METHODS FOR PRODUCING MULTILAYERED PARTICLES, FIBERS AND SPRAYS AND METHODS FOR ADMINISTERING THE SAME
Capsules and particles with at least one encapsulated and/or entrapped agent, such as therapeutic agent, imaging agents, and other constituents may be produced by electrohydrodynamic processes. More particularly, the agent encapsulated in a vehicle, capsule, particle, vector, or carrier may maximize treatment and/or imaging of malignant cancers while minimizing the adverse effects of treatment and/or imaging.
This application claims priority to and the benefit of U.S. Provisional Application Ser. No. 60/746,311, filed May 3, 2006, and U.S. Provisional Application Ser. No. 60/886,225, filed Jan. 23, 2007, the disclosures of which are expressly incorporated herein by reference in their entirety.
BACKGROUND1. Field of Invention
The invention relates to systems and methods for producing capsules and particles with at least one encapsulated and/or entrapped agent, such as therapeutic agent, imaging agents, and other constituents. More particularly, the agent encapsulated in a vehicle, capsule, particle, vector, or carrier may maximize treatment and/or imaging of malignancy while minimizing the adverse effects of treatment and/or imaging.
2. Related Art
Cancer is a class of diseases or disorders characterized by uncontrolled division of cells and the ability of these to spread, either by direct growth into adjacent tissue through invasion, or by implantation into distant sites by metastasis. Cancer may affect people at all ages, bat risk tends to increase with age. It is one of the principal causes of death in developed countries.
For example, according to the National Cancer Institute, breast cancer affected between 12 and 13 per every 10,000 women in 2003. Although imaging and early diagnostic tools have been improving over the past two decades, current breast cancer early detection is far from infallible, especially when using mammograms in younger women. While magnetic resonance imaging (MRI and ultrasound) and laser-based imaging techniques have been used or evaluated, the issue for best possible contrast between healthy and cancerous tissue for any imagining technique is a longstanding one.
A current challenge facing scientists is determining how to design a therapeutic or an imaging agent and its vehicle, vector or carrier in order to maximize treatment and imaging of malignant cancers in patients while minimizing the adverse effects of treatment. Moreover, the selective delivery of therapeutic agents to a desired part of the body is also a nontrivial issue. Current treatments may lead to insufficient tumor distribution or therapeutic agents and often cause adverse effects on patients. Systemic injections of therapeutic agents carry consequences associated with their nonspecific dispersion in the body and have a limited therapeutic agent distribution throughout the targeted malignancy. One approach to overcome these short comings is to design an effective therapeutic or imaging agent delivery vehicle by making vesicles or capsules containing the desired therapeutic agent.
Formation of vesicles or capsules that are small enough to be delivered into the human body by means of inhalation, injection or permeation through the skin has received significant attention. The outer skin or shell of the vesicles may be chemically functionalized with receptors and other species to selectively target certain organs. There are several methodologies available, such as electrospray and two-capillary jet systems to fabricate small vesicles.
Emulsion-polymerization technologies, such as DC coaxial electrospray, AC coaxial electrospray, and electrohydrodynamics (EHD) are well known methods that may produce micro-sized capsules. In general, a solution containing the target compound or compounds to be encapsulated, also called “encapsulates,” is emulsified into another fluid to solution having at least one substance capable of forming a shell or envelope around the encapsulate dispensed droplets. Although emulsion-polymerization methods are relatively scalable, there are several limitations associated with the methods, such as the inability to encapsulate the targets in a quantitative manner, and the high-shear production of emulsions may compromise the integrity of mechanically delicate encapsulates, such as biological constituents such as proteins, genetic material, and other molecules of biological origin.
Coaxial electrospray based on application of DC electrical potentials between a coaxial capillary fixture delivering the encapsulate-containing core liquid and a shell-forming liquid precursor, and a collector surface or counter-electrode, has the ability to produce vesicles in the micrometer and sub-micrometer range. Although DC coaxial electrospray may be relatively gentle to biological encapsulates, its major limitation is that it lacks simplicity in equipment, design, scalability, and thus cost effectiveness.
As an alternative to DC based electrospray, AC electrospray may be employed. For example, AC electrospray may be used to produce encapsulates entrapped within a bioabsorbable biopolymeric matrix, such as polylactic acid. AC electrospray yields essentially an electrically neutral electrospray. While there are advantages associated with charge neutrality, such as decreased ability of the particles or electrospray droplets to absorb indiscriminately over non-targeted surfaces, and avoidance of a potential charge buildup problem. One disadvantage of AC electrospray is that it produces undesirablely large particles having sizes well above one micron. For many medical applications, such as penetration of the blood brain barrier. AC electrospray derived particles are unacceptably large.
In addition to electrospray methodologies, coaxial liquid jet system combined with sol-gel chemistry, such as EHD, may be employed to fabricate vesicles or capsules. Use of coaxial, two-capillary coaxial arrangements to simultaneously deliver two fluids in the presence of electric field gradients are well know in the art.
Briefly, in this method, the chemistry and physical properties of the two fluids and the values of variables such as electric field strength and flow rates of the two fluids may determine the structure of matter collected onto a collector electrode, which may be located at a distance from the exit region of the two-capillary coaxial arrangement. At the exit region, the compound two-fluid structure may form an electrified meniscus that may adopt various shapes, such as a Taylor cone.
Laminar flows may be non-turbulent, which may minimize mixing between flowing fluid layers. Thus, since the two fluids may not mix to the point of forming a single fluid phase, the thinning electrified two-fluid jet may enter into a chaotic path resembling whipping phenomena. At a point along its path toward a collection zone or collector body 112, the compound two-fluid jet may experience an electrical charge oscillatory phenomenon known as Rayleigh instability. This may cause the compound two-fluid jet to no longer experience progressive thinning, but an oscillatory thinning and thickening regime which may eventually lead to jet breakup into a droplet-in-droplet regime, or compound electrospray regime.
The chemical and physical properties of the two fluids may be controlled to produce a variety of structures collected at the collection zone or collector electrode 112. For example, if fluid 106 yields, a solid structure through solvent evaporation and precipitation of a solid phase, fluid 104 may be encapsulated into structures such as hollow fibers, hollow beaded fibers or capsules, for example. Alternatively, the chemical and physical properties of the two liquids may be adjusted to cause no solidification of fluid 104 and fluid 106, solidification of one of the two fluids, or solidification of both fluids, during the time of travel of the compound charged structures from the two-fluid electrified meniscus to the collection zone 112.
Referring to
Although coaxial two-capillary systems may be employed to produce the core-shell structures, described above, there are several disadvantages associated with the conventional systems. In particular, when a direct, parallel scale-up of the process to increase process throughput is attempted a micro-fabrication problem occurs. For instance, a typical range of internal diameters for the inner and outer capillaries are about 0.1 to about 0.3 mm and about 0.3 to about 1.0 mm, respectively. In order to build an instrument consisting of many such coaxial, two-capillary fixtures for scaled up production of a desired core-shell structure, it is necessary to produce each individual fixture with inner and outer capillaries aligned as close to coaxial as possible, and also with high reproducibility in their diameters. With modern micro-fabrication techniques such challenge may be met, however, these techniques are very complex and not cost effective.
In particular, a conventional way to manage fluid flow through many orifices, capillaries, conduits or two-capillary coaxial fixtures is by using one means for forcing flow through all same fluid orifices, capillaries, conduits or two-capillary coaxial fixtures, not by controlling the fluid flow rate through each individual fluid flow path. This is the reason why, for example, fabricating a parallel scaled up production of a desired core-shell structure is difficult and expensive. If there is variability in diameter from inner or outer capillary of one two-capillary coaxial fixture to another in excess of about 2% or 3%, it is not possible to produce a desired core-shell structure without the occurrence of undesirable structures. With such prior art two-capillary fixture, small differences in the overall pressure drop profiles of the inner and outer capillaries also causes undesirable effects.
SUMMARY OF THE INVENTIONThe invention satisfies the above needs by providing systems and methods for fabricating capsules with entrapped or encapsulated therapeutic agents and/or imaging agents. More particularly, the systems and methods of the invention permits a wider range of capsule size, keeps chemical interactions leading to degradation of therapeutic agents and/or imaging agents to sufficiently low and acceptable levels, involves less manufacturing steps, and is more cost effective than conventional methods.
According to one aspect of the invention, a method for producing a capsule having at least one encapsulated agent is provided. The method may include supply a core fluid to an interior of a hollow tube, supply a shell fluid flow, generating a shell fluid flow to an external wall of the hollow tube, subjecting the core fluid and the shell fluid to an electric potential so as to form a two-fluid electrified jet at an apex of the hollow tube, and forming a capsule having a core region and a shell region at the apex of the hollow tube. Moreover, the method may also include supplying a collector electrode and depositing the formed capsules onto a surface of the collector electrode.
In another aspect, the methodology of the invention may include dispersing at least one species of a magnetite particle within the shell region of the capsule. The at least one species of magnetite particle may have a size in the range of about 1 nm to about 300 nm. The magnetite particle species may be Fe3O4.
In yet another aspect, the methodology may include dispersing at least one species of a photonic sensitive nanoparticle within the shell region of the capsule. The photosensitive nanoparticle species may have a size in the range of about 1 nm to about 50 nm. The species of photonic sensitive nanoparticle may include silver, gold, palladium, and any combination thereof.
In a further aspect, the core fluid may be supplied at a flow rate in the range of about 0.05 mL/hour to about 5 mL/hour, and specifically, at a flow rate of about 0.025 mL/h. The shell fluid may be supplied at a flow rate in the range of about 0.05 mL/hour to about 5 mL/hour, and particularly, at a flow rate of about 0.75 mL/h. The positive electrical bias may have a voltage of about 5 kV to about 18 kV. The collector electrode may be at ground electrical potential.
In one aspect, the methodology may include dispersing a functional group on the outer perimeter of the shell of the capsule. The function group may be entity such as hydroxyl groups, amino groups, carboxyl groups, carboxylic acid anhydride groups, mercapto groups, hydrosilicon groups, thio groups, carboxylic acid groups, amine groups, and any combination thereof. Furthermore, the functional group may be capable of chemically or physically attaching to pancreatic cancer cells. The functional group may be one or more compounds such as kinase receptors, fibroblast growth factor receptor, EGF, TGF, VEGF-A, urokinase receptor, interleukin-4 receptor, retinoic acid receptor, heparin-binding EGF-like growth factor, HB-EGF, amphiregulin, epireguin, neuregulins, and functional equivalents thereof. The functional group may have a content in the range of about 0 wt % to about 1 wt %.
In a specific aspect, the functional group may be capable of chemically or physically attaching to glioma cells and may be epidermal growth factor and a functional equivalent thereof. The functional group may have a content on the capsule in the range of about 0 wt % to about 0.02 wt %.
In another specific aspect, the function group may be capable of chemically or physically attaching to breast cancer cells. The functional group may be estrogen and functional equivalents thereof. Additionally, the functional group may have a content on the capsule in the range of about 0.02 wt % to about 0.4 wt %.
In yet another specific aspect, the functional group may be capable of chemically or physically attaching to at least one of lymphoma, myeloma and leukemia cancer cells. The functional group may include one or more of VEGR-2, tostumomab, antibodies capable of binding to a CD20 receptor, CD5, CD7, CD13, CD19, CD22, CD33, CD52, CD61, antimyeloperoxidase, and functional equivalents thereof. The function group may have a content on the shell of the capsule in the range of about 0.0 wt % to about 1.0 wt %.
According to one aspect of the invention, the shell fluid may be at least one biocompatible polymer. The biocompatible polymer may include one or more of poly(lactic acid), poly(lactic-co-glycolic acid), citosan, alkyl metacrylates, polycaprolactore, starch, polyethylene glycol, and copolymers resulting from combination thereof.
In a particular, aspect, the core fluid may include one or more of a therapeutic agent and an imaging agent. The therapeutic agent may include one or more materials such as nucleic acid, double stranded DNA, single stranded DNA, double stranded RNA, single stranded RNA, peptide nucleic acid (PNA), antisense DNA, antisense RNA, small interfering RNA, proteins, lipids, carbohydrates, and combinations thereof. Furthermore, the therapeutic agent is at least one abiological material. The imaging agent is suitable for detecting a malignant cancer in a patient.
In another aspect, the therapeutic agent may be suitable for a treatment a patient afflicted with a condition. The patient may be afflicted with at least one condition such as breast cancer, glioma, lymphoma, leukemia, prostate cancer, pancreatic cancer, carcinoma, sarcoma, mesothelioma, glioma, germinoma, and choriocarcinoma.
According to one aspect, a capsule including at least one encapsulated agent may be produced by the methodology of the invention, described above. In particular, the encapsulated agent may contain at least one of a therapeutic agent and an imaging agent. The capsule may include a shell region and a core region. Additionally, magnetic nanoparticles may be dispersed in the shell region. Moreover, the shell region of the capsule may include pores. The pores may have a size in the range of about 0.5 nm to about 2.0 nm.
In a specific aspect, functional groups may be dispersed on an outer surface of the shell region. The functional group may have an affinity to attach chemically or physically to a target cell. The target cell may be a malignant cancer cell such as breast cancer, glioma, lymphoma, leukemia, prostate cancer, pancreatic cancer, carcinoma, sarcoma, mesothelioma, glioma, germinoma, and choriocarcinoma.
In a more specific aspect, the therapeutic agent may be at least one of a chemotherapeutic agent, radioactive isotope, and a chemotherapeutic enhancing drug. The therapeutic agent may be a compound such as anti-inflammatory compounds, anti-allergics, glucocorticoids, anti-infective agents, antibiotics, antifungals, antivirals, mucolytics, antiseptics, vasoconstrictors, wound healing agents, local anaesthetics, peptides, and proteins.
Additional features, advantages, and embodiments of the invention may be set forth or apparent from consideration of the following detailed description, drawings, and claims. Moreover, 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.
BRIEF DESCRIPTION O THE DRAWINGSThe accompanying drawings, which are included to provide a further understanding of the invention, are incorporated in and constitute a part of this specification; illustrate embodiments of the invention and together with the detailed description serve to explain the principles f the invention. No attempt if made to show structural details of the invention in more detail than may be necessary for a fundamental understanding of the invention and various ways in which it may be practiced.
It is understood that the invention is not known to the particular methodology, protocols, end reagents, etc., described herein, as these may vary as the skilled artisan will recognize. It is also to be understood that the terminology used herein is used for the purpose of describing particular embodiments only, and is not intended to limit the scope of the invention. It also is be noted that as used herein, and in the appended claims, the singular forms “a,” “an,” and “the” include the plural reference unless the context clearly dictates otherwise. This, for example, a reference to “a capsule” is a transference to one or more capsules and equivalents thereof known m those skilled in the art.
Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the invention pertains. The embodiments of the invention and the various features and advantageous details thereof are explained more fully with reference to the non-limiting embodiments and/or 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 wads other embodiments as the skilled artisan would recognize, even if not explicitly stated herein.
Any numerical values recited herein include all values from the lower value to the upper value in increments of one unit provided that there is a separation of at least two units between any lower value and any higher value. As an example, if it is stated that the concentration of a component or value of a process variable such as, for example, size, temperature, pressure, time and the like, is, for example, from 1 to 90, specifically from 20 to 80, more specifically 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 which are less than one, one unit is considered to be 0.0001, 0.001,0,01 or 0.1 as appropriate. These are only examples of what is specifically intended 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.
Moreover, provided immediately below is a “Definition” section, where certain terms related to the invention are defined specifically. Particular 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 invention. All references referred to herein are incorporated by reference herein in their entirety.
DEFINITIONSBSA is bovine serum albumin
CED is convection enhanced delivery
CFM is confocal fluorescence microscopy
EHD is electrohydrodynamics
PBS is phosphate-buffer saline
PEG is Poly(ethylene glycol)
PLA is poly(lactic acid)
PLC 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 material or compound which, when administered to an organism (human or animal) induces a desired pharmacologic effect. Included are derivatives and analogs of those compounds or classes of compounds specifically mentioned that also induce the desired pharmacologic effect. In particular, the therapeutic agent may encompass a single biological or abiological chemical compound, or to a combination of biological and abiological compounds that may be required to cause a desirable therapeutic effect.
By “pharmaceutically acceptable carrier” is meant a material or materials that are suitable for drug administration and not biologically or otherwise undesirable, i.e., that may be administered to an individual along with an active agent without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical formulation in which it is contained.
Similarly, a “pharmacologically acceptable” salt, ester or other derivative of an active agent as provided herein ia a salt, ester or other derivative that is not biologically or otherwise undesirable.
By the terms “effective amount” or “therapeutically effective amount” of an agent as provided herein are meant a nontoxic 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. Thus, it is not possible to specify an exact “effective amount.” However, an appropriate “effective” amount in any individual case may be determined by one of ordinary skill in the art using only routine experimentation.
The terms “treating” and “treatment” as used herein refer to reduction in severity and/or frequency of symptoms, elimination of symptoms and/or underlying cause, prevention of the occurrence of symptoms and/or their underlying cause, and improvement or remediation of damage. Thus, for example, the present method of “treating” individuals with cancer, as the term “treating” is used herein, encompasses treatment of cancer in a clinically symptomatic individual.
The terms “condition,” “disease” and “disorder” are used interchangeably herein as referring to a physiological state that can be detected, prevented or treated by administration of a therapeutic agent as described herein. Exemplary diseases and conditions may include, but not limited to, cancer such as breast cancer, glioma, pancreatic cancer, leukemia, and, lymphoma.
The term “patient” as in treatment of “a patient” refers to a mammalian individual afflicted with or prone to a condition, disease or disorder as specified herein, and includes both humans and animals.
The term “nucleic acid,” as used herein may include an oligonucleotide, nucleotide, or polynucleotide, and fragments thereof, and to DNA or RNA of genomic or synthetic origin which may be single- or double-stranded, and represent the sense or antisense strand, to peptide nucleic acid (PNA), to small interfering RNA (siRNA) molecules, or to any DMA-like or RNA-like material, natural or synthetic in origin.
The term “transfection,” as used herein includes the process of introducing 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 changes a recipient cell. It may occur under natural or artificial conditions using various methods well known in the art. Transformation may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. The method is selected based on the type of host cell being transformed and may include, but is not limited to, viral infection, electroporation, heat shock, and lipofection.
Antisense gene: an antisense gene is constructed by reversing the orientation of the gene with respect to its promoter so that the antisense strand is transcribed.
Antisense RNA: an RNA molecule complementary to a particular 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 nucleic acid molecule that possesses functional or structural characteristics that is substantially similar to a heterologous protein, polypeptide, enzyme, or nucleic acid. A functional equivalent of a protein may contain modifications depending on the necessity of such modifications for the performance of a specific function. The term “functional equivalent” is intended to include the “fragments,” “mutants,” “hybrids,” “variants,” “analogs,” or “chemical derivatives” of a molecule. “Compound electrified fluid jet” as used herein generally refers to a stream of fluid composed of at least two distinct fluid layers. For example, when two fluids are used to form a compound electrified jet by EHD, the outer fluid is the fluid precursor for the shell of the capsule made by the compound electrified fluid jet EHD method and the inner fluid is the precursor for the core of the capsule.
According to the invention, an agent is“encapsulated” when the capsule has at least one core region and at least one particle shell region in a layered architecture. The capsule may further contain at least one agent, such as a therapeutic agent or an imaging agent.
According to the invention, an agent is “entrapped” when the agent is dispersed within a biocompatible matrix composed of one or more biocompatible polymers, for example, and distinct onion-like layers are not apparent.
The term “carcinoma” as used herein generally refers to malignant tumors derived from epithelial cells. This group may represent the most common cancers, including the common forms of breast, prostate, lung and colon cancer.
The “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 tumors derived from the mesothelial cells lining the peritoneum and the pleura.
The term “glioma” as used herein generally refers to tumors derived from glia, the most common type of brain cell.
The term “germinoma” as used herein generally refers to tumors derived from germ cells, generally found in the testicle and ovary.
The term “choriocarcinoma” as used herein generally refers to malignant tumors derived from the placenta.
The term “attached,” as used herein generally refers to covalent binding, adsorption, and physical immobilization. The terms “associated with,” “binding” and “bound” are identical in meaning to the term “attached.”
The term “nanoparticle” as used herein generally refers to a particle, generally metallic, semiconducting, magnetic, ceramic and dielectric, having a diameter in the range of about 1 nm to about 1000 nm.
The terms “polymer” or “biopolymer”, as used herein generally refer to a compound having two or more monomer units, and is intended to include linear and branched polymers, and copolymers, the term “branched polymers” encompassing simple branched structures as well as hyperbranched and dendritic polymers. The term “monomer” is used herein to refer to compounds that are not polymeric. “Polymers” or “biopolymers” herein may be naturally occurring, chemically modified, or chemically synthesized.
The term “functional group” generally refers to compounds that may be suitable for chemically binding of a first and second molecule together. Chemical bonding may be considered to broadly cover bonding with some covalent character with or without polar bonding and can have properties of ligand-metal bonding along with various degrees of ionic bonding. The functional group may refer to ligand-receptor binding relationships were covalent bonding may not be the typical association. The functional group may be selected based on the composition of the molecule. Another functional group of the linker may be suitable for covalent bonding a first and second molecule together. Covalent bonding refers broadly to covalent bonds with sigma bonds, pi bonds, hydrogen bonds, other delocalized covalent bonds and/or other covalent bonding types, and may be polarized bonds with or without ionic bonding components and the like. Covalent linkers include functionalized organic molecules. The functional groups may include hydroxyl groups, amino groups, carboxyl groups, carboxylic acid anhydride groups, mercapto groups, and hydrosilicon groups.
The invention relates to systems and methods for fabrication of an agent, such as a therapeutic agent or an imaging agent, entrapped or encapsulated in a vehicle, capsule, particle, vector, or carrier in order to maximize treatment and/or imaging of malignancy while minimizing the adverse effects of treatment and/or imaging. Particularly, the systems and methods of the invention may form at least a two-liquid electrified jet, as described in
In one embodiment of the invention, the capsules may serve multiple purposes. First, the capsules may protect the encapsulated agent from biological attack and degradation before it reaches the target site. Second, the capsules may prevent surface interaction of the agent from inducing undesired signal pathways and introduce the generation of a cascade of proteins or hormones within the target cells thereby limiting the agents' effectiveness.
The capsule(s) having the entrapped or encapsulated agents may be used for the treatment or imaging of cancer in a patient. For example, the capsule may have an entrapped or encapsulated therapeutic agent for the treatment of malignant cancers such as lymphoma, leukemia, sarcoma, mesothelioma, glioma, germinoma, and choriocarcinoma. Furthermore, the capsule may have an entrapped or encapsulated imagining agent for the detection of cancer. The capsule of the invention may also have a combination of entrapped or encapsulated therapeutic agents and imaging agents.
There is no requirement that capsule 200 is spherical, other configurations and shapes, such as oblong, tubular, and ellipsoid may also be suitable for use in the invention. Additionally, capsule 200 may be configured as an independent particle or may be configured to be a series of capsules attached to each other in a chain-like fashion. In particular, if capsule 200 is configured in a chain-like series, the capsules may have a length in the range of about 0.03 μm to about 30 μm. Moreover, capsule 200 may be also be effective if it has an oblong shape, tubular shape or ellipsoid shape, if the length such as a minor axis in the case of an ellipsoid, or a diameter in the case of a tube, is sufficiently small to allow passage through the target site, such as a malignant cell membrane. The diameter or minor axis of the capsule may be in a range of about 10 nm to about 1 μm.
According to one embodiment of the invention, the architecture of the capsule may be based upon the mass ratio of shell material to core material and the composition and mass of the agents such as therapeutic agents, imaging agents, and other constituents, for example. Furthermore, the distinct core and shell regions may not be required. One or more agents, such as therapeutic or imaging agents may be entrapped and/or dispersed inside particles having at least on biocompatible substance such as a biopolymer. The capsule may include other additives, such as, a chemically functionalized surface to exploit receptor biochemistry properties of malignant cells, dispersed particles that may aid in the breaking or segregation of the therapeutic and/or an imaging agent carrying capsules once the capsules are substantially proximate to or inside the malignant cells.
According to an embodiment of the invention, the capsule may have a shell thickness in the range of about 10 nm to about 10 μm, and more particularly, in a range of about 20 nm to about 150 nm. In particular, the capsule shell has a thickness of about 20 nm. Moreover, the capsule may have a size in the range of about 25 nm to about 25 μm, and more particularly, in a range of about 50 nm to about 500 nm. Specifically, the capsule has a size of about 250 nm.
In another embodiment, the capsule including at least one agent, such as a therapeutic agent or an imaging agent, may be transported into the target cells by processes, such as endocytosis, transformation, or transduction. Once the capsule enters the target cell, the action of the therapeutic agent may induce the desired effect, such as apoptosis of the target cells. The therapeutic agents may be genetic material such as nucleic acid, RNA, DNA, bacteria, viruses, proteins, nucleic acid fragments, nucleic acid encoding a gene product, and abiological agents. Examples of the imaging agents may include magnetic particles, photonic sensitive materials, radioactive isotopes, and contrasting agents.
The capsule of the invention may have magnetic nanoparticles of low toxicity dispersed in it for external magnetic guidance of the capsule. In particular, the magnetic nanoparticles may include ferromagnetic materials, such as metallic iron and certain metal oxides, such a transition metal oxide, and rare earth based magnetic materials. Specifically, the magnetic nanoparticle may be a magnetite, such as Fe3O4 having a size in the range of about 1 nm to about 300 mn, and specifically, a size of about 10 nm. In addition to magnetic nanoparticles, other materials may be dispersed within the capsule such as materials sensitive to light or electric fields such as metallic silver or gold, which may be used for imaging effects. Moreover, the metallic silver or gold may be used in combination with radioactive emitters such as beta, gamma, or alpha emitters for imagining effects.
Referring to
The process throughput may be increased by making a device including more than one tube operating as tube 310, as described below. Depending on the physical and chemical properties of the shell fluid 308 and the core fluid delivered through tube 310, the height difference between the open end 322 of tube 310 and the surface of shell fluid 308 in the fluid bath 306, the reaction of core and shell fluids in the upwards traveling electrified liquid jet may be varied.
Moreover, in a further embodiment, a collector electrode 312 may be placed above the surface of shell fluid 308 in fluid bath 306 in a variety of orientations. For example, if a flat metal surface is used as collector electrode 312, the surface of the collector electrode may be oriented parallel or non-parallel to the surface of shell fluid 308 in fluid bath 306, or a different type of collector electrode configuration may be selected, such as a rotating drum.
The plume 328 may contain same-charge droplets that repel each other and thus, travel toward collector electrode 312 with an ever-increasing cross-sectional area. The electric potential of the core fluid and the electric potential of the shell fluid may be the same, due to their intimate electrical contact. The electrical potential and the voltage are related quantities. The electrical potential may depend upon the separation between the point at which a cone-jet region is formed, such as region 326, and electrode collection plate 312. The separation may have a distance in a range of about 1.5 inches to about 15 inches. Moreover, the applied voltage may be in a range of about 0.5 kV to about 35 kV.
Turning to
In an alternate embodiment, the direction that the electrified two-fluid ejected need not be upwards. Similar wetting by capillary forces of tubes or conduits delivering the core fluid may be effected to produce downward flows of the two-liquid electrified fluid jets. Electrified fluid or solid structures may be made to undergo total or partial electrical discharge at a region after the extractor. This configuration may increase the time of travel of the fluid or sold structures from the region where the electrified meniscus or menisci are formed, to the collection zone, because a smaller electrical charge means decreased mobility. The totally or partially discharged fluid or solid structures may be transported to a collection zone via use of gas flows.
The concept of wetting the external wall of a tube or conduit used to deliver the core fluid for a two-liquid electrified fluid jet may adopt other configurations. Referring to
For example,
In another embodiment, the concept of wetting the tube or conduit delivering the core fluid may be extended to other practical configurations.
Moreover, gap 718 and gap 720 may be adjusted or designed based upon operating variables such as, but not limited to, flow rates of the two fluids involved and the electric field strength and its spatial distribution. In particular, the closer electrified menisci are packed i.e., the smaller gap 720 is made, the higher the electric field required for forming the menisci. Unless the gas environment surrounding the electrified menisci is not air, corona discharges may occur, and these may compromise process continuity. Thus, a person of skill in the art would determine gap 720 based on these principles, on knowledge of the physical and chemical properties of the core and shell fluids, and the process variables. For example, if the physical and chemical properties of the core and shell fluids and the values of the operating variables require gap 720 to be long enough to cause dripping of the shell fluid in between adjacent tubes 710, flow deflectors or baffles may be incorporated between adjacent tubes 710 to convey the outer fluids towards tubes 10. Gap 718 may be determined by the wetting phenomena occurring between the outer walls of tubes 710 and the shell fluid 716. Additionally, an extractor electrode orifice may be placed at a short distance away from the corresponding two-fluid electrified meniscus, as described above, and may be used to facilitate formation of two-fluid electrified structures.
Referring to
Packing side-by-side a plurality of the fixture shown in
In one embodiment of the invention, the concept of wetting the outer wall of a tube or conduit delivering fluids by the action forced flow and electric fields can be extended to produce multi-fluid electrified menisci and jets. Referring
Moreover, a fixture comprising an arrangement of a plurality of tubes or conduits 1002 may be employed. The cross sectional areas of the tubes 1002 delivering the core liquid need not be circular, as other shapes such as, but not limited to, ellipsoidal and polygonal, may produce electrified two-fluid menisci and jets, and the open, cross sectional area of the tube need not be oriented perpendicular to the axis of the tube. The open end of the tube may adopt other shapes such as, but not limited to, conical or beveled. Tube 1002 may be substituted by a multi-tube arrangement consisting of more than one tube with cylindrical cross arranged in a coaxial manner, as described above, to deliver more than two fluids via forced flows and electric fields. The concept of placing an extractor electrode orifice at a short distance away from the corresponding electrified fluids meniscus, as described above, may be used expand the system illustrated in
Turning to
In one embodiment, once a two-fluid electrified liquid jet is formed by the methodology of the invention, a variety 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 such structures from a two-fluid electrified jet. For example, one way of making structures with defined core and shell regions of different composition, requires that the shell fluid undergo total or partial solvent evaporation during the time of travel from ejection of the two-fluid electrified jet from the two-fluid electrified meniscus to the collector electrode or collection zone. The precursor for the solid shell may remain in solution until a critical fraction of solvent evaporates to the surroundings. The core fluid, depending on how it is formulated by the skilled person, may or may not yield a solid phase on collection at the collection zone.
The methodology of the invention may be employed to produce structures with defined core and shell regions from a wide variety of fluids with different chemical compositions. Additional functions such as, but not limited to, magnetic properties may be added to the core or the shell fluids, or both, by formulating the chemistry of the said fluids.
For example, the shells of capsules containing a core fluid with dissolved protein may be made magnetic by suspending magnetite particles in the range of about 100 nm to about 1 nm in the shell fluid precursor prior to processing using the methods described herein. The core fluid and the shell fluid chemistry may b formulated in such a way that neither the core fluid nor the shell fluid undergoes solidification before entering the droplet-in-droplet regime depicted in
A variety of proteins, DNA material, cells and other biologically originated matter may be encapsulated using the methodology of the invention. In particular enzymes are a class of proteins that may serve as biocatalysts, and may be encapsulated in fluid cores such as, but not limited to, pH buffered aqueous solutions, and the shells may be formed from sol-gel precursors having suspended magnetite particles. The magnetite mass fraction in the shell may permit magnetic transport of the capsules once suspended in fluid media. The capsules containing the encapsulated enzymes may be suspended in a fluid containing reactants and products for a reaction catalyzed by the encapsulated enzyme. If the shell is designed with pores large enough to allow diffusion through it of reactants and products for the reaction catalyzed by the encapsulated enzyme, but not as large so as to permit the enzyme to diffuse out of the capsule, it is possible to recover the encapsulated enzyme with the aid of magnetic forces after the desired extent of reaction has been achieved.
For example, in one embodiment of the invention, silicon alkoxide sol-gel chemistry may produce pores in the capsules in the range of about 0.5 nm to about 2.0 nm when controlled, which may be sufficient to allow diffusion of many commercially important reactants and products, but not to allow, for example, about a 50 kDa to about a 60 kDa transaminase enzyme to escape out of the sol-gel derived shell. A natural extension of this concept is to design core fluid formulations containing substances of biological origin other than proteins such as, but not limited to DNA, DNA fragments, genes, and cells.
According to one embodiment, a magnetite nano-particle formulation compatible with the silicon alkoxides based shell chemistry may yield magnetite may be coated with a silicon alkoxide layer having a size in the range of about 2 nm to about 4 nm. Such a silicon alkoxide layer makes it ideal for firmly anchoring the magnetite to the shell solid phase via cross-linking sol-gel processes. A specific mass fraction of magnetite phase in the shell is in the range of about 0.1 to about 0.8.
In another embodiment of the invention, the capsule suitable for the treatment and/or imagining of malignant cancers may be fabricated by employing electrospray. In general, the electrospray methodology may include delivering a liquid containing the encapsulate(s) through a single nozzle, capillary, conduit, or orifice, applying an electrical potential between the nozzle and a collection region (herein after “counter-electrode”), forming a DC electrospray for the liquid containing the encapsulate(s), accelerating the DC electrospray towards the counter-electrode, forming the shell enveloping the DC electrospray droplets to produce vesicles. The vesicle may be formed during the time of flight of the DC electrospray from its delivery region to the counter-electrode through a reaction between a suitable shell-forming monomer with liquid components in the electrospray. The shell-forming monomer and the liquid components do not compromise the integrity and function of the encapsulate(s). The capsules may be collected on the counter-electrode are collected for storage and use.
Referring to
In one embodiment, the electrospray may be delivered into a region in space whose gaseous environment has an adequate concentration of shell-forming monomer. As shown in
In another embodiment of the invention, the encapsulate-containing electrospray may be passed through a solution containing the shell-forming monomer, instead of a gas containing the said monomer.
In another embodiment, the electrospray method of the invention may be made discontinuous, and the time of suspension of the electrically charged spray within the cyanoacrylate containing gaseous volume may be extended by applying a high voltage AC potential within the said shell-forming reaction volume, as shown in
In an alternative embodiment of the invention, a single fluid electrified jet may be employed, in the form of a solution or an emulsion to produce capsules having at least one entrapped agent. An emulsion, as used herein, generally refers to a single fluid that may be used to generate the particles instead of capsules with the EHD method as described above, using the same concentration ranges of the therapeutic components and other components, and the same average particle sizes as those of the capsules described below. Furthermore, magnetic nanoparticles may also be dispersed in the particles.
For example, an aqueous emulsified phase containing at least one agent, such as a therapeutic a/or imaging agent, and stabilizing salts and polyelectrolytes, may be dispersed in organic solutions containing at least one biopolymer. The resulting emulsion includes a biphasic medium which does not form distinct layers on electrified jet formation. The aqueous phase may be formulated in an identical manner as that of the core fluid in the case of the capsules described above, and the organic phase containing the biopolymer or biopolymers, which may also include magnetic nanoparticles and/or photonic particles formulated as that of the shell fluid of the capsules, as described above. Alternatively, a single fluid electrified jet formulation using certain co-solvents such as, but not limited to, dimethyl sulfoxide, leads to a homogeneous liquid mixture or solution comprising both the biopolymeric matrix precursor or precursors, and the active ingredients and receptors. The single electrified liquid jet approach may lead to entrapped, instead of encapsulated, active agents.
According to one embodiment, the shell-forming monomers may include compounds in the cyanoacrylate family, such as alkyl-substituted cyanoacrylates, for example. Moreover, the encapsulate-containing formulations may be compounds that readily react with the cyanoacrylate monomer to yield a skin of shell during the time-of-flight of the electrospray toward the counter-electrode, without chemically altering the intended function of the encapsulate. In particular, the encapsulates may contain water. The encapsulates may be of biological origin, such as nucleic acids. DNA, DNA fragments thereof, RNA, RNA fragments thereof, proteins, lipids, carbohydrates, and combinations thereof and/or any modification thereof. Additionally, the encapsulates may include abiological therapeutic drugs, as described below.
According to one embodiment of the invention, he biopolymers that may be suitable for producing the shell of the EHD-derived capsules include, without limitation, PLA, PLGA, chitosan, alkyl metacrylates, and other bioabsorbable polymers such as PCL, starch, and co-polymers resulting from any combination thereof. Moreover, PEG may also be incorporated, either as polymer chains or as co-polymeric fragments to one or more of the other biopolymers, or functionalized with groups such as thio, carboxylic acid and amine groups. Additionally, other constituents may be incorporated in the shell fluid, such as nanosized magnetite particles having a size in the range of about 1 nm to about 300 nm and surfactants, prior to electrospray to yield a magnetically sensitive MRI material. Furthermore, nanosized photonic sensitive silver, gold, palladium or a combination thereof, having a size in the range of about 1 nm to about 50 nm may be added to the shell precursor fluid prior to electrospray to yield a photonic sensitive material.
According to another embodiment, core fluids suitable for the methodology of the invention may include aqueous phase with buffer salts and polyelectrolytes such as phosphate buffers to stabilize the solution structure of therapeutic agents of biological origin, one or more therapeutic agents such as, but not limited to, tumor necrosis factor alpha (TNF-α) protein, or TNF-α, encoding cDNA 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 chemotherapy agents such as, but not limited to, acridinyl, anisidide, allopurinol, altretamine, aminoglutethimide, androgen, arsenic trioxide, asparaginease, azacitidine, antiangiogenesis, bleomycin, bortezomib, busulfan, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, cyclophasphamide, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, estramustine, etanidazole, etoposide, floxurdine, fludarabine, 5-fluorouracil, flrosol DA, Ftorafur, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, hydroxyurea, idarubicin, ifosfamide, interferon alpha, interleaukin-2, irinotecan, levamisole, lomustine, mechloroethamine, melphalan, menogaril, methotrexate, methyl-CCNU, mitomcycin, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, picamycin, procarbazine, raltitrexed, steptozocin, ftorafur, temozolomide, teniposide, thalidomide, thioguanine, thiotepa, topotecan, tositumoab, trimetrexate, valspodar, vinblastine, vincristine, vindesine, vinorelbine, combinations thereof, and functional equivalents thereof.
Moreover, the capsules or particles of the invention may contain chemotherapy-enhancing drugs, such as but not limited to, xcytrin, and chemotherapy protective drugs, such as but not limited to, amifostine. Certain radioactive elements may be suitable for the treatment and/or imaging of cancer, and in one embodiment of this invention, the radioactive elements can be incorporated in the electrospray generated particles. For example, Actinium-227 isotopic parent of Radium-223 is a so-called alpha emitter employed to treat metastases in the skeleton deriving from breast and prostate cancers, and for radioimmunotherapy, Copper-67 a gamma and beta emitter utilized in cancer radioimmunotherapy and lymphoma and colon and breast cancer diagnostics, Iodine-131, may be used for breast cancer treatment, leukemia, and lymphoma, using radioimmunotherapy, thyroid function and disease (e.g., cancer) studies, as well as non-malignant thyroid ailments (e.g. hyperthyroidism), Iodine-125, may be useful for implants for breast and prostate tumors, radiolabeling, prostate cancer brachytherapy, osteoporosis detection, tracer drugs, imaging diagnostics, mapping of brain receptors, brain cancer therapy, interstitial radiation treatment, determination plasma volume and glomerular filtration rate, deep vein thrombosis detection, Rhenium-186, useful for treatment and diagnosis of breast, colon, liver cancers and lymphoma via radioimmunotherapy, rheumatoid arthritis therapy, and bone cancer pain relief, and Yttrium-91, which is a gamma-emitting trace for Yttrium-90, which in turn is utilized for breast cancer radioimmunotherapy, and also lymphoma, kidney, colon, lung, prostate, ovarian, pancreatic, and liver cancers radioimmunotherapy.
In a further embodiment, radioactive isotopes may incorporated into the capsules of the invention, and their most frequent uses in medicine and related fields are: Cadmium-109 for general cancer detection, Actinium-225 and Thorium-229, the latter is the parent of Actinium-225 and grandparent of Bismuth-213 which are alpha emitters used in cancer radioimmunotherapy, Bismuth-212, and Thorium-228, the latter is the parent of Bismuth-212 which is an alpha emitter employed in cancer radioimmunotherapy, Cobalt-60, which is a radiation source for cancer radiotherapy, for food and medical supplies irradiation, Copper-64, a positron emitter employed for cancer therapy, Dysprosium-166, employed in cancer radioimmunotherapy, Erbium-169, useful for small-joint rheumatoid arthritis therapy, Europium-152 and Europium-154, useful as radiation sources for medical supplies and food irradiation, Gadolinium-153, useful for osteoporosis evaluation, Gold-198, useful for prostate and brain cancers, and for implants and intracavity treatment of ovarian cancer, Holmium-166, useful for myeloma therapy in targeted skeletal therapy, ablation of bone marrow, radioimmunotherapy of cancer, and rheumatoid arthritis therapy, Iridium-192, useful for spinal cord and brain tumor treatment, brachytherapy, and treatment of blocked blood vessels, Lutetium-177, useful for treatment of blocked blood vessels and cancer radioimmunotherapy, Molybdenum-99which is parent of Technetium-99m, which in turn is used for liver, brain, lungs, and heart imaging, and deep vein thrombosis detection, Osmium-194, useful for cancer radioimmunotherapy, Palladium-103, used in prostate cancer therapy, Platinum-195m, used in metabolism and biodistribution of cisplatin studies, Phosphorus-32, used for treatment of leukemia and polycythemia rubra vera (blood cell disease), bone cancer treatment and diagnosis, treatment of pancreatic, colon and liver cancer, treatment of blocked blood vessels and intracavity therapy, and diagnosis of surface tumors, Phosphorus-33, useful for leukemia therapy, treatment and diagnosis of bone disease, treatment of blocked blood vessels, and radiolabeling, Rhenium-188 useful for radioimmunotherapy, treatment of rheumatoid arthritis, bone cancer pain relief, and prostate cancer therapy, Rhodium-105, used for cancer radioimmunotherapy. Samarium-145, used for treatment of ocular cancer, Samarium-153, useful for cancer radioimmunotherapy, and bone cancer pain relief, Scandium-47, useful for bone cancer pain relief and cancer radioimmunotherapy, Selenium-75, useful for detection of hyperactive parathyroid glands, as radiotracer in brain evaluations, adrenal cortex imaging via gamma scintigraphy, detection of steroid producing tumors, and pancreatic scans, Strontium-85, useful for brain scanning and bone cancer detection, Strontium-89 useful for treatment of multiple myeloma and bone cancer pain relief, Thulium-170, used as energy source for medical device implants, and as a gamma source in blood irradiators, Tin-117m, used in bone cancer pain relief and cancer immunotherapy, Tungsten-188, which is the parent of Rhenium-188, which in turn is used for cancer treatment and diagnosis, rheumatoid arthritis treatment, and treatment of blocked blood vessels and bone cancer pain relief, Xenon-127, used in pulmonary function evaluation, brain disorder imaging, and brain blood flow analysis. Ytterbium-175, used in cancer radioimmunotherapy, and Yttrium-90, which is obtained from Yttrium-89 and is useful for liver cancer treatment. Metallic or nonmetallic nanoparticles such as Ga and B which are used in neutron activation treatment can be co-encapsulated with therapeutic agents. The capsules or particles of this invention may optionally contain chemotherapy-enhancing drugs, such as but not limited to, xcytrin, and chemotherapy protective drugs, such as but not limited to amifostine.
In another embodiment, the capsules of the invention may include at least one surface biochemical that may attach chemically or physically to enhance the affinity of the capsules for glioma cells, which may include epidermal growth factor, or EGF. The epidermal growth factor may either be incorporated onto the surface of the capsules post EHD processing, or during EHD processing as part of the shell fluid precursor. In particular, content range of EGF in the capsules and particles made by the methodologies of the invention may be in the range of about 0 to about 0.02 wt. %. Moreover, other surface biochemicals and chemicals may include, intraleukin-13, intraleukin-4, peripheral type benzodiazephine, vascular endothelial growth factor, platelet-derived growth factor, fibroblast growth factors, urokinase-type plasminogen activator, folic acid and folic acid derivatives, neurotrophin growth factor, somatostatin, cBSA, cHSA, protamine, insulin, ApoE derived peptide, polysorbate-80, OX-26, transferring, glucose, mannose, RMP-7, thiamine, combinations thereof and functional equivalents thereof. The surface biochemicals may be of human, animal or recombinant origin and have a content in the range of about 0 to about 1 wt % for the non-estrogen based surface biochemicals.
In a further embodiment, the capsules may include at least one surface biochemical that may attach chemically or physically to enhance the affinity of the capsule for lymphoma, myeloma and leukemia such as VEGR-2, tositumomab, a monoclonal antibody which binds to the CD20 receptor, and other antibodies and monoclonal antibodies in particular those designed for other clusters of differentiation, such as but not limited to, CDS, CD7, CD13, CD19, CD22, CD33, CD52, and CD61, and antimyeloperoxidase any combinations thereof, and functional equivalents thereof. These biochemicals and their derivatives may either be incorporated onto the surface of the capsules post EHD processing, or during EHD processing as part of the shell fluid precursor. The content range of the surface receptors in the capsules is in the range of about 0.0 to about 1.0wt %. Additional biochemicals and chemicals that may incorporate into the capsule may 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, epiregulin, and neuregulins, human epidermal growth factor 2 (HER-2) and family members, erbB2 and erbB1 receptor family, intraleukin-13 and family derivatives, platelet-derived growth factor, urokinase-type plasminon activator, folic acid and folic acid derivatives, neurotrophin growth factor, somatostatin, combinations thereof, and functional equivalents thereof. The surface biochemicals may be of human, animal or recombinant origin, and may be present in a content range of about 0.0 to about 1.0 wt %.
In a yet further embodiment, the capsules may include at least one surface biochemical that may attach chemically or physically to enhance the affinity of the capsule for breast cancer is estrogen. Estrogen and its chemical derivatives may either be incorporated onto the surface of the capsules post EHD processing, or during EHD processing as part of the shell fluid precursor. The content range of estrogen or its derivates in the capsules and particles may be in the range of about 0.02 to about 0.4 wt %. Other surface biochemicals and chemicals that may be incorporated into the surface of the capsules and particles may include human epidermal growth factor 2 (HER-2) and family members, erbB2 and erbB1 receptor family, intraleukin-13 and family derivatives, vascular endothelial growth factor, platelet-derived growth factor, fibroblast growth factors, urokinase-type plasminon activator folic acid and folic acid derivatives, neurotrophin growth factor, somatostatin, combination thereof, and functional equivalents thereof. The surface biochemicals may be of human, animal or recombinant origin, and may be in a content range of about 0 to about 1 wt %.
In another embodiment, the capsules may include surface biochemical that may attach chemically or physically to enhance the affinity of the capsule for pancreatic cancer cells may be 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, epiregulin, neuregulins, and functional equivalents thereof. These biochemicals and their derivatives may either be incorporated onto the surface of the capsules post EHD processing, or during EHD processing as part of the shell fluid precursor. The content range of these surface receptors in the capsules or particles may be in the range of about 0.0 to about 1.0 wt %. Additional surface biochemicals and chemicals that may be incorporated into the surface of the capsules and particles may include human epidermal growth factor 2 (HER-2) and family members, erbB2 and erbB1 receptor family, intraleukin-13 and family derivatives, platelet-derived growth factor, urokinase-type plasminon activator, folic acid and folic acid derivatives, neurotrophin growth factor, somatostatin, combinations thereof, and functional equivalents thereof. The surface biochemicals may be of human, animal or recombinant origin, and may be in a content range of about 0 to about 1 wt %.
Moreover, other surface biochemicals and chemicals for the said capsules and particles may be fused in signal sequences, mitochondrial, nuclear, actin, and tubulin, golgi, plasma membrane, peroxisome, and may be in a 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 capsules or particles may be in the range of about 10 to about 3200 μg/mL. Specifically, a concentration of TNF-α protein in the capsules or particles may be in the range of about 100 μg/mL to about 1,000 μg/mL. More specifically, a concentration of Egr-TNF in the capsules or particles may be in the range of about 10 μg/mL to about 300 μg/mL. The therapeutic effects may also be achieved with capsules containing either one or two compounds of the three-component, Paclitaxel, TNF-α and Egr-TNF group. Narrow ranges of the mass fractions of the therapeutic components and other capsules components, the capsules average size, average shell thickness, and narrow core to shell average mass ratios may be selected based on the location and size of the malignancy, and use magnetic nanoparticle doping to aid in the guiding of the said capsules to the malignancy may not be desired. For example, direct tumor injection, may be used in place of magnetically assisted delivery, and electromagnetic irradiation of the malignancy in contact with the particles or capsules by the methodologies of the invention may be used to trigger disruption of the particle with a concomitant release of therapeutic agents, to initiate biochemical processes such as, but not limited to, malignant cell DNA damage, or a combination of the said electromagnetic irradiation induced processes. DNA markers containing pCMV-Luc+plasmid containing the cytomegalovirus (CMV) promoter of pcDNA3 inserted upstream to the firefly luciferase of the pGL2basic vector plasmid and TK renilla luciferase may also be used as markers.
In an alternate embodiment, convection enhanced delivery, or CED, may be used in place of magnetically assisted delivery, and electromagnetic irradiation of the malignancy in contact with the particles or capsules made by the methodologies of the invention may be used to trigger disruption of the capsule or particle with a concomitant release of therapeutic agents, to initiate biochemical processes such as, but not limited to, malignant cell DNA damage, or a combination of the said electromagnetic irradiation induced processes.
The agents suitable for encapsulation or entrapment in the capsules of the invention should not be construed to be limited to agents suitable for cancer treatment or imaging of malignancy, as described above. However, alternative agents may be suitable for entrapment or encapsulation may include anti-inflammatory compounds, anti-allergics, glucocorticoids, anti-infective agents, antibiotics, antifungals, antivirals, mucolytics, antiseptics, vasoconstrictors, wound healing agents, local anaesthetics, 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, desoxymethasone, fluconolone acetonide, flucinonide, flunisolide, fluticasone, icomethasone, rofleponide, triamcinoline acetonide, fluocortin, butyl, hydrocortisone, hydroxycortisone-17-butyrate, prednicarbate, 6-methylprednisolone aceponate, mometasone furoate, elastane-, prostaglandin-, leukotriene, bradykinin-antagonists, non-steroidal anti-inflammatory drugs (NSIDs), such as ibuprofen, indometacin, including any pharmaceutically acceptable salts, esters, isomers, steroisomers, diastereomers, epimers, solvates or other hydrates, prodrugs, derivtives, or any other chemical or physical forms of active compounds comprising the respective active moieties.
Examples of potentially useful antiallergic agents include the afore-mentioned glucocorticoids, and nedocromil, cetrizin, loratidin, montelukast, roflumilast, ziluton, omalizumab,heparins and heparinoids and other antihistamins, Azelastin, Cetirizin, Desloratadin, Ebastin, Fexofenadin, Levocetirizin, Loratadin.
Examples of antinfective agents, whose class or therapeutic category is herein understood as comprising compounds which are effective against bacterial, fungal, and viral infections, i.e. encompassing the classes of antimicrobials, antibiotics, antifungals, antiseptics, and antivirals, are penicillins, including benzylpenicillins (pencillin-G-sodium, clemizone penicillin, benzathine penicillin G), phenoxypenicillins (penicillin V, propicillin), aminobenzylpencillins (ampicillin, amoxycillin, bacampicillin), acylaminopenicillins (azlocillin, mezlocillin, piperacillin, apalcillin), carboxypenicillins (carbenicillin, ticarcillin, temocillin), isoxazolyl pencillins (oxacillin, cloxacillin, dicloxacillin, flucloxacillin), and amiidine penicillins (mecillinam); cephalosporins, including cefazolins (cefazolin, cefazedone); cefuroximes (cerufoxim, cefamdole, cefotiam), cefoxitins (cefoxitin, cefotetan, latamoxef, flomoxef), cefotaximes (cefotaxime, ceftiaxone, ceftizoxime, cefmenoxime), ceftazidimes (ceftazidime, cefpirome, cefeepime), cefalexins (cefalexin, cefaclor, cefadroxil, cefradine, loracarbef, cefprozil), and cefiximes (cefixime, cefpooxim proxetile, cefuroxime axetil, cefetarnet pivoxil, cefotiam hexetil), loracarbef, cefepim, clavulanic acid/amoxicillin, Ceftobiprole;synergists, including beta-lactamase inhibitors, such as clavulanic acid, sulbactam, and tazobactam; carbapenems, including impenem, cilastin, meropenem, coripenem, tebipenem, ertapenem, ritipenam, and biapentem; monobactams, including aztreonam; aminoglycosides, such as apramycin, gentamicin, amikacin, isepamicin, arbekacin, tobramycin, netilmicin, spectinomycin, streptomycin, capreomycin, neomycin, paromoycin, and kanamycin; macrolides, including erythromycin, clarythromycin, roxithromycin, azithromycin, dithromycin, josamycin, including erythromycin, clarythromycin, roxithromycin, azithromycin, dithromycin, josamycin, spiramycin and telithromycin; gyrase inhibitors or fluroquinolones, including ciprofloxacin, gatifloxacin, norloxacin, ofloxacin, levofloxacin, perflexacin, lomefloxacin, fleroxacin, garenoxacin, clinafloxacin, sitafloxacin, prulifloxacind, olamulfloxacin, caderofloxacin, gemifloxacin, balofloxacin, trovalfloxacin, and moxifloxacin; tetracyclins, including tetracyclin, oxytetracyclin,rolitetracyclin, minocyclin, doxycycline, tigecycline and aminocycline; glycopeptides, including vancomycin, teicoplanin, ristocetin, avoparcin, oritavancin, ramoplanin, and peptide 4; polypeptides, including plectasin, dalbavancin, daptomycin, oritavancin, ramoplanin, dalbavancin, telavancin, bacitracin, tyrothricin, neomycin, kanamycin, mupirocin, paromomycin, polymyxin B and colistin; sulfonamides, including sulfadiazine, sulfamethoxazole, sulfalene, co-trimoxazole, co-trimetrol, co-trimoxazine, and co-tetraxazine; azoles, including clotrimazole, oxiconazole, miconazole, ketoconazole, itraconazole, fluconazole, metronidazole, tinidazole, bifonazol, ravuconazol, posaconazol, voriconazole, and omidazole and other antifungals including flucytosin, griseofluvin, tonoftal, naftifin, terbinafin, amorolfin, cielopiroxolantin, echinocandins, such as micafungin, caspofungin, anidulafungin; nitrofurans, including nitrofurantoin and nitrofuranzone; polyenes, including amphotericin B, natamycin, nystatin, flucocytosine; other antibiotics, including tithromycin, lincomycin, clindamycin, oxazolindiones (linzezolids), ranbezolid, streptogramine A+B, pristinamycin aA+B, Virginiamycin A+B, dalfopristin/qiunupristin (Synercid), chloramphenicol, ethambutol, pyrazinamid, terizidon, dapson, prothionamid, fosfomycin, fucidinic acid, rifampicin, isoiazid, cycloserine, terizidone, ansamycin, lysostaphin, iclaprim, mirocin B17, clerocidin, filgrastim, and pentamidine; antivirals, including aciclovir, ganciclovir, birivudin, valaciclovir, zidovudine, didanosin, thiacytidin, stavudia, lamivudin, zalcitabin, ribavirin, nevirapirin, delaviridin, trifluridin, ritonavir, saquinavir, indinavir, foxcarnet, amantadin, podophyllotoxin, vidarabine, tromantadine, and proteinase inhibitors; plant extracts or ingredients, such as plant extracts from chamomile, hamamelis, ehinacea, calendula, papain, pelargoniium, essential oils, mytol, pinen, limonen, cineole, thymol, mentol, tee tree oil, alpha-hederin, bisabolol, lycopodin, vitapherole; wound healing compounds including dexpantenol, allantoin, vitamins, hyaluronic acid, alpha-antitrypsin, anorganic and organic zinc salts/compounds, interferones (alpha, beta, gamma), tumor necrosis factors, cytokines, interleukins.
Examples of potentially useful mucolytics are DNase, P2Y2-agonists (denufosol), heparinoids, guaifenesin, acetylecysteine, carbocysteine, ambroxol, bromhexine, lecithins, myrtol, and recombinant surfactant proteins.
Examples of potentially useful local anaesthetic agents include benzocaine, tetracaine, procaine, lidocaine and bupivacaine.
Examples of potentially useful antiallergic agents include the afore-mentioned glucocorticoids, nedocromil. Examples of potentially useful peptides and proteins include antibodies against toxins produced by microorganisms, antimicrobial peptides such as cecropinis, defensins, thionins, and cathelicidins.
Also, immunmodulators including methotrexate, azathioprine, cyclosporine A, tacrolimus, sirolimus, rapamycin, mycofenolate, mofetil, cytotatics and metastasis inhibitors, alkylants, such as nimustine, melphanlane, carmustine, lomustine, cyclophosphosphamide, ifosfamide, trofosfamide, chlorambucil, busulfane, treosulfane, prednimustine, thiotepa; antimetabolites, e.g. cytarabine, fluorouracil, methotrexate, mercaptopurine, tioguanine; alkaloids, such as vinblastine, vincristine, vindesine; antibiotics, such as alcarubicine, bleomycine, dactinomycine, daunorubicine, doxorubicine, epirubicine, idarubicine, mitomycine, plicamycine; complexes of secondary group elements (e.g. Ti, Zr, V, Nb, Ta, Mo, W, Pt) such as carboplatinum, cis-platinum and metallocene compounds such as titanocendichloride; amsacrine, dacarbazine, estramustine, etoposide, beraprost, hydroxycarbamide, mitoxanthrone, procarbazine, temiposide; paclitaxel, iressa, zactima, poly-ADP-ribose-polymerase (PRAP) enzyme inhibitors, banoxantrone, gemcitabine, pemetrexed, bevacizumab, ranibizumab may be suitable for entrapment or encapsulation in the capsules of the invention.
In a further embodiment other compounds may include proteinase inhibitors, such as a-anti-trypin; antioxidants, such as tocopherols, glutathion; pituitary hormones, hypothalamic hormones, regulatory peptides and their inhibiting agents, corticotropine, tetracosactide, choriogonandotropine, urofolitropine, urogonadotropine, saomatotrpine, metergoline, desmopressine, oxytocine, argipressine, omipressine, leuproreline, triptoreline, gonadoreline, busereline, nafareline, goselerine, somatostatine; parathyroide gland hormones, calcium metabolism regulators, dihydrotachysterole, calcitonine, clodronic acid, etidronic acid; thyroid gland therapeutics; sex hormones and their inhibiting agents, anabolics, androgens, estrogens, gestagenes, antiestrogenes; anti-migraine drugs, such as proxibarbal, lisuride, methysergide, dihydroergotamine, ergotamine, clonidine, pizotifene; hypnotics, sedatives, benzodiazepines, barbiturates, cyclopyrrolones, imidazopyridines, antiepileptics, zolpidem, barbiturates, phenytoin, primidone, mesuximide, ethosuximide, sultiam, carbamazepin, valpric acid, vigabatrine; antiparkinson drugs, such as levodopa, carbidopa, benserazide, selegiline, bromocriptine, amantadine, tiapride; antiemetics, such as thiethylperazine, bromopride, domperidone, granisetrone, ondasetrone, tropisetrone, pyridoxine; analgesics, such as buprenorphine, fentanyl, morphine, codeine, hydromorphone, methadone, fenpipramide, fentanyl, piritramide, pentazocine, buprenorphine, nalbuphine, tilidine; drugs for narcosis, such as N-methylated barbiturates, thiobarbituraates, ketamine, etomidate, propofol, benzodiazepines, droperidol, haloperidol, alfentanyl, sulfentanyl; antirheumatism drugs including tumor necrosis factor-alfa, nonsteroidal antiinflammatory drugs; antidiabetic drugs, such as insulin, sulfonylurea derivatives, biguanids, glitizols, glucagon, diazoxid; cytokines, such as interleukines, interferones, tumor necrosis factor (TNF), colony stimulating factors(GM-CSF, G-CSF, M-CSF); proteins, e.g. epoetine, and peptides, e.g. parathyrin, somatomedin C; heparine, heparinoids, urokinases, streptokinases, ATP-ase, prostacycline, sexual stimulants, or genetic material.
The description and examples given above are merely illustrative and are not meant to be 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 invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection 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 material sciences, polymer sciences, or related fields are intended to be within the scope of the appended claims.
The disclosures of all references and publications cited above are expressly incorporated by reference in their entireties to the same extent as if each were incorporated by reference individually.
EXAMPLES Specific Example 1This example described an encapsulation of a protein solution using a modification of the shell formulation that consisted of a solution of bovine serum albumin (BSA) in the presence of salts such as phosphates to stabilize the pH of the solution. Confocal fluorescence microscopy (CFM) and BSA with a fluorescent tag was used for visualizing the capsule. About 2 mg fluorescent protein was dissolved in about 1 mL of phosphate-buffer saline (PBS), type buffer that stabilized the acid-base properties of this aqueous solution to a pH equal to about 7.4.
The final BSA concentration was adjusted to about 3 μM. Silica sol was used as the shell fluid precursor following the sol aging procedures, with addition of tert-amyl alcohol to a 50:50 volume ratio.
In general, for solution aging purposes, an acidified tetraethyl orthosilicate solution in ethanol may be aged at about 80° C. for about 4 to about 6 hours. Ter-amyl alcohol was added to increase the hydrophobicity of the shell fluid, which further prevented any significant mixing between core and shell fluids.
The core and shell flow rates were adjusted to about 0.025 and about 0.75 mL/h, respectively. Adequate voltages for capsule design were in the range of about 11 to about 12 kV, and the distance between the two-fluid electrified meniscus and the collection zone was in the range of about 4 to about 14 cm. The collection zone, or collector electrode, was a flat metal surface kept at ground electrical potential, whereas the two-fluid electrified meniscus were kept at a positive electrical bias in the range of about 5 to about 18 kV.
The sol-gel methods and process variables used to encapsulate fluorescent BSA in Specific Example 1 were slightly modified to encapsulate a transaminase. This enzyme was used to catalyze the following reaction:
D.L glutamine+glyoxylic acid→L-glutamine (left unreacted)+α-keto derivative (from D-glutamine)-glycine
Each enantiomeric from has the ability to rotate polarized light, and a technique known as polarimetry may be used to follow chemical reactions in involving enantiomers as a function of time. The reactants, on the left hand side of the reaction shown above, are basically a mixture with no optical activity, since the D,L prefix stands for about a 50:50 mixture of the D and L enantiomers of glutamine. The products, on the right hand side of the reaction shown above, become enriched in unreacted L-glutamine with a concomitant time-dependent optical rotation signal that may be tracked by polarimetry, because this particular transaminase only catalyzes reactions involving D enantiomers. The reaction was buffered to a about pH of 7.5 with PBS.
Since the specific rotation of the glutamine isomers was low in pure water, an experimental scheme was devised to both stop the reaction prior to polarimetry quantification at different reaction times, and to increase the sensitivity of the analytical technique. This was achieved by addition of about 1.0 mL or 5 M HCl to an aliquot taken from the reactor, which denatures the enzyme. The enzyme catalyst either settles at the bottom of the vial, or was removed by centrifugation, and acidification increased the sensitivity of the polarimetry technique by about ½ order of magnitude relative to that observed in nearly neutral solutions.
The biocatalytic test was run using about 50 mM concentrations of the substrates, and enzyme concentrations of about 0.5 mg/mL.
The therapeutic solution was prepared by mixing 3.1 LACZ in about 10 mM Bis-Tris propane buffer aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of 3.1 LACZ was 700 μg/mL. A biopolymer solution was prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. This solution was doped with a solution of magnetite particles with an average diameter of about 15 nm. The weight percent contents of PEG-b-PLA, PCSH, and magnetite particles in this solution were about 0.071 wt %, about 0.058 wt %, and about 0.004 wt % respectively.
The therapeutic and biopolymer solutions were mixed and dimethyl sulfoxide, or DMSO, was added to form a homogenous solution. Particles with an average diameter in the range of about 0.250 to about 1 μm were made produced, but smaller capsules may be made by varying the process variables. Specifically, 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 2 NUC The compound in this example was formed by the two-jet system depicted in
The shell fluid solution was prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH in the shell fluid solution were about 0.29 wt % and about 0.31 wt %, respectively.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm were produced. Specifically, the core and shell fluid flow rates used were about 0.050 and about 0.300 mL/h, respectively, and the external voltage was about 7 kV.
Capsules with a PDs red 2 NUC loading in the range of about 0.01 to about 25% by weight of the added polymer may be made by this method by adjusting the concentration of the core fluid solution.
Specific Example 5 Encapsulation of Green Fluorescent ProteinThe core fluid solution was prepared by mixing Green Fluorescent Protein, or GFP, and about 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of GFP in the core fluid solution was about 30 μg/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, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH were about 0.32 wt % and about 0.31 wt %, respectively.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm were produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were about 0.050 and about 0.300 ml/h, respectively, and the external voltage was about 8 kV.
Capsules with a GFP loading in the range of about 0.01 to about 20% by weight of the added polymer were made by this method by adjusting the concentration of the core fluid solution.
Specific Example 6 Particles with Doxorubicin with Folic Acid Functional GroupsThe therapeutic solution is prepared by dissolving Doxorubicin hydrochloride, or DOXO, in dichloromethane. The concentration of DOXO in the therapeutic solution is about 1,000 μg/mL.
A biopolymer solution is prepared by mixing two functionalized biopolymers in chloroform: (a) Folate-functionalized poly(ethylene glycol), or Folate-PEG, with a molecular weight of 5,000 Da, and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of Folate-PEG and PCSH are about 0.30 wt % and about 30 wt %, respectively.
The therapeutic and biopolymer solutions are mixed to form a homogenous solution. Particles with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flowrates used are in the range of about 0.050 to about 0.300 ml/h. The external voltage is about 7.5 kV.
Particles with a DOXO loading in the range of about 0.01 to about 25% by weight of the added polymer may be made by this method by adjusting the concentration of the core fluid solution.
Specific Example 7 Encapsulation of DoxorubicinThe core fluid solution were prepared by mixing doxorubicin hydrochloride, or DOXO in about 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of DOXO in the core fluid solution was about 1,000 μg/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, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and(b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH were about 0.3 wt % and about 0.3 wt %, respectively.
Capsules with an average diamter in the range of about 0.250 μm to about 1 μm were produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were 0.050 and 0.300 ml/h, respectively, and the external voltage was 8.5 kV.
Capsules with a DOXO loading in the range of about 0.01 to about 23% by weight of the added polymer may be made by this method by adjusting the concentration of the core fluid solution.
Specific Example 8 Encapsulation of Cobalt NanoparticlesThe core fluid solution was prepared by mixing an aqueous solution of cobalt nanoparticles, or NP-Co and a 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of NP-Co in the core fluid solution was in the range of about 1,000 to about 500 μg/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, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH were about 0.30 wt % and about 0.30 wt %, respectively.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm were produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were in the range of about 0.050 to about 0.150 ml/h, respectively, and the external voltage was about 8 kV.
Capsules with a nanoparticle (Np)-Co loading in the range of about 0.01 to about 11% by weight of the added polymer were made by this method by adjusting the concentration of the core fluid solution.
Specific Example 9 Particles with Doxorubicin with EGF Functional GroupsThe therapeutic solution is prepared by dissolving Doxorubicin hydrochloride, or DOXO, in dichloromethane. The concentration of DOXO in the therapeutic solution is about 1,000 μg/mL.
A biopolymer solution is prepared by mixing two functionalized biopolymers in chloroform: (a) EGF-functionalized poly(ethylene glycol), or EGF-PEG, with a molecular weight of 5,000 Da, and (b) Poly(caprolactone)SH, or PCSH, with a molecular weight of 5,000 DA and a Mw/Mn=1.5. The weight percent contents of EGF-PEG and PCSH are about 0.30 wt % and about 0.30 wt %, respectively.
The therapeutic and biopolymer solutions are mixed to form a homogenous solution. Particles with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flowrates used are in the range of about 0.050 to about 0.300 ml/h range. The external voltage is about 7.5 kV.
Particles with a DOXO loading in the range of about 0.01 to about 25% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 10 Particles with Luciferase as DNA MarkerThe therapeutic solution is prepared by mixing Luciferase in a 10 mM Bis-Tris propane buffer aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of Luciferase is about 334.5 μg/mL.
A biopolymer solution is prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. This solution is doped with a solution of magnetite particles with an average diameter of about 15 nm. The weight percent contents of PEG-b-PLA, PCSH, and magnetite particles in this solution are about 0.072 wt %, about 0.058 wt %, and about 0.004 wt % respectively.
The therapeutic and biopolymer solutions are mixed and dimethyl sulfoxide, or DMSO, is added to form a homogenous solution. Particles with an average diameter in the range of about 0.250 to about 1 μm are produced, but smaller capsules may be made by varying the process variables. Specifically, the flow rate used was about 0.150 mL/h, and the external voltage was about 7 kV.
Particles with a Luciferase loading in the range of about 0.01 to about 63% by weight of the added polymer may be made by adjusting the concentration of the Luciferase-containing solution.
Specific Example 11 Particles with TK Renilla Luciferase as DNA MarkerThe therapeutic solution was prepared by mixing TK Renilla Luciferase in about 10 mM Bis-Tris propane buffer aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of TK Renilla Luciferase was about 334.5 μg/mL.
A biopolymer solution is prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. This solution is doped with a solution of magnetite particles with an average diameter of about 15 nm. The weight percent contents of PEG-b-PLA, PCSH, and magnetite particles in this solution are about 0.071 wt %, about 0.058 wt %, and about 0.004 wt % respectively.
The therapeutic and biopolymer solutions were mixed and DMSO was added to form a homogenous solution. Particles with an average diameter in the range of about 0.250 to about 1 μm were produced, but smaller capsules may be made by varying the process variables. Specifically, the flow rate used was about 0.150 mL/h, and the external voltage was about 7 kV.
Particles with a TK Renilla Luciferase loading in the range of about 0.01 to about 63% by weight of the added polymer may be made by adjusting the concentration of the TK Renilla Luciferase-containing solution.
Specific Example 12 Encapsulation of Cobalt Nanoparticles in Capsules Containing EGF and Mitochondria Localization VectorThe core fluid solution is prepared by mixing an aqueous solution of cobalt nanoparticles, or NP-Co and a 10 mM Bis-Tris propane aqueous solution containing about 1.0 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of NP-Co in the core fluid solution is in the range of about 1,000 to about 500 μg/mL.
The shell fluid solution is prepared by mixing three functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5; and (c) EGF-Functionalized poly(ethylene glycol), or EGF-PEG, with a molecular weight of 5,000 Da. The weight percent contents of PEG-b-PLA and PCSH and EGF-PEG are about 0.30 wt % and about 0.30 wt % and about 0.10 wt %, respectively.
A buffer solution containing a plasmid subcellular localization vector targeted to the mitochondria is added. The concentration of the vector in the shell fluid solution is in the range of about 0.0 to about 0.10 wt %, respectively.
A buffer solution containing a plasmid subcellular localization vector targeted to the mitochondria is added. The concentration of the vector in the shell fluid solution is in the range of about 0.0 to about 1.0 wt %. DMSO is added to form a homogenous solution.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were about 0.050 and about 0.150 ml/h, respectively, and the external voltage was about 8 kV.
Capsules with a Np-Co loading in the range of about 0.01 to about 11% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 13 Encapsulation of Chlorambucil in Capsules Containing CD19 Functional Groups on the ShellThe core fluid solution was prepared by dissolving Chlorambucil in about 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol, about 2 mM of CaCl2 DMSO. The final concentration of Chlorambucil in the core fluid solution was in the range of about 2,000 to about 500 μg/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, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH were about 0.05 wt % and about 0.05 wt %, respectively. The shell fluid solution was mixed with a solution containing CD19 dissolved in a mixture of dichloromethane and polyethylene oxide (MW=400-1000).
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm were produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were about 0.050 and about 0.150 ml/h, respectively, and the external voltage was about 8 kV.
Capsules with a Chlorambucil loading in the range of about 0.01 to about 20.0% by weight of the added polymer may be made by adjusting the concentration of the shell fluid solution.
Specific Example 14 Encapsulation of Chlorambucil in Capsules Containing CD20 Functional GroupsThe core fluid solution was prepared by dissolving Chlorambucil in a about 10 mM Bis-Tris propane aqueous solution containing about 1.0 wt % of isopropanol, about 2.0 mM of CaCl2 and DMSO. The final concentration of Chlorambucil in the core fluid solution was in the range of about 2,000 to about 500 μg/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, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH were about 0.05 wt % and about 0.05 wt %, respectively. The shell fluid solution was mixed with a solution containing CD20 dissolved in a mixture of dichloromethane and polyethylene oxide (MW=400-1000).
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm were generated, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were about 0.050 and about 0.150 ml/h, respectively, and the external voltage was about 8 kV.
Capsules with a Chlorambucil loading in the range of about 0.01 to about 20.0% by weight of the added polymer may be made by adjusting the concentration of the shell fluid solution. Capsules with a CD20 loading in the range of about 5 to about 100 μg per mg of the added polymer may be made by adjusting the concentration of the shell fluid solution.
Specific Example 15 Encapsulation of Chlorambucil and Hydrochloroquine Sulfate in Capsules Containing CD19 and CD20 Functional GroupsThe core fluid solution was prepared by dissolving Chlorambucil and Hydrochloroquine sulfate, of HCQ, in about 10 mM Bis-Tris propane aqueous solution containing about 1.0 wt % of isopropanol, about 2.0 mM of CaCl2 and DMSO. The final concentration of Chlorambucil and HCQ in the core fluid solution were in the range of about 2,000 to about 500 μg/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, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH were about 0.05 wt % and about 0.05 wt %, respectively. The shell fluid solution was mixed with a solution containing CD19 and CD20 dissolved in a mixture of dichloromethane and polyethylene oxide (MW=400-1000).
Capsules with an average diameter in the range of about 0.250 μm to about 1 were produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were about 0.050 and about 0.300 ml/h, respectively, and the external voltage was about 8 kV.
Capsules with a Chlorambucil and HCQ loading in the range of about 0.01 to about 20.0% each by weight of the added polymer may be made by adjusting the concentration of the core fluid solution. Capsules with a CD19 and CD20 loading in the range of about 5 to about 100 μg each per mg of the added polymer may by adjusting the concentration of the shell fluid solution.
Specific Example 16 Encapsulation of Chlorambucil in Capsules Containing CD19 Functional Group and Colgi Complex Localization VectorThe core fluid solution is prepared by dissolving Chlorambucil in about 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol, about 2 mM of CaCl2 and DMSO. The final concentration of Chlorambucil in the core fluid solution is in the range of about 2,000 to about 500 μg/mL.
The shell fluid solution is prepared by mixing two functionalized biopolymers in chloroform:(a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and(b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH are about 0.05 wt % and about 0.05 wt %, respectively. The shell fluid solution is mixed with a solution containing CD19 dissolved in a mixture of dichloromethane and polyethylene oxide (MW=400-1000).
A buffer solution containing a plasmid subcellular localization vector targeted to the Golgi complex is added. The concentration of the vector in the shell fluid solution is in the range of about 0 to about 1 wt %. DMSO is added to form a homogenous solution.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used are about 0.050 and about 0.150 ml/h, respectively, and the external voltage is about 8 kV.
Capsules with a Chlorambucil loading in the range of about 0.01 to about 20.0% by weight of the added polymer may be produced by adjusting the concentration of the core fluid solution. Capsules with a CD19 loading in the range of about 5 to about 100 μg per mg of the added polymer may be made by adjusting the concentration of the shell fluid solution.
Specific Example 17 Particles Containing Iodine-125 and EGF Functional GroupsThe therapeutic solution is prepared by dissolving Iodine-125 in dichloromethane. The concentration of Iodine-125 in the therapeutic solution is about 1,000 μg/mL.
A biopolymer solution is prepared y mixing two functionalized biopolymers in chloroform: (a) EGF-functionalized poly(ethylene glycol), or EGF-PEG, with a molecular weight of 5,000 Da, and (B) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of EGF-PEG and PCSH are about 0.30 wt % and about 0.30 wt %, respectively.
The therapeutic and biopolymer solutions are mixed to form a homogenous solution. Particles with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flowrates used are in the range of about 0.050 to about 0.300 ml/h. The external voltage is about 7.5 kV.
Particles with an Iodine-125 loading in the range of about 0.01 and 25% by weight of the added polymer may 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 ReceptorThe core fluid solution is prepared by dissolving Chlorambucil in a 10 mM Bis-Tris propane aqueous solution containing about 1.0 wt % of isopropanol, about 2.0 mM of CaCl2 and DMSO. The final concentration of Chlorambucil in the core fluid solution is in the range of about 2,000 to about 500 μg/mL.
The shell fluid solution is prepared by mixing two functionalized biopolymers in chloroform: (a) Epidermal growth factor-functionalized poly(ethylene glycol), or EGF-PEG, with a molecular weight of 5,000 Da, and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. This solution is doped with a solution of Fe3O4 and Ga2O3 nanoparticles with an average diameter of about 15 nm. The weight percent contents of EGF-PEG, PCSH, Fe3O4 and Ga2O3 in the shell fluid solution are about 0.071 wt %, about 0.058 wt %, about 0.004 wt % and about 0.004 wt %, respectively.
Particles with an average diamter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flow rates used are in the range of about 0.050 to about 0.300 ml/h. The external voltage is about 7.5 kV.
Particles with a Chlorambucil loading in the range of about 0.01 to about 25.0% by weight of the added polymer may be made 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 ReceptorThe core fluid solution is prepared by dissolving Chlorambucil in about 10.0 mM Bis-Tris propane aqueous solution containing about 1.0 wt % of isopropanol, about 2.0 mM of CaCl2 and DMSO. The final concentration of Chlorambucil in the core fluid solution is in the range of about 2,000 to about 500 μg/mL.
The shell fluid solution is prepared by mixing two functionalized biopolymers in chloroform: (a) Epidermal growth factor-functionalized poly(ethylene glycol), or EGF-PEG, with a molecular weight of 5,000 Da, and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. This solution is doped with a solution of Fe3O4, Ga2O3 and B2O3 nanoparticles with an average diameter of about 15 mn. The weight percent contents of EGF-PEG, PCSH, Fe3O4, Ga2O3 and B2O3 in the shell fluid solution are about 0.071 wt %, about 0.058 wt %, about 0.004 wt %, about 0.004 wt % and about 0.004 wt %, respectively.
Particles with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flowrates used are in the range of about 0.050 to about 0.300 ml/h. The external voltage is about 7.5 kV.
Particles with a Chlorambucil loading in the range of about 0.01 to about 25% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 20 Particles with Paclitaxel with Folic Acid Functional GroupsThe therapeutic solution is prepared by dissolving Paclitaxel in dichloromethane. The concentration of Paclitaxel in the therapeutic solution is about 1,000 μg/mL.
A biopolymer solution is prepared by mixing two functionalized biopolymers in chloroform: (a) Folate-functionalized poly(ethylene glycol), or Folate-PEG, with a molecular weight of 5,000 Da, and (B) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of Folate-PEG and PCSH are about 0.30 wt % and about 0.30 wt %, respectively.
The therapeutic and biopolymer solutions are mixed to form a homogenous solution. Particles with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flowrates used are in the range of about 0.050 to about 0.300 ml/h. The external voltage is about 7.5 kV.
Particles with a Paclitaxel loading is in the range of about 0.01 and 25% by weight of the added polymer may by adjusting the concentration of the core fluid solution.
Specific Example 21 Capsules with Paclitaxel with Folic Acid Functional GroupsThe core fluid solution is prepared by mixing Paclitaxel in DMSO and about 10 wt % of γ-cyclodextrine in about 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of Paclitaxel in the core fluid solution is about 20 μg/ml.
The shell fluid solution is prepared by mixing two functionalized biopolymers in chloroform: (a) Folate-functionalized polyethylene glycol), or Folate-PEG, with a molar weight of 5,000 Da; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5000 Da and a Mw/Mn=1.5. The weight percent contents of Folate-PEG and PCSH are about 0.30 wt % each.
Capsules with an average diamter in the range of about 0.250 μm to about 1 μm are generated, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates are about 0.050 and about 0.300 ml/h, respectively, and the external voltage is about 7.5 kV.
Capsules with a paclitaxel loading in the range of about 0.01 to about 0.2% by weight of the added polymer are made by adjusting the concentration of the core fluid solution.
Specific Example 22 Particles with PaclitaxelThe therapeutic solution is prepared by mixing Paclitaxel in dichloromethane. The concentration of Paclitaxel in the therapeutic solution is about 1,000 μg/mL.
A biopolymer solution is prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2,000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH are about 0.3 wt % and about 0.3 wt %, respectively.
The therapeutic and biopolymer solutions are mixed to form a homogenous solution. Particles with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flowrates is in the range of about 0.050 to about 0.300 ml/h, and the external voltage is about 7.5 kV.
Particles with a Paclitaxel loading in the range of about 0.01 and 25% by weight of the added polymer may by adjusting the concentration of the core fluid solution.
Specific Example 23 Encapsulation of PaclitaxelThe core fluid solution is prepared by mixing Paclitaxel in DMSO and about 10 wt % of γ-cyclodextrine in about 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of Paclitaxel in the core fluid solution is about 20 μg/mL.
The shell fluid solution is prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively; and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH are about 0.3 wt % each.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates are about 0.050 and about 0.300 ml/h, respectively, and the external voltage is about 7 kV.
Capsules with a Paclitaxel loading in the range of about 0.01 to about 0.2% by weight of the added polymer may be made y adjusting the concentration of the core fluid solution.
Specific Example 24 Encapsulation of Gold NanoparticlesThe core fluid solution is prepared by mixing an aqueous solution of gold nanoparticles, or NP-Au and a 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of NP-Au in the core fluid solution is in the range of about 1,000 to about 500 μg/mL.
The shell fluid solution is prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH are about 0.30 wt % and about 0.30 wt %, respectively.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates are about 0.050 and about 0.150 ml/h, respectively, and the external voltage is about 8 kV.
Capsules with a gold nanoparticle loading in the range of about 0.01 to about 11% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 25 Encapsulation of Silver NanoparticlesThe core fluid solution was prepared by mixing an aqueous solution of silver nanoparticles, or NP-Ag and a 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of NP-Ag in the core fluid solution was in the range of about 1,000 to about 500 μg/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, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH were about 0.30 wt % and about 0.30 wt %, respectively.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were in the range of about 0.050 to about 0.150 ml/h, respectively, and the external voltage was about 8 kV.
Capsules with a silver nanoparticle loading in the range of about 0.01 to about 11% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 26 Encapsulation of Palladium NanoparticlesThe core fluid solution was prepared by mixing an aqueous solution of palladium nanoparticles, or NP-Pd and a 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of NP-Pd in the core fluid solution was in the range of about 1,000 to about 500 μg/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, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; and (b) poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA and PCSH were about 0.30 wt % and about 0.30 wt %, respectively.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm were produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were in the range of about 0.050 to about 0.150 ml/h, respectively, and the external voltage was about 8 kV.
Capsules with a palladium nanoparticle loading in the range of about 0.01 to about 11% by weight of the added polymer may be made by this method by adjusting the concentration of the core fluid solution.
Specific Example 27 Particles with Paclitaxel and Estradiol Functional GroupsThe therapeutic solution is prepared by mixing Paclitaxel in dichloromethane. The concentration of Paclitaxel in the therapeutic solution is about 1,000 μg/mL.
A biopolymer solution is prepared by mixing two functionalized biopolymers in 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, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of EST-PEG and PCSH are about 0.30 wt % each.
The therapeutic and biopolymer solutions are mixed to form a homogenous solution. Particles with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flowrate is in the range of about 0.050 to about 0.300 ml/h range, and the external voltage is about 7.5 kV.
Particles with a Paclitaxel loading in the range of about 0.01 to about 25% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 28 Capsules with Paclitaxel and Estradiol Functional GroupsThe core fluid solution is prepared by mixing Paclitaxel in DMSO and about 10 wt % of γ-cyclodextrine in about 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and 2 mM of CaCl2. The final concentration of Paclitaxel in the core fluid solution is about 20 μg/mL.
The shell fluid solution is 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, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of EST-PEG and PCSH are about 0.30 wt % each.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates are about 0.050 and about 0.150 ml/h, respectively, and the external voltage is about 8 kV.
Specific Example 29 Particles with Paclitaxel and Epidermal Growth Factor GroupsThe therapeutic solution is prepared by dissolving Paclitaxel in dichloromethane. The concentration of Paclitaxel in the therapeutic solution is about 1,000 μg/mL.
A biopolymer solution is prepared by mixing three functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5; and (c) Epidermal growth factor-functionalized poly(ethylene glycol), or EGF-PEG, with a molecular weight of 5,000 Da. The weight percent contents of PEG-b-PLA, PCSH and EGF-PEG are about 0.30 wt %, about 0.30 wt % and about 0.10 wt %, respectively.
The therapeutic and biopolymer solutions are mixed to form a homogenous solution. Particles with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the flowrates are in the range of about 0.050 to about 0.300 μml/h. The external voltage is about 7.5 kV.
Particles with a Paclitaxel loading in the range of about 0.01 to about 25% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 30 Capsules with Paclitaxel and Epidermal Growth Factor GroupsThe core fluid solution is prepared by mixing Paclitaxel in DMSO and about 10 wt % of γ-cyclodextrine in a about 10 mM Bis-Tris propane aqueous solution containing about 1 wt %of isopropanol and 2 mM of CaCl2. The final concentration of Paclitaxel in the core fluid solution is about 20 μg/mL.
The shell fluid solution is prepared by mixing three functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5; and (c) Epidermal growth factor-functionalized poly(ethylene glycol), or EGF-PEG, with a molecular weight of 5,000 Da. The weight percent contents of PEG-b-PLA, PCSH and EGF-PEG are about 0.30 wt % and about 0.10 wt %, respectively.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates are about 0.050 and about 0.300 ml/h, respectively, and the external voltage is about 7.5 kV.
Capsules with a paclitaxel loading in the range of about 0.01 to about 0.2% by weight of the added polymer are made by adjusting the concentration of the core fluid solution.
Specific Example 31 Encapsulation of pCMV-Luc PlasmidThe core fluid solution is prepared by mixing a buffer solution containing pCMV-Luc plasmid containing the cytomegalovirus (CMV) promoter of pcDNA3 inserted upstream to the firefly luciferase of the pGL2-basic vector plasmid with a 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentration of pCMV-Luc in the core fluid solution is in the range of about 1 to about 1000 μg/mL.
The shell fluid solution is prepared by mixing three functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The weight percent contents of PEG-b-PLA, PCSH are about 0.30 wt % and about 0.30 wt %, respectively.
Capsules with an average diameter in the range of about 0.250 μm to about 1 μm are produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates are about 0.050 and about 0.150 ml/h, respectively, and the external voltage is about 8 kV.
Capsules with a pCMV-Luc loading is in the range of about 0.01 to about 11% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 32 Encapsulation of 425 TNF-α DNA and Bovine Serum Albumin Rhodamine ConjugateThe core fluid solution was prepared by mixing 425 TNF-α DNA, and bovine serum albumin rhodamine conjugate, or BSA-rhodamine, in about 10 mM Bis-Tris propane aqueous solution containing about 1 wt% of isopropanol and about 2 mM of CaCl2. The final concentration of 425 TNF-α DNA and BSA-rhodamine in the core fluid solution were about 12.2 μg/mL, and about 200 μg/mL, respectively.
The shell fluid solution was prepared by mixing of EGF grafted to a fluorescein-labeled poly(ethylene glycol)-NHS biopolymer, or F-PEG-EGF, 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 produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates used were about 05 and about 0.30 ml/h, respectively, and the applied voltage was about 6,5 kV.
Capsules with a 425 TNF-α DNA loading in the range of about 0.001 to about 30% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 33 Encapsulation of TemozolomideThe core fluid solution was prepared by mixing temozolomide or TMZ in about 0.1 M acetate buffer solution. The final concentration of TMZ in the core fluid solution is about 10 μM.
The shell fluid solution is prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The contents of PEG-b-PLA, PCSH in the shell fluid solution were about 0.080 wt % and about 0.086 wt %, respectively.
Capsules with an average diameter of about 1.3 μm were produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates were about 0.05 and about 0.30 ml/h, respectively, and the applied voltage was about 8.0 kV.
Capsules with a temozolomide loading in the range of about 0.01 to about 2.4% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
Specific Example 34 Encapsulation of TNF-α Protein and Bovine Serum Albumin Fluorescein ConjugateThe core fluid solution was prepared by mixing TNF-α protein, and bovine serum albumin fluorescein conjugate, or BSA-fluor 594, in about 10 mM Bis-Tris propane aqueous solution containing about 1 wt % of isopropanol and about 2 mM of CaCl2. The final concentrations of TNF-α protein and BSA-fluor in the core fluid solution were about 345 μg/ml, and about 200 μg/mL, respectively.
The shell fluid solution is prepared by mixing two functionalized biopolymers in chloroform: (a) COOH-poly(ethylene glycol)-b-polylactide, or PEG-b-PLA, with a molecular weight of 2000-b-1940 Da using same nomenclature, respectively, and a Mw/Mn=1.2; (b) Poly(caprolactone)-SH, or PCSH, with a molecular weight of 5,000 Da and a Mw/Mn=1.5. The shell fluid solution is doped with a solution of magnetite particles with an average diameter of about 1.5 nm. The contents of PEG-b-PLA, PCSH and magnetite particles in the shell fluid solution were about 0.080 wt % and about 0.086 wt %, respectively.
Capsules with an average diameter of about 0.225 μm and about 0.550 μm were produced, but smaller capsules may be made by adjusting the process variables. Specifically, the core and shell fluid flow rates were about 0.050 and about 0.300 ml/h, respectively, and the applied voltage was about 7.0 kV.
Capsules with a TNF-α protein loading in the range of about 0.01 to about 23% by weight of the added polymer may be made by adjusting the concentration of the core fluid solution.
The examples given above are merely illustrative and are not meant to be 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 invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection 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 polymer sciences, molecular biology or related fields are intended to be within the scope of the appended claims.
The disclosures of all references and publications cited above are expressly incorporated by reference in their entireties to the same extent as if each were incorporated by reference individually.
Claims
1. A method of producing a capsule having at least one encapsulated agent, said method comprising:
- supplying a core fluid to an interior of a hollow tube;
- supplying a shell fluid flow;
- generating a shell fluid flow to an external wall of the hollow tube;
- subjecting the core fluid and the shell fluid to an electric potential so as to form a two-fluid electrified jet at an apex of the hollow tube;
- forming a capsule having a core region and a shell region at the apex of the hollow tube.
2. The method of claim 1, wherein the core fluid is supplied at a flow rate in the range of about 0.005 mL/hour to about 5 mL/hour.
3. The method of claim 2, wherein the core fluid flow rate is about 0.025 mL/h.
4. The method of claim 1, wherein the shell fluid is supplied at a flow rate in the range of about 0.05 mL/hour to about 5 mL/hour.
5. The method of claim 4, wherein the core fluid flow rate is about 0.075 mL/h.
6. The method of claim 1, further comprising the steps of:
- supplying a collector electrode; and
- depositing the formed capsules onto a surface of the collector electrode.
7. The method of claim 1, wherein the electric potential is a positive electrical bias.
8. The method of claim 7, wherein the positive electrical bias has a voltage in the range of about 5 kV to about 18 kV.
9. The method of claim 6, wherein the collector electrode is at ground electric potential.
10. The method of claim 6, further comprising the step of:
- supplying an extractor body such that the electric potential is biased at an electrical potential between the two-fluid electrified jet and the collector electrode.
11. The method of claim 1, further comprising the step of dispersing at least one species of a magnetite particle within the shell region of the capsule.
12. The method of claim 11, wherein the magnetite particle species if Fe3O4.
13. The method of claim 11, wherein the at least one species of magnetite particle has a size in the range of about 1 nm to about 300 nm.
14. The method of claim 1, further comprising the step of dispersing at least one species of a photonic sensitive nanoparticle within the shell region of the capsule.
15. The method of claim 14, wherein the species of photonic sensitive nanoparticle is a compound selected from the group consisting of silver, gold, palladium, and any combination thereof.
16. The method of claim 14, wherein the photosensitive nanoparticle species has a size i the range of about 1 nm to about 50 nm.
17. The method of claim 1, further comprising the step of dispersing a functional group on the outer perimeter of the shell of the capsule.
18. The method of claim 17, wherein the function group is an entity selected from the group consisting of hydroxyl groups, amino groups, carboxyl groups, carboxylic acid anhydride groups, mercapto groups, hydrosilicon groups, thio groups, carboxylic groups, amine groups, and any combination thereof.
19. The method of claim 17, wherein the functional group is capable of chemically or physically attaching to pancreatic cancer cells.
20. The method of claim 19, wherein the functional group is one or more compounds selected from the group consisting of kinase receptors, fibroblast growth factor receptor, EGF, TGF, VEGF-A, urokinase receptor, interleukin-4 receptor, retinoic acid receptor, heparin-binding EGF-like growth factor, HB-EGF, amphiregulin, epireguin, neuregulins, and functional equivalents thereof.
21. The method of claim 20, wherein the functional group has a content in the range of about 0 wt % to about 1 wt %.
22. The method of claim 17, wherein the functional group is capable of chemically or physically attaching to glioma cells.
23. The method of claim 21, wherein the functional group is epidermal growth factor and a functional equivalent thereof.
24. The method of claim 23, wherein the functional group has a content on the capsule in the range of about 0 wt % to about 0.02 wt %.
25. The method of claim 17, wherein the function group is capable of chemically or physically attaching to breast cancer cells.
26. The method of claim 25, wherein the functional group is estrogen and functional equivalents thereof.
27. The method of claim 26, wherein the functional group has a content on the capsule in the range of about 0.02 wt % to about 0.4 25 %.
28. The method of claim 17, wherein the functional group is capable of chemically or physically attaching to at least one of lymphoma, myeloma and leukemia cancer cells.
29. The method of claim 28, wherein the functional group is at least one compound selected from the group consisting of VEGR-2, tostumomab, antibodies capable of binding to a CD20 receptor, CD5, CD7, CD13, CD19, CD22, CD33, CD52, CD61, antimyeloperoxidase, and functional equivalents thereof.
30. The method of claim 29, wherein the function group has a content on the shell of the capsule in the range of about 0.0 wt % to about 1.0 wt %.
31. The method of claim 1, wherein the shell fluid is at least one biocompatible polymer.
32. The method of claim 31, wherein the at least biocompatible polymer is a polymer selected from the group consisting of poly(lactic acid), poly(lactic-co-glycolic acid), citosan, alkyl metacrylates, polycaprolactone, starch, polyethylene glycol, and copolymers resulting from combination thereof.
33. The method of claim 1, wherein the core fluid is at least one of a therapeutic agent and an imaging agent.
34. The method of claim 33, wherein the therapeutic agent is one or more biological materials selected from the group consisting of nucleic acid, double stranded DNA, single stranded DNA, double stranded RNA, single stranded RNA, peptide nucleic acid(PNA), antisense DNA, antisense RNA, small interfering RNA, proteins, lipids, carbohydrates, and combination thereof.
35. The method of claim 33, wherein the therapeutic agent is at least one abiological material.
36. The method of claim 33, wherein the therapeutic agent is suitable for a treatment a patient afflicted with a condition.
37. The method of claim 36, wherein the patient is afflicted with at least one condition selected from the group consisting of breast cancer, glioma, lymphoma, leukemia, prostate cancer, pancreatic cancer, carcinoma, sarcoma, mesothelioma, glioma, germinoma, and choriocarcinoma.
38. The method of claim 33, wherein the imaging agent is suitable for detecting a malignant cancer in a patient.
39. A capsule including at least one encapsulated agent produced by the method of claim 1.
40. The capsule of claim 39, wherein the encapsulate agent is at least one of a therapeutic agent and an imaging agent.
41. The capsule of claim 39, wherein the capsule comprises a shell region and a core region.
42. The capsule of claim 41, wherein magnetic nanoparticles are dispersed in the shell region.
43. The capsule of claim 41, wherein functional groups are dispersed on an outer surface of the shell region.
44. The capsule of claim 43, wherein the functional group have an affinity to attach chemically or physically to a target cell.
45. The capsule of claim 44, wherein the target cell is a malignant cancer cell selected from the group consisting of breast cancer, glioma, lymphoma, leukemia, prostate cancer, pancreatic cancer, carcinoma, sarcoma, mesothelioma, glioma, germinoma, and choriocarcinoma.
46. The capsule of claim 41, wherein the shell region includes pores.
47. The capsule of claim 46, wherein the pores have a size in the range of about 0.5 nm to about 2.0 μm.
48. The capsule of claim 39, wherein the therapeutic agent is at least one of a chemotherapeutic agent, radioactive isotope, and a chemotherapeutic enhancing drug.
49. The capsule of claim 39, wherein the therapeutic agent is a compound selected from the group consisting of anti-inflammatory compounds, anti-allergics, glucocorticoids, anti-infective agents, antibiotics, antifungals, antivirals, mucolytics, antiseptics, vasoconstrictors, wound healing agents, local anaesthetics, peptides, and proteins.
50. The capsule of claim 17, wherein the functional group is capable of targeting a subcellular target.
51. The capsule of claim 50, wherein the subcellular target is one or more components selected from the group consisting of endoplasmic reticulum, mitochondria, golgi apparatus, vacuole, nucleus, aerosome, centriole, cilium, glyoxysome, lysosome, melanosome, myofibril, nucleolus, peroxisome, actin, tubulin, plasma membrane, and ribosome, vesicles.
52. A capsule, comprising:
- a core region;
- a shell region such that the shell region includes pores having a size in the range of about 0.5 nm to about 2.0 nm;
- at least one entrapped an/or encapsulated agent; and
- a functional group dispersed on a surface of the shell region.
53. The capsule of claim 52, wherein the encapsulated and/or entrapped agent is at least one of a therapeutic agent and an imaging agent.
54. The capsule of claim 52, wherein magnetic nanoparticles are dispersed in the shell region.
55. The capsule of claim 52, wherein the functional group has an affinity to attach chemically or physically to a target cell.
56. The capsule of claim 55, wherein the target cell is a malignant cancer cell selected from the group consisting of breast cancer, glioma, lymphoma, leukemia, prostate cancer, pancreatic cancer, carcinoma, sarcoma, mesothelioma, glioma, germinoma, and choriocarcinoma.
57. The capsule of claim 53, wherein the therapeutic agent is at least one of a chemotherapeutic agent, radioactive isotope, and a chemotherapeutic enhancing drug.
58. The capsule of claim 57, wherein the therapeutic agent is a compound selected from the group consisting of anti-inflammatory compounds, anti-allergics, glucocorticoids, and anti-infective agent, antibiotics, antifungals, antivirals, mucolytics, antiseptics, vasoconstrictors, wound healing agents, local anaethetics, peptides, and proteins.
59. The capsule of claim 52, wherein the functional group is capable of targeting a subcellular target.
60. The capsule of claim 52, wherein the subcellular target is one or more components selected from the group consisting of endoplasmic reticulum, mitochondria, golgi apparatus, vacuole, nucleus, aerosome, centriole, cilium, glyoxysome, lysosome, melanosome, myofibril, nucleolus, peroxisome, actin, tubulin, plasma membrane, and ribosome, vesicles.
Type: Application
Filed: May 2, 2007
Publication Date: Aug 7, 2008
Inventors: Gustavo LARSEN (Denton, NE), Ruben SPRETZ (Lincoln, NE), Raffet VELARDE-ORTIZ (Lincoln, NE), David VU (Lincoln, NE), Luis NUNEZ (Chicago, IL)
Application Number: 11/743,609
International Classification: A61K 51/12 (20060101); A61K 49/18 (20060101); A61K 9/127 (20060101); A61P 35/00 (20060101); B01J 13/02 (20060101);
