Boronated nanoscale substrates and uses thereof

Abstract

Various aspects described herein relate to functionalized nanoscale substrates, the functionalized nanoscale substrates comprising functionalized surfaces of the substrates that include boronated moieties.

Description

[Technical Field]

[0001] The present invention relates to boronated nanoscale substrates and uses thereof. [Background technology]

[0002] Nanoscale particles are suitable for many different applications, and the use and effectiveness of these nanoscale particles can be controlled by functionalizing the nanoparticles. [Brief explanation of the drawings]

[0003] [Figure 1A] Transmission electron microscopy (TEM) image of DND nanocrystals. The average crystal size of diamond produced by this process is 4nm-6nm. [Figure 1B] TEM image of synthetic HPHT diamond nanocrystals. Diamond size can vary from less than 5 nm to as large as 500 nm. [Figure 1C] Figure 1 shows a macroscale image of diamond produced by chemical vapor deposition. [Figure 2] A schematic image showing the cubic crystal structure of diamond depicted with all possible nitrogen vacancy sites. The sites are classified based on the orientation of the symmetry axes. Carbon is represented by a solid atom and nitrogen is represented by a shaded atom. [Figure 3] Schematic image illustrating radiochemotherapy using boron-containing agents as irradiation targets: one neutron reacts with a boron-10 nucleus to produce lithium-7 and helium-4. [Figure 4] Schematic of boron nanoparticles encapsulated in liposomal structures and surface functionalized with various ligands. [Figure 5]Fourier transform infrared spectroscopy of organoborane-functionalized NDs. Samples were reacted with various trigonal boron precursors, including BBr, BCl, and BH, for 180 minutes. All samples show a characteristic shift from wavenumber 1105 to wavenumber 1025, indicative of C-B bonding. [Figure 6] Fourier transform infrared spectroscopy of organoborane-functionalized NDs. Samples were reacted with various trigonal boron precursors, including BBr, BCl, and BH, for 24 hours. All samples show a characteristic shift from wavenumber 1105 to wavenumber 1025, indicative of C-B bonding. [Figure 7] Figure 1 shows an SEM image of the unreacted HPHT nanodiamond control. [Figure 8] Images of nanodiamond cores after boron templating, showing the morphology changes following the synthesis protocol using triangular boron precursors. [Figure 9] FIG. 1 shows images showing SEM and EDS data collected on boron carbide. [Figure 10] FIG. 10 shows an EDS scan of an ND-OH sample reacted with BBr3 in DCM. [Figure 11] Graph showing that X-ray absorption spectroscopy shows a core-hole excitation at 289 eV and a second absolute band gap at 305 eV. [Figure 12] Graph showing X-ray photoelectron spectroscopy of organoborane-functionalized NDs. [Figure 13] Graph showing liposome production resulting in polydisperse multilamellar structures. [Figure 14] Graph showing that liposome fabrication using BND resulted in polydisperse multilamellar structures (before purification, analyzed using DLS). [Figure 15] Scanning confocal microscope image at 40x objective of Lucifer Yellow control dissolved in chloroform. [Figure 16] FIG. 1 shows a scanning confocal microscope image at 40x magnification of liposomes with boronated NDOH. DETAILED DESCRIPTION OF THE INVENTION

[0004] Various aspects described herein relate to functionalized nanoscale substrates, which include functionalized surfaces of the substrates that include boronated moieties. The drawings illustrate generally, by way of example, but not by way of limitation, various aspects of the present invention.

[0005] Reference will now be made in detail to several aspects of the disclosed subject matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the illustrated subject matter is not intended to limit the claims to the disclosed subject matter.

[0006] Throughout this specification, values ​​expressed in range format are intended to be interpreted flexibly to include not only the numerical values ​​expressly recited as the limits of the range, but also all individual numerical values ​​or subranges subsumed within that range, as if each numerical value and subrange were expressly recited. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" is interpreted to include not only about 0.1% to about 5%, but also individual values ​​(e.g., 1%, 2%, 3%, and 4%) and subranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the recited range. The statement "about X to Y" has the same meaning as "about X to about Y" unless otherwise indicated. Similarly, the statement "about X, Y, or about Z" has the same meaning as "about X, about Y, or about Z" unless otherwise indicated.

[0007] As used herein, the terms "a," "an," or "the" are used to include one or more than one unless the context clearly dictates otherwise. The term "or" is used to refer to a non-exclusive "or" unless otherwise indicated. The statements "at least one of A and B" or "at least one of A or B" have the same meaning as "A, B, or A and B." Furthermore, it should be understood that phrases or terms used herein and not otherwise defined are for descriptive purposes only and not for limiting purposes. Any use of section headings is intended to aid in the reading and comprehension of the specification and should not be construed as limiting, and information associated with a section heading may be found within or outside of that particular section.

[0008] All publications, patents, and patent documents referred to herein are incorporated herein by reference in their entirety, as if individually incorporated by reference. In the event of a conflict in usage between this specification and those documents so incorporated by reference, the usage in the incorporated references shall be considered supplementary to the usage in this specification, and to the extent of any conflict, the usage in this specification shall control.

[0009] In the methods described herein, unless a temporal or operational order is explicitly recited, acts may be performed in any order without departing from the principles of the invention. Furthermore, specified acts may be performed simultaneously unless express claim language recites that they be performed separately. For example, a claimed act of doing X and a claimed act of doing Y may be performed simultaneously in a single operation, and the resulting process may fall within the literal scope of the claimed process.

[0010] As used herein, the term "about" allows for some variability in values ​​or ranges, for example, within 10%, within 5%, or within 1% of the stated limits of the stated value or range, and includes the exact stated value or range.

[0011] As used herein, the term "substantially" refers to a majority or majority, such as at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%.

[0012] Various aspects of the present disclosure described herein relate to functionalized nanoscale substrates. The substrates include boronated moieties that form functionalized surfaces. Generally, the boronated moieties are 10 B is boron. The substrate is a nanoscale particle or nanoparticle that can comprise diamond, gold, silver, silica, or a mixture thereof. In certain embodiments, the substrate comprises diamond. The major dimension of the nanoscale substrate can range from about 5 nm to about 100 nm, or from about 15 nm to about 80 nm.

[0013] The boronated moieties can be disposed over about 40% to about 100% of the total surface area of ​​the surface of the substrate, or over about 60% to about 100% of the total surface area of ​​the surface of the substrate. The functionalized nanoscale substrates can be incorporated into a delivery vehicle, which can include liposomes that encapsulate the functionalized nanoscale substrates described herein.

[0014] As further described herein, functionalized nanoscale substrates can generally be formed by boronating hydroxylated nanoscale substrates. Boranating the substrate can include reacting the hydroxylated nanoscale substrate with boron tribromide, boron trichloride, or a mixture thereof. Delivery vehicles can be formed by contacting the functionalized nanoscale substrate with one or more lipids, such as phospholipids. The functionalized nanoscale substrate can then be mixed with one or more lipids to form one or more liposomes encapsulating the functionalized nanoscale substrate.

[0015] The functionalized nanoscale substrates can be used to treat tumors. The method of treatment comprises administering a therapeutically effective amount of the functionalized nanoscale substrate to a patient in need thereof. The method further comprises exposing the administered functionalized nanoscale substrate to electromagnetic radiation. Examples of tumors that can be treated include brain tumors. More specifically, tumors that can be treated include cutaneous melanoma or glioblastoma.

[0016] Carboranes and organometallics have shown great promise in the field of medicinal chemistry. Recently, carboranes have been utilized in drug discovery due to their ability to rapidly convert from a trigonal planar configuration (sp2) to a more neutral or tetrahedral configuration (sp3) due to the electron-deficient nature of the carborane complex. More recently, advances in the field of nuclear medicine have shown that carboranes may be useful in boron neutron capture therapy (BNCT). Thermal neutron capture of boron-10 can result in the formation of lithium-7( 7 Li) and alpha particles (helium-4 or 4 Currently, there are several factors that hinder the ability of BNCT to be used safely and effectively. For the treatment to be effective, an adequate number of 10B atoms need to be introduced into the neoplastic cells in an appropriate concentration and only after cellular incorporation of the carborane complex is achieved are they then irradiated.

[0017] Boron-10( 10 Surface modification of hydroxylated fluorescent nanodiamonds with hydroxylated nanoparticles (B) has been proposed for the treatment of cutaneous melanoma, glioblastoma, and other head and neck cancers using neutron capture therapy. To improve the specificity of radiation therapy and the targeted destruction of cancer cells, clinicians 10 The modified oxidized nanodiamonds can be reacted with triangular boron precursors to produce boron-rich surface coatings. These modified diamonds can be 10 It acts as a novel delivery vector for B and can therefore be targeted by thermal neutron capture. It has been reported that cells require an average of 3-7 alpha particles to destroy a cancerous tissue approximately 15 μg in size. 10 Surface functionalization of nanodiamonds with B may allow adequate concentrations to enter the tumorous cells targeted for irradiation, ultimately improving the viability of this therapy. 10 They have yet to be explored as potential substrates for B and could replace the polyhedral boron clusters that have been used in the past as vectors for radioactive boron in cancer therapy. Ultimately, these nanomaterials could potentially address clinicians' need for robust, non-toxic carriers for boron in a variety of medical applications.

[0018] Functionalization of NV nanodiamonds begins with the oxidation of the nanodiamond surface, which improves reactivity by introducing hydroxyl termini at the surface. From here, trigonal planar boron compounds (BH3, BCl3, or BBr3) act as electrophiles and react with tertiary alcohols on the oxidized ND surface. The electron-rich hydroxyl groups on the diamond surface interact directly with the electron-deficient boron compounds to generate a boron shell around the diamond core. This shell is highly reactive, and its size is hypothesized to be controllable by adjusting the amount of time the electrophile reacts with the oxidized diamond or by increasing the concentration of the electrophile used. After purification and removal of by-products, the functionalized samples are analyzed to characterize the degree of boron templating.

[0019] To the desired area 10 In order to improve the localization of B-coated nanodiamonds, liposomes have been proposed as a way to encapsulate functionalized nanodiamonds and safely deliver them directly to tumor sites. Liposomes are essentially amphoteric phospholipid vesicles that form ring-shaped membranes or micelles that can mimic the membranes of cancer cells. Another advantage of using liposomes to deliver boronated diamonds to tumor sites is that liposomes can be further functionalized on the surface to improve their specificity to tumorous cancer cells.

[0020] Diamond samples are analyzed using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), energy-deficient X-ray spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS) to confirm that boronation has indeed occurred at the diamond surface. Liposomes are then fabricated around the functionalized particles and visualized using confocal and other fluorescence microscopy techniques. To improve the feasibility of BNCT, the functionalized nanodiamonds and liposomes are characterized and visualized to confirm the feasibility of these protocols. 10This will help verify that we have successfully generated an effective vector for B delivery.

[0021] Carbon-based nanomaterials are versatile materials favored for their biocompatibility, chemical inertness, and unique thermochemical properties. They have been used in drug delivery, tissue engineering, tissue scaffolding, biosensing, and various other areas of biomedical research. Nanodiamonds (NDs) exist as a tetrahedral carbon lattice, which endows them with highly desirable physical and chemical properties. The carbon within the lattice is arranged in a sp 3 The surface of the tetrahedral structure can be oxidized to produce alcohol-rich surface terminations that can improve reactivity and enable functionalization of the nanoparticles. There are many methods for producing NDs, including chemical vapor deposition, application of high temperature and pressure (HTHP), or detonation. The average size of diamond nanoparticles can range from 5 nm to 100 nm depending on the synthesis or isolation method of the material (Figure 1). Mechanically, NDs have higher toughness and elastic modulus compared to macroscale diamond or other nanoparticles used in biomedical applications, such as gold and silver. At the nanoscale, the sp of NDs 3 Due to their bonding arrangement, their chemical stability and thermal conductivity are similar to those of macroscale diamonds. These material properties ensure that nanodiamond particles are resistant to fracture. Nanodiamond particles also have a high refractive index and are electrically resistive, further enhancing their usefulness as optical sensors or drug delivery vectors. NDs are particularly interesting materials for two reasons: they are small enough to be useful in a variety of biological and nanoscale environments, and they can be selectively modified at the surface to enhance the electrochemical and thermal conductivity properties of NDs.

[0022] Atomic defects, more specifically nitrogen vacancies (NVs), allow scientists to detect small transfers of energy and record energy transfer between molecules in vitro. NV diamonds are naturally fluorescent, absorbing photons in the green range of visible light and emitting detectable signals in the red range. Light is used to selectively excite a single molecule, which then detects the electromagnetic environment (within 1-10 nanometers) by donating a photon to an accepting fluorophore. The energy transfer can then be measured by instruments such as electron-multiplying charge-coupled devices (EMCCDs), which detect the changing fluorescent signal. Advances in the field of microscopy have enabled major breakthroughs in the utility of carbon nanodiamonds as functional biosensors and targets for optically detected magnetic resonance (ODMR). While magnetic resonance can be assessed using many different biophysical techniques, robust ODMR provides useful data for studying single-molecule fluorescence resonance energy transfer (smFRET) events, such as ligand binding or conformational changes in biopolymers, with high specificity. ODMR measurements can be time-dependent, and spin readout can be induced and detected using fluorescence microscopy. When the NV defects are negatively charged and stabilized by the hydroxyl terminations on the surface, it is possible to obtain a readout of the electron spin after stimulation with photons. 1 H nuclear magnetic resonance ( 1 H NMR). Evidence suggests that smFRET can measure nanoscale distances between complexes that exchange radiative energy. Through characterization of modified fluorescent nanodiamonds, we hope to enhance our understanding and optimize the performance of these nanomaterials for biomedical applications and single-molecule experiments.

[0023] Nitrogen-vacancy (NV) defects consist of missing carbon atoms in the diamond lattice structure, directly adjacent to the substituted nitrogen atoms (Figure 2). These NV centers are sensitive to externally applied magnetic fields, making them useful for noninvasively measuring electromagnetic fields in sensitive tissues (such as the brain). The fluorescence signal from NV centers is extremely stable and remarkably resistant to photobleaching or signal intermittency. In recent years, as more data has revealed the importance of NV centers in quantum information processing, substantial efforts have been made to control the charge of NV centers. The negative charge state of NV centers hosts a coherent, long-lived electron spin state in its electrostatic environment, thereby making them useful as electromagnetic probes. To this end, research has been conducted to improve the stability of negative NV charge centers. In the past, researchers have demonstrated that NV centers can be recharged to their ground state (NV) when exposed to a continuous photon source, such as a laser. 0 ) and charge state (NV - To improve the stability of the NV, a potential difference was applied to the diamond surface to induce a spontaneous transition between the NV and the 0 Finally, NDs can be chemically modified to convert NVs into excited states. - The stability of the ion beam can be improved.

[0024] NDs with naturally occurring NV impurities can form complexes on their surfaces with a variety of ligands, including peptides, amines, silanes, and metalloids, and the chemistry of such NDs may be the focus of our research. NDs that retain NV centers offer fluorescent properties and impart special optical properties that can be enhanced or tuned by selective functionalization of the ND surface. Specific modifications can act to enhance the fluorescent signal emitted by the defects, tune colloidal stability, or directly affect the stability of the NV ground state. Boron-containing metalloid modification on the surface of NDs has been explored in an effort to improve the reactivity of diamond surfaces due to their high steric hindrance and chemical inertness. Because organoborane chemistry is characterized by delocalized electron-deficient bonds, boron often aggregates to form three-center, two-electron pair structures. NDs can be oxidized at high temperatures to provide alcohol-rich surfaces. The oxidized nanodiamonds can then be reacted with boron compounds to form metalloid complexes on the diamond surface.

[0025] Boron neutron capture therapy Coating the surface of diamond with boron is useful for performing additional chemistry, but may also hold therapeutic benefits. Boron neutron capture as a radiotherapy has been explored as a possible treatment for aggressive brain tumors since neutrons were discovered in the 1930s. Boron-10 is a material that can be used to capture low-energy neutron beams. 10BNCT has been of particular interest to clinicians due to the highly localized nature of the fission reactions that occur when irradiated with boron (Figure 3). As medical science and pharmaceutical chemistry continued to advance, new boron compounds were synthesized, making BNCT more feasible. By 1968, this therapy was being used successfully in Japan, with 54% of glioblastoma patients in one study surviving more than five years after starting treatment. In the past, first-generation boron agents were few and far between, all of which were chemically unstable, prone to decomposition, or showed poor tumor retention. Direct injection of boron carriers into the tumor site followed by surgical resection and irradiation was only moderately successful. With the advent of third-generation boron compounds, the procedure instead involves intravenous or intrathecal administration of various boron carriers, without the need for immediate surgery, followed by radiation therapy. To minimize the amount of vascular tissue damaged by irradiation, the amount of therapeutic agent in the tumor mass is carefully considered in relation to the amount of agent in the surrounding tissue and blood. Generally, the original boron target is radiolabeled and tracked using positron emission tomography to confirm that it has localized to the tumor. 9 pieces 10 B atoms are required to radiotreat each cell, or approximately 30 mg per gram of tumor mass. 10 It is estimated that B is required. The overall equation governing the principle of BNCT is as follows:

[0026]

number

[0027] When excited by thermal neutrons, the excited 11 The B nuclei decay by fission to produce alpha particles and recoil lithium nuclei along with a small amount of gamma radiation. This radiation has a range of between 5 μm and 10 μm, making it useful for specifically targeting tumors and leaving the surrounding vasculature intact.

[0028] Boron chemoradiotherapy such as BNCT is 10Polyhedral carborane structures are used to deliver B to tumor sites in sufficient quantities. In the past, scientists used first-generation inorganic borates, such as borax and boric acid. These agents were considered unsuitable for radiochemotherapy due to the fact that these compounds were not very specific to tumors and often did not persist after administration. Polyhedral boranes, more specifically icosahedral dicarba-closo-dodecaborane, are relatively nontoxic and pack large amounts of boron into a small volume, making them attractive candidates for radiotherapy. Ideal targets for BNCT are those that can easily cross cell membranes, cross the blood-brain barrier (BBB), and persist intracellularly by evading degradative enzymes. More recently, there has been great interest in using boron-containing compounds such as para-borophenylalanine (BPA) and sodium mercaptoundecahydrododecaborate (BSH). BPA closely resembles the amino acid phenylalanine and the pigment melanin, allowing it to easily penetrate tumor masses. BSH is another polyhedral borate that has been observed to accumulate highly in tumors but not in surrounding tissues. These so-called second-generation boron compounds bioaccumulate and are incorporated into cellular substructures, ultimately becoming robust targets for irradiation and subsequent destruction. With the exception of BPA and BSH, very few boron agents have passed animal testing and clinical trials. The lack of knowledge surrounding boron nanoparticles makes studying the metalloid surface modification of diamond particularly interesting.

[0029] One of the drawbacks of using boron clusters such as decaborane and dodecaborane is that the synthesis of these compounds is often dangerous and expensive. Traditional synthesis of boron clusters or other boron hydrides involves either combining boron trifluoride with sodium borohydride or pyrolysis of diborane gas. This requires specialized equipment for material handling and synthesis on an industrial scale. Generally, the biggest problem is the lack of activated 10The lack of an efficient transport mechanism to carry B compounds from the site of administration to the site of action is further complicated by the diverse subpopulations of cancer cells that may differentially take up boron agents, thereby resulting in the persistence of cancerous tissue. To improve the feasibility of BNCT, alternatives to polyhedral borane clusters must be considered as drug delivery substrates. Nanodiamonds have a number of 10 It is a durable substrate with a high surface-to-volume ratio that can potentially deliver B atoms to their intended targets. To further address the biggest problems facing currently accepted boron drugs or carriers, functionalized nanocrystals can be encapsulated in vesicles or liposomes (Figure 4). Liposomes can safely guide NDs across the BBB, avoiding healthy tissue and evading the immune system, while specifically targeting surface receptors on cancer cells. In this way, boron can be introduced intravenously into a patient's body via NDs and delivered directly to the brain, directly targeting neoplastic cells.

[0030] Liposome Chemistry Liposomes are some of the most successful drug delivery vectors discovered to date. Their fundamental structure is fairly simple, consisting of a simple phospholipid bilayer. This membrane consists of hydrophilic outer and inner walls and a hydrophobic core (Figure 4). One of their greatest uses as biocarriers is their ability to optimize their surface for specific functions. For example, in most drug delivery applications, small amounts of polyethylene glycol (PEG) can be attached to the polar outer surface of liposomes to help them avoid opsonization or phagocytosis. Cholesterol or other steroid molecules can be added to control membrane fluidity and stabilize the hydrophobic core of the liposome. Simple surface modifications can be made to improve tumor specificity. In one study, folic acid was added to the liposome surface because it was found that many tumor cells tend to overexpress folate receptors in the cell membrane. In vitro localization studies showed that folic acid-functionalized liposomes penetrated a BBB model more effectively than liposomes without folic acid coating.

[0031] Liposome production can be approached in many ways, largely dependent on the drug load, the solvent in which the liposomes are dispersed, and the scale of production. Scientists also consider the type of vesicle required for a given application. Unilamellar structures consist of a typical phospholipid shell with an aqueous core, while multilamellar structures consist of one or more concentric spheres. For the purpose of encapsulating boron nanoparticles, unilamellar structures are preferred due to the fact that they retain hydrophilic contents more effectively than multilamellar species. A common approach for preparing small batches of liposomal micelles is via thin film hydration. This technique involves dispersing phospholipids in an organic solvent and rotary evaporating the organic solvent to obtain a lipid thin film. Finally, the thin film is rehydrated in an aqueous buffer. The thin film hydration method often produces many multilamellar liposomes, which require tedious extrusion and sonication to become homogeneous and unilamellar. Reverse phase evaporation and solvent infusion are alternative methods for producing liposomes. These processes work by directly hydrating phospholipids from organic solvents to yield aqueous suspensions containing multilamellar or unilamellar vesicles, depending on the process used (reverse-phase evaporation yields multilamellar species, while solvent infusion yields unilamellar species). Once the ideal liposome preparation method is selected, it is determined in which solvent the drug contents or boron-coated NDs will reside before loading into the liposomal micelles. Lipophilic drugs or highly nonpolar solvents can be mixed with the lipid film prior to hydration. Alternatively, hydrophilic drugs can be added to the aqueous solution used to hydrate the lipid film. Finally, once the liposome species are appropriately labeled and drug-loaded, they can be visualized using different microscopy techniques and further characterized through spectroscopic analysis.

[0032] Fabricating boron nanostructures to better control their delivery for therapeutic applications has helped improve the utility and efficacy of boron neutron capture therapy. Traditionally, pyrolysis of diborane gas using a photon emission source has been used to synthesize boron nanoparticles. A significant drawback of this synthesis method is that laser heating of diborane gas to temperatures above 1000 °C produces amorphous boron nanostructures with a broad size distribution. To reduce the size distribution of boron particles, arc decomposition of diborane gas is preferred because it significantly reduces the number of amorphous structures by producing particles between 55 and 95 nm. Polyhedral boron and carborane clusters have also been fabricated for the purpose of facilitating neutron capture therapy. Transition metal catalysts are used in template-assisted self-assembly to generate block copolymers via pyrolysis of diborane gas. Alternative methods focus on multiphase solution-phase synthesis. Solution-phase synthesis involves wet-milling commercially available boron powder, followed by functionalization with sodium dodecyl sulfate (SDS) or oleic acid to reduce air oxidation and prevent the self-ignition of nanoscale boron particles. To reduce the need for gas-phase decomposition of diborane gas, an air-free method for the synthesis of boron nanoparticles has been proposed. The solution synthesis of organo-capped boron nanoparticles involves the reduction of boron tribromide with sodium naphthalenide in dry dimethoxyethane. This method eliminates the need for the use of flammable and toxic gas-phase diborane. Here, we propose a novel method for the surface functionalization of carbon nanodiamonds by the formation of carborane structures via the reduction of oxidized nanodiamonds with boron tribromide and other reactive boron trihalides.

[0033] When oxidized nanodiamonds are reacted with trigonal boron compounds under inert conditions, the hydroxyl termini can act as nucleophiles to form stable organoborate structures on the diamond surface.

[0034] If boron can be safely templated onto the surface of nanodiamonds, they can be loaded into liposome structures without affecting the surface chemistry of the boron-nanodiamond structures.

[0035] If it is possible to template triangular boron structures onto carbon nanocrystals, it should theoretically be possible to use the same protocol to template boron onto other nanoparticles, such as gold or silica.

[0036] According to various aspects of the present disclosure, the functionalized nanoscale substrate comprises a functionalized surface of the substrate. At least a portion of the functionalized surface is a boronated moiety. The substrate can comprise any suitable material. Examples of suitable materials include diamond, gold, silver, silica, or mixtures thereof. According to various aspects, diamond is a particularly well-suited substrate material. The boronated moiety is: 10 B Contains boron.

[0037] The boronated portion may be from about 40% to about 100% of the total surface area of ​​the surface of the substrate, from about 60% to about 100% of the total surface area of ​​the surface of the substrate, less than about 40%, equal to about 40%, or greater than about 40% of the total surface area of ​​the surface of the substrate, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60% of the total surface area of ​​the surface of the substrate. , 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0038] In some examples, functionalized nanoscale substrates are encapsulated in liposomes. Liposomes have an aqueous solution core in the form of a lipid bilayer surrounded by a hydrophobic membrane, and hydrophilic solutes dissolved in the core cannot easily pass through the bilayer. Hydrophobic chemicals associate with the bilayer. This property can be used to load liposomes with hydrophobic and / or hydrophilic molecules, a process known as encapsulation. Typically, liposomes are prepared in a solution containing the compound to be entrapped. This solution can be either aqueous to encapsulate hydrophilic compounds such as proteins, or in an organic solvent mixed with lipids to encapsulate hydrophobic molecules. Encapsulation techniques can be classified into two types: passive, which rely on stochastic entrapment of molecules during liposome formation, and active, which rely on the presence of charged lipids or a transmembrane ion gradient. An important parameter to consider is the "encapsulation efficiency," which is defined as the amount of compound present in the liposome solution divided by the initial total amount of compound used during preparation. In a more recent development, the application of liposomes in single-molecule experiments has introduced the concept of "single entity encapsulation efficiency," a term that refers to the probability of a particular liposome containing the required number of copies of a compound.

[0039] To deliver molecules to the site of action, lipid bilayers can fuse with other bilayers, such as cell membranes, thus delivering the liposomal contents. However, this is a complex and non-spontaneous event that does not apply to nutrient and drug delivery. By preparing liposomes in a solution of DNA or drugs (which normally cannot diffuse through membranes), they can be delivered (promiscuously) across the lipid bilayer. Liposomes can also be designed to deliver drugs in other ways. Liposomes containing low (or high) pH can be constructed so that dissolved aqueous drugs can be charged in solution (i.e., the pH is outside the drug's pI range). As the pH within the liposome naturally neutralizes (protons can pass through some membranes), the drug is also neutralized, allowing the drug to pass freely through the membrane. These liposomes act to deliver drugs by diffusion rather than by direct cell fusion. However, the effectiveness of this pH-controlled passage depends on the physiochemical properties of the drug in question (e.g., pKa and basic or acidic properties), which for many drugs is very low.

[0040] According to various aspects of the present disclosure, a therapeutically effective amount of the functionalized nanoscale substrate can be administered to a patient in need thereof. The functionalized nanoscale substrate can be encapsulated in liposomes as a pharmaceutical composition or kit. A "therapeutically effective amount" (or "effective amount") of a compound for use in therapy refers to the amount of compound in a formulation that, when administered (to a mammal, such as a human) as part of a desired dosing regimen, alleviates the symptoms, improves the condition, or delays the onset of a disease state in accordance with clinically acceptable criteria for the disease or condition being treated or for cosmetic purposes, e.g., at a reasonable benefit / risk ratio applicable to any medical treatment.

[0041] The term "prophylactic or therapeutic" treatment is art-recognized and includes administration of one or more compounds of the present disclosure to a patient. When administered prior to clinical symptoms of an undesired condition (e.g., a disease or other undesired condition in a host animal), the treatment is prophylactic (e.g., protects the host against acquiring the undesired condition), whereas when administered after the manifestation of the undesired condition, the treatment is therapeutic (e.g., intended to reduce, ameliorate, or stabilize an existing undesired condition or its side effects).

[0042] As used herein, the term "kit" refers to a product (e.g., a drug, kit of parts) that includes the following in one package, or in one or more separate packages: (i) A pharmaceutical composition containing an active pharmaceutical ingredient, one or more additional active pharmaceutical ingredients, and optionally a medical device. The one or more additional active pharmaceutical ingredients may be present in the pharmaceutical composition, i.e., the kit may include one or more packages, where each package contains one pharmaceutical composition containing two or more active pharmaceutical ingredients. The additional active pharmaceutical ingredients may also be present in additional pharmaceutical compositions, i.e., the kit may include separate packages of two or more pharmaceutical compositions, where each pharmaceutical composition contains one active pharmaceutical ingredient.

[0043] or (ii) Pharmaceutical compositions containing active pharmaceutical ingredients and medical devices. A kit may include only one package, or may include one or more separate packages. For example, a kit may be a product (e.g., a medicament) containing two or more vials each containing a defined pharmaceutical composition, where each pharmaceutical composition contains one or more active pharmaceutical ingredients. For example, a kit may include (i.) a vial containing a formed pharmaceutical composition and (ii) additional tablets, capsules, powders, or any other oral dosage form containing one or more additional active pharmaceutical ingredients. The kit may further include a package insert containing instructions on how to administer the pharmaceutical composition and one or more additional active pharmaceutical ingredients.

[0044] As used herein, the term "medical device" means any instrument, apparatus, implant, in vitro reagent, or similar or related article that is used to diagnose, prevent, or treat disease or other conditions and that does not achieve its purpose through pharmacological action in or on the body.

[0045] As used herein, a medical device may be a syringe, an insulin injection system, an insulin infusion system, an insulin pump, or an insulin pen injection device. As used herein, a medical device may be mechanically or electromechanically driven.

[0046] Ingredients in a pharmaceutical composition can be defined as generally recognized as safe ("GRAS"). A complete list of GRAS ingredients can be found in the GRAS Substances (SCOGS) database maintained by the U.S. Food and Drug Administration. About 50% to about 100% of the ingredients in a pharmaceutical composition can be classified as GRAS ingredients; about 75% to about 100%, about 90% to about 100%, less than about 50%, equal to about 50%, or greater than about 50%, 55, 60, 65, 70, 75, 80, 85, 90, 95, or about 100% of the ingredients in a pharmaceutical composition can be classified as GRAS ingredients.

[0047] After administration, the functionalized nanoscale substrate is irradiated with electromagnetic radiation. The functionalized nanoscale substrate is not irradiated with electromagnetic radiation until the substrate is in proximity to a tumor. The tumor can be a brain tumor. Examples of brain tumors include cutaneous melanoma or glioblastoma.

[0048] Example Materials and Methods High pressure, high temperature nanodiamond powders (single crystal diamond powder, MSY 0-0.03 μm and MSY 0-0.05 μm) can be purchased from Microdiamant, Inc., USA. Anhydrous dichloromethane (≥99.8%, Product #270997), boron tribromide (≥99%, Product #230367), phenylboronic acid pinacol ester (97%, Product #647098), trans-1-propenylboronic acid pinacol ester (97%, Product #735558), triethylborane (1.0M in tetrahydrofuran, Product #179701), triphenylborane (0.25M in tetrahydrofuran, Product #442445), boron trichloride (1.0M in methylene chloride, Product #178934), trimethyl borate (≥99%, Product #92330), triethyl borate (99%, Product #T59307), tributyl borate (≥99%, Product #90795), tri-tert-butyl borate (98%, Product #179701), trimethylborane (1.0M in methylene chloride, Product #178934), trimethylborate (≥99%, Product #92330), triethylborate (99%, Product #T59307), tributylborate (≥99%, Product #90795), tri-tert-butylbo ... #236608), ammonia (0.4M in tetrahydrofuran #718939), 1,1,1,3,3,3-hexafluoro-2-propanol (≥99% #105228), and propargylamine (98% #P50900) can be purchased from Sigma Aldrich (St. Louis, MO). Boron carbide powder (95% 42 μm #815-96) and 400 mesh copper TEM grids with ultrathin lacey carbon film (#01824) are purchased from Ted Pella (Redding, CA). Boron powder (95% #47303) is purchased from Alfa Inorganics (Ward Hill, MA). Approximately 10 cm (4 in) silicon wafers coated with a 10 nm titanium adhesion layer and 100 nm gold are purchased from LGA Thin Films (Santa Clara, CA). Spectra Tech potassium bromide powder packets (#0016031) are purchased from Thermo Fisher Scientific (Waltham, MA).

[0049] Approximately 30 and 50 nm HPHT nanodiamonds (NDs) are aerobically oxidized using a Thermo Scientific (STF55346COMC-1) three-zone tube furnace. ND powder, approximately 500 mg-600 mg, can be placed in a ceramic boat and inserted into a heating chamber. The NDs are oxidized at 525°C for 5 hours in open air conditions until a tan color is observed. Once the oxidation process is complete, the NDs are then placed in glass scintillation vials and stored in a dry oven (approximately 140°C) to ensure a water-free diamond surface. The ND-OH samples are alcohol-rich and ready for further boron chemistry. The ND-OH samples are transferred into a glovebox environment under inert conditions.

[0050] To ensure complete removal of adsorbed water, equipment, including glassware, is placed in a drying oven at 117 °C for 24 hours before any water-sensitive syntheses. Any additional equipment, such as centrifuge tubes (50 mL polypropylene) and micropipette tips, is placed in a vacuum oven set at 40 °C for 24 hours prior to sample preparation. Triangular boron molecules, such as boron tribromide and boron trichloride, can be utilized due to their high reactivity and ability to conjugate with the surface of NDs. As a result of this high reactivity, especially toward oxygen and water, all syntheses are performed in an inert nitrogen atmosphere glovebox.

[0051] In a typical synthesis, 40 mg of ND-OH was placed in a dry 100 mL round-bottom flask and transferred to an inert atmosphere glovebox. 99% boron tribromide and 1.0 M boron trichloride in methylene chloride were reacted with the ND-OH in the round-bottom flask. 20.8 mL of anhydrous DCM and 0.62 mL of BBr3 were added to the 40 mg of ND-OH, capped with a septum, removed from the glovebox, and immediately sonicated. Immediately after the addition of DCM and B-R3, the reaction vessel (centrifuge tube) was removed from the glovebox and sonicated. For sonication, a cup horn sonicator (Fisher Scientific FB505) was used for 2 minutes at 75% of its full power function of 500 W to aid in solubilizing the colloid. Following cup horn sonication, the colloid was further solubilized using a bath sonicator for 10 minutes at 40 °C.

[0052] The solubilized mixture can be centrifuged at 5,000 rpm for 25 minutes until the triangular, end-boronated FNDs form a pellet. The supernatant is then decanted into waste and analyzed via dynamic light scattering (DLS) to ensure minimal FND loss. Three purification cycles are performed by adding approximately 10 mL of DCM to the centrifuge tube containing the FND pellet, and sonicating and centrifuging the sample again to purify the pellet from unreacted B-R3. After the final purification cycle, the end-boronated FND sample is dried inside an inert atmosphere box using a vacuum pump with a solvent trap.

[0053] To synthesize multilamellar liposomes suitable for drug loading, a water / oil / water emulsion is prepared. First, 25 mg of soybean-based asolectin, 1.5 mg of poly(maleic anhydride-alt-1-octadecene) or PMAO, along with 20 μg of Cy5 fluorescent dye and 700 μg of chloroform, are added to a 5 mL centrifuge tube. 200 μg of boron-ND complex is dispersed in 200 μg of water and added to the same Eppendorf tube. The mixture is then probe sonicated at 21% power in an ice bath at 4°C (1 second sonication, followed by a 1 second pause). After sonication, 2 mL of a 0.5 mg / mL aqueous solution of polyethylene glycol (PEG) is quickly added to the mixture, followed by another 1 minute of sonication. The mixture is then stirred for approximately 12 hours until the suspension becomes clear and the chloroform has evaporated from the emulsion. The resulting product is then dialyzed overnight with distilled H2O to produce drug-loaded multilamellar liposomes. To further purify the multilamellar structures, an extruder coupled with a 100-200 nm filter can be used to obtain unilamellar liposomes for encapsulating nanodiamond structures.

[0054] [Table 1-1]

[0055] [Table 1-2]

[0056] Data Analysis and Results DRIFTS data Mechanistically, it was hypothesized that reactive trigonal boron compounds, such as BBr3, attack the hydroxylated surface of the nanodiamond core. This leads to the formation of soluble diatoms, such as Br2 or Cl2, and the desired B-C surface bonds. Residual halides are stripped from the boron to form soluble weak acids, such as ClOH and BrOH. To confirm the presence of B-C bonds, the vibrational modes of the oxidized samples were analyzed using diffuse reflectance Fourier transform infrared spectroscopy (DRIFTS). This was confirmed by the presence of a prominent peak at wavenumber 1105, corresponding to a C-O surface bond. We also observed an OH bending at wavenumber approximately 1640 and a broad OH stretch from wavenumbers 3000-3500. Additionally, a smaller carboxylic acid peak at wavenumber 1800 was observed. After synthesis was completed, we observed a significant shift of the C-O peak at wavenumber 1100 to wavenumber approximately 1025, corresponding to a B-C surface bond. DRIFTS was used to confirm the successful templating of boron onto nanodiamond cores.

[0057] Samples were reacted with various triangular boron precursors, including BBr3, BCl3, and BH3, for various times. We hypothesized that increasing the reaction time from 30 minutes to 180 minutes (Figure 5) and up to 24 hours (Figure 6) would have a measurable effect on the extent of boron templating. While DRIFTS was able to confirm the presence of C-B bonds, further data was needed to quantify the extent of boron templating on the nanodiamond cores.

[0058] SEM, TEM, and EDS data To further confirm the success of the templating, TEM and SEM images of the boron-templated nanodiamond samples were also collected. The TEM data provided some qualitative confirmation that the synthesis process was modifying the diamond surface. Compared to the unreacted oxidized nanodiamond control (Figure 7), there is a significant change in the morphology of the diamonds treated with the boron precursor (Figure 8).

[0059] To further quantify the extent of boron templating, SEM and EDS data were collected. EDS data was used to begin to understand the extent of templating and to quantify the atomic composition of boron at the nanodiamond surface. EDS data was notoriously difficult to collect due to the difficulty in resolving the boron and carbon signals (Figure 10). EDS color mapping indicated that some boron was present on the sample surface (Figure 9). It was hypothesized that varying the reaction time or increasing the concentration of the boron reagent used in the synthesis would affect the extent of templating. More XPS and XAS data were collected to quantitatively assess the extent of templating.

[0060] We initially hypothesized that varying the reagent concentration or increasing the reaction time would result in a larger shell of boron on the ND surface. XPS data and XAS collected to map the elements on our samples showed different results. Spectroscopic analysis seemed to indicate that boron oxide, as opposed to a boron shell coating, was present in our samples. In almost all samples, the XPS and XAS data showed a significant signal for boron oxide, regardless of which reagent was used or how long the samples were reacted (Figure 11). SEM images indicate a significant change in morphology after treatment with the boron precursor, while the XPS and EDS data seem to indicate the presence of a thin boron oxide layer. This contradicts our hypothesis that there was more boron templating in samples reacted for longer times.

[0061] Liposome characterization After confirming the success of boron templating, we began research into liposome fabrication. First, liposomes were synthesized without boronated NDs and unreacted oxidized nanodiamonds. DLS data was collected to determine the particle size distribution in the fabricated liposomes. The collected DLS data showed that the particle size was less than 100 nm, indicating that the emulsion contained some free nanodiamonds, micelles, or empty liposomes. Purification by extrusion dramatically reduced the polydispersity of the sample (Figure 13).

[0062] Liposomes were treated with the fluorescent dye Lucifer Yellow and imaged using a confocal microscope. The confocal images show spherical structures and dark spots, assumed to be free B-ND or unoccupied space (Figure 16).

[0063] Trigonal planar boranes have been shown to modify the surface chemistry of hydroxylated-terminated diamonds. The reactivity of these compounds with ND surfaces can be directly correlated to the size and electronegativity of the -R group on the borane. Further analysis may be required to determine the true composition of the surface structures formed and to gain a better understanding of how organoborane functionalization of FNDs affects the fluorescence on NV centers and how to better control the degree of boron templating. Future research will focus on utilizing this method to manipulate the trigonal planar organoborane modification of stabilizing and bioreactive species. Catalysts can also be used to enhance the degree of boron coating, ensuring that thicker shells can be templated onto the nanodiamond core. Liposome preparation was analyzed by DLS and fluorescence microscopy to confirm that boronated FNDs could be encapsulated into unilamellar liposomes. This suggests that nanoparticles templated with a boron layer encapsulated by liposomes can be used to target tumor sites. 10 This makes them attractive vectors for the delivery of B and ultimately improves their potential to replace older generation BNCT agents (such as boron clusters or substituted amino acids).

[0064] The terms and expressions which have been employed are used as terms of description rather than of limitation, and there is no intention in the use of such terms and expressions to exclude any equivalents of the features or portions thereof shown and described, but it is recognized that various modifications are possible within the scope of the aspects of the invention. Thus, while the present invention has been specifically disclosed by certain embodiments and optional features, it is understood that modifications and variations of the concepts disclosed herein may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of the aspects of the invention.

[0065] A method for forming unique boron bonds on chemically inert diamond surfaces using triangular boron compounds at room temperature 25 nm nanodiamonds were oxidized at high temperatures in open air, resulting in the pyrolysis of the amorphous black carbon and OH surface termination of the diamond, resulting in a highly hydrophilic, tan powder material. Diffuse reflectance Fourier transform infrared spectroscopy (DRIFTS) was used to analyze the surface vibrational modes of the NDOH sample, which was confirmed to have a highly alcohol-rich surface. We observed a peak at 1105 cm, corresponding to a CO surface bond. -1 We also observed a significant peak at 1640 cm -1 OH bending angle at 3000cm -1 -3500cm -1 We observed broad OH stretches from 1785 cm, both of which were likely due to adsorbed water. -1 A smaller carboxylic acid peak at 1640 cm was observed. Temperature-controlled DRIFTS confirmed that both the OH stretch and bending were due to water. We observed 72% desorption of surface water at the experimental temperature of 100 °C, and by the time the sample was raised to 200 °C, more than 97% of the OH bending and stretching corresponding to the adsorbed water had disappeared. Because our starting material (a triangular boron compound) is sensitive to water, and boron trihalides in particular are sensitive to water, we observed a peak at 1640 cm.-1 The peak at 1105 cm is used to roughly quantify the effectiveness of water desorption from our samples. If significant amounts of water are observed in the untreated NDOH sample, we dry the sample overnight at 120°C under vacuum to remove most of the adsorbed water and ensure that the adsorbed water does not lead to degradation of our starting chemicals. Degradation can lead to by-products and, in some cases, the entire boronation / boronation synthesis can fail. After the synthesis is complete, we measure the peak at 1105 cm -1 The CO peak at approximately 1025 cm corresponds to the B-C surface bond. -1 In previous experiments, we observed that samples that still contained adsorbed water formed much fewer BC bonds and instead formed soluble borates such as BOH, which were then washed away in the purification step. If the NDOH sample was not sufficiently dried, it was actually observed that the 1000 cm -1 -1100cm -1 There is a convergence of both the BC and CO peaks in the region, and the drier the sample, the more the BC peak dominates.

[0066] The formation of BC bonds on the surface of NDOH was achieved by the reaction of boron trihalides (BBr and BCl) with NDOH under inert conditions at room temperature, using 0.302 M BX3 and 0.777 mg / mL NDOH. We observed that the synthesis itself was highly sensitive to air and water, as the starting chemicals readily degraded under non-inert conditions. On the other hand, the final product formed demonstrated the stability of the BC bonds when exposed to air and water. Further studies using temperature-controlled DRIFTS may be expected to explore the thermal stability of these bonds and provide a better understanding of the reaction mechanism.

[0067] Exemplary Embodiments The following exemplary aspects are provided, the numbering of which should not be construed as designating a level of importance.

[0068] [Aspect 1] A functionalized nanoscale substrate comprising a functionalized surface, The functionalized surface of the substrate comprises: A functionalized nanoscale substrate comprising a boronated moiety.

[0069] [Aspect 2] The boronated moiety is 10 B. The functionalized nanoscale substrate of embodiment 1, comprising boron.

[0070] [Aspect 3] 3. The functionalized nanoscale substrate of any one of the preceding claims, wherein the substrate comprises diamond, gold, silver, silica, or a mixture thereof.

[0071] [Aspect 4] Aspect 4. The functionalized nanoscale substrate of any one of aspects 1 to 3, wherein the substrate comprises diamond.

[0072] [Aspect 5] 5. The functionalized nanoscale substrate of any one of embodiments 1-4, wherein the boronated moieties are disposed over about 40% to about 100% of the total surface area of ​​the surface of the substrate.

[0073] [Aspect 6] 6. The functionalized nanoscale substrate of any one of embodiments 1-5, wherein the boronated moieties are disposed over about 60% to about 100% of the total surface area of ​​the surface of the substrate.

[0074] [Aspect 7] A functionalized nanoscale substrate comprising: 10 A functionalized nanoscale substrate comprising a boronated surface comprising B boron, said nanoscale substrate comprising diamond.

[0075] [Aspect 8] Aspect 8. The functionalized nanoscale substrate of any one of aspects 1 to 7, wherein a major dimension of the functionalized nanoscale substrate ranges from about 5 nm to about 100 nm.

[0076] [Aspect 9] Aspect 9. The functionalized nanoscale substrate of any one of aspects 1 to 8, wherein a major dimension of the functionalized nanoscale substrate ranges from about 15 nm to about 80 nm.

[0077] [Aspect 10] A delivery vehicle comprising: A delivery vehicle comprising a liposome encapsulating a functionalized nanoscale substrate according to any one of embodiments 1-9.

[0078] [Aspect 11] 11. A method of making a functionalized nanoscale substrate according to any one of embodiments 1 to 10, comprising: A method comprising a boronation step of boronating a hydroxylated nanoscale substrate.

[0079] [Aspect 12] 12. The method of embodiment 11, wherein the boronating step comprises reacting the hydroxylated nanoscale substrate with boron tribromide, boron trichloride, or a mixture thereof.

[0080] [Aspect 13] 11. A method of making a delivery vehicle according to embodiment 10, comprising: contacting the functionalized nanoscale substrate with one or more lipids; mixing the functionalized nanoscale substrate with one or more lipids to form one or more liposomes encapsulating the functionalized nanoscale substrate.

[0081] [Aspect 14] 14. The method of embodiment 13, wherein the one or more lipids comprise one or more phospholipids. [Aspect 15] 1. A method of treating a tumor, comprising: administering a therapeutically effective amount of the functionalized nanoscale substrate to a patient in need thereof; and exposing the dispensed functionalized nanoscale substrate to electromagnetic radiation.

[0082] [Aspect 16] 16. The method of embodiment 15, wherein the functionalized nanoscale substrate is encapsulated in a liposome.

[0083] [Aspect 17] 17. The method of any one of aspects 15 to 16, wherein the tumor comprises a brain tumor. [Aspect 18] 18. The method of any one of aspects 15 to 17, wherein the tumor comprises cutaneous melanoma or glioblastoma.

Claims

1. A functionalized nanoscale substrate comprising a functionalized surface, The functionalized surface of the substrate comprises: A functionalized nanoscale substrate comprising a boronated moiety.

2. The boronated moiety is １０ 10. The functionalized nanoscale substrate of claim 1, comprising B boron.

3. 10. The functionalized nanoscale substrate of claim 1, wherein the substrate comprises diamond, gold, silver, silica, or a mixture thereof.

4. The functionalized nanoscale substrate of claim 1 , wherein the substrate comprises diamond.

5. 10. The functionalized nanoscale substrate of claim 1, wherein the boronated moieties are disposed over about 40% to about 100% of the total surface area of ​​the surface of the substrate.

6. 10. The functionalized nanoscale substrate of claim 1, wherein the boronated moieties are disposed over about 60% to about 100% of the total surface area of ​​the surface of the substrate.

7. A functionalized nanoscale substrate comprising: １０ A functionalized nanoscale substrate comprising a boronated surface comprising B boron, said nanoscale substrate comprising diamond.

8. 8. The functionalized nanoscale substrate of claim 7, wherein a major dimension of the functionalized nanoscale substrate ranges from about 5 nm to about 100 nm.

9. 8. The functionalized nanoscale substrate of claim 7, wherein a major dimension of the functionalized nanoscale substrate ranges from about 15 nm to about 80 nm.

10. A delivery vehicle comprising: A delivery vehicle comprising a liposome encapsulating the functionalized nanoscale substrate of claim 1.

11. 10. A method of making the functionalized nanoscale substrate of claim 1, comprising: A method comprising a boronation step of boronating a hydroxylated nanoscale substrate.

12. 12. The method of claim 11, wherein the boronating step comprises reacting the hydroxylated nanoscale substrate with boron tribromide, boron trichloride, or a mixture thereof.

13. 11. A method of making the delivery vehicle of claim 10, comprising: contacting the functionalized nanoscale substrate with one or more lipids; mixing the functionalized nanoscale substrate with one or more lipids to form one or more liposomes encapsulating the functionalized nanoscale substrate.

14. 14. The method of claim 13, wherein the one or more lipids comprise one or more phospholipids.

15. 1. A method of treating a tumor, comprising: administering a therapeutically effective amount of the functionalized nanoscale substrate to a patient in need thereof; and exposing the dispensed functionalized nanoscale substrate to electromagnetic radiation.

16. 16. The method of claim 15, wherein the functionalized nanoscale substrate is encapsulated in a liposome.

17. 16. The method of claim 15, wherein the tumor comprises a brain tumor.

18. 16. The method of claim 15, wherein the tumor comprises cutaneous melanoma or glioblastoma.

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