WO2014069846A1 - Method for preparing 1-deoxy-1-(2-hydroxyethyl amino)-d-glucitol and miglitol - Google Patents

Method for preparing 1-deoxy-1-(2-hydroxyethyl amino)-d-glucitol and miglitol Download PDF

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WO2014069846A1
WO2014069846A1 PCT/KR2013/009599 KR2013009599W WO2014069846A1 WO 2014069846 A1 WO2014069846 A1 WO 2014069846A1 KR 2013009599 W KR2013009599 W KR 2013009599W WO 2014069846 A1 WO2014069846 A1 WO 2014069846A1
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deoxy
hydroxyethyl amino
present
ammonium
organic solvent
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French (fr)
Korean (ko)
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서정우
함윤범
강흥모
박지수
이광무
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종근당바이오 주식회사
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C213/00—Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton
    • C07C213/10—Separation; Purification; Stabilisation; Use of additives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C213/00—Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton
    • C07C213/02—Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton by reactions involving the formation of amino groups from compounds containing hydroxy groups or etherified or esterified hydroxy groups
    • 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
    • A61K31/13—Amines
    • A61K31/133—Amines having hydroxy groups, e.g. sphingosine
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C215/00—Compounds containing amino and hydroxy groups bound to the same carbon skeleton
    • C07C215/02—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C215/04—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being saturated
    • C07C215/06—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being saturated and acyclic
    • C07C215/12—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being saturated and acyclic the nitrogen atom of the amino group being further bound to hydrocarbon groups substituted by hydroxy groups

Definitions

  • the present invention provides a process for preparing 1-deoxy-1— (2-hydroxyethylamino) ⁇ D—glucile from agglutination using a borohydride complex as a reducing agent and the 1-deoxy-1- It relates to a process for producing migli from (2-hydroxyethyl amino) -D-glucos.
  • Miglyul is an ⁇ -glucosidase inhibitor developed by Bayer and obtained FDA approval in 1996 and is currently sold under the trade name Glyset for the treatment of crab type 2 diabetes. It is becoming.
  • the manufacturing process of Miglet is the first step in synthesizing 1-deoxy-1- (2-hydroxyethyl amino) -D-glucose from glucose and ethane from amines, as shown in [Bannung 2]. Is synthesized through the second step of bioconversion to 6-hydroxyethyl amino-L-sorbose using the biocatalyst and the third step of synthesizing Migly through chemical synthesis ra / Jcei / Materials Research , 197-198: p. 51-55 (2011)).
  • Migly is prepared from 1-deoxy— 1- (2-hydroxyethyl amino) -D-glusy to 6-hydroxyethyl amino-L-sorbose specified above: It can be seen that the reaction is an important process, and the method for preparing 1-deoxy— 1- (2-hydroxyethyl amino) -D-glucit, an intermediate required for this bioconversion reaction, is shown in [Scheme 4]. There is a hydrogenation reaction using palladium-on-charcoal (EP0447160).
  • the present inventors can produce in high yield 1-deoxy-1- (2-hydroxyethyl amino) -D-glusy, which is an intermediate for preparing Migly. Efforts have been made to develop new manufacturing methods which have been able to ameliorate the problems of the production process risks and low yields. As a result, when the borohydride complex is used as a reducing agent in an organic solvent containing ethanolamine, the present invention was completed by confirming that the intermediates can be produced in high yield without the risk of explosion.
  • Another object of the present invention is to provide a method for producing Migly.
  • the present invention is a ethanol under an organic solvent containing an amine, using the borohydride complex as a reducing agent to the D-glucin 1-deoxy -1- (2-hydride of formula 1
  • a process for preparing 1-deoxy-1- (2'hydroxyethyl amino) -D-glucy characterized in that oxyethyl amino) -D-glucos is prepared by:
  • the inventors of the present invention can produce 1-deoxy-1 '(2-hydroxyethyl amino) -D-glusy, which is an intermediate for preparing Migly, in high yield, and the production process of the conventional manufacturing method as described above. Efforts have been made to develop new methods of manufacturing that could improve the risks of phase and problems of low yield. As a result, it was confirmed that when the borohydride complex was used as a reducing agent in an organic solvent containing ethane, amines could be produced in high yield without a risk of explosion of the reaction.
  • the biggest feature of the present invention is the use of palladium-ion-charcoal for the hydrogenation reaction with the risk of explosion in the preparation of 1-deoxy— 1- (2-hydroxyethyl amino) -D-glue.
  • the borohydride complex is used as a reducing agent.
  • the preparation method of the present invention is shown in the following reaction formula 1.
  • the preparation method of the present invention using the borohydride complex is 1-deoxy-1- (2 ⁇ hydroxyethyl amino) -D compared to the preparation method using the conventional palladium-on-charcoal. -Significantly increase the production yield of glue (see Table 1).
  • the production yield of 1-deoxysulf 1- (2-hydroxyethyl amino) -euglucin according to the preparation method of the present invention is 70-
  • the ' borohydride complex is selected from the group consisting of sodium borohydride (NaB3 ⁇ 4), sodium borohydride (NaCNB3 ⁇ 4) and dimethylaminoborane ( BH 4 N ( C H 3) 2) .
  • the borohydride complex is sodium borohydride or sodium borocyanohydride, and in another specific example, sodium borohydride.
  • the amount of boron hydride used is 1 to 10 equivalents relative to the number of moles of D-glucose, in one particular example 2 to 5 equivalents, and in another specific example 2 to 3 equivalents.
  • the organic solvent includes an ammonium salt.
  • the ammonium salt is ammonium acetate (C3 ⁇ 4C00NH 4 ), ammonium monophosphate (NH 4 H 2 P0 4 ), ammonium diphosphate ((NH 4 ) 2 HP0 4 ), ammonium sulfate ((NH 4 ) 2 S0 4 ), ammonium nitrate (NH 4 N0 3 ) and ammonium carbonate ((NH 4 ) 2 C0 3 ), in another specific example ammonium acetate.
  • the amount of ammonium salt is used is 1 to 20 equivalents based on D-glucose and the number of moles, 3 to 15 equivalents in one specific example, and 5 to 10 equivalents in another specific example.
  • the organic solvent is an alcohol solvent such as methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol; Ester solvents including ethyl acetate; Halo alkanes solvents including methylene chloride and chloroform; Ether solvents including tetrahydrofuran; Keron solvents including acetone and methyl ethyl ketone; Acetonitrile: nitrile solvents including propionitrile; Amide solvents including dimethylformamide and dimethylacetamide; Sulfoxide solvents including dimethyl sulfoxide; A solvent in which the two or more organic solvents are mixed; Or a solvent in which the organic solvent and water are mixed.
  • alcohol solvent such as methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol
  • Ester solvents including ethyl acetate
  • Halo alkanes solvents including methylene chloride and chloroform
  • Ether solvents including tetrahydrofuran
  • the organic solvent is an alcoholic solvent
  • the alcoholic solvent is ethyl alcohol
  • the volume of the solvent used in the preparation method of the present invention is 1: 1-20 as the volume (m «ratio) relative to the mass (g) of D-glucose, in one specific example 1: 1-15, and in another specific example 1: 5-10.
  • the production method of the present invention proceeds the reaction at a lower temperature than the conventional production method using palladium ⁇ -charcoal.
  • the reaction process of the present invention proceeds at a temperature of -20 to 60 ° C, in another specific example 0 to 4 (running at a temperature of rc, and in another specific example of 20 to 40 ° C) Proceed at temperature.
  • the reaction time is 5 minutes to 24 hours, in one specific example is 30 minutes to 5 hours, and in another specific example is 30 minutes to 1 hour.
  • the product-containing solution in each case can be concentrated by chromatography with an ion exchanger, silica gel or silylated silica gel and crystallized with a suitable solvent.
  • Suitable ion exchangers of the present invention include both weakly acidic, strongly acidic, weakly basic and strongly basic forms, and in one particular example are strongly acidic or strongly basic ion exchangers.
  • Suitable crystallization solvents of the present invention include ester solvents such as ethyl acetate; Halo al3 ⁇ 4 solvents such as methylene chloride and chloroform; Ether solvents such as tetrahydrofuran; Ketone solvents such as acetone and methyl ethyl ketone; Nitrile solvents such as acetonitrile and propionitrile; Alcohol solvents such as methyl alcohol, ethyl alcohol, propanol and isopropane; And organic solvents in which two or more thereof are mixed.
  • ester solvents such as ethyl acetate
  • Halo al3 ⁇ 4 solvents such as methylene chloride and chloroform
  • Ether solvents such as tetrahydrofuran
  • Ketone solvents such as acetone and methyl ethyl ketone
  • Nitrile solvents such as acetonitrile and propionitrile
  • Alcohol solvents such as methyl alcohol, ethyl alcohol
  • the crystallization solvent is ethyl alcohol.
  • the volume used of the crystallization solvent is from 1.0 to 20 times by volume () ratio to the weight (g) of D-glucose used, in one particular example from 5 to 10 times.
  • the progress of the reaction process can be analyzed and measured using a means such as thin layer chromatography or ELSD (Evaporative Light Scattering Detector).
  • ELSD Electrode Light Scattering Detector
  • step (b) 1-deoxy-1- (2-hydroxyethyl amino) -D-glucos prepared in step (a) using 6-glucobacterium strain or resting cells thereof. Bioconversion to hydroxyethyl amino-L-sorbose; And
  • step (c) synthesizing 6-hydroxyethyl amino-L-sorbose biomigrated in step (b) into Migly.
  • the method for producing Miglyol of the present invention uses the above-described method for preparing 1-deoxy-1- (2-hydroxyethyl amino) -D-glycine, these two The common content between them is omitted in order to avoid excessive complexity of the present specification.
  • step (b) comprises a 1-deoxy-1- (2-hydroxyethyl amino) -D-glucose of dormant cells of the bacterium Gluconobacter or the microorganism. Incubate in medium.
  • 1-deoxy-1- (2-hadoxyethylamino) -D-glucy may be added before inoculating the silver strain or after inoculation of the strain, and added at the same time as the start of the culture. Or it can be added continuously or in portions throughout the culture process.
  • the amount of 1-deoxy-1- (2-hydroxyethyl amino) -agglucius used can be determined experimentally by one skilled in the art.
  • _ 6-hydroxy-ethylamino -L- medium for the production of a sorbose is a 1-deoxy-eu 1- (2-hydroxy-ethylamino) -D- gluconic when the the 10 to 150 gl It may be contained in a concentration, in another specific example may be contained in a concentration of 100 g / ⁇ within 10 gl i, in another specific example may be contained in a concentration of 10 gl i to 80 g /.
  • the medium When the strain is used in the form of a resting cell paste, the medium contains 1-deoxy—1 ′ (2-hydroxyethyl amino) -D-glusy at a concentration of 10 g / H to 200. In another specific example, it may contain at a concentration of 10 g / i to 150.
  • the medium used in the preparation method of the present invention is not only 1-deoxy-1- (2-hydroxyethyl amino) -D-glue, but also D-glue, nitrogen source inorganic salts and trace elements, and growth factors. It may include, and may include at least one other carbon source.
  • the amount of each component added is preferably selected to maximize the production of 6-hydroxyethyl amino-L-sorbose, and the amount can be determined experimentally by one skilled in the art according to various methods known in the art. .
  • Culture conditions for maximizing the production of 1-deoxy-1- (2-hydroxyethyl amino) glue from 6-hydroxyethyl amino-L-sorbose as temperature, pH, aeration, stirring, incubation period Optimization can be determined experimentally by one skilled in the art.
  • the choice of specific culture conditions may vary depending on the strain used, dormant cells, medium composition, culture size and other considerations.
  • the culture temperature of the strain in the production method of the present invention is made at 15 to 37 ° C, in one specific example is made at 25 ° C.
  • Suitable pH of the culture is in the range from 4.0 to 8.0, in one particular range from 4.5 to 7.0, in another particular range from 5.5 to 6.0.
  • 6-hydroxyethyl amino-L-sorbose cell extract of the genus Gluconobacter strain or dormant cell extract of the microorganism may be used.
  • the method for preparing 6-hyduoxyethyl amino-L-sorbose using cell extracts is (a) cell extracts of the genus Gluconobacter or cell extracts of dormant cells of the microorganism. Contacting hydroxyethyl amino) -D-gluci to; And (b) recovering 6-hydroxyethyl amino-L-sorbose.
  • the strains of the genus Gluconobacter are gluconobacter oxydans, and in one particular example, gluconobacter oxydans CKDBM 0901.
  • the Gluconobacter Oxydans CKDBM 0901 is resistant to 1—deoxy-1- (2-hydroxyethylamino) -D-glucy and 1-deoxy-1- (2-hydroxyethyl amino)-
  • step (c) of the present invention bio-converted 6-hydroxyethyl amino-L-sorbose is synthesized in Miglit.
  • Miglyul is (2R, 3R, 4R, 5S) -l- (2-hydroxyethyl) -2- (hydroxymethyl) piper i dine— 3,4,5—trk) l, an ⁇ -glucosidase inhibitor Oral type 2 diabetes treatment.
  • the method for synthesizing migli using 6-hydroxyethyl amino-L-sorbose obtained in step (b) is 6-
  • the culture solution containing hydroxyethyl amino-L-sorbose was filtered and then treated with activated carbon to remove the pigment and transferred to a hydrogenation reactor: Migley was carried out by carrying out the synthesis.
  • Palladium "on-charcoal may be used as a catalyst for hydrogenation reaction, and reaction is performed at the temperature and pressure conditions suitable for reaction, and it removes a catalyst by filtering.
  • the mixture was eluted with a resin, eluted and concentrated, and then dried through a crystallization and recrystallization process to obtain a high purity migliol in the form of an all-white powder.
  • the present invention provides a process for preparing 1-deoxy ⁇ 1- (2-hydroxyethylamino) -D-glue from D-glucos using boron hydride complex as reducing agent, and 1 prepared -Deoxy-; L- ( 2 -hydroxyethyl amino) -D-gluclet is bioconverted to provide a method for producing migli.
  • the preparation method of the present invention uses boron hydride complexes instead of palladium-on-charcoal to yield 1-deoxy-1- (2-hydroxyethyl amino)-in high yield without the risk of explosion.
  • D can produce glue.
  • the present invention can be usefully used for the production of Migly, a therapeutic agent for diabetes.
  • Figure 1 is an NMR result for 1-deoxy-1- (2-hydroxyethyl amino) -D-glucos prepared according to the method of the present invention.
  • FIG. 2 shows the results of an Evaporative Light Scattering Detector (ELSD) for lnddeoxy-1- (2-hydroxyethyl amino) -D-glucis prepared according to the method of the present invention.
  • ELSD Evaporative Light Scattering Detector
  • FIG. 3 is a Mass result for 1-deoxy-1- (2-hydroxyethyl amino) -D-glusytle prepared according to the method of the present invention.
  • Figure 4 shows the spectrum analyzed by infrared spectroscopy for Migley prepared according to the method of the present invention.
  • 5 is an NMR result for Migly prepared according to the method of the present invention.
  • the terminated reaction solution was adsorbed onto NAC-4 strong acid cation exchanger and washed with purified water, followed by eluting 30% aqueous ammonia and methyl alcohol with eluent and concentrating. The resulting fractions were concentrated in vacuo and the residue obtained was crystallized from ethyl alcohol, filtered and dried to give 102 g (theoretical yield 82%) of 1-deoxy-1- (2 ′ hydroxyethyl amino) -D-glucy ( Purity: 99.9%).
  • the finished reaction solution was adsorbed on a NAC-4 strongly acidic cation exchanger, washed with purified water, and then eluted with 30% aqueous ammonia and methyl alcohol and concentrated. The resulting fractions were concentrated in vacuo and the residue obtained was crystallized from ethyl alcohol, filtered and dried to give 100 g of 1-deoxy-1- (2-hydroxyethyl amino) -D-glusy (803 ⁇ 4 »theoretical water). (Purity: 99.7%).
  • Table 1 shows the results of Preparation Example 1, Preparation Example 2, and Comparative Example of 1-deoxy-1- (2-hydroxyethyl amino) -D-glucos described above.
  • the NMR, ELSD, and Mass results of 1-deoxy-1- (2-hydroxyethyl amino) -D-glusy prepared according to the preparation of the present invention are shown in FIGS.
  • the production method of the present invention was much superior to the conventional method using palladium in the yield (Table 1).
  • the manufacturing method of the present invention does not undergo a hydrogenation reaction process that is dangerously explosive even in the manufacturing process, and industrially yields 1-deoxy-1- (2-hydroxyethylamino) -D-glucos with high yield. Has the advantage to produce.
  • Example 2 Production of 6-Hydroxyethyl Amino-L-Libos by Bioconversion Reaction
  • 1-deoxy-1- (2-hydroxyethyl amino) -D prepared in Example 1, comprising 5 g / of magnesium sulfate, 5 g / of potassium monophosphate, and 5 g / of potassium diphosphate as a conversion medium composition.
  • a GSYN medium containing 10 g / i, 50 g / I, 100 gl i, 150 gl l and 200 g /, respectively, and dispense 50 in each of 500 Erlenmeyer flasks at 121 ° C. Sterilized for 30 minutes at.
  • the filtrate was adsorbed onto a strong acid cationic resin (Dowex 50X2-200), eluted with methanol and 30% aqueous ammonia solution and concentrated.
  • the concentrate was crystallized and recrystallized using methanol, dried and analyzed by gas chromatography to obtain 99.6% pure Migly from 62% yield from amino-L-sorbose to be 6-hydroxyl. It was confirmed.

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Abstract

The present invention relates to a method for preparing 1-deoxy-1-(2-hydroxyethyl amino)-D-glucitol from D-glucitol by using a borohydride complex as a reducing agent, and to a method for preparing miglitol from the 1-deoxy-1-(2-hydroxyethyl amino)-D-glucitol. The present invention has valuable use in the preparation of miglitol which is a therapeutic agent for diabetes.

Description

【명세서】  【Specification】
【발명의 명칭】  [Name of invention]
1-데옥시 -1— (2-하이드록시에틸 아미노) -D-글루시를 및 미글리를의 제조방법  Method for preparing 1-deoxy-1— (2-hydroxyethyl amino) -D-gluches and miglyls
【가술 분야】 [Artical field]
본 발명은 수소화붕소착물을 환원제로 사용하여 으글루시를로부터 1- 데옥시 -1— (2-하이드톡시에틸 아미노)ᅳ D—글루시를을 제조하는 방법 및 상기 1-데옥시 -1-(2-하이드록시에틸 아미노) -D-글루시를로부터 미글리를을 제조하는 방법에 관한 것이다.  The present invention provides a process for preparing 1-deoxy-1— (2-hydroxyethylamino) ᅳ D—glucile from agglutination using a borohydride complex as a reducing agent and the 1-deoxy-1- It relates to a process for producing migli from (2-hydroxyethyl amino) -D-glucos.
【배경 기술】 [Background technology]
미글리를은 바이엘 (Bayer)사에 의해 개발된 α_글루코시다제 ( α- glucosidase) 억제제로 1996년 FDA 승인을 얻었으며， 게 2형 당뇨병 치료를 위해 글리셋 (Glyset) 이라는 상품명으로 현재 판매되고 있다.  Miglyul is an α-glucosidase inhibitor developed by Bayer and obtained FDA approval in 1996 and is currently sold under the trade name Glyset for the treatment of crab type 2 diabetes. It is becoming.
산업적으로 미글리틀의 제조공정은 [반웅식 2]와 같이 포도당과 에탄을아민으로부터 1-데옥시 -1-(2-하이드록시에틸 아미노 )-D-글루시를을 합성하는 첫 번째 단계， 미생물을 생촉매로 사용하여 6-하이드록시에틸 아미노 -L-소르보스로 생물전환을 수행하는 두 번째 단계 및 화학합성을 통하여 미글리를을 합성하는 세 번째 단계를 거쳐 합성된다 ra/Jcei/ Materials Research, 197-198： p.51-55(2011) ) . Industrially, the manufacturing process of Miglet is the first step in synthesizing 1-deoxy-1- (2-hydroxyethyl amino) -D-glucose from glucose and ethane from amines, as shown in [Bannung 2]. Is synthesized through the second step of bioconversion to 6-hydroxyethyl amino-L-sorbose using the biocatalyst and the third step of synthesizing Migly through chemical synthesis ra / Jcei / Materials Research , 197-198: p. 51-55 (2011)).
시를 Poems
Figure imgf000004_0001
Figure imgf000004_0001
-0(-L-
Figure imgf000004_0002
미글리를 제조에 관한 -0 ( -L-
Figure imgf000004_0002
Regarding manufacturing Migley
짠행^헙 Tetrahedron Letters, 52： pp.3802- Tetrahedron Letters, 52 ： pp.3802-
3804( 2011))에 개시된 전합성 공정은 [반웅식 3]과 같다. 3804 (2011)), the presynthesis process is the same as [banung 3].
[반웅식 3]  [Banungsik 3]
Figure imgf000004_0003
위 문헌에 따르면， 전합성 공정은 분자 내 키랄 중심 (chiral center)에 대한 화학합성의 비선택성으로 인하여 보호기와 다단계의 복잡한 반웅이 필요하여 고가의 시약을 다량 사용하기 때문에 경제성이 거의 없다. 이 점올 고려하면， 미글리를 제조 시 위에서 명시된 1-데옥시— 1-(2- 하이드록시에틸 아미노) -D-글루시를로부터 6-하이드록시에틸 아미노 -L- 소르보스로의: 생물전환 반응이 중요한 공정임을 알 수 밌고, 이 생물전환 반웅에 필요한 중간체인 1-데옥시— 1-(2-하이드록시에틸 아미노) -D- 글루시틀을 제조하는 방법으로는 [반응식 4]와 같은 팔라듐-온- 차콜 (palladium-on-charcoal)올 사용하는 수소화 반웅이 있다 (EP0447160) .
Figure imgf000004_0003
According to the above literature, the presynthesis process requires little protection and multi-step complex reactions due to the nonselectivity of chemical synthesis for the chiral center in the molecule, so that the use of a large amount of expensive reagents is almost economical. Considering this oligo, Migly is prepared from 1-deoxy— 1- (2-hydroxyethyl amino) -D-glusy to 6-hydroxyethyl amino-L-sorbose specified above: It can be seen that the reaction is an important process, and the method for preparing 1-deoxy— 1- (2-hydroxyethyl amino) -D-glucit, an intermediate required for this bioconversion reaction, is shown in [Scheme 4]. There is a hydrogenation reaction using palladium-on-charcoal (EP0447160).
[반웅식 4]  [Banungsik 4]
Figure imgf000005_0001
Figure imgf000005_0001
[반응식 4]와 같은 방법에 따르면， 1-데옥시 -1-(2ᅳ하이드록시에틸 아미노) -D-글루시틀을 제조하기 위하여 12N HC1과 수소화 반웅을 위한 촉매로 팔라듐—온-차콜을 사용하였고， 온도 60±5°C와 4기압의 조건에서 반웅을 진행한다. 그러나， 수소화 반응은 폭발 위험성이 존재하기 때문에 산업적으로 적용하기 어렵다. 또한， 산성 조건에서 고온으로 인한 당의 갈변화로 인하여 결정화가 어렵고 수율이 비교적 낮은 문제가 있다. 본 명세서 전체에 걸쳐 다수의 논문 및 특허문헌이 참조되고 그 인용이 표시되어 있다. 인용된 논문 및 특허문헌의 개시 내용은 그 전체로서 본 명세서에 참조로 삽입되어 본 발명이 속하는 기술 분야의 수준 및 본 발명의 내용이 보다 명확하게 설명된다. 【발명의 내용】 According to the same method as in [Scheme 4], palladium-on-charcoal was used as a catalyst for hydrogenation reaction with 12N HC1 to prepare 1-deoxy-1- (2 ᅳ hydroxyethyl amino) -D-glucityl. The reaction was performed at a temperature of 60 ± 5 ° C and 4 atmospheres. However, hydrogenation reactions are difficult to apply industrially because of the risk of explosion. In addition, crystallization is difficult and yield is relatively low due to the brown change of sugar due to high temperature in acidic conditions. Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained. [Content of invention]
【해결하려는 과제】  [Problem to solve]
본 발명자들은 미글리를 제조에 필요한 중간체인 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D-글루시를을 고수율로 생산할 수 있^며， 상기한 바와 같은 종래 제조방법의 생산공정상의 위험성 및 저수율의: 문제점을 개선할 수 았는 신규한 제조방법을 개발하기 위하여 노력하였다. 그 결과, 에탄올아민을 포함하는 유기용매 하에서 수소화붕소착물을 환원제로 사용하는 경우， 반웅의 폭발 위험성 없이 고수율로 미글리를 중간체를 생산할 수 있음을 확인함으로써 , 본 발명을 완성하였다. The present inventors can produce in high yield 1-deoxy-1- (2-hydroxyethyl amino) -D-glusy, which is an intermediate for preparing Migly. Efforts have been made to develop new manufacturing methods which have been able to ameliorate the problems of the production process risks and low yields. As a result, when the borohydride complex is used as a reducing agent in an organic solvent containing ethanolamine, the present invention was completed by confirming that the intermediates can be produced in high yield without the risk of explosion.
따라서， 본 발명의 목적은 1-데옥시 -1-(2-하이드록시에틸 아미노) -D- 글루시를의 제조방법을 제공하는 데 있다.  It is therefore an object of the present invention to provide a process for the preparation of 1-deoxy-1- (2-hydroxyethyl amino) -D-gluches.
본 발명의 다른 목적은 미글리를의 제조방법을 제공하는 데 있다. 본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명， 청구범위 및 도면에 의해 보다 명확하게 된다.  Another object of the present invention is to provide a method for producing Migly. Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.
【과제의 해결 수단】 [Measures of problem]
본 발명의 일 양태에 따르면， 본 발명은 에탄을아민을 포함하는 유기용매 하에서， 수소화붕소착물을 환원제로 사용하여 D-글루시를을 하기 화학식 1의 1-데옥시 -1-(2-하이드록시에틸 아미노 )-D-글루시를로 제조하는 것을 특징으로 하는， 1-데옥시 -1-(2ᅳ하이드록시에틸 아미노) -D-글루시를의 제조방법을 제공한다:  According to an aspect of the present invention, the present invention is a ethanol under an organic solvent containing an amine, using the borohydride complex as a reducing agent to the D-glucin 1-deoxy -1- (2-hydride of formula 1 Provided is a process for preparing 1-deoxy-1- (2'hydroxyethyl amino) -D-glucy, characterized in that oxyethyl amino) -D-glucos is prepared by:
[화학식 1]  [Formula 1]
Figure imgf000006_0001
본 발명자들은 미글리를 제조에 필요한 중간체인 1-데옥시 -1ᅳ(2- 하이드록시에틸 아미노) -D-글루시를을 고수율로 생산할 수 있으며， 상기한 바와 같은 종래 제조방법의 생산공정상의 위험성 및 저수율의 문제점을 개선할 수 있는 신규한 제조방법을 개발하기 위하여 노력하였다. 그 결과， 에탄을아민을 포함하는 유기용매 하에서 수소화붕소착물을 환원제로 사용하는 경우, 반응의 폭발 위험성 없이 고수율로 미글리를 중간체를 생산할 수 있음을 확인였다. 본 발명의 가장 큰 특징은 1-데옥시— 1-(2-하이드록시에틸 아미노 )-D- 글루시를을 제조하는 과정에 있어 폭발 위험성이 있는 수소화 반웅을 위한 팔라듐ᅳ온-차콜을 사용하는 대신에 수소화붕소착물을 환원제로서 이용한다는 점이다. 본 발명의 제조방법은 하기 반웅식 1에 나타내었다.
Figure imgf000006_0001
The inventors of the present invention can produce 1-deoxy-1 '(2-hydroxyethyl amino) -D-glusy, which is an intermediate for preparing Migly, in high yield, and the production process of the conventional manufacturing method as described above. Efforts have been made to develop new methods of manufacturing that could improve the risks of phase and problems of low yield. As a result, it was confirmed that when the borohydride complex was used as a reducing agent in an organic solvent containing ethane, amines could be produced in high yield without a risk of explosion of the reaction. The biggest feature of the present invention is the use of palladium-ion-charcoal for the hydrogenation reaction with the risk of explosion in the preparation of 1-deoxy— 1- (2-hydroxyethyl amino) -D-glue. Instead, the borohydride complex is used as a reducing agent. The preparation method of the present invention is shown in the following reaction formula 1.
[반응식 1]  Scheme 1
Figure imgf000007_0001
Figure imgf000007_0001
D-글루코스 1-데옥시 -1-(2-하이드록시에릴아미노) -D-글루시를  D-glucose 1-deoxy-1- (2-hydroxyerylamino) -D-glucos
하기 실시예에서 증명된 바와 같이， 수소화붕소착물을 이용하는 본 발명의 제조방법은 종래의 팔라듐 -온-차콜을 이용하는 제조방법에 비하여 1-데옥시 -1-(2ᅳ하이드록시에틸 아미노) -D-글루시를의 생산수율을 현저히 상승시킨다 (표 1 참조). As demonstrated in the following examples, the preparation method of the present invention using the borohydride complex is 1-deoxy-1- (2 ᅳ hydroxyethyl amino) -D compared to the preparation method using the conventional palladium-on-charcoal. -Significantly increase the production yield of glue (see Table 1).
본 발명의 일구현예에 따르면, 본 발명의 제조방법에 따른 1-데옥시ᅳ 1-(2-하이드록시에틸 아미노) -으글루시를의 생산수율은 70- According to one embodiment of the invention, the production yield of 1-deoxysulf 1- (2-hydroxyethyl amino) -euglucin according to the preparation method of the present invention is 70-
10 。(이론수율)이며， 하나의 특정예에서는 70-90%이고 다른 특정예에서는 70-85%이며， 또 다른 특정예에서는 7으82¾이고; 또 다른 특정예에서는 75ᅳ 100%이고， 또 다른 특정예에서는 75-90%이며, 또 다른 특정예에서는 75- 85%이며， 또 다른 특정예에서는 75— 82%이며; 또 다른 특정예에서는 8으 100%이고ᅳ 또 다른 특정예에서는 80-90%이며, 또 다른 특정예에서는 8으 85%이고， 또 다른 특정예에서는 80-82%이다. 10. (theoretical yield), 70-90% in one specific example, 70-85% in another specific example, 782¾ in another specific example; In another specific example 75% 100%, in another specific example 75-90%, in another specific example 75-85%, in another specific example 75-82%; In another specific example 8 is 100% and in another specific example 80-90%, in another specific example 8 is 85% and in another specific example 80-82%.
본 발명의 일구현예에 따르면， 상기 ' 수소화붕소착물은 수소화붕소나트륨 (NaB¾), 수소화붕소시아노나트륨 (NaCNB¾) 및 디메틸아미노보란 (BH4N(CH3)2)으로 구성된 군으로부터 선택된다. According to one embodiment of the present invention, the ' borohydride complex is selected from the group consisting of sodium borohydride (NaB¾), sodium borohydride (NaCNB¾) and dimethylaminoborane ( BH 4 N ( C H 3) 2) .
하나의 특정예에서， 상기 수소화붕소착물은 수소화붐소나트륨 또는 수소화붕소시아노나트륨이고， 다른 특정예에서는 수소화붕소나트륨이다. 본 발명의 제조방법에서， 수소화붕소착물이 사용되는 양은 D- 글루코스의 몰수 대비 1 내지 10 당량이며， 하나의 특정예에서는 2 내지 5 당량이고， 다른 특정예에서는 2 내지 3 당량이다. . 본 발명의 일구현예에 따르면， 상기 유기용매는 암모늄염을 포함한다. 하나의 특정예에서, 상기 암모늄염은 아세트산 암모늄 (C¾C00NH4), 제 1인산 암모늄 (NH4H2P04), 제 2인산 암모늄 ((NH4)2HP04), 황산암모늄 ((NH4)2S04), 질산암모늄 (NH4N03) 및 탄산암모늄 ((NH4)2C03)으로 구성된 군으로부터 선택되며, 다른 특정예에서는 아세트산 암모늄이다. In one specific example, the borohydride complex is sodium borohydride or sodium borocyanohydride, and in another specific example, sodium borohydride. In the production method of the present invention, the amount of boron hydride used is 1 to 10 equivalents relative to the number of moles of D-glucose, in one particular example 2 to 5 equivalents, and in another specific example 2 to 3 equivalents. . According to one embodiment of the present invention, the organic solvent includes an ammonium salt. In one specific example, the ammonium salt is ammonium acetate (C¾C00NH 4 ), ammonium monophosphate (NH 4 H 2 P0 4 ), ammonium diphosphate ((NH 4 ) 2 HP0 4 ), ammonium sulfate ((NH 4 ) 2 S0 4 ), ammonium nitrate (NH 4 N0 3 ) and ammonium carbonate ((NH 4 ) 2 C0 3 ), in another specific example ammonium acetate.
■ 본 발명의 제조방법에서, 암모늄염이 사용되는 양은 D-글루코스와 몰수 대비 1 내지 20 당량이며， 하나의 특정예에서는 3 내지 15 당량이고， 다른 특정예에서는 5 내지 10 당량이다. In the production method of the present invention, the amount of ammonium salt is used is 1 to 20 equivalents based on D-glucose and the number of moles, 3 to 15 equivalents in one specific example, and 5 to 10 equivalents in another specific example.
본 발명의 일구현예에 따르면， 상기 유기용매는 메틸알코올， 에틸알코올, 프로필 알코을， 이소프로필 알코을과 같은 알코을계 용매; 에틸 아세테이트를 포함하는 에스테르계 용매; 메틸렌 클로라이드 및 클로로포름을 포함하는 할로 알칸계 용매; 테트라히드로퓨란을 포함하는 에테르 용매; 아세톤 및 메틸에틸케톤을 포함하는 케론계 용매; 아세토니트릴: 및 프로피오니트릴을 포함하는 니트릴계 용매; 다메틸포름아미드 및 디메틸아세트아미드를 포함하는 아미드계 용매 ; 디메틸술폭시드를 포함하는 술폭시드계 용매; 상기 둘 이상의 유기용매가 흔합된 용매; 또는 상기 유기용매와 물이 흔합된 용매이다.  According to one embodiment of the invention, the organic solvent is an alcohol solvent such as methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol; Ester solvents including ethyl acetate; Halo alkanes solvents including methylene chloride and chloroform; Ether solvents including tetrahydrofuran; Keron solvents including acetone and methyl ethyl ketone; Acetonitrile: nitrile solvents including propionitrile; Amide solvents including dimethylformamide and dimethylacetamide; Sulfoxide solvents including dimethyl sulfoxide; A solvent in which the two or more organic solvents are mixed; Or a solvent in which the organic solvent and water are mixed.
하나의 특정예에서， 상기 유기용매는 알코을계 용매이고, 다른 특정예에서 상기알코을계 용매는 에틸알코을이다.  In one specific example, the organic solvent is an alcoholic solvent, and in another specific example, the alcoholic solvent is ethyl alcohol.
본 발명의 제조방법에서 사용되는 용매의 부피는 D-글루코스의 질량 (g) 대비 부피 (m«비로서 1:1-20이며, 하나의 특정예에서는 1:1-15이고， 다른 특정예에서는 1:5-10이다.  The volume of the solvent used in the preparation method of the present invention is 1: 1-20 as the volume (m «ratio) relative to the mass (g) of D-glucose, in one specific example 1: 1-15, and in another specific example 1: 5-10.
본 발명의 일구현예에 따르면, 본 발명의 제조방법은 팔라듐ᅳ온- 차콜을 이용하는 종래의 제조방법에 비하여 낮은 온도에서 반응을 진행한다. 하나의 특정예에서， 본 발명의 반웅공정은 -20 내지 60°C의 온도에서 진행하며， 다른 특정예에서는 0 내지 4(rc의 온도에서 진행하고, 또 다른 특정예에서는 20 내지 40°C의 온도에서 진행한다. According to one embodiment of the present invention, the production method of the present invention proceeds the reaction at a lower temperature than the conventional production method using palladium ᅳ -charcoal. In one specific example, the reaction process of the present invention proceeds at a temperature of -20 to 60 ° C, in another specific example 0 to 4 (running at a temperature of rc, and in another specific example of 20 to 40 ° C) Proceed at temperature.
본 발명의 제조방법에서， 반응시간은 5분 내지 24시간이고， 하나의 특정예에서는 30분 내지 5시간이며， 다른 특정예에서는 30분 내지 1시간이다.  In the production method of the present invention, the reaction time is 5 minutes to 24 hours, in one specific example is 30 minutes to 5 hours, and in another specific example is 30 minutes to 1 hour.
본 발명의 제조방법에서는 1-데옥시 -1-(2-하이드록시에틸 아미노) - D-글루시를의 단리 및 정제를 위하여， 이온교환제， 실리카겔 또는 실릴화 실리카겔로 크로마토그래피시켜서 각 경우의 생성물 함유용액을 농축하고， 이를 적당한 용매로 결정화 할 수 있다. In the preparation method of the present invention, 1-deoxy-1- (2-hydroxyethyl amino)- For the isolation and purification of D-gluches, the product-containing solution in each case can be concentrated by chromatography with an ion exchanger, silica gel or silylated silica gel and crystallized with a suitable solvent.
본 발명의 적합한 이온교환제는 약산성， 강산성, 약염기성 및 강염기성 형태 모두를 포함하며, 하나의 특정예에서는 강산성 또는 강염기성 이온교환제이다.  Suitable ion exchangers of the present invention include both weakly acidic, strongly acidic, weakly basic and strongly basic forms, and in one particular example are strongly acidic or strongly basic ion exchangers.
본 발명의 적합한 결정화용매는 에틸 아세테이트와 같은 에스테르계 용매; 메틸렌 클로라이드, 클로로포름과 같은 할로 알¾계 용매; 테트라히드로퓨란과 같은 에테르 용매; 아세톤， 메틸에틸케톤과 같은 케톤계 용매; 아세토니트릴, 프로피오니트릴과 같은 니트릴계 용매; 메틸알코올, 에틸알코올， 프로판올， 이소프로판을과 같은 알코을계 용매; 및 이들의 둘 이상이 흔합된 유기 용매들을 포함한다.  Suitable crystallization solvents of the present invention include ester solvents such as ethyl acetate; Halo al¾ solvents such as methylene chloride and chloroform; Ether solvents such as tetrahydrofuran; Ketone solvents such as acetone and methyl ethyl ketone; Nitrile solvents such as acetonitrile and propionitrile; Alcohol solvents such as methyl alcohol, ethyl alcohol, propanol and isopropane; And organic solvents in which two or more thereof are mixed.
하나의 특정예에서， 상기 결정화 용매는 에틸알코을이다. 결정화 용매의 사용되는 부피는 사용되는 D-글루코스의 중량 (g) 대비 부피 ( )비로써 1.0 내지 20배이며， 하나의 특정예에서는 5 내지 10배이다. 이러한 반웅공정의 진행 정도는 박막 크로마토그래피 또는 ELSD (Evaporative Light Scattering Detector)등의 수단을 사용하여 분석 및 측정할 수 있다. 본 발명의 다른 양태에 따르면， 본 발명은 다음의 단계를 포함하는 미글리를 (Miglitol)의 제조방법을 제공한다:  In one specific example, the crystallization solvent is ethyl alcohol. The volume used of the crystallization solvent is from 1.0 to 20 times by volume () ratio to the weight (g) of D-glucose used, in one particular example from 5 to 10 times. The progress of the reaction process can be analyzed and measured using a means such as thin layer chromatography or ELSD (Evaporative Light Scattering Detector). According to another aspect of the present invention, the present invention provides a method for producing Miglitol, comprising the following steps:
(a) 에탄을아민을 포함하는 유기용매 하에서， 수소화붕소착물을 환원제로 사용하여 D-글루시를을 상기 화학식 1의 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D-글루시를로 제조하는 단계;  (a) Under the organic solvent containing ethane, amine, using the borohydride complex as a reducing agent to the D-glucin to 1-deoxy-1- (2-hydroxyethyl amino) -D-glu Preparing the pore;
(b) 단계 (a)에서 제조한 1-데옥시 -1-(2-하이드록시에틸 아미노) -D- 글루시를을 글루코노박터 속 균주 또는 이의 휴면세포 (resting cell)를 이용하여 6-하이드록시에틸 아미노 -L-소르보스로 생물전환하는 단계; 및  (b) 1-deoxy-1- (2-hydroxyethyl amino) -D-glucos prepared in step (a) using 6-glucobacterium strain or resting cells thereof. Bioconversion to hydroxyethyl amino-L-sorbose; And
(c) 단계 (b)에서 생물전환 된 6-하이드톡시에틸 아미노 -L- 소르보스를 미글리를로 합성하는 단계 .  (c) synthesizing 6-hydroxyethyl amino-L-sorbose biomigrated in step (b) into Migly.
본 발명의 미글리를의 제조방법은 상술한 1-데옥시 -1-(2- 하이드록시에틸 아미노 )-D-글루시를의 제조방법을 이용하기 때문에， 이 둘 사이에 공통된 내용은 본 명세서의 과도한 복잡성을 피하기 위하여 그 기재를 생략한다. Since the method for producing Miglyol of the present invention uses the above-described method for preparing 1-deoxy-1- (2-hydroxyethyl amino) -D-glycine, these two The common content between them is omitted in order to avoid excessive complexity of the present specification.
본 발명의 일구현예에 따르면， 단계 (b)는 글루코노박터 속 균주 또는 상기 미생물의 휴면세포를 1-데옥시 -1-(2-하이드록시에틸 아미노) -D- 글루시를을 함유하는 배지에서 배양하여 실시한다. 이때， 1-데옥시 -1-(2- 하아드톡시에릴 아미노) -D-글루시를은 균주를 배지에 접종하기에 앞서 첨가하거나 균주의 접종 후에 첨가할 수 있으며， 배양 시작과 동시에 첨가하거나 또는 배양 과정에 걸쳐 연속적으로 또는 분할해서 첨가할 수 있다. 사용되는 1-데옥시 -1-(2-하이드록시에틸 아미노) -으글루시를의 양은 당업자에 의해 실험적으로 결정될 수 있다.  According to one embodiment of the present invention, step (b) comprises a 1-deoxy-1- (2-hydroxyethyl amino) -D-glucose of dormant cells of the bacterium Gluconobacter or the microorganism. Incubate in medium. In this case, 1-deoxy-1- (2-hadoxyethylamino) -D-glucy may be added before inoculating the silver strain or after inoculation of the strain, and added at the same time as the start of the culture. Or it can be added continuously or in portions throughout the culture process. The amount of 1-deoxy-1- (2-hydroxyethyl amino) -agglucius used can be determined experimentally by one skilled in the art.
하나의 특정예에서， 6_하이드록시에틸 아미노 -L-소르보스의 제조를 위한 배지는 1-데옥시ᅳ1-(2-하이드록시에틸 아미노) -D-글루시를을 10 gl 내지 150 의 농도로 함유할 수 있고, 다른 특정예에서는 10 gl i 내자 100 g/£의 농도로 함유할 수 있으며， 또 다른 특정예에서는 10 gl i 내지 80 g/ 의 농도로 함유할 수 있다. In one particular example, _ 6-hydroxy-ethylamino -L- medium for the production of a sorbose is a 1-deoxy-eu 1- (2-hydroxy-ethylamino) -D- gluconic when the the 10 to 150 gl It may be contained in a concentration, in another specific example may be contained in a concentration of 100 g / 내 within 10 gl i, in another specific example may be contained in a concentration of 10 gl i to 80 g /.
균주를 휴면세포 페이스트 (resting cell paste)의 형태로 사용할 경우의 상기 배지는 1-데옥시— 1ᅳ(2-하이드록시에틸 아미노) -D-글루시를을 10 g/H 내지 200 농도로 함유할 수 있고, 다른 특정예에서는 10 g/ i 내지 150 의 농도로 함유할 수 있다.  When the strain is used in the form of a resting cell paste, the medium contains 1-deoxy—1 ′ (2-hydroxyethyl amino) -D-glusy at a concentration of 10 g / H to 200. In another specific example, it may contain at a concentration of 10 g / i to 150.
본 발명에 제조방법에 사용되는 배지는 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D-글루시를뿐만 아니라, D-글루시를, 질소원 무기염 및 미량원소， 성장인자를 포함할 수 있으며， 적어도 하나의 다른 탄소원을 포함할 수도 있다. 첨가되는 각 성분의 양은 6-하이드록시에틸 아미노 -L-소르보스의 생성올 최대화할 수 있도록 선택되는 것이 바람직하며， 그 양은 당해 기술분야에 알려진 여러 가지 방법에 따라 당업자에 의해 실험적으로 결정될 수 있다. 1-데옥시 -1-(2-하이드톡시에틸 아미노) 글루시를로부터 6-하이드록시에틸 아미노 -L-소르보스로의 생산을 극대화하기 위한 배양조건으로서 온도, pH, 통기， 교반， 배양기간 등의 최적화는 당업자에 의해 실험적으로 결정될 수 있다. 특정 배양조건의 선택은 사용되는 균주， 휴면세포， 배지조성, 배양규모 및 기타 고려사항 등에 따라 달라질 수 있다. 본 발명의 일구현예에 따르면， 본 발명의 제조방법에서 균주의 배양온도는 15 내지 37°C에서 이루어지는데， 하나의 특정예에서는 25°C에서 이루어진다. 배양액의 적절한 pH는 4.0 내지 8.0 범위이며， 하나의 특정예에서는 4.5 내지 7.0 범위이고, 다른 특정예에서는 5.5 내지 6.0이다. 본 발명에서는， 6-하이드록시에틸 아미노 -L-소르보스의 제조를 위하여 글루코노박터 속 균주의 세포추출물 또는 상기 미생물의 휴면세포꾀 세포추출물을 이용할 수도 있다. 세포추출물올 이용한 6-하이두록시에틸 아미노 -L—소르보스의 제조방법은 (a) 글루코노박터 속 균주의 세포추출물 또는 상기 미생물의 휴면세포의 세포추출물을 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D-글루시를에 접촉시키는 단계; 및 (b) 6- 하이드록시에틸 아미노 -L-소르보스를 회수하는 단계를 포함하여 수행된다. 본 발명의 일구현예에 따르면， 상기 글루코노박터 속 균주는 글루코노박터 옥시단스이고, 하나의 특정예에서는 글루코노박터 옥시단스 CKDBM 0901아다. 상기 글루코노박터 옥시단스 CKDBM 0901은 1—데옥시 -1-(2- 하이드톡시에틸 아미노) -D-글루시를에 대한 내성 및 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D-글루시를을 6-하이드록시에틸 아미노 -L- 소르보스로 고수율로 생물전환하는 능력을 갖는 균주로서， 2012년 9월 21일자로 한국생명공학연구원 생물자원센터 (Korean Collection for Type Cultures, KCTC)에 기탁번호 KCTC 12282BP로 기탁되었다. 상기 글루코노박터 옥시단스 CKDBM 0901은 한국특허출원 제 10-2012-0112395호에 상세히 개시되어 있으며， 상기 특허문헌은 본 명세서에 참조로서 ：삽입된다. 본 발명의 단계 (c)에서는 생물전환 된 6-하이드록시에틸 아미노 -L- 소르보스를 미글리틀로 합성한다. 상기 미글리를은 (2R,3R,4R,5S)-l-(2- hydroxy ethyl )-2-(hydroxymethyl )piper i dine— 3,4,5— trk)l로서 α― 글루코시다제 억제제이며, 경구용 제 2형 당뇨병 치료제이다. The medium used in the preparation method of the present invention is not only 1-deoxy-1- (2-hydroxyethyl amino) -D-glue, but also D-glue, nitrogen source inorganic salts and trace elements, and growth factors. It may include, and may include at least one other carbon source. The amount of each component added is preferably selected to maximize the production of 6-hydroxyethyl amino-L-sorbose, and the amount can be determined experimentally by one skilled in the art according to various methods known in the art. . Culture conditions for maximizing the production of 1-deoxy-1- (2-hydroxyethyl amino) glue from 6-hydroxyethyl amino-L-sorbose as temperature, pH, aeration, stirring, incubation period Optimization can be determined experimentally by one skilled in the art. The choice of specific culture conditions may vary depending on the strain used, dormant cells, medium composition, culture size and other considerations. According to one embodiment of the invention, the culture temperature of the strain in the production method of the present invention is made at 15 to 37 ° C, in one specific example is made at 25 ° C. Suitable pH of the culture is in the range from 4.0 to 8.0, in one particular range from 4.5 to 7.0, in another particular range from 5.5 to 6.0. In the present invention, 6-hydroxyethyl amino-L-sorbose cell extract of the genus Gluconobacter strain or dormant cell extract of the microorganism may be used. The method for preparing 6-hyduoxyethyl amino-L-sorbose using cell extracts is (a) cell extracts of the genus Gluconobacter or cell extracts of dormant cells of the microorganism. Contacting hydroxyethyl amino) -D-gluci to; And (b) recovering 6-hydroxyethyl amino-L-sorbose. According to one embodiment of the present invention, the strains of the genus Gluconobacter are gluconobacter oxydans, and in one particular example, gluconobacter oxydans CKDBM 0901. The Gluconobacter Oxydans CKDBM 0901 is resistant to 1—deoxy-1- (2-hydroxyethylamino) -D-glucy and 1-deoxy-1- (2-hydroxyethyl amino)- A strain that has the ability to bioconvert D-glucis to 6-hydroxyethyl amino-L-sorbose in high yield, dated Sept. 21, 2012, Korean Collection for Type Cultures , KCTC) was deposited with the accession number KCTC 12282BP. The Gluconobacter oxydans CKDBM 0901 is disclosed in detail in Korean Patent Application No. 10-2012-0112395, which is incorporated herein by reference. In step (c) of the present invention, bio-converted 6-hydroxyethyl amino-L-sorbose is synthesized in Miglit. Miglyul is (2R, 3R, 4R, 5S) -l- (2-hydroxyethyl) -2- (hydroxymethyl) piper i dine— 3,4,5—trk) l, an α-glucosidase inhibitor Oral type 2 diabetes treatment.
6-하이드록시에틸 아미노 -L-소르보스를 사용하여 미글리를을 제조하는 방법은 미국특허 제 5 ,602， 013호， 미국특허 제 5 ,610 ,039호， 미국특허 제 5 ,916 ,748호 및 미국특허 제 6， 552， 176호에 기술되어 있으며, 상기 특허문헌은 참조로서 본 명세서에 삽입된다.  Methods for preparing migli using 6-hydroxyethyl amino-L-sorbose are described in U.S. Patent Nos. 5,602, 013, U.S. Patent 5,610,039, U.S. Patent 5,916,748 And US Patent Nos. 6,552, 176, which are incorporated herein by reference.
본 발명의 일구현예에 따르면, 단계 (b)에서 얻은 6-하이드록시에틸 아미노 -L-소르보스를 이용하여 미글리를을 합성하는 방법은 6- 하이드록시에틸 아미노 -L-소르보스를 포함하는 배양액을 여과한 다음 활성탄을 처리하여 색소를 제거하고 수소화 반웅기로 이송하여: 미글리를 합성을 수행하여 실시된다. 수소화 반웅을 위한 촉매로는 팔라듐「온-차콜을 사용할 수 있고, 반웅에 적합한 온도와 압력조건에서 반응을 진행한 다음 여과하여 촉매를 제거한다. 상기 여과액으로부터 미글리롤을 정제하기 위하여 수지에 흡착 후 용리하고 농축한 다음 결정화와 재결정화 과정을 거쳐 건조하면， 순도가 높은 미글리를올 백색의 분말형태로 얻을 ^ 있다. According to one embodiment of the present invention, the method for synthesizing migli using 6-hydroxyethyl amino-L-sorbose obtained in step (b) is 6- The culture solution containing hydroxyethyl amino-L-sorbose was filtered and then treated with activated carbon to remove the pigment and transferred to a hydrogenation reactor: Migley was carried out by carrying out the synthesis. Palladium "on-charcoal may be used as a catalyst for hydrogenation reaction, and reaction is performed at the temperature and pressure conditions suitable for reaction, and it removes a catalyst by filtering. In order to purify miglilol from the filtrate, the mixture was eluted with a resin, eluted and concentrated, and then dried through a crystallization and recrystallization process to obtain a high purity migliol in the form of an all-white powder.
【발명의 효과】 【Effects of the Invention】
본 발명의 특징 및 이점을 요약하면 다음과 같다:  The features and advantages of the present invention are summarized as follows:
(i) 본 발명은 수소화붕소착물을 환원제로 사용하여 D- 글루시를로부터 1-데옥시ᅳ1-(2-하이드톡시에틸 아미노) -D-글루시를을 제조하는 방법, 및 제조된 1-데옥시-； L-(2-하이드톡시에틸 아미노) -D- 글루시틀을 생물전환하여 미글리를을 제조하는 방법을 제공한다. (i) The present invention provides a process for preparing 1-deoxy ᅳ 1- (2-hydroxyethylamino) -D-glue from D-glucos using boron hydride complex as reducing agent, and 1 prepared -Deoxy-; L- ( 2 -hydroxyethyl amino) -D-gluclet is bioconverted to provide a method for producing migli.
(ii) 본 발명의 제조방법은 팔라듐-온 -차콜 대신 수소화붕소착물을 사용함으로써 폭발 위험성이 있는 수소화 반웅공정올 거치지 않고도 높은 수율로 1-데옥시 -1-(2-하이드록시에틸 아미노 )-D—글루시를올 생산할 수 있다.  (ii) The preparation method of the present invention uses boron hydride complexes instead of palladium-on-charcoal to yield 1-deoxy-1- (2-hydroxyethyl amino)-in high yield without the risk of explosion. D—Can produce glue.
(iii) 따라서， 본 발명은 당뇨병 치료제인 미글리를의 제조를 위하여 유용하게 사용될 수 있다.  (iii) Therefore, the present invention can be usefully used for the production of Migly, a therapeutic agent for diabetes.
【도면의 간단한 설명】 [Brief Description of Drawings]
도 1은 본 발명의 방법에 따라 제조된 1-데옥시 -1-(2-하이드록시에틸 아미노) -D-글루시를에 .대한 NMR 결과이다.  Figure 1 is an NMR result for 1-deoxy-1- (2-hydroxyethyl amino) -D-glucos prepared according to the method of the present invention.
도 2는 본 발명의 방법에 따라 제조된 1ᅳ데옥시 -1-(2-하이드록시에틸 아미노 )-D-글루시를에 대한 ELSD (Evaporative Light Scattering Detector) 결과이다.  FIG. 2 shows the results of an Evaporative Light Scattering Detector (ELSD) for lnddeoxy-1- (2-hydroxyethyl amino) -D-glucis prepared according to the method of the present invention.
도 3은 본 발명의 방법에 따라 제조된 1-데옥시 -1-(2-하이드록시에틸 아미노 )-D-글루시틀에 대한 Mass 결과이다.  FIG. 3 is a Mass result for 1-deoxy-1- (2-hydroxyethyl amino) -D-glusytle prepared according to the method of the present invention. FIG.
도 4는 본 발명의 방법에 따라 제조된 미글리를에 대하여 적외선 분광법으로 분석한 스펙트럼을 나타낸 것이다. 도 5는 본 발명의 방법에 따라 제조된 미글리를에 대한 NMR 결과이다. Figure 4 shows the spectrum analyzed by infrared spectroscopy for Migley prepared according to the method of the present invention. 5 is an NMR result for Migly prepared according to the method of the present invention.
【발명을 실시하기 위한 구체적인 내용】 [Specific contents to carry out invention]
이하， 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다. 실시예  Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. . Example
실시예 1. 1-데옥시 -1-(2-하이드록시에틸 아미노) -D-글루시를의 제조 수소화붕소나트륨을사용한제조 (제조예 Q)  Example 1. Preparation of 1-deoxy-1- (2-hydroxyethyl amino) -D-glucido Preparation using sodium borohydride (Preparation Example Q)
2 용량 플라스크에 아세트산 암모늄 214g을 녹인 에틸알코을 1,000 m«와 D-글루코스 100g을 넣고， 약 5°C로 넁각시켰다. 여기에 에탄을아민 35g을 넣고， 수소화붕소나트륨 (NaBH4) 63g을 5-KTC의 은도에서 천천히 가한 후， 상온으로 온도를 서서히 상승시켜 60분간 교반한 다음 반응을 종결하였다. 이 반응액에 5% 염산 50 를 넣어주고， 10분¾ 상온에서 교반한 다음 정치시켰다. 종결된 반응액은 NAC— 4 강산성 양이온 교환제에 흡착시킨 다음 정제수로 세척하고, 이어서 30% 암모니아수와 메틸알코을 용리액으로 용출하고 농축하였다. 생성 분획물을 진공 농축시켜 얻어진 잔사를 에틸알코올로부터 결정화하고 여과한 후 건조하여 1-데옥시 -1-(2ᅳ 하이드록시에틸 아미노) -D-글루시를 102g (이론수율 82%)을 얻었다 (순도: 99.9%). 1,000 m «and 100 g of D-glucose were added to ethyl alcohol, which was dissolved 214 g of ammonium acetate in a two-volume flask, and was cooled to about 5 ° C. 35 g of amine amine was added thereto, and 63 g of sodium borohydride (NaBH 4 ) was slowly added at a temperature of 5-KTC, and the temperature was gradually raised to room temperature, followed by stirring for 60 minutes, and then the reaction was terminated. 50% of 5% hydrochloric acid was added to the reaction solution, and the mixture was stirred for 10 minutes at room temperature and then left to stand. The terminated reaction solution was adsorbed onto NAC-4 strong acid cation exchanger and washed with purified water, followed by eluting 30% aqueous ammonia and methyl alcohol with eluent and concentrating. The resulting fractions were concentrated in vacuo and the residue obtained was crystallized from ethyl alcohol, filtered and dried to give 102 g (theoretical yield 82%) of 1-deoxy-1- (2 ′ hydroxyethyl amino) -D-glucy ( Purity: 99.9%).
¾ 丽 R (400 MHz, D20) δ 3.92-3.87 (m, 1H, H-3), 3.83-3.80 (dd, 1H, H-l'), 3.72-3.74 (m, 1H, H-2), 3.72-3.67 (m, 2H, H— 1' and H-4) , 3.65-3.61 (m, 2H, H-4 and H-6) , 2.82-2.64 (m, 4H, H-7, H_8, H-9, H- 10)； MS(ESI)m/z calcd for [C8Hi9N06] 225.12, obsd 225.12. 수소화붕소시아노나트륨을사용한제조 (제조예 ) ¾ δ R (400 MHz, D20) δ 3.92-3.87 (m, 1H, H-3), 3.83-3.80 (dd, 1H, H-1 '), 3.72-3.74 (m, 1H, H-2), 3.72-3.67 (m, 2H, H— 1 'and H-4), 3.65-3.61 (m, 2H, H-4 and H-6), 2.82-2.64 (m, 4H, H-7, H_8, H -9, H-10); MS (ESI) m / z calcd for [C 8 Hi 9 N0 6 ] 225.12, obsd 225.12. Preparation using sodium borohydride sodium (production example)
2ί 용량 플라스크에 아세트산 암모늄 (CH3C00NH4) 214g을 녹인 에틸알코올 1,000 ^와 D-글루코스 100g을 넣고, 약 5°C로 넁각시켰다. 여기에 에탄올아민 35g을 넣고， 수소화붕소 시아노나트륨 (NaCNBrf3) 103g을 5-KTC의 온도에서 천천히 가한 후， 상온으로 온도를 서서하 상승시켜 60분간 교반한 다음 반웅을 종결하였다. 이 반웅액에 5% 염산 50 ^를 넣어주고， 10분간 상온에서 교반한 다음 정치시켰다. 종결된 반웅액은 NAC- 4 강산성 양이온 교환제에 흡착시킨 다음 정제수로 세척하고， 이어서 30% 암모니아수와 메틸알코을 용리액으로 용출하고 농축하였다. 생성 분획물을 진공 농축시켜 얻어진 잔사를 에틸알코을로부터 결정화하고， 여과한 후 건조하여 1-데옥시 -1-(2-하이드록시에틸 아미노) -D-글루시를 100g (이론수을 80¾»)을 얻었다 (순도: 99.7%). 팔라듐을사용한제조 (비교예) 1,000 mL of ethyl alcohol and 100 g of D-glucose dissolved in 214 g of ammonium acetate (CH 3 C00NH 4 ) were added to a 2 ί flask, and the mixture was stirred at about 5 ° C. Add 35 g of ethanolamine, and 103 g of boron hydride cyano sodium (NaCNBrf 3 ). After slowly adding at a temperature of 5-KTC, the temperature was raised to room temperature and then lowered and stirred for 60 minutes, and then reaction was terminated. 50% of 5% hydrochloric acid was added to the reaction solution, the mixture was stirred at room temperature for 10 minutes, and then allowed to stand. The finished reaction solution was adsorbed on a NAC-4 strongly acidic cation exchanger, washed with purified water, and then eluted with 30% aqueous ammonia and methyl alcohol and concentrated. The resulting fractions were concentrated in vacuo and the residue obtained was crystallized from ethyl alcohol, filtered and dried to give 100 g of 1-deoxy-1- (2-hydroxyethyl amino) -D-glusy (80¾ »theoretical water). (Purity: 99.7%). Manufacture using palladium (comparative example)
21 수소화 반응기 (PARR Instrument Co. USA, 5100 Reactor)에 에틸알코올 1000 와 D-글루코스 100g을 넣은 후， 에탄올아민 35g과 4% 팔라듐-온 -차콜 (palladium-on-charcoal) 30g을 넣고， 수소압력을 6 bar로 조정한 후 60°C에서 24시간 교반하였다. 반웅이 종결된 후 여과를 실시하였다. 여과 후 에틸 알코을 50 를 사용하여 촉매를 세척하고， 여과된 반웅액은 진공 농축시켜 얻어진 잔사를 에틸알코올로부터 결정화하고， 여과한 후 건조하여 1-데옥시 -1-(2-하이드록시에틸 아미노 )-D- 글루시를 80g (이론수율 64%)을 얻었다 (순도: 99.5%). 비교결과 21 Pour 100 g of ethyl alcohol and 100 g of D-glucose into a hydrogenation reactor (PARR Instrument Co. USA, 5100 Reactor), add 35 g of ethanolamine and 30 g of 4% palladium-on-charcoal. After adjusting to 6 bar and stirred at 60 ° C. for 24 hours. Filtration was carried out after the reaction was complete. After filtration, the catalyst was washed with ethyl alcohol 50, and the filtrate reaction mixture was concentrated in vacuo to crystallize the residue obtained from ethyl alcohol, filtered and dried to obtain 1-deoxy-1- (2-hydroxyethyl amino). 80 g (64% yield) of -D-glue were obtained (purity: 99.5%). Comparison result
상기에서 설명한 1-데옥시 -1-(2-하이드록시에틸 아미노) -D- 글루시를의 제조예 ①， 제조예 ② 및 비교예에 따른 결과를 표 1에 나타내었다. 본 발명의 제조예에 의하여 제조된 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D-글루시를의 NMR, ELSD 및 Mass 결과는 도 1 내지 3에 나타내었다.  Table 1 shows the results of Preparation Example 1, Preparation Example 2, and Comparative Example of 1-deoxy-1- (2-hydroxyethyl amino) -D-glucos described above. The NMR, ELSD, and Mass results of 1-deoxy-1- (2-hydroxyethyl amino) -D-glusy prepared according to the preparation of the present invention are shown in FIGS.
【표 11  Table 11
Figure imgf000014_0001
표 1에 나타난 바와 같이， 본 발명의 제조방법은 수율에 있어 팔라듐을 이용한 종래의 방법에 비하여 훨씬 우수하몄다 (표 1). 또한， 본 발명의 제조방법은 제조공정상으로도 폭발 위험성이 있는 수소화 반응공정을 거치지 않고， 산업적으로 높은 수율로 1-데옥시 -1-(2- 하이드톡시에틸 아미노) -D-글루시를을 생산할 수 있는 이점을 갖는다. 실시예 2. 생물전환 반웅에 의한 6-하이드록시에틸 아미노 -L- ί르보스의 생산
Figure imgf000014_0001
As shown in Table 1, the production method of the present invention was much superior to the conventional method using palladium in the yield (Table 1). In addition, the manufacturing method of the present invention does not undergo a hydrogenation reaction process that is dangerously explosive even in the manufacturing process, and industrially yields 1-deoxy-1- (2-hydroxyethylamino) -D-glucos with high yield. Has the advantage to produce. Example 2 Production of 6-Hydroxyethyl Amino-L-Libos by Bioconversion Reaction
전환배지 조성으로 황산마그네슘 5 g/ , 제일인산칼륨 5 g/ 및 제이인산칼륨 5 g/ 를 포함하고, 실시예 1에서 제조한 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D—글루시를을 각각 10 g/i , 50 g/ I , 100 gl i， 150 gl l 및 200 g/ 로 포함하는 GSYN 배지를 제조한 다음 500 의 삼각 플라스크에 50 in씩 분주한 후 121°C에서 30분간 살균하였다. 상기와 같이 제조된 플라스크에 글루코노박터 옥시단스 CKDBM0901C기탁번 KCTC 12282BP)의 휴면세포 페이스트 (resting cell paste)를 1.5g씩 첨가하고， 1- 데옥시 -1-(2-하이드록시에틸 아미노) -D-글루시를 100 g/ 를 사용하여 25°C에서 220rpm으로 96시간 동안 전환배양을 실시하였다. 실시예 3. 6-하이드록시에틸 아미노 -L-소르보스로부터 미글리를의 합성 1-deoxy-1- (2-hydroxyethyl amino) -D prepared in Example 1, comprising 5 g / of magnesium sulfate, 5 g / of potassium monophosphate, and 5 g / of potassium diphosphate as a conversion medium composition. —Create a GSYN medium containing 10 g / i, 50 g / I, 100 gl i, 150 gl l and 200 g /, respectively, and dispense 50 in each of 500 Erlenmeyer flasks at 121 ° C. Sterilized for 30 minutes at. To the flask prepared as described above, add 1.5 g of resting cell paste of Gluconobacter oxydans CKDBM0901C deposit No. KCTC 12282BP, and add 1-deoxy-1- (2-hydroxyethyl amino)-. Conversion culture was performed for 96 hours at 25 ° C. at 220 rpm using 100 g / of D-glue. Example 3. Synthesis of Migly from 6-hydroxyethyl amino-L-sorbose
실시예 2에서 CKDBM0901 휴면세포 페이스트 및 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D-글루시틀 100 g/ 를 사용한 전환배양을 실시하여 얻은 배양액 300 in£(50 m£w6)를 여과한 다음 활성탄을 처리하여 노란색 색소를 제거하고， 수소화 반응기 (PARR Instrument Co. USA, 5100 Reactor)로 이송하여 미글리를 합성을 수행하였다. 수소화 반웅을 위한 촉매로는 10% 팔라듐 -은-차콜을 사용하였고， 온도 25°C와 10기압의 조건에서 5시간 동안 반웅을 진행한 다음 여과하여 촉매를 제거하였다. 상기 여과액을 NMR법에 의하여 분석한 결과， 6-하이드록시에틸 아미노 -L- 소르보스로부터 미글리를이 합성되었음을 확인하였다. 실시예 4. 미글리를 샘플 제조 및 물질동정 300 in £ (50 m £ w6 culture medium obtained by conversion culture with CKDBM0901 dormant cell paste and 1 g of 1-deoxy-1- (2-hydroxyethyl amino) -D-glucitelle in Example 2 ), And then treated with activated carbon to remove the yellow pigment, and transferred to a hydrogenation reactor (PARR Instrument Co. USA, 5100 Reactor) to synthesize the Migley. 10% palladium-silver-charcoal was used as a catalyst for hydrogenation reaction, and reaction was performed for 5 hours at a temperature of 25 ° C. and 10 atm, followed by filtration to remove the catalyst. The filtrate was analyzed by NMR, and it was confirmed that Migly was synthesized from 6-hydroxyethyl amino-L-sorbose. Example 4 Sample Preparation and Identification of Migley
실시예 3에서 얻은 반응여과액으로부터 미글리를을 얻기 위하여， 여과액을 강산성 양이온 수지 (Dowex 50X2-200)에 흡착시킨 다음 메탄올과 30% 암모니아 수용액으로 용리하고 농축하였다. 농축물을 메탄올을 사용하여 결정화와 재결정화 과정을 거쳐 건조한 다음 가스 크로마토그래피로 분석한 결과, 6-하이드록시에될 아미노 -L-소르보스로부터 62%수율로 순도 99.6%의 미글리를이 제조되었음을 확인하였다.  To obtain Migley from the reaction filtrate obtained in Example 3, the filtrate was adsorbed onto a strong acid cationic resin (Dowex 50X2-200), eluted with methanol and 30% aqueous ammonia solution and concentrated. The concentrate was crystallized and recrystallized using methanol, dried and analyzed by gas chromatography to obtain 99.6% pure Migly from 62% yield from amino-L-sorbose to be 6-hydroxyl. It was confirmed.
상기와 같이 제조된 샘플에 대하여 IR과 NMR을 통하여 구초해석을 한 결과， 미글리톨의 분자구조와 동일함을 확인하였다 (도 4 및 5). 이상으로 본 발명의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현 예일 뿐이며， 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.  As a result of the analysis of the herb through the IR and NMR for the sample prepared as described above, it was confirmed that the molecular structure of miglitol is the same (Figs. Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims

【특허청구범위] 【청구항 1】 에탄올아민을 포함하는 유기용매 하에서， 수소화붕소착물을 환원제로 사용하여 D-글루시를을 하기 화학식 1의 1-데옥시 -1-(2-하이드록시에틸 아미노 )-D-글루시를로 제조하는 것을 특징으로 하는， 1-데옥시 -1_(2- 하이드록시에틸 아미노) -D-글루시를의 제조방법 : [Claim 1] Claim 1] In an organic solvent containing ethanolamine, D-glucin is formed by using a borohydride complex as a reducing agent, and 1-deoxy-1- (2-hydroxyethyl amino of the formula (1) A process for preparing 1-deoxy-1_ (2-hydroxyethyl amino) -D-gluches, characterized in that it is prepared with
[화학식 1]  [Formula 1]
湖! 權  Ugh!權
【청구항 2] [Claim 2]
제 1 항에 있어서, 상기 수소화붕소착물은 수소화붕소나트륨 (NaBH4), 수소화붕소시아노나트륨 (NaCNBH3) 및 디메틸아미노보란 (BH4N(C¾)2)으로 구성된 군으로부터 선택되는 것을 특징으로 하는 제조방법. The method of claim 1, wherein the borohydride complex is selected from the group consisting of sodium borohydride (NaBH 4 ), sodium borohydride sodium (NaCNBH 3 ) and dimethylaminoborane (BH 4 N (C¾) 2 ). Manufacturing method.
【청구항 3】 [Claim 3]
제 1 항에 있어서， 상기 제조방법은 D-글루코스의 몰수 대비 1 내지 10 당량의 양으로 수소화붕소착물을 사용하는 것을 특징으로 하는 제조방법.  The method according to claim 1, wherein the preparation method uses boron hydride complexes in an amount of 1 to 10 equivalents based on the number of moles of D-glucose.
【청구항 4】 [Claim 4]
제 1 항에 있어서， 상기 유기용매는 암모늄염을 추가적으로 포함하는 것을 특징으로 하는 제조방법.  The method of claim 1, wherein the organic solvent further comprises an ammonium salt.
【청구항 5】 [Claim 5]
제 4 항에 있어서， 상기 제조방법은 D-글루코스의 몰수 대비 5 내지 10 당량의 양으로 암모늄염을 사용하는 것을 특징으로 하는 제조방법.  The method according to claim 4, wherein the production method uses ammonium salt in an amount of 5 to 10 equivalents relative to the number of moles of D-glucose.
【청구항 6】 [Claim 6]
제 4 항에 있어서， 상기 암모늄염은 아세트산 암모늄 (CH3C00NH4), 게 1인산 암모늄 (NH4¾P04), 제 2인산 암모늄 ((NH4)2HP04), 황산암모늄 ((NH4)2S04), 질산암모늄 (NH4M¾) 및 탄산암모늄 ((NH4)2C03)으로 구성된 군으로부터 선택되는 것을 특징으로 하는 제조방법. The method of claim 4, wherein the ammonium salt is ammonium acetate (CH 3 C00NH 4 ), ammonium monophosphate (NH 4 ¾P0 4 ), diammonium phosphate ((NH 4 ) 2 HP0 4 ), Process for the production of ammonium sulfate ((NH 4 ) 2 S0 4 ), ammonium nitrate (NH 4 M¾) and ammonium carbonate ((NH 4 ) 2 C0 3 ).
【청구항 7】 [Claim 7]
제 1 항에 있어서， 상기 유기용매는 알코올계 용매인 것을 특징으로 하는 제조방법 .  The method according to claim 1, wherein the organic solvent is an alcohol solvent.
【청구항 8】 [Claim 8]
제 1 항에 있어서, 상기 제조방법은 0 내지 40°C의 : 온도에서 실시되는 것을 특징으로 하는 제조방법. The method of claim 1, wherein the manufacturing method is carried out at a temperature of : 0 to 40 ° C.
【청구항 9】 [Claim 9]
다음의 단계를 포함하는 미글리틀 (Miglitol)의 제조방법:  Manufacturing method of miglitol comprising the following steps:
(a) 에탄을아민을 포함하는 유기용매 하에서， 수소화붕소착물을 환원제로 사용하여 D-글루시를을 하기 화학식 1의 1-데옥시 -1-(2- 하이드록시에틸 아미노) -D-글루시틀로 제조하는 단계;  (a) ethane is used as the reducing agent under an organic solvent containing amine, and D-glue is prepared by using 1-deoxy-1- (2-hydroxyethyl amino) -D-glu as shown in Formula 1 below. Manufacturing into a seattle;
[화학 1]  [Chemistry 1]
Figure imgf000018_0001
Figure imgf000018_0001
(b) 단계 (a)에서 제조한 1-데옥시 -1-(2-하이드록시에틸 아미노) -D- 글루시톨을 글루코노박터 속 균주 또는 이의 휴면세포 (resting cell)를 이용하여 6-하이드록시에틸 아미노 -L-소르보스로 생물전환 하는 단계; 및(b) 1-deoxy-1- (2-hydroxyethyl amino) -D-glucinol prepared in step (a) using a strain of Gluconobacter or its resting cell; Bioconversion to hydroxyethyl amino-L-sorbose; And
(c) 단계 (b)에서 생물전환 된 6-하이드록시에틸 아미노 -L- 소르보스를 미글리를로 합성하는 단계. (c) synthesizing 6-hydroxyethyl amino-L-sorbose biomigrated in step (b) into Migly.
PCT/KR2013/009599 2012-10-29 2013-10-25 Method for preparing 1-deoxy-1-(2-hydroxyethyl amino)-d-glucitol and miglitol WO2014069846A1 (en)

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CN115010610A (en) * 2022-06-16 2022-09-06 常州大学 Synthesis method of miglitol intermediate N-hydroxyethyl glucosamine

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* Cited by examiner, † Cited by third party
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CN105968042A (en) * 2016-07-15 2016-09-28 四川维奥制药有限公司 Preparation method of migltol
CN115010610A (en) * 2022-06-16 2022-09-06 常州大学 Synthesis method of miglitol intermediate N-hydroxyethyl glucosamine

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