Mapping nanoparticles

Abstract

To provide a nanoparticulate material suitable for administration to a subject, which accurately identifies the margins of solid tumours.SOLUTION: Provided is a nanoparticulate material having bound to its surface: (a) copolymeric steric stabiliser that promotes dispersion of the nanoparticulate material in a liquid, wherein the copolymeric steric stabiliser comprises (i) an anchoring polymer segment having one or more binding groups that bind the copolymeric steric stabiliser to the nanoparticulate material, and (ii) a steric stabilising polymer segment that is different from the anchoring polymer segment, and (b) copolymeric mapping moiety comprising (i) an anchoring polymer segment having one or more binding groups that bind the copolymeric mapping moiety to the nanoparticulate material, (ii) one or more mapping groups comprising an agent that specifically binds to fibroblast activation protein (FAP), and (iii) a coupling polymer segment that is different from the anchoring polymer segment.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This application generally relates to nanoparticles, compositions containing same, and methods for mapping, diagnosis, and treatment. and their use in therapeutic applications. [Background technology]

[0002] Surgical tumor resection is the standard treatment for many cancers, especially when the disease is localized to a single solid tumor. However, surgery is invasive and does not identify residual neoplastic tissue. may result in positive surgical margins, which correlates with local recurrence and poor patient outcomes. Furthermore, accurate identification of tumor margins is crucial for the treatment of cancers with external beam radiotherapy, brachytherapy, and and focal therapy, each of which is based on the location and extent (e.g., volume) of the tumor. The best possible characterization is important for the effectiveness of required non-surgical treatment modalities .

[0003] For example, radical prostatectomy is the standard treatment for aggressive and intermediate-stage prostate cancer. However, after this surgical intervention, approximately 20% of patients experience urinary incontinence, and Approximately 70% of men experience erectile dysfunction. These serious side effects limit life expectancy and progress. Men with slow-growing, low-risk disease should be given "watchful waiting" before undergoing surgery. Alternatively, for some patients, definitive treatment is recommended. External beam radiation therapy, brachytherapy, and focal therapy before prostatectomy Such treatments are associated with fewer side effects, but are not as effective as surgical treatments. It is not as effective as partial or complete tumor resection. The effectiveness of these alternative treatment modalities The reduction factor may provide sufficient spatial resolution to identify the borders and margins of the primary tumor. This is a limitation of existing medical imaging. For example, prostate-specific membrane antigen (PSMA) positron emission tomography / Computed tomography (PET-PSMA) may underestimate tumor volume by 9-15% Multiparametric magnetic resonance imaging (mpMRI) has been shown to reduce tumor volume by 11-20%. Due to imaging limitations, focal therapy guidelines In this case, the margins of the resection area surrounding the identified lesion should be extended up to 10 mm. However, even if the resection area is expanded, a 20-40% recurrence rate at the margin has been reported. Organ preservation and limiting normal tissue toxicity are important, e.g., glioblastoma and pancreatic cancer. Similar issues arise with other cancers, including but not limited to cancer. Summary of the Invention [Problem to be solved by the invention]

[0004] Therefore, by providing improved pre- and / or intra-operative guidance To improve surgical resection or to use less invasive treatment modalities, e.g., cryoablation ablation, focal laser ablation, and high-frequency ultrasound ablation. Focal therapy, photodynamic therapy, high-dose rate and low-dose rate brachytherapy, protocoagulation therapy, particle radiation therapy, such as ion and carbon ion radiotherapy, and intensity-modulated radiation therapy (IMRT) , image-guided radiation therapy (IGRT), hypofractionated radiation therapy, and ultra-high dose rate radiation therapy. External beam radiotherapy can more precisely target the entire tumor volume, reducing the risk of non-targeted tissue New materials and methods to accurately identify the margins of solid tumors, potentially reducing tissue exposure There remains a need to develop novel anti-inflammatory and anti-inflammatory compositions. [Means for solving the problem]

[0005] Surprisingly, the nanoparticle material according to the present invention selectively accumulates within the tumor microenvironment and It has been found that this method can map the margins of solid tumors. Although it is not desired to do so, the function of the copolymer steric stabilizer and the copolymer mapping moiety is In combination, the nanoparticle materials described herein can be used to target cells within the tumor microenvironment, e.g. For example, selective for fibroblast activation protein (FAP) expressed by tumor-associated stromal cells to be at least partially bonded to the

[0006] Thus, in one aspect described herein, a nanoparticle material suitable for administration to a subject is provided. and (a) a copolymer steric stabilizer that facilitates dispersion of the nanoparticle material in a liquid. (i) one or more stabilizers that bind the copolymer steric stabilizer to the nanoparticle material; and (ii) an anchoring polymer segment having a linking group of the anchoring polymer segment. (b) a copolymer steric stabilizer comprising a steric stabilizing polymer segment different from the copolymer steric stabilizer; (i) coupling said copolymer mapping moiety to said nanoparticle material; (ii) an immobilized polymer segment having one or more binding groups for binding to fibroblasts; one or more mapping groups comprising an agent that specifically binds to a fibrinogen activator protein (FAP) and and (iii) a coupling polymer segment different from the immobilizing polymer segment. and coupling said immobilization polymer segment to said one or more mapping groups. and a copolymer mapping portion comprising a coupling polymer segment. A nanoparticle material is provided.

[0007] The nanoparticle materials according to the present invention advantageously exhibit improved blood half-life circulation and are distributed throughout the body. Supports injection and / or does not substantially degrade in vivo.

[0008] Without wishing to be bound by theory, nanoparticle materials are It is believed that such advantageous properties are exhibited through the copolymer component attached to the surface. These copolymer components contain copolymer steric stabilizers in combination with copolymer mapping moieties. Both the copolymer steric stabilizer and the copolymer mapping moiety are The polymeric material comprises an immobilizing polymer segment that binds the nanoparticle material to the immobilizing polymer segment. The nanoparticles advantageously remain immobilized on the nanoparticle material when in, for example, an in vivo fluid environment. Highly effective in maintaining both the copolymer mapping moiety and the copolymer steric stabilizer This shows that nanoparticle materials can be dispersed in the liquid environment of a living body. Those skilled in the art will appreciate that the aggregation of nanoparticle materials in an in vivo liquid environment is facilitated. It will be appreciated that aggregation can be detrimental in diagnostic and therapeutic applications.

[0009] Again, without wishing to be bound by theory, the copolymer mapping portion It acts synergistically with the effect of improving dispersion in the in vivo liquid environment, etc., and It is believed that this will allow for ameliorating the accumulation of particulate material in the environment. In response, the nanoparticle material is coupled to the copolymer mapping moiety. They essentially accumulate in the tumor microenvironment similarly to other proteins, but are advantageously selected to perform a given task. For example, nanoparticle materials can be used in magnetic particle imaging (MPI) and nuclear magnetic resonance imaging (NMR). Imaging methods (MRI), e.g., low-field MRI, MRI-guided external beam radiation therapy, MRI-guided MRI-guided focal ablation, MRI / ultrasound fusion focal ablation, MRI-guided guided biopsy, MRI / ultrasound fusion guided biopsy, MRI guided surgery, MRI guided brachytherapy, M For applications such as RI-guided infrared camera-guided biopsy or treatment and photoacoustic-guided biopsy or treatment It can be provided in the form of a magnetic nanoparticle material for use.

[0010] Thus, the nanoparticle material according to the present invention can be used to map tumor margins in a subject and to identify tumor lesions. This improves conventional methods for determining the location and extent (e.g., volume) of tumors and therapeutic applications. It was found to be particularly effective in this regard.

[0011] Those skilled in the art will appreciate that the application of targeted nanoparticle materials in vivo may be advantageously utilized to prevent unwanted protein adsorption. This can lead to the formation of a so-called protein corona, which can cause binding at the target site. It will be appreciated that such protein adsorption may result in a decrease in binding efficiency. This may not only reduce the risk of infection but also result in the accumulation of nanoparticle materials at the target site. Surprisingly, copolymer steric stabilizers and copolymer mapping The combined use of PEGs can also advantageously reduce the effects of detrimental protein adsorption. , which was found to improve the accumulation efficiency of nanoparticle materials within the tumor microenvironment.

[0012] In one embodiment, the nanoparticle material has on its surface (c) a copolymeric luminescent moiety, (i) an immobilized polymer having one or more linking groups that attach the polymeric light-emitting moiety to the nanoparticle material; (ii) a photoreceptor segment that allows visualization of the location of the nanoparticle material in vivo. (iii) one or more luminescent groups for emitting an acoustic signal in response to light or light; The coupling polymer segment is different from the immobilized polymer segment. a coupling polymer segment that couples the cation to one or more luminescent groups. The copolymer contains a light-emitting moiety attached thereto.

[0013] In one embodiment, the steric stabilizing polymer segment is a polyacrylamide-copolymer. Polyacrylamide-co-polyalkylene oxide block copolymer block copolymer).

[0014] In another embodiment, the coupling polymer segment is made from polyacrylamide. It is composed.

[0015] In a further embodiment, the steric stabilizing polymer segment is a polyacrylamide- Co-polyalkylene oxide block copolymers, including coupling polymer segments The material is composed of polyacrylamide.

[0016] In one embodiment, the steric stabilizing polymer segment comprises 10 to 70 polymerized monomers. -residue units.

[0017] In another embodiment, the coupling polymer segment comprises 15 to 100 polymerized units. It has monomer residue units.

[0018] In a further embodiment, the steric stabilizing polymer segment comprises 10 to 70 polymerized units. The coupling polymer segment has 15 to 100 polymerized units. It has monomer residue units.

[0019] In one embodiment, the nanoparticle material is a magnetic nanoparticle material.

[0020] As described elsewhere herein, nanoparticle materials are bioavailable to cells within the tumor microenvironment. It selectively binds to fibroblast activation protein (FAP) expressed by the IL-1 receptor.

[0021] In one embodiment, the cells within the tumor microenvironment are tumor-associated stromal cells.

[0022] In further embodiments, the tumor-associated stromal cells are selected from the group consisting of fibroblasts, pericytes, adipocytes, mesenchymal In another embodiment, the endothelial cells are selected from mesenchymal stromal cells (MSCs), endothelial cells, and combinations thereof. The tumor-associated stromal cells are selected from pericytes, endothelial cells, and combinations thereof.

[0023] In one embodiment, agents that specifically bind to FAP include small molecule inhibitors and antibodies or is selected from antigen-binding fragments thereof. In another embodiment, the small molecule inhibitor is It is a FAP inhibitor.

[0024] In further embodiments, the one or more luminescent groups are chemiluminescent groups, electroluminescent groups, photoluminescent groups, The luminescent group is selected from a luminescent group, a radioluminescent group and a thermoluminescent group.

[0025] In another aspect disclosed herein, there is provided a composition suitable for administration to a subject, comprising: A composition including the nanoparticle material according to the invention is provided.

[0026] In one embodiment, the composition comprises a pharmaceutically acceptable liquid carrier.

[0027] In another embodiment disclosed herein, when performing a therapeutic or diagnostic application on a subject The use of the nanoparticle material or composition according to the invention is provided.

[0028] Examples of suitable therapeutic or diagnostic applications include magnetic particle imaging (MPI), nuclear magnetic resonance imaging (NMR), and magnetic resonance imaging (MRI). Magnetic resonance imaging (MRI), MRI-guided external beam radiation therapy, MRI-guided focal ablation MRI / ultrasound fusion focal ablation, MRI guided biopsy, MRI / Ultrasound fusion-guided biopsy, MRI-guided surgery, MRI-guided brachytherapy, MRI-guided infrared camera Laser-guided biopsy or therapy and photoacoustic-guided biopsy or therapy are mentioned.

[0029] The nanoparticle material or composition according to the present invention may be used in ultrasound, MRI / ultrasound, X-ray, optical imaging, ging, computed tomography (CT), single photon emission computed tomography (SP) ECT), Positron Emission Tomography (PET), Fluorescence Resonance Energy Transfer (FRET) and In combination with in vivo imaging techniques, including but not limited to magnetic resonance imaging (MRI) It can be used in combination.

[0030] In another aspect disclosed herein, a nanoparticle material according to the present invention for in vivo imaging is provided. Alternatively, uses of the composition are provided.

[0031] In another aspect disclosed herein, a nanoparticle according to the present invention is provided for use in in vivo imaging. A particulate material or composition is provided.

[0032] In another aspect disclosed herein, a nanoparticle material according to the present invention for detecting cancer is provided. Use of the material or composition is provided.

[0033] In another aspect disclosed herein, a nanoparticle according to the present invention is provided for use in detecting cancer. A particulate material or composition is provided.

[0034] In one embodiment, the cancer is prostate cancer, glioblastoma multiforme, glioma, or pancreatic cancer. , colorectal cancer, breast cancer, head and neck cancer, gastric cancer, esophageal cancer, ovarian cancer, sarcoma and lung cancer In another embodiment, the cancer is prostate cancer.

[0035] In another aspect disclosed herein, a method for mapping the tumor microenvironment is provided. The use of the nanoparticle material or composition is provided.

[0036] In another aspect disclosed herein, the present invention provides a method for mapping the tumor microenvironment. According to the invention, nanoparticle materials or compositions are provided.

[0037] In one aspect disclosed herein, a. administering to a subject a nanoparticle material or composition according to the present invention; b. detecting the nanoparticle material; a method for mapping tumor margins in a subject, the method comprising: A method is provided for accumulating tumor cells in a single cell, thereby mapping tumor margins.

[0038] In one embodiment, tumor margins are mapped in situ. can be mapped in situ before or after.

[0039] In one embodiment, the method includes measuring the clinical target volume (CTV) and / or the muscle prior to administering the treatment. Further comprising determining the ocular target volume (GTV).

[0040] In one aspect disclosed herein, a method for treating cancer in a subject in need thereof is provided. 1. A method for: a. administering to a subject a nanoparticle material or composition according to the present invention; b. detecting sites of accumulation of nanoparticle material; c. administering to the site of detection of the nanoparticle material in step (b) an effective amount for treating said cancer. and A method is provided, comprising:

[0041] In one embodiment, the treatment is surgery, radiation therapy, brachytherapy, photodynamic therapy, Photothermal therapy, (cryoablation, focal laser ablation and high frequency ultrasound Focal ablation therapy, chemotherapy, immunotherapy and The combination is selected from the following:

[0042] In one embodiment, the nanoparticle material is capable of detecting magnetic resonance imaging (MRI), ultrasound, x-ray, optical Imaging, Fluorescence Imaging, Computed Tomography (CT), Single Photon Emission Computing PET (Positron Emission Tomography) and Fluorescence Resonance Energy The signal is detected using an imaging technique selected from: fluorescence emission transfer (FRET);

[0043] In one embodiment, the method further comprises administering a clinical target volume prior to administering the treatment according to step (c). This includes determining the carotid target volume (CTV) and / or gross target volume (GTV).

[0044] In one aspect disclosed herein, a. administering to a subject a nanoparticle material or composition according to the present invention; b. detecting the nanoparticle material; 1. A method for diagnosing cancer, comprising: Detection of nanoparticle material accumulated in tissue (such as vascular tissue) of a subject can indicate whether the subject has cancer. A method is provided that shows that

[0045] Further aspects and embodiments of the present invention are outlined and discussed in more detail below. do.

[0046] Embodiments of the present disclosure will now be described with reference to the accompanying non-limiting figures. [Brief explanation of the drawings]

[0047] [Figure 1] FIG. 1 is a schematic diagram illustrating a nanoparticle material according to the present invention. [Figure 2] FIG. 2 shows the chemical structure of a "long" copolymer steric stabilizer according to the present invention. [Figure 3] FIG. 3 shows the results of dynamic light scattering (DLS) measurements of dispersed nanoparticle materials having prostate-specific membrane antigen (PSMA) targeting groups attached to their surfaces. [Figure 4] FIG. 4 shows the chemical structure of a "long" copolymer mapping portion according to the present invention. [Figure 5] FIG. 5 shows the results of dynamic light scattering (DLS) measurements of dispersed nanoparticle materials having fibroblast activation protein inhibitor (FAPI) targeting groups attached to their surfaces. [Figure 6] FIG. 6 shows the 3T-MRI response of an orthotopic prostate tumor resection specimen, demonstrating that the nanoparticle material of the present invention has a strong MRI response compared to the weak MRI response from the comparative nanoparticle material shown in FIG. [Figure 7] FIG. 7 shows a pathology slide from an orthotopic tumor resection specimen, demonstrating that the nanoparticle material of the present invention has strong Prussian blue iron staining on the tumor surface, whereas the comparative nanoparticle material shown in FIG. 5 has far fewer nanoparticles on the tumor surface with iron blue staining. [Figure 8]FIG. 8 shows a 14.7T-MRI of a whole mouse body with negative contrast of nanoparticle material according to the present invention on the surface of an orthotopic prostate model. [Figure 9] FIG. 9 shows the chemical structure of a "short" copolymer mapping portion according to the present invention. [Figure 10] FIG. 10 shows the chemical structures of "short" copolymer steric stabilizers according to the present invention. [Figure 11] FIG. 11 shows a comparison of cellular uptake of small FAP-mapping magnetic nanoparticles (maghemite) and non-targeted small magnetic nanoparticles in FAP-expressing cell lines. [Figure 12] FIG. 12 shows a comparison of cellular uptake of small copolymer FAP-mapping magnetic nanoparticles (maghemite) and single polymer FAP-mapping magnetic nanoparticles in FAP-expressing cell lines in the presence and absence of fetal bovine serum (FBS). [Figure 13] FIG. 13 shows a comparison of cellular uptake of small FAP-mapping magnetic nanoparticles (magnetite) and non-targeted small magnetic nanoparticles (magnetite) in FAP-expressing cell lines. DETAILED DESCRIPTION OF THE INVENTION

[0048] In this specification, some terms are used that are well known to those skilled in the art. To do this, we define some terms.

[0049] The term "subject" means either an animal or a human subject. Long-legged animals, livestock animals (such as cattle, horses, sheep, pigs and goats), companion animals ( dogs, cats, rabbits and guinea pigs) and captive wild animals (raised in zoo environments) (including those that are used in the manufacture of rabbits, mice, rats, guinea pigs and hamsters) Laboratory animals such as rats are also contemplated as they may provide a convenient test system. In the present invention, the subject is a human subject.

[0050] A nanoparticle material or composition according to the invention is "suitable" for administration to a subject if: Administration of the composition to a subject results in unacceptable toxicity, such as allergic reactions and disease states. This means that it does not result in

[0051] "Administering" a nanoparticle material or composition to a subject means that the nanoparticle material or composition This means that the particulate material is provided in a manner that allows it to be delivered to a subject. This is generally, but not limited to, oral, parenteral (subcutaneous, intradermal, intramuscular, intravenous, intracerebral, intranasal) , intrathecal and intraspinal), inhalation (nebulization, etc.), topical, rectal and vaginal modes The nanoparticle material or composition may also be used to target one or more cells within and / or at the tumor. The drug may be administered directly into the tissue adjacent to the segment, or directly into the blood vessel. stomach.

[0052] "Pharmacologically acceptable" means suitable for administration to a subject. However, administration of related substances to subjects can result in unacceptable toxicity, such as allergic reactions and disease states. Do not bring it about.

[0053] While merely a guide, those skilled in the art will understand that "pharmacologically acceptable" is a term that is acceptable to a wide variety of people, including those in the art who are federally or state An entity approved by a government regulatory agency, or the United States Pharmacopoeia or Animals, more specifically are considered to be entities listed in other generally accepted pharmacopeias for human use. That's fine.

[0054] However, one of skill in the art would understand that administering nanoparticle materials or compositions according to the present invention to a subject and whether it or its constituents are considered pharmacologically acceptable. It will be appreciated that the extent to which the composition is effective will depend in part on the mode of administration selected. whether the composition or its components are suitable for administration to a subject or are pharmacologically acceptable. When assessing whether a drug is effective, the mode of administration may need to be considered.

[0055] The nanoparticle material is "dispersed throughout" the liquid carrier if the nanoparticle material is dispersed in a manner that is consistent with the particle material. The dispersed phase is dispersed throughout the liquid carrier, which itself exists as a continuous liquid medium or phase. That is, the composition according to the present invention is a liquid carrier of nanoparticle material. It may be described as involving suspension or dispersion throughout the body.

[0056] As used herein, the term "liquid" in the context of a liquid carrier refers to a medium. Thus, the nanoparticle material is dispersed throughout the composition, and the intended use of the composition of the present invention is The term "medium" is intended to mean a medium that is at least in a liquid state at the temperature of use. The liquid carrier is such that, in the absence of a stabilizer, the particulate material dispersed throughout the carrier is flocculated from the carrier. A substance is considered to be in a "liquid" state if it is capable of settling or settling to form a precipitate. That is, a "liquid" is a medium in which particulate material can move relatively freely. It is thought that...

[0057] The liquid carrier used in accordance with the present invention may be composed of one or more different liquids. Suitable pharmaceutical acceptable liquid carriers are described in Martin, ed., Remington's Ph.D. "Archemeutical Sciences", 18th Edition, Mack Publish ing Co., Easton, Pennsylvania (1990) and water, Oils of petroleum, animal, vegetable, mineral, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil Other liquid carriers include, but are not limited to, sterilizable liquids such as oils containing Examples include methylene glycol, propylene glycol, polyethylene glycol, and poly Examples include propylene glycol, ethanol, isopropyl alcohol, and benzyl alcohol. Water or soluble saline and aqueous dextrose and glycerol solutions are shown. The liquid is preferably used as a liquid carrier, particularly as a liquid carrier for injectable solutions.

[0058] The compositions according to the present invention may contain one or more pharmacologically acceptable additives known to those skilled in the art. For example, the liquid carrier may contain wetting agents, antifoaming agents, surfactants, buffers, electrolytes, preservatives, One or more additives such as coloring agents, flavoring agents and sweetening agents may be included.

[0059] The specific properties of the liquid carrier and any additives, if any, will depend on the intended use of the composition. Those skilled in the art will appreciate that the liquid carrier and formulation suitable for the intended use of the composition will be readily apparent to those skilled in the art. The supplier will be able to select the composition and additives (if any).

[0060] The nanoparticle material may be administered in a therapeutically or diagnostically effective amount, where appropriate. An effective amount is an amount that, when administered according to a desired dosing regimen, produces a desired therapeutic or diagnostic effect, e.g., For example, by the onset or progression of the particular condition being treated and / or evaluated. , alleviating the symptoms thereof, preventing or delaying the onset thereof, inhibiting or delaying the progression thereof, diagnosing thereof It is intended to include an amount that achieves one or more of the following: abrogation, or its cessation or reversal. .

[0061] The appropriate dosage and schedule to achieve this will be determined by your physician. the specific condition being treated or diagnosed, the severity of the condition, and the general age of the subject It may depend on age, health and weight.

[0062] Administration may be at intervals of minutes, hours, days, weeks, months, or years, or any of these. The dosage of the particulate material itself may be 1 Within the range of approximately 0.1 ng / kg to 1 g / kg of body weight per dose The dosage may be within the range of 1 μg to 1 g per kg of body weight per administration. For example, the dose per administration may be in the range of 1 mg to 1 g per kg of body weight. In this form, the dosage ranges from 1 mg to 500 mg per kg of body weight per administration. In another embodiment, the dosage may be within 1 kg of body weight per administration. In yet another embodiment, the dosage may range from 100 mg to 250 mg per administration. Within the range of 1 mg to 100 mg per kg of body weight, for example, The maximum amount of steroids administered per kg of body weight may be up to 50 mg.

[0063] The nanoparticle material or composition according to the invention may be administered in a single dose or in multiple doses.

[0064] When nanoparticulate materials or compositions according to the invention are suitable for parenteral administration, they are generally , antioxidants, buffers, bactericides or solutes that make the composition isotonic with the blood of the intended subject. In the form of an aqueous or non-aqueous isotonic sterile injection solution which may contain one or more of: Such compositions may be presented in unit-dose or multi-dose sealed containers, such as ampoules and the like. It may be provided in a vial.

[0065] Upon administration, the nanoparticle material or composition according to the invention may be diluted in vivo. For example, dilution may occur when administered orally or parenterally. The liquid carrier can be diluted very well in vivo, so that the surrounding liquid in which the particulate material is dispersed is The body environment is more like an in vivo fluid (i.e., a biological fluid / fluid within the subject) than the original fluid carrier. For example, when administered parenterally, the nanoparticle material will reflect the original composition. It would be more accurate to describe it as being dispersed throughout the blood rather than in a liquid carrier. Under certain circumstances, the nanoparticle material may be transported in an in vivo fluid carrier (i.e., a biological fluid / fluid within the subject). It may be convenient to refer to the composition according to the invention as being dispersed throughout the Except for the differences in composition between the liquid carrier and the in vivo liquid carrier, the liquid carrier of the composition is The statements also apply generally to in vivo liquid carriers.

[0066] As used herein, the term "tumor microenvironment" refers to the tissue surrounding a tumor. and / or a heterogeneous population of non-cancerous cells infiltrating the tumor, which It is of great importance to the function, physiology, and metastasis of tumors. Contains a variety of different cell types that can vary based on size, location, type and stage Illustrative examples include fibroblasts, pericytes, adipocytes, and mesenchymal stromal cells ( It will be understood that the tumor microenvironment includes cells such as MSCs, cancer cells, and endothelial cells. These cells are non-cancerous, but tumors recruit and / or control such cells to cause cancer. Therefore, the tumor microenvironment provides a favorable environment for promoting the growth of Cells that are involved are sometimes referred to as "cancer-associated" or "tumor-associated."

[0067] In one embodiment, the nanoparticle material disclosed herein is metabolized by cells within the tumor microenvironment. It selectively binds to fibroblast activation protein (FAP) expressed on the surface of the fibroblasts.

[0068] In one embodiment, the cells within the tumor microenvironment are tumor-associated stromal cells.

[0069] In one embodiment, the tumor-associated stromal cells are fibroblasts, pericytes, adipocytes, mesenchymal stroma cells. The cells are selected from the group consisting of MSCs, cancer cells, endothelial cells, and combinations thereof. In some embodiments, the tumor-associated stromal cells are selected from pericytes, endothelial cells, and combinations thereof. .

[0070] The terms "about" or "approximately" refer to an acceptable range for a particular value as determined by one of ordinary skill in the art. within tolerance, which refers to how the value is measured or determined, e.g., by the measurement system For example, "about" means up to 20%, preferably up to 10%, of a given value. % or less, more preferably 5% or less, and even more preferably 1% or less. Alternatively, particularly with respect to biological systems or processes, the term can be used to describe the It can mean within 10 times, preferably within 5 times, more preferably within 2 times. Unless otherwise specified, the term "about" refers to a range of acceptable error for a particular value, e.g., ±1-2 It means 0%, preferably ±1 to 10%, and more preferably ±1 to 5%.

[0071] When a range of values is provided, each intervening value between the upper and lower limits of the range and its description It is understood that any other stated or intervening value within the stated ranges is encompassed within the disclosure. The upper and lower limits of these narrower ranges may independently be included within the narrower ranges. Any specifically excluded within the stated ranges is also encompassed within this disclosure. When the stated range includes one or both of the limits, Ranges excluding either or both of these inclusive limits are also included within the disclosure.

[0072] A phrase referring to "at least one" of a list of items refers to any combination of those items. , contains a single member. As an example, "at least one of a, b, or c" means intended to encompass a, b, c, ab, ac, bc, and abc .

[0073] To aid in the description of nanoparticle materials according to the present invention, reference is made to Figure 1. The nanoparticle material itself (20) is made of iron oxide nanoparticles. The surface of the nanoparticle material (20) is represented by (i) features (30), Copolymer mapping portions collectively represented by (40) and (50) and ( ii) copolymer steric stabilizers, collectively represented by forms (30) and (60), are formed The copolymer mapping portion is composed of an immobilized polymer segment (30) and a colorimetric a coupling polymer segment (40) and a coupling polymer segment (41) The coupling polymer segment (40) is different from the coupling polymer segment (40). The copolymer is sterically stabilized by coupling one or more mapping groups (50) to the copolymer. The agent is an immobilizing polymer segment (3) attached to a steric stabilizing polymer segment (60). 0), and the anchoring polymer segment (30) comprises a steric stabilizing polymer segment (6 The coupling polymer segment (40) is a steric stabilizing polymer. More polymerized monomer residue units than segment (60) residue units), thereby providing a steric stabilizing polymer segment (60) with a Thus, one or more mapping groups (50) extend a greater distance from the surface of the nanoparticle material (20). It becomes possible to extend it.

[0074] A particulate material is a "nano" particulate material if at least one of its dimensions is less than 100 nm. or less than about 75 nm, or less than about 50 nm, or less than about 30 nm. In one embodiment, all dimensions of the nanoparticle material are less than 100 nm, or about 7 It is less than 5 nm, or less than about 50 nm, or less than about 30 nm.

[0075] The nanoparticle material may be in the form of primary particles or in the form of aggregates of primary particles. In terms of morphology, the nanoparticulate material is in the form of primary particles.

[0076] For the avoidance of doubt, references herein to the "size" of nanoparticle materials include: It indicates the average size of particles (at least about 50% by number) based on the largest dimension of a given particle. It is intended to do so.

[0077] The size of the nanoparticle material itself is determined herein by transmission electron microscopy (TEM). It is determined.

[0078] For the avoidance of doubt, when nanoparticle material is in the form of an aggregate of primary particles, References to the size of such materials refer to the largest dimension of the aggregate, not the primary particles forming the aggregate. It is intended to be a reference to

[0079] In certain embodiments, the nanoparticle material is approximately In certain embodiments, the nanoparticle material has a size of less than 50 nm. or about 5 nm to about 30 nm in all dimensions, or about 5 nm to about 20 nm, or The size is in the range of about 8 nm to about 15 nm.

[0080] In one embodiment, the nanoparticle material is about 6, 8, 10, 15, 20, 30, 40, 50, The size is 60, 70, 80, 90 or 100 nm.

[0081] Nanoparticle materials are generally solid at temperatures typically encountered in their intended applications. During use in its intended application and during storage prior to use, Considering the temperatures to which the nanoparticle material or composition may be exposed, at least The outer surface is generally in a solid state at least below about 40°C, preferably below about 50°C. Nanoparticle materials, of course, have such solid state compositions throughout. and in some embodiments, such solid state compositions can be used throughout. (i.e., a solid nanoparticle material).

[0082] Apart from having pharmaceutical or diagnostic utility, there are no particular limitations on the composition of the nanoparticle material. No. The nanoparticle material may have an organic or inorganic composition or a combination thereof. The nanoparticle material may be a pharmaceutically active compound (e.g., a drug), a metal, an alloy, a metal salt, a metal complexes, metal oxides, radioisotopes, luminescent compounds or groups and / or combinations thereof The composition may be selected from or include a combination of:

[0083] Suitable nanoparticle materials include gold, silver and their salts, complexes or oxides, calcium carbonate, Sulfur dioxide, barium sulfate, bismuth sulfide, iron, iron oxide, chromium oxide, cobalt oxide, manganese oxide Gun, iron oxyhydroxide, chromium oxyhydroxide, cobalt oxyhydroxide, manganese oxyhydroxide Cancer, chromium dioxide, other transition metal oxides (Auger electron emitters, alpha emitters, cations) radioisotopes selected from electron emitters and beta emitters, and combinations thereof; may include:

[0084] Examples of Auger electron emitters include: 51 Cr, 67 Ga, 71 Ge, 75 Se, 77 B r, 80m Br, 99m Tc, 103 Pd, 103m Rh, 111 In, 113m In, 115m In, 117m Sn, 119 Sb, 123 I, 125 I, 131 Cs, 161 H o, 165 Er, 193m Pt, 195m Pt, 201 Tl and 203 Pb is listed do.

[0085] Examples of alpha emitters include: 211 At and 213 Bi is listed.

[0086] Examples of beta emitters include: 191 Os, 35 S, 33 P, 45 Ca, 199 Au, 1 69 Er, 67 Cu, 47 Sc, 177 Lu, 161 Tb and 105 Low energy such as Rh Energy beta emitters; 131 I, 153 Sm, 77 As, 143 Pr, 198 Au, 159 G d. 109 Pd, 186 Re, 111 Ag and 149 Medium energy beta emitters such as Pm ; and 165 Dy, 89 Sr, 32 P, 166 Ho, 188 Re, 114m In, 1 42 Pr, 90 Y and 76 Examples include high-energy beta emitters such as As.

[0087] Examples of radioisotopes that can be used in radiation therapy include: 32 P, 153m S, 90 Y, 125 I, 192 Ir, 103 Pd, 111 In, 166 Ho and 213 Bi raised can be.

[0088] Examples of radioisotopes that can be used as diagnostic agents include: 99m Tc, 67 Ga, 64 Cu, 89 Zr and 18 F is listed.

[0089] Examples of positron emitters that can be used as diagnostic agents include gallium-68, copper-64, zinc-68, and zinc-68. Copper-89, yttrium-86, rubidium-82, scandium-44, and several isotopes of copper Examples include terbium-182, gallium-66, and cobalt-55.

[0090] When radioisotopes are used, the radionuclides may be used as the nanoparticle material itself. or may be combined with one or more other suitable nanoparticle materials. The nanoparticle material may include one or more radioisotopes, for example: 67 Ga iron oxide particles It may also be used in combination with other materials.

[0091] In some embodiments, the nanoparticle material exhibits magnetic properties. Such magnetic nanoparticle materials are It may exhibit magnetic, ferrimagnetic or superparamagnetic properties.

[0092] In one embodiment, the nanoparticle material exhibits superparamagnetism.

[0093] The nanoparticle material may be made from or include a magnetic material.

[0094] Examples of suitable magnetic materials include iron, nickel, chromium, cobalt, gadolinium, and manganese. Cancer, any oxide or oxyhydroxide of the foregoing, and any mixture of the foregoing In certain embodiments, the magnetic nanoparticles include, but are not limited to, iron and Suitable iron oxide magnetic materials include magnetic Hematite (γ-Fe2O3) and magnetite (Fe3O4) are mentioned.

[0095] In one embodiment, the magnetic nanoparticle material is selected from the group consisting of iron, nickel, chromium, cobalt, gadolinium, and the like. One or more of aluminum, manganese, and their oxides or oxyhydroxides include.

[0096] In another embodiment, the magnetic nanoparticle material is iron (Fe), maghemite (γ-Fe2O3), Magnetite (Fe3O4) or combinations thereof.

[0097] In some embodiments, the magnetic nanoparticle material has a particle size of less than about 30 nm, for example, about 1 nm to Magnetite (Fe3O4) or maghemite (γ-Fe2O3) of approximately 20 nm, or includes it.

[0098] Magnetic nanoparticle materials consist of a magnetic metal such as maghemite (γ-Fe2O3) shell around a core material. It may also be in the form of a metal such as iron surrounded by a metal oxide shell.

[0099] In some embodiments, the magnetic nanoparticle material has the general formula MO·Fe 2 O 3 , where M is F e, Co, Ni, Mn, Be, Mg, Ca, Ba, Sr, Cu, Zn, Pt, Gd, etc. Ferrites of divalent metals or their mixtures, or of the general formula MO 6FeO 3, where M is a large divalent ion, metallic iron, cobalt, or nickel. The magnetic nanoparticle material is or contains a mineral oxide. It may consist of Cr, Co or Gd, or their oxides or oxyhydroxides. Alternatively, the magnetic nanoparticle material may be a mixture of any of these.

[0100] For some applications, it may be desirable to use magnetic nanoparticle materials that exhibit superparamagnetic properties. As used herein, the term "superparamagnetic" refers to the following properties: i) coercivity, (ii) remanence, or (iii) when the rate of change of the applied magnetic field is quasi-static It is intended to mean a magnetic material that does not have a hysteresis loop.

[0101] In some embodiments, the nanoparticle material is or includes a luminescent material.

[0102] Such luminescent materials include chemiluminescent (e.g., bioluminescent, electrochemiluminescent, thermoluminescent) electroluminescent, photoluminescent (e.g., fluorescent or phosphorescent), They may be radioluminescent or thermoluminescent. Examples of such luminescent materials include: , including the luminescent groups described herein.

[0103] The luminescent material is a nano particular material. For example, the luminescent material may be combined with another material to form a nanoparticle material. It may be encapsulated within the material.

[0104] The nanoparticle material according to the present invention has a surface on which a copolymer that promotes dispersion of the nanoparticle material in a liquid is formed. The liquid may be a carrier liquid or a biocompatible material as described herein. "Promoted" in that context means that the copolymer is an intrinsic liquid in the absence of a steric stabilizer. The nanoparticle material may flocculate, aggregate, or precipitate from the liquid. This means that the copolymer steric stabilizer acts to stabilize the nanoparticle material in the liquid. It functions to keep the particles dispersed.

[0105] The copolymer is a "steric" stabilizer, meaning that the dispersion of nanoparticle materials in a liquid is stabilized by steric repulsion. That said, copolymer steric stabilizers are also effective in reducing the amount of nanoparticles present. However, those skilled in the art will appreciate that electrostatic repulsion may also promote stabilization of the polymer material. Such electrostatic forces have little stabilizing effect in liquids with relatively high ionic strength. It will be understood that the copolymers used in accordance with the present invention do not result in The steric stabilizing function of the steric stabilizer is to maintain the nanoparticle material in a dispersed state in such a liquid. They play an important role in enabling the body to maintain a stable or stable state.

[0106] The copolymer steric stabilizers used in accordance with the present invention are useful for preventing the formation of nanoparticles in an in vivo fluid environment. It has been found to be particularly effective in promoting dispersion of the material.

[0107] As used herein, terms such as "polymer" or "polymer segment" mean , is intended to be a reference to a polymer chain resulting from the polymerization of monomers. The polymer component or polymer segment may comprise or consist of polymerized monomer residue units. Such polymer components or polymer segments may be any suitable polymer. In one embodiment, the polymers described herein (e.g., (fixing, steric stabilization and coupling) polymer segments are They are prepared by polymerization of monomers. The polymer chains have non-polymeric chains covalently bonded to them. It may have a moiety, such as a mapping group or a light-emitting group (and may be part of the polymer chain) (having covalently bonded non-polymeric components). As used herein, "copolymer" refers to a The term "polymer chain" refers to a polymer chain containing two polymer segments of different composition. It is intended to

[0108] The copolymer steric stabilizers used in accordance with the present invention comprise a steric stabilizing polymer segment include.

[0109] Those skilled in the art will appreciate the nature of the monomers that can be polymerized to form such polymers. , will appreciate the variety of polymers that can be used as the steric stabilizing polymer segment. The steric stabilizing polymer segment is polyacrylamide (PA), polyvinyl alcohol (PVA), Polyvinyl alcohol (PVA), polyalkylene oxides (e.g., polyethylene oxide (PEO), poly Polypropylene oxide (PPO), polyoxamer, polyhydroxyethyl acrylate Poly-N-isopropylacrylamide, Polydimethylamino-ethyl methacrylate Polyvinylpyrrolidone (PVP), polyacrylic acid (PAA), polymethacrylic acid Polyvinyl esters, polyvinyl amides, polysulfonated divinylbenzenes, poly- L-lysine, polyaspartate, polylactic acid, polyethyleneimine, polyalkylcyano acrylate, polyaspartate, polymaleic anhydride, polymaleic acid, or It may comprise or consist of two or more of the copolymers described above. Suitable monomers that can be used to form the steric stabilizing polymer segment include are acrylamide, vinyl alcohol, alkylene oxide (e.g., ethylene oxide) propylene oxide), hydroxyethyl acrylate, N-isopropyl acrylate Amide, Dimethylamino-ethyl methacrylate, Vinylpyrrolidone, Acrylic acid, Meth Acrylamide, vinyl ester, vinyl amide, sulfonated divinylbenzene, L-lysine amine, aspartate, lactic acid, ethyleneimine, alkyl cyanoacrylate, aspartate Examples include maleic anhydride, maleic acid, or copolymers of two or more of the above. , but not limited to these.

[0110] When the steric stabilizing polymer segment comprises a polyalkylene oxide, the polyalkylene Oxides include polyethylene glycol, polypropylene glycol and their derivatives. The polyalkylene oxide polymer may be end-capped with an alkyl group. The alkyl group may be an alkyl group having 1 to 6 carbon atoms, such as a methyl group, an ethyl group, It may be a propyl group or an isopropyl group.

[0111] Considering that the steric stabilizing polymer segment forms only part of the copolymer steric stabilizer, Considering this, rather than defining the steric stabilizing polymer segment in terms of its number average molecular weight, Instead, it is better to refer to the number of polymerized monomer units that collectively form the segment. It may be more useful to use polymers that can collectively form steric stabilizing polymer segments. Although there is no particular limit to the number of such units, in some embodiments of the present invention, the steric stabilized polymer - The segment has less than 70 polymerized monomer residue units that constitute the entire polymer segment It may be desirable to include, and in certain embodiments, the entire polymer segment The polymerized monomer residue units constituting the polymerized monomer residue units are about 40 to about 60, for example, There are about 50 of them.

[0112] In some embodiments, the steric stabilizing polymer segment comprises polymerized monomer residue units. Contains approximately 10 to 70 pieces.

[0113] The steric stabilising polymer segment may be a homopolymer or a copolymer.

[0114] In one embodiment, the steric stabilizing polymer segment is a polyacrylamide-copolymer. The block copolymer comprises or consists of a alkylene oxide block copolymer. The block copolymers have from about 8 to about 60 polymerized acrylamide units and from about 2 to about 10 It may comprise or consist of polymerized alkylene oxide units.

[0115] In another embodiment, the steric stabilizing polymer segment is a polymerized alkylene oxide monomer. Contains approximately 10 to 13 digits.

[0116] Those skilled in the art will appreciate that polymerized alkylene oxide units give rise to polyalkylene oxides. You will understand that.

[0117] Polymeric steric stabilizers, polymer mapping moieties and polymers used in accordance with the present invention Each of the light-emitting moieties comprises an immobilized polymer segment.

[0118] "Anchoring polymer segment" refers to a polymer chain that is attached to the surface of the nanoparticle material. The nanoparticle material has an affinity for and is designed to immobilize a given entity via one or more binding groups. A given polymer entity (i.e., polymer steric stabilizer, polymer mapping) functions as follows: The term "polymeric light-emitting moiety" refers to a segment or region of the polymeric light-emitting moiety (e.g., polymeric light-emitting moiety and polymeric light-emitting moiety). may form part of the polymer chain backbone or may exist as a side chain from the polymer chain backbone. The binding group may be any element or group that has a binding affinity for the nanoparticle material. For example, the binding group may be any molecule that has a binding affinity for iron or iron oxide. Suitable linking groups that can be used include one or more ribonucleotides or molecules. A group containing a phosphorus (P) atom, a group containing one or more oxygen (O) atoms, or one or more sulfur (S) atoms groups containing one or more nitrogen (N) atoms, and groups containing two or more of the above atoms Examples of groups include:

[0119] In one embodiment, the anchoring polymer segment may comprise a phosphate group, a phosphonate group, a dimer group, or a DMSA group, sulfate group, sulfonate group, catechol group, carboxylate The polymer comprises one or more binding groups selected from groups, amine groups and silane groups.

[0120] By being a polymer segment, the immobilized polymer segment is a polymerized monomer residue. In particular, the segments may be formed in accordance with the requirements for nanoparticle materials. The immobilized polymer segment comprises polymerized monomer residues that generate the binding affinity. The polymerized monomer residues may be the same or different.

[0121] Immobilized polymer segments that provide multiple sites for binding interactions with nanoparticle materials. The ability of the copolymer to provide the excellent stabilization properties provided by the copolymer steric stabilizer is at least partially responsible for the excellent stabilization properties provided by the copolymer steric stabilizer. It is thought that this occurs gradually.

[0122] The immobilizing polymer segments each provide a site for binding to a magnetic nanoparticle. At least two polymerized monomer residues, or at least three such polymerized monomer residues or at least 5, or at least 7, or at least 10 All of the polymerized monomer residues that make up the immobilized polymer segment are bound to the nanoparticle material. Although it is not necessary to provide a binding interaction, the immobilizing polymer segment generally Most, if not all, of the polymerized monomer residues that participate in the bonding interactions with the nanoparticle material It is preferable to provide

[0123] Thus, the immobilizing polymer segments collectively immobilize a given entity to the nanoparticle material. Alternatively, it can be described as having multiple binding sites.

[0124] To achieve the desired immobilization effect, the immobilization polymer segment must be oriented relative to the nanoparticle material. The manner in which the immobilizing polymer segment binds to the nanoparticle material is as follows: by electrostatic forces, hydrogen bonds, ionic charges, van der Waals forces or any combination thereof. The particular advantage offered by the immobilized polymer segment is that The advantage of this is that it can provide multiple sites for binding interactions with nanoparticles. Therefore, if a given binding site only produces a relatively weak interaction with the nanoparticle material However, the existence of multiple such sites within a segment makes the nanoparticle material as a whole This allows for secure bonding to the material.

[0125] In one embodiment, the anchoring polymer segment is not covalently attached to the nanoparticle material.

[0126] The anchoring polymer segment required generally depends on the nature of the nanoparticle material to which it is attached. Those skilled in the art will be able to determine the appropriate solids to bond with the surface of a given nanoparticle material. A stabilizing polymer segment could be selected.

[0127] When describing the interaction between the immobilized polymer segment and the nanoparticle material, the segments and It may be convenient to refer to the hydrophilicity and hydrophobicity of the particle material. Generally, if the segment and particle material have similar hydrophilic or hydrophobic properties, suitable For example, when the nanoparticle material wets its surface with an aqueous solution, In the case of a relatively hydrophilic surface, the immobilized polymer segment having hydrophilic properties can be a segment (e.g., in its isolated form, the segment is soluble in aqueous media) Such an example is when the particulate material is applied to its surface. This would be possible if the sensor was of a type that could generate a charge. The segments form a charge to promote ionic bonding between the segments and the particulate material. and polymerized residues of monomers that can be polymerized (e.g., polymerized residues of ionizable monomers). It may be desirable to promote the formation of such charged species. This could be facilitated by adjusting the pH of the liquid carrier present.

[0128] The expression "ionizable monomer" means that the monomer can be ionized in solution. It is meant to include functional groups capable of forming ionic or anionic groups. Such functional groups generally undergo proton loss or acceptance under acidic or basic conditions. Generally, the functional group can be an acidic or basic group (i.e., A group capable of donating or accepting an H atom, respectively. For example, a carboxylic acid function The group is capable of forming a carboxylate anion under basic conditions, and the amine functional group is capable of forming a carboxylate anion under acidic conditions. The functional group can also be used to form a quaternary ammonium cation under ion exchange conditions. Ionization may be possible depending on the process.

[0129] Those skilled in the art will appreciate the nature of the monomers that can be polymerized to form such polymers. , will understand the various polymers that can be used as the anchoring polymer segment. For example, suitable polymers include polyacrylic acid, polymethacrylic acid, polystyrene, polyimide, Taconic acid, poly-p-styrenecarboxylic acid, poly-p-styrenesulfonic acid, polyvinyl Sulfonic acid, polyvinylphosphonic acid, polymonoacryloxyethyl phosphate, polymono Nonacryloxyethylphosphonic acid, Poly-2-(methacryloyloxy)ethyl phospha ester, poly-2-(methacryloyloxy)ethylphosphonic acid, polyethacrylic acid, poly -α-Chloroacrylic acid, polycrotonic acid, polyfumaric acid, polycitraconic acid, polymesa Acetic acid, polymaleic acid, poly-2-(dimethylamino)ethyl and propyl acryl acrylates and methacrylates, and the corresponding poly-3-(diethylamino)ethyl and propyl acrylates and methacrylates, polydimethylaminoethyl methacrylates and Examples of such copolymers include, but are not limited to, those listed in the following. Suitable monomers that can be used to form the mer segment include acrylic acid, methyl acrylate, methyl meth ... Acrylic acid, itaconic acid, p-styrenecarboxylic acid, p-styrenesulfonic acid, vinyls sulfonic acid, vinylphosphonic acid, monoacryloxyethyl phosphate, monoacryloxy Ethylphosphonic acid, 2-(methacryloyloxy)ethyl phosphate, 2-(methacryloyloxy)ethyl phosphate (Iroyloxy)ethylphosphonic acid, ethacrylic acid, α-chloroacrylic acid, crotonic acid, Fumaric acid, citraconic acid, mesaconic acid, maleic acid, 2-(dimethylamino)ethyl and and propyl acrylate and methacrylate, the corresponding 3-(diethylamino)ethyl and propyl acrylate and methacrylate, dimethylaminoethyl methacrylate Examples include, but are not limited to, acetone, acetaminophen ...

[0130] The anchoring polymer segment may contain from about 1 to about 20 phosphonate groups, e.g., 1 phosphonate group. phosphonate group, 2 phosphonate groups, 3 phosphonate groups, 4 phosphonate groups, 5 phosphonate groups, 6 phosphonate groups, 7 phosphonate groups, 8 phosphonate groups phosphonate groups, 9 phosphonate groups or 10 phosphonate groups, 11 phosphonate groups, 12 phosphonate groups, 13 phosphonate groups, 14 phosphonate groups, 15 phosphonate group, 16 phosphonate group, 17 phosphonate group, 18 phosphonate group In some embodiments, the phosphonate group may contain 19 phosphonate groups, 19 phosphonate groups, or 20 phosphonate groups. In some embodiments, the anchoring polymer segment may contain more than 20 phosphonate groups. In certain embodiments, the anchoring polymer segment comprises five phosphonate groups.

[0131] The anchoring polymer segment may be composed of one type of monomer or two or more different types of monomers. Thus, the anchoring polymer segment can be formed by the polymerization of a homopolymer. It may be a polymer segment or a copolymer segment.

[0132] There is no particular limitation on the number of polymerized monomer units that collectively form the immobilized polymer segment. However, in some embodiments of the present invention, the anchoring polymer segment has a relatively low number average molecular weight. It may be desirable to have an amount of the immobilizing polymer segment (the entire segment). less than about 50, or less than about 40, or less than about 30 polymerized monomer residue units It may contain less than 10, or from about 5 to about 25, or from about 5 to about 15.

[0133] In one embodiment, the anchoring polymer segment comprises from 1 to about 30 polymerized monomer residues. Includes place.

[0134] In one embodiment, the anchoring polymer segment comprises one or more ethylenically unsaturated monomers. It is composed of polymerized residues of

[0135] The anchoring polymer segment may be a copolymer steric stabilizer, a copolymer mapping moiety, or sterically stabilized polymer segments or couplings to form copolymer light-emitting moieties. The polymer segments are covalently bonded to either of the polymer segments.

[0136] The anchoring polymer segment may be a steric stabilizing polymer segment or a coupling polymer segment. mer segments, and thereby copolymer steric stabilizer, copolymer The copolymeric nature of the mapping moiety and copolymeric light-emitting moiety is provided.

[0137] The polymer mapping moiety and polymer light-emitting moiety (if used) are coupled The coupling polymer segment comprises a polymer segment. The "coupled" polymer segment is covalently coupled to the means that the immobilized polymer segment is attached to a mapping group or a radiating group, as described herein. This means that the polymer chain is linked or bonded to one of the photoactive groups. These mapping or luminescent groups are generally covalently attached to the coupling polymer segment. The coupled polymer segment also contains a mapping group and and luminescent groups away from the nanoparticle material surface, thereby allowing e.g., mapping groups to be The mapping group and the luminescent group are made available by a receptor on the target site. It plays a role in making it more functional.

[0138] Those skilled in the art will appreciate the nature of the monomers that can be polymerized to form such polymers. It will be appreciated that various polymers may be used as the coupling polymer segment. For example, suitable polymers include polyacrylamide (PA), polyvinyl alcohol, polyvinyl alcohol (PVA), polyalkylene oxides (e.g., polyethylene oxide (PEO) and and polypropylene oxide (PPO), polyoxamers, polyhydroxyethyl acrylates Acrylate, Poly-N-isopropylacrylamide, Polydimethylamino-ethylmethacrylate Polyvinylpyrrolidone (PVP), Polyacrylic acid (PAA), Polymethacrylate amide, polyvinyl ester, polyvinyl amide, polysulfonated divinyl benzene, Poly-L-lysine, polyaspartate, polylactic acid, polyethyleneimine, polyalkyl cyanoacrylate, polyaspartate, polymaleic anhydride, polymaleic acid or Copolymers of any of the foregoing may be included, but are not limited to these. Suitable monomers that can be used to form the spring polymer segment include acrylamide, vinyl alcohol, alkylene oxides (e.g., ethylene oxide and and propylene oxide), hydroxyethyl acrylate, N-isopropyl acrylate amide, dimethylamino-ethyl methacrylate, vinylpyrrolidone, acrylic acid, methacrylate Divinylamide, vinyl ester, vinylamide, sulfonated divinylbenzene, L-lysine , aspartate, lactic acid, ethyleneimine, alkyl cyanoacrylate, aspartate Examples include, but are not limited to, maleic anhydride, maleic acid, and combinations thereof. It will not be done.

[0139] In certain embodiments, the coupling polymer segment comprises the entire polymer segment. In certain embodiments, the polymer core has less than about 100 constituent polymerized monomer residue units. The polymerized monomer residue units constituting the entire fragment are about 30 to about 80, or It has about 50 to about 80 polymerized monomer residue units, for example, about 70 polymerized monomer residue units.

[0140] In one embodiment, the coupling polymer segment comprises polyacrylamide or polyacrylamide.

[0141] In another embodiment, the coupling polymer segment comprises from about 10 to about 100 Contains polymerized monomer residue units or from about 10 to about 100 polymerized monomer residue units It consists of:

[0142] Coupling polymer segments of one or both of the polymer mapping moiety and the light emitting moiety In a further embodiment of the present invention, the coupling polymer segment is a steric stabilizing polymer. - More polymerized monomer residue units than segments e units). For example, the coupling polymer segment may be a steric stabilizing polymer. At least 2, or at least 4, or at least 6, or fewer than the segment at least 8, at least 10, or at least 12, or at least 14, or or at least 16, or at least 18, or at least 20 more polymerized monomers The coupling polymer segment may have a steric stabilizing polymer segment. Approximately 5 to 70 pieces, or approximately 5 to 60 pieces, or approximately 5 to 40 pieces, or or about 5 to about 20, or about 40 to about 70, or about 50 to about 70 more polymerizations It may have monomer residue units.

[0143] Without wishing to be bound by theory, it is believed that the steric stabilizing polymer segments and providing a coupling polymer segment having more polymerized monomer residue units than the This means that protein adsorption and subsequent cellular uptake in macrophages is unlikely. This is thought to play a role in forming a protective surface environment in the tumor microenvironment. It is believed to improve the accumulation of nanoparticle materials.

[0144] Those skilled in the art will be able to determine the appropriate steric stabilizing polymer segment for use with a given nanoparticle material. , an appropriate combination of an immobilizing polymer segment and a coupling polymer segment, Each of these polymer segments is selected to provide the required functionality. It will be possible.

[0145] The copolymer mapping moiety and the copolymer light-emitting moiety each comprise one or more mapping moieties. These mapping groups and luminescent groups each contain one or more Generally, each moiety is covalently coupled to a coupling polymer segment. are.

[0146] One or more of the mapping groups described herein may be fibroblast activation proteins (FAPs). The present invention includes an agent that specifically binds to a target protein.

[0147] "Fibroblast activation proteins" or "FAPs" are activated by various hormones and extracellular It is a cell surface-expressed protease that acts on matrix components. Structurally, FA P has a 6 amino acid cytoplasmic tail, a 20 amino acid single transmembrane domain, and It consists of 734 extracellular domains.

[0148] FAP is expressed during development and is very rare in healthy adult tissues. It is highly upregulated in a variety of cancers and in cells of the tumor microenvironment.

[0149] Thus, one or more mapping groups containing agents that specifically bind to the FAP are and selectively binding to tumor-associated stromal cells in a subject upon administration of the nanoparticles. Suitable mapping groups include fibroblast activation proteins, which target FAPs. Inhibitors, peptides, proteins and antibodies are listed.

[0150] In one embodiment, the agent is selected from the group consisting of small molecule inhibitors and antibodies or antigen-binding fragments thereof. fragments.

[0151] In one embodiment, the agent is a small molecule inhibitor. In another embodiment, the agent is a FAP inhibitor. It is a harmful agent.

[0152] Examples of suitable FAP inhibitors include those having the following structures: [ka]

[0153] In the formula, R 1 and R 2 are the same or different and each independently represent hydrogen, halo, is selected from the group consisting of alkyl and alkyl having 1 to 4 carbon atoms;

[0154] R 3 is an alkyl having i to 4 carbon atoms, a nitrile, or an isonitrile,

[0155] R 4 , R 5 and R 6 are the same or different and each independently represent hydrogen, halo, The alkyl group is selected from the group consisting of alkylene and alkyl having 1 to 4 carbon atoms.

[0156] In the formula, R 1 and R 2 are each halogens.

[0157] In the formula, R 1 and R 2 Each of is fluorine.

[0158] In the formula, R 3 is a nitrile.

[0159] In the formula, R 4 , R 5 and R 6 Each of is hydrogen.

[0160] In one embodiment, the agent is an antibody or an antigen-binding fragment thereof.

[0161] Other suitable examples of FAP inhibitors include those described in WO 2013107820. U.S. Patent Application Publication No. 20200330624, European Patent Application Publication No. 3763 726 and U.S. Pat. No. 7,399,869. However, the present invention is not limited to these.

[0162] As used herein, the term "antibody" refers to an antibody that specifically binds to a target antigen or Any antibody containing at least one complementarity-determining region (CDR) that specifically interacts with a target antigen. The term "antibody" is understood to mean any antigen-binding molecule or molecular complex. It contains two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. The heavy chains comprise full-length immunoglobulin molecules containing the heavy chains of: Heavy chain variable region (HCVR, VH or V H and the heavy chain constant region The heavy chain constant region typically contains three domains -C H 1. C H 2 and C H Includes 3. Each light The chains are composed of light chain variable regions (LCVR, VL, VK, V K or V L (sometimes abbreviated as) The light chain constant region usually consists of one domain (C L 1) V H and V L The region is further interspersed with more conserved regions, also called framework regions (FRs). Each V can be subdivided into hypervariable regions called complementarity-determining regions (CDRs), which are present in each V. H and V L are usually in the following order from amino terminus to carboxy terminus: Three Cs arranged in FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 Includes DR and four FRs.

[0163] Also, as used herein, "immunoglobulin" (Ig) is used herein to refer to a Class IgG, IgM, IgE, IgA, or IgD (or any subclass thereof) It is defined as a protein belonging to the group, and includes all known antibodies and their functional fragments. A "functional fragment" of an antibody / immunoglobulin retains the antigen-binding region. Defined as a fragment of an antibody / immunoglobulin (e.g., the variable region of an IgG).

[0164] An "antigen-binding region" or "antigen-binding fragment" of an antibody typically refers to one or more found in the hypervariable regions, i.e., CDR-1, CDR-2 and / or CDR-3 regions However, the variable "framework" regions also serve to provide a scaffold for the CDRs. These fragments can play an important role in antigen binding. " includes F(ab')2 fragments, Fab fragments, scFv, or single immunization globulin variable domain or single domain antibody polypeptides, e.g., single heavy chain variable domains The domains of the constructs containing the main or single light chain variable domains are included. F(ab') 2 or Fab, C H1 Domain and C L Intermolecular disulfide phase formed between domains They may be genetically engineered to minimize or completely eliminate interactions.

[0165] One or more luminescent groups emit electromagnetic radiation or acoustic energy of a desired wavelength after some form of stimulation. The luminescent group can be any chemical entity that releases energy. bioluminescent), electroluminescent, photoluminescent, radioluminescent or thermoluminescent In certain embodiments, the luminescent group emits light of a particular wavelength after absorbing a photon. The photoluminescent group may be fluorescent or phosphorescent.

[0166] In a particular embodiment, the luminescent group is a fluorescent group belonging to the group of cyanine dyes. The base is indocyanine green (ICG; sodium = 4-[2-[(1E,3E ,5E,7Z)-7-[1,1-dimethyl-3-(4-sulfonatobutyl)benzo[e] Indol-2-ylidene]hepta-1,3,5-trienyl]-1,1-dimethylbenzyl zo[e]indol-3-ium-3-yl]butane-1-sulfonate), IR dyes, For example, IRdye800, and sulfocyanine dyes, such as sulfo-Cy3, Examples of suitable dyes include sulfo-Cy5 and sulfo-Cy7. Umiprobe Corporation (Hunt Valley, Maryland, USA) ) is commercially available.

[0167] In one embodiment, the luminescent group is indocyanine green, sulfo-Cy3, sulf It is selected from o-Cy5 and sulfo-Cy7.

[0168] In certain embodiments, the nanoparticle material has at least one copolymer steric stabilizer on its surface. The targeting agent and at least one copolymer mapping moiety are attached. In the present invention, the nanoparticle material has at least one copolymer steric stabilizer on its surface, at least The copolymer has attached thereto one copolymer targeting moiety and at least one copolymer light-emitting moiety. do.

[0169] As contemplated herein, nanoparticle materials or compositions comprising nanoparticle materials may be used in combination with other nanoparticle materials. The present invention can be used for diagnostic and / or therapeutic applications.

[0170] Thus, in one embodiment, a. administering to a subject a nanoparticle material or composition according to the present invention; b. detecting the nanoparticle material; a method for mapping tumor margins in a subject, the method comprising: A method is provided for accumulating tumors in a microenvironment, thereby mapping tumor margins.

[0171] As used herein, the term "mapping" or "tumor mapping" refers to the mapping of cancer Generally, tumor mapping is a procedure to establish the location and extent of a tumor. This is done before cancer treatment is administered.

[0172] As used herein, the terms "tumor" and "cancer" refer to tumors associated with abnormal cell growth. Such conditions are known to those skilled in the art. In one embodiment, The tumor is a primary tumor. In another embodiment, the tumor is a solid tumor.

[0173] In one embodiment, the cancer is prostate cancer, glioblastoma multiforme, glioma, or pancreatic cancer. , colorectal cancer, breast cancer, head and neck cancer, gastric cancer, esophageal cancer, ovarian cancer, sarcoma and lung cancer In another embodiment, the cancer is prostate cancer.

[0174] In one embodiment, the tumor is mapped in situ. The tumor is mapped prior to tumor tissue resection. Or they can be mapped in situ after tumor tissue resection. In situations where tumor tissue is removed, tumor tissue resection completely removes all tumor tissue from the subject. This has not been confirmed after tumor tissue resection, for example, by conventional pathology. It will be understood that it is not uncommon for nanoparticle materials according to the present invention to be used in the treatment of conventional pathology. Real-time confirmation that all tumor tissue has actually been removed, without the need for testing To achieve this, tumor mapping can advantageously be performed in situ after tumor tissue resection. do.

[0175] In one embodiment, the nanoparticle material is used in ultrasound, x-ray, optical imaging, computed tomography, computed tomography (CT), single photon emission computed tomography (SPECT), positron emission tomography PET, Fluorescence Resonance Energy Transfer (FRET), and Magnetic Resonance Imaging (MRI) ) is detected using an in vivo imaging technique selected from the group consisting of:

[0176] In another embodiment, the method further comprises determining a clinical target volume (CTV) and / or a target volume (CTV) prior to administering the treatment. Or determining the gross target volume (GTV).

[0177] "Gross tumor volume" or "GTV" refers to the location and extent of the gross tumor, i.e., Refers to a tumor mass that can be seen, palpated, or imaged.

[0178] The "clinical target volume" or "CTV" includes the GTV plus margins for subclinical disease spread. It is important to fully treat the CTV to achieve a cure. It is generally accepted in the field.

[0179] Methods for calculating the CTV and GTV are known to those skilled in the art, illustrative examples of which include , including the method described by Burnett et al. (2004, Cancer Im aging,4(2):153-161).

[0180] In another aspect disclosed herein, there is provided a method for treating cancer in a subject in need thereof. 1. A method for a. administering to a subject a nanoparticle material or composition according to the present invention; b. detecting a site of interest where the nanoparticle material accumulates; c. detecting a compound against the cancer at the site of the nanoparticle material detected in step (b). administering an effective amount of a treatment; A method is provided, comprising:

[0181] Treatment regimens for the treatment of cancer can be determined by one skilled in the art and generally include: The type, size, stage and severity of the tumour, as well as the age, weight and general health of the elephant, were assessed. The risk of recurrence depends on factors including, but not limited to, the patient's disease status and receptor status. For example, a person may be at high or low risk of developing recurrent disease. For subjects identified as having a relatively high or developing risk, developing recurrent disease More aggressive treatment compared with subjects considered to be at low or relatively low risk Similarly, for more advanced stages of cancer, e.g., stage For subjects identified with III or IV disease, the less advanced stage A more aggressive treatment regimen may be prescribed for subjects with gliomas compared to subjects with other cancers.

[0182] As used herein, "treat" and "treatment" and the term "treating" refers to any The method also includes treating or providing a condition or symptom of cancer or other undesirable condition. or any other substance that in any way prevents, inhibits, delays, arrests or reverses the onset or progression of a disease or condition. Therefore, terms such as "treating" are used in their most appropriate manner whenever possible. Treatment should also be considered in a broader sense. For example, treatment may involve the patient being kept in a restroom until complete recovery or cure. It does not necessarily mean to treat. In the case of a condition, treatment does not necessarily cure, prevent, inhibit, delay, arrest or reverse all of said symptoms. and / or any combination thereof, including, but not limited to, treating, preventing, inhibiting, delaying, arresting or preventing one or more of the above symptoms. It can be reversed.

[0183] The target of cancer treatment is a human or a disease of economic and / or social importance to humans. mammals of interest, such as non-human carnivores (e.g., cats and dogs), animals (e.g., pigs, adult pigs and wild boars), ruminants (e.g., cattle, oxen, sheep, tigers, animals, e.g., endangered species of cattle, deer, goats, bison and camels), horses, and birds. Birds, including zoo birds and wild birds, more specifically birds that are also vulnerable to human Because of their economic importance, domesticated birds, e.g., turkeys, chickens, The term "subject" is used to refer to any animal, animal species, or fowl, particularly poultry such as ducks, geese, guinea fowl, etc. It does not indicate a specific age and therefore encompasses adult, juvenile and newborn subjects. is intended.

[0184] The terms "subject," "individual," and "patient" are used interchangeably herein and are used interchangeably herein. It refers to any subject to which the disclosure may be applicable. In one embodiment, the subject is a mammal. In another embodiment, the subject is a human.

[0185] As used herein, the term "therapeutically effective amount" refers to a therapeutically effective amount for a subject in need of treatment, especially for cancer. A therapeutic amount or dose sufficient to treat cancer, administered or applied to a mammal, such as a human, The exact amount of treatment to be administered or applied depends on the age, weight, tumor size, etc. of the subject. The doctor can decide based on the size, extent of infection or metastasis, and individual differences in the condition. .

[0186] In one embodiment, the treatment is surgery, radiation therapy, brachytherapy, photodynamic therapy, Photothermal therapy, (cryoablation, focal laser ablation and high frequency ultrasound Focal ablation therapy, chemotherapy, immunotherapy and The combination is selected from the following:

[0187] In one embodiment, the method further comprises determining the CTV and / or GTV before administering the treatment. This includes:

[0188] In another aspect disclosed herein, a. administering to a subject a nanoparticle material or composition according to the present invention; b. detecting nanoparticle material in the subject; A method for diagnosing cancer, comprising: detecting a compound that accumulates in a tissue (such as vascular tissue) of a subject; wherein detection of the nanoparticle material indicates that the subject has cancer.

[0189] In one embodiment, detecting accumulated nanoparticle material in vascular tissue of a subject indicates that the subject has cancer. Indicates that.

[0190] The present invention relates to the administration of medicaments to a subject for use in mapping, diagnostic and / or therapeutic applications. A suitable composition comprises nanoparticles according to the invention dispersed in a pharmacologically acceptable liquid carrier. A composition is provided that includes the child material.

[0191] On a weight percent basis, the nanoparticle materials have a range of copolymer steric stabilizers and and copolymer mapping moieties can be attached. For example, the nanoparticle material can be Copolymer steric stabilizer was applied to the surface at 10% to 90% (by weight) and the copolymer mapping section In certain embodiments, nanoparticles can be bound in an amount of 90% to 10% (by weight). The material was coated on its surface with 10% (by weight) copolymer steric stabilizer and copolymer mappi. 90% (by weight) of the coating moiety, 15% (by weight) of copolymer steric stabilizer and copolymer - 85% (wt.) of the mapping moiety, 20% (wt.) of the copolymer steric stabilizer, and The copolymer mapping portion was 80% (by weight), and the copolymer steric stabilizer was 25% (by weight). ) and 75% (by weight) of the polymer mapping portion, 30% ( 70% (by weight) of the copolymer mapping portion, and 70% (by weight) of the copolymer steric stabilizer 35% (weight ratio) and copolymer mapping portion 65% (weight ratio), copolymer steric Stabilizer 40% (wt.) and polymer mapping moiety 60% (wt.), copolymer - 45% (by weight) steric stabilizer and 55% (by weight) polymer mapping moiety, The polymeric steric stabilizer was 50% (by weight) and the polymer mapping portion was 50% (by weight). ), 55% (by weight) copolymer steric stabilizer and 45% (by weight) polymer mapping moiety. 60% (by weight) of copolymer steric stabilizer and 4% (by weight) of polymer mapping moiety 0% (wt.), copolymer steric stabilizer 65% (wt.) and polymer mapping section 35% (by weight), copolymer steric stabilizer 70% (by weight) and polymer mapper 30% (by weight) of the acrylic resin, 75% (by weight) of the copolymer steric stabilizer, and - 25% (by weight) of the mapping moiety, 80% (by weight) of the copolymer steric stabilizer and The copolymer mapping portion was 20% (by weight), and the copolymer steric stabilizer was 85% (by weight). ) and copolymer mapping moiety at 15% (by weight), or copolymer steric stabilizer 90% (by weight) and 10% (by weight) copolymer mapping moieties can be combined. In certain embodiments, the nanoparticle material has a copolymer steric stabilizer attached to its surface. and 30% (by weight) of the copolymer mapping moiety. can be done.

[0192] Those skilled in the art will appreciate that nanoparticle materials according to the present invention, when dispersed in a liquid carrier, are hydrodynamically stable. It will be understood that the hydrodynamic diameter refers to the nanoparticle material itself and the at least a copolymeric steric stabilizer and a mapping moiety associated or attached to the nanoparticles Therefore, the hydrodynamic diameter of the dispersed nanoparticle material is , the nanoparticle material itself in combination with at least a copolymer steric stabilizer and a mapping moiety. It can be seen that the diameter obtained by the above equation is expressed as follows. In this case, the hydrodynamic diameter is that of the largest hydrodynamic diameter exhibited by the dispersed nanoparticle material. It is thought that this is the case.

[0193] In one embodiment, the hydrodynamic diameter of the dispersed nanoparticle material is less than about 300 nm, less than about 250 nm. less than about 100 nm, less than about 50 nm, less than about 25 nm, or less than about 15 nm be.

[0194] In further embodiments, the hydrodynamic diameter of the dispersed nanoparticle material is approximately 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140 , 150, 160, 170, 180, 190, 200, 210, 220, 230, 240 , 250, 260, 270, 280, 290 or 300 nm.

[0195] For the avoidance of doubt, the term "hydrodynamic diameter" used herein refers to the dispersed nanoparticle material. " refers to dispersed coated nanoparticles (at least about 50% by number) The mean diameter of the nanoparticles is intended to indicate the hydrodynamics of the dispersed coated nanoparticles. The target diameter is determined herein by dynamic light scattering (DLS).

[0196] The nanoparticle materials or compositions according to the present invention can be used in ultrasound, X-ray, optical imaging, computer Computed tomography (CT), single photon emission computed tomography (SPECT), positron PET, Fluorescence Resonance Energy Transfer (FRET), and Nuclear Magnetic Resonance Imaging Use in conjunction with in vivo imaging techniques, including but not limited to magnetic resonance imaging (MRI) This can be done.

[0197] In one application, the nanoparticle material includes a FAP targeting group (e.g., an inhibitor) and Compositions containing the compounds may be used to treat, for example, prostate cancer, glioblastoma, pancreatic cancer, colorectal cancer, breast cancer, and the tumor microenvironment (e.g., tumor-associated stromal cells and This allows for the detection of cells expressing FAP, such as cells within tumor microcirculation. By specifically binding to FAPs expressed by cells in the border, the nanoparticle material Useful for identifying the boundaries and margins of cancer-affected tissue (i.e., tumor mapping) The nanoparticle materials and compositions also can be used to detect (i.e., diagnose) or treat cancer. It is also contemplated herein that nanoparticles may be useful as part of, for example, nanoparticle materials. The development of treatments such as focal therapy, radiotherapy, proton therapy or brachytherapy It can be used for tumor mapping before the start of tumor imaging. Accurate tumor mapping allows for more precise surgical removal of the tumor, Limiting unwanted side effects and achieving suboptimal debulking of the tumor mass This minimizes the risk of bulking.

[0198] Unless otherwise specified, as used herein, the terms "halogen" and "halo" , I, Br, Cl and F.

[0199] In this specification, "alkenyloxyalkyl", "alkylthio", "alkylamino" "Alkyl" used alone or in compound words such as "dialkylamino" and "dialkylamino" The term "alkyl" refers to a linear, branched or cyclic alkyl, preferably an alkyl having 1 to 20 carbon atoms. Examples of straight-chain and branched alkyl are methyl, ethyl, and cycloalkyl. , propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, Amyl, isoamyl, sec-amyl, 1,2-dimethylpropyl, 1,1-dimethylpropyl propyl, hexyl, 4-methylpentyl, 1-methylpentyl, 2-methylpentyl, 3 -Methylpentyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 3,3-dimethyl butyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 1,2,2-trimethyl Propyl, 1,1,2-trimethylpropyl, heptyl, 5-methoxyhexyl, 1-methylpropyl ethylhexyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 4,4-dimethyl pentyl, 1,2-dimethylpentyl, 1,3-dimethylpentyl, 1,4-dimethyl Pentyl, 1,2,3-trimethylbutyl, 1,1,2-trimethylbutyl, 1,1,3 -Trimethylbutyl, octyl, 6-methylheptyl, 1-methylheptyl, 1,1,3 ,3-tetramethylbutyl, nonyl, 1-, 2-, 3-, 4-, 5-, 6- or 7-methylbutyl ethyl-octyl, 1-, 2-, 3-, 4- or 5-ethylheptyl, 1-, 2- or 3-propylhexyl, decyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- and 8- Methylnonyl, 1-, 2-, 3-, 4-, 5- or 6-ethyloctyl, 1-, 2-, 3- or 4-propylheptyl, undecyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-methyldecyl, 1-, 2-, 3-, 4-, 5-, 6- or 7-ethyl 1-, 2-, 3-, 4- or 5-propyloctyl, 1-, 2- or 3-ethyloctyl -butylheptyl, 1-pentylhexyl, dodecyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-methylundecyl, 1-, 2-, 3-, 4-, 5- , 6-, 7- or 8-ethyldecyl, 1-, 2-, 3-, 4-, 5- or 6-propyl nonyl, 1-, 2-, 3- or 4-butyloctyl, 1-2-pentylheptyl, etc. Examples of cyclic alkyl include monocyclic or polycyclic alkyl groups, such as cycloalkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclo Examples include cyclooctyl, cyclononyl, and cyclodecyl. [Example]

[0200] Example 1: Synthesis of magnetic nanoparticles Part (a): Large maghemite particles were produced using the coprecipitation method. In a typical reaction: FeCl₂·4H₂O (20 g) and FeCl₃·6H₂O (27 g) were dissolved in 0.4 M H Cl (500 mL). A 3M solution (500 mL) of iron salt was added to the iron salt solution. The black magnetite nanoparticles formed were magnetically separated. The mixture was washed with Milli-Q water and then 200 ml of iron(III) nitrate (0.34 M in 1 M nitric acid) was added. Magnetite nanoparticles were oxidized to maghemite by heating them at 100 °C for 1 h in mL. The formed brown maghemite nanoparticles were magnetically separated and washed with Milli-Q water. Disperse in water and dialysis using 14,000 kDa molecular weight cutoff dialysis tubing. The particles were then dialyzed in water for 2-3 days to remove impurities. The average diameter was found to be 16.8±3.3 nm.

[0201] Part (b): Coprecipitation method was used to generate fine maghemite particles. In a typical reaction, F eCl2·4H2O (1.46 g) and FeCl3·6H2O (2.7 g) at 0.4 M HCl (50 mL). Ammonia solution (3 M, 50 mL) was added to the iron salt solution using a funnel. The mineral nanoparticles were magnetically separated and washed with Milli-Q water. ) The particles were magnetically red-coated with 20 mL of iron(III) nitrate at 100°C for 1 hour with stirring. The resulting brown maghemite nanoparticles were magnetically separated and washed with Milli-Q water. The particles were dispersed in Milli-Q water and dialysis tubes with a molecular weight cutoff of 14,000 kDa were used. The particles were then dialyzed in Milli-Q water to remove impurities. The particles were found to have an average diameter of 12.4±3.2 nm. The diameter was measured by DLS and found to be 44.5 nm.

[0202] Part (c): Magnetite particles were prepared using the co-precipitation method. In a typical reaction, FeCl2 4H2O (1.46 g) and FeCl3 6H2O (2.7 g) in 2 M HCl (1 The mixture was dissolved in 100 mL of HCl and Milli-Q water (40 mL). A syringe pump was used to add ammonia solution (3 M, 50 mL) to the iron salt solution. The black magnetite nanoparticles were magnetically separated and the pH of the final particle dispersion reached 8.2. The particles were washed five times with Milli-Q water (50 mL) until the particles were completely dissolved. The average diameter was found to be 12.7±3.3 nm.

[0203] Example 2 (Comparative Example): Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid} 5-block-poly(acrylamide) 70 -(Glu-CO-Lys) polymer PS MA targeting moiety and poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 15 -block-(triethylene glycol monomethyl Magnetic nanoparticles (PSMA-targeted nanoparticles) conjugated with a (polymethyl ether) polymeric steric stabilizer Composition of (child) Part (a): Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5-b lock-poly(acrylamide) 70 Synthesis of

[0204] 2-(((butylthio)carbonothioyl)-thio)-propanoic acid (0.5 g), Acrylamido (10.4 g), 4,4'-azobis(4-cyanovaleric acid) (0.050 g) Dioxane (20 g) and water (30 g) were combined in a round-bottom flask. The mixture was purged with nitrogen. After purging with gas, the reaction was carried out at 70°C for 3 hours. The solution was allowed to cool to room temperature and [2-(methacryloyl)-2-methylpropanol] was added. 4,4'-azobis(4-cyclohexyloxy)-ethyl]phosphonic acid (2.0 g) The reaction mixture was purged with nitrogen gas and stirred for 4 hours. The mixture was heated to 0° C. The resulting polymer was precipitated in acetone and recovered by vacuum filtration. The polymer was dissolved in water, precipitated in acetone, and dried in a vacuum oven at 40 °C for 24 h. It was re-purified by

[0205] Part (b): Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5-b lock-poly(acrylamide) 70 Synthesis of -(Glu-CO-Lys)

[0206] Preparation of Glu-CO-Lys. Dissolve 40 mg of Glu-CO-Lys-(t-Bu)3 ester in dichloromethane (DCM) and A concentration of 0 mg / mL was obtained. Nitrogen gas was bubbled through the mixture at room temperature for 2 hours, and then the solvent was removed. The residue was dissolved in 2 mL of 20% aqueous acetic acid, and the mixture was diluted with chloroform. The product was washed three times and concentrated to dryness under high vacuum to give the deprotected Glu-CO-Lys. It was dissolved in acetic acid, lyophilized and stored at 4°C until further use.

[0207] Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid of Glu-CO-Lys }5-block-poly(acrylamide) 70 Conjugation to Part (a) Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid} 5-block-poly (Acrylamide) 70 (250 mg), N-(3-dimethylaminopropyl)-N'- Ethylcarbodiimide hydrochloride (EDC·HCl, 60 mg) and N-hydroxysuccinic acid Dissolve 12 mg of NHS in 8 mL of MES buffer (100 mM, pH 5.5-6.0). The mixture was sonicated in an ultrasonic bath for 10 min and then dissolved in 10 mL of acetone. The NHS-activated polymer was precipitated by centrifugation at 3,000 x g for 5 min. Glu-CO-Lys (20 mg) was dissolved in 10x concentrated PBS buffer, and N The reaction mixture was stirred at room temperature for 20 hours. Lys-conjugated polymers were loaded onto centrifugal filters with 3 kDa molecular weight cutoff membranes. The product was purified using a HPLC column and washed three times with water. The product was diluted to a final concentration of 50 mg / mL and further The mixture was stored at 4°C for further use.

[0208] Part (c): Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid} 5-b lock-poly(acrylamide) 15 -block-(triethylene glycol monomer Synthesis of ethyl ether

[0209] In a round-bottom flask, acrylamide (2.8 g), 4,4'-azobis(4-cyanomethyl) hydroxybenzoic acid) (0.050g), methoxytriethylene glycol modified 2-[[butylsulfa 1.0 g, dioxane (10 g), A solution of 10 g of methylcellulose and 10 g of water was prepared. The mixture was purged with nitrogen gas for 15 minutes, and then The mixture was heated to 70°C with stirring for 2 hours. The mixture was allowed to cool to room temperature and the [2-(methacryloyl) 4,4'-azobis(4-cyano-4-hydroxyethyl)phosphonic acid (2.6 g) The reaction mixture was purged with nitrogen gas for 15 minutes and then stirred for 4 hours. The mixture was heated to 70°C. The resulting polymer was precipitated in acetone and collected by vacuum filtration. The polymer was dissolved in water, precipitated in acetone, and dried in a vacuum oven at 40 °C for 24 h. The polymer was re-purified by drying. The chemical structure of this polymer is shown in Figure 2.

[0210] Part (d): Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5-b lock-poly(acrylamide) 70 -(Glu-CO-Lys) 30% and poly{[2 -(methacryloyloxy)-ethyl]phosphonic acid}5-block-poly(acrylic acid Mid) 15 -block- (triethylene glycol monomethyl ether) 70% mixture Particle stabilization using compounds

[0211] Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid prepared in part (c) }5-block-poly(acrylamide) 15 -block-(triethylene glycol) monomethyl ether) (36 mg) and part (b) poly{[2-(methacryloyl {(hydroxy)-ethyl]phosphonic acid}5-block-poly(acrylamide) 70 -(G Lu-CO-Lys (40 mg) was dissolved in 2 mL of water. The pH was adjusted to 4.

[0212] of magnetic particles (7 wt% solids, 1 g) was added to the polymer mixture during ultrasonic treatment. The pH was adjusted to 5.5 after 10 minutes and then to 7.0 after an additional 10 minutes of sonication. After the ultrasonic treatment was continued for a total of 30 minutes, the mixture was transferred to a membrane with a molecular weight cutoff of 100 kDa. Unbound polymer was removed using a centrifugal filter. The coated nanoparticles were then washed with water. The solution was washed three times with PBS and diluted with saline to obtain an isotonic dispersion of 30 mg Fe / mL.

[0213] Dispersed nanoparticles in 0.9% saline solution as a suspending medium were measured by DLS. The z-average was 64.2 nm. The particle size intensity distribution is shown in Figure 3.

[0214] Example 3: Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid} 5-blo ck-poly(acrylamide) 70 -FAPI copolymer mapping moiety and poly{[ 5-block-poly(acrylic acid) amide) 15 -block-(triethylene glycol monomethyl ether) copolymer Synthesis of steric stabilizer-conjugated magnetic nanoparticles (FAP mapping nanoparticles). Part (a): Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5-b lock-poly(acrylamide) 70 -FAPI synthesis.

[0215] Example 2 - Part (a) Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid Acid}5-block-poly(acrylamide) 70 (50 mg), EDC·HCl (12 mg) and NHS (3 mg) in 2 ml of MES buffer (100 mM, pH 5.5-6.0). The solution was sonicated in an ultrasonic bath for 10 min, and then dissolved in 6 mL of acetone. The S-activated polymer was precipitated. The precipitate was collected by centrifugation at 3000 x g for 5 minutes. FAPI (3 mg) was dissolved in DMSO (50 μL) and diluted with 10x PBS buffer. This was further diluted to a total volume of 2 mL with HCl and then added to the NHS-activated polyacrylamide polymer. The reaction mixture was stirred at room temperature for 20 hours. The FAPI conjugated polymer was The product was purified using a centrifugal filter with a molecular weight cutoff membrane of a and washed three times with water. The product was diluted to a final concentration of 50 mg / mL and stored at 4°C for further use. The chemical structure of the polymer is shown in Figure 4.

[0216] Part (b): Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5-b lock-poly(acrylamide) 70 -FAPI copolymer mapping portion 30% and 5-block-poly(trimethylsilyl)-2-(2-methyl-2-propanol)-2-one acrylamide) 15 -block-(triethylene glycol monomethyl ether) copolymer Particle stabilization using a mixture with 70% polymer steric stabilizer

[0217] Example 2 - Part (c) Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid Acid}5-block-poly(acrylamide) 15 -block-(triethyleneglycol) 18 mg of poly[[2-(methacrylamide monomethyl ether]] prepared in part (a) {(trimethyloloxy)-ethyl]phosphonic acid}5-block-poly(acrylamide) 70 -FAPI (20 mg) was dissolved in 2 mL of water. The pH was adjusted using NaOH (0.1 M). The polymer mixture was mixed with the magnetic particles (35 mg) of Example 1 part (a) using ultrasonic waves. After 10 minutes, the pH was adjusted to 5.5, followed by an additional 10 minutes of ultrasonic treatment. After the ultrasonic treatment, the pH was adjusted to 7.0. After the ultrasonic treatment was continued for a total of 30 minutes, the pH was adjusted to 7.0. Unbound polymer was removed using a centrifugal filter with a molecular weight cut-off membrane. The coated nanoparticles were washed three times with water and diluted with saline to a concentration of 30 mg Fe / mL. An isotonic dispersion was obtained.

[0218] Dispersed nanoparticles in 0.9% saline solution as a suspending medium were measured by DLS. The z-average was 59.7 nm. The particle size intensity distribution is shown in Figure 5.

[0219] Part (c): Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid} 5-b lock-poly(acrylamide) 70 Copolymer with 30% non-targeting moiety and poly{[2-( {methacryloyloxy)-ethyl]phosphonic acid}5-block-poly(acrylamide) ) 15 -block-(triethylene glycol monomethyl ether) copolymer Non-targeted particles (non-targeted nanoparticles) were used as a control, using a mixture with 70% of the targeting agent. particle stabilization.

[0220] Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid of Example 2 part (c) }5-block-poly(acrylamide) 15 -block-(triethylene glycol) 18 mg of poly{[2-(methacryloyloxymethyl ether)] of Example 2 part (a) {(2-hydroxy-ethyl)phosphonic acid}5-block-poly(acrylamide) 70 (2 0 mg) was dissolved in 2 mL of water. The pH was adjusted to 4 with NaOH (0.1 M). The polymer mixture was added to the magnetic particles (35 mg) of Example 1 part (a) while sonicating. After 10 minutes, the pH was adjusted to 5.5, then sonicated for another 10 minutes before being adjusted to 7. After sonication was continued for a total of 30 minutes, the molecular weight cutoff was adjusted to 100 kDa. Unbound polymer was removed using a centrifugal filter with a membrane. The nanoparticles were washed three times with water and diluted with saline to obtain an isotonic dispersion of 30 mg Fe / mL. Ta.

[0221] Example 4: Animal studies using tracers for prostate tumor delineation Six to eight week old male NODscid gamma mice were inoculated with human prostate cancer cells (LNCaP). was injected directly into the prostate. After 4 to 6 weeks of tumor growth, the mice were (d) and (b) of Example 3, part (d) and (c) of Example 3, part (b), respectively. magnetic nanoparticles bearing a targeting moiety, or a targeting moiety (prepared in Example 3-part (c)) The unsaturated magnetic nanoparticles were injected into the tail vein at 15 mg / ml at 40 mg / kg. After 24 hours, the mice were sacrificed and tissues were collected for analysis. Fixed in 0% neutral buffered formalin and plated on 1% agar containing 2 mM gadopentetate dimeglumine. The specimens were mounted in gel and subjected to T2-weighted MRI using a Siemens 3.0T scanner (Figure 1). 6) Analysis of the mean signal intensity of the tumor revealed that the nanoparticles had a higher signal intensity than those without a mapping moiety. In comparison, FAP mapping nanoparticles were shown to increase contrast by 71%. PSMA-targeted nanoparticles provide contrast compared to nanoparticles without a targeting moiety. It only increased by 18%.

[0222] Resected prostate tumors fixed in 10% neutral buffered formalin were mounted on paraffin blocks. Sections of 5 μm thickness were cut and stained with Prussian blue (Fig. 7, 5x magnification). The presence of iron nanoparticles was visualized (indicated by dark blue staining in Fig. 1). PSMA-targeted nanoparticles Alternatively, mice injected with FAP mapping nanoparticles showed increased staining for iron, suggesting that Mice injected with nanoparticles containing no targeting / targeting moiety showed minimal staining. In contrast, visual inspection showed that tumors in mice injected with FAP mapping particles were iron-rich. This showed increased staining, in contrast to PSMA-targeted nanoparticles. It was observed that the particles were taken up especially near the tumor border and along the vasculature.

[0223] After a 24-hour nanoparticle uptake period, mice were injected intraperitoneally with pentobarbital. The mice were fixed in 10% neutral buffered formalin and transcardially perfused. Whole fixed mice were purchased from Bruker. The patients were examined by T2-weighted MRI scan using a 14.1T system (Fig. 6). The tumors in mice injected with PSMA-based nanoparticles showed increased activity in prostate tumors, particularly in the peritumoral area. The tumor showed increased low signal intensity / negative contrast in the 3D image (Figure 8, tumor indicated by white arrow).

[0224] Example 5: Synthesis of short poly{ [2-(methacryloyloxy)-ethyl]phosphonic acid}5-block-poly(acryloyloxy)-ethyl amide) 30 -FAPI copolymer mapping portion and short poly{[2-(methacryloyl)- {(2-methyl-2-methyl-1,2-dihydro-2 ... 10 - Block-(triethylene glycol monomethyl ether) copolymer steric stabilizer Synthesis of combined maghemite nanoparticles (small maghemite FAP mapping nanoparticles). Part (a): Short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 30 Synthesis of

[0225] 2-(((butylthio)carbonothioyl)-thio)-propanoic acid (0.2 g), arylamide (1.8 g), 4,4'-azobis(4-cyanovaleric acid) (0.020 g), Dioxane (3.6 mL) and water (4 mL) were combined in a round-bottom flask. The mixture was heated under nitrogen. After purging with nitrogen gas, the reaction was carried out at 70°C for 2.5 hours. The solution was allowed to cool to room temperature and [2-( methacryloyloxy)-ethyl]phosphonic acid (0.489 g) and 4,4'-azobi 4-cyanovaleric acid (0.020 g) was added, and the reaction mixture was purged with nitrogen gas. The resulting polymer was precipitated in acetone and centrifuged. The polymer was dissolved in water, precipitated in acetone, and dried under vacuum for 48 hours. The mixture was re-purified by

[0226] Part (b): Short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 30 -FAPI synthesis

[0227] Part (a) Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid} 5-b lock-poly(acrylamide) 30 (150 mg), EDC·HCl (72 mg) and Dissolve 18 mg of HCl and NHS in 8 mL of MES buffer (100 mM, pH 5.5-6.0). The solution was sonicated in an ultrasonic bath for 10 min, and then filtered with a 1 kDa molecular weight cutoff membrane. The activation solution was removed using a centrifugal filter with DMSO. The solution was dissolved in 50 μL of HCl and further diluted with 10x PBS buffer to a total volume of 8 mL. The reaction mixture was then added to the NHS-activated polyacrylamide polymer. The reaction mixture was then incubated at room temperature for 20 hours. The short FAPI conjugated polymer was passed through a membrane with a molecular weight cutoff of 1 kDa. The product was purified using a centrifugal filter and washed three times with water. The polymer was diluted to 0.5 mL and stored at 4 °C for further use. The chemical structure of this polymer is shown in Figure 9. show.

[0228] Part (c): Short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 10 -block-(triethylene glycol monomethyl Synthesis of methyl ether

[0229] In a round-bottom flask, acrylamide (0.93 g), 4,4'-azobis(4-cyano) Valeric acid) (0.030g), methoxytriethylene glycol modified 2-{[butylsulf Thio[phenyl]carbono[thioyl]sulfanyl]propanoic acid (0.5g), dioxane (7m A solution of 1 mL of HCl (100%) and 4 mL of water was prepared. The mixture was purged with nitrogen gas for 15 minutes, and then The mixture was heated to 70°C with stirring for 1.5 hours. The mixture was allowed to cool to room temperature and the [2-(methacryloyl) 4,4'-azobis(4-cyano)-1,4'-diamino-2,4'-diamino ...2,4'-diamino-1,4'-diamino-2,4'-diamino-2,4'-diamino Valeric acid (0.030 g) was added. The reaction mixture was purged with nitrogen gas for 15 minutes and then stirred for 2 hours. The mixture was heated to 70°C for 1 hour. The resulting polymer was precipitated in acetone and recovered by centrifugation. The polymer was dried in vacuum for 48 hours. The chemical structure of this polymer is shown in Figure 10. Shown below.

[0230] Part (d): Short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 30 -FAPI copolymer mapping portion 30% and short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5-block- Poly(acrylamide) 10 -block-(triethylene glycol monomethyl ether Maghemite nanoparticles (Tracer 1, Stabilization of 30% FAP mapping maghemite nanoparticles

[0231] Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid prepared in part (c) }5-block-poly(acrylamide) 10 -block-(triethylene glycol) 44 mg of poly[2-(methacryloyl methyl ether)] prepared in part (b) {(2-methyl-2-methyl-1,2-dihydro-2 ... 30 - FAPI (50 mg) was dissolved in 2 mL of water. The pH was adjusted to 4 with NaOH (0.1 M). The polymer mixture was adjusted to the following: Example 1 part (b) (30 mg / mL) magnetic particles in water After 10 minutes, the pH was adjusted to 5.5, and then 60 mg of ethanol was added to the 100 ml of ethanol solution. After 10 minutes of sonication, the pH was adjusted to 7.0. After a total of 30 minutes of sonication, Unbound polymer was removed using a centrifugal filter with a 10 kDa molecular weight cutoff membrane. The coated nanoparticles (Tracer 1) were washed three times with water and diluted with saline. This gave an isotonic dispersion of 20 mg Fe / mL.

[0232] Part (e): Short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 30 -FAPI polymer mapping part 100% Maghemite nanoparticles (Tracer 2, 100% FAP mapping) ) stabilization.

[0233] Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid prepared in part (c) }5-block-poly(acrylamide) 30 -FAPI (50 mg) with water to a total volume of 2 The polymer was diluted to 1 mL. The pH was measured and found to be 6.9. b) (30 mg / mL) was added to 30 mg of magnetic particles in water while ultrasonic treatment was performed. After the treatment, the pH was measured and found to be 5.8. The pH was adjusted to 7.0 while ultrasonic treatment was continued for a total of 20 minutes. Remove unbound polymer using a centrifugal filter with a 0 kDa molecular weight cutoff membrane The coated nanoparticles (Tracer 2) were washed three times with water and diluted with saline. As a result, an isotonic dispersion of 20 mg Fe / mL was obtained.

[0234] Part (f): Short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 30 Copolymer with 30% non-targeting moiety and short poly{ [2-(methacryloyloxy)-ethyl]phosphonic acid}5-block-poly(acryloyloxy)-ethyl amide) 10 -block-(triethylene glycol monomethyl ether) copolymer - Non-targeted maghemite nanoparticles used as a control in a mixture with 70% steric stabilizer (Tracer 3, non-targeted maghemite nanoparticles) stabilization.

[0235] Poly{[2-(methacryloyloxy)-ethyl]phosphonic acid prepared in part (c) }5-block-poly(acrylamide) 10 -block-(triethylene glycol) 44 mg of poly[2-(methacryloyl monomethyl ether)] prepared in part (a) {(2-methyl-2-methyl-1,2-dihydro-2 ... 30 ( 50 mg) was dissolved in 2 mL of water. The pH was adjusted to 4 with NaOH (0.1 M). The polymer mixture was added to 60 mg of the magnetic particles in water (30 mg / mL) from Example 1 part (b). After 10 minutes, the pH was adjusted to 5.5, and then sonicated for a further 10 minutes. After sonication, the pH was adjusted to 7.0. After sonication was continued for a total of 30 minutes, the pH was adjusted to 10 kDa. Unbound polymer was removed using a centrifugal filter with a molecular weight cutoff membrane. The coated nanoparticles (tracer 3) were washed three times with water, diluted with saline, and then An isotonic dispersion of 0 mgFe / mL was obtained.

[0236] Example 6: Microscopic FAP Mapping In Vitro Binding of Maghemite Nanoparticles to FAP test 30% FAP mapping magnetic red in each of parts (d), (e), and (f) of Example 5 Iron ore nanoparticles (Tracer 1), 100% FAP mapping maghemite nanoparticles (Tracer 2) ) and non-targeted maghemite nanoparticles (Tracer 3) against fibroblast activation proteins The binding affinities of the FAP-expressing melanoma cells were compared with their cellular uptake in the FAP-expressing melanoma cell line C32. The cells were evaluated in vitro by comparing the levels of fetal bovine serum in T75 cell culture flasks. in RPMI medium supplemented with 10% FBS and 1% penicillin-streptomycin. C32 cells were cultured and maintained in a 37°C, 5% CO2 incubator. When the solubility reached 70-80%, the cells were passaged. Cellular uptake was measured to determine the binding parent of the nanoparticles. To test for compatibility, C32 cells were plated in 6-well plates at 3 x 10 5 Cell / well density The cells were seeded in the complete cell culture medium described above at 37°C in a 5% CO2 incubator. The sheet was placed and allowed to adhere for 24 hours.

[0237] All nanoparticles were grown in cell culture medium supplemented with only 1% penicillin-streptomycin. underground, 0.150mgFe·mL -1 The growth cell culture medium was prepared at a concentration of The tracer was removed from the sample and replaced with a tracer dispersion. Three replicas were used to test each tracer. C32 cells were incubated with the tracer dispersion in a 37°C, 5% CO2 incubator for 2 h. The cells were incubated for 4 hours. The cells were washed twice with PBS and detached using trypsin. The cell pellet was collected by centrifugation at 500 × g for 5 minutes. The cell pellet was then resuspended in PBS. The plates were washed twice more with HCl and finally dried overnight at 60°C using a heating block. The cell pellet was digested with trace metal grade nitric acid and hydrochloric acid (1:1 by volume), and the sample was The total volume was diluted with water to 3 mL. Iron concentrations were measured by inductively coupled plasma mass spectrometry (ICP-MS). The results, as shown in Figure 11, indicate that the cell Uptake was measured using tracer 1 (30% FAP mapping maghemite nanoparticles) and tracer 2 (1 00% FAP mapping maghemite nanoparticles) than tracer 3 (non-targeted maghemite nanoparticles) particles) were shown to be much less.

[0238] Example 7: Microcopolymer FAP Mapping Magnetic Field in the Presence or Absence of Serum Binding of hematite nanoparticles versus single polymer micro-FAP mapping of maghemite nanoparticles In vitro testing of activity 30% FAP mapping (Tracer 1) in parts (d) and (e) of Example 5, respectively ), 100% FAP mapping (Tracer 2) binding to fibroblast activation protein The combined activity was measured by cellular uptake into the FAP-expressing melanoma cell line C32 using cell growth medium. In vitro by comparing the presence or absence of fetal bovine serum (FBS). C32 cells were cultured as described in Example 6. C32 cells were cultured in 6 wells. 3 x 10 on a plate 5 Cells were seeded in complete cell culture medium at a density of 1000 cells / well at 37°C. The plates were placed in a 25% O incubator and allowed to adhere for 24 hours.

[0239] After 24 hours, the complete cell culture medium was removed from one of the 6-well plates and the penicillin The medium was replaced with one supplemented with 1% lecithin-streptomycin only. Remove the cell culture medium and add 1% penicillin-streptomycin and 10% FBS. The nanoparticles from parts (d) and (e) of Example 5 were added to the medium at 0.150 mgFe mL -1 The C32 cells were incubated with the tracer The mixture was incubated with the dispersion in a 37°C, 5% CO2 incubator for 24 hours. Wash the cells twice with PBS, detach them using trypsin, and centrifuge them at 500 x g for 5 min. The cell pellet was collected by separating the cells from the pellet. The cell pellet was washed twice more with PBS and finally The cells were then dried overnight at 60°C using a heating block. The dried cell pellets were then analyzed for trace metals. The sample was digested with 1:1 volume ratio of nitric acid and hydrochloric acid, and the sample was diluted with water to a total volume of 3 ml. L. Iron concentrations were measured using inductively coupled plasma mass spectrometry (ICP-MS). The collected data was analyzed and significance was tested using an independent sample t-test. The results are shown in Figure 12. From the results, it was found that for both tracers, binding was greater in the presence of FBS than in the absence of FBS. However, the binding of tracer 1 (30% FAP) was significantly reduced. The binding reduction rate of tracer 2 (100% FAP mapping nanoparticles) was 55%. This was statistically significantly smaller than the 63% binding reduction rate of the PEG-1 nanoparticles (p<0.01). No significant difference was observed between tracer 1 and tracer 2 in uptake in the absence of FBS. (p>0.05).

[0240] Example 8: Preparation of short poly{ [2-(methacryloyloxy)-ethyl]phosphonic acid}5-block-poly(acryloyloxy)-ethyl amide) 30 -FAPI copolymer mapping portion and short poly{[2-(methacryloyl)- {(2-methyl-2-methyl-1,2-dihydro-2 ... 10 - Block-(triethylene glycol monomethyl ether) copolymer steric stabilizer Synthesis of combined magnetite nanoparticles (small magnetite FAP mapping nanoparticles). Part (a): Short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 30 -FAPI copolymer mapping portion 30% and short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5-block- Poly(acrylamide) 10 -block-(triethylene glycol monomethyl ether Magnetite nanoparticles (Tracer 4, F) were used in a mixture with 70% of a copolymer steric stabilizer. AP mapping: Stabilization of magnetite nanoparticles (30%).

[0241] Poly{[2-(methacryloyloxy)-ethyl]phosphine prepared in Example 5 part (c) Sulfonic acid}5-block-poly(acrylamide) 10 -block-(triethylene glycol monomethyl ether) and 44 mg of the poly{ [2-(methacryloyloxy)-ethyl]phosphonic acid}5-block-poly(acryloyloxy)-ethyl amide) 30 -FAPI (50 mg) was dissolved in 2 mL of water. NaOH (0.1 M) was added. The pH was adjusted to 4 using the HCl solution of Example 1 during the 10 minute ultrasonic treatment. Part (c) of the polymer mixture was added to 50 mg of magnetic particles in water (25 mg / mL). The pH of the nanoparticles was measured to be 7.8 before adding the The pH was measured to be 5.4. Then, NaOH (0.1 M) was added and the mixture was sonicated. The pH was adjusted to 6.0 and 7.0 while the ultrasonic treatment was continued for a total of 30 minutes. Remove unbound polymer using a centrifugal filter with a 0 kDa molecular weight cutoff membrane The coated nanoparticles (Tracer 4) were washed three times with water and diluted with saline. As a result, an isotonic dispersion of 20 mg Fe / mL was obtained.

[0242] Part (b): Short poly{[2-(methacryloyloxy)-ethyl]phosphonic acid}5 -block-poly(acrylamide) 30 Copolymer with 30% non-targeting moiety and short poly{ [2-(methacryloyloxy)-ethyl]phosphonic acid}5-block-poly(acryloyloxy)-ethyl amide) 10 -block-(triethylene glycol monomethyl ether) copolymer - Magnetite nanoparticles for non-targeted particles used as a control, with a mixture of 70% steric stabilizer. Stabilization of nanoparticles (Tracer 5, non-targeted magnetite nanoparticles).

[0243] Poly{[2-(methacryloyloxy)-ethyl]phosphine prepared in Example 5 part (c) Sulfonic acid}5-block-poly(acrylamide) 10 -block-(triethylene glycol monomethyl ether) and 44 mg of the poly{ [2-(methacryloyloxy)-ethyl]phosphonic acid}5-block-poly(acryloyloxy)-ethyl amide) 30 (50 mg) was dissolved in 2 mL of water. The H was adjusted to 4. The polymer mixture was added to the (30 mg / mL) hydromagnetic powder of Example 1, part (c). Before adding the polymer mixture, 50 mg of nanoparticles were added under ultrasonication. The pH was measured to be 7.8. After the ultrasonic treatment step, the pH was measured to be 5. The pH was then adjusted to 6.0 and 7.4 using NaOH (0.1 M) while treating with ultrasound. The ultrasonic treatment was continued for a total of 30 minutes, and then the molecular weight cutoff was adjusted to 10 kDa. Unbound polymer was removed using a centrifugal filter with an off-coat membrane. The nanoparticles (tracer 5) were washed three times with water and diluted with saline to a concentration of 20 mg Fe / m An isotonic dispersion of L was obtained.

[0244] Example 7: Microscopic FAP Mapping In vitro testing of binding of magnetite nanoparticles to FAP Experiment. Example 8 part (a) for 30% FAP mapping magnetite nanoparticles (Tracer 4) and (b) nontargeted magnetite nanoparticles (tracer 5) to fibroblast activation proteins. The binding affinity of these compounds was evaluated by measuring their uptake in the FAP-expressing melanoma cell line C32. The results were evaluated in vitro by comparing C32 cells as described in Example 6. To measure the cellular uptake and test the binding affinity of the nanoparticles, C32 cells were cultured. 3 x 10 cells in a 6-well plate 5 Seed cells / well in complete cell culture medium as above at a density of The plate was placed in a 37°C, 5% CO2 incubator and allowed to adhere for 24 hours. .

[0245] Add 1% penicillin-streptomycin and 10% FBS to all nanoparticles. 0.150 mg Fe mL in cell culture medium -1 The cell culture medium was prepared at a concentration of The medium was removed from the plate and replaced with the tracer dispersion. Four replicas were used to measure each tracer. C32 cells were incubated with the tracer dispersion at 37°C and 5% CO2. The cells were then incubated in a centrifuge for 24 hours. The cells were washed twice with PBS and then trypsinized. The cells were detached using a syringe and centrifuged at 500 × g for 5 minutes to collect the cell pellet. The plates were washed twice more with PBS and finally dried overnight at 60 °C using a heating block. The dried cell pellet was quenched with trace metal grade nitric acid and hydrochloric acid (1:1 by volume). The iron concentration was determined by inductively coupled plasma mass spectrometry. The results were measured using ICP-MS. As shown in Figure 13, The cellular uptake of tracer 5 (non-targeted magnetite nanoparticles) was significantly higher than that of tracer 4 ( 30% FAP mapping (magnetite nanoparticles) was shown to be much less.

[0246] It will be understood by those skilled in the art that the present disclosure is not limited in its use to the particular applications described. It will be understood that the present disclosure, in its preferred embodiments, is also The present disclosure is not limited with respect to the particular elements and / or features disclosed. The present invention is not limited to any one or more embodiments, but is instead described and defined by the appended claims. Numerous rearrangements, modifications, and substitutions are possible without departing from the scope of the present disclosure. It will be understood.

[0247] Throughout the following specification and claims, unless the context otherwise requires, the words "include" and "include" are used interchangeably. The terms "comprise" and "include" and "includes" "comprising" and variations such as "including" are means the inclusion of a specified integer or group of integers but not the exclusion of any other integer or group of integers. It is understood that:

[0248] Reference to any prior art in this specification is without prejudice to the accuracy or completeness of any information contained in this document, and is hereby incorporated by reference in its entirety. It is not, and should not be construed as, any form of endorsement that would suggest that the It should not be.

[0249] Please note that the claims below are provisional claims only and are not intended to be limiting. This is provided as an example of the range of compounds that may be claimed in future patent applications based on this application. It is not intended to limit the scope of the invention. Therefore, integers may be added or omitted from the exemplary claims at a later date.

Claims

1. A nanoparticle material suitable for administration to a subject, the nanoparticle material having a surface thereof: a copolymer steric stabilizer that promotes dispersion in a liquid, said copolymer steric stabilizer comprising: an anchoring polymer segment having one or more linking groups that attach an agent to the nanoparticle material; and (ii) a steric stabilizing polymer segment different from said anchoring polymer segment. and (b) a copolymer mapping moiety, wherein (i) said an immobilization having one or more linking groups that attach the copolymer mapping moiety to the nanoparticle material; (ii) a polymer segment that specifically binds to fibroblast activation protein (FAP); and (iii) one or more mapping groups comprising an agent that a coupling polymer segment different from the immobilizing polymer segment; a coupling polymer segment attached to one or more mapping groups; - A nanoparticle material having a mapping moiety bonded thereto.

2. 10. The nanoparticle material of claim 1, wherein the surface of the nanoparticle material is provided with (c) a copolymer light-emitting moiety. and (i) having one or more linking groups that attach the polymeric light-emitting moiety to the nanoparticle material. and (ii) visualization of the location of said nanoparticle material in vivo. one or more luminescent groups for emitting light or an acoustic signal in response to light, ii) a coupling polymer segment different from the immobilization polymer segment; a coupling polymer segment that connects the immobilization polymer segment to the one or more luminescent groups; and a copolymer comprising a mer segment and a light-emitting moiety attached to the copolymer.

3. The one or more luminescent groups are indocyanine green, sulfo-Cy3, sulfo 3. The nanoparticle material according to claim 2, wherein the nanoparticle material is selected from sulfo-Cy5 and sulfo-Cy7.

4. The steric stabilizing polymer segment is a polyacrylamide-co-polyalkylene oxide. Polyacrylamide-co-polyalkylene oxide block copolymer 4. The nanoparticle material according to claim 1, comprising:

5. The polyacrylamide-co-polyalkylene oxide block copolymer has about 8 from about 60 polymerized acrylamide units and from about 2 to about 10 polymerized alkylene oxide units The nanoparticle material of claim 4 comprising a unit.

6. 2. The method of claim 1 , wherein the coupling polymer segment is composed of polyacrylamide.

6. A nanoparticle material according to any one of claims 1 to 5.

7. The steric stabilizing polymer segment has 10 to 70 polymerized monomer residue units. The nanoparticle material according to claim 1 .

8. The coupling polymer segment comprises 15 to 100 polymerized monomer residue units.

8. The nanoparticle material according to claim 1, comprising:

9. The coupling polymer segment is more numerous than the steric stabilizing polymer segment. having more polymerized monomer residue units, A nanoparticle material according to any one of claims 1 to 8.

10. 9. The method of claim 1, wherein the nanoparticle material is a magnetic nanoparticle material. Nanoparticle materials.

11. The magnetic nanoparticle material may be iron (Fe), maghemite (γ-Fe ２ O ３ ), magnetite (Fe ３ O ４ 11. The nanoparticle material of claim 10, comprising:

12. The agent that specifically binds to fibroblast activation protein (FAP) is a small molecule inhibitor and an antibody or an antigen-binding fragment thereof. The nanoparticle material according to any one of claims 1 to 4.

13. The nanoparticle material according to any one of claims 1 to 12 is contained in a pharmacologically acceptable liquid carrier. A composition suitable for administration to a subject, including in vivo.

14. 1. A method for mapping tumor margins in a subject, the method comprising: a. The nanoparticle material according to any one of claims 1 to 12 or the nanoparticle material according to claim 13 administering a composition to said subject; b. detecting the nanoparticle material; Including, The nanoparticle material accumulates in the tumor microenvironment, thereby mapping the tumor margin. How to do it.

15. 1. A method for diagnosing cancer, said method comprising: a. The nanoparticle material according to any one of claims 1 to 12 or the nanoparticle material according to claim 13 administering the composition to a subject; b. detecting the nanoparticle material; Including, The detection of accumulated nanoparticle material in tissue of the subject indicates that the subject has cancer. Showing, how.

16. 1. A method for the treatment of cancer in a subject in need thereof, said method comprising: a. The nanoparticle material according to any one of claims 1 to 12 or the nanoparticle material according to claim 13 administering a composition to said subject; b. detecting sites of accumulation of the nanoparticle material in the subject; c. administering to the site of detection of the nanoparticle material in step (b) an effective amount against said cancer. administering a treatment; A method comprising:

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