JP2007507489A - Water-soluble nanoparticle-encapsulated composite - Google Patents

Abstract

  A nano-dispersion of water-soluble and stable nano-sized particles is provided as a constituent of a hydrophilic encapsulated complex of the active compound in an amorphous state surrounded and encapsulated by an amphiphilic polymer. The encapsulated complex is stabilized by a worthless interaction between the active compound and the polymer. The following pairs of active compounds / amphiphilic polymers are provided: (i) azithromycin / polysaccharide or polyvinyl alcohol, or clarithromycin / alginic acid or chitosan; (ii) donepezil hydrochloride / polysaccharide; (iii) azole compound / Polysaccharides, polyacrylic acid, copolymers of polyacrylic acid, polymethacrylic acid, and copolymers of polymethacrylic acid; and (iv) taxane / gelatin.

Description

  The present invention belongs to the field of nanoparticles. More particularly, the present invention is water soluble and stable, composed of an encapsulated complex of active compounds such as pharmaceuticals or agrochemicals surrounded by a suitable amphiphilic polymer and contained therein. Related to nano-dispersion of various nano-sized particles and to the method for realizing nano-dispersion described above.

  Two troublesome obstacles to effective drug delivery and hence treatment of illness are solubility and stability. In order to be absorbed by the human body, a compound must be soluble in both water and fat (lipid). However, solubility in water is often associated with low fat solubility and vice versa.

  More than one-third of drugs registered in the US Pharmacopeia and about 50% of new chemicals (NCEs) are insoluble or poorly insoluble in water. More than 40% of drug molecules and drug compounds are insoluble in the human body. Despite this, lipophilic drug substances with low water solubility are an increasing class of drugs with increasing applications in various therapeutic fields and for various pathologies. Today, over 2500 macromolecules are in various stages of development, and over 5500 small molecules are under development (see Drug Delivery Companies Report 2001, p.2, www.pharmaventures.com). Each of these current companies that focus on these large and small molecules has their own regulations and restrictions on both the large and small molecules of interest.

  Solubility and stability issues are a major prescription disorder that hinders the development of therapeutics. Water solubility is an essential but often elusive property for the formulation of complex organic structures found in medicine. Traditional formulation systems for highly insoluble drugs required a combination of organic solvents, surfactants, and extreme pH conditions. These prescriptions are often frustrating for the patient and may lead to adverse reactions. Sometimes these methods are inadequate for dissolving a sufficient amount of drug for parenteral formulation. In such cases, the physician may take an “excessive” dose, such as for a poorly soluble vitamin. In most cases, this overdose does not cause harm because the unabsorbed amount is excreted from the body with the urine. However, overdosing is a waste of large amounts of active substance.

  The size of the drug molecule also plays a major role in bioavailability as well as its solubility and stability. Bioavailability refers to the degree to which a drug becomes available to a target tissue or any other in vivo target (ie, receptor, cancer cell, etc.) after it is administered into the body. Poor bioavailability is a significant problem faced in the development of pharmaceutical compositions, especially when the active ingredients contained are poorly soluble in water. Drugs with poor water solubility tend to be excreted from the gastrointestinal tract before being absorbed into the circulatory system. It is known that the efficiency of dissolution of a particulate drug can increase with increasing surface area, ie, decreasing particle size.

  In recent years, interest in nanotechnology, which is an operation at the nanoscale, has increased rapidly. Nanotechnology is not a whole new field: colloidal sols and supported platinum catalysts are nanoparticles. Nevertheless, recent interest in the nanoscale has produced materials used for and in drug delivery, among many others. Nanoparticles are generally considered solids whose particle size varies between 1 and 1000 nm.

  There are a number of solubilization techniques, such as liposomes, cyclodextrins, microencapsulation, and dendrimers, but each of these techniques has a number of significant drawbacks.

  Phospholipids exposed to an aqueous environment form a bilayer structure called liposomes. Liposomes are fine spherical structures composed of phospholipids that were first discovered in the early 1960s. In aqueous media, phospholipid molecules are amphiphilic and spontaneously organize into self-closing bilayers as a result of hydrophilic and hydrophobic interactions. The resulting vesicles, called liposomes, therefore have the property of becoming a potential carrier for biologically active hydrophilic molecules and drugs in vivo when they are suspended. Thus, it is wrapped like a capsule in the inner part of the aqueous medium. Lipophilic substances may also be transported embedded in the liposome membrane. Liposomes resemble biological membranes and have been used for many years as a tool for solubilizing bioactive molecules that are insoluble in water. They are non-toxic, biodegradable and can be used for specific target organs.

  Liposome technology allows the preparation of smaller to larger vesicles using unilamellar vesicles (ULV) and multilamellar vesicles (MLV). MLV is produced by mechanical stirring. Large ULVs are prepared from MLVs by extruding under pressure through a membrane of known pore size. Its size is 200 nm or less in diameter; however, liposomes can be custom designed for most needs by changing lipid content, changing surface, and preparation methods.

  As a drug carrier, liposomes offer several possibilities, including the ability to carry significant amounts of drug, the relative ease of preparation, and low toxicity when natural lipids are used. Has certain advantages. However, there are common problems faced by liposomes: low stability, short shelf life, poor tissue specificity, and toxicity when used with non-natural lipids. In addition, uptake by phagocytes reduces circulation time. Furthermore, preparing liposome formulations that exhibit a narrow size distribution is both a costly and challenging task under demanding conditions. Also, membrane clogging often occurs during the high volume production required to produce a particular drug as a pharmaceutical.

  Cyclodextrins are crystalline, water-soluble, cyclic, non-reducing oligosaccharides composed of 6, 7 or 8 glucopyranose units, referred to as alpha, beta, and gamma cyclodextrins, respectively, to form inclusion complexes It has long been known as a product with capacity. The cyclodextrin structure gives a molecule shaped like a hollow cone with an outer hydrophilic surface and an inner hydrophobic cavity.

  The hydrophilic surface creates good water solubility for cyclodextrins, and the hydrophobic cavity provides a suitable environment for surrounding, enclosing, or enclosing drug molecules. This linkage may isolate the drug from the aqueous solvent and improve the drug's water solubility and stability. For a long time, most cyclodextrins have only been of scientific interest due to their limited availability and high price.

  As a result of in-depth research and advances in enzyme technology, cyclodextrins and their chemically modified derivatives are now commercially available, creating new technologies, namely molecular level packaging. However, cyclodextrins are inconvenient. An ideal cyclodextrin will show both oral and systemic safety. It will have water solubility over the original cyclodextrin and will retain or surpass its complex formation properties. However, the disadvantages of cyclodextrins are the limited available space for the active molecule to be centered inside, the lack of pure stability of the complex, and limited market availability. And being expensive.

  Microencapsulation means that small packets of active composition in gas, liquid, or solid (also referred to herein as “core material” and used interchangeably with “core material”) are active from the surrounding environment. A process by being packaged in a second material for the purpose of protecting the composition. These capsules range in size from 1 micron (1/1000 of a millimeter) to approximately 7 millimeters and release their contents at a later time by a method suitable for the application.

  There are four typical mechanisms by which the core material is released from the microcapsule. (1) mechanical rupture of the capsule wall, (2) decomposition of the wall, (3) dissolution of the wall, and (4) diffusion through the wall. Less common release mechanisms include removal (slow erosion of the shell) and biodegradation.

  Microencapsulation covers a range of techniques in which a material is coated to obtain a micropackaging of the active compound. Coating is done for many other purposes, such as to stabilize the material, to confine the flavor, to prepare a material that flows freely from other clogging substances, and so on. This technology has been successfully applied to the food additive industry and agriculture. However, requiring a relatively high production cost for many of the formulations is a significant drawback.

  In the case of nanoencapsulation and nanoparticles (which are preferably spherically shaped and hence nanospheres), two types of systems with different internal structures are possible. (I) a matrix type system composed of entanglements of oligomer or polymer units, defined as nanoparticles or nanospheres, and (ii) an oily center surrounded by a polymer wall, defined as nanocapsules A reservoir-type system composed of

  Depending on the nature of the material used to prepare the nanosphere, there are the following classifications: (A) an amphiphilic polymer that undergoes a crosslinking reaction during nanosphere preparation; (b) a monomer that polymerizes during nanoparticle preparation; and (c) first dissolved in an organic solvent; Then a hydrophobic polymer, precipitated under controlled conditions to produce nanoparticles.

  There are problems associated with the use of polymers in micro and nano encapsulation. Use of toxic emulsifiers in emulsions or dispersions, application of high shear forces during polymerization or emulsification, poor biocompatibility and biodegradability, hydrophilic and hydrophobic parts Balance, etc. These characteristics cause inadequate drug release.

  Dendrimers are a class of polymers classified by their highly branched, tree-like structure. They are synthesized in an iterative manner from ABn monomers, with each iteration of adding layers, or “generations” of the growing polymer. Ten generations or more of dendrimers have been synthesized with molecular weights exceeding 106 kDa. One important feature of dendrimer polymers is their narrow molecular weight distribution. Indeed, depending on the synthetic strategy used, dendrimers with molecular weights above 20 kDa can be synthesized as a single compound.

  Dendrimers, like liposomes, exhibit encapsulation properties and can confine molecules in the interior space. Because they are single molecules and not aggregates, drug-dendrimer complexes are expected to be much more stable than liposome-drug complexes. Dendrimers are therefore considered one of the most promising delivery vehicles for drug delivery systems. However, dendrimer technology is still in the research stage, and it is estimated that it will take several years before it is industrially applied as a safe and effective drug delivery system.

  What is needed is a safe, biocompatible, stable and effective composition of nano-sized particles of an active composition to improve bioavailability, overcoming the problems inherent in past inventions Drug delivery system.

  U.S. Published Patent Publication No. US 2003/0129239, assigned to the present applicant, discloses general techniques for the preparation of nanoparticles described in the present invention.

  Lipophilic and hydrophilic compounds solubilized in the form of nano-sized particles, or “nanoparticles”, are used in pharmacology, food additive production, cosmetics, and agriculture, as well as pet food, veterinary products, And it can be used in common with other usages.

  The present invention provides a nanodispersion of nanoparticles and a method for producing soluble nanoparticles, and in particular, a method for producing encapsulated composites of water-insoluble lipophilic organic materials and water-soluble hydrophilic organic materials. To do.

  An inclusion complex, by definition, is a complex in which one component called a “host” forms a cavity in which a molecular entity of a second chemical species called a “guest” is located. Thus, according to the present invention, the soluble nanoparticles are immobilized in a cavity or space in which the host is an amphiphilic polymer or group of polymers and the guest is formed by the aforementioned polymer host, or It can be defined as constituting an encapsulated complex that is a protected, active complex molecule.

  In accordance with the present invention, the inclusion complex comprises an active complex molecule that interacts with the polymer by a non-valent interaction to form the polymer active compound as a separate molecular entity. An obvious advantage and unique feature of the inclusion complex of the present invention is that no new chemical bonds are formed and any existing bonds are not destroyed during the formation of the inclusion complex. The particles making up the encapsulated complex are nano-sized in size and do not undergo any change in the drug molecule itself when packaged or advantageously wrapped by a polymer.

  The outer surface of the encapsulation complex consists of a polymer that, if the active compound is a drug molecule, carries it to the target destination. Depending on the polymer used to form the nanoparticles, the drugs and medicines in the complex can easily and quickly reach specific parts of the body. The selected polymer and active compound will also provide nanoparticles capable of multi-, multi-stage and / or controlled release of drugs or pharmaceuticals in the body.

  The nanoparticles of this invention are stable for a long period of time, may be manufactured at low cost, and may improve the overall bioavailability of the active compound.

  In particular, the present invention relates to nano-particles of water-soluble and stable nano-sized particles that constitute a hydrophilic inclusion complex consisting essentially of an active compound surrounded by an amphiphilic polymer. A dispersion wherein the active compound in the amphiphilic polymer is in an amorphous state and the encapsulated complex is stabilized by an unvalent interaction between the active compound and the surrounding amphiphilic polymer And the encapsulation complex is provided with a nanodispersion selected from the group consisting of:

  (I) an encapsulation complex in which the active compound is clarithromycin and the amphiphilic polymer is alginic acid or chitosan, or the active compound is azithromycin and the amphiphilic polymer is a polysaccharide or polyvinyl alcohol;

  (Ii) an encapsulated complex wherein the active compound is donepezil hydrochloride and the amphiphilic polymer is a polysaccharide;

  (Iii) the group wherein the active compound is an azole antifungal agent and the amphiphilic polymer is a polysaccharide, polyacrylic acid, polyacrylic acid copolymer, polymethacrylic acid, and polymethacrylic acid copolymer An inclusion complex selected from; and

  (Iv) An encapsulated complex in which the active compound is a taxane and the amphiphilic polymer is gelatin.

  The present invention also provides a pharmaceutical composition that constitutes a stable nanodispersion in this invention.

  The nanoparticles of the present invention constitute an insoluble or soluble active compound or core encased in a water soluble amphiphilic polymer. Different polymer types can be used according to the invention for any selected active compound that is lipophilic or hydrophilic. A polymer, or group of polymers, takes into account various physical properties of both the lipophilic or hydrophilic active compound and the resulting interaction of the compound within the active compound / polymer nanoparticles. Selected by different algorithms. This technique is fully described in US 2003/0129239 referenced above.

  One important parameter in the selection of a polymer or group of polymers is the measurement of the molecular balance of the HLB, the hydrophilic and lipophilic part of the compound. Within the HLB international scale from 0 to 20, lipophilic molecules are HLB less than 6 and hydrophilic molecules are HLB greater than 6. Thus, according to the present invention, the polymer HLB is selected in such a way that after conjugation with the active compound, the complex is water soluble if the overall resulting HLB value of the complex is greater than 8. Is done.

  As used herein, the term “non-crystalline” refers to materials that are both amorphous or disordered crystalline. In the preferred embodiment, the material is amorphous. It is known to those skilled in the art that an amorphous state is preferred for drug delivery as it may certainly increase bioavailability.

  As used herein, the terms “water-soluble nanoparticles”, “aqueous solution of nanoparticles” and “nanodisperse” are used interchangeably and are both meant to refer to the same thing, ie It is intended to refer to a fine dispersion of nanoparticles that has the appearance of a solution but is not a traditional aqueous solution.

  As used herein, the terms “stable nanodispersion” and “nanodispersion of water-soluble and stable nanosize particles” are used interchangeably and are both interchangeable, ie, stable and fine nanoparticles. It is intended to mention dispersion.

  The stability of nanoparticles and encapsulated composites does not only have one meaning. While the nanoparticles remain in the dispersion medium, they will be stable over time as part of the nanocomposite. Nanodispersion is stable for a long time without step separation. Furthermore, the irregular state will also be maintained for a long time.

  It is noteworthy that in the process used in the present invention, the system components do not result in micelles and do not form a classical distributed system. The technology of the present invention provides the following:

  (I) After forming the encapsulated complex, the active compound that slightly dissolves or does not dissolve (or even cannot be wetted) becomes pseudo-soluble. When the particle size is about 20 to 30 nm, the resulting material becomes soluble and the appearance is transparent rather than opaque;

  (Ii) After the dispersion of the active compound to the nanosize and fixation by the polymer forming the encapsulated complex, in vivo, similar to improved revision, improved absorption, and improved biological activity as in physiological fluids In addition, transmission to a stable amorphous, preferably amorphous state is achieved;

  (Iii) Biologically active compound crystals become amorphous and thus exhibit improved biological activity.

  In the most preferred embodiment of the invention, if the size deviation is not greater than 20% and the particle size is within the nano range, i.e. less than 1000 nm, more than 80% of the nanoparticles in the nanodispersion are within the size range.

  In an advantageous and preferred embodiment of the invention, the polymer molecules in the polymer solution “wrap” the active compound via a non-valent interaction. The term “valueless” as used herein is a non-covalent, non-ionic, and non-semipolar bond and / or interaction, eg, between a polymer and an active compound in an encapsulated complex. Includes forces such as electrostatic forces, van der Waals forces, and hydrogen bonds, which result in a non-valent interaction of anchoring the active compound in the polymer, resulting in reduced molecular flexibility of the active compound and polymer. . The formation of any valuable bond will change the characteristics or properties of the active compound. The formation of non-valent bonds retains the structure and properties of lipophilic compounds, which is particularly important when the active compound is a pharmaceutical.

  The present invention provides a nanodispersion of water-soluble and stable nanosized particles comprising a hydrophilic inclusion complex consisting essentially of an active compound surrounded by an amphiphilic polymer. provide. Here, the active compound in the aforementioned amphiphilic polymer is in an amorphous state, and the aforementioned encapsulated complex is formed by a non-valent interaction between the active compound and the surrounding amphiphilic polymer. Stabilized and the aforementioned inclusion complex is selected from the group consisting of: That is,

  (I) when the active compound is a macrolide antibiotic selected from clarithromycin and azithromycin, the active compound is clarithromycin, the amphiphilic polymer is alginic acid or chitosan, and the active compound is azithromycin An encapsulated complex wherein the functional polymer is a polysaccharide or polyvinyl alcohol;

  (Ii) an encapsulated complex wherein the active compound is donepezil hydrochloride and the amphiphilic polymer is a polysaccharide;

  (Iii) the active compound is an azole compound, and the amphiphilic polymer is selected from the group consisting of polysaccharides, polyacrylic acid, copolymers of polyacrylic acid, polymethacrylic acid, and copolymers of polymethacrylic acid Encapsulated complex; and

  (Iv) An encapsulated complex in which the active compound is a taxane and the amphiphilic polymer is gelatin.

  In one preferred embodiment, the nanoparticles of this invention constitute an inclusion complex where the active compound is clarithromycin, a macrolide antibiotic, or azithromycin, a first azalide antibiotic. These macrolides are large, lipophilic molecules, have a broad range of antibiotic activity against a wide variety of bacteria, and can be used as both human and veterinary medicines. Macrolide antibiotics are particularly useful for the treatment of respiratory infections.

  Suitable polymers for the preparation of inclusion complexes with macrolide antibiotics are natural or modified polysaccharides. In one embodiment, the polysaccharide is a starch that is desirably mostly linear, i.e., that of an starch in which anhydroglucose units form a linear chain (-D-1,4 glucosidic linkages). A starch that is rich in amylose as a component and low in amylopectin as a component of a polymer-like starch with a branched structure.The amount of amylose and amylopectin and their molecular weight are different from those of different starches. Vary between types.

  In order to improve the properties of starch for use in this invention, starches such as corn or potato starch can be obtained, for example, by acid hydrolysis with citric acid and / or polyethylene glycol (PEG) and / or excess. By reacting with a reagent such as hydrogen oxide, modification that improves the hydrophobicity can be performed. In addition, the starch can optionally be subjected to a heat treatment after treatment with PEG and / or hydrogen peroxide to reduce the amount of branching, for example at 160 to 180 ° C. for 30 to 60 minutes (hereinafter, This is called “heat-destructed starch”).

  In another preferred embodiment, the nanoparticles of this invention are encapsulated composites wherein the active compound is clarithromycin and the amphiphilic polymer is selected from the group consisting of starch, chitosan, and alginic acid such as sodium alginate. Make up body. The starch may be hydrolyzed starch, starch modified with different amounts of PEG, preferably PEG-400, and / or hydrogen peroxide, and starch that has been heat destroyed.

  In another preferred embodiment, the nanoparticles of the present invention comprise an encapsulated complex wherein the active compound is azithromycin and the amphiphilic polymer is chitosan or an alginate derivative such as propylene glycol alginic acid (Manucol ester B). Yes. In another preferred embodiment, the nanoparticles of the invention constitute an encapsulated complex in which the active substance is azithromycin and the amphiphilic polymer is polyvinyl alcohol (PVA).

  In another preferred embodiment, the nanoparticles of this invention constitute an inclusion complex where the active substance is donepezil and the amphiphilic polymer is a polysaccharide.

  Donepezil, 1-benzyl-4-((5,6-dimethoxy-1-indanone) -2-yl) methylpiperidine, and analogs thereof are described in US 4,895,841 as acetylcholinesterase inhibitors, It is useful for the treatment of various types of dementia, including Alzheimer-type senile dementia, Huntington's chorea, Pick's disease, and ataxia. Donepezil hydrochloride is a white crystalline powder that is freely soluble in chloroform, soluble in water and glacial acetic acid, faintly soluble in ethanol and acetonitrile, and hardly soluble in ethyl acetate and hexane. . Donepezil hydrochloride is available by oral administration in film-covered tablets containing 5 or 10 mg of it for hypoallergenic treatments to relieve Alzheimer-type dementia. Amorphous donepezil hydrochloride is described in patents US 5,985,864 and US 6,140,321. Recently, US 6,734,195 published that wet granulation of donepezil hydrochloride produces a stable granule that homogeneously contains amorphous donepezil hydrochloride after drying and milling.

  According to the present invention, the water-soluble nanoparticles are formed in a non-crystalline state, such as an amorphous form of donepezil hydrochloride, which is encapsulated by the amphiphilic polysaccharide and has no interaction with the enclosed amphiphilic polysaccharide. The construction of an encapsulated complex immobilized / stabilized by action is provided. In one preferred embodiment, the polysaccharide is alginic acid. In another preferred embodiment, the polysaccharide is sodium starch glycolate. In yet another embodiment, the polysaccharide is pregelatinized modified starch.

  In another preferred embodiment, the nanoparticles of the present invention comprise an active substance is an azole compound, and the amphiphilic polymer is a polysaccharide, polyacrylic acid, polyacrylic acid copolymer, polymethacrylic acid, and polymethacrylic acid. An encapsulated composite that is a copolymer of

  Azole compounds play an important role as antifungal agents in agriculture and human mycosis and as nonsteroidal antiestrogens in the treatment of postmenopausal female estrogen-responsive breast cancer. This broad use of azoles is based on the inhibition of certain pathways of the sterol synthesis system by high affinity binding to the enzymes sterol 14-demethylase and aromatase. Azole antifungal agents exhibit a wide range of antifungal activities and are used to prevent or treat fungal infections. They are therefore important tools in integrated agricultural production. According to their chemical structure, azole compounds are classified into triazoles and imidazoles; however, their antifungal activity is by the same molecular mechanism. Azole antifungal agents are widely used in agriculture and antifungal treatment of humans and veterinarians.

  According to the invention, “azole compounds” refer to imidazole and triazole compounds for human and veterinary applications or for use in agriculture.

  In one preferred embodiment, the azole compound is not limited to the triazoles terconazole, itaconazole, and fluconazole, but is the imidazole clotrimazole, miconazole, econazole, ketoconazole, thioconazole, isoconazole, oxyconazole, and Selected from azole antifungal agents used in many different antifungal formulations, including fenticazole and the like.

  In another example, azole compounds include letrozole, anastrozole, borozole, and fadrozole, which act as nonsteroidal antiestrogens and can be used to treat estrogen-responsive breast cancer in postmenopausal women. However, it is selected from azoles that are not so limited.

  In another example, the azole compound is a triazole useful for agriculture: vitertanol, cyproconazole, difenoconazole, epoxiconazole, fluquinconazole, flusilazole, flutriafol, hexaconazole, metconazole, microbuta Nyl, penconazole, propiconazole, tebuconazole, triazimephone, triazimenol, and triticonazole, and azole antifungal agents, including but not limited to imidazole, imazalyl, procoraz, and triflumizole. In yet another embodiment, the azole compound is a triazole, used as an acaricide, azocyclotine, used as a growth regulator, paclobutrazol, used as a herbicide, carfentolazole, used as a herbicide, and an insecticide. Isazophos and imidazole, non-antifungal azoles for use in agriculture, such as metazachlor, used as a herbicide.

In one more preferred embodiment, the azole compound is itraconazole, an azole drug used in the treatment of fungal infections. Itraconazole is effective against a wide range of fungi, including dermatophytes (ringworm infections), yeasts such as Candida and Malassezia infections, histoplasma, Aspergillus, coccidioidomycosis, and systemic fungal infections such as chromo-blastosmosis. It is effective. Itraconazole is available as a 100 mg capsule under the registered trademark Sporanox ^™ (Jansen-Silag). It is a white to slightly yellowish powder. It is lipophilic, insoluble in water, slightly soluble in alcohol, and freely soluble in dichloromethane. Sporanox contains 100 mg of itraconazole covered with saccharide spheres.

  In one embodiment, the amphiphilic polymer used to encapsulate the azole compound is a polysaccharide, more preferably chitosan, or starch that has been hydrolyzed or thermally destroyed, optionally together with PEG, hydrogen peroxide, or It is denatured by both. Alginic acid can also be used with certain concentrations of azole compounds (see Table 5 below).

  In another embodiment, the amphiphilic polymer used to encapsulate the azole compound is selected from the group consisting of polyacrylic acid, polyacrylic acid copolymer, polymethacrylic acid, polymethacrylic acid copolymer. . The copolymer of polyacrylic acid / polymethacrylic acid may be a copolymer of acrylic acid / methacrylic acid containing another acrylic acid / methacrylic acid derivative such as alkylacrylic acid / alkylmethacrylic acid. In one preferred embodiment, the amphiphilic polymer is polyacrylic acid. In another preferred embodiment, the amphiphilic polymer is a copolymer of acrylic acid containing butylacrylic acid in different proportions (see Table 5).

  In yet another preferred embodiment, the nanoparticles of the invention constitute an encapsulated complex in which the active substance is a taxane and the amphiphilic polymer is gelatin.

  As used herein, the term “taxane” refers to, for example, the center of a 20 carbon taxane, represented by the structural formula shown in US Pat. References are made to compounds containing frameworks. The term taxane refers to the chemotherapeutic drug taxol (generic name: paclitaxel; chemical name: 5β, 20-epoxy-1,2α, 4,7β, 10β, 13α-hexahydroxytax-111-en-9-one, 4, 10-diacetate 2-benzoate 13-ester (2R, 3S) -N-benzoyl-3-phenylisoserine) and taxotea (generic name: docetaxel) and, for example, an ester or ether at the C (7) position Substitution, hydroxy substitution at the C (10) position, and various C (2), C (9), C (14) positions, and for example, all these patents are incorporated by reference in their entirety and fully published Patents US 6,794,523, US 6,780,879, US 6,765,015, US 6,610,860, US 6 552,205, and includes a semi-synthetic derivative of the taxane having substitutions, such as the side chains as described in US 6,201,140.

Taxol is now an anti-cancer drug with the generic name “paclitaxel”, the registered trademark is “Taxol ^R ” (Bristol-Myers Squibb), originally isolated from the bark of the Pacific Yew tree Complex, multi-oxygenated diterpenes. Taxol is approved by the FDA to treat breast cancer, ovarian cancer, and lung cancer, as well as Kaposi's sarcoma associated with AIDS. Docetaxel (Taxotea-R) is a substance that is similar to paclitaxel and is obtained from the needles of yew trees and has no effect on other anticancer agents, such as advanced breast cancer and non-small cells Approved by the FDA to treat lung cancer. Paclitaxel and docetaxel are administered intravenously. Both paclitaxel and docetaxel have serious side effects. Paclitaxel is a white to greyish white crystalline powder. This natural product is highly hydrophobic and insoluble in water. One problem with the administration of taxol is its poor solubility in most pharmaceutically usable solvents; clinically used formulations include Cremophor EL (multi-ethoxylated castor oil) and excipients Containing ethanol, they have serious adverse effects. Therefore, despite the good clinical effects of paclitaxel and recognized as one of the most significant cancer drugs, the better safety and pharmacokinetics of paclitaxel in patients There is still a growing need to achieve aspects.

US 6,753,006 is stable, containing a sufficient amount of non-cremophor-free paclitaxel that can be systemically administered to humans in an amount ranging from 30 to 1000 mg / m ² , A non-aqueous formulation is published under aseptic conditions.

  According to the present invention, amorphous paclitaxel is formed in which water-soluble nanoparticles are immobilized / stabilized by non-valent interaction with non-crystalline state, for example, gelatin and enclosed gelatin. An encapsulated composite composition comprising is provided. In a preferred embodiment, vitamin B12 and / or polystyrene sulfonic acid is added to gelatin to improve the solubility of paclitaxel.

  The aqueous nanodispersions of this invention can be lyophilized and then mixed with a pharmaceutically acceptable carrier that provides a stable pharmaceutical composition.

  Pharmaceutically acceptable carriers or excipients are adapted to the type of active compound and type of formulation and are well known in the art, for example, Remington: The Science and Practice of Pharmacy (formerly Remington's Pharmaceutical Sciences). 19th ed. , 1995, can be selected from standard excipients.

  Thus, in another aspect, the present invention provides a stable pharmaceutical composition comprising a pharmaceutically acceptable carrier and the nanodispersion of this invention. This composition is intended for oral, intravenous, mucosal, and pulmonary administration. In a more preferred embodiment, the composition is for oral administration and can be in liquid or solid form. In one preferred embodiment, the tablet form exemplified herein for azithromycin is provided.

  In one preferred embodiment, the invention provides a bacterial infection comprising water-soluble nanoparticles comprising an encapsulated complex wherein the active compound is selected from the group consisting of erythromycin, clarithromycin, azithromycin and the amphiphilic polymer is a polysaccharide. It relates to a stable pharmaceutical composition for the treatment of These compositions are useful against bacterial infections treatable by the aforementioned macrolide antibiotics, particularly respiratory infections.

  In another preferred embodiment, the present invention relates to dementia or Alzheimer's disease comprising a nanodispersion of water-soluble nanoparticles comprising an encapsulated complex in which the active compound is donepezil hydrochloride and the amphiphilic polymer is a polysaccharide. It relates to a stable pharmaceutical composition for treatment.

  In a further preferred embodiment, the present invention provides that the active compound is an azole antifungal agent and the amphiphilic polymer is a polysaccharide, polyacrylic acid, polyacrylic acid copolymer, polymethacrylic acid, polymethacrylic acid copolymer. It relates to a stable pharmaceutical composition for the treatment of fungal infections, comprising a nanodispersion of water-soluble nanoparticles constituting an inclusion complex selected from the group consisting of In a more preferred embodiment, the azole antifungal agent is itraconazole and the amphiphilic polymer is polyacrylic acid, a copolymer of acrylic acid and butylacrylic acid, chitosan, and acid hydrolysis, polyethylene glycol or hydrogen peroxide and Or modified starch that has been subjected to one or more of heat treatments.

  In yet another preferred embodiment, the present invention relates to a water soluble nanostructure comprising an encapsulated complex wherein the active compound is a non-steroidal anti-estrogen azole selected from the group consisting of letrozole, anastrozole, borozole, and fadrozole. It relates to a stable pharmaceutical composition for the treatment of estrogen-responsive breast cancer, consisting of nano-dispersion of particles.

  In a further preferred embodiment, the invention relates to the treatment of cancer comprising a nanodispersion of water-soluble nanoparticles constituting an encapsulation complex in which the active compound is a taxane, most preferably paclitaxel and the amphiphilic polymer is gelatin. Related to a stable pharmaceutical composition.

  The invention will now be illustrated by the following non-limiting examples.

Example 1 General Method for Production of Nanoparticles Comprising Encapsulated Composites For the preparation of nanoparticles of this invention, the following general method is performed:
(I) preparation of a molecular solution of an amphiphilic polymer in water;
(Ii) preparation of a molecular solution of the active compound in an organic solvent;
(Iii) A cooled solution of the active compound (ii) is added dropwise to the polymer solution (i) heated at a temperature 5-10 ° C. above the boiling point of the organic solvent of (ii) under constant mixing;
(Iv) Evaporation of the organic solvent, resulting in the desired nanodispersion of the nanoparticles that make up the encapsulated complex of active compound encapsulated in the amphiphilic polymer.

  This method is performed as previously described by the inventor in US 2003/0129239 referenced above, using a chemical reactor written therein, or with appropriate modifications of the aforementioned apparatus. It may be. According to the present invention, during the process of forming soluble nano-sized particles, the polymer is added to an aqueous solution, preferably water, so as to form a polymer solution within the first vessel of the chemical reactor. In addition, ingredients are added to adjust the pH and ionic force levels of this solution as needed, based on parameters determined through the algorithm used to select the active compound and polymer. Active compounds that are water-insoluble (lipophilic) or water-soluble (hydrophilic) compounds are contained in a second vessel of the chemical reactor. A solution of the active lipophilic or hydrophilic compound in a non-aqueous solvent (or mixture of solvents) is referred to as a “carrier”. The rate at which the carrier is poured or added to the polymer solution is controlled by one or more adjustable faucets to ensure that the organic solution added to the polymer solution is at a concentration below 3%. .

  The active compound solution is formed when the polymer solution is heated and the vapor from the heated polymer solution liquefies and dissolves the active compound present in the second container. The active compound solution (in the carrier) is then mixed with the polymer solution so as to form a dispersed phase in the emulsion or suspension. Within the chemical reactor, the emulsion is directed to a region of turbulence provided by a stirrer (more precisely, a nanostirrer) that results in the formation of nano-sized active compound molecules in the emulsion or suspension. And added. This region of turbulence is referred to as the “region of action” or “region of interaction”. The emulsion or suspension added to the turbulent region has a Reynolds number Re> 10,000. The emulsion thus becomes a “nanoemulsion” or “nanosuspension” with particles ranging from approximately 1 to approximately 1000 nm. Particle production can also be extended to include small micron sized particles, and these particles are suitable for several uses and are covered by the present invention.

  In a nanoemulsion or nanosuspension there is a dispersed solvent comprising a polymer solution and a dispersed phase that constitutes a solution of the active compound in a carrier. However, this two-phase nanoemulsion or nanosuspension is unstable. Evaporation of the support causes dispersed phase particles in the size range of approximately 1 to approximately 1000 nm. The polymer molecules in the polymer solution then enclose or package, and more appropriately enclose, the active compound that remained in the dispersed phase particles after evaporation of the carrier, and thus was encapsulated by the hydrophilic polymer in the encapsulation complex. Form a homogeneous nano-sized dispersion of water-insoluble lipophilic compounds. The remaining carrier is then removed by vacuum drying or other suitable drying technique (eg lyophilization, vacuum distillation). The result of the algorithm used to select the appropriate active compound and polymer for the formation of the emulsion or suspension, and the resulting complex, generally remains after evaporation of the carrier. There is no free polymer. Following evaporation of the carrier, a stable encapsulated complex is composed of amorphous and / or partial crystals, or crystalline actives.

Example 2 Preparation of Modified Starch For use of this invention, it is desirable to use starch that is mostly linear, i.e. α-D-1, such that the anhydroglucose units form a linear chain. It is a starch containing a high content of amylose, which is a constituent of starch linked by 4 glycosidic bonds, and a low content of amylopectin, a constituent of starch having a polymer-like branched structure. The levels of amylose and amylopectin and their molecular weight vary between different starch types.

  In order to improve its properties for use in the present invention, corn or potato starch is rendered hydrophobic by, for example, acid hydrolysis and / or reaction with reagents such as polyethylene glycol and / or hydrogen peroxide. It can be modified by improving. In addition, the starch can be subjected to a heat treatment to reduce the amount of branching, for example at 160 to 180 ° C. for about 30 to 60 minutes (hereinafter referred to as “heat destroyed starch”).

  For modification, various amounts of potato starch and distilled water are placed in the reaction vessel (C p.st = potato starch concentration, X1 in Table 1) and given the desired pH (range 2 to 5). Citric acid was added under mixing until (X2, Table 1). The resulting suspension was heated from room temperature to 70 to 95 ° C. for approximately 10 to 20 minutes with continuous stirring until a homogeneous opaque body (hydrolyzed starch) was obtained. The resulting object was exposed to 160-180 ° C. in an autoclave for time X3 (minutes). Under these conditions, the starch network is partially or fully converted into a linear, slightly branched polymer that dissolves in water. The object was cooled to below 100 ° C. (heat destroyed starch).

  For some samples, PEG-400 was added (amount X4,% to starch, Table 1) and the resulting mixture was at 160-180 ° C. in an autoclave for time X5 (Table 1). Heated and then cooled to below 100 ° C. (PEG-modified pyrolyzed starch). The turbidity (FTU, in formazine turbidity units) and viscosity (molecular weight, MW) of the solution were measured. The results are shown in Table 1. A solution suitable for further use should be clear or opalescent and preferably have a turbidity in the range of 20 to 40 FTU. Furthermore, the molecular weight (MW) of the modified starch is calculated by intrinsic viscosity measurements. Acceptable MW (reflected by intrinsic viscosity) values are up to approximately 100,000 and depend on the active compound to be complexed.

Example 3 Preparation of Nanoparticles Constructing Encapsulated Complex of Clarithromycin Encased in Modified Starch This example is not part of the present invention and is intended to illustrate the process of nanoparticle preparation .

For the preparation of amphiphilic polymers, potato starch with a molecular weight of (5-10) × 10 ⁴ is dissolved in distilled water, first heated at 160 to 180 ° C., and a starch: PEG-400 ratio of 2 Using a range between 1: 1 and 4: 1, the pH of the solution is adjusted to 6.5 or lower with citric acid, the temperature is 160 to 180 ° C., the denaturation time is 60 to 180 minutes, Modified with PEG-400 as described in 2. A solution of clarithromycin in methyl acetate or dichloromethane was prepared.

  An aqueous solution of modified starch was placed in a reaction vessel and heated to 60 ° C. while mixing with a homogenizer at a speed of 10,000 revolutions / minute or more. After the starch solution temperature reached 60 ° C., the solution of clarithromycin was added thereto at a rate of about 1 ml / sec. The homogenizer speed was also at least 10,000 revolutions / minute. Clarithromycin interacted with the modified starch to make nanoparticles and the organic solvent was evaporated and concentrated directly in a concentrator. After all clarithromycin has interacted with the polymer and dissolved as a clarithromycin-starch encapsulated complex, the remaining organic solvent is vacuum dried with continued mixing to form the clarithromycin-starch encapsulated complex. The aqueous solution of nanoparticles was cooled to 30-35 ° C.

  The turbidity and viscosity of the cooled aqueous solution of nanoparticles were measured at a defined storage time to assess dispersion stability. The turbidity values for the nanodispersions of some clarithromycin-starch inclusion complexes are shown in Table 2. A stable nanodispersion shows a turbidity value that remains unchanged throughout the time. The presence of the crystalline phase and the size of the composite particles were determined.

EXAMPLE 4 Physical Characteristics of Additional Clarithromycin-Polymer Encapsulation Complex An additional clarithromycin hydrophilic encapsulation complex is prepared by dissolving clarithromycin in methyl acetate or dichloromethane and hydrolyzing the polymer into hydrolyzed potato starch, alginic acid. , Chitosan, or polyvinyl alcohol (PVA) was prepared according to the method described in Example 1.

  Table 3 below shows the properties of various such complexes. Shown in Table 3 are the complex name (Exp., First column), polymer name and concentration (%), drug concentration, pH, and ALV size and size distribution (nm), Physicochemical analysis of various composite nanoparticles, including HPLC (concentration and solubility), and in some cases, powder X-ray analysis for crystal phase determination, and the like. Complex size measurements were performed using ALV technology and powder X-ray analysis was performed as described above in Example 6.

  FIG. 1 illustrates the size distribution of the nanoparticles that make up the clarithromycin hydrophilic inclusion complex (# 10-134 in Table 3) in 1% chitosan, which was approximately 838 nm in size.

  As shown in Table 3, nanoparticles (size less than 1000 nm) can be prepared using polymers such as hydrolyzed starch, alginic acid, and chitosan from different sources, but when using PVA, the particles are 1600 nm. This clearly indicates that PVA is not useful for the preparation of nanoparticles containing macrolides because the particles are crystalline and not amorphous.

  The results in Table 3 show that when the macrolide antibiotic clarithromycin, a poorly soluble hydrophobic compound, is surrounded by an amphiphilic polymer, the resulting inclusion complex is hydrophilic. Yes. Using the matching technique described in this application, clarithromycin was made hydrophilic even when surrounded by a variety of polymers that met the matching parameters, such as alginic acid, PVA, and chitosan. The results show that 2% alginate conjugated with 10 mg / ml clarithromycin at pH 5.5 results in 530 nm ALV size distribution analysis nanoparticles; 2% PVA conjugated with clarithromycin at pH 6 is 1600 nm ALV. 1% chitosan (Fluka) bound to clarithromycin at pH 4 to 6 results in 165 nm ALV; 1% chitosan (Fluka) bound to clarithromycin at pH 4 becomes 321 nm ALV; 1% chitosan (Fluka) bound to clarithromycin at pH 6 resulted in 660 nm ALV; and 1% chitosan (Sigma) bound to clarithromycin at pH 5 showed 838 nm ALV. These results show that, using the teachings of the specification (lipophilic / amphiphilic polymer matching technology), poorly soluble hydrophobic antibiotics (clarithromycin) can be used with various amphiphilic polymers (ie, alginic acid, PVA, and then chitosan), indicating that the resulting inclusion complex can become hydrophilic in water.

  It was also found that unchanged spherical nanoparticles of clarithromycin-hydrolyzed starch inclusion complex IC-76 were obtained immediately and even 5 weeks after preparation (data not shown) ).

Example 5 Controlled Release of Clarithromycin from Clarithromycin-Polymer Encapsulated Complex via Dialysis This experiment was performed with a 3500D molecular weight cut-off as described in US 2003/0129239 (in its Example 7). Value of cellulose dialysis membrane (SnakeSkin ^™ dialysis tubing, Pierce Chemical Co., product no.

Three dosage forms of clarithromycin were tested:
1. Commercial clarithromycin (10 mg / ml), pH 4 dissolved in water
2. Clarithromycin complex in 2% PVA of Table 3, pH = 6.0 (S-Clari # 34), with an initial calculated concentration of 10 mg / ml
3. Clarithromycin complex in 1% chitosan (IC-135 in Table 3), pH = 6.5 (S-Clari # 135) with an initial calculated concentration of 10 mg / ml

  Each dosage form (2 ml) was placed in a dialysis bag in a glass bottle with 100 ml water, pH = 4 (titrated with 20% citric acid). Dialysis was performed for up to 6 hours with constant stirring at 23 ± 2 ° C. Samples of external buffer (1 ml) were taken every 5 hours after the start of dialysis to analyze drug release. The volume of the external liquid was constant 100 ml. The concentration of clarithromycin in the outer (outside of the bag) and inner (in the bag) liquid and the sample taken was determined by HPLC. The results shown in FIG. 2 show that the release of clarithromycin from the PVA complex (S-Clari # 34, square) is faster than that of the commercial dosage form (Clari, diamond), while the chitosan complex The release of clarithromycin from the body (S-Clari # 135, triangle) has been shown to be significantly slower than that of the commercial dosage form. This indicates that the nanodispersion with chitosan is capable of withstanding the release of clarithromycin and is more suitable for the preparation of inclusion complexes with macrolides. As shown in Table 3, the complex with PVA was 1600 nm in size, not in the nano-range, and therefore could not withstand release.

Example 6 Physical Measurements and Properties of Various Azithromycin Hydrophilic Encapsulation Complexes An encapsulated complex of a specific macrolide antibiotic, azithromycin, in which azithromycin is dissolved in methyl acetate or dichloromethane and the polymer is alginic acid, manucol ester B (alginic acid Derivatives), chitosan or PVA and were prepared by the method described in Example 1.

  Table 4 below shows the properties of such various composites. Shown in Table 4 are the complex name (Exp., First column), polymer name and concentration (%), drug concentration, pH, and ALV size and size distribution (nm), Physicochemical analysis of various composite nanoparticles, including HPLC (concentration and solubility). Measurement of the size of the encapsulated complex was performed using an ALV-particle sizer (ALV-Laser GmbH, Langen, Germany) with a resolution of 3 to 3000 nm. ALV is a direct light scattering technique used to measure average particle size. The experiment was performed using a laser powered non-invasive backscatter = high performance particle sizer (ALV-NIBS / HPPS).

  FIG. 3 shows the size distribution of the nanoparticles that make up the azithromycin hydrophilic inclusion complex (# 10-148 / 2 in Table 4) in 1% chitosan with a size of approximately 362 nm. Furthermore, the azithromycin in these particles was found to be amorphous, as shown in the lower line of FIG. 4, and the amorphous was found to be stable for at least 10 months. It was. FIG. 4 shows the X-ray spectrum of a sample of azithromycin-chitosan inclusion complex (lower line) after 10 months, compared to commercially available azithromycin (upper line).

  The results in Table 4 indicate that when the macrolide antibiotic azithromycin, a poorly soluble hydrophobic compound, is surrounded by an amphiphilic polymer, the resulting inclusion complex is hydrophilic. Using the matching techniques described herein, azithromycin becomes hydrophilic when surrounded by a variety of polymers that meet the matching parameters, such as alginic acid, PVA, manucol ester B, and chitosan. The result is that 2% PVA conjugated with 10 mg / ml azithromycin results in a nanoparticle size distribution analysis of 5 nm ALV; 4% manucol ester B conjugated with azithromycin results in 330 nm ALV; binds azithromycin 1% PVA resulted in 5 nm ALV and 350 nm in another formulation process; 2% alginic acid coupled with azithromycin resulted in 1600 nm ALV and 1060 nm in another formulation process; 1% chitosan coupled with azithromycin (Sigma) indicates 510 nm ALV, and 752 and 362 nm for other formulation processes. These results are consistent with those described above using clarithromycin, and using the teachings herein (lipophilic / amphiphilic polymer matching technology), poorly soluble hydrophobic antibiotics (azithromycins) ) Are surrounded by various amphiphilic polymers (ie, alginic acid, PVA, manucol ester B, and chitosan), indicating that the resulting inclusion complex becomes hydrophilic in water. .

Example 7 Physical Measurements and Characteristics of Various Itraconazole Hydrophilic Encapsulation Complexes An azole antifungal itraconazole encapsulation complex was prepared by dissolving itraconazole in methyl acetate or dichloromethane and hydrolyzing the polymer, potato starch Prepared according to the method described in Example 1, which is potato starch, alginic acid, chitosan, polyacrylic acid, and a copolymer of acrylic acid and butylacrylic acid.

  Table 5 below shows the properties of various such itraconazole hydrophilic inclusion complexes. FIG. 5 shows the size distribution of the nanoparticles that make up the itraconazole hydrophilic inclusion complex (# 23-120) in heat-destructed potato starch with a size of approximately 414 nm.

The results in Table 5 indicate that when the antifungal drug itraconazole, an insoluble compound, is surrounded by an amphiphilic polymer, the resulting inclusion complex is hydrophilic. Using the matching techniques described in this application, itraconazole becomes hydrophilic when surrounded by various polymers that meet the matching parameters such as pyrolyzed starch, alginic acid, chitosan combined with H ₂ O ₂ or PEG modification. Become sex. The result was that 5% pyrolyzed starch + 1.25% H ₂ O ₂ + 1.25% PEG bound to 5 mg / ml itraconazole resulted in 382 nm ALV; bound to 5 mg / ml itraconazole 5% pyrolyzed starch + 0.625% H ₂ O ₂ + 1.25% PEG results in ALV of 640 nm; 5% pyrolyzed starch + 1% H ₂ O ₂ + 2% PEG is 414 nm 5% pyrolyzed starch + 1% H ₂ O ₂ + 1% PEG conjugated with 5 mg / ml itraconazole resulted in 793 nm ALV; 2% conjugated with 20 mg / ml itraconazole Alginic acid results in an ALV of 180 nm with 10 mg / ml itraconazole When bound, it results in an ALV of 180 nm; 1% chitosan (Fluka) bound to ˜8 mg / ml itraconazole indicates a result of 120 nm ALV. These results show that, using the teachings of this specification (lipophilic / amphiphilic polymer compatibility technology), it is possible that the insoluble antifungal drug itraconazole can be used in various amphiphilic polymers (ie, H ₂ O ₂ It is shown that the encapsulated complex resulting becomes hydrophilic in water when surrounded by pyrolyzed starch, alginic acid, and chitosan combined with modification.

  Differential scanning calorimetry (DSC) was performed using a TA Instruments 2010 module and 2100 system controller to probe the crystallinity of the complex. The test was performed from −50 to 200 ° C. at a scan rate of 10 degrees / minute. FIGS. 6A-B provide diagrams of the respective results of itraconazole crystals and itraconazole complexes prepared in Experiment IT-56 (see Table 5). Itraconazole crystals are melted at a specific melting point, whereas itraconazole complexes are not melted at a specific melting point.

Example 8 Physical Measurement and Characterization of Various Paclitaxel Hydrophilic Encapsulation Complexes The inclusion complex of the anticancer drug paclitaxel has different molecular weights with paclitaxel dissolved in methyl acetate or dichloromethane and the polymer with or without vitamin B12 added. Prepared by the method described in Example 1. Polyvinylpyrrolidone (PVP or povidone, such as Kollidon ^™ ) or polystyrene sulfonic acid can also be added to improve the solubility of paclitaxel. Polystyrene sulfonic acid can also be used alone to solubilize paclitaxel.

  Table 6 below shows the properties of various such paclitaxel hydrophilic inclusion complexes. FIG. 7 shows the size distribution of the nanoparticles that make up the paclitaxel hydrophilic inclusion complex in gelatin (70-100 kD, 1 mg / ml vitamin B12) (# 25-85) with a size of approximately 179 nm. .

  The results in Table 6 show that when the anticancer drug paclitaxel, an insoluble compound, is surrounded by an amphiphilic polymer, the resulting encapsulated complex is hydrophilic. Using the adaptation techniques taught by this application, paclitaxel is described in this application at various concentrations (eg, 0.872 mg / ml, 0.646 mg / ml, 3.781 mg / ml and 0.925 mg / ml). It becomes hydrophilic when surrounded by gelatin (optionally supplemented with vitamin B12 excipients) that is compatible with the matching parameters.

Example 9 Physical measurements and properties of various donepezil hydrophilic inclusion complexes Donepezil hydrochloride inclusion complex was prepared by dissolving donepezil hydrochloride in methyl acetate or dichloromethane and the polymers were modified corn starch, alginic acid, and starch glycolic acid Prepared by the method described in Example 1 which is sodium.

  Table 7 below shows the properties of various such donepezil hydrochloride hydrophilic inclusion complexes. FIG. 8 shows the size distribution of the nanoparticles that make up the donepezil hydrochloride hydrophilic inclusion complex (# LG-7-51) in modified corn starch with a size of approximately 600 nm.

Example 10 Oral Absorption of Nano-sized Water-Soluble Particles of Azithromycin and Azithromycin Composition Oral absorption of water-soluble nano-sized particles comprising an encapsulated complex of 1% azithromycin and 1% chitosan to allow absorption In order to evaluate the contribution of physical form, it was investigated in a preclinical model associated with rats compared to a composition containing commercially available azithromycin (azenyl).

  Azithromycin (50 mg / kg) was administered to male Sprague-Dawley rats (a set of 5 individuals) by feeding tube, 250-280 g. Blood samples were collected at regular times after administration (between 1 and 24 hours) and serum was prepared for analysis. At the end of the experiment, all rats were sacrificed by an overdose (100 mg / kg) in a set of five individuals.

The concentration of drug in rat serum (0.1 ml) was determined by LC-MS. Samples and calibration curves were prepared as follows: 50 μl of sample was mixed with 50 μl of control serum to obtain a total volume of 100 μl of serum. The diluted sample was extracted with methyl t-butyl ether, reconstituted in 40% aqueous acetonitrile after removing the solvent. Analysis was performed by LC-MS using an Agilent 1100 HPLC system with positive pressure mode atmospheric pressure electrospray ionization. Azithromycin concentration was quantified by comparison with a calibration curve ranging from 20 to 2000 ng / ml, prepared using blank rat serum with azithromycin. A concentration plot (not shown) is used to determine the timing of the maximum concentration (C _max ) and to assess the overall absorption of the drug (as reflected by the area under the concentration curve (AUC)). It was broken.

A summary of the major pharmacokinetic findings is shown in Table 8. These findings indicate that nano-sized water-soluble particles, which are the same amount as azithromycin as azenyl, improve the maximum concentration obtained (C _max ) and the overall amount of azithromycin absorption (reflected by AUC). Show. In addition, concentrations in the lung are particularly improved, whereas concentrations in other organs are improved to a lower extent. Furthermore, there was no change in the time to reach the maximum concentration.

The stability of azithromycin particles, followed by compression, and compatibility with tablet excipients was evaluated by comparing the absorption of azithromycin with the absorption of the complex prior to tablet preparation. Tablets were prepared by lyophilization of the complex followed by mixing with excipients that meet standard conditions. Tablets were formed by application of 1 ton / cm ² pressure. Prior to administration to rats, the tablets were dissolved in water. Then, azithromycin (50 mg / kg) was administered to male Sprague-Dawley rats (a set of 5 individuals) by feeding tube, 250-280 g. Pharmacokinetic studies for oral administration were performed according to the method described above.

  The concentration of drug in rat serum was analyzed according to the method described above. A plot of serum concentration is shown in FIG. In this figure, azenil is a commercial formulation that is on the market for azithromycin, while lots 28-39 and 28-59 are nano-components that make up a complex of 1% chitosan and 1% azithromycin. Size 28 is a solution of water soluble particles and Tab 28-59 is a tablet prepared from lot 28-59 dissolved in water just prior to administration. It is clear from this figure that the maximum concentration of azithromycin generally reached the same time in all of the preparation methods. However, the absorption of azithromycin from the particles is always greater than that of commercial formulations. Thus, as indicated above, improved absorption is clearly associated with formulations consisting of water-soluble nanosized particles. Furthermore, the calculated area under the curve for tablets is only about 10% smaller than the area of a solution consisting of water-soluble nanosized particles. Therefore, the steps taken to prepare the tablet do not adversely affect the nano-sized particles.

Example 11 Oral Absorption of Itraconazole Nano-sized Water-soluble Particles Itraconazole nano-sized water-soluble particles comprising itraconazole encapsulated complex of acrylic acid and butylacrylic acid copolymer (# IT-50, Table 5) Oral absorption consists of itraconazole mixed with polyacrylic acid that does not form nanoparticles and vortexed in detail in a preclinical model associated with rats to assess the contribution of physical form to enable absorption It was compared with oral absorption of itraconazole in the composition.

  Itraconazole (50 mg / kg) was administered to male Sprague-Dawley rats (a set of 5 individuals) by feeding tube, 250-280 g. At certain times after administration (between 1 and 24 hours), blood samples were collected and serum was prepared for analysis. At the end of the experiment, all rats were sacrificed by an overdose (100 mg / kg) in a set of five individuals.

Drug concentrations in rat serum (0.1 ml) were basically determined by HPLC using the method described by Yoo et al. (2002) Arch Pharm Res 25: 387-391. Samples and calibration curves were prepared as follows: Samples were mixed with an equal volume of acetonitrile to obtain a total volume of 400 μl. KCl was added to the sample to precipitate the protein, and the resulting itraconazole in the supernatant was applied to a Merck HPLC system. Itraconazole concentration was quantified by comparing to a calibration curve ranging from 20 to 1000 ng / mL, prepared using blank rat serum mixed with itraconazole. A concentration plot (not shown) is used to determine the timing of the maximum concentration (C _max ) and to assess the overall absorption of the drug (as reflected by the area under the concentration curve (AUC)). It was broken.

A summary of the major pharmacokinetic findings is shown in Table 9. These findings indicate that administration of nano-sized water-soluble particles in the same amount as itraconazole doubles the improved maximum blood concentration (C _max ) of both itraconazole and its active hydroxylated metabolite (hydroxyitraconazole) And it shows increasing the total amount of itraconazole absorbed, reflected by the area under the curve (AUC) of both itraconazole and its active hydroxylated metabolite.

Size distribution of nanoparticles comprising clarithromycin-chitosan inclusion complex (# 10-134, Table 3) that was approximately 838 nm in size as measured by light diffraction (ALV) Particles constituting clathromycin and PVA inclusion complex (S-Clari # -34, Table 3), or nanoparticles constituting clarithromycin and chitosan inclusion complex (S-Clari # 135, Table 3) In vitro release of commercial clarithromycin (Clari) through a dialysis membrane Size distribution of nanoparticles comprising the azithromycin-chitosan inclusion complex (# 10-148 / 2, Table 4), which was approximately 362 nm in size as measured by light diffraction (ALV) X-ray spectrum of a sample of azithromycin-chitosan inclusion complex after 10 months (bottom line) compared to commercial azithromycin (top line) Size distribution of the nanoparticles that make up the itaconazole-modified starch inclusion complex (# 23-120, Table 5), which was approximately 414 nm in size as measured by light diffraction (ALV) Differential scanning calorimetry (DSC) analysis of commercially available crystalline itaconazole (12A) Differential scanning calorimetry (DSC) analysis of itaconazole-polyacrylic acid inclusion complex (# IT-56, Table 5) Size distribution of nanoparticles comprising paclitaxel-gelatin inclusion complex (# 25-85, Table 6), which was approximately 179 nm in size as measured by light diffraction (ALV) Size distribution of nanoparticles comprising donepezil-modified starch inclusion complex (# LG-7-51, Table 7), which was approximately 600 nm in size as measured by light diffraction (ALV) Oral absorption of the following material in a preclinical model for rats: the commercial product of azithromycin (Azenil), a flowable formulation of nanoparticles of azithromycin-chitosan inclusion complex (lots 28 to 39 and 28) And from lots 28 to 59, further formulated into tablet form.

Claims (32)

A nano-dispersion of water-soluble and stable nano-sized particles comprising a hydrophilic inclusion complex, consisting essentially of an active compound enclosed in an amphiphilic polymer, comprising: The active compound is in an amorphous state, and the encapsulation complex is stabilized by an unvalent interaction between the active compound and the surrounding amphiphilic polymer, and the encapsulation complex is: A nanodispersion selected from the group consisting of:
(I) an encapsulation complex in which the active compound is clarithromycin and the amphiphilic polymer is alginic acid or chitosan, or the active compound is azithromycin and the amphiphilic polymer is a polysaccharide or polyvinyl alcohol;
(Ii) an encapsulated complex wherein the active compound is donepezil hydrochloride and the amphiphilic polymer is a polysaccharide;
(Iii) the active compound is an azole compound, and the amphiphilic polymer is selected from the group consisting of polysaccharides, polyacrylic acid, copolymers of polyacrylic acid, polymethacrylic acid, and copolymers of polymethacrylic acid And (iv) an encapsulation complex in which the active compound is a taxane and the amphiphilic polymer is gelatin.   The nanodispersion according to claim 1, wherein the nanoparticles constitute an encapsulated complex in which the active compound is azithromycin and the amphiphilic polymer is a polysaccharide.   3. The nano-dispersion of claim 2, wherein said polysaccharide is starch, or starch that has been modified to improve its hydrophilicity or to reduce its branching, or both.   4. The nano-dispersion of claim 3, wherein said starch has been modified by one or more of the following treatments: acid hydrolysis, reaction with polyethylene glycol or hydrogen peroxide, or heat treatment.   The nanodispersion of claim 2, wherein the polysaccharide is chitosan or propylene glycol alginic acid.   The nanodispersion of claim 1, wherein the nanoparticles comprise an encapsulated complex in which the active compound is azithromycin and the amphiphilic polymer is polyvinyl alcohol (PVA).   The nanodispersion of claim 1, wherein the nanoparticles constitute an encapsulated complex wherein the active compound is donepezil hydrochloride and the amphiphilic polymer is a polysaccharide.   8. The nanodispersion of claim 7, wherein the polysaccharide is selected from the group consisting of alginic acid, sodium starch glycolate, and pregelatinized modified starch.   The nanoparticles constitute an encapsulated complex in which the active compound is an azole compound and the amphiphilic polymer is a polysaccharide, polyacrylic acid, polyacrylic acid copolymer, polymethacrylic acid, polymethacrylic acid copolymer, The nanodispersion of claim 1.   10. The nanodispersion of claim 9, wherein the azole compound is an imidazole or triazole compound for human or veterinary application or agricultural use.   11. The azole antifungal agent of claim 10, wherein the azole compound is an azole antifungal agent selected from the group consisting of telconazole, itraconazole, fluconazole, clotrimazole, miconazole, econazole, ketoconazole, thioconazole, isoconazole, oxyconazole, and fenticazole. Nano dispersion.   11. The nanodispersion of claim 10, wherein the azole compound is a non-steroidal antiestrogen selected from the group consisting of letrozole, anastrozole, borozole, and fadrozole.   The azole compound is vitertanol, cyproconazole, difenoconazole, epoxiconazole, fluquinconazole, flusilazole, flutriafol, hexaconazole, metconazole, microbutanyl, penconazole, propiconazole, tebuconazole, triazimephone, 11. The nanodispersion of claim 10, which is an agriculturally useful azole antifungal agent selected from the group consisting of triadimenol and triticonazole, imazalil, procollaz, and triflumizole.   11. The nanodispersion of claim 10, wherein the azole compound is a non-antifungal azole for use in agriculture selected from the group consisting of azocyclotin, paclobutrazol, carfentolazole, isazophos, and metazachlor.   The amphiphilic polysaccharide is selected from the group consisting of chitosan and starch modified by one or more subsequent treatments, i.e. acid hydrolysis, reaction with polyethylene glycol or hydrogen peroxide, or heat treatment, Nanodispersion according to any of claims 9 to 14.   The nanodispersion according to any one of claims 9 to 14, wherein the amphiphilic polymer is polyacrylic acid or a copolymer of acrylic acid and butylacrylic acid.   The azole compound is itraconazole and the amphiphilic polymer is polyacrylic acid, a copolymer of acrylic acid and butylacrylic acid, chitosan, and one or more of the following treatments: acid hydrolysis, polyethylene glycol or 10. Nanodispersion according to claim 9, selected from the group consisting of starch modified by reaction with hydrogen oxide or heat treatment.   The nanodispersion of claim 1, wherein the nanoparticles comprise an encapsulated complex wherein the active compound is a taxane and the amphiphilic polymer is gelatin.   19. The nanodispersion of claim 18, wherein the taxane is paclitaxel, docetaxel, or a semisynthetic derivative of a taxane.   20. The nanodispersion of claim 19, wherein a reagent selected from the group consisting of vitamin B12, polyvinyl pyrrolidone, and poly (4-styrene sulfonic acid) is added to gelatin. (I) preparing a molecular solution of an amphiphilic polymer in water;
(Ii) preparation of a molecular solution of said active compound in an organic solvent, wherein the active compound is selected from the group consisting of azithromycin, donepezil hydrochloride, azole compound, and taxane;
(Iii) A cooled solution of the active compound (ii) is added dropwise to the polymer solution (i) heated at a temperature 5-10 ° C. above the boiling point of the organic solvent of (ii) under constant mixing;
(Iv) Evaporation of the organic solvent, resulting in a nanodispersion comprising nanoparticles composed of encapsulated complexes in which the active compound is encapsulated in the amphiphilic polymer via non-valent interactions ,
A process for the preparation of a nanodispersion according to claim 1 comprising the steps of:   A stable pharmaceutical composition comprising the nanodispersion of claim 1 and a pharmaceutically acceptable carrier.   24. The stable pharmaceutical composition of claim 22 for oral administration.   24. The stable pharmaceutical composition of claim 22 or 23, which is in liquid or solid form.   25. The stable pharmaceutical composition of claim 24, which is in the form of a tablet.   23. A stable pharmaceutical composition according to claim 22 for the treatment of bacterial infections consisting of a nanodispersion of water-soluble nanoparticles constituting an encapsulation complex in which the active compound is azithromycin and the amphiphilic polymer is a polysaccharide or polyvinyl alcohol. object.   23. The stable treatment of claim 22 for the treatment of dementia and Alzheimer's disease, comprising a nanodispersion of water soluble nanoparticles comprising an encapsulated complex in which the active compound is donepezil hydrochloride and the amphiphilic polymer is a polysaccharide. Pharmaceutical composition.   An encapsulant wherein the active compound is an azole antifungal agent and the amphiphilic polymer is selected from the group consisting of polysaccharides, polyacrylic acid, copolymers of polyacrylic acid, polymethacrylic acid, and copolymers of polymethacrylic acid 23. The stable pharmaceutical composition of claim 22 for the treatment of fungal infections, comprising a nanodispersion of water soluble nanoparticles that make up the complex.   The azole antifungal agent is itraconazole and the amphiphilic polymer is polyacrylic acid, a copolymer of acrylic acid and butylacrylic acid, chitosan, and one or more of the following treatments: acid hydrolysis, polyethylene glycol or 30. The stable pharmaceutical composition of claim 28, selected from the group consisting of reaction with hydrogen peroxide or starch modified by heat treatment.   Of an estrogen-responsive breast cancer comprising a nano-dispersion of water-soluble nanoparticles comprising an encapsulated complex, wherein the active compound is a non-steroidal anti-estrogen azole selected from the group consisting of letrozole, anastrozole, borozole and fadrozole 24. The stable pharmaceutical composition of claim 22 for treatment.   23. A stable pharmaceutical composition according to claim 22 for the treatment of cancer, consisting of a nanodispersion of water-soluble nanoparticles constituting an inclusion complex wherein the active compound is a taxane and the amphiphilic polymer is gelatin.   32. The stable pharmaceutical composition of claim 31 for the treatment of cancer, wherein the taxane is paclitaxel.

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