CN101616888A - The purification process of 4-hydroxyisoleucine - Google Patents
The purification process of 4-hydroxyisoleucine Download PDFInfo
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Abstract
That problem of the present invention is to provide is easy, high purity, high yield separation and purification remove with separating (2S, 3R, 4S)-method of 4-hydroxyisoleucine.Particularly, the present invention openly comprise following operation (a) and (b) and (c) (2S, 3R, 4S)-4-hydroxyisoleucine or its purification process of acceptable salt chemically.Operation (a) the: make (2S in the mixture, 3R, 4S)-4-hydroxyisoleucine or its chemically acceptable salt and formula Q-CHO[formula in, it is that 6～14 aryl, carbonatoms are that 1～10 alkyl, carbonatoms are 3～10 cycloalkyl or 5～10 yuan heterocyclic radical that Q represents to have respectively substituent carbonatoms] aldehyde cpd or the reaction of its equivalents of expression, be converted into the compound [in the formula, Q as defined above] of formula (1) expression; Operation (b): the compound that with an organic solvent extracts formula (1) expression; And operation (c): the compound of formula (1) expression is converted into (2S, 3R, 4S)-4-hydroxyisoleucine or its chemically acceptable salt.
Description
Technical field
The present invention relates to the separation purification method of 4-hydroxyisoleucine, more specifically, relate to utilized to the aminal body transform (2S, 3R, 4S)-separation purification method of 4-hydroxyisoleucine with separate the method for removing.
Background technology
Known 4-hydroxyisoleucine is a kind of of nonprotein acidic amino acid, extracts from Fenugreek (Semen Trigonellae) (Trigonella foenum-graecum L.) seed etc. by (non-patent literature 1,2,3).
About this 4-hydroxyisoleucine; reported that the dependent insulin secretion of blood glucose value promotes glycogenesis in the enhancement (patent documentation 2,3, non-patent literature 5), muscle of active (patent documentation 1, non-patent literature 4), insulin sensitivity to promote the provide protection (patent documentation 5), fat absorbing restraining effect (patent documentation 6), anti-obesic action (patent documentation 7) of active (patent documentation 4, non-patent literature 6), skin or scalp etc., useful as the pharmaceuticals that utilize above-mentioned effect, heath food raw material, makeup (fragrant make-up product) raw material etc.And then it is also useful as the synthetic intermediate of various pharmaceuticals.
Now, as the method that obtains the 4-hydroxyisoleucine, known extraction (patent documentation 8,9, non-patent literature 1,2,3) and chemosynthesis (patent documentation 10,11, non-patent literature 7) from the Fenugreek seed.
In extraction method,, need carry out loaded down with trivial details operations such as chromatographic run or recrystallization with silica gel or ion exchange resin in order to separate with saponin class or other amino acid.The contained 4-hydroxyisoleucine of Fenugreek seed particularly, remove (the 2S that the viewpoint contain based on the above-mentioned effect of performance is considered to more important steric isomer, 3R, 4S)-the 4-hydroxyisoleucine beyond, also with the ratio that is about 9: 1 contain other steric isomers (2R, 3R, 4S)-4-hydroxyisoleucine (non-patent literature 2), even reuse chromatogram or recrystallization, also be not easy to separate aforementioned stereoisomers.
In the patent documentation 8, though chromatogram of being undertaken by spent ion exchange resin and recrystallization from the Fenugreek seed of 100g degreasing, obtain 0.6g purity be 99% (2S, 3R 4S)-the 4-hydroxyisoleucine, think that this rate of recovery is not high.Reason is particularly can't avoid the loss of target compound in filtrate owing to its characteristic when recrystallization, carries out its rate of recovery of recrystallization more and reduces more.
In the synthesis method,,, need the loaded down with trivial details operation of HPLC or recrystallization and so in order to remove above-mentioned impurity because side reaction etc. usually contain steric isomer.In the aforesaid operations with cost face or the device side problem that difficulty, yield reduce that maximizes.
As mentioned above, also exist problem in the purification process of existing 4-hydroxyisoleucine, therefore more effective the and easy method of expectation.
No. 587476 specification sheets of [patent documentation 1] Europe patent
[patent documentation 2] international brochure that discloses No. 2001/015689
[patent documentation 3] international brochure that discloses No. 2005/039626
[patent documentation 4] international brochure that discloses No. 2005/021596
[patent documentation 5] international brochure that discloses No. 2004/087091
[patent documentation 6] spy opens the 2006-89449 communique
No. 2006/0223884 specification sheets of [patent documentation 7] U.S. Patent application
No. 5470879 specification sheets of [patent documentation 8] United States Patent (USP)
[patent documentation 9] international brochure that discloses No. 2005/084323
[patent documentation 10] international brochure that discloses No. 2004/099120
[patent documentation 11] international brochure that discloses No. 2006/051000
[non-patent literature 1] Phytochemistry, vol.12,1707-1711,1973
[non-patent literature 2] Phytochemistry, vol.15,325,1976
[non-patent literature 3] Phytochemistry, vol.28,1835-1841,1989
[non-patent literature 4] Diabetes, Vol.47, Issue 2206-210,1998
[non-patent literature 5] American Journal of Physiology Endocrinology andMetabolism, Vol.287, E463-E471,2004
[non-patent literature 6] Amino Acids, vol.28,71-76,2005
[non-patent literature 7] European Joumal of Organic Chemistry, vol.5,834-839,2002
Summary of the invention
Invent problem to be solved
The object of the present invention is to provide easy, high purity and separation and purification with high yield (2S, 3R, 4S)-4-hydroxyisoleucine or its chemically acceptable salt (followingly be also referred to as sometimes " (2S, 3R, 4S)-HIL ") method.
In addition, the present invention also aims to provide from mixture and to separate easily and remove (2S, 3R, 4S)-method of 4-hydroxyisoleucine.
Solve the method for problem
The inventor finds to pass through with (2S, 3R, 4S)-after the 4-hydroxyisoleucine is converted into the aminal body, carry out organic solvent extraction again, can from its steric isomer come from the saponin class of crude extract or other compounds such as other amino acid easy and separate this aminal body etc. well, thereby finished the present invention.
That is, the present invention is as described below.
(1) (4S)-4-hydroxyisoleucine or its purification process of acceptable salt chemically, wherein, described method comprises following operation (a) and (b) and (c) for 2S, 3R.
Operation (a) the: make (2S in the mixture, 3R, 4S)-4-hydroxyisoleucine or its chemically acceptable salt and formula Q-CHO[formula in, it is that 6～14 aryl, carbonatoms are that 1～10 alkyl, carbonatoms are 3～10 cycloalkyl or 5～10 yuan heterocyclic radical that Q represents to have substituent carbonatoms] aldehyde cpd or the reaction of its equivalents of expression, be converted into the compound of formula (1) expression
[in the formula, Q as defined above];
Operation (b): the compound that with an organic solvent extracts formula (1) expression; And
Operation (c): the compound of formula (1) expression is converted into (2S, 3R, 4S)-4-hydroxyisoleucine or its chemically acceptable salt.
(2) above-mentioned (1) described method, wherein, the aldehyde cpd that in the reaction of operation (a), uses formula Q-CHO to represent.
(3) above-mentioned (1) described method is characterized in that: by the abstraction process of operation (b), remove impurity from the compound of formula (1) expression.
(4) above-mentioned (1) or (2) described method, wherein, in the formula (1), Q is for can have substituent phenyl, cyclohexyl, naphthyl, pyridyl, thienyl, furyl or pyrryl.
(5) above-mentioned (4) described method, wherein, in the formula (1), Q is for can have substituent phenyl.
(6) above-mentioned (4) or (5) described method, wherein, this substituting group when Q has substituting group is selected from the group that is made of following radicals:
(i) carbonatoms be 1～6 alkyl,
(ii) carbonatoms be 1～6 alkoxyl group,
(iii) carbonatoms be 2～7 alkyloyl,
(iv) carbonatoms be 6～10 aryl,
(v) carbonatoms be 6～10 aryloxy,
(vi) halogeno-group,
(vii) carbonatoms be 1～6 haloalkyl,
(viii) nitro,
(ix) formyl radical and
(x) cyano group,
(wherein, above-mentioned (i) and (vii) not becoming carbonatoms and be the substituting group of 1～10 alkyl).
(7) above-mentioned (1) described method, wherein, the organic solvent of operation (b) is at least a alkyl acetate.
(8) above-mentioned (1) described method; wherein; in operation (c); use is selected from least a nucleophilic reagent in the group that is made of following material, and described material is a water; can be the azanol that 1～4 alkyl replaces by carbonatoms; ammonia; carbonatoms is 1～4 alkylamine; carbonatoms is 2～8 dialkylamine; can be selected from carbonatoms is 1～4 alkyl; carbonatoms is that 6～10 aryl and carbonatoms are the hydrazine that the substituting group in 2～7 the alkyloyl replaces; with by carbonatoms be the quaternary ammonium halides that 1～6 alkyl replaces; and their chemically acceptable salt.
(9) above-mentioned (1) described method, wherein, at least a alkyl acetate, carry out operation (c) to (2S, 3R, 4S)-4-hydroxyisoleucine or its chemically conversion of acceptable salt.
(10) above-mentioned (9) described method, wherein, at least a alkyl acetate is an ethyl acetate.
(11) (2S, 3R, 4S)-4-hydroxyisoleucine or its chemically the separation of acceptable salt remove method, wherein, described method comprises following operation (a) and (b).
Operation (a) the: make (2S in the mixture, 3R, 4S)-4-hydroxyisoleucine or its chemically acceptable salt and formula Q-CHO[formula in, it is that 6～14 aryl, carbonatoms are that 1～10 alkyl, carbonatoms are 3～10 cycloalkyl or 5～10 yuan heterocyclic radical that Q represents to have substituent carbonatoms] aldehyde cpd or the reaction of its equivalents of expression, be converted into the compound of formula (1) expression
[in the formula, Q as defined above];
Operation (b): the compound that with an organic solvent extracts formula (1) expression.
(12) above-mentioned (11) described method, wherein, the aldehyde cpd that in the reaction of operation (a), uses formula Q-CHO to represent.
(13) above-mentioned (11) or (12) described method, wherein, in the formula (1), Q is for can have substituent phenyl, cyclohexyl, naphthyl, pyridyl, thienyl, furyl or pyrryl.
(14) above-mentioned (13) described method, wherein, in the formula (1), Q is for can have substituent phenyl.
(15) above-mentioned (13) or (14) described method, wherein, this substituting group when Q has substituting group is selected from the group that is made of following radicals:
(i) carbonatoms be 1～6 alkyl,
(ii) carbonatoms be 1～6 alkoxyl group,
(iii) carbonatoms be 2～7 alkyloyl,
(iv) carbonatoms be 6～10 aryl,
(v) carbonatoms be 6～10 aryloxy,
(vi) halogeno-group,
(vii) carbonatoms be 1～6 haloalkyl,
(viii) nitro,
(ix) formyl radical and
(x) cyano group,
(wherein, above-mentioned (i) and (vii) not being the substituting group of 1～10 alkyl) as carbonatoms.
(16) above-mentioned (11) described method, wherein, the organic solvent of operation (b) is at least a alkyl acetate.
The invention effect
Method of the present invention as (2S, 3R, 4S)-chemically the separation purification method of acceptable salt is useful for 4-hydroxyisoleucine or its, can provide than chromatogram or recrystallization more effectively, high yield and highly purified (2S, 3R, 4S)-HIL.
More specifically, can be from the Fenugreek extracting solution, derive from the mixture that fermented liquid that culturing micro-organisms obtains etc. contains the 4-hydroxyisoleucine and separate effectively (2S, 3R, 4S)-HIL.In addition, can from the mixture that contains the 4-hydroxyisoleucine that obtains with chemosynthesis, separate effectively (2S, 3R, 4S)-HIL.
In addition, method of the present invention as (2S, 3R 4S)-that method is removed in the separation of HIL is also useful, can from the mixture of compound, remove effectively (2S, 3R, 4S)-HIL.The result can improve other compounds in the mixture, for example its steric isomer (2R, 3R, 4S)-4-hydroxyisoleucine or its chemically acceptable salt (followingly be also referred to as sometimes " (2R, 3R, 4S)-HIL ") purity.
Embodiment
Below, describe the present invention in detail.
In the method for the present invention, (2S, 3R, 4S)-the 4-hydroxyisoleucine can be the form of acceptable salt chemically.As this " chemically acceptable salt ", so long as bring dysgenic salt to get final product for method of the present invention, be not particularly limited, for example can enumerate and ammonia; Basic metal such as sodium, potassium; Alkaline-earth metal such as calcium, magnesium; Aluminium; Zinc; Organic amines such as triethylamine, thanomin, morpholine, tetramethyleneimine, piperidines, piperazine, dicyclohexyl amine; Basic aminoacids such as arginine, Methionin; Mineral acids such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, Hydrogen bromide; Acetate, citric acid, phenylformic acid, toxilic acid, fumaric acid, tartrate, succsinic acid, tannic acid, butyric acid, hibenzic acid, pounce on organic carboxyl acids such as acid, enanthic acid, capric acid, tea chloric acid, Whitfield's ointment, lactic acid, oxalic acid, amygdalic acid, oxysuccinic acid; The salt of organic sulfonic acids such as methylsulfonic acid, Phenylsulfonic acid, tosic acid etc.
In purification process of the present invention or the separation method, become the mixture of its object so long as contain (2S, 3R, 4S)-chemical mixture of HIL gets final product, to the composition of its containing ratio or other materials without any qualification.As the mixture of the object that becomes the inventive method, can enumerate fermented liquid that plant tissue extract, animal tissues's extract, culturing micro-organisms obtain, come from mixture of the reaction solution after the chemosynthesis etc.
In addition, that purification process of the present invention also goes for removing is purer (2S, 3R, 4S)-situation of small amount of impurities in the rough thing of HIL.
In the purification process of the present invention, as its as a result gained (2S, 3R 4S)-purity of HIL is as long as improve than the purity before the purifying, are not particularly limited, but in the optimal way of present method, provide pure in fact, for example use
1H-NMR (300MHz) do not observe when measuring impurity peaks (2S, 3R, 4S)-HIL.
The compound of formula of the present invention (1) expression is (2S, 3R, 4S)-and the aminal body of HIL, in the formula (1), it is that 6～14 aryl, carbonatoms are that 1～10 alkyl, carbonatoms are 3～10 cycloalkyl or 5～10 yuan heterocyclic radical that Q represents to have substituent carbonatoms.
As above-mentioned " can have substituent carbonatoms be 6～14 aryl " it " carbonatoms is 6～14 aryl ", can enumerate for example phenyl, naphthyl, anthryl, phenanthryl, xenyl etc., can preferably enumerate carbonatoms and be 6～10 aryl (for example phenyl, naphthyl, xenyl etc.), more preferably phenyl.
As above-mentioned " can have substituent carbonatoms be 1～10 alkyl " it " carbonatoms is 1～10 alkyl ", can enumerate carbonatoms and be 1～10 straight or branched alkyl, preferred carbonatoms is 1～6 straight or branched alkyl (for example methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, sec-butyl, isobutyl-, the tertiary butyl, amyl group, hexyl etc.).
As above-mentioned " can have substituent carbonatoms be 3～10 cycloalkyl " it " carbonatoms is 3～10 cycloalkyl ", can enumerate for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl, ring octyl group, ring nonyl, ring decyl etc., can preferably enumerate carbonatoms and be 5～8 cycloalkyl (for example cyclopentyl, cyclohexyl, suberyl, ring octyl group etc.), more preferably cyclohexyl.
As above-mentioned " heterocyclic radical that can have 5～10 yuan of substituting groups " they " heterocyclic radicals of 5～10 yuan ", can enumerate except that carbon atom, also contain 1～4 heteroatomic 5～10 yuan of aromatic heterocycles or non-aromatic heterocycle and condensed ring radical thereof that is selected from Sauerstoffatom, sulphur atom and the nitrogen-atoms.
As this aromatic heterocyclic radical or its condensed ring radical, can enumerate 2-or 3-thienyl, 2-or 3-furyl, 1-, 2-or 3-pyrryl, 1-, 2-, 4-or 5-imidazolyl, 2-, 4-or 5-oxazolyl, 2-, 4-or 5-thiazolyl, 1-, 3-, 4-or 5-pyrazolyl, 3-, 4-or 5-isoxazolyl, 3-, 4-or 5-isothiazolyl, 1,2,4-triazole-1-, 3-, 4-or 5-base, 1,2,3-triazole-1-, 2-or 4-base, 1H-tetrazolium-1-or 5-base, 2H-tetrazolium-2-or 5-base, 2-, 3-or 4-pyridyl, 2-, 4-or 5-pyrimidyl, 1-, 2-, 3-, 4-, 5-, 6-or 7-indyl, 2-, 3-, 4-, 5-, 6-or 7-benzofuryl, 2-, 3-, 4-, 5-, 6-or 7-benzothienyl, 1-, 2-, 4-, 5-, 6-or 7-benzimidazolyl-, 2-, 3-, 4-, 5-, 6-, 7-or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-or 8-isoquinolyl etc.
Heterocyclic radical or its condensed ring radical as this non-aromatic, can enumerate 2-or 3-dihydro-thiophene base, 2-or 3-dihydrofuran base, 1-, 2-or 3-pyrrolinyl, 1-, 2-, 4-or 5-imidazolinyl, 2-, 4-or 5-oxazolinyl, 2-, 4-or 5-thiazolinyl, 1-, 3-, 4-or 5-pyrazolinyl, 3-, 4-or 5-isoxazoline-3-yl, 3-, 4-or 5-isothiazoline base, 1,2,4-triazoline-1-, 3-, 4-or 5-base, 1,2,3-triazoline-1-, 2-or 4-base, 1H-tetrazolium quinoline-1-or 5-base, 2H-tetrazolium quinoline-2-or 5-base, 2-, 3-or 4-dihydropyridine base, 2-, 4-or 5-dihydro-pyrimidin base, 1-, 2-, 3-, 4-, 5-, 6-or 7-indolinyl, 2-, 3-, 4-, 5-, 6-or 7-dihydro benzo furyl, 2-, 3-, 4-, 5-, 6-or 7-dihydrobenzo thienyl, 1-, 2-, 4-, 5-, 6-or 7-dihydrobenzo imidazolyl, 2-, 3-, 4-, 5-, 6-, 7-or 8-dihydroquinoline base, 1-, 3-, 4-, 5-, 6-, 7-or 8-dihydro-isoquinoline base, 2-or 3-tetrahydro-thienyl, 2-or 3-tetrahydrofuran base, 1-, 2-or 3-THP trtrahydropyranyl, 1-, 2-or 3-pyrrolidyl, 1-, 2-, 4-or 5-imidazolinyl, 2-, 4-or 5-oxazolinyl, 2-, 4-or 5-thiazolinyl, 1-, 3-, 4-or 5-pyrazolinyl, 3-, 4-or 5-isoxazoline-3-yl, 3-, 4-or 5-isothiazoline base, 1,2,4-triazoline-1-, 3-, 4-or 5-base, 1,2,3-triazoles quinoline-1-, 2-or 4-base, 1H-tetrazolium quinoline-1-or 5-base, 2H-tetrazolium quinoline-2-or 5-base, 2-, 3-or 4-piperidyl, 1-or 2-piperazinyl, 1-, 2-or morpholinyl, 1-, 2-or 3-thio-morpholinyl, 2-, 3-, 4-, 5-, 6-, 7-or 8-tetrahydric quinoline group, 1-, 3-, 4-, 5-, 6-, 7-or 8-tetrahydro isoquinolyl etc.
As this " heterocyclic radical ", thienyl, furyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, pyridyl, indyl, benzofuryl, benzothienyl, dihydro-thiophene base, dihydrofuran base, tetrahydro-thienyl, tetrahydrofuran base, THP trtrahydropyranyl etc. can be preferably enumerated, thienyl, furyl, pyridyl etc. can be more preferably enumerated.
In other embodiments, it is that 6～14 aryl, carbonatoms are 1～10 alkyl or 5～10 yuan heterocyclic radical that Q represents to have substituent carbonatoms.
As the substituting group of above-mentioned Q represented " can have substituent carbonatoms and be 6～14 aryl, carbonatoms and be 1～10 alkyl, carbonatoms and be 3～10 cycloalkyl or 5～10 yuan heterocyclic radical ", can enumerate for example following radicals:
(i) carbonatoms be 1～6 alkyl,
(ii) carbonatoms be 1～6 alkoxyl group,
(iii) carbonatoms be 2～7 alkyloyl,
(iv) carbonatoms be 6～10 aryl,
(v) carbonatoms be 6～10 aryloxy,
(vi) halogeno-group,
(vii) carbonatoms be 1～6 haloalkyl,
(viii) nitro,
(ix) formyl radical and
(x) cyano group,
(wherein, above-mentioned (i) and (vii) not becoming carbonatoms and be the substituting group of 1～10 alkyl).
In other embodiments, above-mentioned substituting group is selected from following radicals:
(i) carbonatoms be 1～6 alkyl,
(ii) carbonatoms be 1～6 alkoxyl group,
(iii) carbonatoms be 2～7 alkyloyl,
(iv) carbonatoms be 6～10 aryl,
(v) carbonatoms be 6～10 aryloxy,
(vi) halogeno-group,
(vii) carbonatoms be 1～6 haloalkyl,
(viii) nitro and
(ix) formyl radical,
(wherein, above-mentioned (i) and (vii) not becoming carbonatoms and be the substituting group of 1～10 alkyl).
Above-mentioned " (i) carbonatoms is 1～6 alkyl " is that carbonatoms is 1～6 straight or branched alkyl, for example methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, sec-butyl, isobutyl-, the tertiary butyl, amyl group, hexyl etc. be can enumerate, methyl, ethyl, n-propyl, sec.-propyl are preferably.
Above-mentioned " (ii) carbonatoms is 1～6 alkoxyl group " is the group that above-mentioned " carbonatoms is 1～6 alkyl " and Sauerstoffatom bonding obtain, for example methoxyl group, oxyethyl group, positive propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert.-butoxy, amyl group oxygen base, hexyl oxygen base etc. be can enumerate, methoxyl group, oxyethyl group, positive propoxy, isopropoxy are preferably.
Above-mentioned " (iii) carbonatoms is 2～7 alkyloyl " is the group that above-mentioned " carbonatoms is 1～6 alkyl " and carbonyl bonding obtain; for example ethanoyl, ethyl carbonyl, n-propyl carbonyl, sec.-propyl carbonyl, normal-butyl carbonyl, sec-butyl carbonyl, isobutyl-carbonyl, tertiary butyl carbonyl, amyl group carbonyl, hexyl carbonyl etc. be can enumerate, ethanoyl, ethyl carbonyl, n-propyl carbonyl, sec.-propyl carbonyl are preferably.
As above-mentioned " (iv) carbonatoms is 6～10 aryl ", can enumerate for example phenyl, naphthyl, xenyl etc., be preferably phenyl.
Above-mentioned " (v) carbonatoms is 6～10 aryloxy " be above-mentioned " carbonatoms is 6～10 aryl " and the group that the Sauerstoffatom bonding obtains, can enumerate for example phenoxy group, naphthyloxy, xenyl oxygen base etc., be preferably phenoxy group.
As above-mentioned " (vi) halogeno-group ", can enumerate for example fluorine atom, chlorine atom, bromine atoms, iodine atom etc., be preferably fluorine atom, chlorine atom, bromine atoms.
Above-mentioned " (vii) carbonatoms is 1～6 haloalkyl " be above-mentioned " carbonatoms is 1～6 the alkyl " quantity that can be replaced, be preferably the group that above-mentioned " halogeno-group " replacement more than a kind or a kind of 1～5 obtains, can enumerate for example methyl fluoride, difluoromethyl, trifluoromethyl, monochloro methyl, dichloromethyl, trichloromethyl, chlorine methyl fluoride, 1-fluoro ethyl, 1,1-two fluoro ethyls, 1,1,1-trifluoroethyl, pentafluoroethyl group, pentachloro-ethyl etc. are preferably trifluoromethyl, trichloromethyl.
Above-mentioned " carbonatoms is that 6～14 aryl, carbonatoms are that 1～10 alkyl, carbonatoms are 3～10 cycloalkyl or 5～10 yuan heterocyclic radical " is when having the substituting group that is selected from the above-mentioned substituting group (i)～(x), unless otherwise specified, unqualified to this substituent position and quantity.The quantity of above-mentioned substituting group (i)～(x) is 2 when above, and above-mentioned substituting group can be the same or different.But, above-mentioned (i) with (vii) do not become the substituting group of " carbonatoms is 1～10 alkyl ".
In addition, above-mentioned " carbonatoms is that 6～14 aryl, carbonatoms are 1～10 alkyl or 5～10 yuan heterocyclic radical " is when having the substituting group that is selected from the above-mentioned substituting group (i)～(ix), unless otherwise specified, this substituent position and quantity are not particularly limited.The quantity of above-mentioned substituting group (i)～(ix) is 2 when above, and above-mentioned substituting group can be the same or different.But, above-mentioned (i) with (vii) do not become the substituting group of " carbonatoms is 1～10 alkyl ".
As Q, preferably have to a certain degree volume and fat-soluble group, more specifically, preferably can have substituent phenyl, cyclohexyl, naphthyl, pyridyl, thienyl, furyl and pyrryl respectively, more preferably can have substituent phenyl.In above-mentioned group, when having substituting group should " substituting group ", can enumerate the substituting group that for example is selected from above-mentioned (i)～(x) etc.
In other embodiments, as Q, preferably can have substituent phenyl, naphthyl, pyridyl, thienyl, furyl and pyrryl respectively.
As Q, preferred especially nitrophenyl, phenyl, cyclohexyl.
In other embodiments, as Q, preferred especially nitrophenyl, phenyl.
Below, illustrate in greater detail method of the present invention.
Method of the present invention is based on that following understanding finishes, i.e. (2S, 3R, 4S)-HIL is converted into the compound of formula (1) expression, i.e. (2S well by the acetal amination reaction, 3R, 4S)-the aminal body (1) of HIL (below, also abbreviate " compound of formula (1) expression " or " aminal body (1) " sometimes as), and the compound of formula (1) is extracted in the organic solvent well.
For example, contain (2S, 3R, 4S)-when adding aldehyde cpd or its equivalents in the solution of HIL, generate (2S, 3R, 4S)-the aminal body (1) of HIL.Though (2S, 3R, 4S)-and HIL is difficult to directly extract in the organic solvent, and the fat-soluble raising of aminal body (1) that generates is extracted in the organic solvent easily.Thus, can be by extraction, separatory operation purifying aminal body (1).
On the other hand, as (2S, 3R, 4S)-steric isomer of HIL (2R, 3R, 4S)-HIL forms corresponding aminal body hardly in above-mentioned reaction.That is, use (2S, 3R, 4S)-HIL and (2R, 3R, 4S)-mixture of HIL, when under common condition, carrying out the acetal amination, only (2S, 3R, 4S)-optionally acetal amination of HIL.Thus, purification process of the present invention in addition also can remove with (2S, 3R, 4S)-the HIL structure on extremely similar compounds (2R, 3R, 4S)-HIL.
Below, describe each operation of method of the present invention in detail.
Operation (a)
In the method for the present invention, at first, in operation (a), will contain (2S, 3R, 4S)-mixture of HIL supplies in the acetal amination reaction.
The acetal amination reaction by make in the mixture (2S, 3R, 4S)-aldehyde cpd of HIL and formula Q-CHO (Q is as defined above) expression or its equivalents react and carry out.At this moment, do not use its equivalents, and preferably use aldehyde cpd itself.
Herein, the equivalents of so-called aldehyde cpd is meant the synthetic equivalents that plays a role equally with aldehyde cpd Q-CHO (Q as defined above) in this acetal amination reaction.As the equivalents of above-mentioned aldehyde cpd, the acetal body, hemiacetal body, aminal body, dihalo methyl body of the compound of representing corresponding to formula Q-CHO (Q as defined above) etc. are arranged, preferred especially acetal body and hemiacetal body.More specifically, can enumerate formula Q-CH (OR ') OR " acetal or the hemiacetal compound of [Q as defined above, R ' represents hydrogen atom, methyl or ethyl, R " expression methyl or ethyl, perhaps R ' and R " can bonding form ethylidene] expression.
The aldehyde cpd that formula Q-CHO represents or its equivalents are preferably added 1～5 equivalent with respect to the 4-hydroxyisoleucine content in the mixture.Herein, so-called " the 4-hydroxyisoleucine content in the mixture ", in the expression mixture (2S, 3R 4S)-content of HIL, contain in the mixture (2S, 3R, 4S)-HIL and its steric isomer (2S, 3R, 4S)-during HIL, represent their total content.
Above-mentioned reaction can be carried out in the presence of alkali as required.This alkali can be in mineral alkali or the organic bases any, mineral alkalis such as preference such as potassium hydroxide, sodium hydroxide, lithium hydroxide, yellow soda ash, sodium bicarbonate, organic basess such as triethylamine, diisopropylethylamine, morpholine, tetramethyleneimine, piperidines, piperazine, dicyclohexyl amine, thanomin, ammoniacal liquor etc.
The preferably about 0.1mol/L～2mol/L of alkali concn in the reaction solution is with respect to about preferably about 1～10 equivalent of 4-hydroxyisoleucine content in the mixture.
Above-mentioned reaction can be carried out in solvent as required.As this reaction solvent, preferably water, acetonitrile, acetone, tetrahydrofuran (THF), methyl alcohol, ethanol, Virahol or 1,4-diox or the mixed solvent that obtains with the arbitrary ratio mixing by above-mentioned solvent.
Preferred 0 ℃～50 ℃ of the temperature of reaction of above-mentioned reaction.
Operation (b)
Below, in operation (b), use the aminal body (1) of suitable organic solvent extraction gained.
As the organic solvent that extracts aminal body (1), for example can enumerate alkyl acetate, hexane, heptane, diethyl ether, t-butyl methyl ether, methylene dichloride, chloroform etc. not with any blended solvent of water and the mixture more than 2 kinds thereof, preferred at least a alkyl acetate.As this " alkyl acetate ", can enumerate the carbonatoms of acetate for example and be 1～10 alkyl ester, more preferably the carbonatoms of acetate is 1～4 alkyl ester (for example methyl acetate, ethyl acetate, n-propyl acetate, isopropyl acetate, n-butyl acetate, sec-butyl acetate, tert.-butyl acetate etc.).
At this moment, preferably can at organic solvent extraction aminal body (1) before post reaction mixture be added entry or alkali as required, make the aqueous solution of alkalescence, carry out organic solvent washing.Unreacted and residual aldehyde or its equivalents etc. can be removed by it being extracted in the organic solvent (for example alkyl acetate of above-mentioned explanation etc.).
If add acidic solution in the water layer after washing, make slightly acidic, the carboxyl that then can suppress aminal body (1) dissociates, and can be with suitable organic solvent extraction.Need to prove since this moment unreacted (2R, 3R, 4S)-HIL remains in the water layer, thus only (2S, 3R, 4S)-aminal body (1) the high purity ground of HIL extracts in the organic solvent, so can separate both effectively.
As the acidic solution that can adapt to, fat-soluble low aqueous acids such as preferred hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, Hydrogen bromide, acetate, citric acid, toxilic acid, fumaric acid, tartrate, succsinic acid, butyric acid, lactic acid, oxalic acid, oxysuccinic acid, methylsulfonic acid.
By aforesaid operation, can from aminal body (1), remove unreacted aldehyde or its equivalents, (2R, 3R, 4S)-impurity such as HIL.
By the aminal body (1) of organic solvent extraction proceed to following operation (c) take off aminal reaction before, can by partial crystallization, slurry washing, drying etc. usually used purification process be further purified.
Operation (c)
Below, in operation (c),, takes off in the aminal reaction aminal body (1) by being supplied to, be returned to (2S, 3R, 4S)-HIL.
The method of taking off the aminal reaction to aminal body (1) is not particularly limited, even for example only aminal body (1) is rested in the aqueous extraction solvent that is obtained by above-mentioned operation (b), takes off the acetal amination and also carries out, be converted into (2S, 3R, 4S)-HIL.
Preferred use nucleophilic reagent carries out this and takes off the aminal reaction.
As the nucleophilic reagent that can be fit to; preferably water; can be the azanol (for example azanol) that 1～4 alkyl replaces by carbonatoms; ammonia; carbonatoms is 1～4 alkylamine (for example methylamine); carbonatoms is 2～8 dialkylamine (for example dimethylamine); can be selected from carbonatoms is 1～4 alkyl; carbonatoms is that 6～10 aryl and carbonatoms are the hydrazine that replaces of the substituting group in 2～7 the alkyloyl (hydrazine for example; methylhydrazine; phenylhydrazine; acethydrazide); by carbonatoms is the quaternary ammonium halides that replaces of 1～6 alkyl (tetrabutyl ammonium fluoride for example; tetrabutylammonium chloride; Tetrabutyl amonium bromide; tetrabutylammonium iodide) etc., preferred especially azanol; methylamine and hydrazine.
In solvent, take off the aminal reaction as required.As this solvent, only otherwise bring detrimentally affect to get final product to reaction, can use any solvent, preference such as ester series solvent (for example with the explanation of above-mentioned operation (b) in the solvent phase enumerated with alkyl acetate), the mixed solvent of hydrocarbon system solvent (for example hexane, heptane), ether series solvent (for example diethyl ether, t-butyl methyl ether), halohydrocarbon series solvent (for example methylene dichloride, chloroform), pure series solvent (for example methyl alcohol, ethanol, Virahol) or above-mentioned solvent, more preferably the carbonatoms of at least a acetate is 1～4 alkyl ester, most preferably ethyl acetate.(2S, 3R, 4S)-solubleness of HIL in ethyl acetate is low, if suitably set concentration etc., then can obtain its crystallization easily as precipitate after reaction.
Need to prove, when particularly the Q in the formula (1) represents to have substituent carbonatoms and is the heterocyclic radical of 6～14 aryl or 5～10 yuan, preferably take off the aminal reaction by shortening.
As the metal catalyst that can adapt to this shortening, preferred palladium, rhodium, platinum etc.
As the reaction solvent that can adapt to this shortening, preferably water, methyl alcohol, ethanol or ethyl acetate or their mixed solvent.
Regressive as mentioned above (2S, 3R, 4S)-HIL can by heavy partial crystallization, slurry washing, drying etc. usually used purification process be further purified.
As mentioned above separation and purification (2S, 3R, 4S)-HIL is useful as pharmaceuticals (Remedies for diabetes etc.), heath food raw material, cosmetic material etc., and also useful as the synthetic intermediate of various pharmaceuticals.
For example, with (2S, 3R, 4S)-when HIL is used as diabetes mellitus prevention and/or therapeutical agent, can be directly or according to itself known method, make the pharmaceutical composition that is mixed together with pharmaceutically acceptable carrier and obtains, come administration by oral or non-oral (for example approach such as intravenously, subcutaneous, intramuscular, suppository, rectal injection, ointment, patch, hypogloeeis, eye drip, suction).The dosage that is used for above-mentioned purpose is according to determining as the result of treatment of target, medication, treatment time, age, body weight etc., but by oral or non-oral approach, dosage as common per day for adults, under case of oral administration with 1 μ g～10g, under non-case of oral administration with 0.01 μ g～1g, administration 1 day 1 time～for several times.In addition, in the aforementioned pharmaceutical compositions (2S, 3R, 4S)-content of HIL is about 0.01% (weight)～100% (weight) of composition in its entirety.
As the pharmaceutically acceptable carrier in the pharmaceutical composition, as preparation raw material, habitual various organic or inorganic carrier substance can be enumerated, vehicle, lubricant, tackiness agent, disintegrating agent, water-soluble polymer, alkaline, inorganic salts in the solid preparation for example can be enumerated; Solvent in the aqueous preparation, solubility promoter, suspensoid, isotonic agent, buffer reagent, painlessization agent etc.In addition, as required, also can use additives such as common sanitas, antioxidant, tinting material, sweeting agent, acidic flavoring agent, whipping agent, spices.
Agent shape as aforementioned pharmaceutical compositions, can enumerate for example tablet, powder, pill, granule, capsule, suppository, liquid preparation, sugar-coat agent, prolonged action preparation, syrup, suspensoid, emulsion, sugar-tablet agent, hypogloeeis agent, alite paste, intraoral disintegration agent (sheet), inhalation, rectal injection agent, ointment, patch agent, adhesive tape agent, eye drops, can use common formulation auxiliary agents to make according to conventional methods.
Aforementioned pharmaceutical compositions can wait according to the method for customary way, the record of for example Japanese Pharmacopoeia in the preparation technique field and make.Below, the concrete manufacture method of detailed description preparation.
For example, with (2S, 3R, 4S)-when HIL is modulated into preparations for oral administration, after adding vehicle and tackiness agent as required, disintegrating agent, lubricant, tinting material, drug flavoring etc., make for example tablet, powder, pill, granule, capsule, suppository, solution, sugar-coat agent, prolonged action preparation or syrup etc. by ordinary method.As vehicle, can enumerate for example lactose, W-Gum, sucrose, glucose, Sorbitol Powder, crystalline cellulose etc., as tackiness agent, can enumerate for example polyvinyl alcohol, polyvingl ether, ethyl cellulose, methylcellulose gum, gum arabic, tragakanta, gelatin, shellac, hydroxypropylcellulose, hydroxypropylated starch, polyvinylpyrrolidone etc., as disintegrating agent, can enumerate for example starch, agar, the gelatin powder, crystalline cellulose, lime carbonate, sodium bicarbonate, citrate of lime, dextrin, pectin etc., as lubricant, can enumerate for example Magnesium Stearate, talcum, polyoxyethylene glycol, silicon-dioxide, the hydrogenated vegetable wet goods, as tinting material, can use the tinting material that allows to add in the pharmaceuticals, as drug flavoring, can use cocoa powder, mentha camphor, aromatic acid, spearmint oil, borneol, Cortex cinnamomi japonici powder etc.Certainly above-mentioned tablet or granule are wrapped up sugar-coat, gelatin clothing and carry out suitably other dressing as required.
Modulation is added pH regulator agent, buffer reagent, stablizer, preservatives etc. during injection as required, makes subcutaneous, intramuscular, intravenous injection agent according to conventional methods.
In addition, as mentioned above, (2S, 3R, 4S)-HIL can as diabetes prevent and/or treat agent, also can unite use with the agent that prevents and/or treats of normally used other Remedies for diabetes, diabetic complication.So-called used Remedies for diabetes usually, diabetic complication prevent and/or treat agent, can enumerate for example insulin preparation, insulin derivates, insulin-like action agent, insulin secretion stimulators, agent for improvement of insulin resistance, the biguanides preparation, the glyconeogenesis inhibitor, the sugar absorption inhibitor, kidney sugar reuptake inhibithors, β 3 adrenoceptor agonists, glucagon-like-peptide-1 (7-37), glucagon-like-peptide-1 (7-37) analogue, glucagon-like peptide-1 receptor stimulant, inhibitors of dipeptidyl IV, aldose reductase inhibitor, the glycation end product formation inhibitor, glycogen synthase kinase-3 Depressant, the glycogen phosphorylase Depressant, antihyperlipidemic drug, appetite-inhibiting agent, the lipase Depressant, hypotensive agent, peripheral circulation is improved medicine, antioxidant, the combination more than a kind or 2 kinds or the mixture of diabetic neuropathy curative etc.
With (2S, 3R, 4S)-medicament that HIL is used in combination can be mixed and made into unitary agent, perhaps with other method with its preparationization respectively, or in a container will with other method with its respectively the material that obtains of preparationization carry out the combination preparation (group, box, bag etc.) of bale packing.
Administration form during to drug combination is not particularly limited, and for example can enumerate (1) as administration when the administration of unitary agent, (2) different preparations are under same route of administration, the administration when the free poor administration under the same route of administration, (4) different preparations are under different way of administration of (3) each preparation, the free poor administration under different way of administration of (5) each preparation etc.
Preferred disease about the particular compound of the medicament that is used in combination maybe should be disposed be listed below, but content of the present invention is not limited to this.
As insulin preparation, the Regular Insulin that can enumerate NPH, Lente, Ultralente, can absorb through lung etc.
So-called insulin derivates is meant by Regular Insulin deutero-protein or peptide, keeps the material of insulin action, can enumerate for example to rely dried meat (Lispro), B10Asp, glargine etc.
So-called insulin-like action agent, be meant and do not rely on the physiological action that the performance of Regular Insulin ground promotes the Regular Insulin such as effect of cellular uptake sugar to a certain extent, bring into play the insulin derivates preparation in addition of hypoglycemic activity thus, can enumerate for example insulin receptor kinase excitants (for example L-783281, TER-17411, CLX-0901, KRX-613 etc.), vanadium etc.
So-called insulin secretion stimulators; be meant and act on pancreatic beta cell; the insulin secretion of increase in blood; bring into play the preparation of hypoglycemic activity thus, can enumerate ATP sensitive potassium channel inhibitor beyond for example sulfonylurea agent (for example tolbutamide, P-607, tolazamide, acetohexamide, gliclazide, glimepiride, Glipizide, Glyburide (Glyburide) etc.), meglitinide class (for example nateglinide, repaglinide, mitiglinide etc.), sulfonylurea agent and/or the meglitinide class (for example BTS-67-582 etc.) etc.
So-called agent for improvement of insulin resistance, be meant the preparation of bringing into play hypoglycemic activity by the insulin action in the destination organization that strengthens Regular Insulin, for example can enumerate peroxisome propagation medicine activated receptor (PPAR) gamma agonist (pioglitazone for example, rosiglitazone, troglitazone, thiazolidinedione based compounds such as ciglitazone, or GI-262570, GW-1929, JTT-501, non-thiazolidinedione based compound such as YM-440 etc.), PPAR γ antagonist (bisphenol A diglycidyl ether for example, LG-100641 etc.), PPAR alfa agonists (chlorine Bei Te, bezafibrate, Fibrate based compounds such as S-8527, or non-Fibrate based compound etc.), PPAR α/gamma agonist (for example KRP-297 etc.), retinoids X receptor stimulant (for example LG-100268 etc.), retinoids X receptor antagonist (for example HX531 etc.), Protein-tyrosine-phosphatase-1B inhibitor (for example PTP-112 etc.) etc.
So-called biguanides agent, be meant by anaerobic in glyconeogenesis restraining effect in the liver or the tissue and separate insulin resistant property improvement effect in sugared promoter action or the periphery etc., the preparation of performance hypoglycemic activity can be enumerated for example N1,N1-Dimethylbiguanide, phenformin, buformin etc.
So-called glyconeogenesis inhibitor is meant mainly by suppressing the preparation of glyconeogenesis performance hypoglycemic activity, for example can enumerate glucagon secretion inhibitor (M﹠amp for example; B 39890A etc.), glucagon receptor antagonist (for example CP-99711, NNC-92-1687, L-168049, BAY27-9955 etc.), G-6-Pase inhibitor etc.
So-called sugared absorption inhibitor, be meant that suppressing or postpone sugar by the contained enzymic digestion of carbohydrate in digestive tube in the inhibition food absorbs in the body, bring into play the preparation of hypoglycemic activity thus, can enumerate for example alpha-glucosidase inhibitor (for example acarbose, voglibose, miglitol etc.), alpha-amylase inhibitor (for example AZM-127 etc.) etc.
So-called kidney sugar reuptake inhibithors is meant the absorption again that suppresses sugar in the uriniferous tubules, brings into play the preparation of hypoglycemic activity thus, for example can enumerate sodium dependent glucose transporter inhibitors (for example T-1095, phlorizin etc.) etc.
So-called β 3 adrenoceptor agonists, be meant that β 3 adrenoceptors that stimulate in the adipocyte make Fatty Acid Oxidation hyperfunction, consumed energy is brought into play the preparation of the improvement effect of obesity, hyperinsulinemia thus, can enumerate for example CL-316243, TAK-677 etc.
As glucagon-like-peptide-1 (7-37) analogue, can enumerate for example acetic acid Exenatide (exendin-4), NN-2211 etc., as glucagon-like peptide-1 receptor stimulant, can enumerate for example AZM-134 etc., as inhibitors of dipeptidyl IV, can enumerate for example NVP-DPP-728 etc.Glucagon-like-peptide-1 analog, glucagon-like peptide-1 receptor stimulant, inhibitors of dipeptidyl IV and glucagon-like-peptide-1 are the effects by the glucagon-like-peptide-1 in simulation or the enhancing cell, the preparation of performance diabetes improvement effect.
So-called aldose reductase inhibitor, be meant in the material that is preferred for handling the diabetic complication, reduce the Sorbitol Powder of being confirmed in the tissue that the diabetic complication takes place in the hyperfunction and excessive cell of accumulating of polyvalent alcohol pathways metabolism that continues hyperglycemia state that results from, can enumerate for example epalrestat, tolrestatin, fidarestat, zenarestat etc. by suppressing aldose reductase.
So-called glycation end product formation inhibitor, be meant in the material that is preferred for disposing the diabetic complication, by suppress in the diabetic disease states because of continuing the material that a large amount of glycation end products that generate of hyperglycemia state alleviate the cell obstacle, above-mentioned diabetic complication prevents and/or treats agent and is also included within this category, preventing and/or treating agent with above-mentioned diabetic complication unites when using, except that above-mentioned diabetic complication prevents and/or treats agent, can enumerate for example NNC-39-0028, OPB-9195 etc.
As glycogen synthase kinase-3 Depressant, can enumerate for example SB-216763, CHIR-98014 etc., as the glycogen phosphorylase Depressant, can enumerate for example CP-91149 etc.
As antihyperlipidemic drug, for example can enumerating, hydroxymethyl glutaryl CoA reductase inhibitor (for example Pravastatin, Simvastatin, fluvastatin, atorvastatin etc.), Fibrate are that medicament (for example chlorine Bei Te, bezafibrate, simfibrate etc.), bile acide are drained promotion medicine etc.
As appetite suppressant, can enumerate for example sibutramine, Mazindol etc., as the lipase Depressant, can enumerate orlistat etc.
As hypotensive agent, can enumerate for example angiotensin-converting enzyme inhibitor (for example captopril, alacepril etc.), angiotensin-ii receptor antagonistic (for example candesartan cilexetil, valsartan etc.), calcium antagonists (for example cilnidipineb, amlodipine, nicardipine etc.), hydragog(ue) (for example trichlormethiazide, antisterone etc.), sympathetic nerve blocking agent (for example clonidine, serpentine etc.) etc.
Improve medicine as peripheral circulation, can enumerate for example EPA-EE etc.
As antioxidant, can enumerate polyprotonic acid, probucol etc.
As the diabetic neuropathy curative, can enumerate for example mecobalamin, mexiletine hydrochloride etc.
In addition, (2S, 3R, 4S)-HIL also is useful as food additives.
Contain (2S, 3R, 4S)-food compositions of HIL is useful as preventing and/or treating with food of diabetes.
" food " used in this specification sheets is meant all food, except that comprising so-called heath food, also comprises the food specific for health care or the trophic function food of the health functional food system defined of MHLW, also comprises dietary supplements.
Form to the food compositions put down in writing in this specification sheets is not particularly limited, so long as form that can orally ingestible can be any form.
For example, can enumerate powder, particle, tablet, hard capsule, soft capsule, liquid (beverage, jellylike drinks etc.), candy, chocolate etc., all can be by making in this technical field from the body known method.
In the food compositions (2S, 3R, 4S)-content of HIL suitably is defined as the suitable consumption of indicated scope.
Above-mentioned food compositions can use other foodstuff additive as required.As above-mentioned foodstuff additive, can enumerate the fruit juice of adjusting improved taste, dextrin, the ring-type oligosaccharides, carbohydrate (fructose, monose and polyoses such as glucose), acid flavoring, spices, smear tea powder etc., and the emulsifying agent that improves quality, collagen, milk powder, thickening polyose or agar etc., and vitamins, eggshell calcium, calcium pantothenate, other mineral substances, royal jelly, propolis, honey, foodstuff fibre, Larch agaric, chitin, chitosan, flavonoid, carotenoids, xenthophylls, herbal medicine, chrondroitin, each seed amino acid etc. are as the material of the composition of common heath food etc.
[embodiment]
Below, by embodiment the present invention is described in further detail, but the present invention is not limited to these embodiment.
(test example)
[analysis condition]
(1)
1H-NMR: measure with Varian Mercury 300VX (300MHz).
(2)LCMS：
(HPLC)：Hewlett?Packard?series?1100
Post: Symmetry (registered trademark) C183.5 μ m 4.6 * 50mm
Moving phase: the water and the acetonitrile that contains 0.04%TFA that contain 0.05%TFA
Gradient: the acetonitrile that contains 0.04%TFA is to be increased to 98% condition in 5 minutes from 5% straight line
Flow velocity: 2ml/ minute
Column temperature: 30 ℃
Detect: the UV detector (210nm～300nm)
(mass spectrum): use the Waters Micromass platform of company to detect ESI and PDA.
Embodiment 1: and purifying from the Fenugreek seed (2S, 3R, 4S)-HIL
The extraction of the rough thing of (operation 1) 4-hydroxyisoleucine (the rough thing of HIL)
With grinding 5g Fenugreek seed is ground, with the degreasing of 50ml hexane.Then, flood a night with 50ml 70% aqueous ethanolic solution after, leach residue, with 60ml 70% washing with alcohol.Filtrate is heated up in a steamer in decompression, obtains the rough thing of HIL.
Acetal amination and organic solvent extraction that (operation 2) carries out with paranitrobenzaldehyde
Rough thing of the above-mentioned HIL that obtains and 10ml 5% sodium bicarbonate aqueous solution are mixed, add the 3ml acetonitrile solution of 0.2g paranitrobenzaldehyde, stir a night down at 25 ℃.After acetonitrile is heated up in a steamer in decompression, with 50ml ethyl acetate (Kanto Kagaku K. K. 1 grade more than 99%) the excessive aldehyde of washing.The water layer of gained is neutralized with the 1M aqueous citric acid solution, use ethyl acetate extraction.Ethyl acetate layer with after the saturated common salt water washing, is used anhydrous magnesium sulfate drying, leaches this siccative, obtain the aminal body, (2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-nitrophenyl)-tetrahydrochysene-2H-1, the ethyl acetate solution of 3-oxazine-4-formic acid.
MS(ESI)m/z?281[M+H]
+
(operation 3) takes off the acetal amination
(the 2R that obtains in above-mentioned operation, 4S, 5R, 6S)-5,6-dimethyl-2-(4-nitrophenyl)-tetrahydrochysene-2H-1, add 0.1ml 50% aqueous hydroxylamine in the ethyl acetate solution of 3-oxazine-4-formic acid, left standstill 3 days, the crystallization that leaching is separated out obtains 30mg (2S, 3R, 4S)-the 4-hydroxyisoleucine.In this crystallization, confirm as unimodally with LCMS, and then promptly use
1H-NMR does not see impurity peaks yet, confirm as highly purified (2S, 3R, 4S)-HIL.
1H-NMR(300MHz，D
2O)δ＝0.83(3H，d，J＝7.2Hz)，1.12(3H，d，J＝6.3Hz)，1.75-1.83(1H，m)，3.70(1H，m)，3.76(1H，d，J＝7.8Hz)
MS(ESI)m/z?148[M+H]
+
Advancing of embodiment 2:4-hydroxyisoleucine stereoisomer mixture with paranitrobenzaldehyde
The acetal amination of row ((2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-nitrophenyl)
-four oxygen-2H-1,3-oxazine-4-formic acid synthetic)
With 0.4g (2S, 3R, 4S)-the 4-hydroxyisoleucine with (2R, 3R, 4S)-about 1: 1 mixture and 10ml 5% sodium bicarbonate aqueous solution of 4-hydroxyisoleucine mix, and adds the 20ml acetonitrile solution of 0.41g paranitrobenzaldehyde, stirs a night.After acetonitrile is heated up in a steamer in decompression, with the excessive aldehyde of ethyl acetate washing.The water layer of gained is neutralized with the 1M aqueous hydrochloric acid, use ethyl acetate extraction.After the water washing of ethyl acetate layer usefulness saturated common salt, use anhydrous magnesium sulfate drying.After leaching this siccative, explosive decompression heats up in a steamer and desolvates, obtain 0.3g (2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-nitrophenyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid (corresponding to (and 2S, 3R, 4S)-steric isomer of HIL).In the solid of gained, confirm as unimodally with LCMS, and then promptly use
1H-NMR do not see yet from corresponding to (2R, 3R, 4S)-peak of the steric isomer of HIL.In addition, the absolute configuration of the compound of gained is also supported by its NOESY spectrum.
1H-NMR(300MHz，CD
3OD)δ＝1.00(3H，d，J＝6.6Hz)，1.34(3H，d，J＝6.0Hz)，1.45-1.58(1H，m)，3.47(1H，d，J＝10.5Hz)，3.60-3.70(1H，m)，5.39(1H，s)，7.76(2H，d，J＝8.7Hz)，8.23(2H，d，J＝8.7Hz)
MS(ESI)m/z?281[M+H]
+
Measure NOESY spectrographic result, observe following intersection peak.
That is, the compound of confirming gained be have following steric configuration (2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-nitrophenyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid.
Embodiment 3: the aminal that takes off that carries out with azanol reacts
To carry out 50mg (0.18mmol) (2R, 4S, the 5R that same operation obtains with embodiment 2,6S)-5,6-dimethyl-2-(4-nitrophenyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid is dissolved in the 2ml ethyl acetate, add 0.01ml 50% aqueous hydroxylamine, stirred 2 hours.The crystallization that leaching is separated out, obtain 7mg (2S, 3R, 4S)-the 4-hydroxyisoleucine.In the crystallization of gained, confirm as unimodally with LCMS, and then promptly use
1H-NMR does not see impurity peaks yet, confirm as highly purified (2S, 3R, 4S)-the 4-hydroxyisoleucine.
1H-NMR(300MHz，D
2O)δ＝0.83(3H，d，J＝7.2Hz)，1.12(3H，d，J＝6.3Hz)，1.75-1.83(1H，m)，3.70(1H，m)，3.76(1H，d，J＝7.8Hz)
MS(ESI)m/z?148[M+H]
+
Embodiment 4: the aminal that takes off that carries out with methylamine methanol solution reacts
Use 0.014ml 40% methylamine methanol solution to replace 50% aqueous hydroxylamine, operate based on embodiment 3, can obtain thus 4mg (2S, 3R, 4S)-HIL.The crystallization of gained is confirmed as unimodal with LCMS, and then promptly uses
1H-NMR does not see impurity peaks yet, confirm as highly purified (2S, 3R, 4S)-the 4-hydroxyisoleucine.
Embodiment 5: the aminal that takes off that carries out with the hydrazine monohydrate reacts
Use 0.01ml hydrazine monohydrate to replace 50% aqueous hydroxylamine, operate based on embodiment 3, can obtain 20mg (2S, 3R, 4S)-HIL.In the crystallization of gained, confirm as unimodally with LCMS, and then promptly use
1H-NMR does not see impurity peaks yet, confirm as highly purified (2S, 3R, 4S)-the 4-hydroxyisoleucine.
Carrying out of embodiment 6:4-hydroxyisoleucine stereoisomer mixture with cyclohexyl aldehyde
The acetal amination ((2R, 4S, 5R, 6S)-and 2-cyclohexyl-5,6-dimethyl-tetrahydrochysene-
2H-1,3-oxazine-4-formic acid synthetic)
With 0.1g (0.68mmol) (2S, 3R, 4S)-HIL and (2R, 3R, 4S)-about 3: 2 mixtures of HIL are dissolved in 2.3ml 5% sodium bicarbonate aqueous solution, add the 2.3ml acetonitrile solution of 0.076g (0.68mmol) cyclohexyl aldehyde, stir a night.After acetonitrile is heated up in a steamer in decompression, with the excessive aldehyde of ethyl acetate washing.The water layer of gained is neutralized with the 2M aqueous citric acid solution, use ethyl acetate extraction.After the water washing of ethyl acetate layer usefulness saturated common salt, use anhydrous magnesium sulfate drying.After leaching siccative, explosive decompression heats up in a steamer and desolvates, obtain 0.067g (2R, 4S, 5R, 6S)-2-cyclohexyl-5,6-dimethyl-tetrahydrochysene-2H-1,3-oxazine-4-formic acid.
1H-NMR(300MHz，D
2O)δ＝1.06(3H，d，J＝6.6Hz)，1.07-1.40(6H，m)，1.60-2.00(9H，m)，3.34(1H，d，J＝11.4Hz)，3.42-3.50(1H，m)，4.24(1H，d，J＝6.6Hz)
MS(ESI)m/z?242[M+H]
+
Embodiment 7: with azanol carry out (2R, 4S, 5R, 6S)-2-cyclohexyl-5,6-two
Methyl-tetrahydrochysene-2H-1, the aminal that takes off of 3-oxazine-4-formic acid reacts
With the 0.067g (0.42mmol) of embodiment 6 gained (2R, 4S, 5R, 6S)-2-cyclohexyl-5,6-dimethyl-tetrahydrochysene-2H-1,3-oxazine-4-formic acid is dissolved in the 3ml methyl alcohol, adds 0.017ml 50% aqueous hydroxylamine, stirs 2 hours.Decompression adds ethyl acetate after heating up in a steamer and desolvating in the residue of gained, the insoluble crystallization of leaching, obtain 28mg (2S, 3R, 4S)-the 4-hydroxyisoleucine.In the crystallization of gained, confirm as unimodally with LCMS, promptly use
1H-NMR does not see impurity peaks yet, be confirmed to be highly purified (2S, 3R, 4S)-the 4-hydroxyisoleucine.
The acetal amination of carrying out with phenyl aldehyde of the stereoisomer mixture of embodiment 8:HIL
((2R, 4S, 5R, 6S)-5,6-dimethyl-2-phenyl-tetrahydrochysene-2H-1,3-
Synthesizing of oxazine-4-formic acid)
Acetal amination and organic solvent extraction that (operation 1) carries out with phenyl aldehyde
With 0.1g (2S, 3R, 4S)-HIL and (2R, 3R, 4S)-about 1: 1 mixture of HIL is dissolved in 2.7ml 5% sodium bicarbonate aqueous solution, adds the 2.7ml acetonitrile solution of 0.084g phenyl aldehyde, stirs a night.After acetonitrile is heated up in a steamer in decompression, with the excessive aldehyde of ethyl acetate (Kanto Kagaku K. K. 1 grade more than 99%) washing.The water layer of gained is neutralized with the 1M aqueous citric acid solution, use ethyl acetate extraction.After the water washing of ethyl acetate layer usefulness saturated common salt, use anhydrous magnesium sulfate drying.Leach siccative, obtain (2R, 4S, 5R, 6S)-5,6-dimethyl-2-phenyl-tetrahydrochysene-2H-1, the ethyl acetate solution of 3-oxazine-4-formic acid.
(operation 2) reacts with the aminal that takes off that shortening carries out
With ethyl acetate layer leave standstill at room temperature, in the open system, with separate out the crystalline form obtain the 12mg high purity (
1H-NMR and LCMS) (2S, 3R, 4S)-HIL.And then, remove this crystallization by leaching after, with the aminal body in the filtrate according to operation shown below, be converted into (2S, 3R, 4S)-HIL.
The solvent in the filtrate is heated up in a steamer in decompression, adds 1: 2 mixed solvent of entry and methyl alcohol, 10% palladium carbon of catalytic amount in residue, replaces system with hydrogen, stirs a night.After leaching palladium carbon, by concentrating under reduced pressure and freeze-drying, obtain 40mg (2S, 3R, 4S)-HIL.This dried frozen aquatic products is confirmed as unimodal with LCMS.
Embodiment 9: the acetal amination of carrying out with various aldehyde
According to obtaining following compound with the embodiment 6 identical method of putting down in writing.
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-phenyl-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
1H-NMR(300MHz，CD
3OD)δ＝1.09(3H，d，J＝6.6Hz)，1.35(3H，d，J＝6.3Hz)，1.63-1.78(1H，m)，3.53(1H，d，J＝10.8Hz)，3.68-3.74(1H，m)，5.47(1H，s)，7.40-7.44(3H，m)，7.54-7.60(2H，m)
MS(ESI)m/z?236[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-pyridyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
1H-NMR(300MHz，CD
3OD)δ＝0.98(3H，d，J＝6.6Hz)，1.34(3H，d，J＝6.3Hz)，1.42-1.58(1H，m)，3.45(1H，d，J＝10.8Hz)，3.60-3.68(1H，m)，5.31(1H，s)，7.59(2H，d，J＝4.8Hz)，8.54(2H，d，J＝4.8Hz)
MS(ESI)m/z?237[M+H]
+
(2R, 4S, 5R, 6S)-and 2-(4-chloro-phenyl-)-5,6-dimethyl-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?270[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-p-methoxy-phenyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?266[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-aminomethyl phenyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?250[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-trifluoromethyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?304[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(4-fluorophenyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?254[M+H]
+
(2R, 4S, 5R, 6S)-and 2-(4-cyano-phenyl)-5,6-dimethyl-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?261[M+H]
+
(2R, 4S, 5R, 6S)-and 2-(2-chloro-phenyl-)-5,6-dimethyl-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?270[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(2-nitrophenyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?281[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(3-nitrophenyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?281[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(2-thienyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?242[M+H]
+
(2R, 4S, 5R, 6S)-5,6-dimethyl-2-(2-furyl)-tetrahydrochysene-2H-1,3-oxazine-4-formic acid
MS(ESI)m/z?226[M+H]
+
Utilizability on the industry
The method according to this invention, can be effectively and obtain with high yield highly purified (2S, 3R, 4S)-4-HIL. (2S, 3R, the 4S)-4-HIL of gained is as for example pharmaceuticals (Remedies for diabetes etc.), healthy food raw material, cosmetic material etc. are useful, and then as each The synthetic intermediate of planting pharmaceuticals is also useful.
In addition, method of the present invention is removed method also as the separation of (2S, 3R, 4S)-HIL Useful, can from the mixture of compound, effectively remove (2S, 3R, 4S)-HIL.
Japanese patent application 2007-042711 and the Japanese patent application 2007-144578 of the application to propose in Japan, its content all comprises in this manual.In addition, patent documentation of quoting in this specification sheets and non-patent literature mode by reference discloses its full content, and with degree join in this specification sheets.
Claims (16)
1. (4S)-4-hydroxyisoleucine or its purification process of acceptable salt chemically, wherein, described method comprises following operation (a) and (b) and (c) for 2S, 3R,
Operation (a) the: make (2S in the mixture, 3R, 4S)-aldehyde cpd or the reaction of its equivalents that 4-hydroxyisoleucine or its chemically acceptable salt and formula Q-CHO represent, be converted into the compound of formula (1) expression, in the formula, it is that 6～14 aryl, carbonatoms are that 1～10 alkyl, carbonatoms are cycloalkyl or 5 yuan of heterocyclic radical～10 yuan heterocyclic radicals of 3～10 that Q represents to have substituent carbonatoms
In the formula, Q as defined above;
Operation (b): the compound that with an organic solvent extracts formula (1) expression; And
Operation (c): the compound of formula (1) expression is converted into (2S, 3R, 4S)-4-hydroxyisoleucine or its chemically acceptable salt.
2. the described method of claim 1, wherein, the aldehyde cpd that in the reaction of operation (a), uses formula Q-CHO to represent.
3. the described method of claim 1 is characterized in that: by the abstraction process of operation (b), remove impurity from the compound of formula (1) expression.
4. claim 1 or 2 described methods, wherein, in the formula (1), Q is for can have substituent phenyl, cyclohexyl, naphthyl, pyridyl, thienyl, furyl or pyrryl.
5. the described method of claim 4, wherein, in the formula (1), Q is for can have substituent phenyl.
6. claim 4 or 5 described methods, wherein, this substituting group when Q has substituting group is selected from the group that is made of following radicals:
(i) carbonatoms be 1～6 alkyl,
(ii) carbonatoms be 1～6 alkoxyl group,
(iii) carbonatoms be 2～7 alkyloyl,
(iv) carbonatoms be 6～10 aryl,
(v) carbonatoms be 6～10 aryloxy,
(vi) halogeno-group,
(vii) carbonatoms be 1～6 haloalkyl,
(viii) nitro,
(ix) formyl radical and
(x) cyano group,
Wherein, above-mentioned (i) and (vii) do not become carbonatoms and be the substituting group of 1～10 alkyl.
7. the described method of claim 1, wherein, the organic solvent of operation (b) is at least a alkyl acetate.
8. the described method of claim 1; wherein; in operation (c); use is selected from least a nucleophilic reagent in the group that is made of following material, and described material is a water; can be the azanol that 1～4 alkyl replaces by carbonatoms; ammonia; carbonatoms is 1～4 alkylamine; carbonatoms is 2～8 dialkylamine; can be selected from carbonatoms is 1～4 alkyl; carbonatoms is that 6～10 aryl and carbonatoms are the hydrazine that the substituting group in 2～7 the alkyloyl replaces; with by carbonatoms be the quaternary ammonium halides that 1～6 alkyl replaces; and their chemically acceptable salt.
9. the described method of claim 1, wherein, at least a alkyl acetate, carry out operation (c) to (2S, 3R, 4S)-4-hydroxyisoleucine or its chemically conversion of acceptable salt.
10. the described method of claim 9, wherein, at least a alkyl acetate is an ethyl acetate.
11. (2S, 3R, 4S)-4-hydroxyisoleucine or its chemically the separation of acceptable salt remove method, wherein, described method comprises following operation (a) and (b),
Operation (a) the: make (2S in the mixture, 3R, 4S)-aldehyde cpd or the reaction of its equivalents that 4-hydroxyisoleucine or its chemically acceptable salt and formula Q-CHO represent, be converted into the compound of formula (1) expression, in the formula, it is that 6～14 aryl, carbonatoms are that 1～10 alkyl, carbonatoms are cycloalkyl or 5 yuan of heterocyclic radical～10 yuan heterocyclic radicals of 3～10 that Q represents to have substituent carbonatoms
In the formula, Q as defined above;
Operation (b): the compound that with an organic solvent extracts formula (1) expression.
12. the described method of claim 11, wherein, the aldehyde cpd that in the reaction of operation (a), uses formula Q-CHO to represent.
13. claim 11 or 12 described methods, wherein, in the formula (1), Q is for can have substituent phenyl, cyclohexyl, naphthyl, pyridyl, thienyl, furyl or pyrryl.
14. the described method of claim 13, wherein, in the formula (1), Q is for can have substituent phenyl.
15. claim 13 or 14 described methods, wherein, this substituting group when Q has substituting group is selected from the group that is made of following radicals:
(i) carbonatoms be 1～6 alkyl,
(ii) carbonatoms be 1～6 alkoxyl group,
(iii) carbonatoms be 2～7 alkyloyl,
(iv) carbonatoms be 6～10 aryl,
(v) carbonatoms be 6～10 aryloxy,
(vi) halogeno-group,
(vii) carbonatoms be 1～6 haloalkyl,
(viii) nitro,
(ix) formyl radical and
(x) cyano group,
Wherein, above-mentioned (i) with (vii) be not the substituting group of 1～10 alkyl as carbonatoms.
16. the described method of claim 11, wherein, the organic solvent of operation (b) is at least a alkyl acetate.
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| JP2007042711 | 2007-02-22 | ||
| JP042711/2007 | 2007-02-22 | ||
| JP144578/2007 | 2007-05-31 |
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