WO2016056728A1 - Improved mesoglycan drug delivery dosage form - Google Patents

Improved mesoglycan drug delivery dosage form Download PDF

Info

Publication number
WO2016056728A1
WO2016056728A1 PCT/KR2015/005638 KR2015005638W WO2016056728A1 WO 2016056728 A1 WO2016056728 A1 WO 2016056728A1 KR 2015005638 W KR2015005638 W KR 2015005638W WO 2016056728 A1 WO2016056728 A1 WO 2016056728A1
Authority
WO
WIPO (PCT)
Prior art keywords
peg
mesoglycan
drug delivery
surfactant
hydrophobic
Prior art date
Application number
PCT/KR2015/005638
Other languages
French (fr)
Korean (ko)
Inventor
정은아
Original Assignee
네오지에이지주식회사
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 네오지에이지주식회사 filed Critical 네오지에이지주식회사
Publication of WO2016056728A1 publication Critical patent/WO2016056728A1/en

Links

Images

Classifications

    • 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
    • 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
    • A61K31/716—Glucans
    • 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
    • A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates

Definitions

  • the present invention relates to an improved mesoglycan drug delivery formulation. More specifically, it relates to mesoglycan drug delivery formulations with increased absorption and bioavailability in the gastrointestinal tract consisting of hydrophobic polymer-surfactant-mesoglycans.
  • Mesoglycans are glycosaminoglycans (GAG) -derived drugs, heparin sulfate (47-57%), dermatan sulfate (25-35%), and chondroitin sulfate (4) derived from aortic / intestinal mucosa derived from animals such as cattle or pigs. ⁇ 8%) and slow moving heparin (2-10%) of the anti-thrombotic effect of the vascular disease treatment.
  • GAG glycosaminoglycans
  • 1 is a diagram showing the chemical structural characteristics of the polymer repeating unit form of the four components of the mesoglycan of the present invention.
  • Mesoglycans have an anticoagulant effect and recently, oral administration of 50-150 mg of mesoglycan is usually used for the prevention and treatment of ischemic brain disease in patients with vascular diseases.
  • mesoglycans have an anticoagulant effect that increases the activity of anti-thrombin III and heparin cofactor II and inhibits the activity of factor VIIa and IIa, and increases the activity of lipoprotein lipase to increase the triglyceride levels in the blood.
  • Lowering the level of low density cholesterol (LDL) and increasing the level of high density cholesterol (HDL) has an anti-hyperlipidemic effect.
  • single dose of mesoglycan shows a fibrin dissolution effect in proportion to the dose, and repeated administration of fibrin shows a continuous dissolution effect but does not affect other hemostatic systems.
  • glycosaminoglycan-based biopolymers such as mesoglycans impair mucosal absorption in the gastrointestinal tract upon oral administration, which suggests that glycosaminoglycans have a molecular weight of greater than 10,000 molecular weight and high negative charge in the polymer. It is due to the nature of the hydrophilic material. On the other hand, such a polymer material is more difficult to absorb the mucous membrane in the gastrointestinal tract under the influence of hydrolysis or enzymatic degradation in the gastrointestinal tract.
  • a delivery agent for a biologically active agent comprising a hydrophilic moiety selected from a dipeptide or tripeptide containing a charged ⁇ -amino acid, a polyamine and a chelating agent, wherein the hydrophobic moiety and the hydrophilic moiety are covalently bonded.
  • heparin of low molecular weight is disclosed as one of the biologically active agents, and deoxycholic acid is disclosed as the hydrophobic site.
  • Charged ⁇ -amino acids also list lysine, arginine, histidine, aspartic acid or glutamic acid.
  • the delivery agent of Formula I was effective in enhancing the absorption of low molecular weight heparin in the body, but there was a problem in the excretion of the delivery agent in the body, so when long-term delivery of a large amount of delivery agent with low molecular weight heparin accumulated in the body There could be side effects due to the safety was a problem.
  • Korean Patent Registration No. 10-1027161, 'Heparin conjugate and preparation method', heparin-DOCA, heparin and N-deoxycholine acid conjugate, heparin-tau, a heparin-taurocholic acid conjugate Rocolinic acid and heparin-litocholine acid, which are conjugates of heparin and litrocholine acids, have been disclosed.
  • These heparin conjugates were intended to solve the problem of accumulating the transfer agent of Formula I in the body by chemically bonding steroid compounds of various choline acids through heparin and amide bonds.
  • the present inventors have developed a mesoglycan drug delivery formulation that solves the above problems, and when the mesoglycan polymer is adsorbed and combined with a hydrophilic surfactant and a hydrophobic surfactant suitable for a hydrophobic polymer such as choline acid,
  • the present invention was completed by confirming through experiments that the gastrointestinal tract absorption of Khan is significantly increased and thus the anti-coagulant activity of the body is continuously increased.
  • the problem to be solved by the present invention is to develop a mesoglycan drug delivery formulation in which the absorption in the gastrointestinal tract is greatly increased and the anti-coagulant activity in the body is also continuously increased.
  • a hydrophilic and hydrophobic surfactant suitable for hydrophobic polymers such as choline acid is used to develop a mesoglycan drug delivery formulation in which a mesoglycan polymer is adsorbed and bound.
  • It is an object of the present invention to provide a mesoglycan drug delivery formulation comprising 1) 100 parts by weight of mesoglycans, 2) 10 to 50 parts by weight of choline acid, and 3) 10 to 100 parts by weight of hydrophilic and hydrophobic surfactants.
  • the hydrophilic surfactant is a nonionic surfactant having an HLB value of 12 or more
  • the hydrophobic surfactant is a nonionic surfactant having an HLB value of 8 or less.
  • the content of the hydrophilic surfactant and the hydrophobic surfactant is characterized in that the weight ratio of 0.8 ⁇ 1.2: 1.2 ⁇ 0.8.
  • the hydrophilic nonionic surfactant is characterized in that the HLB value is 12 or more polyethylene glycol fatty acid ester.
  • the hydrophobic nonionic surfactant is characterized by being a transesterification product of polyethylene glycol having an HLB value of 8 or less.
  • the polyethylene glycol fatty acid ester is PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-12 oleate, PEG-20 oleate, PEG-32 oleate, PEG-15 stearate, PEG -40 stearate, PEG-20 glyceryllaurate, PEG-20 glyceryl oleate, PEG-40 glyceryl laurate, PEG-35 castor oil, PEG-40 hydrogenated castor oil, PEG-30 cholesterol or PEG-25 phyto It is characterized in that at least one selected from sterols.
  • the transesterified product of polyethylene glycol is also selected from PEG-5 hydrogenated castor oil, PEG-7 hydrogenated castor oil, PEG-6 corn oil, PEG-6 almond oil, PEG-6 olive oil or PEG-6 palm kernel oil. It is characterized by one or more.
  • the effect of the present invention is to provide a mesoglycan drug delivery formulation in which the absorption in the gastrointestinal tract is greatly increased and the anti-coagulant activity in the body is also continuously increased.
  • a hydrophilic surfactant suitable for hydrophobic polymer materials such as choline acid and a hydrophobic surfactant are used to provide a mesoglycan drug delivery formulation in which the mesoglycan polymer is adsorbed and bound.
  • FIG. 1 is a diagram showing the polymer structure of heparin sulfate, dermatan sulfate, chondroitin sulfate and heparin constituting mesoglycans.
  • the chemical structural features of the polymer repeating unit form of the four components of the mesoglycan of the present invention are shown.
  • the present invention relates to a mesoglycan drug delivery formulation comprising 1) 100 parts by weight of mesoglycans, 2) 10 to 50 parts by weight of choline acid, and 3) 10 to 100 parts by weight of hydrophilic and hydrophobic surfactants.
  • the active agent relates to a mesoglycan drug delivery formulation characterized in that the nonionic surfactant has an HLB value of 12 or more, and the hydrophobic surfactant is a nonionic surfactant having an HLB value of 8 or less.
  • Mesoglycans are glycosaminoglycans (GAG) -derived drugs, heparin sulfate (47-57%), dermatan sulfate (25-35%), and chondroitin sulfate (4) derived from aortic / intestinal mucosa derived from animals such as cattle or pigs. ⁇ 8%) and slow moving heparin (2-10%) of the anti-thrombotic effect of the vascular disease treatment.
  • GAG glycosaminoglycans
  • the mesoglycan has a carboxyl group (COO ⁇ ) and a sulfonic acid group (SO 3 ⁇ ) as shown in FIG. 1, and has a lot of negatively charged electric charges, indicating hydrophilicity.
  • dermatan sulfate is similar to that of chondroitin sulfate (CS), which contains a disaccharide of [IdoA ⁇ (1 ⁇ 3) GalNAc] as a repeating unit, and chondroitin sulfate (CS) is [GlcA ⁇ (1 ⁇ 3) GalNAc]. Include disaccharide in repeat units. In addition, these disaccharide repeat units are bound by ⁇ (1 ⁇ 4) linkages.
  • Heparin is similar to heparin sulfate (HS) structure, but compared to heparin sulfate, heparin sulfate is 70% higher in heparin sulfate than 40% heparin sulfate, and GlcNSO 3 H is less than 50% heparin sulfate within 80% heparin sulfate. (1.8 ⁇ 2.6% -Heparin: 0.8 ⁇ 1.6% -Heparan sulfate) is higher than heparin sulfate.
  • Mesoglycans with this composition have a wide range of biological functions such as organ formation, growth control, cytokine activity regulation, tissue binding, signaling, inflammation and interaction with pathogens, cancer cell growth and invasion, etc. It has been used as a material.
  • mesoglycan is used as a drug for treating symptomatic improvement of patients with chronic senile cerebrovascular disease accompanied with a thrombotic risk or cerebral insufficiency due to cerebral atherosclerosis.
  • the antithrombotic pharmacological mechanism is to activate plasminogen by increasing plasminogen utilization by activating plasminogen with plasmin, a fibrinolytic enzyme.
  • the content of choline acid relative to 100 parts by weight of mesoglycan is 10 to 50 parts by weight, and preferably 15 to 30 parts by weight.
  • the content of the hydrophilic and hydrophobic surfactant is 10 to 100 parts by weight, preferably 20 to 50 parts by weight.
  • Mesoglycans and choline acids are adsorbed and bound to hydrophilic and hydrophobic surfactants, and their shape may thrive in accordance with the pH of the solution in the body.
  • the content of the hydrophilic surfactant and hydrophobic surfactant used in the present invention is 0.8 to 1.2: 1.2 to 0.8 in molar ratio, more preferably 0.9 to 1.1: 1.1 to 0.9 and most preferably 1: 1. It is therefore most preferred when the hydrophilic and hydrophobic surfactants are at the same molar ratio.
  • Hydrophilic mesoglycans are adsorbed to hydrophilic surfactants and hydrophobic choline acid is adsorbed to hydrophobic surfactants.
  • hydrophilic and hydrophobic surfactants form micelles in the body and are distributed in the body fluid.
  • mesoglycan which was difficult to absorb gastrointestinal mucosa due to hydrophilic substances, the affinity between hydrophobic choline acid and gastrointestinal mucosa was increased due to the effect of hydrophobic choline acid and surfactant adsorbed on mesoglycan when switching to the mesoglycan drug formulation of the present invention.
  • absorbed mesoglycan drug delivery formulations mesoglycans and choline acids in the blood are adsorbed with hydrophilic and hydrophobic surfactants and are readily dissolved in the blood under the influence of surfactants.
  • mesoglycans and choline acids are adsorbed and dispersed in the blood with surfactants, the mesoglycans can be continuously released under the influence of the adsorbed surfactant, and both mesoglycans and choline acids can be easily excreted through the kidneys. Can be.
  • the mesoglycan drug delivery formulation of the present invention can solve the side effects of the conventionally developed delivery agent of Formula I in the body and also solve the safety problem as a drug drug delivery agent such as the case of heparin conjugates. That is a safe formulation.
  • the hydrophilic nonionic surfactant is characterized by being a polyethylene glycol fatty acid ester having an HLB value of 12 or more, and the hydrophobic nonionic surfactant is characterized by being a transesterification product of polyethylene glycol having an HLB value of 8 or less.
  • the polyethylene glycol fatty acid ester is PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-12 oleate, PEG-20 oleate, PEG-32 oleate, PEG-15 stearate, PEG -40 stearate, PEG-20 glyceryllaurate, PEG-20 glyceryl oleate, PEG-40 glyceryl laurate, PEG-35 castor oil, PEG-40 hydrogenated castor oil, PEG-30 cholesterol or PEG-25 phyto One or more selected from sterols may be used.
  • the transesterified product of polyethylene glycol is also selected from PEG-5 hydrogenated castor oil, PEG-7 hydrogenated castor oil, PEG-6 corn oil, PEG-6 almond oil, PEG-6 olive oil or PEG-6 palm kernel oil. 1 or more types can be used.
  • Mesoglycan drug delivery formulations of the Khan / surfactant / deoxycholine acid complexes were prepared.
  • Mesoglycan drug delivery formulations of the Khan / surfactant / deoxycholine acid complexes were prepared.
  • a mesoglycan drug delivery formulation disclosed in Korean Patent Registration No. 10-679677 was prepared by mixing 300 mg of N ⁇ -deoxycholyl-L-lysine-methylester with 1000 mg of mesoglycans.
  • Rats purified for one week in an animal feeding room were anesthetized with zoletil (animal anesthetic), collected from the vena cava, and mixed with an anticoagulant, 0.109 M trisodium citrate, to a ratio of 1: 9 (v / v).
  • Blood was used for the experiment by separating the platelet-free plasma (PPP).
  • the anti-factor VIIa activity was measured in the drug delivery formulations prepared in the preparation examples and the comparative examples in which isolated plasma (PPP) was dissolved in purified water at a constant concentration (final concentration 0, 4, 8, 12, 16 ⁇ g / ml). Were mixed at a ratio of 4: 1, the standard curve was prepared using a coagulation analyzer, and anti-factor VIIa activity (U / mg) was measured.
  • the anti-factor IIa activity was measured by mixing the drug delivery formulations prepared in the preparation examples and the comparative examples, which were dissolved in separated plasma (PPP) and purified water at constant concentrations (final concentrations 0, 4, 8, 12, 16 ⁇ g / ml). Then, add 800 ⁇ l of pH 8.4 buffer and 100 ⁇ l of anti thrombin (1IU / ml) to make a sample solution, and heat 400 ⁇ l of this at 37 ° C for 4 minutes, add 200 ⁇ l of factor IIa (5U / ml), and mix well at 37 ° C. After precisely warming for 30 seconds, add 400 ⁇ l of substrate solution S-2238 (1.25 mmol / L), which was warmed at 37 ° C., and mix well.
  • PPP separated plasma
  • Table 1 shows the results of measuring the effects of in vitro anti-factor IIa and anti-factor VIIa of the drug delivery formulation. It was confirmed that the drug delivery formulations prepared in Preparation Example 1 and Preparation Example 2 showed a relatively good effect. In addition, the activity of anti factor Xa per mg of polymer was calculated as U (unit) using the standard curve.
  • Mesoglycan drug delivery formulations in which the hydrophilic and hydrophobic surfactants were added to and mixed with mesoglycans in the in vitro test of Example 1 were compared to those of the mesoglycan drug delivery formulations prepared in Preparation Examples 1 and 2 described above. It showed excellent anti-factor IIa and anti-factor VIIa effects.
  • hydrophobic compounds such as choline acid, deoxycholic acid (DOCA) and taurocholine acid were used to determine what kind of hydrophobic polymer mixed and adsorbed to mesoglycan and surfactant showed the best anti-coagulation effect.
  • Mesoglycan drug delivery formulations were prepared using polymers and their activity was measured in vitro and in vivo.
  • Mesoglycan drug delivery formulations of the Khan / surfactant / deoxycholine acid complexes were prepared.
  • Mesoglycan drug delivery formulations of the / taurocholine acid complex were prepared.
  • the mesoglycan drug delivery formulations prepared in Examples 3 and 4 and Comparative Examples 3 and 4 were dissolved or suspended in water and orally administered to rats purified for one week in an animal rearing room. Blood samples were collected for 15 minutes, 30 minutes, 1 hour, and 5 hours after oral administration in the abdominal and orbital veins, and the anticoagulant 0.109 M trisodium citrate was mixed with blood at a ratio of 1: 9 (v / v). Blood was used for the experiment by separating the platelet-free plasma (PPP). Hemosil liquid heparin kit was used for anti-factor VIIa activity against plasma (PPP) from which platelets obtained by centrifugation were removed.
  • PPP platelet-free plasma
  • Anti-factor IIa activity was measured by adding 800 ⁇ l of pH 8.4 buffer and 100 ⁇ l of antithrombin (1IU / ml) to 100 ⁇ l of platelet-free plasma (PPP) obtained by centrifugation. Heparin standard solution and test method except the test solution were performed in the same manner as the in vitro experimental anti-factor IIa activity measurement.
  • Table 2 shows the anti-factor VIIa in plasma after 15 minutes, 30 minutes, 1 hour, and 5 hours by oral administration of the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 and Comparative Examples 3 and 4 to rats. Activity was measured. As a result, the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 were highest at 30 minutes, whereas the mesoglycan drug delivery formulations prepared in Preparation Example 3 were observed at 60 minutes, but the preparation comparison Mesoglycan drug delivery formulations prepared in 4 showed little effect in vivo.
  • the mesoglycan drug delivery formulation prepared in Preparation Example 3 showed the highest anti-factor VIIa activity of 0.72 U / ml at 30 minutes, and the mesoglycan drug delivery formulation prepared in Preparation Example 4 was used at 30 minutes. The highest anti-factor VIIa activity of 0.68 U / ml was observed.
  • Table 3 shows the anti-factor IIa in plasma after 15 minutes, 30 minutes, 1 hour, and 5 hours by oral administration of the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 and Comparative Examples 3 and 4. Activity was measured. As a result, the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 were highest at 30 minutes, whereas the mesoglycan drug delivery formulations prepared in Preparation Example 3 were observed at 60 minutes, but the preparation comparison Mesoglycan drug delivery formulations prepared in 4 showed little effect in vivo.
  • the mesoglycan drug delivery formulation prepared in Preparation Example 3 showed the highest anti-factor IIa activity at 0.34 U / ml at 30 minutes, and the mesoglycan drug delivery formulation prepared at Preparation Example 4 was used at 30 minutes. The highest anti-factor IIa activity, 0.35 U / ml, was observed.
  • the mesoglycan drug delivery formulations prepared in Examples 3 and 4 and Comparative Examples 3 and 4 were dissolved or suspended in water and orally administered to rats purified for one week in an animal rearing room. Next, blood was collected for 15 minutes, 30 minutes, 1 hour, and 5 hours after oral administration from the abdominal and orbital veins, and the anticoagulant 0.109M trisodium citrate and blood were mixed at a ratio of 1: 9 (v / v). Blood was used for the experiment by separating the platelet-free plasma (PPP). APTT for platelet-depleted plasma (PPP) obtained by centrifugation was measured by an automated blood coagulation analyzer (ACL-7000) using an ACL-assay reagent kit.
  • PPP platelet-free plasma
  • ACL-7000 automated blood coagulation analyzer
  • Table 4 shows the results of measuring APTT in plasma after 15 minutes, 30 minutes, 1 hour and 5 hours by oral administration of the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 and Comparative Examples 3 and 4. .
  • APTT was extended from 30 minutes to 46.3 seconds and from 1 hour to 57.5 seconds.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Inorganic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)

Abstract

The present invention relates to a mesoglycan drug delivery dosage form in which 1) 100 parts by weight of mesoglycan, 2) 10-50 parts by weight of cholic acid and 3) 10-100 parts by weight of hydrophilic and hydrophobic surfactants are adsorbed and coupled, wherein the hydrophilic surfactant is a nonionic surfactant having an HLB value of 12 or more and the hydrophobic surfactant is a nonionic surfactant having an HLB value of 8 or less.

Description

향상된 메소글리칸 약물 전달 제형Enhanced Mesoglycan Drug Delivery Formulations
본 발명은 향상된 메소글리칸 약물 전달 제형에 관한 것이다. 더욱 상세하게는 소수성 고분자-계면활성제-메소글리칸으로 구성된 위장관 내 흡수성 및 생체 이용률이 증가된 메소글리칸 약물 전달 제형에 관한 것이다.The present invention relates to an improved mesoglycan drug delivery formulation. More specifically, it relates to mesoglycan drug delivery formulations with increased absorption and bioavailability in the gastrointestinal tract consisting of hydrophobic polymer-surfactant-mesoglycans.
메소글리칸은 글리코스아미노글리칸(GAG) 유래 의약품으로 소 또는 돼지 등의 동물 유래 대동맥/장점막 등에서 추출한 헤파린 설페이트(47~57%), 더마탄 설페이트(25~35%), 콘드로이틴 설페이트(4~8%) 및 slow moving 헤파린(2~10%)의 혼합 성분으로 구성된 항-혈전 효과가 있는 혈관 질환 치료제이다.Mesoglycans are glycosaminoglycans (GAG) -derived drugs, heparin sulfate (47-57%), dermatan sulfate (25-35%), and chondroitin sulfate (4) derived from aortic / intestinal mucosa derived from animals such as cattle or pigs. ~ 8%) and slow moving heparin (2-10%) of the anti-thrombotic effect of the vascular disease treatment.
도 1은 본 발명의 메소글리칸의 4가지 성분의 고분자 반복단위 형태의 화학적 구조 특징을 나타낸 도면이다. 1 is a diagram showing the chemical structural characteristics of the polymer repeating unit form of the four components of the mesoglycan of the present invention.
메소글리칸은 항응고 효과를 나타내고 있으며 최근 혈관질환 환자의 허혈성 뇌질환 예방 및 치료에 통상 50∼150mg의 메소글리칸을 경구 투여하여 사용하고 있다. Mesoglycans have an anticoagulant effect and recently, oral administration of 50-150 mg of mesoglycan is usually used for the prevention and treatment of ischemic brain disease in patients with vascular diseases.
이러한 메소글리칸은 항-트롬빈 Ⅲ와 헤파린 보조인자Ⅱ의 활성을 증가시켜 factor Ⅹa와 Ⅱa의 활성을 저해하는 항-응고 효과를 나타내며, 지단백 리파아제(lipoprotein lipase)의 활성을 증가시켜 혈중 중성지방과 저밀도 콜레스테롤(LDL)의 수치를 낮추고 고밀도 콜레스테롤(HDL) 수치를 증가시켜 항-고지혈증 효과를 나타낸다.These mesoglycans have an anticoagulant effect that increases the activity of anti-thrombin III and heparin cofactor II and inhibits the activity of factor VIIa and IIa, and increases the activity of lipoprotein lipase to increase the triglyceride levels in the blood. Lowering the level of low density cholesterol (LDL) and increasing the level of high density cholesterol (HDL) has an anti-hyperlipidemic effect.
또한 메소글리칸 1회 투여시에도 용량에 비례하여 피브린 용해 효과를 나타내며 반복 투여시 피브린의 지속적인 용해 효과를 나타내나 다른 지혈체계에 영향을 주지 않는다. In addition, single dose of mesoglycan shows a fibrin dissolution effect in proportion to the dose, and repeated administration of fibrin shows a continuous dissolution effect but does not affect other hemostatic systems.
그러나 메소글리칸과 같은 글리코스아미노글리칸 계열의 생체 고분자 물질들은 경구 투여시 위장관 내의 점막 흡수에 장애가 있으며 이는 글리코스아미노글리칸 계열의 물질이 분자량 10,000 이상의 거대 분자 크기와 고분자 내의 높은 음전하를 띤 친수성 물질의 특성에 기인한다. 한편으로는 이러한 고분자 물질은 위장관 내의 가수분해 또는 효소분해 등의 영향으로 위장관 내의 점막 흡수를 더욱 어렵게 하는 것이다. However, glycosaminoglycan-based biopolymers such as mesoglycans impair mucosal absorption in the gastrointestinal tract upon oral administration, which suggests that glycosaminoglycans have a molecular weight of greater than 10,000 molecular weight and high negative charge in the polymer. It is due to the nature of the hydrophilic material. On the other hand, such a polymer material is more difficult to absorb the mucous membrane in the gastrointestinal tract under the influence of hydrolysis or enzymatic degradation in the gastrointestinal tract.
이러한 메소글리칸과 같은 헤파린 고분자의 낮은 경구 위장관 흡수율을 극복하기 위하여 여러가지 제형 등이 개시되어 있다. Various formulations have been disclosed to overcome low oral gastrointestinal absorption of heparin polymers such as mesoglycans.
대한민국 등록특허공보 제10-679677호 '생물학적 활성제의 점막흡수 증강을 위한 전달제'에서는 '(1) 담즙산, 스테롤, 담즙산 유도체 및 스테롤 유도체에서 선택되는 소수성 부위, (2) 전하를 띤 α-아미노산, 상기 전하를 띤 α-아미노산을 함유하는 다이 펩타이드 또는 트리 펩타이드, 폴리아민 및 킬레이트화제에서 선택된 친수성 부위로 이루어진 생물학적 활성제의 전달제에 있어서, 상기 소수성 부위와 상기 친수성 부위가 공유결합하고 있는 것을 특징으로 하는 생물학적 활성제의 전달제'를 개시하고 있다. Republic of Korea Patent Publication No. 10-679677 'Delivery agent for enhancing the mucosal absorption of biologically active agent' in (1) hydrophobic sites selected from bile acids, sterols, bile acid derivatives and sterol derivatives, (2) charged α-amino acid A delivery agent for a biologically active agent comprising a hydrophilic moiety selected from a dipeptide or tripeptide containing a charged α-amino acid, a polyamine and a chelating agent, wherein the hydrophobic moiety and the hydrophilic moiety are covalently bonded. To a delivery agent of a biologically active agent.
이때 생물학적 활성제의 하나로 저분자량의 헤파린을 개시하고 있으며 상기 소수성 부위로는 데옥시콜린산 등을 개시하고 있다. 또한 전하를 띤 α-아미노산은 라이신, 아르기닌, 히스티딘, 아스파트산 또는 글루타민산을 열거하고 있다. At this time, heparin of low molecular weight is disclosed as one of the biologically active agents, and deoxycholic acid is disclosed as the hydrophobic site. Charged α-amino acids also list lysine, arginine, histidine, aspartic acid or glutamic acid.
이러한 소수성 부위와 친수성 부위가 서로 공유결합을 통해 결합됨으로써 하기식 Ⅰ과 같은 Nα-데옥시콜릴-L-라이신-메틸에스테르를 생성시키고 이를 저분자량 헤파린과 함께 경구 투여함으로써 동물모델 내의 항-factor Ⅹa 활성의 증가를 개시하고 있다.These hydrophobic and hydrophilic sites are covalently bonded to each other to produce N α -deoxycholyl-L-lysine-methyl ester as shown in the following formula I, which is orally administered with low molecular weight heparin to prevent the anti-factor in the animal model. Increasing VIIa activity is initiated.
Figure PCTKR2015005638-appb-I000001
식 Ⅰ
Figure PCTKR2015005638-appb-I000001
Equation Ⅰ
그러나 상기 식 Ⅰ의 전달제는 저분자량 헤파린의 체내 흡수 증진에는 효과적이었으나 상기 전달제의 체내 배설에 문제가 있어 장기간 다량의 전달제를 저분자량의 헤파린과 함께 경구 투여하는 경우에는 전달제의 체내 축적에 따른 부작용이 발생할 수 있어 그 안전성에 문제가 있었던 것이다. However, the delivery agent of Formula I was effective in enhancing the absorption of low molecular weight heparin in the body, but there was a problem in the excretion of the delivery agent in the body, so when long-term delivery of a large amount of delivery agent with low molecular weight heparin accumulated in the body There could be side effects due to the safety was a problem.
이와 같은 체내 배설 문제를 해결하기 위해 대한민국 특허등록 제10-1027161호 '헤파린 결합체 및 제조방법'에서는 헤파린과 N-데옥시콜린산 결합체인 헤파린-DOCA, 헤파린과 타우로콜린산 결합체인 헤파린-타우로콜린산 및 헤파린과 리토콜린산 결합체인 헤파린-리토콜린산 등을 개시한 바 있다. 이들 헤파린 결합체는 각종 콜린산류의 스테로이드 화합물을 헤파린과 아마이드 결합을 통해 화학 결합시킴으로써 체내 상기 식 Ⅰ의 전달제가 축적되는 문제를 해결하고자 한 것이었다. In order to solve the problem of excretion of the body, Korean Patent Registration No. 10-1027161, 'Heparin conjugate and preparation method', heparin-DOCA, heparin and N-deoxycholine acid conjugate, heparin-tau, a heparin-taurocholic acid conjugate Rocolinic acid and heparin-litocholine acid, which are conjugates of heparin and lithocholine acids, have been disclosed. These heparin conjugates were intended to solve the problem of accumulating the transfer agent of Formula I in the body by chemically bonding steroid compounds of various choline acids through heparin and amide bonds.
또한 상기 특허문헌의 실시예에는 헤파린-DOCA 결합체의 항-응고 능력이 가장 우수함을 개시하고 있다.In addition, the examples of the patent document discloses the best anti-coagulation ability of the heparin-DOCA conjugate.
그러나 상기 특허문헌에 개시된 헤파린-DOCA 결합체의 경우 그 화학적 합성을 위해서는 촉매 반응물질로서 디사이클로헥실카르보디이미드(DCC)와 하이드록시숙신이미드를 DMF 용매에 녹여 사용하여야 하는 바, 이러한 촉매 반응물질과 DMF 등의 용매를 완전히 제거하지 않고서는 안전성의 문제가 있어 헤파린과 같은 의약품 약물 전달제로 사용하기 힘든 문제가 있었던 것이다. However, in the case of the heparin-DOCA conjugate disclosed in the patent document, for the chemical synthesis, dicyclohexylcarbodiimide (DCC) and hydroxysuccinimide must be dissolved in a DMF solvent and used as a catalyst reactant. Without completely removing solvents such as DMF and DMF, there was a problem of safety, which is difficult to use as a drug delivery agent such as heparin.
따라서 본 발명자들은 상기와 같은 문제점 등을 해결한 메소글리칸 약물전달 제형을 개발하던 중 콜린산과 같은 소수성 고분자 물질에 적합한 친수성 계면활성제와 소수성 계면활성제를 사용하여 메소글리칸 고분자를 흡착 결합시키면 메소글리칸의 위장관 흡수가 현격히 증가되고 이에 따른 체내의 항-응고 활성 역시 지속적으로 증가됨을 실험을 통해 확인함으로써 본 발명을 완성하게 된 것이다.Therefore, the present inventors have developed a mesoglycan drug delivery formulation that solves the above problems, and when the mesoglycan polymer is adsorbed and combined with a hydrophilic surfactant and a hydrophobic surfactant suitable for a hydrophobic polymer such as choline acid, The present invention was completed by confirming through experiments that the gastrointestinal tract absorption of Khan is significantly increased and thus the anti-coagulant activity of the body is continuously increased.
따라서 본 발명이 해결하고자 하는 과제는 위장관 내 흡수가 현격히 증가되고 체내 항-응고 활성 역시 지속적으로 증가되는 메소글리칸 약물 전달 제형을 개발코자 한 것이다. 콜린산과 같은 소수성 고분자 물질에 적합한 친수성 계면활성제와 소수성 계면활성제를 사용하여 메소글리칸 고분자를 흡착 결합시킨 메소글리칸 약물전달 제형을 개발코자 한 것이다.Accordingly, the problem to be solved by the present invention is to develop a mesoglycan drug delivery formulation in which the absorption in the gastrointestinal tract is greatly increased and the anti-coagulant activity in the body is also continuously increased. A hydrophilic and hydrophobic surfactant suitable for hydrophobic polymers such as choline acid is used to develop a mesoglycan drug delivery formulation in which a mesoglycan polymer is adsorbed and bound.
본 발명의 목적은 1) 100 중량부의 메소글리칸, 2) 10~50 중량부의 콜린산 및 3) 10~100 중량부의 친수성 및 소수성 계면활성제를 흡착 결합시킨 메소글리칸 약물 전달 제형에 있어서, 상기 친수성 계면활성제는 HLB 밸류가 12 이상인 비이온성 계면활성제이고, 상기 소수성 계면활성제는 HLB 밸류가 8 이하인 비이온성 계면활성제임을 특징으로 하는 메소글리칸 약물전달 제형을 제공하는 것이다. It is an object of the present invention to provide a mesoglycan drug delivery formulation comprising 1) 100 parts by weight of mesoglycans, 2) 10 to 50 parts by weight of choline acid, and 3) 10 to 100 parts by weight of hydrophilic and hydrophobic surfactants. The hydrophilic surfactant is a nonionic surfactant having an HLB value of 12 or more, and the hydrophobic surfactant is a nonionic surfactant having an HLB value of 8 or less.
이때 상기 친수성 계면활성제와 상기 소수성 계면활성제의 함량은 중량비로 0.8~1.2 : 1.2~0.8 임을 특징으로 한다. At this time, the content of the hydrophilic surfactant and the hydrophobic surfactant is characterized in that the weight ratio of 0.8 ~ 1.2: 1.2 ~ 0.8.
상기 친수성 비이온성 계면활성제는 HLB 밸류가 12 이상인 폴리에틸렌글리콜지방산에스테르임을 특징으로 한다. 상기 소수성 비이온성 계면활성제는 HLB 밸류가 8 이하인 폴리에틸렌글리콜의 트랜스에스테르화물(transesterification product)임을 특징으로 한다. The hydrophilic nonionic surfactant is characterized in that the HLB value is 12 or more polyethylene glycol fatty acid ester. The hydrophobic nonionic surfactant is characterized by being a transesterification product of polyethylene glycol having an HLB value of 8 or less.
또한 상기 폴리에틸렌글리콜지방산에스테르는 PEG-12 라우레이트, PEG-20 라우레이트, PEG-32 라우레이트, PEG-12 올레이트, PEG-20 올레이트, PEG-32 올레이트, PEG-15 스테아레이트, PEG-40 스테아레이트, PEG-20 글리세릴라우레이트, PEG-20 글리세릴올레이트, PEG-40 글리세릴라우레이트, PEG-35 캐스터오일, PEG-40 수소화 캐스터오일, PEG-30 콜레스테롤 또는 PEG-25 파이토스테롤에서 선택된 1종 이상임을 특징으로 한다. In addition, the polyethylene glycol fatty acid ester is PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-12 oleate, PEG-20 oleate, PEG-32 oleate, PEG-15 stearate, PEG -40 stearate, PEG-20 glyceryllaurate, PEG-20 glyceryl oleate, PEG-40 glyceryl laurate, PEG-35 castor oil, PEG-40 hydrogenated castor oil, PEG-30 cholesterol or PEG-25 phyto It is characterized in that at least one selected from sterols.
또한 상기 폴리에틸렌글리콜의 트랜스에스테르화물은 PEG-5 수소화 캐스터오일, PEG-7 수소화 캐스터오일, PEG-6 콘 오일, PEG-6 아몬드 오일, PEG-6 올리브 오일 또는 PEG-6 팜 케르넬 오일에서 선택된 1종 이상임을 특징으로 한다.The transesterified product of polyethylene glycol is also selected from PEG-5 hydrogenated castor oil, PEG-7 hydrogenated castor oil, PEG-6 corn oil, PEG-6 almond oil, PEG-6 olive oil or PEG-6 palm kernel oil. It is characterized by one or more.
본 발명의 효과는 위장관 내 흡수가 현격히 증가되고 체내 항-응고 활성 역시 지속적으로 증가되는 메소글리칸 약물 전달 제형을 제공하는 것이다. 콜린산과 같은 소수성 고분자 물질에 적합한 친수성 계면활성제와 소수성 계면활성제를 사용하여 메소글리칸 고분자를 흡착 결합시킨 메소글리칸 약물전달 제형을 제공하는 것이다.The effect of the present invention is to provide a mesoglycan drug delivery formulation in which the absorption in the gastrointestinal tract is greatly increased and the anti-coagulant activity in the body is also continuously increased. A hydrophilic surfactant suitable for hydrophobic polymer materials such as choline acid and a hydrophobic surfactant are used to provide a mesoglycan drug delivery formulation in which the mesoglycan polymer is adsorbed and bound.
도 1은 메소글리칸을 구성하는 헤파린설페이트, 더마탄설페이트, 콘드로이틴설페이트 및 헤파린의 고분자 구조를 나타낸 도면이다. 상기 도면 내에는 본 발명의 메소글리칸의 4가지 성분의 고분자 반복단위 형태의 화학적 구조 특징을 나타내고 있다.1 is a diagram showing the polymer structure of heparin sulfate, dermatan sulfate, chondroitin sulfate and heparin constituting mesoglycans. In the figure, the chemical structural features of the polymer repeating unit form of the four components of the mesoglycan of the present invention are shown.
본 발명은 1) 100 중량부의 메소글리칸, 2) 10~50 중량부의 콜린산 및 3) 10~100 중량부의 친수성 및 소수성 계면활성제를 흡착 결합시킨 메소글리칸 약물 전달 제형에 있어서, 상기 친수성 계면활성제는 HLB 밸류가 12 이상인 비이온성 계면활성제이고, 상기 소수성 계면활성제는 HLB 밸류가 8 이하인 비이온성 계면활성제임을 특징으로 하는 메소글리칸 약물전달 제형에 관한 것이다. The present invention relates to a mesoglycan drug delivery formulation comprising 1) 100 parts by weight of mesoglycans, 2) 10 to 50 parts by weight of choline acid, and 3) 10 to 100 parts by weight of hydrophilic and hydrophobic surfactants. The active agent relates to a mesoglycan drug delivery formulation characterized in that the nonionic surfactant has an HLB value of 12 or more, and the hydrophobic surfactant is a nonionic surfactant having an HLB value of 8 or less.
이하 본 발명의 전달 약물인 메소글리칸에 대해 더욱 상세히 설명한다. Hereinafter, the delivery drug mesoglycan of the present invention will be described in more detail.
메소글리칸은 글리코스아미노글리칸(GAG) 유래 의약품으로 소 또는 돼지 등의 동물 유래 대동맥/장점막 등에서 추출한 헤파린 설페이트(47~57%), 더마탄 설페이트(25~35%), 콘드로이틴 설페이트(4~8%) 및 slow moving 헤파린(2~10%)의 혼합 성분으로 구성된 항-혈전 효과가 있는 혈관 질환 치료제이다.Mesoglycans are glycosaminoglycans (GAG) -derived drugs, heparin sulfate (47-57%), dermatan sulfate (25-35%), and chondroitin sulfate (4) derived from aortic / intestinal mucosa derived from animals such as cattle or pigs. ~ 8%) and slow moving heparin (2-10%) of the anti-thrombotic effect of the vascular disease treatment.
또한 메소글리칸 내에는 도 1에 나타난 바와 같이 카르복실기(COO-)와 술폰산기(SO3 -)를 지니고 있고 전기적으로 많은 음전하를 띠고 있어 친수성 성질을 나타낸다. In addition, the mesoglycan has a carboxyl group (COO − ) and a sulfonic acid group (SO 3 − ) as shown in FIG. 1, and has a lot of negatively charged electric charges, indicating hydrophilicity.
더마탄 설페이트(DS)의 구조는 콘드로이틴 설페이트(CS)와 비슷하며 [IdoAα(1→3)GalNAc]의 이당을 반복 단위로 포함하고 있으며 콘드로이틴 설페이트(CS)는 [GlcAβ(1→3)GalNAc] 이당을 반복 단위로 포함한다. 또한 이들 이당 반복단위는 β(1→4) 연결고리로 결합되어있다.The structure of dermatan sulfate (DS) is similar to that of chondroitin sulfate (CS), which contains a disaccharide of [IdoAα (1 → 3) GalNAc] as a repeating unit, and chondroitin sulfate (CS) is [GlcAβ (1 → 3) GalNAc]. Include disaccharide in repeat units. In addition, these disaccharide repeat units are bound by β (1 → 4) linkages.
헤파린은 헤파린 설페이트(HS) 구조와 비슷하나 헤파린 설페이트에 비해 이당 구성중 Iduronic acid의 비중이 70%로 40%인 헤파린 설페이트에 비해 높고 GlcNSO3H의 경우 헤파린 80% 이내에 비해 헤파린 설페이트 50% 이내로 sulfation (1.8~2.6%-Heparin:0.8~1.6%-Heparan sulfate)의 비중이 헤파린 설페이트보다 높다.Heparin is similar to heparin sulfate (HS) structure, but compared to heparin sulfate, heparin sulfate is 70% higher in heparin sulfate than 40% heparin sulfate, and GlcNSO 3 H is less than 50% heparin sulfate within 80% heparin sulfate. (1.8 ~ 2.6% -Heparin: 0.8 ~ 1.6% -Heparan sulfate) is higher than heparin sulfate.
이와 같은 조성을 지닌 메소글리칸은 장기 형성, 성장 제어, 사이토카인 활성 조절, 조직의 결합, 신호, 염증 및 병원균과의 상호 작용, 암세포의 성장과 침윤 등과 같은 다양한 생물학적 기능 때문에 의약품, 화장품 및 기능성 식품 소재 등으로 활용되어왔다. Mesoglycans with this composition have a wide range of biological functions such as organ formation, growth control, cytokine activity regulation, tissue binding, signaling, inflammation and interaction with pathogens, cancer cell growth and invasion, etc. It has been used as a material.
또한 메소글리칸은 혈전 위험성이 수반되는 만성 노인성 뇌혈관 질환 환자 또는 뇌동맥경화로 인한 뇌기능 부전 환자의 증상개선 치료 약물로 사용되고 있다. 항혈전 약리 기전은 플라스미노겐을 섬유소 용해효소인 플라스민으로 활성화시켜 플라스미노겐 이용률을 증가시켜 섬유소 용해 과정을 활성화시키는 것이다.In addition, mesoglycan is used as a drug for treating symptomatic improvement of patients with chronic senile cerebrovascular disease accompanied with a thrombotic risk or cerebral insufficiency due to cerebral atherosclerosis. The antithrombotic pharmacological mechanism is to activate plasminogen by increasing plasminogen utilization by activating plasminogen with plasmin, a fibrinolytic enzyme.
이하 본 발명을 더욱 상세히 설명한다. Hereinafter, the present invention will be described in more detail.
본 발명에서 메소글리칸 100 중량부에 대한 콜린산의 함량은 10~50 중량부이며 바람직하게는 15~30 중량부이다. 또한 친수성 및 소수성 계면활성제의 함량은 10~100 중량부이며 바람직하게는 20~50 중량부이다. In the present invention, the content of choline acid relative to 100 parts by weight of mesoglycan is 10 to 50 parts by weight, and preferably 15 to 30 parts by weight. In addition, the content of the hydrophilic and hydrophobic surfactant is 10 to 100 parts by weight, preferably 20 to 50 parts by weight.
메소글리칸과 콜린산은 친수성 및 소수성 계면활성제에 흡착 결합되어 있으며 체내에서 용액의 pH 등에 따라 그 형상이 번화할 수 있으며 필요에 따라 마이셀을 형성하여 체액 내에서 쉽게 분산 용해되는 것이다. Mesoglycans and choline acids are adsorbed and bound to hydrophilic and hydrophobic surfactants, and their shape may thrive in accordance with the pH of the solution in the body.
본 발명에 사용되는 친수성 계면활성제와 소수성 계면활성제의 함량은 몰 비로 0.8~1.2 : 1.2~0.8이며 더욱 바람직하게는 0.9~1.1 : 1.1~0.9이고 최고로 바람직하게는 1:1이다. 따라서 친수성 계면활성제와 소수성 계면활성제가 동일한 몰 비일 때가 가장 바람직하다. The content of the hydrophilic surfactant and hydrophobic surfactant used in the present invention is 0.8 to 1.2: 1.2 to 0.8 in molar ratio, more preferably 0.9 to 1.1: 1.1 to 0.9 and most preferably 1: 1. It is therefore most preferred when the hydrophilic and hydrophobic surfactants are at the same molar ratio.
친수성 계면활성제에는 친수성 메소글리칸이 흡착되며 소수성 계면활성제에는 소수성 콜린산이 흡착된다. 또한 친수성 계면활성제와 소수성 계면활성제는 체내에서 마이셀을 형성하여 체액 내에 분포하게 된다. Hydrophilic mesoglycans are adsorbed to hydrophilic surfactants and hydrophobic choline acid is adsorbed to hydrophobic surfactants. In addition, hydrophilic and hydrophobic surfactants form micelles in the body and are distributed in the body fluid.
친수성 물질로 인해 위장관 점막의 흡수가 어려웠던 메소글리칸의 경우 본 발명의 메소클리칸 약물 제형으로 전환시 메소글리칸에 흡착된 소수성 콜린산과 계면활성제의 영향으로 소수성 콜린산과 위장관 점막간의 친화력 증진을 통해 흡수된다. 흡수된 메소글리칸 약물전달 제형은 혈액 내에서 메소글리칸과 콜린산은 친수성 및 소수성 계면활성제와 흡착되어 계면활성제의 영향으로 혈액 내에 용이하게 용해되는 것이다. In the case of mesoglycan, which was difficult to absorb gastrointestinal mucosa due to hydrophilic substances, the affinity between hydrophobic choline acid and gastrointestinal mucosa was increased due to the effect of hydrophobic choline acid and surfactant adsorbed on mesoglycan when switching to the mesoglycan drug formulation of the present invention. Is absorbed. In absorbed mesoglycan drug delivery formulations, mesoglycans and choline acids in the blood are adsorbed with hydrophilic and hydrophobic surfactants and are readily dissolved in the blood under the influence of surfactants.
또한 메소글리칸과 콜린산은 혈액 내에서 계면활성제와 흡착되어 용해 분산되어 있으므로 흡착된 계면활성제의 영향으로 메소글리칸의 지속적 방출이 가능하며 또한 메소글리칸과 콜린산은 모두 신장을 통해 용이하게 배설될 수 있다. In addition, since mesoglycans and choline acids are adsorbed and dispersed in the blood with surfactants, the mesoglycans can be continuously released under the influence of the adsorbed surfactant, and both mesoglycans and choline acids can be easily excreted through the kidneys. Can be.
따라서 본 발명의 메소글리칸 약물전달 제형은 종래 개발된 상기 식 Ⅰ의 전달제의 체내 축적에 따른 부작용을 해소할 수 있으며 또한 헤파린 결합체의 경우와 같은 의약품 약물 전달제로서의 안전성 문제를 역시 해소할 수 있는 안전한 제형이다. Therefore, the mesoglycan drug delivery formulation of the present invention can solve the side effects of the conventionally developed delivery agent of Formula I in the body and also solve the safety problem as a drug drug delivery agent such as the case of heparin conjugates. That is a safe formulation.
이하 본 발명에 사용되는 계면활성제에 대해 더욱 상세히 설명한다. Hereinafter will be described in more detail with respect to the surfactant used in the present invention.
친수성 비이온성 계면활성제는 HLB 밸류가 12 이상인 폴리에틸렌글리콜지방산에스테르임을 특징으로 하고, 소수성 비이온성 계면활성제는 HLB 밸류가 8 이하인 폴리에틸렌글리콜의 트랜스에스테르화물(transesterification product)임을 특징으로 한다. The hydrophilic nonionic surfactant is characterized by being a polyethylene glycol fatty acid ester having an HLB value of 12 or more, and the hydrophobic nonionic surfactant is characterized by being a transesterification product of polyethylene glycol having an HLB value of 8 or less.
또한 상기 폴리에틸렌글리콜지방산에스테르는 PEG-12 라우레이트, PEG-20 라우레이트, PEG-32 라우레이트, PEG-12 올레이트, PEG-20 올레이트, PEG-32 올레이트, PEG-15 스테아레이트, PEG-40 스테아레이트, PEG-20 글리세릴라우레이트, PEG-20 글리세릴올레이트, PEG-40 글리세릴라우레이트, PEG-35 캐스터오일, PEG-40 수소화 캐스터오일, PEG-30 콜레스테롤 또는 PEG-25 파이토스테롤에서 선택된 1종 이상을 사용할 수 있다. In addition, the polyethylene glycol fatty acid ester is PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-12 oleate, PEG-20 oleate, PEG-32 oleate, PEG-15 stearate, PEG -40 stearate, PEG-20 glyceryllaurate, PEG-20 glyceryl oleate, PEG-40 glyceryl laurate, PEG-35 castor oil, PEG-40 hydrogenated castor oil, PEG-30 cholesterol or PEG-25 phyto One or more selected from sterols may be used.
또한 상기 폴리에틸렌글리콜의 트랜스에스테르화물은 PEG-5 수소화 캐스터오일, PEG-7 수소화 캐스터오일, PEG-6 콘 오일, PEG-6 아몬드 오일, PEG-6 올리브 오일 또는 PEG-6 팜 케르넬 오일에서 선택된 1종 이상을 사용할 수 있다. The transesterified product of polyethylene glycol is also selected from PEG-5 hydrogenated castor oil, PEG-7 hydrogenated castor oil, PEG-6 corn oil, PEG-6 almond oil, PEG-6 olive oil or PEG-6 palm kernel oil. 1 or more types can be used.
이하 제조실시예, 제조비교예 및 실시예를 통해 본 발명을 더욱 상세히 설명한다. 그러나 하기 실시예는 본 발명의 예시를 위한 것으로 본 발명의 범위를 하기 실시예로 한정하는 것은 아니다. Hereinafter, the present invention will be described in more detail with reference to Preparation Examples, Preparation Examples and Examples. However, the following examples are intended to illustrate the present invention and are not intended to limit the scope of the invention to the following examples.
(제조실시예 1) 메소글리칸 약물전달 제형의 제조Preparation Example 1 Preparation of Mesoglycan Drug Delivery Formulation
메소글리칸 1000mg에 200mg의 데옥시콜린산(DOCA)과 친수성 비이온성 계면활성제로서 100mg의 PEG-20 라우레이트, 소수성 비이온성 계면활성제로서 100mg의 PEG-7 수소화 캐스터오일을 첨가하여 혼합시킨 메소글리칸/계면활성제/데옥시콜린산 복합체의 메소글리칸 약물전달 제형을 제조하였다. Mesoglycan mixed with 1000 mg of mesoglycans added with 200 mg of deoxycholic acid (DOCA) and 100 mg of PEG-20 laurate as a hydrophilic nonionic surfactant and 100 mg of PEG-7 hydrogenated castor oil as a hydrophobic nonionic surfactant Mesoglycan drug delivery formulations of the Khan / surfactant / deoxycholine acid complexes were prepared.
(제조실시예 2) 메소글리칸 약물전달 제형의 제조Preparation Example 2 Preparation of Mesoglycan Drug Delivery Formulation
메소글리칸 1000mg에 200mg의 데옥시콜린산(DOCA)과 친수성 비이온성 계면활성제로서 100mg의 PEG-20 올레이트, 소수성 비이온성 계면활성제로서 100mg의 PEG-5 수소화 캐스터오일을 첨가하여 혼합시킨 메소글리칸/계면활성제/데옥시콜린산 복합체의 메소글리칸 약물전달 제형을 제조하였다. Mesoglycol mixed with 1000 mg of mesoglycans added with 200 mg of deoxycholic acid (DOCA) and 100 mg of PEG-20 oleate as a hydrophilic nonionic surfactant and 100 mg of PEG-5 hydrogenated castor oil as a hydrophobic nonionic surfactant Mesoglycan drug delivery formulations of the Khan / surfactant / deoxycholine acid complexes were prepared.
(제조비교예 1) Nα-데옥시콜릴-L-라이신-메틸에스테르(식 Ⅰ)를 사용한 메소글리칸 약물전달 제형의 제조Preparation Example 1 Preparation of Mesoglycan Drug Delivery Formulation Using N α -deoxycolyl-L-lysine-Methyl Ester (Formula I)
메소글리칸 1000mg에 300mg의 Nα-데옥시콜릴-L-라이신-메틸에스테르를 혼합시켜 대한민국 특허등록 제10-679677호에 개시된 메소글리칸 약물전달 제형을 제조하였다. A mesoglycan drug delivery formulation disclosed in Korean Patent Registration No. 10-679677 was prepared by mixing 300 mg of N α -deoxycholyl-L-lysine-methylester with 1000 mg of mesoglycans.
(제조비교예 2) 아마이드 결합을 통해 화학 결합시킨 메소글리칸 약물전달 제형의 제조Preparation Example 2 Preparation of Mesoglycan Drug Delivery Formulation Chemically Bonded through Amide Bonding
Figure PCTKR2015005638-appb-I000002
Figure PCTKR2015005638-appb-I000002
대한민국 특허등록 제10-1027161호에 개시된 방법에 따라 데옥시콜린산(DOCA) 200mg을 출발물질로 하여 디사이클로헥실카르보디이미드와 히드록시숙신이미드를 DMF 용매에 용해시켜 반응시킨 후 수득된 고형 물질에 에틸렌디아민을 DMF 용매하에서 반응시킨 후 다시 1000mg의 메소글리칸을 첨가하여 상기 반응식에 나타난 바와 같은 데옥시콜린산-에틸렌디아민-메소글리칸이 결합된 메소글리칸 약물전달 제형을 제조하였다. Solid obtained after dissolving dicyclohexylcarbodiimide and hydroxysuccinimide in DMF solvent using 200 mg of deoxycholic acid (DOCA) as a starting material according to the method disclosed in Korean Patent Registration No. 10-1027161. Ethylenediamine was reacted in the DMF solvent, and then 1000 mg of mesoglycan was added to prepare a mesoglycan drug delivery formulation in which deoxycholic acid-ethylenediamine-mesoglycan was bound as shown in the above scheme.
(실시예 1) 메소글리칸 약물전달 제형의 시험관 내 항-factor Ⅹa 및 항-factor Ⅱa 활성 측정 실험Example 1 In Vitro Anti-Factor VIIa and Anti-factor IIa Activity Measurement Experiments of Mesoglycan Drug Delivery Formulations
동물 사육실에서 1주일간 순화시킨 래트를 zoletil(동물용 마취제)로 마취시킨 후 복대정맥으로부터 채혈한 후 항응고제 0.109 M 시트르산삼나트륨과 혈액이 1:9(v/v)의 비율이 되도록 혼합하였다. 혈액은 혈소판을 제거시킨 혈장(PPP)을 분리하여 실험에 사용하였다. 항-factor Ⅹa 활성 측정은 분리된 혈장(PPP)을 정제수에 일정한 농도(최종농도 0, 4, 8, 12, 16 ㎍/ml)로 용해된 제조실시예 및 제조비교예에서 제조된 약물전달 제형을 4:1의 비율로 혼합시키고 혈액응고분석기를 이용하여 표준곡선을 작성한 후 항-factor Ⅹa 활성(U/mg)을 측정하였다.Rats purified for one week in an animal feeding room were anesthetized with zoletil (animal anesthetic), collected from the vena cava, and mixed with an anticoagulant, 0.109 M trisodium citrate, to a ratio of 1: 9 (v / v). Blood was used for the experiment by separating the platelet-free plasma (PPP). The anti-factor VIIa activity was measured in the drug delivery formulations prepared in the preparation examples and the comparative examples in which isolated plasma (PPP) was dissolved in purified water at a constant concentration (final concentration 0, 4, 8, 12, 16 μg / ml). Were mixed at a ratio of 4: 1, the standard curve was prepared using a coagulation analyzer, and anti-factor VIIa activity (U / mg) was measured.
항-factor Ⅱa 활성 측정은 분리된 혈장(PPP)과 정제수에 일정한 농도(최종농도 0, 4, 8, 12, 16 ㎍/㎖)로 용해한 제조실시예 및 제조비교예에서 제조된 약물전달 제형 혼합한 후 pH 8.4 완충액 800㎕과 항 트롬빈(1IU/ml) 100㎕를 넣어 검액을 만들어 이중 400㎕를 37℃에서 4분간 가온한 후 factor Ⅱa(5U/ml) 200㎕를 넣고 잘 섞어 37℃에서 정확하게 30초간 가온한 다음 미리 37℃에서 가온 한 기질액 S-2238(1.25mmol/L) 400㎕를 넣어 잘 혼합시킨다.The anti-factor IIa activity was measured by mixing the drug delivery formulations prepared in the preparation examples and the comparative examples, which were dissolved in separated plasma (PPP) and purified water at constant concentrations (final concentrations 0, 4, 8, 12, 16 μg / ml). Then, add 800 µl of pH 8.4 buffer and 100 µl of anti thrombin (1IU / ml) to make a sample solution, and heat 400 µl of this at 37 ° C for 4 minutes, add 200 µl of factor IIa (5U / ml), and mix well at 37 ° C. After precisely warming for 30 seconds, add 400 μl of substrate solution S-2238 (1.25 mmol / L), which was warmed at 37 ° C., and mix well.
이를 37℃에서 3분간 가온한 다음 반응 정지액(50% 아세트산) 600㎕를 넣어 시험을 종료하였다. 이들 액은 마이크로플레이트 검출기를 이용하여 파장 405nm에서의 흡광도를 측정하여 제조실시예 및 제조비교예에서 제조된 약물전달 제형에 대한 표준곡선을 작성한 후 항-factor Ⅱa의 활성(U/mg)을 측정하였다.The mixture was warmed at 37 ° C. for 3 minutes, and 600 µl of the reaction stopper (50% acetic acid) was added to terminate the test. These solutions were prepared by measuring the absorbance at wavelength 405 nm using a microplate detector to prepare a standard curve for the drug delivery formulations prepared in Examples and Comparative Examples, and then measured the activity (U / mg) of anti-factor IIa. It was.
표 1은 약물전달 제형의 시험관 내 항-factor Ⅱa 및 항-factor Ⅹa의 효과를 측정한 결과를 나타낸 것이다. 제조실시예 1 및 제조실시예 2에서 제조된 약물전달 제형이 상대적으로 우수한 효과를 나타냄을 확인하였다. 또한 표준곡선을 이용하여 고분자 mg당 anti factor Ⅹa의 활성을 U(unit)로 계산하였다.Table 1 shows the results of measuring the effects of in vitro anti-factor IIa and anti-factor VIIa of the drug delivery formulation. It was confirmed that the drug delivery formulations prepared in Preparation Example 1 and Preparation Example 2 showed a relatively good effect. In addition, the activity of anti factor Ⅹa per mg of polymer was calculated as U (unit) using the standard curve.
표 1
항-factor Ⅱa 활성 항-factor Ⅹa 활성
제조실시예 1 49.13 46.24
제조실시예 2 48.79 47.37
제조비교예 1 43.40 40.54
제조비교예 2 44.22 43.11
Table 1
Anti-factor IIa activity Anti-factor VIIa activity
Preparation Example 1 49.13 46.24
Preparation Example 2 48.79 47.37
Comparative Example 1 43.40 40.54
Comparative Example 2 44.22 43.11
(단위 : U/mg)                                                     (Unit: U / mg)
실시예 1의 시험관 내 시험을 통해 메소글리칸에 친수성 계면활성제 및 소수성 계면활성제를 첨가 혼합시킨 메소글리칸 약물전달 제형이 종래에 개시된 제조비교예 1, 2에서 제조된 메소글리칸 약물전달 제형보다 우수한 항-factor Ⅱa 및 항-factor Ⅹa 효과를 나타내었다. Mesoglycan drug delivery formulations in which the hydrophilic and hydrophobic surfactants were added to and mixed with mesoglycans in the in vitro test of Example 1 were compared to those of the mesoglycan drug delivery formulations prepared in Preparation Examples 1 and 2 described above. It showed excellent anti-factor IIa and anti-factor VIIa effects.
한편 메소글리칸 및 계면활성제에 어떠한 종류의 소수성 고분자를 혼합 흡착시키는 것이 가장 우수한 항-응고 효과를 나타내는지를 측정하기 위해 콜린산, 데옥시콜린산(DOCA) 및 타우로콜린산 등 3종의 소수성 고분자를 사용하여 메소글리칸 약물전달 제형을 제조하고 그 활성을 시험관 내 및 생체 내에서 측정하였다. On the other hand, three kinds of hydrophobic compounds such as choline acid, deoxycholic acid (DOCA) and taurocholine acid were used to determine what kind of hydrophobic polymer mixed and adsorbed to mesoglycan and surfactant showed the best anti-coagulation effect. Mesoglycan drug delivery formulations were prepared using polymers and their activity was measured in vitro and in vivo.
(제조실시예 3) 메소글리칸 약물전달 제형의 제조(콜린산)Preparation Example 3 Preparation of Mesoglycan Drug Delivery Formulation (Choline Acid)
메소글리칸 1000mg에 200mg의 콜린산과 친수성 비이온성 계면활성제로서 100mg의 PEG-20 라우레이트, 소수성 비이온성 계면활성제로서 100mg의 PEG-7 수소화 캐스터오일을 첨가하여 혼합시킨 메소글리칸/계면활성제/콜린산 복합체의 메소글리칸 약물전달 제형을 제조하였다. Mesoglycan / surfactant / choline mixed with 1000 mg of mesoglycan added with 200 mg choline acid and 100 mg PEG-20 laurate as hydrophilic nonionic surfactant and 100 mg PEG-7 hydrogenated castor oil as hydrophobic nonionic surfactant Mesoglycan drug delivery formulations of the acid complex were prepared.
(제조실시예 4) 메소글리칸 약물전달 제형의 제조(콜린산)Preparation Example 4 Preparation of Mesoglycan Drug Delivery Formulation (Choline Acid)
메소글리칸 1000mg에 200mg의 콜린산과 친수성 비이온성 계면활성제로서 100mg의 PEG-20 올레이트, 소수성 비이온성 계면활성제로서 100mg의 PEG-5 수소화 캐스터오일을 첨가하여 혼합시킨 메소글리칸/계면활성제/콜린산 복합체의 메소글리칸 약물전달 제형을 제조하였다. Mesoglycan / surfactant / choline mixed with 1000 mg of mesoglycan added with 200 mg choline acid and 100 mg PEG-20 oleate as hydrophilic nonionic surfactant and 100 mg PEG-5 hydrogenated castor oil as hydrophobic nonionic surfactant Mesoglycan drug delivery formulations of the acid complex were prepared.
(제조비교예 3) 메소글리칸 약물전달 제형의 제조(데옥시콜린산)Preparation Example 3 Preparation of Mesoglycan Drug Delivery Formulation (Deoxycholic Acid)
메소글리칸 1000mg에 200mg의 데옥시콜린산(DOCA)과 친수성 비이온성 계면활성제로서 100mg의 PEG-20 라우레이트, 소수성 비이온성 계면활성제로서 100mg의 PEG-7 수소화 캐스터오일을 첨가하여 혼합시킨 메소글리칸/계면활성제/데옥시콜린산 복합체의 메소글리칸 약물전달 제형을 제조하였다. Mesoglycan mixed with 1000 mg of mesoglycans added with 200 mg of deoxycholic acid (DOCA) and 100 mg of PEG-20 laurate as a hydrophilic nonionic surfactant and 100 mg of PEG-7 hydrogenated castor oil as a hydrophobic nonionic surfactant Mesoglycan drug delivery formulations of the Khan / surfactant / deoxycholine acid complexes were prepared.
(제조비교예 4) 메소글리칸 약물전달 제형의 제조(타우로콜린산)Preparation Example 4 Preparation of Mesoglycan Drug Delivery Formulation (Taurocholic Acid)
메소글리칸 1000mg에 200mg의 타우로콜린산과 친수성 비이온성 계면활성제로서 100mg의 PEG-20 라우레이트, 소수성 비이온성 계면활성제로서 100mg의 PEG-7 수소화 캐스터오일을 첨가하여 혼합시킨 메소글리칸/계면활성제/타우로콜린산 복합체의 메소글리칸 약물전달 제형을 제조하였다. Mesoglycan / surfactant mixed with 1000 mg of mesoglycan added with 200 mg of taurocholic acid and 100 mg of PEG-20 laurate as a hydrophilic nonionic surfactant and 100 mg of PEG-7 hydrogenated castor oil as a hydrophobic nonionic surfactant Mesoglycan drug delivery formulations of the / taurocholine acid complex were prepared.
(실시예 2) 메소글리칸 약물전달 제형의 생체 내 항-factor Ⅹa 및 항-factor Ⅱa 활성 측정 실험Example 2 In Vivo Anti-Factor VIIa and Anti-factor IIa Activity Measurement Experiments of Mesoglycan Drug Delivery Formulations
동물 사육실에서 1주일간 순화시킨 래트에 제조실시예 3, 4 및 제조비교예 3, 4에서 제조된 메소글리칸 약물전달 제형을 물에 용해, 현탁시켜 경구 투여하였다. 복대 정맥 및 안와 정맥에서 경구 투여 후 15분, 30분, 1시간, 5시간 채혈하여 항응고제 0.109 M 시트르산삼나트륨과 혈액이 1:9(v/v)의 비율이 되도록 혼합하였다. 혈액은 혈소판을 제거시킨 혈장(PPP)을 분리하여 실험에 사용하였다. 원심 분리하여 얻은 혈소판을 제거시킨 혈장(PPP)에 대한 항-factor Ⅹa 활성은 Hemosil liquid heparin kit을 이용하였다.The mesoglycan drug delivery formulations prepared in Examples 3 and 4 and Comparative Examples 3 and 4 were dissolved or suspended in water and orally administered to rats purified for one week in an animal rearing room. Blood samples were collected for 15 minutes, 30 minutes, 1 hour, and 5 hours after oral administration in the abdominal and orbital veins, and the anticoagulant 0.109 M trisodium citrate was mixed with blood at a ratio of 1: 9 (v / v). Blood was used for the experiment by separating the platelet-free plasma (PPP). Hemosil liquid heparin kit was used for anti-factor VIIa activity against plasma (PPP) from which platelets obtained by centrifugation were removed.
항-factor Ⅱa 활성 측정은 원심 분리하여 얻은 혈소판을 제거시킨 혈장(PPP) 100㎕에 pH 8.4 완충액 800㎕과 항 트롬빈(1IU/ml) 100㎕를 넣어 검액으로 하였다. 검액을 제외한 헤파린 표준액 및 실험방법은 시험관 내 실험 항-factor Ⅱa 활성 측정과 동일하게 진행하였다.Anti-factor IIa activity was measured by adding 800 μl of pH 8.4 buffer and 100 μl of antithrombin (1IU / ml) to 100 μl of platelet-free plasma (PPP) obtained by centrifugation. Heparin standard solution and test method except the test solution were performed in the same manner as the in vitro experimental anti-factor IIa activity measurement.
표 2는 제조실시예 3, 4 및 제조비교예 3, 4에서 제조된 메소글리칸 약물전달 제형을 래트에 경구 투여하여 15분, 30분, 1시간, 5시간 후 혈장에서의 항-factor Ⅹa 활성을 측정하였다. 그 결과로 제조실시예 3, 4에서 제조된 메소글리칸 약물전달 제형은 30분에서 가장 높게 관찰되었고 반면 제조비교예 3에서 제조된 메소글리칸 약물전달 제형은 60분에서 높게 관찰되었으나 제조비교예 4에서 제조된 메소글리칸 약물전달 제형은 생체 내에서 거의 효과를 나타내지 못하였다.Table 2 shows the anti-factor VIIa in plasma after 15 minutes, 30 minutes, 1 hour, and 5 hours by oral administration of the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 and Comparative Examples 3 and 4 to rats. Activity was measured. As a result, the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 were highest at 30 minutes, whereas the mesoglycan drug delivery formulations prepared in Preparation Example 3 were observed at 60 minutes, but the preparation comparison Mesoglycan drug delivery formulations prepared in 4 showed little effect in vivo.
한편 제조실시예 3에서 제조된 메소글리칸 약물전달 제형은 30분에서 가장 높은 항-factor Ⅹa 활성인 0.72U/ml이 관찰되었으며 제조실시예 4에서 제조된 메소글리칸 약물전달 제형은 30분에서 가장 높은 항-factor Ⅹa 활성인 0.68U/ml이 관찰되었다. Meanwhile, the mesoglycan drug delivery formulation prepared in Preparation Example 3 showed the highest anti-factor VIIa activity of 0.72 U / ml at 30 minutes, and the mesoglycan drug delivery formulation prepared in Preparation Example 4 was used at 30 minutes. The highest anti-factor VIIa activity of 0.68 U / ml was observed.
표 2 항-factor Ⅹa 활성
15분(U/ml) 30분(U/ml) 60분(U/ml) 300분(U/ml)
제조실시예 3 0.48 0.72 0.52 0.17
제조실시예 4 0.49 0.68 0.54 0.19
제조비교예 3 0.30 0.37 0.44 0.08
제조비교예 4 0.10 0.14 0.07 0.01
TABLE 2 Anti-factor VIIa activity
15 minutes (U / ml) 30 minutes (U / ml) 60 minutes (U / ml) 300 minutes (U / ml)
Preparation Example 3 0.48 0.72 0.52 0.17
Preparation Example 4 0.49 0.68 0.54 0.19
Comparative Example 3 0.30 0.37 0.44 0.08
Comparative Example 4 0.10 0.14 0.07 0.01
표 3은 제조실시예 3, 4 및 제조비교예 3, 4에서 제조된 메소글리칸 약물전달 제형을 래트에 경구 투여하여 15분, 30분, 1시간, 5시간 후 혈장에서의 항-factor Ⅱa 활성을 측정하였다. 그 결과로 제조실시예 3, 4에서 제조된 메소글리칸 약물전달 제형은 30분에서 가장 높게 관찰되었고 반면 제조비교예 3에서 제조된 메소글리칸 약물전달 제형은 60분에서 높게 관찰되었으나 제조비교예 4에서 제조된 메소글리칸 약물전달 제형은 생체 내에서 거의 효과를 나타내지 못하였다.Table 3 shows the anti-factor IIa in plasma after 15 minutes, 30 minutes, 1 hour, and 5 hours by oral administration of the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 and Comparative Examples 3 and 4. Activity was measured. As a result, the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 were highest at 30 minutes, whereas the mesoglycan drug delivery formulations prepared in Preparation Example 3 were observed at 60 minutes, but the preparation comparison Mesoglycan drug delivery formulations prepared in 4 showed little effect in vivo.
한편 제조실시예 3에서 제조된 메소글리칸 약물전달 제형은 30분에서 가장 높은 항-factor Ⅱa 활성인 0.34U/ml이 관찰되었으며 제조실시예 4에서 제조된 메소글리칸 약물전달 제형은 30분에서 가장 높은 항-factor Ⅱa 활성인 0.35U/ml이 관찰되었다. Meanwhile, the mesoglycan drug delivery formulation prepared in Preparation Example 3 showed the highest anti-factor IIa activity at 0.34 U / ml at 30 minutes, and the mesoglycan drug delivery formulation prepared at Preparation Example 4 was used at 30 minutes. The highest anti-factor IIa activity, 0.35 U / ml, was observed.
표 3 항-factor Ⅱa 활성
15분(U/ml) 30분(U/ml) 60분(U/ml) 300분(U/ml)
제조실시예 3 0.25 0.34 0.32 0.08
제조실시예 4 0.21 0.35 0.27 0.10
제조비교예 3 0.10 0.14 0.20 0.00
제조비교예 4 0.05 0.07 0.02 0.00
TABLE 3 Anti-factor IIa activity
15 minutes (U / ml) 30 minutes (U / ml) 60 minutes (U / ml) 300 minutes (U / ml)
Preparation Example 3 0.25 0.34 0.32 0.08
Preparation Example 4 0.21 0.35 0.27 0.10
Comparative Example 3 0.10 0.14 0.20 0.00
Comparative Example 4 0.05 0.07 0.02 0.00
(실시예 3) 시험관 내 활성화 부분 트롬보플라스틴 시간(APTT)의 측정 Example 3 Measurement of In Vitro Activated Partial Thromboplastin Time (APTT)
동물 사육실에서 1주일간 순화시킨 래트에 제조실시예 3, 4 및 제조비교예 3, 4에서 제조된 메소글리칸 약물전달 제형을 물에 용해, 현탁시켜 경구 투여하였다. 다음으로 복대정맥 및 안와정맥으로부터 경구 투여 후 15분, 30분, 1시간, 5시간 채혈하여 항응고제 0.109M 시트르산삼나트륨과 혈액이 1:9(v/v)의 비율이 되도록 혼합하였다. 혈액은 혈소판을 제거시킨 혈장(PPP)을 분리하여 실험에 사용하였다. 원심 분리하여 얻은 혈소판을 제거시킨 혈장(PPP)에 대한 APTT는 ACL-에세이 시약 키트를 이용하여 자동혈액응고분석기(ACL-7000)로 측정하였다.The mesoglycan drug delivery formulations prepared in Examples 3 and 4 and Comparative Examples 3 and 4 were dissolved or suspended in water and orally administered to rats purified for one week in an animal rearing room. Next, blood was collected for 15 minutes, 30 minutes, 1 hour, and 5 hours after oral administration from the abdominal and orbital veins, and the anticoagulant 0.109M trisodium citrate and blood were mixed at a ratio of 1: 9 (v / v). Blood was used for the experiment by separating the platelet-free plasma (PPP). APTT for platelet-depleted plasma (PPP) obtained by centrifugation was measured by an automated blood coagulation analyzer (ACL-7000) using an ACL-assay reagent kit.
표 4는 제조실시예 3, 4 및 제조비교예 3, 4에서 제조된 메소글리칸 약물전달 제형을 경구 투여하여 15분, 30분, 1시간, 5시간 후 혈장에서의 APTT를 측정한 결과이다. 제조실시예 3에서 제조된 메소글리칸 약물전달 제형의 경우 30분에서 46.3초와 1시간에서 57.5초로 APTT를 연장시킨 것으로 관찰되었다Table 4 shows the results of measuring APTT in plasma after 15 minutes, 30 minutes, 1 hour and 5 hours by oral administration of the mesoglycan drug delivery formulations prepared in Preparation Examples 3 and 4 and Comparative Examples 3 and 4. . In the mesoglycan drug delivery formulation prepared in Preparation Example 3, it was observed that APTT was extended from 30 minutes to 46.3 seconds and from 1 hour to 57.5 seconds.
표 4 활성화 부분 트롬보플라스틴 시간(APTT)
15분 30분 60분 300분
정상(normal) 17.6 - - -
제조실시예 3 29.4 46.3 57.5 27.4
제조실시예 4 28.2 44.7 50.5 20.6
제조비교예 3 17.4 24.7 19.4 15.7
제조비교예 4 16.9 19.1 17.5 14.2
Table 4 Activation partial thromboplastin time (APTT)
15 minutes 30 minutes 60 minutes 300 minutes
Normal 17.6 - - -
Preparation Example 3 29.4 46.3 57.5 27.4
Preparation Example 4 28.2 44.7 50.5 20.6
Comparative Example 3 17.4 24.7 19.4 15.7
Comparative Example 4 16.9 19.1 17.5 14.2
(단위 : 초)                                                       (Unit: seconds)

Claims (5)

1) 100 중량부의 메소글리칸, 2) 10~50 중량부의 콜린산 및 3) 10~100 중량부의 친수성 및 소수성 계면활성제를 흡착 결합시킨 메소글리칸 약물 전달 제형에 있어서, 상기 친수성 계면활성제는 HLB 밸류가 12 이상인 비이온성 계면활성제이고, 상기 소수성 계면활성제는 HLB 밸류가 8 이하인 비이온성 계면활성제임을 특징으로 하는 메소글리칸 약물전달 제형.In a mesoglycan drug delivery formulation comprising 1) 100 parts by weight of mesoglycans, 2) 10 to 50 parts by weight of choline acid and 3) 10 to 100 parts by weight of hydrophilic and hydrophobic surfactants, the hydrophilic surfactant is HLB. Mesoglycan drug delivery formulation, characterized in that the nonionic surfactant having a value of 12 or more, the hydrophobic surfactant is a nonionic surfactant having an HLB value of 8 or less.
제 1항에 있어서, 상기 친수성 계면활성제와 상기 소수성 계면활성제의 함량은 중량비로 0.8~1.2 : 1.2~0.8 임을 특징으로 하는 메소글리칸 약물전달 제형.The mesoglycan drug delivery formulation of claim 1, wherein the hydrophilic surfactant and the hydrophobic surfactant are 0.8 to 1.2: 1.2 to 0.8 by weight.
제 1항 또는 제 2항에 있어서, 상기 친수성 비이온성 계면활성제는 HLB 밸류가 12 이상인 폴리에틸렌글리콜지방산에스테르이며, 상기 소수성 비이온성 계면활성제는 HLB 밸류가 8 이하인 폴리에틸렌글리콜의 트랜스에스테르화물(transesterification product)임을 특징으로 하는 메소글리칸 약물전달 제형.3. The transesterification product of polyethylene glycol according to claim 1 or 2, wherein the hydrophilic nonionic surfactant is polyethylene glycol fatty acid ester having an HLB value of 12 or more, and the hydrophobic nonionic surfactant is a transesterification product of polyethylene glycol having an HLB value of 8 or less. Mesoglycan drug delivery formulation characterized in that.
제 3항에 있어서, 상기 폴리에틸렌글리콜지방산에스테르는 PEG-12 라우레이트, PEG-20 라우레이트, PEG-32 라우레이트, PEG-12 올레이트, PEG-20 올레이트, PEG-32 올레이트, PEG-15 스테아레이트, PEG-40 스테아레이트, PEG-20 글리세릴라우레이트, PEG-20 글리세릴올레이트, PEG-40 글리세릴라우레이트, PEG-35 캐스터오일, PEG-40 수소화 캐스터오일, PEG-30 콜레스테롤 또는 PEG-25 파이토스테롤에서 선택된 1종 이상임을 특징으로 하는 메소글리칸 약물전달 제형.The method of claim 3, wherein the polyethylene glycol fatty acid ester is PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-12 oleate, PEG-20 oleate, PEG-32 oleate, PEG- 15 stearate, PEG-40 stearate, PEG-20 glyceryl laurate, PEG-20 glyceryl oleate, PEG-40 glyceryl laurate, PEG-35 castor oil, PEG-40 hydrogenated castor oil, PEG-30 cholesterol Or PEG-25 phytosterols; at least one mesoglycan drug delivery formulation.
제 3항에 있어서, 상기 폴리에틸렌글리콜의 트랜스에스테르화물은 PEG-5 수소화 캐스터오일, PEG-7 수소화 캐스터오일, PEG-6 콘 오일, PEG-6 아몬드 오일, PEG-6 올리브 오일 또는 PEG-6 팜 케르넬 오일에서 선택된 1종 이상임을 특징으로 하는 메소글리칸 약물전달 제형.The method of claim 3, wherein the transesterified polyethylene glycol is PEG-5 hydrogenated castor oil, PEG-7 hydrogenated castor oil, PEG-6 corn oil, PEG-6 almond oil, PEG-6 olive oil or PEG-6 palm Mesoglycan drug delivery formulation, characterized in that at least one selected from Kernel oil.
PCT/KR2015/005638 2014-10-07 2015-06-05 Improved mesoglycan drug delivery dosage form WO2016056728A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020140134739A KR101528476B1 (en) 2014-10-07 2014-10-07 Enhanced drug delivery formulation for mesoglycan
KR10-2014-0134739 2014-10-07

Publications (1)

Publication Number Publication Date
WO2016056728A1 true WO2016056728A1 (en) 2016-04-14

Family

ID=53503718

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2015/005638 WO2016056728A1 (en) 2014-10-07 2015-06-05 Improved mesoglycan drug delivery dosage form

Country Status (2)

Country Link
KR (1) KR101528476B1 (en)
WO (1) WO2016056728A1 (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20010024658A1 (en) * 1999-08-17 2001-09-27 Feng-Jing Chen Pharmaceutical dosage form for oral administration of hydrophilic drugs, particularly low molecular weight heparin
KR100679677B1 (en) * 2004-05-21 2007-02-07 주식회사 메디프렉스 Delivery Agents for Enhancing Mucosal Absorption of Biologically Active Agents
KR100891278B1 (en) * 2004-02-12 2009-04-06 한국과학기술연구원 Composition and formulation of colloidal system comprising biocompatible aqueous-soluble polymer, and preparation method thereof
WO2011003045A1 (en) * 2009-07-01 2011-01-06 Magceutics, Inc. Slow release magnesium composition and uses thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20010024658A1 (en) * 1999-08-17 2001-09-27 Feng-Jing Chen Pharmaceutical dosage form for oral administration of hydrophilic drugs, particularly low molecular weight heparin
KR100891278B1 (en) * 2004-02-12 2009-04-06 한국과학기술연구원 Composition and formulation of colloidal system comprising biocompatible aqueous-soluble polymer, and preparation method thereof
KR100679677B1 (en) * 2004-05-21 2007-02-07 주식회사 메디프렉스 Delivery Agents for Enhancing Mucosal Absorption of Biologically Active Agents
WO2011003045A1 (en) * 2009-07-01 2011-01-06 Magceutics, Inc. Slow release magnesium composition and uses thereof

Also Published As

Publication number Publication date
KR101528476B1 (en) 2015-06-12

Similar Documents

Publication Publication Date Title
WO2011081430A2 (en) Polymer nanoparticle injection formulation composition containing rapamycin with improved water solubility, preparation method thereof, and anticancer composition for combined use with radiotherapy
KR20020083905A (en) Oral Delivery of Macromolecules
ES2568455T3 (en) Compounds for the treatment of Clostridium difficile-associated disease
WO2006102717A1 (en) Modulating angiogenesis with nod factors such as glucosamine oligosaccharides
EP0658585B1 (en) Aminophenylalanine derivatives, process for their production, their use and agents containing them as anti-coagulants
WO2016056728A1 (en) Improved mesoglycan drug delivery dosage form
WO2022019701A1 (en) Anti-adhesion polymer composition
WO2023158092A1 (en) Pharmaceutical compositions for hemostasis and wound healing in gastrointestinal tract
WO2022211493A1 (en) Pectin modified with gallol derivative and use thereof
WO2017014332A1 (en) Novel drug delivery conjugated moiety for oral administration of drug unsuitable for oral administration and preparation method thereof
EP3938376A1 (en) Novel peptide and use thereof
WO2013176455A1 (en) Multilayer coating form of orally administered pharmaceutical composition containing omega-3 fatty acid or alkyl ester thereof and statin based drug
WO2022131820A1 (en) Anti-inflammatory peptide for preventing or treating atopic dermatitis
WO2022114881A1 (en) Pharmaceutical composition for preventing or treating wound or scar, comprising benzbromarone
CN112409439B (en) Glycyrrhizic acid derivative, preparation method and application
WO2010074460A2 (en) Novel pharmaceutical composition for ameliorating side effects of a lipase inhibitor
WO2021107427A1 (en) Cosmetic composition for skin barrier enhancement or skin bioactive substance absorption promotion
WO2016064202A1 (en) Thermosensitive pluronic-chitosan oligosaccharide-kartogenin nanocomposite with dual drug release function
WO2020222552A1 (en) Pharmaceutical composition, comprising 6-diazo-5-oxo-l-norleucine, for treatment of inflammatory skin disease
WO2011122899A2 (en) Pharmaceutical composition for inhibiting platelet aggregation or for thrombolysis
WO2016163752A1 (en) Soluble microneedle for delivering poorly-soluble drug
WO2011031042A2 (en) Pegylated triiodinated aryl compound, and x-ray contrast agent comprising same
WO2019139313A1 (en) Stabilized pharmaceutical formulation comprising everolimus
WO2023234747A1 (en) Pectin modified with phenolic derivative and use thereof
WO2021251565A1 (en) Pharmaceutical composition comprising benzimidazole derivative compound

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15849420

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15849420

Country of ref document: EP

Kind code of ref document: A1