CN101652127A - Particles for cell targeting - Google Patents

Abstract

Compositions comprising oblate spheroidal particles comprising an active agent, such as a therapeutic or imaging agent, and methods of treating or monitoring a physiological condition (e.g., a disease) by administering the compositions to a subject in need thereof are provided. Also provided are methods for making particles having a volume capable of enhancing the attachment of the particles to a target site within a subject's body for a pre-selected shape of the particles, and methods for making particles having a shape capable of enhancing the attachment of the particles to a target site within a subject's body for a pre-selected volume of the particles.

Description

Particles for Cell Targeting

Statement Regarding Federally Funded Research or Development

not applicable.

field of invention

The present invention relates generally to targeted delivery of therapeutic and/or imaging agents, and more particularly to microparticles or nanoparticles, methods of producing such particles, and the use of such particles for targeting of therapeutic and/or imaging agents. to the method of delivery.

Background of the invention

Microparticles or nanoparticles with different compositions and chemical-physical properties can be used to deliver active agents, such as therapeutic or imaging agents, see e.g. LaVan D.A., et al. Small-scale systems for in vivo drug delivery. Nat. Biotechnol. 2003 21:1184-91; and Ferrari M..Curr.Opin.Chem.Biol.2005;9:343-6. Examples of such microparticles or nanoparticles include nanospheres, where loads, such as drug molecules or imaging agents, are dispersed in a polymeric matrix, see e.g. Duncan R. Nat. Rev. Drug Discov. 2003;2:347-60 ; multilamellar nano/microcapsules and liposomes, where the load is contained within the inner capsule, see e.g. Crommelin D.J.A., Schreier H., Liposomes, pp.73-190, in: Colloidal drug delivery systems, Kreuter J., editor, New York: Marcel Dekker, 1994; and nanoporous silicon particles, where the load is bound to a porous surface, see eg Cohen M.H., et al. Biomed. Microdev. 2003;5:253-9.

One of the advantages of microparticle or nanoparticle administration over free molecule administration is their versatility and engineerability. For example, microparticles or nanoparticles can carry high loads of therapeutic agents that can be released in precise doses and schedules, improving the efficacy and specificity of treatments. Microparticles or nanoparticles can carry both therapeutic and imaging agents so that the latter can monitor in vivo the development of a disease or physiological condition (eg a cancerous tumor) following therapeutic treatment. The surface of a microparticle or nanoparticle may have targeting moieties, such as different types of ligands capable of increasing the probability of specific recognition of the particle through the targeting site.

In order to perform their diagnostic and/or therapeutic tasks, the microparticles or nanoparticles must be tightly attached to one or more cells at the target site, eg damaged cells. This tight attachment can be particularly important for targeting vasculature sites, in which case the attachment interactions must counteract the hemodynamic forces exerted on the particles by blood flow that tends to expel the particles from the target site, see e.g. Neri D., Bicknell R. Nat. Cancer Rev. 2005. Therefore, there is a need to develop microparticles or nanoparticles with enhanced attachment to target sites.

Summary of the invention

One embodiment of the invention provides a method of treating or monitoring a physiological condition comprising administering to a subject in need thereof a composition comprising oblate spheroidal particles comprising an effective amount of at least one active agent.

Another embodiment of the present invention provides a composition comprising oblate spheroidal particles comprising at least one active agent.

In another embodiment, methods are provided comprising (A) selecting a target site having a surface having one or more first moieties; (B) selecting a second moiety complementary to the first moiety; ( C) selecting a shape defined by one or more shape parameters; (D) determining a volume that maximizes attachment to a target site, based on (i) the selected one or more shape parameters; (ii) the first part and one or more parameters of the interaction between the second moiety; and (iii) the surface density of the first moiety at the target site; and (E) making particles having a substantially selected shape and a substantially defined and (F) placing the second portion on the surface of the particle.

And in another embodiment, there is provided a method comprising (A) selecting a target site having a surface having one or more first moieties; (B) selecting a volume; (C) selecting a target site with the The second part complementary to the first part; (D) determining the volume that maximizes attachment to the target site, based on (i) the selected volume; (ii) the parameters of the interaction between the first part and the second part; and ( iii) surface density of the first moiety on the surface of the target site; and (E) fabricating a particle having a substantially defined shape and a substantially selected volume; and (F) placing the second moiety on the surface of the particle.

Description of drawings

Figure 1 illustrates a spherical particle attached to an endothelial substrate via ligand-receptor bonding.

作为体积V对球形粒子的长宽比γ的几种预选的数值(＝1、3、5、7和9)的函数的曲线图，其中m_r＝10¹⁴m^-2；μS＝1Pa；λ＝10^-10m；h₀＝10^-8m；δ_eq＝5×10^-9m。在

中，对应于最大的体积值是对于γ的具体预选值的最大体积V_opt。Figure 2 presents the dimensionless adhesion probability (dimensionless adhesion probability)

Graph as a function of the volume V for several preselected values (=1, 3, 5, 7 and 9) of the aspect ratio γ of spherical particles, where m _r =10 ¹⁴ m ⁻² ; μS=1 Pa; λ =10 ^-10 m; h ₀ =10 ^-8 m; δ _eq =5×10 ^-9 m. exist

, the corresponding maximum volume value is the maximum volume V _opt for a specific preselected value of γ.

中，对应于最大的长宽比是对于V的具体预选值的最大长宽比γ_opt。Figure 3 presents the dimensionless attachment probability As a graph of the aspect ratio γ of spherical particles as a function of several preselected values of the volume V varying in steps of 0.1 μm in ^{the range 0.1-1 μm 3 ,} Where μS = 0.5Pa; λ = 10 ^-10 m; h ₀ = 10 ^-8 m. exist

Among them, the maximum aspect ratio corresponding to the maximum aspect ratio is the maximum aspect ratio γ _opt for a specific preselected value of V.

Detailed description of the invention

definition

Unless stated otherwise, "a" or "an" means one or more.

"Microparticles" refers to particles having a largest characteristic dimension of 1 micron to 1000 microns, or in some embodiments, the range is 1 micron to 100 microns as specifically stated.

"Nanoparticles" refers to particles having a largest characteristic dimension of less than 1 micron.

By "oblate-spherical particle" is meant a particle having a substantially spherical shape with an aspect ratio [gamma] greater than 1. For the definition of the aspect ratio γ, refer to the following.

"Biodegradable" refers to a material capable of dissolving or degrading in a physiological matrix, or a biocompatible polymeric material capable of being degraded by physiological enzymes and/or under chemical conditions under physiological conditions.

review

The following research articles and patent documents, which are incorporated herein by reference in their entirety, are helpful in understanding the disclosure of the present invention:

1) P. Decuzzi and M. Ferrari. The adhesive strength of non-spherical particles mediated by specific interactions, Biomaterials 27(2006) 5307-5314;

2) P. Decuzzi et al. A Theoretical Model for the Margination of Particles within Blood Vessels, Annals of Biomedical Engineering 33(2005) 179-190;

3) P. Decuzzi et al. The Effective Dispersion of Nanovectors Within the Tumor Microvasculature, Annals of Biomedical Engineering 34(2006) 633-641;

4) P. Decuzzi et al. The Adhesion of Microfabricated Particles on Vascular Endothelium: Parametric Analysis, Annals of Biomedical Engineering 32(2004) 793-802;

5) U.S. Patent Application No. 11/836,004, filed August 8, 2007, pertaining to Ferrari;

6) PCT Application No. PCT/US2006/03986, filed September 27, 2006, by Decuzzi and Ferrari.

The inventors have realized that particles with an oblate spheroid are able to attach more strongly to endothelial cells than spherical particles. Accordingly, embodiments of the present invention provide compositions comprising oblate spheroidal particles comprising an active agent, such as a therapeutic or imaging agent, and by administering such a composition to a subject (e.g., a mammal, A method for treating or monitoring a physiological condition, such as a disease, preferably a human. Administration of oblate particles can reduce the effective amount of active agent used to treat or monitor a physiological condition compared to administration of particles having other shapes, such as spherical particles. Although the composition may also contain other particles that do not have an oblate shape, preferably, the oblate particles constitute at least 20%, or at least 30%, or at least 40%, at least 50%, or at least 60% of the total number of particles in the composition. %, or at least 70%, or at least 80%, or at least 90%. In some embodiments, substantially all of the particles in the composition are oblate spheroidal particles.

In some embodiments, the average aspect ratio of the oblate spheroidal particles is substantially equal to the attachment enhancement or maximum aspect ratio γ _opt for the average volume of the oblate spheroidal particles. The determination of the attached maximum aspect ratio _γopt for a given volume of oblate spheroidal particles is discussed below.

At the same time, the average aspect ratio of the oblate spheroidal particles may be such that the largest characteristic dimension a of the particle, which is half the length of the longer axis of the spheroid, is substantially smaller than a capillary at the body site targeted by the composition The average diameter r. Preferably, the largest characteristic dimension of the particles is at least 2 times or at least 4 times smaller than the mean capillary diameter at the targeted body site. The volume V, maximum characteristic size, and aspect ratio of spherical particles are related according to the following equation: $\mathrm{\γ}=\frac{{4\mathrm{\πa}}^3}{3V}.$ From this equation, _γmax that satisfies the above relationship between the largest characteristic dimension of the particle and the average diameter of the capillary at the targeted body site can be readily determined. Particles having an average aspect ratio substantially equal to γ _max may be used when γ _max is less than the average volume γ _opt of the particles.

A physiological condition that can be monitored or treated by oblate particles can be any condition that requires targeted delivery. For example, the physiological condition can be a disease, such as cancer or inflammation.

The present inventors have also discovered that microparticles or nanoparticles having a particular shape have a volume that can enhance or maximize the attachment of the particle to a particular target site. Meanwhile, the inventors have found that microparticles or nanoparticles having a specific volume have a volume that can enhance or maximize the attachment of the particle to a specific target site.

Accordingly, embodiments of the invention provide methods of making or designing microparticles or nanoparticles capable of enhanced attachment of cells to target sites. According to one embodiment, it is possible to (A) select a shape defined by one or more shape parameters, (B) select a target site having a surface with one or more first moieties thereon; The complementary second part, (D) determines the volume that maximizes attachment to the target site based on (i) the chosen shape, (ii) the parameters of the interaction between the first part and the second part, and (iii) the the surface density of a portion on the surface of the target site; and (E) fabricating a particle having a substantially selected shape and a substantially defined volume; and subsequently (E) placing a second portion on the surface of the particle. According to other embodiments, it is possible to (A) select a volume; (B) select a target site having a surface on which one or more first moieties are located; (C) select a second moiety complementary to the first moiety; (D) The shape that maximizes attachment to the target site is determined by one or more shape parameters based on (i) the selected volume, (ii) the parameters of the interaction of the first part and the second part, and (iii) the first part in the surface density on the surface of the target site; (E) fabricating a particle having a substantially defined shape and substantially selected volume; and subsequently (E) placing the second moiety on the surface of the particle. The particular volume of the particle can be selected based on the target loading of the active agent desired to be delivered to the target site.

In many embodiments, the selected target site is a vascular site, such as a coopted blood vessel, a new blood vessel, or a renormalized blood vessel, and the first portion is a molecular receptors. For example, for co-selected blood vessels, the first part is angiopoietin 2 (angiopoietin 2); for new blood vessels, the first part is vascular endothelial growth factor (VEGF), basic fibroblast growth factor or endothelial markers (such as α _v β ₃ integrins); for renormalized blood vessels, the first part is carcinoembryonic antigen-associated cell adhesion molecule 1 (CEACAM1), endothelin-B receptor (ET-B), vascular endothelial growth factor inhibitor gravin/AKAP12, protein Scaffolding protein for kinase A and protein kinase C.

The surface density on the first part can be determined using methods known to those skilled in the art. For example, when the first moiety is a molecular receptor, their surface density can be determined in vivo using a radiolabeled monoclonal antibody, as for Panes J., et al.Am.J.Physiol.1995; 269(6Pt2): Receptor complementary to the intracellular adhesion molecule 1 receptor discussed in H1955-64. Alternatively, surface density can be determined using fluorescently labeled monoclonal antibodies that are complementary to the receptor. Such fluorescently labeled monoclonal antibodies are, for example, antibodies labeled with phycoerythrin as disclosed in US Pat. No. 4,520,110.

The second moiety may be chosen to be complementary to the first moiety, ie the second moiety is capable of binding to the first moiety. For example, for a molecular receptor on a target vascular site, the second moiety is an antibody, aptamer or ligand capable of binding to the receptor.

The maximum adhesion force of the particle to the target site corresponds to the maximum dimensionless attachment probability ${\tilde{P}}_a=A_C\exp [-\frac{\mathrm{\λf}}{k_{B}T}],$ where _AC is the area of interaction between the microparticle or nanoparticle and the target site; λ is the characteristic length of the bond between the first part and the second part, such as a ligand-receptor bond, and f is each The force of a first part/second part pair, such as a ligand-receptor pair; k _B is the Boltzmann constant; T is the absolute temperature of the target site in Kelvins. Thus, the attached largest volume is the volume with the largest preselected shape for p˜a; and the attached largest shape is the shape with the largest preselected volume for P _a .

The following disclosure exemplifies the determination of the maximum volume of attachment and the maximum shape of attachment of spherical microparticles or nanoparticles, however it should be understood that similar methods can be used for non-spherical particles as well.

spherical particles

Figure 1 illustrates spherical particles with ligand surface density _m1 attached to a target site, which is an endothelial substrate with surface density _mr of receptor molecules.

For such spherical particles, choosing one or more shape parameters of the particle means choosing a specific aspect ratio γ = a/b, where a and b are half the lengths of the 2 different axes of the spherical particle, as in Cartesian described in child coordinates $\frac{x^2+{\mathrm{the\; y}}^2}{a^2}+\frac{z^2}{b^2}=1,$ where z is the axis of rotational symmetry. The volume of a spherical particle is related to the aspect ratio as follows: $V=\frac{4}{3}{\mathrm{\πa}}^3{\mathrm{\γ}}^{-1}.$

The area A _C of the interaction for spherical particles can be estimated as πr ₀ ² , where r ₀ is the diameter of the annular portion of the spherical particle at a separation distance h ₀ from the surface of the target site, where h ₀ is the maximum A distance at which specific bonding can still occur between a first moiety such as one or more molecular receptors and a second moiety such as one or more ligands. Estimate πr ₀ ² as follows:

$\mathrm{<span\; class="notranslate">\; \&pi;</span>}{\mathrm{<span\; class="notranslate">\; r</span>}}_{\mathrm{<span\; class="notranslate">\; 0</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; \&pi;a</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}<span\; class="notranslate">\; [</span>\mathrm{<span\; class="notranslate">\; 1</span>}<span\; class="notranslate">\; -</span>{<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; 1</span>}<span\; class="notranslate">\; -</span>\frac{{\mathrm{<span\; class="notranslate">\; h</span>}}_{\mathrm{<span\; class="notranslate">\; 0</span>}}<span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; \&delta;</span>}}_{\mathrm{<span\; class="notranslate">\; eq</span>}}}{\mathrm{<span\; class="notranslate">\; a</span>}}\mathrm{<span\; class="notranslate">\; \&gamma;</span>}<span\; class="notranslate">\; )</span>}^{\mathrm{<span\; class="notranslate">\; 2</span>}}<span\; class="notranslate">\; ]</span><span\; class="notranslate">\; ,</span>$

where δ _eq is the separation distance between the microparticle or nanoparticle and the surface of the target site (eg endothelial substrate). Figure 1 illustrates the parameters _AC , r ₀ , δ _eq and h ₀ .

The force f per unit of ligand-receptor binding can be expressed as the ratio between the total dislodging force F _dis and the interaction area A _C multiplied by the surface density m _r of the first moiety (e.g. molecular receptor), i.e. f=F _dis /(m _{r AC} ).

The total removal force F _dis consists of 2 parts: one part relates to the drag force F along the direction of flow in the blood vessel containing the target site, while the other part relates to the torque T exerted by the blood flow on the particles, see Fig. 1 . For spherical particles, the total removal force _Fdis can be described as follows:

F _dis ＝F+2T/r ₀ ＝6πa(aγ ^-1 +δ _eq )μSF ^S +8πa ³ μST ^S /r ₀ ,

where μ is the dynamic blood viscosity, and S is the blood shear rate, F ^S and T ^S are coefficients, which can be interpolated in Pozrikidis C. The motion of particles in the Hele-Shaw cell. J. Fluid . Mech. 1994; 261: 199-222 to estimate spherical and other non-spherical particles, which is hereby incorporated by reference in its entirety. Therefore, for spherical particles, F ^S and T ^S are described as follows:

F ^S ＝1+(1.736-0.138γ+0.128γ ² +0.09γ ³ )e ^−γ ;

T ^S =1+(-20.50+46.50γ-35.10γ ² +8.95γ ³ )e ^−γ .

For spherical particles, the dimensionless attachment probability is described as follows:

${\stackrel{<span\; class="notranslate">\; ~</span>}{\mathrm{<span\; class="notranslate">\; P</span>}}}_{\mathrm{<span\; class="notranslate">\; a</span>}}<span\; class="notranslate">\; =</span>{\mathrm{<span\; class="notranslate">\; \&pi;r</span>}}_{\mathrm{<span\; class="notranslate">\; 0</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}\mathrm{<span\; class="notranslate">\; exp</span>}<span\; class="notranslate">\; [</span><span\; class="notranslate">\; -</span>\frac{\mathrm{<span\; class="notranslate">\; \&lambda;</span>}}{{\mathrm{<span\; class="notranslate">\; k</span>}}_{\mathrm{<span\; class="notranslate">\; B</span>}}\mathrm{<span\; class="notranslate">\; T</span>}}<span\; class="notranslate">\; [</span>\mathrm{<span\; class="notranslate">\; 6</span>}<span\; class="notranslate">\; (</span>{\mathrm{<span\; class="notranslate">\; a\&gamma;</span>}}^{<span\; class="notranslate">\; -</span>\mathrm{<span\; class="notranslate">\; 1</span>}}<span\; class="notranslate">\; +</span>{\mathrm{<span\; class="notranslate">\; \&delta;</span>}}_{\mathrm{<span\; class="notranslate">\; eq</span>}}<span\; class="notranslate">\; )</span>{\mathrm{<span\; class="notranslate">\; f</span>}}^{\mathrm{<span\; class="notranslate">\; S</span>}}<span\; class="notranslate">\; +</span>\mathrm{<span\; class="notranslate">\; 8</span>}\frac{{\mathrm{<span\; class="notranslate">\; a</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}}{{\mathrm{<span\; class="notranslate">\; r</span>}}_{\mathrm{<span\; class="notranslate">\; 0</span>}}}{\mathrm{<span\; class="notranslate">\; T</span>}}^{\mathrm{<span\; class="notranslate">\; S</span>}}<span\; class="notranslate">\; ]</span>\frac{\mathrm{<span\; class="notranslate">\; a</span>}}{{\mathrm{<span\; class="notranslate">\; r</span>}}_{\mathrm{<span\; class="notranslate">\; 0</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}}\frac{\mathrm{<span\; class="notranslate">\; \&mu;S</span>}}{{\mathrm{<span\; class="notranslate">\; m</span>}}_{\mathrm{<span\; class="notranslate">\; r</span>}}}<span\; class="notranslate">\; ]</span><span\; class="notranslate">\; .</span>$

求导，并且使用例如数字或图形方法，设定关于a的

的一阶导数等于0，求a_opt。体积V_opt如下与a_opt相关联：To determine the maximum volume of attachment V _opt for a preselected γ, with respect to a pair

take the derivative, and using e.g. numerical or graphical methods, set with respect to a

The first derivative of is equal to 0, find a _opt . The volume V _opt is related to a _opt as follows:

${\mathrm{<span\; class="notranslate">\; V</span>}}_{\mathrm{<span\; class="notranslate">\; opt</span>}}<span\; class="notranslate">\; =</span>\frac{\mathrm{<span\; class="notranslate">\; 4</span>}}{\mathrm{<span\; class="notranslate">\; 3</span>}}\mathrm{<span\; class="notranslate">\; \&pi;</span>}{\mathrm{<span\; class="notranslate">\; a</span>}}_{\mathrm{<span\; class="notranslate">\; <figure-callout\; id="3"\; label="opt"\; filenames="A2007800380550033H1.png"\; state="\{\{state\}\}">opt</figure-callout></span>}}^{\mathrm{<span\; class="notranslate">\; 3</span>}}{\mathrm{<span\; class="notranslate">\; \&gamma;</span>}}^{<span\; class="notranslate">\; -</span>\mathrm{<span\; class="notranslate">\; 1</span>}}<span\; class="notranslate">\; .</span>$

求导，并且使用例如数字或图形方法，设定关于γ的的一阶导数等于0，求γ_opt。Similarly, to determine the maximum attachment parameter γ _opt , for γ pair

take the derivative, and using e.g. numerical or graphical methods, set with respect to γ The first derivative of is equal to 0, find γ _opt .

对于球形粒子的长宽比γ(＝1、3、5、7和9)的几个预选值的图形，其中m_r＝10¹⁴m²；μS＝1Pa；λ＝10^-10m；h₀＝10^-8m；δ_eq＝5×10^-9m。对应于最大

的体积值是对于γ特定的预选值的附着最大体积V_opt。Figure 2 shows the dimensionless attachment probability as a function of volume V

Graph for several preselected values of aspect ratio γ (=1, 3, 5, 7 and 9) for spherical particles, where m _r =10 ¹⁴ m ² ; μS=1 Pa; λ=10 ⁻¹⁰ m; h ₀ =10 ^-8 m; δ _eq =5×10 ^-9 m. corresponds to the maximum

The volume value for is the attached maximum volume V _opt for a specific preselected value of γ.

的长宽比值是对于V特定的预选值的附着最大长宽比γ_opt。Figure 3 shows the dimensionless attachment probability as a function of the aspect ratio γ of spherical particles Graph for several preselected values of volume V (range 0.1-1 μm ³ , step size 0.1 μm ³ ) with S=0.5 Pa; λ=10 ⁻¹⁰ m; h ₀ =10 ⁻⁸ m. corresponds to the maximum

The aspect ratio value of is the attached maximum aspect ratio γ _opt for a specific preselected value of V .

表达式中的所有参数基于所选的靶位点以及第一部分和第二部分之间的相互作用的特性和参数。Before making particles, determine the value of V _opt or γ _opt , because

All parameters in the expression are based on the chosen target site and the properties and parameters of the interaction between the first and second moieties.

For example, for blood viscosity μ, an average value of 10 ^-3 Pa s can be used for humans, or alternatively determined experimentally from plasma viscosity, hematocrit and mean wall shear rate measured using a glass capillary viscometer Values of blood viscosity, as disclosed in Weaver JPet al.Clin.Sci.36:1-10, 1969 and Dammers R., et al.J.Appl.Physiol.94:485-489, 2003, both by reference The entirety of which is incorporated herein, while the blood shear rate S can be assessed non-invasively in vivo using an ultrasound system as described by Dammers R., et al. J. Appl. Physiol. 94:485-489, 2003. Table 1 provides typical values of blood share rate for selected blood vessels in humans

Table 1

Blood vessel µS, Pa aorta 2.5 arteries 5 Arterioles 7.5 Capillaries 10 venules 0.2 vein 0.5 large vein 1

h ₀ , the maximum distance at which specific bonding between a first moiety (e.g. a molecular receptor) and a second moiety (e.g. a ligand) can still occur and is determined by, for example, a change in the length of the linker moiety of the second moiety control.

λ, the characteristic length of the bond between the first moiety and the second moiety, which depends on the first moiety and the chosen second moiety on the target surface. For example, when the first part is a molecular receptor and the second part is a ligand, λ as in Dembo, M., DC Torney, K. Saxaman, and D. Hammer. 1988. The reaction-limited kinetics of membrane-to-surface adhesion and detachment .Proc.R.Soc.Lond.B.234:55-83, which is hereby incorporated by reference in its entirety. For a typical receptor-ligand pair, λ is about 1

δ _eq , the separation distance between the microparticle or nanoparticle and the surface of the target site (e.g. the endothelial substrate in FIG. 1 ), which can be obtained by solving the following equation for δ using, for example, numerical or graphical methods:

$\frac{{\mathrm{<span\; class="notranslate">\; A</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}}{{\mathrm{<span\; class="notranslate">\; <figure-callout\; id="12"\; label="k\; B\; T"\; filenames="A2007800380550034H1.png"\; state="\{\{state\}\}">k</figure-callout></span><figure-callout\; id="12"\; label="k\; B\; T"\; filenames="A2007800380550034H1.png"\; state="\{\{state\}\}">\; </figure-callout>}}_{\mathrm{<span\; class="notranslate">\; B</span>}}\mathrm{<span\; class="notranslate">\; T</span>}\mathrm{<span\; class="notranslate">\; 12</span>}\mathrm{<span\; class="notranslate">\; \&pi;\&delta;</span>}}<span\; class="notranslate">\; +</span>\frac{\mathrm{<span\; class="notranslate">\; 64</span>}{\mathrm{<span\; class="notranslate">\; \&rho;</span>}}_{<span\; class="notranslate">\; \&infin;</span>}{\mathrm{<span\; class="notranslate">\; a</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}}{\mathrm{<span\; class="notranslate">\; \&kappa;</span>}}{\mathrm{<span\; class="notranslate">\; z</span>}}_{\mathrm{<span\; class="notranslate">\; v</span>}}{\mathrm{<span\; class="notranslate">\; z</span>}}_{\mathrm{<span\; class="notranslate">\; c</span>}}{\mathrm{<span\; class="notranslate">\; e</span>}}^{<span\; class="notranslate">\; -</span>\mathrm{<span\; class="notranslate">\; \&kappa;\&delta;</span>}}<span\; class="notranslate">\; -</span>\mathrm{<span\; class="notranslate">\; 36</span>}\mathrm{<span\; class="notranslate">\; \&Gamma;</span>}{\mathrm{<span\; class="notranslate">\; a</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}{\mathrm{<span\; class="notranslate">\; e</span>}}^{<span\; class="notranslate">\; -</span>\mathrm{<span\; class="notranslate">\; \&delta;</span>}<span\; class="notranslate">\; /</span>{\mathrm{<span\; class="notranslate">\; R</span>}}_{\mathrm{<span\; class="notranslate">\; g</span>}}}<span\; class="notranslate">\; =</span>\mathrm{<span\; class="notranslate">\; 0</span>}<span\; class="notranslate">\; .</span>$

In the above equation, A is the Hamacker constant, which is estimated using the following formula:

$\mathrm{<span\; class="notranslate">\; A</span>}<span\; class="notranslate">\; \&ap;</span>\frac{\mathrm{<span\; class="notranslate">\; 3</span>}}{\mathrm{<span\; class="notranslate">\; 4</span>}}{\mathrm{<span\; class="notranslate">\; k</span>}}_{\mathrm{<span\; class="notranslate">\; B</span>}}\mathrm{<span\; class="notranslate">\; T</span>}<span\; class="notranslate">\; (</span>\frac{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 1</span>}}<span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 3</span>}}}{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 1</span>}}<span\; class="notranslate">\; +</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 3</span>}}}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; (</span>\frac{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 2</span>}}<span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 3</span>}}}{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 2</span>}}<span\; class="notranslate">\; +</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 3</span>}}}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; +</span>\frac{\mathrm{<span\; class="notranslate">\; 3</span>}\mathrm{<span\; class="notranslate">\; h</span>}}{\mathrm{<span\; class="notranslate">\; 4</span>}\mathrm{<span\; class="notranslate">\; \&pi;</span>}}\underset{{\mathrm{<span\; class="notranslate">\; <figure-callout\; id="1"\; label="v"\; filenames="A2007800380550033H1.png,A2007800380550034H1.png"\; state="\{\{state\}\}">v</figure-callout></span>}}_{\mathrm{<span\; class="notranslate">\; 1</span>}}}{\overset{<span\; class="notranslate">\; \&infin;</span>}{<span\; class="notranslate">\; \&Integral;</span>}}<span\; class="notranslate">\; (</span>\frac{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 1</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; iv</span>}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 3</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; iv</span>}<span\; class="notranslate">\; )</span>}{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 1</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; iv</span>}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; +</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 3</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; iv</span>}<span\; class="notranslate">\; )</span>}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; (</span>\frac{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 2</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; iv</span>}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; -</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 3</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; iv</span>}<span\; class="notranslate">\; )</span>}{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 2</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; iv</span>}<span\; class="notranslate">\; )</span><span\; class="notranslate">\; +</span>{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 3</span>}}<span\; class="notranslate">\; (</span>\mathrm{<span\; class="notranslate">\; iv</span>}<span\; class="notranslate">\; )</span>}<span\; class="notranslate">\; )</span>\mathrm{<span\; class="notranslate">\; dv</span>}<span\; class="notranslate">\; ,</span>$

where ε ₁ , ε ₂ and ε ₃ are the static (DC) dielectric constants of particles, endothelial cells and liquid components of blood (plasma), respectively; , ε ₁ (iv), ε ₂ (iv) and ε ₃ (iv) are the numerical dielectric functions of particles, endothelial cells and liquid components of blood (plasma), respectively; v ₁ =2πk _BT /h, h is Planck's constant. Dielectric functions and constants can be estimated using dielectric spectroscopy as published by C. Prodan, F. Mayo, JR Laycomb, and JH Miller, Jr., MJ Benedik, Low-frequency, low-field dielectric spectroscopy of living cell suspensions, Journal of AppliedPhysics -April 1, 2004-Volume 95, Issue 7, pp.3754-3756, the contents of which are hereby incorporated by reference in their entirety. Typical values for the Hamaker constant in liquids are about ^10-20 Joules, see eg Israelachvili, J. 1992, Intermolecular and Surface Forces, 2nd ed. Academic Press, New York.

ρ _∞ is the ion concentration of blood. Typical values for particle concentrations in blood are around 150 mM, see eg Ganong, WF Review of Medical Physiology, 21st ed. New York: Lange Medical Books/McGraw-Hill Medical Publishing Division, 2003.

κ ^-1 is the Debye length, the length above the electric field that a mobile charge carrier (eg electron) can scan out. Typically, in electrolytes such as blood, the Debye length is determined using the following formula:

${\mathrm{<span\; class="notranslate">\; \&kappa;</span>}}^{<span\; class="notranslate">\; -</span>\mathrm{<span\; class="notranslate">\; 1</span>}}<span\; class="notranslate">\; =</span>\sqrt{\frac{{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; 0</span>}}{\mathrm{<span\; class="notranslate">\; \&epsiv;</span>}}_{\mathrm{<span\; class="notranslate">\; r</span>}}{\mathrm{<span\; class="notranslate">\; k</span>}}_{\mathrm{<span\; class="notranslate">\; B</span>}}\mathrm{<span\; class="notranslate">\; T</span>}}{\mathrm{<span\; class="notranslate">\; 2</span>}{\mathrm{<span\; class="notranslate">\; N</span>}}_{\mathrm{<span\; class="notranslate">\; A</span>}}{\mathrm{<span\; class="notranslate">\; e</span>}}^{\mathrm{<span\; class="notranslate">\; 2</span>}}\mathrm{<span\; class="notranslate">\; I</span>}}}<span\; class="notranslate">\; ,</span>$

where _ε0 is the vacuum permittivity, _εr is the dielectric constant of the electrolyte, _kB is the Boltzmann constant, T is the absolute temperature, e is the charge on the electron, I is the particle strength of the electrolyte, and N _A is the avolt Gadlow's constant. For blood, the Debye length is about 0.8 nm.

Γ is the number of polymer chains per unit area. Γ = s ^-2 , where s is the average separation distance s between 2 adjacent chains on the nanoparticle surface. The separation distance s depends on the size of the functional groups on the nanoparticle surface and the size (molecular weight) of the polymer chains conjugated to the functional groups. The separation distance s can be estimated by citofluorometric exams, see eg Jacob N. Israelachvili, Intermolecular and Surface Forces, Second Edition: With Applications to Colloidal and Biological Systems, Academic Press; II Edition, 1992.

_Rg is the diameter of gyration of the polymer (eg, ligand). R _g is related to the number N of polymer repeat units forming the polymer chain and the effective length / of the repeat units. Also depends on the solvent of the polymer. For an ideal solution, that is, one in which the interactions (attraction-repulsion) between repeating units of the polymer are negligible, $R_g=l\sqrt{\frac{N}{6}}..$ For "good" solvents, ie solvents with repulsive forces between fragments, R _g = 1N ^3/5 ; for "bad" solvents, ie solvents with attractive interactions between repeating units, R _g = 1N ^{1 /3} See, eg, Jacob N. Israelachvili, Intermolecular and Surface Force,: With Applications to Colloidal and Biological Systems, Academic Press; Second Edition, 1992. The liquid component of blood (plasma) is a liquid solution, and water is a good solvent for PEG polymers.

z _v and z _c are the electrostatic surface potentials on the particle surface and on the target surface, respectively. [epsilon _]v and [epsilon _]c can be estimated using the Zetasizer ^™ Nano series equipment from Malvern Instruments, Worcestershire United Kingdom.

make

After determining the maximum volume of attachment for the preselected shape, particles having a volume substantially of the maximum volume of attachment and having a shape substantially determined by one or more preselected shape parameters can be produced. Similarly, after determining the maximum shape parameter of attachment for a preselected volume, particles can be produced having a volume substantially of the preselected volume and a shape substantially determined by the maximum shape parameter of attachment. Then use the second part to modify the produced particles.

With respect to volume, the term "substantially" means that the volume is as close to a preselected or determined volume as the particular manufacturing method allows. The produced volume is thus within ±30% or ±20% or ±10% or ±5% or ±3% of the preselected or determined volume.

With respect to shape, the term "substantially" means that the shape is as close to a preselected or determined shape as the particular fabrication method allows. For example for spherical particles, the aspect ratio is fabricated within ±30% or ±20% or ±10% or ±5% or ±3% or ±1% of a preselected or defined volume.

The particles can be made by any of a variety of methods. In some embodiments, the particles are fabricated as detailed in van Dillen T., van Blaaderen A., Polman A. Ion beamshaping of colloidal assemblies. Mater. Today 2004:40-6, which is incorporated by reference in its entirety into this article. This technique can be used to convert spherical silica particles to oblate and ellipsoidal shapes.

In some embodiments, the particles are fabricated as bubbles or droplets that exist in a stable non-spherical shape, as in Subramaniam A.B., Abkarian M., Mahadevan L., Stone H.A. Nonspherical bubbles. Nature 2005;438:930 details, which are hereby incorporated by reference in their entirety.

In some embodiments, the particles are fabricated using the particle replication in non-wetting templates (PRINT) technique, such as in Rolland J.P., Maynor B.W., Euliss L.E., ExnerA.E., Denison G.M., DeSimone J. Direct fabrication and harvesting of monodisperse, shape specific nano-biomaterials. J. Am. Chem. Soc. 2005; 127: 10096-100, which is incorporated herein by reference in its entirety. This technique is very versatile and flexible, and enables the fabrication of particles with simultaneous control over shape, size, composition, cargo and surface structure.

In some embodiments, the particles are fabricated by top-down microfabrication or nanofabrication methods, such as photolithography, e-beam lithography, X-ray lithography, deep ultraviolet lithography, or nanoimprint photolithography. prepared particles. A potential advantage of using such top-down fabrication methods is that these methods provide scaled-up production of uniformly sized particles.

After fabrication, a second moiety, such as a ligand, can be placed on the surface of the particle. For example, ligands may be chemically attached to suitable reactive groups on the surface of the particle. Protein ligands can be attached to amino- or thiol-reactive groups under conditions effective to form thioether or amide bonds, respectively. Methods for linking antibodies or other polymeric binding reagents to inorganic or polymeric supports are as detailed in Taylor, R., Ed., Protein Immobilization Fundamentals and Applications, pp. 109110 (1991). Preferably, the second moiety is placed in such a way that the surface density of the second moiety is greater than the surface density of the first moiety at the target site.

In some embodiments, a particle is fabricated having a body as defined by the volume and shape of the particle and one or more reservoirs within the body into which one or more active agents can be loaded.

In some embodiments, the particle has one or more channels connecting the reservoir and the surface. In some embodiments, the reservoirs and channels are pores within the body of the primary particle. In this case, the primary particles contain porous or nanoporous material. Preferably, the pores of the porous or nanoporous material are controlled to obtain a desired loading of the next stage particles and a desired release rate. Nanoporous materials with controllable pore size are oxide materials such as silicon oxide, aluminum oxide, titanium oxide or iron oxide. The fabrication of nanoporous oxide particles (also known as sol-gel particles), in for example Paik J.A.et.al.J.Mater.Res., Vol.17, Aug 2002.The nanoporous material with controllable pore size may be also nanoporous silicon described in detail in , which is incorporated herein by reference in its entirety. The nanoporous material with controllable pore size can also be nanoporous silicon. For details on the fabrication of nanoporous silicon particles, refer to Cohen M.H. et.al. Biomedical Microdevices 5:3, 253-259, 2003.

In some embodiments, the primary particle has no channels at all. The particles contain, for example, biodegradable materials. For example, the particles are composed of metals such as iron, titanium, gold, silver, platinum, copper, and alloys and oxides thereof. The biodegradable material is also a biodegradable polymer such as polyorthoesters, polyanhydrides, polyamides, polyalkylcyanoacrylates, polyphosphazenes, and polyesters. Examples of biodegradable polymers are described in detail, eg, in U.S. Pat. Nos. 4,933,185, 4,888,176 and 5,010,167. Specific examples of such biodegradable polymers include poly(lactic acid), polyglycolic acid, polycaprolactone, polyhydroxybutyric acid, poly(N-palmitoyl-trans-4-hydroxy-L-proline esters) and poly(DTH carbonate).

In some embodiments, the fabricated particle is the active agent itself.

Loading active reagents

In some embodiments, the methods of the invention further comprise loading the particles with an active agent. The particular loading technique depends on the composition of the particles. For example, a particle fabricated from a nanoporous material is soaked in a solution containing a carrier fluid and an active agent, which enters the pores of the precursor particle by capillary action. The carrier fluid is a biologically harmless liquid that is neutral to the active agent. Examples of carrier fluids are phosphate buffered saline (PBS) or deionized water. To maximize the loading of the active agent, eg a solution with a saturating concentration of the active agent is used.

The solution containing the active agent was degassed prior to the introduction of the particles. The particles are then immersed in a degassed solution in a sealed chamber. The particles are placed under reduced pressure to ensure that entrapped air is expelled from the pores in the particles. The particles are then completely submerged in the solution and the pressure in the sealed chamber is raised to slightly above atmospheric pressure to ensure that the solution enters the pores of the particles. The particles were then captured with a filter and dried using one of the three methods described below.

To remove any air trapped in the reservoir of soaked particles, the pressure in the chamber was reduced and then raised to slightly above atmospheric pressure.

After pouring the solution into the pores of the particles, drying is achieved by one or more of the following three methods. Water is removed by evaporation under reduced pressure in a vacuum chamber, or by blowing a stream of warm air or an inert gas (eg nitrogen) over the surface particles collected on a strainer, or by freeze drying. In the case of freeze-drying, a flat heat exchanger is placed in a position of good thermal contact with the filter collecting the foreline particles, for example directly below it. Refrigerant liquid (such as freon), or cold liquid (such as liquid nitrogen) in the temperature range of -20°C to -60°C is passed through the inlet and outlet of the heat exchanger to freeze any water remaining in the pores. The pressure is then reduced until all the water sublimates.

active agent

The active agent is a therapeutic compound or an imaging moiety. The active agent is any suitable agent. In some embodiments, the active agent is fabricated into particles. In some embodiments, the active agent is an agent released from the particle into which it is incorporated. The choice of active agent depends on the application.

A therapeutic agent is any physiologically or pharmacologically active substance that produces a desired biological effect at a targeted site in a subject, such as a mammal or a human. A therapeutic agent is any inorganic or organic compound without limitation, including any identified or unidentified peptides, proteins, nucleic acids, and small molecules. Therapeutic agents exist in various forms such as unchanged molecules, molecular complexes, pharmaceutically acceptable salts such as hydrochloride, hydrobromide, sulfate, laurate, palmitate, phosphate, nitrous acid Salt, Nitrate, Borate, Acetate, Maleate, Tartrate, Oleate, Salicylate, etc. For acidic treatments, salts of metals, amines or organic cations, eg, quaternary ammoniums, are used. Derivatives of drugs such as bases, esters and amides are also used as therapeutic agents. Use of water-insoluble therapeutic agents in the form of their water-soluble derivatives, or their base derivatives, in either case, or delivered by them, converted by enzymes, hydrolyzed by body pH, or passed through Additional metabolic processes lead to the original therapeutically active form.

Therapeutic agents are chemotherapeutic agents, immunosuppressive agents, cytokines, cytotoxic agents, nucleolytic compounds, radioisotopes, receptors, and prodrug activating enzymes, or any combination thereof, either natural or produced synthetically or recombinantly.

Drugs affected by typical multidrug resistance, such as vinca alkaloids (eg, vinblastine and vincristine), anthracyclines (eg, doxorubicin and daunorubicin), RNA transcription inhibitors ( For example, actinomycin-D) and microtubule stabilizing drugs (eg, paclitaxel) have particular use as therapeutic agents.

Cancer chemotherapeutic agents are preferred therapeutic agents. Available cancer chemotherapy drugs include nitrogen mustards, nitrosoureas, ethyleneimines, alkyl sulfonates, tetrazines, platinum compounds, pyrimidine analogs, purine analogs, antimetabolites, folic acid analogs, anthracyclines , taxanes, vinca alkaloids, topoisomerase inhibitors and hormone reagents. Examples of chemotherapeutic drugs are Actinomycin-D, Acran, Cytarabine, Anastrozole, Asparaginase, BiCNU, Bicalutamide, Bleomycin, Busulfan, Capecitabine , Carboplatin, Carboplatinum, Carmustine, CCNU, Chlorambucil, Cisplatin, Cladribine, CPT-11, Cyclophosphamide, Cytarabine, Cytarabine (Cytosine arabinoside), cyclophosphamide, dacarbazine, actinomycin D, daunorubicin, dexrazoxane, docetaxel, doxorubicin, DTIC, epirubicin, ethyleneimine, Etoposide, floxuridine, fludarabine, fluorouracil, flutamide, formustine, gemcitabine, Herceptin, hexamethylammonium, hydroxyurea, idarubicin, ifosfamide, iritinib Kang, lomustine, nitrogen mustard, melphalan, mercaptopurine, methotrexate, mitomycin, mitotane, mitoxantrone, oxaliplatin, paclitaxel, pamidronic acid, pensi Statins, plicamycin, procarbazine, rituximab, steroids, streptozocin, STI-571, streptozocin, tamoxifen, temozolomide, teniposide, tetrazine, thioguanine , thiotepa, Tuoyoude, topotecan, trovasufan, trimethrexate, vinblastine, vincristine, vindesine, vinorelbine, VP-16 and Xeloda.

；雷佐生；西佐喃(Sizofrran)；锗螺胺；替奴佐酸；三亚胺醌；2，2’，2”-三氯三乙胺；乌拉坦；长春地辛；达卡巴嗪；甘露醇氮芥、二溴甘露醇；二溴卫矛醇；哌泊溴烷；Gacytosine；阿拉伯糖苷(“Ara-C”)；环磷酰胺；噻替哌；紫杉烷，例如紫杉醇(TAXOL

Bristol-Myers Squibb Oncology，Princeton，NJ)和多西紫杉醇(TAXOTERE

，Rhone-Poulenc Rorer，Antony，France)；苯丁酸氮芥；吉西他滨；6-硫鸟嘌呤；巯基嘌呤；甲氨喋呤；铂类似物例如顺铂和卡铂；长春碱；铂；依托泊苷(VP-16)；异环磷酰胺；丝裂霉素C；米托蒽醌；长春新碱；长春瑞滨；诺维本；Novantrone；替尼泊甙；柔红霉素；氨基喋呤；希罗达；伊班膦酸；CPT-11；拓扑异构酶抑试剂RFS 2000；二氟甲基鸟氨酸(DMFO)；视黄酸；Esperamicins、卡培他滨和以上任何一种的药学可接受的盐、酸或衍生物。也包括作用以调节或抑制激素对肿瘤的作用的抗激素试剂，例如抗雌激素药物包括例如他莫昔芬、雷洛西芬、芳香化酶抑制4(5)-咪唑、4羟基他莫昔芬、曲沃昔芬、雷洛西芬、奥那司酮和托瑞米芬(法乐通)；和抗雄激素药物例如氟他胺、尼鲁米特、比卡鲁胺、亮丙瑞林、和戈舍瑞林；和以上任何一种上述物质的药学可接受的盐、酸或衍生物。Useful cancer chemotherapeutic agents also include alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzotepa ( Benzodopa), Carboquinone, Meturedopa, and Uredopa; ethyleneimines and methylamelamines, including hexamethylmelamine, triethylenemelamine, triethylenemelamine Phosphoramides, triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards such as chlorambucil, naphthalene, cyclophosphamide, estramustine, ifosfamide, nitrogen mustards, Nitromustine hydrochloride, melphalan, nemethambucil, cholesteryl phenylacetate mustard, prednimustine, trofosamide, uracil mustard; nitroureas such as Cannustine, chlorurecin, formoxetine Nimustine, lomustine, nimustine, and ramustine; antibiotics such as aclarithromycin, actinomycin, Authramycin, azaserine, bleomycin, actinomycin C, calicheamicin Carabicin, Carabicin, Carbinycin, Carcinophilin, Chromoinycins, Actinomycin C, Daunorubicin, Detorubicin, 6-diazo-5-oxo-L-norleucine, poly Ruubicin, Epirubicin, Esorubicin, Idarubicin, Maxiclomycin, Mitomycin, Mycophenolic Acid, Nogamycin, Olivomycin, Pelomycin, Porphyra Potfiromycin, puromycin, triiron doxorubicin, rhodorubicin, streptomycin, streptozocin, tubercidin, ubenimex, netastatin, and zorubicin; antimetabolites drugs such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as methotrexate, pteroxate, and trimetrexate; purine analogs such as fludarabine, 6- Mercaptopurine, thiomethopurine, and thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azacitidine, carmofur, cytarabine, dideoxyuridine, doxifluridine , enoxitabine, floxuridine, and 5-FU; androgens such as caruterone, drotandrosterone propionate, ethiosterol, Rnepitiostane, and testolactone; antiadrenal agents such as aminoglutethimide, mitol Trilosteine; folic acid supplements such as folinic acid; acetglucuronolactone; aldophosphamide glycosides; aminolevulinic acid; amsacrine; Bestrabucil; bisantrene; edatrexate; Defofamine; colcemid ;Deacriquinone; Elfornithine; Etriacetium; Etoglu; Gallium nitrate, Hydroxyurea, Lentinan, Lonidamine, Mitoguanidine hydrazone, Mitoxantrone, Mopedadol, Nitrazine; pentostatin; methamine; pirarubicin; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK

; Razoxane; Sizofrran; Germanospiramine; Tenuzolic Acid; Triiminequinone; 2,2',2"-Trichlorotriethylamine;Urethane;Vindesine; Mechlorethamine, Dibromomannitol; Dibromodulcitol; Pipobromide; Gacytosine; Arabinoside ("Ara-C");Cyclophosphamide;Thiotepa; Taxanes such as Paclitaxel (TAXOL

Bristol-Myers Squibb Oncology, Princeton, NJ) and docetaxel (TAXOTERE

, Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etopol Glycoside (VP-16); Ifosfamide; Mitomycin C; Mitoxantrone; Vincristine; Vinorelbine; Navelbine; Novantrone; Teniposide; Daunorubicin; Amopterin ; Xeloda; Ibandronic acid; CPT-11; Topoisomerase inhibitor RFS 2000; Difluoromethylornithine (DMFO); Retinoic acid; Esperamicins, capecitabine, and any of the above Pharmaceutically acceptable salts, acids or derivatives. Also included are antihormonal agents that act to modulate or inhibit the effects of hormones on tumors, such as antiestrogens including, for example, tamoxifen, raloxifene, aromatase inhibitory 4(5)-imidazole, 4-hydroxytamoxifen fen, travoxifene, raloxifene, onapristone, and toremifene (Pharlotone); and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide Lin, and goserelin; and a pharmaceutically acceptable salt, acid or derivative of any of the above substances.

Cytokines are also used as therapeutic agents. Examples of such cytokines are lymphokines, monokines and traditional polypeptide hormones. Included among cytokines are growth hormones such as human growth hormone, N-methionyl human growth hormone, and bovine growth hormone; parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; Protein hormones such as follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), and luteinizing hormone (LH); hepatocyte growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor-alpha and -beta Mullerian Inhibitory Substance; Mouse Gonadotropin-Related Peptide; Inhibin; Activin; Vascular Endothelial Growth Factor; Integrin; Thrombopoietin (TPO); Nerve Growth Factors such as NGF-β; (TGF) such as TGF-α and TGF-β; insulin-like growth factors-I and -II; erythropoietin (EPO); osteogenic factors; interferons such as interferon-α, -β and -γ; colony stimulating Factors (CSF) such as macrophage-CSF (M-CSF); granulocyte-macrophage-CSF (GM-CSF); and granulocyte-CSF (GCSF); interleukins (IL) such as IL-1, IL-1a, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12, IL-15; tumor necrosis factors 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 of natural origin or from recombinant cell culture and biologically active equivalents of native sequence cytokines.

An imaging agent is any substance that provides imaging information about a targeted site in an animal (eg, a mammal or a human). Imaging agents include magnetic materials such as iron oxide for magnetic resonance imaging. For optical imaging, the active agents are, for example, semiconductor nanocrystals or quantum dots. For optical coherence tomography imaging, the imaging agent is a metal, eg gold or silver, nanocage particles. The imaging agent may also be an ultrasound contrast agent, such as microbubbles or nanobubbles or iron oxide microparticles or nanoparticles.

combination

Compositions comprising a plurality of such particles are also provided. Such a composition may be a suspension of the particles described above for administering a therapeutic or imaging agent to a subject. To form the suspension, the particles can be suspended in a liquid matrix at a concentration of choice. The optimal concentration will depend on the characteristics of the particles (eg dissolution properties), the type of therapeutic application and the mode of administration. For example, compositions for oral administration may be relatively viscous and thus may contain high concentrations (eg >50%) of the particles. Solutions for bolus injection preferably comprise a relatively concentrated suspension of the particles (e.g. 10-50%), but not so concentrated that it has a viscosity slightly higher than saline (to minimize the need for large bore needles) . Solutions for continuous intravenous infusion typically contain relatively low concentrations of particles (eg, 2-10% in suspension) because of the relatively large volumes of liquid to be administered.

The particles are suspended in any suitable liquid carrier. A suitable pharmaceutical carrier is one that is nontoxic to recipients at the dosages and concentrations employed and that is compatible with the other ingredients of the formulation. Examples of suitable carriers include, but are not limited to, water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Suspensions used in injectable formulations are preferably isotonic with the blood of the subject. Typically, the carrier contains minor amounts of additives, such as 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 dextran), chelating agents (such as EDTA), or other excipients.

The suspension of particles is sterilized by a suitable sterilization method prior to administration to a subject. Particles made from heat stable materials can be heat sterilized, for example using autoclaving. Particles made from thermally non-stable materials can be sterilized by passing through commercially available sterile filters, such as 0.2 μm filters. Of course, the filtration method can only be used when the particles are smaller than the pores of the sterile filter.

The particles are administered to a subject in need of therapeutic intervention by any suitable method of administration. The particular method for a particular application will be determined by the attending physician. The particles are administered by one of the following routes: topical, parenteral, inhalation, oral, vaginal and anal. Intravascular administration, including intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) injections, is particularly preferred.

Intravascular administration is local or systemic. Localized intravascular delivery is used to bring the particles into the vicinity of known lesions by using a catheter system, such as a CAT-scan catheter. Conventional injections such as bolus i.v. injections or continuous/trick i.v. infusions are generally systemic. Preferably, the particles are injected eg into the bloodstream and allowed to circulate and localize to their target sites.

While certain preferred embodiments have been indicated above, it should be understood that the invention is not limited thereto. Various modifications to the disclosed embodiments may occur to those skilled in the art and are intended to be within the scope of the invention.

All documents, patent applications and patents cited in this specification are hereby incorporated by reference in their entirety.

other references

1. LaVan DA, McQuire T, Langer R. Small-scale systems for in vivo drug delivery. Nat Biotechnol 2003; 21: 1184-91.

2.Ferrari M.Nanovector therapeutics.Curr Opin Chem Biol2005;9:343-6.3.Duncan R.The dawning era of polymer therapeutics.Nat Rev Drug Discov 2003;2:347-60.

4. Crommelin DJA, Schreier H, Liposomes. In: Kreuter J, editor. Colloidal drug delivery systems. New York: Marcel Dekker.

5. Cohen MH, Melnik K, Boiarski AA, Ferrari M, Martin FJ. Microfabrication of silicon-based nanoporous particulates formal applications. Biomed Microdev 2003;5:253-9.

6. Ferrari M. Cancer nanotechnology: opportunities and challenges. Nat Rev Cancer 2005;5:161-71.

7. Neri D, Bicknell R. Tumor vascular targeting. Nat Cancer Rev 2005. August. Vivek R, Patil S, Campbell CJ, Yun YH, Slack SM, Goetz DJ. Particle diameter influences adhesion under flow. Biophys J 2001; 80: 1733- .

9. Blackwell JE, Dagia NM, Dickerson JB, Berg EL, Goetz DJ. Ligand coated nanosphere adhesion to E-and P-selectin underwater and flow conditions. Ann Biomed Eng 2001; 29:523-33.10. Pierres A, Benoliel A-M, Zhu C, Bongrand P. Diffusion of microspheres in shear flow near a wall: use to measure binding rates between attached molecules. Biophys J 2001; 81: 25-42.

11. Krasik EF, Hammer DA. Asemianalytic model of leukocyte rolling. Biophys J 2004; 87: 2919-30.

12. Wierenga AM, Lenstra TAJ, Philipse AP. Aqueous dispersions of colloidal gibbsite platelets; synthesis, characterization and intrinsic viscosity measurements. Colloids Surf A-Physicochem Eng Aspects 1998; 134(3); 359-71.

13. Illing A, Unruh T, Koch MH. Investigation on particle self-assembly in solid lipid-based colloidal drug carriersystems. Pharm Res 2004; 21:592-7.

14. van Dillen T, van Blaaderen A, Polman A. Ion beam shaping of collagen assemblies. Mater Today 2004: 40-6.

15. Kohli P, Martin CR. Smart nanotubes for biotechnology. Curr Pharm Biotechnol 2005; 6(1): 35-47.16. Subramaniam AB, Abkarian M, Mahadevan L, Stone HA. Non-spherical bubbles. Nature 2005; 438: 930 .

17. Rolland JP, Maynor BW, Euliss LE, Exner AE, Denison GM, DeSimone J. Direct fabrication and harvesting of monodisperse, shape specific nano-biomaterials. J Am Chem Soc2005; 127:10096-100.18. Decuzzi P, Lee S, Bhushan B, Ferrari M. A theoretical model for the margin of particles within blood vessels. Ann Biomed Eng 2005; 33(2); 179-90.

19. Pozrikidis C. The motion of particles in the Hele-Shawcell. J Fluid Mech 1994; 261: 199-222.

20. Goldman AJ, Cox RG, Brenner H. Slow viscous motion of a sphere parallel to aplane wall. II. Couette flow. Chem EngSci 1967; 22: 653.

21. McQuarrie DA. Kinetics of small systems. J Chem Eng Phys1963; 38: 433-5.

22. Piper JW, Swerlick RA, Zhu C. Determining force dependence of two-dimensional receptor-ligand binding affinity by centrifugation. Biophys J 1998;74:492-513.

23. Shinde Patil VR, Campbell CJ, Yun YH, Slack SM, GoetzDJ. Particle diameter influences adhesion under flow. BiophysJ 2001; 80: 1733-43.

24. Gavze E, Shapiro M. Motion of inertial spheroidal particles in a shear flow near a solid wall with special application to aerosol transport in microgravity. J Fluid Mech1998; 371: 59-79.

25. Jain R.K. 2001. Delivery of molecular and cellular medicine to solid tumors. Advanced Drug Delivery Reviews. 46: 149-168.

26. Mollica F., R.K. Rakesh, and P.A. Netti. 2003. A model for temporal heterogeneities of tumor blood flow. Microvascular Research. 65: 56-60.

27. Hashizume H., P. Baluk, S. Morikawa, J.W. McLean, G. Thurston, S. Roberge, R.K. Jain, and D.M. McDonald. 2000. Openings between defective endothelial cells explain tumor vessel leakage. American Journal of 16 Pathology 4): 1363-1380.

28. Decuzzi P., F. Causa, and P.A. Netti. 2005. The effective dispersion of nanovectors within the microvasculature. Submitted on the Annals of Biomedical Engineering.

29. Netti, P.A., D.A.Berk, M.A.Swartz, A.J.Grodzinsky, and R.K.Jain.2000.Role of extracellular matrix assembly ininterstitial transport in solid tumors, Cancer Research.60:2497-2503.

30. Decuzzi, P.S. Lee, M. Decuzzi, and M. Ferrari. 2004. Adhesion of microfabricated particles on vascular endothelium: a parametric analysis, Annals of Biomedical Engineering. 32(6): 793-802.

31. Krasnici, S., A. Werner, M. E. Eichhorn, M. Schmitt-Sody, S. A. Pahernik, B. Sauer, B. Schulze, M. Teifel, U. Michaelis, K. Naujoks, and M. Dellian. 2003. Effect of the surface charge of liposomes on their uptake by angiogenic tumor vessels. Int. J. Cancer. 105(4): 561-567.

32. Gbadamosi, J.K., A.C.Hunter, and S.M.Moghimi. 2002. PEGylation of microspheres generates a heterogeneous population of particles with differential surface characteristic and biological performance, FEBS Lett.532(3): 338-344.

33. Rijnaarts, H.H.M., Norde, J.Lyklema, and A.Zehnder. 1999. DLVO and steric contributions to bacterial deposition inmedia of different ionic strengths. Colloids and Surfaces B: Biointerfaces, 14(1-4): 179-195.

34. Yu, Z.W., T.L. Calvert, and D. Leckbank. 1998. Molecular forces between membranes displaying neutral glycosphingolipids: Evidence for carbohydrate attraction. Biochemistry. 37: 1540-1550. , and R. Depieds. 1981. Nonspecific binding by macrophages: evaluation of the influence of medium-range electrostatic repulsion and short-range hydrophobic interaction. Immunol Commun 10: 35-43.

36. Israelachvili, J.1992. Intermolecular and surfaceforces, 2nd ed. Academic Press, New York.

37. Hsu, R., and P.Ganatos.1989.The motion of a rigid body in viscous fluid bounded by a plane wall.J.Fluid Mech.207:29-72.

38. Mege, J.L., C.Capo, A.M.Benoliel, and P.Bongrand.1987.Use of cell contour analysis to evaluate the affinity between macrophages and glutaraldehyde-treated erythrocytes.Biophys J.52(2):177-86.

39. Ganong, W.F. Review of medical physiology, 21st ed. Lange Medical Books/McGraw-Hill Medical Publishing Division, New York.

Claims (41)

1. the method for treatment or monitoring physiological situation comprises the experimenter that these needs are arranged is used the compositions that comprises the oblate spheroid particle that described oblate spheroid particle comprises at least a active agent of effective dose.

2. the process of claim 1 wherein that described at least a active agent comprises therapeutic agent.

3. the process of claim 1 wherein that described at least a active agent comprises preparation.

4. the process of claim 1 wherein that described particle comprises nano-porous materials.

5. the method for claim 4, wherein said particle comprises the nanoporous oxide material.

6. the method for claim 5, wherein said particle comprises nano-stephanoporate silicon dioxide.

7. the method for claim 4, wherein said nano-porous materials is a nano-structure porous silicon.

8. the process of claim 1 wherein that described particle comprises Biodegradable material.

9. the process of claim 1 wherein that described particle comprises at least a identification division that is positioned on the described particle surface.

10. the process of claim 1 wherein that described experimenter is a mammal.

11. the process of claim 1 wherein that described experimenter is the people.

12. comprise the compositions of bolt for fastening a door from outside spheroidal particle, described oblate spheroid particle comprises at least a active agent.

13. the compositions of claim 12, wherein said at least a active agent comprises therapeutic agent.

14. the compositions of claim 12, wherein said at least a active agent comprises preparation.

15. the compositions of claim 12, wherein said particle comprises nano-porous materials.

16. the compositions of claim 15, wherein said particle comprises the nanoporous oxide material.

17. the compositions of claim 16, wherein said particle comprises nano-stephanoporate silicon dioxide.

18. the compositions of claim 15, wherein said nano-porous materials is a nano-structure porous silicon.

19. the compositions of claim 12, wherein said particle comprises Biodegradable material.

20. the compositions of claim 12, wherein said particle comprise at least a identification division that is positioned on the described particle surface.

21. method, it comprises

(A) select to have surperficial target site, described surface has one or more firsts;

(B) select and the complementary second portion of first;

(C) select to pass through the shape that one or more form parameters define;

(D) determine the volume that adhere to of maximization, based on (i) selected one or more form parameters to target site; (ii) interactional one or more parameters between first and the second portion; The (iii) area density of first on target site; And

(E) make particle, the volume that it has selected basically shape and determines basically; And

(F) second portion is placed on the surface of particle.

22. the method for claim 21, wherein said target site are in endovascular site.

The site takes place 23. the method for claim 22, wherein said target site are blood vessels.

24. the method for claim 21, wherein said first is included in the receptor of expressing on the surface of described target site.

25. the method for claim 24, wherein said second portion comprise and the complementary part of described receptor, fit or antibody.

26. the method for claim 21, wherein said making comprise by the making by technology under pushing up extremely.

27. the method for claim 21, wherein said particle is a spheroidal particle, and one or more form parameters are spherical length-width ratios.

28. the particle of making according to the method for claim 21.

29. comprise in a large number the compositions of the particle of making according to the method for claim 21.

30. the compositions of claim 29, it comprises the suspension that contains described big quantity of material.

31. method, it comprises

(A) select to have surperficial target site, described surface has one or more firsts;

(B) select volume;

(C) select and the complementary second portion of described first;

(D) determine the shape of adhering to of maximization, based on (i) selected volume to target site; (ii) interactional parameter between first and the second portion; The (iii) area density of first on target site;

(E) make particle, it has shape and the selected basically volume of determining basically; And

(F) second portion is placed on the surface of particle.

32. the method for claim 31, wherein (B) comprises that the target of selecting active agent loads and loads based on described target and determine volume, and wherein said method also comprises active agent is loaded into particle.

33. the method for claim 31, wherein said active agent are therapeutic agent or preparation.

34. the method for claim 31, wherein said target site are in endovascular site.

The site takes place 35. the method for claim 34, wherein said target site are blood vessels.

36. the method for claim 34, wherein said first is included in the receptor of expressing on the surface of described target site.

37. the method for claim 36, wherein said second portion comprise and the complementary second portion of described receptor.

38. the method for claim 31, wherein said making comprise by the making by technology under pushing up extremely.

39. the method for claim 31, wherein said particle is a spheroidal particle, and wherein saidly determines to adhere to maximum shape and comprise and determine to adhere to the maximum spherical length-width ratio.

40. the particle of making according to the method for claim 31.

41. comprise in a large number the compositions of the particle of making according to the method for claim 31.

Images