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372 D. R. Serrano Lopez et al.
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Fig. 14.8 Top-down manufacturing of nanomedicine
purpose of the stabilizing excipient is to prevent occulation and crystallization, known as Ostwald ripening (Ostwald 1900). If the obtained suspension is chemically and physically stable, it can be preserved in its current state. However, if the suspension is not stable, additional processing is required for which spray drying, freeze drying (Müller et al. 2001), or uid bed granulation can be used (Dua et al.
2012; Serrano et al. 2018) (Fig. 14.8
In comparison to the bottom-up method, the top-down method offers numerous advantages. These are (i) no organic solvent, (ii) poorly soluble drugs can be used, (iii) less labour-intensive work, (iv) lower risk of polymorphic transitions (Sharma et al. 2009), (v) possibility to process highly concentrated suspensions, (vi) the scaling up process is feasible, and (vii) target particle size easier to obtain (Cornier et al. 2017).
).
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14.6.2 Novel Approaches in the Continuous Manufacturing
of Targeted Nanomedicines: Microuidic Chips
The conventional batch manufacturing of nanomedicines is shifting towards contin­uous manuf acturing in which raw materials are continually injected into a manufacturing facility, and products are continuously discharged during the opera­tion of the manufacturing processes (Inada manufacturing requires a fully automated system and continuous monitoring in real time (Bohr et al. time screening. The microuidic device refers to a component that has microuidic channels handling very small uid volumes (Song et al. manufactured using polymer materials such as polymethylmethacrylate, polysty­rene, polycarbonate, and polydimethylsiloxane (PDMS), being the latter the most frequently used material (Wu et al. 3D printed microuidic chips are also a novel alternative to PDMS chips being able to personalize the design of microuidic chips adapted to the nanomedicine features (Kara et al.
Commonly, chips have two or three liquid entrance ports and one exit port. Similar to conventional methods, the drug, and the polymeric or lipidic excipients solubilized in a water-miscible solvent will pass through one of the inlet ports while water containing a surfactant will run through the other port (Fig. 14.9). The uid ow is controlled by the same rules governing the ow of a uid at the macroscale. Microuidic devices are not simply a miniaturized type of their macroscale versions, due to several physical features (e.g. high ratio of surface/volume and mass transfer based on diffusion) that do not linearly scale from macrodomains to microdomains. The key points to select before manufacturing nanomedicines are the total ow rate, the ratio between the aqueous and the solvent phase, and the composition of the particles. The self-assembly occurs inside the micrometric channels. When param­eters are properly selected, high encapsulation loadings can be achieved (<90%) with tuneable particle size. Solvent removal takes place by dialysis (in batch manufacturing) or using ow-tangential units (Osouli-Bostanabad et al. 2022). Currently, a few industrial companies, such as Precision Nanosystems, have devel­oped microuidic units allowing the manufacturing of 10 L batch size continuously.
2019). Microuidic devices allow for continuous ow with real-
2003; Xia et al. 1996; Xia and Whitesides 1998).
2021).
2020; Nambiar et al. 2022). Continuous
2018) and can be
14.7 Conclusions and Future Perspectives
To date, a few passive targeted nanomedicines have been commercialized (Osouli­Bostanabad et al. 2022; Lammers et al. 2012). Although signicant progress has been made in the eld of actively targeted nanomedicines, the clinical translation of promising approaches from bench to bedside has been poor at best (Bae and Park
2011b). Many times the potential usefulness of active targeting is exaggerated,
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Fig. 14.9 Continuous manufacturing of nanomedicines using microuidic chips. (Reproduced from Osouli-Bostanabad et al.
2022)
especially in cancer therapy. There are several important pitfalls that hamper the clinical success of these nanomedicines such as:
(i) Overinterpretation of the EPR and ELVIS effect. Although the EPR and ELVIS
effect enhances the accumulation of nanomedicines at the intended site, this does not mean that 100% of the dose administered will be accumulated at the target site. Actually, in the majority of the intravenous administered formula­tions, above 95% o f the dose ends up at unintended sites (Bae and Park
2011b).
(ii) Poor tumour penetration. Even if the nanomedicine has a suitable size, shape,
and surface charge to be extravasated through the opening between endothelial vascular cells within the tumour tissue, it should not be forgotten that the higher interstitial uid pressure (ranging from 5 to 40 mm Hg) inside most solid tumours compared to normal tissues (typically <3 mm Hg) will make accu­mulation and diffusion of the nanomedicine within the tumour tissue difcult (Milosevic et al. 2004).
(iii) Overexpression of the target and tumour heterogenicity. There are two major
problems regarding these concepts. First, it is very difcult to nd a target that is exclusively expressed by cancer cells (or the blood-brain barrier), and second, the target is not usually expressed by all the cancer cells. Even though certain receptors could be over-expressed on cancer cells, one should consider how many times the mass of non-target cells is larger than the target cell mass (Bae and Park 2011b).
Overcoming the drawbacks regarding the formulation design such as solubility, aggregation, instability, suitable particle size, shape, and surface can result in too complex nanomedicines and low-cost effectiveness which make them not widely
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Fig. 14.10 Requirements for optimal active targeted nanomedicines and problems to be addressed
applicable. Besides difculties in formulation design and manufacture, once nanomedicine have been administered, they will face both anatomical and physical barriers that have to be overcome to elicit a pharmacological effect (Fig.
14.10)
(Lammers et al. 2012). For these reasons, it is essential to develop appropriate animal models able to reproduce all the hurdles mentioned above and predict better the clinical situation. Also, the development of continuous manufacturing methods, such as microuidic chips, is key to ensuring a faster clinical translation with lower batch­to-batch variability.
Finally, future active targeting nanomedicines could yield multifunctional per­sonalized theranostic therapies which will combine therapeutic and diagnostic functionalities.
Nanomedicines have to overcome a number of design hurdles to achieve targeting, such as surface hydrophilicity, charge, particle size and shape. Upon intravenous administration, nanomedicines have to avoid the opsonization by plasma proteins to avoid being phagocyted by the macrophages of the reticuloen­dothelial system. Also, nanomedicines should exhibit sufcient residence in the bloodstream. Because of the larger blood vessel fenestrations in the liver (100–175 nm), particles can easily be extravasated in the liver and metabolized (Ballet weight <40,000 Da will be cleared by the kidneys (Yokoyama
1990). In addition, nanomedicines with a diameter <10 nm or molecular
2005). In the case of
cancer active targeting, the EPR effect plays a key role by enhancing the retention of
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the nanomedicine within the tumour tissue. The pore cut-off size of the blood vessels in the majority of the tumours ranges from 380 to 780 nm unlike the small pores (around 4.5 nm) present in normal tissue endothelium (Rippe et al. 2002; Ballet
1990). Once the nanomedicine has accumulated in the tumour, specic interactions
between the targeting moieties and the tissue receptors need to occur. Finally the therapeutic and/or imaging agent needs to be released. Additionally, the nanomedicines will have to overcome the high interstitial uid pressure and poor penetrability in the tumour tissue as well as avoid that their contents are expelled by the drug ef ux pumps.
Problems
Question 1 Apart from the hydrodynamic size and the shape, which other characteristic is important to bear in mind in the choice of nanocarrier?
Surface properties such as charge and hydrophobicity play a key role in the cellular uptake, clearance, and biodistribution of nanomedicines. For example, hydrophobic and charged nanocarriers have shorter blood circulation times because of the adsorp­tion of the opsonin proteins onto their surface that render them easily recognizable by macrophages of the reticuloendothelial system and thus, they will not arrive at the desired target to elicit their effect.
Question 2 What are the major advantages of using antibodies fragments as targeting moieties compared to the whole antibody? Antibody fragments
are smaller in size and lack the Fc domain and the complement-activating region (which might reduce the immune response) while keeping their antigen-binding afnity. In addition, their identication and manufacture are easier compared to whole antibodies.
Question 3 Are there any continuous manufacturing methods to fabricate nanomedicines? Yes, microuidic chips are a promising tool able to manufacture
nanoparticles with tuneable physiochemical characteristics.
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