JP2011178729A - Nanoparticle, nanocomposite particle and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a nanoparticle, a nanocomposite particle and a method for producing the same, provided that the nanoparticle is excellent in redispersibility in water. <P>SOLUTION: The nanoparticle comprises a material to be delivered, a biodegradable polymer and an amino acid and has a volume-average particle size of 1-900 nm. The biodegradable polymer is chosen from polylactic acid, polyglycolic acid, poly(lactic acid-glycolic acid) and polycyanoacrylate. The amino acid is preferably a basic amino acid. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to nanoparticles, and nanocomposite particles and methods for producing the same.

So-called nanoparticles with an average particle size of less than 1 μm have characteristics such as an extremely large specific surface area compared to conventional micron-sized fine powders, and are thus very useful for pharmaceutical technology applications in the pharmaceutical field. Expectation is put.
Nanoparticles containing a delivery target are expected to have various advantages in a drug delivery system (DDS). For example, in the case of injection administration, side effects are expected to be reduced because the drug is selectively collected in the liver, lungs, or inflamed sites and released. In addition, when administered orally, it is expected that absorption of a hardly absorbable physiologically active substance will increase. Furthermore, in the case of transdermal administration, the physiologically active substance can be administered locally.

  Several techniques have been disclosed for nanoparticles containing deliverables. For example, a composition for intravenous injection containing nanoparticles in which a prostanoid or the like is encapsulated in a polylactic acid-glycolic acid copolymer (hereinafter sometimes referred to as “PLGA”) and lecithin is attached to the surface is disclosed. (For example, refer to Patent Document 1). Moreover, the nanoparticle which included the anticancer drug in PLGA is disclosed (for example, refer patent document 2).

JP 2003-342196 A Special table 2009-512682 gazette

However, the nanoparticles described in Patent Documents 1 and 2 may not be sufficiently redispersible in water.
An object of the present invention is to provide nanoparticles excellent in redispersibility in water, nanocomposite particles containing the nanoparticles, and a method for producing the same.

A first aspect of the present invention is a nanoparticle containing a delivery target, a biodegradable polymer, and an amino acid, and having a volume average particle diameter of 1 nm to 900 nm.
The amino acid is preferably a basic amino acid. The biodegradable polymer is preferably a biodegradable polymer containing at least one selected from polylactic acid, polyglycolic acid, poly (lactic acid-glycolic acid) and polycyanoacrylate, and the volume average particle diameter is Is more preferably 10 to 700 nm.

  The second aspect of the present invention is nanocomposite particles containing at least one of the above-mentioned nanoparticles and having a volume average particle diameter of 1 μm or more and 500 μm or less. The nanocomposite particles preferably further contain at least one saccharide.

  A third aspect of the present invention is a dispersion in which a solution containing a delivery target, a biodegradable polymer, and an organic solvent is mixed with water to obtain a dispersion of nanoparticles containing the delivery target and the biodegradable polymer. It is a manufacturing method of the nanocomposite particle | grains including the process and the drying process which removes water from the dispersion of the said nanoparticle in presence of an amino acid.

  ADVANTAGE OF THE INVENTION According to this invention, the nanoparticle excellent in the redispersibility to water, the nanocomposite particle containing this nanoparticle, and its manufacturing method can be provided.

<Nanoparticles>
The nanoparticle of the present invention contains at least one kind to be delivered, at least one kind of biodegradable polymer, and at least one kind of amino acid, and has a volume average particle diameter of 1 nm to 900 nm.
In the nanoparticle of the present invention, a nanoparticle having excellent redispersibility in water can be formed by combining a nanoparticle containing a delivery target and a biodegradable polymer and an amino acid.

The nanoparticles in the present invention have a volume average particle diameter of 1 nm to 900 nm, but are preferably 10 nm to 700 nm, preferably 20 nm to 700 nm, from the viewpoint of absorption of the nanoparticles into the living body and retention control in the living body. It is preferable that it is 500 nm.
The volume average particle size of the nanoparticles can be measured by a method known in the art, for example, a commercially available wet particle size measuring device (for example, Zetasizer 3000HS _A manufactured by Malvern).

(Delivery object)
Examples of deliverables contained in the nanoparticles of the present invention include physiologically active substances and diagnostic agents. Examples of the biological activity of the physiologically active substance include therapeutic and prophylactic activities. These deliverables can be used singly or in combination of two or more.

  Examples of the physiologically active substance include inorganic compounds, organic compounds, polypeptides, nucleic acids, polysaccharides and the like having therapeutic, diagnostic and / or prophylactic activities. In addition, biological activities of physiologically active substances include vascular action, neuroactive action, hormonal action, anticoagulant action, immunomodulatory action, anti-inflammatory analgesic action, antibacterial action, antitumor action, antiviral action, antigenic action, Examples thereof include, but are not limited to, antibody action and antisense activity.

  “Polypeptide” as used herein means any size comprising at least two or more amino acids, with or without post-translational modifications such as glycosylation or phosphorylation. Examples of polypeptides include insulin, calcitonin, immunoglobulin, antibody, cytokine (eg, lymphokine, monokine, chemokine), interleukin, interferon, erythropoietin, nuclease, tumor necrosis factor, colony stimulating factor, enzyme (eg, superoxide Dismutase, tissue plasminogen activator), tumor suppressor, blood proteins, hormones and hormone analogs (eg, growth hormone, corticotropin, luteinizing hormone releasing hormone), vaccine (eg, tumor antigen, bacterial antigen, viral antigen) And complete proteins such as antigens, blood coagulation factors, growth factors, granulocyte colony-stimulating factors, muteins and active fragments thereof. Examples of the physiologically active action of the polypeptide include, but are not limited to, protein inhibitors, protein antagonists, and protein agonists. As used herein, “nucleic acid” refers to DNA or RNA sequences of any length, including genes (eg, antisense molecules that can bind to complementary DNA to inhibit transcription) and ribozymes. Can be mentioned. Polysaccharides such as heparin can also be used.

In addition, the nanoparticle of the present invention can contain a physiologically active substance for detecting an analyte for diagnosis. For example, an antigen, an antibody (monoclonal or polyclonal), a receptor, a hapten, an enzyme, a protein, a polypeptide, a nucleic acid (eg, DNA or RNA), a hormone, or a polymer can be mentioned, and at least one of these can be converted into nanoparticles. It can be included. In addition, the nanoparticles themselves can be labeled so that the nanoparticles can be easily detected. Examples of labels can include, but are not limited to, various enzymes, fluorescent materials, luminescent materials, bioluminescent materials and radioactive materials. Examples of suitable enzymes can include horseradish peroxidase, alkaline phosphatase, β-galactosidase, and acetylcholinesterase. Examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride and phycoerythrin. An example of the luminescent material is luminol. Examples of bioluminescent materials include luciferase, luciferin and aequorin. Examples of suitable radioactive materials include ¹²⁵ I, ¹³¹ I, ³⁵ S, and ³ H.

  Particularly useful physiologically active substances include various hormones (for example, insulin, estradiol, etc.), asthma (for example, albuterol), tuberculosis (for example, rifampicin) in terms of systemic and local treatments. , Ethambutol, streptomycin, isoniazid, pyrazinamide, etc.), cancer drugs (eg cisplatin, carboplatin, adriamycin, 5-FU, paclitaxel etc.) and hypertension drugs (eg clonidine, prazosin, propranolol, labetalol, bunitrolol) , Reserpine, nifedipine, furosemide, etc.), but is not limited thereto.

  The nanoparticles can include any diagnostic agent. By administering the nanocomposite particles to the patient, the diagnostic agent can be delivered locally or systemically. Specific examples of the diagnostic agent include a contrast agent, but are not limited thereto. As contrast agents, commercially available positron tomography (PET), computer linked tomography (CT), single photon emission computed tomography, X-ray, fluoroscopy, magnetic resonance imaging (MRI) Drugs can be mentioned. Diagnostic agents can be detected using standard techniques available in the art and commercially available equipment.

  From the viewpoint of stability of the nanoparticles and the delivery target, the nanoparticles contain 1% by mass to 50% by mass, preferably 5% by mass to 30% by mass, of the drug based on the total mass of the nanoparticles. Can do. The content of the drug can be appropriately selected depending on the desired effect, the release rate of the delivery target or the release period.

(Biodegradable polymer)
The nanoparticles in the present invention include at least one biodegradable polymer. In general, nanoparticles formed containing a biodegradable polymer are decomposed into substances that are harmless to the living body by enzymatic degradation or by contact with water in the living body. Examples of the biodegradable polymer include, but are not limited to, polyester compounds, polyamide compounds, polycarbonate compounds, polyvinyl compounds, and polysaccharides. These biodegradable polymers can be used singly or in combination of two or more.

Specific examples of the polyester compound include polylactic acid, polyglycolic acid, polycaproic acid and copolymers thereof (for example, poly (lactic acid-glycolic acid) copolymer), polybutylene succinate, polyethylene succinate, Poly (butylene succinate / adipate), poly (butylene succinate / carbonate), poly (butylene succinate / terephthalate), poly (butylene adipate / terephthalate), poly (tetramethylene adipate / terephthalate) and poly (butylene succinate / Adipate / terephthalate), but is not limited thereto.
Specific examples of the polyamide compound may include polyleucine, specific examples of the polyvinyl compound may include polycyanoacrylate, and specific examples of the polysaccharide may include cellulose. It is not limited to.

  Among these biodegradable polymers, polylactic acid, polyglycolic acid, poly (lactic acid-glycolic acid) (PLGA), and poly (polyglycolic acid) from the viewpoints of biocompatibility, biodegradability, stability in aqueous dispersion, etc. It is preferable to use at least one selected from cyanoacrylates, it is more preferable to use at least one selected from polylactic acid, polyglycolic acid, and PLGA, and it is more preferable to use PLGA.

By appropriately selecting the type and molecular weight of the biodegradable polymer, the release rate and biodegradation rate of the drug from the nanoparticles can be controlled. For example, when the biodegradable polymer is PLGA, the preferred molecular weight is 1500-150,000, more preferably 1500-75000. Thereby, the nanoparticle which is easy to release a physiologically active substance can be provided. When the biodegradable polymer is a copolymer, the drug release rate and biodegradation rate from the nanoparticles can be controlled by appropriately selecting the composition ratio of each monomer.
Furthermore, the nanoparticles may be modified so that the release rate and biodegradation rate of the drug from the nanoparticles can be controlled.

  Although there is no restriction | limiting in particular as content of the biodegradable polymer in the nanoparticle of this invention, For example, it can be 50-99 mass% with respect to the total mass of a nanoparticle, and it should be 70-95 mass% Is preferred.

(amino acid)
The nanoparticles of the present invention contain at least one amino acid. By including an amino acid, the dispersion stability of the nanoparticles is further improved, and the redispersibility of the nanoparticles is further improved.
For example, this can be considered as follows. In other words, it is thought that the dispersion stability of nanoparticles is improved by the attachment of amino acids to the surface of nanoparticles containing the delivery target and the biodegradable polymer due to electrostatic repulsion between nanoparticles derived from amino acids. it can.

  The amino acid in the present invention is not particularly limited as long as it is a compound containing at least one amino group and at least one carboxyl group in one molecule, and α-amino acids such as β-amino acids may be α-amino acids. In addition, when an optical isomer exists, any of L-form, D-form, and racemate may be used.

  Specific examples of amino acids in the present invention include aspartic acid, glutamic acid, lysine, arginine, histidine, glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, proline, phenylalanine, tyrosine, and Examples include naturally-derived α-amino acids such as tryptophan and optical isomers thereof, β-alanine, sarcosine, ornithine, creatine, γ-aminobutyric acid, opine, and the like.

Among these, from the viewpoint of nanoparticle redispersibility, a basic amino acid containing at least two basic groups is preferable, and a basic amino acid selected from lysine, arginine, and histidine is more preferable.
In the present invention, amino acids may be used alone or in combination of two or more.

Although there is no restriction | limiting in particular as content of the amino acid in the nanoparticle of this invention, For example, it can be 0.1-20 mass% with respect to the total mass of a nanoparticle, and is 0.5-15 mass% It is preferable.
The amino acid content can be appropriately selected according to the type of amino acid. For example, when a basic amino acid is used as the amino acid, it can be 0.1 to 20% by mass with respect to the total mass of the nanoparticles, and is 0.5 to 15% by mass from the viewpoint of redispersibility. preferable.
For example, when using a neutral amino acid or acidic amino acid as an amino acid, it can be 0.1-20 mass% with respect to the total mass of a nanoparticle, and 0.5-15 mass% from a redispersibility viewpoint. It is preferable that

Moreover, the said nanoparticle may further contain various additives as needed. Specific examples of the additive include, but are not limited to, a surfactant, a water-soluble polymer, and a lipid. By containing an additive, the redispersibility of the nanoparticles can be further improved.
Specific examples of the water-soluble polymer include polyvinyl alcohol, polyethylene glycol, and polyethylene oxide, and specific examples of the lipid include phospholipid and cholesterol. These additives can be used alone or in combination of two or more.

  In the nanoparticle of the present invention, from the viewpoint of redispersibility, the biodegradable polymer contains at least one selected from polylactic acid, polyglycolic acid and PLGA, and the content of the delivery target is the total mass of the nanoparticle. 1 to 50% by mass with respect to the total mass of the nanoparticles, and preferably 0.1 to 20% by mass of the α-amino acid. Including at least one selected from glycolic acid and PLGA, the content of the delivery target is 1 to 50% by mass with respect to the total mass of the nanoparticles, and at least one basic amino acid is added to the total mass of the nanoparticles It is more preferable to contain 0.1-20 mass% with respect to.

  The nanoparticle of the present invention can be obtained as a nanoparticle dispersion by, for example, bringing nanocomposite particles produced by the method for producing nanocomposite particles described later into contact with water.

<Nanocomposite particles>
The nanocomposite particles of the present invention contain at least one kind of the above-mentioned nanoparticles and have a volume average particle diameter of 0.5 μm or more and 500 μm or less. With such a configuration, the nano-particle composite particles are excellent in redispersibility when brought into contact with water.
The nanocomposite particles may be composed of two or more of the nanoparticles, or may further include other components (preferably saccharides) in addition to the nanoparticles. The nanocomposite particles composed of two or more of the nanoparticles are, for example, those in which two or more of the nanoparticles form an aggregate composed of nanoparticles by intermolecular force.

In the present invention, the volume average particle size of the nanocomposite particles is preferably 0.5 to 500 μm, more preferably 1 to 200 μm. Note that the volume average particle size of the nanocomposite particles is a dry particle size measuring device. (For example, LDSA3500A manufactured by Tohnichi Computer Applications Co., Ltd.) is used to measure by a conventional method.

  The details of the nanoparticles constituting the nanocomposite particles of the present invention are as described above, and the preferred embodiments are also the same.

The nanocomposite particles may further contain at least one water-soluble compound in addition to the nanoparticles. The water-soluble compound is not particularly limited, but is preferably at least one selected from saccharides from the viewpoint of redispersibility.
In addition, the water-soluble compound in the present invention means a compound having a solubility in 100 g of pure water at 25 ° C. of 10 g or more.

Examples of the saccharide include monosaccharides, disaccharides, oligosaccharides, polysaccharides, and sugar alcohols. Among these, from the viewpoint of redispersibility, at least one selected from disaccharides and sugar alcohols is preferable.
Specific examples include glucose, sorbitol, mannitol, sucrose, maltose, lactose, trehalose, dextran, cyclodextrin, amylose, agarose, hyaluronic acid and the like, but are not limited thereto. Among these sugar substances, sorbitol, mannitol, sucrose, maltose, lactose, and trehalose are preferably used, and trehalose and lactose are more preferably used from the viewpoint of the water solubility of the sugar substance and the redispersibility of the nanoparticles. . Moreover, these saccharides can be used individually by 1 type or in mixture of 2 or more types.

  There is no restriction | limiting in particular as a content ratio of the nanoparticle and water-soluble compound in this invention, For example, it is set as 10: 1 to 1:10 as mass ratio (nanoparticle: water-soluble compound) of the nanoparticle with respect to a water-soluble compound. be able to. Among these, from the viewpoint of nanoparticle redispersibility and production suitability, the ratio is preferably 10: 1 to 1: 5, and more preferably 10: 1 to 1: 1.

  Moreover, the nanocomposite particles of the present invention may contain various additives as necessary. Examples of additives include buffer salts, fatty acids, fatty acid esters, phospholipids, inorganic compounds, phosphates and the like.

<Method for producing nanocomposite particles>
The method for producing nanocomposite particles of the present invention comprises mixing a solution containing at least one kind of a delivery object, at least one kind of biodegradable polymer, and at least one kind of an organic solvent with water, And a dispersion step of obtaining a nanoparticle dispersion containing a biodegradable polymer, and a drying step of removing water from the nanoparticle dispersion in the presence of an amino acid, and other steps as necessary. Consists of including.
By removing water from a dispersion containing a delivery target and a biodegradable polymer in the presence of an amino acid, nanocomposite particles containing the delivery target and the biodegradable polymer and having excellent redispersibility can be efficiently obtained. Can be manufactured.

(Dispersion process)
In the dispersion step, a solution containing a delivery target, a biodegradable polymer, and an organic solvent is mixed with water to prepare a dispersion of nanoparticles containing the delivery target and the biodegradable polymer.
A delivery object and a biodegradable polymer are synonymous with the delivery object and biodegradable polymer in the above-mentioned nanoparticle, and a preferable aspect is also the same.

  The organic solvent in the present invention is not particularly limited as long as it can form a solution together with the delivery target and the biodegradable polymer, and may be a hydrophilic organic solvent or a hydrophobic organic solvent.

  Specific examples of the hydrophilic organic solvent include alcohol solvents such as ethanol, isopropyl alcohol, ethylene glycol, and glycerin; ketone solvents such as acetone and methyl ethyl ketone; tetrahydrofuran, dioxane, diethylene glycol, triethylene glycol, diethylene glycol monoethyl ether, and the like. And ether solvents such as: amide solvents such as dimethylformamide, dimethylacetamide and dimethylimidazolidinone; sulfoxide solvents such as dimethyl sulfoxide.

  Specific examples of the hydrophobic organic solvent include aliphatic solvents such as hexane and heptane, halogen solvents such as dichloromethane, chloroform and chlorobenzene, aromatic solvents such as toluene and xylene, and ketone solvents such as cyclohexanone. And ester solvents such as ethyl acetate and butyl acetate.

In the present invention, when a hydrophilic organic solvent is used, it is preferably at least one selected from an alcohol solvent, a ketone solvent, and an ether solvent from the viewpoint of redispersibility and safety. More preferably, it is at least one selected from ether solvents.
By preparing a nanoparticle dispersion using a hydrophilic organic solvent, a safer nanoparticle dispersion can be efficiently prepared.

  As a method for preparing a solution containing a delivery target, a biodegradable polymer, and an organic solvent, a solution preparing method that is usually performed can be applied without particular limitation. For example, the biodegradable polymer and the delivery target can be added to an organic solvent and stirred.

  In addition, as a method of mixing a delivery target, a biodegradable polymer, and a solution containing an organic solvent and water, a liquid mixing method that is usually performed can be applied without any particular limitation. For example, it can be performed using a general stirring device, a homogenizer, an ultrasonic generator, or the like.

Further, the mixing method of the solution and water may be a method of adding water to the solution or a method of adding the solution to water. From the viewpoint of production efficiency, the solution is added to water. The method of adding is preferable.
Moreover, there is no restriction | limiting in particular about the speed | rate which mixes the said solution and water. In the present invention, a dispersion of nanoparticles having a narrow particle size distribution can be obtained by mixing the solution and water at a constant mixing speed.

In the present invention, the concentration of the solution mixed with water is not particularly limited. For example, the total content of the delivery target and the biodegradable polymer is 0.1 to 10% by mass with respect to the total solution mass. It is preferably 0.5 to 3% by mass.
The mixing ratio of the solution to water (solution: water) is preferably 45:55 to 1: 99% by mass, and more preferably 40:60 to 5: 95% by mass.

  In the present invention, as a biodegradable polymer, at least one selected from polylactic acid, polyglycolic acid, PLGA, and polycyanoacrylate is used, whereby a nanoparticle dispersion containing a delivery target and a biodegradable polymer is obtained. It can be prepared more efficiently. This can be considered, for example, because these biodegradable polymers have self-dispersibility.

(Drying process)
In the drying step, water is removed from the nanoparticle dispersion in the presence of amino acids. The presence of amino acids when water is removed from the nanoparticle dispersion improves the redispersibility of the nanocomposite particles formed after the water removal with respect to water.
As an amino acid in a drying process, it is synonymous with the amino acid in the above-mentioned nanoparticle, and its preferable aspect is also the same.

  In the present invention, it is sufficient that an amino acid is present when water is removed from the dispersion of nanoparticles, and the timing of adding the amino acid is not limited. For example, in the step of obtaining the nanoparticle dispersion, an amino acid may be added to water mixed with the solution, or an amino acid may be added after mixing the solution and water. Further, an amino acid may be added immediately before removing water in the drying step.

In the present invention, a step of removing the organic solvent may be further provided before removing water from the nanoparticle dispersion. The method for removing the organic solvent is not particularly limited, and examples thereof include a method of stirring the nanoparticle dispersion, a method of bubbling an inert gas such as nitrogen gas, and air.
Furthermore, in this invention, you may remove an organic solvent with water. Thereby, nanocomposite particles can be manufactured more efficiently.

The method for removing water from the nanoparticle dispersion is not particularly limited, but from the viewpoint of the stability of the delivery target, either the spray drying method or the freeze drying method is preferable.
The lyophilization method can be carried out by a conventional method using a commonly used lyophilization apparatus.

  In addition, as a spray drying method, for example, general spray drying techniques described in “Spray Drying Handbook” by K. Masters, John Wiley & Sons, New York, 1984 are particularly limited. Can be used. In general, in the spray drying method, by using heat derived from a hot gas such as heated air or heated nitrogen, from the droplets formed by the atomizer, the inlet temperature immediately after the atomization and the outlet temperature at the end of drying Under the following temperature gradient, the solvent can be evaporated to obtain spray-dried particles.

  As the atomization apparatus, a spray technique known in the art can be used. For example, a hydraulic press nozzle atomizing device, a two-fluid atomizing device, a sonic atomizing device, a centrifugal disk atomizing device including a rotating disk or wheels can be used, but is not limited thereto.

In the method for producing nanocomposite particles of the present invention, the nanoparticle dispersion may further contain at least one saccharide. Thereby, the nanocomposite particle containing a nanoparticle and saccharides can be manufactured. When the nanocomposite particles further contain a saccharide, nanocomposite particles having excellent water redispersibility and a desired particle size can be produced more efficiently.
The saccharide in the present invention is synonymous with the saccharide in the nanocomposite particles described above, and the preferred embodiment is also the same.
The mass ratio of saccharides contained in the dispersion of nanoparticles can be 10: 1 to 1:10 as nanoparticles: saccharides. From the viewpoint of nanoparticle redispersibility, 10: 1 to 1: 5 is preferable, and 10: 1 to 1: 1 is more preferable.

  In the method for producing nanocomposite particles of the present invention, a dispersion of nanoparticles can be rapidly prepared by mixing an organic solvent solution containing a delivery target and a biodegradable polymer with water. Furthermore, since nanocomposite particles having excellent redispersibility can be prepared by removing water from the nanoparticle dispersion in the presence of amino acids, it is extremely efficient as a method for producing nanocomposite particles.

  EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples. Unless otherwise specified, “part” and “%” are based on mass.

Example 1
Rifampicin (RFP, manufactured by Sigma Aldrich) 0.1 g as a delivery target, and poly (lactic acid-glycolic acid) (PLGA, lactic acid / glycolic acid = 75/25, weight average molecular weight 10,000, Wako Pure Chemical as biodegradable polymer A solution in which 0.9 g) was dissolved in 10 ml of dichloromethane was prepared.
In 100 ml of purified water, 0.15 g of arginine (manufactured by Wako Pure Chemical Industries, Ltd.) as an amino acid was dissolved, and this and the solution obtained above were mixed using a probe type sonicator (manufactured by Branson), and then 200 rpm at room temperature. The mixture was stirred at a stirring speed for 12 hours to obtain a dispersion of PLGA nanoparticles containing rifampicin.
The resulting dispersion of nanoparticles, measurement of the volume average particle diameter using Zetasizer 3000HS _A (produced by Malvern Co.) was 62.7Nm.

The nanoparticle dispersion obtained above was supplied to a spray drying apparatus (manufactured by BUCHI, mini spray dryer B290 type), spray-dried, and freeze-dried overnight to prepare nanocomposite particles. The spray drying conditions were as follows: the spray gun diameter used was 0.75 mm, the inlet temperature was 68 ° C., the dry air amount was 22 m ³ / hr, the spray air amount was 500 L / hr, and the sample supply rate was 1.25 mL / min. .
It was 2.83 micrometers when the volume average particle diameter of the obtained nanocomposite particle | grains was measured using the dry-type particle diameter measuring apparatus (The Tohnichi Computer Applications company make, LDSA-3500A).

When purified water was added to the obtained nanocomposite particles, a nanoparticle dispersion was immediately obtained. The volume average particle diameter of the redispersed nanoparticles was measured in the same manner as described above, and found to be 64.8 nm.
Further, the obtained nanocomposite particles were stored at −30 ° C. for 1 month, and then purified water was added. As a result, a nanoparticle dispersion was immediately obtained. The volume average particle diameter of the redispersed nanoparticles was measured in the same manner as described above, and it was 63.3 nm. Furthermore, after storing for 6 months, it was 62.7 nm.

(Example 2)
In Example 1, nanoparticles and nanocomposite particles were prepared in the same manner as in Example 1 except that the amount of arginine added was changed to 2.0 g.
The obtained nanoparticles had a volume average particle diameter of 78 nm.
The volume average particle diameter of the nanoparticles redispersed in the same manner as in Example 1 was 88 nm.

(Example 3)
In Example 1, nanoparticles and nanocomposite particles were prepared in the same manner as in Example 1 except that glutamine was used instead of arginine and the addition amount was 2.0 g.
The volume average particle diameter of the obtained nanoparticles was 234.6 nm.

Example 4
In Example 1, nanoparticles and nanocomposite particles were prepared in the same manner as in Example 1 except that glutamic acid was used instead of arginine and the addition amount was changed to 0.5 g.
The volume average particle diameter of the obtained nanoparticles was 433.2 nm.

(Example 5)
In Example 1, nanoparticles were prepared in the same manner as in Example 1 except that lysine was used instead of arginine and the addition amount was changed to 2.0 g.
The volume average particle diameter of the obtained nanoparticles was 89.6 nm.

(Example 6)
In Example 1, nanoparticles and nanocomposite particles were prepared in the same manner as in Example 1 except that histidine was used instead of arginine and the addition amount was changed to 2.0 g.
The volume average particle diameter of the obtained nanoparticles was 87.2 nm.

(Example 7)
Rifampicin (RFP, manufactured by Sigma-Aldrich) 0.05 g as a delivery target, and poly (lactic acid-glycolic acid) (PLGA, lactic acid / glycolic acid = 75/25, weight average molecular weight 10,000, Wako Pure Chemical as biodegradable polymer A solution prepared by dissolving 0.45 g (manufactured by Kogyo) in 10 ml of acetone was prepared.
In 100 ml of purified water, 0.15 g of arginine (manufactured by Wako Pure Chemical Industries, Ltd.) as an amino acid was dissolved, and this was mixed with the solution obtained above, and then stirred at room temperature for 15 hours at a stirring speed of 200 rpm. A dispersion of PLGA nanoparticles containing was obtained.
With respect to the obtained dispersion of nanoparticles, the volume average particle diameter was measured in the same manner as described above, and it was 163.8 nm.
The nanoparticle dispersion obtained above was spray-dried in the same manner as described above to prepare nanocomposite particles. The volume average particle diameter of the obtained nanocomposite particles was measured in the same manner as described above. It was 50 μm.
When purified water was added to the obtained nanocomposite particles, a nanoparticle dispersion was immediately obtained. The volume average particle diameter of the redispersed nanoparticles was measured in the same manner as described above, and was 163.8 nm.

(Example 8)
Rifampicin (RFP, manufactured by Sigma Aldrich) 0.1 g as a delivery target, and poly (lactic acid-glycolic acid) (PLGA, lactic acid / glycolic acid = 75/25, weight average molecular weight 10,000, Wako Pure Chemical as biodegradable polymer A solution in which 0.9 g) was dissolved in 10 ml of dichloromethane was prepared.
After dissolving 0.15 g of arginine (manufactured by Wako Pure Chemical Industries, Ltd.) as an amino acid in 100 ml of purified water and mixing this and the solution obtained above using a probe sonicator (manufactured by Branson), PLGA containing rifampicin A dispersion of nanoparticles was obtained.

The nanoparticle dispersion obtained above was supplied to a spray drying apparatus (manufactured by BUCHI, mini spray dryer B290 type), spray-dried, and freeze-dried overnight to prepare nanocomposite particles. The spray drying conditions were as follows: the spray gun diameter used was 0.75 mm, the inlet temperature was 68 ° C., the dry air amount was 22 m ³ / hr, the spray air amount was 500 L / hr, and the sample supply rate was 1.25 mL / min. .
It was 1.94 micrometers when the volume average particle diameter of the obtained nanocomposite particle | grains was measured using the dry-type particle diameter measuring apparatus (The Tohnichi Computer Applications company make, LDSA-3500A).

  When purified water was added to the obtained nanocomposite particles, a nanoparticle dispersion was immediately obtained. The volume average particle diameter of the redispersed nanoparticles was measured in the same manner as described above, and it was 59.9 nm.

Example 9
In Example 8, 0.4 g of poly (lactic acid-glycolic acid) (PLGA, lactic acid / glycolic acid = 75/25, weight average molecular weight 20000, manufactured by Wako Pure Chemical Industries, Ltd.) was used as a biodegradable polymer, and the amount of algin used A nanoparticle dispersion and nanocomposite particles were prepared in the same manner as in Example 6 except that the amount was 0.1 g.
The obtained nanocomposite particles had a volume average particle size of 5.73 μm. Moreover, when this was re-dispersed in purified water, the volume average particle diameter of the obtained nanoparticle dispersion was 100.9 nm.

(Example 10)
Rifampicin (RFP, manufactured by Sigma Aldrich) 0.1 g as a delivery target, and poly (lactic acid-glycolic acid) (PLGA, lactic acid / glycolic acid = 75/25, weight average molecular weight 10,000, Wako Pure Chemical as biodegradable polymer A solution prepared by dissolving 0.9 g in 10 ml of acetone was prepared.
In 100 ml of purified water, 0.15 g of arginine (manufactured by Wako Pure Chemical Industries, Ltd.) as an amino acid was dissolved, and this was mixed with the solution obtained above, and then stirred at room temperature for 15 hours at a stirring speed of 200 rpm. A dispersion of PLGA nanoparticles containing was obtained.
With respect to the obtained dispersion of nanoparticles, the volume average particle diameter was measured using a Zetasizer 3000HS _A (manufactured by Malvern), and it was 100.5 nm.

The nanoparticle dispersion obtained above was freeze-dried to prepare nanocomposite particles.
When purified water was added to the obtained nanocomposite particles, a nanoparticle dispersion was immediately obtained. The volume average particle size of the redispersed nanoparticles was measured in the same manner as described above, and found to be 106.5 nm.

(Example 11)
Rifampicin (RFP, manufactured by Sigma Aldrich) 0.1 g as a delivery target, and poly (lactic acid-glycolic acid) (PLGA, lactic acid / glycolic acid = 75/25, weight average molecular weight 10,000, Wako Pure Chemical as biodegradable polymer A solution in which 0.9 g) was dissolved in 10 ml of dichloromethane was prepared.
In 100 ml of purified water, 0.15 g of arginine (manufactured by Wako Pure Chemical Industries, Ltd.) as an amino acid was dissolved, and this and the solution obtained above were mixed using a probe type sonicator (manufactured by Branson), and then 200 rpm at room temperature. The mixture was stirred at a stirring speed for 12 hours to obtain a dispersion of PLGA nanoparticles containing rifampicin.
When the volume average particle diameter of the obtained dispersion of nanoparticles was measured using a Zetasizer 3000HS _A (manufactured by Malvern), it was 60.2 to 62.7 nm.

After adding 0.85 g each of mannitol, lactose and trehalose as sugars to the nanoparticle dispersion obtained above and dissolving them, they are supplied to a spray dryer (BUCHI, mini spray dryer B290 type) and sprayed. The nanocomposite particles were prepared by drying and then freeze-drying overnight. The spray drying conditions were as follows: the spray gun diameter used was 0.75 mm, the inlet temperature was 68 ° C., the dry air amount was 22 m ³ / hr, the spray air amount was 500 L / hr, and the sample supply rate was 1.25 mL / min. .
When the volume average particle size of the obtained nanocomposite particles was measured using a dry particle size measuring device (LDSA-3500A, manufactured by Tohnichi Computer Applications), 2.08 μm for mannitol, 1.51 μm for lactose, and 1.51 μm for trehalose. It was 1.51 μm.

  When purified water was added to the obtained nanocomposite particles, a nanoparticle dispersion was immediately obtained. The volume average particle diameter of the redispersed nanoparticles was measured in the same manner as described above, and it was found to be 72.3 nm for mannitol, 68.4 nm for lactose, and 66.8 nm for trehalose.

(Example 12)
Rifampicin (RFP, manufactured by Sigma Aldrich) 0.1 g as a delivery target, and poly (lactic acid-glycolic acid) (PLGA, lactic acid / glycolic acid = 75/25, weight average molecular weight 10,000, Wako Pure Chemical as biodegradable polymer A solution in which 0.4 g) was dissolved in 10 ml of dichloromethane was prepared.
In 100 ml of purified water, 0.05 g of arginine (manufactured by Wako Pure Chemical Industries, Ltd.) and 0.15 g of cysteine (manufactured by Wako Pure Chemical Industries, Ltd.) were dissolved as amino acids, and the resulting solution was used as a probe sonicator (manufactured by Branson). Then, a dispersion of PLGA nanoparticles containing rifampicin was obtained.

The nanoparticle dispersion obtained above was spray-dried in the same manner as described above to prepare nanocomposite particles.
It was 1.33 micrometer when the volume average particle diameter of the obtained nanocomposite particle was measured similarly to the above.

  When purified water was added to the obtained nanocomposite particles, a nanoparticle dispersion was immediately obtained. The volume average particle diameter of the redispersed nanoparticles was measured in the same manner as described above, and found to be 138.7 nm.

  Furthermore, when the encapsulation rate of rifampicin was measured using HPLC for the obtained nanocomposite particles, the encapsulation rate was 7.74% (theoretical value 20%), and the encapsulation efficiency was 38.7%. High inclusion efficiency.

  From the above, it can be seen that the nanocomposite particles of the present invention are excellent in nanoparticle redispersibility. Moreover, it turns out that the manufacturing method of the nanocomposite particle | grains of this invention is simple and efficient.

Claims (7)

  A nanoparticle comprising a delivery target, a biodegradable polymer, and an amino acid, and having a volume average particle diameter of 1 nm to 900 nm.   The nanoparticle according to claim 1, wherein the amino acid is a basic amino acid.   The nanoparticle according to claim 1 or 2, wherein the biodegradable polymer includes at least one selected from polylactic acid, polyglycolic acid, poly (lactic acid-glycolic acid), and polycyanoacrylate.   The nanoparticle according to any one of claims 1 to 3, wherein the volume average particle diameter is 10 nm or more and 700 nm or less.   Nanocomposite particles comprising at least one kind of nanoparticles according to any one of claims 1 to 4 and having a volume average particle diameter of 1 µm or more and 500 µm or less.   Furthermore, the nanocomposite particle of Claim 5 containing at least 1 sort (s) of saccharides. A dispersion step of mixing a solution containing a delivery target, a biodegradable polymer, and an organic solvent with water to obtain a dispersion of nanoparticles containing the delivery target and the biodegradable polymer;
A drying step of removing water from the dispersion of nanoparticles in the presence of an amino acid;
A method for producing nanocomposite particles comprising: