Materials Science & Engineering C 106 (2020) 110302

Contents lists available at ScienceDirect

Materials Science & Engineering C

journal homepage: www.elsevier.com/locate/msec

Grand challenges in nanomedicine T

a,∗,1 b,1 c
Lin-Ping Wu , Danyang Wang , Zibiao Li
a
Drug Discovery Pipeline, Hefei Institute of Stem Cell and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences,
Guangzhou, 510530, China
b
Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200, Copenhagen,
Denmark
c
Institute of Materials Research and Engineering, A*STAR (Agency for Science, Technology and Research), 2 Fusionopolis Way, Innovis, #08-03, Singapore

A R T I C LE I N FO A B S T R A C T

Keywords: Nanotherapeutics and nanopharmaceuticals could achieve and facilitate earlier and more precise individual
Nanomedicine diagnosis, improve targeted therapies, reduce side effects, and enhance therapeutic monitoring. These ad-
Nanoparticles vantages will improve quality of life, support a healthier and more independent aging population, and be in-
Drug delivery strumental in maximizing the cost-effectiveness of health care. However, the field of nanomedicine is at its early
Translation medicine
stage, most of the research still stays in the laboratory phase, and few success stories are translated into clinical
Diagnostics
trials and medical practice. This review will demonstrate the numerous challenges that are encountered during
the development of commercial nanoparticle-based therapeutics and the possible solutions.

1. Introduction and reproducible scale-up and manufacturing process to achieve a

In recent years, nanotechnology has been increasingly applied to the
area of medicine, which is defined as nanomedicine, as a new in-
dependent field of life sciences [1,2]. Nanomedicine has the potential
advantages to overcome biological barriers, effectively deliver hydro-
phobic drugs and biologics, and preferentially target sites of disease
[3,4]. However, only a relatively small number of nanoparticle-based
medicines have been approved for clinical use because of numerous
challenges and hurdles at different stages of development [5,6]. The
first FDA-approved nano-drug is Doxil® in 1995 (Fig. 1), which can
prolong drug circulation time and is avoidance of the RES due to the use
of PEGylated nano-liposomes. What's more important, doxorubicin is
highly and stably remote loaded in Doxil® using a transmembrane
ammonium sulfate gradient method, which also allows for drug release
at the tumor [7]. The promising nanomedicine approved by FDA or in
the clinical trials has been summarized by lots of reviews [8–11]. From
1995 to 2017, 50 nano-pharmaceuticals have received FDA approval
and are currently available for clinical use [10]. Also as of August 2019,
64 clinical trials including the term “nano” were listed as “recruiting”
or “active” on ClinicalTrials.gov. The attractiveness of nanomedicine
applications lies in the unique characteristics of three-dimensional
constructs with multiple components in nanoscale. At the same time,
due to its complexity, nanomedicine product requires careful design
and engineering, strict characterization of physicochemical properties,
consistent product with stable physicochemical characteristics, biolo-
gical behaviors, and pharmacological profiles. The safety and reg-
ulatory issues of nanomedicine need additional considerations com-
pared with conventional medicines. This review seeks to summary
challenges and limitations during the development and commerciali-
zation of nanomedicine products and discusses potential solutions to
accelerate the growth of this important field.
2. Challenges and current limitations
Successfully translating nanomedicine from pre-clinical proof of
concept to demonstration of therapeutic value in the clinic is still
challenging, several obstacles have been identified as top scientific
hurdles in bringing nanoengineered products to patients (Fig. 2).
2.1. Challenges of biological barriers and specific targeting
Before the drugs successfully reach the disease sites, the nano-for-
mulation has to help them to cross multiple biological barriers (Fig. 3)
[6,12]. For example, oral nanoparticles need to have high stability in
the gastrointestinal tract, the ability to penetrate intestinal epithelium
and the ability to keep the high systemic bioavailability of drugs after
crossing several barriers [12,13].
Compared with most of the small molecules delivered orally, large

∗
Corresponding author.
E-mail address: wu_linping@gibh.ac.cn (L.-P. Wu).
1
Contributed equally.

https://doi.org/10.1016/j.msec.2019.110302
Received 25 August 2018; Received in revised form 10 October 2019; Accepted 10 October 2019
Available online 12 October 2019
0928-4931/ © 2019 Elsevier B.V. All rights reserved.
L.-P. Wu, et al.
Fig. 2. The challenges of safe translation nanomedicine products for diagnostics and treatment in clinic.
molecular drugs, such as peptides, proteins, and polynucleotides,
usually adopt intravenous administration (IV) which remains the most
direct and efficient route to deliver therapeutic reagents [14]. After IV
injection, drugs in circulation still need to overcome several biological
barriers to reach their targets. For the treatment of central nervous
system related disease, the blood-brain barrier (BBB) restricts the dif-
fusion of 99% large or hydrophilic molecules into the cerebrospinal
fluid and is a major obstacle for brain targeting delivery [15–17]. There
are various possible transport pathways by which solute molecules and
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Materials Science & Engineering C 106 (2020) 110302
Fig. 1. The first FDA-approved nano-drug Doxil®.
(A) Doxil® vial as sold by Sequus Pharmaceuticals
(starting 1996), (B) cryo-TEMs of commercial
Doxil, which is doxorubicin sulfate remote loaded
pegylated nano-liposomes, (C) a sterically stabi-
lized (PEGylated) liposome, shows lack of inser-
tion of plasma opsonin into its membrane (Reprint
from Ref. [7]).
Fig. 3. Typical extremely effective biological
barriers largely prevent nanoparticles from
reaching their intended destinations. The barriers
include the reticuloendothelial system, en-
dothelial/epithelial membranes, complex net-
works of blood vessels, abnormal flow of blood,
and interstitial pressure gradients. Inset: A mag-
nified region of a tumor cell offers additional
biological challenges: cellular/nuclear membranes
and ionic/molecular pumps that can expel drugs
from the cancerous cells creating drug resistance
(Reprint from Ref. [6]).
nanoparticles move across the BBB, as shown in Fig. 4 [15]. Another
major challenge of nanomedicine is to deliver therapeutic reagents into
solid tumors [18,19]. Although tumor vasculature is high hetero-
geneous in distribution and more permeable in some places, large areas
of tumors with high cancer cell density and dense tumor stroma is still
poor perfused which further hamper the distribution of drugs in tumors
[20]. Increased interstitial fluid pressure (IFP) caused by impaired
lymphatic drainage in tumors [21,22] adds another barrier to delivery,
which is one of the main factors contributing to reduce extravasation
L.-P. Wu, et al.
Fig. 4. Transport pathways across BBB (Reprint from Ref. [15]).
and transvascular transport of drugs despite the leaky tumor micro-
vasculature, and restrain the transport of molecules into interstitial
space of tumor [23]. Facing these challenges, multiple nanoparticle-
based strategies from biomaterials, including liposome, polymer mi-
celle, and peptidic or protein nanoparticles, are investigated to deliver
drugs [24–26]. Because of the high permeability of tumor vasculature
and lack of proper lymphatic drainage, the enhanced permeability and
retention effect (EPR) enhance the passive accumulation of nano-
particles and macromolecules in the tumor microenvironment and im-
prove tumor drug delivery [22]. First generation nanomedicine on the
market is based on passive targeting, such as pegylated liposomal
doxorubicin (Doxil®/Caelyx®) [7,27] and nab-paclitaxel (Abraxane®)
[28]. The key issue of passively targeted first generation nanomedicine
is how to control the pharmacokinetics and biodistribution of nano-
particles by modulating its physicochemical properties [29].
Even though there are some advantages of the passive process, the
utilization of an active biological transport process can increase effi-
cacy, decrease side effects, and reduce systemic drug exposures, which
further improve drug delivery [30]. The organized structures of active
targeting nanoparticles allow for the incorporation with various tar-
geting moieties including small molecular ligands for receptors, pep-
tides, proteins, antibodies, aptamers, and oligonucleotides. It can en-
hance drug delivery to the target sites, reduce off-target organ
toxicities, and facilitate cellular uptake of therapeutic agents [31,32].
Usually the ligands or monoclonal antibodies targeting to the surface
receptors overexpressed by cancer cells, such as transferrin receptor
(TfR), folate receptor (FR) and epidermal growth factor receptor
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Materials Science & Engineering C 106 (2020) 110302
(EGFR), decorate the surface of nanoparticles to increase cellular in-
ternalization of the reagents through endocytosis and improve the ef-
ficacy of systemic anticancer therapy [33,34]. In addition, nano-
particles incorporating with protein transduction domain (PTD) and
cell penetrating peptides (CPPs), a class of small cationic peptides
(< 20 amino acid), such as oligoarginine and TAT, enable the uniform
transport of large, biologically functional molecules that otherwise
cannot effectively enter cancer cells [35,36]. Moreover, another pos-
sible target of nanomedicine is tumor endothelial cells. For example,
the cyclic and linear derivatives of oligopeptides RGD (Arg-Gly-Asp)
which bind to the integrins α2β3, αvb3, and α5β1, provide a tumor-
penetrating function to polymers, liposomes and other nanoparticles.
With RGD modification, nanoparticles deliver cytotoxic reagents to
tumor tissue and achieve significant antitumor effects [37–39]. The
active targeting depends on the affinity and efficacy of a target and its
specific ligand. Besides, it is very important to optimize the density of
targeting ligands per nanocarrier to achieve not only high targeting
efficiency but also to ensure an optimal internalization [40].
2.2. Analysis and characterization of nano-formulations
Compared with traditional pharmaceuticals, nanomedicine is a
three-dimensional complex composed of several components and each
component serves to a specific function [41]. Therefore, it requires
more sophisticated and appropriate analytical tests to fully detect,
characterize and quantify each component and also evaluate the re-
lationships and interactions between these components including both
L.-P. Wu, et al.
physicochemical properties and biological behaviors [28]. But it is one
of the challenges in the development of nanomedicine not only the
technical aspect but also the regulatory perspective. In general, the
most important physicochemical features of nanomedicine are compo-
sition, structure, size, surface properties, porosity, charge, and stability
[42,43]. It is difficult to characterize nanomedicine products because of
their variability properties. Taking polydispersity (PD) as an example, it
is a key parameter that measures the heterogeneity of particles with
regard to size, shape, or mass. If the nanomedicine product with the
same average size but different PD, it may cause dramatic changes in
secondary properties such as targeting properties, drug release rate,
biocompatibility, toxicity, and in vivo behaviors [44–47]. Another
challenge is the characterization of stability and storage of nanomedi-
cine. Because of the excellent features of biodegradable materials, such
as lipid and polymer, they have been widely used in the development of
nanomedicine products. But this biodegradable property of nanoma-
terials would also alter the properties of nanomedicine products during
the storage either in the solutions or in a lyophilized powder form
[48,49]. So it is substantial to improve quality assessment of the bio-
degradable materials by well-defined and reproducible standards. An-
other issue is detecting and visualizing the in vivo biodistribution of
nanocarriers over time. After administration, nanocarriers or their de-
gradable soft components would reach biological fluids and may in-
teract with these biological fluids (e.g. blood serum) or biomolecules
(e.g. proteins), which could significantly change their physicochemical
properties as well, such as size, drug loading, release profile and ag-
gregation or agglomeration, and alter the function of nanomedicine in
biological systems [50–53]. It is essential to fully characterize the na-
nomedicine products under clinically relevant conditions using in vitro
and in vivo models [54].
Although pictorial biodistribution of nanoparticle accumulation in
vivo can be obtained by fluorescence or radiolabelling method [55–57],
it is still very hard to quantify mass-balance information which is
paramountly important to account the full administered dose and en-
sure safety [58]. Usually the fluorescence and radiolabeling emitters
chemically conjugated to one composition of nanoparticles, which is
unstable and easily degraded in the internal environment. Therefore, it
may lead to unreliable results by tracing the degraded moiety of
fluoresce or radio molecule instead of the whole nanoparticle. Ad-
ditionally, the distribution of fluorescent or radio-modified nano-
particles may have different properties in comparison with the same
nanoparticles without fluorescence or radio-modification. Last but not
least, nanomedicine with more than one composition and the ability to
carry and deliver multiple therapeutic and imaging agents may require
individual tracking [59].
Consequently, multiple standard characterization methods in-
cluding the quantification of active and inactive ingredients and im-
purities, visualization of nanoparticles by microscopy, measurement of
particle size and size distribution with light scattering, and new ad-
vanced techniques specifically for the characterization of nanoparticle
in vivo behaviors are necessary to ensure that the nanomedicine pro-
ducts have all the desired properties for the intended therapeutic pur-
pose on a batch-to-batch basis with fewer side-effects [28].
2.3. Scale-up and manufacturing
The key issue for successful scale-up and manufacturing of nano-
medicine is controlling and keeping constant stability of physico-
chemical properties on a batch-to-batch basis [60], which is the most
challenging problem in pharmaceutical development.
Usually, preclinical and early clinical studies just need a small
amount of nanomedicine products. Small-scale processes may achieve
reproducibility with well-characterized nanoparticles. However, in
large-scale production, the polydispersity of the nanomaterial is diffi-
cult to control, which could cause batch-to-batch variations of physical
and chemical properties [61]. Since most nanoparticles are three-
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Materials Science & Engineering C 106 (2020) 110302
dimensional complex products with specific arrangement of multi-
component, which make the chemistry, manufacturing, and control
process very demanding. And traditional pharmaceutical manu-
facturing does not have rich experience to create nanoscale three-di-
mensional multicomponent systems. All of these make a series of blocks
for the scale-up and manufacturing of nanomedicine. An example is in
the production of nanomedical therapeutics Doxil®, the first FDA-ap-
proved nano-drug (1995) [7,62,63], which had to be suspended in
November 2011 because of its manufacturing and sterility issues. The
shortages of Doxil® were overcome until 2014, and an alternate man-
ufacturing approach for Doxil® was adopted resulting in increased
medication costs and delayed therapies for patients [28].
A well understanding and identifying the components and their
interactions in the early development help define the key characteristics
greatly, such as the critical steps and analytical criteria, to ensure re-
producibility of the product during the larger-scale manufacturing. In
general, there are two categories of nanoparticle preparation methods,
“top-down” and “bottom-up” approaches [64]. Top-down methods
manufacture nano-entities from larger ones, such as grinding of parti-
cles using the milling technique. In contrast, bottom-up methods ar-
range smaller components into functional super-assemblies, such as
polymerization of monomers or molecular self-assembly to arrange
single/multi-molecule components into a useful nanostructure by nat-
ural driving [65]. During the nanomedical formulation process, the
high-speed homogenization, sonication, milling, emulsification, cross-
linking, evaporation of organic solvents, centrifugation, filtration, and
lyophilization are always employed. It is very important to consider
which approaches and parameters are suitable to scale up at the lab- or
small scale stage and these crucial process conditions will benefit later
development process. These parameters may involve the ratio of na-
nomaterials, drugs and targeting moieties, the type of organic solvent
and emulsifier/stabilizer/crosslinker, the oil-to-water phase ratio,
temperature, pressure, and pH [65]. Choosing the wrong conditions
could lead to altering the chemical structure of the therapeutic re-
agents, other accessories, and unpredictable impurities. Particularly, it
is very easy to change chemical structure and conformation of macro-
molecules (protein, peptide) by denaturation, crosslinking, coagulation,
and degradation. Thus, the manufacturing of nanomedicine products is
not a simple addition and mixture of individual components, while
well-defined production steps, accurate analysis and strict quality
control are required [66].
The high costs of the raw materials and the need for a cockamamie
multistep production process are also obstacles for the scale-up and
manufacturing of nanomedicine, which makes the production of nano-
therapeutics very expensive. This may hinder pharmaceutical compa-
nies from carrying out the large-scale production of nanocarriers. For
instance, the expense of manufacturing commercial nano-formulation
drugs such as Abraxane® and Doxil® is far more than the cost of their
free drug paclitaxel and doxorubicin, receptively [67]. Therefore, in
order to compensate for the high costs on the development and man-
ufacture of nanomedicine products [68], the clinical therapeutic effect
of nanomedicine drugs has to be much more advanced than conven-
tional therapeutics.
2.4. Pharmacology and safety challenges
Physicochemical characteristics of nanomedicine notably influence
the pharmacological and safety profiles. Even subtle changes in com-
position and small deviations in the final manufactured products could
result in significant changes in pharmacology and toxicity of nanome-
dicine [28,66].
One of the requirements of a successful nanomedicine is to achieve
the desired pharmacological profile, for example pharmacokinetic
parameter (PK) [57]. However, most of the researcher and pharma-
ceutical companies apply the standard criteria of small molecule to the
assessment of nanomedicine. Usually, small molecular drugs alone
L.-P. Wu, et al.
diffuse more readily through biological barriers and the drug con-
centration in blood is somewhat in equilibrium and related to achiev-
able target tissue levels [28,66]. Thus, the measurement of drug con-
centration in the blood or plasma used as the standard approach to
determine PK is reasonable. But if apply this methodology to nano-
medicine, it cannot be assumed to be correct and could be inherently
flawed. Accordingly, different PK approaches with different indications
are required for different nanomedicine products. For example, instead
of the standard test of the behavior in plasma, the pharmacological
parameters at the specific target site could be more relevant to evaluate
the therapeutic activity of nanomedicine and the reproducibility. Fur-
thermore, the bioequivalency of nanotechnology-based products might
use these methods to evaluate as well.
In recent years, nanoparticles have been wide-spread used in med-
icine, which makes it necessary to address toxicity issues for human
health [69,70]. The nanoscale dimension of nanomedicine products can
simulate the intracellular organelles or biomolecules such as protein,
polysaccharide, and enzyme involved in cell signaling, which may lead
to detrimental biological interactions. Now the nanotoxicology is an
independent field of research [45,71]. And more and more data re-
garding the toxicity of different nanoparticles are becoming available.
Nonetheless, it remains difficult to evaluate the toxicity of nano-
materials, especially in vivo and long-term toxicity. At present, the as-
sessment of nanoparticle toxicity using classical drug toxicity assays
may be inadequate. And there is no standard list of required tests. Thus,
the development of advanced complementary assays and the standard
criterion for toxicity evaluation of nanomedicine products become
emergent [72,73]. There are multiple properties, such as size, shape,
surface area, surface charge, porosity, or hydrophobicity, which affect
the behavior and performance of nanomedicine drugs at the nano-bio
interface. Hence, each nanomedicine product may have its own case by
case issues and require particular evaluations. Usually, the toxicity in
pharmaceutical industry refers to acute toxicity and chronic toxicity.
Acute toxicity generally includes hemolysis, oxidative stress, in-
flammation, impaired mitochondrial function [74], or complement
activation [75–77]. Investigation of chronic toxicity is time-consuming,
and analyzing chronic toxicity data is more demanding.
Due to the lack of suitable animal model, immunotoxicity cannot
readily carry over from preclinical testing to humans. Nanoparticles
themselves and the biologics such as proteins, peptides, antibody
fragments, and nucleic acids in nanoparticles can serve as antigens
sources that can elicit the immune response. The immunogenicity of
nanoparticles can be affected by their physicochemical properties, such
as size, surface characteristics, charge, hydrophobicity, and solubility.
2.5. Regulatory challenges
Although the FDA and European Medicines Agency (EMA) approved
a number of nanomedicine products for cancer therapy, there are not
specifically implemented guidelines for drug products containing soft
materials by FDA, EMA, and other regulatory bodies yet. The lack of
guidance in the examination of nanomedicine, nanomedicine ther-
apeutics can only make regulatory decisions based on individual as-
sessment of benefits and risks [66]. In this way, the regulatory process
is time-consuming and requires the high-level expert in innovative
technologies, which may result in regulatory delays. And regulatory
issues are important for the development of cutting-edge technologies
to characterize and monitor the quality of nanomedicine products be-
side clinical trials and the approval process. Especially there is an ur-
gent need for detailed guidelines for characterization and quality con-
trol of nanomedicine products [78]. The good thing is that the
definitions, guidelines, and cooperation in this area are gradually es-
tablished and improved [79–81]. For example, the guidance for in-
dustry entitled “Considering Whether an FDA-Regulated Product In-
volves the Application of Nanotechnology” is released by the FDA in
June 2014 [82], where the definition of nanomaterials is engineered
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Materials Science & Engineering C 106 (2020) 110302
materials with at least one dimension between 1 and 100 nm. Also, the
FDA and European Technology Platform on Nanomedicine (ETPN) in-
tends to collaborate with the Nanotechnology Characterization La-
boratory (NCL) and European Nano-Characterization Laboratory (EU-
NCL) respectively to promote the regulatory review and in-depth
characterization of nanomedicine products [83]. The urgent demand
for regulatory agencies in nanomedicine therapeutics is to refine and
standardize requirements for the approval of safe nanomedicine pro-
ducts. Also, more advanced and multifunctional tools need to be de-
veloped, even though the complexity of the approval process could be
increased.
3. Solution and conclusions
In summary, nanoparticles have made major contributions to clin-
ical medicine in the areas of therapeutics and diagnosis, and several
innovations for example stealth and long circulating liposome are by
now well-established and transfer into clinical applications, newer
technologies continue to emerge following the same basic concepts of
design. But from the therapeutic angle, nanomedicine is still in infancy
and need to overcome several hindrances before it is approved for
marketing [6]. Firstly, it needs to design and engineer nanostructures
with appropriate physicochemical properties and biological behaviors,
optimize reproducible manufacturing processes, select reliable analysis
methods for adequate characterization. For example in the size esti-
mation, dynamic light scattering (DLS) is widely used for the size
analysis of nanoparticles in liquid solution [84]. The average particle
size from DLS analysis is heavily biased towards small numbers of large
particles because the intensity of light scattered from a nanoparticle
varies as a function of radius to the power of six for a Rayleigh particle
[84]. NTA (nanoparticle tracking analysis) is an alternative modality to
DLS, which is gaining popularity [85]. Unlike DLS, NTA does not use
the intensity of light scattered as a measure, but measures the particle
size from the changing position of scattering diffusing particles through
image analysis. This process overcomes inherent problems with DLS
and can be used for monitoring polydisperse samples as well as ag-
gregates [85]. Fig. 5 shows a representative example of NTA analysis,
demonstrating population changes that have occurred on coating a
polymeric nanoparticle suspension with polyethylenimine [26]. Also,
the nanomedicine products should have favorable pharmacology and
toxicity profile, and demonstrate safe and efficiency in clinical trials. To
make this happen and get the attraction for pharma development,
collaboration is a key factor to the future success of nanomedicines. The
development of nanomedicine is a multidisciplinary endeavor, and it is
necessitates highly varied expertise to develop translatable therapeutics
because of the diversity and complexity. It is essential to the successful
cooperation from the experts of biology, chemistry, nanotechnology,
imaging, and medicine. It is important to foster partnerships between
large pharma, smaller companies, and academia early in preclinical
development to capitalise on the unique strengths of each partner. Ef-
fective integration of resources from academia, industry, consortia, and
hospitals will be essential to help the link of physico-chemical proper-
ties of nanomedicine systems with biological implications. Finally,
health authorities should develop and consummate definitions, quality
standards, guidelines and regulatory issues of nanomedicine. Although
any new drug has to face similar hurdles conceptually, the particular
complexity and multicomponent nature of nanomedicine lead to a large
number of additional variables that may substantially increase more
considerations in controlling processes and monitoring of behavior in a
biological system. Through rational designed and systematic ap-
proaches to meet these challenges and solve the problems, nanomedi-
cine will forward to a new step and provide realistic and significant
value to human medicine and healthcare.
L.-P. Wu, et al.
Fig. 5. Characteristics of engineered PHBHHx nanoparticles before and after PEI coating. Typical size distribution profile of PHBHHx nanoparticles before (A) and
after PEI coating (B) as a function of particle concentration determined by nanoparticle tracking analysis. The insets represent an example of the real-time capture
video frame of point scatters. (C, D) 2D plots of relative light scattering intensity of particles versus the estimate of the particle size (Reprint from Ref. [26]).
Declaration of competing interestCOI
The authors declare that they have no conflict of interest.
Acknowledgement
L-P.Wu. acknowledges financial support from National Science and
Technology Major Projects for Stem Cell and Translation (reference
2019YFA0110500), National Science and Technology Major Projects
for New Drug Development (reference 2018ZX09733-006), Guangdong
Pearl River Talents Program (reference 2017GC010411), International
Science and Technology Cooperation Project of Guangzhou Economic
and Technological Development District (reference 2018GH15) and
Drug Discovery Pipeline of Guangzhou Institutes of Biomedicine and
Health (reference 201508020131).
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