Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
  • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
  • A61K9/51—Nanocapsules; Nanoparticles
  • A61K9/5107—Excipients; Inactive ingredients
  • A61K9/5123—Organic compounds, e.g. fats, sugars
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
  • A61K31/542—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with heterocyclic ring systems
  • A61K31/545—Compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins, cefaclor, or cephalexine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/66—Phosphorus compounds
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
  • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
  • A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
  • A61K31/727—Heparin; Heparan
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/18—Growth factors; Growth regulators
  • A61K38/1816—Erythropoietin [EPO]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/19—Cytokines; Lymphokines; Interferons
  • A61K38/21—Interferons [IFN]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/22—Hormones
  • A61K38/23—Calcitonins
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/22—Hormones
  • A61K38/27—Growth hormone [GH], i.e. somatotropin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/22—Hormones
  • A61K38/28—Insulins
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/22—Hormones
  • A61K38/31—Somatostatins
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
  • A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
  • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/08—Solutions
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
  • A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
  • A61K9/146—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic macromolecular compounds
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
  • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
  • A61K9/51—Nanocapsules; Nanoparticles
  • A61K9/5107—Excipients; Inactive ingredients
  • A61K9/513—Organic macromolecular compounds; Dendrimers
  • A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
  • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
  • A61K9/51—Nanocapsules; Nanoparticles
  • A61K9/5192—Processes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
  • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B82—NANOTECHNOLOGY
  • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
  • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Definitions

• the present invention relates to orally administrable nanoparticle compositions
• the present invention relates to orally
• nanoparticles are prepared by binding water-soluble drugs with
• Water-soluble drugs including physiological active agents, protein drugs, have low
• intravenous injection has been usually used for the drugs.
• intravenous injection has been usually used for the drugs.
• w/o emulsions may be usually disrupted by phase transition upon administration into the body, and thus, drugs
• the particle size of the prepared emulsion is extremely large, 10 to 20 micrometers
• the emulsion has low drug entrapping
• Korean Patent Laid-open No. 2002-66776 discloses orally administrable
• the carrier consists of only lipid, and thus, may be degraded by lipases in vivo.
• micrometer-sized particles cannot be absorbed into the intestinal tract, and are generally
• 5 ⁇ m-sized PLGA microspheres have only low insulin entrapping rate and deliver insulin less effectively than nanometer-sized particles.
• United States Patent 5,962,024 discloses dissolving drugs at pH 6.5 or
• microspheres formed only from enteric polymers have the
• polymers are dissolved in the intestinal tract and the drugs entrapped in the microspheres
• WO 94/08599 discloses preparation of complexes wherein insulin is ionically
• This preparation method is to dissolve insulin-sodium
• United States Patent 5,858,410 discloses nano-particulating water-insoluble drugs by milling and using a microfluidizer, but this has the drawback that this is not suitable for water-soluble drugs unstable in vivo, e.g. protein drugs, because the drugs are directly exposed to in vivo environment.
• lipid nanoparticles should contain appropriate amounts of
• the present inventors have repeated extensive studies to solve the above-described problems and to develop drag carriers satisfying the above technical requirements. As a result, the present inventors discovered that drug entrapping rate and resistance against degradation enzymes in vivo can be increased by preparing orally administrable nanoparticle compositions with the particle size of 500 nm or less from water-soluble drag-counter-ion substance complexes and a certain ratio of a lipid and a polymer, etc., and completed the present invention.
• the purpose of the present invention is to provide orally administrable
• nanoparticle compositions with enhanced drug entrapping rate and resistance against
• lipases by containing complexes of water-soluble drugs and counter-ion substances thereby
• Fig. 1 is a graph showing stability of the orally administrable nanoparticle
• composition of the present invention containing insulin as an active ingredient against
• pancreatin prote/lipase
• Fig. 2 is a graph showing the relative blood glucose concentration after
• Fig. 3 is a graph showing the relative blood glucose concentration after
• Reference group 1 insulin-containing buffer of pH 7.4, dosage of insulin 20
• Example 1 (dosage of insulin 20 IU/kg)
• -A- Comparative Example 1 (dosage of insulin 20 IU/kg)
• Fig. 4 is a graph showing the relative blood glucose concentration after
• Reference group 2 insulin injectable solution, subcutaneous injection
• Fig. 5 is a graph showing the concentration of insulin after intra-intestinally
• Control group oral nanoparticle composition with no insulin
• Example 1 (dosage of insulin 20 IU/kg) - A-: Comparative Example 1 (dosage of insulin 20 IU/kg) -X-: Example la (dosage of insulin 20 IU/kg)
• Fig. 6 is a graph showing the concentration of insulin after intra-intestinally
• Fig. 7 is a graph showing the concentration of ceftriaxone after intra-duodenally
• ceftriaxone as an active ingredient to normal rats in Experiment 4(1)1).
• - ⁇ - Example 6 (dosage of ceftriaxone 40 mg/kg)
• Fig. 8 is a graph showing the concentration of ceftriaxone after intra-duodenally
• ceftriaxone as an active ingredient to normal rats in Experiment 4(1)1).
• - ⁇ - Example 6 (dosage of ceftriaxone 40 mg/kg)
• Reference group 2 ceftriaxone injectable solution, intravenous injection
• the present invention provides an orally administrable composition containing
• water-soluble drug is bonded with the counter-ion substance, 0.5 to 80 weight% of a lipid,
• lipid and said polymer is preferably in the range of 1 :0.2 ⁇ 1.
• the present invention provides a method for preparing the orally administrable nanoparticle composition according to the present invention, comprising the steps of: (a) ionically bonding a charged water-soluble drag with a counter-ion substance to form a complex of the water-soluble drug and the counter-ion substance; (b) adding a lipid, a polymer, and a solubilizing agent to the obtained complex and dissolving the whole mixture, and then, introducing the obtained solution to an aqueous solution containing an emulsifier to obtain a homogeneous liquid phase; (c) eliminating the solubilizing agent from the mixture obtained in step (b); and (d) optionally, minimizing the particle size using a microfluidizer.
• the water-soluble drag is entrapped in the nanoparticle composition at the rate of 70% or more, and can be retained at the rate of 80%) or more in the presence of pancreatin enzymatic solution. Still further, the present invention provides a method for preparing the orally
• solubilizing agent from the mixture obtained in step (b); and (d) optionally, minimizing
• counter-ion substance 0.5 to 80 weight% of a lipid, 0.5 to 80 weight% of a polymer, and
• the weight ratio of said lipid and said polymer is preferably in the range of
• the complex of the water-soluble drag and the counter-ion substance is obtained by reaction of the charged drug with the counter-ion substance.
• the water-soluble drug is obtained by reaction of the charged drug with the counter-ion substance.
• used as an active ingredient may be ones charged in an aqueous solution, preferably, ones
• a negatively charged substance which can be ionically bonded to a positively charged substance
• charged drug may be preferably selected from the group consisting of sodium salt of C 8 ⁇ 18
• fatty acid such as sodium oleate, sodium lauryl sulfate, sodium caproate, sodium laurate,
• sodium salt of bile acid sodium alginate, and sodium carboxymethylcellulose.
• quaternary ammonium compound such as camitine salt
• the substance is preferably 0.1 to 30 weight% of the total weight of the nanoparticle
• the composition of the present invention shows high drag entrapping rate.
• the molar ratio of the water-soluble drag and the counter-ion substance in the complex of the water-soluble drug and the counter-ion substance may be adjusted depending on the number of ions of the water-soluble drag, preferably to 1 : ⁇ .1-20, more preferably to 1 :3 ⁇ 10.
• Said lipid may be preferably one or more selected from an aliphatic alcohol such as monoglyceride, diglyceride, propylene glycol ester of fatty acid (Capriol, etc.), glycerol ester of fatty acid (GELucier, etc.), cetostearyl alcohol, cetyl alcohol, etc., and may be preferably contained at 0.5 to 80 weight%, more preferably 0.5 to 30 weight%>, of the total weight of the nanoparticle composition.
• an aliphatic alcohol such as monoglyceride, diglyceride, propylene glycol ester of fatty acid (Capriol, etc.), glycerol ester of fatty acid (GELucier, etc.), cetostearyl alcohol, cetyl alcohol, etc.
• Said polymer may be preferably one or more selected from an enteric polymer
• chitosan hydroxypropyl methylcellulose and its derivative, ethylcellulose, methylcellulose,
• polyvinyl alcohols sodium alginate, and carbomer, and may be preferably contained at 0.5
• nanoparticle composition particularly, in case of using an enteric polymer, drugs labile
• gastric acid e.g. proteins
• Said polymer is insoluble in water or becomes viscous when dissolved in water,
• the size of the lipid In adding polymer to the lipid nanoparticle composition, the size of the lipid
• nanoparticle can be varied depending on content of the polymer, and the ratio of lipid and
• polymer should be appropriately adjusted to obtain the nanoparticle of 500 nm or less.
• Said emulsifier is preferably one or more selected from polyoxyethylene
• polyoxypropylene copolymer (commercial name: PoloxamerTM), polyethylene glycol alkyl
• LabrafilTM, LabrasolTM), glycerol fatty acid ester commercial name: PlurolTM oleique
• vitamin E polyethyleneglycol succinate (Vitamin E polyethylene glycol succinate), lecithin, sodium lauryl sulfate, and bile acid and derivatives thereof, and may be preferably
• nanoparticle composition contained at 1 to 80 weight% based on the total weight of the nanoparticle composition.
• the emulsifier may be contained at 30 to 80 weight% > .
• lipid/polymer nanoparticles are dispersed in an aqueous solution, the emulsifier stabilizes
• Said solubilizing agent may be preferably one or more selected from C ⁇ 8 alcohol,
• composition at 50 weight% or less.
• solubilizing agent alone cannot
• the above described emulsifier may be added thereto to obtain a
• composition may contain a conventionally used oral absorbability
• said oral absorbability enhancing agent at 0 to 20 weight%, and said oral absorbability enhancing agent may be
• p-glycoprotein inhibitor preferably selected from p-glycoprotein inhibitor, caraitine salt, chelating agent, etc.
• the present composition may be preferably lyophilized with 0.1-30 weight%>
• cryoprotective agent of a cryoprotective agent, and so be prepared to powder, and then to conventional oral
• the cryoprotective agent may be preferably a saccharide such as glucose, mannitol, sorbitol, trehalose, an amino acid such as arginine, a
• protein such as albumin, etc.
• the orally administrable nanoparticle composition of the present invention is
• solubilizing agent from the mixture obtained in step b); and (d) optionally, minimizing the
• the obtained complex of the water-soluble drag and the counter-ion substance is the obtained complex of the water-soluble drag and the counter-ion substance
• a pH adjusting agent may be used to confer charges on the protein drug.
• adjusting agent may be selected from an acidifying agent such as hydrochloric acid,
• an alkalifying agent such as sodium
• an acid, an alkali, and an emulsifying aid may be added thereto at 0.5-50
• the mixture may be heated, preferably to 30-70 ° C .
• step (c) if the solubilizing agent is highly volatile, it may be
• the solution is preferably cyclized 3 to 10 times at 100 MPa
• the particle size of the nanoparticle may be further reduced by heating.
• the heating temperature is preferably 30-70 °C .
• invention is one comprising the steps of: (a) ionically bonding a charged water-soluble resin
• solubilizing agent from the mixture obtained in step (b); and (d) optionally, minimizing the
• the obtained nanoparticle composition is liquid, and if desired, may be lyophilized
• cryoprotective agent to be obtained as powder.
• Said cryoprotective agent is used
• cryoprotective agent may be preferably used at 0.1-30 weight%> based on the total weight
• the cryoprotective agent may be preferably a
• saccharide such as lactose, mannitol, trehalose, etc.
• amino acid such as arginine, a
• protein such as albumin, etc.
• liquid and powder composition obtained as described above may be readily
• dispersion size forming the dispersion system is 500 nm or less depending on properties of used emulsifier or drug, which is much smaller than prior orally administrable formulations.
• the particle size of the present orally administrable nanoparticle composition is
• the nanoparticle powder composition of the present invention can be stored stable
• nanoparticle composition of the present invention can be formulated
• Example 1 Preparation of an orally administrable nanoparticle composition
• Insulin 50 mg was dissolved in 20 ml of citrate buffer of pH 3, and sodium docusate (Aldrich) dissolved in 80 ml of pH 3 citrate buffer was reacted therewith at the molar ratios of 1 :3, 1 :6 and 1 :9 to 1 mole of insulin, to obtain a complex of insulin and the counter-ion substance.
• insulin in the supernatant was analyzed according the insulin analysis method described in the US Pharmacopoeia, to obtain results as shown in the following Table 1.
• the complex prepared from insulin and the counter-ion substance at the molar ratio of 1 :6 was washed with each of citrate buffer of pH 3 and water three times, and then, used for the subsequent step.
• lipid/polymer nanoparticle 15 mg of Insulin-sodium docusate complex and 60 mg of mono-olein (TCI) were dissolved in 10 ml of ethanol.
• the solution was introduced into 20 ml of an aqueous solution containing 20 mg of chitosan (Korea Chitosan) and 200 mg of Poloxamer 407TM (BASF), and the whole mixture was stirred to obtain a homogenous solution, and then, ethanol was eliminated therefrom by injecting nitrogen gas thereto.
• 1% of citric acid was added thereto as an additive to increase the solubility of chitosan.
• the lipid/polymer nanoparticle was cyclized ten times at 100 MPa of room temperature using a
• EmulsiFlex-C5TM (Avestin)
• composition containing insulin containing insulin
• the particle size and degree of dispersion of the composition were measured according to photon spectrometry (ELS-8000TM, Otsuka electronics) after 200 ⁇ l of the obtained liquid composition was dispersed into 1 ml of distilled water.
• ELS-8000TM photon spectrometry
• the mean particle size of the liquid composition of Example 1 was 83.0 nm and the degree of dispersion thereof was 0.300.
• the liquid composition was prepared according to the same method as Example 1,
• the mean particle size was 117.4 nm
• the liquid composition was prepared according to the same method as Example 1,
• microfluidizer The particle size and degree of dispersion were measured according to the
• Example 2 the mean particle size was 241.8 nm and the degree of dispersion- was 0.090.
• Example 1 the mean particle size was 211.6 nm, and the degree of dispersion was 0.213.
• Example la The liquid composition was prepared according to the same method as Example 1 except that the microfluidizer was not used. As a result of measuring the particle size and degree of dispersion according to the method of Example 1 , the mean particle size was 234.3 nm, and the degree of dispersion was 0.291.
• Example 2 Preparation of an orally administrable nanoparticle powder
• the variation in the particle size could be minimized when the concentration of the cryoprotective agent was about 5%>.
• Example 3 Preparation 2 of an orally administrable nanoparticle formulation containing insulin
• Example 1 sodium lauryl sulfate (Sigma) was used at the ratio of 1:7 instead of sodium docusate to obtain a complex of insulin and the counter-ion substance.
• the complex of insulin and the counter-ion substance, mono-olein, and polyethyleneglycol 2000 were weighed 15 mg, 45 mg, and 90 mg, respectively, and were dissolved in 10 ml of ethanol. According to the same procedure, the orally administrable nanoparticle composition containing insulin was prepared.
• the particle size and degree of dispersion were measured, and as a result, the mean particle size was 121 nm, and the degree of dispersion was 0.314.
• Example 4 Preparation 3 of an orally administrable nanoparticle composition containing insulin The orally administrable nanoparticle composition containing insulin was prepared
• the mean particle size was 129.4 nm
• Example 5 Preparation 4 of an orally administrable nanoparticle
• the orally administrable nanoparticle composition containing insulin was prepared
• EudragitTM (Rohm) and Eudragit STM (Rohm) instead of chitosan. According to the
• Example 1 the particle size and degree of dispersion were measured, and as a
• the mean particle size was 194.2 nm, and the degree of dispersion was 0.128.
• the liquid formulation was prepared according to the same method as Example 5
• Example 6 Preparation of an orally administrable nanoparticle containing
• ceftriaxone (1) Preparation of a complex of ceftriaxone and a counter-ion substance
• ceftriaxone formed the complex with the counter-ion substance.
• Example 1 the orally administrable nanoparticle formulation containing ceftriaxone was
• the mean particle size was 199.0 nm
• Example 1 the particle size and degree of dispersion were measured. As a result, the
• mean particle size was 149.4 nm, and the degree of dispersion was 0.124.
• Example 7 Preparation of an orally administrable nanoparticle formulation
• Poloxamer 407TM and the whole mixture was stirred. Then, according to the method of
• Example 1 the orally administrable nanoparticle formulation containing ceftriaxone was
• citric acid was added as an additive to increase the solubility of
• the mean particle size was 290.1 nm
• Lipases are present in vivo, and nanoparticles composed of lipids should be stable against lipases such that insulin entrapped in lipid nanoparticles is not exposed to the exterior and so is not degraded by proteases.
• the formulations of examples and comparative examples were tested for their stability against in vivo enzyme degrading proteins and lipids, pancreatin.
• pancreatin (USP grade). The reaction was performed at 37 ° C for 0, 1, 3 and 5 hours, and the residual amount of insulin was measured and shown in the following Table Table 5: Stability of orally administrable nanoparticle formulations containing insulin against pancreatin
• Examples 1 to 5 formulations containing the insulin-counter-ion complex and the lipid/polymer had high drag entrapping rate and enzyme stability.
• Comparative Example 1 formulation containing only lipid and Comparative Example 4 formulation containing only polymer had extremely low stability against enzyme, that is, the stability of insulin against pancreatin after 5 hours was just less than 20%>.
• Comparative Example 3 formulation prepared by using only microfluidizer in preparing nanoparticle formulation had lower entrapping rate of insulin-counter-ion complex and lower stability against enzyme than Example 1 formulation prepared by first using such solubilizing agent as ethanol and then using microfluidizer.
• Example la nanoparticle formulation prepared by using solubilizing agent without using microfluidizer had high drug entrapping rate and stability against enzyme like Example 1 formulation.
• nanoparticle formulations prepared from insulin not from an
• insulin-counter-ion complex have low drag entrapping rate, and nanoparticle formulations
• nanoparticles of the present invention prepared by forming a
• streptozocin 45 mg/kg of streptozocin (Sigma) was intraperitoneally injected twice to normal male Spraque Dawley rats weighing about 180-220 g at the interval of -two days to produce type I diabetic rats. After one week, the rats were fast for 12 hours, and then, the blood was gathered therefrom and their blood glucose was measured. As a result, the rats having the blood glucose of 300 mg/dl or more were considered type I diabetic rats, and used for the subsequent experiment. The blood glucose was measured using Glucotrand2TM (Roche).
• the nanoparticle liquid formulation of the present invention was confirmed to retain the physiological activity of insulin intact. Therefore, animal experiments were performed using the liquid formulation of the present invention, below.
• Type I diabetic rat was ether-anesthetized, and the abdomen was opened.
• diabetic rat was allowed to move and drink water ad libitum.
• Example la were used.
• the formulation of Comparative Example 1 was the nanoparticle
• Example la had high drag entrapping rate and enzyme stability, but was the
• Example 1 administered at 20 IU/kg via the same route as Example 1, and as reference group 2, the
• insulin injectable solution was subcutaneously injected at 0.2 IU/kg.
• Example la formulation prepared by omitting passing through the
• microfluidizer was about 70%> of that of Example 1.
• the nanoparticle formulation entrapping insulin
• blood glucose reducing activity could be enhanced by
• Example 1 formulation As shown in Fig. 4, administration of Example 1 formulation showed very similar
• the orally administrable nanoparticle liquid formulation containing no insulin was administered via the same route as Example 1, and as the
• insulin injectable solution was subcutaneously injected at 0.2 IU/kg.
• the blood concentration of insulin was less than 20 ⁇ lU/ml in
• Example 1 170 ⁇ lU/ml, and so insulin abso ⁇ tion rate was high, in Example 1 formulation.
• microfluidizer showed about 70 ⁇ lU/ml of insulin abso ⁇ tion rate at the time of maximal
• Example 1 formulation showed increased
• pressure homogenizer are very effective to increase insulin abso ⁇ tion rate in type I diabetic rats.
• Example 1 formulation showed the similar insulin blood concentration profile to subcutaneous injection of 0.2 IU/kg of insulin which is applied to early stage of diabetic patients.
• formulation containing ceftriaxone was administered thereto at 40 mg/kg as ceftriaxone.
• Comparative Example 5 The formulations of Examples 6 and Example 7, and Comparative Example 5 were used for test groups.
• the formulation of Comparative Example 5 is one containing neither ceftriaxone-counter-ion complex nor polymer, and so having lower drag entrapping rate and incapable of controlling drug dissolution compared with that of
• Example 6 Blood was gathered from tail vein at 0, 0.5, 1, 1.5, 2, 3 and 4 hours after the drag administration, and analyzed according to ceftriaxone analysis method of the US Pharmacopoeia. The blood was centrifuged at 3000 ⁇ m for 10 minutes, and pretreated by adding the equal amount of acetonitrile thereto.
• ceftriaxone dissolved in water was administered at 40 mg/kg
• ceftriaxone dissolved in a physiological saline was intravenously injected at 20 mg/kg.
• bioavailability was 22.8% and 35.1% in case of
• ceftriaxone was not nearly absorbed in vivo in
• ceftriaxone was entrapped only in lipid, compared with the groups to which that the
• composition of the present invention has high entrapping rate
• the orally administrable nanoparticle composition of the present invention has high entrapping rate of a water-soluble drag in the nanoparticle, and protects the unstable drug from gastrointestinal enzymes, and has high abso ⁇ tion rate to the gastrointestinal membrane. Therefore, it is extremely useful as drag delivery system for enhancing the bioavailability of water-soluble drags whose oral administration has been limited due to charges thereof.

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