KR20110039466A - Particle compositions with pre-selected cell internalization mode - Google Patents

Abstract

The present invention relates to a method comprising the steps of selecting a cell having a surface receptor, i) surface components capable of binding to or binding to a receptor, and ii) surface distribution of the surface components and shapes thereof for the cells selected. A method of making a particle composition having a preselected cell internalization mode, comprising obtaining a particle having a preselected shape effective for a preselected cell internalization mode.

Description

Particle composition having a preselected cell internalization mode {PARTICLE COMPOSITIONS WITH PRE-SELECTED CELL INTERNALIZATION MODE}

Reference to federally funded research

Some studies underlying the present invention have received federal funding from DOD (subsidies W81XWH-04-2-0035) and NASA (subsidies SA23-06-017). The United States government may have certain rights in this invention.

Technical field

The present invention generally relates to micro and nano size particle compositions and uses thereof, and more particularly to micro and nano size particle compositions having certain preselected cellular internalization modes.

Inclusion is a general term that defines the processes by which cells can absorb and / or release selected extracellular species such as molecules, viruses, particles, and microorganisms and direct them to specific organs in the cytoplasm. Inclusion can occur through a variety of pathways, including clathrin-mediated and cabeore-mediated inclusion, phagocytosis, clathrin-mediated and caveole-independent endocytosis. Specific endogenous pathways may vary depending on the size and nature of the extracellular material, see, eg, Conner S.D. and S.L. Schmid. 2003. Nature 422: 37-44. For example, caboleole-mediated endocytosis is facilitated by caboleole activation, plasma membrane incorporation of characteristic sizes 50-60 nm, and clathrin-mediated endothelial activity has a characteristic size of several hundred microns (100-500 nm). Induction of the formation of vesicles requires concentration of transmembrane receptors and their bound ligands on the plasma membrane, and phagocytosis of specific cell-surface receptors and signaling cascades and micrometer size (> 1 μm) It involves the formation of cell membrane protrusions that substantially surround the foreign material. Clathrin- and Cabeoole independent endocytosis may be associated with the formation of embedded vesicles of less than 100 nm.

Particle inclusion can be very important in many fields such as virology, drug and gene delivery and in nanotoxicology. See, eg, Marsh M. and A. Helenius. 2006. Cell 124: 729-40; Vasir JK, and V. Labhasetwar. 2006. Expert Opin.Drug Deliv. 3: 325-344; Oberdorster G., E. Oberdorster, J. Oberdorster. 2005. 113: 823-39.

For nano-sized particles, the most effective internalization mechanism, whether natural as an enveloping virus or artificial as a biomimetic particle, binds the corresponding molecule (receptor) where molecules (ligands) distributed over the particle surface are expressed across the cell membrane. It can be a receptor-mediated endocytosis that can be substantially bent to incorporate foreign material. See, eg, Marsh M. and A. Helenius. 2006. Cell 124: 729-40; Smith A.E., A. Helenius. 2004. Science 304: 237-42. These receptors can be collected at the site of infiltration by surface diffusion, otherwise no endocytosis occurs or occurs over a much longer time.

summary

One embodiment includes a) selecting a cell having a surface receptor; And b) surface components capable of affinity or binding to the receptor, and ii) surface distribution and shape of the surface components having a shape effective for a preselected cell internalization scheme for the selected cells. It provides a method for producing a particle composition having a pre-selected cell internalization mode, comprising the step of obtaining the particles.

Another embodiment is a method of treating or monitoring a physiological condition comprising administering an effective amount of a particle composition having a preselected mode of cell internalization to a subject in need of treatment or monitoring of the physiological condition.

Another embodiment is a particle composition having a preselected mode of cellular internalization.

Brief description of the drawings

1 schematically illustrates particles having an elliptical cross section. The particles carry ligand molecules on their surface that can interact with receptor molecules on the cell membrane surface.

FIG. 2 shows the change of elliptic cylindrical particles representing a half-wrapping time 0.5τ _w and a wrapping length ratio x _max / R ₁ as a function of aspect ratio (Γ), with R ₂ fixed at 50 nm. In the range from 0.9 to 4.

3 is at least half a graph showing the value Γ Γ _min for a fixed R ₂ corresponding to the wrapping time, as a function of R _2. 3 is also a graph showing the half lapping time for Γ _min as a function of R ₂ .

FIG. 4 shows the change of elliptic cylindrical particles representing half lapping time 0.5τ _w and lapping length ratio x _max / R ₁ as a function of aspect ratio (Γ), from 0.9 to a fixed volume (R _s = 50 nm). 4 ranges.

details

Related literature

The following research papers and patent documents, the entire contents of which are incorporated herein by reference, may be useful in understanding the present invention:

1) PCT publication WO 2007/120248 published October 25, 2007;

2) PCT publication WO 2008/041970, published April 10, 2008;

3) PCT publication WO 2008/021908, published February 21, 2008;

4) US Patent Application Publication No. 2008/0102030, published May 1, 2008;

5) US Patent Application Publication 2003/0114366;

6) US Patent Application 12 / 034,259, filed February 20, 2008;

7) US patent application 12 / 110,515, filed April 28, 2008;

8) Tasciotti et al, Nature Nanotechnology, Vol. 3, 151-158, 2008;

9) Decuzzi and Ferrari, Biomaterials 28, 2007, 2915-2922;

10) Decuzzi and Ferrari, Biophysical Journal, 94, 2008, 3790-3797.

Justice

Unless otherwise stated, singular forms mean one or more.

Unless stated otherwise, the terms "inclusion" and "inclusion" mean "receptor mediated inclusion" and "receptor mediated inclusion", respectively.

"Microparticle" means a particle having a maximum maximum of 1 μm to 1000 μm or in some cases from 1 micron to 100 microns.

"Nanoparticle" means a particle having a maximum dimension of less than 1 micron.

By “phagocytosis” is meant the uptake of large (characteristic size greater than 2 microns) particles by specialized macrophages including macrophages, monocytes and neutrophils. Phagocytosis involves the formation of protrusions in the cell membrane, which substantially surround the outer particles.

"Receptor mediated endocytosis" or RME means internalization by cells of a particle in which a component such as a ligand capable of binding to a corresponding component (receptor) expressed over the cell membrane is distributed on its surface. RME involves bending of the cell membrane causing substantial internalization of the particles by the cells and complete lapping of the particles by the membrane. Particles internalized through RME may have a smaller characteristic size than particles undergoing internalization through phagocytosis. RME is not limited to phagocytes.

"Biodegradable" means a biocompatible polymer material or a material that can be dissolved or degraded in a physiological medium that can be degraded under physiological and / or chemical conditions by a physiological enzyme.

Contents

We have recognized particle shapes important for receptor mediated inclusion. Accordingly, the present invention includes the steps of selecting a target cell and obtaining a particle having a surface component having affinity for or binding to a surface receptor on the surface of the target cell on the surface thereof. Provided are methods for preparing a particle composition having a selected cell internalization mode. The surface distribution of the surface components on the particles and the shape of the particles are the distributions and shapes that make it effective for the preselected mode of cell internalization for the selected cells.

The preselected cell internalization mode may be selected from the “involvement” or non-incorporation mode. “Incorporation” is when the particle can be completely enclosed and substantially internalized by the cell membrane via receptor mediated inclusion. By way of the term “non-inclusion” is meant when the particle can only be partially wrapped by the cell membrane.

In some embodiments, the preselected mode of cell internalization may be incomplete or partial inclusion. "Frustrated endocytosis" refers to the manner in which the particles are only partially wrapped by the cell membrane and are not internalized by the cells.

Particles having a preselected mode of cellular internalization can be used for the treatment and / or monitoring of physiological conditions. In this case, in the body of a subject, such as a mammal, preferably a human, it selects a target site with cells having surface receptors on its surface and has a preselected internalization scheme for cells of this target portion. An effective amount of a composition comprising particles may be administered to the subject. The physiological condition may be a disease such as cancer or inflammation, for example.

In general, the selected cell can be any type of cell having a surface receptor on its surface. In many embodiments, the selected cell can be a mammalian cell, such as a human cell.

The internalization mode that is specifically chosen may depend on the intended use of the particles. For example, when the particles contain a substance such as a contrast agent or therapeutic agent that is desired to be delivered inside the cell, the inclusion method is preferred. On the other hand, if the particle contains a substance that is not intended for delivery into the cell, a non-inclusion or incomplete inclusion mode may be desirable. An example of such a situation may be the multistage delivery vehicle disclosed in PCT publication WO 2008/021908, wherein the first stage vehicle particles containing the second stage particles therein are not internalized by endothelial cells and are recognized at target sites in the endothelium. And are attached. After attachment, the first stage particles can release the second stage particles.

In many embodiments, the selected cell may be a non-phagocytic cell, i.e., a cell unable to perform phagocytosis. Examples of phagocytes, ie cells that perform phagocytosis, include neutrophils, monocytes and macrophages.

In some embodiments, the selected cell can be an endothelial cell, such as a vascular endothelial cell, and the target site can be a vascular structure site, such as a shared vascular structure, angiogenic vascular structure, or an reorganized vascular structure. In cells of shared vasculature, the surface receptor may be an angiopoietin 2 receptor; In angiogenic cells, the surface receptor may be an endothelial marker such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor or α _v β ₃ integrin; In reconstituted vasculature cells, the surface receptors are cancer-associated cell adhesion molecule 1 (CEACAM1), endothelin-B receptor (ET-B), vascular endothelial growth factor inhibitor Gravin / AKAP12, protein kinase A and protein kinase C Scaffold protein.

The surface components of the particle surface can be complementary to the receptors on the surface of the selected cell. Surface components can be, for example, antibodies, aptamers or ligands that have affinity for, or can bind to, receptors on selected cell membrane surfaces. In some embodiments, surface components can include components specific for a receptor on a selected cell membrane surface. In some embodiments, the surface components can include components that are not specific for the receptor on the selected cell membrane surface. In other embodiments, the surface components can include specific and nonspecific components.

Particles having a preselected internalization mode may be part of a composition that may also include particles that do not have a preselected internalization mode. Particles having a preselected internalization mode may account for at least 10% or at least 25% or at least 50% or at least 75% or at least 90% by number of the total number of particles in the composition.

In many embodiments, the particles can be non-spherical particles. In some embodiments, the particles can be particles having a circular cross section. In some embodiments, the particles can be elliptical particles. In another embodiment, the particles can be cylindrical particles having an elliptical cross section in a direction perpendicular to the axis of the cylinder.

In some embodiments, the maximum characteristic size of a particle, such as the long axis length of the particle having an elliptical cross section, is less than 2 microns or less than 1 micron or less than 800 nm or 5 nm to 500 nm or 5 nm to 800 nm, 5 nm to 1 Micron or 10 nm to 1 micron or 10 nm to 800 nm or 10 nm to 500 nm or 20 nm to 1 micron or 20 nm to 800 nm or 20 nm to 500 nm or 50 nm to 1 micron or 50 nm to 800 nm or 50 nm to 500 nm.

Preferably, the maximum characteristic size of the particles is substantially smaller than the characteristic size of the selected cell. The maximum characteristic size of the particles is at least 3 times or at least 5 times or at least 10 times or at least 20 times or at least 30 times or at least 50 times or at least 100 times or at least 200 times or at least 300 times the characteristic size of the selected cells or It may be at least 500 times or at least 100 times smaller. The characteristic size of the selected cells can vary from about 5 microns to about 40 microns or from about 10 microns to about 30 microns.

In some embodiments, the particles obtained may be particles having a convex surface that retains the local surface density and local curvature of the effective component (e.g., ligand) of the particle via receptor mediated inclusion, which is the local surface of the component. It means that the local curvature κ for density is less than the maximum inclusion curvature κ _max . The maximum endogenous curvature may vary depending on the binding energy between the components on the particle surface and the receptors on the surface of the selected cell membrane, the binding energy and particle components of the selected cell membrane and the surface density of the receptors of the selected cell membrane. The method of evaluating the maximum nesting curvature is described below.

In some embodiments, the local surface density of the component at the convex surface may be greater than the surface density of the component at other portions of the particle surface, so that the surface distribution of the surface component on the particle surface may be nonuniform.

In particles having an elliptic cross section, obtaining particles having a preselected mode of cell internalization may involve obtaining particles having an aspect ratio corresponding to the selected mode.

For example, FIG. 1 shows an elliptical cross section of elliptical cylindrical particles that interact with the membrane through specific ligand receptor binding. The surface density of the ligand on the particle surface is m _l, and the surface density of the receptor on the cell membrane surface is m _r .

The elliptical cross section may be characterized by R ₁ and R ₂ , which are half lengths of the elliptic axis. The aspect ratio of the ellipsoid can be defined as Γ = R ₁ / R ₂ . The cross-sectional area of the particles is A = πR ₁ R ₂ = πΓR ₂ ² = πR _s ² , where R _s is the radius of the particles with the same circular cross section as in the case of elliptic particles.

The geometry of the particles is R ₁ and R ₂ ; Γ and R ₁ ; Can be completely determined by the number of parameter pairs, such as Γ and R ₂ , Γ and R _s or Γ and V, where V is the volume of the particle.

In this case, when the geometry of the particle is determined by Γ and R ₂ , the shape parameter that determines the preselected internalization scheme may be the aspect ratio Γ for the particular R ₂ .

Particles with elliptical cross-sections are defined as Γ '= (R ₂ κ _max ) ^-1/2 , where κ _max is the maximum nesting, if the aspect ratio of the particle to the uniaxial half-length of the particle is less than the first threshold, Action curvature), it may be in a "non-inclusion" manner.

Particles may be in an incomplete nesting mode if the aspect ratio of the particles to a particular half length of the uniaxial is below the second threshold (Γ "= R ₂ κ _max ).

Particles may be nested when the aspect ratio of the particles to a particular half-length of the uniaxial is greater than the first threshold Γ 'and less than the second threshold Γ ".

The maximum containment curvature κ _max can be defined as the inverse of the calculated minimum containment radius for cells selected for spherical particles or particles having a circular cross section and the same ligand as the non-spherical particles.

The minimum nesting radius, and therefore the maximum nesting curvature, can be evaluated using the following equation (1):

(여기서, K_d ^2D는 세포/입자 계면에서 상호작용하는 리간드-수용체의 평형 해리 상수)이다. K_d ^2D는 관계식 K_d ^2D = K_d/h(여기서, K_d는 예컨대 실험적으로 용액에서 측정한 동일한 리간드-수용체 쌍의 평형 해리 상수이고, h는 리간드-수용체 부위가 제한되는 제한 영역의 두께임)으로부터 추산될 수 있다. 많은 경우, h는 대략 10 nm일 수 있다.In this equation, C is the ligand-receptor binding energy for κ _B T, where κ _B is the Boltzmann constant and T is the temperature (absolute temperature) of the cell or target site, expressed in Kelvin. C depends on the specific ligand-receptor pair. Specifically, C =

Where K _d ^2D is the equilibrium dissociation constant of the ligand-receptor interacting at the cell / particle interface. K _d ^2D is the relationship K _d ^2D = K _d / h, where K _d is the equilibrium dissociation constant of the same ligand-receptor pair measured in solution, for example, and h is the thickness of the restriction region at which the ligand-receptor site is restricted. Can be estimated from In many cases h can be approximately 10 nm.

B is the binding energy factor of the cell membrane, which can be determined as detailed in Hochmuth, R.M., J. Biomech., 33: 15-22, 2000.

If the particles do not interact with the cells, the average surface density m _r on the receptor can be determined using methods known to those skilled in the art. For example, radiolabeled monoclonal antibodies complementary to the receptor as described above for intercellular adhesion molecule 1 receptor in Pannes J., et al. Am. J. Physiol. 1995; 269 (6Pt2): H1955-64. M _r can be determined in vivo. Alternatively, m _r can be determined using fluorescently labeled monoclonal antibodies complementary to the receptor. Such fluorescently labeled monoclonal antibodies can be antibodies labeled with phycoerythrin disclosed in US Pat. No. 4,520,110.

m _l is a local surface density of the ligand, m _l, it can be changed by changing the size of the control by and / or ligand molecules for surface functionalization conditions for the particles. The actual surface density of the ligand on the particles can be demonstrated experimentally using radiolabeled corresponding molecule or flow cell assays in radioactivity assays.

For particles with a uniform surface distribution of ligands, the local and average surface densities can be the same.

In certain embodiments, the minimum nesting radius and thus the maximum nesting curvature are evaluated as disclosed in US Patent Application No. 12 / 034,259, "Endocytotic particles", filed February 20, 2008, the entire contents of which are incorporated herein by reference. Can be.

FIG. 2 shows the variation of elliptic cylindrical particles exhibiting a half lapping time 0.5τ _w and a lapping length ratio x _max / R ₁ as a function of aspect ratio (Γ), the entire contents of which are incorporated herein by reference. In the range from 0.9 to 4 for R _s = 50 nm evaluated using the theoretical model disclosed in Decuzzi and Ferrari, Biophysical Journal, 94, 2008, 3790-3797. χ _max is the projection of the particle wrapping length across the x-axis of FIG. 1. At χ _max / R ₁ = 1, complete lapping occurs at τ _w time. In FIG. 2, in the case of Γ <Γ ' _cr (about 0.9 in this case), lapping is not possible even by inducing an infinitely large τ _w and zero ratio χ _max / R ₁ . At the median Γ of Γ ' _cr <Γ <Γ " _cr , the χ _max / R ₁ ratio is always a unit that means complete internalization of the particles and 0.5 τ _w increases almost linearly with Γ. In Γ " _cr , χ _max / R ₁ decreases, reaching a minimum and then steadily increasing as Γ increases.

3 is at least half a graph showing the value Γ Γ _min for a fixed R ₂ corresponding to the wrapping time, as a function of R _2. 3 is also a graph showing the anti-lapping time for Γ _min as a function of R ₂ . The same data is shown in Table 1 together with Γ ' _cr and Γ " _cr .

R ₂ , nm Γ _min (0.5 τ _w ) _min , sec Γ ' _cr Γ " _cr 38.35 (= R _min )) One ∞ One One 50 0.96 41.60 0.87 1.30 75 0.80 78.33 0.71 1.95 100 0.68 123.4 0.62 2.61 150 0.56 243.7 0.50 3.91 300 0.40 823.6 0.36 7.82 500 0.32 2105.2 0.28 13.0

As particle size R ₂ increases, aspect ratio Γ _min with minimum internalization time decreases from 1 (R ₂ = R _min ) to 0.3 (R ₂ = 500 nm).

In some cases, two parameters representing the geometry of the elliptic cylindrical particles can be Γ and R _s .

In this case, the shape parameter that determines the preselected cell internalization scheme may be an aspect ratio Γ for a specific value R _s .

Particles with elliptical cross-sections have a Γ ₁ '= (R ₂ κ _max ) ^-2/3 , where κ _max is the maximum nesting curvature, provided that the aspect ratio of the particle to the uniaxial half-length of the particle is less than the first threshold. Can be in a "non-inclusion" mode.

If the aspect ratio of the particles to a particular half-length of the uniaxial is below the second threshold (Γ ₁ "= R ₂ κ _max ) ^2/3 particles may be incomplete nesting mode.

Particles may be nested when the aspect ratio of the particle with respect to a particular half length of the uniaxial is greater than the first threshold Γ ₁ ′ and less than the second threshold Γ ₁ ″.

4 shows the change of elliptic cylindrical particles exhibiting a half lapping time 0.5τ _w and a lapping length ratio x _max / R ₁ as a function of aspect ratio (Γ), the entire contents of which are incorporated herein by reference ( Range from 0.9 to 4 for fixed volumes (R _s = 50 nm) evaluated using the theoretical models disclosed in Decuzzi and Ferrari, Biophysical Journal, 94, 2008, 3790-3797.

Types of particles

The type of particles having a preselected internalization mode is not particularly limited. For example, the particles can be liposomes, polymeric particles, silicon- and silica-based particles, quantum dots, gold nanoshells, dendrimers or viral particles.

In some embodiments, the particles can be prepared to have shapes that are effective for the preselected internalization scheme. In some embodiments, the particles having a shape effective for a preselected internalization scheme can be selected from particle pools that can have a wide range of shapes and / or size distributions. The selection from the particle pool can be made using a Zetasizer ™ Nano Series instrument from Malvern Instruments, Inc., Worcestershire, UK, which can measure the geometric dimensions of the particles.

Particles can be produced using a number of methods. In general, manufacturing methods that provide control over the size and shape of the particles may be desirable.

In some embodiments, the particles can be prepared by top-down microfabrication or nanofabrication methods such as photolithography, electron beam lithography, X-ray lithography, far ultraviolet lithography or nanoprint lithography. The advantage of using a top down manufacturing method is that in this way particles with uniform dimensions can be produced on a large scale.

The particles may carry a target component such as a ligand, aptamer or antibody on the surface. For example, the ligand can be chemically bound to an appropriate reactor on the particle surface. Protein ligands can be bound to amino reactive groups and thiol reactive groups under conditions effective to form thioether or amide bonds, respectively. Methods for attaching antibodies or other polymeric binders to inorganic or polymeric supports are described in detail, for example, in Taylor, R., Ed., Protein Immobilization Fundamentals and Applications, pages 109-110 (1991).

Non-uniform surface distribution of the surface components can be obtained when the particles are produced, for example, by top-down microfabrication or nanofabrication methods. For example, the substrate from which the particles are made can be patterned using a coating that resists ligand deposition such that the particles have two or more different surfaces, one that is resistant to ligand deposition and the other that is not. Subsequently exposing the substrate to a solution containing the ligand may result in particles with a heterogeneous distribution of ligand upon peeling from the substrate.

In some embodiments, the particles can have one or more channels connecting the surface and the reservoir. In some embodiments, the reservoir and channel can be voids on the particle body. In such cases, the particles may comprise porous or nanoporous materials. The pores of the porous or nanoporous material can be controlled to achieve a desired activator loading and / or a desired release rate. The nanoporous material having controllable pore size may be an oxide material such as SiO ₂ , Al ₂ O ₃ or TiO ₂ . The preparation of nanoporous oxide particles, also known as solgel particles, is described, for example, in Paik JA et. al. J. Mater. Res., Vol. 17, Aug 2002. The nanoporous material with controllable pore size may also be nanoporous silicon. For details on the preparation of nanoporous silicon particles, see, eg, Cohen MH et. Ah Biomedical Microdevices 5: 3, 253-259, 2003.

In some other embodiments, the particles may not contain any channels. Such particles may include, for example, biodegradable materials. For example, the particles may be formed of metals such as iron, titanium, gold, silver, platinum, copper and materials such as alloys and oxides thereof. Biodegradable materials may also be biodegradable polymers such as polyorthoesters, polyanhydrides, polyamides, polyalkylcyanoacrylates, polyphosphazenes and polyesters. Exemplary biodegradable polymers are disclosed, for example, in US Pat. Nos. 4,933,185, 4,888,176 and 5,010,167. Specific examples of such biodegradable polymeric materials include poly (lactic acid), polyglycolic acid, polycaprolactone, polyhydroxybutyrate, poly (N-palmitoyl-trans-4-hydroxy-L-proline ester) and poly (DTH Carbonates).

In certain embodiments, the particles can themselves be active agents.

Active agent

The active agent can be a therapeutic compound and / or contrast agent. The choice of specific active agent depends on the desired application.

The therapeutic agent may be a physiological or pharmacologically active substance capable of exerting some biological effect on the incomplete vasculature of a subject such as a mammal or a human. The therapeutic agent can be an inorganic or organic compound, including peptides, proteins, nucleic acids, and small molecules. Therapeutic agents include constant molecules, molecular complexes, pharmacologically acceptable salts such as hydrochloride, hydrobromide, sulfate, laurate, palmitate, phosphate, nitrite, nitrate, borate, acetate, maleate, tartarate, oleate It can be in various forms such as acid salts, salicylates, and the like. For acidic therapeutic agents, salts of metals, amines or organic cations such as quaternary ammonium can be used. Derivatives of drugs such as bases, esters and amides can also be used as therapeutic agents. The water-insoluble therapeutic agent can be used in the form of its water-soluble derivative or as its base derivative, which in any case or by its transfer is converted to an enzyme and hydrolyzed to its original therapeutically active form by the body's pH or other metabolic processes do.

Therapeutic agents may be chemotherapeutic agents, immunosuppressants, cytokines, cytotoxic agents, nucleolytic compounds, radioisotopes, receptors and prodrug activating enzymes that may be naturally occurring or prepared by synthetic or recombinant methods or combinations thereof. Can be.

Such as vinca alkaloids (eg vinblastine and vincristine), anthracyclines (eg doxorubicin and daunorubicin), RNA transcription inhibitors (eg actinomycin-D) and microtubule stabilizing drugs (eg paclitaxel) Drugs affected by conventional multiple drug resistance may be particularly useful as therapeutic agents.

Cancer chemotherapeutic agents may also be preferred therapeutic agents. Useful cancer chemotherapeutic drugs include nitrogen mustard, nitrosourea, ethyleneimine, alkane sulfonate, tetrazine, platinum compounds, pyrimidine analogs, purine analogs, metabolic compounds, folic acid analogs, anthracyclines, taxanes, vinca alkaloids, topo Isomerase inhibitors and hormones. Exemplary chemotherapeutic drugs include actinomycin-D, alkeran, Ara-C, anastrozole, asparaginase, BiCNU, bicalutamide, bleomycin, busulfan, capecitabine, carboplatin, carboplatinium , Carmustine, CCNU, chlorabusil, cisplatin, cladribine, CPT-11, cyclophosphamide, cytarabine, cytosine aramidoside, cytoxan, dacarbazine, dactinomycin, daunorubicin, dexra Foot acid, docetaxel, doxorubicin, DTIC, epirubicin, ethyleneimine, etoposide, phloxuridine, fludarabine, fluorouracil, flutamide, potemustine, gemcitabine, herceptin, hexamethylamine, hydride Loxyurea, Idarubicin, Iphosphamide, Irinotecan, Romustine, Mechloretamine, Melphalan, Mercaptopurine, Methotrexate, Mitomycin, Mitotan, Mitoxantrone, Oxaliplatin, Paclitaxel, Pamilonate, Pentostatin , Plicama Leucine, procarbazine, rituximab, steroid, streptozosin, STI-571, streptozosin, tamoxifen, temozolomide, teniposide, tetrazine, thioguanine, thiotepa, tomudex, topotecan, threosulfane, Trimetrexate, vinblastine, vincristine, vindesine, vinorelbine, VP-16 and xceloda.

Useful cancer chemotherapeutic drugs also include alkylating agents such as thiotepa and cyclophosphamide; Alkyl sulfonates such as busulfan, impprosulfan and pifosulfan; Aziridine, such as benzodopa, carbocon, meturedopa and uredopa; Ethyleneimine and methyllamelamine, such as altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolmelamine; Nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphamide, esturamustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nocheviehin, fensterrin, Prednismustine, trophosphamide, uracil mustard; Nitroreas such as cannustin, chlorozotocin, potemustine, lomustine, nimustine and rannimustine; Antibiotics such as arachinomycin, actinomycin, otramycin, azaserine, bleomycin, cocktinomycin, calicheamicin, carabicin, carminomycin, calginophylline, chrominomycin, dactinomycin, daunorucin Bisine, detorrubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, edambicin, marcelomycin, mitomycin, mycophenolic acid, nogalamycin, olibomycin, Peplomycin, fort pyromycin, puromycin, kelamycin, rhorubicin, streptonigrin, streptozosin, tubercidine, ubenymex, ginostatin, and zorubicin; Metabolic agents such as methotrexate and 5-fluorouracil (5-FU); Folic acid analogs such as denophtherine, methotrexate, putrophtherin and trimetrexate; Purine analogs such as fludarabine, 6-mercaptopurine, thiamipurine and thioguanine; Pyrimidine analogs such as ancitabine, azacytidine, 6-azauridine, carmopur, cytarabine, dideoxyuridine, doxyfluridine, enositabine, phloxuridine and 5-FU; Androgens, such as calusosterone, drostatanolone propionate, epithiostanol, renpythiostane and testosterone; Anti-adrenal such as aminoglutetidemide, mitotan and trilostane; Folic acid supplements such as proline acid; Aceglaton; Aldophosphamide glycosides; Aminoebulinic acid; Amsacrine; Best La Vucil; Bisantrene; Edatraxate; Depopamine; Demecolsin; Diaziquone; Elponnitine; Elliptinium acetate; Etogluside; Gallium nitrate; Hydroxyurea; Lentinane; Rodidamine; Mitoguazone; Mitoxantrone; Fur mall; Nitracrine; Pentostatin; Penammet; Pyrarubicin; Grape filinic acid; 2-ethylhydrazide; Procarbazine; PSK®; Lakamic acid; Sizofran; Spirogermanium; Tenuazone acid; Triazicones; 2,2 ', 2 "-trichlorotriethylamine; urethane; bindesin; dacarbazine; mannosemusin; mitobronitol; mitolactol; fifobroman; spinytocin; arabinoxide (" Ara-C "); Cyclophosphamide; thiotEPa; taxoids such as paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, NJ) and docetaxel (TAXOTERE®, Rhone-Poulenc Rorer, Antony, France); Ambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); phosphamide; mitomycin C; Mitoxantrone; vincristine; vinorelbine; nabelbin; novantron; teniposide; daunomycin; aminopterin; xceloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoro Methylornithine (DMFO); Litinoic acid; Esperamicin; Capecitabine; and Pharmaceutically acceptable salts, acids or derivatives of any of the groups, such as tamoxifen, raloxifene, aromatase inhibitory 4 (5) -imidazole, 4-hydroxytamoxifen, trioxyphene, keoxyphene, Antiestrogens, including onnapristone and toremifen; and androgens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts of any of the above Or anti-hormonal agents that act to modulate or inhibit hormonal action on tumors, such as derivatives.

Cytokines can also be used as therapeutic agents. Examples of such cytokines are lymphokine, monocaine and conventional polypeptide hormones. Cytokines include growth hormones such as human growth hormone, N-methionyl human growth hormone and bovine growth hormone; Parathyroid hormone; Thyroxine; insulin; Proinsulin; Relaxine; Prorelaxin; Glycoprotein hormones such as follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH) and luteinizing hormone (LH); Liver growth factor; Fibroblast growth factor; Prolactin; Placental lactogen; Tumor necrosis factor-α and -β; Mullerian inhibitors; Mouse gonadotropin associated peptide; Inhivin; Actibin; Vascular endothelial growth factor; Integrin; Thrombopoietin (TPO); Nerve growth factors such as NGF-β; Platelet growth factor; Converting growth factors (TGF) such as TGF-α and TGF-β; Insulin-like growth factor-I and -II; Erythropoietin (EPO); Osteoinduction factors; Interferons such as interferon-α, -β and -γ; Colony stimulating factors (CSFs) such as macrophage-CSF (M-CSF); Granulocyte-macrophage-CSF (GM-CSF); And granulocyte-CSF (GCSF); IL-I, IL-Ia, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12, IL- Interleukin (IL) such as 15; Tumor necrosis factor such as TNF-α or TNF-β; And other polypeptide factors including LIF and kit ligand (KL). As used herein, the term cytokine includes proteins derived from natural sources or from biologically active equivalents of recombinant cell culture and native array cytokines.

The contrast agent may be a substance that can provide contrast information about the targeted site in the body of an animal, such as a mammal or a human. The contrast agent may comprise a magnetic material such as iron oxide for magnetic resonance imaging. In optical contrast, the active agent can be, for example, semiconductor nanocrystals or quantum dots. In optical coherence tomography, the contrast agent may be a metal, such as gold or silver, nanocage particles. The contrast agent may also be an ultrasonic contrast agent such as micro or nanobubbles or iron oxide micro or nanoparticles.

Composition

Particles having a preselected internalizing cellular mode for a particular cell type may be part of a composition, such as a pharmaceutical composition. Such compositions may be suspensions comprising the particles disclosed above for use in administering a therapeutic or contrast agent. To form a suspension, the particles can be suspended in an aqueous medium at a selected concentration. The optimal concentration may vary depending on the nature of the particles (eg, available properties), the type of therapeutic product and the mode of administration. For example, compositions for oral administration may be relatively viscous and may contain high concentrations of particles (eg,> 50%). The solution for bolus injection preferably contains a relatively concentrated particle suspension (eg 10-50%) but is not concentrated to have a viscosity significantly higher than that of saline (relieving the need for needles with large holes). To minimize). Solutions used for continuous intravenous infusion may generally contain relatively low concentrations of particles (eg, 2-10% suspension) because relatively large volumes of fluid are administered.

The particles can be suspended in a number of suitable aqueous carrier vehicles. Suitable pharmaceutical carriers may be carriers which are nontoxic to the subject to be administered at the dosages and concentrations employed and are compatible with the other ingredients in the formulation. Examples of suitable carrier vehicles include, but are not limited to, water, saline, Ringer's solution, dextrose solution and 5% human serum albumin. Suspensions for use in injectable preparations are preferably isotonic with the blood of the subject. Generally, carriers include substances that enhance isotonicity and chemical stability, such as buffers and preservatives, and low molecular weight (less than about 10 residues) polypeptides, proteins, amino acids, carbohydrates including glucose or dextrins, chelating agents such as EDTA or other excipients. It may contain small amounts of additives such as

Prior to administration to the subject, the particle suspension can be sterilized by any suitable sterilization method. Particles made from thermostable materials can be heat sterilized, for example using an autoclave. Particles made from materials that are not thermally stable can be sterilized by passing commercially available sterilization filters. Of course, filtration can only be used if the particles are smaller than the pores of the sterile filter.

The particles can be administered to a subject in need of treatment through a number of suitable methods of administration. The specific method used for the specific application can be determined by the attending physician. For example, the particles can be administered by one of the following routes: topical, parenteral, aspiration, oral, vaginal and anal. Intravascular administration, including intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) infusions, may be particularly preferred.

Intravascular administration can be topical or systemic. Local vascular delivery can be used to bring the particles near the body part with known tumor or inflammation by the use of a guide catheter system such as a CAT-scan guide catheter. Common injections, such as bolus intravenous injection or continuous / trickle-feeding intravenous injection, are generally systemic injections.

The particles can be injected into the blood stream, circulated and localized at the target site. Preferably, the particles are injected into vascular tissue at the target site.

references

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While certain preferred embodiments have been mentioned so far, it will be understood that the invention is not so limited. Those skilled in the art will contemplate various changes to the disclosed embodiments and such changes fall within the scope of the present invention.

All publications, patent applications, and patents cited herein are hereby incorporated by reference in their entirety.

Claims (22)

a) selecting a cell having a surface receptor; And
b) particles having i) affinity for the receptor, or surface components capable of binding to the receptor, and ii) a surface distribution of the surface components and the shape thereof effective for the cell internalization scheme preselected for the selected cells. Obtaining Step
Method for producing a particle composition having a pre-selected cell internalization method, comprising. The method of claim 1, wherein the preselected internalization mode is selected from full nesting, frustrated nesting and non- nesting. The method of claim 1, wherein the cell is an endothelial cell. The method of claim 3, wherein the endothelial cells are vascular endothelial cells. The method of claim 4, wherein the receptor is an angiogenic vascular structure receptor, a coopted vascular structure receptor or a renormalized receptor. The method of claim 1, wherein the cells are non-phagocytic cells. The method of claim 1, wherein the surface component is a ligand capable of binding to a receptor. The method of claim 1, wherein said obtaining step comprises particle preparation. The method of claim 8, wherein the manufacture comprises preparation by a top-down process. The method of claim 8, wherein the preparation comprises disposing the components on the surface of the particle. The method of claim 1, wherein said obtaining step comprises selecting particles from the particle population. The method of claim 1, wherein the particles comprise an active agent. The method of claim 12, wherein the active agent comprises a contrast agent or a therapeutic agent. The method of claim 1, wherein the particles are nanoparticles. The method of claim 1, wherein the particles are nonspherical particles. The method of claim 1, wherein the particles are particles that do not have a circular cross section. The method of claim 1, wherein the particles are elliptical particles. 18. The method of claim 17, wherein the ellipsoidal particles have an aspect ratio effective for a preselected cell internalization mode for the selected cells. 19. The method of claim 18, further comprising determining the aspect ratio based on the surface density of the surface components and the surface density of the surface receptor. The method of claim 1, wherein the particle surface comprises a convex surface having local surface density and curvature of components effective for a preselected inclusion mode. A method of treating or monitoring a physiological condition comprising administering to a subject in need thereof the effective amount of the particle composition prepared according to the method of claim 12. A composition prepared according to the method of claim 1.

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