Pharmaceutical nanoparticles showing improved mucosal transport

Related Application This application was filed on May 3, 2012, entitled "Nanocrystals, Compositions, and Methods That Aid Particle Transport in Mucus," which is incorporated herein by reference in accordance with Article 119 (e) of the United States Patent Act. Claim priority under US Patent Provisional Application No. 61/642227.

The present invention generally relates to nanocrystals, compositions, and methods that aid in particle transport in mucus.

The mucous layer present at various sites of invasion, including the eyes, nose, lungs, gastrointestinal tract, and female reproductive system, is inherently sticky and effectively traps pathogens, allergens, and necrotic tissue fragments. It helps protect the body from them by removing the mucus more quickly in place of it. In order to effectively deliver therapeutic, diagnostic or contrasting particles through the mucous membrane, the particles must be able to easily penetrate the mucus layer to avoid sticking to the mucus and rapid elimination by the mucus. .. Evidence of some strains suggests that conventional nanoparticles are incapable of crossing the mucosal barrier. However, polymer nanoparticles modified (covalently or non-covalently) with a special surface coating (degradable or non-degradable) are present in water in physiologically considered mucous samples. It has recently been demonstrated that it can spread as rapidly as it does. Such polymer-based mucous permeable particles (MPPs) can be encapsulated with a variety of therapeutic, contrast, or diagnostic agents to allow application in drug delivery, diagnosis, or contrast.

Nevertheless, polymer-based MPPs may have some intrinsic limitations compared to unencapsulated pharmaceutical particles. These constraints, especially when considering application in drug delivery, are 1) essentially lower drug loading, 2) less convenient dosage form (in the case of polymer nanoparticles, in dry powders). (Because it may need to be reconstituted from its storage form), 3) possible increased toxicity, 4) concerns about chemical and physical stability, and 5) at the time of manufacture. It can be mentioned that the complexity of is increased. Therefore, improving compositions and methods comprising mucous permeable particles is beneficial for the delivery of medicinal products.

<p>This description generally relates to nanocrystals, compositions, and methods that aid in particle transport in mucus. In some embodiments, the compositions and methods include mucous permeable particles (MPP) with no or minimal polymeric carrier. The subject matter of this application, in some cases, includes interrelated products, alternative solutions to specific problems, and / or multiple different uses of structures and compositions.</p>

<p>In one set of embodiments, a method of forming coated particles is provided. The method comprises combining the core particles with a solution containing a surface modifier, the core particles comprising a solid pharmaceutical product or a salt thereof, the pharmaceutical product or a salt thereof being about 1 mg / mL or less in a solution at 25 ° C. Having solubility, the drug or salt thereof constitutes at least about 80% by weight of each core particle. The method also comprises coating the core particles with a surface modifier to form the coated particles, the surface modifier comprising a hydrophilic block-hydrophobic block-hydrophilic block arrangement. The hydrophobic block has a molecular weight of at least about 2 kDa, the hydrophilic block constitutes at least about 15% by weight of the triblock copolymer, the hydrophobic block associates with the surface of the core particles, and the hydrophilic block is Present on the surface of the coated particles, making the coated particles hydrophilic, the coated particles have a relative velocity greater than 0.5 in the mucilage.</p><p>In another set of embodiments, a composition comprising a plurality of coated particles is provided. The coated particles comprise core particles containing a solid drug or salt thereof, the drug or salt having a water solubility of about 1 mg / mL or less at 25 ° C anywhere during the pH range. , Pharmaceuticals or salts thereof make up at least about 80% by weight of core particles. The coated particles also comprise a coating comprising a surface modifier surrounding the core particles, the surface modifier comprising a triblock copolymer comprising a hydrophilic block-hydrophobic block-hydrophilic block arrangement, the hydrophobic block. Has a molecular weight of at least about 2 kDa, hydrophilic blocks make up at least about 15% by weight of triblock copolymers, hydrophobic blocks associate with the surface of core particles, and hydrophilic blocks are the surface of coated particles. The surface modifier is at least about 0.001 molecules / nm on the surface of the core particles, which is present in and makes the coated particles hydrophilic.²It exists at the density of. The coated particles have a relative velocity greater than 0.5 in mucus.</p><p>In another set of embodiments, the method of forming coated particles consists of preparing the drug and precipitating the drug by forming a salt in the presence of a surface modifier in an aqueous solution. Including forming core particles, the salt has less water solubility than the non-salt form of the drug, and the water solubility of the salt is about 1 mg at 25 ° C anywhere during the pH range. Below / mL, the surface modifier is present in the aqueous solution at a concentration of at least about 0.01 (weight / volume)%. The method comprises coating core particles with a surface modifier to form coated particles, the surface modifier comprising a triblock copolymer comprising a hydrophilic block-hydrophobic block-hydrophilic block arrangement. Hydrophobic blocks have a molecular weight of at least about 2 kDa, hydrophilic blocks make up at least about 15% by weight of triblock copolymers, hydrophobic blocks associate with the surface of core particles, and hydrophilic blocks are core particles. Makes the particles present and coated on the surface of the surface hydrophilic. The coated particles have a relative velocity greater than 0.5 in mucus.</p><p>In another set of embodiments, treatment methods are provided. The method comprises administering to the patient or subject in need a composition comprising a plurality of coated particles. The coated particles comprise core particles containing a solid drug or a salt thereof, the drug or salt having a water solubility of about 1 mg / mL or less at 25 ° C anywhere during the pH range. The drug or salt thereof constitutes at least about 80% by weight of the core particles. The coated particles also include a coating comprising a surface modifier surrounding the core particles, the surface modifier comprising a triblock copolymer comprising a hydrophilic block-hydrophobic block-hydrophilic block arrangement, the hydrophobic block. Has a molecular weight of at least about 2 kDa, hydrophilic blocks make up at least about 15% by weight of triblock copolymers, hydrophobic blocks associate with the surface of core particles, and hydrophilic blocks are the surface of coated particles. Hydrophilizes the particles present and coated on the surface, and the surface modifier is at least about 0.001 molecules / nm on the surface of the core particles.²It exists at the density of. The coated particles have a relative velocity greater than 0.5 in mucus.</p><p>Other advantages and novel features of the invention, when considered in conjunction with the accompanying drawings, will become apparent from the following detailed description of the various non-limiting embodiments of the invention. In the event that the present specification and the literature incorporated by reference contain inconsistent and / or conflicting disclosures, the present specification shall prevail. If two or more references incorporated by reference contain conflicting and / or conflicting disclosures, the one with the most up-to-date effective date shall prevail.</p><p>Non-limiting embodiments of the present invention will be described with reference to the accompanying figures as examples, but these figures are schematic and are not intended to be drawn to a constant scale. In the figure, each of the same or nearly identical components exemplified is usually represented by a single number. For clarity purposes, not all ingredients are labeled in all figures, and books are not required to illustrate to the person skilled in the art so that they can understand the invention. Not all components of each embodiment of the invention are shown. The figure is as follows.</p>

<figref num="1">FIG. 3 is a schematic representation of mucus-penetrating particles with a coating and a solid pharmaceutical core according to a set of embodiments.</figref><figref num="2A">200 nm carboxylated polystyrene particles (negative control), 200 nm pegged polystyrene particles (positive control), and nanocrystal particles made by nanogrinding and coated with different stabilizers / surface modifiers according to a set of embodiments. Average velocity of aggregates in human cervical mucus (CVM) with respect to (sample) &lt;V_{mean mean}&gt; Is a plot showing.</figref><figref num="2B">Relative velocity &lt;V in CVM for nanocrystal particles made by nanomilling and coated with different stabilizers / surface modifiers according to a set of embodiments._{mean mean}&gt;_relIt is a plot showing.</figref><figref num="3A">Trajectory average velocity V in CVM within aggregates of nanocrystal particles coated with different surface modifiers according to a set of embodiments_{mean mean}It is a histogram showing the distribution of.</figref><figref num="3B">Trajectory average velocity V in CVM within aggregates of nanocrystal particles coated with different surface modifiers according to a set of embodiments_{mean mean}It is a histogram showing the distribution of.</figref><figref num="3C">Trajectory average velocity V in CVM within aggregates of nanocrystal particles coated with different surface modifiers according to a set of embodiments_{mean mean}It is a histogram showing the distribution of.</figref><figref num="3D">Trajectory average velocity V in CVM within aggregates of nanocrystal particles coated with different surface modifiers according to a set of embodiments_{mean mean}It is a histogram showing the distribution of.</figref><figref num="4">&lt;V in CVM coated with different PEO-PPO-PEO Pluronic® triblock copolymers according to a set of embodiments._{mean mean}&gt;_relIt is a plot showing PPO block and PEO weight content (%) in terms of molecular weight.</figref><figref num="5">According to a set of embodiments, the mass of transport via CVM for solid particles with different cores coated with either Pluronic® F127 (MPP) or sodium dodecyl sulfate (CP, negative control). It is a plot shown.</figref><figref num="6A">New Zealand white rabbit eyelid conjunctiva after administration of Loteprednol etabonate prescription drug, Lotemax®, or according to a set of embodiments, after administration of particles of Loteprednol etabonate coated with Pluronic® F127. FIG. 6A) shows the drug level of rotepredonor etabonate in.</figref><figref num="6B">New Zealand white rabbit eyeball conjunctiva after administration of Loteprednol etabonate prescription drug, Lotemax®, or according to a set of embodiments, after administration of particles of Loteprednol etabonate coated with Pluronic® F127. FIG. 6B) shows the drug level of rotepredonor etabonate in.</figref><figref num="6C">Loteprednol New Zealand white rabbit corneum after administration of Loteprednol, a prescription drug of Loteprednol etabonate, or after administration of particles of Loteprednol etabonate coated with Pluronic® F127 according to a set of embodiments (Figure). It shows the drug level of rotepredonor etabonate in 6C).</figref><figref num="7A">It is a diagram showing the physicochemical characteristics of CUR-1% F127 particles according to a set of embodiments, and a powder X-ray diffraction (powder-XRD) diagram of F127, raw curcumin, and CUR-1% F127 particles. It is a figure which shows.</figref><figref num="7B">It is a figure which shows the physicochemical characteristics of a CUR-1% F127 particle according to a set of embodiments, and is the figure which shows the typical transmission electron microscope (TEM) image of a CUR-1% F127 particle.</figref><figref num="8A">According to a set of embodiments, the ensemble average square geometric mean displacement of CUR-1% F127 particles, 200 nm carboxylated polystyrene (PSCOOH) particles, and 200 nm PEGylated polystyrene (PSPEG) particles in a CVM as a function of the time scale. It is a figure which shows.</figref><figref num="8B">As a function of the time scale of CUR-1% F127 particles, 200 nm carboxylated polystyrene (PSCOOH) particles, and 200 nm PEGylated polystyrene (PSPEG) particles in human cystic fibrosis sputum (CFS) according to a set of embodiments. It is a figure which shows the ensemble average square geometric mean displacement of.</figref><figref num="9">Geometric ensemble effective diffusion coefficient (&lt;D) on a time scale of 1 second in human CVM for CUR particles formulated in different concentrations of F127 according to a set of embodiments._eff&gt;) Is a plot showing. The data represent an ensemble average of at least three independent experiments with n 100 for each experiment. Error bars indicate geometric standard errors.</figref><figref num="10A">A plot showing the diffusivity of CUR particles formulated with different Pluronic® in human CVM according to a set of embodiments, Pluronic® Poly (ethylene glycol) (PEG). &lt;D on a 1 second time scale for molecular weight (MW) of segments and poly (propylene oxide) (PPO) segments_effIt is a figure which shows the distribution of&gt;. Each data point represents a particular type of Pluronic®. The MW of PPO and PEG was estimated based on the molecular weights presented by the manufacturer.</figref><figref num="10B">A plot showing the diffusivity of CUR particles formulated with different Pluronic® in human CVM according to a set of embodiments, on a time scale of 1 second as a function of MW of PEG segments. &lt;D_effIt is a figure which shows&gt;. The inset shows the same plot on a linear scale &lt;D._eff&gt;, And R represents the correlation coefficient. The data represent an ensemble average of at least three independent experiments with n 100 for each experiment. Error bars indicate geometric standard errors.</figref><figref num="10C">A plot showing the diffusivity of CUR particles formulated with different Pluronic® in human CVM according to a set of embodiments, on a time scale of 1 second as a function of MW in the PPO segment. &lt;D_effIt is a figure which shows&gt;. The inset shows the same plot on a linear scale &lt;D._eff&gt;, And R represents the correlation coefficient. The data represent an ensemble average of at least three independent experiments with n 100 for each experiment. Error bars indicate geometric standard errors.</figref><figref num="11">It is a plot showing the cumulative release of CUR-1% F127 particles in phosphate buffered saline (pH = 7.4) by octanol extraction according to a set of embodiments.</figref><figref num="12">According to a set of embodiments, it is a plot showing the cumulative release of free TFV in solution from a dialysis bag into normal phosphate buffered saline as opposed to that of TFV-Zn particles in suspension.</figref><figref num="13A">It is an image showing the distribution of mucus permeability / F127-coated TFV particles on flattened vaginal tissue from human-like estrous mice according to a set of embodiments. Vaginal tissue was removed within 10 minutes of administration.</figref><figref num="13B">It is an image showing the distribution of mucous adhesive / PVA-coated TFV particles on flattened vaginal tissue from human-like estrous mice according to a set of embodiments. Vaginal tissue was removed within 10 minutes of administration.</figref><figref num="14A">The figure which shows the transport rate of CP and MPP in CVM of the estrus mouse, and the figure which shows the typical locus for the particle which shows the effective diffusion coefficient in one SEM of the ensemble average on the time scale of 1 second. Is.</figref><figref num="14B">It is a figure which shows the transport rate of CP and MPP in CVM of an estrus mouse, and is the figure which shows the ensemble average square geometric mean displacement (&lt;MSD&gt;) as a function of the time scale. Data for particles on in vitro mouse vaginal tissue (mCVM) are shown in the same particles (SKLai, YYWang, K.Hida, R.Cone, J.Hanes, Nanoparticles reveal that human cervicovaginal mucus is riddled with pores larger than viruses.P Natl Acad Sci USA 107,598-603 (2010)), and the theoretical diffusion rate of 110 nm particles in water (about 4 μm)²/ Second) compared.</figref><figref num="14C">It is a figure which shows the transport rate of CP and MPP in the CVM of an estrous mouse, and the effective diffusion coefficient (D) of an individual particle on a time scale of 1 second._effIt is a figure which shows the distribution of the logarithm of). The data represent the ensemble average of three independent experiments with n 150 for each experiment. The value of the diffusivity corresponding to the left of the dotted line indicates a particle with an MSD value less than the particle diameter.</figref><figref num="14D">It is a diagram showing the transport rate of CP and MPP in CVM of estrus mice, and the proportion of particles capable of penetrating into a 100 μm thick layer of mouse CVM over time is measured experimentally. It is a figure shown based on Fick's second law about the diffusion simulation of the particle undergoing random diffusion which has a diffusion coefficient equal to a coefficient.</figref><figref num="14E">It is the figure which illustrated the delivery of the vaginal drug from the gel preparation, the CP preparation, and the MPP preparation.</figref><figref num="15A">Transport of nanoparticles on the vaginal tissue of IE mice, ie plots showing ensemble mean square geometric mean displacement (&lt;MSD&gt;) as a function of time scale. Data are induced estrus (IE) and natural periodic estrus. It is shown for MPP and CP on in vitro vaginal tissue of stage mice.</figref><figref num="15B">Transport of nanoparticles on the vaginal tissue of IE mice, a plot showing the ensemble mean square geometric mean displacement (&lt;MSD&gt;) as a function of the time scale. Biodegradable MPPs on IE tissues with non-biodegradable MPPs. The data represent an ensemble average of at least three independent experiments using an average of n 150 particles and at least 130 particles for each experiment. The data represent the mean standard error as an average. Provided with (SEM).</figref><figref num="16">It is a figure which contains the image which shows the distribution of the particle in the vagina of a mouse. The distribution of red fluorescent non-biodegradable and biodegradable CP and MPP in estrus and in cross-frozen sections of vaginal tissue of IE mice is shown. The image is representative of a mouse with n 3.</figref><figref num="17">FIG. 6 includes images and plots showing quantification of vaginal coverage with nanoparticles. The distribution of red fluorescent non-biodegradable and biodegradable CP and MPP on the flattened vaginal tissue of estrus mice is shown. The inset is an image of a dark area at higher magnification. The image corresponds to the calculated average for mice with n 3. The data are mean ± SEM. * P &lt;0.05, Student's t-test.</figref><figref num="18">It is a figure which includes the image which shows the cervical coverage with nanoparticles. The distribution of red fluorescent non-biodegradable and biodegradable CP and MPP in the cervix on the cervical tissue of estrus mice is shown. The inset is an image of a dark area at higher magnification. The image is representative of a mouse with n 3. The data are mean ± SEM. * P &lt;0.05, Student's t-test.</figref><figref num="19">It is a figure which contains the image which shows the effect of the mucus removal on the mucus sticky nanoparticles. Red fluorescent non-biodegradable and biodegradable CP in cross-frozen sections of vaginal tissue of mice with an intact mucus layer (No treatment) or mucus removed by washing and swab treatment. Shows the distribution. The image is representative of a mouse with n 3.</figref><figref num="20">It is a figure which contains the image which shows the distribution of the particle in the vagina of IE mouse. The distribution of red fluorescent non-biodegradable CP and MPP in cross-frozen sections of vaginal tissue of IE mice is shown. The image is representative of a mouse with n 3.</figref><figref num="21A">FIG. 5 shows images and plots showing retention of non-biodegradable MPP and CP in the cervical vaginal canal of IE mice. The fluorescence intensity of particles related to CP and MPP in all cervical and vaginal tissues and the superimposition of Akeno images are shown.</figref><figref num="21B">FIG. 5 shows images and plots showing retention of non-biodegradable MPP and CP in the cervical vaginal canal of IE mice. It is a figure which shows the fraction of the particle which remains even if the time elapses based on the quantification of the fluorescence of the particle. The data are mean ± SEM (n 7). * P &lt;0.05, Student's t-test.</figref><figref num="22">FIG. 6 includes images showing the distribution and retention of FITC, a model drug delivered in gel form or encapsulated in biodegradable MPP in the vagina of estrus mice. Fluorescent images were taken 24 hours later on the flattened vaginal tissue of the mice. The image is representative of a mouse with n 3. The data are mean ± SEM. * P &lt;0.05, Student's t-test.</figref><figref num="23">It is a figure which contains the image which shows the acute toxicity and the cytokine concentration by the daily administration. FIG. 6 shows a hematoxylin and eosin (H &amp; E) -stained cross section of vaginal tissue of DP mice removed 24 hours after intravaginal administration of 5% N9, PBS, CP, MPP, BD-CP, and BD-MPP. The scale bar applies to all images. Arrows indicate clusters of neutrophils. The image is representative of a mouse with n 5.</figref><figref num="24">It is a plot which shows the cytokine concentration by the daily administration. Cytokine levels in cervical lavage (CVL) of DP mice after daily vaginal treatment for 7 days. The data are mean ± SEM. * P &lt;0.05, Student's t-test.</figref><figref num="25">FIG. 6 is a bar graph showing the relationship between the relative velocities of polystyrene particles coated with Pluronic® F127 in mucus and the density of Pluronic® F127 molecules on the particle surface.</figref>

Nanocrystals, compositions and methods that aid in particle transport in mucus are provided. In some embodiments, the composition and method comprises making mucous permeable particles (MPP) with no polymeric carrier or with minimal polymeric carrier. The compositions and methods can include, in some embodiments, modifying the surface coating of particles formed from a pharmaceutical product having low solubility in water / aqueous solutions. Such methods and compositions are used to achieve efficient transport of pharmaceutical particles in a mucous barrier throughout the body for a wide range of applications, including drug delivery, contrast and diagnostic applications. be able to. In certain embodiments, pharmaceutical compositions containing such particles are suitable for routes of administration that require particles that cross the mucosal barrier.

In some embodiments, the particles described herein have a core-shell configuration. The wick can contain a solid drug or a salt thereof, which has a relatively low water solubility. The wick can also contain a gel or liquid in some embodiments. As will be described in more detail later, in some embodiments, the surface modifier can include a triblock copolymer with a hydrophilic block-hydrophobic block-hydrophilic block arrangement. The molecular weight of each hydrophilic and hydrophobic block can be selected to confer specific transport properties, eg, increased transport in mucus, to the particles.

This provides a non-limiting example of particles. As shown in the exemplary embodiment of FIG. 1, the particles (10) include a core (16) (which may be in the form of particles, referred to herein as core particles) and a coating (20) surrounding the core. .. In a set of embodiments, a substantial portion of the core is one or more solid medicines (eg, drugs, therapeutic agents, diagnostic agents, contrasts) that can provide a particular beneficial and / or therapeutic effect. Drugs). The core may be, for example, pharmaceutical nanocrystals (ie, nanocrystalline particles). The wick comprises a surface (24) to which one or more surface modifiers can be attached. For example, in some cases, the core (16) is surrounded by a coating (20) with an inner surface (28) and an outer surface (32). The coating can be formed, at least in part, from one or more surface modifiers (34), such as polymers (eg, block copolymers) that can be attached to the surface of the core (24). The surface modifier (34) can be, for example, covalently bound to the core particles, non-covalently bound to the core particles, adsorbed to the core, or ion interaction, hydrophobic and / or. It can associate with core particles by being attached to the core via hydrophilic interactions, electrostatic interactions, van der Waals interactions, or a combination thereof. In a set of embodiments, the surface modifier or a portion thereof is selected to facilitate the transport of particles in a mucus barrier (eg, mucus or mucosa).

In certain embodiments described herein, one or more surface modifiers (34) are oriented in a particular arrangement during coating of particles. For example, in some embodiments where the surface modifier is a triblock copolymer such as a triblock copolymer having a hydrophilic block-hydrophobic block-hydrophilic block arrangement, the hydrophobic block 36 is directed towards the surface of the core. The hydrophilic block 38 can be oriented away from the surface of the core (eg, towards the outside of the particles). Hydrophilic blocks can have properties that facilitate the transport of particles in the mucosal barrier, as will be described in more detail later.

The particle (10) can optionally contain one or more components (40) such as targeted moieties, proteins, nucleic acids, and bioactive agents that can optionally impart specificity to the particle. .. For example, a targeting agent or molecule (eg, a protein, nucleic acid, nucleic acid analog, carbohydrate, or small molecule), if present, can help guide the particle to a specific location in the subject's body. .. The location can be, for example, a tissue, a particular cell type, or an intracellular compartment. One or more components (40), if present, can associate with the core, coating, or both, for example, those components are the surface of the core (24), the inner surface of the coating (28). , Can associate with the outer surface (32) of the coating and / or be embedded in the coating. One or more components (40) may be associated through covalent bonds, absorption, or ion interactions, hydrophobic and / or hydrophilic interactions, electrostatic interactions, van der Waals interactions, or them. Can be attached via a combination of. In some embodiments, the ingredients can be attached (eg, by covalent bond) to one or more surface modifiers of the coated particles using methods known to those of skill in the art.

Ingredients and arrangements other than those shown in FIG. 1 or described herein may be suitable for a particular particle and composition, and all of the ingredients shown in FIG. 1 are in part. It should be understood that it does not always exist in the embodiments.

In one set of embodiments, the particles (10), when introduced into a subject, are one or more components in the subject, such as mucus, cells, tissues, organs, particles, body fluids (eg, blood). , These parts, and combinations thereof. In some such embodiments, the coating of the particle (10) has a surface modification that has the property of allowing favorable interaction (eg, transport, binding, adsorption) with one or more materials from the subject. It can be designed to contain agents or other ingredients. For example, the coating may be a surface modifier or other having specific hydrophilicity, hydrophobicity, surface charge, functional groups, binding specificity, and / or density to promote or reduce specific interactions within the subject. Can contain the components of. One particular example is one or more surface modifiers to reduce the physical and / or chemical interaction between the particles and the mucous of interest in order to enhance the movement of the particles in the mucilage. Includes selecting specific hydrophilicity, hydrophobicity, surface charge, functional groups, binding specificity, and / or density of. Other examples will be described in more detail later.

In some embodiments, when the particles are successfully transported across a mucosal barrier in the subject (eg, mucus or mucosa), further interactions between the particles in the subject can occur. Interactions can occur through the coating and / or core in some cases, eg, from one or more components of interest to and / or from the particle (10). It can include the exchange of materials (eg, pharmaceuticals, therapeutic agents, proteins, peptides, polypeptides, nucleic acids, nutrients, etc.) to one or more components of interest. For example, in some embodiments where the wick is formed from a drug or contains a drug, disintegration, release and / or transport of the drug from the particles leads to certain beneficial and / or therapeutic effects in the subject. Sometimes. Thus, the particles described herein can be used for the diagnosis, prevention, treatment or management of a particular disease or physical condition.

Specific examples of the use of the particles described herein are provided later in the context of being suitable for administration to a mucosal barrier (eg, mucus or mucosa) in a subject. Although many embodiments herein are described in this context and in the context of providing benefits for diseases and conditions that require transport of the material across the mucosal barrier, the invention is described. Recognize that the particles, compositions, kits, and methods described herein can be used to prevent, treat, or control other diseases or conditions. I want to be.

Mucus is a sticky viscoelastic gel that protects against pathogens, toxins, and necrotic tissue debris at various points of entry into the body, including the eyes, nose, lungs, gastrointestinal tract, and female reproductive system. Many synthetic nanoparticles are strongly mucous adhesive to mucus and are effectively trapped in the rapidly eliminated peripheral mucous membranes, distributing them throughout the mucosa and penetrating towards the underlying tissue. Is greatly constrained. The residence time of these captured particles is constrained by the rate of replacement of the peripheral mucus layer ranging from seconds to hours, depending on the organ. To ensure effective delivery of particles containing pharmaceuticals (eg, therapeutic, diagnostic and / or contrasting agents) through the mucus membrane, such particles easily diffuse in the mucus barrier and adhere to the mucus. Must be able to be avoided.

By modifying the surface of the polymeric nanoparticles with a mucous permeable coating, sticking to the mucus can be minimized, thereby allowing rapid particle penetration across the mucous barrier. Specifically, polymer nanoparticles of about 500 nm are covalently bonded to a dense coating of low molecular weight PEG (2 kDa to 5 kDa) or non-covalently bonded specific Pluronic® molecules (eg, P103, P105). , F127), at about the same speed as those nanoparticles move in pure water, and almost 100 times faster than uncoated polymer particles of similar size. It has been shown to be able to penetrate human mucus.

However, polymer-based mucous permeable particles may have one or more inherent constraints in some embodiments. Among other things, considering their application to drug delivery, these constraints may include one or more of the following: A) Low drug encapsulation efficiency and low drug loading: Encapsulation in particles during production. Encapsulation of the drug in the polymeric particles is often inefficient, as it is generally less than 10% of the total amount of drug used. Moreover, drug loading greater than 50% is rarely achieved. B) Ease of use: In general, pharmaceutical-laden polymer particle-based formulations typically require storage as a dry powder to avoid premature drug release, and therefore at the time of use. Requires either reconstitution in or complex dosing devices. C) Biocompatibility: Accumulation of polymeric carriers that gradually degrade following repeated doses and their toxicity over time presents significant concerns for polymeric drug carriers. D) Chemical and physical stability: Degradation of the polymer can jeopardize the stability of the encapsulated drug. In many encapsulation steps, the drug experiences a transition from the solution phase to the solid phase, which transition is sufficient for the physical morphology of the emerging solid phase (eg, amorphous / crystalline / crystalline polymorph). Not controlled by. This is a concern for physical and chemical stability, as well as the performance of various aspects of the product, including release kinetics. E) Manufacturing Complexity: The production of drug-loaded polymeric MPPs, especially large-scale feasibility, is usually a fairly complex process involving a large number of steps and a significant amount of toxic organic solvent.

In some embodiments described herein, the method for preparing a composition and particles, eg, a particular composition, and a method for preparing particles with enhanced transport within a mucosal barrier, are described above. Address one, more, or all of your concerns. Specifically, in some embodiments, the compositions and methods do not require, if at all, encapsulation in a polymeric carrier. Advantageously, drug loading, convenience of use, biocompatibility, stability, by avoiding or minimizing the need for encapsulation of the drug (eg, drug, contrast or diagnostic agent) in a polymeric carrier. And / or the specific limitations of polymeric MPPs with respect to manufacturing complexity can be addressed. The methods and compositions described herein have the potential to facilitate the clinical development of mucus permeable particle technology.

Core Particles As previously described with respect to FIG. 1, particles (10) can include cores (16). The wick can be formed from any suitable material such as an organic material, an inorganic material, a polymer, or a combination thereof. In one set of embodiments, the wick comprises a solid. The solid may be, for example, a crystalline or amorphous solid, such as a crystalline or amorphous solid drug (eg, a therapeutic agent, a diagnostic agent, and / or a contrast agent) or a salt thereof. In some embodiments, there can be more than one drug in the core. Specific examples of pharmaceutical products will be presented in more detail later.

The drug is in any suitable amount in the wick, eg, at least about 1% by weight, at least about 5% by weight, at least about 10% by weight, at least about 20% by weight, at least about 30% by weight, at least about 40%. %%, at least about 50% by weight, at least about 60% by weight, at least about 70% by weight, at least about 80% by weight, at least about 85% by weight, at least about 90% by weight, at least about 95% by weight, or at least about 99% by weight Can exist in quantity of%. In one embodiment, the wick is formed from 100% by weight medicinal products. In some cases, the drug is in the core about 100% by weight or less, about 90% by weight or less, about 80% by weight or less, about 70% by weight or less, about 60% by weight or less, about 50% by weight or less, about It can be present in 40% by weight or less, about 30% by weight or less, about 20% by weight or less, about 10% by weight or less, about 5% by weight or less, about 2% by weight or less, or about 1% by weight or less. Combinations in the range mentioned above are also possible (eg, present in an amount of at least about 80% by weight and less than about 100% by weight). Other ranges are possible.

In embodiments that include a drug with a relatively large amount of core particles (eg, at least about 50% by weight of the core particles), the core particles are compared to the particles formed by encapsulating the drug in a polymeric carrier. Generally has an increased drug load. This means that a higher drug load means that fewer particles are required to achieve the desired effect compared to the use of particles containing a polymeric carrier. Convenient for application to drug delivery.

In some embodiments, the wick is relatively low in water solubility (ie, soluble in water containing one or more buffers in some cases), and / or the solid material is coated with a surface modifier. It can be formed from a solid material that has relatively low solubility in the solution. For example, solid materials at 25 ° C, about 5 mg / mL or less, about 2 mg / mL or less, about 1 mg / mL or less, about 0.5 mg / mL or less, about 0.1 mg / mL or less, about 0.05 mg / mL or less, Water solubility of about 0.01 mg / mL or less, about 1 μg / mL or less, about 0.1 μg / mL or less, about 0.01 μg / mL or less, about 1 ng / mL or less, about 0.1 ng / mL or less, or about 0.01 ng / mL or less Can have (or solubility in a coating solution). In some embodiments, the solid material is at least about 1 pg / mL, at least about 10 pg / mL, at least about 0.1 ng / mL, at least about 1 ng / mL, at least about 10 ng / mL, at least about 0.1 μg / mL, at least about 0.1 μg / mL. About 1 μg / mL, at least about 5 μg / mL, at least about 0.01 mg / mL, at least about 0.05 mg / mL, at least about 0.1 mg / mL, at least about 0.5 mg / mL, at least about 1.0 mg / mL, at least about 2 mg / mL It can have mL water solubility (or solubility in a coating solution). Combinations in the range mentioned above (eg, solubility in water-soluble or coating solutions of at least about 10 pg / mL and about 1 mg / mL or less) are also possible. Other ranges are possible. Solid materials can have water solubility in these or other ranges at any point during the pH range (pH 1 to pH 14).

In some embodiments, the wick is classified by US Pharmacopeia practice: range of solubility: highly soluble:&gt; 1,000 mg / mL, soluble: 100-1,000 mg / mL, slightly soluble: 33 ~ 100mg / mL, slightly insoluble: 10 ~ 33mg / mL, difficult to dissolve: 1 ~ 10mg / mL, extremely difficult to dissolve: 0.1 ~ 1mg / mL, and almost insoluble: included in one of &lt;0.1mg / mL It can be formed from the material to be made.

The wick may be hydrophobic or hydrophilic, but in many embodiments described herein, the wick is substantially hydrophobic. "Hydrophobic" and "hydrophilic" are given the usual meanings in the art as understood by those of skill in the art and are relative terms in many of the examples herein. The relative hydrophobicity and hydrophilicity of the material is measured by measuring the contact angle of the water droplet on a plane consisting of the substance to be measured using a device such as a contact angle measuring instrument and the powder of the compacted core material. It can be obtained by.

In some embodiments, the material (eg, the material that forms the core of the particle) is at least about 20 °, at least about 30 °, at least about 40 °, at least about 50 °, at least about 60 °, at least about 70 °. Has a contact angle of at least about 80 °, at least about 90 °, at least about 100 °, at least about 110 °, at least about 120 °, or at least about 130 °. In some embodiments, the material is about 160 ° or less, about 150 ° or less, about 140 ° or less, about 130 ° or less, about 120 ° or less, about 110 ° or less, about 100 ° or less, about 90 ° or less, It has a contact angle of about 80 ° or less, or about 70 ° or less. Combinations in the range mentioned above (eg, contact angles of at least about 30 ° and about 120 ° or less) are also possible. Other ranges are possible.

The measurement of the contact angle can be performed using various techniques, and here we refer to the measurement of the static contact angle between the pellet of starting material and the water ball used to form the core. .. The material used to form the wick was either accepted as a fine powder or otherwise ground into a fine powder using a mortar and pestle. International Crystal to form a surface on which to make measurements The powder was compacted using a 7 mm pellet mold set from Labs. The material was packed in a mold and the powder was compacted into pellets by applying pressure by hand without using a pellet press or high pressure. The pellet was then floated for testing so that the top and bottom surfaces of the pellet (defined as the surface to which water is applied and the opposite surface parallel to each other) do not touch any surface. This was done by not removing the pellet completely from the collar of the mold set. Therefore, the pellet contacts the collar on the sides and not on the top and bottom. When measuring the contact angle, water was added to the surface of the pellet for at least 30 seconds until a water ball with a stable contact angle was obtained. Water was added into the water ball by inserting or contacting the tip of a pipette or syringe used to add to the water ball into the water ball. Once a stable water ball was obtained, an image was taken and the contact angle was measured using standard means.

In embodiments where the core comprises an inorganic material (eg, for use as a contrasting agent), the inorganic material may include, for example, a metal (eg, Ag, Au, Pt, Fe, Cr, Co, Ni, Cu, Zn. , And other transition metals), semiconductors (eg, silicon, silicon compounds and alloys, cadmium selenium, cadmium sulfide, indium arsenide, and indium phosphate), or insulating materials (eg, ceramics such as silicon oxide). be able to. The inorganic material is in the core in any suitable amount, eg, at least about 1% by weight, at least about 5% by weight, at least about 10% by weight, at least about 20% by weight, at least about 30% by weight, at least about 40%. It can be present in an amount of% by weight, at least about 50% by weight, at least about 75% by weight, at least about 90% by weight, or at least about 99% by weight. In one embodiment, the wick is formed from 100% by weight inorganic material. In some cases, the inorganic material in the core is about 100% by weight or less, about 90% by weight or less, about 80% by weight or less, about 70% by weight or less, about 60% by weight or less, about 50% by weight or less, It can be present in an amount of about 40% by weight or less, about 30% by weight or less, about 20% by weight or less, about 10% by weight or less, about 5% by weight or less, about 2% by weight or less, or about 1% by weight or less. .. Combinations in the range mentioned above (eg, present at least about 1% by weight and less than about 20% by weight) are also possible. Other ranges are possible.

The wick can, in some cases, be in the form of quantum dots, carbon nanotubes, carbon nanowires, or carbon nanorods. In some cases, the wick contains or is formed from a material that is not of biological origin.

In some embodiments, the wick comprises one or more organic materials such as synthetic and / or natural polymers. Examples of synthetic polymers include non-degradable polymers such as polymethacrylates and degradable polymers such as polylactic acid, polyglycolic acid and copolymers thereof. Examples of natural polymers include hyaluronic acid, chitosan, and collagen. Other examples of polymers that may be suitable for the core portion include the following, which are suitable for forming coatings on particles. The polymer is in any suitable amount in the core, eg, about 100% by weight or less, about 90% by weight or less, about 80% by weight or less, about 70% by weight or less, about 60% by weight or less, about 50% by weight. Below, it must be present in an amount of about 40% by weight or less, about 30% by weight or less, about 20% by weight or less, about 10% by weight or less, about 5% by weight or less, about 2% by weight or less, or about 1% by weight or less. Can be done. In some cases, the polymer is at least about 1% by weight, at least about 5% by weight, at least about 10% by weight, at least about 20% by weight, at least about 30% by weight, at least about 40% by weight, at least about 50%. It can be present in an amount of% by weight, at least about 75% by weight, at least about 90% by weight, or at least about 99% by weight. Combinations in the range mentioned above are also possible (eg, present in an amount of at least about 1% by weight and about 20% by weight or less). Other ranges are possible. In a set of embodiments, the wick is formed from or virtually free of polymer components.

The wick can have any suitable shape and / or size. For example, the core may be substantially spherical, non-spherical, oval, rod-shaped, pyramidal, cubic, disc-shaped, wire-shaped, or irregularly shaped. The core is, for example, about 10 μm or less, about 5 μm or less, about 1 μm or less, about 800 nm or less, about 700 nm or less, about 500 nm or less, 400 nm or less, 300 nm or less, about 200 nm or less, about 100 nm or less, about 75 nm or less, about 50 nm or less. Can have maximum or minimum cross-sectional dimensions of about 40 nm or less, about 35 nm or less, about 30 nm or less, about 25 nm or less, about 20 nm or less, about 15 nm or less, or about 5 nm or less. In some cases, the core is, for example, at least about 5 nm, at least about 20 nm, at least about 50 nm, at least about 100 nm, at least about 200 nm, at least about 300 nm, at least about 400 nm, at least about 500 nm, at least about 1 μm, or at least about about. It can have a maximum or minimum cross-sectional dimension of 5 μm. Combinations in the range mentioned above (eg, maximum or minimum cross-sectional dimensions of at least about 50 nm and about 500 nm or less) are also possible. Other ranges are possible. In some embodiments, the size of the core formed by the method described herein has a Gaussian distribution. Unless otherwise stated, particle / core size measurements herein refer to minimum cross-sectional dimensions.

Those of skill in the art are familiar with techniques for measuring particle size (eg, minimum or maximum cross-sectional dimensions). Examples of suitable techniques include (DLS), transmission electron microscopy, scanning electron microscopy, electrical resistance counting, and laser diffraction, and other suitable techniques are also known to those of skill in the art. Although many methods for measuring particle size are known, the sizes described herein (eg, average particle size, thickness) are those measured by dynamic light scattering. be.

Methods for Forming Core Particles and Coated Particles The core particles described herein can be formed by any suitable method. In some embodiments, a grinding step is used to reduce the size of the solid material to form particles with a size range of micrometer to nanometer. Dry and wet grind methods such as jet grind, freeze grind, ball grind, medium grind, and homogenization are known and can be used in the methods described herein. Generally, in the wet grinding method, a suspension of the material to be used as a core is mixed with or without an excipient with a grinding medium to reduce the particle size. Dry grinding is a method of mixing a material to be used as a core with a grinding medium with or without an excipient to reduce the particle size. In the freeze grind method, a suspension of the material to be used as the core is mixed with the grind medium with or without the excipient at a cooled temperature.

In some embodiments, the core particles described herein can be made by nanogrinding a solid material (eg, a pharmaceutical product) in the presence of one or more stabilizers / surface modifiers. .. Small particles of solid material allow the presence of one or more stabilizers / surface modifiers, especially on the surface of the particles, to stabilize the suspension of the particles without aggregation or agglomeration in a liquid solution. May be needed. In some such embodiments, the stabilizer can act as a surface modifier to form a coating on the particles.

As described herein, in some embodiments, the method of forming core particles is to select a stabilizer suitable for both nanogrinding and forming a coating on the particles, and the particles. Includes making mucus permeable. For example, as will be described in more detail later, the 200-500 nm nanoparticles of the model compound pyrene produced by nanogrinding pyrene in the presence of Pluronic® F127 are polymer-based and well established. It has been demonstrated to provide particles that can penetrate physiological mucus samples at a rate similar to that of MPPs. Interestingly, as will be described in more detail later, only a small amount of stabilizer / surface modifier tested is nanomilled and forms a coating on the particles that makes them mucous permeable. It was observed that the criteria were met as being suitable for both.

In the wet grinding method, grinding is a dispersion containing one or more stabilizers (eg, surface modifiers), a grinding medium, a solid to be ground (eg, solid pharmaceuticals), and a solvent (eg, aqueous dispersion). Can be carried out in liquid). Any suitable amount of stabilizer / surface modifier can be included in the solvent. In some embodiments, the stabilizer / surface modifier is at least about 0.001%, at least about 0.01%, at least about 0.1% of the solvent in% by weight or% by weight (weight: volume). At least about 0.5%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 10%, It can be present in an amount of at least about 12%, at least about 15%, at least about 20%, at least about 40%, at least about 60%, or at least about 80%. In some cases, the stabilizer can be present in the solvent in an amount of about 100% (eg, in the case where the stabilizer / surface modifier is the solvent). In other embodiments, the stabilizer is dissolved.In the medium, about 100% or less, about 80% or less, about 60% or less, about 40% or less, about 20% or less, about 15% or less, about 12% or less, about 10% or less, about 8% or less. Can be present in about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less. Combinations in the range mentioned above (eg, about 5% or less of the solvent and at least about 1%) are also possible. Other ranges are possible. The specific range selected is a factor that can affect the ability of the particles to penetrate the mucus, such as the stability of the coating consisting of stabilizers / surface modifiers on the particle surface, stabilizers on the particles / Average thickness of coating consisting of surface modifiers, stabilizer / surface modifier orientation on particles, stabilizer / surface modifier densities on particles, stabilizer: drug ratio, drug concentration, particles formed It can affect size and polydispersity, as well as the morphology of the particles formed.

The drug (or salt thereof) can be present in any suitable amount in the solvent. In some embodiments, the drug (or salt thereof) is at least about 0.001%, at least about 0.01%, at least about 0.1%, at least about 0.5% of the solvent in% by weight or% by weight (weight: volume). , At least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 10%, at least about 12% , At least about 15%, at least about 20%, at least about 40%, at least about 60%, or at least about 80%. In some cases, the drug (or salt thereof) is about 100% or less, about 90% or less, about 80% or less, about 60% or less, about 40% or less, about 20% or less, about 20% or less of the solvent in the solvent. 15% or less, about 12% or less, about 10% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or It can be present in an amount of about 1% or less. Combinations in the range mentioned above (eg, about 20% or less of the solvent and at least about 1%) are also possible. In some embodiments, the drug is present in the above range (but weight: volume).

The ratio of stabilizer / surface modifier to the drug (or salt thereof) in the solvent can also be varied. In some embodiments, the ratio of stabilizer / surface modifier to the drug (or salt thereof) is at least 0.001: 1, at least 0.01: 1, at least 0.01: by weight, molar ratio, or weight: volume ratio. 1, at least 1: 1, at least 2: 1, at least 3: 1, at least 5: 1, at least 10: 1, at least 25: 1, at least 50: 1, at least 100: 1, or at least 500: 1. In some cases, the ratio of stabilizers / surface modifiers to pharmaceuticals (or salts thereof) is 1000: 1 or less, 500: 1 or less, 100: 1 or less, 75: 1 or less, by weight or molar ratio. It may be 50: 1 or less, 25: 1 or less, 10: 1 or less, 5: 1 or less, 3: 1 or less, 2: 1 or less, 1: 1 or less, or 0.1: 1 or less. Combinations of the ranges mentioned above (eg, at least 5: 1 and 50: 1 or less ratios) are also possible. Other ranges are possible.

Stabilizers / surface modifiers can be, for example, polymers or surfactants. Examples of polymers are suitable polymers for use in coatings, as described in more detail later. Non-limiting examples of surfactants include L-α-phosphatidylcholine (PC), 1,2-dipalmitoylphosphatidylcholine (DPPC), oleic acid, sorbitan trioleate, sorbitan monooleate, sorbitan monolaurate, monolauric acid. Polyoxyethylene sorbitan, polyoxyethylene sorbitan monooleate, natural lecithin, oleyl polyoxyethylene ether, stearyl polyoxyethylene ether, lauryl polyoxyethylene ether, block copolymer of oxyethylene and oxypropylene, synthetic lecithin, diethylene glycol dioleate , Tetrahydrofurfuryl oleate, ethyl oleate, isopropyl myristate, glyceryl monooleate, glyceryl monostearate, glyceryl monolithinol, cetyl alcohol, stearyl alcohol, polyethylene glycol 400, cetylpyridinium chloride, benzalconium chloride, olive oil Includes, glyceryl monolaurate, corn oil, cottonseed oil, and sunflower seed oil. Derivatives of the compounds mentioned above are also possible. Combinations of the compounds mentioned above with other compounds described herein can also be used as surface modifiers in the particles of the invention. As described herein, in some embodiments, the surface modifier can act as a stabilizer, surfactant, and / or emulsifier. In some embodiments, the surface modifier can aid in particle transport in the mucus.

In some embodiments, the stabilizer used for grinding forms a coating on the surface of the particles that makes the particles mucous permeable, whereas in other embodiments the stabilizer, particles are formed. Later, it can be replaced with one or more other surface modifiers. For example, in a set of methods, a first stabilizer / surface modifier can be used in the grinding step to coat the surface of the core particles, and then all or one of the first stabilizer / surface modifiers. The portions can be replaced with a second stabilizer / surface modifier to cover all or part of the surface of the core particles. In some cases, the second stabilizer / surface modifier can make the particles more mucous permeable than the first stabilizer / surface modifier. In some embodiments, core particles with coatings containing various surface modifiers can be formed.

Any suitable grinding medium can be used for grinding. In some embodiments, ceramic and / or polymeric materials and / or metals can be used. Examples of suitable materials are zirconium oxide, silicon carbide, silicon oxide, silicon nitride, zirconium silicate, yttrium oxide, glass, alumina, α-alumina, aluminum oxide, polystyrene, poly (methyl methacrylate), titanium, steel. Is included. The milling medium can have any suitable size. For example, the grinding medium can have an average diameter of at least about 0.1 mm, at least about 0.2 mm, at least about 0.5 mm, at least about 0.8 mm, at least about 1 mm, at least about 2 mm, or at least about 5 mm. In some cases, the grinding medium can have an average diameter of about 5 mm or less, about 2 mm or less, about 1 mm or less, about 0.8 mm or less, about 0.5 mm or less, or about 0.2 mm or less. Combinations in the range mentioned above (eg, average diameters of at least about 0.5 mm and about 1 mm or less) are also possible. Other ranges are possible.

Any suitable solvent can be used for grinding. The choice of solvent is, among other factors, the solid material to be ground (eg, pharmaceuticals), the individual type of stabilizer / surface modifier used (eg, one that can make the particles mucous permeable). May depend on factors such as the grinding material used. Suitable solvents may be those that do not substantially dissolve solid or ground materials, but dissolve stabilizers / surface modifiers to an appropriate degree. Non-limiting examples of solvents include pharmaceutical excipients, polymers, pharmaceuticals, salts, preservatives, viscosity regulators, isotonic regulators, flavor masking agents, antioxidants, pH regulators, and other pharmaceuticals. Examples may include water, buffer solutions, other aqueous solutions, alcohols (eg, ethanol, methanol, butanol), and mixtures thereof, which may optionally contain other components such as an excipient. In other embodiments, organic solvents can be used. The pharmaceutical product can have any suitable solubility in these or other solvents, eg, with respect to water solubility, or with respect to solubility in a coating solution, one or more of the above ranges of solubility.

In other embodiments, the core particles can be formed by sedimentation techniques. Precipitation techniques (eg, microsettling techniques, nanosettling techniques) can include forming a first solution containing a material (eg, a pharmaceutical product) that is substantially soluble in a solvent and a solvent used as a core. .. This solution can be added to a second solution containing another solvent in which the material is substantially insoluble, thereby forming multiple particles containing the material. In some cases, one or more surface modifiers, surfactants, materials, and / or bioactive agents can be present in the first and / or second solution. The coating can be formed during the process of settling the core (eg, the settling and coating steps can be performed substantially simultaneously). In another embodiment, the particles are first formed using a sedimentation technique and then the particles are coated with a surface modifier.

In some embodiments, sedimentation techniques can be used to form particles consisting of pharmaceutical salts (eg, nanocrystals). In general, the precipitation technique involves dissolving the material to be used as the wick in a solvent, and then the solution is a miscible antisolvent with or without excipients for forming the wick particles. Add to. This technique may be useful for preparing particles of pharmaceuticals that are soluble in aqueous solutions (eg, pharmaceuticals with relatively high water solubility). In some embodiments, pharmaceuticals with one or more charged or ionizable groups can interact with counterions (eg, cations or anions) to form salt complexes. For example, the pharmaceutical tenofovir (TFV) interacts very strongly with zinc cations via phosphonate groups and purine ring structures. This interaction with zinc can cause sedimentation of the TFV in the crystalline state, stabilizing the crystals with the coatings described herein and stopping agglutination.

Various counterions can be used to form salt complexes containing metals (eg, alkali metals, alkaline earth metals, and transition metals). Non-limiting examples of cationic counterions include zinc, calcium, aluminum, zinc, barium, magnesium, and copper. Non-limiting examples of anionic counterions include phosphate, carbonate, and fatty acids. The counterion may be, for example, monovalent, divalent, or trivalent. Other counterions are also known in the art and can be used in the embodiments described herein.

A variety of different acids can be used in the sedimentation method. In some embodiments, a suitable acid is deconic acid (deconic). Acid), caproic acid, mucic acid, octanic acid can be mentioned. In other embodiments, suitable acids include acetic acid, adipic acid, L-ascorbic acid, L-aspartic acid, capric acid (decanoic acid), carbonic acid, citric acid, fumaric acid, galactal acid, D-glucoheptic acid, D-gluconic acid, D-glucuronic acid, glutamic acid, glutaric acid, glycerophosphate, glycolic acid, horseuric acid, hydrochloric acid, DL-lactic acid, lauric acid, maleic acid, (-)-L-apple acid, palmitic acid, phosphoric acid , Sevacinic acid, stearic acid, succinic acid, sulfuric acid, (+)-L-tartrate acid, or thiocyan acid. In other embodiments, suitable acids include alginic acid, benzenesulfonic acid, benzoic acid, (+)-sulphonic acid, capric acid (octanoic acid), cyclamic acid, dodecylsulfate, ethane-1,2-disulfonic acid, Etan Sulfonic Acid, 2-Hydroxy Etan Sulfonic Acid, Gentisic Acid, 2-oxoglutaric Acid, Isobutyric Acid, Lactobionic Acid, Malonic Acid, Methan Sulfonic Acid, Naphthalene-1,5-Disulfonic Acid, Naphthalene-2-sulfonic Acid, 1- Hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, pamonic acid (embonic acid), propionic acid, (-)-L-pyroglutamic acid, or p-toluenesulfonic acid can be mentioned. In still other embodiments, suitable acids include 2,2-dichloroacetic acid, 4-acetamide-benzoic acid, (+)-kanfa-10-sulfonic acid, caproic acid (hexanoic acid), cinnamic acid, formic acid, Hydrobromic acid, DL-mandelic acid, nitrate, salicylic acid, 4-amino-salicylic acid, or undecyl acid (Undeca-10-enoic acid) can be mentioned. Mixtures of one or more such acids can also be used.

A variety of different bases can be used in the sedimentation method. In some embodiments, suitable bases include ammonia, L-arginine, calcium hydroxide, choline, N-methyl-glucamine, lysine, magnesium hydroxide, potassium hydroxide, or sodium hydroxide. In another embodiment, suitable bases include venetamine, benzathine, betaine, deanol, diethylamine, 2- (diethylamino) -ethanol, hydrababin, morpholine, 4- (2-hydroxyethyl)-, 1- (2-hydroxy). Ethyl) -pyrrolidine, or tromethamine can be mentioned. In other embodiments, suitable bases include diethanolamine (2,2'-iminobis (ethanol)), ethanolamine (2-aminoethanol), ethylenediamine, 1H-imidazole, piperazine, triethanolamine (2,2'). , 2''-Nitrilotris (ethanol)), or zinc hydroxide. Mixtures of one or more such bases can also be used.

Any suitable solvent can be used for sedimentation, including the solvents described herein that can be used for grinding. In one set of embodiments, aqueous solutions (eg, water, buffers, other aqueous solutions), alcohols (eg, water, buffers, other aqueous solutions), which may optionally contain other components such as pharmaceutical excipients, polymers, and pharmaceuticals. , Ethanol, methanol, butanol), and mixtures thereof.

In the precipitation method, the salt can have lower water solubility (or solubility in a solvent containing the salt) than the non-salt form of the drug. The water solubility (or solubility in a solvent) of the salt is at 25 ° C, for example, about 5 mg / mL or less, about 2 mg / mL or less, about 1 mg / mL or less, about 0.5 mg / mL or less, about 0.1 mg / mL or less, about 0.05 mg / mL or less, or about 0.01 mg / mL or less, about 1 μg / mL or less, about 0.1 μg / mL or less, about 0.01 μg / mL or less, about 1 ng / mL or less, about 0.1 ng / mL or less , Or about 0.01 ng / mL or less. In some embodiments, the salt is at least about 1 pg / mL, at least about 10 pg / mL, at least about 0.1 ng / mL, at least about 1 ng / mL, at least about 10 ng / mL, at least about 0.1 μg / mL, at least about about. 1 μg / mL, at least about 5 μg / mL, at least about 0.01 mg / mL, at least about 0.05 mg / mL, at least about 0.1 mg / mL, at least about 0.5 mg / mL, at least about 1.0 mg / mL, at least about 2 mg / mL Can have water solubility (or solubility in a solvent). Combinations in the range mentioned above (eg, at least about 0.001 mg / mL and less than about 1 mg / mL water soluble (or soluble in solvent)) are also possible. Other ranges are possible. The salt can have water solubility in these or other ranges at any point in the pH range (eg, pH 1 to pH 14).

In some embodiments, the solvent used for precipitation comprises one or more surface modifiers as described herein, one or more surface modifiers as the particles settle from the solution. A coating consisting of can be formed around the particles. The surface modifier is at least about 0.001%, at least about 0.005%, at least about 0.01%, at least about 0.05%, at least about. It can be present at a concentration of 0.1%, at least about 0.5%, at least about 1%, or at least about 5%. In some cases, the surface modifier is in solvent in% by weight / volume of about 5% or less, about 1% or less, about 0.5% or less, about 0.1% or less, about 0.05% or less, about 0.01% or less. , Or at a concentration of about 0.005% or less. Combinations in the range mentioned above (eg, concentrations of at least about 0.01 (weight / volume)% and about 1 (weight / volume)% or less) are also possible. Other ranges are possible.

Another typical method of forming core particles involves freeze-drying techniques. In this technique, the medicinal product or a salt thereof can be dissolved in an aqueous solution which may contain a surface modifier. Counterions can be added to this solution and the solution can be snap frozen immediately and lyophilized. The dry powder can be reconstituted at the desired concentration in a suitable solvent (eg, an aqueous solution such as water).

Counterions can be added to the lyophilization solvent in any suitable range. In some cases, the ratio of the counterion to a drug (eg, salt) is at least 0.1: 1, at least 1: 1, at least 2: 1, at least 3: 1, at least 5: 1 in weight or molar ratio. , At least 10: 1, at least 25: 1, at least 50: 1, or at least 100: 1. In some cases, the ratio of the counterion to a drug (eg, salt) is 100: 1 or less, 75: 1 or less, 50: 1 or less, 25: 1 or less, 10: 1 or less in weight or molar ratio. , 5: 1 or less, 3: 1 or less, 2: 1 or less, 1: 1 or less, or 0.1: 1 or less. Combinations of the ranges mentioned above (eg, at least 5: 1 and 50: 1 or less ratios) are also possible. Other ranges are possible.

If the surface modifier is present in the solvent prior to lyophilization, the surface modifier is at least about 0.001%, at least about 0.005%, at least in an aqueous solution at any suitable concentration, eg, by weight / volume%. It can be present at a concentration of about 0.01%, at least about 0.05%, at least about 0.1%, at least about 0.5%, at least about 1%, or at least about 5%. In some examples, the surface modifier is in solvent in% by weight / volume of about 5% or less, about 1% or less, about 0.5% or less, about 0.1% or less, about 0.05% or less, about 0.01% or less. , Or at a concentration of about 0.005% or less. Combinations in the range mentioned above (eg, concentrations of at least about 0.01 (weight / volume)% and about 1 (weight / volume)% or less) are also possible. Other ranges are possible.

The concentration of the surface modifier present in the solvent may or may not exceed the critical micelle concentration (CMC) of the surface modifier and depends on the individual surface modifier used. For example, as described in the Examples section, F127 concentrations both above (1%) and below CMC (0.08%) of F127 are used to coat stable nanocrystalline particles of the pharmaceutical tenofovir. be able to. However, nanocrystalline particles of acyclovir monophosphate are more sensitive to detergent concentrations, and stable nanocrystalline particles form only when utilizing F127 concentrations below CMC (about 0.1%). We were able to.

In other embodiments, stable particles can be formed by adding excess counterions to a solution containing a pharmaceutical product. The sediment can then be washed by various methods such as centrifugation. The resulting slurry can be treated with ultrasonic waves. One or more surface modifiers can be added to stabilize the resulting particles.

Other methods of forming pharmaceutical particles are also possible. So-called top-down techniques include, for example, grinding techniques and high pressure homogenization. In high pressure homogenization, a suspension of material to be used as a core is pushed under pressure through a gap, valve, or opening to reduce the particle size. So-called bottom-up techniques include, for example, precipitation, emulsification (material to be used as a core dissolved in a solvent is added to an immiscible antisolvent with or without excipients to add core particles. Includes (forming) and spray drying (spraying a solution of the material to be used as the core into the antisolvent of the gas phase to form core particles).

Combinations of the methods described herein with other methods are also possible. For example, in some embodiments, a core made of a pharmaceutical product is first formed by sedimentation, and then the size of the core is further reduced in a grinding step.

Following the formation of the particles of pharmaceuticals, the particles can optionally be exposed to a solution containing a (second) surface modifier capable of binding and / or coating the particles. In embodiments where the pharmaceutical product already comprises a coating consisting of a first surface modifier, the second surface modifier may be replaced in whole or in part with a second stabilizer / surface modifier to cover all or part of the particle surface. can do. In some cases, the second surface modifier can make the particles more mucous permeable than the first surface modifier. In other embodiments, it is possible to form particles with a coating (eg, in a single or multi-layered state) containing a variety of surface modifiers. In other embodiments, particles can be formed with a variety of coatings (eg, each coating optionally comprises a different surface modifier). In some cases, the coating exists in the form of a single layer consisting of a surface modifier. Other arrangements are possible.

In any of the methods described herein, the particles are placed in solution with a surface modifier for at least about 1 minute, at least about 2 minutes, at least about 5 minutes, at least about 10 minutes, at least about 15 minutes, at least about 20 minutes. Particles can be coated by incubating for minutes, at least about 30 minutes, at least about 60 minutes, or longer. In some cases, the incubation can be done in about 10 hours or less, about 5 hours or less, or about 60 minutes or less. Combinations in the range mentioned above (eg, incubation time of 60 minutes or less and at least 2 minutes) are also possible.

Particle Coating As shown in the illustrated embodiment of FIG. 1, the core (16) can be surrounded by a coating (20) containing one or more surface modifiers. In some embodiments, the coating is formed of one or more surface modifiers, or other molecules, placed on the surface of the core. The individual chemical composition and / or components of the coating and surface modifier can be selected to impart specific functionality, eg, enhanced transport within the mucosal barrier, to the particles.

It should be understood that the coating surrounding the wick may have embodiments that completely enclose the wick, but it does not necessarily have to completely enclose the wick. For example, the coating covers at least about 10%, at least about 30%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 99% of the surface area of the core. Can be surrounded. In some cases, the coating substantially surrounds the core. In other cases, the coating completely surrounds the core. In other embodiments, the coating surrounds about 100% or less, about 90% or less, about 80% or less, about 70% or less, about 60% or less, or about 50% or less of the surface area of the core. Combinations in the range mentioned above (eg, surrounding more than 80% and less than 100% of the surface area of the core) are also possible.

The components of the coating can be evenly distributed over the entire surface of the core in some cases and non-uniformly in others. For example, the coating can, in some cases, include parts (eg, holes) that are completely free of material. Allows penetration and / or transport of certain molecules and components into and / or from the coating, if desired, but prevents penetration and / or transport of other molecules or components into or from the coating. The coating can be designed as such. The ability of a particular molecule to penetrate and / or be transported into and / or across a coating is, for example, the packing density of the surface modifier forming the coating, and the chemical and physical properties of the components forming the coating. May depend on. As described herein, the coating can include one material layer or, in some embodiments, multiple material layers. There can be a single surface modifier or multiple surface modifiers.

The coating of particles can have any suitable thickness. For example, the coating has an average thickness of at least about 1 nm, at least about 5 nm, at least about 10 nm, at least about 30 nm, at least about 50 nm, at least about 100 nm, at least about 200 nm, at least about 500 nm, at least about 1 μm, or at least about 5 μm. Can have. In some cases, the average thickness of the coating is about 5 μm or less, about 1 μm or less, about 500 nm or less, about 200 nm or less, about 100 nm or less, about 50 nm or less, about 30 nm or less, about 10 nm or less, or about 5 nm or less. be. Combinations in the range mentioned above (eg, average thickness of at least about 1 nm and about 100 nm or less) are also possible. Other ranges are possible. For particles with multiple coatings, each coating layer can have one of the above thicknesses.

In some embodiments, the compositions and methods described herein are such that the core particles are coated with a hydrophilic surface modification without the need for covalent connection of the surface modification to the core surface. Can be made possible. In some such embodiments, the core having a hydrophobic surface is coated with the polymers described herein, whereby a plurality on the core surface without substantially altering the properties of the core itself. The surface modification part of can be present. For example, the surface modifier can be adsorbed on the outer surface of the core particles. However, in other embodiments, the surface modifier is covalently linked to the core particles.

In certain embodiments where the surface modifier is adsorbed on the surface of the wick, the surface modifier is an alternative molecule of the surface modifier in solution and optionally other components (eg, in the composition / formulation). ) Can exist in equilibrium. In some cases, the adsorbed surface modifier can be present on the surface of the wick at the densities described herein. Since the surface modifier exists in equilibrium with other components in the solution, the density may be an average density.

The particle coating and / or surface modifiers described herein can include any suitable material, such as, for example, hydrophobic materials, hydrophilic materials, and / or amphipathic materials. In certain embodiments, the polymer comprises a synthetic polymer (ie, a polymer that is not produced in nature). In other embodiments, the polymer is a natural polymer (eg, protein, polysaccharide, rubber). In certain embodiments, the polymer is a polymer having surface activity. In certain embodiments, the polymer is a nonionic polymer. In certain embodiments, the polymer is a nonionic block copolymer. In some embodiments, the polymer may be a diblock copolymer, a triblock copolymer, for example, one block of which is a hydrophobic polymer and another of which is a hydrophilic polymer. The polymer may be charged or non-charged.

In some embodiments, the particles described herein comprise a coating comprising a block copolymer having a relatively hydrophilic block and a relatively hydrophobic block. In some cases, the hydrophilic block can be substantially present on the outer surface of the particles. For example, hydrophilic blocks can form most of the outer surface of the coating and can help stabilize the particles in an aqueous solution containing the particles. It is substantially present inside the coating and / or on the surface of the core particles, which can facilitate, for example, the coating to adhere to the core. In some cases, the coating comprises a surface modifier comprising a triblock copolymer, wherein the triblock copolymer comprises a hydrophilic block-hydrophobic block-hydrophilic block arrangement.

The molecular weights of the hydrophilic and hydrophobic blocks of the triblock copolymer can be selected, respectively, to reduce the mucous adhesion of the core and to ensure sufficient association of the triblock copolymer with the core. As described herein, the molecular weight of the hydrophobic block of the triblock copolymer increases the probability that the triblock copolymer will remain adhered to the core due to sufficient association between the triblock copolymer and the core. Can be selected for. Surprisingly, in certain embodiments, if the molecular weight of the hydrophobic block of the triblock copolymer is too small (eg, about 2 kDa or less), sufficient adhesion between the hydrophobic core and the triblock copolymer cannot be expected. Therefore, it has been found that particles with such hydrophobic blocks may not exhibit a sufficiently reduced mucous stickiness.

In certain embodiments, the hydrophobic block of the triblock copolymer (eg, the PPO block of the triblock copolymer PEG-PPO-PEG, where the PEG block is compatible with the PEO block) has a molecular weight of at least about 2 kDa, at least about about. 3 kDa, at least about 4 kDa, at least about 5 kDa, at least about 6 kDa, at least about 10 kDa, at least about 20 kDa, or at least about 50 kDa. In some embodiments, the molecular weight of the hydrophobic block is about 100 kDa or less, about 80 kDa or less, about 50 kDa or less, about 20 kDa or less, about 15 kDa or less, about 13 kDa or less, about 12 kDa or less, about 10 kDa or less, about 8 kDa or less. , Or about 6 kDa or less. Combinations in the range mentioned above (eg, at least about 3 kDa and about 15 kDa or less) are also possible. Other ranges are possible.

It has also been found that in certain embodiments, a sufficient amount of hydrophilic block (as a function of the total weight of the polymer) is required for the particles to exhibit a sufficiently reduced mucous adhesion. For example, in certain embodiments, hydrophilic blocks constituting at least about 15% by weight (eg, at least about 20% by weight, at least about 25% by weight, or at least about 30% by weight) of triblock copolymers mucous the particles. Permeability, while mucous stickiness, was commonly observed with particles in which the weight ratio of hydrophilic blocks was below this limit. In some embodiments, the hydrophilic block of the triblock copolymer is at least about 15% by weight, at least about 20% by weight, at least about 25% by weight, at least about 30% by weight, at least about 35% by weight. , At least about 40% by weight, at least about 45% by weight, at least about 50% by weight, at least about 55% by weight, at least about 60% by weight, at least about 65% by weight, or at least about 70% by weight. In some embodiments, the hydrophilic block of the triblock copolymer is about 90% by weight or less, about 80% by weight or less, about 60% by weight or less, about 50% by weight or less, or about 40% by weight of the triblock copolymer. It consists of the following. Combinations in the range mentioned above (eg, at least about 30% by weight and about 80% by weight or less) are also possible. Other ranges are possible.

In some embodiments, the molecular weight of the hydrophilic block (eg, the PEG (or PEO) block of the triblock copolymer PEG-PPO-PEG, where the PEG block is compatible with the PEO block) is at least about 0.05 kDa. , At least about 0.1 kDa, at least about 0.2 kDa, at least about 0.3 kDa, at least about 0.4 kDa, at least about 0.5 kDa, at least about 1 kDa, at least about 2 kDa, at least about 3 kDa, at least about 4 kDa, at least about 5 kDa, at least about 6 kDa, It can be at least about 8 kDa, at least about 10 kDa, at least about 20 kDa, or at least about 50 kDa. In certain embodiments, the molecular weight of the hydrophilic block is about 100 kDa or less, about 80 kDa or less, about 50 kDa or less, about 20 kDa or less, about 15 kDa or less, about 10 kDa or less, about 9 kDa or less, about 8 kDa or less, about 7 kDa or less, It can be about 6 kDa or less, about 5 kDa or less, about 3 kDa or less, about 2 kDa or less, or about 1 kDa or less. Combinations in the range mentioned above (eg, at least about 0.1 kDa and about 3 kDa or less) are also possible. Other ranges are possible. In embodiments where the two hydrophilic blocks are located on either side of the hydrophobic block, the molecular weights of the two hydrophilic blocks may be substantially the same or different.

In certain embodiments, the polymer of the surface modifier comprises a polyether moiety. In certain embodiments, the polymer comprises a polyalkyl ether moiety. In certain embodiments, the polymer comprises a tail that is polyethylene glycol. In certain embodiments, the polymer comprises a central portion that is polypropylene glycol. In certain embodiments, the polymer comprises polybutylene glycol as a central moiety. In certain embodiments, the polymer comprises polypentylene glycol as a central portion. In certain embodiments, the polymer comprises polyhexylene glycol as a central portion. In certain embodiments, the polymer is a triblock copolymer consisting of one of the polymers described herein. As disclosed herein, any enumeration of PEG can be replaced with polyethylene oxide (PEO) and any enumeration of PEO can be replaced with PEG.

In certain embodiments, the polymer is a triblock copolymer consisting of a polyalkyl ether (eg, polyethylene glycol, polypropylene glycol) and another polymer. In certain embodiments, the polymer is a triblock copolymer consisting of a polyalkyl ether and another polyalkyl ether. In certain embodiments, the polymer is a triblock copolymer consisting of polyethylene glycol and another polyalkyl ether. In certain embodiments, the polymer is a triblock copolymer consisting of polypropylene glycol and another polyalkyl ether. In certain embodiments, the polymer is a triblock copolymer containing at least one unit of polyalkyl ether. In certain embodiments, the polymer is a triblock copolymer consisting of two different polyalkyl ethers. In certain embodiments, the polymer is a triblock copolymer containing polyethylene glycol units. In certain embodiments, the polymer is a triblock copolymer containing polypropylene glycol units. In certain embodiments, the polymer is a triblock copolymer consisting of more hydrophobic units with two more hydrophilic units located on either side. In certain embodiments, the hydrophilic unit is a polymer of the same type. In certain embodiments, the polymer comprises a polypropylene glycol unit in which two more hydrophilic units are located on either side. In certain embodiments, the polymer comprises two polyethylene glycol units with more hydrophobic units located on either side. In certain embodiments, the polymer is a triblock copolymer having polypropylene glycol units with two polyethylene glycol units located on either side. The molecular weights of the two blocks located on either side of the central block may be substantially the same or different.

In certain embodiments, the polymer has the formula:<chemistry><img file="JP2022017589A_D0001.tif" /></chemistry>In the equation, n is an integer of 2 to 1140 (including both ends), and m is an integer of 2 to 1730 (including both ends). In certain embodiments, n is an integer from 10 to 170 (including both ends). In certain embodiments, m is an integer from 5 to 70 (including both ends). In certain embodiments, n is at least twice m, three times m, or four times m.

In certain embodiments, the coating comprises a surface modifier comprising a poly (ethylene glycol) -poly (propylene oxide) -poly (ethylene glycol) triblock copolymer (hereinafter "PEG-PPO-PEG triblock copolymer"). include. As described herein, in some embodiments, PEG blocks can be interchanged with PEO blocks. The molecular weight of the PEG (or PEO) and PPO moieties of the PEG-PPO-PEG triblock copolymer can be selected to reduce mucoadhesion of the particles, as described herein. Without being bound by theory, particles with coatings containing PEG-PPO-PEG triblock copolymers are controlled by exposing multiple PEG (or PEO) moieties on the particle surface, at least in part. Mucus adhesion can be reduced as compared with particles. The PPO moiety can adhere to the core surface (eg, if the core surface is hydrophobic), thus allowing strong association between the core and the triblock copolymer. In some cases, PEG-PPO-PEG triblock copolymers associate with the core via non-covalent interactions. For comparison purposes, the control particles may be, for example, carboxylate-modified polystyrene particles of similar size to the coated particles.

In certain embodiments, surface modifiers include polymers containing poloxamers with the trade name Pluronic®. Pluronic® polymers that may be useful in the embodiments described herein are, but are not limited to, F127, F38, F108, F68, F77, F87, F88, F98, L101, L121, Examples include L31, L35, L43, L44, L61, L62, L64, L81, L92, N3, P103, P104, P105, P123, P65, P84 and P85.

Table 1 shows examples of the molecular weights of specific Pluronic® molecules.

<tables><img file="JP2022017589A_D0002.tif" /></tables>

Other ranges are possible and may be useful in the particular embodiments described herein, but in some embodiments the hydrophobic block of the PEG-PPO-PEG triblock copolymer is. Having one of the molecular weights (eg, at least about 3 kDa and about 15 kDa or less), the hydrophilic blocks together are one weight percent of the range (eg, at least about 15% by weight, at least about about) relative to the polymer. 20% by weight at least about 25% by weight, or at least about 30% by weight, and about 80% by weight or less). Specific Pluronic® polymers included within these criteria include, for example, F127, F108, P105 and P103. Surprisingly, as described in more detail in the examples, these particular Pluronic® polymers extend particles beyond other Pluronic® polymers under test that are not included within this standard. It was found to be mucus permeable. In addition, other agents that did not make the particles mucopermeable include specific polymers such as polyvinylpyrrolidone (PVP / Kollidon) polyvinyl alcohol-polyethylene glycol graft copolymer (Kollicoat IR), hydroxypropylmethylcellulose (Methocel); Tween20, Oligomers such as Tween80, Soltor HS15, Triton X100, Tyroxapol, Cremohole RH40; small molecules such as Span20, Span80, octylglucoside, cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS).

Much of the description herein relates to coatings containing a hydrophilic block-hydrophobic block-hydrophilic block arrangement (eg, PEG-PPO-PEG triblock copolymer), but coatings are limited to this arrangement. Please be aware that this is not the case, and that other arrangements and materials are possible. For example, the particles include a single coating, but in other embodiments, the particles include more than one coating (eg, at least two, three, four, five, or more coatings). Each coating can be formed from a mucous permeable material or does not necessarily include a mucous permeable material. In some cases, the intermediate coating (ie, the coating between the core surface and the outer coating) can include a polymer that facilitates the attachment of the outer coating to the core surface. In many embodiments, the outer coating of the particles comprises a polymer containing a material that facilitates the transport of the particles in the mucus.

Thus, the coating (eg, inner coating, intermediate coating, and / or outer coating) can include any suitable polymer. In some cases, the polymer may be biocompatible and / or biodegradable. In some cases, the polymeric material can include more than one polymer (eg, at least 2, 3, 4, 5 or more polymers). In some cases, the polymer may be a random copolymer or a block copolymer as described herein (eg, diblock copolymer, triblock copolymer).

Non-limiting examples of suitable polymers include polyamine, polyether, polyamide, polyester, polycarbamate, polyurea, polycarbonate, polystyrene, polyimide, polysulfone, polyurethane, polyacetylene, polyethylene, polyethyleneimine, polyisocyanate, polyacrylate, polymethacrylate. , Polyacrylonitrile, and polyarylate. Non-limiting examples of specific polymers include poly (caprolactone) (PCL), ethylene vinyl acetate polymer (EVA), poly (lactic acid) (PLA), poly (L-lactic acid) (PLLA), poly (glycolic acid). ) (PGA), Poly (lactic acid-co-glycolic acid) (PLGA), Poly (L-lactic acid-co-glycolic acid) (PLLGA), Poly (D, L-lactide) (PDLA), Poly (L-lactide) ) (PLLA), Poly (D, L-Lactide-co-Caprolactone), Poly (D, L-Lactide-co-Caprolactone-co-Glycolide), Poly (D, L-Lactide-co-PEO-co-D) , L-lactide), poly (D, L-lactide-co-PPO-co-D,

The molecular weight of the polymer may vary. In some embodiments, the molecular weights are at least about 0.5 kDa, at least about 1 kDa, at least about 2 kDa, at least about 3 kDa, at least about 4 kDa, at least about 5 kDa, at least about 6 kDa, at least about 8 kDa, at least about 10 kDa, at least about 12 kDa. , At least about 15 kDa, at least about 20 kDa, at least about 30 kDa, at least about 40 kDa, or at least about 50 kDa. In some embodiments, the molecular weight is about 50 kDa or less, about 40 kDa or less, about 30 kDa or less, about 20 kDa or less, about 12 kDa or less, about 10 kDa or less, about 8 kDa or less, about 6 kDa or less, about 5 kDa or less, or about 4 kDa or less. It's fine. Combinations in the range mentioned above (eg, molecular weights of at least about 2 kDa and about 15 kDa or less) are also possible. Other ranges are possible. The molecular weight can be measured using any known technique such as light scattering, gel permeation chromatography and the like. Other methods are also known in the art.

In certain embodiments, the polymer is biocompatible, i.e., the polymer does not typically induce a detrimental response when inserted or injected into a living subject; for example, it is a significant inflammation. And / or does not include acute rejection of the polymer by the immune system, eg, through a T cell-mediated response. Of course, "biocompatibility" is a relative term, and it will be recognized that even for polymers that are highly compatible with living tissue, some degree of immune response is expected. However, as used herein, "biocompatibility" is associated with acute rejection of the material by at least a portion of the immune system, i.e., non-biocompatible material implanted in the subject is the immune system. Induces an immune response in a subject that is too severe to adequately control the rejection of the material by and often to the extent that the material must be removed from the subject. One simple test to measure biocompatibility is to expose the polymer to cells in vitro: the biocompatible polymer is at moderate concentrations, eg about 50 μg / 10⁶It is a polymer that typically does not result in significant cell death at cell concentration. For example, biocompatible polymers can cause cell death when exposed to cells such as fibroblasts or epithelial cells, even if they are phagocytosed or otherwise taken up by such cells. Less than%. In some embodiments, the substance causes less than 20% cell death by adding it to cells in vitro, and it is not desirable to administer them in vivo for inflammation or other such. If it does not induce adverse effects, it is "biocompatible".

In certain embodiments, the biocompatible polymer may be biodegradable, i.e., chemically and / or biologically (ie, in a physiological environment such as when the polymer is introduced into the body or cells). It can be degraded (eg, by cellular mechanism or by hydrolysis). For example, the polymer may spontaneously hydrolyze upon exposure to water (eg, within the subject) and / or the polymer may decompose upon exposure to heat (eg, a temperature of about 37 ° C). Can be done. Polymer degradation can occur at various rates, depending on the polymer or copolymer used. For example, the half-life of a polymer (the time it takes for 50% of a polymer to decompose into monomers and / or other non-polymeric moieties) can be in the order of days, weeks, months, or years, depending on the polymer. It's fine. Polymers can be biologically degraded, for example, by enzymatic activity or cellular mechanism, in some cases, for example, via exposure to lysozyme (eg, having a relatively low pH). In some cases, the polymer can be reused or processed by the monomer and / or the cell without significant toxic effects on the cell (ie, when the component is added to the cell in vitro, about 20% of the cells die. May be decomposed into other non-polymeric moieties (less than). For example, polylactide can be hydrolyzed to form lactic acid, polyglycolide can be hydrolyzed to form glycolic acid, and so on.

Examples of biodegradable polymers include, but are not limited to, poly (ethylene glycol) -poly (propylene oxide) -poly (ethylene glycol) triblock copolymers, poly (lactide) (or poly (lactic acid)), poly (glycolide). ), (Or poly (glycolic acid)), poly (orthoester), poly (caprolactone), polylysine, poly (ethyleneimine), poly (acrylic acid), poly (urethane), poly (amphilide), poly (ester) , Poly (trimethylene carbonate), poly (ethyleneimine), poly (acrylic acid), poly (urethane), poly (β-aminoester), etc., and copolymers or derivatives of these and / or other polymers, such as poly. (Lactide-co-glycolide) (PLGA) is included.

In certain embodiments, the polymer may be biodegradable within the time period allowed for the desired application. In certain embodiments, such as in vivo therapy, such degradation is usually about 5 years, 1 year, 6 months with exposure to a physiological solution at a temperature of 25-37 ° C and a pH of 6-8. , 3 months, 1 month, 15 days, 5 days, 3 days or less, or even less than 1 day (eg 1-4 hours, 4-8 hours, 4-24 hours, 1-24 hours) It happens in. In other embodiments, the polymer decomposes over a period of about 1 hour to several weeks, depending on the desired application.

The coatings and particles described herein can include polymers, but in some embodiments, the particles described herein are hydrophobic materials that are not polymers (eg, non-polymers) and are not pharmaceuticals. including. For example, in some embodiments, all or part of the particles can be coated with a passivation layer. Non-limiting examples of non-polymeric materials include certain metals, waxes, and organic materials (eg, organic silanes, perfluorinated or fluorinated organic materials).

Particles with Reduced Mucosal Adhesion As described herein, in some embodiments, the method comprises locating a material, such as particles, whose mucosal adhesiveness is desired to be reduced. Materials that require increased diffusivity in mucus may be, for example, hydrophobic, have many hydrogen bond donors or acceptors, and / or be highly charged. In some cases, the material may include crystalline or amorphous solid materials. Materials that may serve as wicks may be coated with the appropriate polymers described herein, thereby forming particles with multiple surface modification moieties on the surface, reducing mucoidity. Can bring. Particles herein described as having reduced mucous stickiness are, otherwise, increased transport in mucus, mobile in mucus, or mucous permeability (ie, mucus permeability). Characterized as (mucus-permeable particles), it means that the particles are transported in the mucus faster than the (negative) control particles. The (negative) control particles may be particles known to be mucous sticky, such as unmodified particles or cores not coated with the coatings described herein, such as 200 nm carboxylated polystyrene particles.

In certain embodiments, the method of the invention is a state of a formulation configured for delivery (eg, topical delivery) of a pharmaceutical composition or formulation consisting of a modified substance, eg, to a mucous or mucosal surface of interest. Includes preparation in. Pharmaceutical compositions with surface-modified moieties can, for example, be delivered to the mucosal surface of the subject, can pass through the mucosal barrier in the subject, and / or have mucosal surfaces because of reduced mucous stickiness. It is possible to prolong the retention of particles in and / or increase the uniform distribution. As is known to those skilled in the art, mucus is a viscoelastic sticky substance that captures most foreign particles. The captured particles are unable to reach the underlying epithelium and / or are rapidly eliminated by the mucus elimination mechanism. In order for the particles to reach the underlying epithelium and / or for their long stay in the mucosal tissue, the particles must rapidly penetrate the mucous secretions and / or circumvent the mucous clearance mechanism. It doesn't become. If the particles do not substantially adhere to the mucus, the particles can diffuse into the interstitial fluid between the mucin fibers and reach the underlying epithelium and / or are not eliminated by the mucus exclusion mechanism. Therefore, modifying a mucous sticky material (eg, a pharmaceutical that is hydrophobic) with a material to reduce the mucous stickiness of the particles can result in efficient delivery of the particles to the underlying epithelium and / or mucosal surface. Can allow long retention of particles in.

Moreover, in some embodiments, the particles described herein with reduced mucous stickiness promote better distribution of particles on the tissue surface as compared to particles that are more mucous sticky. And / or prolong its presence on the tissue surface. For example, in some cases, luminal spaces such as the gastrointestinal tract are surrounded by a mucus-covered surface. Mucous sticky particles delivered to such spaces are typically removed from the luminal space and mucous-covered surfaces by the body's natural exclusion mechanism. The particles described herein with reduced mucous stickiness can remain in the luminal space for a relatively longer period of time compared to mucous sticky particles. This long presence can prevent or reduce the exclusion of particles and / or allow better distribution of the particles on the tissue surface. Long presence can also affect particle transport in the luminal space, for example particles can be distributed in the mucous layer and reach the underlying epithelium.

In certain embodiments, the polymer-coated material (eg, core) described herein passes through a mucous or mucosal barrier in the subject and / or exhibits long retention of particles on the mucosal surface. And / or can increase the uniform distribution of particles, for example, such substances are more gradual (at least 2-fold, 5-fold, 10-fold) from the subject's body compared to (negative) control particles. , And even more gradually than at least 20 times). The (negative) control particles may be particles known to be mucous sticky, such as unmodified particles or cores not coated with the coatings described herein, such as 200 nm carboxylated polystyrene particles. ..

In certain embodiments, the particles described herein have a particular relative velocity &lt;V as defined as:_{mean mean}&gt;_relHave:<math num="1"><img file="JP2022017589A_D0003.tif" /></math> In the formula, &lt;V_{mean mean}&gt; Is the average velocity of all average trajectories, V_{mean mean}Is the velocity of the individual particles averaged over its trajectory, the sample is the particle of interest, the negative control is 200 nm carboxylated polystyrene particles, and the positive control is dense with 2 kDa-5 kDa PEG. 200 nm polystyrene particles PEGylated to.

Relative velocities can be measured by a variety of particle tracking techniques. For example, using a fluorescence microscope equipped with a CCD camera, a time of 66.7 ms for each type of particle, ie sample, negative control, and positive control, from several regions within each sample under 100X magnification. It can record 15 moving images (15 frames / sec) with a resolution. Samples, negative and positive controls may be fluorescent particles for observing trajectories. Alternatively, the non-fluorescent particles can be coated with a fluorescent molecule, a surface agent with a fluorescent tag, or a polymer with a fluorescent tag. Modern image processing software (eg Image Pro or MetaMorph) can be used to measure individual trajectories (50 frames) of various particles over a time scale of at least 3.335 seconds.

In some embodiments, the particles described herein are about 0.3 or more, about 0.4 or more, about 0.5 or more, about 0.6 or more, about 0.7 or more, about 0.8 or more, about 0.9 or more, about 1.0 in mucus. It has a relative velocity of about 1.1 or more, about 1.2 or more, about 1.3 or more, about 1.4 or more, about 1.5 or more, about 1.6 or more, about 1.7 or more, about 1.8 or more, about 1.9 or more, or about 2.0 or more. In some embodiments, the particles described herein are about 10.0 or less, about 8.0 or less, about 6.0 or less, about 4.0 or less, about 3.0 or less, about 2.0 or less, about 1.9 or less, about 1.8 in mucus. Relative velocity of about 1.7 or less, about 1.6 or less, about 1.5 or less, about 1.4 or less, about 1.3 or less, about 1.2 or less, about 1.1 or less, about 1.0 or less, about 0.9 or less, about 0.8 or less, or about 1.7 or less. Have. Combinations in the range mentioned above (eg, relative velocities of about 0.5 or more and about 6.0 or less) are also possible. Other ranges are possible. The mucus may be, for example, human cervical mucus.

In certain embodiments, the particles described herein are control particles or corresponding particles (eg, unmodified and / or uncoated with the coatings described herein) in a mucous or mucosal barrier. It can diffuse at a higher rate or diffusivity than particles). In some cases, the particles described herein have a mucous or mucosal barrier at least about 10-fold, 20-fold, 30-fold, 50-fold, 100-fold, 200-fold, compared to control or corresponding particles. It can pass at speeds or diffusion rates that are 500x, 1000x, 2000x, 5000x, 10000x, or higher. In some cases, the particles described herein have a mucous or mucosal barrier of about 10,000 times or less, about 5000 times or less, about 2000 times or less, about 1000 times or less, as compared to control particles or corresponding particles. It can pass at a rate or diffusion rate of about 500 times or less, about 200 times or less, about 100 times or less, about 50 times or less, about 30 times or less, about 20 times or less, or about 10 times or less. Combinations in the range mentioned above are also possible (eg, at least about 10 times and about 1000 times less than the control or corresponding particles). Other ranges are possible.

For the purposes of comparison described herein, the corresponding particles are about the same size, shape, and / or density as the test particles, but lack a coating that makes the test particles mobile in the mucus. In some cases, measurements are based on a time scale of about 1 second, about 0.5 seconds, about 2 seconds, about 5 seconds, or about 10 seconds. Those of skill in the art will know how to measure the square geometric mean displacement and velocity or diffusivity.

In addition, the particles described herein are at least about 10x, 20x, 30x, 50x, 100x, 200x, 500x, 1000x, 2000x, 5000x compared to the corresponding or control particles. It can pass through mucous or mucosal barriers with square geometric mean displacements of fold, 10,000 times, or more. In some cases, the particles described herein are about 10,000 times or less, about 5000 times or less, about 2000 times or less, about 1000 times or less, about 500 times or less, about, compared to control particles or corresponding particles. It can pass through mucous or mucosal barriers with a square geometric mean displacement of 200 times or less, about 100 times or less, about 50 times or less, about 30 times or less, about 20 times or less, or about 10 times or less. Combinations in the range mentioned above are also possible (eg, at least about 10 times and about 1000 times less than the control or corresponding particles). Other ranges are possible.

In some embodiments, the particles described herein diffuse in the mucosal barrier at a rate at which the particles can diffuse in water or at a rate approximately equal to the diffusion rate. In some cases, the particles described herein are about 1/2 or less, about 1/4 or less, about 1/8 or less, about 1/16 of the diffusivity of the particles diffusing in water under the same conditions. Below, about 1/32 or less, about 1/50 or less, about 1/100 or less, about 1/200 or less, about 1/300 or less, about 1/400 or less, about 1/500 or less, about 1/600 or less, Mucosa at a rate or diffusion rate of about 1/700 or less, about 1/800 or less, about 1/900 or less, about 1/1000 or less, about 1/2000 or less, about 1/5000 or less, about 1 / 10,000 or less. You can pass through the barrier. In some cases, the particles described herein are about 1 / 10,000 or more, about 1/5000 or more, about 1/2000 or more, about 1/1000 of the diffusivity of the particles diffusing in water under the same conditions. Above, about 1/900 or more, about 1/800 or more, about 1/700 or more, about 1/600 or more, about 1/500 or more, about 1/400 or more, about 1/300 or more, about 1/200 or more, At a rate or diffusion rate that is about 1/100 or more, about 1/50 or more, about 1/32 or more, about 1/16 or more, about 1/8 or more, about 1/4 or more, or about 1/2 or more. It can cross the mucosal barrier. Combinations in the range mentioned above are also possible (eg, about 1/5000 or more and less than 1/500 of the diffusivity of particles diffusing in water under the same conditions). Other ranges are possible. Measurements can be based on a time scale of about 1 second, about 0.5 seconds, about 2 seconds, about 5 seconds, or about 10 seconds.

In individual embodiments, the particles described herein are capable of diffusing into human cervical mucus at a diffusivity of about 1/500 or less of the diffusivity of the particles diffusing in water. In some cases, measurements can be based on a time scale of about 1 second, about 0.5 seconds, about 2 seconds, about 5 seconds, or about 10 seconds.

In certain embodiments, the present invention provides particles that move in mucus, such as human cervical mucus, at a particular absolute diffusion rate. For example, the particles described herein are at least about 1x10.^-4μm / sec, 2 × 10^-4μm / sec, 5 × 10^-4μm / sec, 1 × 10^-3μm / sec, 2 × 10^-3μm / sec, 5 × 10^-3μm / sec, 1 × 10^-2μm / sec, 2 × 10^-2μm / sec, 4 × 10^-2μm / sec, 5 × 10^-2μm / sec, 6 × 10^-2μm / sec, 8 × 10^-2μm / sec, 1 × 10^-1μm / sec, 2 × 10^-1μm / sec, 5 × 10^-1It can move at a diffusion rate of μm / sec, 1 μm / sec, or 2 μm / sec. In some cases, the particles are about 2 μm / sec or less, about 1 μm / sec or less, about 5 × 10^-1μm / sec or less, about 2 × 10^-1μm / sec or less, about 1 × 10^-1μm / sec or less, about 8 × 10^-2μm / sec or less, about 6 × 10^-2μm / sec or less, about 5 × 10^-2μm / sec or less, about 4 × 10^-2μm / sec or less, about 2 × 10^-2μm / sec or less, about 1 × 10^-2μm / sec or less, about 5 × 10^-3μm / sec or less, about 2 × 10^-3μm / sec or less, about 1 × 10^-3μm / sec or less, about 5 × 10^-4μm / sec or less, about 2 × 10^-4μm / sec or less, or about 1 × 10^-4It can move at a diffusion rate of μm / sec or less. Combinations of ranges mentioned above (eg about 2x10)^-4μm / sec or more and about 1 × 10^-1(Μm / sec or less) is also possible. Other ranges are possible. In some cases, the measurements are based on a time scale of about 1 second, about 0.5 seconds, about 2 seconds, about 5 seconds, or about 10 seconds.

It is understood that many of the mobilities described herein (eg, relative velocity, diffusivity) can be measured in human cervical mucus, but those mobilities can be measured in other types of mucus as well. sea bream.

In certain embodiments, the particles described herein contain a surface modification portion at a predetermined density. The surface modification portion may be, for example, a part of a surface modification agent exposed to a solvent containing particles. As an example, the PEG moiety can be a surface modification moiety of the surface modifier PEG-PPO-PEG. In some cases, the surface modification moiety and / or the surface modifier is nm.²At least about 0.001, at least about 0.002, at least about 0.005, at least about 0.01, at least about 0.02, at least about 0.05, at least about 0.1, at least about 0.2, at least about 0.5, at least about 1, at least about 1. 2, at least about 5, at least about 10, at least about 20, at least about 50, at least about 100, or more. In some cases, the surface modification moiety and / or the surface modifier is nm.²Per unit or number of molecules, about 100 or less, about 50 or less, about 20 or less, about 10 or less, about 5 or less, about 2 or less, about 1 or less, about 0.5 or less, about 0.2 or less, about 0.1 or less, about 0.05 Below, it exists at a density of about 0.02 or less, or about 0.01 or less. Combinations of ranges mentioned above (eg nm²At least about 0.01 and about 1 or less per unit or number of molecules) is also possible. Other ranges are possible. In some embodiments, the density value may be the average density when the surface modifier is present in equilibrium with other components in solution.

Those of skill in the art will know how to quantify or estimate the average density of surface modification (eg, each of which is incorporated herein by reference, SJ Budijono et al., Colloids and Surfaces). A: Physicochem. Eng. Aspects 360 (2020) 105-110, and Joshi, et al., Anal. Chim. Acta 104 (1979) see 153-160). For example, as described herein, the average density of surface modification can be measured using HPLC quantification and DLS analysis. Suspensions of particles for which surface density measurement is a challenge are first sized using DLS: dilute small amounts to the appropriate concentration (eg, about 100 μg / mL) and measure the z average diameter by particle size. It is regarded as a representative measurement value of. The remaining suspension is then split into two aliquots. Using HPLC, the first aliquot is assayed for the total concentration of core material and for the total concentration of surface modification. The second aliquot is assayed again for the concentration of free or unconstrained surface modification moieties using HPLC. Particles, and thus any constrained surface modification, are removed by ultracentrifugation in order to obtain only free or unconstrained surface modification from the second aliquot. By subtracting the concentration of the unconstrained surface-modified portion from the total concentration of the surface-modified portion, the concentration of the constrained surface-modified portion can be obtained. Since the total concentration of the core material was also determined from the first aliquot, the mass ratio between the core material and the surface-modified portion can be determined. The molecular weight of the surface-modified portion can be used to calculate the number of surface-modified portions with respect to the mass of the core material. In order to divert this number to the measurement of surface density, it is necessary to calculate the surface area per mass of the core material. The volume of the particles is approximated as that of a sphere with a diameter obtained from DLS that allows the calculation of the surface area per mass of the core material. With this method, the number of surface-modified portions per surface area can be determined. This method was used to measure the density values described herein.

Density also quantifies or estimates the surface area of the core material by methods such as electron microscopy, light scattering, or measurement of surface interactions, followed by surface modification adhered by methods such as liquid chromatography or mass spectroscopy. It can be measured by quantifying the agent. (See, for example, Wang et al., Angew Chem Int Ed Engl, 2008, 47 (50), 9726-9, incorporated herein by reference).

In certain embodiments, the particles described herein comprise a surface modification moiety and / or a surface modifier that affects the zeta potential of the particle. The zeta potentials of the coated particles are, for example, at least about -100 mV, at least about -75 mV, at least about -50 mV, at least about -40 mV, at least about -30 mV, at least about -20 mV, at least about -10 mV, at least about -5 mV. , At least about 5 mV, at least about 10 mV, at least about 20 mV, at least about 30 mV, at least about 40 mV, at least about 50 mV, at least about 75 mV, or at least about 100 mV. Combinations in the range mentioned above (eg, zeta potentials of at least about -50 mV and less than about 50 mV) are also possible. Other ranges are possible.

The coated particles described herein can have any suitable shape and / or size. In some embodiments, the coated particles have a shape substantially similar to the shape of the core. In some cases, the coated particles described herein may be nanoparticles, i.e., the particles have characteristic dimensions of about 1 μm or less, and the characteristic dimensions of the particles are the particles. The diameter of a perfect sphere with the same volume. In other embodiments, larger sizes are possible (eg, about 1-10 μm). Multiple particles can also be characterized by average size in some embodiments (eg, average maximum cross-sectional dimension or average minimum cross-sectional dimension for multiple particles). Multiple particles are, for example, about 10 μm or less, about 5 μm or less, about 1 μm or less, about 800 nm or less, about 700 nm or less, about 500 nm or less, 400 nm or less, 300 nm or less, about 200 nm or less, about 100 nm or less, about 75 nm or less, about. It can have an average size of 50 nm or less, about 40 nm or less, about 35 nm or less, about 30 nm or less, about 25 nm or less, about 20 nm or less, about 15 nm or less, or about 5 nm or less. In some cases, the particles may be, for example, at least about 5 nm, at least about 20 nm, at least about 50 nm, at least about 100 nm, at least about 200 nm, at least about 300 nm, at least about 400 nm, at least about 500 nm, at least about 1 μm, or It can have an average size of at least about 5 μm. Combinations in the range mentioned above (eg, average size of at least about 50 nm and about 500 n or less) are also possible. Other ranges are possible. In some embodiments, the size of the core formed by the method described herein has a Gaussian distribution.

Pharmaceuticals In some embodiments, the coated particles comprise at least one pharmaceutical. The medicinal product can be present in the core of the particle and / or in the coating of the particle (eg, dispersed throughout the core and / or coating). In some cases, the medicinal product can be placed on the surface of the particles (eg, on the outer surface of the coating, on the inner surface of the coating, on the surface of the core). The medicinal product can be encapsulated within the particle and / or placed in part of the particle using commonly known techniques (eg, coating, adsorption, covalent bond, or other method). In some cases, the drug can be present in the core of the particle before or during coating the particle. In some cases, the drug is present in forming the core of the particles, as described herein.

Non-limiting examples of pharmaceuticals include contrasting agents, diagnostic agents, therapeutic agents, drugs with detectable labels, nucleic acids, nucleic acid analogs, small molecules, peptide mimetics, proteins, peptides, lipids, vaccines, etc. Includes viral vectors, viruses, and surfactants.

In some embodiments, the pharmaceuticals included in the particles described herein have therapeutic, diagnostic, or contrast-enhancing effects in the mucosal tissue to be targeted. Non-limiting examples of mucosal tissue include the oral cavity (including, for example, the oral and esophageal membrane and tongue surface), eyes, gastrointestinal tract (including, for example, stomach, small intestine, large intestine, colon, rectum), nose, respiratory organs (eg, including stomach, small intestine, large intestine, colon, rectum). For example, tissues of the nose, pharynx, trachea and bronchi), and reproductive organs (eg, vaginal, cervical and esophageal membranes).

Any suitable number of medicines can be present in the particles described herein. For example, at least one, at least two, at least three, at least four, at least five, or more, but generally less than ten, are present in the particles described herein. Can be done.

Many drugs that are mucous and sticky are known in the art and can be used as pharmaceuticals in the particles described herein (eg, Khanvilkar K, Donovan MD, Flanagen DR, Drug transfer through mucus). , Advanced Drug Delivery Reviews 48 (2001) 173-193; Bhat PG, Flanagan DR, Donovan MD. Drug diffusion through cystic fibrotic mucus: steady-state permeation, rheologic properties, and glycoprotein morphology, J Pharm Sci, 1996 Jun; 85 (6): 624-30). Further non-limiting examples of pharmaceuticals include contrast and diagnostic agents (radio impermeable agents, labeled antibodies, labeled nucleic acid probes, dyes such as colored or fluorescent dyes, etc.), and adjuvants (radiosensitizers, etc.). Transfection enhancers, mobilizing agents and migrating attractants, peptides that regulate cell adhesion and / or cell mobility, cell penetrants, vaccine enhancers, multidrug resistance inhibitors and / or outflow pump inhibitors , Etc.) are included.

Further non-limiting examples of pharmaceuticals include alloxypurine, auranofin, azapropazone, benolylate, diflunisal, etodolac, fenbufen, fenoprofen calcium (calcim), flurubiprofen, frosemide, ibuprofen, indomethacin, ketoprofen, rotepredonol etabonate, Meclophenamic acid, mefenamic acid, nabmeton, naproxen, oxyphenbutazone, phenylbutazone, pyroxicum, sulindac, alvendazole, bephenium hydroxynaphthoate, cambendazole, dichlorophene, ibuprofen, mebendazole, oxamnikin, oxfendazole, oxendazole embonate. , Praziquantel, pyrantel embonate, thiabendazole, amyodaron HCl, disopyramide, flecanide acetate, kinidine sulfate. Antibacterial agent:

Applications and Pharmaceutical Compositions The particles described herein can be employed in any suitable application. In some cases, the particles are a mucous or mucosal surface of a pharmaceutical composition (eg, as described herein), eg, a drug (eg, drug, therapeutic agent, diagnostic agent, contrast agent). Part of a pharmaceutical composition used through or to deliver to them. The pharmaceutical composition can include at least one of the particles described herein, and one or more pharmaceutically acceptable excipients or carriers. The composition can be used in treating, preventing, and / or diagnosing the condition of the subject, the method comprising administering the pharmaceutical composition to the subject. The subject or patient to be treated by the articles and methods described herein can mean human or non-human animals such as primates, mammals, and vertebrates.

Methods associated with treating a subject may be predisposed to the disease, disorder and / or condition, but prevent the development of the disease, disorder or condition in a subject who has not been diagnosed with them; It can include suppressing a disease, disorder or condition, eg, slowing its progression; and alleviating the disease, disorder or condition, eg, resulting in regression of the disease, disorder and / or condition. Treating a disease or condition improves at least one symptom of an individual disease or condition, even if it does not affect the underlying pathophysiology (eg, such agents cause pain). Includes (such as treating a subject's pain by administering an analgesic, if not treated).

In some embodiments, the pharmaceutical compositions described herein are delivered to a mucosal surface in a subject and pass through a mucosal barrier in the subject (eg, mucus), eg, due to reduced mucous stickiness. And / or can exhibit a long retention of particles and / or an increase in uniform distribution on the mucosal surface. Non-limiting examples of mucosal tissue include the oral cavity (including, for example, the buccal and esophageal membranes, and the surface of the tonsils), the eye, the gastrointestinal tract (including, for example, the stomach, small intestine, large intestine, colon, rectum), nasal cavity, etc. Includes respiratory tract (eg, including nasal cavity, pharynx, trachea and bronchial membranes), genital organs (eg, including vaginal, cervical and esophageal membranes).

Pharmaceutical compositions described herein and for use in accordance with the articles and methods described herein can include pharmaceutically acceptable excipients or carriers. As a pharmaceutically acceptable excipient or pharmaceutically acceptable carrier, any suitable type of non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or pharmaceutical adjunct. The agent can be mentioned. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars (such as lactose, glucose and sucrose), starches (such as corn starch and potato starch), cellulose and its derivatives (sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate). Excipients such as powdered tragacanto rubber, malt, gelatin, talc, cacao butter and suppository wax, oils (peanut oil, cottonseed oil, benibana oil, sesame oil, olive oil, corn oil and soybean oil, etc.), glycol (propylene glycol). , Esters (such as ethyl oleate and ethyl laurate), agar, surfactants (such as Tween80), buffers (such as magnesium hydroxide and aluminum hydroxide), alginic acid, pyrogen-free, isotonic physiological saline, etc. Ringer solution, ethyl alcohol, phosphate buffer solution, other non-toxic and compatible lubricants (such as sodium lauryl sulfate and magnesium stearate), colorants, release agents, coatings, sweeteners, flavoring agents. And fragrances, preservatives and antioxidants may also be present in the composition at the discretion of the pharmaceutical manufacturer. As will be appreciated by those of skill in the art, excipients can be selected based on the route of administration, the drug delivered, the time course of drug delivery, etc., as described below.

The pharmaceutical composition comprising the particles described herein can be administered to a subject via any route known in the art. These routes are, but not limited to, oral, sublingual, nasal, intradermal, subcutaneous, intramuscular, transrectal, transvaginal, intravenous, intraarterial, intracisional, intraperitoneal, intravitreal, and periocular. , Topical (as a powder, cream, ointment, or drip), buccal, and inhaled administration. In some embodiments, the compositions described herein can be administered parenterally, such as injectables (intravenous, intramuscular or subcutaneous), infusion formulations, or suppositories. As will be appreciated by those of skill in the art, the route of administration and effective dosage to achieve the desired biological effect will be the drug to be administered, the target organ, the formulation to be administered, the time course of administration, the treatment. It can be determined according to the disease, intended use, etc.

As an example, particles can be included in a pharmaceutical composition to be formulated as a nasal spray such that the pharmaceutical composition is delivered across the nasal mucus layer. As another example, the particles can be included in the pharmaceutical composition to be formulated as an inhalant such that the pharmaceutical composition is delivered across the pulmonary mucus layer. As another example, if the composition is to be administered orally, the composition can be formulated as a tablet, capsule, granule, powder, or syrup. Similarly, the particles can be included in a pharmaceutical composition to be delivered via the eye, gastrointestinal tract, nasal cavity, respiratory tract, rectum, urethra and / or vaginal tissue.

For application by the ocular mucomembrane pathway, the subject composition can be formulated as an eye drop or an ophthalmic ointment. These formulations can be prepared by conventional means and, if desired, the subject composition can be buffered or pH regulated, tonicity regulators, viscosity regulators, suspension stabilizers, preservatives, and others. It can be mixed with any conventional additive, such as a pharmaceutical excipient. Further, in certain embodiments, the subject compositions described herein can be freeze-dried or subjected to another suitable drying technique such as spray drying.

In some embodiments, the particles described herein that can be administered in the form of an inhalant or aerosol formulation are one of the adjuvants, diagnostic agents, contrast agents, or therapeutic agents useful in inhalation therapy. Or includes multiple medications. The particle size of the particulate drug should allow inhalation of virtually all drugs into the lungs by administration of the aerosol formulation, eg, about 20 μm or less, eg, about 1 to about 10 μm, For example, it may be about 1 to about 5 μm, but other ranges are possible. The particle size of the drug can be reduced by conventional means, for example by grinding or micronization. Alternatively, the particulate agent can be administered to the lungs by atomizing the suspension. The final aerosol formulation is by weight / weight% by weight, eg 0.005 to 90%, 0.005 to 50%, 0.005 to 10%, about 0.005 to 5%, or 0.01 to 1.0% of the total weight of the formulation. Can be included. Other ranges are possible.

The formulations described herein should be free of, but not essential, components that may induce the decomposition of ozone in the stratosphere. In particular, in some embodiments, CCl₃F, CCl₂F₂, And CF₃CCl₃Injections that do not contain or are essentially composed of chlorofluorocarbons, such as, are selected.

Aerosols can include propellants. The propellant may contain an adjunct having a higher polarity and / or a higher boiling point than the propellant. Polar adjuvants that can be used include (eg, C)._2-6) Aliphatic alcohols and polyols, such as ethanol, isopropanol and propylene glycol, preferably ethanol. In general, only small amounts (eg, 0.05-3.0% by weight / weight%) of polar aids may be needed to improve the stability of the dispersion, and when used in excess of 5% by weight / weight, It may tend to dissolve the drug. The formulations according to the embodiments described herein can include less than about 1% by weight / weight, eg, about 0.1% by weight / weight of polar adjuvant. However, the formulations described herein may be substantially free of polar aids, especially ethanol. Suitable volatile adjuvants include saturated hydrocarbons such as propane, n-butane, isobutane, pentane and isopentane, and alkyl ethers such as dimethyl ether. Generally, volatile aids up to 50% by weight / weight of propellant, eg 1-30% by weight / weight% volatile saturated C₁-C₆It can contain hydrocarbons. Optionally, the aerosol formulation according to the invention can further comprise one or more surfactants. Surfactants may be physiologically acceptable for administration by inhalation. Examples of the surfactant included in this category include L-α-phosphatidylcholine (PC), 1,2-dipalmitoylphosphatidylcholine (DPPC), oleic acid, sorbitan trioleate, sorbitan monooleate, and sorbitan monolaurate. Polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, natural lecithin, oleyl polyoxyethylene ether, stearyl polyoxyethylene ether, lauryl polyoxyethylene ether, block copolymer of oxyethylene and oxypropylene, synthetic lecithin, diolein Diethylene glycol acid, tetrahydrofurfuryl oleate, ethyl oleate, isopropyl myristate, glyceryl monooleate, glyceryl monostearate, glyceryl monolithinol, cetyl alcohol, stearyl alcohol, polyethylene glycol 400, cetylpyridinium chloride, benzalconium chloride Includes, oleic acid, glyceryl monolaurate, corn oil, cottonseed oil, and sunflower seed oil.

The formulations described herein can be prepared by dispersing the particles in a selected propellant and / or propellant in a suitable container, eg, with the help of sonication. .. The particles can be suspended in a propellant and filled in a suitable container. The valve of the container is then mounted in place, pressurized with the propellant and introduced by filling through the valve in a conventional manner. Thus, the particles are suspended or dissolved in a liquefied propellant and sealed in a container equipped with a metering valve and suitable for the actuating device. Such fixed dose inhalers are well known in the art. The weighing valve can weigh and supply 10 to 500 μL, preferably 25 to 150 μL. In certain embodiments, dispersion can be achieved using a dry powder inhaler (eg, a rotary inhaler) for the particles (remaining as a dry powder). In another embodiment, the nanospheres can be suspended in an aqueous fluid and atomized into microdroplets that will be aerosolized in the lungs.

Sonic nebulizers can also be used to minimize exposure of the agent to shears that can result in particle degradation. Aqueous aerosols are usually prepared by formulating an aqueous solution or suspension of particles with a pharmaceutically acceptable conventional carrier and stabilizer / surface modifier. Carriers and stabilizers / surface modifiers will vary depending on the requirements of the individual composition, but are typically such as nonionic surfactants (Tween, Pluronic®, or polyethylene glycol), serum albumins. Harmless proteins, amino acids such as sorbitan esters, oleic acid, lecithin, glycine, buffers, salts, sugars, or sugar alcohols. Aerosols are generally prepared from isotonic solutions.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient (ie, microparticles, nanoparticles, liposomes, micelles, polynucleotide / lipid complexes), the liquid dosage form is an inert diluent commonly used in the art, such as water or Other solvents; solubilizers and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, etc.) It can include peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohols, esters of polyethylene glycol of sorbitan with fatty acids, and combinations thereof. In addition to the Inactive Diluent, the oral composition can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, and aromatics.

Injectable formulations, such as injectable sterile aqueous or oily suspensions, can be formulated by known techniques using appropriate dispersion or wetting agents and suspending agents. Injectable sterile formulations may also be parenterally acceptable non-toxic diluents or solutions in solvents Injectable solutions, suspensions, or emulsions, such as solutions in 1,3-butanediol. Acceptable media and solvents that can be adopted include water, Ringer's solution, United States Pharmacopeia's solution, and isotonic sodium chloride solution, among others. In addition, sterile non-volatile oils are commonly employed as the solvent or suspension medium. For this purpose, any non-volatile non-volatile oil, such as synthetic mono or diglycerides, can be employed. In addition, fatty acids such as oleic acid are used in the formulation of injectable materials. In certain embodiments, the particles are suspended in a carrier fluid containing 1 (weight / volume)% sodium carboxymethyl cellulose and 0.1 (volume / volume)% Tween80.

Sterilizers in the form of sterile solid compositions that allow the injectable formulation to be dissolved or dispersed, for example, by filtration through a bacterial retention filter or in sterile water or other injectable sterile medium prior to use. Can be sterilized by incorporating.

Compositions for transrectal or vaginal administration are solid at external temperature, but liquid at body temperature and therefore melt and release particles in the rectal or vaginal cavity, cocoa butter, polyethylene glycol, or. A suppository may be prepared by mixing the particles with a suitable non-irritating excipient such as suppository wax or carrier.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the particles are composed of at least one inert and pharmaceutically acceptable excipient or carrier, such as sodium citrate or dipotassium phosphate, and / or a) starch, lactose, and the like. Fillers or bulking agents such as sucrose, glucose, mannitol, and silicic acid, b) binders such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic, c) moisturizers such as glycerol, etc. d) Disintegrants such as agar, calcium carbonate, horse bell Potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) Nojima dissolution retarders such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and these. It is mixed with a lubricant such as a mixture of. For capsules, tablets, and pills, the dosage form can also include a buffer.

Similar types of solid compositions can also be employed as fillers in soft and hard filled gelatin capsules using excipients such as lactose or lactose, and high molecular weight polyethylene glycol.

Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmaceutical formulations. They can optionally contain an opacity agent and can also have a composition that releases only the active ingredient in a particular portion of the intestinal tract, or preferentially delays them. Examples of embedding compositions that can be used are polymeric substances and waxes.

Similar types of solid compositions can also be employed as fillers in soft and hard filled gelatin capsules using excipients such as lactose or lactose, and high molecular weight polyethylene glycol.

Dosage forms for topical or transdermal administration of the pharmaceutical composition of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. Can be mentioned. The particles are mixed under sterile conditions with a pharmaceutically acceptable carrier and, if necessary, any required preservative or buffer. Ophthalmic formulations, ear drops, and eye drops are also considered within the scope of the present invention.

Ointments, pastes, creams, and gels, in addition to the particles described herein, include animal and vegetable fats, oils, waxes, paraffins, starches, tragacant rubbers, cellulose derivatives, polyethylene glycols, silicones, etc. Excipients such as bentonite, silicic acid, starch, and zinc oxide, or mixtures thereof, can be included.

Powders and sprays include, in addition to the particles described herein, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these substances. Can be done. The atomizer can further include conventional propellants such as chlorofluorohydrocarbons.

Transdermal patches have the additional advantage of providing controlled delivery of the compound to the body. Such dosage forms can be prepared by dissolving or dispersing microparticles or nanoparticles in a suitable medium. Absorption enhancers can be used to increase the flow of compounds across the skin. The velocity can be adjusted by preparing a velocity adjusting membrane or by dispersing the particles in a polymer matrix or gel.

The particles described herein, including the drug, are administered to the subject to be delivered in an amount sufficient to deliver to the subject a therapeutically effective incorporated dose of the drug as part of a diagnostic, prophylactic or therapeutic procedure. can do. In general, the effective amount of a drug or ingredient refers to the amount required to elicit the desired biological response. The desired concentration of the drug in the particles is, but is not limited to, the rate of absorption, inactivation, and excretion of the drug, as well as the rate of delivery of the compound from the composition of interest, the desired biological ultimate goal, the drug to be delivered, It depends on many factors such as target tissue. It should be noted that the dose value also depends on the severity of the condition to be alleviated. In addition, it should be understood that the specific dosing regimen should be adjusted over time for any particular subject, at the discretion of the individual's needs and the professional judgment of the person who controls or supervises the administration of the composition. .. Administration is typically determined using techniques known to those of skill in the art.

The pharmaceutical product is in the composition and / or the formulation in any suitable amount, eg, at least about 0.01% by weight, at least about 0.1% by weight, at least about 1% by weight, at least about 5% by weight of the composition and / or the formulation. It can be present in%, at least about 10% by weight, and at least about 20% by weight. In some cases, the medicinal product in the composition and / or formulation is about 30% by weight or less, about 20% by weight or less, about 10% by weight or less, about 5% by weight or less, about 2% by weight or less, or about. Can be present in less than 1% by weight. Combinations in the range mentioned above are also possible (eg, present in an amount of at least about 0.1% by weight and about 10% by weight or less). Other ranges are possible. In certain embodiments, the pharmaceutical product is present in about 0.1-2% by weight of the composition and / or the formulation. In certain embodiments, the pharmaceutical product is present in about 2-20% by weight of the composition and / or the formulation. In certain embodiments, the pharmaceutical product is present in about 0.2% by weight of the composition and / or the formulation. In certain embodiments, the pharmaceutical product is present in about 0.4% by weight of the composition and / or the formulation. In certain embodiments, the pharmaceutical product is present in about 1% by weight of the composition and / or the formulation. In certain embodiments, the pharmaceutical product is present in about 2% by weight of the composition and / or the formulation. In certain embodiments, the pharmaceutical product is present in about 5% by weight of the composition and / or the formulation. In certain embodiments, the pharmaceutical product is present in about 10% by weight of the composition and / or the formulation.

Any of the concentrations and / or amounts of the drug to be administered to the subject can be readily determined by one of ordinary skill in the art. Also known methods are available for assaying concentrations in local tissues, diffusion rates from particles, and local blood flow before and after administration of the therapeutic formulation.

The compositions and / or formulations described herein can have any suitable volume Osmar concentration. In some embodiments, the compositions and / or formulations described herein are at least about 0 mOsm / L, at least about 5 mOsm / L, at least about 25 mOsm / L, at least about 50 mOsm / L, and at least about 75 mOsm /. It can have a capacity Osmar concentration of L, at least about 100 mOsm / L, at least about 150 mOsm / L, at least about 200 mOsm / L, at least about 250 mOsm / L, or at least about 310 mOsm / L. In certain embodiments, the compositions and / or formulations described herein are about 310 mOsm / L or less, about 250 mOsm / L or less, about 200 mOsm / L or less, about 150 mOsm / L or less, about 100 mOsm / L or less, It can have a volume Osmar concentration of about 75 mOsm / L or less, about 50 mOsm / L or less, about 25 mOsm / L or less, or about 5 mOsm / L or less. Combinations in the range mentioned above (eg, volume Osmar concentrations of at least about 0 mOsm / L and about 50 mOsm / L or less) are also possible. Other ranges are possible. The volume Osmar concentration of the composition and / or the formulation can be changed, for example, by changing the concentration of the salt present in the solvent of the composition and / or the formulation.

In a set of embodiments, the composition and / or the formulation comprises one or more chelating agents. The chelating agent used in the present invention refers to a chemical compound capable of reacting with a metal ion to form a complex via one or more bonds. One or more bonds are typically ionic or coordinate bonds. The chelating agent may be an inorganic or organic compound. A metal ion capable of catalyzing a particular chemical reaction (eg, an oxidation reaction) may lose its catalytic activity when the metal ion is bound to a chelating agent to form a complex. Therefore, the chelating agent may exhibit shelf life when it is bound to a metal ion. Any suitable chelating agent with storage stability can be used, such as phosphonic acid, aminocarboxylic acid, hydroxycarboxylic acid, polyamines, aminoalcohols, and polymeric chelating agents. Specific examples of the chelating agent include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), diethylenetriaminepentaacetic acid (DTPA), N-hydroxyethylethylenediaminetriacetic acid (HEDTA), tetraborate, Includes triethylaminediamine, as well as salts and derivatives thereof. In certain embodiments, the chelating agent is EDTA. In certain embodiments, the chelating agent is a salt of EDTA. In certain embodiments, the chelating agent is EDTA disodium.

The chelating agent can be present at an appropriate concentration in the composition and / or the formulation containing the coated particles described herein. In certain embodiments, the concentration of chelating agent is about 0.0003% by weight or more, about 0.001% by weight or more, about 0.003% by weight or more, about 0.01% by weight or more, about 0.03% by weight or more, about 0.05% by weight or more, about. 0.1% by weight or more, about 0.3% by weight or more, about 1% by weight or more, or about 3% by weight or more. In certain embodiments, the concentrations of the chelating agent are about 3% by weight or less, about 1% by weight or less, about 0.3% by weight or less, about 0.1% by weight or less, about 0.05% by weight or less, about 0.03% by weight or less, about 0.03% by weight or less. 0.01% by weight or less, about 0.003% by weight or less, about 0.001% by weight or less, or about 0.0003% by weight or less. Combinations in the range mentioned above (eg, concentrations of about 0.01% by weight or more and about 0.3% by weight or less) are also possible. Other ranges are possible. In certain embodiments, the concentration of chelating agent is from about 0.001 to 0.1% by weight. In certain embodiments, the concentration of chelating agent is about 0.005% by weight. In certain embodiments, the concentration of chelating agent is about 0.01% by weight. In certain embodiments, the concentration of chelating agent is about 0.05% by weight. In certain embodiments, the concentration of chelating agent is about 0.1% by weight.

In some embodiments, the chelating agent is present in the composition and / or the formulation to the extent mentioned above, during the forming and / or diluting steps described herein. be able to. In certain embodiments, the chelating agent can be present in the final product, in the composition and / or the formulation, to the extent mentioned above, one or more.

In some embodiments, antimicrobial agents can be included in compositions and / or formulations comprising the coated particles described herein. The antimicrobial agent used in the present invention is used to inhibit, prevent, or protect from microorganisms such as bacteria, microorganisms, fungi, viruses, spores, yeasts, molds, and other microorganisms commonly associated with infectious diseases. Refers to an effective bioactive agent. Examples of antimicrobial agents include cephalosporin, clindamycin, chlorampheanicol, carbapenem, minocycline, riffampin, penicillin, monbactam, quinolone, tetracycline, macrolides, sulfa antibiotics, trimetoprim, fusidic acid, Aminoglycoside, amphotelicin B, azole, flucitosine, sirofungin, bactericidal nitrofuran compound, nanoparticles of silver alloy containing metallic silver or about 2.5% by weight copper, silver citrate, silver acetate, silver benzoate, bismuspyrthione, zincpyrythion , Zinc percarbonate, zinc perborate, bismuth salt, parabens (eg, methyl, ethyl, propyl, butyl and octyl esters of benzoic acid), citric acid, benzalconium chloride (BAC), rifamycin, and sodium percarbonate. Is included.

The antimicrobial agent can be present at the appropriate concentration in the composition and / or the formulation comprising the coated particles described herein. In certain embodiments, the concentrations of the antimicrobial agent are about 0.0003% by weight or more, about 0.001% by weight or more, about 0.003% by weight or more, about 0.01% by weight or more, about 0.03% by weight or more, about 0.1% by weight or more, about 0.1% by weight. It may be 0.3% by weight or more, about 1% by weight or more, or about 3% by weight or more. In certain embodiments, the concentrations of the antimicrobial agent are about 3% by weight or less, about 1% by weight or less, about 0.3% by weight or less, about 0.1% by weight or less, about 0.03% by weight or less, about 0.01% by weight or less, about. It may be 0.003% by weight or less, about 0.001% by weight or less, or about 0.0003% by weight or less. Combinations in the range mentioned above (eg, concentrations of about 0.001% by weight or more and about 0.1% by weight or less) are also possible. Other ranges are possible. In certain embodiments, the concentration of antimicrobial agent is from about 0.001 to 0.05% by weight. In certain embodiments, the concentration of antimicrobial agent is about 0.002% by weight. In certain embodiments, the concentration of antimicrobial agent is about 0.005% by weight. In certain embodiments, the concentration of antimicrobial agent is about 0.01% by weight. In certain embodiments, the concentration of antimicrobial agent is about 0.02% by weight. In certain embodiments, the concentration of antimicrobial agent is about 0.05% by weight.

In some embodiments, the antimicrobial agent is present in the composition and / or the formulation to the extent mentioned above, during the forming and / or diluting steps described herein. be able to. In certain embodiments, the antimicrobial agent can be present in the final product, in the composition and / or in the formulation, to the extent mentioned above, one or more.

In some embodiments, isotonic agents can be included in compositions and / or formulations comprising the coated particles described herein. The tonicity agent used in the present invention refers to a compound or substance that can be used to adjust the composition of a pharmaceutical product to a desired volume Osmar concentration range. In certain embodiments, the desired volume Osmar concentration range is an isotonic range compatible with blood. In certain embodiments, the desired volume Osmar concentration range is hypotonic. In certain embodiments, the desired volume Osmar concentration range is hypertonic. Examples of isotonic agents include glycerin, lactose, mannitol, dextrose, sodium chloride, sodium sulphate, sorbitol, saline-sodium citrate (SSC) and the like. In certain embodiments, one or a combination of isotonic agents can be used. In certain embodiments, the tonicity agent is glycerin. In certain embodiments, the tonicity agent is sodium chloride.

The tonicity agent (as described herein) can be present at appropriate concentrations in compositions and / or formulations containing the coated particles described herein. In certain embodiments, the isotonic agent concentration is about 0.003% by weight or more, about 0.01% by weight or more, about 0.03% by weight or more, about 0.1% by weight or more, about 0.3% by weight or more, about 1% by weight or more, About 3% by weight or more, about 10% by weight or more, about 20% by weight or more, or about 30% by weight or more. In certain embodiments, the concentrations of the tonicity agent are about 30% by weight or less, about 10% by weight or less, about 3% by weight or less, about 1% by weight or less, about 0.3% by weight or less, about 0.1% by weight or less, It is about 0.03% by weight or less, about 0.01% by weight or less, or about 0.003% by weight or less. Combinations in the range mentioned above (eg, concentrations of about 0.1% by weight or more and about 10% by weight or less) are also possible. Other ranges are possible. In certain embodiments, the concentration of isotonic agent is about 0.1-1%. In certain embodiments, the concentration of isotonic agent is about 0.5-3%. In certain embodiments, the isotonic agent concentration is about 0.25% by weight. In certain embodiments, the isotonic agent concentration is about 0.45% by weight. In certain embodiments, the isotonic agent concentration is about 0.9% by weight. In certain embodiments, the isotonic agent concentration is about 1.2% by weight. In certain embodiments, the isotonic agent concentration is about 2.4% by weight. In certain embodiments, the isotonic agent concentration is about 5% by weight.

In some embodiments, the tonicity agent is present in the composition and / or formulation to the extent mentioned above, during the forming and / or diluting steps described herein. be able to. In certain embodiments, the tonicity agent can be present in the final product, to the extent mentioned above, in the composition and / or the formulation.

Polydispersity is a measure of particle size non-uniformity in a pharmaceutical product. The non-uniformity of the particle size may be due to the difference in individual particle size and / or the presence of agglomeration in the formulation. A pharmaceutical product containing particles is considered to be substantially uniform or "monodisperse" if the particles have essentially the same size, shape, and / or mass. Formulations containing particles of various sizes, shapes, and / or masses are considered non-uniform or "polydisperse".

In some embodiments, the polydispersity of the composition and / or the formulation keeps the added ionic strength of the composition and / or the formulation relatively constant in the presence of the added ionic strength. It is relatively constant when it is added or increased (eg, during the forming and / or dilution steps). In certain embodiments, when the ionic strength is increased to at least 50%, the polydispersity is about 200% or less, about 150% or less, about 100% or less, about 75% or less, about 50% or less, about 30% or less. , About 20% or less, about 10% or less, about 3% or less, or up to about 1% or less. In certain embodiments, increasing the ionic strength to at least 50% increases the polydispersity to about 1% or more, about 3% or more, about 10% or more, about 30% or more, or about 100% or more do. Combinations in the range mentioned above (eg, 50% or less and 1% or more polydispersity increase) are also possible. Other ranges are possible.

The ionic strength of the formulations described herein can be adjusted via various means such as adding one or more ionic isotonic agents (eg, salts such as NaCl) to the formulations. .. In certain embodiments, the ionic strengths of the formulations described herein are about 0.0005 M and above, about 0.001 M and above, about 0.003 M and above, about 0.01 M and above, about 0.03 M and above, about 0.1 M and above, about 0.3. M or more, about 1M or more, about 3M or more, or about 10M or more. In certain embodiments, the ionic strengths of the formulations described herein are about 10 M or less, about 3 M or less, about 1 M or less, about 0.3 M or less, about 0.1 M or less, about 0.03 M or less, about 0.01 M or less, It is about 0.003M or less, about 0.001M or less, or about 0.0005M or less. Combinations in the range mentioned above (eg, ionic strength of about 0.01 M or more and about 1 M or less) are also possible. Other ranges are possible. In certain embodiments, the ionic strength of the formulations described herein is about 0.1 M. In certain embodiments, the ionic strength of the formulations described herein is about 0.15 M. In certain embodiments, the ionic strength of the formulations described herein is about 0.3 M.

The polydispersity of the formulations described herein can be measured by the polydispersity index (PDI). The PDI is used to describe the width of the particle size distribution and is often calculated from a cumulant analysis of dynamic light scattering (DLS) measured using the automatic intensity correction feature. Calculations for these parameters are specified in Standards ISO13321: 1996E and ISO22412: 2008. PDI is dimensionless and, when measured by DLS, values below 0.05 are calibrated to indicate highly monodisperse samples, and values above 0.7 are graduated to show a very wide particle size distribution. In certain embodiments, the PDIs of the formulations and / or compositions described herein are about 1 or less, about 0.9 or less, about 0.8 or less, about 0.7 or less, about 0.6 or less, about 0.5 or less, about 0.4 or less, It is about 0.3 or less, about 0.2 or less, about 0.15 or less, about 0.1 or less, about 0.05 or less, about 0.01 or less, or about 0.005 or less. In certain embodiments, the PDIs of the formulations and / or compositions described herein are about 0.005 and above, about 0.01 and above, about 0.05 and above, about 0.1 and above, about 0.15 and above, about 0.2 and above, about 0.3 and above, About 0.4 or more, about 0.5 or more, about 0.6 or more, about 0.7 or more, about 0.8 or more, about 0.9 or more, or about 1 or more. Combinations in the range mentioned above (eg, PDIs greater than or equal to about 0.1 and less than or equal to about 0.5) are also possible. Other ranges are possible. In certain embodiments, the PDI of the formulation is about 0.1. In a particular embodiment, the PDI of the formulation is about 0.15. In certain embodiments, the PDI of the formulation is about 0.2.

In certain embodiments, the compositions and / or formulations described herein can be highly dispersible and do not tend to form aggregates. Even when the particles form aggregates, the aggregates can be easily disassembled into individual particles without vigorous stirring of the composition and / or the formulation.

The compositions and / or formulations described herein can have any suitable pH value. The term "pH" refers to pH measured at outside temperature (eg, about 20 ° C, about 23 ° C, or about 25 ° C), unless otherwise noted. The composition and / or the formulation has, for example, an acidic pH, a neutral pH, or a basic pH, depending on the location in the body to which the composition and / or the formulation is to be delivered, for example. there's a possibility that. In certain embodiments, the composition and / or the pharmaceutical product has a physiological pH. In certain embodiments, the pH values of the composition and / or the formulation are at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 6.2, at least about 6.4, at least. About 6.6, at least about 6.8, at least about 7, at least about 7.2, at least about 7.4, at least about 7.6, at least about 7.8, at least about 8, at least about 8.2, at least about 8.4, at least about 8.6, at least about 8.8, at least about 9. , At least about 10, at least about 11, or at least about 12. In certain embodiments, the pH values of the compositions and / or formulations are about 12 or less, about 11 or less, about 10 or less, about 9 or less, about 8.8 or less, about 8.6 or less, about 8.4 or less, about 8.2 or less, about. 8 or less, about 7.8 or less, about 7.6 or less, about 7.4 or less, about 7.2 or less, about 7 or less, about 6.8 or less, about 6.6 or less, about 6.4 or less, about 6.2 or less, about 6 or less, about 5 or less, about 4 or less , About 3 or less, about 2 or less, or about 1 or less. Combinations in the range mentioned above (eg, pH values of at least about 5 and about 8.2 or less) are also possible. Other ranges are possible. In certain embodiments, the pH values of the compositions and / or formulations described herein are at least about 5 and not more than about 8.

In one set of embodiments, a composition comprising a plurality of coated particles is provided. The coated particles contain a solid drug or salt thereof or have a core formed from them, the drug or salt thereof is at about 1 mg / mL or less at 25 ° C anywhere during the pH range. It has a water solubility of (eg, about 0.1 mg / mL or less at 25 ° C) and at least about 80% by weight of the core particles (eg, at least about 90% by weight, at least about 95% by weight, at least about 99% by weight). To configure. The coated particles also comprise a coating comprising a surface modifier surrounding the core particles, the surface modifier comprising a triblock copolymer comprising a hydrophilic block-hydrophobic block-hydrophilic block arrangement, the hydrophobic block. Have a molecular weight of at least about 3 kDa, the hydrophilic block constitutes at least about 30% by weight of the triblock, and the hydrophilic block is or contains poly (ethylene oxide) having a molecular weight of at least about 2 kDa and is hydrophobic. The sex block associates with the surface of the core particle (eg, by adsorption), the hydrophilic block is present on the surface of the coated particle and makes the coated particle hydrophilic, and the surface modifier is on the surface of the core particle. At least about 0.001 molecule / nm²(For example, at least about 0.01 molecule / nm²) Exists. The coated particles have a relative velocity greater than 0.5 in mucus. The coated particles can have an average size of at least about 20 nm and about 1 μm or less (eg, about 500 nm or less). The coating thickness may be, for example, about 50 nm or less (eg, about 30 nm or less, about 10 nm or less). The composition may be a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, additives, and / or diluents. In some embodiments, the plurality of coated particles are one or more free surface modifiers in the composition (eg, Pluronic® P123, Pluronic® P103, Pluronic®). It exists in the form of a solution (eg, an aqueous solution) with P105, Pluronic® F127, Pluronic® F108, and a poloxamer selected from combinations thereof. The free surface modifier in solution and the surface modifier on the particle surface may be the same surface modifier and may be present in equilibrium with each other in the composition. The total amount of the surface modifier present in the composition may be, for example, from about 0.001% to about 5% by weight (eg, about 0.01% to about 5% by weight, or about 0.1% by weight to 5% by weight). In some embodiments, the PDI of the composition is about 0.1 or greater and about 0.5 or less (eg, about 0.3 or less, or about 0.2 or less). Methods of use and / or delivery of such compositions to patients or subjects (eg, to mucus or mucous membranes) are also provided.

In one set of embodiments, a composition comprising a plurality of coated particles is provided. The coated particles contain solid medicines or salts thereof, or core particles formed from them, the medicines or salts thereof are no more than about 1 mg / mL at 25 ° C at any point in the pH range (eg,). , About 0.1 mg / mL or less at 25 ° C) and constitutes at least about 80% by weight of core particles (eg, at least 90% by weight, at least about 95% by weight, at least about 99% by weight). .. The coated particles also comprise a coating comprising a surface modifier that surrounds the core particles, the surface modifier being a poloxamer with a hydrophilic block-hydrophobic block-hydrophilic block arrangement, where the poloxamer is Pluronic ( Select from registered trademarks) P123, Pluronic® P103, Pluronic® P105, Pluronic® F127, Pluronic® F108, and combinations thereof, the hydrophobic block with the surface of the core particles. Associating (eg, by adsorption), hydrophilic blocks are present on the surface of the coated particles to make the coated particles hydrophilic, and the surface modifier is at least about 0.001 molecules / nm on the surface of the core particles.²(For example, at least about 0.01 molecule / nm²) Exists. The coated particles have a relative velocity greater than 0.5 in mucus. The coated particles have an average size of at least about 20 nm and about 1 μm or less (eg, about 500 nm or less). The coating thickness may be, for example, about 50 nm or less (eg, about 30 nm or less, about 10 nm or less). The composition may be a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, additives, and / or diluents. In some embodiments, the plurality of coated particles comprises one or more free poloxamers in the composition (eg, Pluronic® P123, Pluronic® P103, Pluronic® P105, It exists in the form of a solution (eg, an aqueous solution) with Pluronic® F127, Pluronic® F108, and a combination thereof. The free poloxamer in solution and the poloxamer on the particle surface may be the same poloxamer and can be present in equilibrium with each other in the composition. The total amount of poloxamer present in the composition is, for example, between about 0.001% by weight and about 5% by weight (eg, between about 0.01% by weight and about 5% by weight, or about 0.1% by weight and about 5% by weight). Between) is fine. In some embodiments, the PDI of the composition is about 0.1 or greater and about 0.5 or less (eg, about 0.3 or less, or about 0.2 or less). Methods of use and / or delivery of such compositions to patients or subjects (eg, to mucus or mucous membranes) are also provided.

In certain embodiments described above, the poloxamer is selected from Pluronic® P123, Pluronic® P103, Pluronic® P105, Pluronic® F127, and combinations thereof. In some embodiments, the poloxamer is not Pluronic® F68 or Pluronic® F108.

These other embodiments of the invention are intended to illustrate specific individual embodiments of the invention, but are not intended to limit the scope as defined in the claims below. Will be further recognized in consideration of.

<p>(Example 1) A non-limiting example of a method for forming non-polymer solid particles into mucous-penetrating particles will be described below. Pyrene, which is a hydrophobic natural fluorescent compound, was used as core particles and prepared by the nano-grinding method in the presence of various stabilizers. The stabilizer served as a surface modifier and formed a coating around the core particles. Various stabilizers / surface modifiers were evaluated to determine the effectiveness of the coated particles that penetrate the mucus.</p><p>Pyrene is nanoground in the presence of various stabilizers / surface modifiers in an aqueous dispersion so that certain stabilizers / surface modifiers 1) promote the reduction of particle size to hundreds of nanometers, 2 ) Whether the surface of the nanoparticles obtained by a mucus inert coating that minimizes the interaction of mucus components with the particles and prevents mucus adhesion can be physically coated (by non-covalent bonds). Judged. In these experiments, stabilizers / surface modifiers acted as a coating around the core particles, and the resulting particles were tested for their mobility in mucus, but in other embodiments, Stabilizers / surface modifiers can be replaced with other surface modifiers that can increase the mobility of the particles in the mucus. Stabilizers / surface modifiers tested were pharmaceuticals such as poly (ethylene oxide) -poly (propylene oxide) -poly (ethylene oxide) block copolymer (Pluronics®), polyvinylpyrrolidone and hydroxypropylmethylcellulose (Methocel). It contained various polymers, oligomers and small molecules shown in Table 2, which contained suitable excipients.</p><p><tables><img file="JP2022017589A_D0004.tif" /></tables><tables><img file="JP2022017589A_D0005.tif" /></tables><tables><img file="JP2022017589A_D0006.tif" /></tables></p><p>An aqueous dispersion containing pyrene and one of the stabilizers / surface modifiers described above was milled with a milling medium until the particle size was reduced to less than 500 nm. Table 3 shows the particle size characteristics of pyrene particles obtained by nanomilling in the presence of various stabilizers / surface modifiers. The particle size was measured by a dynamic light scattering method. When Pluronics® L101, L81, L44, L31, Span20, Span80 or octyl glucoside were used as stabilizers / surface modifiers, stable nanosuspensions could not be obtained. Therefore, these stabilizers / surface modifiers were excluded from further study because they could not effectively promote the reduction in particle size.</p><p><tables><img file="JP2022017589A_D0007.tif" /></tables></p><p>The mobility and distribution of pyrene nanoparticles derived from the nanosuspension produced in human cervical mucus (CVM) was revealed using fluorescence microscopy and multi-particle tracking software. In a typical experiment, a nanosuspension of 0.5 uL (diluted to about 1% of detergent concentration, if necessary) was added to 20 μL of fresh CVM with control. Ordinary nanoparticles (200 nm yellow-green fluorescent carboxylic acid modified polystyrene microspheres from Invitrogen) were used as a negative control to confirm the barrier properties of the CVM sample. Red fluorescent polystyrene nanoparticles coated with PEG5 kDa by covalent bonds were used as positive controls with stable MPP behavior. Using a fluorescence microscope equipped with a CCD camera, each sample of each type of particle, ie, several areas within the sample (pyrene), negative control and positive control, at 100x magnification for 66.7 msec. A 15 second video (15 frames per second) was recorded with a time resolution (the natural blue fluorescence of pyrene allowed the pyrene nanoparticles to be observed separately from the control). Next, using advanced image processing software, the individual trajectories of multiple particles were measured over a time scale of at least 3.335 seconds (50 frames). The obtained transportation data is the trajectory average velocity V._{mean mean}That is, the velocity of the individual particles averaged over its trajectory, and the aggregate average velocity &lt;V._{mean mean}&gt; That is, V averaged over the aggregate of particles_{mean mean}Shown here in the form of. Relative sample velocity &lt;V by equation shown in Equation 1 to allow easy comparison between different samples and to standardize velocity data with respect to natural variation in penetrability of CVM samples._{mean mean}&gt;_relAsked.</p><p>Prior to quantifying the mobility of the pyrene nanoparticles produced, their spatial distribution in mucous samples was evaluated by microscopic examination at low magnification (10x, 40x). It was found that the pyrene / Methocel nanosuspension did not achieve uniform distribution in the CVM and was significantly aggregated in regions much larger than the mucus mesh size (data not shown). Such agglomeration suggests mucus adhesion behavior and effectively prevents mucus invasion. Therefore, further quantitative analysis of particle mobility was considered unnecessary. As with the positive controls, all other experimental pyrene / stabilizer systems achieved a fairly uniform distribution in the CVM. By tracking multiple particles, the negative controls were highly constrained in all test samples, but the positive controls &lt;V._{mean mean}It was confirmed that the positive control was highly mobile, as evidenced by the fact that&gt; was significantly larger than that of the negative control (Table 4).</p><p><tables><img file="JP2022017589A_D0008.tif" /></tables></p><p>Discovered that nanoparticles obtained in the presence of certain (but not all) stabilizers / surface modifiers migrated through the CVM at or near the same rate as the positive control. Was done. Specifically, Pluronics® F127, F108, P123, P105 and P103-stabilized pyrene nanoparticles exceed those of the negative control by about an order of magnitude, as shown in Table 4 and FIG. 2A, and are positive controls. Indistinguishable from the thing within the experimental error range &lt;V_{mean mean}&gt; Was shown. For these samples, &lt;V_{mean mean}&gt;_relThe value was above 0.5, as shown in Figure 2B.</p><p>On the other hand, pyrene nanoparticles obtained with other stabilizers / surface modifiers were 0.4 or less, and most stabilizers / surface modifiers were 0.1 or less, respectively._{mean mean}&gt;_relMost or completely fixed, as demonstrated by the values (Table 4 and Figure 2B). Furthermore, Figures 3A to 3D show V in the particle aggregate._{mean mean}It is a histogram showing the distribution of. These histograms are shown on Pluronic® F127 and Pluronic® F108 in contrast to the mucous adhesion behavior of Pluronic® 87 and Kollidon 25 stabilized samples (selected as representative of mucus-adhesive samples). It shows the mucus diffusion behavior of the stabilized sample (similar histograms were obtained for the Pluronic® P123, P105 and P103 stabilized samples, but not shown here).</p><p>&lt;V_{mean mean}&gt;_relWas mapped to the molecular weight and PEO weight content (%) of the PPO block of Pluronics® used (Fig. 4). It was concluded that PPO blocks of at least 3 kDa and at least those Pluronics® with a PEO content of at least about 30 wt% made the nanocrystals mucus permeable. Although not bound by any theory, hydrophobic PPO blocks are effective with the surface of the core particles when the molecular weight of the PPO block is sufficient (eg, at least about 3 kDa in some embodiments). Hydrophilic PEO blocks are present on the surface of the coated particles when the PEO content of Pluronics® is sufficient (eg, at least 30% by weight in some embodiments). , It is considered that the coated particles can be shielded from the adhesion interaction with mucin fibers. As mentioned herein, in some embodiments the PEO content of the surface modifier is about 10% by weight or more (eg, at least 10% by weight) because the 10% by weight PEO potion makes the particles mucous adherent. It can be selected to be about 15% by weight or at least about 20% by weight).</p><p>(Example 2) In this example, the formation of mucus-penetrating particles using various non-polymer solid particles will be described. To demonstrate the versatility of the approach, the technique described in Example 1 was applied to other non-polymer solid particles. F127 was used as a surface modifier for coating various active pharmaceuticals used as core particles. Sodium dodecyl sulfate (SDS) was selected as the negative control to compare each drug with nanoparticles of similar size of the same compound. Aqueous dispersions containing pharmaceuticals and Pluronics® F127 or SDS were milled with a milling medium until the particle size was reduced to less than 300 nm. Table 5 shows the particle sizes of typical selective agents pulverized using this method.</p><p><tables><img file="JP2022017589A_D0009.tif" /></tables></p><p>To measure the ability of drug nanoparticles to penetrate mucus, we have developed a new assay to measure mass transfer of nanoparticles into mucus samples. Most drugs do not fluoresce spontaneously and are therefore difficult to measure by particle tracking microscopy. The newly developed bulk transport assay does not require the particles to be analyzed to be fluorescent or dye-labeled. In this method, 20 μL of CVM is collected in capillaries and one end is sealed with clay. The open end of the capillary is then immersed in a 20 μL aqueous suspension of 0.5% by weight / volume% drug particles. After the desired time, generally 18 hours, the capillaries are removed from the suspension and the outside is wiped off. Place the capillaries containing the mucus sample in the ultracentrifugation tube. The extraction medium is added to the tube and incubated for 1 hour with mixing, which removes the mucus from the capillaries and extracts the drug from the mucus. The sample is then centrifuged to remove mucin and other insoluble components. The amount of drug in the extracted sample can be quantified using HPLC. The results of these experiments are in good agreement with the results of microscopy, indicating a clear differentiation of transport between mucus-permeable particles and normal particles (CP). Figure 5 shows the transport results of representative selective drugs. These results support microscopic / particle tracking findings for pyrene, demonstrating extension to common active pharmaceutical compounds, and coating of non-polymer solid nanoparticles with F127 increases mucus penetration.</p><p>In Examples 1-2, cervical vaginal mucus (CVM) samples were obtained from healthy female volunteers aged 18 years and older. CVM was collected by inserting a Softcup® menstrual collection cup into the vaginal canal for 30 seconds to 2 minutes as described in the product literature. After removal, CVM was collected from Softcup® by gentle centrifugation at about 30xG to about 120xG in a 50 mL centrifuge tube. In Example 1, CVM was used undiluted and fresh (stored under refrigerated conditions for 7 days or less). Barriers and transport of all CVM samples used in Example 1 were confirmed using negative (200 nm carboxylated polystyrene particles) and positive (PEG5K modified 200 nm polystyrene particles) controls. In Example 2, the CVM was lyophilized and reconstituted. In Example 2, the mucus was frozen at -50 ° C and then lyophilized. The sample was then stored at -50 ° C. Prior to use, the solid was ground into a fine powder using a mortar and pestle and then the mucus was reconstituted by adding water from a final volume equal to the initial volume to a final volume twice the initial volume. The reconstituted mucus was then incubated at 4 ° C for 12 hours and used as described in Example 2. Barriers and transport of all CVM samples used in Example 2 were confirmed using negative (200 nm carboxylated polystyrene particles) and positive (F127 coated 200 nm polystyrene particles) controls.</p><p>(Example 3) This example describes the formation of mucus-penetrating particles using a core containing the drug loteprednol etabonate (LE).</p><p>To demonstrate its usefulness in promoting mucus penetration in the delivery of non-polymer solid particles, it was coated with Pluronic® F127 made by the method described in LE MPP; Example 2 of Loteprednol etabonate MPP formulation. LE particles) were compared with Lotemax®, a currently commercially available formulation. Lotemax® is a steroid eye drop approved for the treatment of ocular surface inflammation. Traditional particles, such as those in Lotemax®, are extensively trapped in the peripheral rapidly-cleared mucus layer in the eye and are therefore also rapidly removed. LE MPP can avoid adhering to the mucus and effectively invade the mucus to promote direct and sustained drug release into the underlying tissue. Improved drug exposure at the labeled site may be able to reduce total doses and increase patient compliance and safety. In vivo, LE to New Zealand White Rabbit A single topical instillation of MPP resulted in significantly higher drug levels in the palpebral conjunctiva, bulbar conjunctiva and cornea compared to the equivalent dose of Lotemax® (FIGS. 6A-6C). At 2 hours, LE levels from MPP were 6, 3 and 8 times higher than those from Lotemax® (eyelids, eyeballs and cornea, respectively). In particular, the level of LE from MPP is about twice as high at 2 hours as the level from Lotemax® at 30 minutes. These results demonstrate the usefulness of the non-polymeric solid MPP method.</p><p>(Example 4) In this example, the formation of mucous permeable particles having a core containing curcumin (CUR) will be described.</p><p>Molecules with various solubilities were selected as model therapeutic agents for forming cored particles of solid pharmaceuticals. One of them, curcumin, has been suggested to have antioxidant, antitumor, and anti-inflammatory properties. It is an interesting candidate due to its high hydrophobicity and natural fluorescence, as well as its widespread potential in medicine. The former feature means that the CUR is poorly soluble in aqueous solution, while the latter allows rapid and unlabeled detection and characterization of particles. The particles were coated with a surfactant (eg, Pluronic® F127, abbreviated as F127 in Examples 4 and 5) to make them mucous permeable.</p><p>In order to formulate CUR particles, we have developed a simple treatment method based on sonication. Briefly, 5 mg of CUR was dispersed in 2 mL of an aqueous solution containing F125 (or other detergent) in a 7 mL vial for scintillation. The suspension was sonicated in a water bath for 20 minutes. The curcumin suspension was then sonicated for 30 minutes using a sonicator with a 3 mm step probe at 100% amplitude. The suspension was centrifuged at 2000 rpm for 10 minutes to remove unbroken crystals. The supernatant was stored at 4 ° C for 2 hours. The supernatant was centrifuged at 16,500 rpm for 20 minutes and then pellets were collected. Without sufficient incubation prior to particle collection, the diffusivity of the coated particles has been shown to be extremely low (data not shown), demonstrating the importance of dense coating at F127 in the formation of mucous permeable particles. Suggests.</p><p>Table 6 and FIGS. 7A-7B summarize the physicochemical properties of the coated CUR particles prepared using the above method. CUR particles (CUR-1% F127 particles) formulated in 1% (weight / volume) F127 possessed an average size of 133 nm, which size was consistent with observation by TEM images (Fig. 7B). .. The zeta potential was close to neutral. CUR particle size and polydispersity (PDI) as the F127 concentration in the CUR suspension during sonication decreases, probably as a result of the decrease in F127 coating density, which leads to a weakening of the stabilizing effect on the CUR particles. Increased respectively (Table 6). There is little effect of the F127 concentration on the zeta potential of the particles, probably because the ionicity of curcumin disappears at pH 4. Powder XRD measurements on CUR-1% F127 particles showed that the chemical structure and crystallinity of the CUR did not change with either sonication or F127 incorporation (Fig. 7A).</p><p><tables><img file="JP2022017589A_D0010.tif" /></tables></p><p>To investigate mucus permeability of CUR-F127 particles, transport of CUR-1% F127 particles was performed in both human cervical and vaginal mucus (CVM) and human cystic fibrosis sputum (CFS) samples. I studied using MPT). Briefly, particles were added to the mucous sample and their motion was recorded using high resolution epi-fluorescence microscopy. Their trajectories and transport rates were then analyzed and quantified. Figures 8A-8B show the time-dependent ensemble mean square geometric mean displacement (&lt;MSD&gt;) of the CUR-1% F127 particles in both CVM and CFS. 200 nm PEGylated (PEG) and carboxylated (COOH) polystyrene (PS) particles were selected as controls for mucus inertness and mucus stickiness, respectively. In all regions of the time scale studied, the &lt;MSD&gt; of CUR-1% F127 particles was comparable to that of PSPEG in CVM and significantly higher than that of PSCOOH in both types of mucus samples. .. On a time scale of 1 second, the &lt;MSD&gt; of CUR-1% F127 particles was 4400 and 220 times larger than that of PSCOOH in CVM and CFS, respectively. CUR-1% F127 Ensemble Geometric Mean Effective Diffusivity on 1 Second Time Scale for Particles (&lt;D_eff&gt;) Was only 1/9 of the theoretically calculated diffusion coefficient in water (Table 6).</p><p>To further investigate the effect of F127 coverage on the transport of CUR particles in mucus, the particles were formulated at various F127 concentrations and their diffusivity was characterized in human CVM (Figure 9, Table 6). ). &lt;D of CUR-0.1% F127 particles and CUR-0.01% F127 particles_eff&gt; Was similar or slightly reduced compared to that of CUR-1% F127 particles, but transport of CUR-0.001% F127 particles was dramatically disrupted (Fig. 9). The diffusivity of CUR-0.001% F127 particles is 1/10000 in human CVM compared to that in water and about 1000 times lower than that of CUR-1% F127 particles in CVM. rice field. In general, reducing the F127 concentration in the preparation of CUR particles results in a decrease in diffusion coefficient, probably because the decrease in F127 concentration reduces the surface density of F127 (and the resulting PEG brush) in equilibrium. It was not clear that this effect was so pronounced when the F127 concentration dropped to 0.01 (weight / volume)%.</p><p><tables><img file="JP2022017589A_D0011.tif" /></tables></p><p>In addition to F127, various Pluronic® were used for pharmaceutical application. Different Pluronic® were tested to see if they functioned equally well or if only certain ones converted CUR particles into mucous permeable particles. Twelve additional Pluronic® as listed in Table 7 (in order of increasing MW of PPO) were selected to prepare the corresponding CUR particles. The resulting size ranged from 90 to 232 nm, most staying between 100 and 150 nm. All types of particles showed higher PDI than that of CUR-1% F127 particles (0.33), most of which were between 0.4 and 0.6. These results mean that F127 may have the strongest stabilizing effect of all Pluronic® tested. The zeta potential was uniformly neutral for all CUR particles studied.</p><p>The transport rates of various Pluronic® coated CUR particles were characterized in human CVM (Table 7). Their diffusivity is that of CUR-1% F127 particles (D)_w/ D_mBy comparing them with = 9), they are divided into three groups: strongly disturbed particle motion (F65, F68; Dw / Dm&gt; 100), disturbed particle motion (F38, F84, F85, F88; 20 Dw). / Dm 100), could be grouped into rapidly penetrating particles (F98, P103, P104, P105, F108, F123; Dw / Dm &lt;20). The fact that all the particles possessed a near-neutral surface charge, but only the formulations with a particular Pluronic® coating exhibited a strongly disturbed diffusivity, with these CUR particles and mucous components. It suggests that the stickiness between them is probably dominated by hydrophobic interactions.</p><p>To identify the factors that determine the diffusivity of CUR particles coated with different Pluronic®, &lt;D of CUR particles_eff&gt; (On a time scale of 1 second) were mapped for the MW of PPO and PEG of Pluronic® used (Figure 10A). &lt;D as the length (Y-axis) of the PPO segment increases_effA general increase in&gt; was observed, but &lt;D_effNo specific pattern was observed between&gt; and PEG MW (X-axis). Surprisingly, this transition occurred at a PPO MW of about 2000 Da, smaller than previously found for polystyrene particles coated with Pluronic®. The mass diffusivity of the particles showed a strong correlation with PPO MW (R = 0.92), but showed little correlation with PEG MW (R = 0.14). It is because Pluronic®, which has a longer PPO segment, has a greater affinity for the hydrophobic surface of CUR particles, so that it is more tightly anchored on the surface for tighter and more stable shielding. It seems to provide.</p><p>To explore the potential of surfactants other than Pluronic® to produce mucus-permeable particles, Tween20, Tween80, and Vitamin E-TPGS, all of which contain PEG segments, were tested. The characteristics of CUR particles prepared in these surfactants are listed in Table 8. All three groups showed a particle size of approximately 150 nm and a near-neutral zeta potential, but their diffusivity was significantly reduced in human CVM compared to water. The slower the transport rate, the less effective the coating of surfactant molecules on the surface of the CUR particles may be, which is attributed to their hydrophobic segments, which are shorter compared to F127. be able to.</p><p><tables><img file="JP2022017589A_D0012.tif" /></tables></p><p>To assess the ability of CUR particles to deliver CUR in a sustained manner, the emission profile of CUR-1% F127 particles was characterized. Briefly, known amounts of CUR particles are suspended in phosphate buffered saline (PBS, pH 7.4) in a 50 mL test tube with a layer of octanol on top to extract the dissolved CUR. Added. The suspension was incubated at 37 ° C with stirring. Octanol was collected and replaced at each point in time. The CUR concentration in octanol was measured by fluorescence analysis. As shown in Figure 11, CUR-1% F127 particles provided continuous release in vitro for 24-48 hours. About 80% of the CUR content was released within the first 24 hours.</p><p>(Example 5) In this example, the development of mucus-permeable particles using the hydrophobic drug 5,10,15,20-tetra (p-hydroxyphenyl) porphyrin (p-THPP) will be described. In addition to CUR, the same method as described in Example 4 was applied to the hydrophobic drug p-THPP. p-THPP is a therapeutic agent used in photodynamic therapy to treat cancer and has been selected as a model photosensitizer in previous studies. Basic properties of p-THPP-1% F127 particles, including size, zeta potential, and diffusivity in human CVM were measured according to the procedure described above (Table 9). Like the CUR-1% F127 particles, the p-THPP-1% F127 particles exhibited a size of 187 nm and a near-neutral surface charge. The diffusivity of p-THPP-1% F127 particles in human CVM is only one-eighth that in water, and the particles are not immobilized by the sticky components in the mucus and are mucus. It has been shown that it can diffuse in the gel at a rate comparable to that of CUR-1% F127 particles.</p><p><tables><img file="JP2022017589A_D0013.tif" /></tables></p><p>(Example 6) This example describes the development of mucus-permeable particles using the highly water-soluble drugs tenofovir (TFV) and acyclovir monophosphate (ACVp).</p><p>Tenofovir (TFV) is a powerful antiviral drug used to treat infectious diseases. Due to the fact that tenofovir (TFV) is highly water soluble, a method for formulating mucous-permeable particles of tenofovir has been developed. The water solubility of TFV is at least 15 mg / mL, so conventional techniques for preparing insoluble particles or encapsulating them in hydrophobic polymeric nanoparticles have not been successful. Interactions between cations and nucleotide / nucleotide analogs were utilized to reduce the water solubility of TFV. TFV interacts very strongly with zinc cations (Zn) via phosphonate groups and purine ring structures. Interactions with zinc can cause TFV sedimentation to result in crystals, stabilizing the crystals with the coatings described herein, stopping agglutination, and measuring the surface properties of the crystals. In addition, Zn is naturally present in vaginal fluid and has recently known antimicrobial properties that have been expanded to include anti-HIV activity.</p><p>Since the crystals and coatings are formed entirely from non-covalent interactions, it is necessary to ensure that the TFV-Zn particles exhibit sustained release in buffer. When compared to a solution of free TFV, the particles showed a much slower release from the 100 kDa dialysis membrane (about 40% after 24 hours).</p><p>The TFV-Zn particles were then formulated using F127 or PVA coating. As can be seen from Table 10, the presence of the coating stabilized the particles, as evidenced by the smaller average size and reduced polydispersity. The presence of a coating on the surface is also indicated by a change towards more neutral charging of the zeta potential. In addition, these particles were fluorescently labeled for contrast purposes by covalent attachment of Alexa Fluor® dye to free amines on TFV. Crystals are unstable when made at a 1:50 labeled: unlabeled TFV ratio, but stable at a 1: 200 labeled: unlabeled TFV ratio, and fluorescence microscopy. It was found that it can be recognized in.</p><p><tables><img file="JP2022017589A_D0014.tif" /></tables></p><p>After obtaining stable fluorescently labeled particles, whether the F127 coating leads to improved particle distribution on the mucosal surface of the animal, as observed consistent with coated polymer nanoparticles. It was judged. TFV particles coated with F127 or PVA were administered into the vagina of the mice, then the mice were sacrificed, their vagina was incised, flattened on a microscope slide and imaged. As can be seen from FIGS. 13A-13B, the F127-coated particles are well distributed over the entire surface of the vagina, while the PVA-coated particles do not penetrate the vaginal folds (mucosal folds). It showed incomplete coverage of the vaginal surface, manifested as "striping" behavior.</p><p><tables><img file="JP2022017589A_D0015.tif" /></tables></p><p>Next, MPP particles were used with acyclovir monophosphate (ACVp) to test the potential efficacy of MPP particles in preventing infection with herpes simplex virus type 2 (HSV-2). Manufactured. ACVp is independent of viral thymidine kinase for the first phosphorylation step that can result in virus resistance. This also imparts anti-HIV activity to ACVp, albeit with low potency.</p><p>Female CF-1 mice aged 6 to 8 weeks were injected subcutaneously with medroxyprogesterone acetate, and one week later, 20 μL of the test drug or PBS was injected into a tip-heat-processed positive displacement capillary pipette (Wiretrol,). It was given intravaginally using Drummond Scientific). After 30 minutes, HSV-2 G strain (ATCC # VR-734, 2.8 × 10) was added to the mouse.⁷TCID₅₀A 10 μL inoculum containing / mL) was loaded. ID, which is the amount of HSV-2 diluted 10-fold with Bartel medium to typically infect approximately 85% of control mice.₅₀Delivered 10 times the amount of. Mice were evaluated by culturing PBS vaginal lavage fluid on human foreskin fibroblasts (Diagnostic Hybrids, MRHF lot number 440318W) for infection 3 days after inoculation (RA Cone, T. Hoen, X. Wong, R. Abusuwwa, DJ Anderson, TR Moench, Vaginal microbicides: detecting toxicities in vivo that paradoxically increase pathogen transmission. BMC Infect Dis 6, 90 (2006)). In this model, the administered (loaded) virus can no longer be detected in the lavage fluid if it is collected more than 12 hours after loading.</p><p>It has been found that ACVp particles can be prepared via interactions similar to TFV (phosphate groups and purine rings) in the presence of zinc. Mice were administered soluble ACVp or ACVp in the form of MPP nanocrystals 30 minutes prior to loading the viable HSV. Both the drug and the virus were administered intravaginally. Soluble drugs given at the same concentration as the MPP drug were ineffective (84% were infected compared to 88% of controls), but only 46.7% of the mice in the MPP drug group were infected. Mice given 10 times the dose of MPP drug were infected at a rate of 62% (drug in PBS) or 69.3% (drug in water). When the soluble drug was compared to the MPP-drug administered in the same medium (pure water), the soluble drug was less protected even at concentrations 10-fold higher than the MPP-drug (p = 0.02).</p><p>It is noted that stable nanocrystals of drugs with free phosphate groups can be formed with zinc using both lyophilization and sonication. In the case of lyophilization, the drug was dissolved in an aqueous solution of F127. The amount of zinc acetate was added in the range of 1:50 to 1: 5 (Zn: drug) and the solution was snap frozen immediately. The dry powder was reconstituted in water at the desired concentration. Stable TFV nanocrystals can be made using both F127 concentrations above (1%) and below (0.08%) the critical micelle concentration (CMC) of F127. However, ACVp nanocrystals were more sensitive to the detergent concentration. Stable nanocrystals could only be formed using F127 concentrations below CMC (about 0.1%). F127 concentrations above CMC caused significant aggregation and precipitation of ACVp nanocrystals, regardless of the pharmaceutical method tested.</p><p>In addition, stable nanocrystals can be formed by adding excess zinc acetate to the drug solution. The precipitate is washed 3+ times by centrifugation. The resulting slurry is sonicated using a probe sonicator (without detergent, foaming results in aggregation and instability). Other methods such as grinding may also work. A surfactant is then added to stabilize the resulting nanocrystals. In the absence of surfactant, significant aggregation and sedimentation occur. Using this formulation technique, stable TFV nanocrystals could be made using F127 concentrations, eg, up to 1%. Similarly, in these experiments, stable ACVp nanocrystals could only be formed using F127 concentrations below CMC (typically 0.08%).</p><p>In Examples 4-6, undiluted cervical vaginal discharge from a woman with a normal vaginal plexus using a self-sampled menstrual collection device, a protocol approved by the Johns Hopkins University Trial Review Board. Collected according to. The instrument was inserted into the vagina for approximately 30 seconds, removed and placed in a 50 mL centrifuge tube. The sample was centrifuged at 1,000 rpm for 2 minutes to collect mucus secretions.</p><p>In Examples 4-6, particle transport rates are measured by analyzing the trajectory of fluorescent or fluorescently labeled particles and are inverted with a 100x oil immersion objective (NA, 1.46) and a suitable filter. Recording was performed using an electron double charge coupled device (EMCCD) camera (Evolve512, Photometrics, Tucson, Arizona) mounted on an epi-microscope (Zeiss, Thornwood, New York). Experiments were performed in a custom chamber slide with diluted particle solution (0.0082 (weight / volume)%) in 20 μL fresh mucilage to a final concentration of 3 (volume / volume)% (final particle concentration, 8.25 ×). 10-7 weight / volume) and stabilized at room temperature prior to microscopic observation. Trajectories of n 100 particles were analyzed for each experiment, and at least 3 independent experiments were performed for each condition. Video to Meta Morph Software (Universal Imaging) , Glendale, Wisconsin), captured for 20 seconds with a time resolution of 66.7 ms. The tracking resolution was 10 nm as measured by tracking the displacement of the particles immobilized with strong adhesion. The coordinates of the center of gravity of the nanoparticles are &lt;Δr2 (τ)&gt; = [x (t + τ) -x (t)].²+ [y (t + τ)-y (t)]²Converted to a time average MSD calculated as. Where x and y represent the coordinates of the nanoparticles at a given time, and τ is a time scale or time lag. The distribution of MSD and the effective diffusivity were calculated from this data. Particle penetration into the mucus layer was modeled using Fick's second law and the diffusion coefficient obtained from follow-up experiments.</p><p>(Example 7) This example describes the development of mucous permeable particles that improve drug delivery to the mucosal surface of the vagina of mice.</p><p>Improved methods for sustained and more uniform drug delivery to the vagina are more effective for conditions that have a detrimental effect on women's health, such as cervical cancer, bacterial vaginosis, and sexually transmitted diseases. May provide prophylactic and therapeutic treatment. For example, women are disproportionately infected with HIV due to the lack of female-led preventive measures to some extent. Easily administered, individualized and effective methods to protect women from vaginal HIV transmission may prevent millions of infections worldwide. However, vaginal folds or "mucosal folds" that adapt to sexual intercourse and extension during labor are typically crushed by intra-abdominal pressure, making the surface of these folds less accessible to the drug and drug carrier. Inadequate distribution in the vaginal folds has been mentioned as a decisive factor for the failure to protect the susceptible vaginal surface from infection, even after simulated sexual intercourse. Distribution over the surface of susceptible targets has been found to be important for the prevention and treatment of infectious diseases. In addition, the drug delivered to the vagina should be retained in the vaginal canal at an effective concentration for a long period of time in order to increase the tolerability of the user. Achieving sustained local drug concentrations is due to the high permeability of the vaginal epithelium to small molecules, and soluble drug dosage forms (gels, creams) are eliminated by intra-abdominal pressure and gait. It's a difficult task because it can happen. Finally, the drug delivery method must be safe and non-toxic to the vaginal epithelium. Improvements in the distribution, retention, and safety profile of vaginal dosage forms substantially increase efficacy and reduce side effects caused by systemic treatments that are not sufficiently effective against cervical infections and diseases. May lead to.</p><p>Nanoparticles have received considerable attention due to their ability to provide sustained topical drug delivery to the vagina. However, the mucous layer covering the vaginal epithelium presents a barrier to achieving uniform distribution and long retention in the vaginal canal. Mucus efficiently captures most particles, including conventional polymeric nanoparticles (CPs), through both sticky and steric interactions. The effectiveness of mucus in capturing foreign pathogens and particles leads to immediate capture of the CP upon contact with the luminal mucus layer, preventing penetration into the mucosal folds and thus hindering protection of the mucosal folds. means. Particles and pathogens trapped in the luminal surface mucous layer are expected to be rapidly eliminated from the tissue, limiting the retention time of mucous adhesive materials such as CP.</p><p>By simulating a virus that has recently evolved to penetrate the mucous barrier to colonize the infection, mucus permeable particles (MPP) have a very high density of low CP for drug delivery to the mucosa. Made by coating with molecular weight poly (ethylene glycol) (PEG). MPPs diffuse in human cervical mucus (CVM) at a rate comparable to their theoretical diffusion in water. In this case, MPP provides enhanced distribution and increased retention in the vagina in vivo by penetrating into the deepest mucus layer containing mucus that is more slowly eliminated in the mucosal folds. It was sought to test the hypothesis that the drug would be released to the optimal location for efficient tissue uptake (Fig. 14E). In addition to the general progestin-induced estrous cycle (DP) mouse model, mouse CVM (mCVM) more closely mimics human CVM (hCVM), thus developing and migrating MPPs for human use. Therefore, the use of the estradiol-induced estrous cycle (IE) mouse model, which provides a more human-like model, is introduced.</p><p>Carboxylic acid-coated fluorescent polystyrene nanoparticles (PS-COOH) were converted to MPP by covalently bonding a dense coating of low molecular weight PEG, as previously reported (YY Wang,). SK Lai, JS Suk, A. Pace, R. Cone, J. Hanes, Angew Chem Int Ed Engl 47, 9726-9729 (2008); SK Lai, DE O'Hanlon, S. Harrold, ST Man, YY Wang, R. Cone, L. Hanes, Proc Natl Acad sci USA 104, 1482-1487 (2007)). In addition, biodegradable particles can be loaded with drugs and are suitable for administration to humans, so biodegradable MPP (BD-MPP), as previously reported, is poly ( Formulated using a core of lactic acid-co-glycolic acid (PLGA) and a physically adsorbed PEG coating (M. Yang, SK Lai, YY Wang, W. Zhong, C. Happe, M. Zhang, J. Fu, J. Hanes, Biodegradable Nanoparticles Composed Entirely of Safe Materials that Rapidly Penetrate Human Mucus. Angew Chem Int Ed Engl 50, 2597-2600 (2011)). PS-COOH and PLGA nanoparticles have a highly negative surface charge that is almost neutralized when densely coated with PEG. The nanoparticles were determined to be well covered by measuring the zeta potential as previously described (Table 12) (SK Lai, DE O'Hanlon, S. Harrold, ST Man, YY. Wang, R. Cone, J. Hanes, Proc Natl Acad Sci USA 104, 1482-1487 (2007)). It has previously been found that zeta potentials above -10 mV are essential for mucus permeation properties in hCVM (YY Wang, SK Lai, JS). Suk, A. Pace, R. Cone, J. Hanes, Addressing the PEG mucoadhesivity paradox to engineer nanoparticles that "slip" through the human mucus barrier. Angew Chem Int Ed Engl 47, 9726-9729 (2008)). Particles were administered intravaginally to estrus mice to ensure that MPP was mucus permeable in the original estrus mCVM. The entire vagina was then removed and the multiparticle tracking (MPT) method (J. Suh, M. Dawson, J. An incision was made in Hanes, Adv Drug Deliv Rev 57, 63-78 (2005) to visualize the motion of hundreds of individual particles. The particle trajectory for MPP hinted at rapid diffusion in the water-rich pores of the mCVM, while the motion of uncoated PS-COOH nanoparticles (CP) It was smaller than the particle diameter (about 100 nm) (Fig. 14A). The mean squared displacement (&lt;MSD&gt;) of the MPP in mCVM is comparable to that reported for MPP in hCVM (SK Lai, YY Wang, K. Hida, R. Cone, J. Hanes, Nanoparticles). P Natl Acad Sci USA 107, 598-603 (2010)) (Fig. 14B), theoretical diffusion of 110 nm particles in water (about 4 μm).²Ensemble average effective diffusion coefficient (&lt;D), which is only about one-eighth of (/ sec)_eff&gt;) Was found to correspond.</p><p><tables><img file="JP2022017589A_D0016.tif" /></tables></p><p>D measured for individual particles_effBased on, about half of the MPPs were evaluated by Fick's second law of diffusion, which diffuses through a 100 μm-thick layer of mCVM in about 4 hours (Fig. 14D), while in CP after 24 hours. Even there was no perceptible penetration. D on a 1 second time scale for CP_effThe values correspond to MSD values below the particle diameter (dotted line, Figure 14C), probably indicating thermal fluctuations of the particles colliding with the mucin fibers rather than diffusion of the particles. Overall, the transport behavior of MPPs and CPs in estrus mCVM was very similar to their transport behavior in hCVM.</p><p>Synchronizing a large number of mice in estrus for retention studies required hormonal treatment. Endocrinology 34, 269-275 (1944); Endocrinology 34, 269-275 (1944); And CA Rubio, The exfoliating cervico-vaginal surface. II. Scanning electron microscopical studies during the estrous cycle in mice. Particle transport behavior was tested in IE mice to confirm that Anat Rec 185, 359-372 (1976)) did not alter MPP and CP transport behavior prior to distribution and retention studies (Fig. 15A). .. Furthermore, the transport behavior of BD-MPP was indistinguishable from MPP in IE mucus (Fig. 15B).</p><p>Next, we investigated whether the ability to rapidly penetrate mucus in estrus and IE mice led to a more rapid and uniform distribution of MPP in the vagina compared to CP. MPPs and CPs were applied into hypopermeable media to simulate how infiltration-driven water flow (advection transport) rapidly transports nutrients from the intestinal lumen to the brush border epithelial surface. Ten minutes after particle administration, the entire vagina was removed and stained for cell nuclei. CP aggregated in the luminal mucus and did not penetrate into the vaginal mucosal folds (Fig. 16). In contrast, MPP formed a continuous particle layer covering all vaginal epithelium, including all surfaces of mucosal folds. MPP penetrated into mucus greater than about 100 μm via advection within 10 minutes compared to the approximately 4 hours required to diffuse that distance in the mucus (Fig. 14D). This behavior was also consistent with BD-CP and BD-MPP administered into IE mice, as well as CP and MPP (Fig. 16). Videos showing the movement of MPP beyond the mucous sticky CP in hCVM can be found in Video 1 (no flow, diffusion) and Video 2 (with flow, advection).</p><p>Fluorescent images of newly excised, incised and flattened mouse vaginal tissue were obtained to quantify differences in MPP and CP distribution. As can be seen from FIG. 17, the adhesion of CP to the luminal mucus layer of the vagina creates "stripes" of particle-containing mucus, which alternate with the dark "stripes" of particle-free mucus. It corresponds to the mucosal folds that are opened when the vaginal tissue is flattened. In contrast, transport of MPP towards the epithelium and into the mucous folds created a continuous particle coating on the flattened vaginal surface (Fig. 17). Quantification of fluorescence on vaginal and cervical tissue shows that 88% of the flattened vaginal surface and 87% of the cervical surface are densely covered with MPP, while only 30% of the vaginal surface and the uterus. It was shown that 36% of the cervical surface was covered with CP. Further observations at higher magnifications of darker areas of the vaginal and cervical surface show a less concentrated continuous coating of MPP (Figs. 17-18, inset) and the vagina and cervix are nearly complete. It means that it was covered with. In the case of CP, no coating was found at higher magnification and less concentrated (Figs. 17-18, inset). Similar trends were found, with BD-MPP having 85% vaginal coverage and 86% cervical coverage, and BD-CP having 31% vaginal coverage and 27% cervical coverage. (Figs. 17-18).</p><p>It was then sought to determine if improved distribution of BD-MPP could improve delivery of small molecules compared to gel dosage forms. Lipophilic molecules probably invade the first epithelial surface they come into contact with and cannot contact the cells in the mucosal folds. Conversely, hydrophilic molecules can diffuse rapidly in the vaginal epithelium and are carried away by blood and lymph circulation, leading to short-term coverage. BD-MPP was loaded with fluorescein isothiocyanate (FITC), a fluorescent water-soluble small molecule, as a model drug (FITC / MPP). To simulate conventional vaginal delivery, soluble FITC (FITC / gel) was added to hydroxyethyl cellulose (HEC), a general purpose vaginal placebo gel. Twenty-four hours after administration to estrous mice, vaginal tissue was removed and flattened to expose vaginal folds. The FITC patch covers 42% of the vaginal surface when administered as a FITC / gel, while the FITC / MPP provides a well-retained FITC coating on the vaginal surface even 24 hours after particle administration. Provided to 87% of.</p><p>To further characterize the effect of the mucus barrier, we can remove vaginal mucus by washing prior to administration of the particles (Y. Cu, CJ Booth, WM Saltzman, In vivo distribution of surface-modified PLGA nanoparticles). following intravaginal delivery. Journal of Controlled Release 156, 258-264 (2011); and KA Woodrow, Y. Cu, CJ Booth, JK Saucier-Sawyer, MJ Wood, WM Saltzman, Intravaginal gene silencing using biodegradable polymer nanoparticles densely loaded with small -interfering RNA. Nat Mater 8, 526-533 (2009)) found that they significantly improved the distribution of CPs and showed that their mucous adhesion properties interfered with uniform distribution in the vagina (Fig. 19).</p><p>Next, it was sought to use our IE model to measure vaginal retention of MPP in comparison to mucous sticky CP. Fluorescent MMP and CP were administered intravaginally to IE mice. At the designated time point, the entire genital tract (vagina and uterine horn) was removed and quantitatively analyzed by fluorescence imaging (Fig. 21A). After an initial reduction in particle fluorescence that was similar for MPP and CP (perhaps due to an initial "squeeze out" prior to mucus penetration), MPP residuals remained constant at approximately 60%. (Fig. 21B). In contrast, the amount of CP steadily decreased to 10% (6 hours) over time. Importantly, CP was distributed along the length of the vagina, and this lengthwise coverage is as shown in Figure 16 where CP penetrates the mucus into the epithelium as well as the vagina. Indicates that it does not reach the inner surface of the folds.</p><p>The immune system is highly active on mucosal surfaces, especially those with surfaces covered with viable cells such as columnar epithelium in the human endometrium (OP Mestecky J, McGhee JR, and Lambrecht BN, Mucosal). Immunology. (Elselvier Academic Press, Burlington, ed, Third, 2005)). Using mice pretreated with Depo-Provera, a long-acting progestin treatment that synchronizes the inflammatory effects of nanoparticles during the resting phase of the mice, during which the vaginal epithelium becomes thin and covered with viable cells. I checked it. In contrast, during estrus, the mouse vagina thickens from 4-7 cell layers to about 12 cell layers, and the epithelial surface is protected by many layers of dead and drying cells (Biology of the laboratory). mouse. Edited by George D. Snell, Dover Publications, Inc., New York, 1956 (Reprint of first edition, 1941). Journal of the American Pharmaceutical Association 45, 819-819 (1956)). In addition, the vaginal epithelium of progestin-induced estrous cycle (DP) mice has an increased immune cell population, leading to an increased acute inflammatory response, while the estrous cycle is characterized by the absence of immune cells (CH Hubscher). , DL Brooks, JR Johnson, A quantitative method for assessing stages of the rat estrous cycle. Biotech Histochem 80, 79-87 (2005)). Depo-Provera effectively synchronizes mice to estrus-like periods for days to weeks, which are important for experiments that last for more than 24 hours.</p><p>Standard hematoxylin and eosin (H &amp; E) staining was used to investigate the potential toxic effects of nanoparticles administered intravaginally. Known to cause vaginal toxicity (G. Ramjee, A. Kamali, S. McCormack, The last decade of microbicide clinical trials in Africa: from hypothesis to facts. AIDS 24 Suppl 4, S40-49 (2010)) Nonoxynol-9 (N9), a nonionic detergent, was used as a positive control and PBS (physiological saline) was used as a negative control. The same (BD-) MPP and (BD-) CP used in distribution and retention studies were tested for toxicity. As expected, N9 caused acute inflammation that was not observed after PBS treatment at 24 hours (Fig. 23). CP, like N9, caused overt neutrophil infiltration into the lumen, but MPP did not cause this inflammatory effect (Figure 23, arrow).</p><p>Recent studies have shown that vaginal epithelium can secrete immune mediators that may increase susceptibility to sexually transmitted infections in response to specific vaginal products (JE Commins, Jr.). ., GF Doncel, Biomarkers of cervicovaginal inflammation for the assessment of microbicide safety. Sex Transm Dis 36, S84-91 (2009); and SS Wilson, N. Cheshenko, E. Fakioglu, PM Mesquita, MJ Keller, BC Herold, Susceptibility to genital herpes as a biomarker predictive of increased HIV risk: expansion of a murine model of microbicide safety. Antivir Ther 14, 1113-1124 (2009)). Therefore, it is important that vaginal products do not elicit such an immune response, especially after repeated doses. Since our ultimate goal was to test MPP for protection against HSV-2, we have introduced MPP formulations containing acyclovir monophosphate (ACVp) to recent tenofovir. Compared with N9, HEC placebo gel, PBS, and gel medium (TFV medium) used in clinical trials. Nanoparticles and control formulations, Depo Mice treated with Provera were administered transvaginally daily for 7 days. Vaginal lavage fluid was collected from each mouse on day 8 and was found to be enhanced in response to epithelial stimulation Cytokines: i.e., interleukin 1β (IL-β), interleukin 1α (IL-1α). , Tumor necrosis factor α (TNF-α), and interleukin 6 (IL-6) were evaluated. It was found that both IL-1α and IL-1β levels were elevated in response to both the TFV medium and the N9 solution (Fig. 24). This was not surprising in the case of N9 treatment, considering that IL-1α and IL-1β are secreted by the vaginal epithelium in response to injury (JE Cummins, Jr., GF Doncel, Biomarkers of). cervicovaginal inflammation for the assessment of microbicide safety. Sex Transm Dis 36, S84-91 (2009)). In contrast, cytokine levels associated with ACVp-MPP were comparable to those associated with HEC placebo gels used in clinical trials without any increase in susceptibility to infection (Fig. 24) (QA Karim,). SSA Karim, JA Frohlich, AC Grobler, C. Baxter, LE Mansoor, ABM Kharsany, S. Sibeko, KP Mlisana, Z. Omar, TN Gengiah, S. Maarschalk, N. Arulappan, M. Mlotshwa, L. Morris, D . Taylor, CT Grp, Effectiveness and Safety of Tenofovir Gel, an Antiretroviral Microbicide, for the Prevention of HIV Infection in Women. Science 329, 1168-1174 (2010); and D. Tien, RL Schnaare, F. Kang, G. Cohl, TJ McCormick, TR Moench, G. Doncel, K. Watson, RW Buckheit, MG Lewis, J. Schwartz, K. Douville, JW Romano, In vitro and in vivo characterization of a potential universal placebo designed for use in vaginal microbicide clinical trials. AIDS Res Hum Retroviruses 21, 845-853 (2005)). There was no detectable increase in IL-6 or TNF-α associated with any vaginal treatment compared to untreated controls.</p><p>Finally, we investigated whether improved distribution, retention, and toxicity profiles of MPPs lead to improved protection against vaginal HSV-2 loading in mice. Depo Provera treatment significantly increased the vaginal susceptibility of mice to infections, and candidate bactericides were partially protected in the mouse model used here, even when administered immediately prior to the infectious inoculum. (SL Achilles, PB Shete, KJ Whaley, TR Moench, RA Cone, Microbicide efficacy and toxicity tests in a mouse model for vaginal transmission of Chlamydia trachomatis. Sex Transm Dis 29, 655-664 (2002) ; And L. Zeitlin, KJ Whaley, TA Hegarty, TR Moench, RA Cone, Tests of vaginal microbicides in the mouse genital herpes model. Contraception 56, 329-335 (1997)). In addition, some vaginal product excipients actually increase susceptibility to infection in this model (RA Cone, T. Hoen, X. Wong, R. Abusuwwa, DJ Anderson, TR Moench, Vaginal microbicides). : detecting toxicities in vivo that paradoxically increase pathogen transmission. BMC Infect Dis 6, 90 (2006); and TR Moench, RJ Mumper, TE Hoen, M. Sun, RA Cone, Microbicide impurities can greatly increase susceptibility to genital herpes transmission in the mouse. BMC Infect Dis 10, 331 (2010)). Acyclovir was selected to test ACVp for blocking vaginal infections with HSV-2 infections because it provides virus suppression in animals with multiple daily doses (ER Kern, Acyclovir Treatment of Experimental). Genital Herpes-Simplex Virus-Infection. Am J Med 73, 100-108 (1982)). However, a single vaginal pretreatment with 50 mg / mL (5%) ACVp in guinea pigs resulted in infection in 70% of animals compared to controls (ER Kern, J. Palmer, G. Szczech, G. Painter). , KY Hostetler, Efficacy of topical acyclovir monophosphate, acyclovir, or penciclovir in orofacial HSV-1 infections of mice and genital HSV-2 infections of guinea pigs. Nucleos Nucleot Nucl 19, 501-513 (2000)). Therefore, ACVp provides a study case to determine if MPP could significantly improve water-soluble, rapidly metabolized drug protection by prolonging treatment-related drug concentrations after a single dose. did. In addition, the mechanism of action of nucleotide analogs such as ACVp is the disruption of intracellular viral replication, where successful protection means efficient uptake and retention in susceptible target cell populations in the vaginal and cervical mucosa. be.</p><p>ACVp nanoparticles were formulated with the same mucus-inert coating used for all other studies. The size and zeta potential of ACVp-MPP were similar to polystyrene (PS) based MPP (Table 12). Mice were intravaginally administered soluble ACVp or ACVp-MPP 30 minutes prior to loading HSV-2. Soluble drugs administered at the same concentration as ACVp-MPP (1 mg / mL) were ineffective in protecting mice from viral infection (84.0% infected compared to 88% of controls), while , Only 46.7% of mice in the ACVp-MPP group were infected (Table 11). The group of mice given 10 times the concentration of the soluble drug in ACVp-MPP was also infected at a rate of 62.0% (drug in PBS) or 69.3% (drug in water). When the soluble drug was compared to ACVp-MPP in the same medium (pure water), the soluble drug was significantly less protective, even at concentrations 10-fold higher than ACVp-MPP (Table 11). 6-8 week old CF-1 mice (Harlan) were used to study the distribution and retention of nanoparticles on the vaginal mucosal surface and the effect of repeated doses. Mice were housed in a reverse light cycle facility (12 hours light / 12 hours dark). In the case of natural estrous cycle, mice were selected for external estrous cycle and confirmed by close scrutiny (AK Champlin, DL Dorr, AH Gates, Determining the stage of the estrous cycle in the mouse by the appearance of). the vagina. Biol Reprod 8, 491-494 (1973); E. Allen, The oestrous cycle in the mouse. American Journal of Anatomy 30, 297-371 (1922)). In the case of hormone-induced estrus (IE), mice were acclimatized for 3 weeks and 100 μg of benzoate 17-β estradiol (Sigma) was injected subcutaneously for 2 days prior to the experiment. Many studies have demonstrated that treatment with estradiol induces an "estrus-like" state with similar epithelial properties and vaginal cell populations (CG Rosa, JT Velardo, Histochemical localization of vaginal oxidative enzymes and mucins). Ann NY Acad Sci 83, 122-144 (1959); CA Rubio, The exfoliating cervico-vaginal surface, II. Scanning electron microscopical studies during the estrous cycle in mice. Anat Rec 185, 359- 372 (1976); AE Gillgrass, SA Fernandez, KL Rosenthal, C. Kaushic, Estradiol regulates susceptibility following primary exposure to genital herpes simplex virus type 2, while progesterone induces inflammation. Journal of Virology 79, 3107-3116 (2005)). For vaginal toxicity and cytokine release, 2.5 mg of Depo-Provera (medroxyprogesterone acetate, 150 mg / mL) (Pharmacia &amp;) was given to mice prior to the experiment.</p><p>Water was used as the hypotonic medium for all particle solutions. For in vitro follow-up, 5 μL particles were administered intravaginally. Approximately 10 minutes later, the vagina was removed, carefully cut open and flattened. The entire tissue was placed in a custom-made, well-structured well configured to allow the cover glass to be placed on the top surface and contacted with mucus without deforming the tissue. The wells were approximately 1 mm x 0.5 mm rectangles cut out from three layers of insulating tape adhered to a standard glass slide. Around the edge of the cover glass was sealed with superglue and immediately contrasted to prevent drying.</p><p>Mice were anesthetized prior to experimental procedures involving sacrifice due to cervical dislocation. In all studies, mice were protected from self-grooming by a brim of weakly adhesive tape around the abdomen and from mutual grooming by accommodating them in individual cages.</p><p>For conventional mucous adhesive particles (CP), 100 nm size fluorescent polystyrene (PS) nanoparticles (PS) nanoparticles modified with carboxyl (COOH) were used. These particles are characterized by a negatively charged surface at neutral pH (Table 12). To produce mucus-penetrating particles (MPP), CP was subjected to amine-modified 5 kDa PEG (Creative PEG works) by a standard 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide coupling reaction. Modified by covalent bond. Grain size and ζ potential were measured by dynamic light scattering and laser Doppler flow velocity measurement methods using Zetasizer Nano ZS90 (Malvern Instruments), respectively. Size measurements were performed at a scattering angle of 25 ° C and 90 °. Samples were diluted with 10 mM NaCl solution (pH 7) and measurements were performed according to the equipment instructions. The PEG composite was confirmed using the near-neutral ζ potential measured by the laser Doppler flow velocity measurement method.</p><p>PLGA acid 2A (50:50 Lakeshore Biomaterials), Lutrol F127 (BASF), and poly (vinyl alcohol) (PVA 25kDa, Polysciences) were used as biodegradable particles. Alexa Fluor555 was chemically complexed with PLGA used to produce nanoparticles by nanoprecipitation as previously described (M. Yang, SK Lai, YY Wang, W. Zhong, C. Happe, M. Zhang, J. Fu, J. Hanes, Biodegradable Nanoparticles Composed Entirely of Safe Materials that Rapidly Penetrate Human Mucus. Angew Chem Int Ed Engl 50, 2597-2600 (2011)). Briefly, 10 mg / mL labeled PLGA was dissolved in acetone or THF (with or without 2 mg of FITC) and added dropwise to 40 mL of aqueous detergent solution. After stirring for 2 hours, the particles were filtered through a 5 μm syringe filter (Sorvall RC-6 +, Thermo Scientific), collected by centrifugation and washed. The particle size and zeta potential were measured as described.</p><p>ACVp-MPP was prepared by dissolving ACVp in ultrapure water containing Lutrol F127. Zinc acetate was added in a molar ratio of ACVp: Zn of 5: 1 to chelate ACVp, making it water-insoluble, then immediately quick-frozen and lyophilized. Particle characterization was performed after reconstruction. Prior to administration, the powder was reconstituted with ultrapure water to a final concentration of 1 mg / mL ACVp and 0.8 mg / mL Lutrol. Soluble ACVp was titrated with NaOH as required to bring the pH to 6-7. The particle size and zeta potential were measured as described.</p><p>Silicon-enhanced target cameras (VE-1000, Dage-MTI) mounted on an inverted epi-illumination microscope equipped with a 100 x oil-immersed objective lens (numerical aperture 1.3) to capture the trajectory of fluorescent particles in an in vitro vaginal tissue sample. ) Was recorded. The video was captured for 20 seconds with a time resolution of 66.7 ms using Metamorph software (Universal Imaging Corp.). Trajectories of n&gt; 130 particles were analyzed for each experiment and three independent experiments were performed using tissues from different mice. The coordinates of the center of gravity of the particle are &lt;Δr²(τ)&gt; = [x (t + τ)-x (t)]²+ [y (t + τ)-y (t)]²Converted to the time mean square geometric mean displacement (&lt;MSD&gt;) calculated as.</p><p>In the equation, τ is the time scale (or time lag) and x and y are the coordinates of the corresponding particles at time t, Δr.²Is MSD. This equation, as previously substantiated, is the MSD and effective diffusivity (D) of the particles._eff) Was used to calculate (SK Lai, DE O'Hanlon, S. Harrold, ST Man, YY Wang, R. Cone, J. Hanes, Rapid transport of large polymer nanoparticles in fresh undiluted human mucus. Proc Natl Acad Sci USA 104, 1482-1487 (2007); BC Tang, M. Dawson, SK Lai, YY Wang, JS Suk, M. Yang, P. Zeitlin, MP Boyle, J. Fu, J. Hanes, Biodegradable polymer nanoparticles Proc Natl Acad Sci USA 106, 19268-19273 (2009)). Calculated D_effThe values were used to model particle penetration in the mucus plate as previously described (B, C. Tang, M. Dawson, SK Lai, YY Wang, JS Suk, M. Yang, P. Zeitlin, MP Boyle, J. Fu, J. Hanes, Biodegradable polymer nanoparticles that rapidly penetrates the human mucus barrier. Proc Natl Acad Sci USA 106, 19268-19273 (2009)).</p><p>For distributions with mucus removal, mice were vaginal washed twice with 50 μL PBS followed by a single wipe with a cotton-tipped applicator prior to particle administration. Subsequently, 5 μL of CP or MPP was administered intravaginally. The entire vagina was then removed and frozen in Tissue Tech OCT Compound (Sakura Fine Tech USA). Cross-sections were obtained at various points along the length of the tissue (between the vaginal opening and the cervix) using the Microm HM500M Cryostat (Microm International). Section thickness was set to 6 μm to achieve single cell layer thickness. Then, in order to visualize the cell nucleus and retain the fluorescence of the particles, the section is divided into ProLong containing DAPI. Stained with Gold (Invitrogen) anti-fading encapsulant. Fluorescent images of the sections were obtained using an inverted fluorescence microscope. To quantify the distribution of nanoparticles, 5 μL CP or MPP was administered intravaginally. Within 10 minutes, vaginal tissue was cut thinly in length, including the non-particle-administered "blank" tissue, sandwiched between two glass slides and sealed with superglue. By this treatment, the tissue is completely flattened and the folds are exposed. The "blank" tissue was used to assess the background fluorescence level of the tissue to ensure that all images taken were well above the background level. Six fluorescent images at low magnification and at least one image at high magnification were taken for each tissue. To draw a boundary around the fluorescent signal, the image was thresholded and then the covered area was quantified using ImageJ software. Average coverage was determined for each mouse and then these values were averaged across the group of mice with n 3. The cervix from each mouse was cut from the uterine horn and attached using a custom well similar to that used for in vitro particle tracking. Wells were sealed with cover glass and background fluorescence levels were measured using a blank tissue. A single fluorescent image of almost the entire surface of the cervix was taken at low magnification above the tissue background level. Thresholds were set for these images in a similar manner and the area covered by the particles was measured. At least one higher magnification image was taken for each tissue to show the individual particles.</p><p>MPP and CP advection was visualized using a custom-made capillary tube device. A flat capillary tube (0.4 mm x 4 mm x 50 mm, VitroCom) was attached to a 1 mL tuberculin syringe (Becton Dickinson) via a flexible plastic tube. The tube was attached to one end of the capillary tube and sealed with silicone grease. Fresh, undiluted humans mixed with saline in syringes and tubes, followed by 3% (volume / volume) of approximately 500 nm uncoated (red fluorescent) and PEG-coated (green fluorescent) polystyrene beads (Invitrogen). Loaded CVM. PEG-coated beads were prepared as previously described for the 100 nm MPP used in the mouse study. Approximately 80 μL of mucus was required to fill the capillary tube, and care was taken to avoid the introduction of air bubbles. Time-lapse videos showing the motion of MPPs and CPs in a capillary tube with or without pressure were recorded using a 40x objective on a Zeiss LSM510 confocal microscope (Carl Zeiss MicroImaging, LLC).</p><p>The FITC dye (Sigma-Aldrich) was mixed with a HEC gel courtesy of T. Moench (Reprotect) at 1 mg / mL. Biodegradable MPPs were prepared as described, loaded with FITC dye and suspended in 1% Lutrol F127. A 10 μL gel or particle solution was administered intravaginally to assess distribution. Twenty-four hours later, vaginal tissue was removed and incised to flatten it. The tissue was then sandwiched between two microscope slides and crushed to flatten the mucosal folds. The exposure settings used, including "blank" tissue to determine background autofluorescence from vaginal tissue, ensured the presence of FITC. A Nikon E600 inverted microscope equipped with a 2x objective lens was used to obtain fluorescence images of the dye distribution on the flattened tissue surface. Thresholds were set for these images in a similar manner using ImageJ to measure the coverage area.</p><p>To assess the retention of nanoparticles, 5 μL of red fluorescent CP or MPP was administered intravaginally. All cervical and vaginal ducts were obtained at 0, 2, 4 and 6 hours and placed in standard tissue culture dishes. N&gt; 7 mice were used for each condition and time point. Fluorescent images of the tissue were acquired using the Xenogen IVIS Spectrum Contrast Device (Caliper Life Sciences). The Xenogen Living Image 2.5 software was used to calculate the amount of fluorescence count per unit area.</p><p>5 μL of particles or control solution was administered intravaginally to the DP mouse model. After 24 hours, the entire cervical vaginal canal was obtained and fixed in 4% paraformaldehyde solution for 24 hours. Tissues were placed in 70% ethanol and sent to the Johns Hopkins Reference Histology Laboratory for paraffin embedding and H &amp; E staining.</p><p>Each 20 μL study drug was administered intravaginally to the DP mouse model once daily for 7 days. HEC gels and N9 were provided by T. Moench (Reprotect), and medium gels for TFV were provided in favor of C. Dezzutti (University of Pittsburgh). On day 8, each mouse was washed twice with 50 μL PBS. Each wash sample was diluted with an additional 200 μL PBS and centrifuged to remove mucus plugs. The supernatant (200 μL) was removed and divided into 50 μL for each of the four Quantikine ELISA kits (R &amp; D Systems, Inc.) (IL-1β, IL-1α, TNF-α, and IL-6). ELISA was performed according to the manufacturer's instructions.</p><p>All data are shown as a mean with a standard error (SEM) of the shown mean. Statistical significance was determined by a two-sided Student's t-test (α = 0.05) assuming unequal variance. For HSV-2 loading, statistical significance was determined using Fisher's direct test, bilateral distribution.</p><p>Female reproductive tracts are susceptible to a wide range of sexually transmitted diseases (R. Mallipeddi, LC Rohan, Nanoparticle-based vaginal drug delivery systems for HIV prevention. Expert Opin Drug Deliv 7, 37-48 (2010)). Biological vulnerabilities, lack of female-led preventive measures, and lack of ability to negotiate condom use all contribute to global male-to-female transmission (R. Mallipeddi, LC Rohan,). Nanoparticle-based vaginal drug delivery systems for HIV prevention. Expert Opin Drug Deliv 7, 37-48 (2010); VM Ndesendo, V. Pillay, YE Choonara, E. Buchmann, DN Bayever, LC Meyer, A Review of current intravaginal drug delivery approaches employed for the prophylaxis of HIV / AIDS and prevention of sexually transmitted infections. AAPS PharmSciTech 9, 505-520 (2008)). Easy-to-administer, unobtrusive, and effective methods to protect women from vaginal HIV, HSV-2, and other viral infections may prevent a large number of infections worldwide. After failing to test 11 bactericides, CAPRISA004 was the first to demonstrate partial protection against HIV using a bactericidal agent (tenofovir) administered transvaginally in the form of a gel (tenofovir). QA Karim, SSA Karim, JA Frohlich, AC Grobler, C. Baxter, LE Mansoor, ABM Kharsany, S. Sibeko, KP Mlisana, Z. Omar, TN Gengiah, S. Maarschalk, N. Arulappan, M. Mlotshwa, L. Morris, D. Taylor, CT Grp, Effectiveness and Safety of Tenofovir Gel, an Antiretroviral Microbicide, for the Prevention of HIV Infection in Women. Science 329 , 1168-1174 (2010)). The difference between previous generation fungicides such as N9 and current generation fungicides is the site of action. Many current-generation bactericides, such as the nucleoside analogs tenofovir and acyclovir monophosphate, act intracellularly to inhibit viral replication, while previous generations direct pathogens in the vaginal lumen. Inactivated. However, some previous generation fungicides have been toxic to the vaginal epithelium, which has increased susceptibility to infection (OJ D'Cruz, FM). Uckun, Dawn of non-nucleoside inhibitor-based anti-HIV microbicides. J Antimicrob Chemother 57, 411-423 (2006)).</p><p>For maximally effective vaginal drug delivery, the locally delivered drug is evenly distributed, maintains a sufficiently high concentration, and has a folded vaginal epithelium (mucous folds) and cervix. It should stay in close proximity to the mucus. Several techniques, such as MRI (CK Mauck, D. Katz, EP Sandefer, MD Nasution, M. Henderson, GA Digenis, I. Su, R. Page, K. Barnhart, Vaginal distribution of Replens and KY Jelly using three imaging techniques. Contraception 77, 195-204 (2008)), Gamma-Scintigraphy (CK Mauck, D. Katz, EP Sandefer, MD Nasution, M. Henderson, GA Digenis, I. Su, R. Page, K. Barnhart, Vaginal distribution of Replens and KY Jelly using three imaging techniques. Contraception 77, 195-204 (2008); and BE Chatterton, S. Penglis, JC Kovacs, B. Presnell, B. Hunt, Retention and distribution of two 99mTc- DTPA labelled vaginal dosage forms. Int J Pharm 271, 137-143 (2004)), Vaginal Surgery (N. Poelvoorde, H. Verstraelen, R. Verhelst, B. Saerens, E. De Backer, GL dos Santos Santiago, C .Vervaet, M. Vaneechoutte, F. De Boeck, L. Van Bortel, M. Temmerman, JP Remon, In vivo evaluation of the vaginal distribution and retention of multi-particulate pellet formulation. Eur J Pharm Biopharm 73, 280-284 (2009)), and fiber optics (CK Mauck, D. Katz, EP Sandefer, MD Nasution, M. Henderson, GA Digenis, I. Su, R. Page, K. Barnhart, Vaginal distribution of Replens and KY Jelly using three imaging techniques. Contraception 77, 195-204 (2008)) was used to observe the distribution of gels and drugs following vaginal administration. These techniques are suitable for observing a rough distribution along the vaginal canal, but do not reveal invasion into the vaginal folds. Our study found that topical treatments may be well distributed along the length of the vaginal canal, but many of the collapsed epithelia remain untreated and unprotected. I proved that there was. Such untreated surfaces may contribute to the recent failure of some candidate fungicides for HIV in clinical trials (CW Hendrix, YJ Cao, EJ Fuchs, Topical microbicides to prevent HIV: clinical drug). development challenges. Annu Rev Pharmacol Toxicol 49, 349-375 (2009)). In addition, when a fluid or gel is administered to the vagina, it comes into direct contact with the rapidly flowing outer luminal mucus layer. Mucous sticky particles such as CP are trapped in this superficial mucous layer, thereby being removed from the mucous folds. In contrast, we have the ability of MPP to penetrate deeply into the mucous folds of mice and, when delivered hypotonically, provided complete epithelial coverage within just 10 minutes. ..</p><p>Diffusion of particles is not fast enough to result in such a uniform epithelial coating within minutes. Diffusions above about 100 μm require approximately several hours. However, the vaginal epithelium has great capacity for fluid absorption induced by osmotic gradients. Absorption of water in the mucus barrier assists the MPP in rapidly reaching the entire epithelial surface by advection, and then the loaded drug can be released for optimal uptake in the tissue. In contrast, water absorption is not beneficial to CP as it sticks and is captured and immobilized in the luminal mucus (Video 2).</p><p>Inadequate retention of therapeutically active compounds in the vaginal canal is another limiting factor for vaginal protection. For example, many vaginal spermicide provide protection for less than an hour (LJD Zaneveld, DP Waller, N. Ahmad, J. Quigg, J. Kaminski, A. Nikurs, C. De Jonge, Properties of a). new, long-lasting vaginal delivery system (LASRS) for contraceptive and antimicrobial agents. J. Androl 22, 481-490 (2001)). Other vaginal products were not sufficiently retained even after 6 hours (RF Omar, S. Trottier, G. Brousseau, A. Lamarre, G. Alexandre, MG. Bergeron, Distribution of a vaginal gel (Invisible Condom) before, during and after simulated sexual intercourse and its persistence when delivered by two different vaginal applicators: a magnetic resonance imaging study. Contraception 77, 47-455 (2008); N. Poelvoorde, H. Verstraelen, R. Verhelst, B. Saerens, E. De Backer, GL dos Santos Santiago, C. Vervaet, M. Vaneechoutte, F. De Boeck, L. Van Bortel, M. Temmerman, JP Remon, In vivo evaluation of the vaginal distribution and retention of a multi-particulate pellet formulation. Eur J Pharm Biopharm 73, 280-284 (2009); BE Chatterton, S. Penglis, JC Kovacs, B. Presnell, B. Hunt, Retention and distribution of two 99mTc-DTPA labeled vaginal dosage forms. Int J Pharm 271, 137-143 (2004)), requiring repeated doses for adequate protection. Similarly, CP does not penetrate deeply into the mucous layer, so more than 90% of it was flushed out of the vagina within 6 hours. In contrast, MPP provided enhanced delivery of the encapsulated model drug (FITC) for at least 24 hours compared to the soluble drug in the gel form. Therefore, MPP can provide a means for achieving strong once-daily topical vaginal administration, such as antiseptic for sexually transmitted diseases, for treatment.</p><p>Various "preliminary procedures" have been used to reduce the mucous barrier in conventional attempts to develop mucosal drug delivery systems for the vaginal canal. Y. Cu, CJ Booth, WM Saltzman, In vivo distribution of surface-modified PLGA nanoparticles following intravaginal delivery. J.Control Release, (10.1016 / j.jconrel. 2011.06.036); KA Woodrow, Y. Cu, CJ Booth, JK Saucier-Sawyer, MJ Wood, WM Saltaman, Intravaginal gene silencing using biodegradable polymer nanoparticles densely loaded with small-interfering RNA. Nat Mater 8, 526-533 (2009) ); T. Kanazawa, Y. Takashima, S. Hirayama, H. Okada, Effects of menstrual cycle on gene transfection through mouse vagina for DNA vaccine. Int J Pharm 360, 164-170 (2008); T. Kanazawa, Y. Takashima, Y. Shibata, M. Tsuchiya, T. Tamura, H. Okada, Effective vaginal DNA delivery with high transfection efficiency is a good system for induction of higher local vaginal immune responses. J Pharm Pharmacol 61, 1457-1463 2009)) KA Woodrow, Y. Cu, J. Booth, JK Saucier-Sawyer, MJ Wood, WM Saltaman, Intravaginal gene silencing using biodegradable polymer nanoparticles densely loaded with small-interfering RNA. Nat Mater 8, 526-533 (2009); AS Kask, X. Chen, JO Marshak, L. Dong, M. Saracino, D. Chen, C. Jarrahian, MA Kendall, DM Koelle, DNA vaccine delivery by densely-packed and short microprojection arrays to skin protects against vaginal HSV-2 challenge. Vacccine 28, 7483-7491 (2010)), or degrading enzyme (MM Seavey, TR) Mosmann, Estradiol-induced vaginal mucus inhibits antigen penetration and CD8 (+) T cell priming in response to intravaginal immunization. Vaccine 27,2342-2349 (2009)) is probably essential for the drug or gene delivery achieved in these studies. Is. In this case, it was found that pretreatment with lavage + swab wiping significantly improved the distribution of CP in the vagina and allowed the particles to coat the epithelium as well as MPP (Fig. 19). ). Pretreatments that remove the barrier are impractical for use in humans and may not be particularly suitable for fungicides intended to prevent sexually transmitted diseases. Healthy CVM itself is a slightly more effective barrier against viral infections (SK Lai, K. Hida, S. Shukair, YY Wang, A. Figueiredo, R. Cone, TJ Hope, L. Hanes, Human immunodeficiency virus type 1 is trapped by acidic but not by neutralized human cervicovaginal mucus. J Virol 83, 11196-11200 (2009)). Effective epithelial coverage can be achieved by using MPP without the need to degrade or remove the mucous barrier.</p><p>PEG coatings have been widely used in the development of polymeric drug carriers that are not easily recognized by the immune system (BC Tang, M. Dawson, SK Lai, YY Wang, JS Suk, M. Yang, P. Zeitlin, MP Boyle, J. Fu, J. Hanes, Biodegradable polymer nanoparticles that rapidly penetrates the human mucus barrier. Proc Natl Acad Sci USA 106, 19268-19273 (2009)). The dense PEG coating has been demonstrated to rapidly penetrate mucus without causing inflammation in the vaginal canal of mice. In contrast, administration of uncoated CP evoked an acute inflammatory response similar to administration of N9. Moreover, the cytokine levels associated with daily administration of MPP were indistinguishable from HEC placebo gels. Elevated IL-1α and IL-1β levels associated with epithelial damage occurred after daily administration with both N9 and TFV vehicle gels. A tenofovir-containing version of this gel has been shown to have complete protection against HIV in an in vitro transplant model, with complete protection occurring despite visible epithelial shedding (LC Rohan, BJ Moncla,). RP Kunjara Na Ayudhya, M. Cost, Y. Huang, F. Gai, N. Billitto, JD Lynam, K. Pryke, P. Graebing, H. Hopkins, JF Rooney, D. Friend, CS Dezzutti, In vitro and ex vivo testing of tenofovir shows it is effective as an HIV-1 microbicide. PLoS One 5, e9310 (2010)). Previous studies suggest that glycerol in TFV gels may be responsible for the toxicity observed in mice (TR Moench, RJ Mumper, TE Hoen, M. Sun, RA Cone, BMC Infect Dis 10, 331 (2010)). Microbicide investigateds can greatly increase susceptibility to genital herpes transmission in the mouse.</p><p>Although the mouse is a useful animal model for developing vaginal products, there are important differences in vaginal physiology between mice and humans. First, the estrus cycle occurs in a cycle spanning 4-5 days, in contrast to the 28-day human menstrual cycle. Changes in the human vaginal epithelium are relatively minimal during the menstrual cycle, whereas epithelial shedding is followed by substantial proliferation during the four stages of the mouse estrus cycle (BG Smith, EK Brunner). , The structure of the human vaginal mucosa in relation to the menstrual cycle and to pregnancy. American Journal of Anatomy 54, 27-85 (1934); AK Ildgruben, IM Sjoberg, M.-LKC Hammarstrom, Influence of hormonal contraceptives on the immune cells and thickness of human vaginal epithelium. Obstetrics &amp; Gynecology 102, 571-582 (2003)). The end of estrus and the beginning of estrus in mice are most similar to those of human vaginal epithelium (BG Smith, EK Brunner, The structure of the human vaginal mucosa in relation to the menstrual cycle and to pregnancy). .American Journal of Anatomy 54, 27-85 (1934); AW Asscher, CH De Boer, CJ Turner, Cornification of the human vaginal epithelium. J.Anat 90, 547-552 (1956)). At these stages, there is significant bacterial colonization, including the peak presence of Lactobacillus (HM Cowley, GS Heiss, Changes in Vaginal Bacterial Flora During the Oestrous Cycle of the Mouse. Microbial Ecology in Health and Disease 4, 229- 235 (1991)). In addition, the effects of estradiol are both permeable to MPP and eliminated in approximately hours, causing active secretion of mucus found in mice to resemble humans (HM Cowley, GS). Heiss, Changes in Vaginal Bacterial Flora During the Oestrous Cycle of the Mouse. Microbial Ecology in Health and Disease 4,229-235 (1991); LB Corbeil, A. Chatterjee, L. Foresman, JA Westfall, Ultrastructure of cyclic changes in the murine uterus, cervix, and vagina. Tissue Cell 17, 53-68 (1985); CG Rosa, JT Velardo, Histochemical localization of vaginal oxidative enzymes and mucins in rats treated with estradiol and progesterone. Ann NY Acad Sci 83, 122-144 (1959)). Therefore, the IE mouse model is considered to be a valuable model in addition to the commonly used DP model for investigating vaginal delivery methods. Estradiol can be used to synchronize mice in estrus, but does not "hold" them in estrus. Those mice continue the cycle, while DP treatment allows the mice to remain in estrus for days to weeks (C. Kaushic, AA Ashkar, LA Reid, KL Rosenthal, Progesterone increases susceptibility and decreases immune responses to genital herpes infection. J Virol 77, 4558-4565 (2003)).</p><p>The ability of MPP to rapidly penetrate human cervical vagina and mouse vaginal mucus, and MPP significantly in terms of coating speed and uniformity and retention time compared to conventional mucous sticky nanoparticles. It was shown to improve. CP elicited an acute inflammatory response similar to known irritant N9, but MPP, like placebo gels, did not elicit a detected inflammatory response. Vaginal administration of MPPs loaded with acyclovir monophosphate was more effective in protecting mice from vaginal HSV-2 infection than soluble drugs, even at concentrations of soluble drugs greater than 10-fold. .. These results motivate the further development of MPPs for the treatment of sexually transmitted diseases, contraception, and other cervical disorders to deliver safe and effective drugs to the vagina.</p><p>(Example 8) This non-limiting example is the relationship between the relative velocity of polystyrene (PS) particles coated with Pluronic® F127 in mucus and the density of Pluronic® F127 on the particle surface. Is shown.</p><p>In a set of experiments, an aqueous dispersion of carboxylated PS nanoparticles (200 nm, 0.5% w / v) was equilibrated in the presence of various concentrations of Pluronic® F127 at room temperature for at least 24 hours. .. The density on the surface of the obtained PS / Pluronic® F127 nanoparticles of the Pluronic® F127 molecule was quantified as follows. Particle settling was completed by ultracentrifugating the PS / Pluronic® F127 mixture. As a result, PS-bound Pluronic® F127 precipitated with the particles; PS-unbound Pluronic® F127 remained in the supernatant. Obtained supernatant (C_{F127, free}The concentration of Pluronic® F127 in) was measured by gel permeation chromatography (GPC). This experiment utilized an Agilent 1100 HPLC system equipped with a G1362A refractive index detector and an Agilent PLgel 5 μm Mixed-C column for analysis. Combined Pluronic® F127 (C)_{F127, join}) Concentration was calculated as follows: C_{F127, join}= C_F127-C_{F127, free}(In the formula, C_F127Is the total concentration of Pluronic® F127 present in the mixture. Then PS surface area (F127 / nm)²The number of Pluronic® F127 molecules per) was calculated as follows:<math num="2"><img file="JP2022017589A_D0017.tif" /></math>(In the formula, N_AIs the Avogadro number, C_{F127, join}Is the molar concentration (molar / L) of the bound Pluronic® F127 and SA is the specific surface area (nm) of the PS particles calculated from the manufacturer's (Invitrogen) specifications.²/ g) and C_PSIs the mass concentration (g / L) of PS in the mixture. The number average molecular weight of Pluronic® F127 specified by the manufacturer (BASF) was used in the calculation.</p><p>Using fluorescence microscopy and multiple particle tracking software as described in Examples 1 and 4-6, the mucus penetration capacity of PS / Pluronic® F127 particles was relative to human cervical mucus. Measured as velocity. In particular, the sample is the particles of interest, the negative control is 200 nm fluorescent carboxylated-modified polystyrene particles without a polymer coating, and the positive control is a high density coating of 2 or 5 kDa PEG. It was a 200 nm fluorescent polystyrene particle with (with a well-established, lower mucosal adhesion behavior). Samples, negative controls, and positive controls were distinguished from each other by their fluorescent color.</p><p>Relative velocity of particles in cervical mucus was characterized using fluorescence microscopy and multi-particle tracking software as described in Examples 1 and 4-6. In a typical experiment, a 0.5 μL particle suspension was added to 20 μL of fresh cervical mucus with positive and negative controls. Using a fluorescent microscope equipped with a CCD camera, at a temporary resolution of 66.7 ms (15 frames / sec), at a magnification of 100 ×, each type of particle: sample, negative control, and positive control each. A 15-second video was recorded from several areas in the sample. Then, using state-of-the-art image processing software, individual trajectories of multiple particles were measured over a timescale of at least 3.335 seconds (50 frames).</p><p>The results shown in Figure 25 show that the relative velocities of PS particles coated with Pluronic® F127 in mucus increased as the density of Pluronic® F127 molecules on the particle surface increased. Indicated.</p><p>Other Embodiments Although some embodiments of the invention have been described and exemplified herein, those skilled in the art can perform functions and / or results and / or the present specification. Various other means and / or structures for obtaining one or more of the advantages described in are readily envisioned, and each such modification and / or modification is within the scope of the invention. It is considered to be. More generally, one of ordinary skill in the art intends that all parameters, dimensions, substances, and configurations described herein are exemplary and actual parameters, dimensions, substances, and /. Alternatively, it will be readily appreciated that the configuration will depend on the particular application (s) in which the teachings of the invention are used. One of ordinary skill in the art can recognize or ascertain many equivalents for a particular embodiment of the invention described herein using only conventional testing. Accordingly, the embodiments listed above are presented by way of example only and are specifically described and within the scope of the appended claims and their equivalents, in a manner other than the claimed invention. It shall be understood that The present invention is directed to the individual features, systems, articles, substances, kits, and / or methods described herein, respectively. Moreover, any combination of two or more such features, systems, articles, substances, kits, and / or methods are consistent with such features, systems, articles, substances, kits, and / or methods. If so, it is included within the scope of the present invention.</p><p>The indefinite articles "a" and "an", as used herein and in the claims, are understood to mean "at least one" unless explicitly indicated to be the opposite. Should be.</p><p>The phrase "and / or", as used herein and in the claims, means that "either or both" of the elements are thus combined, i.e., if any. Is an element that exists in a connected manner, and in other cases it should be understood that it exists in a non-connected manner. Elements specifically identified by the "and / or" clause, whether or not they are associated with such elements as specifically identified, unless explicitly indicated to be the opposite. Other elements may be present. Thus, as a non-limiting example, the reference to "A and / or B" when used in connection with an unrestricted language such as "contains", in one embodiment, does not include A (other than B). (May contain elements of); in another embodiment B without A (may contain elements other than A); in yet another embodiment both A and B (may contain other elements). ) Etc. can be pointed out.</p><p>As used herein in the specification and claims, "or" should be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" shall be interpreted as including all, i.e. include at least one of the numbers or of the list of elements. Not only does it contain more than one item, but it may also contain more unlisted items. The only term that clearly indicates the opposite, for example, "only one of" or "exactly one of", or "consisting of" when used in the claims, is a number. Or it would refer to the inclusion of exactly one element in the list of elements. In general, the term "or" as used herein is an exclusive term, eg, "any", "one of", "only one of" or "exactly". When preceded by "one" or the like, it shall be construed to indicate an exclusive alternative form (ie, "one but not both"). When "becomes basic" is used within the claims, it should have its usual meaning as used in the field of patent law.</p><p>As used herein and in the claims, the phrase "at least one" does not necessarily include at least one of each and every element specifically listed in the list of elements. Instead of excluding any combination of elements in the list of elements, from any one or more of the elements in the list of elements in relation to the list of one or more elements. It should be understood to mean at least one element selected. This definition also refers to the elements specifically identified in the list of elements pointed to by the phrase "at least one", whether or not they are related to these specifically identified elements. Allows elements other than may be present. Thus, as a non-limiting example, "at least one of A and B" (or equivalently, "at least one of A or B" or equivalently "at least one of A and / or B". One ") can, in one embodiment, refer to at least one A (which may contain more than one A) without the presence of B (may contain elements other than B); In another embodiment, A is absent and can refer to at least one B (which may contain more than one B) (may contain elements other than A); yet another embodiment. Can refer to at least one A (which may contain more than one A) and at least one B (which may contain more than one B) (which may contain other elements).</p><p>In the claims, and in the above specification, all the connecting words, for example, "comprising", "including", "holding", "having", "containing", "including (comprising)". It should be understood that "engaging", "holding", etc. mean that there are no restrictions, that is, they include, but are not limited to. The only fillers "consisting of" and "consisting of essentially" shall be closed or semi-closed, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03. That's right.</p><p>[Additional Notes]</p><p>[Appendix 1]</p><p>A composition comprising a plurality of coated particles, wherein the coated particles are core particles containing a solid drug or a salt thereof, and the drug or a salt thereof is anywhere in the pH range. A coating containing a core particle; and a surface modifier surrounding the core particle, having a water solubility of about 1 mg / mL or less at 25 ° C, wherein the drug or salt thereof constitutes at least about 80% by weight of the core particle. The surface modifier comprises a triblock copolymer comprising a hydrophilic block-hydrophobic block-hydrophilic block arrangement, wherein the hydrophobic block has a molecular weight of at least about 2 kDa and the hydrophilic block is Consists of at least about 15% by weight of the triblock copolymer, the hydrophobic block associates with the surface of the core particles, the hydrophilic block is present on the surface of the coated particles, and the coated particles are present. Hydrophilic and the surface modifier is at least about 0.001 molecule / nm on the surface of the core particles.²A composition comprising a coating, which is present at a density of, wherein the coated particles have a relative velocity greater than 0.5 in mucus.</p><p>[Appendix 2]</p><p>A method comprising delivering a composition comprising a plurality of coated particles to a mucous membrane, wherein the coated particles are core particles comprising a solid drug or a salt thereof, wherein the drug or a salt thereof. Has a water solubility of about 1 mg / mL or less at 25 ° C at any point in the pH range, and the drug or salt thereof constitutes at least about 80% by weight of the core particles; and core particles. A coating comprising a surface modifier that surrounds the surface modifier comprising a triblock copolymer comprising a hydrophilic block-hydrophobic block-hydrophilic block arrangement, wherein the hydrophobic block has a molecular weight of at least about 2 kDa. The hydrophilic block comprises at least about 15% by weight of the triblock copolymer, the hydrophobic block associates with the surface of the core particles and the hydrophilic block is present on the surface of the coated particles. The coated particles are made hydrophilic so that the surface modifier is at least about 0.001 molecule / nm on the surface of the core particles.²A method comprising a coating, which is present at a density of, wherein the coated particles have a relative velocity greater than 0.5 in mucus.</p><p>[Appendix 3]</p><p>A method of forming coated particles, in which the core particles are combined with a solution containing a surface modifier, wherein the core particles have a solubility of about 1 mg / mL or less in a solution at 25 ° C. And contains solid pharmaceuticals or salts thereof constituting at least about 80% by weight of each of the core particles; a step of coating the core particles with a surface modifier to form coated particles, wherein the surface modification. The agent comprises a triblock copolymer comprising a hydrophilic block-hydrophobic block-hydrophilic block arrangement, wherein the hydrophobic block has a molecular weight of at least about 2 kDa and the hydrophilic block is at least about 15 of the triblock copolymer. Consists of% by weight, the hydrophobic block associates with the surface of the core particles, the hydrophilic block is present on the surface of the coated particles, making the coated particles hydrophilic and the coated particles , Has a relative velocity greater than 0.5 in mucus; including methods.</p><p>[Appendix 4]</p><p>The composition according to Appendix 1, wherein the surface modifier is covalently bonded to the core particles.</p><p>[Appendix 5]</p><p>The composition according to Appendix 1, wherein the surface modifier is adsorbed on the core particles in a non-covalent bond.</p><p>[Appendix 6]</p><p>The composition according to Appendix 1, wherein the surface modifier is present on the surface of the coated particles at a density of at least about 0.01 molecules per square nanometer.</p><p>[Appendix 7]</p><p>The composition according to Appendix 1, wherein the hydrophilic blocks of the triblock copolymer make up at least about 30% by weight of the triblock copolymer.</p><p>[Appendix 8]</p><p>The composition according to Annex 7, wherein the hydrophobic block of the triblock copolymer has a molecular weight of at least about 3 kDa.</p><p>[Appendix 9]</p><p>The composition according to Appendix 8, wherein the triblock copolymer is poly (ethylene oxide) -poly (propylene oxide) -poly (ethylene oxide) or poly (ethylene glycol) -poly (propylene oxide) -poly (ethylene glycol).</p><p>[Appendix 10]</p><p>The composition according to Annex 7, wherein the hydrophilic block of the triblock copolymer comprises poly (ethylene oxide) or poly (ethylene glycol), or a derivative thereof.</p><p>[Appendix 11]</p><p>The composition according to Annex 10, wherein the poly (ethylene oxide) or poly (ethylene glycol) block has a molecular weight of at least about 2 kDa.</p><p>[Appendix 12]</p><p>The composition according to Appendix 1, wherein the hydrophobic block of the triblock copolymer is poly (propylene oxide).</p><p>[Appendix 13]</p><p>The composition according to Annex 12, wherein the poly (propylene oxide) block has a molecular weight of at least about 3 kDa.</p><p>[Appendix 14]</p><p>The composition according to Appendix 1, wherein the surface modifier is present in the solution at a concentration of at least about 0.1 (weight / volume)%.</p><p>[Appendix 15]</p><p>The composition according to Appendix 1, wherein each of the core particles comprises a crystalline drug or a salt thereof.</p><p>[Appendix 16]</p><p>The composition according to Appendix 1, wherein each of the core particles comprises an amorphous drug or a salt thereof.</p><p>[Appendix 17]</p><p>The composition according to Appendix 1, wherein each of the core particles contains a salt of a solid pharmaceutical product.</p><p>[Appendix 18]</p><p>The composition according to Appendix 1, wherein the pharmaceutical product is at least one of a therapeutic agent or a diagnostic agent.</p><p>[Appendix 19]</p><p>The composition according to Annex 1, wherein the pharmaceutical product is at least one of a small molecule, a peptide, a peptide mimetic, a protein, a nucleic acid, or a lipid.</p><p>[Appendix 20]</p><p>The composition according to Appendix 1, wherein the drug or a salt thereof has a water solubility of about 0.1 mg / mL or less at 25 ° C.</p><p>[Appendix 21]</p><p>The composition according to Appendix 1, wherein the pharmaceutical product comprises at least about 85% by weight of the core particles.</p><p>[Appendix 22]</p><p>The composition according to Appendix 1, wherein the core particles have an average size of at least about 20 nm and about 1 μm or less.</p><p>[Appendix 23]</p><p>The composition according to Appendix 1, wherein the coated particles have an average size of at least about 20 nm and about 1 μm or less.</p><p>[Appendix 24]</p><p>The composition according to Appendix 1, wherein the coated particles diffuse in human cervical mucus on a time scale of 1 second with a diffusivity greater than 1/500 of the diffusivity at which the particles diffuse in water.</p><p>[Appendix 25]</p><p>The composition according to Appendix 1, wherein the coated particles have a relative velocity greater than 0.8 in mucus.</p><p>[Appendix 26]</p><p>The composition according to Appendix 1, wherein the mucus is human cervical mucus.</p><p>[Appendix 27]</p><p>A pharmaceutical composition comprising the composition according to Annex 1 and one or more pharmaceutically acceptable carriers, additives, and / or diluents.</p><p>[Appendix 28]</p><p>A pharmaceutical preparation suitable for inhalation, injection, or topical administration into a mucous membrane, which comprises the composition according to Appendix 1.</p>