CN1073946A

CN1073946A - The method for preparing borate proline

Info

Publication number: CN1073946A
Application number: CN92113215A
Authority: CN
Inventors: R·斯诺; T·A·凯利; J·亚丹母斯; S·科特斯; C·帕理
Original assignee: Boehringer Ingelheim Pharmaceuticals Inc
Current assignee: Boehringer Ingelheim Pharmaceuticals Inc
Priority date: 1991-11-22
Filing date: 1992-11-23
Publication date: 1993-07-07
Also published as: NO941905L; FI942345A0; PT101079A; NZ245207A; NO941905D0; SK59294A3; AU3136893A; JPH07501078A; HUT67937A; SI9200332A; WO1993010127A1; AU661362B2; TW232697B; HU9401350D0; MX9206628A; FI942345A; EP0641347A1; CZ124494A3; IL103817A0; CA2123128A1

Abstract

Introduced a kind of method for preparing borate proline.By lithiumation, its product and trialkyl borate react the protected pyrroles of N-in the 2-position.Again it is reduced to form protected proline(Pro) boric acid, make it then to react to obtain ester with glycol.Along with the boric acid part with the ester group protection is removed the protecting group on the nitrogen, with the borate proline that need to obtain.The borate proline that makes in interchangeable synthesizing has a chiral centre on the boron atom.The method that enantiomorph splits is also disclosed, to obtain having borate proline rather than a kind of amino acid whose peptide at the C-end group.These boric acid peptide analogs can be used for suppressing important protein enzyme biologically.Several methods of removing the pinane glycol from the pinane glycol borate ester are also disclosed.

Description

The method for preparing borate proline

The present invention relates to a kind of method for preparing the borate proline of optically-active.These esters can be used as the intermediate of producing peptide and combine with proline(Pro) boric acid rather than proline(Pro).These peptides can be used to suppress the various enzymes of important protein biologically.

Holding residue as the peptide with the boric acid analogue that mixes a-amino acid to substitute C-, has been reports of effective inhibitor of many serine proteases based on these compounds, makes people produce interest to the boric acid analogue of a-amino acid.For example, referring to people such as Matteson [J.Am.Chem.Soc.103,5241(1981)]; People such as Kettner [J.Biol.Chem.259,15106(1984)]; With people [J.Med.Chem.27,1919(1985)] such as Kinder.

Some investigators, comprise people such as Koehler [Biochemistry, 10,2477(1971)] and people [Biochemistry such as Rawn, 13,3124(1974)], infer that the P track of the sky of the former subcenter of boron acts on mutually with the zymophore hydroxyl in these compounds, formed the tetrahedron adducts of the transition state of an analog enzyme hydrolysis.Can imagine that the boric acid analogue is more tight to the binding ratio substrate of enzyme itself, thereby suppress enzymatic activity substrate.Because the boric acid analogue of a-amino acid is luminous based on the biological function of the enzyme that they suppressed with the Toplink of mixing these analogues, so just be used at present in the research.In addition, explain that they also can be used in the treatment just as following.

Because being presented at the peptide that the C-end group is combined with the alpha-amino boronic acid analogue (BoroPro) of proline(Pro) is effective inhibitor of some postproline cleavage enzyme, be interested especially therefore.For example, people such as Bachovchin [J.Biol.Chem.265,3738(1990)] report the inhibitor that this class peptide is the IgA proteolytic enzyme that obtains from some bacterium.These enzymes are participated in bacterium toxicity consumingly.People such as Flentke [Proc.Natl.Acad.Sci.USA88,1556(1991)] report this class peptide and suppress dipeptidyl peptidase IV (DP-IV), and the latter then plays restraining effect to the cell proliferation and the IL-2 generation of antigen induction in the T-cell.Known back one effect causes immunoreactive inhibition.Immunoreactive being suppressed at handled as being useful in organ-graft refection, graft versus host disease and the various autoimmune disease.

The synthetic route of alpha-amino boronic acid in the past depends on the disclosed steps of people such as aforesaid Mattesont, and it follows hydroboration, carries out the response hierarchy that (asymmetric) homologation and ammonia are separated with the chloromethyl lithium.People such as Matteson [Organometallics 3,1284(1984)] have described and have utilized this technology to synthesize N-ethanoyl L-Ala, N-ethanoyl Xie Ansuan, the method for the boric acid analogue of N-ethanoyl leucine and N-acetyl phenyl alanine.Some are arranged in these analogues, and (9: 1) non-mapping foreign body object ratio obtains with preferably.

Utilize the synthetic BoroPro of method of Matteson to be confirmed by aforesaid Bachovachin and Flentke, need to make up pyrrolidine ring, therefore can only abandon but further modify.In addition, the reaction conditions that utilizes asymmetric synthesis or disassemble technique to prepare the BoroPro of single enantiomer yet there are no report.

Owing to the effective way that lacks proline biosynthesis boric acid has hindered further exploration postproline cleavage enzyme, the particularly biological chemistry of DP-IV, and the effort of using BoroPro base enzyme inhibitors to treat.

Needs to the better source of BoroPro impel us to explore this compound, the alternative synthetic route of the optical activity form of this compound particularly, and finished the present invention.

First big aspect of the present invention comprises the method for three proline biosynthesis boric acid esters that are closely related.Wherein two are begun by the pyrroles, and the 3rd synthetic method begun by tetramethyleneimine.In the scope of a first aspect of the present invention, comprise the intermediate that some are new.Proline(Pro) boric acid has a α chiral centre on the boron atom.Second big aspect of the present invention comprises a kind of method that splits proline(Pro) boric acid enantiomorph when needed.According to the method, by with a chiral alcohol, especially preferably react to generate borate proline with the pinane glycol.Separate the non-enantiomer mixture of introducing additional chiral centre generation and can finish the fractionation of enantiomorph easily.The borate proline that obtains can be at an easy rate and those activatory, as carbonyl acid groups phase coupling commonly used in peptide is synthetic, and obtaining having borate proline at the C-end group, rather than amino acid whose peptide.Can remove the ester protecting group to obtain free boric acid peptide.When the ester protecting group is the pinane glycol, be not easy to remove it with known technology.The 3rd aspect of the present invention comprises several methods of removing pinane glycerol protection base.

Fig. 1 is the preferred reaction scheme example of the present invention.

First synthetic method that begins with the pyrroles according to the present invention is to react with a kind of activatory carbonic acid derivatives, protects nitrogen-atoms so that utilize suc as formula the group of-COOR, and R is C in the formula _1-6Alkyl, C _3-6Cycloalkyl, benzyl, phenyl, by one or more C _1-6The phenyl that alkyl replaces, or trimethyl silyl ethyl are to obtain the compound suc as formula I.

R is the tertiary butyl, benzyl, trimethyl silyl ethyl, phenyl, methyl or ethyl in the better protecting group.Best protecting group is tertbutyloxycarbonyl or Boc.Can utilize known technology to use blocking group.People such as Grehn [Angew.Chem.Int.Ed.Engl.23.296, (1984)] have described the synthetic of 1-Boc-pyrroles.

Handle formula I compound to obtain compound with lithiation reagent subsequently suc as formula II

R is as previously defined group in the formula.

The lithiumation of formula I compound can be handled with known method with the methyl piperidine lithium and finish, as people such as Hasan [J.Org.Chem.46.157(1981)] introduce, or finish with other steric hindrance lithium amide such as diisopropylaminoethyl lithium or dicyclohexyl lithium amide, also can when having Tetramethyl Ethylene Diamine to exist, carry out lithiation with n-Butyl Lithium.This reaction can be carried out in inert solvent easily, is preferably temperature ether such as THF at-78 ℃～-40 ℃, diethyl ether, the solvent that glycol dimethyl ether or methyl tertiary butyl ether are such.Also available known as people such as Chen [Org.Syn.70.151(1991) method described with pyrroles's bromination, and protects resulting product on the 2-position, select for use as n-Butyl Lithium so not really the lithiation reagent of costliness to carry out lithiumation with known technology then.

Need not separate can be with the intermediate and the trialkyl borate reaction of formula II; wherein each alkyl can be the straight chain that contains 1～6 carbon atom; side chain or cycloalkyl; be preferably boric acid trimethylammonium ester or triethyl ester; carry out acid-catalyzed hydrolysis with the weak acid as citric acid or acetate or sal enixum so then, to obtain the pyrroles who has protecting group-2-boric acid suc as formula III

R is defined as the front in the formula.

Through catalytic hydrogenation formula III intermediate is reduced then, to form the proline(Pro) boric acid of protected formula IV.

The B definition is ditto described in the formula.

Formula III intermediate can use in the organic solvent as ethyl acetate or tetrahydrofuran (THF) as 5% platinum on the gac, platinum oxide, rhodium on the gac, the rhodium on the alumina, the such catalyzer of palladium on the gac or Raney Ni are under atmospheric pressure or carry out catalytic hydrogenation under the pressure of about 50psi.

A selectable synthetic method of the boric acid of formula IV is to handle tetramethyleneimine with a kind of suitable acylating reagent, to obtain protected compound suc as formula VIII.

R is defined as the front in the formula.

Select protecting group so that the activity of tetramethyleneimine can be carried out lithiumation in the position of contiguous nitrogen, protecting group should contain can hinder the huge part that lithiation reagent is attacked carbonyl.Preferably use the formamyl protecting group suc as formula-COOR, for example, R is tert.-butoxy or 2,4 in the group, 6-three-tertiary butyl phenoxy group.But also can use some acyl group or benzoyl group, as tertiary butyl carbonyl or trityl carbonyl.People such as Beak [Chem.Rev.84,471-523, (1984)] have described other suitable protecting group.Best protecting group is among tert-butoxycarbonyl or the Boc.Can protecting group be used for tetramethyleneimine by known technology.

Handle formula VIII compound to obtain formula IX compound with lithiation reagent subsequently.

R is defined as the front in the formula.

Formula VIII compound can be handled by known method with s-butyl lithium and Tetramethyl Ethylene Diamine and carry out lithiumation (described as people such as Beak [Tet.Lett.30.1197(1989)]).This reaction can be preferably a kind of ether such as diethyl ether easily in a kind of inert organic solvents, among methyl tertiary butyl ether or the THF, be approximately-78 ℃～0 ℃ in temperature, is preferably under-78 ℃～-40 ℃ the condition to carry out.Carry out lithiation with a reactive lithium alkylide such as s-butyl lithium or tert-butyl lithium, be preferably exist a kind of as Tetramethyl Ethylene Diamine, hexamethylphosphoramide or N, the N '-such coordination additive of dimethyl allene urea (DMPU).

Need not separate, formula IX intermediate then reacts with trialkyl borate, and wherein each alkyl can be the straight chain that contains 1～6 carbon atom, and side chain or cycloalkyl are preferably boric acid trimethylammonium ester or triethyl ester; Water is hydrolyzed and is extracted in the alkali aqueous solution as sodium hydroxide or potassium hydroxide to help purifying subsequently.Acidizing alkali solution is to about pH3, and extraction obtains the protected proline(Pro) boric acid suc as formula IV.

In order to form boric acid ester, the free boric acid intermediate that then makes the formula IV carries out esterification with glycol suc as formula V

X is a linking group in the formula, to obtain the compound suc as formula VI

X is and identical connection base above-mentioned in the formula, and R is defined as the front.So the ester group that forms is a kind of removable protecting group.The structure of this ester protecting group, synthetic be connected and the method for removing is generally known in chemical field.Therefore, the professional in the chemical field can be appreciated that the structure that connects basic X is unimportant.Connecting basic X can be 2 to 3 yuan of saturated hydrocarbon chains; Constitute and optionally contain unsaturated chain or condensed ring C _5-122 to 3 yuan of saturated hydrocarbon chains of the part of carbocyclic ring system; Constitute 2 to 3 yuan of hydrocarbon chains of the part of aromatic ring; Or suc as formula-(CH ₂) _n-NH-(CH ₂) _m-group, wherein n and m all 2 or 3; Wherein these groups can be unsubstituted or by one or more C _1-3Alkyl or phenyl replaces.

Correspondingly, be suc as formula the suitable glycol of V, for example, ethylene glycol, quite any alcohol, pyrocatechol, pinane glycol, fourth-2,3-glycol, 2,2-dimethyl propylene-1,3-glycol, diethanolamine and 1,2-phenylbenzene second-1,2-glycol.

For the boric acid part by ester group protection, available then known technology is removed protecting group on the denitrification (as Greene at " Protective Groups in Organic Synthesis " (J.Wiley; Sons, 1981) described in); To obtain hydrochloride suc as formula the required borate proline of VII.

For example, when protecting group was Boc, the dried hydrogenchloride that can be used at an easy rate in the ethyl acetate was removed.

Be preferably formula IV compound and a kind of chirality, non-racemic glycol are carried out esterification, these glycol are as (1S, 2S, 3R, 5S)-(+) pinane glycol), 1,2-phenylbenzene second-1,2-glycol; Or fourth-2, the 3-glycol; Because can in molecule, introduce additional chiral centre like this.Can utilize known diastereomer separation method like this, as the α chiral centre on HPLC or the fractionation crystallization fractionation boron atom.This point has illustrated in the reaction scheme figure of Fig. 1, in the isomers of formula VI compound with (1S, 2S, 3R, 5S)-(+)-boric acid of pinane glycerol protection can separate with high pressure liquid chromatography (HPLC) method (HPLC), obtains compound VI a and VI b.In addition; hydrochloride isomer with formula VII compound of same boron protecting group can be at solvent (as ethyl acetate; or dichloromethane/ethyl acetate mixture; Virahol or ethanol) in separate with fractionation crystallization, to obtain being connected carbon on the boron individual isomer compound VII b of R configuration is arranged.

Another benefit of using the pinane glycol is that the boric acid ester that so forms is more stable than the ester that obtains with other glycol, for example, often observes in chromatography when quite which is pure and has lost a large amount of protecting groups.Owing to can obtain the better rate of recovery of required material, this method is of great use carrying out in purifying and the isomer separation with silica gel column chromatography.

In the above-mentioned synthetic method that makes improvements slightly, the pyrrole derivative of formula III can be directly carries out esterification with the glycol of formula V, obtains the ester of formula III A.

This ester can reduce with the method identical with formula III compound, produces the protected borate proline suc as formula VI.In other words, promptly can put upside down pyrrole ring in reactions steps is reduced and the esterified order of boronate.

Zhi Bei borate proline is easy to and activatory carboxylic acid commonly used in peptide is synthetic like this, and for example, the protected amino acid coupling of a kind of nitrogen is to obtain the compound suc as formula X

In the formula-COOR ²Be a kind of amino protecting group commonly used in peptide is synthetic, so R ²Be, for example, the tertiary butyl, benzyl or fluorenyl methyl, R ³Be naturally occurring amino acid whose side chain, can have the sort of suitable protecting group commonly used in peptide is synthetic.

Formula X compound contains protecting group on boric acid and amino acid nitrogen.In order to keep biologic activity or, one of them or two protecting groups can be removed in case of necessity for further chemical treatment.Can remove protecting group by arbitrary order.The whole bag of tricks of removing these protecting groups is described below.

Available known method is removed nitrogen-protecting group, to obtain the compound of formula XI.

The ester group of protection boric acid part is stable in neutral or acid organic medium, but the cracking and produce boric acid rapidly under alkali (pH=7.5) aqueous conditions of gentleness of many boric acid esters.Yet, under the boric acid ester situation of using the pinane glycol, known its hydrolysis is difficult, and need specific conditions to remove the pinane glycol, for example, referring to people [J.Organometallic Chem.385,15(1988)] such as people such as Matteson [J.Am.Chem.Soc.102,7590(1980)] and Brown.

These methods are unsuitable for removing the pinane glycol in the compound of X or XI from suc as formula VI.We have found that some are used for removing the method for pinane glycol in the boric acid ester compound of X or XI from suc as formula VI.Therefore, the removal of pinane glycol can use a kind of oxygenant with cracking 1,2 glycol ability to carry out under mild conditions, to remove the pinane glycol and balance is further moved to free boric acid direction from balance.For example, handling protecting group with sodium metaperiodate under room temperature in ammonium acetate solution and acetone is the compound of the formula VI and the formula X of pinane glycol, obtains the compound of formula IV and formula XII respectively.

This reaction can be carried out in water easily, can add the buffered soln as ammonium acetate or Sodium phosphate dibasic, pH is between 3～10, be preferably 6 to 8, temperature is 0 to 80 ℃, be preferably 20 to 40 ℃, can have a kind of as acetone, methyl alcohol, ethanol, tetrahydrofuran (THF) (THF) or acetonitrile etc. can be miscible with water organic solvent.Suitable oxygenant is to have to decompose 1, the non-nucleophilicity oxygenant of 2-glycol ability such as Periodic acid or its salt or permanganate.Do not find the oxygenolysis of carbon one boron key under these conditions, be appreciated that this method is applicable in any boric acid with the pinane glycerol protection.In addition, it also can be used for anyly belonging to 1, and in the boric acid ester protecting group of 2-glycol, and when to the simple aqueous hydrolytic action of protecting group when very slow or incomplete, this method is particularly useful.

Contain the pinane glycol borate ester compound of the amine of not protection in use, during suc as formula the XI compound, also can use aforesaid method, but more desirable to second kind of novel method of such compound.This method be included in pH4 or the cation exchange resin column of formula XI compound water solution being packed into when lower in, water or this post of dilute acetic acid wash-out are to remove the pinane glycol.Can from balance, remove the pinane glycol like this and reaction is moved to the hydrolysis direction.Then with rare this post of ammonium hydroxide aqueous solution wash-out removing product, obtain the salt of formula X III compound after evaporation and the acidifying

Be to use a kind of strong-acid cation-exchange resin aptly, as sulfonic resin, Dowex50 for example, the pinane glycol of wash-out can reclaim from the aqueous solution and reuse from resin, this can pass through a nonionic superpolymer sorbent material with the aqueous solution easily, as can almost quantitatively adsorbing the Amberlite of pinane glycol

The XAD-200 post is finished.The pinane glycol can be removed from post with methyl alcohol or ethanol elution.Ion-exchange and pinane glycol adsorb this two steps operation and can be attached in the process, with pump water are recycled to another pillar from a pillar in this process.Have the still less advantage of water gaging like this, and can make process continuously to sufficiently long to obtain higher product yield.

One of skill in the art can understand that the above-mentioned method of utilizing a kind of Zeo-karb to remove the pinane glycol only is suitable for containing the alkaline functional base, as the compound of unprotected amine.

The third method also is suitable for containing the compound of not protecting amine, suc as formula the pinane diol ester of XI compound.This method is included in the two-phase system and uses formula R ⁴-B(OH) ₂Another boric acid carry out the transesterification of pinane glycol borate ester, R in the formula ⁴Represent the alkyl of a C1-12, it can be by straight chain, and side chain or cycloalkyl chain and phenyl ring are formed, preferred R ⁴It is phenyl.In the two-phase one is that pH adjusts to and is lower than 7 mutually, is preferably the water of pH1-4, and another is as hexane mutually, the hydrocarbon organic solvent that sherwood oil or toluene are such.Therefore, in the water of pH1 and hexanes mixtures, handle formula XI compound, by being separated, in organic phase, produce the pinane diol ester of phenyl-boron dihydroxide subsequently, it can be reclaimed through evaporation with phenyl-boron dihydroxide; And at the solution of the free boric acid of aqueous phase production X III, spent ion exchange resin with above-mentioned similar methods can be with this solution separating.The unique component that is dissolved in this system in the organic phase is a pinane glycol phenyl boronate, thereby has removed the pinane glycol from balance; The formula X III compound of formula XI compound and formation is because the two all is insoluble to varsol, so they are stayed in the water layer.Available any boric acid that has the alkyl side chain reacts, and generates the pinane diol ester that is dissolved in the varsol.

The also available known method of complete de-protected formula X III compound is removed nitrogen-protecting group and is prepared from formula XII compound.As can be seen suc as formula X, XI, the compound of XII and X III has two chiral centres usually.A contiguous boron atom, another is present in the amino acid moiety, when this part makes an exception during for glycine.Also can see,, compare the pure single diastereomer that more needs these compounds with the non-enantiomer mixture of these compounds for biological applications.Therefore it is very important to prepare the pure individual isomer of these compounds.The most important thing is that because amino acid may exist with the single enantiomer form usually, this purpose can be with known technology by making the coupling and separate formed non-enantiomer mixture and reach mutually of optically pure amino acid and racemize proline(Pro) boric acid.Yet, have been found that these Special Circumstances of Xie Ansuan only, this class separate often be difficulty with time taking.Therefore better being to use a kind of chiral centre that is close to the boron atom is the proline(Pro) boric acid of individual isomer, at this moment with after the coupling of optically-active pure amino acid need not carry out the fractionation of isomer.The invention provides a kind of method that is easy to be used to split proline(Pro) boric acid enantiomorph.

The following example further illustrates the present invention.

Embodiment 1

1-(1,1-dimethyl ethoxy carbonyl)-pyrroles-2-boric acid

To tetrahydrofuran (THF) (THF) (275ml) under-78 ℃ of argon shields tetramethyl piperidine (8.8ml, 52mmol) add in the solution 2M solution of butyllithium in ethane (26ml, 52mmol).In above-mentioned solution, be added in THF(10ml after 15 minutes) in 1-(1,1-dimethyl ethoxy carbonyl)-pyrroles (8.35g, 50mmol) solution, and stirred 4 hours down at-78 ℃.(30ml 176mmol), and made mixture rise to room temperature through 3 hours, again after 12 hours, with ether (500ml) diluted reaction mixture and use 1M KHSO to add triethyl borate then ₄The aqueous solution wash (3 * 100ml), use 1M NaHCO then ₃The aqueous solution wash (1 * 100ml), use MgSO ₄Drying gets brown solid through rotary evaporation, and it is used flash chromatography method purifying (1: 9 ethyl acetate: hexane), obtain 8.7g(82%) white crystalline solid (101.0～101.5 ℃ of fusing points) on silica gel.

¹H NMR(CDCl ₃): δ 1.65(s, 9H), and 6.26(t, J=3.3(Hz, 1H), and 7.10(dd, J=1.6,3.2Hz, 1H), 7.15(s, 2H), and 7.44(dd, J=1.6,3.2Hz, 1H); 13C NMR(CDCl ₃) 27.9,85.5,112.0,128.7,152.0; CIMS m/z(% relative intensity) 212(MH+, 11), 156(100), 138(68); Ultimate analysis: C ₉H ₁₄BNO ₄Calculated value: C, 51.23, H, 6.69, N, 6.64. measured value: C, 51.22, H, 6.51, N, 6.67.

Embodiment 2

1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2-boric acid

6.15 gram (24mmol) is as 1-(1 of embodiment 1 preparation, 1-dimethyl ethoxy carbonyl)-the solution 5%pt/c(ca.500mg of pyrroles-2-boric acid in ethyl acetate (100ml)) hydrogenation 24 to 48 hours under 50psi.The suspension of gained filters and concentrates through Celite pad.This material carries out chromatography with silica gel, and used continuous elutriant is 9: 1 hexanes: ethyl acetate is acetone then, and the concentrated trace solvent post crystallization of removing of partial acetone is gone out limpid glassy required compound 6.05 gram (97%) (100～101 ℃ of fusing points).

¹H NMR(CDCl ₃): δ 1.42(s, 9H), 1.6-2.15(m, 5H), 3.1-3.6(m, 2H); ¹³C NMR(CDCl ₃) δ 25.1,25.7,28.4,45.6,46.2,78.6,154.5; CIMS m/z(% relative intensity) 116(100), 70(46); Ultimate analysis: C ₉H ₁₈BNO ₄Calculated value: C, 50.27, H, 8.44, N, 6.51.

Measured value: C, 50.52, H, 8.22, N, 6.58.

Embodiment 3

(1S, 2S, 3R, 5S)-pinane glycol 1-(1,1-dimethyl ethoxy carbonyl-tetramethyleneimine-2S-boric acid ester and (1S, 2S, 3R, 5S)-pinane glycol 1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2R-boric acid ester

Press the 1-(1 of embodiment 2 preparations, 1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2-boric acid (1.52g, 7.1mmol) solution and (1S, 2S, 3R, 5S)-(+)-(1.36g 8.0mmol) stirred under the room temperature 2 hours in ether (25ml) the pinane glycol, concentrated and through flash chromatography on silica gel (85: 15 hexanes: ethyl acetate) obtain the mixture of two kinds of diastereomers of 2.1 grams (85%) 1: 1 ratio.They carry out HPLC through the porous silica gel A post of a 300 * 3.9mm and separate, and use methyl tertiary butyl ether: hexane (1: 9) wash-out and usefulness U.V detector are in the detection of 220nm place.Under these conditions at first wash-out be isomer (1S, 2S, 3R, 4S, 5S)-pinane glycol 1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2S-boric acid ester.

The S-isomer: ¹H NMR(C ₆D ₆): δ 0.55(s, 3H), 1.09(s, 3H), 1.52(s, 9H), 1.60(s, 3H), 1.2-2.2(m, 8H), 3.1-3.5(m, 3H), 4.11(m, 0.3H), 4.33(m, 0.7H); ¹³C NMR(C ₆D ₆): δ 23.9,26.6,27.1,27.3,28.4,28.6,28.8,36.0,38.2,39.9,46.1,51.9,78.3,78.5,85.7,154.9.

The R-isomer: ¹H NMR(C ₆D ₆): δ 0.52(s, 3H), 1.08(s, 3H), 1.52(s, 9H), 1.61(s, 3H), 1.2-2.2(m, 8H), 3.1-3.6(m, 3H), 4.01(m, 0.3H), 4.25(m, 0.7H); ¹³C NMR(C6D6) 23.9,26.6,27.1,27.3,28.4,28.7,28.9,35.8,38.2,39.6,46.2,51.8,78.1,78.5,85.7,154.5.

Can clearly be seen that, with (1R, 2R, 3S, 5R)-(-)-the pinane glycol makes raw material and can prepare (1R, 2R, 3S with similar method, 5R)-pinane glycol-1(1, the 1-dimethyl ethoxycarbonyl)-tetramethyleneimine-2S-boric acid ester and (1R, 2R, 3S, 5R)-and pinane glycol-1-(1, the 1-dimethyl ethoxycarbonyl)-tetramethyleneimine-2R-boric acid ester.

Embodiment 14

(1S, 2S, 3R, 5S)-pinane glycol tetramethyleneimine-2S-boric acid ester hydrochloride

Press (1S, 2S, 3R, 5S)-pinane glycol-1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2S-acid ester solution (28.5mg, the 0.08mmol) stirring in the EtoAc of anhydrous HCl solution (about 3M) of embodiment 3 preparation.This solution concentrates 2 times to produce 21.2 milligrams (91%) required white salt acidifying thing solid (molten 204 ℃ (decomposition)) after 2 hours from ethyl acetate.

¹H NMR(CDCl ₃): δ 0.83(s, 3H), and 1.14(d, J=11Hz, 1H), 1.29(s, 3H), 1.45(s, 3H), 1.85-2.15(m, 6H), 2.17-2.50(m, 3H), 3.18-3.25(m, 1H), 3.45(bs, 2H), 4.42(dd, J=1.8,8.6Hz, 1H), 8.80(bs, 1H), 10.56(bs, 1H); ¹³C NMR(CDC ₁₃): δ 23.9,24.5, and 26.5,27.0,27.2,28.4,34.9,38.2,39.4,45.8,51.2,79.0,87.6; CIMS m/z(% relative intensity) 250(MH+, 1000); HRMS(EI): C ₁₄H ₂₄BNO ₂Calculated value 249.1900, measured value 249.1899.

As can be seen with similar method can prepare (1S, 2S, 3R, 5S)-pinane glycol-tetramethyleneimine-2S-boric acid ester hydrochloride.

Embodiment 5

(1S, 2S, 3R, 5S)-pinane glycol tetramethyleneimine-2R-boric acid ester hydrochloride

Press (the 1S of embodiment 3 preparations, 2S, 3R, 5S)-pinane glycol-1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2R-acid ester solution (18.3mg, 0.05mmol) handle with the anhydrous HCl among the aforesaid EtoAc, back acquisition 14.3mg(96% is finished in experiment) required white salt acidifying thing solid (fusing point 248 ℃ (decomposition)).

¹H NMR(CDCl ₃): δ 0.83(s, 3H), and 1.14(d, J=11Hz, 1H), 1.29(s, 3H), 1.45(s, 3H), 1.85-2.15(m, 6H), 2.17-2.50(m, 3H), 3.18-3.25(m, 1H), 3.45(bs, 2H), 4.42(dd, J=1.8,8.6Hz, 1H), 8.80(bs, 1H), 10.56(bs, 1H); ¹³C NMR(CDCl ₃) δ 23.9,24.5,26.5,27.0,27.2,28.5,34.9,38.1,39.4,45.8,51.2,79.0,87.8; CIMS m/z(% relative intensity) 250(MH+, 100); HRMS(EI): C ₁₄H ₂₄BNO ₂Calculated value 249.1900, measured value 249.1903.

As can be seen with similar method can prepare (1R, 2R, 3S, 5R)-pinane glycol-tetramethyleneimine-2R-boric acid ester hydrochloride.

Embodiment 6

(1S, 2S, 3R, 5S)-and pinane glycol-1-(1,1-dimethyl ethoxy carbonyl)-pyrroles-2-boric acid ester

Press the 1-(1 of embodiment 1 preparation, 1-dimethyl ethoxy carbonyl)-pyrroles-2-boric acid solution (1.36g, 6.45mmol) and (1S, 2S, 3R, 5S)-(+)-(1.10g 6.45mmol) stirred 4 hours in the 20ml ether pinane glycol.Rotary evaporation is flash chromatography (95: 5 hexanes: ethyl acetate), obtain 1.83 gram (82%) required limpid oily products on silica gel then.

¹H NMR(CDCl ₃): δ 0.90(s, 3H), 1.30(s, 3H), 1.41(d, J=11Hz, 1H), 1.50(s, 3H), 1.50(s, 3H), 1.59(s, 9H), 1.96(m, 2H), and 2.21(t, J=6Hz, 1H), 2.16-2.40(m, 2H), 4.45(dd, J=2,8Hz, 1H), 6.20(t, J=3Hz, 1H), 6.65(d, J=3Hz, 1H), 7.40(d, J=3Hz, 1H); ¹³C NMR(CDCl ₃): δ 24.0,26.4, and 27.2,28.1,28.6,35.5,38.3,39.8,51.8,79.8,83.6,83.9,111.6,123.2,124.7,150.0; CIMS m/z(% relative intensity) 346(MH+, 4), 246(81), 153(100), 135(56).

Can prepare with similar method as can be seen (1R, 2R, 3S, 5R)-and pinane glycol-1(1,1-dimethyl ethoxy carbonyl)-pyrroles-2-boric acid ester.

Embodiment 7

From 1-(1,1-dimethyl ethoxycarbonyl)-tetramethyleneimine prepares 1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2-boric acid

To the 1-(1 under-78 ℃ of nitrogen protections; 1-dimethyl ethoxy carbonyl)-tetramethyleneimine (17.1g; 100mmol) add in the solution in Anaesthetie Ether (200ml) solution of 1.3M s-butyl lithium in hexanaphthene (92.3ml, 120mmol) in, keep temperature to be lower than-60 ℃ simultaneously.After adding reaction mixture was stirred 4 hours down at-78 ℃.(31.1g 300mmol), made mixture temperature rise to room temperature through 3 hours to add boric acid trimethylammonium ester.After 12 hours, add entry (150ml) again, add 2M NaOH(200ml subsequently).Water phase separated is also used 2M NaOH(150ml) extracted organic phase once more.(5 * 200ml) extract the alkaline extraction liquid that merges with 2M HCl acidifying to pH3 and with ethyl acetate.With the organic extract drying (Na that merges ₂SO ₄) and concentrate to form required white crystalline solid product (15.49g, 72%), the material that itself and embodiment 2 produce is identical.

Embodiment 8

(1S, 2S, 3R, 5S)-and pinane glycol-1-(1, the 1-dimethyl ethoxycarbonyl)-tetramethyleneimine-2RS-boric acid ester

To the material that makes by embodiment 7 (15.49g, 72.0mmol) add in the solution in chloroform (250ml) (1S, 2S, 3R, 5S)-(+)-the pinane glycol (12.77g, 7.5mmol).After stirring 16 hours under the room temperature nitrogen protection, remove and desolvate; with flash chromatography on silica gel method (hexane/ethyl acetate 9: 1; 4: 1) the purifying residuum; obtain required oily non-enantiomer mixture (23.6g in 1: 1 ratio; 67.7%; press 1-(1,1-dimethyl ethoxy carbonyl)-the tetramethyleneimine meter).The isomer mixture that this material and embodiment 3 make is identical.

¹H NMR(CDCl ₃): δ 0.85(s, 3H), 1.12-1.21(m, 1H), 1.29(s, 3H), 1.41(s, 3H), 1.45(s, 9H), 1.81-2.20(m, 8H), 2.28-2.39(m, 1H), 3.04-3.18(m, 1H), 3.34-3.45(m, 2H), 4.28-4.38(m, 1H); ¹³C NMR(CDCl ₃): δ 23.7,26.2, and 27.1,28.5,35.5,38.2,39.6,46.1,78.0,78.8,85.7,85.8,154.7; CIMS m/z(% relative intensity) 350(MH+, 100), 294(72), 250(30).

Embodiment 9

(1S, 2S, 3R, 5S)-analytical procedure of pinane glycol-tetramethyleneimine-2-boric acid ester hydrochloride diastereomer

The reagent solution of preparation 0.2M thiocarbanil in methylene dichloride-triethylamine (9: 1).Analyzed sample (1-5mg) is handled with reagent solution by the amount of every micromole's assay 10 microlitre reagent solutions, and clear liquid was placed 15 minutes in room temperature.Then, 1 microlitre solution example is analyzed (post: YMCAQ-303 S-5 120A, 4.6 * 250mm with 1.00 milliliters of high pressure liquid chromatography (HPLC) level dilution in acetonitrile, the solution after 10 microlitres dilute like this with the high pressure liquid chromatography (HPLC) method, moving phase: 65%MeCN-35% 25mM ammonium phosphate, pH7.5; Flow velocity 1ml/ branch detects at the 254nm place with UV).Wash-out goes out the phenylthiourea derivative of proline(Pro) boric acid R isomer in the time of about 6.4 minutes, and wash-out goes out its epimer in the time of about 7.8 minutes, as the unreacted thiocarbanil of internal standard then 12.2 minutes the time wash-out come out.

Embodiment 10

(1S, 2S, 3R, 5S)-pinane glycol tetramethyleneimine-2RS-boric acid ester hydrochloride

(1S, 2S, 3R with ice-cooled stirring, 5S)-pinane glycol-1-(1,1-dimethyl ethoxy carbonyl)-(224g, the 0.64mmol) solution in diethyl ether (900ml) fed solution 35 minutes with anhydrous HCl gas to tetramethyleneimine-2RS-boric acid ester under 10～18 ℃.Solution at room temperature stirs and spends the night, once more with ice-cooled and filter out throw out.Solid is washed then with sherwood oil/diethyl ether 9 with cold diethyl ether (400ml): 1(200ml) wash and vacuum-drying, obtain (fusing point 228-234 ℃ of required white salt acidifying thing solid (113g, 62%).Analyze this material with the high pressure liquid chromatography (HPLC) method described in the embodiment 9, prove that it is a R:S mixture of isomers by the boric acid of 60: 40 mixed.

¹H NMR(CDCl ₃): δ 0.83(s, 3H), and 1.14(d, J=11Hz, 1H), 1.29(s, 3H), 1.45(s, 3H), 1.85-2.15(m, 6H), 2.17-2.50(m, 3H), 3.18-3.25(m, 1H), 3.45(bs, 2H), 4.42(dd, J=1.8,8.6Hz, 1H), 8.80(b, 1H), 10.56(bs, 1H); ¹³C NMR(CDCl ₃) δ 23.9,24.5,26.5,27.0,27.2,28.4,34.9,38.2,39.4,45.8,51.2,79.0,87.6; CIMS m/z(% relative intensity) 250(MH+, 100); Ultimate analysis: C ₁₄H ₂₄BNO ₂HCl calculated value: C, 58.87, H, 8.82, N, 4.90, Cl, 12.41. measured value: C, 58.40, H, 8.86, N, 4.81, Cl, 12.39.

Hydrochloride (about 4.5M with ether, 200ml) and ethyl acetate (EtoAc) (150ml) make solvent, carry out 29g, the similar reaction of 101mmol scale can obtain the R by 81: 19 mixed: the mixture of S isomer hydrochloride (11.1g, 47%).

Embodiment 11

Utilize that fractionation crystallization obtains (1S, 2S, 3R, 5S)-pinane glycol-tetramethyleneimine-2R-boric acid ester hydrochloride

Method A:

(1.18g 4.13mmol) is dissolved in CH to 60: 40 isomer mixtures that embodiment 10 is obtained ₂Cl ₂(65ml), heat a little and solution is filtered.Filtrate is diluted with ethyl acetate (65ml), and crystallization begins within one minute.Suspension at room temperature stirred 1-2 hour, collected the solids that forms for the first time, by recording diastereomer ratio (540mg, 46%, R: S ratio 97.1: 2.9) described in the embodiment 9.Solvent is distilled up to most of CH from filtrate ₂Cl ₂Be removed, then remaining ethyl acetate solution at room temperature stirred the micro white solids (346mg, 29%, the R: S ratio 39.2: 60.8) that spend the night and form for the second time to produce.Primary product Virahol (10ml) recrystallization acquisition 430mg(80% transformation efficiency) the 2-R isomer material (fusing point 269-272 ℃ (decomposition)) of purity＞99%.〔α〕 ₂₅D+0.70°（C＝1.15，MeOH）

¹H NMR(CDCl ₃): δ 0.83(s, 3H), and 1.14(d, J=11Hz, 1H), 1.29(s, 3H), 1.45(s, 3H), 1.85-2.15(m, 6H), 2.17-2.50(m, 3H), 3.18-3.25(m, 1H), 3.45(bs, 2H), 4.42(dd, J=1.8,8.6Hz, 1H), 8.80(bs, 1H), 10.56(bs, 1H); ¹³C NMR(CDCl ₃) δ 23.9,24.5,26.5,27.0,27.2,28.5,34.9,38.1,39.4,45.8,51.2,79.0,87.8, CIMS m/z(% relative intensity) 250(MH+, 100); Anal.Calcd for C ₁₄H ₂₄BNO ₂HCl: C, 58.87, H, 8.82, N, 4.90, Cl, 12.41. measured value: C, 58.64, H, 8.79, N, 4.90, Cl, 12.66.

Method B:

(1S, 2S, 3R, 5S)-isomer mixture (850mg of pinane glycol-tetramethyleneimine-1: 1 ratio of 2RS-borate acidulants, 2.98mmol) suspension in ethyl acetate (60ml) stirred 4 hours under reflux, with the mixture heat filtering and collect the material (541mg, 64%) that the exsiccant solids obtains enrichment R isomer, wherein R: the S ratio is 2: 1.Filtrate evaporation is obtained the material (217mg) of enrichment S isomer, wherein R: the S ratio is 1: 4.The solids that obtains after filtration (500mg) was handled 1.5 hours with ethyl acetate (45ml) with same method, carried out heat filtering again to obtain solids (366mg, 73%), R: S=7: 1.This material uses ethyl acetate (38ml) to handle once more 1.5 hours, and the R of separated solids this moment (287mg, 78%): the S ratio is 97: 3.Its spectral quality is identical with the material that is obtained by the A method.

Embodiment 12

Prepare 1-(1,1-dimethyl ethoxy carbonyl from (1S, 2S, 3R, 5S)-pinane glycol 1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2-boric acid ester)-tetramethyleneimine-2-boric acid

To (the 1S that makes by embodiment 8 described methods, 2S, 3R, 5S)-pinane glycol-1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2RS-boric acid ester (1.9g, 5.44mmol) add in the solution in acetone (80ml) 0.1M ammonium acetate solution (80ml) and sodium metaperiodate (3.49g, 16.33mmol).Reaction mixture is stirring at room 40 hours, and then with acetone evaporated, residuum is with 2M NaOH solution-treated, this water CH ₂Cl ₂(2 * 80ml) extractions.With the organic extract liquid drying (Na that merges ₂SO ₄) and concentrate to produce needed white foam shape solid product (890mg, 76%), its NMR with the material of preparation in embodiment 2 is identical.For analysis purposes is derived boric acid and tetramethyl ethylene ketone.

Embodiment 13

Any pure 1-(1 quite, 1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2RS-boric acid ester

Boric acid (the 890mg that in embodiment 12, produces, 4.14mmol) add quite any alcohol (489mg in the solution in chloroform, 4.14mmol), at room temperature stir to remove after 16 hours and desolvate, residuum is by silica gel column chromatography (hexane/ethyl acetate, 4: 1) carry out purifying, obtain needed white solid product (1.04g, 85%) (fusing point: 73-75 ℃).

¹H NMR(CDCl ₃): δ 1.18(s, 6H), 1.21(s, 6H), 1.38(s, 9H), 1.57-2.00(m, 4H), 2.98(br s, 1H), 3.27(m, 2H); 13C NMR(CDCl ₃): δ 24.3,24.5, and 24.7,24.9,25.3,27.0,27.6,28.4,28.6,43.6,45.8,46.3,78.8,83.2,154.4,154.8; CIMS m/z(% relative intensity) 298(18), 242(100, MH+-tBu), 198(30, MH+-Boc); Ultimate analysis: C ₁₅H ₂₈BNO ₄Calculated value: C, 60.62, H, 9.50, N, 4.71. measured value: C, 60.94, H, 9.65, N, 4.88.

Embodiment 14

N-(1,1-dimethyl ethoxy carbonyl)-the valyl tetramethyleneimine of L--2R-boric acid (1S, 2S, 3R, 5S)-the pinane diol ester.

(351.7g is 1.62mol) at CH for tert-butoxycarbonyl (t-Boc)-L-Xie Ansuan ₂Cl ₂Solution (1.6L) cools off with ice bath, and 0～2 ℃ within 40 minutes with dicyclohexyl carbodiimide (161.8g, CH 0.784mol) ₂Cl ₂(0.75L) solution adds in the above-mentioned solution.After adding solution was stirred 3.5 hours down at 0～5 ℃, filter out white precipitate and use CH ₂Cl ₂(0.2L) wash, in the settled solution that obtains, add (1S, the 2S for preparing by embodiment 10 18～20 ℃ (water-bath coolings), 3R, 5S)-(210g 0.735mol) is containing N-methylmorpholine (164g, CH 1.62mol) to pinane glycol tetramethyleneimine-2RS-boric acid ester hydrochloride ₂Cl ₂Solution (2.0L).Mixture is in stirred overnight at room temperature, and muddy solution filters through a high silicagel column (200～425 order) of 16cm diameter * 2cm, and uses CH ₂Cl ₂(1.5L) wash.Solvent evaporated is to obtain the oily matter (542g) of high viscosity.This mixture will be cooled off in this oily matter dissolving dry ethyl acetate (0.7L) and in ice bath.The crystallization that filters out formation is at low temperatures also washed with cold ethyl acetate (0.1L).Wet cake transferred in the sherwood oil (0.65L) stirred 1 hour under the room temperature.The white solid thing is filtered, wash and be dried to constant weight to obtain the subject compound (113.4g) (128～130 ℃ of fusing points) of white solid with cold sherwood oil (0.1L).Collect all mother liquors and be concentrated into about 0.8 liter of volume.In refrigerator, place 2 days after-filtration and go out formed solids, handle to obtain beige solid (50.4g) with sherwood oil as stated above.This solids is the mixture of impurity and unwanted diastereomer.Concentrate the mother liquor that obtains above, residuum is (14cm diameter * 60cm on silicagel column; With hexane/ethyl acetate (85: 15) (14L) open up the layer) purifying it.Collect suitable part and handle, collect solids and dry to obtain more required product (18.5g) with filtration method with sherwood oil.Obtain another kind of diastereomer (7.5g) (fusing point 82-83 ℃) simultaneously.Mixing portion that merges and mother liquor obtain extra pure compound (13.5g) by second column purification, obtain 145.4g(44.3% altogether) required diastereomer (128～130 ℃ of fusing points).

¹H NMR(CDCl ₃): δ 0.83(s, 3H), and 0.91(d, J=6.7Hz, 3H), 0.97(d, J=6.7Hz, 3H), 1.27(s, 3H), 1.35-1.45(m, 1H), 1.39(s, 3H), 1.41(s, 9H), 1.72-2.14(m, 9H), 2.26-2.36(m, 1H), 3.15(dd, J=6.7,10.1Hz, 1H), 3.43-3.51(m, 1H), 3.70-3.81(m, 1H), 4.19-4.28(m, 2H), 5.29(d, J=9.2Hz, 1H); ¹³C NMR(CDCl ₃): δ 17.3,19.2, and 24.0,26.3,27.1,27.2,27.4,28.4,28.6,31.4,33.9,35.5,38.2,39.6,46.7,51.2,56.6,77.8,79.2,85.8,155.9,170.2; CIMS m/z(% relative intensity) 449(MH+, 100), 393(50); Ultimate analysis: C ₂₄H ₄₁BN ₂O ₅Calculated value: C, 64.28, H, 9.22, N, 6.25. measured value: C, 64.58, H, 9.33, N, 6.52.

Embodiment 15

L-Val-Pyr-2R-boric acid (1S, 2S, 3R, 5S)-pinane diol ester acid maleate

With N-(1; 1-dimethyl ethoxy carbonyl)-L-Val-Pyr-2R-boric acid (1S; 2S; 3R; 5S)-pinane diol ester (248mg; 0.553mmol) join in the solution of anhydrous hydrogen chloride in ethyl acetate of stirring, solvent evaporated is to stay de-protected hydrochloride after 1.5 hours.Residuum is allocated in CH ₂Cl ₂And between the sodium carbonate solution, organic layer is dry on sal epsom, and organic layer contains the free alkali of subject compound, and it exists with the ring compound form that contains a nitrogen-boron key, is the open loop form but restore after adding acid.Organic solution is filled into toxilic acid, and (64mg, 0.553mmol) in the solution in methyl alcohol (5ml), solvent evaporated stays crystalline residuum (258mg), and its recrystallization in ethyl acetate is obtained subject compound (193mg, 75%)

(145～146 ℃ of fusing points).

¹H NMR(CDCl ₃): δ 0.84(s, 3H), and 1.08(d, J=6.9Hz, 3H), 1.13(d, J=6.9Hz, 3H), 1.26-1.31(m, 2H), 1.29(s, 3H), 1.38(s, 3H), 1.72-2.15(m, 7H), 2.24-2.38(m, 2H), 3.28(dd, J=6.9,9.4Hz, 1H), 3.38-3.47(m, 1H), 3.73-3.78(m, 1H), 4.14(d, J=5.1Hz, 1H), 4.26(d, J=7.1Hz, 1H), 6.25(s, 2H), 7.5-9.0(v.br, 4H); ¹³C NMR(CDCl ₃): δ 17.0,18.4, and 24.0,26.3,27.0,27.1,28.7,30.0,35.4,38.2,39.5,47.3,51.2,56.6,78.1,86.2,135.6,166.3,169.5; CIMS m/z(% relative intensity) 349(MH+, 100), 197(18); Ultimate analysis: C ₂₃H ₃₇BN ₂O ₇Calculated value: C, 59.49, H, 8.03, N, 6.03. measured value: C, 59.50, H, 8.13, N, 6.03.

Embodiment 16

L-Val-Pyr-2R-boric acid mesylate

A) ring-L-Val-Pyr-2R-boric acid

(5.0g, 10.8mmol) (1.0%, 60ml) solution in adds and is H the maleate that will obtain by embodiment 15 at dilute acetic acid ^-On the Dowex50X2-200 ion exchange resin column of form (3.5cm dark * 4cm diameter), use then acetic acid (1.0%, 14L), water (42L) and solution of ammonium hydroxide (by commodity 0.880 solution that dilutes at 1: 100) this post of wash-out.The pinane glycol can reclaim from center pillar and acidic moiety.Product appears at basic moiety early, with its collection and use CH ₂Cl ₂(2 * 100ml) wash.With the dry free alkali (174mg, 20%) that also concentrates the parent material that obtains reclaiming with some pinane glycol of organic extract.To form subject compound, there be (1.52g, 66%) (120～130 ℃ of fusing points) in this compound with the ring-type white solid compound form with a nitrogen-boron key with the water lyophilize.

¹H NMR(CDCl ₃): δ 0.97(d, J=7.0Hz, 3H), 1.06(d, J=7.0Hz, 3H), 1.59-1.80(m, 2H), 1.95-2.03(m, 2H), 2.41-2.51(m, 1H), 2.62-2.69(m, 1H), 3.23-3.32(m, 1H), 3.51-3.58(m have the eclipsed doublet, J=4.2Hz, 2H); ¹³C NMR(D ₂O): δ 19.0,21.7, and 27.3,30.7,29.9,49.6,61.0,170.3; IR(cm ^-1) 3400-3314,3221-3108,2961-2872,1637,1452-1369; CIMS m/z(% relative intensity) 375(90, M ₂H+-3H ₂O), 197(100, MH+-H ₂O); Ultimate analysis: C ₉H ₁₉BN ₂O ₃Calculated value: C, 50.50, H, 8.95, N, 13.09. measured value: C, 50.43, H, 8.76, N, 12.93.

B) L-Val-Pyr-2R-boric acid mesylate

Dropwise to the cyclisation boric acid (5.17g that as above makes under nitrogen protection that stirs; 24.16mmol) add methylsulfonic acid (2.32g in the suspension in acetonitrile (190ml); 24.16mmol) solution in acetonitrile (10ml); after dripping in 5 minutes; mixture was at room temperature stirred 2 hours; collect solids with filtration method, with acetonitrile and diethyl ether that its thorough washing is also dry to obtain to be white in color solid subject compound (6.14g, 82%) (179～180 ℃ of fusing points).This material through dimethyl formamide/acetonitrile crystallization once obtains this material (fusing point 181-182 ℃) of 70% yield.

¹H NMR(D ₂O, phosphoric acid salt, pH2): δ 0.99(d, J=6.8Hz, 3H), 1.09(d, J=6.9Hz, 3H), 1.69-1.75(m, 1H), 1.90-1.99(m, 1H), 2.10-2.14(m, 2H), 2.28-2.35(m, 1H), 2.80(s, 3H), 3.07(dd, J=7.0 and 11.2Hz, 1H), 3.46-3.51(m, 1H), 3.75(t, J=9.0Hz, 1H), 4.14(d, J=5.1Hz, 1H); At 3.53-3.55(m) and 3.83(d, J=6.2Hz) locate also to find cis acid amides rotational isomer; ¹³C NMR: δ 16.2,18.4,26.9,27.1, and 29.0,38.8,47.9,49.0,57.2,167.2; 16.8,24.3,29.9,57.8,167.5 places find to belong to the peak of cis acid amides rotational isomer; IR(cm ^-1) 3387,3000(br), 2972,2655,1646,1370,1197; CIMS m/z(% relative intensity, eg adduct bond) 241(MH+100);

Ultimate analysis: C ₁₀H ₂₃BN ₂O ₆S

Calculated value: C, 38.72, H, 7.47, N, 9.03.

Measured value: C, 38.65, H, 7.45, N, 8.44.

Embodiment 17

N-(1,1-dimethyl ethoxy carbonyl)-L-Val-Pyr-2R-boric acid

To the N-(1 that presses embodiment 14 preparations that stirs, 1-dimethyl ethoxy carbonyl)-L-Val-Pyr-2R-boric acid (1S, 2S, 3R, 5S)-(1.0g 2.3mmol) adds ammonium acetate solution (60ml to the pinane diol ester in the solution in acetone (75ml), 0.1M) and sodium metaperiodate (1.48g, 6.91mmol), reaction mixture was stirring at room 48 hours, and acetone is removed in evaporation then.CH is handled and used to residuum with 2M sodium hydroxide solution (100ml) ₂Cl ₂Wash (2 * 50ml).Water layer is acidified to pH3 carefully and uses CH with 2M hydrochloric acid ₂Cl ₂(4 * 70ml) extractions.The acidic solution organic extract liquid that merges also concentrates to form required white foam shape solid product (700mg, 90%) with dried over sodium sulfate.Through silica gel column chromatography (CH ₂Cl ₂/ methyl alcohol 9: 1) be further purified and obtain white boric acid solids (449mg, 62%) (fusing point: 82～92 ℃) once more.

¹H NMR(CDCl ₃): δ 0.95(d, J=5.7Hz, 6H), 1.42(s, 9H), 1.55-1.80(m, 1H) 1.80-2.20(m, 4H), 2.89-3.07(m, 1H), 3.30-3.55(m, 1H), 3.55-3.65(m, 1H), 4.10-4.30(m, 1H), and 5.34(d, J=9.2Hz, 1H); ¹³C NMR(CDCl ₃): δ 18.0,19.1, and 26.3,27.7,28.3,31.2,46.1,52.0,55.7,79.5,155.6,170.8; IR(cm ^-1) 3395-3319,2971-2875,1711,1619,1400,1174; CIMS m/z(% relative intensity, eg adduct bond)

341（MH ⁺，100），285（MH ⁺-tBu，67），241（MH ⁺-BOC，21）.

Embodiment 18

The valyl tetramethyleneimine of L--2R-borate acidulants

The N-(1 that makes by embodiment 17; 1-dimethyl ethoxy carbonyl)-the valyl tetramethyleneimine of L--2R-boric acid (250mg; 0.796mmol); under the room temperature nitrogen protection with HCl/ ether (4.5M; 20ml) stir 1.5 hours together; solvent is removed in evaporation then, and (3 * 10ml) grind and the ether that at every turn inclines residuum with diethyl ether.With the residuum drying to obtain the solid-state subject compound of white powdery (172mg, 86%) (211～213 ℃ of fusing points).

¹H NMR(D ₂O, phosphoric acid salt pH2): δ 0.99(d, J=6.9Hz, 3H), 1.09(d, J=7.0Hz, 3H), 1.67-1.76(m, 1H), 1.87-2.01(m, 1H), 2.09-2.15(m, 2H), 2.28-2.35(m, 1H), 3.07(dd, J=7.0 and 11.4 Hz, 1H), 3.48(dt, J=6.7 and 10.3Hz, 1H), 3.73(dt, J=1.7 and 10.2Hz, 1H), 4.14(d, J=5.2Hz, 1H); ¹³C NMR: δ 16.0,18.3,26.9,27.1, and 28.9,47.9,48.9,57.2,167.3; IR(cm ^-1) 3400-2800,3368,2970/2880,1635,1475-1378,1400; CIMS m/z(% relative intensity, eg adduct bond) 241(MH ⁺, 100).

Embodiment 19

Carry out the valyl tetramethyleneimine of transesterification preparation ring-L--2R-boric acid with phenyl-boron dihydroxide

Will be by the valyl tetramethyleneimine of the L--2R-boric acid (1S of embodiment 15 preparations; 2S; 3R; 5S)-pinane diol ester hydrochloride (500mg; 1.3mmol) (500mg, the solution that is become in the 1M hydrochloric acid (10ml) 2.6mmol) be violent stirring 1 hour at room temperature containing hexane (20ml) and phenyl-boron dihydroxide.Remove hexane with decantation, add more hexane (20ml) then, and mixture was further stirred 30 minutes.Divide each layer of leaving away, the hexane layer of merging is with dried over sodium sulfate and concentrated pinane glycol phenyl-borate (331mg, 99%) with the crystalline solid that obtains being white in color.Water layer is by a Dowex50 ion exchange resin column subsequently, and this post water (20ml) wash-out is used solution of ammonium hydroxide (commodity 0.880 solution of dilution in 1: 100,50ml) wash-out subsequently.Separate basic moiety, lyophilize obtains the free boric acid (230mg, 83%) of white powdered then, and it is identical to analyze the material that itself and embodiment 16a obtain through NMR.

Claims

1, a kind of proline(Pro) of preparation formula VII

The method of boric acid ester

X is a linking group in the formula,

This method comprises:

A) handle the pyrroles with the activatory carbonic acid derivatives, to obtain suc as formula the protected pyrroles of the N-of I

R is C in the formula _1-6Alkyl, C _3-6Cycloalkyl, benzyl, phenyl is by one or more C _1-6The phenyl that alkyl replaces, or trimethyl silyl ethyl,

B) handle the protected intermediates of formula I with lithiation reagent, to obtain the intermediate of formula II

R is as defined above in the formula,

C) with trialkyl borate and the reaction of formula II intermediate, each alkyl can be the straight chain that contains 1-6 carbon atom in this ester, and side chain or cyclic alkyl are used acid-catalyzed hydrolysis then, to obtain the intermediate of formula III

R is as defined above in the formula,

D) with catalytic hydrogenation reduction-type III compound, to obtain the proline(Pro) intermediate of formula IV

R is as defined above in the formula,

E) make formula IV compound with suc as formula the reaction of the glycol of V

X is and identical connection above-mentioned in the formula

Base is to obtain the boric acid ester of formula VI

X is and identical connection base above-mentioned in the formula, and R is as defined above,

F) remove the protecting group on the nitrogen-atoms in the proline(Pro) ring.

2, according to the process of claim 1 wherein in (C) step Chinese style II intermediate and boric acid trimethylammonium ester or the reaction of triethyl ester.

3, according to the process of claim 1 wherein that connecting basic X is 2 to 3 yuan of saturated hydrocarbon chains; Constitute C _5-122 to 3 yuan of saturated hydrocarbon chains of the part of carbocyclic ring system, this member ring systems can contain unsaturated link(age) or condensed ring; Constitute 2 to 3 yuan of hydrocarbon chains of the part of aromatic ring system; Or X is formula-(CH ₂) n-NH-(CH ₂) group of m-, wherein n and m all can be 2 or 3; Wherein this class group can be unsubstituted or by one or more C _1-3Alkyl or phenyl replaces.

4, according to the process of claim 1 wherein that quite where esterification in (e) step is with the glycol of selecting oneself, alcohol, pyrocatechol, pinane glycol, fourth-2, the 3-glycol, diethanolamine and 1,2-phenylbenzene second-1, the glycol of 2-glycol is finished.

5, according to the method for claim 4, wherein glycol is the pinane glycol of optically active.

6, according to the method for claim 5, the protected pinane diol ester of N-that wherein forms in (e) of claim 1 step further is separated into its diastereomer.

7, according to the method for claim 5, the protected pinane diol ester of N-that wherein forms in (f) of claim 1 step further is separated into its diastereomer.

8, according to the process of claim 1 wherein that R is the tertiary butyl, benzyl, trimethyl silyl ethyl, phenyl, methyl or ethyl.

9, a kind of method of borate proline of preparation formula VII,

X connects base in the formula,

This method comprises:

A) handle the pyrroles with a kind of activatory carbonic acid derivatives, with the protected pyrroles of N-of acquisition formula I,

R is C in the formula _1-6Alkyl, C _3-6Cycloalkyl, benzyl, phenyl is by one or more C _1-6

The phenyl that alkyl replaces, or trimethyl silyl ethyl,

B) handle formula I protected intermediates with lithiation reagent, obtaining the intermediate of formula II,

R is as defined above in the formula,

C) with trialkyl borate and the reaction of formula II intermediate, each alkyl can be the straight chain that contains 1～6 carbon atom in this ester, and side chain or cyclic alkyl are used acid-catalyzed hydrolysis subsequently, to obtain the intermediate of formula III

R is as defined above in the formula,

D) glycol of formula V and formula III compound are reacted

V is to connect base in the formula,

To obtain the boric acid ester of formula III A

R is as defined above in the formula,

E) ester that is generated through the catalytic hydrogenation reduction is to obtain the proline(Pro) intermediate of formula VI

F) remove protecting group from the proline(Pro) ring nitrogen.

10, according to the method for claim 9, wherein in (c) step, formula II intermediate and boric acid trimethylammonium ester or triethyl ester react.

11, according to the method for claim 9, wherein connecting basic X is 2 to 3 yuan of saturated hydrocarbon chains; Constitute C _5-122 to 3 yuan of saturated hydrocarbon chains of the part of carbocyclic ring system, this member ring systems can contain unsaturated link(age) or condensed ring; Constitute 2 to 3 yuan of hydrocarbon chains of the part of aromatic ring system; Or X is formula-(CH ₂) n-NH-(CH ₂) group of m-, wherein n and m each naturally 2 or 3; Wherein this class group can be unsubstituted or by one or more C _1-3Alkyl or phenyl replaces.

12, according to the method for claim 9, wherein the esterification in (e) step is to comprise ethylene glycol, quite which alcohol, pyrocatechol, pinane glycol, fourth-2 with being selected from, 3-glycol, diethanolamine and 1, and 2-phenylbenzene second-1, the glycol of 2-glycol is finished.

13, according to the method for claim 12, glycol wherein is the pinane glycol of optically active.

14, according to the method for claim 13, the protected pinane diol ester of N that wherein forms in (e) of claim 9 step further is separated into its diastereomer.

15, according to the method for claim 13, the protected pinane diester of N that wherein forms in (f) of claim 9 step further is separated into its diastereomer.

16, according to the method for claim 9, wherein R is the tertiary butyl, benzyl, trimethyl silyl ethyl, phenyl, methyl or ethyl.

17, a kind of method of preparation formula IV intermediate

This method comprises:

A) handle the pyrroles with the activatory carbonic acid derivatives, with the protected pyrroles of the N-that obtains the formula I,

B) handle the protected intermediates of formula I with lithiation reagent, obtaining formula II intermediate,

R is as defined above in the formula,

C) with the intermediate reaction of trialkyl borate and formula II, each alkyl can be straight chain, side chain or the cycloalkyl that contains 1-6 carbon atom in this ester, uses acid-catalyzed hydrolysis subsequently, with the intermediate of acquisition formula III,

R is as defined above in the formula,

D) through the compound of catalytic hydrogenation reduction-type III, to obtain the proline(Pro) intermediate of formula IV.

18, a kind of method of intermediate of preparation formula IV

This method comprises:

A) handle tetramethyleneimine with acylating reagent, with the protected tetramethyleneimine of acquisition formula VIII,

R is C in the formula _1-6Alkyl, C _3-6Cycloalkyl, benzyl, phenyl are by one or more C _1-6Phenyl or trimethyl silyl ethyl that alkyl replaces,

B) handle the compound of formula VIII with lithiation reagent, to obtain the compound of formula IX

R is as defined above in the formula,

C) with the compound reaction of trialkyl borate and formula IX, each alkyl contains straight chain, side chain or the cycloalkyl of 1-6 carbon atom in this ester,

D) product of hydrolysis abovementioned steps is to obtain the compound of formula IV.

19, according to the method for claim 18, wherein in (c) step Chinese style IX intermediate and boric acid trimethylammonium or the reaction of triethyl ester.

20, the intermediate of formula III

Wherein R is C _1-6Alkyl, C _3-6Cycloalkyl, benzyl, phenyl are by one or more C _1-6

The phenyl that alkyl replaces, or trimethyl silyl ethyl.

21,1-(1,1-dimethyl ethoxy carbonyl)-pyrroles-2-boric acid.

22, a kind of compound that is selected from down group comprises

(1S, 2S, 3R, 5S)-the pinane glycol

1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2S-boric acid ester;

(1S, 2S, 3R, 5S)-the pinane glycol

1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2R-boric acid ester; (1R, 2R, 3S, 5R)-the pinane glycol

1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2S-boric acid ester; (1R, 2R, 3S, 5R)-the pinane glycol

1-(1,1-dimethyl ethoxy carbonyl)-tetramethyleneimine-2R-boric acid ester.

23, a kind of compound that is selected from down group comprises

(1S, 2S, 3R, 5S)-pinane glycol tetramethyleneimine-2S-boric acid ester hydrochloride;

(1S, 2S, 3R, 5S)-pinane glycol tetramethyleneimine-2R-boric acid ester hydrochloride;

(1R, 2R, 3S, 5R)-pinane glycol tetramethyleneimine-2S-boric acid ester hydrochloride;

(1R, 2R, 3S, 5R)-pinane glycol tetramethyleneimine-2R-boric acid ester hydrochloride.

24, a kind of compound that is selected from down group comprises

(1S, 2S, 3R, 5S)-and pinane glycol 1-(1,1-dimethyl ethoxy carbonyl)-pyrroles-2-boric acid ester; With

(1R, 2R, 3S, 5R)-and pinane glycol 1-(1,1-dimethyl ethoxy carbonyl)-pyrroles-2-boric acid ester.

25, a kind of removing from boric acid ester in the method as the pinane glycol of protectiveness ester group, this method comprise with a kind of having cracking 1, this class boric acid ester of the oxidizer treatment of 2-glycol ability.

26, according to the method for claim 25, wherein said oxygenant is a sodium metaperiodate.

27, a kind of pinane glycol of from formula X compound, removing as the protectiveness ester group

R in the formula ²Be a blocking group, R ³Be naturally occurring amino acid whose side chain, it can have suitable protecting group; Or the method for from formula XI compound, removing the pinane glycol,

R in the formula ²Be as defined above, this method comprises with a kind of having cracking 1, this class boric acid ester of the oxidizer treatment of 2-glycol ability.

28, according to the method for claim 27, wherein said oxygenant is a sodium metaperiodate.

29, a kind of method of from boric acid ester, removing as the pinane glycol of protectiveness ester group with free amine; this method comprises that the aqueous solution with this class boric acid ester is added on the cation exchange resin column; water and this post of dilute acid solution wash-out to be to remove the pinane glycol, at last with this post of dilute alkaline aqueous solution wash-out to remove free boric acid product.

30, according to the method for claim 29, wherein said column packing is a kind of strongly acidic cation-exchange.

31, a kind of method of from the compound of formula XI, removing as the pinane glycol of protectiveness ester group

R in the formula ³It is naturally occurring amino acid whose side chain; it can have suitable protecting group; this method comprises this class boric acid ester aqueous solution is added on the cation exchange resin column; water and dilute acid solution wash-out to be to remove the pinane glycol, at last with this post of dilute alkaline aqueous solution wash-out to remove free boric acid product.

32, according to the method for claim 31, wherein said column packing is a kind of strongly acidic cation-exchange.

33, the intermediate of formula X

Protecting group-O-X-O derives from the pinane glycol in the formula, and R wherein ²Be protecting group, R ³Be naturally occurring amino acid whose side chain, it optionally has suitable protecting group.

34, N-(1,1-dimethyl ethoxy carbonyl)-the valyl tetramethyleneimine of L--2R-boric acid (1S, 2S, 3R, 5S)-the pinane diol ester.

35, the intermediate of formula XI

Protecting group-O-X-O is derived from the pinane glycol in the formula, and R wherein ³Be the natural amino acid whose side chain that exists, it optionally has suitable protecting group.

36, the valyl tetramethyleneimine of L--2R-boric acid (1S, 2S, 3R, 5S)-pinane diol ester or its salt.

37, a kind of method of borate proline of preparation formula VII

X connects base in the formula, and this method comprises:

A) handle tetramethyleneimine with acylating reagent, to obtain the protected tetramethyleneimine of formula VIII

Wherein R is C _1-6Alkyl, C _3-6Cycloalkyl, benzyl, phenyl are by one or more C _1-6The phenyl that alkyl replaces, or trimethyl silyl ethyl,

R such as preceding definition in the formula,

C) compound of trialkyl borate and formula IX is reacted, each alkyl all can be straight chain, side chain or the cycloalkyl that contains 1～6 carbon atom in this ester

D) product in the hydrolysis abovementioned steps is to obtain the compound of formula IV

E) with the glycol of formula V and the compound reaction of formula IV

X is the connection base identical with top described person in the formula, to obtain the boric acid ester of formula VI

X carries identical connection base with top in the formula, and R is as defined above,

F) remove the protecting group on the nitrogen-atoms in the proline(Pro) ring.

38, according to the method for claim 37, wherein in intermediate and the boric acid trimethylammonium ester or the reaction of triethyl ester of (C) step Chinese style IX.

39, according to the method for claim 37, wherein linking group X is 2 to 3 yuan of saturated hydrocarbon chains; Constitute C _5-122 to 3 yuan of saturated hydrocarbon chains of the part of carbocyclic ring system, this member ring systems alternative has unsaturated link(age) or condensed ring; Constitute 2 to 3 yuan of hydrocarbon chains of the part of aromatic ring system, or X is formula-(CH ₂) n-NH-(CH ₂) group of m-, wherein n and m all 2 or 3; Wherein this class group can be unsubstituted or by one or more C _1-3Alkyl or phenyl replaces.

40, according to the method for claim 37, wherein the esterification in (e) step is to comprise ethylene glycol with being selected from, any alcohol quite, and pyrocatechol, the pinane glycol, fourth-2, the 3-glycol, diethanolamine and 1,2-phenylbenzene second-1, the glycol of 2-glycol is finished.

41, according to the method for claim 40, glycol wherein is the pinane glycol of optically active.

42, according to the method for claim 41, the protected pinane diol ester of N that wherein forms in (e) of claim 37 step further is separated into its diastereomer.

43, according to the method for claim 41, the protected pinane diol ester of N that wherein forms in (f) of claim 37 step further is separated into its diastereomer.

44, according to the method for claim 37, wherein R is the tertiary butyl, benzyl, trimethyl silyl ethyl, phenyl, methyl or ethyl.