent bonding proceedingby physical association of targeted ligands to the nanocarriersurface has the advantage due to avoiding of rigorousdestructive reaction agents However there are some prob-lems such as weak bonding and low control of reaction Alsothe ligands may not be in the desired orientation after thedecoration process [9]

41 Amide Group The formation of amide bond proceedsby two stages During the first one carboxylic acid groups


O OO

ONH

N

NEDC

OH

(CH2)3 NH2HN

Scheme 1

on the carrier surface are activated by EDC 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide This reagent is usuallyapplied as carbodiimide which can form different chemicalstructures [77] EDC has good solubility in water This prop-erty enables direct application of EDC in aqueous solutionswithout addition of any organic compounds These condi-tions are suitable for the attachment of bioactive moleculesto the carrier surface During the reaction of EDC withcarboxylic group the active form of intermediate product O-acylisourea ester is formed The latter reacts with primaryamine forming amide bond (Scheme 1)

The main advantage of this method is that applied liganddoes not need any preliminary modifications usually causingthe loss of its activity [78 79] The possibility of activationof carboxylic groups derived from peptide could be someinconvenience in the case of amino acids These carboxylicgroups should have been locked for example by NHS (N-hydroxysuccinimide) Ishida et al [80] presented the additionof transferrin to liposome decorated by polyethylene glycolThey employed DSPE-PEG-COOH system which was thesource of required carboxylic acid groups The product inthe form of TF-PEG-liposome has interesting propertieslong time of biodistribution and large accumulation inbrain tumor Also Maruyama [81 82] or Blume [83] et aldescribed the formation of amide bond between ligand andliposome surface decorated with PEG-COOH Zengrsquos group[84] reported the attachment of EGFwith carboxylic group ofmicelle The product was employed in the targeted deliveryof drug Also the formation of amide bond formed on thecarbon nanotube surface was reported Ou et al [85] reportedthe SWCNTs modification by PEG and antibody (mAb)which was selectively encapsulated by integrin receptor ofcancer cellsThis process is schematically shown in Scheme 2

The SWNT-PEG-mAb is characterized by a high stabilitylow toxicity and high dispersion in water environment Theproduct was effectively captured by receptors of cancer cells[85] Zhang et al [86] modified oxidized MWCNT surfaceby biocompatible polyamidoamine dendrimer (PAMAM)The PAMAM chains were successfully conjugated into thesurface of carbon nanotubes This was confirmed by TEMimages The MWCNTs-PAMAM has very good dispersionand stability in aqueous solution Some extra tests performedby authors confirmed the efficiency of application in genetherapy [86] Dvir et al [87] showed the amide conjugationbetween targeted ligand and liposome This system during invitro tests was effectively used against cardiac cells [87] Chouet al [88] described the amide conjugation between anti-body and functionalized multiwalled carbon nanotubesThissystem was applied successfully in targeted photothermaltherapy Authors confirmed the better photothermal effect

of the f-MWCNTs connected with antibody for cancer celldestruction

The above described information was connected with theactivation of the carboxylic group on the surface of nanocar-rier Alternative method is associated with the activation ofprimary amine of nanocarrier as shown in Scheme 3

This process takes place when homobifunctional dithio-bis(succinimidyl propionate) (DSP) is used [77]TheDSPwasused in the synthesis of drug carrier which was applied in thetargeted breast cancer therapy [89] The DSP activates aminegroups of carriers Additionally application of NHS allowsthe formation of ester during the reaction with monoclonalantibody (trastuzumab) The methodology was very efficientand selective throughout cancer gene therapy

42 Thioether Group Thioether bond is formed during thereaction between thiol group and C

1carbon of maleimide

which is attached to the R2

carrier (see Scheme 4) Thereaction runs quickly and undermild conditions at the roomtemperature and in aqueous solution [12] Formed thioetherbond is stable within 24 hours in human serum even in thepresence of reduction agent for example DTT [77 78 90]Unfortunately the selectivity towards the thioether group for-mation is quite low in aqueous solution due to the side reac-tions such as intermolecular rearrangement or formationof disulfides The efficiency of reaction could be increasedthrough addition of activating agents for example SPDPN-succinimidyl-3-(2 pyridyldithio) propionate or SATA N-succinimidyl-S-acetylthioacetate Application of the systemscontaining the thioether bonds guarantees the high selectivityof delivery and the long time of distribution [12 78]

Kirpotin et al [91] described the synthesis of a selec-tive DDS based on liposome cholesterol and PEG modi-fied DSPE Antibodies were conjugated to the nanocarrierthrough free thiol group The synthesis is based on two ideasFirst is the coupling of antibody with double layer of lipo-some Alternatively the second provides a simple conjugationof ligand to the distal end of PEG chains Authors demon-strated that the uptake of anti-HER2-immunoliposome cor-related with density of cancer cell surface and with the effectof tumor reduction The efficiency of the process has beenincreasing with increasing amount of encapsulated antibodyFab Similar techniques with the use of different antibodieswere applied successfully by for example Maruyama et al[81] Allen et al [92] Hansen et al [93] Zalipsky et al [94]Park et al [95] and Ren et al [96] Produced systems wereeffectively used in drug delivery especially for doxorubicin

Anhorn et al [97] were pioneers in the synthesis ofDDS based on monoclonal antibody trastuzumab andnanoparticle doped with doxorubicin The conjugation hada nature of thioether bond The covalent bonding took placebetween thiol group of ligand and maleimide fragment ofa carrier (Scheme 5) Initially the carrier was modified bymolecules of doxorubicin Nanoparticles were activatedthrough poly(ethylene glycol)-R-maleimide-120596-NHS esterApplied ligand enabled effective uptake of anticancersystem by HER2 receptor of breast cancer cells The drug ischaracterized by a long time of circulation in blood without


HO

OO O O

O

OO

O

O

O

OO

O

O

O

O

OO

O O

O

O HN

O

O

O

HO

S

O NH NH

NH

OH

P

H

O O

O

O

N

S

OH

O

O

O

OH

N

OH

O O

O

P

P

O

O

(OCH2CH2)45

(OCH2CH2)45

(OCH2CH2)45

NH

NH

HO

+ EDCNHS

Protein A

Protein A

NH2

NH

NH2 +

Protein AOminus

Ominus

Ominus

Scheme 2

any side effects Nanoparticles had higher loading capacityof doxorubicin comparing with liposomes

The bioconjugation of protein to nanocarrier by thioetherbond was described by Gindyrsquos group [98] The carrierEG-b-PCL was modified by maleimide The BSA was usedas targeted ligand containing SH fragments The processoccurred in aqueous solution The covalent bioconjugation

between albumin and nanocarrier was confirmed by theanalysis of the nanocarrier volume before and after the pro-cess Higher concentration of protein in solution during thereaction caused increase in the number of BSA-nanoparticlebonds The targeted nanoparticles modified by thioetherbond between ligands that is aptamers and nanocarrierwere shown by Farokhzad et al [99] and Xiao et al [100]


DSPNH

O

SS

O

O

NO O

NHS

NH

OS

S

O

NH

NH2

NH2

Scheme 3

N

O

O

N

O

OS

SH +

R1

R1 R2R2C1C1

Scheme 4

The nanoparticle-aptamers system was successfully appliedin anticancer therapy The therapeutic effect was enhancedAlso Lu et al [101] successfully applied thioether boundbetween folic acid and functionalized carbon nanotubesThis new system was characterized in special properties tothe application in targeted delivery of DOX against cancertreatment

Alternative way is the coupling of thiol groups of thenanocarrier with maleimide fragments from the ligand Thethiol groups could be activated via amines or carboxylic acidgroups (Scheme 6)

The amide and thioether are the main bond types used toform the ligand-nanocarrier systems Other connection typesare only the derivatives [12]

43 Disulfide Group The disulfide bond is formed by theconjugation of two thiol groups (Scheme 7) The first grouporiginates from a nanocarrier while the other from ligand

The reduction of disulfide functionalities [102] or theapplication of suitable agents as SATAor SPDP [103] can formthiol groups of a ligand In order to form the thiol groups onnanocarrier surface one shouldmodify it with PDP-PE PDP-SA and PDP-PEG-DSPE where PDP fragment is a source ofthiol groups [12]

In [104] the formation of disulfide bond between lipo-somes and monoclonal antibody anti-My9 was describedThe carrier surface was decorated with PDP-SA The ligandwas modified with SPDP and kept its own immunoreactivelevel after the modification The obtained system in this wayacts strongly against human HL-60 promyelocytic leukemiacells [104] The formation of disulfide groups between tar-geted ligands (nanobodies) andmicelles was shown by Talelliet al [105]

44 Acetyl-Hydrazone Group The conjugation of hydrazidegroups to the nanocarrier surface occurs with aldehyde

groups of the ligands (Scheme 8) [77 78] As generally ligandsdo not possess the aldehyde structure the latter must beformed via oxidation of hydroxyl groups The oxidativeagents are usually sodium periodate [106] and galactoseoxidase [107] The major advantage of this method is therigorous control of the ligand modification [108] Howeverthe yield of the process in this technique is very poor [93]

Harding et al [109] described the linking of antibodiesC225 with liposome through acetyl-hydrazone group forma-tion The in vivo study showed that the activity of antibody inproduced immunoliposome was fully kept The system wascharacterized by long time of distribution in blood and highimmunological level Using this methodology one can easilycontrol immunoliposome-structure synthesis

45 Polycyclic Group TheDiels-Alder reaction (Scheme 9) isthe cycloaddition reaction between a diene and a dienophileAs a result bicyclic compound is formed The coupling ofligands and nanocarriers is beneficial because of high yield(close to 100) and easy synthesis under mild conditions[77]

The system of ligand-nanocarrier formed in the DAreaction causes specific bonds creation between ligand andcancer cells [12] Shi et al [110] used the DA reaction toconjugate antibody anti-HER2 with the polymeric nanocar-rier (Scheme 10) The carrier was synthesized via a softtemplating method The furan group on the external surfaceof carrier was the diene while maleimide group of antibodywas the dienophile Authors demonstrated the ability of thistechnique in creation of bioactive immunonanoparticles andconcluded that the versatility of the nanoparticle system canbe extended to create multiple functional delivery vehiclesFormed immunosystem was tested against breast cancer [111112]

46 The ldquoClick Chemistryrdquo The term ldquoclick chemistryrdquo (CC)was developed by Kolbrsquos group [113] and describes thecoupling of small elements and heteroatoms in the form ofR-X-R Reactions belonging to ldquoclick chemistryrdquo group arecharacterized by high efficiency stereospecificity and harm-less side products These processes need (i) mild conditions(ii) readily available reagents and (iii) easily removed solventfor example water [77]

Formed product should be stable under physiologicalconditions and should be easily isolated The main methodof the purification of final product is crystallization ordistillation [113] Within the CC one can find four majortypes [113 114]

(i) Cycloadditions for example Huisgen catalyticcycloaddition (Scheme 11)

(ii) nucleophilic substitution chemistry for example ringopening of heterocyclic electrophiles (Scheme 12)

(iii) carbonyl chemistry of the ldquononaldolrdquo type for exam-ple formation of ureas thioureas and hydrazones(Schemes 13-14)


n

n

NN

N

O

O

O

O

O

OO

O

O

OSH

S

H

NH

N

HOOC

HOOC

COOH

COOH

H

NH

NH2+Clminus

NH2+Clminus

NH2

NH2

NH2

NH2

Scheme 5

NO O

NH

O

SH NH

O

S

N

OO

OH

O

EDC DTT SPDP D

TTNH2

Scheme 6

S SR1R1 R2R2SH + HS

Scheme 7

NH NH N

O

O

OH

NH2

Scheme 8

(iv) additions to carbon-carbonmultiple bonds for exam-ple epoxidation and dihydroxylation (Schemes 15-16)

The major type of ldquoclick chemistryrdquo reactions is theHuisgen cycloaddition This process creates 123-triazole bythe 13-dipolar cycloaddition between azides and terminalalkynes in the presence of the catalyst Cu (I) [12 77 115]The most probable mechanism of the reaction is shown inScheme 17 [114]

Huisgenrsquos cycloaddition requires reagents tolerating aque-ous solution with broad range of pH level and biologicalmolecules [114] The effective ligand-liposome conjugation

O

O

N

O

ON

O

O

Scheme 9

based on Huisgenrsquos cycloaddition was described by Hassaneet al [116]The ligandwas120572-D-mannose derivativewith azidegroup The liposome possessed groups with triple bondsReported studies confirmed the essential role of addition ofCu (I) in order to increase efficiencyThe synthesized productwas almost perfect selective immunosystem [116]

The De et al [117] results confirmed the possibility ofldquoclick chemistryrdquo application in targeted anticancer therapyin order to conjugate the folic acid to themicellar nanocarrierAlso Lu et al [118] decorated nanoparticles by selectiveligands using Huisgenrsquos cycloaddition The process was effec-tive and almost 400 peptide molecules modified coumarinwere attached to each nanoparticle Fluorescence intensity of


H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

coumarin confirmed the efficiency of the reaction betweenpeptide connecting alkyne group and nanoparticle with azidegroup [118] Carbon nanotubes can also be modified byHuisgenrsquos cycloaddition Carbonnanotubes functionalized byHuisgenrsquos cycloadditionwere used in delivery ofmethotrexateto breast cancer cells [18]

We described here only themost popular covalent ligand-nanocarrier bonds which are often used in practice How-ever up to date much more types of covalent bonds are

known The other covalent link carbamate bond betweentargeted ligand and liposomes was shown by Sawant andTorchilin [119] An alternative and effective functionalizationmethod is the formation of amide bonds between azides andtriphosphine known as Staudingerrsquos reaction Generally itoccurs at room temperature and in aqueous solution anddoes not require any catalyst Zhang et al [120] attachedglycoliposome with lipid containing triphosphine fragmentsby Staudingerrsquos reaction This method is applicable to watersoluble molecules

Another type of covalent bond is the immobilization ofprimary amine with free aldehyde group of ligand by for-mation of Schiff rsquos base [78] This technique was used duringthe linking of the transferrin (TF) with poly(lactic acid)surrounded by cholesterol chains containing indomethacinmolecules The TF-nanocarrier was studied against gliomacell Experiment confirmed the bioactivity of transferring[121]

The main covalent interactions and essential conditionsof the processes are shown in Table 1

5 Physical and Chemical Bonding of Drug toNanocarrier Surface

Drugs applied in targeted anticancer therapy could be accu-mulated outside andor inside of nanocarriers (Figure 4)Thecouplings can be nonspecific (eg adsorption) or covalent[122]

The entrapment of a drug to nanocarrier is possibleduring the synthesis of nanocarrier or after the carrierformation The loading of chemotherapeutic agent dependson its solubility in the nanoparticle matrix the mass of themolecule the interaction of drug nanocarrier and the typeof functional groups present on a nanocarrier surface [123]A nanocarrier should be chemically resistant and shouldpossess high purity These properties are essential to controldelivery of drug [124]

Nanocarriers are often modified before the entrapmentof drugs The most popular method of functionalizationemploys polymers The medical practice uses both natu-ral and synthetic polymers for example polyethylene gly-col poly(lactic acid) N-(2-hydroxypropyl)methacrylamidecopolymer poly(L-glutamic acid) and poly(DL-lactic-co-glycolic acid) [124]The bioconjugation of drug with polymerleads to the extension of chemotherapeutic circulation andcontrolled drug release in targeted place [125ndash129] PEG hashigh solubility in water Moreover it is a good ldquocandidaterdquo forapplication inmedicine because of its biochemical propertiesbiodegradation minimal toxicity and controlled mechanicalproperties [125 130] Generally it is important to apply thepolymeric phase because it improves the drug accumulationin targeted place and generates the barrier between nanocar-rier and cancer cell [126 129]

51 Drug on the Internal Surfaces Nanocarriers The internalwalls of a nanocarrier (eg of CNT) in contrast to the externalwalls reveal high interaction energywith adsorbedmoleculesThe entrapped drug molecules inside the nanocarrier are


Table 1 Types of covalent ligand-carrier bindings

Name Functional group ofnanocarrier

Functional group ofligand Conditions of process Literature

Amide group Carboxylic Primary amine EDC NHS pH 75ndash852ndash24 h 4∘C lub RT [98 102ndash108]

Thioether group Maleimide Thiol pH gt 7 4ndash24 h RT [102 111ndash117]Acetyl-hydrazone group Hydrazide Hydroxylic 24 h 5-6∘C [124]Disulfide group Thiol Thiol pH 80 2ndash24 h 4∘C or RT [120]Diels-Alder Furan Maleimide pH 55 2ndash6 h 37∘C [125 126]ldquoClick chemistryrdquo (HDC) Azide Alkyne Cu (I) RT 2-3 h [131ndash134]Staudinger Azide Triphosphine PBS pH 74 6 h RT [135]Schiff rsquos base Primary amine Aldehyde pH 92 RT [138]

XHX

NuNu

Nucleophilic substitution chemistry

H3O+

X = O NR +SR +NR2

∙∙

Scheme 12

ldquoNonaldolrdquo carbonyl chemistry

R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14

isolated from external environment and they are protectedagainst early activation and degradation process Thus theinteraction of chemotherapeutic with healthy tissue is impos-sible [131 132]

Drug molecules can be entrapped by nanoextractionandor nanocondensation process (Figure 5) Nanoextrac-tion process occurs in liquid phase and at the room tempera-ture The idea is based on the selection of solvent adequateto the guest molecule Entrapped drugs should have pooraffinity to the solvent and high affinity to the nanocarrierWeak solubility of guest molecules in the solvent enablesformation of the suspension in liquid phase and the gradualdiffusion inside the nanocarrier [133]

Nanocondensation similarly as nanoextraction processis performed in liquid phase also at the room temperature

Ligand nanocarrier

Drug conjugation

Drug entrapment

Figure 4 The drug conjugation to surface of a carrier and anentrapment process (the figure is based on [122])

This process is relatively easy and quick Guest moleculesshould demonstrate high affinity to the solvent and to thenanocarrier Generally nanocondensation process is basedon the adsorption of solvent molecules on the externaland internal walls of nanocarriers Guest molecules migratethrough the thin layer of a solvent and are adsorbed in themost active centers via the van der Waals interactions [133]

Nanoextraction process was utilized by Ren and Pastorin[19] Authors loaded the drug hexamethylmelamine inside


R1

R1

R2

R2

[X] X+ Byproduct X = O NR +SR +NR2

Carbon multiple bond addition

Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17

CNTs C60-fullerenes were used as caps of open CNTs endsIn the first step CNTs were treated by a mixture of acids(HNO

3 H2SO4) During this process CNTs were opened

and functional groups (mainly carboxylic) were generatedCarboxylic fragments created the barrier for other moleculescompared to the drug ones Authors proved that suitablyfunctionalized CNTs are good candidates for drug nanocar-riers The stable structures of CNTs and C60 protected drugmolecules from circulation in blood before theywere releasedat specific places [19]

Studies concerning the cisplatin loaded inside SWCNTswere reported by Tripisciano et al [134] The system ofSWCNTs-cisplatin was used against colon cancer Authors

utilized a mixture of HNO3and H

2SO4acids in order to

open SWCNTs The drug was loaded inside SWCNTs bynanocondensation process Carbon material was dispersedin dimethylformamide (DMF) containing cisplatin (DDP)Carbon nanotubes limited the drug precipitation to aqueoussolution and its degradation Analysis of FTIR Raman spec-troscopy HR-TEM and EDX results confirmed the presenceof cisplatin inside SWCNTs structure Authors observed theproportional correlation between the anticancer propertiesand the amount of SWCNTs-DDPThe newDDS caused sim-ilar effect on the cancer cells as free drug However the novelsystem minimized the presence of side effects Another workof Tripisciano et al [135] reported the nanocondensation


Table 2 Effectiveness of DDP delivery by SWCNTs [134] and MWCNTs [135]

SWCNTs MWCNTsThe amount of DDP inside CNTs 21 120583g100 120583g of CNTs 136120583g100 120583g of CNTsThe amount of released DDP 68 95The rate of DDP release Slower FasterThe maximum release of DDP 72 h 48 h

SWCNT

Nanoextraction Ethanol

Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene

Figure 5 Scheme of the nanoextraction and nanocondensation processes (the figure is based on [133])

process used to the loading of cisplatin inside the MWCNTsThe method of entrapment process was similar to SWCNTsTable 2 summarizes the results for SWCNTs and MWCNTs

It is interesting that almost all drug molecules werereleased from MWCNTs This confirms the fact that carbonnanotubes can be successfully used as drug containers andtherefore they could serve as DDS Moreover according tothe authors inside DDP-MWCNT systems the specific inter-action did not occur in contrast to DDP-SWCNTs Thoseinteractions are probably responsible for slower and smallerpercentage release of DDP from SWCNTs Likewise Li et al[136] and Sui et al [137] described the encapsulated DDPinside carbon nanotubes Authors confirmed the improvedefficiency of anticancer therapy

Another interesting approach is the entrapment of DDPinside the nanoparticle covered with copolymer Grypariset al [138] used this container against colon cancer The

drug kept its activity after loading inside the nanocarrier Itwas delivered selectively to cancer cell The therapeutic effectwas better comparing with the traditional chemotherapyThe DDP-nanoparticle system was safe for human organismCisplatin can be also adsorbed within PMs [139 140] orliposomes [141 142] or carbon nanohorns [143 144] Thein vivo and in vitro studies were aimed against lymphoma(J6456) [141] colon (C26) [141 142] lunge [142ndash144] andstomach (MKN 45) [140] cancer cell Analysis confirmed thatthe DDP internalization inside nanocarriers improved thedrug activity and minimized side effects [139ndash142]

Hampel et al [145] used the entrapment process of carbo-platin inside carbon nanotubes They were opened and nextthe drug was placed inside the structure using wet impregna-tion method Authors confirmed the presence of carboplatininside the structure which protects the drug against the envi-ronment Carboplatin inhibited the growth of blood cancer


Drug

PLGA

Au layer

Figure 6 Schematic diagram of DOX-PLGA-Au (the is figure basedon [149])

cells The results confirm efficiency of carbon nanotubes aschemotherapeutic nanocarriers [145] In another interestingwork [146] an encapsulated Pt(IV) prodrug inside polymericnanoparticle was described Additionally the targeted ligandswere conjugated to the surface of nanoparticle This DDSinhibited growth of breast as well as prostate cancer cellsThis fact was connected with the protection of prodrugstructure by nanoparticles Moreover the biodistribution ofnovel system was better than that in the case for cisplatinMitra et al [147] reported the DDS based on doxorubicininside nanoparticle (diameter ca 100 nm) Drug was linkedwith additional remedy dextran The conjugation limitedside effects of doxorubicin The nanoparticle-DOX systemimproved therapeutic effect of drug Authors observed thereduction of tumor volume after 4 weeks Doxorubicin timecirculation in blood and amount inside cancer cell were betterin comparison to the traditional chemotherapy

Lince et al [148] and Park et al [149] described process ofdoxorubicin entrapment inside nanoparticle Lince et al [148]showednanoparticle based on biodegradable and biocompat-ible copolymerThe polymeric nanocarrier improved activityof drug which was placed inside and limited its potentialside effects Thus the comfort level and quality of patientrsquoslife increased Park et al [149] as well as Lince et al [148]described the encapsulation of doxorubicin inside polymericnanoparticle The polymer nanocarrier was covered withthe extra gold layer (Figure 6) The DOX-PLGA-Au systemcombined traditional chemotherapy and phototherapy Theefficiency was examined against cervical carcinoma Thenovel therapy reveals high therapeutic efficiency and shorttime of treatment The DOX-PLGA-Au system deliveredselectively drug to cancer cell and thus caused rise oftemperature which destroys cancer tissues The warmth wasgenerated as a result of absorption of NIR radiation by goldlayer

Doxorubicin in targeted anticancer therapy was usedsuccessfully within liposomes by Matsumura et al [150] orGabizon [151] and within micelles by Kataoka et al [152]Perche et al [153] Ebrahim Attia et al [154] or Cambon etal [155] Paclitaxel similarly as cisplatin or doxorubicin canbe adsorbed inside the nanocarrier Koziara et al [156] usedthis drug inside nanoparticle in vitro against brain cancercells U-118 and HCT-15 Authors proved the rise in cytotoxic

PEG outer shell

Hydrophobic inner core

PTXBlock copolymer

Figure 7 Schematic structure of NK105 (the is figure based on[160])

activity of drug and the limitation of the transmembranepump action Thus the uptake of a drug by cancer cellwas facilitated The new system penetrated through blood-brain barrier The stability of PTX was maintained duringthe entrapment process Ruan and Feng [157] proved thatthe release from nanoparticle is easy when the nanoparticleis covered with copolymer PLAndashPEGndashPLA Probably thiseffect occurs because hydrophilic fragments of PEG insidehydrophobic fragments of PLA increase the porosity [157]Fonseca et al [158] and Chan et al [159] also described theimmobilization of paclitaxel inside polymer nanoparticlesAuthors proved the efficiency of this method [158] Ham-aguchi et al [160] proposed the new form of PTX-NK105namely the drug inside micelle (Figure 7)

The delivery of NK105 to organism runs without Cre-mophor EL and ethanol in contrast to the traditional anti-cancer therapy The NK105 shows lower toxicity againstnervous system and better activity [160] Similarly Kim et al[161] and Wang et al [162] located paclitaxel within polymermicelle They confirmed the efficiency of novel form

52 Drug on the External Surfaces of Nanocarriers To con-struct the delivery system with a drug on external surfaceof a nanocarrier the chemical or physical conjugation isapplied This method is based on chemical properties andhigh surface area of a nanocarrier [132] Drug molecule islinked with functional groups of nanocarrier or polymercovering nanocarrier by covalent bonds forming ester amideor acetyl-hydrazone groupsThis type of coupling has usuallylow stability [163] and shows pH sensibility [164] Drugs withplane or aromatic structure can be adsorbed on the surface ofnanocarriers by 120587-120587 interactions [163]

521 Ester Group The ester bond formation between forexample doxorubicin and nanocarrier is possible as a resultof

(i) the bioconjugation of drugrsquos primary hydroxylicgroup (ndashC=OCH

2OH) with polymerrsquos carboxylic

group for example PLGA surrounding nanocarrier(Scheme 18)

(ii) the bioconjugation of drugrsquos primary amine groupwith polymerrsquos hydroxylic group (Scheme 19)


O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18

Yoo et al [127 128] described the formation of esterbondbetweenDOXandnanocarrier coveredwith PLGATheconjugation takes place between hydroxylic and carboxylicgroups [127] and between amine and hydroxylic groups [128]The novel systems were in vivo and in vitro tested againstcancer cell line HepG2 For both of the cases drug deliveredon the external surface was better absorbed by cancer cellthan the drug placed inside nanocarrier Authors showedthat the release of drug from nanocarrier is correlated withthe molecular mass of polymer placed on the surface ofnanocarrier [127]

Another anticancer drug as for example paclitaxel canbe conjugated to nanocarrier by ester bond Li et al [165]showed the bioconjugation of paclitaxel with polymericnanocarrier covered by poly(L-glutamic acid) (PG)The ther-apeutic activity of drug against OCa-1 cancer cells was higherthan during traditional chemotherapy Authors proved that

the PTX-PG conjugation has lower toxicity than free PTXThe novel system was better bonded to cancer cells andits circulation time was longer It was confirmed that labileester bond formation between polymeric nanocarrier andchemotherapeutic agent is useful method for the designingof targeted DDS Milas et al [166] used the same idea tolink paclitaxel with polymeric nanocarrier and confirmed theefficiency of the method

522 Acetyl-Hydrazone Group The hydrazone bond can beformed via the conjugation of carbonyl group with hydrazideone This nanocarrier-drug linking has high cytotoxicitylevel against selected cancer cell lines [167] Doxorubicinpossesses the carbonyl group which can combine withhydrazide group on the surface of nanocarrier coveringpolymer [168] The most popular polymers described in theliterature are poly(allyl glycidyl ether) [169] (Scheme 20)


C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19

N-(2-hydroxypropyl)methacrylamide copolymer [167] andpoly(ethylene oxide)-block-poly(allyl glycidyl ether) [168]

Vetvicka et al [169] proposedmicellar arrangement basedon amphiphilic diblock copolymer and doxorubicin Thissystem shows 20-time lower toxicity and longer time ofcirculation than the free drugThe therapeutic activity againstlymphoma EL-4 T was promising and 75 of mice popu-lation was cured completely and showed specific resistanceagainst new cancer cells [169]

Hruby et al [168] as well as Liu et al [170] describedthe formation of acetyl-hydrazone group between doxoru-bicin and nanocarrier covering poly(ethylene oxide)-block-poly(allyl glycidyl ether) [168] and poly(ethyleneimine)-polyethylene glycol [170] respectively Drug release was pH-sensitive it was faster in pH similar to that of endosomes (pH50) than in pH of blood plasma (pH 74) Probably this effectis related to the shift of equilibrium between free and bondedmolecules of doxorubicin to the direction of drug releasein lower pH [168] Yoo et al [171] showed the hydrazonebond between doxorubicin and polymericmicelles Drugwaslinked to the terminal hydrazide groups on the nanocar-rier poly(lactic acid) and methoxy-polyethylene glycol

Doxorubicin was delivered quickly to the cancer cell andpossessed high cytotoxicity [171]

Platinum drugs structural properties allow the formationof the hydrazone bond between drug and nanocarrier Aryalet al [172] described the bioconjugation of copolymer PEG-PLA with the hydrazide moieties PEG-PLA-NH-NH

2with

levulinic acid modified with Pt(IV) cisplatin analogue Theamount of the associated drug molecules was controlledPolymeric nanoparticles increased the cytotoxicity propertiesagainst ovarian cancer cellsThe drug loss during nanocarriercirculation in the blood was low in natural pH [172]

The hydrazone bond is used for the bioconjugationof paclitaxel with dendrimeric nanocarrier It was claimedthat also polyamidoamine dendrimers (PAMAM) could betreated as nanocarrier of chemotherapeutic agents The coreis built up with alkyl-diamine with tertiary amine branchesAmine groups allow the easy functionalization of dendrimerwith the drug The modification of the nanocarrier surfaceby paclitaxel was used against ovarian cancer cells Thetargeted ligand (protein LXW7) was also attached to thenanocarrier The method was effective and eliminated cancercells The drug-nanocarrier bond was broken at low pH


O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20

level and the drug was released [173] Rodrigues et al [174]demonstrated the bioconjugation of paclitaxel with poly-meric nanocarrier polyethylene glycolThe reaction requiredthe preliminarymodification of chemotherapeutic agentThemaleimide derivatives of paclitaxel were formed by bondingthe maleimide fragments with drug molecule At the firststep the synthesis of drug ester derivative in position C-21015840-OHby using 4-acetylbenzoic acid occurredThe intermediateproduct is a donor of carboxylic group which was essential tothe attachment of maleimide derivative containing hydrazidefragment Authors proved the efficiency of the method Thestructure of drug was protected and its activity was improvedvia in vitro and in vivo studies

523 Amide Group Amide bond is formed during thereaction of carboxylic group with a nucleophile contain-ing the primary amine Feazell et al [175] described theformation of amide bonds between platinum compoundand SWCNTs Functionalized SWCNTs were covered byphospholipid containing amine groups The compound cct-[Pt(NH

3)2Cl2(OEt)(O

2CCH2CH2CO2H)] as prodrug was

bonded to the modified carbon nanotubes The novel systemwas taken up effectively by endosomes where pH level is lowand as a consequence cisplatin (DDP) which was releasedas active compound This method enhances weak circulationand distribution of drug in blood and minimizes the DDPtoxicity against healthy cells [175]

Liu et al [48] described the conjugations between pacli-taxel and SWCNTs by the amide bond The structure ofSWNCTs was functionalized by polyethylene glycol withamine fragment (PEG-NH

2) The carboxylic group was

introduced to the drug structure in C-21015840-OH position Drugshowed better solubility than clinical form of paclitaxelTaxol Additionally the SWCNTs-PTX system showed longertime distribution in the blood the 10-time larger uptake bycancer cells and stronger therapeutic effect in comparisonto Taxol The new form of paclitaxel minimized the tumor

volume at drug doses as small as 5mgkg The side effectswere lower than those during traditional therapy [48]

Doxorubicin was attached to the nanocarrier using theamide bond in [166 176] Lai et al [177] discussed the bio-conjugation of drug to the dendrimer The efficiency of novelsystemwas examined against gingival cancer cell line Ca9-22The cytotoxicity was improved comparing with free drug

524 Disulfide Group Disulfide bond between drug andnanocarrier is created in order to obtain the systems for selec-tive delivery and accumulation of and large amount of drugsMost of the drugs and nanocarriers do not possess thiolgroups in the structure and thus the method requests prelim-inary modificationThe chemotherapeutic structures possessusually amine and carboxylic groups and these groups maybe linked with peptides the source of the thiol groups Thismakes the formation of disulfide bond possible [164]

Paclitaxel bonded by disulfide coupling to SWCNTs wasused against leukemia line L1210FR The disulfide linkagewas attached in C-21015840 position of the drug SWCNTs weremodified by oxidation conjugation of amide fragments andfunctionalization by amine groups Biotin was used as thetargeted ligand The new DDS successfully destroyed cancercells The chemotherapeutic agent was released when thePTX-SWCNTs were entrapped inside cancer cells [178]

525 Adsorption Chemotherapeutic agents can be accumu-lated on the external surface of nanocarriers also by phys-ical adsorption (eg by electrostatic interactions betweennanocarrier surface and biomolecule) [179ndash190]

Kataoka et al [180] described physical adsorption of dox-orubicin on the surface of micellar nanocarrier (poly(ethy-lene glycol)-poly(b-benzyl-Laspartate)) (PEG-PBLA) Thedrug-nanocarrier systemwas formed by the 120587-120587 interactionsbetween anthracycline groups of DOX and benzyl residuesof PBLA segments Drug on the nanocarrier revealed greateractivity comparing with the free drug Micelles allowed


longer circulation time of doxorubicin in the blood Greishet al [181] proved 13-time higher drug concentration insidetumor S-180 when the doxorubicin was loaded on thesurface of polymeric micelle The new system showed bettertherapeutic properties and lower side effects than what isobserved in traditional therapy [180 181]

Another study [182] reported the noncovalent attachmentof doxorubicin to carbon nanotubes MWCNTs were coveredby triblock copolymer Pluronic F127 Authors proved the riseof DOX cytotoxicity against breast cancer cells line MCF7in comparison to standard therapy The use of MWCNTsas drug nanocarrier improved the activity of drug and itsuptake by cancer cells The efficiency of this MWCNTs-DOX complex was determined by effective release of drug insuitable time and place [182] Heister et al [183] presented anoncovalent complex of doxorubicin with single-walled car-bon nanotubes As targeted ligandmonoclonal antibody wasused Authors proved that SWCNTs can be applied in orderto improve the DDS High activity of DOX adsorbed on thesurface of oxidized carbon nanotubes ormodified by polycar-bohydrate PEG PEG44-PPS20 copolymer and poly(acrylicacid) was proved byWangrsquos [191] Zhangrsquos [184] Liursquos [185] DiCrescenzorsquos [186] and Lursquos [101] groups respectively

Paclitaxel can be adsorbed on the modified surface ofnanocarrier Tian et al [187] adsorbed PTX on carbonnanotubes by 120587-120587 interactionsMWCNTs were covered withpolyethyleneimine Next the reaction was performed withfolic acid as targeted ligandDrug showed the rise of solubilityand was selectively taken up by cancer cells The activity invitro of paclitaxel loaded on the surface of carbon nanotubeswas greater in comparison to free drug used during tradi-tional therapy [187]

Adsorption of platinum complex is determined by thestability of nitrogen ligand and the mobility of chloride ionThe positively charged platinum aqua-complex is stronglyadsorbed on nanocarriers Adsorption of cisplatin is deter-mined by electrostatic interactions [188] Barroug et al [189]studied cisplatin adsorption on nanocarrier and showed therise of drug adsorption and the rate of drug release withthe rise in temperature The cytotoxicity was tested in vitroagainst K8 clonal murine osteosarcoma cell line Electrostaticinteractions between cisplatin and the surface of nanoparticledid not change the drug activity Cisplatin analogue that iscarboplatin was adsorbed on the surface of carbonnanotubesby Arlt et al [190] Adsorbed drug stopped the growth oftumor and the efficiency of loaded chemotherapeutic agentwas greater than observed for free carboplatin

Bonding by electrostatic interactions as the method ofthe drug and nanocarrier bioconjugation is lees popularin comparison to covalent linking The covalent couplingsare preferred due to the possibility of controlling the drugamount and the orientation during synthesis [179]

We described only selected drug accumulation possi-bilities Ionic drug complexes for example cisplatin withnanoparticles based on hyaluronic acid [192] or doxorubicinwith ionic complex [193] are also discussed in the literatureThe covalent functionalization by 13-dipolar cycloadditionis also widely used for the bioconjugation of drugs andnanocarrier Terminal amine groups on the nanocarrier

surface are ideal and reactive centers for the coupling ofbiomolecules [45] Pastorin et al [194] discussed the func-tionalization of MWCNTs via 13-dipolar cycloadditionActive groups formed on the surface of carbon nanotubeswere attached with active carboxylic fragments of drugmethotrexate This coupling minimized the intracellularuptake barrier and caused the increase in the efficiency

Next interesting attempt is the attachment of two anti-cancer drugs to one nanocarrier Few nanocarriers (egpolymeric nanoparticle and liposomes) have characteristicproperties which are essential during the coupling of differentchemotherapeutic agents Drugs attached together should beeasily hydrolyzed because they are independently aggregatedinside cancer cells [20] This methodology minimizes thedrug resistance of cancer cells and releases essential drug dose[195]

Aryal et al [20] showed the accumulation of paclitaxeland gemcitabine hydrochloride on the nanoparticle surfaceThe cytotoxicity of drugs comparing with their free analogswas greater against human pancreatic cancer cells XPA3 [20]Similarly Zhang et al presented the linking of doxorubicinwith docetaxel [196]

6 Summary

Synthesis of a novel DDS by application of nanomedicinerecipes is very important in anticancer therapy Applicationof nanocarriers can improve the activity of drugs Nanocar-riers isolate drug molecules from biological environmentand consequently minimize the enzymatic degradation ofchemotherapeutic agent [125] Additionally nanocarrierscause the rise in solubility and prolong the time of distribu-tion in the blood Those novel systems break the biologicalbarriers for example blood brain As a consequence a drugis delivered into the targeted cells hardly available duringstandard chemotherapy The new DDS as targeted ligand-nanocarrier drug fulfills all requirements of the effectiveand safe anticancer therapy (i) adequate concentration (ii)effective dose and (iii) high cytotoxicity of chemotherapeuticagents [195] The delivery of drug by the application ofnanocarriers opens the way to the treatment of diseasesshowing so-called multidrug resistance [122] Drugs canbe aggregated on the external or internal surface of ananocarrier Suitable modification of nanocarrier facilitatesthe chemical bioconjugation of drug or targeted ligand viathe formation the functionalities as amide ester disulfide oracetyl-hydrazone groups The most important factor is thestructure of drug or ligand determining the type of reaction

The results of recent in vitro and in vivo studies [195ndash198] suggest that described novel DDS should be moreeffective and successful against cancer cells in comparisonto traditional chemotherapy Application of nanovehiclesin targeted active treatment seems to be very promisingdirection in current material science and medicine

Abbreviations

BSA Bovine serum albuminCC Click chemistry


CNT Carbon nanotubeDA Diels-Alder reactionDDP CisplatinDDS Drug delivery dystemDMAP NN-DimethylaminopyridineDMF DimethylformamideDOX DoxorubicinDSP Dithiobis(succinimidyl propionate)DSPE PhosphatidylethanolamineDTT DithiothreitolEDC 1-Ethyl-3-(3-dimethylaminopropyl)

carbodiimideEDX Energy-dispersive X-ray spectroscopyEGF Epidermal growth factorEPR Enhanced permeability and retention effectFmoc-Osu 9-Fluorenylmethoxycarbonyl-N-

hydroxysuccinimideFTIR Fourier transform infrared spectroscopyHR-TEM High-resolution transmission electron micro-

scopyMWCNT Multiwalled carbon nanotubeNHS N-HydroxysuccinimidePAGE Poly(allyl glycidyl ether)PAMAM Polyamidoamine dendrimerPCL Poly(120576-caprolactone)PBLA Poly(120573-benzyl-L-aspartate)PDP PyridyldithiopropionatePDP-PE N-[3-(2-Pyridyldithio)propionyl]phosphati-

dylethanolaminePDP-SA N-[3-(2-Pyridylthio)propionyl]-stearylaminePEG Polyethylene glycolPEG-b-PCL Poly(ethylene glycol)-b-poly(120576-caprolactone)PEO-PAGE Poly(ethylene oxide)-block-poly(allyl glycidyl

ether)PEG-PBLA Poly(ethylene glycol)-poly(b-benzyl-Laspar-

tate)PG poly(L-glutamic acid)PLA Poly(lactic acid)PLGA Poly(DL-lactic-co-glycolic acid)PM Polymeric micellePPS Poly(propylene sulfide)PTX PaclitaxelPyBroP Bromo-tris-pyrrolidino-phosphonium

hexafluorophosphateRT Room temperatureSATA N-Succinimidyl-S-acetylthioacetateSPDP N-Succinimidyl-3-(2

pyridyldithio)propionateSWCNT Single-walled carbon nanotubeSWNH Single-walled carbon nanohornTEA TriethylamineTEM Transmission electron microscopyTF Transferrin

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The paper was supported by NSC Grant DEC-201101BST501192

References

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[2] B Mishra B B Patel and S Tiwari ldquoColloidal nanocarriersa review on formulation technology types and applicationstoward targeted drug deliveryrdquo Nanomedicine NanotechnologyBiology and Medicine vol 6 no 1 pp 9ndash24 2010

[3] S K Sahoo S Parveen and J J Panda ldquoThe present andfuture of nanotechnology in human health carerdquoNanomedicineNanotechnology Biology and Medicine vol 3 no 1 pp 20ndash312007

[4] R A Freitas Jr ldquoWhat is nanomedicinerdquo NanomedicineNanotechnology Biology and Medicine vol 1 no 1 pp 2ndash92005

[5] K Kostarelos ldquoThe emergence of nanomedicine a field in themakingrdquo Nanomedicine vol 1 no 1 pp 1ndash3 2006

[6] CWang S Ravi G VMartinez et al ldquoDual-purpose magneticmicelles for MRI and gene deliveryrdquo Journal of ControlledRelease vol 163 no 1 pp 82ndash92 2012

[7] R GMendes A Bachmatiuk B Buchner G Cuniberti andMH Rummeli ldquoCarbon nanostructures as multi-functional drugdelivery platformsrdquo Journal of Materials Chemistry B vol 1 no4 pp 401ndash428 2013

[8] C Wei ldquoThe valuable and significant role of NanomedicinerdquoNanomedicine Nanotechnology Biology and Medicine vol 1no 4 p 285 2005

[9] O M Koo I Rubinstein and H Onyuksel ldquoRole of nanotech-nology in targeted drug delivery and imaging a concise reviewrdquoNanomedicine vol 1 no 3 pp 193ndash212 2005

[10] T Flynn and C Wei ldquoThe pathway to commercialization fornanomedicinerdquo Nanomedicine Nanotechnology Biology andMedicine vol 1 no 1 pp 47ndash51 2005

[11] N Bertrand J Wu X Xu N Kamaly and O C FarokhzadldquoCancer nanotechnology the impact of passive and activetargeting in the era of modern cancer biologyrdquo Advanced DrugDelivery Reviews vol 66 pp 2ndash25 2014

[12] Y Liu H Miyoshi and M Nakamura ldquoNanomedicine fordrug delivery and imaging a promising avenue for cancertherapy and diagnosis using targeted functional nanoparticlesrdquoInternational Journal of Cancer vol 120 no 12 pp 2527ndash25372007

[13] N K Mehra V Mishra and N K Jain ldquoA review of ligandtethered surface engineered carbon nanotubesrdquo Biomaterialsvol 35 no 4 pp 1267ndash1283 2014

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[19] Y Ren andG Pastorin ldquoIncorporation of hexamethylmelamineinside capped carbon nanotubesrdquo Advanced Materials vol 20no 11 pp 2031ndash2036 2008

[20] S Aryal C-M J Hu and L Zhang ldquoCombinatorial drugconjugation enables nanoparticle dual-drug deliveryrdquo Smallvol 6 no 13 pp 1442ndash1448 2010

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[30] A Etzerodt M B Maniecki J H Graversen H J Moller VP Torchilin and S K Moestrup ldquoEfficient intracellular drug-targeting of macrophages using stealth liposomes directed tothe hemoglobin scavenger receptor CD163rdquo Journal of Con-trolled Release vol 160 no 1 pp 72ndash80 2012

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[40] N Mody R K Tekade N K Mehra P Chopdey and NK Jain ldquoDendrimer liposomes carbon nanotubes and PLGAnanoparticles one platform assessment of drug delivery poten-tialrdquo AAPS PharmSciTech vol 15 no 2 pp 388ndash399 2014

[41] U Gupta H B Agashe A Asthana and N K JainldquoDendrimers novel polymeric nanoarchitectures for solubilityenhancementrdquo Nanomedicine vol 2 no 2 pp 66ndash73 2006

[42] S Wen H Liu H Cai M Shen and X Shi ldquoTargeted andpH-responsive delivery of doxorubicin to cancer cells usingmultifunctional dendrimer-modifiedmulti-walled carbon nan-otubesrdquo Advanced Healthcare Materials vol 2 no 9 pp 1267ndash1276 2013

[43] M Foldvari andM Bagonluri ldquoCarbon nanotubes as functionalexcipients for nanomedicines II Drug delivery and biocom-patibility issuesrdquo Nanomedicine Nanotechnology Biology andMedicine vol 4 no 3 pp 183ndash200 2008

[44] A Shahi ldquoCarbon nanotubes a noval carrier system for drugdelivery and cancer therapyrdquo International Journal of Pharmaand Bio Sciences vol 5 no 3 pp 298ndash306 2014

[45] C P Firme III and P R Bandaru ldquoToxicity issues in the applica-tion of carbon nanotubes to biological systemsrdquoNanomedicineNanotechnology Biology and Medicine vol 6 no 2 pp 245ndash256 2010

[46] J Y Hwang U S Shin W C Jang J K Hyun I B Walland H W Kim ldquoBiofunctionalized carbon nanotubes in neuralregeneration a mini-reviewrdquo Nanoscale vol 5 no 2 pp 487ndash497 2013

[47] H J Zhang C Chen LHou et al ldquoTargeting and hyperthermiaof doxorubicin by the delivery of single-walled carbon nan-otubes to EC-109 cellsrdquo Journal of Drug Targeting vol 21 no3 pp 312ndash319 2013

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[52] N W S Kam T C Jessop P A Wender and H DaildquoNanotube molecular transporters internalization of carbonnanotube-protein conjugates into mammalian cellsrdquo Journal ofthe American Chemical Society vol 126 no 22 pp 6850ndash68512004

[53] Z Liu S Tabakman K Welsher and H Dai ldquoCarbon nan-otubes in biology and medicine in vitro and in vivo detectionimaging and drug deliveryrdquoNano Research vol 2 no 2 pp 85ndash120 2009

[54] Z Liu S M Tabakman Z Chen and H Dai ldquoPreparationof carbon nanotube bioconjugates for biomedical applicationsrdquoNature Protocols vol 4 no 9 pp 1372ndash1382 2009

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[58] R Lehner X Wang S Marsch and P Hunziker ldquoIntelligentnanomaterials for medicine carrier platforms and targetingstrategies in the context of clinical applicationrdquo Nanomedicinevol 9 no 6 pp 742ndash757 2013

[59] H Hosseinzadeh F Atyabi R Dinarvand and S N OstadldquoChitosan-Pluronic nanoparticles as oral delivery of anticancergemcitabine preparation and in vitro studyrdquo InternationalJournal of Nanomedicine vol 7 pp 1851ndash1863 2012

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[65] E Che X Zheng C Sun D Chang T Jiang and S WangldquoDrug nanocrystals a state of the art formulation strategy forpreparing the poorly water-soluble drugsrdquo Asian Journal ofPharmaceutical Sciences vol 7 no 2 pp 85ndash95 2012

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[67] I Roy T Y Ohulchanskyy H E Pudavar et al ldquoCeramic-based nanoparticles entrapping water-insoluble photosensitiz-ing anticancer drugs a novel drug-carrier system for photody-namic therapyrdquo Journal of the American Chemical Society vol125 no 26 pp 7860ndash7865 2003

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[69] HWartlick B Spankuch-Schmitt K Strebhardt J Kreuter andK Langer ldquoTumour cell delivery of antisense oligonuclceotidesby human serum albumin nanoparticlesrdquo Journal of ControlledRelease vol 96 no 3 pp 483ndash495 2004

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modified with amphipathic poly(ethylene glycol) s conjugatedat their distal terminals to monoclonal antibodiesrdquo Biochimicaet Biophysica Acta vol 1234 no 1 pp 74ndash80 1995

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[83] G Blume G Cevc M D J A Crommelin I A J MBakker-Woudenberg C Kluft andG Storm ldquoSpecific targetingwith poly(ethylene glycol)-modified liposomes coupling ofhoming devices to the ends of the polymeric chains combineseffective target binding with long circulation timesrdquo Biochimicaet Biophysica ActamdashBiomembranes vol 1149 no 1 pp 180ndash1841993

[84] F Zeng H Lee and C Allen ldquoEpidermal growth factor-conjugated poly(ethylene glycol)-block- poly(120575-valerolactone)copolymermicelles for targeted delivery of chemotherapeuticsrdquoBioconjugate Chemistry vol 17 no 2 pp 399ndash409 2006

[85] Z Ou B Wu D Xing F Zhou H Wang and Y TangldquoFunctional single-walled carbon nanotubes based on an inte-grin 120572

1199071205733monoclonal antibody for highly efficient cancer cell

targetingrdquo Nanotechnology vol 20 no 10 Article ID 1051022009

[86] B Zhang Q Chen H Tang et al ldquoCharacterization of andbiomolecule immobilization on the biocompatiblemulti-walledcarbon nanotubes generated by functionalizationwith polyami-doamine dendrimersrdquo Colloids and Surfaces B Biointerfacesvol 80 no 1 pp 18ndash25 2010

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[94] S Zalipsky M Newman P Bhagya and C M Woodle ldquoModelligands linked to the polymer-chains on liposomal surfacesapplication of a new functionalized polyethylene glycol-lipidconjugaterdquoPolymericMaterials Science and Engineering vol 67pp 519ndash520 1993

[95] J W Park K Hong P Carter et al ldquoDevelopment of anti-p185HER2 immunoliposomes for cancer therapyrdquo Proceedingsof the National Academy of Sciences of the United States ofAmerica vol 92 no 5 pp 1327ndash1331 1995

[96] J Ren S Shen D Wang et al ldquoThe targeted delivery ofanticancer drugs to brain glioma by PEGylated oxidized multi-walled carbon nanotubes modified with angiopep-2rdquo Biomate-rials vol 33 no 11 pp 3324ndash3333 2012

[97] M G Anhorn S Wagner J Kreuter K Langer and H VonBriesen ldquoSpecific targeting of HER2 overexpressing breastcancer cells with doxorubicin-loaded trastuzumab-modifiedhuman serum albumin nanoparticlesrdquo Bioconjugate Chemistryvol 19 no 12 pp 2321ndash2331 2008

[98] M E Gindy S Ji T R Hoye A Z Panagiotopoulos andR K Prudrsquohomme ldquoPreparation of poly(ethylene glycol) pro-tected nanoparticles with variable bioconjugate ligand densityrdquoBiomacromolecules vol 9 no 10 pp 2705ndash2711 2008

[99] O C Farokhzad S Jon A Khademhosseini T-N T Tran DA LaVan and R Langer ldquoNanoparticle-aptamer bioconjugatesa new approach for targeting prostate cancer cellsrdquo CancerResearch vol 64 no 21 pp 7668ndash7672 2004

[100] Z Xiao E Levy-Nissenbaum F Alexis et al ldquoEngineering oftargeted nanoparticles for cancer therapy using internalizingaptamers isolated by cell-uptake selectionrdquo ACS Nano vol 61no 1 pp 696ndash704 2012

[101] Y J Lu K CWei C C MMa S Y Yang and J P Chen ldquoDualtargeted delivery of doxorubicin to cancer cells using folate-conjugated magnetic multi-walled carbon nanotubesrdquo Colloidsand Surfaces B Biointerfaces vol 89 no 1 pp 1ndash9 2012

[102] F J Martin W L Hubbell and D Papahadjopoulos ldquoImmun-ospecific targeting of liposomes to cells a novel and efficientmethod for covalent attachment of Fabrsquo fragments via disulfidebondsrdquo Biochemistry vol 20 no 14 pp 4229ndash4238 1981

[103] V O Ivanov S N Preobrazhensky V P Tsibulsky V R BabaevV S Repin and V N Smirnov ldquoLiposome uptake by culturedmacrophages mediated by modified low-density lipoproteinsrdquoBiochimica et Biophysica Acta vol 846 no 1 pp 76ndash84 1985

[104] M S Shaik N Kanikkannan and M Singh ldquoConjugation ofanti-My9 antibody to stealthmonensin liposomes and the effectof conjugated liposomes on the cytotoxicity of immunotoxinrdquoJournal of Controlled Release vol 76 no 3 pp 285ndash295 2001

[105] M Talelli S Oliveira C J F Rijcken et al ldquoIntrinsically activenanobody-modified polymeric micelles for tumor-targetedcombination therapyrdquo Biomaterials vol 34 no 4 pp 1255ndash1260 2013

[106] D E L deMenezes LM Pilarski and TM Allen ldquoIn vitro andin vivo targeting of immunoliposomal doxorubicin to humanB-cell lymphomardquo Cancer Research vol 58 no 15 pp 3320ndash33301998

[107] M M Chua S T Fan and F Karush ldquoAttachment ofimmunoglobulin to liposomal membrane via protein carbohy-draterdquo Biochimica et Biophysica Acta vol 800 no 3 pp 291ndash300 1984

[108] S M Chamow T P Kogan D H Peers R C Hastings R AByrn and A Ashkenazi ldquoConjugation of soluble CD4 withoutloss of biological activity via a novel carbohydrate-directedcross-linking reagentrdquo The Journal of Biological Chemistry vol267 no 22 pp 15916ndash15922 1992

[109] J A Harding C M Engbers M S Newman N I Gold-stein and S Zalipsky ldquoImmunogenicity and pharmacokineticattributes of poly( ethylene glycol)-grafted immunoliposomesrdquo


Biochimica et Biophysica ActamdashBiomembranes vol 1327 no 2pp 181ndash192 1997

[110] M Shi J H Wosnick K Ho A Keating and M S ShoichetldquoImmuno-polymeric nanoparticles by Diels-Alder chemistryrdquoAngewandte Chemie vol 46 no 32 pp 6126ndash6131 2007

[111] A D de Araujo J M Palomo J Cramer et al ldquoDiels-Alderligation and surface immobilization of proteinsrdquo AngewandteChemie International Edition vol 45 no 2 pp 296ndash301 2005

[112] V Marchan S Ortega D Pulido E Pedroso and A GrandasldquoDiels-Alder cycloadditions in water for the straightforwardpreparation of peptidendasholigonucleotide conjugatesrdquo NucleicAcids Research vol 34 no 3 p e24 2006

[113] H C Kolb M G Finn and K B Sharpless ldquoClick chemistrydiverse chemical function from a few good reactionsrdquo Ange-wandte Chemie International Edition vol 40 no 11 pp 2004ndash2021 2001

[114] C D Hein X-M Liu and D Wang ldquoClick chemistry apowerful tool for pharmaceutical sciencesrdquo PharmaceuticalResearch vol 25 no 10 pp 2216ndash2230 2008

[115] B Jeong Y H Bae D S Lee and S W Kim ldquoBiodegradableblock copolymers as injectable drug-delivery systemsrdquo Naturevol 388 no 6645 pp 860ndash862 1997

[116] F S Hassane B Frisch and F Schuber ldquoTargeted liposomesconvenient coupling of ligands to preformed vesicles usingldquoclick chemistryrdquordquo Bioconjugate Chemistry vol 17 no 3 pp849ndash854 2006

[117] P De S R Gondi and B S Sumerlin ldquoFolate-conjugated ther-moresponsive block copolymers highly efficient conjugationand solution self-assemblyrdquo Biomacromolecules vol 9 no 3 pp1064ndash1070 2008

[118] J Lu M Shi and M S Shoichet ldquoClick chemistry function-alized polymeric nanoparticles target corneal epithelial cellsthrough RGD-cell surface receptorsrdquo Bioconjugate Chemistryvol 20 no 1 pp 87ndash94 2009

[119] R R Sawant and V P Torchilin ldquoChallenges in development oftargeted liposomal therapeuticsrdquoThe AAPS Journal vol 14 no2 pp 303ndash315 2012

[120] H Zhang Y Ma and X-L Sun ldquoChemically-selective surfaceglyco-functionalization of liposomes through Staudinger liga-tionrdquo Chemical Communications no 21 pp 3032ndash3034 2009

[121] M-Q Gu X-B Yuan C-S Kang et al ldquoSurface biofunction-alization of PLA nanoparticles through amphiphilic polysac-charide coating and ligand coupling evaluation of biofunction-alization and drug releasing behaviorrdquo Carbohydrate Polymersvol 67 no 3 pp 417ndash426 2007

[122] D Peer J M Karp S Hong O C Farokhzad R Margalit andR Langer ldquoNanocarriers as an emerging platform for cancertherapyrdquo Nature Nanotechnology vol 2 no 12 pp 751ndash7602007

[123] B S Wong S L Yoong A Jagusiak et al ldquoCarbon nanotubesfor delivery of small molecule drugsrdquo Advanced Drug DeliveryReviews vol 65 no 15 pp 1964ndash2015 2013

[124] R Sinha G J Kim S Nie and D M Shin ldquoNanotechnologyin cancer therapeutics bioconjugated nanoparticles for drugdeliveryrdquoMolecular CancerTherapeutics vol 5 no 8 pp 1909ndash1917 2006

[125] V Torchilin ldquoMultifunctional and stimuli-sensitive pharma-ceutical nanocarriersrdquo European Journal of Pharmaceutics andBiopharmaceutics vol 71 no 3 pp 431ndash444 2009

[126] D R Khan ldquoThe use of nanocarriers for drug delivery in cancertherapyrdquo Journal of Cancer Science andTherapy vol 2 no 3 pp58ndash62 2010

[127] H S Yoo KH Lee J E Oh andTG Park ldquoIn vitro and in vivoanti-tumor activities of nanoparticles based on doxorubicin-PLGA conjugatesrdquo Journal of Controlled Release vol 68 no 3pp 419ndash431 2000

[128] H S Yoo J E Oh K H Lee and T G Park ldquoBiodegradablenanoparticles containing doxorubicin-PLGA conjugate for sus-tained releaserdquo Pharmaceutical Research vol 16 no 7 pp 1114ndash1118 1999

[129] T L Moore J E Pitzer R Podila et al ldquoMultifunctionalpolymer-coated carbon nanotubes for safe drug deliveryrdquo Parti-cle and Particle Systems Characterization vol 30 no 4 pp 365ndash373 2013

[130] L Juillerat-Jeanneret ldquoThe targeted delivery of cancer drugsacross the bloodndashbrain barrier chemicalmodifications of drugsor drug-nanoparticlesrdquo Drug Discovery Today vol 13 no 23-24 pp 1099ndash1106 2008

[131] S K Vashist D Zheng K Al-Rubeaan J H T Luong and F-S Sheu ldquoAdvances in carbon nanotube based electrochemicalsensors for bioanalytical applicationsrdquo Biotechnology Advancesvol 29 no 2 pp 169ndash188 2011

[132] R Marega and D Bonifazi ldquoFilling carbon nanotubes fornanobiotechnological applicationsrdquo New Journal of Chemistryvol 38 no 1 pp 22ndash27 2014

[133] M Yudasaka K Ajima K Suenaga T Ichihashi A Hashimotoand S Iijima ldquoNano-extraction and nano-condensation forC60

incorporation into single-wall carbon nanotubes in liquidphasesrdquo Chemical Physics Letters vol 380 no 1-2 pp 42ndash462003

[134] C Tripisciano K Kraemer A Taylor and E Borowiak-PalenldquoSingle-wall carbon nanotubes based anticancer drug deliverysystemrdquoChemical Physics Letters vol 478 no 4ndash6 pp 200ndash2052009

[135] C Tripisciano S Costa R J Kalenczuk andE Borowiak-PalenldquoCisplatin filledmultiwalled carbon nanotubesmdasha novelmolec-ular hybrid of anticancer drug containerrdquo European PhysicalJournal B vol 75 no 2 pp 141ndash146 2010

[136] J Li S Q Yap S L Yoong et al ldquoCarbon nanotube bottlesfor incorporation release and enhanced cytotoxic effect ofcisplatinrdquo Carbon vol 50 no 4 pp 1625ndash1634 2012

[137] L Sui T Yang P Gao et al ldquoIncorporation of cisplatin intoPEG-wrapped ultrapurified large-inner-diameterMWCNTs forenhanced loading efficiency and release profilerdquo InternationalJournal of Pharmaceutics vol 471 no 1-2 pp 157ndash165 2014

[138] E C Gryparis M Hatziapostolou E Papadimitriou and KAvgoustakis ldquoAnticancer activity of cisplatin-loaded PLGA-mPEGnanoparticles on LNCaP prostate cancer cellsrdquoEuropeanJournal of Pharmaceutics and Biopharmaceutics vol 67 no 1pp 1ndash8 2007

[139] N Nishiyama and K Kataoka ldquoPreparation and character-ization of size-controlled polymeric micelle containing cis-dichlorodiammineplatinum(II) in the corerdquo Journal of Con-trolled Release vol 74 no 1ndash3 pp 83ndash94 2001

[140] Y Mizumura Y Matsumura T Hamaguchi et al ldquoCisplatin-incorporated polymeric micelles eliminate nephrotoxicitywhile maintaining antitumor activityrdquo Japanese Journal of Can-cer Research vol 92 no 3 pp 328ndash336 2001

[141] A Schroeder R Honen K Turjeman A Gabizon J Kost andY Barenholz ldquoUltrasound triggered release of cisplatin fromliposomes in murine tumorsrdquo Journal of Controlled Release vol137 no 1 pp 63ndash68 2009


[142] M S Newman G T Colbern P K Working C Engbers andM A Amantea ldquoComparative pharmacokinetics tissue distri-bution and therapeutic effectiveness of cisplatin encapsulatedin long-circulating pegylated liposomes (SPI-077) in tumor-bearingmicerdquoCancer Chemotherapy and Pharmacology vol 43no 1 pp 1ndash7 1999

[143] K Ajima M Yudasaka T Murakami A Maigne K Shibaand S Iijima ldquoCarbon nanohorns as anticancer drug carriersrdquoMolecular Pharmaceutics vol 2 no 6 pp 475ndash480 2005

[144] K Ajima T Murakami Y Mizoguchi et al ldquoEnhancement ofin vivo anticancer effects of cisplatin by incorporation insidesingle-wall carbon nanohornsrdquo ACS Nano vol 2 no 10 pp2057ndash2064 2008

[145] S Hampel D Kunze D Haase et al ldquoCarbon nanotubesfilled with a chemotherapeutic agent a nanocarrier mediatesinhibition of tumor cell growthrdquoNanomedicine vol 3 no 2 pp175ndash182 2008

[146] N Graf D R Bielenberg N Kolishetti et al ldquo1205721199071205733integrin-

targeted PLGA-PEG nanoparticles for enhanced anti-tumorefficacy of a Pt(IV) prodrugrdquo ACS Nano vol 6 no 5 pp 4530ndash4539 2012

[147] S Mitra U Gaur P C Ghosh and A N Maitra ldquoTumour tar-geted delivery of encapsulated dextran-doxorubicin conjugateusing chitosan nanoparticles as carrierrdquo Journal of ControlledRelease vol 74 no 1ndash3 pp 317ndash323 2001

[148] F Lince S Bolognesi B Stella D L Marchisio and F DosioldquoPreparation of polymer nanoparticles loadedwith doxorubicinfor controlled drug deliveryrdquo Chemical Engineering Researchand Design vol 89 no 11 pp 2410ndash2419 2011

[149] H Park J Yang J Lee S Haam I-H Choi and K-H YooldquoMultifunctional nanoparticles for combined doxorubicin andphotothermal treatmentsrdquo ACS Nano vol 3 no 10 pp 2919ndash2926 2009

[150] Y Matsumura M Gotoh K Muro et al ldquoPhase I and phar-macokinetic study of MCC-465 a doxorubicin (DXR) encap-sulated in PEG immunoliposome in patients with metastaticstomach cancerrdquo Annals of Oncology vol 15 no 3 pp 517ndash5252004

[151] A A Gabizon ldquoPegylated liposomal doxorubicin metamor-phosis of an old drug into a new form of chemotherapyrdquo CancerInvestigation vol 19 no 4 pp 424ndash436 2001

[152] K Kataoka T Matsumoto M Yokoyama et al ldquoDoxorubicin-loaded poly(ethylene glycol)-poly(120573-benzyl-L-aspartate)copolymer micelles their pharmaceutical characteristics andbiological significancerdquo Journal of Controlled Release vol 64no 1ndash3 pp 143ndash153 2000

[153] F Perche N R Patel and V P Torchilin ldquoAccumulationand toxicity of antibody-targeted doxorubicin-loaded PEG-PEmicelles in ovarian cancer cell spheroid modelrdquo Journal ofControlled Release vol 164 no 1 pp 95ndash102 2012

[154] A B Ebrahim Attia C Yang J P K Tan et al ldquoThe effect ofkinetic stability on biodistribution and anti-tumor efficacy ofdrug-loaded biodegradable polymeric micellesrdquo Biomaterialsvol 34 no 12 pp 3132ndash3140 2013

[155] A Cambon A Rey-Rico D Mistry et al ldquoDoxorubicin-loadedmicelles of reverse poly(butylene oxide)-poly(ethylene oxide)-poly(butylene oxide) block copolymers as efficient lsquoactiversquochemotherapeutic agentsrdquo International Journal of Pharmaceu-tics vol 445 no 1-2 pp 47ndash57 2013

[156] J M Koziara P R Lockman D D Allen and R J MumperldquoPaclitaxel nanoparticles for the potential treatment of brain

tumorsrdquo Journal of Controlled Release vol 99 no 2 pp 259ndash269 2004

[157] G Ruan and S-S Feng ldquoPreparation and characterization ofpoly(lactic acid)-poly(ethylene glycol)-poly(lactic acid) (PLA-PEG-PLA) microspheres for controlled release of paclitaxelrdquoBiomaterials vol 24 no 27 pp 5037ndash5044 2003

[158] C Fonseca S Simoes and R Gaspar ldquoPaclitaxel-loaded PLGAnanoparticles preparation physicochemical characterizationand in vitro anti-tumoral activityrdquo Journal of Controlled Releasevol 83 no 2 pp 273ndash286 2002

[159] J M Chan J W Rhee C L Drum et al ldquoIn vivo prevention ofarterial restenosis with paclitaxel-encapsulated targeted lipid-polymeric nanoparticlesrdquo Proceedings of the National Academyof Sciences of the United States of America vol 108 no 48 pp19347ndash19352 2011

[160] T Hamaguchi K Kato H Yasui et al ldquoA phase I and pharma-cokinetic study of NK105 a paclitaxel-incorporating micellarnanoparticle formulationrdquo British Journal of Cancer vol 97 no2 pp 170ndash176 2007

[161] T-Y Kim D-W Kim J-Y Chung et al ldquoPhase I and pharma-cokinetic study of Genexol-PM a Cremophor-free polymericmicelle-formulated paclitaxel in patients with advanced malig-nanciesrdquo Clinical Cancer Research vol 10 no 11 pp 3708ndash37162004

[162] F Wang Y Chen D Zhang et al ldquoFolate-mediated targetedand intracellular delivery of paclitaxel using a novel deoxycholicacid-O-carboxymethylated chitosan-folic acid micellesrdquo Inter-national Journal of Nanomedicine vol 7 pp 325ndash337 2012

[163] Z Liu J T Robinson S M Tabakman K Yang and HDai ldquoCarbon materials for drug delivery and cancer therapyrdquoMaterials Today vol 14 no 7-8 pp 316ndash323 2011

[164] K R West and S Otto ldquoReversible covalent chemistry in drugdeliveryrdquo Current Drug Discovery Technologies vol 2 no 3 pp123ndash160 2005

[165] C Li R A Newman Q-P Wu et al ldquoBiodistribution ofpaclitaxel and poly(L-glutamic acid)-paclitaxel conjugate inmice with ovarian OCa-1 tumorrdquo Cancer Chemotherapy andPharmacology vol 46 no 5 pp 416ndash422 2000

[166] L Milas K A Mason N Hunter C Li and S Wal-lace ldquoPoly(L-glutamic acid)-paclitaxel conjugate is a potentenhancer of tumor radiocurabilityrdquo International Journal ofRadiation Oncology Biology Physics vol 55 no 3 pp 707ndash7122003

[167] T Etrych T Mrkvan P Chytil C Konak B Rıhova andK Ulbrich ldquoN-(2-hydroxypropyl)methacrylamide-based poly-mer conjugates with pH-controlled activation of doxorubicinI New synthesis physicochemical characterization and prelim-inary biological evaluationrdquo Journal of Applied Polymer Sciencevol 109 no 5 pp 3050ndash3061 2008

[168] M Hruby C Konak and K Ulbrich ldquoPolymeric micellarpH-sensitive drug delivery system for doxorubicinrdquo Journal ofControlled Release vol 103 no 1 pp 137ndash148 2005

[169] D Vetvicka M Hruby O Hovorka et al ldquoBiological evaluationof polymeric micelles with covalently bound doxorubicinrdquoBioconjugate Chemistry vol 20 no 11 pp 2090ndash2097 2009

[170] C Liu F Liu L Feng M Li J Zhang and N Zhang ldquoThetargeted co-delivery of DNA and doxorubicin to tumor cells viamultifunctional PEI-PEG based nanoparticlesrdquo Biomaterialsvol 34 no 10 pp 2547ndash2564 2013

[171] H S Yoo E A Lee and T G Park ldquoDoxorubicin-conjugatedbiodegradable polymeric micelles having acid-cleavable link-agesrdquo Journal of Controlled Release vol 82 no 1 pp 17ndash27 2002


[172] S Aryal C-M J Hu and L Zhang ldquoPolymer-cisplatin con-jugate nanoparticles for acid-responsive drug deliveryrdquo ACSNano vol 4 no 1 pp 251ndash258 2010

[173] E Tsai ldquoDendrimer encapsulated nanoparticles vehicles fordrug delivery with an emphasis on PAMAM dendrimersrdquoCosmos vol 8 pp 1ndash16 2011

[174] P C A Rodrigues K Scheuermann C Stockmar et alldquoSynthesis and in vitro efficacy of acid-sensitive poly(ethyleneglycol) paclitaxel conjugatesrdquo Bioorganic and Medicinal Chem-istry Letters vol 13 no 3 pp 355ndash360 2003

[175] R P Feazell N Nakayama-Ratchford H Dai and S J LippardldquoSoluble single-walled carbon nanotubes as longboat deliverysystems for platinum(IV) anticancer drug designrdquo Journal of theAmerican Chemical Society vol 129 no 27 pp 8438ndash8439 2007

[176] Y-Z Zhao C-Z Sun C-T Lu et al ldquoCharacterization andanti-tumor activity of chemical conjugation of doxorubicin inpolymeric micelles (DOX-P) in vitrordquo Cancer Letters vol 311no 2 pp 187ndash194 2011

[177] P-S Lai P-J Lou C-L Peng et al ldquoDoxorubicin delivery bypolyamidoamine dendrimer conjugation and photochemicalinternalization for cancer therapyrdquo Journal of Controlled Releasevol 122 no 1 pp 39ndash46 2007

[178] J Chen S Chen X Zhao L V Kuznetsova S S Wong andI Ojima ldquoFunctionalized single-walled carbon nanotubes asrationally designed vehicles for tumor-targeted drug deliveryrdquoJournal of the American Chemical Society vol 130 no 49 pp16778ndash16785 2008

[179] L Niu L Meng and Q Lu ldquoFolate-conjugated PEG on singlewalled carbon nanotubes for targeting delivery of doxorubicinto cancer cellsrdquo Macromolecular Bioscience vol 13 no 6 pp735ndash744 2013

[180] K Kataoka T Matsumoto M Yokoyama et al ldquoDoxorubicin-loaded poly(ethylene glycol)-poly(beta-benzyl-L-aspartate)copolymer micelles their pharmaceutical characteristics andbiological significancerdquo Journal of Controlled Release vol 64no 1ndash3 pp 143ndash153 2000

[181] K Greish T Sawa J Fang T Akaike and H Maeda ldquoSMA-doxorubicin a new polymeric micellar drug for effective tar-geting to solid tumoursrdquo Journal of Controlled Release vol 97no 2 pp 219ndash230 2004

[182] H Ali-Boucetta K T Al-Jamal D McCarthy M Prato ABianco and K Kostarelos ldquoMultiwalled carbon nanotube-doxorubicin supramolecular complexes for cancer therapeu-ticsrdquoChemical Communications vol 8 no 4 pp 459ndash461 2008

[183] E Heister V Neves C Tılmaciu et al ldquoTriple functionalisationof single-walled carbon nanotubes with doxorubicin a mono-clonal antibody and a fluorescent marker for targeted cancertherapyrdquo Carbon vol 47 no 9 pp 2152ndash2160 2009

[184] X Zhang L Meng Q Lu Z Fei and P J Dyson ldquoTargeteddelivery and controlled release of doxorubicin to cancer cellsusingmodified single wall carbon nanotubesrdquo Biomaterials vol30 no 30 pp 6041ndash6047 2009

[185] Z Liu X Sun N Nakayama-Ratchford and H DaildquoSupramolecular chemistry on water-soluble carbon nanotubesfor drug loading and deliveryrdquo ACS Nano vol 1 no 1 pp50ndash56 2007

[186] A Di Crescenzo D Velluto J A Hubbell and A FontanaldquoBiocompatible dispersions of carbon nanotubes a potentialtool for intracellular transport of anticancer drugsrdquo Nanoscalevol 3 no 3 pp 925ndash928 2011

[187] Z Tian Y ShiM YinH Shen andN Jia ldquoFunctionalizedmul-tiwalled carbon nanotubes-anticancer drug carriers synthesis

targeting ability and antitumor activityrdquo Nano Biomedicine andEngineering vol 3 no 3 pp 157ndash162 2011

[188] A Duma M Prodana and I Demetrescu ldquoCisplatin func-tionalization of multiwall carbon nanotubesrdquo UPB ScientificBulletin Series B Chemistry and Materials Science vol 76 no1 pp 49ndash58 2014

[189] A Barroug L T Kuhn L C Gerstenfeld and M J GlimcherldquoInteractions of cisplatin with calcium phosphate nanoparti-cles in vitro controlled adsorption and releaserdquo Journal ofOrthopaedic Research vol 22 no 4 pp 703ndash708 2004

[190] M Arlt D Haase S Hampel et al ldquoDelivery of carboplatinby carbon-based nanocontainers mediates increased cancer celldeathrdquo Nanotechnology vol 21 no 33 Article ID 335101 2010

[191] Y Wang S-T Yang Y Wang Y Liu and H Wang ldquoAdsorptionand desorption of doxorubicin on oxidized carbon nanotubesrdquoColloids and Surfaces B Biointerfaces vol 97 pp 62ndash69 2012

[192] Y-I Jeong S-T Kim S-G Jin et al ldquoCisplatin-lncorporatedhyaluronic acid nanoparticles based on ion-complex forma-tionrdquo Journal of Pharmaceutical Sciences vol 97 no 3 pp 1268ndash1276 2008

[193] T K Bronich A Nehls A Eisenberg V A Kabanov and A VKabanov ldquoNovel drug delivery systems based on the complexesof block ionomers and surfactants of opposite chargerdquo Colloidsand Surfaces B Biointerfaces vol 16 no 1ndash4 pp 243ndash251 1999

[194] G Pastorin W Wu S Wieckowski et al ldquoDouble function-alisation of carbon nanotubes for multimodal drug deliveryrdquoChemical Communications no 11 pp 1182ndash1184 2006

[195] R Li R Wu L Zhao M Wu L Yang and H Zou ldquoP-glycoprotein antibody functionalized carbon nanotube over-comes the multidrug resistance of human leukemia cellsrdquo ACSNano vol 4 no 3 pp 1399ndash1408 2010

[196] L Zhang A F Radovic-Moreno F Alexis et al ldquoCo-delivery ofhydrophobic and hydrophilic drugsfrom nanoparticle-aptamerbioconjugatesrdquo ChemMedChem vol 2 no 9 pp 1268ndash12712007

[197] K Werengowska-Ciecwierz M Wisniewski A P Terzyk et alldquoNanotube-mediated efficiency of cisplatin anticancer therapyrdquoCarbon vol 70 pp 46ndash58 2014

[198] N Lodhi N K Mehra and N K Jain ldquoDevelopment andcharacterization of dexamethasonemesylate anchored onmultiwalled carbon nanotubesrdquo Journal of Drug Targeting vol 21 no1 pp 67ndash76 2013

[199] J Shi Z Xiao N Kamaly andO C Farokhzad ldquoSelf-assembledtargeted nanoparticles evolution of technologies and bench tobedside translationrdquo Accounts of Chemical Research vol 44 no10 pp 1123ndash1134 2011

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

High Energy PhysicsAdvances in

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


FluidsJournal of

Atomic and Molecular Physics

Journal of



Advances in Condensed Matter Physics

OpticsInternational Journal of



AstronomyAdvances in

International Journal of


Superconductivity


Statistical MechanicsInternational Journal of


GravityJournal of


AstrophysicsJournal of


Physics Research International


Solid State PhysicsJournal of

Computational Methods in Physics

Journal of



Soft MatterJournal of

Hindawi Publishing Corporationhttpwwwhindawicom

AerodynamicsJournal of

Volume 2014


PhotonicsJournal of


Journal of

Biophysics


ThermodynamicsJournal of


Classical therapy Targeted therapy

Cancer cell Endothelial cell

Nanocarrier Antibody nanocarrier

Highconcentration

Lowconcentration

ligand to receptorHigh affinity ofEPR effect

Figure 1 The mechanism of drug delivery in classical and targeted therapy (the figure is based on [12 199])

liposomes magnetic nanoparticles and carbon nanostruc-tures are extensively studied [4ndash10] All theseDDSs have beenreported to have huge biomedical potential

Nanomedicine is a rapidly developing research areain biological and medical science and the development ofnanomedicine research requires collaboration among dif-ferent scientists physicians engineers molecular biologistsmaterial scientists chemists and so forth [8] Applicationsof nanotechnology in medicine are especially promisingand areas such as disease diagnosis drug delivery targetedat specific sites in the body (see the next paragraph) andmolecular imaging are intensively investigated and someproducts are undergoing clinical trials Nanotechnology playsan important role in therapies of the future as ldquonanomedicinerdquoby lowering doses required for efficacy as well as increasingthe therapeutic indices and safety profiles of new therapeutics[9] The therapeutics are delivery systems in the nanometersize range containing encapsulated dispersed adsorbed orconjugated drugs and imaging agents Selected nanoscalesystems including liposomes micelles nanoemulsionsnanoparticulate systems (drug nanoparticles polymer-lipid- carbon- and ceramic-based albumin and nanogels)and dendrimers are used for drug delivery and imaging [9]

Within the next 5 to 10 years nanomedicine will addressmany important medical problems by using nanoscale-structured materials and simple nanodevices that can bemanufactured today Many approaches to nanomedicinebeing carried out today are already close enough to fruitionthat it is fair to say that their successful development isalmost inevitable and their subsequent incorporation intovaluable medical diagnostics or clinical therapeutics ishighly likely and may occur very soon Among them Fre-itas Jr [4] mentioned immunoisolation gated nanosievesultrafast DNA sequencing fullerene-based pharmaceuticalsnanoshells single-virus detectors tectodendrimers radio-controlled biomolecules and biologic robots [4] Flynn andWei [10] mentioned also the importance of commercializa-tion of nanomedicine

In the next chapter we present short review on one ofthe practical applications of nanotechnology inmedicineWefocus on the application of nanoparticles as DDS

2 Targeted Anticancer Therapy

A deeper understanding of the molecular events leading totumor formation invasion angiogenesis and metastasis hasprovided a newmechanistic basis for drug discovery targetedanticancer therapy By specifically blocking the molecularpathways implicated in the pathogenesis of cancer targetedanticancer agents are expected to alter the natural courseof the disease and at the same time to offer an enhancedtherapeutic index over traditional cytotoxic agents

Targeted anticancer therapy must fulfill few requestsAnticancer drugs should be delivered to the cancer cellswith minimal activity loss The drug should kill selectivelythe cancer cells The release of drug active form must beof course controlled [11] Simultaneously only the minimaldoses of chemotherapeutic agent should be administeredduring the targeted therapy (doses should be smaller thanthose in the traditional chemotherapy) The therapy shouldalso minimize different possible side effects [11 12]

Currently drugs are usually delivered inside or outsideof nanocarriers via the passive or selective mechanism inconventional or targeted therapy respectively [12] (Figure 1)The traditional mechanism is connected with a drug aggre-gation process (in form of carrier drug) inside cancer tissuesvia enhanced permeability and retention effect (EPR) [13ndash15] This method is based on the abnormal structure ofblood vessels near tumor Thus the drug reaches easily thetissues near cancer cells [9 14] The most popular drugsused during conventional treatment are doxorubicin [16]paclitaxel [17] methotrexate [18] hexamethylmelamine [19]and gemcitabine [20] and drugs based on platinum [21] likecisplatin (DDP) or carboplatin (Figure 2) The linking ofdrugs to nanocarriers is described in this paper together withprogressive activity after linking with antibody


OHOH

O

ONH

O

OO

AcO O OH

OHAcO

(a)

OO

O OH O

O

OH

OH

OH

OCH3

NH2

CH2OH

(b)

N N

NN N

H2N

NH2NH

OH

OH

O

O

O

(c)

N

N

O O

F

FOH

HO

NH2

(d)

PtOCO

OCO

H3N

H3N

(e)

PtCl

ClH3N

H3N

(f)

N

N

N

N

N

N

(g)































(a) (b)

(c) (d) (e) (f)


















O OO

ONH

N

NEDC

OH

(CH2)3 NH2HN

Scheme 1














HO

OO O O

O

OO

O

O

O

OO

O

O

O

O

OO

O O

O

O HN

O

O

O

HO

S

O NH NH

NH

OH

P

H

O O

O

O

N

S

OH

O

O

O

OH

N

OH

O O

O

P

P

O

O

(OCH2CH2)45

(OCH2CH2)45

(OCH2CH2)45

NH

NH

HO

+ EDCNHS

Protein A

Protein A

NH2

NH

NH2 +

Protein AOminus

Ominus

Ominus

Scheme 2





DSPNH

O

SS

O

O

NO O

NHS

NH

OS

S

O

NH

NH2

NH2

Scheme 3

N

O

O

N

O

OS

SH +

R1

R1 R2R2C1C1

Scheme 4


















n

n

NN

N

O

O

O

O

O

OO

O

O

OSH

S

H

NH

N

HOOC

HOOC

COOH

COOH

H

NH

NH2+Clminus

NH2+Clminus

NH2

NH2

NH2

NH2

Scheme 5

NO O

NH

O

SH NH

O

S

N

OO

OH

O

EDC DTT SPDP D

TTNH2

Scheme 6

S SR1R1 R2R2SH + HS

Scheme 7

NH NH N

O

O

OH

NH2

Scheme 8




O

O

N

O

ON

O

O

Scheme 9




H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

















XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




OHOH

O

ONH

O

OO

AcO O OH

OHAcO

(a)

OO

O OH O

O

OH

OH

OH

OCH3

NH2

CH2OH

(b)

N N

NN N

H2N

NH2NH

OH

OH

O

O

O

(c)

N

N

O O

F

FOH

HO

NH2

(d)

PtOCO

OCO

H3N

H3N

(e)

PtCl

ClH3N

H3N

(f)

N

N

N

N

N

N

(g)































(a) (b)

(c) (d) (e) (f)


















O OO

ONH

N

NEDC

OH

(CH2)3 NH2HN

Scheme 1














HO

OO O O

O

OO

O

O

O

OO

O

O

O

O

OO

O O

O

O HN

O

O

O

HO

S

O NH NH

NH

OH

P

H

O O

O

O

N

S

OH

O

O

O

OH

N

OH

O O

O

P

P

O

O

(OCH2CH2)45

(OCH2CH2)45

(OCH2CH2)45

NH

NH

HO

+ EDCNHS

Protein A

Protein A

NH2

NH

NH2 +

Protein AOminus

Ominus

Ominus

Scheme 2





DSPNH

O

SS

O

O

NO O

NHS

NH

OS

S

O

NH

NH2

NH2

Scheme 3

N

O

O

N

O

OS

SH +

R1

R1 R2R2C1C1

Scheme 4


















n

n

NN

N

O

O

O

O

O

OO

O

O

OSH

S

H

NH

N

HOOC

HOOC

COOH

COOH

H

NH

NH2+Clminus

NH2+Clminus

NH2

NH2

NH2

NH2

Scheme 5

NO O

NH

O

SH NH

O

S

N

OO

OH

O

EDC DTT SPDP D

TTNH2

Scheme 6

S SR1R1 R2R2SH + HS

Scheme 7

NH NH N

O

O

OH

NH2

Scheme 8




O

O

N

O

ON

O

O

Scheme 9




H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

















XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics



























(a) (b)

(c) (d) (e) (f)


















O OO

ONH

N

NEDC

OH

(CH2)3 NH2HN

Scheme 1














HO

OO O O

O

OO

O

O

O

OO

O

O

O

O

OO

O O

O

O HN

O

O

O

HO

S

O NH NH

NH

OH

P

H

O O

O

O

N

S

OH

O

O

O

OH

N

OH

O O

O

P

P

O

O

(OCH2CH2)45

(OCH2CH2)45

(OCH2CH2)45

NH

NH

HO

+ EDCNHS

Protein A

Protein A

NH2

NH

NH2 +

Protein AOminus

Ominus

Ominus

Scheme 2





DSPNH

O

SS

O

O

NO O

NHS

NH

OS

S

O

NH

NH2

NH2

Scheme 3

N

O

O

N

O

OS

SH +

R1

R1 R2R2C1C1

Scheme 4


















n

n

NN

N

O

O

O

O

O

OO

O

O

OSH

S

H

NH

N

HOOC

HOOC

COOH

COOH

H

NH

NH2+Clminus

NH2+Clminus

NH2

NH2

NH2

NH2

Scheme 5

NO O

NH

O

SH NH

O

S

N

OO

OH

O

EDC DTT SPDP D

TTNH2

Scheme 6

S SR1R1 R2R2SH + HS

Scheme 7

NH NH N

O

O

OH

NH2

Scheme 8




O

O

N

O

ON

O

O

Scheme 9




H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

















XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics
















O OO

ONH

N

NEDC

OH

(CH2)3 NH2HN

Scheme 1














HO

OO O O

O

OO

O

O

O

OO

O

O

O

O

OO

O O

O

O HN

O

O

O

HO

S

O NH NH

NH

OH

P

H

O O

O

O

N

S

OH

O

O

O

OH

N

OH

O O

O

P

P

O

O

(OCH2CH2)45

(OCH2CH2)45

(OCH2CH2)45

NH

NH

HO

+ EDCNHS

Protein A

Protein A

NH2

NH

NH2 +

Protein AOminus

Ominus

Ominus

Scheme 2





DSPNH

O

SS

O

O

NO O

NHS

NH

OS

S

O

NH

NH2

NH2

Scheme 3

N

O

O

N

O

OS

SH +

R1

R1 R2R2C1C1

Scheme 4


















n

n

NN

N

O

O

O

O

O

OO

O

O

OSH

S

H

NH

N

HOOC

HOOC

COOH

COOH

H

NH

NH2+Clminus

NH2+Clminus

NH2

NH2

NH2

NH2

Scheme 5

NO O

NH

O

SH NH

O

S

N

OO

OH

O

EDC DTT SPDP D

TTNH2

Scheme 6

S SR1R1 R2R2SH + HS

Scheme 7

NH NH N

O

O

OH

NH2

Scheme 8




O

O

N

O

ON

O

O

Scheme 9




H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

















XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




HO

OO O O

O

OO

O

O

O

OO

O

O

O

O

OO

O O

O

O HN

O

O

O

HO

S

O NH NH

NH

OH

P

H

O O

O

O

N

S

OH

O

O

O

OH

N

OH

O O

O

P

P

O

O

(OCH2CH2)45

(OCH2CH2)45

(OCH2CH2)45

NH

NH

HO

+ EDCNHS

Protein A

Protein A

NH2

NH

NH2 +

Protein AOminus

Ominus

Ominus

Scheme 2





DSPNH

O

SS

O

O

NO O

NHS

NH

OS

S

O

NH

NH2

NH2

Scheme 3

N

O

O

N

O

OS

SH +

R1

R1 R2R2C1C1

Scheme 4


















n

n

NN

N

O

O

O

O

O

OO

O

O

OSH

S

H

NH

N

HOOC

HOOC

COOH

COOH

H

NH

NH2+Clminus

NH2+Clminus

NH2

NH2

NH2

NH2

Scheme 5

NO O

NH

O

SH NH

O

S

N

OO

OH

O

EDC DTT SPDP D

TTNH2

Scheme 6

S SR1R1 R2R2SH + HS

Scheme 7

NH NH N

O

O

OH

NH2

Scheme 8




O

O

N

O

ON

O

O

Scheme 9




H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

















XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




DSPNH

O

SS

O

O

NO O

NHS

NH

OS

S

O

NH

NH2

NH2

Scheme 3

N

O

O

N

O

OS

SH +

R1

R1 R2R2C1C1

Scheme 4


















n

n

NN

N

O

O

O

O

O

OO

O

O

OSH

S

H

NH

N

HOOC

HOOC

COOH

COOH

H

NH

NH2+Clminus

NH2+Clminus

NH2

NH2

NH2

NH2

Scheme 5

NO O

NH

O

SH NH

O

S

N

OO

OH

O

EDC DTT SPDP D

TTNH2

Scheme 6

S SR1R1 R2R2SH + HS

Scheme 7

NH NH N

O

O

OH

NH2

Scheme 8




O

O

N

O

ON

O

O

Scheme 9




H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

















XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




n

n

NN

N

O

O

O

O

O

OO

O

O

OSH

S

H

NH

N

HOOC

HOOC

COOH

COOH

H

NH

NH2+Clminus

NH2+Clminus

NH2

NH2

NH2

NH2

Scheme 5

NO O

NH

O

SH NH

O

S

N

OO

OH

O

EDC DTT SPDP D

TTNH2

Scheme 6

S SR1R1 R2R2SH + HS

Scheme 7

NH NH N

O

O

OH

NH2

Scheme 8




O

O

N

O

ON

O

O

Scheme 9




H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

















XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




H O O O OO

H

O

COOH

O

OO NH

OO NH

O

O

1 24 169

71

Self-assembly

N

O

O

O

O

O

O

O

O

O O

N OO

OO

N

O

O

Scheme 10

RCH

NN

N

R

Cu (I)N3 + R1

R1

Cycloaddition

Scheme 11

















XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics









XHX

NuNu


H3O+

X = O NR +SR +NR2

∙∙

Scheme 12


R3X NH2R1 R1

R2R2

O N XR3

+ H2O X = O NR

Scheme 13

R3

R3

NH2

NH

R1 R1

R2

O O

+ HR2

Scheme 14




Ligand nanocarrier

Drug conjugation

Drug entrapment





R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




R1

R1

R2

R2



Scheme 15

EWG1

EWG1

EWG2

EWG2EWG3

EWG3

+ H2C

Scheme 16

CuLL

LCuCu H

H

L

LCuCu

CuL

LCu

H

H

B

B

L

LCuCu

NN

N

NN

N

N

Cu

N

L

LCu

Metallacycle

R1

R1

R1

R1

R1

Bminus

Bminus

R2

R2

R2

R2

R1

N3

N+

Scheme 17











SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics






SWCNT


Nanocondensation

C60

CNTs

Solvent

ldquoGuestrdquo

Wea

k Weak

Strong

Filtration paper

TEM grid

SWCNTs

C60-toluene

CNTs

Solvent

ldquoGuestrdquoStr

ong Strong

Strong

SWCNTToluene








Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




Drug

PLGA

Au layer





PEG outer shell


PTXBlock copolymer











O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




O OH

OH

O

OH

OMe O OH

O

OH

Fmoc-OSu DMF

O OH

OH

O

OH

OMe O OH

O

NHFmocOH

O

O

OH

OH

O

O

OH

OMe

OO

H

O

O

O

NHFmocOH

OH

OH

n

Piperidine DMF

O

O

O

OH

OMe O

OO

H

O

O

O

NH2OH

n

CH2

CH2

CH2

CH2

NH2

H3C

PLGA5005 PyBroP

DMAP TEA CH2Cl2

Scheme 18






C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




C

O

O OOO

OH

O

O

O

O

OOO

O OH

O

O

O

O

C

O

O

DOXDMF TEA

O

OH

OH

OH

O

OH

OMe O

O

NHOH

OOO

O OH

O

O

O

O

O

n

n

n

Cl + HOO2N

O2NPyridine CH2Cl2

Scheme 19






2with




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics




O OHO

O

OOHO

O

S

OO

OHOH

OH

OHOMe OO

OH

OO

O

S

NHON

OOOHO

O

S

NHO

OH

O

S

NHO

DOX

nnm minus xxmnmnm

NH2NH2

NH2

OCH3

N2H4HSCH2COOCH3

Scheme 20



3)2Cl2(OEt)(O






















6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics













6 Summary



Abbreviations














Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics















Acknowledgment


References



























































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics













































































































































































































FluidsJournal of


Journal of










Superconductivity




GravityJournal of








Journal of






Volume 2014


PhotonicsJournal of


Journal of

Biophysics



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