KR900004106B1 - Method for preparing N- (S-3-alkylheptanoyl) -D-gamma-glutamyl-glycyl-D-alanine and intermediates thereof - Google Patents

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Description

Method for preparing N- (S-3-alkylheptanoyl) -D-gamma-glutamyl-glycyl-D-alanine and intermediates thereof

-3-알킬-4-헵테노산(VIb,하기에 명시),

-3-메틸-6-헵테노산(Xb, 하기에 명시) 및

-3-알킬-헵테노산의 제조방법 및 그 중간체 ; 일반식(I)의 화합물의 합성에서 중간체로서 유용한 절대 입체화학적 일반식(II)의 후자물질의 제조방법 및 그 중간체 ; 및 절대 입체화학적 일반식(III)의 면역조절제(또는 전구체)에 관한 것이다.The present invention provides a useful method for preparing an immunomodulator of general formula (I) below; Trans

3-alkyl-4-heptenoic acid (VIb, specified below),

-3-methyl-6-heptenoic acid (Xb, specified below) and

Method for preparing 3-alkyl-heptenoic acid and intermediates thereof; A method for preparing the latter of the absolute stereochemical formula (II), which is useful as an intermediate in the synthesis of the compound of formula (I), and its intermediates; And immunomodulators (or precursors) of absolute stereochemical formula (III).

Wherein R is methyl or ethyl and R ⁴ and R ⁵ are each hydrogen or one of R ⁴ and R ⁵ is hydrogen and the other is (C ₁ -C ₆ ) alkyl or (C ₆ -C ₈ ) Cycloalkylmethyl, R ² and R ³ are both hydrogen (IIIa) or a pharmaceutically acceptable salt thereof or each independently represent (C ₁ -C ₆ ) alkyl, (C ₆ -C ₈ ) cycloalkylmethyl or benzyl (IIb), one or the other (but not both) of the dashed line represents a double bond, where R is methyl if the double bond is at the terminal 6,7-position.

-3-메틸헵타노산(II, R=CH₃)은 처음에는-15℃에서 퀴닌염의 다중 결정화에 의해 불특정 수율로써 상응하는 라세메이트로부터 제조하였다[참고문헌 : Levene et a1., J.Biol.Chem., 95,pp.1-24, 1932, page 18, 여기서는 2-n-부틸부티르 산-4로 칭한다]. 그후 광학적으로 활성인 3-메틸헵타노산은 수많은 다른 방법에 의해 제조되어 왔으나[참고 : Soai et al., J.Chem.Soc., Chem.Commun.1985,pp.469-470; Oppolzer et al., Helv.Chim.Acta.68,pp.212-215(1985); Ohno et al., U.S.Patent 4,564,620(1986) : Mori et al., Synthesis 1982,pp.752-753; Oppolzer et al., Helv.Chim.Acta.64,pp.2808-2811(1981) ; Mukaiyama et a1., Chem.Lett.1981,pp.913-916 ; Posner et al., J.Am.Chem.Soc.103,pp.2886-2888(1981) : Mukaiyama et al., Bu11.Chem.Soc.Japan,51,pp.3368-3372(1978) : Meyers et al., J.Am.Chem.Soc.98,pp.2290-2294(1976)]. 일반적으로 이들 제조법은 한가지 이상의 단점(최대 가능 수율은 50%이며, 이때 원하지 않는 부산물의 적어도 50% 처분이 필요하며, 원하지 않는 부산물의 적어도 50%은 마지막 제조단계를 통해 처리되며; 생성물 산은 광학적으로 순수하지 못하며 : 유기 금속 시약의 사용은 큰 규모로 취급할 때 어려움이 뒤따르며 : 총 수율이 낮으며/거나 필요한 시약을 쉽게 구할 수 없다.)을 감수해야 한다.Optically pure

3-Methylheptanoic acid (II, R = CH ₃ ) was initially prepared from the corresponding racemates in unspecified yield by multiple crystallization of quinine salts at -15 ° C. See Levene et al., J. Biol. Chem., 95, pp. 1-24, 1932, page 18, here referred to as 2-n-butylbutyric acid-4]. Optically active 3-methylheptanoic acid has since been prepared by a number of different methods [see: Soai et al., J. Chem. Soc., Chem. Comm. 1985, pp. 469-470; Oppolzer et al., Helv. Chim. Acta. 68, pp. 212-215 (1985); Ohno et al., US Pat. No. 4,564,620 (1986): Mori et al., Synthesis 1982, pp. 752-753; Oppolzer et al., Helv. Chim. Acta. 64, pp. 2808-2811 (1981); Mukaiyama et al., Chem. Lett. 1981, pp. 913-916; Posner et al., J. Am. Chem. Soc. 103, pp. 2886-2888 (1981): Mukaiyama et al., Bu11. Chem. Soc. Japan, 51, pp. 3368-3372 (1978): Meyers et al., J. Am. Chem. Soc. 98, pp. 2290-2294 (1976). In general, these recipes have one or more disadvantages (maximum possible yield is 50%, requiring at least 50% disposal of unwanted byproducts, at least 50% of unwanted byproducts being processed through the last preparation step; product acid is optically Not pure: The use of organometallic reagents is difficult when handling on a large scale: low total yield and / or the necessary reagents are not readily available.).

Currently used asymmetric epoxy reactions (called Sharpless decomposition) are described in Katsuki and Sharpless, J. Am. Chem. Soc. 102, pp. 5974-5976,1980; Sharpless et a1., Pure App1. (Chem. 55, pp. 589-604, 1983), the stereospecific Claisen condensation reaction has already been published in US Pat. No. 4,045,475, invented by chan et al.

The relatively new field of immunopharmacology, and especially those treated as immunomodulation, continues to develop rapidly. Various naturally occurring compounds are investigated, including tetrapeptide tuftsin chemically known as N ^2- [1- (N ² -L-threonyl-L-lysyl) -L-prolyl] -L-arginine have. Much attention is focused on synthetic peptidoglycan derivatives, particularly those known as muramyl dipeptides.

The immunomodulators of general formula (I) and their use, generally as amorphous lyophilate when R ⁴ = R ⁵ = H, are pending PCT application No. PCT / US, filed November 25, 1985 Already announced in 85/02351.

Since this application has not yet been published, the preparation of these compounds and their use are cited in the context of the present invention for practical use.

Other immunostimulant peptides have been described in numerous patent specifications: L-alanyl-alpha-glutaric acid N-acyldipeptides are in Federal Republic of Germany No. 3,024,355, issued January 15, 1981; Tetra- and penta-peptides containing either the D-alanine-L-glutamyl component or the L-alanyl-D-glutamyl component are disclosed in British Patent No. 2,053,231, published February 8, 1981, January 8, 1981, respectively. And N-acyl-alanyl-gamma-D-glutamyl tripeptides derivatives wherein the C-terminal amino acid is lysine or diaminopimelic acid, is disclosed in the Federal Republic of Germany, issued January 15, 1981. No. 3,024,369: lactoyl tetrapeptides consisting of the N-lactyalanyl, glutamyl, diaminopimeryl and carboxymethylamino components are in European Patent No. 11283, issued May 13, 1980; US Pat. Nos. 4,311,640 and 4,322,341 for polypeptides having the general formula (A) and those derivatives protecting carboxy and amino groups; European Patent No. 25,482; No. 50,856; No. 51,812; No. 53,388; 55,846 and 57,419;

Wherein R ^a is hydrogen or acyl; R ^b is especially hydrogen, lower alkyl, hydroxymethyl, benzyl; R ^c and R ^d are each hydrogen, carboxy, —CONR ^g R ^h (R ^g is hydrogen, optionally lower alkyl substituted with hydroxy and R ^h is mono-dicarboxy lower alkyl); R ^e is hydrogen or carboxy when one of R ^d and R ^e is hydrogen and the other is carboxy or —CONR ^g R ^h : m is 1 to 3; n is 0 to 2;

Peptides similar to compounds of formula (A) are described in US Pat. No. 4,565,653 and European Patent No. 157,572 when R ⁴ is a basic amino acid component, and European Patent Application No. 178,845 when R ⁴ is a non-uniform amino acid. Described in the heading.

A compound of formula (A) wherein n is 1; R ^a is CH ₃ CH (OH) -CO-; R ^b is CH ₃ ; R ^c and R ^e are each -COOH; R ⁴ is N ² (gamma-D-glutamyl) -meso-2 (L) .2 (D) -diamino-P as a microstructure capable of eliciting the biological response properties of -CONHCH ₂ COOH and R ^f is H Melic acid is published in the following literature (kitaura et al., J. Med. Chem., 25,335-337 (1982)). The compound of formula (A) is known as FK-156.

-3-메틸헵타노일)-D-감마-글루타밀-글리실-D-알라닌의 더욱 안정하고 결정형 형태를 제공한다.The present inventors have now described the immunomodulatory compounds of formula (A) (when R = CH ₃ and R ⁴ = R ⁵ = hydrogen), i.

3-methylheptanoyl) -D-gamma-glutamyl-glycyl-D-alanine provides a more stable and crystalline form.

-3-알킬-4-헵테노산(VIb, 하기에 명시) 및-

-3-알킬헵테노산(II, 상기에 명시)의 효율적인 제조방법을 제공한다.We now also remove organometallic reagents and wastewater by-products of R-enantiomers produced during the second half of the synthesis, with high optical purity intermediates with excellent total yield.

-3-alkyl-4-heptenoic acid (VIb, specified below) and-

Provided is an efficient process for the preparation of 3-alkylheptenoic acid (II, specified above).

As a route, the initial step is the chaliceless degradation of trans-4-alkene-3-ols of the general formulas (IV and V).

Wherein R is as defined above.

-엔안티오머를 산화시키고 절대 입체화학적 일반식(V)의 원하는 미반응

-엔안티오머를 유지시키기에 충분한 양으로 일반식(IV)의 라세메이트를

-부틸 하이드로페록사이드와 반응시키는 것이다.In particular the step is carried out under a reaction inert solvent in the presence of titanium tetraisopropoxide and L-(+)-diisopropyl tartrate,

-Oxidize enantiomers and desired unreacted absolute absolute chemical formula (V)

-Racemate of formula (IV) in an amount sufficient to maintain the enantiomer

It is reacted with -butyl hydroperoxide.

-3-알킬-4-헵테노에이트를 수득한다.In the second step, the compound of general formula (V) is condensed with tri [(C ₁ -C ₃ ) -alkyl] orthoacetate in stereospecificity, and the intermediate allyl-enol ether in the presence of an acid in the reaction inert solvent without separation Rearranged to (C ₁ -C ₃ ) alkyl of the following absolute stereochemical formula (VIa)

Obtain 3-alkyl-4-heptenoate.

(VIa) X = OR ¹ ; R ¹ = (C ₁ -C ₃ ) alkyl

(VIb) X = OH

(VIc) X = Cl

(VId) X = H

-3-알킬-4-헵테노산(VIb)으로 가수분해 시킨 후, 필요하면

-3-알킬헵테노산으로 수소화시 킨다.In one way, trans ester

After hydrolysis with 3-alkyl-4-heptenoic acid (VIb), if necessary

Hydrogenated with 3-alkylheptenoic acid.

-3-알킬헵타노에이트를 수득한 후 가수분해하여 일반식(II)의 산을 얻는다. 또 다른 방법으로는, 일반식(V)의 분해된 트랜스-4-알켄-3-올을 머규릭 아세테이트 [Hg(OAc)₂)] 촉매 존재하에 에틸 비닐 에테르의 작용에 의해 알릴 비닐 에테르(IX)로 전환시킨다. 알릴 비닐 에테르를 가열하여 일반식(VId)의

-3-알킬-4-헵테날로 재배열하고, 이어서 예를 들어 존스(Jones) 시약(희석 무기산 중에 크롬무수물)으로 산화시켜 상기 R-3-알킬-4-헵테노산을 형성한다.Alternatively, the ester is first hydrogenated to (C ₁ -C ₃ ) alkyl

The 3-alkylheptanoate is obtained and then hydrolyzed to give an acid of the general formula (II). Alternatively, the degraded trans-4-alkene-3-ol of general formula (V) may be reacted by the action of ethyl vinyl ether in the presence of a mercury acetate [Hg (OAc) ₂ ) catalyst. Switch to). Allyl vinyl ethers are heated to

Rearrange to -3-alkyl-4-heptenal and then oxidize, for example, with Jones reagent (chromium anhydride in dilute inorganic acid) to form the above R-3-alkyl-4-heptenoic acid.

-시트로넬롤로부터 유기 금속시약 및

-엔안티오머의 페수성 부산물로 회피하면서, 우수한 총 수율로써 고 광학순도의 중간체

-3-메틸-6-헵테노산(Xd, 하기에 명시) 및

-3-메틸헵타노산(II, R=CH₃, 상기에 명시)의 유용한 제조방법을 제공한다.In addition, the inventors have found that the absolute stereochemical formula (XI)

Organometallic reagents from citronellol and

Intermediates of high optical purity with good total yield, avoiding with the water-soluble by-products of enantiomers

-3-methyl-6-heptenoic acid (Xd, specified below) and

Provided are useful methods for the preparation of 3-methylheptanoic acid (II, R = CH ₃ , specified above).

The initial stage of this synthesis is the optional protection of alcohol groups with conventional and conventional silylprotecting groups (such as t-butyldimethylsilyl groups), followed by ozone decomposing by methylsulfido action to yield the following absolute stereochemical formula To yield a new compound of XII):

-시트로넬롤로 예전에 사용된 방법과 유사하다.Wherein R ⁸ is hydrogen (XIIa) or silyl hydroxyprotecting group (XIIb). The method used is specifically shown in the following preparation to prepare R-enantiomers of compounds of formula (V) wherein R ⁸ is t-butyldimethylsilyl.

It is similar to the method previously used for citronellol.

Although not used here, the latter is used for the synthesis of proxyformin (Tapolczay et al., J. Chem. Soc., Chem. Commun. 1985, pp. 143-145).

-3-메틸-6-헵테노산을 형성하고 ; 필요하면, (d) 일반식(Xd)의 화합물의 촉매 수소화 반응하여 일반식(II) (여기서 R은 메틸이다.)의

-3-메틸헵타노산을 형성하는 단계를 포함한다.The invention particularly relates to the conversion of intermediate compounds of the general formula (V) and the following general formula (X) and compounds of the general formula (XII) to optically active acids of the general formulas (Xd) and (II, R = CH ₃ ). And (a) a compound of formula (XII) is reacted with methylenetriphenylphosphorane [CH ₂ = P (C ₆ H ₅ ) ₃ ] to form a compound of formula (Xa) or (Xb). To form; (b) when the compound is of formula (Xa) to form a compound of formula (Xb), dilute inorganic acid hydrolysis is carried out as a separate step or simultaneously with the subsequent step; And (c) oxidizing the compound of general formula (Xb) with chromium anhydride in dilute inorganic acid to

To form 3-methyl-6-heptenoic acid; If necessary, (d) catalytic hydrogenation of the compound of general formula (Xd) to yield a compound of general formula (II), wherein R is methyl.

Forming 3-methylheptanoic acid.

(Xa) R ⁹ = CH ₂ OR ¹⁰ , R ¹⁰ = silyl protecting group

(Xb) R ⁹ = CH ₂ OH

(Xc) R ⁹ = CHO

(Xd) R ⁹ = COOH

(Xe) R ⁹ = COCl

-t-부틸펜에틸디메틸실릴 및 t-부틸디메틸실릴이다. 바람직한 희석 무기산은 H₂SO₄이다. 이것은

-시트로넬롤을 메틸설파이드 작용으로 가온존 분해에 의해 일반식(XIIa)의 화합물 : 및 (e)

-시트로넬롤을

-부틸디메틸실릴 클로라이드와 반응시키고; 및 (f) 메틸설파이드 작용으로 얻어진 하이드록시 보호 시트로넬롤을 가오존 분해하는 추가의 단계에 의해 R⁸이 t-부틸디메틸실릴 일때 일반식(XIIb)의 화합물을 합성하는 것이 바람직하다.As hydrogen R ⁸ are a few steps in the glass but, as the silyl protecting group R ⁸ is advantageous in the purification process easier to separate the intermediate product from step (a). Preferred hydroxy protecting groups are trimethylsilyl,

-t-butylphenethyldimethylsilyl and t-butyldimethylsilyl. Preferred dilute inorganic acids are H ₂ SO ₄ . this is

-A compound of general formula (XIIa) by warm zone decomposition of citronellol by methylsulfide action: and (e)

-Citronellol

Reacted with -butyldimethylsilyl chloride; And (f) a further step of ozone degrading the hydroxy protective citronellol obtained by the methylsulfide action, to synthesize the compound of formula (XIIb) when R ⁸ is t-butyldimethylsilyl.

-3-알킬-4-헵테노산 또는

-3-메틸-6-헵테노산(예를 들어, 산염화물이 VIC 또는 Xe일 때)의 활성화된 형태를 하기 일반식(Ⅶ)의 화합물과 커플링하고; (b) 수소화 반응 촉매 존재중의 반응 불활성 용매중에서 이중 결합의 동시 감소와 함께 상기 중간체 화합물의 수소화 반응 및 벤질 그룹 또는 그룹들의 수소화 가수분해하는 단계를 포함하는 일반식(I)의 면역 조절제의 개선된 제조방법에 관한 것이다;The present invention also provides a process for preparing an intermediate compound of formula (IIIb) wherein R ² and R ³ correspond to R ⁶ and R ⁷

-3-alkyl-4-heptenoic acid or

Coupling an activated form of -3-methyl-6-heptenoic acid (eg, when the acid chloride is VIC or Xe) with a compound of formula (VII); (b) improving the immunomodulator of formula (I) comprising the hydrogenation of the intermediate compound and the hydrolysis of the benzyl group or groups with simultaneous reduction of double bonds in a reaction inert solvent in the presence of a hydrogenation catalyst To a method of preparation;

Wherein R ⁶ and R ⁷ are both benzyl, or one of R ⁶ and R ⁷ is benzyl and the other is (C ₁ -C ₆ ) alkyl or (C ₆ -C ₈ ) cycloalkylmethyl.

Finally, the present invention relates to immunomodulators and / or precursors of compound (I) of general formula (III). Compounds of formula (IIIa) (ie R ² = R ³ = hydrogen) are obtained by conventional ester hydrolysis of diester compounds of formula (IIIb).

The expression "reactive inert solvent" as used herein means that it does not interact with the starting materials, intermediates or products in a way that adversely affects the yield of the desired product.

Preferred material of R ¹ is ethyl. Preferred acid catalysts are Lewis acids (eg dry HCI, AlCl ₃ ). While not intending to limit the invention, preferred raw materials of racemic trans-4-hexene-3-ol or trans-4-hepten-3-ol are selected from the group consisting of ethyl grignard reagent and crotonaldehyde or 2-pentenal. And through each reaction.

-트랜스-4-알킬-3-올(V)을 비대칭 에폭시 반응, 소위 샤플리스 분해(상기에서 인용된 문헌 참조)의 방법에 의해 얻는다. 본 방법에 따라, 원하지 않는 라세메이트(IV) 중의

-트랜스-2-헥센-4-올을 L-(+)디이소프로필 타르트레이트의 과량 몰 존재하에서 선택적으로 t-부틸하이드로페록사이드(적어도 0.5몰, 단 라세믹 헥센올 몰당 1몰 미만으로 한정)와 반응시키고 이어서 티타늄테트라이소프로폭사이드의 1몰 당량과 반응시킨다. 반응을 일반적으로-20 내지-80℃범위의 냉각 온도, 메틸렌클로라이드와 같은 반응 불활성 용매중의 무수 조건하에서 수행한다. 원하는 미반응

-트랜스-2-알켄-4-올(V)을 중류 또는 크로마토그라피와 같은 표준 방법으로 분리시킨다. 원하는 생성물의 상대적으로 높은 휘발성 때문에, 상기 크로마토그라피에서는 펜탄 및 에테르와 같은 저비점 용매를 사용하는 것이 바람직하다.The invention is easily carried out.

-Trans-4-alkyl-3-ol (V) is obtained by the method of asymmetric epoxy reactions, the so-called Shapless decomposition (see literature cited above). According to the method, in unwanted racemate (IV)

-Trans-2-hexene-4-ol is optionally limited to t-butylhydroperoxide (at least 0.5 mole, but less than 1 mole per mole racemic hexenol, in the presence of excess mole of L-(+) diisopropyl tartrate) ) Followed by 1 molar equivalent of titanium tetraisopropoxide. The reaction is generally carried out under anhydrous conditions in a reaction inert solvent such as methylene chloride, at a cooling temperature ranging from -20 to -80 ° C. Desired unreacted

-Trans-2-alkene-4-ol (V) is separated by standard methods such as midstream or chromatography. Because of the relatively high volatility of the desired product, it is preferable to use low boiling point solvents such as pentane and ether in the chromatography.

-트랜스-4-알켄-3-올(V)과 (C₁-C₃)트리알킬 오르토아세테이트의 축합/클래센 재배열이다. 반응을 반응 불활성 용매하에서 수행할 수 있지만, 일반적으로 승온(예를 들어,120 내지 160℃)에서 오르토아세테이트 에스테르의 과량하에서 반응을 간략하게 수행하는 것이 바람직하다. 트리에틸 오르토아세테이트(비점 142℃)가 시약일 때, 환류 온도를 사용하는 것이 통상적이다. 반응을 산 촉매의 존재하에서 수행한다. 프로피온 산, 피발산, 2,4-디니트로페놀, 건조 HCl 및 AlCl₃와 같은 산이 효과적이다. 바람직한 촉매는 루이스 산이고 가장 바람직한 것은 AlCl₃이다.The next step is

- trans-4-alkenyl-3-ol (V) and (C ₁ -C ₃₎ a condensation / rearrangement metallocene class of trialkyl orthoacetate. The reaction can be carried out in a reaction inert solvent, but it is generally preferred to carry out the reaction briefly in excess of orthoacetate ester at elevated temperature (eg 120-160 ° C.). When triethyl orthoacetate (boiling point 142 ° C.) is a reagent, it is common to use a reflux temperature. The reaction is carried out in the presence of an acid catalyst. Acids such as propionic acid, pivalic acid, 2,4-dinitrophenol, dry HCl and AlCl ₃ are effective. Preferred catalyst is Lewis acid and most preferred is AlCl ₃ .

Alternatively, the obtained ester of formula (VIa) is hydrolyzed by conventional methods to obtain the corresponding unsaturated acid of formula (VIb).

-시트로넬롤(XI)을 처음에 통상적인 방법에 의해 일반식(XII)의 광학적 활성 출발물질중의 하나로 전환시킨다(상기에 명시됐고 하기 제조예에 예시됨). 화합물(XII)을 상기 일반식(Xd)의

-3-메틸-6-헵테노산 또는

-3-메틸헵-타노 산으로 각각 전환시킨다(단계적 유형은 다음 분맥에서 상세히 서술한다).Also other methods of the present invention are readily performed. Commercially available

Citronellol (XI) is first converted to one of the optically active starting materials of the general formula (XII) by conventional methods (specified above and illustrated in the preparation below). Compound (XII) is substituted with the general formula (Xd)

-3-methyl-6-heptenoic acid or

Convert to 3-methylhep-tano acid, respectively (step type is detailed in the next branch).

In the first step, hydroxy or hydroxy protecting aldehyde (XIIa or XIIb) is a reaction inert aprotic solvent, for example a mixture of tetrahydrofuran and hexane, for example (methyl) triphenylphosphonium halide ( Usually, a wittig reaction is performed from bromide) and butyllithium to methylenetriphenylphosphorane, which is generally formed in situ.

If temperature is not an important factor for phosphoran formation, the preferred temperature is in the range of ± 25 ° C, most preferred in the range of ± 10 ° C. Aldehyde (XIIa) or hydroxy protecting aldehyde (XIIb) is reacted with phosphorane in similar solvents and similar temperature ranges to form hydroxy or hydroxy protecting olefins of formula (Xb) or (Xa), respectively.

In the second step, when only hydroxy protecting groups are required to be present, the protecting groups are most commonly removed by hydrolysis by the aqueous acid conditions used in the third step. This third step is the (Xd) Jones oxidation of the primary alcohol (Xb) with acid. The Jones oxidation reaction uses an aqueous solution of H ₂ CrO ₄ formed from the so-called Jones reagent, CrO ₃ and strong acid. Typically, the Jones reagent is prepared from an excess of concentrated H ₂ SO ₄ and CrO ₃ with a weight of about 1: 1 water and then diluted with water to the desired concentration, for example about 3M. Alcohol (XIIa) The protected alcohol (XIIb) is generally reacted with at least 2 molar equivalents of Jones reagent in a solution of soluble water and a reaction inert organic solvent such as acetone. Under these conditions, alcohols are formed by rapid hydrolysis of certain silyl ether groups, followed by oxidation with acid (Xd). Temperature is not an important factor, for example 0 to 50 ° C. is usually satisfactory, and circulating temperature, for example 17 to 27 ° C. is most convenient.

The initial oxidation of the alcohol results in a Jones oxidation reaction to form an aldehyde of the general formula (Xc), but usually not separated in the Jones oxidation reaction. If separation of the intermediate aldehyde is desired, the alcohol is oxidized with a more selective oxidant, such as pyridinium chloromate, which obviously yields the intermediate aldehyde (Xc), followed by oxidation with acid (Xd) using Jones reagents or other suitable reagents.

If necessary, the active form (for example, as acid chloride of formula (VIc) or (Xe), as a conventional mixed anhydride, by activation with a conventional dehydrating binder such as dicyclohexylcarbodiimide) is conventional The method is coupled with a compound of formula (VIII) to form an immunomodulatory diester compound of formula (IIIb).

Wherein R ⁹ and R ¹⁰ are each independently (C ₁ -C ₆ ) alkyl, (C ₆ -C ₈ ) cycloalkylmethyl or benzyl. The latter is hydrolyzed by conventional methods to an immunomodulatory dieside compound of formula (IIIa) or a pharmaceutically acceptable salt thereof.

Alternatively, the compound of formula (IIIb) (when R ² and / or R ³ is benzyl) is hydrogenated to form an immunomodulatory compound of formula (I). In the hydrogenation reaction, the double bond is saturated as hydrogen and the benzyl group (s) are hydrogen hydrolyzed. Hydrogenation reactions include hydrogenation catalysts such as nickel or precious metals; The reaction is carried out in a reaction inert solvent on a supported catalyst (eg Raney nickel, pd / c) or an unsupported catalyst (eg RhCl ₃ ). Suitable solvents are not intended to be limited to lower alcohols, but are ethers such as dioxane, detrahydrofuran or diethoxyethane, and esters such as ethyl acetate. Although exothermic to some extent, it is desirable to use a circulating temperature to avoid heating or cooling costs without cooling. Pressure is not an important factor and it is desirable to avoid expensive high pressure equipment as it is below 7 atm. Hydrogenation on Pd / C at 3 to 6 atmospheres is particularly suitable for this modification.

-3-알킬헵타노산(II)을 수득하고, 또한 수소화반응/가수분해 단계를 바꿔(C₁-C₃)알킬

-3-알킬헵타노에이트를 통해 (C₁-C₃)알킬

-

-3-알킬-4-헵테노에이트(V I a)로부터 얻는다. 마지막으로 S-3-알킬헵타노산을 상기에 상세하게 서술한 방법으로 활성화시키고, 상기 일반식(V-II)의 디에스테르와 커플링하고 상술된 방법에 의해 상기 일반식(I)의 면역조절 화합물을 형성시킨다.Alternatively, the unsaturated acid of formula (VIb) or (Xd) may be hydrogenated under the same conditions as detailed in the preceding context.

Obtain 3-alkylheptanoic acid (II), and also change the hydrogenation / hydrolysis step (C ₁ -C ₃ ) alkyl

(C ₁ -C ₃ ) alkyl through -3-alkylheptanoate

-

Obtained from 3-alkyl-4-heptenoate (VI a). Finally, S-3-alkylheptanoic acid is activated in the manner described in detail above, coupled with the diester of formula (V-II) and immunomodulation of formula (I) by the method described above To form a compound.

-4-알켄-3-올(V)을 25 내지 40℃ 온도 범위에서, 통상적으로 에틸 비닐 에테르 (비점 36℃)의 환류 온도에서 반응 활성 용매(바람직하게는 에틸비닐 에테르의 과량)중의 촉매로서 Hg(OAc)₂존재하에서 에틸 비닐 에테르의 작용에 의해 일반식 (IX)의 상응 비닐 에테르로 전환시킨다. 얻어진 비닐 에테르(IX)를 일반적으로 크실렌 또는 데칼린과 같은 고비점, 친액성 반응불활성 용매하에서, 이것을 일반식(VId)의 불포화 알데하이드로 입체 특이성 재배열을 위한 필요한 압력하에서 140 내지 200℃로 가열시킨다. 일반식(VIb)의 불포화 산을 얻기 위해, 알데하이드를 CrO₃와 강산으로 부터 형성된 H₂CrO₄의 수성액, 즉 소위 통상적인 존스 시약으로써 쉽게 산화시킨다. 전형적으로 존스시약은 진한 H₂SO₄및 CrO₃의 과량과 함께 약 1 : 1 물 중량으로부터 제조된 후, 원하는 농도, 예를들어약 3M 로 물로써 희석시킨다. 일반적으로 용해성 물중의 용액에서 불포화 산(V Ib), 불포화 알데하이드(VId)를 형성하기 위해, 아세톤 같은 반응 불활성 유기 용매를 적어도 1몰 당량의 존스 시약과 반응시킨다. 온도는 중요한 요인이 아니며, 예를들어 0 내지 50℃가 보통 만족스럽고 순환 온도, 예를들어 17 내지27℃가 가장 만족스럽다.In another method of the present invention,

4-alkene-3-ol (V) as a catalyst in the reaction active solvent (preferably excess of ethylvinyl ether) at a reflux temperature of 25 to 40 ° C., typically at ethyl reflux (boiling point 36 ° C.) Conversion to the corresponding vinyl ether of formula (IX) by the action of ethyl vinyl ether in the presence of Hg (OAc) ₂ . The vinyl ether (IX) obtained is generally heated to 140-200 ° C. under high pressure, lipophilic reactive inert solvents such as xylene or decalin, under the necessary pressure for stereoaldehyde specific rearrangement of unsaturated aldehydes of formula (VId). . To obtain the unsaturated acid of formula VIb, the aldehyde is easily oxidized with an aqueous solution of H ₂ CrO ₄ formed from CrO ₃ and a strong acid, ie the so-called conventional Jones reagent. Jones reagents are typically prepared from about 1: 1 water weight with excess H ₂ SO ₄ and CrO ₃ and then diluted with water to the desired concentration, for example about 3M. Generally, a reaction inert organic solvent such as acetone is reacted with at least one molar equivalent of Jones reagent to form unsaturated acids (V Ib), unsaturated aldehydes (VId) in a solution in soluble water. Temperature is not an important factor, for example 0 to 50 ° C. is usually satisfactory and circulating temperature, for example 17 to 27 ° C. is most satisfactory.

Crystalline formation of the compound of formula (I) (when R is methyl and R ₄ = R ₅ = hydrogen) is obtained by crystallization from an organic solvent or a composite of organic solvents. Suitable solvents are acetone, acetonitrile / ethanol and tetrahydrofuran / ether. Preferred solvents in terms of product recovery are acetonitrile / ethanol, but acetone is preferred in terms of product purity. This novel form has a clear stability advantage at the previous stage of amorphosthiophylate. It is more dense and less electrostatic, making it very easy to handle in the manufacture of more delicate dosage forms.

The pharmaceutically acceptable mono- and dibasic salts of the compounds of formula (I) or (IIIa) generally have a suitable conformation as a base such as NaOH, KOH, Na ₂ CO ₃ or an amine in solution, preferably an aqueous solution of acid. Obtained by treatment in chemical proportions. Salts are separated by evaporation or precipitation.

The product of formula (I) or (III) of the present invention is a pharmaceutical agent in mammals, including humans, for the clinical and therapeutic treatment of diseases caused by various pathogenic microorganisms, in particular gram-negative bacteria Useful as They are also useful as immune stimulants in mammals, including humans, with an increased risk of infection due to latent or clinically-induced immune abolition. The test procedure uses normal or immune C ₃ H / HeN male mice obtained from the Charles River Breeding laboratory. Adapt for 5 days before using mice and subcutaneously (sc) or orally (0.2) with various dilutions (100, 10, 1 and 0.1 mg / kg) of test compound or placebo (pyrogen free saline) using 0.2 ml amounts. po). Treated herds depend on the infectious organism used: pneumococcal inoculated pneumonia 24 and 0 h before immunization: E. coli or staphylococci in immunized mice 3, 2 and 1 day prior. The dose is administered intramuscularly for pneumonia and intraperitoneally for E. coli or staphylococci. Use 0.2 ml amount for dosing. Survival water is recorded after 7 days for pneumonia and 3 days after the other two microbial inputs.

Culture Preparation: Pneumococcal, Escherichia Coli or Staphylococcus: Streaking the culture for purity from frozen blood storage on brain heart dehumidifier (BHl) agar. Three cells are extracted from an 18 hour plate incubation and placed in 9 ml BHI broth. After streaking 0.2 ml on several BHI agar slant surfaces, the broth is incubated for 2 hours at 37 ° C. on a rotary stirrer. Following 18 hours of incubation at 37 ° C., the slant is washed with BHI broth, followed by appropriate dilution to adjust the culture concentration using SPECTRONIC 20 and to reach the LD90 input level in normal mice.

When used as an anti-infective or immunostimulating agent in humans, the compounds (I) or (III) of the present invention are generally in the form of compositions that are standard in pharmaceutical practice, via the intramuscular, intramuscular, intravenous or intraperitoneal route. It is usually administered. For example, they may be administered in the form of tablets, pills, powders or granules containing excipients such as starch, milk sucrose, specific forms of clay and the like. They can be administered encapsulated in admixture with the same or equivalent excipients. They may also be administered in the form of elixirs, which may contain caliber suspensions, solutions, emulsions, syrups and sweeteners and colorants. In the case of caliber administration as a therapeutic agent of the invention, tablets or capsules containing about 50 to about 500 mg of the active ingredient are most suitable for application.

The physician will determine the most appropriate dosage for each patient and age. It will vary according to body weight and specific patient response and route of administration. Preferred aperture dosage ranges are from about 1.0 to about 300 mg / kg / day in single or divided doses: the most preferred range is about 1.0 to about 20 mg / kg / day.

The following examples are presented by way of illustration and are not intended to limit the invention, and many variations of the invention are possible within the spirit and scope of the invention.

Example 1

R-trans-4-hexene-3-ol

Into a 500 ml three neck round bottom flask equipped with a magnetic stirrer, septum, thermometer and N ₂ inlet are placed 270 ml CH ₂ Cl ₂ and 7.2 g of 4A form molecular sieve. Ti [OCH (CH ₃ )] (10.7 ml, 0.036 mol) was added via syringe and the mixture was cooled to -66 ° C and diluted with 10 mlCH ₂ Cl ₂ to reduce viscosity at this temperature. Diisopropyl tartrate (10.lg, 0.043 moles) is added via cannula and washed with 5 ml additional CH ₂ Cl ₂ . After raising the temperature to -62 ° C, racemic trans-2-hexene-4-ol (3.60 g, 0.036 mol) is added at this temperature and washed with 5 ml, CH ₂ Cl ₂ . After stirring for 8 minutes in an acetone / dry ice bath, the temperature is lowered to -68 ° C, at which time t-butyl hydroxide (7.18 ml of 3M toluene, 0.022 mol) is added via syringe. The reaction mixture is warmed to −35 ° C. and held at this temperature in the freezer for 18 hours. The cooling mixture is then filtered using a f1uted filter paper and then mixed into a stirred mixture of 540 ml acetone and 11 ml H ₂ O cooled to -20 ° C. The mixture is stirred for 20 hours while slowly heating to room temperature. The mixture is filtered over CH ₂ Cl ₂ wash over diatomaceous earth. The synthesized filtrate and wash are stripped as oil (17.62 g) without heating. The top stripping material is avoided because the desired product is very volatile. The oil was chromatographed on 176 g silica gel using 4: 1 hexanes: isopropyl ether as eluant and identified by TLC.

The solvent is removed by distillation using a small column packed with glass helice equipped with various take-off heads. The solvent boiling at the upper temperature of 69.5 ° C. (the upper temperature starts to drop at this temperature) is removed as a pot residue, leaving a considerable amount of pure title product as: tlc Rf 0.25 (3: 1 hexanes: ether). The experiment was repeated, using a more volatile 4: 1 pentane: ether as eluent to minimize solvent loss and promote solvent removal and stripping.

Example 2

Ethyl R-3-methyl-4-hexenoate

The product of Example 1 (0.4 g), triethyl-orthoacetate (3 ml) and pivalic acid were heated to reflux for 2.5 hours and cooled to chromatograph the total reaction mixture twice on silica gel, initially as eluent 6 0.45 g of product is obtained using hexane: ether of 1: 1, and then 0.35 g of pure title product is obtained using hexane: ether of 7: 1 as eluent: IR (film) 2960, 1736, 1456, l367, 1334, 1280, 1232, 1172, 1028, 962, 840 cm ^-1

Alternatively, a pentane / ether column fraction containing pure S-trans-2-hexen-4-ol is introduced directly into this step together with pentane and ether initially distilled from the reaction mixture.

Substantially the same results can be obtained by using propionic acid instead of pivalic acid. Dry acid HCl and 2,4-dinitrophenol can also be used successively as acid catalysts, but AlCl ₃ used in the following examples is the preferred catalyst.

Example 3

R-3-methyl-4-heptenoic acid

[Method A]

The product of Example 2 (1.5 g, 0.015 mol) was placed in a 35 ml flask equipped with a magnetic stirrer and a soxhlet extractor containing 4A-type molecular sieves in a thimble. Triethyl orthoacetate (9 ml, (0.49 mol) and AlCl ₃ (0.12 g, 0.009 mol) were then added and the mixture was refluxed on a Soxhlet extractor for 2 hours, yielding the intermediate ethyl R-3-methyl- within this time. TLC confirms the completion of the conversion to 4-hexenoate: TLC-Rf 0.75 (4: 1 hexane: ethyl acetate), Rf 0.85 (15: 5: 2 hexane: ether: acetic acid).

The reaction mixture was cooled down, diluted with ₁₅ ml 2N NaOH and 12 ml CH ₃ OH and stirred at circulating temperature for 27 hours, confirming by TLC that the hydrolysis was complete within this time. The reaction mixture is stripped with methanol, diluted with 12 ml H ₂ O and extracted twice with 25 ml CH ₂ Cl ₂ . The synthesized CH ₂ Cl ₂ is washed back once with 25 ml 2N NaOH. The aqueous layer and countercurrent wash are synthesized, adjusted with concentrated HCl to pH 1, and extracted three times with CH ₂ Cl ₂ . The latter organic layer was synthesized, dried over MgSO ₄ and stripped to give the title product as 1.28 g (60%) oil: tlc Rf 0.65 (15: 5: 2 hexanes: ether: acetic acid): ¹ H-nmr (CDCl ₃ ) delta (ppm) 9.4 (s, 1H), -CO ₂ H), 5.5 (m, 2H, -CH = CH-), 3.0-1.6 (m, 5H), 1.3-0.8 (m, 6H); ir (film) 3400-2400, -2960, 2925, 2860, l708, 1458, 1410, 1380, 1295, 1228, 1190, 1152, 1100, 930 cm ^-1

[Method B]

The ester product of Example 3 (0.24 g) is synthesized with 25 ml CH ₃ OH and 11.5 ml 1N NaOH. The mixture was stirred at circulating temperature for 3.5 hours, and TLC confirmed that hydrolysis was complete within this time. The mixture is extracted twice with 35 ml ether, adjusted to pH 2 with concentrated HCl and extracted three times with 35 ml ether. The acid extract is synthesized, dried and stripped to yield 0.20 g of the same title product as the product of Method A.

Example 4

Ethyl S-3-methylheptanoate

The product of Example 2 (0.20 g) in 40 ml ethyl acetate in a Parr hydrogenation apparatus. Hydrogenation over 0.20 g of 5% Pd / C (50% wettability) for 3 hours under 4 atmospheres of hydrogen. The catalyst is recovered by filtration on diatomaceous earth. Strip the extract of solvent to recover the title product.

Example 5

S-3-methylheptanoic acid

[Method A]

), 1.0(d, -

), 0.6-2.8(m, 잔기 13H) ; ir(필름) 3400-2400, 2960, 2925, 2860, 1708, 1458, 1410, -1380, 1295, 1228, 1190, 1152, 1100, 930cm^-1;

=6.41。(=1% CH₃OH에서) ;

The product of Example 3 (0.20 g) in 40 ml ethyl acetate is hydrogenated in 0.2 g of 5% Pd / C (50% wettability) for 3 hours under 4 atmospheres of hydrogen in a Parr hydrogenation apparatus. The catalyst is recovered by filtration over diatomaceous earth and the extract is stripped to recover the title product as 0.2 g of oil. If necessary, distillation under high vacuum gives the title product: boiling point 77-79 ° C./0.2 mm; ¹ H-nmr (CDCl ₃ ) Delta (ppm): 12.0 (s, -COO

), 1.0 (d,-

), 0.6-2.8 (m, residue 13H); ir (film) 3400-2400, 2960, 2925, 2860, 1708, 1458, 1410, -1380, 1295, 1228, 1190, 1152, 1100, 930 cm ^-1 ;

= 6.41。 (at = 1% CH ₃ OH);

[Method B]

The product of Example 4 is hydrolyzed according to Method B of Example 3 to afford the title product.

Example 6

S-3-methylheptanoyl chloride

The acid product (8.5 g, 0.062 moles) obtained from Example 5 or Example 27 is dissolved with ₁₈ ml CH ₂ Cl ₂ . Oxalyl chloride (5.36 ml, 7.80 g, 0.0614 mole) was mixed into the solution and the mixture was left for 4 hours, demonstrating no further gas evolution within this time. The acid chloride solution is used directly in Example 8, Method C immediately. Alternatively, for use in Example 8, Method C, the acid chlorides are separated from the solvent by stripping and, if necessary, further purified by distillation (boiling point 45 ° C./1.5 mm).

Example 7

R-3-ethyl-4-heptanoyl chloride

The acid product of Example 3 (0.747 g, 5 moles) was converted directly to a CH ₂ Cl ₂ solution of the title product by the method of the above example and used directly in Example 9 below. Alternatively, the reaction mixture is stripped to afford the title product, if necessary distilled under reduced pressure.

Example 8

-3-메틸헵타노일)-D-감마-글루타밀(알파벤질 에스테르)-글리실-D-알라닌 벤질 에스테르-N- (

3-Methylheptanoyl) -D-gamma-glutamyl (alphabenzyl ester) -glycyl-D-alanine benzyl ester-

[Method A]

-3-메틸헵타노일 클로라이드를 가하고 반응 혼합물을 상온에서 80시간 동안 교반시킨다. 메틸렌 클로라이드를 진공하에서 증발시키고 잔유물을 에틸 아세테이트에 용해시킨다. 얻어진 용액을 2.5% 염산, 물, 10% 탄산칼륨, 물 및 불소 용액 순서대로 세척시킨다. 유기상을 분리하고 황산마그네슘상에 전조시키고 진공하에서 농축시킨다. 잔유물을 디에틸 에테르와 함께 가루로 만들고 질소하에서 여과하여 표제 생성물을 수득하며, 이 생성물 전부를 실시예, 방법 A에 직접적으로 사용한다.660 mg (2.0 mg mmol) in a solution of 1.0 g (2.03 mmol) D-gamma-glutamyl (alpha benzyl ester) -glycyl-D-alanine benzyl ester hydrogen chloride (Preparation 4) and 616 mg (6.09 mmol) triethylamine in 50 ml methylene chloride 4.06 mmol)

3-Methylheptanoyl chloride is added and the reaction mixture is stirred at room temperature for 80 hours. Methylene chloride is evaporated in vacuo and the residue is dissolved in ethyl acetate. The resulting solution is washed in the order of 2.5% hydrochloric acid, water, 10% potassium carbonate, water and fluorine solution. The organic phase is separated, precursord to magnesium sulphate and concentrated in vacuo. The residue is triturated with diethyl ether and filtered under nitrogen to afford the title product, which is all used directly in Example, Method A.

[Method B]

The product of Preparation Example 4 (0.75 g, 1.53 mmol), 5 ml CH ₂ Cl ₂ and triethylamine (0.212 ml, 1.53 mmol) were synthesized and stirred under N ₂ . Then scan-3-methylheptanoic acid (Example 5; 0.20 g, 1.39 mmol) and dicyclohexylcarbodiimide (0.286 g, 1.37 mmol) in 4 ml CH ₂ Cl ₂ are added and the reaction is stirred for 16 hours. . The reaction mixture was filtered, the filtrate was stripped to take the residue in 10 ml ethyl acetate and the solution was washed sequentially with 5 ml 2.5% HCl, 5 ml H ₂ O, 5 ml 10% K ₂ CO ₃ and 5 ml fluorine and on MgSO ₄ Drying affords 71 mg (88%) of the title product.

[Method C]

-3-메틸헵타노일 클로라이드의 전량을 온도가 21℃까지 상승할 때까지 15분 동안 가한다. 얼음-물 수조내의 교반을 45분 동안 계속하고, 이 시간내에 겔화성 혼합물이 너무 혼탁하여 더 이상 교반할 수 없게 된다. 겔화성 물질을 쪼개어 125ml의 10% HCl 및 5ml CH₂Cl₂와 혼합시킨다. 유기층을 분리하고, 순서대로 125ml H₂O로 2회, 125ml 10% K₂CO₃로 2회 및 125ml H₂O로 1회 세척하고 MgSO₄상에서 건조시키고 스트립핑하여 82.3g의 습윤성 백색 고체를 얻는다. 500ml의 뜨거운 에틸 아세테이트 내에서 상기 고체를 취한다. 순환온도로 서냉하면서, 표제 생성물을 짙게 결정화하고, 혼합물을 추가의 40ml에틸 아세테이트로서 희석시켜 용이한 교반을 유지시킨다. 정제 표제 생성물을 여과에 의해 회수하고 40℃에서 진공 건조시켜 31.1g생성물(90.5%)을 수득한다.In a 500 ml four-neck round bottom flask equipped with a stirrer, thermometer, dropping funnel and N ₂ inlet, the product of Preparation 4 (32.8 g, 0.059 mol) was dissolved in 175 ml CH ₂ Cl ₂ and cooled to 0-5 ° C. While maintaining the temperature range, triethylamine (24.7 ml, 17.9 g, 0.177 mol, 3 equiv) is added as a slow stream for 15 minutes. In CH ₂ Cl ₂ obtained from Example 6 while maintaining an ice-water bath

The total amount of -3-methylheptanoyl chloride is added for 15 minutes until the temperature rises to 21 ° C. Stirring in the ice-water bath is continued for 45 minutes, within which time the gelable mixture becomes too cloudy and can no longer be stirred. The gelling material is cleaved and mixed with 125 ml of 10% HCl and 5 ml CH ₂ Cl ₂ . The organic layer was separated, washed in sequence twice with _{125ml H 2 O, 125ml 10%} K 2 washed once with twice and 125ml H ₂ O to CO ₃ and dried over MgSO ₄ and stripped to 82.3g of a white solid wetting Get Take the solid in 500 ml of hot ethyl acetate. With slow cooling to circulating temperature, the title product is deeply crystallized and the mixture is diluted with additional 40 ml ethyl acetate to maintain easy stirring. Purification The title product is recovered by filtration and dried in vacuo at 40 ° C. to give 31.1 g product (90.5%).

¹ H-nmr (CDCl ₃ ) delta (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

-3-메틸-4-헵테노일)-D-감마-글루타밀-(알파 벤질 에스테르)-글리실-D-알라닌 벤질-에스테르N- (

3-Methyl-4-heptenoyl) -D-gamma-glutamyl- (alpha benzyl ester) -glycyl-D-alanine benzyl-ester

By the method C of Example 8, the product of Preparation 4 (2.77 g. 5 mmol) is coupled with the total amount of acid chloride in CH ₂ Cl ₂ obtained from Example 7. The washed and dried organic layer is stripped and the product is recovered by dissolving the residue in ethyl acetate and restriping.

Example 10

-3-에틸헵타노일)-D-감마-글루타밀-글리실-D-알라닌-N- (

-3-ethylheptanoyl) -D-gamma-glutamyl-glycyl-D-alanine-

[Method A]

The entire product obtained in Method A of Example 8 is dissolved in 65 ml methanol. Palladium hydroxide (250 mg) is added to the solution and the mixture is stirred under tetrahydrogen atmosphere for 3 hours. The catalyst is filtered off and the solvent is removed in vacuo. The residue is dissolved in water and lyophilized to give the desired product. NMR spectrum (DMSO-d ₆ ) shows absorbency as follows:

8.27-8.03 (m, 3H), 4.32-4.1 (m, 2H), 3.72 (d, J = 6 Hz, 2H), 2.22 (t, J = 8 Hz, 2H), 2.27-1.68 (m, 6H), 1.42 -1.0 (m, 10H), 0.94-0.8 (m, 6H).

When carrying out the weight of the title product of Example 8 (0.50 g) in 25 ml CH ₃ OH using 90 mg 20% Pd (OH) ₂ / C (31% wettability) the method is identical and fuzzy Obtain the electrostatic title product: IR (nujol mull) 3300, 2940, 1740, 1650, 1540, 1468 and 1380 cm ^-1 However, the last two peaks are wide and poorly dissolved.

[Method B]

The product of Method C of Example 8 (30.8 g) is lightly treated with 300 ml anhydrous ethanol in a 2 liter autoclave. 1.54 g (50% wettability) of 5% Pd / C is added and the mixture is hydrogenated at 4 times atmospheric pressure for 1 hour to complete the absorption of hydrogen within this time. The catalyst is recovered by filtration, first on filter paper and then on 0.45 micron nylon millicore, using 100-150 ml ethanol for delivery and washing. The synthesized filtrate and washings were stripped to give a wet white solid, which was dissolved in 150 ml of a mixture of anhydrous ethanol and acetonitrile in a ratio of 1: 10, purified by hot filtration, boiled to 35 ml, and then Cool slowly to room temperature, granulate and filter to give 20.1 g (94%) of crystalline, concentrated, non-electrostatic title product. This title product is characterized by having sharp peaks that are mostly well resolved by IR (Newsol Mullet) analysis at the following points: 3340, 3300, 2900, 2836, 1725, 1650, 1628, 1580, 1532, 1455, 1410, 1370, 1280, 1240, 1216 and 1175 cm ^-1 .

[Method C]

The crystalline product (9.4 g) prepared immediately according to Method B was heated to reflux for 1 hour and dissolved in 1000 ml acetone. Cool the solution to room temperature and spawn a small amount of Method B product to induce crystallization. After stirring for 6 hours, filtered with a small amount of acetone wash and vacuum dried at 35 ° C. to recover 7.25 g of a further purified title product having the same IR peak as the peak of acetonitrile / ethanol crystals of Method B.

[Method D]

The product of Example 10 (0.50 g) was synthesized with 0.026 g of 5% Pd / C (50% wettability) in 125 ml anhydrous ethanol in an all-par hydrogenation vessel. The mixture is hydrogenated under tetrahydrogen atmosphere for 2.5 hours. The catalyst is recovered by filtration and the filtrate is stripped to afford the title compound as a sticky solid which is crystallized according to the previous method.

Example 11

N- (R-3-methyl-4-heptenoyl) -D-gamma-glutamyl-glycyl-D-alanine.

The product of Example 9 (1 g) is dissolved in 5 ml CH ₃ OH. ¹ N NaOH (2.50 ml) is added and the mixture is stirred at circulating temperature for 3 hours. Strip CH ₃ OH and dilute the aqueous residue with 7.5 ml H ₂ O, extract twice with 7.5 ml ethyl acetate and acidify to pH 3.0 with 1 N HCl. The acidified aqueous solution is extracted successively with fresh ethyl acetate and the extract is stripped to yield the title product, which is converted to lyophylate according to Method A of Example 10.

Example 12

R-trans-4-heptene-3-ol

The title product (10 g, 0.088 moles) of Preparation Example 5 is converted to the title product by the method of Example 1, except that the reaction temperature is -20 ° C in all steps during the synthesis of the reagents. After all reagents have been synthesized, the mixture is stirred at -20 ° C for 1.5 hours, warmed to room temperature, quenched by dropwise addition of 25 ml H ₂ O and quenched with 6 ml 30% NaOH (saturated with NaCl). . The quenched mixture is stirred for 20 minutes, diluted with 30 ml CH ₂ Cl ₂ and the aqueous layer is separated and washed twice with 50 ml fresh CH ₂ Cl ₂ .

The organic layer is synthesized, dried over MgSO ₄ and distilled to a 50 ml residual volume, which residue is chromatographed using pentane over silica gel, followed by 4: 1 pentane: ether as eluent. Discard the initial fraction containing less polarized impurities. The pure product fraction, R _f 0.3 (4: 1 pentane: ether), is synthesized and stripped to yield the title product as a clear colorless liquid (5.7 g).

Example 13

R-3-ethyl-4-heptenoic acid

By the method A of Example 3 except for using ether instead of CH ₂ Cl _{2 for} separation, the product of Example 12 (5.7 g, 0.05 mole) was condensed with ethyl orthoacetate, rearranged and saponified This title product is obtained as 1.4 g of oil: TLC R _f 0.4 (2: 1 hexanes: ether), the intermediate edyl ester, shows R _f 0.25 in the same TLC system.

Example 14

S-3-ethyl heptanoic acid

(CH₃OH내에 C=2%).The product of Example 13 (1.4 g) is hydrogenated in 20 ml CH ₃ OH on 0.1 g 5% Pd (OH ₂ ) / C for 1 hour at 4 times atmospheric pressure. The catalyst is recovered by filtration. The filtrate is stripped in solvent and the residue is distilled off to yield 1.0 g of the title product: boiling point 76-77 ° C./0.4 torr:

(C = 2% in CH ₃ OH).

Example 15

S-3-ethylheptanoyl chloride

Under dry N ₂ , the product of Example 14 (1.0 g, 0.00633 mol) is dissolved in 10 ml CH ₂ Cl ₂ . Oxalyl chloride (0.547 ml, 0.00627 mol) is added and the mixture is stirred for 1 hour, when gas evolution ceases. The resulting solution of the title product is evaporated under a stream of N ₂ to 7 ml and used directly in the next step.

In the same way, the unsaturated acid of Example 13 is converted to a solution of R-3-ethyl-4-heptanoyl chloride.

Example 16

N- (S-3-ethylheptanoyl) -D-gamma-glutamyl (alphabenzyl ester) -glycyl-D-alanine benzyl ester Formulation of the title product of example 4 (1.1 g, 0.00222 mol) in 30 ml CH ₂ Cl ₂ and triethylamine (0.935 ml, 0.00666 mol). A CH ₂ Cl ₂ solution of S-3-ethylheptanoyl chloride (2.3 mol, 0.00211 mol) of Example 15 was added and the mixture was stirred for 0.75 h under N ₂ , followed by 10% HCl, H ₂ O, 10 Wash with 20 ml of% K ₂ CO ₃ and brine, dry over MgSO ₄ and strip to obtain a solid residue, triturate with ether and recover 0.8 g by filtration. The latter material was taken up in ethyl acetate and rewashed as above to obtain 0.7 g of a second solid under restripping, which was chromatographed on silica gel using CHCl ₃ : CH ₃ OH of 97: 3 as eluent. Completion yields 467 mg of the title product purified.

In the same way, 3-ethyl-4-heptenoyl chloride was coupled with the title product of Preparation Example N- (R-3-ethyl-4-heptenoyl) -D-gamma-glutamyl (alpha benzyl ester) -Glysyl-D-alanine benzyl ester is obtained.

Example 17

N- (S-3-ethylheptanoyl) -D-gamma-glutamyl-glycyl-D-alanine

The title product (467 mg) of the previous example is hydrogenated by the example 14 method. After recovering the catalyst, the filtrate is stripped with foam, dissolved in H ₂ O, filtered and lyophilized to give the title product (238 mg).

¹ H-nmr (DMSO-d ₆ ) delta (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 ).

In the same way the unsaturated product of the previous example is converted to the same product.

Example 18

N- (R-3-ethyl-4-heptenoyl) -D-gamma-glutamyl-glycyl-D-alanine

The unsaturated product of Example 16 is hydrolyzed by the method of Example 11 to afford the title product.

Example 19

N- (S-3-ethylheptanoyl) -D-gamma-glutamyl (alpha benzyl ester) -glycyl-D-alanine butyl ester

By the method of Example 17, the S-3-ethylheptanoyl chloride and D-gamma-glutamyl (alpha benzyl ester) -glycyl-D-alanine butyl ester (1.0 g, 0.00222 mol) of Example 16 were coupled Ring to produce the title product, isolate and purify a similar process. Yield amount is 0.7g.

¹ N-nmr (DMSO-d ₆ ) delta (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).

In the same manner, R-3-ethyl-4-heptanoyl chloride was replaced with N- (R-3-ethyl-4-heptanoyl) -D-gamma-glutamyl (alpha benzyl ester) -glycyl-D-alanine butyl ester Switch to

Example 20

N- (S-3-ethylheptanoyl) -D-gamma-glutamyl-glycyl-D-alanine butyl ester

By the method of Example 14, the title product of the previous example was hydrogenated to produce the title product. Following recovery of the catalyst, the filtrate is triturated with ether and filtered to strip off the recovered solid (256 mg, melting point 130-131 ° C.).

In the same way, the unsaturated product of the previous example is converted to the same product.

Example 21

R-3-ethyl-4-heptenal

Hg (OAc) ₂ was successively 5 times (2 g per 2 g, 2 h) while heating the solution of R-trans-4-hepten-3-ol (3.0 g) of Example 12 in reflux in 300 ml of dry ethyl vinyl ether. At intervals). After further heating to reflux for 18 hours, the clear solution is cooled to room temperature, treated with 0.5 ml of acetic acid on ice and stirred for 3 hours. The resulting solution is diluted with ether and poured into 200 ml 5% KOH solution and extracted three times with ether. The synthesized ether extract is then dried over K ₂ CO ₃ and stripped to give allyl vinyl ether, R-trans-O-vinyl-4-hepten-3-ol.

The solution of allyl vinyl ether (2.3 g) in 20 ml decalin is refluxed for 10.5 hours. The solution obtained is then cooled to room temperature and placed directly on a column of silica gel. Hexane flowed out to remove decalin, and then the title product was recovered by elution of the column with ether and stripping.

Example 22

S-3-ethyl-4-heptenoic acid

The product of the previous example (100 mg) is dissolved in 4 ml of acetone and cooled to 0-5 ° C. In a separate flask, CrO ₃ (72.1 g, 0.072 mol) with 50 ml H ₂ O was mixed and stirred at 0-5 ° C. and 62.1 ml of concentrated H ₂ SO ₄ were slowly added and the mixture diluted in 250 ml H ₂ O Obtain H ₂ CrO ₄ (Jones reagent) in 2.88M solution. The latter solution (1 ml) is slightly seeded into the acetone solution for 1 hour. When the temperature rises, the reagent is added and maintained at 17 to 25 ° C. The mixture was recooled to 6 ° C. and 70 ml of 2-propanol was added for 10 minutes (without raising the temperature to 20 ° C. during this process), then concentrated in vacuo to oil and stirred for 2.5 N for 50 minutes. 8 ml of NaOH is added and maintained at 22 ± 5 ° C. The mixture is heated and filtered over hot diatomaceous earth as 2.5 N NaOH washes. The synthesized filtrate and washings are extracted three times with 300 ml isopropyl ether. The synthesized organic layer is back extracted with 200ml 2N NaOH. The synthesized aqueous layer was acidified to pH 1.0 by slowly adding 0.5 ml of concentrated HCl and the product was extracted three times with 300 ml of fresh isopropyl ether. Organic extracts are synthesized and extracted to give the title product as an oil.

Example 23

S-7- (t-butyl dimethylsilyloxy) -5-methyl-1-heptene

), 4.8-5.3(m, =

), 3.7(t, J=6.5Hz,

), 0.08(s, C(

)₃) 및 0.0(s, Si(

)₂), 8mol%(C₆H₅)₃PO(7.6, s, 1.25H)로 오염된 물질.In a 500 ml, four-necked round bottom flask equipped with a stirrer, thermometer, N _{2 portion} and addition funnel, (methyl) triphenylphosphonium bromide (25.7 g, 0.072 mol, 1.25 equiv) was added in 77 ml THF (tetrahydrofuran) Treat lightly and cool in acetone / wet ice bath. Add n-butyllithium (43.2 ml of 1.6N, 0.069 mol, 1.20 equiv. in hexane) to the addition funnel. Initially treat lightly at -8 ° C and keep at ± 1 ° C with butyl lithium added for about 1 hour as the temperature rises. The aldehyde product of Preparation Example 2 (14.1 g, 0.0567 mol) in 14 ml THF was added at 40 ° C in small portions for 40 minutes. After stirring for an additional 15 minutes, it can be seen that the starting aldehyde is not detected by TLC with 3: 1 hexanes: ether as eluent (Rf starting aldehyde 0.6: Rf product 0.95). The reaction mixture is warmed to circulating temperature and diluted with 150 ml ethyl acetate and 90 ml H ₂ O. The organic layer is separated and washed twice with 100 ml H ₂ O. Three aqueous layers are synthesized and backwashed with 40 ml ethyl acetate. The organic layer was synthesized, dried over MgSO ₄ and stripped to give 25 g of oil, which was ground with 10 ml hexane and filtered on a sinter glass funnel, the solid was properly repulped four times with 10 ml hexane and the synthesized hexane Strip the extract and washes to afford the title product (13.5 g, 96.6%) as an oil: ¹ H-nmr (CDCl ₃ ) delta (ppm): 5.4-6.2 (m, =

), 4.8-5.3 (m, =

), 3.7 (t, J = 6.5 Hz,

), 0.08 (s, C (

) ₃ ) and 0.0 (s, Si (

) ₂ ), material contaminated with 8 mol% (C ₆ H ₅ ) ₃ PO (7.6, s, 1.25H).

Example 24

S-3-methylhept-6-en-1-ol

[Method A]

By the method of the previous example (except using 2.2 equivalents each of (methyl) triphenyl phosphonium bromide and n-butyllithium), the aldehyde product of Formulation Example 6 (26.3 g, 0.20 mol; Amount) is reacted with methylenetriphenylphosphoran. Reformation of the solid rubber is detected during the addition of the aldehyde solution, but once warmed to circulating temperature for operation, the reaction becomes a pale slurry. The reaction mixture is diluted with 500 ml H ₂ O and 300 ml ethyl acetate. The layers are separated and washed three times with 250 ml H ₂ O. The synthesized aqueous layer was backwashed with 300 ml ethyl acetate. Three organic layers were synthesized, dried over MgSO ₄ and stripped to yield 65.7 g of oil containing 25.6 g of the title product (100%) and about 40 g of triphenylphosphine oxide, which was found in Example 4 above. It is suitable for further process.

The pure title product can be more easily obtained with conventional hydrolysis of the product of Example 23, for example the dilute sulfuric acid of Example 26. The title product is separated into ethyl acetate by extraction, dried over MgSO ₄ and stripped.

Example 25

S-3-ethyl-6-heptene

The title product (1.14 g, 0.01 mol) of the previous example is dissolved in 20 ml CH ₂ Cl ₂ and cooled to 0 ° C. Pyridinium chlorochromate (4.30 g, 0.02 mol) is added to maintain the temperature at 0-5 ° C. The mixture is warmed to circulating temperature, stirred for 2 hours, filtered through a pad of silica gel and the filtrate stripped to give the title product as an oil, which is further purified by distillation as necessary.

Example 26

S-3-methyl-6-heptenoic acid

[Method A]

), 5.8(m, =

, 5.08m, =

) 및 1.0(d,

) 및 이소프로필 에테르 8.6몰%(6.3중량%)와 오염된 것을 보여주는 3.7 및 1.1에서 이소프로필 에테르 피크를 갖게되며, 상기 오염은 추가의 과정에서 본 생성물의 사용에 방해하지는 않는다.In a 2000 ml three-neck round bottom flask equipped with a stirrer, thermometer and addition funnel, the title product of Example 23 (81 g, exact amount for pure components, 0.33 mol) is dissolved in 400 ml acetone and cooled to 0-5 ° C. In a separate flask, CrO ₃ (72.1 g, 0.72 mol) was mixed with 50 ml H ₂ O, stirred at 0-5 ° C., 62.1 ml concentrated H ₂ SO ₄ was added slowly and the mixture diluted to 250 ml as H ₂ O to 2.88 MH. ₂ CrO ₄ , Jones reagent). The latter solution (240 ml, 0.67 mol) is added in small portions to the acetone solution for 1.2 hours. If the temperature rises sharply to 17 ° C., it is maintained at 17 to 25 ° C. while adding a reagent. If fever does not occur, stop the addition. To the mixture was added 70 ml 2-propanol for 10 minutes (while the temperature was raised to 20 ° C.) and recooled to 6 ° C., then concentrated to oil as a vacuum, and 400 ml 5N NaOH was added to the oil for 50 minutes with stirring. Maintain at ± 5 ° C. The cloudy reaction mixture is diluted with 400 ml of H ₂ O and filtered over diatomaceous earth. The wet cake is repulped in 400 ml H ₂ O and 50 ml 5N NaOH and warmed again in a steam bath. The synthesized filtrate is washed three times with 300 ml isopropyl ether. The synthesized organic layer is back extracted as 200ml 2N NaOH. The synthesized aqueous layer is acidified to pH 1.0 by slow addition of 50 ml concentrated HCl and the product is extracted three times with 300 ml fresh isopropyl ether. The organic extract is synthesized and stripped to yield 29.1 g (61%) of the title product as an oil. A second separation of the filter cake on diatomaceous earth followed by similar separation yields 7.7 g (16%) of additional product. The ¹ H-nmr of the synthesized product (CDCl ₃ ) delta (ppm) is 11.9 (s,

), 5.8 (m, =

, 5.08 m, =

) And 1.0 (d,

) And isopropyl ether with isopropyl ether peaks at 3.7 and 1.1 showing contamination with 8.6 mole% (6.3 wt.%), The contamination does not interfere with the use of the product in further processes.

[Method B]

-3-메틸헵트-6-엔-1-올의 26.3g, 0.20몰을 함유하는 65.7g)을 본 실시예의 방법 A에 따라 산화시킨다. 이소프로판을 급냉하여 1시간 동안 0 내지 5℃에서 교반하면, 이 시간내에 반응 혼합물이 완전히 녹색이 되며, 유기 용매를 스트립핑하고, 수성 잔사를 250ml H₂O로 희석시키고 이소프로필 에테르 250ml로 3회 추출한다. 유기 추출물을 합성하고 160ml 2N NaOH로 처리하면, 1N NaOH 세척액을 통해 여과에 의해 회수되는 (C₆H₅)₃PO의 중침전(출발물질에서 오염 성분)이 일어난다. 여과액 및 세척액을 합성하고, 층을 분리하고 유기층을 80ml 추가 1N NaOH로 세척시킨다. 모든 수성층을 합성하고, 250ml 이소프로필 에테르로 3회 세척하고 50ml 진한 HCl로 pH 1.0이 되도록 산성화시키고 원하는 생성물을 250ml 신선한 이소프로필 에테르로 3회 추출한다. 합성된 산 유기 추출물을 MgSO₄상에서 건조시키고 스트립핑시켜 표제 생성물 14.0g을 수득하며, 필요에 따라 증류에 의해 추가로 정제한다.Product of Method A of Example 24

26.3 g of -3-methylhept-6-en-1-ol, 65.7 g containing 0.20 mol) are oxidized according to Method A of this example. When the isopropane was quenched and stirred at 0-5 ° C. for 1 hour, the reaction mixture became completely green within this time, the organic solvent was stripped off, the aqueous residue diluted with 250 ml H ₂ O and 3 with 250 ml of isopropyl ether. Extract times. Synthesis of organic extracts and treatment with 160 ml 2N NaOH result in heavy precipitation (contaminant in the starting material) of (C ₆ H ₅ ) ₃ PO recovered by filtration through 1N NaOH wash. The filtrate and washes are synthesized, the layers are separated and the organic layer is washed with 80 ml additional 1N NaOH. All aqueous layers are synthesized, washed three times with 250 ml isopropyl ether, acidified to pH 1.0 with 50 ml concentrated HCl and the desired product is extracted three times with 250 ml fresh isopropyl ether. The synthesized acid organic extract is dried over MgSO ₄ and stripped to give 14.0 g of the title product, which is further purified by distillation as necessary.

Example 27

S-3-methylheptanoic acid

10% Pd / C (1.64 g, 50% wettability), 150 ml ethyl acetate and unsaturated acid (3.28 g, exact amount for pure components) were added to a par hydrogenation vessel and the slurry was added at 4 atmospheres. Hydrogenation for 1.5 hours completes H ₂ uptake within this time. The catalyst is recovered by filtration on diatomaceous earth and the filtrate is stripped to give 3.20 g (96%) of the title product as an oil.

Alternatively, in 5% Pd / C (2 g, 50% wettability) and 150 ml ethyl acetate, the title product of the previous example (33.3 g, exact amount for pure components) was placed in a 1 liter autoclave at 4 atmospheres, 30-31. Hydrogenation at 2 ° C. for 2 hours completes H ₂ uptake by this time. The catalyst is recovered by filtration on diatomaceous earth and the filtrate is stripped to give 35.4 g of oil which is distilled under high vacuum to give 29.6 g (87.6%) of the purified title product:

), 1.0(d,

), 0.6-2.8(m,잔기 13H) : ir(필름) 3400-2400, 2960, 2925, 2860, 1708, 1458, 1410, 1380, 1295, 1228, 1190, 1152, 1100, 930cm^-1

(CH₃OH내에서 C=1%) :

Boiling point ^{77-79 ℃ / 0.2mm: 1 H-} nmr (CDCl 3) (ppm): 12.0 (s,

), 1.0 (d,

), 0.6-2.8 (m, residue 13H): ir (film) 3400-2400, 2960, 2925, 2860, 1708, 1458, 1410, 1380, 1295, 1228, 1190, 1152, 1100, 930cm ^-1

(C = 1% in CH ₃ OH):

Example 28

S-3-ethyl-6-heptenoyl chloride.

The acid product of Example 4 (0.747 g, 5 mmol) was converted directly to a CH ₂ Cl ₂ solution of the title product by the method of Example 6 and used directly in Example 9. Alternatively, the reaction mixture is stripped to yield the title product, if necessary distilled under reduced pressure.

Example 29

-3-메틸-6-헵테노일)-D-감마-글루타밀-(알파 벤질 에스테르)-글리실-D-알라닌 벤질-에스테르N- (

3-Methyl-6-heptenoyl) -D-gamma-glutamyl- (alpha benzyl ester) -glycyl-D-alanine benzyl-ester

By method C of example 8, the product of preparation 4 (2.77 g, 5 mmol) is coupled with the total amount of acid chloride in CH ₂ Cl ₂ obtained from example 28. 2.77 g of the initially obtained product, which was recovered by stripping the washed and dried organic layer, was dissolved in 200 ml hot ethyl acetate, the solution was diluted with 20 ml hexane and purified by filtration, 2.24 g (77%, melting point: 137.5-) 139.5 ° C.).

Example 30

-3-메틸-6-헵테노일)-D-감마-글루타밀-글리실-D-알라닐-N- (

-3-methyl-6-heptenoyl) -D-gamma-glutamyl-glycyl-D-alanyl-

Example 2 g of product (1 g) is dissolved in 5 ml CH ₃ OH. ¹ N NaOH (2.50 ml) is added and the mixture is stirred at circulating temperature for 3 hours. Strip CH ₃ OH and dilute the aqueous residue with 7.5 ml H ₂ O and extract twice with 7.5 ml ethyl acetate and acidify with 1N HCl to pH 3.0. The acidified aqueous solution is successively extracted with fresh ethyl acetate and the extract is stripped to yield the title product which is converted to lyophilate according to method A of Example 10.

Example 31

-3-에틸-6-헵타노일)-D-감마-글루타밀(알파 벤질 에스테르)-글리실-D-알라닌 부틸 에스테르N- (

3-Ethyl-6-heptanoyl) -D-gamma-glutamyl (alpha benzyl ester) -glycyl-D-alanine butyl ester

D-gamma glutamyl (alpha benzyl ester) -glycyl-D-alanine butyl ester (Preparation Example 4) was prepared by the method A of Example 8 with S-3-methyl-6-heptenoyl chloride (Example 28). Coupling yields the title product.

Example 32

-3-메틸헵타노일)-D-감마-글루타밀-글리실-D-알라닌 부틸 에스테르N- (

3-Methylheptanoyl) -D-gamma-glutamyl-glycyl-D-alanine butyl ester

The hydrogenation reaction of Example 10 converts the product of the previous example to the title product.

[Production Example 1]

Racemic trans-4-hexene-3-ol

400 ml THF is placed in a 3-liter four-necked flask equipped with a stirrer, addition funnel, thermometer and nitrogen inlet. The THF is cooled to -70 ° C and ethyl magnesium bromide (600 ml of 2M in THF, 1.2 mol) is added. Crotonaldehyde (78 ml, 0.94 mole) is added over 7 minutes while maintaining the temperature between -70 ° C and -60 ° C. After 20 minutes of stirring at −70 ° C., if the mixture gradually warms to −20 ° C. for 1 hour, the mixture is recooled to −70 ° C. and carefully quenched with saturated NH ₄ Cl (initial foaming) and circulated. Warm to temperature and dilute with 200 ml CH ₃ COOH and 100 ml H ₂ O. The layer system is saturated with NaCl ₂ . The aqueous layer is separated and extracted twice with 500 ml ether. The original organic layer and ether extracts are synthesized, washed four times with 250 ml saturated NaHCO ₃ , dried over MgSO ₄ and stripped to distill the residue at atmospheric pressure:

Boiling point: 133-137 ° C., ¹ H-nmr (CDCl ₃ ) delta (ppm): 5.53 (complex m, 2H), 3.93 (m, 1H), 2.26 (s, 1H), 1.71 (d, 3H), 1.43 (m, 2 H), 0.84 (t, 3 H).

[Production Example 2]

Glysyl-D-alanine benzyl ester hydrochloride

-부틸옥시-카르보닐 글리신, 20g(57밀리몰)의 D-알라닌 벤질 에스테르 p-톨루엔 황산염 및 5.77g(57밀리몰)의 트리에틸아민을 함유하는 100ml 메틸렌 클로라이드의 냉각(℃) 용액에 12.3g(60밀리몰)의 디사이클로헥실카르보디이미드를 가하고 얻어진 반응 혼합물을 상온까지 상승하도록 따뜻하게 한다. 18시간후 혼합물을 여과하고 여과액을 진공상태로 농축시킨다. 잔사를 200ml의 에틸 아세테이트로 용해시키고 유기층을 2.5% 염산, 물, 포화 중탄산 나트륨 용액 및 소금 용액으로 세척한다. 유기층을 분리하고 황산 마그네슘상에서 건조시키고 감압하에서 증발시킨다. 얻어진 오일에 수소 염화물과 포화된 200ml의 디옥산을 가한다. 30분후 400ml의 디에틸 에테르를 가하고 생성물(10.9g(70% 수율))을 질소하에서 여과시킨다.10 g (57 mmol) of N-

12.3 g (100 ° C.) in a cold (° C.) solution of 100 ml methylene chloride containing butyloxy-carbonyl glycine, 20 g (57 mmol) of D-alanine benzyl ester p-toluene sulfate and 5.77 g (57 mmol) of triethylamine. 60 mmol) of dicyclohexylcarbodiimide is added and the reaction mixture obtained is warmed to rise to room temperature. After 18 hours the mixture is filtered and the filtrate is concentrated in vacuo. The residue is dissolved with 200 ml of ethyl acetate and the organic layer is washed with 2.5% hydrochloric acid, water, saturated sodium bicarbonate solution and salt solution. The organic layer is separated, dried over magnesium sulfate and evaporated under reduced pressure. Hydrogen chloride and saturated 200 ml dioxane are added to the obtained oil. After 30 minutes 400 ml of diethyl ether are added and the product (10.9 g (70% yield)) is filtered under nitrogen.

[Manufacture example 3]

-부톡시카르보닐-D-감마-글루타밀(알파벤질에스테르)하이드로석신아미드 에스테르N-

Butoxycarbonyl-D-gamma-glutamyl (alphabenzyl ester) hydrosuccinamide ester

-부톡시카르보닐-D-감마-글루탐산 알파-벤질 에스테르 및 17.3g(150밀리몰)의 N-하이드록시석신아미드를 함유하는 1500ml 메틸렌 클로라이드에 30.9g(15밀리몰)의 디사이클로헥실카르보디이미드를 가하고 얻어진 반응 혼합물을 18시간 동안 상온에서 교반시킨다. 고체를 여과하고 여과액을 진공상태에서 농축시킨다. 잔사를 디에틸 에테르와 동결건조시키고 고체(48.7g(68% 수율))을 질소하에서 여과한다.50 g (143 mmol) of N-

30.9 g (15 mmol) of dicyclohexylcarbodiimide in 1500 ml methylene chloride containing butoxycarbonyl-D-gamma-glutamic acid alpha-benzyl ester and 17.3 g (150 mmol) of N-hydroxysuccinamide The reaction mixture obtained is added and stirred at room temperature for 18 hours. The solid is filtered off and the filtrate is concentrated in vacuo. The residue is lyophilized with diethyl ether and the solid (48.7 g (68% yield)) is filtered under nitrogen.

[Production Example 4]

D-gamma-glutamyl (alpha benzyl ester) -glycyl-D-alanine benzyl ester hydrochloride

-부톡시카르보닐-D-감마-글루타밀(알파벤질 에스테르)하이드록시석신아미드 에스테르, 2.71g(9.92밀리몰)의 글리실-D-알라닌 벤질 에스테르 하이드로클로라이도 및 1.0g(9.92밀리몰)의 트리에틸아민을 함유하는 용액을 상온에서 18시간 동안 교반시킨후, 진공상태에서 농축시킨다. 잔사를 200ml의 에틸 아세테이트에 용해시키고 용액을 2.5% 염산, 물, 10% 탄산칼륨 및 소금물 용액으로 세척한다. 유기상을 분리하고, 황산 마그네슘상에서 건조시키고 감압하에서 증발시킨다. 잔사를 염화수소화 포화된 200ml의 디옥산으로 처리하고 2시간 동안 교반시킨다. 용액을 진공상태에서 건조상태가 되도록 농축시키고 잔사를 디에틸 에테르로 연마한다. 고체를 질소 3.41g(73%수율)하에 여과한다.4.3 g (9.45 mmol) N- in 100 ml methylene chloride

-Butoxycarbonyl-D-gamma-glutamyl (alphabenzyl ester) hydroxysuccinamide ester, 2.71 g (9.92 mmol) glycyl-D-alanine benzyl ester hydrochlorido and 1.0 g (9.92 mmol) tree The solution containing ethylamine is stirred at room temperature for 18 hours and then concentrated in vacuo. The residue is dissolved in 200 ml of ethyl acetate and the solution is washed with 2.5% hydrochloric acid, water, 10% potassium carbonate and brine solution. The organic phase is separated, dried over magnesium sulfate and evaporated under reduced pressure. The residue is treated with 200 ml of saturated hydrochloric acid dioxane and stirred for 2 hours. The solution is concentrated to dryness in vacuo and the residue is triturated with diethyl ether. The solid is filtered under 3.41 g (73% yield) of nitrogen.

In the same manner the product of the previous preparation and the glycyl-D-alanine butyl ester are converted to D-gamma-glutamyl (alpha benzyl ester) -glysil-D-alanine butyl ester.

Production Example 5

Racemic trans-4-heptene-3-ol

In the method of Preparation Example 1, 2-pentenal (25 g, 0.30 mole) is converted to the distilled title product (25.6 g, boiling point 145-150 ° C.) except using a conventional ether as solvent.

[Manufacture example 6]

0- (t-butyldimethylsilyl) -S-citronellol

In a 500 ml 4-neck round bottom flask equipped with a stirrer, thermometer, N ₂ inlet and addition funnel, S-citronellol (547 g, 3.5 mol) was dissolved in 547 ml DMF (dimethylformamide) at circulation temperature (21 ° C.). Let's do it. Imidazole (262, 1 g, 3.85 moles, 1.1 equiv) is added. The temperature is lowered to 13 ° C. and then further reduced to −6.5 ° C. with an ice / acetone bath. T-Butyl dimethylchlorosilane (580.3 g, 3.85 mol) was added to 1160 ml DMF over 1.25 hours, and then stirred at high speed to dissolve. Then, the temperature was gradually raised to 11 ° C. for the same time. After an additional 0.25 hour, tlc (3: 1 hexanes: ether) showed complete conversion to the desired product (Rf 0.2 of starting material: Rf 0.9 of product).

To separate, the mixture is added to 500 ml hexane and 1000 ml ice water. The layers are separated and the organic layer is washed with 2000 ml 0.25N NaOH cooled with ice. Two aqueous layers were synthesized and refluxed with 500 ml hexane and combined with the original organic layer, washed with 500 ml saturated NaHCO ₃ , dried over MgSO ₄ and stripped to give quantitative yield of the title product (986.7 g, trace retention of solvent). 104%)

), 3.65(t, J=6.5Hz, -O-

-)), 1.7과 1.65(2S, 2=C(

)), 0.8(s, -SiC(

)₃), 및 0.0(s,-Si(

)₂). ¹ H-nmr (CDCl ₃ ) delta (ppm): 5.2 (t, J = 7Hz =

), 3.65 (t, J = 6.5 Hz, -O-

) And 1.7 and 1.65 (2S, 2 = C (

)), 0.8 (s, -SiC (

) ₃ ), and 0.0 (s, -Si (

) ₂ ).

[Manufacture example 7]

S-6- (t-butyl dimethylsilyloxy) -4-methylhexanal

500 ml, four-necked round bottom flask equipped with mechanical stirrer, straight glass inlet for O ₃ / O ₂ stream, thermometer and outlet connected to saturated KI trap (81.2 g, exact figures for solvent content, 0.30 mole) is dissolved in 120 ml CH ₂ Cl ₂ and 81 ml CH ₃ OH. NaHCO ₃ (6.3 g, 0.075 mol, 0.25 equiv) is added and the mixture is cooled to -10 ° C in an acetone / dry ice bath. The temperature was further lowered to maintain -72 ° C to -75 ° C to bubble O ₃ / O ₂ into the reaction for 6 hours. In less than an hour, O ₃ is no longer dissolved in the reaction mixture, which can be seen by the yellow formation in the KI trap. As hexane eluent tlc shows the reaction is complete (Rf 0.3 of starting material: Rf 0.0 of intermediate).

The reaction mixture is purged with N ₂ with excess O ₃ , dimethyl sulfide (26.4 ml, 22.4 g, 0.36 mol, 1.2 equiv) is added, the water bath is removed and the mixture is warmed to circulating temperature and under N ₂ Stir for 16 hours, within which time tlc (6: 1 hexanes: ether) shows no intermediate remaining (Rf 0.8 of intermediate: Rf 0.05 of product). The mixture is then stripped and the residue is partitioned between 150 ml ethyl acetate and 300 ml H ₂ O. The organic layer is washed with 300 ml fresh H ₂ O. The synthesized H ₂ O layer is backwashed with 150 ml fresh ethyl acetate. The organic layer is synthesized, dried over MgSO ₄ and ultimately stripped for 16 hours under high vacuum to yield quantitative yield of the product as an oil (74.7 g, 101.9% of theory due to small amount of solvent contamination).

), 3.6(t,J=6Hz, -O-

-), 0.9(s, -SiC(

)₃), 0.0(s, -Si(

)₂) . ¹ H-nmr (CDC ₁₃ ) delta (ppm): 9.75 (t,

), 3.6 (t, J = 6Hz, -O-

-), 0.9 (s, -SiC (

) ₃ ), 0.0 (s, -Si (

₂ ).

[Manufacture example 8]

S-6-hydroxy-4-ethylhexanal

[Method A]

A gas inlet and outlet tube connected to a magnetic stirrer, thermometer, gas-washing vessel containing saturated KI is placed in a 250 ml three-neck round bottom flask. S-citronellol (31.25 g, 0.20 mole) was added to the flask in 81 ml CH ₂ C ₁₂ and 54 ml CH ₃ OH and cooled to -8 ° C. Bubble O ₂ / O ₃ through the reaction for 4.5 hours while maintaining the temperature between -2 ° and -10 ° C, showing trapping of excess O ₃ by KI solution within this time The reaction was completed by the Positive Starch / KI Paper test.

The reaction mixture is held at −5 ° C., purged with N ₂ and methyl sulfide (17.7 ml, 15.0 g, 0.24 mol) is added. The reaction mixture was warmed to circulating temperature and stirred for 16 hours, indicating that tlc using isopropyl ether as eluent within this time showed the reaction was complete (Rf 0.7 of s-citronellol: Rf 0.4 of product). Finally, the solvent is stripped to give 50.8 g of oil. The oil is diluted with 30 ml of ethyl acetate and 25 ml H ₂ O and a single phase is obtained. Additional 150 ml ether is added to give a biphasic phase. The layers are separated and the inorganic phase is washed twice with 25 ml H ₂ O, dried over MgSO ₄ and stripped to give 23.9 g of a colorless oil. The synthesized aqueous layer is extracted twice with 50 ml ethyl acetate and the extracts are synthesized, dried and stripped to yield 7.8 g of additional colorless oil. The colorless oil is synthesized and further stripped to yield the title product (28.3 g, 108.7% of theory), with tlc (specified above) suitable for use in further reactions as described above except solvent contamination. It can be clearly seen.

[Method B]

The product of Preparation Example 2 is hydrolyzed by conventional methods such as dilute sulfuric acid of Example 4. The title product is separated by extraction with ethyl acetate and simultaneously stripped as in method A above.

Claims (21)

(a) oxidizing S-enantiomers in the presence of titanium tetraisopropoxide and L-(+)-diisopropyl tartrate and unreacted trans-R-4-hexen-3-ol or trans- Reacting racemic trans-4-hexene-3-ol or trans-4-hepten-3-ol with tert-butyl hydroperoxide in an amount sufficient to maintain R-4-hepten-3-ol; (b) to the trans -R-4- cyclohexene-3-ol or trans -R-4- heptene-3-ol tree [(C ₁ -C ₃₎ alkyl] orthoacetate in the presence of an acid and a condensation (C _1- C ₃ ) alkyl R-3-methyl-4-heptenoate or R-3-ethyl-4-heptenoate ester; (c) hydrolyzing the (C ₁ -C ₃ ) alkyl ester to form the R-3-methyl-4-heptenoic acid or R-3-ethyl-4-hepteno acid Method for preparing methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid. (a) oxidizing S-enantiomers in the presence of titanium tetraisopropoxide and L-(+)-diisopropyl tartrate and unreacted trans-R-4-hexen-3-ol or trans- Reacting racemic trans-4-hexene-3-ol or trans-4-hepten-3-ol with tert-butyl hydroperoxide in an amount sufficient to maintain R-4-hepten-3-ol; (b) to the trans -R-4- cyclohexene-3-ol or trans -R-4- heptene-3-ol tree [(C ₁ -C ₃₎ alkyl] orthoacetate in the presence of an acid and a condensation (C _1- C ₃ ) alkyl R-3-methyl-4-heptenoate or R-3-ethyl-4-heptenoate; (c) ester hydrolysis of the (C ₁ -C ₃ ) alkyl ester to form R-3-methyl-4-heptenoic acid or R-ethyl-4-heptenoic acid; (d) catalytic hydrogenation of the 4-heptonoic acid to produce the S-3-methylheptanoic acid or S-3-ethylheptanoic acid; (e) catalytic hydrogenation of the (C ₁ -C ₃ ) alkyl ester to produce (C ₁ -C ₃ ) alkyl S-3-methylheptanoate or S-3-ethylbutanoate; (f) ester hydrolyzing the S-3-methylheptanoate or S-3-ethylheptanoate ester to produce the S-3-methylheptanoic acid or S-3-ethylheptanoic acid Method for preparing S-3-methyl-heptanoic acid. The process according to claim 1 or 2, wherein the (C ₁ -C ₃ ) alkyl groups are each ethyl and the acid is AlCl ₃ . (a) oxidizing S-enantiomers in the presence of titanium tetraisopropoxide and L-(+)-diisopropyl tartrate and unreacted trans-R-4-hexen-3-ol or trans- Reacting racemic trans-4-hexene-3-ol or trans-4-hepten-3-ol with tert-butyl hydroperoxide in an amount sufficient to maintain R-4-hepten-3-ol; (b) reacting the trans-R-4-hexene-3-ol or trans-R-4-hepten-3-ol with ethyl vinyl ether in the presence of Hg (CH ₃ CO ₂ ) ₂ to obtain the corresponding O-vinyl. To generate an ether; (c) rearranging the O-vinyl ether by heating in a reaction inert solvent to produce R-3-methyl-4-heptenal or R-3-ethyl-4-heptenal; (d) oxidizing the 4-heptenal with chromium anhydride in dilute inorganic acid to form the R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid; Process for the preparation of methyl-heptenoic acid or R-3-ethyl-4-heptenoic acid. The method of claim 4 wherein the inorganic acid is sulfuric acid. (a) S-6-hydroxy-4-methylhexanal or S-6- (silylprotected) hydroxy-4-methylhexanal is reacted with methylene triphenylphosphorane in a reaction inert solvent to give S-7- Forming hydroxy-5-methyl-1-heptane or S-7- (silylprotected) hydroxy-5-methyl-1-heptene; (b) if the product of step (a) contains a silyl protecting group, the hydrolysis of the protecting group in a dilute inorganic acid is carried out separately or concurrently with the subsequent step to give said S-7-hydroxy-5-methyl To form 1-heptene; (c) oxidizing the S-7-hydroxy-5-methyl-1-heptene with chromium anhydride in a dilute inorganic acid to form S-3-methyl-6-heptenoic acid. Method for preparing 6-heptenoic acid. 7. The process of claim 6 wherein S-6-hydroxy-4-methylhexanal is the reactant in step (a) and the inorganic acid is sulfuric acid. 7. The compound of claim 6, wherein S-6- (silylbro) hydroxy-4-hexanal is the reactant in step (a), steps (b) and (c) are performed simultaneously, and the silylprotecting group is tertiary- Butyl dimethylsilyl and the inorganic acid is sulfuric acid. (a) S-6-hydroxy-4-methylhexanal or S-6- (silylprotected) hydroxy-4-methylhexanal is reacted with methylene triphenylphosphorane in a reaction inert solvent to give S-7- Hydroxy-5-methyl-1-heptene or S-7- (silylprotected) hydroxy-5-methyl-1-heptene is formed; (b) if the product of step (a) contains a silyl bromo group, the hydrolysis of the protecting group in an inorganic acid is carried out separately or concurrently with the subsequent step of the S-7-hydroxy-5-methyl-1 -Heptene is formed; (c) oxidizing the S-7-hydroxy-5-methyl-1-heptene with chromium anhydride in a dilute inorganic acid to form S-3-methyl-6-heptenoic acid; (d) forming the S-3-methylheptenoic acid by catalytic hydrogenation of a double bond in the 6-heptenoic acid. 10. The process of claim 9 wherein S-6-hydroxy-4-methylhexanal is the reactant in step (a) and the inorganic acid is sulfuric acid. 10. The process of claim 9, wherein S-6- (silylprotected) -hydroxy-4-hexanal is the reactant in step (a), steps (b) and (c) are carried out simultaneously, and the silyl protecting group is tertiary. -Butyldimethylsilyl and the inorganic acid is sulfuric acid. (a) The active form of R-trans-3-methyl-4-heptenoic acid, R-trans-3-ethyl-4-hepteno or S-3-methyl-6-heptenoic acid is converted to an absolute stereochemical formula (II). Coupling with a compound of to form a compound of absolute stereochemical formula (IIIc); (b) A process for preparing a compound of formula (I), wherein the compound of formula (IIIc) is hydrolyzed to reduce double bonds and at the same time to hydrolyze benzyl ester group (s).

Wherein R is methyl or ethyl and R ⁴ and R ⁵ are each hydrogen or one of R ⁴ and R ⁵ is hydrogen and the other is (C ₁ -C ₆ ) alkyl or (C ₆ -C ₈ ) Cycloalkylmethyl, R ⁶ and R ⁷ are benzyl, or one of R ⁶ and R ⁷ is benzyl and the other is (C ₁ -C ₆ ) alkyl or (C ₆ -C ₈ ) cycloalkylmethyl, One or the other (but not both) of the dashed line represents a double bond and if the double bond is at the terminal 6,7-position, R is methyl. 13. The process of claim 12 wherein the active form of the acid is acid chloride, R ⁶ and R ⁷ are both benzyl and R ⁴ and R ⁵ are both hydrogen. A compound of absolute stereochemical formula (IV) or a pharmaceutically acceptable cation salt thereof wherein R ² and R ³ are hydrogen.

Wherein one or the other of the dashed lines represents a double bond and R is methyl or ethyl; R ² and R ³ are each hydrogen or are each independently (C ₁ -C ₆ ) alkyl, (C ₆ -C ₈ ) cycloalkylmethyl or benzyl, provided that the double bond is at the terminal groups 6, 7- If present, R is methyl. 15. The compound of claim 14, wherein R ² and R ³ are each hydrogen. Crystalline compound of absolute stereochemical formula (V).

R-trans-4-hexene-3-ol or R-trans-4-hepten-3-ol. Compounds of absolute stereochemical formula (VI) or compounds of other active forms of acids wherein X is OH.

Wherein R is methyl or ethyl, X is hydrogen, OR ¹ , OH or C ₁ and R ¹ is (C ₁ -C ₃ ) alkyl. The compound of claim 18, wherein X is OH or C ₁ . A compound of the absolute stereochemical formula or a compound of another active form of an acid wherein R ⁹ is -COOH.

Wherein R ⁹ is —CH ₂ OR ¹⁰ , —CH ₂ OH, —CHO, —COOH or —COC ₁ , and R ¹⁰ is hydrogen or a conventional silyl hydroxy protecting group. The compound of claim 20, wherein R ⁹ is —COOH or —COC ₁ .