HK1191553B - Systems and methods for producing multilayered particles, fibers and sprays and methods for administering the same - Google Patents

Description

System and method for producing multilayer particles, fibers and sprays and method for applying same

This application is a divisional application of the patent application having application number 200780023710.4, application date 2007, 5/3, entitled "System and method for producing multilayer particles, fibers and spray and method for applying same".

Cross Reference to Related Applications

Priority and benefit of the U.S. provisional application serial No. 60/746,311 filed on 3.5.2006 and 60/886,225 filed on 23.1.2007 are claimed herein, the disclosures of which are expressly incorporated herein in their entirety by reference.

Technical Field

The present invention relates to systems and methods for producing capsules and particles containing at least one encapsulated and/or embedded agent, such as therapeutic agents, imaging agents, and other ingredients. More particularly, the agent encapsulated in a medium, capsule, particle, mediator, or carrier may maximize treatment and/or imaging of malignant tumors while minimizing adverse effects of the treatment and/or imaging.

Background

Cancers are a class of diseases or disorders characterized by uncontrolled cell division and their ability to spread by invasive growth directly to adjacent tissues or by metastatic implantation to distant locations. Cancer can affect people of all ages, but the risk tends to increase with age. Cancer is one of the leading causes of death in developed countries.

For example, in 2003, 12 to 13 out of 10000 women suffered from breast Cancer, according to the National Cancer Institute. Although imaging and early diagnosis tools have been improving over the past two decades, early detection of breast cancer is far from being absolutely reliable, especially when mammograms are used in young women. While magnetic resonance imaging (MRI and ultrasound) and laser-based imaging techniques have been used or evaluated, the problem of the best possible contrast between healthy and cancerous tissue is a long standing problem for any imaging technique.

The challenge facing scientists today is to determine how to design therapeutic or imaging agents and their mediators, mediators or carriers in order to maximize the treatment and imaging of malignant cancers in patients while minimizing the adverse effects of the treatment. In addition, selective delivery of therapeutic agents to desired parts of the body is also a matter of great concern. Current treatments may result in inadequate tumor distribution of the therapeutic agent and often cause adverse effects to the patient. Systemic injection of therapeutic agents results in consequences associated with their non-specific dispersion within the body, and there is limited distribution of therapeutic agents throughout the targeted malignancy. One approach to overcoming these disadvantages is to design an effective therapeutic or imaging agent delivery vehicle by preparing vesicles or capsules containing the desired therapeutic agent.

The formation of vesicles or capsules, which are small enough to be delivered into the human body by inhalation, injection or permeation through the skin, has received significant attention. The outer epidermis or shell of the vesicle may be chemically functionalized with receptors and other species to selectively target certain organs. There are several methods available to make vesicles, such as electrospray and dual-capillary jet systems.

Emulsion-polymerization techniques such as DC coaxial electrospray, AC coaxial electrospray, and Electrohydrodynamics (EHD) are well known methods by which micron-sized capsules can be produced. Typically, a solution containing the compound of interest to be encapsulated (also referred to as an "encapsulate") is emulsified into another fluid as a solution having at least one substance capable of forming a shell or coating around the dispersed droplets of the encapsulate. Although the emulsion-polymerization process is relatively scalable, there are several limitations associated with this process, such as the inability to encapsulate the target in a quantitative manner, and the high shear production of emulsions can compromise the integrity of mechanically delicate encapsulates (e.g., biological components such as proteins, genetic material, and other biologically derived molecules).

Coaxial electrospray (which is based on the application of a DC potential between a coaxial capillary device delivering a core fluid containing encapsulates and a shell-forming fluid precursor and a collector surface or counter electrode) has the ability to generate vesicles in the micron and submicron range. Although DC coaxial electrospray may be relatively gentle for biological encapsulates, its main limitation is its lack of simplicity in equipment, design, scalability, and cost effectiveness.

AC electrospray may be used as an alternative to DC-based electrospray. For example, AC electrospray can be used to produce encapsulates embedded in a bioabsorbable biopolymer matrix, such as polylactic acid. AC electrospray produces essentially electrically neutral electrospray. Despite the advantages associated with charge neutrality, such as reduced ability of the particles or electrosprays to differentially adsorb on non-targeted surfaces, and avoidance of potential charge accumulation problems. One disadvantage of AC electrospray is that it produces undesirably large particles having a size much larger than 1 micron. For many medical applications, such as crossing the blood brain barrier, AC electrospray produces particles that are unacceptably large.

In addition to electrospray, coaxial fluid jet systems such as EHD combined with sol-gel chemistry can also be used to make vesicles or capsules. It is known in the art to use a coaxial, double-capillary coaxial arrangement to deliver two fluids simultaneously in the presence of an electric field gradient.

In short, in this method, the chemical and physical properties of the two fluids and the values of the variables, such as the electric field strength and the flow rate of the two fluids, determine the structure of the material collected on the collector electrode, which may be located away from the outlet region of the double-capillary coaxial arrangement. At the exit region, the compound bi-fluid structure may form a charged meniscus that may take a variety of shapes, such as a Taylor (Taylor) cone.

Fig. 1, which illustrates fluid flow produced by a prior art dual-coaxial capillary system, depicts a dual-fluid flow 100 in the presence of an electric field gradient, in which an inner fluid 104 is encapsulated by an outer fluid 106. The quasi-conical taylor cone structure 108 emits an electrically charged composite bi-fluid jet from its tip 110. Here, the charged fluid jet undergoes refinement due to the same charge repulsion. Furthermore, the refinement of the jet may be a function of two fluid physical properties, such as dielectric constant, viscosity, conductivity, and surface tension. Although differentiated bi-fluid structures may occur when the fluid 104 and the fluid 106 are not mixed, they may also occur when the fluid 104 and the fluid 106 are miscible or partially miscible, since both fluid flows are under the so-called laminar flow regime.

Laminar flow may be non-turbulent, which may minimize mixing between layers of flowing fluid. Thus, the refined charged bi-fluid jet may enter chaotic channels similar to the phenomenon of oscillation, since the two fluids are not mixed to the extent that a single fluid phase is formed. At some point along its path to the collection region or collector body 112, the composite bi-fluid jet may experience a charge oscillation phenomenon known as Rayleigh instability. This may cause the composite bi-fluid jet to no longer undergo progressive refinement, but to undergo an oscillating refinement and thickening regime that may ultimately lead to a regime in which the jet breaks up into droplet-in-droplet droplets, or a composite electrospray regime.

The chemical and physical properties of the two fluids may be controlled to produce a variety of structures that collect at the collection region or collector 112. For example, if the fluid 106 produces a solid structure by solvent evaporation and solid phase precipitation, the fluid 104 may be encapsulated into a structure such as a hollow fiber, hollow bead fiber, or capsule, for example. Alternatively, during the time that the composite charged structure travels from the dual-fluid charged meniscus to the collection region 112, the chemical and physical properties of the two fluids may be adjusted so that there is no curing of the fluids 104 and 106, one of the fluids is cured, or both fluids are cured.

With respect to fig. 1, regions 3,4 and 5 are shown. If certain physicochemical phenomena cause the fluid 106 to solidify in region 3, it is possible to obtain a tubular structure that encapsulates the fluid 104 in a solid shell formed by the fluid 106. However, if the solidification phenomenon in the fluid 106 occurs in region 4, it is possible to obtain hollow bead fibers containing the encapsulating fluid 104. Alternatively, if a solidification phenomenon in the fluid 106 occurs in the region 5, it is possible to obtain a capsule containing the encapsulated fluid 104. With the fluid 104 solidified but the fluid 106 not, wetted fibers, wetted beaded fibers, or wetted particles may be produced in regions 3,4, and 5, respectively. When the two fluids solidify before reaching the collection area, a core-shell solid structure may result.

While coaxial double-capillary systems can be used to create the core-shell structure described above, there are several disadvantages associated with this conventional system. In particular, micro-fabrication problems can occur when attempting to scale the process directly, in parallel, to increase process throughput. For example, typical ranges for the inner diameters of the inner and outer capillaries are about 0.1 to about 0.3mm and about 0.3 to about 1.0mm, respectively. In order to build up an apparatus consisting of a number of such coaxial double-capillary devices for the production of the desired core-shell structure on a large scale, it is necessary to produce individual devices having inner and outer capillaries arranged as close to coaxial as possible and also having a high degree of reproducibility in their diameter. Modern microfabrication techniques, however, can encounter challenges that are very complex and not cost effective.

In particular, the conventional way of handling fluid flow through a number of orifices, capillaries, conduits or double-capillary co-axial devices is by using one device that passes fluid flow through all of the same fluid orifices, capillaries, conduits or double-capillary co-axial devices, rather than by controlling the fluid flow rate through each individual fluid flow path. This is why it is difficult and expensive to produce the desired core-shell structure on a parallel scale, for example. If the diameter variation from the inner or outer capillary of one dual-capillary coaxial device to the inner or outer capillary of the other is more than about 2% or 3%, it is not possible to produce the desired core-shell structure without undesirable structures. With this prior art dual-capillary device, small differences in the overall pressure differential pattern of the inner and outer capillaries can also cause undesirable effects.

Disclosure of Invention

SUMMARY

The present invention meets the above needs by providing systems and methods for preparing capsules with embedded or encapsulated therapeutic and/or imaging agents. More particularly, the system and method of the present invention allows for a wider range of capsule sizes, maintains chemical interactions that result in degradation of the therapeutic and/or imaging agents to sufficiently low and acceptable levels, includes fewer preparation steps, and is more cost effective than conventional methods.

According to an aspect of the present invention, a method of producing a capsule containing at least one encapsulated agent is provided. The method may comprise providing a core fluid to the interior of the hollow tube, providing a shell fluid stream, generating a shell fluid stream at the outer wall of the hollow tube, subjecting the core fluid and the shell fluid to an electrical potential so as to form a bi-fluid charged jet at the top of the hollow tube, and forming a capsule having a core region and a shell region at the top of the hollow tube. Furthermore, the method may further comprise providing a current collector and depositing the formed capsules on a surface of the current collector.

In another aspect, the method of the invention may include dispersing at least one magnetite particle within the shell region of the capsule. The at least one magnetite particle may have a size in the range of about 1nm to about 300 nm. The magnetite particle species may be Fe₃O₄。

In yet another aspect, the method can include dispersing at least one photon-sensitive nanoparticle within a shell region of the capsule. The photosensitive nanoparticle species may have a size in the range of about 1nm to about 50 nm. This class of photon-sensitive nanoparticles may include silver, gold, palladium, and any combination thereof.

In a further aspect, the core fluid may be provided at a flow rate in a range of about 0.005 ml/hr to about 5 ml/hr, and in particular, at a flow rate of about 0.025 ml/hr. The shell fluid may be provided at a flow rate in the range of about 0.005 ml/hr to about 5 ml/hr, and in particular, at a flow rate of about 0.75 ml/hr. The positive electrical bias may have a voltage of about 5kV to about 18 kV. The collector may be at ground potential.

In one aspect, the method can include dispersing functional groups at the periphery of the capsule shell. The functional group can be an entity such as a hydroxyl group, an amino group, a carboxyl group, a carboxylic anhydride group, a mercapto group, a hydrosilyl group, a thio group, a carboxylic acid group, an amine group, and any combination thereof. In addition, the functional group may be capable of chemically or physically associating with pancreatic cancer cells. The functional group can be one or more compounds such as kinase receptors, fibroblast growth factor receptors, EGF, TGF, VEGF-A, urokinase receptors, interleukin-4 receptors, retinoic acid receptors, heparin-binding EGF-like growth factor, HB-EGF, amphiregulin, epiregulin, neuregulin, and functional equivalents thereof. The functional group may have a content ranging from about 0wt% to about 1 wt%. In addition, the functional group may be capable of binding subcellular targets such as endoplasmic reticulum, mitochondria, golgi apparatus, vacuoles, nuclei, acrosomes (aerosomes), centrosomes, cilia, glyoxylate cycle bodies, lysosomes, melanosomes, myofibrils, nucleoli, peroxisomes, actin, tubulin, plasma membranes and ribosomes, vesicles.

In a particular aspect, the functional group may be capable of chemically or physically attaching to glioma cells and may be epidermal growth factor and functional equivalents thereof. The functional group may have a content on the capsule in a range of about 0wt% to about 0.02 wt%.

In another particular aspect, the functional group can be capable of chemically or physically associating with attachment to a breast cancer cell. The functional group may be estrogen and functional equivalents thereof. Additionally, the functional group may be present on the capsule in an amount ranging from about 0.02wt% to about 0.4 wt%.

In yet another particular aspect, the functional group is capable of chemically or physically associating with at least one of lymphoma, myeloma, and leukemia cancer cells. The functional group may include one or more of the following: VEGR-2, tositumomab (tostmomab), antibodies capable of binding to the CD20 receptor, CD5, CD7, CD13, CD19, CD22, CD33, CD52, CD61, anti-myeloperoxidase and functional equivalents thereof. The functional group may have a content on the shell of the capsule in the range of about 0.0wt% to about 1.0 wt%.

According to an aspect of the invention, the shell fluid may be at least one biocompatible polymer. The biocompatible polymer may include one or more of the following: poly (lactic acid), poly (lactic-co-glycolic acid), chitosan (cetosan), alkyl methacrylates, polycaprolactone, starch, polyethylene glycol, and copolymers resulting from combinations thereof.

In particular aspects, the core fluid may include one or more of: therapeutic agents and imaging agents. The therapeutic agent may include one or more materials such as nucleic acids, double-stranded DNA, single-stranded DNA, double-stranded RNA, single-stranded RNA, Peptide Nucleic Acids (PNA), antisense DNA, antisense RNA, small interfering RNA, proteins, lipids, carbohydrates, and combinations thereof. Further, the therapeutic agent is at least one non-biological material. The imaging agent is suitable for detecting malignant cancer in a patient.

In another aspect, the therapeutic agent may be useful for treating a patient suffering from a disorder. The patient may be afflicted with at least one disorder such as breast cancer, glioma, lymphoma, leukemia, prostate cancer, pancreatic cancer, carcinoma, sarcoma, mesothelioma, glioma, germ cell tumor, and choriocarcinoma.

According to an aspect of the invention, an electrohydrodynamic system for producing a capsule having at least one encapsulated agent is provided. The system may include a hollow tube having an interior configured to receive a core fluid, a fluid source surrounding the hollow tube, a core fluid supply tube arranged to supply the core fluid to the interior of the hollow tube, a shell fluid supply tube arranged to supply the shell fluid to the shell fluid source, and an electrical potential source that subjects the core fluid and the shell fluid to an electrical potential to cause the fluids to form a jet comprising at least a bi-fluid electrically charged fluid. The system may include a wick fluid reservoir. The system may also include a shell fluid reservoir. The encapsulated agent may be at least one of a therapeutic agent and an imaging agent.

In particular aspects, the system can further include a collector electrode positioned above the fluid slot. Additionally, the system may include an extractor body positioned between the fluid bath and the collector electrode.

In a more particular aspect, the fluid source can be a fluid sink. The fluid source may also be a porous material. The porous material may be a sponge. Further, the fluid source may be a plurality of tubes.

According to an aspect of the invention, a system for producing a capsule with at least one encapsulated medicament is provided. The system may comprise a plurality of hollow tubes, a jacket surrounding the plurality of hollow tubes, a core fluid supply tube arranged to supply the core fluid to the interior of the plurality of hollow tubes, a shell fluid supply tube arranged to supply the shell fluid, and a potential source that subjects the core fluid and the shell fluid to a potential such that the streaming fluids form a jet having at least a bi-fluid electrically charged fluid. The system may also include a collector electrode. In addition, the system may further include an extractor body.

In a further aspect, the system of the present invention may include a wick fluid reservoir. In addition, the system may further include a shell fluid reservoir.

In a particular aspect, the plurality of hollow tubes may be arranged circumferentially within the sleeve. The plurality of hollow tubes may also be linearly arranged in said sleeve, which sleeve comprises at least two plates. The sleeve may be configured to receive and deliver the shell fluid. The shell supply tube may be configured and arranged to supply the shell fluid to a space between the plurality of hollow tubes.

In particular aspects, the system of the present invention can be assembled to achieve up-flow hydrodynamics. Alternatively, the system of the present invention may be assembled to achieve downstream electrohydrodynamic.

Other features, benefits and embodiments of the present invention can be described or made apparent from consideration of the following detailed description, drawings and claims. Furthermore, it is to be understood that both the foregoing summary of the invention and the following detailed description are exemplary and intended to provide further explanation without limiting the scope of the invention as claimed.

Detailed Description

It is to be understood that this invention is not limited to the particular methodology, protocols, reagents, etc. described herein as these may vary, as will be appreciated by one skilled in the art. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention. It is also noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Herein, for example, reference to "a capsule" is a reference to one or more capsules and their equivalents known to those skilled in the art.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The details of the embodiments of the invention and their various features and benefits are explained more fully with reference to the non-limiting embodiments and/or are illustrated in the accompanying drawings and detailed in the following description. It should be noted that the features illustrated in the drawings are not necessarily drawn to scale and features of one embodiment may be employed with other embodiments, as the skilled artisan will appreciate, even if not explicitly stated herein.

Any numerical value recited herein includes all values from the lower value in increments of one unit to the upper value, provided that there is a separation of at least 2 units between any lower value and any higher value. For example, if it is stated that the concentration of a component or a value of a process variable, such as size, temperature, pressure, time, etc., is, for example, from 1 to 90, particularly from 20 to 80, more particularly from 30 to 70, it is intended that values such as 15 to 85, 22 to 68, 43 to 51, 30 to 32, etc., are expressly enumerated in this specification. For values below 1, suitably 1 unit is considered to be 0.0001, 0.001, 0.01 or 0.1. These are only examples of what is specifically mentioned and all possible combinations of numerical values between the lowest value and the highest value enumerated are to be considered to be expressly stated in this application in a similar manner.

Also provided immediately below is a "definitions" section, wherein certain terms that are pertinent to the present invention are specifically defined. Specific methods, devices, and materials are described, although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All references mentioned herein are incorporated by reference in their entirety.

Definition of

BSA is bovine serum albumin

CED is enhanced convective delivery

CFM is confocal fluorescence microscopy

EHD is electrohydrodynamics

PBS is phosphate buffer

PEG is poly (ethylene glycol)

PLA is poly (lactic acid)

PCL is polycaprolactone

PLGA is poly (lactic-co-glycolic acid)

The terms "active agent", "drug", "therapeutic agent" and "pharmacologically active agent" are used interchangeably herein to refer to a chemical substance or compound that induces a desired pharmacological effect when administered to an organism (human or animal). Included are derivatives and analogs of these compounds, or compounds of the classes specifically mentioned, which also induce the desired pharmacological effect. In particular, the therapeutic agent may comprise a single biological or non-biological compound, or a combination of biological and non-biological compounds, which may be desirable to elicit the desired therapeutic effect.

By "pharmaceutically acceptable carrier" is meant a substance or material that is suitable for pharmaceutical administration and is not biologically or otherwise undesirable, i.e., that can be administered to an individual along with an active agent without causing any undesirable biological effects, or that interacts in a deleterious manner with any of the other components of the formulation in which it is contained.

Similarly, as provided herein, a "pharmacologically acceptable" salt, ester, or other derivative of an active agent is not a biologically or otherwise undesirable salt, ester, or other derivative.

As provided herein, the term "effective amount" or "therapeutically effective" of a pharmaceutical agent means a non-toxic but sufficient amount of the agent to provide the desired therapeutic effect. The exact amount required will vary from subject to subject, depending on the age, weight and general condition of the subject, the severity of the condition being treated, the judgment of the clinician, and the like. Therefore, it is not possible to specify an exact "effective amount". However, in any individual case, an appropriate "effective" amount can be determined by one of ordinary skill in the art using only routine experimentation.

The terms "treatment" and "treating" as used herein refer to a reduction in the severity and/or frequency of symptoms, elimination of symptoms and/or underlying causes, the appearance of symptoms and/or prevention of their underlying causes, and the amelioration or healing of injury. Thus, for example, the method of "treating" a cancer individual, as the term "treatment" is used herein, includes the treatment of cancer in a clinically symptomatic individual.

The terms "disorder," "disease," and "disorder" are used interchangeably herein and refer to a physiological condition that can be detected, prevented, or treated by administration of a therapeutic agent as described herein. Exemplary diseases and disorders can include, but are not limited to, cancers such as breast cancer, glioma, pancreatic cancer, leukemia, and lymphoma.

The term "patient" in the treatment of "a patient" refers to a mammalian subject, e.g., suffering from or susceptible to a condition, disease or disorder as specified herein, and includes humans and animals.

The term "nucleotide" as used herein may include oligonucleotides, nucleotides or polynucleotides, and fragments thereof, relating to DNA or RNA of genetic or synthetic origin which may be single-or double-stranded; and denotes the sense or antisense strand, relating to Peptide Nucleic Acids (PNAs), small interfering RNA (sirna) molecules, any DNA-like or RNA-like material, of natural or synthetic origin.

The term "transfection" as used herein includes the process of inducing a DNA expression vector into a cell. Various methods of transfection are possible, including microinjection or lipofection.

The term "transformation" as used herein generally refers to a process by which exogenous DNA enters and alters a recipient cell. Which may occur under natural or artificial conditions using various methods well known in the art. Transformation may rely on any known method for inserting foreign nucleotide sequences into prokaryotic or eukaryotic host cells. The method is selected according to the type of host cell to be transformed, and may include, but is not limited to, viral infection, electroporation, heat shock, and lipofection.

Antisense gene: an antisense gene consists of reversing the orientation of the gene with respect to its promoter so that the antisense strand is transcribed.

Antisense RNA: an RNA molecule attached to a specific RNA transcript that can hybridize to the transcript and block its function.

The term "functional equivalent" as used herein generally refers to a protein or nucleotide molecule having functional or structural characteristics substantially similar to a heterologous protein, polypeptide, enzyme, or nucleotide. Functional equivalents of a protein may contain variants, depending on the requirements of the variant for performing a particular function. The term "functional equivalent" is intended to include "fragments", "mutants", "hybrids", "variants", "analogues" or "chemical derivatives" of a molecule.

"composite charged fluid jet" as used herein generally refers to a stream of fluid consisting of at least two distinct fluid layers. For example, when two fluids are used to form a composite charged jet by EHD, the outer fluid is a fluid precursor to the shell of a capsule prepared by the composite charged fluid jet EHD process; and the internal fluid is a precursor to the capsule core.

According to the invention, the medicament is "encapsulated" when the capsule has at least one core region and at least one particle shell region in a layered structure. The capsule may further contain at least one pharmaceutical agent, such as a therapeutic agent or an imaging agent.

According to the invention, the agent is "embedded" when said agent is dispersed in a biocompatible matrix consisting of one or more biocompatible polymers and the different onion-like layers are not evident.

The term "carcinoma" as used herein generally refers to a malignant tumor derived from epithelial cells. This group may represent the most common cancers, including common forms of breast, prostate, lung and colon cancer.

The terms "lymphoma" and "leukemia" as used herein generally refer to malignant tumors derived from blood and bone marrow cells.

The term "sarcoma" as used herein generally refers to malignant tumors derived from connective tissue or mesenchymal cells.

The term "mesothelioma" as used herein generally refers to a tumor derived from mesothelial cells lining the peritoneum and pleura.

The term "glioma" as used herein generally refers to the most common type of tumor derived from glia, i.e., brain cells.

The term "germ cell tumor" as used herein generally refers to tumors derived from germ cells, which are typically found in the testis and ovary.

The term "choriocarcinoma" as used herein generally refers to a malignant tumor derived from the placenta.

The term "attached" as used herein generally refers to covalent binding, adsorption, and physical immobilization. The terms "associated," coupled, "and" connected "equally mean the term" attached.

The term "nanoparticle" as used herein generally refers to a particle, typically a metallic, semiconducting, magnetic, ceramic, and insulating particle, having a diameter in the range of about 1nm to about 1000 nm.

The term "polymer" or "biopolymer" as used herein generally refers to compounds having two or more monomeric units and is intended to include linear and branched polymers and copolymers, and the term "branched polymer" includes simple branched structures as well as highly branched and dendritic polymers. The term "monomer" as used herein refers to a compound that is not polymerized. "polymers" or "biopolymers" may be naturally occurring, chemically modified or chemically synthesized herein.

The term "functional group" generally refers to a compound that may be suitable for chemically bonding first and second molecules together. Chemical binding can be considered to be a broad coverage of binding, which has certain covalent properties, with or without polar binding, and can be characterized by ligand-metal binding along with varying degrees of ionic binding. The functional group may refer to a ligand-receptor binding relationship, where covalent binding is a typical association. The functional group may be selected according to the composition of the molecule. The other functional group of the linker may be adapted to covalently bind the first and second molecules together. Covalent bonding broadly refers to covalent bonding to sigma bonds, pi bonds, hydrogen bonds, other delocalized covalent bonds, and/or other types of covalent bonding, and may be polar bonds with or without ionic bonding components, and the like. The covalent linker comprises a functionalized organic molecule. Functional groups may include hydroxyl groups, amino groups, carboxyl groups, carboxylic anhydride groups, mercapto groups, and silane groups.

The present invention relates to systems and methods for preparing agents, such as therapeutic or imaging agents, embedded or encapsulated in a medium, capsule, particle, mediator, or carrier, in order to maximize treatment and/or imaging of malignant tumors while minimizing adverse effects of the treatment and/or imaging. In particular, the systems and methods of the present invention can form at least one bi-fluid charged jet as depicted in fig. 1, wherein a single capillary tube is used to prepare a capsule with an embedded or encapsulated agent. More particularly, the present invention provides systems and methods for producing capsules and particles having at least one encapsulated and/or embedded agent, such as a therapeutic agent, an imaging agent, and other components.

In one embodiment of the invention, the capsule can serve multiple purposes. First, the capsule protects the encapsulated agent from biological attack and allows it to reach the target site before degradation. Second, the capsule prevents surface interactions of the agent from inducing unwanted signaling pathways and induces the production of a cascade of target intracellular proteins or hormones that limit the efficacy of the agent.

Capsules with embedded or encapsulated agents may be used for the treatment or imaging of cancer in a patient. For example, the capsule can have an embedded or encapsulated therapeutic agent to treat malignant cancers such as lymphoma, leukemia, sarcoma, mesothelioma, glioma, germ cell tumor, and choriocarcinoma. In addition, the capsule may have an embedded or encapsulated imaging agent for detection of cancer. The capsules of the invention may also have a combination of embedded or encapsulated therapeutic and imaging agents.

Fig. 2 is a schematic diagram depicting an embodiment of the present invention showing the general structure of a capsule 200, the capsule 200 having a discrete core region 202 and shell region 204. The core 202 may contain one or more pharmaceutical agents, such as therapeutic agents of biological or non-biological origin. The shell may be made of one or more biocompatible polymers and may contain embedded regions 206 of one or more materials that function to open the shell when the vesicles are in substantially close proximity to the target site or within the target site (e.g., malignant cells). Additionally, the capsule 200 may also serve as a contrast agent for imaging purposes. Non-biological or functional groups 208 may be chemically or physically grafted to the surface of the shell 204, which may facilitate transport to and/or into the target site, e.g., malignant cells, via receptor or surface charge-mediated endocytosis. In particular, for example, malignant cells may overexpress certain receptors, and surface modification of the capsule 200 with functional groups 208 that selectively bind the overexpressed receptors at the surface of the malignant cells may improve the transport of the capsule 200 to and/or into the malignant cells.

It is not necessary that the capsule 200 be spherical, and other shapes and shapes such as rectangular, tubular and ellipsoidal are also suitable for use with the present invention. Additionally, the capsules 200 may be assembled as individual particles, or may be assembled as a series of capsules attached to each other in a chain-like fashion. In particular, if the capsules 200 are assembled in a chain-like series, the capsules may have a length of about 0.03 μm to about 30 μm. Furthermore, it is also effective in case the capsule 200 has a rectangular, tubular or ellipsoidal shape, if the length, e.g. the minor axis in case of an ellipsoid or the diameter in case of a tube, is small enough to pass the target moiety, e.g. a malignant cell membrane. The capsule may have a diameter or minor axis of about 10nm to about 1 μm.

According to one embodiment of the invention, for example, the structure of the capsule may be based on the mass ratio of shell material to core material, as well as the composition and mass of the agent, e.g., therapeutic agent, imaging agent, and other components. In addition, the isolated core and shell regions may not be required. One or more pharmaceutical agents, such as therapeutic or imaging agents, may be embedded and/or dispersed within particles having at least one biocompatible substance, such as a biopolymer. The capsule may include other additives, for example, a chemically functionalized surface that utilizes the biochemical properties of the malignant cell receptor, dispersed particles that help to rupture or separate the capsule carrying the therapeutic and/or imaging agent once the capsule is substantially in close proximity to or within the malignant cell.

According to embodiments of the invention, the capsule may have a shell thickness of about 10nm to about 10 μm, and more particularly about 20nm to about 150 nm. In particular, the capsule shell has a thickness of about 20 nm. Further, the capsule can have a size of about 25nm to about 25 μm, and more particularly about 50nm to about 500 nm. In particular, the capsules may have a size of about 250 nm.

In another embodiment, a capsule comprising at least one agent, such as a therapeutic or imaging agent, can be transported into the target cell by a process such as endocytosis, transformation, or transduction. Once the capsule enters the target cell, the action of the therapeutic agent for treatment can be induced by chemical stress or external radiation. Once activated, the therapeutic agent can induce the desired effect, such as apoptosis of the target cell. The treatment may be genetic material such as nucleotides, RNA, DNA, bacteria, viruses, proteins, nucleotide fragments, nucleotide-encoded gene products, and non-biological agents. Examples of imaging agents may include magnetic particles, photosensitive materials, radioisotopes, and contrast agents.

The capsules of the present invention may have low toxicity magnetic nanoparticles dispersed therein for external magnetic localization of the capsules. In particular, the magnetic nanoparticles may include ferromagnetic materials such as metallic iron and certain metal oxides such as transition metal oxides, as well as rare earth-based magnetic materials. In particular, the magnetic nanoparticles may be magnetite, e.g. Fe, having a size of about 1nm to about 300nm, and in particular a size of about 10nm₃O₄In addition, metallic silver or gold may be used in combination with radioactive emitters such as β, γ, or α emitters for imaging.

With respect to fig. 3, which illustrates an embodiment of the present invention, an upflow EHD system 300 is shown to form at least one bi-fluid charged jet by using a single capillary tube (fig. 1). Here, system 300 may include a syringe pump 302 containing a core fluid, a syringe pump 304 containing a shell fluid, a fluid tank 306 containing a shell fluid 308, a tube 310, a collection tray 312, a power source 314, an electrode 316, a core fluid supply tube 318, a shell fluid supply tube 320, an open end 322 of tube 310, a fluid flow of shell fluid 324, a cone-jet zone 326, and a plume 328. As shown in fig. 3, a syringe pump 302 containing a wick fluid supplies the wick fluid through a supply tube 318 into a tube 310, which tube 310 delivers the wick fluid from an open end 322 thereof, while the shell fluid 308 is delivered by wetting and capillary forces from a fluid slot 306 surrounding the tube 310. The fluid flow of the shell fluid is designated by the numeral 324. The electrode 316 may be placed in a number of locations, such as on the tube 310, in the fluid slot 306, the core fluid supply tube 318, or the shell fluid supply tube 320. The potential of the core fluid and the shell fluid may be the same.

As described below, the throughput of the process can be increased by making a tube that includes more than one function such as tube 310. Depending on the physical and chemical properties of the shell fluid 308 and the core fluid delivered through the tube 310, the height difference between the open end 322 of the tube 310 and the surface of the shell fluid 308 in the fluid slot 306, the ratio of the core fluid and the shell fluid in the upward traveling jet of charged fluid may be changed.

Moreover, in further embodiments, the current collector 312 may be placed over the surface of the shell fluid 308 in the fluid slot 306 in a variety of orientations. For example, if a flat metal surface is used as the current collector 312, the surface of the current collector may be oriented parallel or non-parallel to the surface of the shell fluid 308 in the fluid tank 306, or a different type of current collector structure may be selected, such as a rotating drum.

Plume 328 may contain droplets of the same charge that repel each other and thus travel toward collecting electrode 312 with increasing cross-sectional area. The potential of the core fluid and the shell fluid may be the same because of their intimate electrical contact. The potential and voltage are associated quantities. The potential may depend on the separation between the point where the cone-jet region, such as region 326, is formed and the electrode collection disk 312. The spacing may have a distance in the range of about 1.5 inches to about 15 inches. Further, the voltage used may be in the range of about 0.5kV to about 50 kV.

Turning to fig. 4, which illustrates another embodiment of the present invention, shell fluid 408, tube 310, collection tray 312, shell fluid stream 424, and extractor 414 are shown. The extractor 414, which is an intermediate electrode, may be incorporated to facilitate the formation of a bi-fluid charged jet. The extractor 414 may have an orifice 416 fabricated on the conductor 418 and may be placed a short distance from the open end 422 of the tube 310. The extractor body may be offset from the intermediate potential of those between the tube 310 and the collection pan 312.

In an alternative embodiment, the direction of the charged two fluids ejection need not be upward. Delivery of the wick fluid to produce a downflow of the bi-fluid electrified fluid jet can be achieved similar to capillary force wetting by a tube or conduit. The area after the extractor may subject the charged fluid or solid structure to a full or partial discharge. This configuration may increase the run time for a fluid or solid structure to reach the collection region from the region where the charged meniscus or meniscus is formed, since the smaller charge reduces mobility. The fully or partially discharged fluid or solid structure may be transported to a collection area by using a gas stream.

The concept of wetting the outer wall of a tube or catheter used to deliver the core fluid of a dual-fluid electrified fluid jet may be applied to other configurations. With respect to fig. 5, which illustrates an embodiment of the present invention, there is shown a tube 510 for delivering a core fluid, a second tube 532 for delivering a shell fluid, and a shell fluid flow 524. In addition to the flow rate, the electric field strength and other variables, the spacing and angle 536 between the outer wall of tube 510 and the open end of tube 532 may be adjusted to ensure that the outer wall of tube 510 is sufficiently wetted to form the desired bi-fluid charged meniscus and charged bi-fluid jet. The addition of two or more tubes or conduits for tube 532 to deliver shell fluid may be considered a natural extension of the system shown in fig. 5. The flow rates of the core and shell fluids may be enhanced by the action of gravity, i.e., by placing their respective supply reservoirs at a high distance in terms of the region where a charged two-fluid meniscus forms, which meniscus is formed by the action of a mechanical or digital pump, or driven by a magnetic field if one or both fluids are magnetic. Once the angle 536 and the number of tubes or conduits serving as the purpose of the tube 532 are taken to ordinary degrees of freedom to produce a bi-fluid electrified fluid jet, many other natural extensions of fig. 5 may be achieved.

For example, fig. 6 (which illustrates another embodiment of the present invention) shows a natural extension of this principle in fig. 5. In fig. 6A, 8 tubes or conduits 632 may be used to deliver shell fluid. Tube 632 may be oriented at an angle of about 90 degrees, where the angle is defined in the same manner as defined for angle 536 in fig. 5. Tube 610 delivers a core fluid, which is supplied through core fluid inlet 644, and forms a charged bi-fluid meniscus as both the shell fluid and core fluid are accelerated by the electric field gradient toward collection region 626 or collector electrode, as described above. In fig. 6A, a shell fluid inlet port 640 may be used to first feed the shell fluid into a sleeve 642 (which encases the tube 610) and then through the tube 632. The number of tubes 632 used may depend on factors such as, but not limited to: the physical and chemical properties of the shell and core fluids, as well as process variables such as, but not limited to, the flow rates of the core and shell fluids and the electric field strength and spatial distribution thereof. As shown in fig. 6B, the distance between the open end of the tube 610 and the open end of the tube 632, the cross-sectional area of the tube 610 and the tube 632 may also be determined based on process variables and the physical and chemical properties of the fluids involved. The tubes 632 need not be in contact with each other.

In another embodiment, the concept of wetting the tube or catheter that delivers the core fluid can be extended to other implemented configurations. Fig. 7A, which illustrates an embodiment of the invention, shows a configuration in which a number of tubes 710 are available to deliver wick fluid from their open ends. The tube 710 may be sandwiched between two walls 714. The shell fluid 716 may then be delivered through the space of the adjacent tube 710 (fig. 7B provides the cross-sectional view of fig. 7A). By the action of the applied electric field, charged, bi-fluid menisci and jets can be generated.

Further, intervals 718 and 720 may be adjusted or designed according to process variables such as, but not limited to: two are related to the flow velocity of the fluid and the electric field strength and its spatial distribution. In particular, a more closely charged meniscus is filled, i.e., the smaller the resulting gap 720, the higher the electric field required to form the meniscus. Unless the gaseous environment surrounding the charged meniscus is not air, corona discharge can occur and these can compromise process continuity. Accordingly, one skilled in the art will be able to determine the interval 720 based on these principles, knowledge of the physical and chemical properties of the core and shell fluids, and process variables. For example, if the physical and chemical properties of the core and shell fluids and the values of the process variables require that the spacing 720 be long enough to produce a drip of shell fluid between adjacent tubes 710, fluid deflectors or baffles may be incorporated between adjacent tubes 710 to transport external fluid to the tubes 710. The separation 718 may be determined by a wetting phenomenon that is found between the outer wall of the tube 710 and the shell fluid 716. In addition, the extractor electrode orifice may be placed a short distance from the corresponding bi-fluid charged meniscus, as described above, and may be used to facilitate formation of the bi-fluid charged structure.

With respect to fig. 8A, which illustrates an embodiment of the present invention, it is shown that the arrangement of tubes 810 delivering core fluid may be different from the arrangement shown in fig. 7A-7B. As shown in fig. 8A-8B, the tubes 810 may be sandwiched between a cylindrical sleeve 812 and a rod 814, and a shell fluid 816 may be delivered through the space between adjacent tubes 810. The number of tubes delivering core fluid in fig. 7A-7B and 8A-8B may be determined according to the desired operational throughput. Likewise, the inner diameter of the tube 810 delivering the core fluid may be in the range of about 0.05mm to about 2.0 mm. The diameter of the tube 810 may also be based on the properties of the fluid of interest and the process variables selected such as, but not limited to, the electric field strength and its spatial distribution and flow rate. The cross-section of the tube 810 delivering the core fluid need not be circular because other shapes such as, but not limited to, ellipsoids and polygons can produce charged bi-fluid menisci and jets, and the opening, cross-section of the tube need not be oriented perpendicular to the axis of the tube. The open end of the tube may take other forms such as, but not limited to, conical or angled.

It may be desirable to pack multiple fixtures as shown at 7A-7B side by side to increase product throughput to obtain the benefits of the general principle of wetting the outer walls of the conduits or tubes that deliver the core fluid with the shell-forming fluid. Also, the principle of operation of the device shown in FIGS. 8A-8B may be used to sandwich more than one annular layer of core fluid-delivering tubing or conduit between cylindrical jackets. Fig. 7A-7B and 8A-8B are thus not limiting, as the surfaces of the tubes that can be used to sandwich multiple delivery core fluids need not be cylindrical or flat, as other surfaces having regular or irregular shapes can be used by those skilled in the art.

In one embodiment of the present invention, the concept of wetting the outer wall of a tube or conduit delivering a fluid by the action of forced flow and electric field can be extended to produce multi-fluid charged menisci and jets. With respect to fig. 9, which illustrates an embodiment of the present invention, an arrangement is shown using a fluid containing two coaxial tubes 910 and 916 for delivering a core fluid 914 and a shell fluid 912, and a third tube or catheter 916 for delivering a third fluid. By wetting the outer wall of the outer tube or catheter of the coaxial arrangement of the two-tubes, there is a natural extension of the principle of generating a sheath flow of the third fluid. By way of example, two or more tubes having a cylindrical cross-section may be arranged in a coaxial manner to deliver two or more fluids by forced flow and electric field. Thus, if N is the number of such coaxial cylindrical tubes, (N +1) fluids can be delivered by wetting the outer wall of the outer most cylindrical tube of the coaxial arrangement of N tubes and by the action of forced flow and electric field to form a charged (N +1) -fluid meniscus and jet or jet.

Fig. 10, which illustrates yet another embodiment of the present invention, depicts wetting of the outer wall of a tube or catheter carrying a core fluid with a sheath fluid. The tube 1002 carries the core fluid. The tube 1004 feeds the shell fluid into the porous body 1006. The porous body 1006 delivers the shell fluid by wetting the outer wall of the tube 1002. The volume of the porous body 1006 may wrap around the tube 1002 and contact the tube 1002 at a portion of its outer wall 1008. The selection of the porous material 1006 may be dictated by the physical and chemical properties of the shell fluid. As an example, latex sponges may constitute suitable materials for handling shell fluids, which contain water and other dissolved or suspended components. The type of porous body 1006 and the pore size distribution of the porous body 1006 may be based on the desired flow rates of the core fluid and the shell fluid, as well as the physical and chemical properties of the shell fluid. Examples of the porous material may include silicone-based polymers, polyamide-based crosslinked polymers, melamine-formaldehyde-based polymers, liquid paraffin, polyvinyl alcohol polymers, and poly (1-lactide) acid.

Further, a device comprising an arrangement of multiple tubes or conduits 1002 may be employed. The cross-section of the tube 1002 delivering the core fluid need not be circular, as other shapes such as, but not limited to, ellipsoids and polygons can create charged bi-fluid menisci and jets, and the opening, cross-section of the tube need not be oriented perpendicular to the axis of the tube. The open end of the tube may take other forms such as, but not limited to, conical or angled. The tube 1002 is replaced by a multi-tube arrangement consisting of more than one tube having a cylindrical cross-section arranged in a coaxial fashion, as described above, to deliver more than two fluids by forced flow and electric field. The concept of placing the extractor electrode orifices a short distance from the corresponding charged fluid meniscus can be used, as described above, to extend the system described in fig. 10 and facilitate the formation of multi-fluid charged structures.

Turning to fig. 11, which illustrates another embodiment of the present invention, the delivery of the shell fluid by the wetting phenomenon is described. The reservoir 1102 for the core fluid may be rotated by any mechanical means (not shown) while partially immersed in the bath 1104 of shell fluid. The shell fluid may wet the outer wall 1108 of the reservoir with the wick fluid by the action of capillary forces, among other physical phenomena. In the region where wetting of the outer wall of the reservoir 1102 by the shell fluid occurs, at least one of an orifice 1106, a capillary or other conduit shape protruding from the outer wall of the reservoir 1102 may be fabricated in the outer wall 1108 of the reservoir 1102 to deliver the core fluid through the shell fluid film, when an electric field is applied between the reservoir 1102 and the collection region or collector electrode. The distance between adjacent orifices 1106, the capillary or conduit shape, and the rotational speed of the reservoir 1102, the type, size and shape of the reservoir 1102 material, the depth of immersion of the reservoir 1102 into the bath 1104, the number and size of the orifices 1106, the capillary or conduit shape may be based on the physical and chemical properties of the core and shell fluids and process variables such as, but not limited to, the voltage used between the reservoir 1102 and the collection region or collector electrode.

In one embodiment, once the dual-fluid electrified fluid jet is formed by the method of the present invention, the plurality of fluids may be processed into different shapes such as, but not limited to, hollow fibers and capsules. A family of chemical synthesis methods known as sol-gel may be used to produce this structure from a bi-fluid charged jet. For example, one way to prepare a structure with defined core and shell regions of different composition requires that the shell fluid undergo full or partial solvent evaporation during the time from the ejection of the bi-fluid charged jet from the bi-fluid charged meniscus to the collector electrode or collection region. The precursor of the solid shell may remain in solution until a critical portion of the solvent evaporates into the environment. The core fluid, depending on how it is formulated by the skilled artisan, may or may not produce a solid phase upon collection at the collection area.

The method of the present invention can be used to produce structures having defined core and shell regions from a wide variety of fluids having different chemical compositions. Additional functions such as, but not limited to, magnetic properties may be added to the core fluid or shell fluid, or both, by chemical action that forms the fluid.

For example, the shell of a capsule containing a core fluid with dissolved proteins can be made magnetic by suspending magnetic particles in the range of about 100nm to about 1nm into a shell fluid precursor prior to processing using the methods described herein. The core fluid and shell fluid chemistry may be prepared in such a way that neither the core fluid nor the shell fluid undergoes solidification prior to entering the droplet-in-droplet mode shown in fig. 3. Once the droplet-in-droplet method is used, evaporation of a critical amount of solvent in the shell fluid is performed and the capsule with solid shell and fluid core is collected in the collecting electrode.

A wide variety of proteins, DNA materials, cells and other substances of biological origin can be encapsulated using the methods of the invention. In particular, enzymes are a class of proteins that can be used as biocatalysts and can be encapsulated in a fluid core such as, but not limited to, a pH buffered aqueous solution, and the shell can be formed from a sol-gel precursor with suspended magnetic particles. Once suspended in the fluid medium, the magnetite portion in the shell may allow magnetic transfer of the capsule. The capsule containing the encapsulated enzyme may be suspended in a fluid containing reactants and products for reaction catalyzed by the encapsulated enzyme. If the shell is designed with pores large enough to allow the diffusion of reactants and products through the pores for the reaction catalyzed by the encapsulated enzyme, but not large enough to allow the enzyme to diffuse out of the capsule, the encapsulated enzyme may be recovered with the aid of magnetic force after the reaction reaches the desired degree.

For example, in one embodiment of the invention, when controlled, the silicon alkoxide sol-gel chemistry may create pores in the capsule of about 0.5nm to about 2.0nm, which may be sufficient to allow diffusion of many commercially important reactants and products, but will not allow, for example, about 50kDa to about 60kDa transaminase to escape the sol-gel derived shell. A natural extension of this idea is to design a core fluid formulation containing biologically derived material other than proteins such as, but not limited to, DNA fragments, genes, and cells.

According to one embodiment, it is possible to produce magnetite nano-particle formulations compatible with silicon alkoxide-based shell chemistry, which may be encapsulated with a silicon alkoxide layer having about 2nm to about 4 nm. This silanol layer makes it suitable for strongly anchoring the magnetite to the shell solid phase by a cross-linked sol-gel operation. The specific mass part of the magnetite phase in the shell ranges from about 0.1 to about 0.8. Fig. 12 is a photograph of a charged fluid meniscus (with added magnetite particles) formed under the electric field and forced flow created by the shell.

In another embodiment of the present invention, capsules suitable for the treatment and/or imaging of malignant cancers may be prepared by using electrospray. In general, the electrospray process may comprise delivering a fluid containing an encapsulate through a single nozzle, capillary, conduit or orifice, applying an electrical potential between the nozzle and a collection region (hereinafter "counter-electrode"), forming a DC electrospray for the fluid containing the encapsulate, accelerating the DC electrospray towards the counter-electrode, forming a shell encapsulating the DC electrospray droplet to produce a small bubble. The vesicles may be formed during the flight of the DC electrospray from its delivery region to the counter-electrode by reaction between suitable shell-forming monomers and fluid components in the electrospray process. The shell-forming monomers and fluid components do not compromise the integrity and function of the encapsulate. The capsules can be collected on counter-electrodes and used for storage for further use.

With respect to fig. 13A, which illustrates an embodiment of the present invention, a schematic of an electrospray process of the present invention is provided. The electrospray method is based on a combination of chemical reactions between the electrospray and its surrounding gas phase. In fig. 13A, an electrospray nozzle 1302 and a cloud 1304 are depicted. In addition, an inlet 1306 and an outlet 1308 are shown, a counter-electrode 1310, a shell-forming reaction zone 1312, and a high-voltage DC source 1314. Inlet 1306 may fill the shell-forming region or cavity with a gas containing shell-forming monomers. The electrospray process of the present invention provides a simple and scalable single-fluid electrospray process, thereby completely avoiding the use of multiple coaxial capillary nozzles. Since electrospray can be produced at low reynolds numbers, i.e. under low shear conditions, the electrospray process of the present invention produces true core-shell capsules without the low yield and high shear process loss of the emulsion-polymerization process. In addition, the method also allows for the production of smaller capsules than those produced by AC single-fluid electrospray.

In one embodiment, the electrospray process may be spatially delivered to a region where the gaseous environment has a suitably concentrated shell-forming monomer. As shown in fig. 13A, the electrospray process may be continuous or semi-continuous, depending on whether the supply of the electrospray encapsulate containing fluid, the supply of the monomer containing gas and the removal of the encapsulate material are all continuously prepared. Since the encapsulated fluid core cannot deliver the component reacting with the reactive monomer by diffusion to the outside, forming the shell indefinitely, the shell formation process should be terminated after a certain shell thickness is reached, so the shell thickness of the vesicles can be controlled by suppressing many process variables such as, but not limited to: monomer concentration, concentration of reactive species in electrospray, temperature, time of flight of electrospray, voltage, electrospray mean droplet size, and monomer and electrospray chemistry.

In another embodiment of the invention, electrospray containing encapsulates may be through a solution containing shell-forming monomers, rather than a gas containing the monomers.

In another embodiment, the electrospray process of the present invention may be discontinuous and the time of ejection of the charged suspension in the monomer-containing gas space may be extended by the use of a high voltage AC potential within the shell-forming reaction volume, as shown in fig. 13B. With respect to fig. 13B, an AC electrode 1316 and an AC power supply 1318 are shown. As shown in fig. 13A, the fluid or solution containing the encapsulates is first made into an electrospray cloud, but during certain time points of the electrospray operation, the DC high voltage and the fluid flow of the encapsulates containing the fluid or solution are switched off while the AC voltage is turned on. The charged droplets may then be made to oscillate between the two AC electrodes 1316 and the frequency of the AC field is set so as to prevent the charged droplets from striking one AC electrode 1316. After the desired amount of time has elapsed, the AC high power supply 1318 is turned off while the DC voltage is turned back on, thereby collecting the bubbles in the DC counter-electrode 1310. By extending the time, the electrospray remains suspended in the area of the shell forming the vesicles, the reaction can be controlled, and a greater range of shell formation times can be achieved than with the device shown in fig. 13A. The natural expansion of the vesicle generation protocol shown in fig. 13B can replace the gaseous environment in the shell-forming cavity with an insulating fluid or solution bath containing the shell-forming monomer or monomers.

In an alternative embodiment of the invention, a single fluid charged jet may be used in the form of a solution or emulsion to produce a capsule with at least one embedded pharmaceutical agent. As used herein, an emulsion generally refers to a single fluid that can be used in the EHD process described above to produce the particles rather than capsules, wherein the same concentration range of the therapeutic and other components of the capsules, as described below, and the same average particle size are used. In addition, magnetic nanoparticles may also be dispersed in the particles.

For example, an aqueous emulsified phase containing at least one pharmaceutical agent, such as a therapeutic agent and/or an imaging agent, and stabilizing salts and polyelectrolytes may be dispersed in an organic solution containing at least one biopolymer. The resulting emulsion comprises a biphasic medium which does not form a separate layer in the formation of the charged jet. The aqueous phase may be formulated in the same manner as the core fluid in the case of the capsules described above, and the biopolymer or biopolymer containing organic phase may also include magnetic nanoparticles and/or optoelectronic particles formulated as described above for the method of formulating the shell fluid of the capsules. Alternatively, using certain co-solvents such as, but not limited to, dimethyl sulfoxide, a single fluid charged jet formulation results in the generation of a homogeneous fluid mixture or solution comprising the biopolymer matrix precursor or precursors and the active component and receptor. A single charged fluid jet process can result in an embedded, rather than encapsulated, active agent.

According to one embodiment, for example, the shell-forming monomer may comprise a compound in the cyanoacrylate family, such as an alkyl-substituted cyanoacrylate. Furthermore, during the flight time of the electrospray to the counter-electrode, the formulation containing the encapsulate may be a compound that readily reacts with the cyanoacrylate monomer to give a skin of the shell without chemically altering the intended function of the encapsulate. In particular, the encapsulate may contain water. The encapsulate may be of biological origin, such as nucleotides, DNA fragments thereof, RNA fragments thereof, proteins, lipids, carbohydrates, combinations thereof and/or any modifications thereof. Additionally, the encapsulate may include a non-biological therapeutic drug as described below.

According to one embodiment of the present invention, suitable biopolymers for producing the shell of EHD-derived capsules include, without limitation, PLA, PLGA, chitosan, alkyl methacrylates, and other bioabsorbable polymers such as PCL, starch, and copolymers derived from any combination thereof. In addition, the PEG may also be incorporated into one or more other biopolymers as a polymer chain or copolymer segment, or functionalized with groups such as: thio, carboxylic acid and amine groups. In addition, other compositions may be incorporated into the shell fluid prior to electrospray, such as nano-sized magnetite particles having a size range of about 1nm to about 300nm, and surfactants to produce magnetically sensitive MRI materials. Further, nanosized photo-sensitive silver, gold, palladium, or combinations thereof having a size range of about 1nm to about 50nm may be added to the shell precursor fluid prior to electrospray to produce a photo-sensitive material.

According to another embodiment, a core fluid suitable for use in the methods of the present invention may comprise an aqueous phase having a buffer salt and a polyelectrolyte such as phosphate buffer to structurally stabilize a solution of biologically derived therapeutic agents, one or more therapeutic agents such as, but not limited to, tumor necrosis factor α (TNF- α) protein or TNF- α, a cDNA encoding for TNF- α or Egr-TNF, one or more substances capable of inducing the action of Egr-TNF such as, but not limited to, temozolomide or TMZ, and a chemotherapeutic agent such as, but not limited to, acridine anisidine (acridinyl aniidide), allopurinol, altretamine, aminoglutethimide, androgen, diarsenium trioxide, asparaginase (asparaginase), azacitidine, angiogenesis inhibitors, bleomycin, bortezomib, busulfan, capecitabine, carboplatin, carmustine, cetuximab, cisplatin, chlorambucilazalomustine, cladribine, dactinomycin, valtretinomycin, valtrexadine, doxorubicin, valtrexadine, valtrexatin, doxorubicin, doxorubicine.

In addition, the capsules or particles of the present invention may contain chemotherapeutic enhancing agents such as, but not limited to, kexijun, and chemotherapeutic protective agents such as, but not limited to, amifostine certain radioactive elements may be useful in the treatment and/or imaging of cancer and in one embodiment of the present invention radioactive elements may be incorporated into the electrospray-produced particles.for example, the actinium-227 isotope precursor of radium-223 is the so-called α for the treatment of metastases in bone from breast and prostate cancer and for radioimmunotherapy, copper-67, a gamma and β emitter for cancer radioimmunotherapy and lymphoma and colon cancer and breast cancer diagnosis, iodine-131 for the treatment of breast, leukemia and lymphoma using radioimmunotherapy, thyroid function and disease (e.g., cancer) studies and benign thyroid diseases (e.g., hyperthyroidism), iodine-125 for use as an implant for breast and prostate tumors, radiolabels, radiation therapy, osteoporosis detection of bone, tracer drugs, diagnostic agents, brain receptor imaging, rhenium and prostate cancer, for the diagnosis of prostate cancer, renal adenocarcinoma, and renal adenocarcinoma for the radioimmunotherapy, and renal adenocarcinoma for the sequential radioimmunotherapy of renal cancer, and renal-lymphoma, and renal-90 for the detection of renal-renal cell cancer, and renal cell transplantation.

In further embodiments, radioisotopes may be incorporated into the capsules of the invention and their most common use in medicine and related fields is cadmium-109 for general cancer detection, actinium-225 and thorium-229, the latter being precursors of actinium-225 and progenitors of bismuth-213, which are α emitters for cancer radioimmunotherapy, bismuth-212 and thorium-228, the latter being bismuth-212 precursors, which are α emitters for cancer radioimmunotherapy, cobalt-60, which is the radioactive source of cancer radiotherapy, for irradiation of food and medical material supplies, copper-64, a cationic emitter for cancer therapy, dysprosium-166 for cancer radioimmunotherapy, erbium-169 for small joint rheumatoid arthritis therapy, europium-152 and europium-154, which is the radioactive source of medical material supplies and food irradiation, gadolinium-153 for evaluation of osteoporosis, gold-198 for prostate and brain cancers, and for cancer therapy and for bone and brain cancer therapy and for bone marrow transplantation, as well as a radiographic marking agent for bone marrow-188, calcium-150 for bone marrow transplantation, calcium-188, calcium-phosphate-188, calcium-phosphate-188, calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium-phosphate-calcium.

In another embodiment, the capsule of the present invention may comprise at least one surface biochemical that may be chemically or physically attached to enhance the affinity of the capsule for glioma cells, which may comprise epidermal growth factor or EGF. Epidermal growth factor may be incorporated into the surface of the capsule as part of the shell fluid precursor after or during the EHD treatment. In particular, the content of EGF in the capsules and particles prepared by the method of the present invention may range from about 0 to about 0.02 wt%. In addition, other surface biochemicals and chemicals can include interleukin-13, interleukin-4, peripheral benzodiazepines, vascular endothelial growth factor, platelet-derived growth factor, fibroblast growth factor, urokinase-type plasminogen activator, folic acid and folic acid derivatives, neurotrophic factor growth factor, somatostatin, cBSA, cHSA, protamine, insulin, ApoE-derived peptides, polysorbate-80, OX-26, delivery agents (transferring), glucose, mannose, RMP-7, thiamine, combinations thereof, and functional equivalents thereof. The surface biochemicals can be of human, animal, or recombinant origin, and for non-estrogen based surface biochemicals have a content range of about 0 to about 1 wt%.

In further embodiments, the capsule may include at least one surface biochemical that may be chemically or physically attached to enhance the affinity of the capsule for lymphomas, myelomas, and leukemias, such as VEGR-2, tositumomab, a monoclonal antibody that binds the CD20 receptor, and other antibodies and monoclonal antibodies particularly those designated as other differentiation antigen clusters, such as, but not limited to, CD5, CD7, CD13, CD19, CD22, CD33, CD52, and CD61, as well as anti-myeloperoxidase, any combination thereof, and functional equivalents thereof. These biochemicals and their derivatives can be incorporated into the surface of the capsule as part of the shell fluid precursor after or during the EHD process. The amount of surface receptors in the capsule ranges from about 0.0 to about 1.0 wt%. Other biochemicals and chemicals that can be incorporated into the capsule can include tyrosine kinase receptors, including fibroblast growth factor receptor (FGFR-1), EGF, TGF, VEGF-A, urokinase receptor, interleukin 4 receptor, retinoic acid receptor, heparin-binding EGF-like growth factor or HB-EGF, amphiregulin, epidermal regulator (epiregulin), neuregulin, human epidermal growth factor 2(HER-2) and family members, erbB2 and erbB1 receptor families, interleukin-13 and family derivatives, platelet-derived growth factor, urokinase-type plasminogen (plasminon) activators, folic acid and folic acid derivatives, neurotrophic factor growth factor, somatostatin, combinations thereof, and functional equivalents thereof. The surface biochemicals can be of human, animal, or recombinant origin and are present in an amount ranging from about 0.0 to about 1.0 wt%.

In yet a further embodiment, the capsule may include at least one surface biochemical substance that may be chemically or physically attached to enhance the affinity of the capsule for breast cancer, such as estrogen. Estrogens and chemical derivatives thereof may be incorporated into the surface of the capsule as part of the shell fluid precursor after or during the EHD process. The amount of estrogen or derivative thereof in the capsules and particles ranges from about 0.02 to about 0.4 wt%. Other surface biochemicals and chemicals that can be incorporated into the surface of the capsules and particles can include human epidermal growth factor 2(HER-2) and family members, erbB2 and erbB1 receptor families, interleukin-13 and family derivatives, vascular endothelial growth factor, platelet-derived growth factor, fibroblast growth factor, urokinase-type plasminogen activator, folic acid and folic acid derivatives, neurotrophic factor growth factor, somatostatin, combinations thereof, and functional equivalents thereof. The surface biochemicals can be of human, animal, or recombinant origin and can be present in an amount ranging from about 0.0 to about 1 wt%.

In another embodiment, the capsule may include a surface biochemical that may be chemically or physically attached to enhance the affinity of the capsule for pancreatic cancer cells, may be a tyrosine kinase receptor, including fibroblast growth factor receptor (FGFR-1), EGF, TGF, VEGF-A, urokinase receptor, interleukin 4 receptor, retinoic acid receptor, heparin-binding EGF-like growth factor, or HB-EGF, amphiregulin, epiregulin, neuregulin, and functional equivalents thereof. These biochemicals and their derivatives can be incorporated into the surface of the capsule as part of the shell fluid precursor after or during the EHD process. The content of the surface receptors in the capsules and particles ranges from about 0.0 to about 1.0 wt%. Other surface biochemicals and chemicals that can be incorporated into the surface of the capsules and particles can include human epidermal growth factor 2(HER-2) and family members, erbB2 and erbB1 receptor families, interleukin-13 and family derivatives, platelet derived growth factor, urokinase-type plasminogen activator, folic acid and folic acid derivatives, neurotrophic factor growth factor, somatostatin, combinations thereof, and functional equivalents thereof. The surface biochemicals can be of human, animal, or recombinant origin and can be present in an amount ranging from about 0.0 to about 1 wt%.

In addition, other surface biochemicals and chemicals for the capsule and particle can be fused into signal sequences, mitochondria, nucleus, actin and tubulin, golgi, plasma membrane, peroxisomes, and can be in the content range of about 0.0 to about 1.0 wt%.

For example, according to one embodiment of the invention, the concentration of paclitaxel in the capsule or particle may be in the range of about 10 to about 3200 μ g/mL, particularly, the concentration of TNF- α protein in the capsule or particle may be in the range of about 100 μ g/mL to about 1,000 μ g/mL, more particularly, the concentration of Egr-TNF in the capsule or particle may be in the range of about 10 μ g/mL to about 300 μ g/mL, the therapeutic effect may also be achieved with capsules containing one or two compounds of the three-component group of paclitaxel, TNF-a, and Egr-TNF, the narrow range of mass ratios of therapeutic and other capsule components, the average capsule size, the average shell thickness, and the narrow core to shell average mass ratio may be selected based on the location and size of the malignant tumor, and the use of doped magnetic nanoparticles to help direct the capsules to the malignant tumor may not be needed, such as direct tumor injection, may be used in place of a magnetic helper device, and the irradiation of the capsule with the present invention may be used to cause the luciferase to be released by electromagnetic radiation of the MCT-DNA of the capsule or of the luciferase-DNA that may be used as a luciferase-induced by electromagnetic luciferase-induced by the electromagnetic luciferase-induced luciferase gene inserted into the electromagnetic luciferase-DNA-plasmid-DNA-vector used in the process, such as, but not limited to induce the luciferase gene-expressing the luciferase gene that is used in the luciferase-.

In alternative embodiments, enhanced convection delivery or CED may be used in place of magnetic assisted delivery, and contact of harmful electromagnetic radiation with the particles or capsules prepared by the methods of the invention may be used to trigger lysis of the capsules or particles with release of therapeutic agents to trigger biochemical processes such as, but not limited to, malignant cellular DNA damage, or a combination of processes induced by the electromagnetic radiation.

Agents suitable for encapsulation or embedding in the capsules of the present invention should not be construed as limited to agents suitable for cancer therapy or malignant tumor imaging, as described above. However, alternative agents that may be suitable for encapsulation or encapsulation may include anti-inflammatory compounds, anti-allergic agents, glucocorticoids, anti-infective agents, antibiotics, antifungal agents, antiviral agents, mucolytics, antiseptics, vasoconstrictors, wound healing agents, local anesthetics, peptides, and proteins.

Examples of potentially useful anti-inflammatory compounds are glucocorticoids and non-steroidal anti-inflammatory agents such as betamethasone, beclomethasone, budesonide, ciclesonide, dexamethasone, desoximetasone, fluocinonide (fluocinolone acetonide), fluocinonide (flucinonide), flunisolide, fluticasone, echocortolone (icomthaseone), rofleponide, triamcinolone acetonide, fluccorbutine, hydrocortisone-17-butyrate, prednisolone, 6-methylprednisolone aceponate, mometasone furoate, elastane-, prostaglandins-, leukotrienes-, bradykinin-antagonists; non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen, indomethacin, including any pharmaceutically acceptable salt, ester, isomer, stereoisomer, diastereoisomer, epimer, solvate or other hydrate, prodrug, derivative or any other chemical or physical form of the active compound comprising the respective active moiety.

Examples of potentially useful antiallergic agents include the aforementioned glucocorticoids, as well as nedocromil, cetirizine, loratadine, montelukast, roflumilast, zileuton, omalizumab, heparin and heparin analogs as well as other antihistamines, azelastine, cetirizine, desloratadine, ebastine, fexofenadine, levocetirizine, loratadine.

Anti-infective agents, the classes or therapeutic classifications thereof being understood herein to include compounds effective against cellular, fungal and viral infections, i.e. including antimicrobials, antibiotics, antifungals, antiseptics and antivirals, the anti-infective agents being penicillins including benzylpenicillins (penicillin-G-sodium, clemizone penicillin, benzathine penicillin G), phenoxypenicillins (penicillin V, propicillin), ampicillins (ampicillin, amoxicillin, bacampicillin), acylaminopenicillins (azlocillin, mezlocillin, piperacillin, apacillin), carboxypeptins (carbenicillin, ticarcillin, temocillin), isoxazolpenicillins (benzizocillin, cloxacillin, dicloxacillin, flucloxacillin), and amidine (amiridin) penicillins (mecillin), cephalosporins including cephalosporins (cefazolin, cefuroxime), cefepime (cefepime ), cephalosporins (cefepime ), cefepime (cefepime ), cefepime (cefepime), cefepime (cefepime), cefepime (cefepime), cefepime (cefepime), cefepime (cefepime), cefepime (cefepime), cefepime (cefepime), cefepimeIsepamicin, arbekacin, tobramycin, netilmicin, spectinomycin, streptomycin, capreomycin, neomycin, paromomycin (paromomycin), and kanamycin; macrolides including erythromycin, clarithromycin (clarithromycin), roxithromycin, azithromycin, dirithromycin (dithromycin), josamycin, spiramycin, and telithromycin; gyrase inhibitors or fluoroquinolones (fleroquinolones) including ciprofloxacin, gatifloxacin, norfloxacin, ofloxacin, levofloxacin, mefloxacin, lomefloxacin, fleroxacin, ganofloxacin, clinafloxacin, sitafloxacin, prulifloxacin, olafloxacin, cadefloxacin, gemifloxacin, balofloxacin, trovafloxacin and moxifloxacin; tetracyclines, including tetracycline, oxytetracycline, rolicycline, minocycline, doxycycline, tigecycline, and aminocyclin; glycopeptides including vancomycin, teicoplanin, rituximab, avoparcin, oritavancin, ramoplanin, and peptide 4; polypeptides including plectasin, dalbavancin, daptomycin, oritavancin, ramoplanin, dalbavancin, telavancin, bacitracin, brevibactein, neomycin, kanamycin, mupirocin, paromomycin, polymyxin B, and colistin; sulfonamides including sulfadiazine, sulfamethoxazole, sulfalene, sulfamethoxazole, co-trimetrol, co-trimoxazine and co-tetraxazine; azoles including clotrimazole, oxiconazole, miconazole, ketoconazole, itraconazole, fluconazole, metronidazole, tinidazole, bifonazole, raviconazole, posaconazole, voriconazole and ornidazole, and other antifungal agents including flucytosine, griseofulvin, tolnaftate, naftifine, terbinafine, amorolfine, ciclopirox olamine, echinocandins such as micafungin, caspofungin, anidulafungin; nitrofurans including nitrofurantoin and nitrofurazone; polyenes including amphotericin B, natamycin, nystatin, flucytosine; other antibiotics include tithromycin, lincomycin, clindamycin, and the like,Oxazolidinones (linzezolids), rabezolids, and mixtures thereof,Streptogramins eA + B, pristinamycin aA + B, virginiamycin A + B, dalfopristin/quinupristin (syndesmin), chloramphenicol, ethambutol, pyrazinamide, terizidone, dapsone, propylthioisoniamide, fosfomycin, fusidic acid, rifampin, isoniazid, cycloserine, terizidone, ansamycin, lysostaphin, elaprine, mirocin B17, clorocidin, filgrastim and pentamidine, antiviral agents including acyclovir, ganciclovir, bivudin, valacyclovir, zidovudine, dideoxyadenosine, thiacytidin, stavudine, lamivudine, zacitabine, ribavirin, nevirapine (nevirapirimapirimirin), delavirdine (aviridine), allantoin, ritonavir, quinavirus, quinavirenza, quinavirenzamide, picrinine, picroritin, picroritinib, picrinine, picroritin, picrinine, geraniol, geraniin, geraniol.

Examples of mucolytics that may be useful are DNase, P2Y 2-agonists (denufosol), heparin analogues, guaifenesin, acetylcysteine, carbocisteine, ambroxol, bromhexine, lecithin, myrtle oil and recombinant surfactant proteins.

Examples of potentially useful local anesthetics include benzocaine, tetracaine, procaine, lidocaine, and bupivacaine.

Examples of potentially useful antiallergic agents include the aforementioned glucocorticoids, nedocromil. Examples of potentially useful peptides and proteins include antibody agonist toxins produced by microorganisms, antimicrobial peptides such as moth-blood, defensins, thionin, and cathelicidin.

Furthermore, immunomodulators include methotrexate, azathioprine, cyclosporin a, tacrolimus, sirolimus, rapamycin, mycophenolate mofetil (mycophenolate), moroxydine, cytotatics, and metastasis inhibitors, alkylating agents such as nimustine, melphalan (melphanlane), carmustine, lomustine, cyclophosphamide (cyclophosphamide), ifosfamide, trofosfamide, chlorambucil, busulfan (busulfane), treosulfane (treosulfane), punicin, thiotepa; antimetabolites such as cytarabine, fluorouracil, methotrexate, mercaptopurine, thioguanine; alkaloids such as vinblastine, vincristine, vindesine; antibiotics such as alcaubicine, pingyangmycin, actinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mitomycin, plicamycin; group II element (e.g., Ti, Zr, V, Nb, Ta, Mo, W, Pt) complexes such as carboplatin, cisplatin, and metallocene compounds such as titanocendichloride; amsacrine, dacarbazine, estramustine, etoposide, beraprost, hydroxyurea, mitoxantrone (mitoxantrone), procarbazine, teniposide (temiposide); paclitaxel, iressa, zactima, poly-ADP-ribose-polymerase (PRAP) enzyme inhibitors, carnosol anthraquinone, gemcitabine, pemetrexed, bevacizumab, ranibizumab may be suitable for inclusion or encapsulation in the capsules of the invention.

In further embodiments, other compounds may include protease inhibitors such as alpha-anti-trypsin, antioxidants such as tocopherols, glutathione, pituitary hormones, hypothalamic hormones, regulatory peptides and other inhibitors, corticotropin, techoxin, chorionic gonadotropin, urogonadotropin, saomototropipine, ergometrilin, desmopressin, oxytocin, argininopressin, ornipropressin, leuprolide, triptorelin, gonadorelin, buserelin, nafarelin, goserelin, parathyroid hormone, calcium metabolism regulators, dihydrotachysterol, calcitonin, clodronic acid, etidronic acid, thyroid therapeutics, sex hormones and inhibitors thereof, protein assimilating agents, androgens, antiestrogens, progestins, antiestrogens, antimigraine drugs such as propoxypharbide, lisuride, meclizine, ergotamine, mepiquide, ergotamine, clonidine, benzothiophenine (CSF), hypnotic drugs such as, mepiquinone, diethylstilbenine, diethylstilbestrol, diethylcarbamazepine, e, antihistamines, e, pheromones, e, antihistamines, neuroleptinones, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-e, e-.

The description and examples set forth above are intended to be illustrative only and are not intended to be exhaustive of all possible embodiments, applications, or modifications of the invention. Thus, various modifications and variations of the described methods and systems of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. While the invention has been described in conjunction with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in the materials science, polymer science or related fields are intended to be within the scope of the following claims.

The disclosures of all references and publications cited above are expressly incorporated by reference in their entirety to the same extent as if each were incorporated by reference.

Brief Description of Drawings

The accompanying drawings, which are included to provide a further understanding of the invention, are incorporated in and constitute a part of this specification; the illustrative embodiments of the invention together with the detailed description serve to explain the principles of the invention. No attempt is made to show structural details of the invention in more detail than is necessary for a fundamental understanding of the invention and the various ways in which it may be practiced.

FIG. 1 is a schematic diagram showing a bi-fluid charged jet, which is a fluid stream comprising at least two distinct fluid layers, according to the principles of the present invention.

Figure 2 is a schematic diagram showing the general structure of a capsule produced by the method of the invention.

Fig. 3 is a schematic diagram illustrating an upstream electrohydrodynamic system according to the principles of the present invention.

Fig. 4 is an exploded schematic view of the upstream hydrodynamic system of fig. 3, showing the addition of an extractor (extractor).

FIG. 5 is a schematic diagram showing a downwash electrohydrodynamic system using a single capillary tube to form a bi-fluid charged jet in accordance with the principles of the present invention.

Fig. 6A is a schematic diagram depicting a downflow electrohydrodynamic system using a plurality of tubes or conduits to deliver the shell fluid in accordance with the principles of the present invention.

Fig. 6B is a cross-sectional view of fig. 6A.

FIG. 7A is a schematic diagram illustrating a downflow hydrokinetic system using a plurality of capillaries between two plates to deliver the core fluid in accordance with the principles of the present invention.

Fig. 7B is a cross-sectional view of fig. 7A.

FIG. 8A is a schematic diagram showing a downflow electrohydrodynamic system using a plurality of capillaries to deliver the core fluid in accordance with the principles of the present invention.

Fig. 8B is a cross-sectional view of fig. 8A.

FIG. 9 is a schematic diagram showing a descending liquid electrohydrodynamic system for producing a capsule having at least three layers in accordance with the principles of the present invention.

FIG. 10 is a schematic diagram showing a downflow hydrohydrodynamic system using a porous body to deliver the shell fluid to create the dual-flow Taylor cone according to the principles of the present invention.

Fig. 11 is a schematic diagram of an upstream electro-hydrodynamic system in which the core fluid reservoir may be rotated in accordance with the principles of the present invention.

Fig. 12 is a photograph of a charged meniscus formed under the forced flow and electric field of the shell formulation.

Fig. 13A is a schematic diagram depicting the method of the present invention based on the combination of electrospray and chemical reaction between its surrounding gas phase, in accordance with the principles of the present invention.

Fig. 13B is a schematic diagram depicting the method of the present invention based on the combination of electrospray and chemical reaction between its surrounding gas phase, in accordance with the principles of the present invention.

Fig. 14 shows a photograph obtained by confocal microscopy. Panel I is the unfocused signal and panel II shows the focused signal in the capsule core fluid region.

FIG. 15 is a graph showing the activity of the transaminase in encapsulation as well as in free solution when converting optical rotation to equivalent D- (or L-) glutamine concentrations by a standard curve.

Detailed Description

Specific example 1:

this example describes the encapsulation of a protein solution using a modified formulation of a shell formulation consisting of a solution of Bovine Serum Albumin (BSA) in which a salt such as phosphate is present to stabilize the pH of the solution. Confocal Fluorescence Microscopy (CFM) and BSA with fluorescent labels were used to image the capsules. About 2mg of fluorescent protein was dissolved in about 1mL of Phosphate Buffered Saline (PBS), which is a typical buffer to stabilize this aqueous solution to an acid-base pH equal to about 7.4.

The final BSA concentration was adjusted to about 3 μ M. The sol aging procedure was carried out before using silica sol as shell fluid precursor, adding t-amyl alcohol to 50:50 volume ratio.

Typically, for solution aging purposes, the acidified tetraethyl orthosilicate solution in ethanol is aged at about 80 ℃ for about 4 to about 6 hours. T-amyl alcohol was added to increase the hydrophobicity of the shell fluid, which further prevented any significant mixing between the core fluid and the shell fluid.

The core and shell flow rates were adjusted to about 0.025 and about 0.75 ml/hr, respectively. Sufficient voltage for capsule design is in the range of about 11 to about 12kV and the distance between the bi-fluid charged meniscus and the collection zone is in the range of about 4 to about 14 cm. The collection region or collector is a flat metal surface held at ground potential while the two fluid charged menisci are held at a positive electrical bias in the range of about 5 to about 18 kV. Fig. 14 shows a photograph taken with the help of CFM techniques, where the limitation of BSA is evident when the instrument is focused to generate a fluorescent signal in the core fluid region of the capsule. With respect to fig. 14, panel I shows the unfocused fluorescence signal, while panel II shows the focused fluorescence signal.

Specific example 2:

the sol-gel process and process variables used to encapsulate the fluorescent BSA in example 1 were modified slightly to encapsulate the transaminase. This enzyme is used to catalyze the following reactions:

d, L-Glutamine + glyoxylic acid → L-Glutamine (left unreacted) + α -one derivative (from D-Glutamine) + Glycine

Each enantiomeric form has the ability to rotate polarized light and a technique known as polarimetry can be used to follow chemical reactions involving enantiomers as a function of time. The reactant to the left of the reaction shown above is essentially a mixture that is not optically active, since the D, L prefix represents an approximately 50:50 mixture of the D and L enantiomers of glutamine. The product on the right of the reaction shown above becomes enriched in unreacted L-glutamine, accompanied by a time-dependent optical rotation signal which can be traced by optical rotation analysis, since this particular transaminase only catalyzes reactions related to the D enantiomer. The reaction was buffered to a pH of about 7.5 with PBS.

Because of the low optical rotation of the glutamine isomer in pure water, experimental protocols were designed to stop the reaction before the quantitation of the optical rotaline analysis at different reaction times and to increase the sensitivity of the analytical technique. This was achieved by adding about 1.0mL of 37wt% HCl to the aliquot removed from the reactor, which denatures the enzyme. The enzyme catalyst sinks to the bottom of the bottle or is removed by centrifugation; acidification increases the sensitivity of the polarimetric technique by about the order of 1/2 compared to the results observed in nearly neutral solutions.

Biocatalytic testing was performed using a substrate concentration of about 50mM and an enzyme concentration of about 0.5 mg/mL. FIG. 15 shows an example of the activity of this transaminase in the encapsulated state and free in solution, once the optical rotation has been converted to the equivalent D- (or L-) glutamine concentration by a previously determined standard curve.

Specific example 3: particles with 3.1LACZ as DNA marker

A therapeutic agent solution was prepared by mixing 3.1LACZ in about 10mM Bis-Tris propane buffered water containing about 1wt% isopropanol and about 2mM CaCl₂. 3.1 the final LACZ concentration was 700. mu.g/mL.

Biopolymer solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. This solution was incorporated into a solution of magnetite particles having an average diameter of about 15 nm. The weight percent content of PEG-b-PLA, PCSH, and magnetite particles in this solution was about 0.071wt%, about 0.058wt%, and about 0.004wt%, respectively.

The therapeutic agent solution and the biopolymer solution are mixed and dimethyl sulfoxide or DMSO is added to form a homogeneous solution. Preparation produces particles having average diameters in the range of about 0.250 to about 1 μm, but smaller capsules can be prepared by varying process variables. In particular, the flow rate used was about 0.150 ml/h and the external voltage was about 7 kV.

Specific example 4: encapsulation of PDs Red 2NUC

The composite in this example was formed by a dual jet system as shown in figure 5. First by mixing PDS Red 2NUC with about 10mM aqueous Bis-Tris propane (containing about 1wt% isopropanol and about 2mM CaCl)₂) Mixing to prepare the core fluid solution. The final concentration of PDS Red 2NUC in the core fluid solution was about 22.5. mu.g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH in the shell fluid solution is about 0.29wt% and about 0.31wt%, respectively.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared. In particular, the core fluid flow rate and the shell fluid flow rate used were about 0.050 and about 0.300 ml/hr, respectively, and the external voltage was about 7 kV.

Capsules having PDs red 2NUC loadings in the range of about 0.01 to about 25% by weight of added polymer can be prepared by this method by adjusting the concentration of the core fluid solution.

Specific example 5: encapsulation of green fluorescent proteins

The core fluid solution is prepared by mixing green fluorescent protein or GFP with about 10mM Bis-Tris propane in water (containing about 1wt% isopropanol and about 2mM CaCl)₂) Mixing to prepare the product. The final concentration of GFP in the core fluid solution was about 30. mu.g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.32wt% and about 0.31wt%, respectively.

The preparation produced capsules having average diameters in the range of about 0.250 μm to about 1 μm, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were about 0.050 and about 0.300 ml/hr, respectively, and the external voltage was about 8 kV.

Capsules with GFP loading in the range of about 0.01 to about 20% by weight of added polymer were prepared by this method by adjusting the concentration of the core fluid solution.

Specific example 6: particles containing doxorubicin with folate functional groups

The therapeutic agent solution is prepared by dissolving doxorubicin hydrochloride or DOXO in dichloromethane. The concentration of DOXO in the therapeutic solution is about 1,000 μ g/mL.

Biopolymer solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) a folate-functionalized poly (ethylene glycol) or folate-PEG having a molecular weight of 5,000Da, and (b) a poly (caprolactone) -SH or PCSH having a molecular weight of 5,000Da and Mw/Mn = 1.5. The weight percent content of folate-PEG and PCSH is about 0.30wt% and about 0.30wt%, respectively.

The therapeutic agent solution and the biopolymer solution are mixed to form a homogeneous solution. Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate used is in the range of about 0.050 to about 0.300 ml/hr. The external voltage was about 7.5 kV.

Particles having a DOXO loading in the range of about 0.01 to about 25% by weight of added polymer can be prepared by this method by adjusting the concentration of the core fluid solution.

Specific example 7: encapsulation of doxorubicin

By mixing doxorubicin hydrochloride or DOXO in about 10mM aqueous Bis-Tris-propane (containing about 1wt% isopropanol and about 2mM CaCl)₂) To prepare a core fluid solution. The final concentration of DOXO in the core fluid solution was about 1,000 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.3wt% and about 0.3wt%, respectively.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were 0.050 and 0.300 ml/h, respectively, and the external voltage was 8.5 kV.

Capsules having a DOXO loading in the range of about 0.01 to about 23% by weight of added polymer can be prepared by this method by adjusting the concentration of the core fluid solution.

Specific example 8: encapsulation of cobalt nanoparticles

The core fluid solution is prepared by mixing cobalt nanoparticles or NP-Co in water and 10mM Bi-Tris propane in water (containing about 1wt% isopropanol and about 2mM CaCl)₂) Mixing to prepare the product. The final concentration of NP-Co in the core fluid solution is in the range of about 1,000 to about 500. mu.g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.30wt% to about 0.30wt%, respectively.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were in the range of about 0.050 to about 0.150 ml/hr, respectively, and the external voltage was about 8 kV.

Capsules having nanoparticle (Np) -Co loadings in the range of about 0.01 to about 25% by weight of added polymer can be prepared by this method by adjusting the concentration of the core fluid solution.

Specific example 9: particles comprising doxorubicin with an EGF functional group

Therapeutic agent solutions are prepared by dissolving doxorubicin hydrochloride or DOXO in dichloromethane. The concentration of DOXO in the therapeutic solution is about 1,000 μ g/mL.

Biopolymer solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) EGF-functionalized poly (ethylene glycol) or EGF-PEG, molecular weight 5,000Da, and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of EGF-PEG and PCSH is about 0.30wt% and about 0.30wt%, respectively.

The therapeutic agent solution and the biopolymer solution are mixed to form a homogeneous solution. Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate used is in the range of about 0.050 to about 0.300 ml/hr. The external voltage was about 7.5 kV.

Particles having a DOXO loading in the range of about 0.01 to about 25% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 10: luciferase as DNA-tagged particles

The therapeutic agent solution was prepared by mixing the luciferase into a 10mM Bis-Tris propane buffered water solution (containing about 1wt% isopropanol and about 2mM CaCl)₂) The preparation method is as follows. The final concentration of luciferase was about 334.5. mu.g/mL.

Biopolymer solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. This solution was incorporated into a solution of magnetite particles having an average diameter of about 15 nm. The weight percent content of PEG-b-PLA, PCSH, and magnetite particles in this solution was about 0.071wt%, about 0.058wt%, and about 0.004wt%, respectively.

The therapeutic agent solution and biopolymer solution are mixed and then dimethyl sulfoxide or DMSO is added to form a homogeneous solution. Particles having average diameters in the range of about 0.250 to about 1 μm are prepared, but smaller capsules can be prepared by varying process variables. In particular, the flow rate used was about 0.150 ml/h and the external voltage was about 7 kV.

Particles having luciferase loading in the range of about 0.01 to about 63% by weight of added polymer may be prepared by adjusting the concentration of luciferase-containing solution.

Specific example 11: particles containing TK Renilla luciferase as DNA marker

By mixing TK Renilla luciferase in about 10mM Bis-Tris propane buffered water (containing about 1wt% isopropanol and about 2mM CaC 1)₂) To prepare a therapeutic agent solution. The final concentration of TK Renilla luciferase was about 334.5. mu.g/mL.

The biopolymer solution was prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. This solution was incorporated into a solution of magnetite particles having an average diameter of about 15 nm. The weight percent content of PEG-b-PLA, PCSH, and magnetite particles in this solution was about 0.071wt%, about 0.058wt%, and about 0.004wt%, respectively.

The therapeutic agent solution and biopolymer solution were mixed, and DMSO was added to form a homogeneous solution. Particles having average diameters in the range of about 0.250 to about 1 μm are prepared, but smaller capsules can be prepared by varying process variables. In particular, the flow rate used was about 0.150 ml/h and the external voltage was about 7 kV.

Particles having a TK renilla luciferase loading in the range of about 0.01 to about 63% by weight of added polymer may be prepared by adjusting the concentration of the TK renilla luciferase-containing solution.

Specific example 12: encapsulation of cobalt nanoparticles in capsules containing EGF and a mitochondrial localization vehicle

By mixing cobalt nanoparticles or NP-Co aqueous solution and 10mM Bis-Tris propane aqueous solution (containing about 1.0wt% isopropanol and about 2mM CaCl)₂) Mixing to prepare the core fluid solution. The final concentration of NP-Co in the core fluid solution is in the range of about 1,000 to about 500. mu.g/mL.

The shell fluid solution was prepared by mixing three functionalized biopolymers in chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5; and (c) EGF-functionalized poly (ethylene glycol) or EGFPPEG, having a molecular weight of 5,000 Da. The weight percent content of PEG-b-PLA, PCSH and EGF-PEG is about 0.30wt%, about 0.30wt% and about 0.10wt%, respectively.

Buffer solutions containing plasmid subcellular localization vectors targeting mitochondria were added. The concentration of the carrier in the shell fluid solution is in the range of about 0.0 to about 1.0 wt%. DMSO was added to form a homogeneous solution.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were about 0.050 and about 0.150 ml/hr, respectively, and the external voltage was about 8 kV.

Capsules having Np-Co loadings in the range of about 0.01 to about 11% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 13: encapsulation of chlorambucil in capsules containing a CD19 functional group on the shell

By dissolving chlorambucil in about 10mM Bis-Tris propane in water (containing about 1wt% isopropanol, about 2mM CaCl)₂And DMSO) to prepare the core fluid solution. The final concentration of chlorambucil in the core fluid solution is in the range of about 2,000 to about 500 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.05wt% and about 0.05wt%, respectively. The shell fluid solution was mixed with a solution containing CD19 dissolved in a mixture of dichloromethane and polyethylene oxide (MW = 400-.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were about 0.050 and about 0.150 ml/hr, respectively, and the external voltage was about 8 kV.

Capsules having chlorambucil loadings in the range of about 0.01 to about 20.0% by weight of added polymer may be prepared by adjusting the concentration of the core fluid solution. Capsules having a CD19 load in the range of about 5 to about 100 μ g of added polymer per mg can be prepared by adjusting the concentration of the shell fluid solution.

Specific example 14: encapsulation of chlorambucil in capsules containing a CD20 functional group

By dissolving chlorambucil in about 10mM Bis-Tris propane in water (containing about 1.0wt% isopropanol, about 2.0mM CaCl₂And DMSO) to prepare the core fluid solution. The final concentration of chlorambucil in the core fluid solution is in the range of about 2,000 to about 500 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.05wt% and about 0.05wt%, respectively. The shell fluid solution was mixed with a solution containing CD20 dissolved in a mixture of dichloromethane and polyethylene oxide (MW = 400-.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were about 0.050 and about 0.150 ml/hr, respectively, and the external voltage was about 8 kV.

Capsules having chlorambucil loadings in the range of about 0.01 to about 20.0% by weight of added polymer may be prepared by adjusting the concentration of the core fluid solution. Capsules having a CD20 load in the range of about 5 to about 100 μ g of added polymer per mg can be prepared by adjusting the concentration of the shell fluid solution.

Specific example 15: encapsulation of chlorambucil and hydroxychloroquine sulfate in capsules containing CD19 and CD20 functional groups

By passingChlorambucil and hydroxychloroquine sulfate or HCQ were dissolved in about 10mM Bis-Tris propane in water (containing about 1.0wt% isopropanol, 2mM CaCl₂And DMSO) to prepare the core fluid solution. The final concentrations of chlorambucil and HCQ in the core fluid solution are in the range of about 2,000 to about 500 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percentage content of PEG-b-PLA and PCSH is 0.05wt% and 0.05wt%, respectively. The shell fluid solution was mixed with a solution containing CD19 and CD20 dissolved in a mixture of dichloromethane and polyethylene oxide (MW = 400-.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were about 0.050 and about 0.300 ml/hr, respectively, and the external voltage was about 8 kV.

Capsules having a loading of chlorambucil and HCQ in the range of about 0.01 to about 20.0% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution. Capsules having CD19 and CD20 loadings in the range of about 5 to about 100 μ g of added polymer per mg can be prepared by adjusting the concentration of the shell fluid solution.

Specific example 16: encapsulation of chlorambucil in capsules containing a CD19 functional group and a Golgi complex localization vehicle

By dissolving chlorambucil in about 10mM Bis-Tris propane in water (containing about 1wt% isopropanol, about 2mM CaCl)₂And DMSO) to prepare the core fluid solution. The final concentration of chlorambucil in the core fluid solution ranges from about 2,000 to about 500 μ g/mL.

The shell fluid solution was prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.05wt% and about 0.05wt%, respectively. The shell fluid solution was mixed with a solution containing CD19 dissolved in a mixture of dichloromethane and polyethylene oxide (MW = 400-.

Buffer solutions containing plasmid subcellular localization vectors targeting the golgi complex were added. The concentration of the carrier in the shell fluid solution is in the range of about 0 to about 1 wt%. DMSO was added to form a homogeneous solution.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were about 0.050 and about 0.150 ml/hr, respectively, and the external voltage was about 8 kV.

Capsules having chlorambucil loadings in the range of about 0.01 to about 20.0% by weight of added polymer may be prepared by adjusting the concentration of the core fluid solution. Capsules having a CD19 load in the range of about 5 to about 100 μ g of added polymer per mg can be prepared by adjusting the concentration of the shell fluid solution.

Specific example 17: particles containing iodine-125 and EGF functional groups

A therapeutic agent solution was prepared by dissolving iodine-125 in dichloromethane. The concentration of iodine-125 in the therapeutic solution is about 1,000 μ g/mL.

Biopolymer solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) EGF-functionalized poly (ethylene glycol) or EGF-PEG, molecular weight 5,000Da, and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of EGF-PEG and PCSH is about 0.30wt% and about 0.30wt%, respectively.

The therapeutic agent solution and the biopolymer solution are mixed to form a homogeneous solution. Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate used is in the range of about 0.050 to about 0.300 ml/hr. The external voltage was about 7.5 kV.

Particles having iodine-125 loadings in the range of about 0.01 to about 25% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 18: encapsulation of chlorambucil in capsules containing Ga/Fe nanoparticles and epidermal growth factor receptor

By dissolving chlorambucil in 10mM Bis-Tris propane aqueous solution (containing about 1.0wt% isopropanol, about 2.0mM CaCl₂And DMSO) to prepare the core fluid solution. The final concentration of chlorambucil in the core fluid solution ranges from about 2,000 to about 500 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers in chloroform: (a) epidermal growth factor-functionalized poly (ethylene glycol) or EGF-PEG with a molecular weight of 5,000Da, and (b) poly (caprolactone) -SH or PCSH with a molecular weight of 5,000Da and Mw/Mn = 1.5. The solution was doped with Fe having an average diameter of about 15nm₃O₄And Ga₂O₃A solution of nanoparticles. EGF-PEG, PCSH, Fe in a shell fluid solution₃O₄And Ga₂O₃Are present in an amount of about 0.071wt%, about 0.058wt%, about 0.004wt%, and about 0.004wt%, respectively.

Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate used is in the range of about 0.050 to about 0.300 ml/hr. The external voltage was about 7.5 kV.

Particles having chlorambucil loadings in the range of about 0.01 to about 25.0% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 19: encapsulation of chlorambucil in capsules containing Ga/B/Fe nanoparticles and epidermal growth factor receptor

By dissolving chlorambucil in about 10.0mM Bis-Tris propane in water (containing about 1.0wt% isopropanol, about 2.0mM CaCl₂And DMSO) to prepare the core fluid solution. The final concentration of chlorambucil in the core fluid solution is in the range of about 2,000 to about 500 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers in chloroform: (a) epidermal growth factor-functionalized poly (ethylene glycol) or EGF-PEG with a molecular weight of 5,000Da, and (b) poly (caprolactone) -SH or PCSH with a molecular weight of 5,000Da and Mw/Mn = 1.5. The solution was doped with Fe having an average diameter of about 15nm₃O₄、Ga₂O₃And B₂O₃A solution of nanoparticles. EGF-PEG, PCSH, Fe in a shell fluid solution₃O₄、Ga₂O₃And B₂O₃Are present in an amount of about 0.071wt%, about 0.058wt%, about 0.004wt%, and about 0.004wt%, respectively.

Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate used is in the range of about 0.050 to about 0.300 ml/hr. The external voltage was about 7.5 kV.

Particles having chlorambucil loadings in the range of about 0.01 to about 25% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 20: paclitaxel-containing particles with folate functionality

Therapeutic agent solutions were prepared by dissolving paclitaxel in dichloromethane. The concentration of paclitaxel in the therapeutic solution is about 1,000 μ g/mL.

Biopolymer solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) a folate-functionalized poly (ethylene glycol) or folate-PEG having a molecular weight of 5,000Da, and (b) a poly (caprolactone) -SH or PCSH having a molecular weight of 5,000Da and Mw/Mn = 1.5. The weight percent content of folate-PEG and PCSH is about 0.30wt% and about 0.30wt%, respectively.

The therapeutic agent solution and the biopolymer solution are mixed to form a homogeneous solution. Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate used is in the range of about 0.050 to about 0.300 ml/hr. The external voltage was about 7.5 kV.

Particles having a paclitaxel loading in the range of about 0.01 to about 25% by weight of the added polymer are prepared by adjusting the concentration of the core fluid solution.

Specific example 21: paclitaxel-containing capsules with folic acid functionality

The core fluid solution was prepared by mixing paclitaxel into DMSO and about 10wt% gamma-cyclodextrin in a 10mM aqueous Bis-Tris propane solution (containing about 1wt% isopropanol and about 2mM CaCl)₂) In (1). The final concentration of paclitaxel in this core fluid solution was about 20 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers in chloroform: (a) folic acid functionalized poly (ethylene glycol) or folic acid-PEG, molar weight 5,000 Da; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5000Da and Mw/Mn = 1.5. The weight percent content of each of folate-PEG and PCSH is about 0.30 wt%.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.300 ml/hr, respectively, and the external voltage is about 7.5 kV.

Capsules loaded with paclitaxel in the range of about 0.01 to about 0.2% by weight of the added polymer are prepared by adjusting the concentration of the core fluid solution.

Specific example 22: particles of paclitaxel

Therapeutic agent solutions are prepared by mixing paclitaxel into dichloromethane. The concentration of paclitaxel in the therapeutic solution is about 1,000 μ g/mL.

Biopolymer solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.3wt% and about 0.3wt%, respectively.

The therapeutic agent solution and the biopolymer solution are mixed to form a homogeneous solution. Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate is in the range of about 0.050 to about 0.300 ml/hr and the external voltage is about 7.5 kV.

Particles having a paclitaxel loading in the range of about 0.01 to about 25% by weight of the added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 23: capsule containing paclitaxel

The core fluid solution was prepared by mixing paclitaxel into DMSO and about 10wt% gamma-cyclodextrin. The 10wt% gamma-cyclodextrin was in an aqueous solution of about 10mM Bis-Tris propane (containing about 1wt% isopropanol and about 2mM CaCl)₂) In (1). The final concentration of paclitaxel in this core fluid solution was about 20 μ g/mL.

The shell fluid solution was prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH, respectively, is about 0.3 wt%.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.300 ml/hr, respectively, and the external voltage is about 7 kV.

Capsules having a paclitaxel loading in the range of about 0.01 to about 0.2% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 24: encapsulation of gold nanoparticles

By mixing an aqueous solution of gold nanoparticles or NP-Au with an aqueous solution of 10mM Bis-Tris-propane (containing about 1wt% isopropanol and about 2mM CaCl)₂) Mixing to prepare the core fluid solution. The final concentration of NP-Au in the core fluid solution is in the range of about 1,000 to about 500. mu.g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.30wt% and about 0.30wt%, respectively.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.150 ml/hr, respectively, and the external voltage is about 8 kV.

Capsules having gold nanoparticle loadings in the range of about 0.01 to about 11% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 25: encapsulation of silver nanoparticles

By mixing an aqueous solution of silver nanoparticles or NP-Ag with an aqueous 10mM Bis-Tris-propane solution (containing about 1wt% isopropanol and about 2mM CaC 1)₂) Mixing to prepare the core fluid solution. The final concentration of NP-Ag in the core fluid solution ranges from about 1,000 to about 500. mu.g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.30wt% and about 0.30wt%, respectively.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.150 ml/hr, respectively, and the external voltage is about 8 kV.

Capsules having silver nanoparticle loadings in the range of about 0.01 to about 11% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 26: encapsulation of palladium nanoparticles

By mixing an aqueous solution of palladium nanoparticles or Np-Pd and an aqueous solution of about 10mM Bis-Tris propane (containing about 1wt% isopropanol and about 2mM CaCl)₂) Mixing to prepare the core fluid solution. The final concentration of NP-Pd in the core fluid solution ranges from about 1,000 to about 500. mu.g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.30wt% and about 0.30wt%, respectively.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.150 ml/hr, respectively, and the external voltage is about 8 kV.

Capsules having palladium nanoparticle loadings in the range of about 0.01 to about 11% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 27: particles with paclitaxel and estradiol functionalities

The therapeutic agent solution is prepared by mixing paclitaxel into dichloromethane. The concentration of paclitaxel in the therapeutic solution is about 1,000 μ g/mL.

Biopolymer solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) estradiol functionalized poly (ethylene glycol) or EST-PEG with a molecular weight of 5,000 Da; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of EST-PEG and PCSH, respectively, is about 0.30 wt%.

The therapeutic agent solution and the biopolymer solution are mixed to form a homogeneous solution. Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate is in the range of about 0.050 to about 0.300 ml/hr and the external voltage is about 7.5 kV.

Particles having a paclitaxel loading in the range of about 0.01 to about 25% by weight of the added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 28: capsule with paclitaxel and estradiol functional groups

The core fluid solution was prepared by mixing paclitaxel into DMSO and about 10wt% gamma-cyclodextrin in a 10mM aqueous Bis-Tris propane solution (containing about 1wt% isopropanol and 2mM CaCl)₂) In (1). The final concentration of paclitaxel in this core fluid solution was about 20 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers in chloroform: (a) estradiol functionalized poly (ethylene glycol) or EST-PEG with a molar weight of 5,000 Da; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of EST-PEG and PCSH, respectively, is about 0.30 wt%.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.150 ml/hr, respectively, and the external voltage is about 8 kV.

Specific example 29: particles with paclitaxel and epidermal growth factor groups

Therapeutic agent solutions were prepared by dissolving paclitaxel in dichloromethane. The concentration of paclitaxel in the therapeutic solution is about 1,000 μ g/mL.

Biopolymer solutions were prepared by mixing three functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA having a molecular weight of 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2, (b) poly (caprolactone) -SH or PCSH having a molecular weight of 5,000Da and Mw/Mn = 1.5; and (c) epidermal growth factor-functionalized poly (ethylene glycol) or EGF-PEG, having a molecular weight of 5,000 Da. The weight percent content of PEG-b-PLA, PCSH and EGF-PEG is about 0.30wt%, about 0.30wt% and about 0.10wt%, respectively.

The therapeutic agent solution and the biopolymer solution are mixed to form a homogeneous solution. Particles having average diameters in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the flow rate is in the range of about 0.050 to about 0.300 ml/hr. The external voltage was about 7.5 kV.

Particles having a paclitaxel loading in the range of about 0.01 to about 25% by weight of the added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 30: capsule with paclitaxel and epidermal growth factor group

The core fluid solution was prepared by mixing paclitaxel into DMSO and about 10wt% gamma-cyclodextrin in a 10mM aqueous Bis-Tris propane solution (containing about 1wt% isopropanol and 2mM CaCl)₂) In (1). The final concentration of paclitaxel in this core fluid solution was about 20 μ g/mL.

Shell fluid solutions were prepared by mixing three functionalized biopolymers in chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5; and (c) epidermal growth factor-functionalized poly (ethylene glycol) or EGF-PEG, having a molecular weight of 5,000 Da. The weight percent content of PEG-b-PLA, PCSH and EGF-PEG is about 0.30wt%, about 0.30wt% and about 0.10wt%, respectively.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.300 ml/hr, respectively, and the external voltage is about 7.5 kV.

Capsules having a paclitaxel loading in the range of about 0.01 to about 0.2% by weight of added polymer are prepared by adjusting the concentration of the core fluid solution.

Specific example 31: encapsulation of pCMV-Luc plasmid

By mixing a buffer solution containing a pCMV-Luc plasmid containing the Cytomegalovirus (CMV) promoter of firefly luciferase pcDNA3 inserted countercurrently into pGL 2-basic vector plasmid and a 10mM Bis-Tris propane aqueous solution containing about 1wt% isopropanol and about 2mM CaCl₂) Mixing to prepare the core fluid solution. The final concentration of pCMV-Luc in the core fluid solution is in the range of about 1 to about 1000 μ g/mL.

Shell fluid solutions were prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The weight percent content of PEG-b-PLA and PCSH is about 0.30wt% and about 0.30wt%, respectively.

Capsules having an average diameter in the range of about 0.250 μm to about 1 μm are prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.150 ml/hr, respectively, and the external voltage is about 8 kV.

Capsules having pCMV-Luc loadings in the range of about 0.01 to about 11% of the weight of polymer added can be prepared by adjusting the concentration of the core fluid solution.

Example 32 encapsulation of 425TNF- α DNA and bovine serum Albumin rhodamine conjugate

By placing 425TNF- α DNA and bovine serum albumin rhodamine conjugate or BSA-rhodamine in about 10mM Bis-Tris propane aqueous solution (containing about 1wt% isopropanol and about 2mM CaCl₂) 425TNF- α DNA and BSA-rhodamine at final concentrations of about 12.2. mu.g/mL and about 200. mu.g/mL, respectively, in the core fluid solution.

The shell fluid solution was prepared by mixing EGF or F-PEG-EGF grafted to fluorescein-labeled poly (ethylene glycol) -NHS biopolymer, with a molecular weight of 3,400 Da. The amount of F-PEG-EGF in the shell fluid solution was about 0.317 wt%.

Capsules with an average diameter of about 0.41 μm were prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate used were about.05 and about 0.30 ml/hr, respectively, and the voltage used was about 6.5 kV.

Capsules having a 425TNF- α DNA load in the range of about 0.01 to about 30% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

Specific example 33: encapsulation of temozolomide

The core fluid solution was prepared by mixing temozolomide or TMZ into about 0.1M acetate buffer solution. The final concentration of TMZ in the core fluid solution was about 10 μ M.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COON-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The PEG-b-PLA and PCSH content in the shell fluid solution was about 0.080wt%, about 0.086wt%, respectively.

Capsules with an average diameter of about 1.3 μm were prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate were about 0.05 and about 0.30 ml/hr, respectively, and the applied voltage was about 8.0 kV.

Capsules having temozolomide loadings in the range of about 0.01 to about 2.4% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

EXAMPLE 34 encapsulation of TNF- α protein and bovine serum albumin fluorescein conjugate

By placing TNF- α protein and bovine serum albumin fluorescein conjugate or BSA-fluorescer 594 in about 10mM Bis-Tris propane in water (containing about 1wt% isopropanol and about 2mM CaCl)₂) The final concentrations of TNF- α protein and the BSA-fluorescing agent in the core fluid solution were about 345. mu.g/mL and about 200. mu.g/mL, respectively.

Shell fluid solutions were prepared by mixing two functionalized biopolymers into chloroform: (a) COOH-poly (ethylene glycol) -b-polylactide or PEG-b-PLA, molecular weight 2000-b-1940Da, using the same nomenclature, respectively, and Mw/Mn =1.2; and (b) poly (caprolactone) -SH or PCSH, molecular weight 5,000Da and Mw/Mn = 1.5. The shell fluid solution was incorporated into a solution of magnetite particles having an average diameter of about 15 nm. The content of PEG-bPLA, PCSH and magnetite particles in the shell fluid solution was about 0.080wt%, about 0.086wt% and about 0.006, respectively.

Capsules having average diameters of about 0.225 μm and about 0.550 μm were prepared, but smaller capsules can be prepared by adjusting process variables. In particular, the core fluid flow rate and the shell fluid flow rate are about 0.050 and about 0.300 ml/hr, respectively, and the applied voltage is about 7.0 kV.

Capsules having a loading of TNF- α protein in the range of about 0.01 to about 23% by weight of added polymer can be prepared by adjusting the concentration of the core fluid solution.

The examples given above are merely illustrative and do not represent an exhaustive list of all possible embodiments, applications or modifications of the invention. Thus, various modifications and variations of the described methods and systems of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. While the invention has been described in conjunction with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in polymer science, molecular biology or related fields are intended to be within the scope of the following claims.

The disclosures of all references and publications cited above are expressly incorporated by reference in their entirety to the same extent as if each were incorporated by reference.

Claims (12)

1. A capsule prepared at least in part by using an electrical potential and a hollow tube formed at least in part by providing a fluid to an inner wall, an outer wall and a top at a flow rate in a range of about 0.005 ml/hr to about 5 ml/hr, the capsule comprising:

a core region formed at least partially through the inner wall;

a shell region formed at least in part via the outer wall;

the core region and shell region are exposed to the electrical potential so as to form an electrically charged jet at the top of the hollow tube;

the shell region comprises pores having a size in a range of about 0.5 nanometers to about 2.0 nanometers;

at least one embedded and/or encapsulated agent; and

functional groups dispersed on the surface of the shell region.

2. The capsule of claim 1, wherein the encapsulated and/or embedded agent is at least one of a therapeutic agent and an imaging agent.

3. The capsule of claim 1, wherein magnetic nanoparticles are dispersed in the shell region.

4. The capsule of claim 1, wherein the functional group has an affinity to chemically or physically attach to a target cell.

5. The capsule of claim 4, wherein the target cell is a malignant cancer cell selected from the group consisting of: gliomas, lymphomas, leukemias, carcinomas, sarcomas, mesotheliomas, gliomas, and germ cell tumors.

6. The capsule of claim 4, wherein the target cell is a malignant cancer cell selected from the group consisting of: breast, prostate, pancreatic and choriocarcinoma.

7. The capsule of claim 2, wherein the therapeutic agent is at least one of a chemotherapeutic agent, a radioisotope, and a chemotherapeutic enhancer.

8. The capsule of claim 7, wherein the therapeutic agent is a compound selected from the group consisting of: anti-inflammatory compounds, anti-allergic agents, glucocorticoids, antibiotics, antifungal agents, antiviral agents, mucolytics, antiseptics, vasoconstrictors, wound healing agents, local anesthetics, peptides and proteins.

9. The capsule of claim 7, wherein the therapeutic agent is an anti-infective agent.

10. The capsule of claim 1, wherein the functional group is capable of targeting a subcellular target.

11. The capsule of claim 10, wherein the subcellular target is one or more components selected from the group consisting of: endoplasmic reticulum, mitochondria, golgi apparatus, vacuole, nucleus, centromere, cilia, lysosomes, melanosomes, myofibrils, peroxisomes, actin, tubulin, plasma membrane, ribosomes, and vesicles.

12. The capsule of claim 10, wherein the subcellular target is one or more components selected from the group consisting of: acrosomes, glyoxylate cycle bodies, and nucleoli.