KR20110073473A - Peptide compound and method for producing same - Google Patents

Abstract

The present invention provides an efficient production method that is excellent in terms of operation surface, purity and the like for industrial production.
Further, the present inventors have diligently studied about a highly efficient production method capable of obtaining an intermediate of high operability and high purity aimed at an industrial production method. As a result, the number of steps of leaving an amino acid can be shortened. To find a method for obtaining intermediates with high purity that are not purified by chromatography and to find a method for producing a cyclic peptide compound known to have anti-HCV activity, a peptide intermediate useful for the production thereof, and a method for producing the intermediate. The present invention was completed.
That is, in the present invention, since Edman decomposition is not repeated, the peptide intermediate and the cyclic peptide compound can be supplied in large quantities without performing the chromatography step according to the reaction.

Description

Peptide compound and its manufacturing method {PEPTIDE COMPOUND AND METHOD FOR PRODUCING SAME}

The present invention provides a method for preparing a cyclic peptide compound useful as an active ingredient of a pharmaceutical composition, in particular, a pharmaceutical composition for treating hepatitis C virus (HCV) known as an RNA replication inhibitor of hepatitis C virus replicon, an intermediate useful for the preparation thereof, and an intermediate thereof. It relates to a manufacturing method.

It is reported that the cyclic peptide compound of the following formula (A) or a salt thereof is useful as an active ingredient of a pharmaceutical composition for treating HCV based on the activity of inhibiting RNA replication of hepatitis C virus replicon (Patent Document 1). .

[Refer to Patent Document 1 for symbols in the formula]

Patent Document 1 discloses that a compound of Formula (A) or a salt thereof can be produced via a compound of Formula (I) or a salt thereof using starting material FR901459 (Patent Document 2) as a starting material. Specifically, it passed through the process of the following <1>-<6>.

<1> Preparation of a compound of formula (I) or a salt thereof by ring-opening after N-O translocation of a material or a salt thereof.

<2> Preparation of a compound of formula (A2-2) or a salt thereof by condensing threonine at the C terminus of a compound of formula (I) or a salt thereof.

<3> Preparation of the compound of the formula (A3) or a salt thereof by Edman decomposition three times with respect to the compound of the formula (A2-2) or a salt thereof.

<4> Preparation of a compound of formula (A4-4) or a salt thereof by condensing a compound of formula (A3) or a salt thereof with a diamino acid at the N-terminus.

<5> Preparation of a compound of formula (A) or a salt thereof by cyclization of a compound of formula (A4-4) or a salt thereof.

<6> Preparation of a compound of formula (Ab) or a salt thereof by reducing a compound of formula (Aa) or a salt thereof.

Each process is explained in full detail below.

<1> It is the process of manufacturing the compound of general formula (I) or its salt from FR901459 substance or salt thereof represented by general formula (A1-1). Specifically, a compound of formula (I) or a salt thereof is prepared by hydrolyzing a compound of formula (A1-2) or a salt thereof obtained by the protection of an amino group after a NO potential reaction to a FR901459 material under weakly acidic conditions. It was.

[Wherein, Prot ⁰ means a protecting group, and in the formula, other symbols refer to Patent Document 1]

<2> A process for producing the compound of formula (A2-2) or a salt thereof from the compound of formula (I) or a salt thereof. A compound of formula (A2-2) or a salt thereof was prepared by condensing a compound of formula (A2-1) with a compound of formula (I) or a salt thereof.

[In formula, Prot ⁰ means a protecting group and the other symbols refer to patent document 1]

<3> A process for producing the compound of formula (A3) or a salt thereof from the compound of formula (A2-2) or a salt thereof. The compound of formula (A3) or its salt is subjected to three times Edman decomposition to the compound of formula (A2-2) or salt thereof, that is, by sequentially leaving amino acids from the N terminus of the compound of formula (A2-2) or salt thereof Salts were prepared.

[Refer to Patent Document 1 for symbols in the formula]

<4> A process for preparing the compound of formula (A4-4) or a salt thereof from the compound of formula (A3) or a salt thereof. Condensed protected amino acid of formula (A4-3) with a compound of formula (A4-2) or a salt thereof obtained by carrying out the introduction and deprotection of the amino acid to the N terminus of the compound of formula (A3) or a salt thereof, The compound of formula (A4-4) or a salt thereof was prepared.

[Wherein, Prot ³ and Prot ⁴ are both protecting groups and other symbols refer to Patent Document 1]

<5> A process for preparing the compound of formula (Aa) or a salt thereof from the compound of formula (A4-4) or a salt thereof. After deprotecting the compound of formula (A4-4) or a salt thereof, the compound of formula (Aa) or a salt thereof was prepared by macrolactamization.

[Refer to Patent Document 1 for symbols in the formula]

<6> A process for producing the compound of formula (Ab) or a salt thereof from the compound of formula (Aa) or a salt thereof. By conventional catalytic reduction, the compound of formula (Ab) or a salt thereof is prepared.

[Refer to Patent Document 1 for symbols in the formula]

In the above-described manufacturing method, a problem when scaling up the compound of formula (A) or a salt thereof toward the industrial scale is a step of repeatedly performing Edman decomposition. Purification is necessary because the by-products obtained by Edman decomposition may complicate the next reaction. In particular, it is preferable not to perform column chromatography that requires complicated operations when expanding the scale toward an industrial scale. In this patent document 1 (for example, Prep 3), it went through the process refine | purified by column chromatography.

International Publication WO2007 / 049803 Japanese Patent Laid-Open No. 5-271267

Provided are a process for producing a compound of the general formula (A) having good efficiency in terms of operability and purity for industrial production, an intermediate useful for the preparation thereof, and a method for producing the same.

MEANS TO SOLVE THE PROBLEM The present inventors earnestly examined about the manufacturing method which is excellent in terms of the operability and purity which are aimed at the industrial manufacturing method, and finds the reaction which can shorten a process, and does not refine | purify the process water, even if it finds a reaction which can shorten a process. The present invention has been completed by finding a method for preparing a compound of formula (A), an intermediate useful for the preparation thereof, and a method for preparing the compound, which are intended for industrial production to obtain a high purity compound.

According to the present invention, the Edman decomposition can be one time, thereby providing a compound of the formula (A) with high purity and an intermediate useful for its preparation without performing purification by column chromatography in the post-treatment.

That is, this invention relates to the manufacturing method and intermediate shown below.

[1] A method for producing a compound of formula (III) or a salt thereof, wherein the amino acid is released from a compound of formula (II) or a salt thereof in one step.

(Wherein R ^X is -H or a protecting group, iBu is isobutyl, iPr is isopropyl, and Me is methyl)

[2] The compound of the formula (III) obtained by the process described in [1], or a salt thereof, by adding a compound of the formula (I) or a salt thereof to a deprotection reaction to obtain a compound of the formula (II) or a salt thereof A process for preparing a compound of formula (VI) or a salt thereof, characterized by introducing a protecting group into the salt.

Wherein R ^X is -H or a protecting group, iBu is isobutyl, iPr is isopropyl, Me is methyl, Prot ⁰ and Prot ¹ are protecting groups, respectively

[3] The method for producing the compound of the formula (A) or the salt thereof from the compound of the formula (VI) or the salt thereof obtained from the compound of the formula (I) or the salt thereof according to the production method of [2].

Where X is

or

ego,

R ¹ is —H or lower alkyl;

R ² is —H, aryl or lower alkyl,

Wherein the lower alkyl may be substituted with one suitable substituent selected from the group consisting of hydroxy, cycloalkyl, lower alkoxy, aryl, alloalkoxy, substituted carbamoyloxy, and optionally substituted amino, ,

Is nitrogen-containing heterocycle;

Y is

or

ego,

R ³ is cycloalkyl, aryl, optionally substituted heterocyclic or lower alkyl,

Wherein the lower alkyl is selected from the group consisting of hydroxy, cycloalkyl, lower alkoxy, aryl, alloalkoxy, lower alkoxy-lower alkylene-O-, substitutable amino, and -OC (O) NR ⁶ R ⁷ May be substituted by one suitable substituent,

(Wherein R ⁶ and R ⁷ each independently represent —H or lower alkyl or, together with the nitrogen atom to which R ⁶ and R ⁷ bind thereto, represent a nitrogen-containing heterocycle that may be substituted by lower alkyl)

R ⁴ and R ⁵ are each independently —H or lower alkyl;

---- represents a single bond or a double bond

Provided that when R ² is -H, R ³ is cycloalkyl, aryl, substituted heterocycle, lower alkoxymethyl, alloalkyl, t-butyl, sec-butyl, cycloalkyl or ethyl,

Wherein the corresponding ethyl is lower alkyl or hydroxy, lower alkyl-O-, aryl-lower alkylene-O-, lower alkyl-O-lower alkylene-O-, substitutable amino, and -OC (O) NR ⁶ R ⁷ may be substituted by one suitable substituent selected from the group consisting of

[4] Formula (VIII), characterized by condensation of a threonine protected at the carboxylic acid terminus to a compound of the formula (VI) or a salt thereof obtained by the production method of [2], followed by deprotection of the amino terminus. Method for producing the compound or salt thereof.

Wherein Prot ² represents a protecting group

[5] According to the method described in [3], the amino amino acid is condensed at the amino terminus of the compound of formula (VIII) or a salt thereof prepared from the compound of formula (I), and cyclized after deprotection. Process for the preparation of a compound of formula (A) or a salt thereof.

The invention also relates to novel intermediates represented by the formula:

The intermediate obtained by the production method of the present invention is useful in view of the following.

(1) The compound of formula (VIII) obtained in the above-mentioned production method [4] or a salt thereof is useful as an active ingredient of an anti-HCV therapeutic pharmaceutical composition by condensing an amino acid at the N-terminal, deprotecting and then cyclizing it. It is useful as a manufacturing intermediate because it can produce the compound of the formula (A) or a salt thereof which has been reported.

(2) The compound of formula (VI) or a salt thereof obtained in the above-mentioned production method [2] condenses a protected threonine at the C terminus and deprotects the N terminus to thereby prevent the compound of formula (VIII) or a salt thereof. The compound of formula (A) or a salt thereof, which can be prepared, and which has been reported to be useful as an active ingredient of a pharmaceutical composition for anti-HCV treatment according to the method described in (1) above. It is useful as a manufacturing intermediate because it can be prepared.

(3) The compound of formula (III) or a salt thereof obtained in the above-mentioned production method [1] can be prepared by protecting the N terminus after one time of Edman decomposition, to prepare a compound of formula (VI) or a salt thereof, The obtained compound of formula (VI) or a salt thereof according to the method described in (1) and (2) above is a compound of formula (A) or a salt thereof which has been reported to be useful as an active ingredient of a pharmaceutical composition for anti-HCV treatment. Since it can manufacture, it is useful as a manufacturing intermediate.

In addition, unless otherwise indicated, when the symbol in any chemical formula in this specification is used also in another chemical formula, the same symbol shows the same meaning.

The present production method is excellent in operability from the viewpoint that the amino acid can be released in one step, and according to the present production method, for example, a compound of formula (III) or a salt thereof of the present invention may omit the Edman decomposition reaction. In this process, since thiohidantoins and the like are not by-produced, compounds of formula (A), intermediates for their preparation or their salts can be obtained in high purity by crystallization even without purification by chromatography. . Therefore, the scale up on an industrial scale becomes possible.

Hereinafter, the present invention will be described in detail.

In this specification, a "protecting group" is a group which does not interfere with a subsequent reaction and does not affect a functional group even at the time of deprotection. For example, the protecting group described in PGM Wuts and TW Greene, Greene's Protective Groups in Organic Synthesis (4th edition, 2006), etc. are mentioned, According to these reaction conditions, it is appropriate. You can choose to use.

"N-terminal" and "C-terminal" mean an amino group or a carboxy group at both ends in a straight-chain peptide, but "N-terminal" and the side in which a amino group exists are "C-terminal".

"Can be substituted" means having 1 to 5 unsubstituted or substituted groups. In the case of having a plurality of substituents, these substituents may be the same or different from each other.

"Lower" is one, the number of carbon atoms is 1 to 6 unless specifically indicated (hereinafter abbreviated as C _{1 -6),} indicates that preferably one to four.

"Lower alkyl" means a linear or branched C _{1 -6} alkyl, for example methyl, ethyl, n- propyl, isopropyl, n- butyl, isobutyl, sec- butyl, tert- butyl, n- pentyl, n-hexyl and the like. As another embodiment a C _{1 -4} alkyl, a further aspect as the methyl, ethyl, or propyl.

A, such as "lower alkylene" means a straight-chain or branched alkylene group of C _{1 -6} of the ground, for example, methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, propylene, methylmethylene, ethyl ethylene. As another embodiment a C _1-4 alkylene group.

"Lower alkoxy" means methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentyloxy, tert-pentyloxy, neopentyloxy, hexyloxy or isohexyloxy a linear or branched C _{1 -6} alkyl, alkoxy and the like. Another embodiment is methoxy, ethoxy or propoxy.

A, C _{3 -6} alkyl, cyclic alkyl, such as "cycloalkyl" means cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

"Aryl" is phenyl, naphthyl, anthryl and the like.

"Alloalkyl" is lower alkyl substituted with 1 to 5 aryl, such as phenylpropyl, phenethyl, benzyl.

"Aloalkoxy" is lower alkoxy substituted with 1 to 5 aryl, such as phenylpropoxy, phenethyloxy and benzyloxy.

Suitable examples of “substituted amino” are amino, which may be substituted by one or two suitable substituents, such as lower alkyl, amino protecting groups (eg benzyloxycarbonyl, Boc, etc.).

Suitable examples of “substituted carbamoyloxy” are carbamoyloxy which may be substituted by one or two suitable substituents, such as lower alkyl, protecting groups of amino groups (eg benzyloxycarbonyl, Boc, etc.). .

`` Heterocyclic ''

(1) Pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, dihydropyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl (e.g. 4H-1,2,4-tria 1 to 1, such as zolyl, 1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, etc., tetrazolyl (e.g., 1H-tetrazolyl, 2H-tetrazolyl, etc.), azepinyl 3- to 8-membered (more preferably 5- or 6-membered) unsaturated monocyclic heterocycles containing four nitrogen atoms;

(2) 1 to 4 nitrogen atoms such as aziridinyl, azetinyl, pyrrolidinyl, imidazolidinyl, piperidyl, piperazinyl, 2,5-methanopiperazinyl, hexahydroazinyl, and the like. 3 to 8 membered (more preferably 5 to 6 membered) saturated monocyclic heterocycles containing;

(3) indolyl, isoindoleyl, indolinyl, indolinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, tetrahydroin Unsaturated condensed heterocycles containing 1 to 4 nitrogen atoms such as doryl and dihydroindazolyl;

(4) 1, such as oxazolyl, isoxazolyl, oxdiazolyl (e.g. 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.) Or a 3-8 membered (more preferably 5-6 membered) unsaturated monocyclic heterocycle including two oxygen atoms and one to three nitrogen atoms;

(5) a 3 to 8 membered (more preferably 5 to 6 membered) saturated monocyclic heterocyclic ring containing one or two oxygen atoms such as morpholinyl and cydnonyl, and one to three nitrogen atoms;

(6) unsaturated condensed heterocycles containing one or two oxygen atoms, such as benzoxazolyl and benzoxadiazolyl, and 1-3 nitrogen atoms;

(7) thiazolyl, iso thiazolyl, thiadiazolyl (e.g. 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2, 5-thiadiazolyl and the like), 3- to 8-membered (more preferably 5- to 6-membered) unsaturated monocyclic heterocycle including 1 or 2 sulfur atoms such as dihydrothiazinyl, and 1 to 3 nitrogen atoms;

(8) a 3-8 membered (more preferably 5-6 membered) saturated monocyclic heterocycle containing 1 or 2 sulfur atoms such as thiazolidinyl and 1 to 3 nitrogen atoms;

(9) 3-8 membered (more preferably 5-6 membered) unsaturated monocyclic heterocycle containing 1 or 2 sulfur atoms such as thienyl, dihydrodithiyl, dihydrodithionyl, etc .;

(10) unsaturated condensed heterocycles containing one or two sulfur atoms, such as benzothiazolyl, benzothiadiazolyl, imidazothiadiazolyl, and 1-3 nitrogen atoms;

(11) 3 to 8 membered (more preferably 5 to 6 membered) unsaturated monocyclic heterocycle containing one oxygen atom such as furyl;

(12) 3- to 8-membered (more preferably 5- to 6-membered) saturated monocyclic rings containing one or two oxygen atoms, such as oxiranyl, 1,3-dioxolanyl, tetrahydrofuranyl, tetrahydropyranyl, etc. Heterocycle;

(13) 3- to 8-membered (more preferably 5- to 6-membered) unsaturated monocyclic heterocycle including one oxygen atom such as dihydrooxathienyl and one or two sulfur atoms;

(14) unsaturated condensed heterocycles containing one or two sulfur atoms such as benzothienyl and benzodithiinyl;

(15) unsaturated condensed heterocycles containing one oxygen atom and one or two sulfur atoms such as benzooxatiinyl; And

(16) saturated condensed heterocycles containing 1 to 3 nitrogen atoms such as tetrahydropyridopyrrolidinyl; And so on.

Suitable examples of the "nitrogen-containing heterocycle" are (1) to (7) in the "heterocycle" described above, and as another embodiment, at least, such as pyrrolidinyl, piperidyl, morpholinyl, thiazolyl and oxazolyl It is a heterocyclic ring containing one nitrogen atom.

Suitable examples of the "substituted heterocycle" are the aforementioned heterocycles in which one of lower alkyl, lower alkoxy, aryl, amino, lower alkoxycarbonyl and the like can be substituted with an appropriate substituent.

"Halogen" means fluorine, chlorine, bromine and iodine.

In the present specification, the following symbols may be used.

CPME = cyclopentylmethyl ether, DIBOC = di-tert-butyldicarbonate, DME = 1,2-dimethoxyethane, DMF = N, N-dimethylformamide, DMI = 1,3-dimethyl-2-imidazo Dilidinone, DMSO = dimethylsulfoxide, DIPEA = diisopropylethylamine, DPPA = diphenyl phosphide azide, EtOAc = ethyl acetate, Ex = example number, HOBt = 1-hydroxybenzotriazole, IPA = isopropyl Alcohol, KH ₂ PO ₄ = potassium dihydrogen phosphate, NMP = N-methylpyrrolidone, Na ₂ CO ₃ = sodium carbonate, Na ₂ HPO ₄ = sodium hydrogen phosphate, TEA = triethylamine, TFA = trifluoroacetic acid, WSC = 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, tBuOH = tertiarybutanol, LR-ESIMS = low resolution-ESIMS, HR-ESIMS = high resolution-ESIMS, Me = methyl, iBu = isobutyl , iPr = isopropyl, Boc = tertiarybutoxycarbonyl, Ph = phenyl, No. = number.

Geometric isomers may be present in the compounds of the present invention. Although the compound of this invention may be described only in one form of an isomer in this specification, this invention also includes the isomer other than that, and also includes the thing in which the isomer was isolate | separated or a mixture thereof.

In addition, the compound of the present invention has a subtitle carbon atom, and optical isomers based thereon exist. The present invention also includes those in which the optical isomers of the compounds of the present invention are separated or mixtures thereof.

In addition, the salt of the compound of this invention may form a salt with an acid addition salt or a base according to the kind of substituent. Specifically, inorganic acids, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, di Acid addition salts with organic acids such as benzoyltartaric acid, diploylyltartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, glutamic acid, inorganic bases such as sodium, potassium, magnesium, calcium, aluminum, Salts with organic bases such as methylamine, ethylamine, ethanolamine, lysine and ornithine, salts with various amino acids such as acetylleucine and amino acid derivatives, and ammonium salts.

In addition, the present invention also includes various hydrates, solvates, and crystal polymorphic substances of the compound of the present invention and salts thereof. The invention also encompasses compounds labeled with various radioactive or nonradioactive isotopes.

(Production method)

Hereinafter, the typical manufacturing method of the compound of general formula (I)-(VIII) is demonstrated.

(First stage)

Where Prot ⁰ represents a protecting group

This step is a step of deprotecting the N-terminal and then releasing the diamino acid in one step under heating conditions.

When R ^x is a protecting group, lower alkyl which may be substituted with phenyl or lower alkyl which may be substituted with aryl may be mentioned. Other embodiments include lower alkyl, benzyl and tert-butyl.

Examples of Prot ⁰ include carbamate, toluenesulfonyl, nitrobenzenesulfonyl, and the like. As another embodiment, Boc, benzyloxycarbonyl, 9-fluorenylmethyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl and allyloxycarbonyl are mentioned.

Deprotection can be found, for example, in the PGM Wuts and TW Greene, Greene's Protective Groups in Organic Synthesis (4th edition, 2006), see the deprotection reaction for each protecting group. In accordance with these reaction conditions, it may be appropriately selected and used.

The diamino acid is released by heating the raw material intermediate in a solvent. As reaction temperature, Preferably it is a reflux condition from heating. Any solvent may be used as long as it does not interfere with the reaction. For example, CPME, dioxane, THF, DMSO, DMF, IPA, tBuOH, NMP, DMI, DME or water is used, and ether or water is preferable, and CPME- Aqueous systems are particularly preferred. The reaction may be homogeneous or two-layered, but two-layered is particularly preferred. It is sometimes advantageous to carry out the reaction in the presence of an acid or a base or salts. Usually, the reaction solution is added to the reaction solution by adding a buffer solution. Examples of the buffer liquid include a phosphate buffer, a tris buffer, and the like, but a phosphate buffer is preferable. Although the temperature also varies depending on the solvent, the reflux temperature of the solvent is performed at 30 ° C., for example, from 30 ° C. to 180 ° C. 120 degreeC is preferable at 50 degreeC, and 85 degreeC is especially preferable at 60 degreeC. As described in Examples described later, the product may be obtained as a crystal even if no seed crystal is added. However, when the seed crystal is added and the crystal is precipitated, the crystal may be easily precipitated.

(Second stage)

This step is Edman degradation and is a step of leaving the amino acid of the peptide from the N terminus.

At the N-terminus, phenylisothiocyanate is reacted under weakly basic conditions, followed by two steps of leaving the amino acid as 3-phenyl-2-thiohydantoin under acidic conditions. As the solvent, an organic solvent which usually does not adversely affect the reaction, such as alcohols such as acetonitrile, acetone, MeOH, EtOH, THF, dioxane, toluene, methylene chloride, chloroform, EtOAc, DMF, or a mixture thereof It is performed in normal solvents, such as a solvent. The reaction temperature is not limited, and the reaction is usually carried out from cooling to heating. This reaction is described, for example, in M.K.Eberle et al., J. Org. Chem. 59, 7249-7258 (1994)] or the like, or the same method.

(Third step)

Where Prot ¹ and Prot ² both represent protecting groups

This step is a step of protecting the N terminus of an intermediate chain peptide, condensing the protected threonine at the C terminus, and then deprotecting the N terminus.

Examples of Prot ² include lower alkyl which may be substituted with phenyl or lower alkyl which may be substituted with aryl. Other embodiments include lower alkyl, benzyl and tert-butyl.

Examples of Prot ¹ include carbamate, toluenesulfonyl, nitrobenzenesulfonyl, and the like. As another embodiment, Boc, benzyloxycarbonyl, 9-fluorenylmethyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl and allyloxycarbonyl are mentioned.

In order to condense the protected threonine to the C terminal of the chain peptide of the substrate, it can be carried out in the same manner as the so-called amidation reaction. Specifically, the compound of formula (VII) or a salt thereof can be obtained by reaction of a compound of formula (VI) or a salt thereof with a protected threonine of formula (B2). In this reaction, the compound of formula (VI) or a salt thereof and the protected threonine of formula (B2) are used in an equal or excess amount, and the mixture is cooled in a solvent inert to the reaction, in the presence of a condensing agent, under cooling. Under heating, stirring is preferably performed at -20 ° C to 60 ° C, usually for 0.1 hours to 5 days. Examples of the solvent used herein are not particularly limited, but aromatic hydrocarbons such as toluene or xylene, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane or chloroform, diethyl ether, THF, dioxane and dimethoxyethane Ethers such as DMF, DMSO, EtOAc, MeCN or water, and mixtures thereof. Examples of the condensing agent include, but are not limited to, WSC, DPPA, and phosphorus oxychloride. It is sometimes preferable to use an additive (for example, HOBt) for the reaction. It is sometimes advantageous to carry out the reaction in the presence of an organic base such as TEA, DIPEA or N-methylmorpholine, or an inorganic base such as K ₂ CO ₃ , Na ₂ CO ₃ or KOH.

It is also possible to use a method of converting a compound of formula (VII) or a salt thereof to a reactive derivative followed by reaction with a protected threonine of formula (B2). As an example of the reactive derivative of carboxylic acid, the acid halide obtained by reaction with halogenating agents, such as phosphorus oxy chloride and thionyl chloride, the mixed acid anhydride obtained by reaction with isobutyl chloroformate, etc., the active ester obtained by condensation with HOBt etc., etc. are mentioned. Can be. The reaction of these reactive derivatives with the protected threonine of formula (B2) is carried out in a solvent inert to the reaction of halogenated hydrocarbons, aromatic hydrocarbons, ethers and the like, under heating under cooling, preferably at -20 ° C to 60 ° C. I can do it.

SR Sandler and W. Karo, Organic Functional Group Preparations, 2nd edition, Vol. 1, Academic Press Inc., 1991 Japanese Chemical Society, `` Experimental Chemistry Lectures (Fifth Edition), Vol 16 ( 2005) (Maruzen)

(Fourth step)

Compounds of formula (III) or salts thereof, compounds of formula (V) or salts thereof, or compounds of formula (VIII) or salts thereof are protected or deprotected by appropriate processes, using appropriate protecting groups, and By condensation, the compound of formula (A) or a salt thereof can be prepared. Since the cyclization reaction is amidation, it can be carried out by the same method as the above-mentioned third step, and therefore, the reagent and reaction conditions (for example, solvent, reaction temperature, etc.) used can refer to the above-mentioned third step.

(Raw material synthesis)

As above-mentioned, the compound of general formula (I) or its salt can be manufactured according to patent document 1 using FR901459 substance as a starting material.

FR901459 materials include Stachybotrys chartarum No. 19292, International Patent Organism Depositary, Postal Code 305-8566 1-1-1 Higashi, Tsukuba City, Ibaragi Prefecture Chuo No. 6, Tokuri It can be obtained, for example, by the method described in Patent Literature 2, from a culture solution of Tsu Kosei Hojin Sangyo Kijutsu Sogo Genshusho, deposited internationally on April 16, 1991 as accession number FERM BP-3364.

Example compounds are isolated and purified as free compounds, their salts, hydrates, solvates or crystalline polymorphs. EXAMPLES The salt of a compound can also be manufactured by adding to the salt-forming reaction of a conventional method. Isolation and purification are performed by applying extraction and fractional crystallization.

<Examples>

Hereinafter, the preparation method of the compound of general formula (I)-(VIII) is demonstrated in detail based on an Example. Example 4 The structure of the compound is shown in Table 4.

Example 1 Preparation of Compounds of Formula (I), wherein Prot ⁰ is Boc

According to the method of patent document 1, (6S, 12S, 15S, 18S, 21S, 24R, 27S, 30S, 33S, 36S) -6-[(1R) -1-hydroxyethyl] -36-[(1R , 2R, 4E) -1-hydroxy-2-methyl-4-hexen-1-yl] -12,15,18,27,30-pentisobutyl-33-isopropyl-2,2,8,11 , 17,21,24,26,32,35-decamethyl-4,7,10,13,16,19,22,25,28,31,34-undecaoxo-3-oxa-5,8, 93.8 g of 11,14,17,20,23,26,29,32,35-undecazaheptatricontan-37-oic acid were obtained.

Example 2 (Preparation of Compound of Formula (II))

After dissolving 50.0 g of Example Compound 1 in 500 mL of methylene chloride, the mixture was cooled to 5 ° C, and 213.2 g of TFA was added dropwise and stirred for 4 hours. Thereafter, 500 mL of cold water was added, and pH was adjusted to 6.5 with 10 (w / v)% Na ₂ CO ₃ aqueous solution. After separating, 500 mL of cooled water was added again to the organic layer, and pH was adjusted to 6.5-7 with 10 (w / v)% Na ₂ CO ₃ aqueous solution. After separation, the organic layer was separated, and again and again, 500 mL of cooled water was added, and pH was adjusted to 6.5-7 with 10 (w / v)% Na ₂ CO ₃ aqueous solution. After separating, the lower layer was concentrated to about 250 mL, 500 mL of CPME was added, and again concentrated to about 250 mL. 500 mL of CPME was added and concentrated under reduced pressure to about 250 mL. Example 250 mL of a CPME solution of compound 2 was obtained.

Example 3 (Preparation of Compound of Formula (III))

Example In about 250 mL of a CPME solution of compound 2, 500 mL of a phosphate buffered solution of 500 mL of CPME, pH 7.6 (the pH of Na ₂ HPO ₄ aqueous solution was prepared to pH 7.6 with KH ₂ PO ₄ aqueous solution, and Na ₂ HPO ₄ aqueous solution). Was prepared by dissolving 6.26 g of Na ₂ HPO ₄ .12H ₂ O in 522 mL of silver water, and an aqueous KH ₂ PO ₄ solution was prepared by dissolving 0.377 g of KH ₂ PO ₄ in 81 mL of water). It heated up at 80-85 degreeC, stirred for 5 hours, and cooled to 25 degreeC. The organic layer obtained by separating off was washed twice with 500 mL of water. The organic layer was concentrated to about 105 mL, and then 115 mL of acetonitrile was added dropwise. Then, 75 mg of seed crystals of Example compound 3 were added at 22 degreeC internal temperature, and 345 mL of acetonitrile was further dripped. The crystallized solution was filtered off and dried under reduced pressure to obtain (2S, 3R, 4R, 6E) -3-hydroxy-4-methyl-2- [methyl (N-methyl-L-leucine-L-leucil-N-methyl). -L-leucil-L-alanyl-D-alanyl-N-methyl-L-leucil-L-leucil-N-methyl-L-valyl) amino] oct-6-enoic acid was obtained as a white powder. Yield: 22.7 g, yield: 56.9%

In addition, a part of the reaction solution of Example 3 was collected, and it was confirmed that 3- (1-hydroxyethyl) -1-methylpiperazine-2,5-dione was produced by the following analysis results.

<Example 3-1>

(Method for producing amorphous)

Example In a CPME solution of Compound 2 (equivalent to 40.0 g / 800 mL), 400 mL of an aqueous solution of pH 7.6 phosphate buffer (a pH of an aqueous solution of Na ₂ HPO ₄ was adjusted to pH 7.6 with an aqueous KH ₂ PO ₄ solution, and an aqueous solution of Na ₂ HPO _4). Was prepared by dissolving 6.26 g of Na ₂ HPO ₄ .12H ₂ O in 522 mL of silver water, and aqueous KH ₂ PO ₄ solution was prepared by dissolving 0.377 g of KH ₂ PO ₄ in 81 mL of water). It heated up at 85 degreeC, stirred for 5 hours and 30 minutes, and cooled to room temperature. The organic layer obtained by separating off was washed sequentially with 400 mL of time water and 400 mL of 20 (w / v)% saline. The organic layer was concentrated to about 280 mL and then added dropwise in 1800 mL n-heptane. The precipitated powder was filtered out and dried under reduced pressure, followed by (2S, 3R, 4R, 6E) -3-hydroxy-4-methyl-2- [methyl (N-methyl-L-leucine-L-leucine-N-). Methyl-L-leucil-L-alanyl-D-alanyl-N-methyl-L-leucil-L-leucil-N-methyl-L-valyl) amino] oct-6-enoic acid was obtained as a powder. Yield: 32.17 g, yield: 93.4%

(Production method of seed crystals)

In a mixed solvent of 40 mL of CPME and 160 mL of acetonitrile, 25.0 g of the powder of Example Compound 3 was dissolved at 25-45 ° C. Then, it cooled slowly to 20 degreeC over 1 hour, and stirred at 20 degreeC for 18 hours. The precipitated crystals were filtered off and dried under reduced pressure to obtain (2S, 3R, 4R, 6E) -3-hydroxy-4-methyl-2- [methyl (N-methyl-L-leucine-L-leucil-N-). Methyl-L-leucil-L-alanyl-D-alanyl-N-methyl-L-leucil-L-leucil-N-methyl-L-valyl) amino] oct-6-enoic acid was obtained as crystals. Yield: 13.02 g, yield: 52.1%

Example 4 (Preparation of Compound of Formula (IV))

After dissolving 10.0 g of Example Compound 3 in 150 mL of EtOAc, 50 mL of pH 7.6 phosphate buffer (the pH of Na ₂ HPO ₄ aqueous solution was adjusted to pH 7.6 with KH ₂ PO ₄ aqueous solution, and the Na ₂ HPO ₄ aqueous solution was Prepared by dissolving Na ₂ HPO ₄ .12H ₂ O, 1.04 g in 87.5 L of water, and aqueous KH ₂ PO ₄ solution prepared by dissolving 0.568 g of KH ₂ PO ₄ in 12.5 L of water), phenylisothiocyanate 2.54 g was added and stirred for 4 hours. Thereafter, the pH was adjusted to 2.3 with 1 M hydrochloric acid. After separation, the organic layer was aliquoted, washed with 100 mL of 20 (w / v)% brine, and concentrated to about 40 mL. Example 40 mL of an EtOAc solution of compound 4 was obtained.

Example 5 (Preparation of Compound of Formula (V))

Example To about 40 mL of an EtOAc solution of compound 4, further dropwise in 200 mL of n-heptane. The precipitated powder was collected by filtration, dissolved in 100 mL of acetonitrile, 94 mL of 1M hydrochloric acid was added, and the mixture was stirred at 20 ° C. for 3 hours and 15 minutes. Example Compound 5 was obtained.

Example 6 Preparation of Compounds of Formula (VI), wherein Prot ¹ is Boc

Example 10 (w / v)% Na ₂ CO ₃ aqueous solution was further added to the reaction solution of compound 5, the pH was adjusted to about 7, DIBOC 2.25 g was added, and the mixture was stirred at 25 ° C. for 3 hours 20 minutes. The pH was then adjusted to 2.5 with 1 M hydrochloric acid. The organic layer was separated, 100 mL of 20 (w / v)% saline was added, and then extracted with 100 mL of EtOAc. The organic layer was separated, concentrated to about 40 mL, and again 150 mL of EtOAc, concentrated to about 40 mL. 150 mL of EtOAc was added again and again to this concentrate, it concentrated to about 40 mL, it cooled to 5 degreeC, and it dripped in 40 mL of n-heptane previously cooled to 5 degreeC. The precipitated powder was filtered and then dried under reduced pressure to obtain (2S, 3R, 4R, 6E) -2-{[N- (tert-butoxycarbonyl) -L-leucine-N-methyl-L-leucine-L- Alanyl-D-alanyl-N-methyl-L-leucil-L-leucil-N-methyl-L-valyl] (methyl) amino} -3-hydroxy-4-methyloct-6-enoic acid white powder Obtained as. Yield: 7.6 g, yield: 77.9%

Example 7 Preparation of Compounds of Formula (VII), wherein Prot ¹ is Boc and Prot ² is Methyl

7.5 g of Example compound 6 was dissolved in 75 mL of methylene chloride, 1.47 g of L-threoninemethylester hydrochloride and 1.47 g of HOBt. Monohydrate were added, and the mixture was cooled to 0 to 10 ° C. Further, 1.12 g of WSC was added, the mixture was stirred at 5 ° C. for about 20 hours, and then the organic layer was washed with 75 mL of water. The aqueous layer was reextracted with 38 mL of methylene chloride and the organic layers were combined. The organic layer was washed with 68 mL of 0.3 M hydrochloric acid, and the aqueous layer was again extracted with 38 mL of methylene chloride. The organic layers were combined, washed sequentially with 76 mL of 5 (w / v)% NaHCO ₃ aqueous solution and 55 mL of 20 (w / v)% brine, and concentrated to about 30 mL. The concentrate was added dropwise in 300 mL of heptane. The precipitated powder was filtered out and dried under reduced pressure to obtain (N-[(2S, 3R, 4R, 6E) -2-{[N- (tert-butoxycarbonyl) -L-leucine-N-methyl- L-leucil-L-alanyl-D-alanyl-N-methyl-L-leucil-L-leucil-N-methyl-L-valyl] (methyl) amino} -3-hydroxy-4-methyloct- 6-Enoyl] -L-threoninemethylester was obtained as a white powder Yield: 7.12 g, yield: 85.5%

Example 8 Preparation of Compounds wherein Prot ² Is Methyl In Formula (VIII)

20.0 g of Example compound 7, were dissolved in 120 mL of methylene chloride and cooled to -5 ° C. 98.9 g of TFA was added dropwise, stirred for 2 hours and 40 minutes, and then washed with cooled 200 hours of water. The organic layer obtained by separating off was washed with 200 mL of further cooled water. 200 mL of cooled water was added to the organic layer, and pH was adjusted to 6.5-7 using 10 (w / v)% Na ₂ CO ₃ aqueous solution. A separating the resulting organic layer, was prepared again was added to 200 mL can naengsi, and 10 to a pH of _{(w / v)% Na 2} CO 3 aqueous solution at 6.5 to 7. Aliquot was added to again be repeated naengsi 200 mL of the obtained organic layer, which was prepared to a pH of 10 (w / v)% Na 2 CO 3 aqueous solution at 6.5 to 7. The organic layer obtained by separating off was concentrated under reduced pressure to about 50 mL. The concentrated solution was added dropwise in 1500 mL of heptane, and the precipitated powder was filtered out and dried under reduced pressure to obtain N-{(2S, 3R, 4R, 6E) -3-hydroxy-2-[(L-leucine-N -Methyl-L-leucil-L-alanyl-D-alanyl-N-methyl-L-leusil-L-leucil-N-methyl-L-valyl) (methyl) amino] -4-methyloct-6- Enoyl} -L-threoninemethylester was obtained as a white powder. Yield: 16.82 g, yield: 92.1%

The obtained Example compound 8 compared with the compound of Prep3 of patent document 1, and NMR and MS data, and confirmed that it was the same structure.

The present production method is excellent in operability from the viewpoint of leaving off an amino acid in one step, and according to the present production method, for example, the compound of formula (III) or a salt thereof of the present invention omits the Edman decomposition reaction. In the process which can be used, since thiohydrantoin etc. are not by-produced, the compound of Formula (A), the intermediate for its preparation, or each salt thereof can be obtained in high purity by crystallization even without purification by chromatography. have. Therefore, the scale up on an industrial scale becomes possible.

Microbiological industry research institute BP-3364 19910416 Microbiological industry research institute BP-3364 19910615

Claims (6)

A process for producing a compound of formula (III) or a salt thereof, wherein the amino acid is released from a compound of formula (II) or a salt thereof in one step.

(Wherein R ^X is -H or a protecting group, iBu is isobutyl, iPr is isopropyl, and Me is methyl) A compound of formula (I) or a salt thereof is added to the deprotection reaction to obtain a compound of formula (II) or a salt thereof, and also a protecting group to a compound of formula (III) or a salt thereof obtained by the process according to claim 1. A process for producing a compound of formula (VI) or a salt thereof, characterized by the introduction of

Wherein R ^X is -H or a protecting group, iBu is isobutyl, iPr is isopropyl, Me is methyl, Prot ⁰ and Prot ¹ are protecting groups, respectively A process for producing a compound of formula (A) or a salt thereof, according to the process of claim 2, from a compound of formula (VI) or a salt thereof obtained from a compound of formula (I) or a salt thereof.

Where X is

or

ego,
R ¹ is —H or lower alkyl;
R ² is —H, aryl or lower alkyl,
Wherein the lower alkyl may be substituted with one suitable substituent selected from the group consisting of hydroxy, cycloalkyl, lower alkoxy, aryl, alloalkoxy, substituted carbamoyloxy, and optionally substituted amino, ,

Is nitrogen-containing heterocycle;
Y is

or

ego,
R ³ is cycloalkyl, aryl, optionally substituted heterocyclic or lower alkyl,
Wherein the lower alkyl is selected from the group consisting of hydroxy, cycloalkyl, lower alkoxy, aryl, alloalkoxy, lower alkoxy-lower alkylene-O-, substitutable amino, and -OC (O) NR ⁶ R ⁷ May be substituted by one suitable substituent,
(Wherein R ⁶ and R ⁷ each independently represent —H or lower alkyl or, together with the nitrogen atom to which R ⁶ and R ⁷ bind thereto, represent a nitrogen-containing heterocycle that may be substituted by lower alkyl)
R ⁴ and R ⁵ are each independently —H or lower alkyl;
---- represents a single bond or a double bond
Provided that when R ² is -H, R ³ is cycloalkyl, aryl, substituted heterocycle, lower alkoxymethyl, alloalkyl, t-butyl, sec-butyl, cycloalkyl or ethyl,
Wherein the corresponding ethyl is lower alkyl or hydroxy, lower alkyl-O-, aryl-lower alkylene-O-, lower alkyl-O-lower alkylene-O-, substitutable amino, and -OC (O) NR ⁶ R ⁷ may be substituted by one suitable substituent selected from the group consisting of According to the preparation method of claim 2, the compound of formula (VI) obtained from the compound of formula (I) or a salt thereof or a salt thereof is condensed with a threonine protected at the carboxylic acid end, and the amino terminus is deprotected. Method for producing a compound of formula (VIII) or a salt thereof.

(Wherein iBu isobutyl, iPr isopropyl, Me is methyl and Prot ² is a protecting group) According to the method according to claim 3, the amino acid is condensed at the amino terminus of a compound of formula (VIII) or a salt thereof prepared from a compound of formula (I) or a salt thereof and cyclized after deprotection. Process for the preparation of the compound of A) or a salt thereof. The compound selected from the group which consists of the following, or its salt.
TABLE 3

(Wherein iBu isobutyl, iPr isopropyl, Boc tertiarybutoxycarbonyl, Me is methyl and Ph is phenyl)