CN87105898A - N-(sProcess and intermediates for the preparation of -3-alkylheptanoyl -D-r-glutamyl -glycyl -D-alanyl - Google Patents

Abstract

Immunomodulator N-, (s-3-alkylheptanoyl) -D-r-glutamyl-glycyl-D-alanine and esters thereof; and novel immunomodulators N- (R-3-alkyl-4-heptenoyl-andsimproved synthesis processes and intermediates for -3-methyl-6-heptenoyl) -D-r-glutamyl-glycyl-D-alanine and esters thereof.

Description

The present invention relates to the favourable preparation method of the immunomodulator shown in the formula I,

R is methyl or ethyl in the formula I; And R ⁴And R ⁵Respectively be hydrogen, or R ⁴And R ⁵Among one be hydrogen, another is (C ₁-C ₆) alkyl or (C ₆-C ₈) methyl cycloalkyl; The invention still further relates to the preparation method and the intermediate of trans-R-3-alkyl-4-heptenoic acid (VI b sees below), S-3-methyl-6-heptenoic acid (X b sees below) and S-3-alkyl-enanthic acid, the absolute stereo chemical formula of S-3-alkyl enanthic acid is

R stipulates as above in the formula II, and they can be used as the intermediate of synthetic formula I compound; The present invention relates to the absolute stereo chemical formula shown in (III) immunomodulator (or precursor) or its can be used as medicine salt,

On behalf of two keys rather than both, any in the formula III in two dotted lines all represent pair keys simultaneously, and R stipulates as above, R ²And R ³The both be hydrogen (III a) or its can make pharmaceutical salt or R ²And R ³Be (C independently of one another ₁-C ₆) alkyl, (C ₆-C ₈) methyl cycloalkyl or benzyl (III b), collateral condition is when terminal 6, when the 7-position was two key, R was a methyl.

Optically pure S-3-methyl enanthic acid (II, R=CH ₃) originally obtained productive rate unspecified people J.Biol.Chem. such as (, 1932 95 volume 1-24 pages or leaves on 18 pages, are called 2-normal-butyl butyric acid-4 in the document) Levene by its quinine salt of recrystallization repeatedly at-15 ℃ by its corresponding raceme.Optically active 3-methyl enanthic acid is reported more available other method preparation (people such as Soai, J.Chem.Soc., Chem.Commun, 469-470 page or leaf in 1985 in succession; People such as Oppolzer, Helv.Chim.Acta.1985 68 volume 212-215 pages or leaves; People such as Ohno, United States Patent (USP) 1986 4,564,620; People such as Mori, Synthesis nineteen eighty-two 752-753 page or leaf; People such as Oppolzer, Helv.Chim.Acta.1981 64 volume 2808-2811 pages or leaves; People such as Mukaiyama, Chem.Lett.1981 913-916 page or leaf; People such as Posner, J.Am.Chem.Soc.1981 103 volume 2886-2888 pages or leaves; People such as Mukaiyama, Bull.Chem.Soc.Japan, 1978 51 volume 3368-3372 pages or leaves; People such as Meyers, J.Am.Chem.Soc.1976 98 volume 2290-2294 pages or leaves), (the maximum possible yield is 50%, has at least 50% unwanted by product to need to handle but these preparation methods have one or more shortcomings; Have at least 50% unwanted by product to take final several steps of operating process to always; The product acid that obtains is not optically pure; Need to use the organometallic reagent that is difficult to operate in the mass preparation; Total recovery is low; And/or required reagent is difficult to obtain).

(Katsuhi and Sharpless, J.Am.Chem.Soc., 1980 102 volume 5974-5976 pages or leaves were reported in now used asymmetric Epoxidation effect (so-called Sharples (Sharpless) fractionation) in the past; People such as Sharpless, Pure Appl.Chem., nineteen eighty-threes 55 are rolled up the 589-604 page or leaf); Also has stereospecific claisen (Claisen) condensation reaction (people such as Chan, United States Patent (USP) 4,045,475) in addition.

Newer immunopharmacology field particularly relates to the immunomodulatory part, obtains to develop rapidly.The compound of all natural generations is studied, comprises the appropriate Ford of tetrapeptide west (tuffsin), chemistry N-(1-(N-L-Threonyl-L-lysyl) by name-L-prolyl)-the L-arginine.Many handles are concentrated on synthetic glycan peptide derivant, and particularly those are called the compound of Muramyl dipeptide.

The immunomodulator of formula I (is worked as R ⁴=R ⁵Be generally amorphous lyophilize thing during=H) narrate among portable relevant therewith international monopoly (PCT) the application serial no PCT/US85/02351 that goes out to apply for as far back as November 25 in 1985 with their using method.Because people can't obtain this application publicly, the preparation of these compounds and their using method have been incorporated the disclosure into to support application.

Other immune excited peptide class is existing narration in some patent specifications:

L-alanyl-α-pentanedioic acid N-acyl group dipeptide, (seeing the Deutsches Reichs-Patent of announcing on January 15th, 1,981 3,024,355);

Contain four of D-alanyl-L-glutamyl part or L-alanyl-D-glutamyl part-and five-peptide class (seeing English Patent 2,053,231 and the Deutsches Reichs-Patent 3,024,281 announced on February 4th, 1981 respectively);

N-acyl group-alanyl-r-o-glutamyl tripeptide derivative, wherein the C-end amino acid is Methionin or diaminopimelic acid (seeing the Deutsches Reichs-Patent of announcing on January 15th, 1,981 3,024,369);

Comprise N-lactoyl alanyl, glutamyl, the lactoyl tetrapeptide class of diamino heptanedioyl and carboxymethylamino component (seeing the Europe patent EP-11283 that announced on May 23rd, 1980);

Polypeptide and derivative thereof with formula (A) are disclosed in United States Patent (USP) 4,311,640 and 4,322,341; European patent application 25,482, in 50,856,51,812,53,388,55,846 and 57,419,

R in the formula (A) ^aBe hydrogen or acyl group; R ^bAt first being hydrogen, also can be low alkyl group, methylol, benzyl; R ^cAnd R ^dRespectively be hydrogen, carboxyl, CONR ^gR ^h(R wherein ^gBe hydrogen, with the low alkyl group that hydroxyl has selection to replace, R ^hBe mono carboxylic, the dicarboxyl low alkyl group); R ^eBe hydrogen or carboxyl, collateral condition is to work as R ^dAnd R ^eAmong one be hydrogen, another then be carboxyl or-CONR ^gR ^h; R ^fBe hydrogen; M be 1～3 and n be 0～2, carboxyl in polypeptide and the derivative thereof and amino are shielded; And

Similar in appearance to the peptide class of following formula (A), but R wherein ^dForm amino acid moiety (people such as IV es, United States Patent (USP) 4,565,653 of an alkalescence; Or a heterocyclic amino acid (IV es, european patent application 178,845) european patent application 157,572).

People such as Kitaura are in J.Med.Chem.1982 25 volume 335-337 page or leaf reports, N ²(r-D-glutamyl)-meso-2(L), 2(D)-diaminopimelic acid is the minimal structure that (A) formula compound can be drawn the biologically characteristic, (A) n is 1 in the formula; M is 2; R ^aBe CH ₃CH(OH)-CO-; R ^bBe CH ₃; R ^cAnd R ^eRespectively be-COOH; R ^dBe-CONHCH ₂COOH; And R ^fBe H.Should be called FK-156 by (A) formula compound.

We have now found that a kind of immunomodulator of more stable crystal form, i.e. R=CH in the above-mentioned formula (I)₃And R⁴＝R ⁵The compound N of=hydrogen-(S-methyl heptanoyl group)-D-r-glutamyl-glycyl-D-alanine.

We have now found the high intermediate R-3-alkyl of preparation polarimetry purity-4-heptenoic acid (VI b sees below) and S-3-alkyl enanthic acid (II is seen before) effective method, its total recovery height, and avoid Use the synthetic middle useless accessory substance of R-enantiomer of organometallic reagent and later stage.

Article one, route, a step of beginning be make following formula trans-4-alkane alkene-3-alcohol carries out the Sharples type and splits,

The regulation of R as above in the formula. Specifically, this step comprises that the raceme that makes formula (IV) is having titanium tetraisopropylate and L-(+)-reaction-inert solvent that the tartaric acid diisopropyl ester exists in, with tert-butyl hydroperoxide reaction, the latter's amount should be enough to oxidation S-enantiomer and keep the R-enantiomer of required nullvalent absolute stereo chemical formula (V).

In second step, formula V compound stereotaxis ground and ortho-acetic acid three ((C₁-C ₃) alkyl) ester condensation, without separation, in the reactionlessness reagent that has acid to exist, intermediate pi-allyl-enol ether is reset, and obtains absolute stereo chemical formula (a) R-3-alkyl of VI-4-heptenoic acid (C₁-C ₃) Arrcostab,

(V 1a) X=OR ¹; R ¹=(C ₁-C ₃) alkyl

（Ⅴ1b）X＝OH

（Ⅴ1c）X＝Cl

（Ⅴ1d）X＝H

In another route, this ester then is hydrolyzed to trans-R-3-alkyl-4-heptenoic acid (V 1b) as needs, is hydrogenated to S-3-alkyl enanthic acid (II) again.Another can be selected in the route of usefulness, at first with this ester hydrogenation, gets S-3-alkyl enanthic acid (C ₁-C ₃) alkyl ester, be hydrolyzed to the acid shown in the formula II then.

In another route, the trans-4-alkane of fractionation is rare-and 3-alcohol (formula V) having catalyst acetic acid mercury (Hg(OAc) ₂) when existing, use the ethyl vinyl ether effect, be transformed into allyl vinyl ether (IX).Heating is reset to the R-3-alkyl-4-heptenal shown in the formula (V 1d) allyl vinyl ether then, reoxidizes, and for example uses Jones reagent (chromic anhydride in diluted mineral acid), forms above-mentioned R-3-alkyl-4-heptenoic acid.

We have also found intermediate S-3-methyl-6-heptenoic acid (X d sees below) and S-3-methyl enanthic acid (II, the R=CH for preparing high polarimetry purity ₃, see before) effective ways, the total recovery height avoids using organometallic reagent and useless R-enantiomorph by product, is raw material with the S-geraniol (XI) that the absolute stereo chemical formula is arranged that is easy to get.

It is conventional that this synthetic method starts several steps; comprise that with conventional silyl blocking group (as t-butyldimethylsilyl) protection of selection alcohol radical is arranged, then ozone decomposes, and handles with methyl-sulfide; obtain the new compound of absolute stereo chemical formula shown in (XII)

R in the formula (XII) ⁸Be that (XII a) or silyl hydroxyl protecting group (XII b) for hydrogen.Used method is special explanation in the preparation of back, and similar in appearance to give the method that is used for the R-geraniol earlier in the R-enantiomorph of preparation above-mentioned formula (V) compound, wherein R ⁸It is t-butyldimethylsilyl.The latter (the R-geraniol is of no use here) can be used for synthetic general former times good fortune and comes (proxipomin) people such as (, J.Chem.Soc., Chem.Commun.1985 143-145 page or leaf) Tapolczay.

The present invention be more particularly directed to formula (V) midbody compound and formula (X) compound,

(X is R a) ⁹=CH ₂OR ¹⁰, R ¹⁰=silyl protecting group

（Ⅹb）R ⁹＝CH ₂OH

（Ⅹc）R ⁹＝CHO

（Ⅹd）R ⁹＝COOH

（Ⅹe）R ⁹＝COCl

And relate to and change formula (XII) compound into optically active acid (formula (X d) and (II, R=CH ₃)) method, this method comprises the following steps:

(a) compound of formula (XII) and methylene tri Phenylphosphine CH ₂=P(C ₆H ₅) ₃Reaction, (X is a) or (X b) compound to form formula;

(b) when (X is a) when compound is formula (X b) compound, can be used as an independent step to carry out or carry out simultaneously with next step with the diluted mineral acid hydrolyzing type; With

(c) chromic anhydride oxidation-type (X b) compound that is used in the diluted mineral acid is S-3-methyl-6-heptenoic acid (formula X d); If desired, can once go on foot again,

(d) catalytic hydrogenation formula (X d) compound is that R is the S-3-methyl enanthic acid of methyl in the formula II.

R ⁸Be hydrogen, less in steps benefit, and R ⁸Be the silyl protecting group, then when by step (a) isolation of intermediate products, the benefit of easier purifying arranged.Preferred hydroxyl protecting group is the trimethyl-methane base, to tert.-butylbenzene ethyl dimetylsilyl and t-butyldimethylsilyl.Preferred diluted mineral acid is H ₂SO ₄Preferably select for use with S-geraniol ozone and decompose, (XII is compound a) with method synthesis type that methyl-sulfide is handled; And through following (e), (f) synthetic R of two steps ⁸It is formula (XII b) compound of t-butyldimethylsilyl.

(e) with S-geraniol and the reaction of chlorination t-butyldimethylsilyl; And

(f) geraniol that reacts the hydroxyl protection that obtains decomposes with ozone, handles with methyl-sulfide.

The present invention also relates to prepare the modification method of formula I immunomodulator, the method comprising the steps of:

(a) activated form (as acyl chlorides V 1c or X e) that makes R-3-alkyl-4-heptenoic acid or S-3-methyl-6-heptenoic acid and compound coupling in reaction-inert solvent shown in the formula (VII) form above-mentioned formula (III b) midbody compound,

In the formula (VII), R ⁶And R ⁷Both all are benzyls, or R ⁶And R ⁷Among one be benzyl, another is (C ₁-C ₆) alkyl or (C ₆-C ₈) methyl cycloalkyl.R in the formula (III b) ²And R ³Corresponding to the R in the formula (VII) ⁶And R ⁷; With

The reduction of two keys and the hydrogenolysis of one or more benzyls take place in (b) the above-mentioned midbody compound of hydrogenation in the reaction-inert solvent that has hydrogenation catalyst to exist simultaneously.

At last, the present invention relates to the precursor formula III of immunomodulator and/or above-claimed cpd (I).(III a) compound (is R to formula ²=R ³=hydrogen) be to obtain through the ester hydrolysis of routine by formula (III b) diester compound.

Here used " reaction-inert solvent " speech is meant such solvent, it with starting raw material, intermediate or product reaction, but it has adverse influence to the yield of required product.

Preferred R is an ethyl.Preferred acid catalyst be Lewis acid (be exsiccant HCl, AlCl ₃).The raceme of preferably trans-4-hexene-3-ol or trans-4-teracrylic acid-alcohol is to obtain with crotonic aldehyde or the reaction of 2-pentenals respectively by the ethyl Grignard reagent, but the present invention is not limited to by this.

The very easy realization of the present invention.R-is trans-and 4-alkane alkene-3-alcohol (V) is by the asymmetric Epoxidation effect, and promptly so-called Sharples Split Method (document of quoting previously) obtains.According to this method, in the raceme (IV) unwanted S-trans-2-hexene-4-alcohol is at the L-(+ that has more than 1 mole)-the tartrate diisopropyl ester be substantially in the presence of the titanium tetraisopropylate of 1 molar equivalent, optionally with tert-butyl hydroperoxide (at least 0.5 mole, but every mole of racemization hexenol certainly its consumption be less than 1 mole) reaction.This is reflected under the anhydrous condition and carries out in low temperature in reaction-inert solvent such as methylene dichloride, and general temperature range is-20～-80 ℃.Required unreacted R-is trans-and 2-alkane alkene-4-alcohol (V) separates with standard method (as distillation or chromatography).Because the volatility of this required product is higher, so the low boiling point solvent that is used for chromatographic separation is preferably as pentane and ether.

Next step be R-trans-4-alkane alkene-3-alcohol (V) and ortho-acetic acid (C ₁-C ₃) trialkyl ester condensation (claisen condensation rearrangement), get R-3-alkyl-4-heptenoic acid (C ₁-C ₃) alkyl ester (V 1a).This reaction can be carried out in reaction-inert solvent, carries out but be preferably in to have only in the excessive ortho-acetate, generally reacts at high temperature (for example at 120-160 ℃).When triethly orthoacetate (142 ℃ of boiling points) when being reagent, is used reflux temperature usually.This is reflected under the acid catalyst existence and carries out.Sour as propionic acid, PIVALIC ACID CRUDE (25), 2,2, 4-dinitrophenol, dry HCl and AlCl ₃Be effectively, preferred catalyzer is a Lewis acid, preferably AlCl ₃

In another route, the ester (formula V 1a) that obtains is used the ordinary method hydrolysis, obtain corresponding unsaturated acid (formula V 1b).

The route that another can select usefulness of the present invention is also realized easily.At first, change the S-geraniol of buying (XI) into shown in the formula (XII) optically active starting raw material with ordinary method, this points out in front and will be described in detail in the preparation in the back.Then, compound (XII) changes S-3-methyl-6-heptenoic acid (X d) or S-3-methyl enanthic acid (II) respectively into, and substep is specified in down.

In the first step, the aldehyde that hydroxyl is not protected or protected (XII a or XII b) carries out witig reaction with triphenyl phosphine methylene.Generally, triphenyl phosphine methylene is freshly prepd, is formed in reactionlessness aprotonic solvent (as the mixed solvent of tetrahydrofuran (THF) and hexane) with n-Butyl Lithium by halogenation methyl triphen (what conveniently use is bromide) usually.

For example

Though temperature is not crucial in the formation of phosphine, temperature is preferably in ± 10 ℃ in ± 25 ℃ of scopes preferably.(XII a) or the aldehyde of hydroxyl protection (XII b) and phosphine form the alkene (X b) of the not protected or protection of hydroxyl in same solvent and same temperature range respectively or (X a) for the unprotected aldehyde of hydroxyl then.

In second step, slough protecting group with hydrolysis method, this is needs when having hydroxyl protecting group to exist only, the aqueous acids condition of using in the most handy the 3rd step, the 3rd step was with the Jones oxidation method primary alconol (X b) to be oxidized to acid (X d).The so-called Jones reagent of Jones oxidation method is promptly by CrO ₃HCrO with strong acid formation ₄The aqueous solution.Typical Jones reagent is by excessive dense H ₂SO ₄, about 1: 1(is by weight) CrO ₃With the water preparation, be diluted with water to desired concn then, for example, about 3M.The alcohol of alcohol (X b), protection (X a) generally be dissolved in water miscible reactionlessness organic solvent such as acetone in, they and the reaction of the Jones reagent of at least two molar equivalents.Under these conditions, any silyl ether group is hydrolyzed to alcohol (X b) very soon, is oxidized to acid (X d) then.Temperature is not crucial, and for example, 0-50 ℃ normally satisfactory, optimum room temperature, for example 17-27 ℃.

Jones oxidation begins to be oxidized to aldehyde (X c) by alcohol earlier, does not separate out usually.Isolate intermediate aldehydes if desired, need the higher oxygenant of selectivity, come oxidation alcohol as pyridinium chlorochromate.The pure intermediate aldehydes (X c) that obtains is reoxidized (with Jones reagent or other suitable reagent) and become acid (X d).

If desired; unsaturated acid (V 1b) or (X d) are with activated form (for example; chloride of acid (V 1c) or (X e); mixed acid anhydride commonly used; or with dehydration coupling agent commonly used such as dicyclohexyl carbonization two industry amine activation), become required diester compound (formula III b) according to a conventional method with immunoregulation effect with compound coupling shown in the formula (VIII).The latter uses the ordinary method hydrolysis, obtain having immunoregulation effect the diprotic acid compound (the formula III a) or its can be used as medicine salt.

R in the formula (VIII) ⁹And R ¹⁰Be (C independently of one another ₁-C ₆) alkyl, (C ₆-C ₈) methyl cycloalkyl or benzyl.

In addition, work as R ²And/or R ³When being benzyl, formula (III b) hydrogenation of compounds can be become the immunomodulator of formula I compound.In this hydrogenation, two keys are saturated by hydrogen, and benzyl (1 or 2) is by hydrogenolysis.This hydrogenation carries out in reaction-inert solvent and hydrogenation catalyst, hydrogenation catalyst can use as, nickel or precious metal; Can be carrier (Raney nickel for example, Pd/C) or DNAcarrier free (as RhCl ₃).Solvent, temperature and pressure are not crucial.Suitable solvent is not limited to lower alcohols, can comprise ethers such as diox, tetrahydrofuran (THF) or glycol dimethyl ether; Ester class such as ethyl acetate.The most handy room temperature even some heat release of this reaction does not need cooling yet, has been avoided heating or refrigerative expense like this.Pressure is not crucial, but for fear of using expensive high-tension apparatus, pressure is preferably lower than 7 normal atmosphere.Carry out hydrogenation with Pd/C, pressure 3-6 doubly is particularly suitable for this reaction to normal atmosphere.

In addition, unsaturated acid (V 1b) or (X d) can hydrogenations under the same terms that describes in detail with earlier paragraphs, obtain S-3-alkyl enanthic acid (II).Also available opposite hydrogenation/hydrolysing step is by trans-R-3-alkyl-4-heptenoic acid (C ₁-C ₃) alkyl ester (V 1a) is by S-3-alkyl enanthic acid (C ₁-C ₃) alkyl ester obtains.The method activation of last S-3-alkyl enanthic acid to describe in detail above with the coupling of above-mentioned formula (VII) dibasic acid esters, and used aforesaid method hydrogenation, obtains the immunomodulator of above-mentioned formula I compound.

Another can be selected in the route of usefulness in the present invention, with R-trans-4-alkane alkene-3-alcohol (V) is transformed into corresponding vinyl ether (IX), this is in reaction-inert solvent (preferably excessive ethyl vinyl ether), with Hg(OAc) ₂For catalyzer and ethyl vinyl ether effect obtain, temperature range 25-40 ℃, and be that reflux temperature at ethyl vinyl ether (36 ℃ of boiling points) carries out easily.The vinyl ether that obtains (IX) is heated to 140-200 ℃, generally is in high boiling point, lipophilic reaction-inert solvent such as dimethylbenzene or naphthalane, and the rearrangement of stereotaxis ground is unsaturated aldehyde (V 1d) under pressure in case of necessity.For obtaining unsaturated acid (V 1b), can be easily with so-called Jones reagent (promptly by CrO ₃H with strong acid formation ₂CrO ₄The aqueous solution) easily make this formoxy-ization.Typical Jones reagent is by the excessive vitriol oil, about 1: 1(is by weight) CrO ₃With the water preparation, be diluted with water to desired concn then, for example, about 3M.Be to generate unsaturated acid (V 1b), usually this unsaturated aldehyde (V 1d) is dissolved in the miscible reactionlessness organic solvent (as acetone) of water in, with the Jones reagent reaction of at least 1 molar equivalent.Temperature is not crucial, and for example common 0-50 ℃ is satisfactory, the optimum room temperature, for example, 17-27 ℃.

R is methyl and R in the formula I ⁴=R ⁵The crystallized form of the compound of=hydrogen is obtained by a kind of organic solvent or mixed organic solvents recrystallization.Suitable solvent is an acetone, acetonitrile/ethanol and tetrahydrofuran (THF)/ether.Aspect the product rate of recovery, solvent is acetonitrile/ethanol preferably, but aspect product purity, then acetone is better.This novel form is more stable than previous amorphous lyophilize thing certainly, its easier operation, and crypto set more, electrostatic force is much less also, and can be prepared into more accurate dosage form.

Can be used as medicine formula I or (III is the single base and the double alkali yl salt of compound a), the common solution of usable acid, be preferably the aqueous solution and alkali (as NaOH, KOH, Na ₂CO ₃Or amine) react with suitable stoichiometrical ratio and make, salt can come out with evaporation or precipitate and separate.

Formula I of the present invention and (III) compound are used as treatment Mammals (comprising the people) because the medicine of the disease that various pathogenic microorganisms, particularly gram-negative bacteria cause.Because original and clinical caused immunosuppression meeting increases the danger that infects, The compounds of this invention can also be used as the immunostimulant of Mammals (comprising the people).

Examination gesture method is with normally or be subjected to CH ₃The male mouse of this multiply test chamber, river carries out in immune the checking oneself of crossing of H/HeN.Mouse conformed five days before use, and subcutaneous respectively then (SC) or oral (PO) gives the test compound or the placebo (salt-free thermal source) of different extent of dilution (100,10,1 and 0.1 milligram/kilogram), and used volume is 0.2 milliliter.Medication is relevant with the infection microorganism used therefor: pneumobacillus is attacked poison administration in preceding 24 and 0 hours at normal mouse; Intestinal bacteria or streptococcus aureus had been subjected to immune mouse to attack poison administration in preceding 3,2 and 1 days.(IM) shoots pneumobacillus at intragluteal injection, and abdominal injection (IP) is shot intestinal bacteria or streptococcus aureus.Use 0.2 ml volumes when attacking poison.Being to write down mortality ratio after seven days during with pneumobacillus, is to write down mortality ratio after three days when attacking poison with other two kinds of microorganisms.

Cultivate sample:

Pneumobacillus, intestinal bacteria, or streptococcus aureus: for purifying, culture is to rule on brain heart infusion (BHI) agar with the frozen blood of stocking.Choosing three bacterium colonies from cultivated 18 timely plate cultures is put in 9 milliliters of BHI broth cultures.Broth culture, is got 0.2 milliliter and is drawn on several brain heart infusion agars inclined-plane after 2 hours in 37 ℃ of growths on the rotation wobbler.Cultivated 18 hours in 37 ℃ subsequently, the inclined-plane washes out with brain heart infusion agar, checks the turbidity of culture and carries out suitable dilution with Spectronic20, makes to reach LD90 attack malicious level on normal mouse.

When the present invention's (I) and (III) compound being used on the human body as anti-infection agent or immunostimulant, generally be oral administration, subcutaneous injection, intramuscular injection, intravenous injection, the administration of abdominal injection is normally taken with composition forms up to specification in the pharmacy practice.For example, can take tablet, pill, pulvis or the granule that contains vehicle (as starch, lactose, some tackiness agents etc.).Can take capsule, this capsule is that same as described above or suitable vehicle mixes mutually, again mixture is placed capsule and gets.Also can use oral suspension, solution, emulsion, syrup and elixir, they can contain seasonings and tinting material.During treatment preparation oral administration administration of the present invention, tablet or capsule contain the effective constituent of having an appointment from 50～500 milligrams, and this is applicable to most of service condition.

The doctor will decide only dosage according to the reaction and the route of administration of different age of each patient, body weight, individual patient.Best oral dosage scope is from about 1.0～about 300 milligrams/kg/day, single or equal divided dose.The dosage of non-footpath enterally administering is better from about 1.0～about 100 milligrams/kg/day; Better scope is from about 1.0～about 20 milligrams/kg/day.

Below providing example and be intended to illustrate the present invention, is limitation of the present invention and it should not seen as, and within the scope of the invention and flesh and blood, following example can carry out many variations.

Example 1

R-is trans-the 4-hexene-3-ol

Induction stirring is being housed, barrier film, thermometer and logical N ₂In 500 milliliter of three neck round-bottomed flask of pipe, put into 270 milliliters of CH ₂Cl ₂With 7.2 gram 4A type molecular sieves.Add Ti (OCH(CH by syringe ₃) ₂) (10.7 milliliters, 0.036 mole), mixture is chilled to-66 ℃, add with 10 milliliters of CH with sleeve pipe ₂Cl ₂Reduced the L-(+ of viscosity after the dilution) tartrate diisopropyl ester (10.1 grams, 0.043 mole), other adds 5 milliliters of CH ₂Cl ₂Flushing.When temperature rises to-62 ℃, add racemic trans-2-hexene-4-alcohol (3.60 gram, 0.036 mole), and with 5 milliliters of CH ₂Cl ₂Flushing.After acetone/the dry ice bath stirs 8 minutes, when temperature is reduced to-68 ℃, add tert-butyl hydroperoxide (7.18 milliliters 3M toluene solution, 0.022 mole) with syringe.Reaction mixture is warmed to-35 ℃, keeps this temperature 18 hours in refrigerator.Cooling mixture is filled into 540 milliliters of acetone and the 11 milliliters of H that are chilled to-20 ℃ with groove line filter paper ₂In the stirring the mixture of O.Mixture slowly is warmed to room temperature and stirred 20 hours.Mixture is by diatomite filtration and use CH ₂Cl ₂Washing.The filtrate and the washing lotion that merge make its evaporation without heating, get oily matter (17.62 gram).Avoid evaporation undue, because of required product volatile.Oily matter is chromatography on 176 gram silica gel, and with 4: 1 hexanes: isopropyl ether was made elutriant, monitors with thin-layer chromatography.With variable derivation pressure head being housed and having the short column of glass simple helix ring filler to distill except that desolvating.It is 69.5 ℃ (after arriving this temperature, head temperature begins to descend) that solvent is boiled to head temperature, and residue is pure title product; Thin-layer chromatography Rf0.25(3: 1 hexane: ether).When repeating this test, with more volatile 4: 1 pentanes: ether is made elutriant, and solvent is removed sooner, so that product loses and reduces to minimum during evaporation.

Example 2

R-3-methyl-4-hexenoic acid ethyl ester

Precedent product (0.4 gram), triethly orthoacetate (3 milliliters) and trimethylacetic acid reflux 2.5 hours, cooling, and with complete reaction mixture twice of chromatography on silica gel, begin with 6: 1 hexanes: the ether wash-out, get 0.45 gram product, use 7: 1 hexanes then: the ether wash-out gets pure title product 0.35 gram; The IR(film) 2960,1736,1456,1367,1334,1280,1232,1172,1028,962,840 centimetre ^-1

In addition, contain pure S-trans-pentane/ether post flow point of 2-hexene-4-alcohol all directly introduces this step, during beginning pentane and ether steamed from reaction mixture.

Propionic acid replaces trimethylacetic acid, obtains practically identical result.Exsiccant HCl and 2,2, 4-dinitrophenol also can successfully be used as acid catalyst, but in next example, AlCl ₃It is best catalyzer.

Example 3

R-3-methyl-4-heptenoic acid

Method A

Get in 35 milliliters of refining bottles of device at the Soxhlet that has magnetic agitation and 4A type molecular sieve is housed in sleeve pipe, add precedent product (1.5 grams, 0.015 mole).Add triethly orthoacetate (9 milliliters, 0.049 mole), add AlCl again ₃(0.12 gram, 0.009 mole), mixture refluxed 2 hours on the Soxhlet extractor, at this moment between in the thin-layer chromatography demonstration, be transformed into intermediate R-3-methyl-4-hexenoic acid ethyl ester fully; Thin-layer chromatography Rf0.75(4: 1 hexane: ethyl acetate), Rf0.85(15: 5: 2 hexanes: ether: acetic acid).

Reaction mixture with the dilution of 15 milliliters of 2N NaoH and 12 ml methanol, at room temperature stirred 27 hours, at this moment between in, thin-layer chromatography shows that hydrolysis is complete.Remove methyl alcohol from reaction mixture, with 12 milliliters of H ₂The O dilution is with 3 * 25 milliliters of CH ₂Cl ₂Extract the CH of merging ₂Cl ₂With 1 * 25 milliliter of 2N NaoH back scrubbing, combining water layer and backwashings transfer to PH1 with dense HCl, with 3 * CH ₂Cl ₂Extract.Merge organic layer, use MgSO ₄Dry also evaporation gets oily title product 1.28 grams (60%); Thin-layer chromatography Rf0.65(15: 5: 2 hexanes: ether: acetic acid);

¹H-NMR(CDCl ₃) δ (ppm) 9.4(S, 1H ,-CO ₂H), 5.5(m, 2H ,-CH=CH-), 3.0-1.6(m, 5H), 1.3-0.8(m, 6H); The IR(film) 3400-2400,2960,2925,2860,1708,1458,1410,1380,1295,1228,1190,1152,1100,930 centimetres ^-1

Method B

Precedent ester products (0.24 gram) and 25 milliliters of CH ₃OH and 11.5 milliliters of 1N NaOH merge, and mixture was at room temperature stirred 3.5 hours, and during this period of time thin-layer chromatography shows that hydrolysis is complete.With 2 * 35 milliliters of ether extraction mixtures, transfer to PH2 with dense HCl, and with 3 * 35 milliliters of ether extraction.Merge acid extraction liquid, dry and evaporation obtains and corresponding to title product 0.20 gram of method A product.

Example 4

S-3-methyl enanthic acid ethyl ester

Example 2 products (0.20 gram) restrain the 5%Pd/C(50% water-wet with 0.20 in 40 milliliters of ethyl acetate), in the Paar hydrogenation equipment, hydrogenation is 3 hours under 4 atmospheric hydrogen.Reclaim catalyzer by diatomite filtration.Evaporated filtrate gets title product.

Example 5

The S-3-methyl enanthic acid

Method A

The product of example 3 (0.20 gram) is in 40 milliliters of ethyl acetate, and is moistening with 0.2 gram 5%Pd/C(50% water) in Pa Er (Paar) hydrogenation equipment, hydrogenation is 3 hours under 4 atmospheric hydrogen.Reclaim catalyzer by diatomite filtration, and evaporated filtrate, 0.2 gram buttery title product got.As needs, can under high vacuum, distill, get pure title product, boiling point 77-79 ℃/0.2 millimeter; ¹H-NMR) CDCl ₃) δ) ppm): 12.0(S, COOH), 1.0(d ,-CH ₃), 0.6-2.8(m, remaining 13H); The IR(film) 3400-2400,2960,2925,2860,1708,1458,1410,1380,1295,1228,1190,1152,1100,930 centimetres ^-1; (α)=-6.41 ° (c=1% is in methyl alcohol); n ^22.5 _D=1.427.

Method B

Press example 3 method B with the hydrolysis of precedent product, get title product.

Example 6

S-3-methyl oenanthyl chloro

The acid product of precedent or example 27 (8.5 grams, 0.062 mole) is dissolved in 18 milliliters of CH ₂Cl ₂In.Add oxalyl chloride (5.36 milliliters, 7.80 grams, 0.0614 mole) in this solution and mixture was placed 4 hours, during this period of time confirm that there to be gas to overflow again reaction is completely.This solution of acid chloride is directly used in example 8 method C immediately.Perhaps this acyl is every tells after boiling off solvent, is used for example 8 method A, and if desired, retortable to be further purified, boiling point is 45 °/1.5 millimeters.

Example 7

R-3-methyl-4-heptene acyl chlorides

The acid product (0.747 gram, 5 mmoles) of example 3 is transformed into the CH of title product with the precedent method ₂Cl ₂Solution, and be directly used in following example 9.Perhaps evaporation reaction mixture gets title product, if desired, it under reduced pressure can be steamed.

Example 8

N-(S-3-methyl oenanthyl)-D-γ-Gu Anxianji (α benzyl ester)-glycyl-D-alanine benzyl ester.

Method A

660 milligrams of (4.06 mmole) S-3-methyl oenanthyl chloros are added to 1.0 gram (2.03 mmole) D-gamma-glutamyls (in 50 milliliters of dichloromethane solutions of α benzyl ester-glycyl-D-alanine benzyl ester hydrochloride (preparation 4) and 616 milligrams of (6.09 mmole) triethylamines, and at room temperature to be stirred reaction mixture 80 hours.Steam methylene dichloride in a vacuum, its residue is dissolved in the ethyl acetate.Gained solution is used 2.5% hydrochloric acid successively, water, 10% solution of potassium carbonate, water and sodium chloride solution washing.Tell organic phase, by dried over mgso and concentrated in a vacuum.Residue grinds with ether and filters under nitrogen, gets title product, and this product all is directly used in example 10 method A.

Method B

With preparation 4 product (0.75 gram, 1.53 mmoles), 5 milliliters of methylene dichloride and triethylamine (0.212 milliliter, 1.53 mmoles) merge, and beg down at nitrogen and stir, and add S-3-methyl enanthic acid (example 5,0.20 grams, 1.39 mmoles) at 4 milliliters of CH ₂Cl ₂In solution, add dicyclohexyl carbodiimide (0.286 gram, 1.37 mmoles) then, reaction mixture stirred 16 hours.Filter reaction mixture, evaporated filtrate is dissolved in residue in 10 milliliters of ethyl acetate, and solution is used 5 milliliters of 2.5%HCl successively, 5 ml waters, 5 milliliters of 10%K ₂CO ₃MgSO is passed through in solution and the washing of 5 mL of saline ₄Drying gets 71 milligrams of (88%) title product.

Method C

Agitator is being housed, thermometer, dropping funnel and N ₂In 500 milliliter of four neck round-bottomed flask of inlet tube, the product of preparation 4 (32.8 grams, 0.059 mole) is dissolved in 175 milliliters of CH ₂Cl ₂In, and be chilled to 0-5 ℃.Remain in this temperature range, in 15 minutes, slowly splash into triethylamine (24.7 milliliters, 17.9 grams, 0.177 mole, 3 equivalents).Under the ice-water bath cooling, in 15 minutes, add the CH of the whole S-3-methyl oenanthyl chloros that obtain by example 6 ₂Cl ₂Solution, its temperature rise to 21 ℃.Continue to stir 45 minutes in ice-water bath, mixture during this period of time as a form of gel becomes too thick makes stirring that difficulty take place.Gelled block is smashed and and 125 milliliters of 10%HCl and 50 milliliters of CH ₂Cl ₂Mix.Tell organic layer, use 2 * 125 milliliters of HO successively, 2 * 125 milliliters of 10%K ₂CO ₃With 1 * 125 milliliter of H ₂MgSO is passed through in the O washing ₄Drying and evaporation get the moist white solid of 82.3 grams.This solid is dissolved in the ethyl acetate of 500 milliliters of heat.Slowly be chilled to room temperature, the crystallization of this subject product is slow, and mixture is again with other 40 milliliters of ethyl acetate dilution, so that keep good stirring.Filter and, obtain pure title product 31.1 grams, (90.5%) in 40 ℃ of following vacuum-dryings.

¹H-NMR（CDCl ₃）δ（ppm）：8.4-8.1（m，3H），7.15（s＜10H），5.1（s，4H），4.4-4.2（m，2H），3.7（d，2H），2.2（t，2H）2.1-1.7（m，6H），1.4-1.1（m，10H），0.92-0.8（m，6H）。

Example 9

N-(R-3-methyl-4-heptene acyl group)-D-γ-Gu Anxianji-(α benzyl ester)-glycyl-D-alanine benzyl ester

Press precedent method C, make preparation 4 product (2.77 grams, 5 mmoles) and the CH of whole acyl chlorides of obtaining by example 7 ₂Cl ₂Solution reaction.Evaporation is dissolved in ethyl acetate with residue again through washing and dried organic layer, revaporization, and repeatedly repeat this step, and obtain product.

Example 10

N-(S-3-methyl oenanthyl)-D-γ-Gu Anxianji-glycyl-D-L-Ala.

Method A

Whole products of example 8 method A are dissolved in 65 ml methanol.Add palladium hydroxide (250 milligrams) to this solution, mixture shook 3 hours in 4 normal atmosphere hydrogen.Leach catalyzer also in a vacuum except that desolvating.Water-soluble and the lyophilize of residue gets the purpose product.

NMR(DMSO-d ₆) show, at 8.27-8.03(m, 3H), 4.32-4.1(m, 2H) 3.72(d, J=6Hz, 2H), and 2.22(t, J=8Hz, 2H), and 2.27-1.68(m, 6H), 1.42-1.0(m 10H) and 0.94-0.8(m, 6H) locates to absorb.

When using example 8 products (0.50 gram), 90 milligrams of 20%Pd/(OH) ₂When/C(31% water-wet) carrying out in 25 ml methanol, this method gets 0.24 gram same soft have electrostatic title product, Ir(Nujol mull) 3300,2940,1740,1650,1540,1468 and 1380 centimetres ^-1; Except that latter two peak, all peaks are wide and distinguishing difference.

Method B

The product of example 8 method C (30.8 gram) and dehydrated alcohol place 2 liters of autoclaves to stir pulp for 300 milliliters, add the 5%Pd/C(1.54 gram, 50% water-wet).Mixture hydrogenation 1 hour under 4 normal atmosphere, during this period of time absorption of hydrogen is complete.Catalyzer can pass through filter paper earlier, then by 0.45 micron aperture nylon filtered and recycled, with 100-150 milliliter washing with alcohol.Merging filtrate and washing lotion, evaporation gets moist white solid, it is dissolved in the mixture of 1: 10 dehydrated alcohol of 150 milliliters of heat and acetonitrile, and the heat filter makes its clarification, evaporation, make its volume reduce to 35 milliliters, slowly be chilled to room temperature, pulverize and filter, title product that must not have electrostatic compact crystal shape weighs 20.1 grams (94%), uses the IR(Nujol mull) identify, the spike that its main distinguishing is good has 3340,3300, and 2900,2836,1725,1650,1628,1580,1532,1455,1410,1370,1280,1240,1216 and 1175 centimetres ^-1

Method C

Directly will place 1000 milliliters of acetone by the crystallized product (9.4 gram) of preceding method B preparation, reflux made its dissolving in 1 hour.Solution is chilled to room temperature, brings out crystallization with micro-method B products therefrom as crystal seed.Stir after 6 hours, the title product after being further purified is collected after filtration, with small amount of acetone washing, 35 ℃ of dryings in a vacuum, 7.25 grams, it has consistent infrared peak value with method B with the product of acetonitrile/alcohol crystal.

Method D

Precedent product (0.50 gram) and 0.026 restrains the 5%Pd/C(50% water-wet) and 125 milliliters of dehydrated alcohols place Pa Er (Paar) hydrogenation bottle.Under 4 atmospheric zone hydrogen with mixture hydrogenation 2.5 hours.Filtering recovering catalyst, the filtrate evaporation, the solid title product that must glue is directly carried out crystallization by preceding method.

Example 11

N-(R-3-methyl-4-heptene acyl group)-D-γ glutamyl-glycyl-D-L-Ala

Example 9 products (1 gram) are dissolved in 5 milliliters of CH ₃Among the OH.Add 1N NaOH(2.50 milliliter), mixture at room temperature stirred 3 hours.Boil off methyl alcohol, moist residue is diluted with 7.5 ml waters, with 2 * 7.5 milliliters of ethyl acetate extraction, is acidified to pH3.0 with 1N HCl.Extract continuously with the ethyl acetate of new preparation through acidified water, vaporize draw liquid gets title product, presses example 10 method A, this title product can be made cryodesiccated product.

Example 12

R-is trans-4-teracrylic acid-alcohol

With the method for example 1, just adding reagent elementary reaction temperature at all is-20 ℃, and the title compound of preparation 5 (10 grams, 0.088 mole) can change this title product into.After adding whole reagent, mixture stirred 1.5 hours at-20 ℃, was warmed to room temperature then, dripped 25 milliliters of HO successively and 6 milliliters of 30%NaOH(are saturated with NaCl) mixture is cooled off suddenly.The refrigerative mixture stirred 20 minutes, with 90 milliliters of CH ₂Cl ₂Dilution divides water-yielding stratum, with the CH of 2 * 50 milliliters of new preparations ₂Cl ₂Washing.Merge organic layer, through MgSO ₄Dry and to be evaporated to residual volume be 50 milliliters, chromatography on silica gel, earlier with behind the pentane with 4: 1 pentanes: ether is made elutriant.Discard stream part of the beginning that contains small amount of polar impurity.Merging Rf is 0.3(4: 1 pentane: ether) pure products part and evaporation get title product 5.6 grams, the clarifying colourless liquid of this product.

Example 13

R-3-ethyl-4-heptenoic acid

With example 3 method A, just when separating, use substituted ether CH ₂Cl ₂, precedent product (5.7 grams, 0.05 mole) and ethyl orthoacetate condensation are reset and saponification, get the buttery title product, thin-layer chromatography Rf0.4(2: 1 hexane: ether).With the skim system, the Rf of intermediate ethyl ester is 0.25.

Example 14

S-3-ethyl enanthic acid

Precedent product (1.4 gram) is at 20 milliliters of CH ₃Among the OH with 0.1 gram 5%Pd(OH)/C hydrogenation 1 hour under 4 normal atmosphere, filtering recovering catalyst.From filtrate, boil off solvent and, get title product 1.0 grams the residue distillation; Boiling point 76-77 ℃/0.4 torr; (α) ²⁵ _D-1.6 ° (C=2% is in methyl alcohol).

Example 15

S-3-ethyl-oenanthyl chloro

At dry N ₂Precedent product (1.0 grams, 0.00633 mole) is dissolved in 10 milliliters of CH down, ₂Cl ₂In, adding oxalyl chloride (0.547 milliliter, 0.00627 mole), mixture stirred 1 hour, during this period of time stopped evolving gas.At N ₂Flow down the solution evaporation to 7 of gained title product milliliter.Just can be directly used in next step.

Use same quadrat method, can be with the solution of example 13 undersaturated acid changing into R-3-ethyl-4-heptene acyl chlorides.

Example 16

N-(S-3-ethyl oenanthyl)-D-γ-Gu Anxianji (2-benzyl ester)-glycyl-D-alanine benzyl ester

The title product (1.1 grams, 0.00222 mole) of preparation 4 is dissolved in 30 milliliters of CH ₂Cl ₂And in the triethylamine (0.935 milliliter, 0.00666 mole).CH with the S-3-ethyl oenanthyl chloro (2.3 milliliters, 0.00211 mole) of precedent ₂Cl ₂Solution adds, and mixture is at N ₂Under stirred 0.75 hour, use 20 milliliters of 10%HCl successively, H ₂O, 10%K ₂CO ₃With the saturated brine washing, through MgSO ₄Drying, evaporation gets solid residue, and grinding also with ether, filtered and recycled gets 0.8 gram.It is dissolved in the ethyl acetate, once more with top method washing, evaporation once more, solid 0.7 restrains again, chromatography on silica gel, with 97: 3CHCl: CHOH makes elutriant, gets 467 milligrams of pure title product.

Use same quadrat method, the title product coupling with 3-ethyl-4-heptene acyl chlorides and preparation 4 gets N-(R-3-ethyl-4-heptene acyl group)-D-γ-Gu Anxianji (α benzyl ester)-glycyl-D-alanine benzyl ester.

Example 17

N-(S-3-ethyl oenanthyl)-D-γ-Gu Anxianji-glycyl-D-L-Ala

Precedent title product (467 milligrams) is pressed example 14 method hydrogenations.After reclaiming catalyzer, the filtrate evaporation gets foam, makes its soluble in water and filtration, and lyophilize gets 238 milligrams of title product.

¹H-NMR（DHSO-d ₆）δ（ppm）：8.20-8.00（m，3H），4.24-4.16（m，2H），3.74-3.60（m，2H），2.18（t，J＝7，2H），2.02（d，J＝7，2H），2.02-1.60（m，3H），1.26（d，J＝6，3H），1.26-1.08（m，8H），0.92-0.74（m，6H）。

Use the same method, the unsaturated product of precedent can be transformed into same product.

Example 18

N-(R-3-ethyl-4-heptene acyl)-D-γ-Gu Anxianji-glycyl-D-L-Ala

Can be with the unsaturated product example 11 method hydrolysis cost title product of example 16.

Example 19

N-(S-3-ethyl oenanthyl)-D-γ-Gu Anxianji (α benzyl ester)-glycyl-D-L-Ala butyl ester

Method with example 17; S-3-ethyl oenanthyl chloro and D-γ-Gu Anxianji (α benzyl ester)-glycyl-D-L-Ala butyl ester (1.0 grams, 0.00222 mole) coupling with example 16 obtain this title product; separate and purifying with identical method, get 0.7 gram.

¹H-NMR（DMSO-d ₆）δ（ppm）：8.22（d，J＝7，1H），8.12-8.00（m，2H），4.40-4.16（m，2H），4.08-3.95（m，2H），3.75-3.62（m，2H），2.18（t，J＝6，2H），2.02（d，J＝6，2H），2.04-1.62（m，3H），1.60-1.46（m，2H），1.38-1.10（m，15H）0.90-0.75（m，9H）。

Use the same method, R-3-ethyl-4-heptene acyl chlorides can be transformed into N-(R-3-ethyl-4-pentenoyl)-D-γ-Gu Anxianji (α benzyl ester)-glycyl-D-L-Ala butyl ester.

Example 20

N-(S-3-ethyl oenanthyl)-D-γ-Gu Anxianji-glycyl-D-L-Ala butyl ester

With the method for example 14,, get this title product with the hydrogenation of precedent title product.After reclaiming catalyzer, evaporated filtrate gets solid, grinds and filters with ether, gets 256 milligrams, fusing point 130-131 ℃.

Use the same method, can change the unsaturated product of precedent into same product.

Example 21

R-3-ethyl-4-heptenal

With example 12 R-trans-300 milliliters of dry ethyl vinyl ether vlil of 4-teracrylic acid-alcohol (3.0 gram), gradation adds five parts of Hg(OAc in this process) ₂(every part 2 gram, in the time of 2 hours at interval in) after refluxing 18 hours in addition, is cooled to room temperature with clear liquor, with 0.5 milliliter of Glacial acetic acid processing and stirred 3 hours.With ether dilution gained solution, pour in 200 milliliters of 5%KOH solution, use ether extraction three times.Use K after merging ether extracted liquid ₂CO ₃Drying, evaporation, allyl vinyl ether, R-is trans-0-vinyl-4-teracrylic acid-alcohol.

The solution of above-mentioned allyl vinyl ether (2.3 gram) in 20 milliliters of naphthalanes refluxed 10.5 hours.Then gained solution is cooled to room temperature; Directly place chromatography on the silicagel column.After removing naphthalane with hexane flushing, with ether wash-out post and evaporation, title product.

Example 22

S-3-ethyl-4-heptenoic acid

Precedent product (100 milligrams) is dissolved in 4 milliliters of acetone and is chilled to 0-5 ℃.In another flask, CrO ₃(72.1 grams, 0.72 mole) and 50 milliliters of H ₂O mixes, and 0-5 ℃ of stirring, slowly adds 62.1 milliliters of dense H ₂SO ₄, mixture H ₂O is diluted to 250 milliliters, gets 2.88 moles of H ₂CrO ₄Solution (Jones reagent).In the acetone soln above gradation is added in 1 hour with a solution (1 milliliter).Add fashionable temperature at reagent and rise, and make temperature remain on 17-25 ℃.Mixture is chilled to 6 ℃ again, (temperature allows to rise to 20 ℃ during this period) adds 70 milliliters of 2-propyl alcohol in 10 minutes, concentrates under vacuum then, gets oily matter, under agitation in 50 minutes, in oily matter, add 8 milliliters of 2.5N NaoH, keep 22 ± 5 ℃ of temperature.Mixture heating up is also passed through diatomite filtration, with the NaoH washing of 2.5N heat.Merging filtrate extracts with 3 * 300 milliliters of isopropyl ethers.Merge organic layer, with 200 milliliters of anti-extractions of 2N NaoH.Combining water layer slowly adds 0.5 milliliter of dense HCl water layer is acidified to pH1, and product is extracted in the isopropyl ether of 3 * 300 milliliters of new preparations.Organic layer merges and evaporation, gets title product, is oily matter.

Example 23

S-7-(tertiary butyl dimethyl methyl silica alkoxyl group)-5-methyl isophthalic acid-heptene

Agitator is being housed, and thermometer advances N ₂In 500 milliliter of four neck round-bottomed flask of pipe and feed hopper, first base three phenyl phosphonium bromides (25.7 grams, 0.072 mole, 1.25 equivalents) are with 77 milliliters of THF(tetrahydrofuran (THF)s) stir pulp, and in acetone/wet ice bath, cool off.N-Butyl Lithium (43.2 milliliters of 1.6N hexane solutions, 0.069 mole, 1.20 equivalents) is put into feed hopper, butyllithium is added initial temperature in 1 hour in-8 ℃ the soup compound, and temperature rises and maintain ± and 1 ℃.Mixture stirred 0.5 hour at 0-2 ℃ again, was completed in the thin suspensoid of LiBr to guarantee intermediate methylene tri Phenylphosphine.Added the solution of aldehyde product (14.1 grams, 0.0576 mole) in 14 milliliters of THF of preparation 2 in 40 minutes, controlled temperature is at 3-7 ℃ in batches.Behind the restir 15 minutes, use the thin-layer chromatography inspection, with 3: 1 hexanes: ether was a developping agent, proved (the initial aldehyde Rf0.6 that do not have initial aldehyde; Product Rf0.95).Reaction mixture is warmed to room temperature also with 150 milliliters of ethyl acetate and 90 milliliters of H ₂The O dilution.Organic layer is told and with 2 * 100 milliliters of H ₂The O washing.With 40 milliliters of counter three parts of water layers that extract merging of ethyl acetate.Merge organic layer, through MgSO ₄Drying, evaporation gets 25 gram oily matter, grinds with 10 milliliters of hexanes, filters on sintered glass plate funnel, with 4 * 10 milliliters of hexane thorough washing solids, merges hexane filtrate and washing lotion, evaporation; Getting title product 13.5 grams (96.6%) is oily matter.

¹H-NMR(CDCl ₃) δ (ppm): 5.4-6.2(m ,=CH), 4.8-5.3(m ,=CH ₂), 3.7(t, J=6.5H ,-OCH ₂O-), 0.08(s, C(CH ₃) ₃) and 0.0(s, Si(CH ₃) ₂), be mixed with 8% mole of (C ₆H ₅) ₃PO(7.6, s, 1.25H).

Example 24

S-3-methyl-heptan-6-alkene-1-alcohol

Method A

Use the precedent method, but, prepare 6 aldehyde product (26.3 grams, 0.20 mole with first base three phenyl phosphonium bromides and each 2.2 equivalent of n-Butyl Lithium; Purity is qualified) react with the methylene tri Phenylphosphine.Though formed gluing solid when adding aldehyde solution, as long as reactant is warmed to room temperature, reactant just becomes rare soup compound.With 500 milliliters of H ₂O and 300 milliliters of ethyl acetate diluted reaction mixtures.Tell each layer, and with 3 * 250 milliliters of H ₂The O washing.The water layer that merges is with 2 * 300 milliliters of ethyl acetate backwashes.Three parts of organic layers merge, through MgSO ₄Drying and evaporation get 65.7 gram oily matter, wherein contain 25.6 gram (100%) title product and about 40 and restrain triphenyl phosphine oxides, can further handle by the method for following example 26.

Method B

By with the hydrolysis according to a conventional method of example 23 products, for example, can more easily obtain pure title product by the method for back example 26 described hydrolysis in dilute sulphuric acid.This title product is extracted in the ethyl ester ethyl ester, through MgSO ₄Drying and evaporation can be separated title product.

Example 25

S-3-methyl-6-heptenal

The title product (1.14 grams, 0.01 mole) of previous examples is dissolved in 20 milliliters of CH ₂Cl ₂In, be cooled to 0 ℃.Pyridinium chlorochromate (400 grams, 0.02 mole) is added in batches, and simultaneous temperature remains on 0-5 ℃.Mixture is warmed to room temperature, stirs 2 hours, filters by a silicagel pad, and evaporated filtrate gets the oily title product, further purifies as the available distillation method of needs.

Example 26

S-3-methyl-6-heptenoic acid

Method A

Stirring is being housed, and temperature is taken into account in 2000 milliliters of three-necked bottles of addition funnel, the title product of example 23 (81 grams, purity is qualified, 0.33 mole) is dissolved in 400 milliliters of acetone, and is cooled to 0-5 ℃.In another flask, with CrO ₃(72.1 grams, 0.72 mole) mixes with 50 ml waters, and under 0-5 ℃ of stirring, 62.1 milliliters of vitriol oils is slowly splashed into, and with 250 ml water diluted mixture things, promptly obtains 2.88 moles of H ₂CrO ₄Solution (Jones reagent).In 1.2 hours, back solution (240 milliliters, 0.67 mole) is added in the top acetone soln in batches.Temperature rises to 17 ℃ very soon, remain on 17-25 ℃ when reagent adds fashionable temperature, add to the end, also do not cause temperature rise, mixture is chilled to 6 ℃ once more, in 10 minutes, add 70 milliliters of 2-propyl alcohol (this moment, temperature rose to 20 ℃), be condensed into oil then in a vacuum, under keeping 22 ± 5 ℃ of temperature and stirring, in 50 minutes, 400 milliliters of 5N NaoH are added, with 400 milliliters of H ₂O dilution heavy-gravity reaction mixture filters on diatomite, will wet cake again with 400 milliliters of H ₂O and 50 milliliters of 5N NaoH furnishing pulpous states heat in vapor bath, and heavy filtration is with the filtrate of 3 * 300 milliliters of isopropyl ether washing merging.With 200 milliliters of anti-organic layer solution that extract merging of 2N NaoH, slowly add 50 milliliters of concentrated hydrochloric acids so that the water layer solution that has merged is acidified to pH1.0, product is extracted in 3 * 300 milliliters of new preparation isopropyl ethers.Merge organic extracting solution, evaporation gets title product 29.1 grams (61%), is oily matter.Diatomite filter cake extracting solution is alkalized once more,, obtain product 7.7g(16% in addition) through separating with quadrat method.The merging product ¹H-NMR(CDCl ₃), δ (ppm): 11.9(S ,-COOH), 5.8(m ,=CH), 5.0(m ,=CH ₂), 1.0(d ,-CH ₃), the isopropyl ether peak is 3.7 and 1.1, indication is mixed with the 8.6%(mole) (6.3%(weight)) isopropyl ether, but do not influence this product in further operational application.

Method B

According to the product of this case method A oxidation example 24 method A (65.7 grams contain 26.3 grams, 0.20 mole of S-3-methyl heptan-6-alkene-1-alcohol).At the adding Virahol and after stirring 1 hour under 0-5 ℃, reaction mixture is complete green, boil off organic solvent, dilute aqueous residue with 250 ml waters, extract with 3 * 250 milliliters of isopropyl ethers, merge organic extracting solution, handle, generate a large amount of throw out (C with 160 milliliters of 2N NaoH ₆H ₅) ₃PO(mixes in raw material), reclaim after filtration, with 1N NaoH washing, merging filtrate and washing lotion, solution layering, organic layer is more in addition with 80 milliliters of 1N NaoH washings, merge all water layers,, be acidified to pH1.0 with 50 milliliters of dense HCl with 3 * 250 milliliters of isopropyl ether washings, required product is extracted in the isopropyl ether of 3 * 250 milliliters of new preparations, and acid organic extracting solution of merging is with MgSO ₄Drying, evaporation gets title product 14.0 grams, and as needs, available distillation method is further purified.

Example 27

The S-3-methyl enanthic acid

10%Pd/C(1.64 is restrained, contain 50% water), (3.28 grams of unsaturated acid in 150 milliliters of ethyl acetate and the precedent, purity is qualified) in the Parr hydrogenation bottle of packing into, pulpous state liquid carries out hydrogenation under 4 normal atmosphere, absorption of hydrogen needs 1.5 hours fully approximately, reclaims catalyzer through diatomite filtration, filtrate gets title product 3.20 grams (96%) after evaporation, be oily matter.

In addition, 5%Pd/C(2 gram contains 50% water), the title product (33.3 grams, purity is qualified) of precedent and 150 milliliters of ethyl acetate pack in 1 liter of autoclave together, at 30-31 ℃, hydrogenation is 2 hours under 4 barometric points, this moment H ₂Be absorbed fully.Carry out filtering recovering catalyst on diatomite, evaporated filtrate gets 35.4 gram oily matter, distills under high vacuum, gets pure title product 29.6 grams (87.6%); Boiling point 77-79 ℃/0.2 millimeter; ¹H-NMR(CDCl ₃) δ (ppm): 12.0(s ,-COOH), 1.0(d ,-CH ₃), 0.6-2.8(m, remaining 13H); The IR(film) 3400-2400,2960,2925,2860,1708,1458,1410,1380,1295,1228,1190,1152,1100,930 centimetres ^-1, (α) ²⁵ _D=-6.41 ° (C=1% is in methyl alcohol), n ^22.5 _D=1.427.

Example 28

S-3-methyl-6-heptene acyl chlorides

The product acid (0.747 gram, 5 mmoles) of example 4 is changed the CH of cost title product with the method for example 6 ₂Cl ₂Solution can directly apply in the following example 9.In addition,, can obtain title product, if desired, title product under reduced pressure can be distilled the reaction mixture evaporation.

Example 29

N-(S-3-methyl-6-heptene acyl group)-D-γ-Gu Anxianji-(α benzyl ester)-glycyl)-the D-alanine benzyl ester

With example 8 method C, with preparation 4 product (2.77 grams, 5 mmoles) and the whole acyl chlorides that obtain from example 28 at CH ₂Cl ₂Middle coupling.Initial product 2.77 grams are obtained by evaporation washing lotion and the dry organic layer of crossing, and this product is dissolved in 20 milliliters of ethyl acetate, with 20 milliliters of hexane dilutions, cooling, obtain pure product 2.24 grams (77%) after filtration, fusing point 137.5-139.5 ℃.

Example 30

N-(S-3-methyl-6-heptene acyl group)-D-γ-Gu Anxianji-glycyl-D-L-Ala

The product (1 gram) of example 29 is dissolved in 5 ml methanol, adds 1N NaoH(2.50 milliliter), at room temperature stirred the mixture 3 hours, evaporation methyl alcohol, with 7.5 milliliters of HO dilution water residues,, be acidified to pH3.0 with 1N HCl with 2 * 7.5 milliliters of ethyl acetate extraction.Ethyl acetate with new preparation is extracted acidic aqueous solution continuously, and vaporize draw liquid gets title product, according to example 10 method A, title product can be transformed into the lyophilize thing.

Example 31

N-(S-3-methyl-6-heptene acyl group)-D-gamma-glutamyl (α-benzyl ester)-glycyl-D-L-Ala butyl ester

With example 8 method A, make D-gamma-glutamyl (α-benzyl ester)-glycyl-D-L-Ala butyl ester (preparation 4) and S-3-methyl-6-heptene acyl chlorides coupling, get this title product.

Example 32

N-(S-methyl oenanthyl)-D-gamma-glutamyl-glycyl-D-L-Ala butyl ester

With the method hydrogenation of example 10, the precedent product can be changed the cost title product.

Preparation 1

Racemization is trans-the 4-hexene-3-ol

400 milliliters of THF are packed into be furnished with stirring, addition funnel, temperature is taken into account in 3 liter of four neck flask of nitrogen inlet pipe, and THF is cooled to-70 ℃, adds ethyl-magnesium-bromide (the THF solution of 600 milliliters of 2M, 1.2 moles).Maintain the temperature at-70 ℃ to-60 ℃, in 7 minutes, crotonaldehyde (78 milliliters, 0.94 mole) is added in-70 ℃ and stirs after 20 minutes, make mixture slowly be warming up to-20 ℃, stirred 1 hour, mixture is cooled to-70 ℃ once more, adds 200 milliliters of saturated NH carefully ₄Cl makes quenching (beginning to bubble), is warming up to room temperature, and with 200 milliliters of acetate and the dilution of 100 ml waters, two-layer liquid is saturated with NaCl.Divide water-yielding stratum, with 2 * 500 milliliters of ether extraction.Merge initial organic layer and ether extracted liquid,, use MgCO with 4 * 250 milliliters of saturated NaHCO washings ₄Drying, evaporation, under atmospheric pressure distillation leftover gets 69 grams; Boiling point 133-137 ℃., ¹H-NMR(CDCl ₃) δ (ppm): the m of 5.53(complexity, 2H), 3.39(m, 1H), 2.26(s, 1H), 1.71(d, 3H), 1.43(m, 2H), 0.84(t, 3H).

Preparation 2

Glycyl-D-alanine benzyl ester hydrochloride

12.3 gram (60 mmole) dicyclohexyl carbodiimides addings are contained 10 gram (57 mmole) N-tert.-butoxy-carbonyl glycines, in 100 milliliters of cold (0 ℃) methylene dichloride of 20 gram (57 mmole) D-alanine benzyl ester tosilate and 5.77 gram (57 mmole) triethylamines, make reaction mixture be warming up to room temperature, after 18 hours mixture is filtered, in a vacuum concentrated filtrate.Residue is dissolved in 200 milliliters of ethyl acetate, uses 2.5% hydrochloric acid, water, and saturated sodium bicarbonate and salt brine solution washing organic layer are told organic layer, use dried over mgso, vapourisation under reduced pressure.To add in the resulting oil with 200 milliliters of saturated dioxs of hydrogenchloride.Add 400 milliliters of ether after 30 minutes, filtration product under nitrogen gas stream gets 10.9 grams (70% productive rate).

Preparation 3

N-tert-butoxycarbonyl-D-γ-Gu Anxianji (α-benzyl ester) hydroxyl succinic diamide ester

30.9 gram (15 mmole) dicyclohexyl carbodiimides are added in 1500 milliliters of methylene dichloride that contain 50 gram (143 mmole) N-tert-butoxycarbonyl-D-gamma-glutamic acid α-benzyl esters and 17.3 gram (150 mmole) N-hydroxyl succinic diamides, and at room temperature the reaction mixture with gained stirred 18 hours.Cross filter solid, in a vacuum concentrated filtrate.Grind residue with ether, filter collection solid gets 43.7 grams (68% productive rate) under nitrogen gas stream.

Preparation 4

D-γ-Gu Anxianji (α-benzyl ester)-glycyl-D-alanine benzyl ester hydrochloride

At room temperature; to contain 4.3 gram (9.45 mmole) N-tert-butoxycarbonyl-D-γ-Gu Anxianji (α-benzyl ester) hydroxyl succinic diamide esters; 2.71 100 milliliters of dichloromethane solutions of gram (9.92 mmole) glycyl-D-alanine benzyl ester hydrochloride and 1.0 gram (9.92 mmole) triethylamines stirred 18 hours; concentrate in a vacuum then; residue is dissolved in 200 milliliters of ethyl acetate; solution 2.5% hydrochloric acid; water; 10% salt of wormwood and salt water washing; tell organic layer, use dried over mgso, vapourisation under reduced pressure; resistates is handled with the saturated dioxan of hydrogenchloride; stirred 2 hours, concentrated solution grinds residue to doing with ether in a vacuum; under nitrogen gas stream, filter, get solid 3.41 grams (73% productive rate).

Use the same method, product and glycyl-D-L-Ala butyl ester in the preparation of front are transformed into D-γ-Gu Anxianji (α benzyl ester)-glycyl-D-L-Ala butyl ester.

Preparation 5

Racemization is trans-4-teracrylic acid-alcohol

With preparation 1 method, but with common ether as solvent, 2-amylene (25 grams, 0.30 mole) is transformed into title product, 25.6 restrain; Boiling point 145-150 ℃.

Preparation 6

0-(tertiary butyl dimethyl methyl silicon)-the S-geraniol

At room temperature (21 ℃), with 5-geraniol (547 grams, 3.5 mole) be dissolved in 547 milliliters of DMF(dimethyl formamides) in, this solution placed agitator is housed, thermometer advances in 500 milliliter of four neck round-bottomed bottle of N pipe and addition funnel, adds imidazoles (262.1 grams, 3.85 mole, 1.1 equivalents).Temperature is reduced to 13 ℃, further reduces to-6.5 ℃ with ice/acetone bath.In 1.25 hours, will under high degree of agitation, be dissolved in 1160 milliliters of TERT-BUTYL DIMETHYL CHLORO SILANE (580.3 grams, 3.85 moles, 1.1 equivalents) among the DMF in advance and add, make temperature slowly rise to 11 ℃ simultaneously.Behind other 0.5 hour, (3: 1 hexanes: ether) indication is transformed into needed product (raw material Rf is 0.2 thin-layer chromatography fully; Product Rf is 0.9).

Mixture added in 500 milliliters of hexanes and 1000 milliliters of ice and the water be divided into two layers, with 2000 milliliters of ice-cold 0.25N salt acid elution organic layers, merge two parts of water layers, with 500 milliliters of hexane wash, this washing lotion and initial organic layer merging are with 500 milliliters of saturated NaHCO ₃MgSO is used in washing ₄Drying, evaporation, title product 986.7 gram because there is a small amount of solvent to exist, think 104% of theoretical amount, ¹H-NMR(CDCl ₃) δ (ppm), 5.2(t, J=7Hz ,=CH), 3.65(t, J=6.5Hz ,-O-CH ₂-), 1.7 and 1.65(2S, 2 * C(CH ₃)), 0.8(S ,-SiC(CH ₃) ₃), and 0.0(S ,-SiC(CH ₃) ₂),

Preparation 7

The S-6-tertiary butyl-dimethylsilane oxygen)-4-methyl-hexanal

The product (81.2 grams, solvent is proofreaied and correct, 0.3 mole) for preparing previously is dissolved in 120 milliliters of CH ₂Cl ₂And 81 milliliters of CH ₃Among the OH, this solution places mechanical stirrer is housed, logical O ₃/ O ₂The straight glass tube of stream, temperature are taken into account in outlet pipe and the 500 milliliter of four neck round-bottomed flask that saturated KI trap links to each other.Add NaHCO ₃(6.3 grams, 0.075 mole, 0.25 equivalent), mixture is cooled to-10 ℃ with acetone/the dry ice bath.Work as O ₃/ O ₂Degree of cooling again when being blown into reaction solution, maintain-72 ℃ to-75 ℃ totally 6 hours.After less than 1 hour, reaction mixture no longer absorbs O ₃, this can be proved by the yellow that forms in the KI trap.Make the complete situation of the thin-layer chromatography Indicator Reaction of developping agent with hexane, (raw material Rf is 0.3; Intermediate is Rf0.0).

Contain excessive O ₃Reaction mixture clean with N, add methyl-sulfide (26.4 milliliters, 22.4 grams, 0.36 mole, 1.2 equivalents), remove cooling bath, mixture is heated to room temperature, at N ₂Flow down and stirred 16 hours, this moment thin-layer chromatography (6: 1 hexanes: ether) indication does not have intermediate raw material to have that (intermediate raw material Rf is 0.8; Product Rf is 0.05).Evaporating mixture, residue is at 150 milliliters of ethyl acetate and 300 milliliters of H ₂Distribute among the O, with 300 ml waters washings organic layer, the water layer that has merged with 150 milliliters of ethyl acetate back scrubbing of newly preparing.Merge organic layer, use MgSO ₄Drying is evaporated, and gets oily matter, evaporates 16 hours under high vacuum at last, obtains quantitative title product 74.7 grams, and the 101.9%(of theoretical amount is owing to there is the event of a small amount of solvent impurity) ¹H-NMR(CDCl ₃) δ (ppm); 9.75(t ,-CHO), 3.6(t, J=6Hz ,-O-CH ₂-), 0.9(s ,-SiC(CH ₃) ₃), 0.0(s ,-Si(CH ₃) ₂).

Preparation 8

S-6-hydroxy-4-methyl hexanal

Method A

To magnetic stirrer is housed, in 250 milliliter of three neck round-bottomed flask that thermometer, gas inlet pipe and outlet pipe link to each other with the scrubbing bottle that contains saturated KI, add 81 milliliters of CH of S-geraniol (31.25 grams, 0.20 mole) ₂Cl ₂And 54 milliliters of CH ₃OH solution, and be cooled to-8 ℃.Maintain the temperature between-2 ℃ and-10 ℃, in 4.5 hours, be blown into O ₂/ O ₃Bubble to reactant, this moment excessive O ₃Captured by KI solution, by positive starch/complete situation of KI paper test Indicator Reaction, mixture remains on-5 ℃, uses N ₂Gas cleans, and adds methyl-sulfide (17.7 milliliters, 15.0 grams, 0.24 mole).Make reaction mixture be warming up to room temperature, stirred then 16 hours, (S-geraniol Rf is 0.7 to the complete situation of thin-layer chromatography (making developping agent with isopropyl ether) Indicator Reaction this moment; Product Rf is 0.4), the final evaporation solvent obtains 50.8 gram oily matter.With 30 milliliters of ethyl acetate and 25 milliliters of H ₂It is single phase that O dilutes this oil, adds 150 milliliters of ether again and obtains two-phase.Solution layer is separated, and with 2 * 25 ml waters washing organic phase, uses MgSO ₄Drying, evaporation gets colorless oil 23.9 grams.With the water layer that 2 * 50 milliliters of ethyl acetate extraction merge, united extraction liquid, drying, evaporation, get 7.8 grams in addition and be colorless oil.Merge colorless oil, revaporization, title product 28.3 grams (theoretical amount 108.7%).Thin-layer chromatography (top once mentioned) shows that except that being mixed with solvent, product is pure, as mentioned above, is applicable to next step reaction.

Method B

With general method, as used dilute sulfuric acid process in the top example 4, with the product hydrolysis of preparation 2.With extraction method title product is extracted in the ethyl acetate, evaporation is handled as top method A, and title product is isolated.

Claims (16)

1, prepare the method for R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid, the method includes the steps of:

(a) racemization trans-alkene-3-alcohol or trans-4-teracrylic acid-alcohol react with tert-butyl hydroperoxide 4-, reaction be have quantity to be enough to oxidation S-enantiomorph and keep unreacted trans-carry out by titanium tetraisopropylate and L-(+)-tartrate diisopropyl ester existence of alkene-3-alcohol or trans-R-4-teracrylic acid-alcohol for R-4-

(b) in the presence of acid, above-mentioned trans-R-4-alkene-3-alcohol or trans-R-4-teracrylic acid-alcohol and ortho-acetic acid three [(C ₁-C ₃) the alkyl ester condensation, get R-3-methyl-4-heptenoic acid (C ₁-C ₃)-alkyl ester or R-3-ethyl-4-heptenoic acid (C ₁-C ₃)-alkyl ester, and

(c) above-mentioned (C of hydrolysis ₁-C ₃) alkyl ester, form R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid.

2, prepare the method for 5-3-methyl-enanthic acid, the method includes the steps of:

(a) racemization trans-4-hexene-3-ol or trans-4-teracrylic acid-alcohol and tert-butyl hydroperoxide reaction, this reaction be have quantity will be enough to oxidation S-enantiomorph and keep unreacted trans-titanium tetraisopropylate and the L-(+ of R-4-hexene-3-ol or trans-R-4-teracrylic acid alcohol)-carry out in the presence of the tartrate diisopropyl ester

(b) in the presence of acid, make trans-R-4-hexene-3-ol or trans-R-4-teracrylic acid-alcohol and ortho-acetic acid three ((C ₁-C ₃) alkyl) ester condensation, get R-3-methyl-4-heptenoic acid (C ₁-C ₃) alkyl ester or R-3-ethyl-4-heptenoic acid (C ₁-C ₃) alkyl ester, perhaps

(c) above-mentioned (C of hydrolysis ₁-C ₃) alkyl ester, form R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid; And

(d) the above-mentioned 4-heptenoic acid of catalytic hydrogenation generates S-3-methyl enanthic acid or S-3-ethyl enanthic acid; Or

(e) above-mentioned (C of catalytic hydrogenation ₁-C ₃) alkyl ester, generate S-3-methyl enanthic acid (C ₁-C ₃) alkyl ester or S-3-ethyl enanthic acid (C ₁-C ₃) alkyl ester, and

(f) hydrolysis above-mentioned S-3-methyl enanthic acid ester or S-3-ethyl heptanoate generate S-3-methyl enanthic acid formula S-3-ethyl enanthic acid.

3, according to the method for claim 1 or 2, (C wherein ₁-C ₃) alkyl is an ethyl respectively, and acid is AlCl ₃

4, prepare the method for R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid, the method includes the steps of:

(a) racemization trans-4-hexene-3-ol or trans-4-teracrylic acid-alcohol and tert-butyl hydroperoxide reaction, reaction is to be enough to oxidation S-enantiomorph and to keep unreacted trans R-4-hexene-3-ol or the titanium tetraisopropylate and the L-(+ of trans-R-4-teracrylic acid-alcohol in quantity)-carry out in the presence of the tartrate diisopropyl ester

(b) at Hg(CH ₃CO ₂) ₂Exist down, make trans-R-4-hexene-3-ol or trans-R-4-teracrylic acid-alcohol and ethyl vinyl ether reaction, generate corresponding 0-vinyl ether,

(c) above-mentioned 0-vinyl ether adds thermal rearrangement in inert solvent, generates R-3-methyl-4-heptenal or R-3-ethyl-4-heptenal, and

(d) in diluted mineral acid, use the above-mentioned 4-heptenal of chromic trioxide oxidation, generate above-mentioned R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid.

5, according to the method for claim 4, wherein mineral acid is a sulfuric acid.

6, prepare the method for S-3-methyl-6-heptenoic acid, the method includes the steps of:

(a) in inert solvent; S-6-hydroxy-4-methyl-hexanal or S-6-(are protected by silyl) hydroxy-4-methyl hexanal and triphenyl phosphine methylene reaction; formation S-7-hydroxy-5-methyl base-1-heptene or S-7-(are protected by silyl) hydroxy-5-methyl base-1-heptene

(b) when step (a) contains the silyl blocking group, the hydrolysis in diluted mineral acid of this blocking group forms S-7-hydroxy-5-methyl base-1-heptene, and can be used as an independent step to carry out, or carry out simultaneously with next step, and

(c) make above-mentioned S-7-hydroxy-5-methyl base-1-heptene and chromic trioxide oxidation in diluted mineral acid, form S-3-methyl-6-heptenoic acid.

7, in accordance with the method for claim 6, wherein S-6-hydroxy-4-methyl hexanal is a reactant in step (a), and mineral acid is a sulfuric acid.

8, in accordance with the method for claim 6; wherein S-6-(is protected by silyl) hydroxyl-4-hexanal is reactant in step (a); step (b) reaches (c) carries out simultaneously, and the blocking group silyl is a t-butyldimethylsilyl, and mineral acid is a sulfuric acid.

9, prepare the method for S-3-methyl enanthic acid, the method includes the steps of:

(a) in inert solvent; S-6-hydroxy-4-methyl hexanal or S-6-(are protected by silyl) hydroxy-4-methyl hexanal and triphenyl phosphine methylene reaction; formation S-7-hydroxy-5-methyl base-1-heptene or S-7-(are protected by silyl) hydroxy-5-methyl base-1-heptene

(b) when the product of step (a) contains the silyl blocking group, make the hydrolysis in diluted mineral acid of this blocking group, form S-7-hydroxy-5-methyl base-1-heptene, can be used as an independent step carries out, or carries out simultaneously with next step,

(c) in diluted mineral acid, make above-mentioned S-7-hydroxy-5-methyl base-1-heptene and chromic trioxide carry out oxidizing reaction, form S-3-methyl-6-heptenoic acid, and

(d) make two keys in the above-mentioned 6-heptenoic acid through catalytic hydrogenation, form the S-3-methyl enanthic acid.

10, in accordance with the method for claim 9, wherein S-6-hydroxy-4-methyl hexanal is a reactant in step (a), and mineral acid is a sulfuric acid.

11, in accordance with the method for claim 9; wherein S-6-(is protected by silyl) hydroxyl-4-hexanal is reactant in step (a); step (b) reaches (c) carries out simultaneously, and the blocking group silyl is a t-butyldimethylsilyl, and mineral acid is a sulfuric acid.

12, preparation has the method for absolute stereo chemistry formula I compound,

R is methyl or ethyl in the formula I, R ⁴And R ⁵Respectively be hydrogen, or R ⁴And R ⁵Among one be hydrogen, another is (C ₁-C ₆) alkyl or (C ₆-C ₈) methyl cycloalkyl,

This method comprises the following steps:

(a) make R-trans-3-methyl-4-heptenoic acid, R-is trans-and the activated form of 3-ethyl-4-heptenoic acid or S-3-methyl-6-heptenoic acid carries out coupled reaction with the compound with following absolute stereo chemical formula, formation has the compound of absolute stereo chemical formula (III c)

R wherein ⁶And R ⁷Respectively be benzyl, or R ⁶And R ⁷Among one be benzyl, another is (C ₁-C ₆) alkyl or (C ₆-C ₈) methyl cycloalkyl,

In the formula (III c), two keys of any one representative in two dotted lines, but be not that two dotted lines are all represented two keys, collateral condition is when two key positions endways 6,7 o'clock, R is a methyl, and

(b) formula III c compound forms the formula I compound through hydrogenation, two keys is reduced simultaneously and carbobenzoxy group carries out hydrogenolysis.

13, in accordance with the method for claim 12, wherein Suan activated form is an acyl chlorides, R ⁶And R ⁷Preparation all is a benzyl, R ⁴And R ⁵All be hydrogen.

14, the preparation have the absolute stereo chemical formula (III is the method for compound a),

Two keys of the representative of any one in two dotted lines wherein, but be not that two dotted lines are two keys, R is methyl or ethyl, and collateral condition is two keys positions 6,7 o'clock endways, and R is a methyl,

This method comprises the following step:

(a) make R-trans-3-methyl-4-heptenoic acid, R-is trans-activated form of 3-ethyl-4-heptenoic acid or S-3-methyl-6-heptenoic acid with have following absolute stereo chemical formula compound and carry out coupled reaction, form the compound of absolute stereo chemical formula (III b),

R wherein ⁶And R ⁷Each is benzyl naturally, (C ₁-C ₆) alkyl or (C ₆-C ₈) methyl cycloalkyl,

(b) (III is compound a) to make hydrolysis of ester group in formula (III b) compound form formula.

15, in accordance with the method for claim 14, wherein Suan activated form is an acyl chlorides, R ⁶And R is a benzyl.

16, preparation has the method for following absolute stereo chemical formula crystalline compounds, and this method comprises makes the crystallization from one or more organic solvents of this compound.