WO2019176732A1

WO2019176732A1 - Production method for tetrahydroisoquinoline ring-containing compound

Info

Publication number: WO2019176732A1
Application number: PCT/JP2019/009128
Authority: WO
Inventors: 博毅大栗; 涼谷藤; 及川　英秋
Original assignee: 国立大学法人東京農工大学; 国立大学法人北海道大学
Priority date: 2018-03-16
Filing date: 2019-03-07
Publication date: 2019-09-19
Also published as: JPWO2019176732A1

Abstract

[Problem] To provide a production method capable of easily synthesizing a compound having a structure in which a plurality of tetrahydroisoquinoline rings are linked together, using a reduced number of steps. [Solution] It was found that a non-naturally-occurring intermediate compound having a pentacyclic backbone in which tetrahydroisoquinoline rings are linked together, can be quickly one-pot synthesized by reacting a tyrosine derivative with a non-naturally-occurring aldehyde substrate having a particular structure by using a nonribosomal peptide synthetase. Also, it was found that saframycins and analogs thereof can be easily and highly efficiently obtained using a non-naturally-occurring aldehyde substrate having a particular structure, where adaptiveness to an enzyme reaction is ensured, and chemical transformation of the intermediate after formation of the backbone can be freely performed.

Description

Method for producing tetrahydroisoquinoline ring-containing compound

The present invention relates to a method for producing a compound having a structure in which a plurality of tetrahydroisoquinoline rings are linked, and particularly to a method for producing saframycins and analogs thereof. The present invention also relates to an intermediate compound for synthesizing the compound.

The antitumor alkaloid group represented by saframycins is a compound having a complex pentacyclic skeleton in which a plurality of tetrahydroisoquinoline (THIQ) rings are linked. For example, saframycin A (compound 1) and saframycin Y3 (compound 2), which are saframycins, have the structures shown below. In addition, many analogs such as Jornamycin A, Renieramycin M, Extenasaidin 743 (Compounds 3 to 5) have been reported as a group of natural products sharing a similar pentacyclic mother skeleton. Compounds 1 to 4 are composed of two THIQ rings and have a function of alkylating a guanine base of DNA with an iminium cation generated from an aminonitrile. Further, Compound 5 having both a nucleic acid alkylation site and a protein interaction site has been clinically applied as an anticancer drug ("Trabectedin", "Yonderis").

For the tetrahydroisoquinoline alkaloid compound group having such a unique structure and exhibiting excellent anticancer activity, various total synthesis studies have been actively developed (Non-patent Documents 1 and 2). In addition, the above-mentioned ectenasaidin 743 (compound 5), which is an anticancer drug with a very complex polycyclic structure, is supplied in a multi-stage semi-synthetic cocoon (21 steps) cocoon obtained from cyanosafracin (compound 6) obtained by culture. (Non-Patent Document 3). However, these conventionally reported synthesis processes involve complicated starting materials, and it is necessary to separate and purify the product every time the reaction is performed, so that there is a problem of production efficiency that requires a multi-step synthesis step. was there.

On the other hand, to date, the inventors have not promoted peptide formation, which is the original function of SfmC module protein, which is a non-ribosome-dependent peptide synthase (NRPS) that catalyzes biosynthesis of saframycin A, An enzyme with unique properties that forms a pentacyclic skeleton containing a THIQ ring by catalyzing imine formation between long-chain fatty acid aldehydes and amino acids and subsequent Pictet-Spengler (PS) -type cyclization This is reported (Non-patent Document 4). However, in the situation where there are many other functional groups in the molecule, it has been extremely difficult to selectively remove the long-chain fatty acid moiety essential for such enzymatic reaction. Therefore, after forming a pentacyclic skeleton using an enzymatic reaction, there has been a limit to the chemical synthesis of the desired saframycins and novel analogs by chemical modification.

From such a background, an object of the present invention is to provide a production method capable of easily synthesizing a compound having a structure in which a plurality of tetrahydroisoquinoline rings are linked with fewer steps. It is another object of the present invention to provide a chemically modifiable intermediate that enables the synthesis of novel derivatives other than natural tetrahydroisoquinoline ring-containing alkaloids.

As a result of intensive studies to solve the above-mentioned problems, the present inventors have used a non-natural aldehyde substrate and a tyrosine derivative having a specific structure that do not exist in the biosynthetic pathway using a non-ribosome-dependent peptide synthase. It was found that a non-natural intermediate compound having a pentacyclic skeleton linked with a tetrahydroisoquinoline ring can be rapidly synthesized in one pot. In addition, by using a non-natural aldehyde substrate having a specific structure, chemical conversion after skeleton formation of an intermediate can be freely performed while ensuring adaptability to an enzyme reaction, and it is simple and highly efficient. It has been found that a class and its analogs are obtained. Based on these findings, the present invention has been completed.

That is, the present invention in one aspect,
<1> A method for producing a compound represented by the following formula (I):

Wherein X is —C (R ³ ) NH— or —C (R ³ ) O—; Y is —C (═O) R ⁴ —NH—, —C (═O) R ⁴ —O—, —S (═O) ₂ R ⁴ —NH—, —S (═O) ₂ R ⁴ —O—, —P (═O) (OR ⁵ ) R ⁴ —NH—, or —P (═O) (OR ⁵ ) R ⁴ —O—; Z is —C (═O) R ⁶ , —C (═O) OR ⁶ , —S (═O) ₂ R ⁶ , or —P (═O) (OR ⁵ ) R ⁶ ; R ¹ and R ² are each independently a C ₁ -C ₁₀ alkyl group, which may be the same or different; R ³ is a hydrogen atom or a substituent An optionally substituted C ₁ -C ₅ alkyl group; R ⁴ is an optionally substituted alkylene group; R ⁵ is an optionally substituted C ₁ -C ₅ alkyl group; R ⁶ is a saturated or A hydrocarbon chain, saturated; R ^a is optionally substituted alkyl group).
[Step A] a compound represented by the following formula (a):

(Wherein R ¹ and R ² have the same meaning as in formula (I).)
A compound represented by the following formula (b):

(In the formula, X, Y and Z have the same meaning as in formula (I).)
A step of obtaining a compound represented by the following formula (II) by reacting in the presence of a non-ribosome-dependent peptide synthase

(Wherein X, Y, Z, R ¹ and R ² have the same meaning as in formula (I));
[Step B] A step of adding a cyanide ion to the compound represented by the formula (II) to obtain a compound represented by the following formula (III)

(Wherein X, Y, Z, R ¹ and R ² have the same meaning as in formula (I)); and [Step C] an aldehyde and a reducing agent for the compound represented by formula (III) N-alkylation of a secondary amine moiety by adding a compound to obtain a compound represented by the formula (I);
<2> The production method according to <1>, wherein the non-ribosome-dependent peptide synthase is saframycin A biosynthetic enzyme SfmC;
<3> The production method according to <1> or <2> above, which comprises adding ATP, NADPH, Mg ²⁺ , and Mn ²⁺ in Step A;
<4> The production method according to any one of <1> to <3>, wherein R ¹ and R ² are both methyl groups; and <5> the compound represented by formula (b): The production method according to any one of the above <1> to <4>, selected from the group consisting of the following compounds;

(Wherein, ^{Z a} is a C ₆ -C ₂₀ alkyl group, Me represents a methyl group.)
Is to provide.

In one preferred embodiment, the present invention provides
<6> From an intermediate compound represented by the following formula (I)

Wherein X is —C (R ³ ) NH—; Y is —C (═O) R ⁴ —NH—; Z is —C (═O) OR ⁶ ; R ¹ and R ² are each independently a C ₁ -C ₁₀ alkyl group, which may be the same or different; R ³ is a hydrogen atom or an optionally substituted C ₁ -C ₅ alkyl group R ⁴ is an optionally substituted alkylene group; R ⁶ is a saturated or unsaturated hydrocarbon chain; R ^a is an optionally substituted alkyl group.)
A method for producing a compound represented by the following formula (IV):

(Wherein X, Y, R ¹ , R ² and R ^a have the same meaning as in formula (I), and A is a hydrogen atom or a protecting group);
[Step D] A step of obtaining a compound represented by the following formula (V) by converting the Z site of the compound represented by the formula (I) into A in the presence of a transition metal catalyst.

(In the formula, X, Y, A, R ¹ , R ² and R ^a have the same meaning as in formula (IV).)
[Step E] The production comprising the step of obtaining the compound represented by the formula (IV) by oxidizing the compound represented by the formula (V) in the presence of a transition metal catalyst and converting phenol to quinone. Method;
^<7> R 1, ^{R 2} and ^{R a} are both a methyl group; ^{R 3} is hydrogen atom; ^{R 4} is, -CH (CH ₃₎ - is, according to the above <6> Production method;
<8> The method according to <6> or <7> above, wherein the transition metal catalyst in Step D is a palladium complex; and <9> the above <6, wherein the transition metal catalyst in Step E is a cobalt complex. The manufacturing method according to any one of <8> to <8> is provided.

In another preferred embodiment, the present invention provides:
<10> From the intermediate compound represented by the following formula (I)

Wherein X is —C (R ³ ) NH—; Y is —C (═O) R ⁴ —O—; Z is —C (═O) R ⁶ ; R ¹ and R ² are each independently a C ₁ -C ₁₀ alkyl group, which may be the same or different; R ³ is a hydrogen atom or an optionally substituted C ₁ -C ₅ alkyl group R ⁴ is a 1-methylmethylene group; R ⁶ is a saturated or unsaturated hydrocarbon chain; R ^a is an optionally substituted alkyl group.)
A method for producing a compound represented by the following formula (VI):

(Wherein X, R ¹ , R ² and R ^a have the same meaning as in formula (I));
[Step F] A compound represented by the following formula (VII) is obtained by hydrolyzing the compound represented by the above formula (I) under basic conditions and converting the end of the Y site into an OH group. Process

(In the formula, X, R ¹ , R ² and R ^a have the same meaning as in formula (I).)
[Step G] A step of obtaining a compound represented by the following formula (VIII) by oxidizing the compound represented by the formula (VII) in the presence of a transition metal catalyst and converting phenol to quinone.

(Wherein X, R ¹ , R ² and R ^a have the same meaning as in formula (I)); and [Step H] The compound represented by (VIII) is subjected to DMSO oxidation or hypervalence. The production method comprising a step of obtaining a compound represented by the formula (VI) by converting a terminal OH group into a carbonyl group by oxidation with an iodine reagent;
<11> The production method according to the above <10>, wherein R ¹ , R ² and R ^a are all methyl groups; and R ³ is —CH (CH ₃ ) —; and <12> Step G The transition metal catalyst in is a cobalt complex, The manufacturing method as described in said <10> or <11> is provided.

In one preferred embodiment, the present invention also relates to the following intermediate compounds: In particular,
<13> A compound represented by the following formula (I);

Wherein X is —C (R ³ ) NH— or —C (R ³ ) O—; Y is —C (═O) R ⁴ —NH—, —C (═O) R ⁴ —O—, —S (═O) ₂ R ⁴ —NH—, —S (═O) ₂ R ⁴ —O—, —P (═O) (OR ⁵ ) R ⁴ —NH—, or —P (═O) (OR ⁵ ) R ⁴ —O—; Z is —C (═O) R ⁶ , —C (═O) OR ⁶ , —S (═O) ₂ R ⁶ , or —P (═O) (OR ⁵ ) R ⁶ ; R ¹ and R ² are each independently a C ₁ -C ₁₀ alkyl group, which may be the same or different; R ³ is a hydrogen atom or a substituent An optionally substituted C ₁ -C ₅ alkyl group; R ⁴ is an optionally substituted alkylene group; R ⁵ is an optionally substituted C ₁ -C ₅ alkyl group; R ⁶ is a saturated or A hydrocarbon chain, saturated; R ^a is optionally substituted alkyl group).
<14> The compound according to <13>, selected from the group consisting of the following compounds;

(Wherein, ^{Z a} _is a C 6 _{-C 20} alkyl group, Me represents a methyl group.)
<15> a compound represented by the following formula (II);

Wherein X is —C (R ³ ) NH— or —C (R ³ ) O—; Y is —C (═O) R ⁴ —NH—, —C (═O) R ⁴ —O—, —S (═O) ₂ R ⁴ —NH—, —S (═O) ₂ R ⁴ —O—, —P (═O) (OR ⁵ ) R ⁴ —NH—, or —P (═O) (OR ⁵ ) R ⁴ —O—; Z is —C (═O) R ⁶ , —C (═O) OR ⁶ , —S (═O) ₂ R ⁶ , or —P (═O) (OR ⁵ ) R ⁶ ; R ¹ and R ² are each independently a C ₁ -C ₁₀ alkyl group, which may be the same or different; R ³ is a hydrogen atom or a substituent An optionally substituted C ₁ -C ₅ alkyl group; R ⁴ is an optionally substituted alkylene group; R ⁵ is an optionally substituted C ₁ -C ₅ alkyl group; R ⁶ is a saturated or A hydrocarbon chain, saturated; R ^a is optionally substituted alkyl group).
<16> The compound according to <15>, which has the following structure;

(Wherein, ^{Z a} _is a C 6 _{-C 20} alkyl group, Me represents a methyl group.)
<17> an aldehyde compound represented by the following formula (b);

Wherein X is —C (R ³ ) NH— or —C (R ³ ) O—; Y is —C (═O) R ⁴ —NH—, —C (═O) R ⁴ —O—, —S (═O) ₂ R ⁴ —NH—, —S (═O) ₂ R ⁴ —O—, —P (═O) (OR ⁵ ) R ⁴ —NH—, or —P (═O) (OR ⁵ ) R ⁴ —O—; Z is —C (═O) R ⁶ , —C (═O) OR ⁶ , —S (═O) ₂ R ⁶ , or —P (═O) (OR ⁵ ) R ⁶ ; R ³ is a hydrogen atom or an optionally substituted C ₁ -C ₅ alkyl group; R ⁴ is an optionally substituted alkylene group R ⁵ is an optionally substituted C ₁ -C ₅ alkyl group; R ⁶ is a saturated or unsaturated hydrocarbon chain.)
<18> The aldehyde compound according to <17>, selected from the group consisting of the following compounds.

(Wherein, ^{Z a} _is a C 6 _{-C 20} alkyl group, Me represents a methyl group.)
Is to provide.

According to the production method of the present invention, a non-natural intermediate compound having a pentacyclic skeleton linked by a tetrahydroisoquinoline ring can be rapidly synthesized in one pot. By using a synthesis scheme through such an intermediate compound, the terminal long-chain fatty acid moiety can be selectively removed even after the formation of the pentacyclic skeleton even in the presence of many other functional groups, compared with conventional synthesis methods. Thus, an excellent effect is obtained that saframycins and analogs thereof can be efficiently obtained in a short process.

In addition, the intermediate compound of the present invention can not only selectively remove the long-chain fatty acid moiety, which has been difficult in the conventional enzyme-catalyzed reaction as described above, but also can be substituted with any substituent after the intermediate skeleton is formed. Since it can be introduced and subjected to chemical conversion freely, it can be applied to the synthesis of novel derivatives other than natural tetrahydroisoquinoline ring-containing alkaloids. Furthermore, there is an advantage that benzene rings / quinone rings at both ends of the pentacyclic skeleton involved in the activity expression of the final target product can be separately produced.

Hereinafter, embodiments of the present invention will be described. The scope of the present invention is not limited to these descriptions, and other than the following examples, the scope of the present invention can be appropriately changed and implemented without departing from the spirit of the present invention.

1. Definitions In the present specification, the “alkyl or alkyl group” may be any of an aliphatic hydrocarbon group composed of a straight chain, a branched chain, a ring, or a combination thereof. The number of carbon atoms of the alkyl group is not particularly limited. For example, the number of carbon atoms is 1 to 20 (C _1-20 ), the number of carbons is 1 to 15 (C _{1 to 15} ), and the number of carbon atoms is 1 to 10 (C _{1 to 10).} ). In the present specification, the alkyl group may have one or more arbitrary substituents. For example, C _1-8 alkyl includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neo-pentyl, n-hexyl, isohexyl, n-heptyl, n-octyl and the like are included. Examples of the substituent include an alkoxy group, a halogen atom (which may be a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom), an amino group, a mono- or di-substituted amino group, a substituted silyl group, or Although acyl etc. can be mentioned, it is not limited to these. When the alkyl group has two or more substituents, they may be the same or different. The same applies to the alkyl part of other substituents containing an alkyl part (for example, an alkoxy group, an arylalkyl group, etc.).

In the present specification, “alkylene” is a divalent group consisting of a linear or branched saturated hydrocarbon, such as methylene, 1-methylmethylene, 1,1-dimethylmethylene, ethylene, 1-methylethylene, 1-ethylethylene, 1,1-dimethylethylene, 1,2-dimethylethylene, 1,1-diethylethylene, 1,2-diethylethylene, 1-ethyl-2-methylethylene, trimethylene, 1 -Methyltrimethylene, 2-methyltrimethylene, 1,1-dimethyltrimethylene, 1,2-dimethyltrimethylene, 2,2-dimethyltrimethylene, 1-ethyltrimethylene, 2-ethyltrimethylene, 1,1 -Diethyltrimethylene, 1,2-diethyltrimethylene, 2,2-diethyltrimethylene, 2-ethyl-2-methyltrime Len, tetramethylene, 1-methyltetramethylene, 2-methyltetramethylene, 1,1-dimethyltetramethylene, 1,2-dimethyltetramethylene, 2,2-dimethyltetramethylene, 2,2-di-n-propyl Trimethylene and the like.

In the present specification, when a functional group is defined as “may be substituted”, the type of substituent, the position of substitution, and the number of substituents are not particularly limited, and two or more When they have a substituent, they may be the same or different. Examples of the substituent group include, but are not limited to, an alkyl group, an alkoxy group, a hydroxyl group, a carboxyl group, a halogen atom, a sulfo group, an amino group, an alkoxycarbonyl group, and an oxo group. These substituents may further have a substituent. Examples of such include, but are not limited to, a halogenated alkyl group.

In the present specification, the “acyl or acyl group” may be either an aliphatic acyl group or an aromatic acyl group, or an aliphatic acyl group having an aromatic group as a substituent. Acyl groups may contain one or more heteroatoms. For example, as an acyl group, an alkylcarbonyl group (such as an acetyl group), an alkyloxycarbonyl group (such as an acetoxycarbonyl group), an arylcarbonyl group (such as a benzoyl group), an aryloxycarbonyl group (such as a phenyloxycarbonyl group), an aralkylcarbonyl group ( Benzylcarbonyl group), alkylthiocarbonyl group (such as methylthiocarbonyl group), alkylaminocarbonyl group (such as methylaminocarbonyl group), arylthiocarbonyl group (such as phenylthiocarbonyl group), or arylaminocarbonyl group (phenylaminocarbonyl group) And the like, but is not limited thereto. These acyl groups may have one or more arbitrary substituents. Examples of the substituent include, but are not limited to, an alkoxy group, a halogen atom, an amino group, a mono- or di-substituted amino group, a substituted silyl group, and an acyl group. When the acyl group has two or more substituents, they may be the same or different.

In the present specification, the “aryl or aryl group” may be either a monocyclic or condensed polycyclic aromatic hydrocarbon group, and a hetero atom (for example, an oxygen atom, a nitrogen atom, Or a sulfur atom or the like). In this case, it may be referred to as “heteroaryl” or “heteroaromatic”. Whether aryl is a single ring or a fused ring, it can be attached at all possible positions. In the present specification, an aryl group may have one or more arbitrary substituents on the ring. Examples of the substituent include, but are not limited to, an alkoxy group, a halogen atom, an amino group, a mono- or di-substituted amino group, a substituted silyl group, and acyl. When the aryl group has two or more substituents, they may be the same or different. The same applies to the aryl moiety of other substituents containing the aryl moiety (for example, an aryloxy group and an arylalkyl group).

In the present specification, the “alkoxy group” is a structure in which the alkyl group is bonded to an oxygen atom, and examples thereof include a saturated alkoxy group that is linear, branched, cyclic, or a combination thereof. For example, methoxy group, ethoxy group, n-propoxy group, isopropoxy group, cyclopropoxy group, n-butoxy group, isobutoxy group, s-butoxy group, t-butoxy group, cyclobutoxy group, cyclopropylmethoxy group, n- Pentyloxy group, cyclopentyloxy group, cyclopropylethyloxy group, cyclobutylmethyloxy group, n-hexyloxy group, cyclohexyloxy group, cyclopropylpropyloxy group, cyclobutylethyloxy group, cyclopentylmethyloxy group, etc. are preferable Take as an example.

As used herein, “amide” includes both RNR′CO— (where R = alkyl, alkylaminocarbonyl-) and RCONR′— (where R = alkyl, alkylcarbonylamino-).

As used herein, “ester” includes both ROCO— (in the case of R = alkyl, alkoxycarbonyl-) and RCOO— (in the case of R = alkyl, alkylcarbonyloxy-).

As used herein, the term “ring structure”, when formed by a combination of two substituents, means a heterocyclic or carbocyclic ring, such ring being saturated, unsaturated, or aromatic. be able to. Accordingly, it includes cycloalkyl, cycloalkenyl, aryl, and heteroaryl as defined above.

In the present specification, “phenolic OH group or phenolic hydroxyl group” means an OH group bonded to an arbitrary position on the benzene ring.

In the present specification, a specific substituent can form a ring structure with another substituent, and when such substituents are bonded to each other, those skilled in the art will recognize a specific substituent, for example, hydrogen. It can be understood that the bonds are formed. Therefore, when it is described that specific substituents together form a ring structure, those skilled in the art can understand that the ring structure can be formed by an ordinary chemical reaction and can be easily generated. Both such ring structures and their process of formation are within the purview of those skilled in the art. Moreover, the said heterocyclic structure may have arbitrary substituents on the ring.

2. The manufacturing method of the intermediate compound of a formula (I) This invention relates to the manufacturing method of the intermediate compound represented by the following formula | equation (I) in one aspect | mode.

The intermediate compound represented by the formula (I) has a pentacyclic skeleton in which two tetrahydroisoquinoline (THIQ) rings are connected. The XYZ moiety in the molecule is a side chain containing a long-chain alkyl necessary for proceeding with the enzyme reaction in Step A described later. Since saframycins such as saframycin Y3 and saframycin A have the pentacyclic skeleton as a common skeleton, the long chain alkyl moiety in XYZ is finally cleaved and removed. Saframycins are obtained. Therefore, a desired target product can be obtained by selecting the linker portions of X and Y together with the structure of the target product.

In the formula, X is —C (R ³ ) NH— or —C (R ³ ) O—; Y is —C (═O) R ⁴ —NH—, —C (═O) R ⁴ —. O—, —S (═O) ₂ R ⁴ —NH—, —S (═O) ₂ R ⁴ —O—, —P (═O) (OR ⁵ ) R ⁴ —NH—, or —P (= O) (OR ⁵ ) R ⁴ —O—. R ⁴ is an optionally substituted alkylene group. As described above, X and Y can be selected according to the desired structure of the target object. For example, as described later, when saframycin Y3 is the target product, X is —C (R ³ ) NH—, and Y is —C (═O) R ⁴ —NH—. Is preferred. Here, “—NHC (═O) O—” in the connecting portion of X and Y has a structure called carbamate or urethane. Thereby, the NH ₂ group at the end of saframycin Y3 can be obtained in one step. When saframycin A is the target product, X is preferably —C (R ³ ) NH— and Y is preferably —C (═O) R ⁴ —O—.

R ^{3 in} X is a hydrogen atom or an optionally substituted C ₁ -C ₅ alkyl group, preferably a hydrogen atom or a methyl group.

R ⁴ which is a linking group in Y is an optionally substituted alkylene group. A C ₂ -alkylene group is preferred, and 1-methylmethylene (—CH (CH ₃ ) —) is more preferred. Y is, ^R in the case of phosphoric acid esters or phosphoric acid amides, i.e. ^{^{-P (= O) (OR 5}} ) R 4 -NH- or ^{^{-P (= O) (OR 5}} ) For R 4 -O- ⁵ Is an optionally substituted C ₁ -C ₅ alkyl group, preferably methyl.

In the formula, Z is —C (═O) R ⁶ , —C (═O) OR ⁶ , —S (═O) ₂ R ⁶ , or —P (═O) (OR ⁵ ) R ⁶ . Where R ⁶ is a saturated or unsaturated hydrocarbon chain. As long as the enzymatic reaction in Step A described below proceeds, the number of carbon atoms of R ⁶ is not particularly limited, but is preferably a C ₅ -C ₃₀ alkyl group, and more preferably a C ₁₀ -C ₂₀ alkyl group. Moreover, it is preferable that it has an unsaturated double bond in the arbitrary positions of a hydrocarbon chain from a viewpoint of finally cut | disconnecting a side chain with a transition metal catalyst.

In the formula, R ¹ and R ² are each independently a C ₁ -C ₁₀ alkyl group which may be the same or different, preferably a methyl group. More preferably, each of R ¹ and R ² is preferably a methyl group. However, any substituent other than an alkyl group can be used depending on the final product of interest.

Where R ^a is a substituent on the tertiary amine in the pentacyclic skeleton. R ^a is preferably an optionally substituted alkyl group, and more preferably a methyl group. However, substituents other than alkyl groups can be used depending on the final product of interest.

The method for producing an intermediate compound represented by the formula (I) of the invention includes the following steps A to C. That is, in Step A, a precursor compound (II) having a pentacyclic skeleton in which two THIQ rings are linked by an enzymatic reaction is synthesized, and in Steps B and C, this is subjected to cyanation and reductive N-methylation. Thus, an intermediate compound represented by the formula (I) is obtained.

[Step A]: Enzyme-catalyzed reaction Step A comprises a compound represented by the following formula (a):

A compound represented by the following formula (b):

This is a step of obtaining a compound represented by the following formula (II) by reacting in the presence of a non-ribosome-dependent peptide synthase.

In these formulas, X, Y, Z, R ¹ and R ² are the same as those described for formula (I).

Step A comprises the steps of THIQ ring formation and pentacyclic skeleton formation from a compound represented by formula (a) which is a tyrosine derivative and a compound represented by formula (b) which is a non-natural aldehyde substrate. Reaction is performed to produce precursor compound (II). Here, the reaction is a biosynthetic reaction using an enzyme as a catalyst.

The enzyme used in Step A is a module constituting at least a part of a peptide synthase called non-ribosomal peptide synthetase (NRPS). NRPS is a huge multi-module enzyme complex with a molecular weight of several thousand kDa, and is known to synthesize short-chain physiologically active peptides without going through ribosomes. In general, a module, which is a repeating unit constituting NRPS, captures an activated amino acid as a thioester by recognizing and activating an amino acid serving as a substrate (A domain: adenylation domain), and forms a covalent bond. Basic composition of peptidyl carrier protein site (PCP site) (also referred to as T domain (thiolation domain)) that loads amino acid unit onto enzyme, and condensation domain (C domain condensation domain) that catalyzes the formation of peptide bond Element.

As the NRPS used in the present invention, SfmC which is a module of saframycin A biosynthetic enzyme is preferably used. This SfmC enzyme does not promote peptide formation, which is the original function, but imine formation between a long-chain fatty acid aldehyde and an amino acid (L-tyrosine derivative), followed by Pictet-Spenger (PS) -type cyclization It has been found by the inventors of the present invention that the enzyme has a unique property of forming a pentacyclic skeleton containing a THIQ ring by catalyzing (Non-patent Document 4: Koketsu, K. Watanabe, K .; Suda, H .; Oguri, H .; Oikawa, H., Nat. Chem. Biol., 2010, 6, 408).

As shown below, the SfmC enzyme is a PS domain that catalyzes the Pictet-Spengler reaction; an A domain that recognizes and activates L-tyrosine as a substrate; PCP that converts activated amino acids into thioesters and loads them onto the enzyme (Peptidyl carrier protein site); and a Red domain that reduces the thioester. The PS domain originally corresponds to the C domain (condensation domain) that catalyzes peptide bonds, but here it is referred to as a “PS domain” because it catalyzes a position- and stereoselective Pictet-Spangler reaction. ing.

The details of the reaction mechanism of pentacyclic skeleton formation by the SfmC enzyme are described in Non-Patent Document 4, and a method for producing the SfmC enzyme is also described. Therefore, those skilled in the art can understand that the SfmC enzyme is obtained and used by referring to the literature.

Further, examples of enzymes that can be used in addition to the SfmC enzyme include enzymes such as EtuA2 and SacC that have a catalytic function in the biosynthesis of ectenasaidin and safracin.

In order to perform the enzyme reaction of Step A of the present invention, it is preferable to add ATP, NADPH, Mg ²⁺ and Mn ²⁺ .

Specific examples of the non-natural aldehyde substrate represented by the formula (b) include the following compounds. However, it is not limited to this.

Wherein, ^{Z a} _is a C 6 _{-C 20} alkyl group, Me represents a methyl group.

[Step B]: Cyanation Reaction Step B is a step of adding a cyanide compound to the compound represented by the formula (II) obtained in Step A to convert an OH group on the pentacyclic skeleton to a cyano group (CN Group) to obtain a compound represented by the following formula (III).

In which X, Y, Z, R ¹ and R ² are the same as those described for formula (I).

As the cyan compound used in Step B, a compound containing cyanide ions can be used. Typically, an alkali metal cyanide such as potassium cyanide (KCN). In addition, a cyanating agent known in the art can be used.

[Step C]: N-alkylation reaction The compound represented by the formula (III) obtained in the above Step A is N-alkylated at the secondary amine moiety by adding an aldehyde and a reducing agent, In this step, a compound represented by (I) is obtained.

The aldehyde used in Step C is selected according to the type of alkyl group added by N-alkylation. For example, when adding a methyl group, formaldehyde is used. Moreover, as a reducing agent, the compound well-known in the said technical field used in a reductive amination reaction can be used. For example, a borohydride compound such as sodium triacetoxyborohydride or sodium cyanoborohydride can be used.

3. Method for Producing Saframycins Using Intermediate Compound In another aspect, the present invention also relates to a method for producing saframycins using an intermediate compound represented by the following formula (I). As specific examples, a method for producing saframycin Y3 and a derivative thereof (“saframycin Y3 compound”) and a method for producing saframycin A and a derivative thereof (“saframycin A compound”) will be described below.

(1) Manufacturing method of saframycin Y3 type | system | group compound The compound represented by the following formula | equation (IV) can be manufactured from the intermediate compound represented by the said formula (I).

(In the formula, X, R ¹ , R ² and R ^a have the same meaning as in formula (I), and A is a hydrogen atom or a protecting group.)

The compound of the formula (IV) has a structure corresponding to saframycin Y3, and the Z-site of the side chain is converted to A in the pentacyclic skeleton, and the end of Y is the amino group end or the hydrogen of the amino group A compound having an atom as a protecting group. As the protecting group for A, a protecting group known in the art such as a carbamate group such as Fmoc, Boc or Alloc (allyloxycarbonyl), an ester group, a sulfonate group or a silyl group can be used. . Even when A is a protecting group, the terminal can be changed to an amino group in the same manner as saframycin Y3 by subsequent deprotection.

From the viewpoint of performing such side chain conversion, in formula (I), X is —C (R ³ ) NH—, Y is —C (═O) R ⁴ —NH—, and Z is It is preferred to use a compound that is —C (═O) OR ⁶ . Other than these, R ¹ , R ² , and R ^a may be the same as those described above for formula (I).

The method for producing the saframycin Y3 compound includes the following steps D and E. In Step D, the acyl chain is removed from the Z site of the side chain and converted to an amino group or a protected amino group. In Step E, the phenol in the pentacyclic skeleton is converted to quinone.

[Step D]: Conversion reaction of side chain to amino group (removal of acyl chain)
In step D, the intermediate compound represented by formula (I) is converted to A which is an amino group or a protected amino group by removing the acyl chain from the Z site in the presence of a transition metal catalyst. In this step, a compound represented by formula (V) is obtained.

(In the formula, X, Y, A, R ¹ , R ² and R ^a have the same meaning as in formula (IV).)

It has been found for the first time in the present invention that the conversion from the Z site to A can be performed in one step by including “—NHC (═O) O—” in X and Y, that is, a carbamate structure or a urethane structure. It has been done. Conventionally, in the situation where there are many other functional groups, selective removal and amination of the terminal acyl chain has been difficult, and thus such a conversion reaction can be achieved for the first time by the present invention.

The transition metal catalyst used in Step D is preferably a palladium complex. Specific examples of the palladium complex include tetrakis (triphenylphosphine) palladium.

[Step E]: Oxidation reaction to quinone Step E oxidizes the compound represented by the formula (V) obtained in Step D in the presence of a transition metal catalyst to convert the phenol moiety in the pentacyclic skeleton to quinone. This is a step of obtaining a saframycin Y3 compound represented by the above formula (IV).

Here, the transition metal catalyst used is preferably a cobalt complex. Specific examples of the cobalt complex include sarcomin (N, N'-ethylenebis (salicylideneaminato) cobalt (II)), Fremy salt (potassium nitrosodisulfonate), and the like.

As described above, even when A in the formula (IV) is a protective group, by subsequent deprotection, A is returned to a hydrogen atom, and the terminal is changed to an amino group in the same manner as saframycin Y3. be able to. For such deprotection, a technique commonly used in this technical field can be used.

In the formula (IV), preferably, R ¹ , R ² and R ^a are all methyl groups; R ⁴ is —CH (CH ₃ ) — and A is a hydrogen atom. In this case, the compound of formula (IV) is saframycin Y3.

(2) Method for Producing Saframycin A-Based Compound From the intermediate compound represented by the above formula (I), a compound represented by the following formula (VI) can be produced.

(In the formula, X, R ¹ , R ² and R ^a have the same meaning as in formula (I).)

The compound of the formula (VI) has a structure corresponding to saframycin A, and is a compound having a carbonyl group terminal by converting the YZ site of the side chain into a pentacyclic skeleton. From the viewpoint of performing such side chain conversion, in formula (I), X is —C (R ³ ) NH—, Y is —C (═O) R ⁴ —O—, and Z is It is preferred to use a compound that is —C (═O) R ⁶ . Other than these, R ¹ , R ² , and R ^a may be the same as those described above for formula (I).

The method for producing the saframycin A compound includes the following steps F to H. In Step F, the acyl chain is removed from the YZ site of the side chain and converted to an OH group. In Step G, the phenol in the pentacyclic skeleton is converted to quinone. In Step H, the OH group at the end of the side chain is converted. Converted to a carbonyl group.

[Step F]: Conversion reaction of side chain to OH group (removal of acyl chain)
Step F is a compound represented by the following formula (VII) by hydrolyzing the compound represented by the formula (I) under basic conditions and converting it to the OH group terminal from which the YZ site acyl chain has been removed. A step of obtaining a compound to be produced. In the present invention, the conversion from the YZ site to the OH group end can be performed in one step by using “—OC (═O) —”, ie, an ester structure, as the linking part of Y and Z. It was discovered for the first time. Conventionally, in the situation where many other functional groups exist, selective removal of the terminal acyl chain and OH group formation have been difficult, and thus such a conversion reaction can be achieved for the first time by the present invention.

In order to achieve basic conditions in Step F, hydroxide ions (OH ⁻ ) are preferably added. Typically, an alkali metal hydroxide such as lithium hydroxide can be used.

[Step G]: Oxidation reaction to quinone Step G involves oxidation of the compound of formula (VII) obtained in Step F in the presence of a transition metal catalyst to convert the phenol moiety in the pentacyclic skeleton to quinone. And a step of obtaining a compound represented by the following formula (VIII). This process is the same reaction as in Step E above.

Here, the transition metal catalyst used is preferably a cobalt complex. Specific examples of the cobalt complex include sarcomin (N, N′-ethylenebis (salicylideneiminato) cobalt (II)), Fremy salt (potassium nitrosodisulfonate), and the like.

[Step H]: Carbonylation of terminal OH group Step H is a step of oxidizing the compound of formula (VIII) obtained in Step G using DMSO oxidation or a hypervalent iodine reagent to convert the terminal OH group to a carbonyl group. This is a step of obtaining a saframycin A-based compound represented by the formula (VI).

Hypervalent iodine reagent used in Step H, preferably a compound containing pentavalent iodine, typically 1,1,1-triacetoxy-1,1-dihydro-1, called Dess-Martin reagent Mention may be made of 2-benziodoxol-3 (1H) -one. DMSO oxidation can be performed using a known activator, and examples of the activator include oxalyl chloride. In addition to these, oxidation can be carried out using compounds and reagents well known in the art.

In the formula (I), preferably, R ¹ , R ² and ^Ra are all methyl groups; R ³ is a hydrogen atom. In this case, the final product, the compound of formula (VI), is saframycin A.

(3) Method for Producing Other Compounds As a modification of the method for producing the saframycin Y3-based compound described above, an intermediate compound represented by the above formula (I) is transformed into a side chain X- It is also possible to produce a compound in which the YZ moiety is converted to terminate the OH group. From the viewpoint of performing such side chain conversion, in formula (I), X is —C (R ³ ) O—, Y is —C (═O) R ⁴ —NH—, and Z is It is preferred to use a compound that is —C (═O) R ⁶ . Other than these, R ¹ , R ² , and R ^a may be the same as those described above for formula (I).

The manufacturing method of the OH group-terminated compound includes the following steps D2 and E2. In Step D2, the acyl chain is removed from the XYZ site of the side chain to convert it to an OH group, and in Step E2, the phenol site in the pentacyclic skeleton is converted to quinone.

[Step D2]: Side chain OH reaction (acyl chain removal)
In step D2, the intermediate compound represented by formula (I) is hydrolyzed under basic conditions to remove the acyl chain from the XYZ site of the side chain, and the XYZ site is This is a step of converting to a C (R ³ ) OH group.

In step D2, hydroxide ions (OH ⁻ ) are preferably added in order to achieve basic conditions. Typically, an alkali metal hydroxide such as lithium hydroxide can be used.

The conversion from the XYZ moiety to the primary hydroxyl group is carried out in one step by using “—O—C (═O) R ₃ ”, that is, an ester structure, as the linking part of X and Y. It has been found for the first time in the present invention. Conventionally, it has been difficult for the present invention to achieve such a conversion reaction for the first time because selective removal of an acyl chain and OH formation have been difficult in the presence of many other functional groups.

[Step E2]: Oxidation reaction to quinone Step E2 is a step of oxidizing the compound obtained in Step D2 in the presence of a transition metal catalyst to convert the phenol moiety in the pentacyclic skeleton to quinone. This process is the same reaction as in Step E above.

The reaction conditions such as the solvent and reaction temperature in each step of the production method of the present invention described above are described in detail as typical examples in the examples described later, but are not necessarily limited thereto. Those skilled in the art can appropriately select each based on general knowledge in organic synthesis.

When the non-natural aldehyde substrate represented by the formula (b) is selected from the following compounds:

The intermediate compound represented by formula (I) has the following structure.

(Wherein, ^{Z a} is a C ₆ -C ₂₀ alkyl group, Me represents a methyl group.)

Similarly, when the non-natural aldehyde substrate represented by formula (b) is selected from the above specific examples, the precursor compound represented by formula (II) has the following structure.

In another embodiment, when the non-natural aldehyde substrate represented by the formula (b) is selected from the following compounds:

Various intermediate compounds or products in the present invention have a plurality of asymmetric carbons, and there exist stereoisomers such as optical isomers or diastereoisomers, but pure forms of stereoisomers and stereoisomers are present. Any mixtures, racemates and the like are included within the scope of the present invention.

In addition, various intermediate compounds or products in the present invention may exist as hydrates or solvates, and any of these substances is included in the scope of the present invention. Although the kind of solvent which forms a solvate is not specifically limited, For example, solvents, such as ethanol, acetone, isopropanol, can be illustrated.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

[Reagents, equipment, etc.]
All reactions were performed under a nitrogen atmosphere unless otherwise noted. NMR spectra for JEOL JNM-ECA 500 (1H / 500MHz, 13C / 125MHz) spectrometer, Bruker VSP 500 (1H / 500MHz, 13C / 125MHz) spectrometer and Bruker AMX500 (1H / 500MHz, 13C / 125MHz) spectrometer Using. ^{In 1} H, ¹³ C-NMR, chloroform, acetonitrile and dimethyl sulfoxide were used as internal standards. ¹ H-NMR data are described as chemical shifts (hydrogen number, multiplicity, coupling constant). Multiplicity is described as s (singlet), d (doublet), t (triplet), q (quartet), quin (quintet), m (multiplet), br (broad) . For the ESI-mass spectrum, Bruker Daltonics micrOTOF-QII was used. Medium pressure liquid chromatography (MPLC) purification was performed with YAMAZEN YFLC-AI-580 and Biotage Isolera. Dess-Martin periodinane and sarcomin were obtained from Tokyo Chemical Industry Co., Ltd. ATP (disodium salt trihydrate) was purchased from Wako Pure Chemical Industries. EDC / HCl, HATU and HBTU were purchased from Watanabe Chemical Co., Ltd. Grubbs catalyst first generation and Pd (PPh ₃ ) ₄ were obtained from Sigma-Aldrich. NADPH and NADH were purchased from Oriental Yeast Co. Ltd. All reagents and commercially available solvents were used as received. The reaction was monitored by thin layer chromatography using Merck Millipore TLC silica gel F254 plates (0.25 mm). Flash column chromatography was performed using Kanto Silica Gel 60N.

1. Synthesis of Tyrosine Derivative Substrate The tyrosine derivative (Compound 1) was synthesized by using a non-patent document 5 (Tanifuji, R .; Oguri, H .; Koketsu, K .; Yoshinaga, Y .; Minami, A .; Oikawa, H. Tetrahedron Lett. 2016 , 57, 623.).

2. Synthesis of aldehyde substrates

(1) Synthesis of Aldehyde Substrate 1 (Compound 11) Aldehyde substrate 1 (Compound 11) was synthesized from Compound 6 according to the following reaction scheme.

[Synthesis of Compound 4]

A suspension of 2 (281 mg, 0.557 mmol) and 10% Pd / C (178 mg, 0.167 mmol, 30 mol%) in THF (10 mL) was stirred under low hydrogen pressure (0.60 MPa) at room temperature for 1.5 hours. The catalyst was removed from the mixture by Celite filtration and concentrated under reduced pressure to give 3, which was used without further purification.
CH _₂ Cl ₂ (10mL) solution of crude 7 and _{Et 3 N (0.621mL, 4.46mmol,} 8.0 eq) to a suspension of, CH _₂ Cl ₂ (10mL) solution of Alloc-OSu (222mg, 1.11mmol, 2.0 equivalents) was added and stirred at room temperature for 13.5 hours. Water (20 mL) and 1 M HCl aqueous solution 5.0 mL were added to the obtained mixture, and the mixture was extracted with CH ₂ Cl ₂ (50 mL × 3). The organic extract was washed with saturated brine, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography (hexane / AcOEt) to give 4 (224 mg, 0.492 mmol, 88% yield over 2 steps) as a white solid.
¹ H NMR (500 MHz, CDCl ₃ , δ): 1.06 (9H, s), 1.36 (3H, d, J = 6.9 Hz), 3.36-3.46 (2H, m), 3.74 (2H, t, J = 5.2 Hz), 4.11-4.24 (1H, m), 4.49-4.62 (2H, m), 5.16-5.33 (3H, m), 5.85-5.95 (1H, m), 6.30 (1H, br), 7.36--7.47 (6H, m), 7.61-7.66 (4H, m) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 19.00, 19.33, 26.97, 41.73, 50.70, 62.64, 66.03, 118.11, 128.00, 130.06, 132.65 , 133.26, 133.30, 135.64, 155.89, 172.16 .; HRMS (ESI, m / z): [M + Na] ⁺ calcd.for C ₂₅ H ₃₄ N ₂ O ₄ SiNa, 477.2186; found, 477.2173.

[Synthesis of Compound 5]

To the reflux solution of the first generation of Grubbs catalyst (222 mg, 0.267 mmol, 5.0 mol%), 4 (2.45 g, 5.40 mmol) and 1-undecene (11.1 mL, 54.0 mmol, 10 eq) CH ₂ Cl ₂ (20 mL ) Added to solution. The mixture was heated to reflux for 17.5 hours and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / AcOEt) to obtain 5 (2.45 g, 4.22 mmol, 78% yield) as a pale yellow oil.
¹ H NMR (500 MHz, CDCl _3, δ): 0.88 (3H, t, J = 6.9 Hz), 1.06 (9H, s), 1.20-1.31 (12H, m), 1.32-1.41 (4H, m), 2.03 (2H, q, J = 7.5 Hz), 3.36-3.47 (2H, m), 3.74 (2H, t, J = 5.2 Hz), 4.13-4.25 (1H, m), 4.41-4.57 (2H, m) , 5.14 (1H, br), 5.49-5.60 (1H, m), 5.70-5.79 (1H, m), 6.33 (1H, s), 7.35-7.48 (6H, m), 7.60-7.67 (4H, m) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 14.28, 18.95, 19.33, 22.83, 26.97, 29.05, 29.33, 29.48, 29.63, 29.70, 32.04, 32.41, 41.70, 50.68, 62.64, 66.25, 124.04, 127.99 , 130.05, 133.25, 133.30, 135.42, 135.64, 136.75, 156.08, 172.22 .; HRMS (ESI, m / z): [M + Na] ⁺ calcd. For C ₃₄ H ₅₂ N ₂ O ₄ SiNa, 603.3588; found, 603.3573.

[Synthesis of Compound 6]

To a solution of 5 (280 mg, 0.482 mmol) in THF (15.0 mL, 0.032 M) was added 1 M TBAF in THF (0.627 mL, 0.627 mmol, 1.3 eq). The mixture was stirred at room temperature for 45 minutes. It was then quenched with saturated aqueous NH ₄ Cl (10 mL) and stirred at room temperature for 5 minutes. The mixture was concentrated under reduced pressure, 20 mL of water was added, and the mixture was extracted with CH ₂ Cl ₂ (50 mL × 3). The organic extract was washed with saturated brine, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography (CHCl ₃ / MeOH) to give 6 (127 mg, 0.372 mmol, 77% yield) as a white solid.
¹ H NMR (500 MHz, CDCl ₃ , δ): 0.87 (3H, t, J = 7.5 Hz), 1.19-1.41 (17H, m), 2.02 (2H, q, J = 6.9 Hz), 3.29-3.50 ( 2H, m), 3.69 (1H, br), 4.13-4.30 (1H, m), 4.41-4.54 (2H, m), 5.45-5.57 (2H, m), 5.70-5.80 (1H, m), 6.88 ( ¹³ C NMR (125 MHz, CDCl ₃ , δ): 14.26, 18.75, 22.81, 29.04, 29.33, 29.46, 29.61, 29.69, 29,83, 32.03, 32.41, 42.25, 50.80, 61.82, 66.37 , 123.87, 136.90, 156.45, 173.62 .; HRMS (ESI, m / z): [M + Na] ⁺ calcd.for C ₁₈ H ₃₄ N ₂ O ₄ Na, 365.2411; found, 365.2394.

[Synthesis of Compound 7]

To a solution of 6 (30.9 mg, 0.0902 mmol) in CH ₂ Cl ₂ (10 mL, 0.0090 M) was added Dess-Martin periodinane (134 mg, 0.316 mmol, 3.5 eq). The mixture was stirred at 0 ° C. for 1 hour. The resulting mixture was quenched with 10% Na ₂ S ₂ O ₃ saturated aqueous NaHCO ₃ (5.0 mL) and stirred at 0 ° C. for 15 min. The mixture was extracted with CH ₂ Cl ₂ (30 mL × 3) and the organic extract was washed with brine, dried over Na ₂ SO ₄ and concentrated in vacuo. The crude residue was purified by silica gel column chromatography (CH ₂ Cl ₂ / AcOEt) to give compound 7 (22.0 mg, 0.0646 mmol, 72% yield) as a white solid.
¹ H NMR (500 MHz, CDCl ₃ , δ): 0.87 (3H, t, J = 6.9 Hz), 1.16-1.43 (18H, m), 19.98-2.11 (2H, m), 4.19 (2H, t, J = 5.2 Hz), 4.25-4.36 (1H, m), 4.44-4.55 (2H, m), 5.33 (1H, m), 5.46-5.58 (2H, m), 5.69-5.81 (2H, m), 6.95 ( 1H, br), 9.64 (1H, s) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 14.25, 18.56, 22.80, 27.69, 29.01, 29.31, 29.45, 29.60, 29.67, 29.82, 32.02, 32.39, 50.27, 50.50, 66.37, 123.48, 123.89, 135.56, 136.89, 156.26, 173.01, 196.36 .; HRMS (ESI, m / z): [M + K] ⁺ calcd.for C ₁₈ H ₃₂ N ₂ O ₄ K, 379.1994 ; found, 379.1984.

(2) Synthesis of Aldehyde Substrate 2 (Compound 11) Aldehyde substrate 11 (Compound 4) was synthesized from Compound 8 according to the following reaction scheme.

[Synthesis of Compound 9]

An EtOH solution (10 mL, 0.85 M) of ethyl lactate 8 (962 μL, 8.47 mmol) and allylamine (1.90 mL, 25.4 mmol, 3.0 equivalents) was stirred at 120 ° C. for 35 hours under microwave irradiation. After the ethanol was distilled off under reduced pressure, the liquid residue was roughly purified by passing it through silica gel using ethyl acetate. After concentration under reduced pressure, the crude residue was purified by silica gel column chromatography (CHCl ₃ / AcOEt) to give 9 (1.01 g, 7.83 mmol, 92%) as a colorless oil.
¹ H NMR (500 MHz, CDCl ₃ , δ): 1.36 (3H, d, J = 6.9 Hz), 3.81-3.83 (2H, m), 4.17 (1H, m), 4.58 (1H, s), 5.08- 5.16 (2H, m), 5.74-5.81 (1H, m), 7.05 (1H, br) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 21.16, 41.41, 68.28, 116.43, 133.75, 175.39 .; HRMS (ESI, m / z): [M + Na] ⁺ calcd. For C ₆ H ₁₁ NO ₂ Na, 152.0682; found, 152.0688.

[Synthesis of Compound 10]

MNBA (2.52 g, 7.32 mmol, 1.2 eq), myristic acid (2.03 g, 7.32 mmol, 1.2 eq), DMAP (149 mg, 1.22 mmol, 20 mol%) and triethylamine (2.13 mL, 15.2 mmol, 2.5 eq) A CH ₂ Cl ₂ solution (15.3 mL, 0.30 M) of 9 (788 mg, 6.10 mmol) was added to the CH ₂ Cl ₂ solution (5.0 mL), and the mixture was stirred at room temperature for 13 hours. After cooling to 0 ° C., saturated aqueous NaHCO ₃ (15 mL) was added to terminate the reaction. The resulting mixture was extracted with CH ₂ Cl ₂ (50 mL × 3). The organic extract was washed with saturated brine, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography (CHCl ₃ / AcOEt, hexane / AcOEt) to obtain 10 (1.41 g, 4.15 mmol, 68%) as a white solid.
¹ H NMR (500 MHz, CDCl ₃ , δ): 0.86 (3H, t, J = 6.9 Hz), 1.24-1.32 (20H, m), 1.46 (3H, d, J = 6.9 Hz), 1.60-1.66 ( 2H, m), 2.36 (2H, t, J = 7.4 Hz), 3.89 (2H, m,), 5.13-5.24 (3H, m), 5.78-5.86 (1H, m), 6.21 (1H, br). ; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 14.21, 18.06, 22.78, 24.99, 29.18, 29.34, 29.45, 29.54, 29.69, 29.74, 29.76, 32.02, 34.43, 41.60, 70.49, 116.67, 133.87, 170.45, 172.39 .; HRMS (ESI, m / z): [M + Na] ⁺ calcd. For C ₂₀ H ₃₇ NO ₃ Na, 362.2665; found, 362.2652.

[Synthesis of Compound 11]

Ozone was bubbled through a solution of 10 (209 mg, 0.616 mmol) in CHCl ₃ (140 mL, 4.4 mM) at −30 ° C. until the solution turned blue-violet. Excess ozone was removed from the solution by bubbling oxygen and nitrogen. After repeating this process once more, Me ₂ S (1.37 mL, 18.5 mmol, 30 eq) was added and stirred at −30 ° C. for 10 minutes and at room temperature for 40 minutes. After concentration under reduced pressure, the crude residue was purified by silica gel column chromatography (CHCl ₃ / AcOEt) to obtain 11 (168 mg, 0.493 mmol, 80%) as a white solid.
[α] _D ²⁹ -7.1 ° (c 1.0, CHCl ₃ ); ¹ H NMR (500 MHz, CDCl ₃ , δ): 0.85 (3H, t, J = 6.9 Hz), 1.17-1.29 (20H, m), 1.42-1.46 (3H, m), 1.61-1.67 (2H, m), 2.35-2.40 (2H, m), 4.20 (2H, d, J = 4.6 Hz), 5.24 (1H, q, J = 6.9 Hz) , 6.89 (1H, s), 9.65 (1H, s) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 14.19, 17.95, 22.76, 24.93, 29.17, 29.32, 29.42, 29.52, 29.66, 29.72, 29.74 , 32.00, 34.37, 49.99, 70.23, 171.06, 172.44, 195.83 .; HRMS (ESI, m / z): [M + Na] ⁺ calcd. For C ₁₉ H ₃₅ NO ₄ Na, 364.2458; found, 364.2447.

3. Synthesis of Saframycin Y3 Saframycin Y3 (Compound 16 ′) was synthesized from the tyrosine derivative (Compound 1) obtained in Example 1 and the aldehyde substrate 1 (Compound 7) obtained in Example 2. The holoenzyme SfmC used in the reaction is described in Non-Patent Document 4 (Koketsu, K .; Watanabe, K .; Suda, H .; Oguri, H .; Oikawa, H. Nat. Chem. Biol. 2010, 6, 408). ).

[Synthesis of Compound 13]

50 mM KH ₂ PO ₄ -KOH buffer containing 13.7 μM Holified SfmC, 1.0 mM Compound 1, 400 μM Compound 7 (added as DMSO solution), 20 mM MgCl ₂ , 20 μM MnCl ₂ , 2.0 mM ATP 2.0 mM NADPH was added to a 20 mL solution of (pH 7.5) to initiate the enzyme reaction and incubated at 30 ° C. for 2 hours. The reaction mixture was treated with 100 mM KCN aqueous solution (750 μL) and incubated at 30 ° C. for 5 minutes. The resulting mixture was diluted with MeCN (60 mL) and then centrifuged at 7000 × g for 5 minutes to remove SfmC. This was filtered to give a solution containing compound 12, which was used without further purification.

To a suspension of crude compound 12 and 37% aqueous HCHO (39.8 μL, 14.7 mg, 50 eq) in MeCN (60 mL), water (20 mL) was added 2-picoline borane (42.8 mg, 0.40 mmol, 50 Equivalent) was added and stirred at room temperature for 30 minutes. The resulting mixture was concentrated under reduced pressure. Water (20 mL) was added to the residue and extracted with Et ₂ O (30 mL × 3). The organic extract was washed with saturated brine and dried over Na ₂ SO ₄ . After filtration and concentration in vacuo, the crude product was passed through Sep-Pak® C18 and eluted with MeCN. After concentration, the residue was purified by HPLC using Inertsilsustain C18 (GL Sciences, φ10 × 250 mm, 5 μm) under the following conditions: increasing the ratio of MeCN to water from 30% to 75% with a linear gradient for 35 minutes It was. At this time, the flow rate was 5.0 mL / min, the column temperature was room temperature, and the wavelength of the UV detector was 280 nm. Fractions containing the desired product were concentrated under reduced pressure to give compound 13 (1.37 mg, 1.84 μmol, 23% yield over 3 steps). The resulting compound 13 was analyzed by a series of NMR measurements including ¹ H-NMR, ¹³ C-NMR, H-COSY, HSQC, HMBC and ROESY.
¹ H NMR (500 MHz, CDCl ₃ , δ): 0.87 (3H, t, J = 6.9 Hz), 0.93 (3H, d, J = 6.9 Hz), 1.20-1.39 (17H, m), 2.02 (2H, q, J = 6.9 Hz), 2.09-2.19 (1H, m), 2.21 (3H, s), 2.27 (3H, s), 2.30 (3H, s), 2.56 (1H, d, J = 18.3 Hz), 2.80 (1H, dd, J = 15.5, 2.3 Hz), 3.05 (1H, dd, J = 18.0, 8.6 Hz), 3.20-3.33 (2H, m), 3.36 (1H, m), 3.45 (1H, m) , 3.67-3.75 (4H, m), 3.79 (3H, s), 4.03 (1H, d, J = 2.3 Hz), 4.00-4.05 (1H, m), 4.14 (1H, br), 4.34-4.46 (2H , m), 5.16 (1H, br), 5.35 (1H, br), 5.43-5.54 (1H, m), 5.66-5.76 (1H, m), 5.85 (1H, s), 6,00 (1H, s ), 6.42 (1H, s), 6.49 (1H, s) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 14.29, 15.83, 15.94, 19.14, 22.83, 25.51, 25.62, 29.08, 29.34, 29.48, 29.63, 29.70, 32.04, 32.11, 32.42, 41.53, 41.92, 50.18, 55.46, 56.63, 56.75, 57.00, 59.96, 60.92, 60.95, 64.61, 65.90, 111.80, 117.13, 117.59, 118.22, 121.14, 121.28, 124.18, 121.129 131.02, 131.93, 136.45, 143.20, 143.66, 145.09, 146.99, 155.51, 171.68 .; HRMS (ESI, m / z): [M + H] ⁺ calcd. For C ₄₃ H ₆₃ N ₄ O ₇ , 747.4692; found, 747.4698.

[Synthesis of Compound 14]

To a solution of Pd (PPh ₃ ) ₄ (9.91 mg, 8.58 μmol, 50 mol%) and phenylsilane (5.29 μL, 42.9 μmol, 2.5 eq) in CH ₂ Cl ₂ (0.50 mL), compound 13 (12.8 mg, 17.1 μmol) of CH ₂ Cl ₂ (1.0 mL) was added. The mixture was stirred at 30 ° C. for 20 minutes, SiliaMetS Thiourea (registered trademark, 85.1 mg, 103 μmol, 6.0 eq. SILI CYCLE) was added, and the mixture was stirred at room temperature for 50 minutes. After the resulting mixture was filtered and concentrated under reduced pressure, the residue was passed through Sep-Pak® C18, eluting with MeCN to give a solution containing compound 14, which was used without further purification.
To a solution of compound 14 in _{_{CH 2 Cl 2 (1.0mL),}} Fmoc-OSu (6.37 mg, 18.9 μmol, 1.1 eq) was added CH _₂ Cl ₂ (1.0mL) solution of. The mixture was stirred at room temperature for 6.5 hours, then passed through Sep-Pak® C18 and eluted with MeCN. After concentration, the residue was purified by HPLC using Inertsustain C18 (GL Sciences, φ10 × 250 mm, 5 μm) under the following conditions: Increase the ratio of MeCN to water from 30% to 75% with a linear gradient for 30 minutes. It was. At this time, the flow rate was 5.0 mL / min, the column temperature was room temperature, and the wavelength of the UV detector was 280 nm. Fractions containing the desired product were concentrated under reduced pressure to give compound 15 (10.4 mg, 13.7 μmol, 80% yield over 2 steps). The resulting compound 15 was analyzed by a series of NMR measurements including ¹ H-NMR, ¹³ C-NMR, H-COSY, HSQC, HMBC and ROESY.
¹ H NMR (500 MHz, CDCl ₃ , δ): 0.96 (3H, d, J = 6.9 Hz), 2.11 (3H, s), 2.15-2.20 (1H, m), 2.25 (3H, s), 2.31 ( 3H, s), 2.55 (1H, d, J = 18.3 Hz), 2.80 (1H, dd, J = 15.5, 2.3 Hz), 3.05 (1H, dd, J = 18.3, 8.0 Hz), 3.26--3.32 ( 2H, m), 3.36 (1H, d, J = 8.0 Hz), 3.46 (1H, t, J = 6.6 Hz), 3.66 (3H, s), 3.73-3.82 (4H, m), 4.03 (1H, d , J = 2.3 Hz), 4.09-4.18 (3H, m), 4.21--4.34 (2H, m), 5.27 (1H, d, J = 6.3 Hz), 5.36 (1H, s), 5.86 (1H, s ), 5.98 (1H, s), 6.38 (1H, s), 6.47 (1H, s), 7.27-7.34 (2H, m), 7.35-7.44 (2H, m), 7.47-7.60 (2H, m), 7.75 (2H, d, J = 7.4 Hz) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 15.72, 15.93, 19.12, 25.67, 32.11, 41.52, 41.91, 47.21, 50.29, 55.47, 56.64, 56.77, 56.92, 59.94, 60.81, 60.93, 66.97, 117.19, 117.63, 118.18, 120.12, 121.08, 121.24, 125.17, 125.24, 127.18, 127.83, 129.24, 130.99, 131.90, 141.37, 141.42, 143.21, 143.67, 143.89, 144.07, 145.1 147.06, 155.50, 171.70 .; HRMS (ESI, m / z): [M + H] ⁺ calcd. For C ₄₄ H ₄₈ N ₅ O ₇ , 758.3548; found, 758.3557.

[Synthesis of saframycin Y3]

A solution of compound 15 (10.3 mg, 13.6 μmol) and salcomine (4.42 mg, 13.6 μmol, 1 eq) in MeCN (10 mL), ethyl acetate (2 mL) was bubbled with oxygen gas for 5 min at room temperature. After stirring for 2.5 hours in an oxygen atmosphere, oxygen gas was blown again for 3 minutes. After stirring in an oxygen atmosphere for 30 minutes, SiliaMetS Thiourea (registered trademark, 89.9 mg, 108 μmol, 8.0 eq. SILI CYCLE) was added and stirred for 15 minutes. After the resulting mixture was filtered and concentrated under reduced pressure, the residue was passed through Sep-Pak (registered trademark) Florisil and eluted with AcOEt. After concentration, the residue was purified by HPLC using Inertsil Diol (GL Sciences, φ10 × 250 mm, 5 μm) under the following conditions: The ratio of AcOEt acetate to hexane was 30% to 60% in a straight line for 30 minutes. Increased with gradient. At this time, the flow rate was 5.0 mL / min, the column temperature was room temperature, and the wavelength of the UV detector was 280 nm. Fractions containing the desired product were concentrated under reduced pressure to give compound 16 (6.87 mg, 8.74 μmol, 64% yield). The resulting compound 16 was analyzed by a series of NMR measurements including ¹ H-NMR, ¹³ C-NMR, H-COSY, HSQC, HMBC and ROESY. ¹ H NMR (500 MHz, CDCl ₃ , δ): 1.19 (3H, d, J = 6.9 Hz), 1.81 (3H, s), 1.90 (3H, s), 2.30-2.37 (4H, m), 2.75- 2.85 (2H, m), 3.09 (1H, dt, J = 11.3, 3.0 Hz), 3.29 (1H, m), 3.42 (1H, d, J = 7.7 Hz), 3.66-3.72 (2H, m), 3.85 -3.95 (4H, m), 4.00 (3H, s), 4.06-4.14 (4H, m), 4.22-4.25 (1H, m), 4.91 (1H, d, J = 7.4 Hz), 5.90 (1H, br ), 7.29-7.43 (4H, m), 7.60 (1H, d, J = 7.4 Hz), 7.71 (1H, d, J = 7.4 Hz), 7.76 (2H, d, J = 7.4 Hz) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 8.7, 8.8, 17.7, 21.4, 25.2, 40.1, 41.7, 47.1, 50.9, 54.1, 54.3, 54.7, 57.1, 58.2, 61.1, 61.2, 67.4, 117.0, 120.1, 120.1 , 125.3, 125.4, 127.2, 127.3, 127.85, 127.90, 135.67, 135.74, 141.4, 141.6, 143.6, 144.3, 155.8, 156.2, 172.6, 181.3, 182.7, 185.5, 187.0 .; HRMS (ESI, m / z): [ M + H] + calcd.for C ₄₄ H ₄₄ N ₅ O ₉ , 786.3134; found, 786.3110.

Compound 16 (1.69 mg, 2.15 μmol) was stirred in 5% piperidine / DMF (0.50 mL) at room temperature for 40 minutes. The resulting mixture was passed through Sep-Pak® C18 and eluted with MeCN. This solution was analyzed by LC-MS to confirm the formation of saframycin Y3 (compound 16 ′).

4). Synthesis of Saframycin A Saframycin A (Compound 21) was synthesized from the tyrosine derivative (Compound 1) obtained in Example 1 and the aldehyde substrate 2 (Compound 11) obtained in Example 2. The holoenzyme SfmC used in the reaction is described in Non-Patent Document 4 (Koketsu, K .; Watanabe, K .; Suda, H .; Oguri, H .; Oikawa, H. Nat. Chem. Biol. 2010, 6, 408). ).

[Synthesis of Compound 18]

14.2 μM Holated SfmC, 1.0 mM Compound 1, 400 μM Compound 11 (added as DMSO solution), 20 mM MgCl ₂ , 20 μM MnCl ₂ , 50 mM KH ₂ PO ₄ containing 2.0 mM ATP -To a 120 mL solution of KOH buffer (pH 7.5), 2.0 mM NADPH was added to initiate the enzyme reaction and incubated at 30 ° C for 2 hours. The reaction mixture was treated with 100 mM KCN aqueous solution (750 μL) and incubated at 30 ° C. for 5 minutes. The resulting mixture was diluted with MeCN (360 mL) and then centrifuged at 7000 × g for 5 minutes to remove SfmC. This was filtered to give a solution containing compound 17, which was used without further purification.
To a suspension of crude compound 17 and 37% aqueous HCHO (195 μL, 72.0 mg, 2.40 mmol, 50 equiv) in MeCN (360 mL), water (120 mL) was added 2-picoline borane (103 mg, 0.96 mmol, 20 equivalents) was added and stirred at room temperature for 30 minutes. The resulting mixture was concentrated under reduced pressure. The residue was extracted with AcOEt (80 mL × 3). The organic extract was washed with saturated brine and dried over Na ₂ SO ₄ . After filtration and concentration in vacuo, the crude product was passed through Sep-Pak® C18 and eluted with MeCN. After concentration, the residue was purified by HPLC using Inertsustain C18 (GL Sciences, φ10 × 250 mm, 5 μm) under the following conditions: Increase the ratio of MeCN to water from 30% to 75% with a linear gradient for 30 minutes. It was. At this time, the flow rate was 5.0 mL / min, the column temperature was room temperature, and the wavelength of the UV detector was 280 nm. Fractions containing the desired product were concentrated under reduced pressure to give compound 18 (4.58 mg, 6.13 μmol, 13% yield over 3 steps). The resulting compound 18 was analyzed by a series of NMR measurements including ¹ H-NMR, ¹³ C-NMR, H-COSY, HSQC, HMBC and ROESY. ¹ H NMR (500 MHz, CDCl ₃ , δ): 0.88 (3H, t, J = 6.9 Hz), 1.16-1.31 (23H, m), 1.38-1.43 (2H, m), 1.99-2.06 (3H, m ), 2.23-2.28 (9H, m), 2.67 (1H, d, J = 18.3 Hz), 2.78 (1H, dd, J = 15.5, 1.7 Hz), 2.94 (1H, q, J = 8.8 Hz), 3.20 -3.28 (1H, m), 3.30-3.36 (1H, m), 3.37-3.44 (1H, m), 3.64-3.70 (1H, m), 3.74-3.82 (6H, m), 4.06 (2H, d, J = 2.6 Hz), 4.15 (1H, s), 4.32 (1H, q, J = 6.9 Hz), 5.58-5.66 (1H, m), 5.71 (1H, s), 5.88 (1H, s), 6.42 ( 1H, s), 6.47 (1H, s) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 14.26, 15.84, 15.96, 17.67, 22.83, 24.87, 25.17, 29.27, 29.34, 29.50, 29.66, 29.75, 29.80, 32.06, 32.22, 34.14, 40.14, 41.91, 55.46, 56.40, 56.57, 57.57, 59.84, 60.90, 60.95, 70.09, 116.66, 117.62, 118.45, 121.24, 121.35, 128.79, 129.14, 131.21, 132.143, 142.75, 145.11, 146.58, 170.14, 171.47 .; HRMS (ESI, m / z): [M + H] ⁺ calcd. For C ₄₃ H ₆₃ N ₄ O ₇ , 747.4691; found, 747.4706.

[Synthesis of Compound 19]

An aqueous solution (1.0 mL) of LiOH.H ₂ O (2.05 mg, 48.8 μmol, 3.0 eq) was added to compound 18 (12.2 mg, 16.3 μmol) in t-BuOH (3.0 mL) and stirred at room temperature for 7 hours. Further, an aqueous solution (0.50 mL) of LiOH.H ₂ O (1.37 mg, 32.6 μmol, 2.0 equivalents) was added and stirred for 1 hour. 200 mM NaH ₂ PO ₄ .NaOH aqueous solution (2.0 mL) was added, and the mixture was stirred at room temperature for 5 minutes to complete the reaction. After concentration under reduced pressure, water (10 mL) was added to the resulting residue, and the mixture was extracted with AcOEt (30 mL × 3). The organic extract was washed with saturated brine and dried over Na ₂ SO ₄ . After filtration and concentration in vacuo, the crude product was passed through Sep-Pak® C18 and eluted with MeCN. After concentration, the residue was purified by HPLC using Inertsustain C18 (GL Sciences, φ10 × 250 mm, 5 μm) under the following conditions: Increase the ratio of MeCN to water from 30% to 75% with a linear gradient for 30 minutes. It was. At this time, the flow rate was 5.0 mL / min, the column temperature was room temperature, and the wavelength of the UV detector was 280 nm. Fractions containing the desired product were concentrated under reduced pressure to give compound 19 (7.93 mg, 14.8 μmol, 91% yield). The resulting compound 19 was analyzed by a series of NMR measurements including ¹ H-NMR, ¹³ C-NMR, H-COSY, HSQC, HMBC and ROESY. ¹ H NMR (500 MHz, CDCl ₃ , δ): 0.98 (3H, d, J = 6.9 Hz), 2.13 (1H, m), 2.22 (3H, s), 2.26 (3H, s), 2.30 (3H, s), 2.58 (1H, d, J = 17.8 Hz), 2.81 (1H, dd, J = 15.5, 2.3 Hz), 3.02 (1H, q, J = 8.8 Hz), 3.30-3.37 (2H, m), 3.42--3.49 (1H, m), 3.55 (1H, q, J = 6.7 Hz), 3.60-3.67 (1H, m), 3.73 (3H, s), 3.78 (3H, s), 4.04 (1H, d , J = 2.3 Hz), 4.11 (1H, d, J = 2.3 Hz), 4.16 (1H, br), 5.68-5.87 (2H, m), 5.98 (1H, br), 6.42 (1H, s), 6.50 (1H, s) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 15.81, 15.85, 20.85, 25.49, 32.16, 40.61, 41.90, 55.45, 56.60, 56.66, 56.92, 59.85, 60.90, 60.94, 68.25, 117.37, 117.71, 118.24, 121.04, 121.23, 128.86, 129.28, 131.48, 131.84, 142.80, 143.79, 145.24, 146.93, 174.40 .; HRMS (ESI, m / z): [M + H] ⁺ calcd. For C ₂₉ H ₃₇ N ₄ O ₆ , 537.2708; found, 537.2711.

[Synthesis of Saframycin A]

A solution of compound 19 (7.61 mg, 10.2 μmol) and salcomine (4.38 mg, 13.5 μmol, 1.0 equiv) in MeCN (5.0 mL) was bubbled with oxygen gas for 5 minutes at room temperature. After stirring for 1.5 hours under an oxygen atmosphere, SiliaMetS Thiourea (registered trademark, 89.1 mg, 108 μmol, 8.0 equivalents) was added and stirred for 15 minutes at room temperature. After the resulting mixture was filtered and concentrated under reduced pressure, the residue was passed through Sep-Pak® C18, eluting with MeCN to give a solution containing compound 20, which was used without further purification.
To a solution of oxalyl chloride (11.6 μL, 17.1 mg, 135 μmol, 10 equivalents) in CH ₂ Cl ₂ (4.0 mL) was added dimethyl sulfoxide (14.4 μL, 15.8 mg, 202 μmol, 15 equivalents) dropwise at −78 ° C. Stir for 5 minutes. A CH ₂ Cl ₂ solution (1.0 mL) of a crude product of Compound 19 (7.34 mg) (1.0 mL) was slowly added dropwise to this mixture at −78 ° C., and the mixture was stirred for 15 minutes. Triethylamine (56.4 μL, 40.9 mg, 404 μmol, 30 equivalents) was added to the obtained mixture at −78 ° C., and the mixture was stirred for 10 minutes. The reaction solution was heated to 0 ° C. over 50 minutes, then passed through Sep-Pak (registered trademark) Florisil and eluted with AcOEt. After concentration, the residue was purified by HPLC using Inertsil Diol (GL Sciences, φ10 × 250 mm, 5 μm) under the following conditions: The ratio of AcOEt acetate to hexane was 30% to 60% in a straight line for 30 minutes. Increased with gradient. At this time, the flow rate was 5.0 mL / min, the column temperature was room temperature, and the wavelength of the UV detector was 280 nm. Fractions containing the desired product were concentrated under reduced pressure to give compound 19 (4.97 mg, 8.83 μmol, 66% yield over 2 steps). The resulting compound 19 was analyzed by a series of NMR measurements including ¹ H-NMR, ¹³ C-NMR, H-COSY, HSQC, HMBC and ROESY. ¹ H NMR (500 MHz, CDCl ₃ , δ): 1.24-1.32 (2H, m), 1.91 (3H, s), 1.98 (3H, s), 2.22-2.26 (4H, m), 2.31 (3H, s ), 2.79-2.90 (2H, m), 3.13 (1H, dt, J = 11.5, 2.9 Hz), 3.26 (1H, dt, J = 14.3, 4.0 Hz), 3.43 (1H, d, J = 7.4 Hz) , 3.72 (1H, m), 3.97 (1H, s), 3.99 (1H, d, J = 2.3 Hz), 4.02 (6H, m), 4.06 (1H, m), 6.67 (1H, br) .; ¹³ C NMR (125 MHz, CDCl ₃ , δ): 8.88, 21.63, 24.43, 25.18, 40.75, 41.73, 54.07, 54.31, 54.60, 56.40, 58.37, 61.11, 61.27, 116.78, 128.46, 129.37, 135.68, 135.71, 141.36, 141.66, 155.77, 156.05, 160.31, 180.96, 182.60, 185.44, 186.75, 196.85 .; HRMS (ESI, m / z): [M + H] ⁺ calcd.for C ₂₉ H ₃₁ N ₄ O ₈ , 563.2136; found, 563.2152.

Claims

A method for producing a compound represented by the following formula (I):

Wherein X is —C (R 3 ) NH— or —C (R 3 ) O—; Y is —C (═O) R 4 —NH—, —C (═O) R 4 —O—, —S (═O) 2 R 4 —NH—, —S (═O) 2 R 4 —O—, —P (═O) (OR 5 ) R 4 —NH—, or —P (═O) (OR 5 ) R 4 —O—; Z is —C (═O) R 6 , —C (═O) OR 6 , —S (═O) 2 R 6 , or —P (═O) (OR 5 ) R 6 ; R 1 and R 2 are each independently a C 1 -C 10 alkyl group, which may be the same or different; R 3 is a hydrogen atom or An optionally substituted C 1 -C 5 alkyl group; R 4 is an optionally substituted alkylene group; R 5 is an optionally substituted C 1 -C 5 alkyl group; ; R 6 is a saturated also A hydrocarbon chain of unsaturated; R a is optionally substituted alkyl group).
[Step A] a compound represented by the following formula (a):

(Wherein R 1 and R 2 have the same meaning as in formula (I).)
A compound represented by the following formula (b):

(In the formula, X, Y and Z have the same meaning as in formula (I).)
A step of obtaining a compound represented by the following formula (II) by reacting in the presence of a non-ribosome-dependent peptide synthase

(Wherein X, Y, Z, R 1 and R 2 have the same meaning as in formula (I));
[Step B] A step of adding a cyanide ion to the compound represented by the formula (II) to obtain a compound represented by the following formula (III)

(Wherein X, Y, Z, R 1 and R 2 have the same meaning as in formula (I)); and [Step C] an aldehyde and a reducing agent for the compound represented by formula (III) A process for producing a compound represented by the formula (I) by N-alkylating a secondary amine moiety by adding
The production method according to claim 1, wherein the non-ribosome-dependent peptide synthase is saframycin A biosynthetic enzyme SfmC.
The manufacturing method of Claim 1 or 2 including adding ATP, NADPH, Mg <2+> , and Mn < 2+ > in the said step A. FIG.
The production method according to any one of claims 1 to 3, wherein R 1 and R 2 are both methyl groups.
The production method according to any one of claims 1 to 4, wherein the compound represented by the formula (b) is selected from the group consisting of the following compounds.

(Wherein, Z a is a C 6 -C 20 alkyl group, Me represents a methyl group.)
From the intermediate compound represented by the following formula (I)

Wherein X is —C (R 3 ) NH—; Y is —C (═O) R 4 —NH—; Z is —C (═O) OR 6 ; R 1 and R 2 are each independently a C 1 -C 10 alkyl group, which may be the same or different; R 3 is a hydrogen atom or an optionally substituted C 1 -C 5 alkyl group R 4 is an optionally substituted alkylene group; R 6 is a saturated or unsaturated hydrocarbon chain; R a is an optionally substituted alkyl group.)
A method for producing a compound represented by the following formula (IV):

(Wherein X, Y, R 1 , R 2 and R a have the same meaning as in formula (I), and A is a hydrogen atom or a protecting group);
[Step D] A step of obtaining a compound represented by the following formula (V) by converting the Z site of the compound represented by the formula (I) into A in the presence of a transition metal catalyst.

(In the formula, X, Y, A, R 1 , R 2 and R a have the same meaning as in formula (IV).)
[Step E] The production comprising the step of obtaining the compound represented by the formula (IV) by oxidizing the compound represented by the formula (V) in the presence of a transition metal catalyst and converting phenol to quinone. Method.
The production method according to claim 6, wherein R 1 , R 2 and R a are all methyl groups; R 3 is a hydrogen atom; and R 4 is —CH (CH 3 ) —.
The production method according to claim 6 or 7, wherein the transition metal catalyst in Step D is a palladium complex.
The production method according to any one of claims 6 to 8, wherein the transition metal catalyst in Step E is a cobalt complex.
From the intermediate compound represented by the following formula (I)

Wherein X is —C (R 3 ) NH—; Y is —C (═O) R 4 —O—; Z is —C (═O) R 6 ; R 1 and R 2 are each independently a C 1 -C 10 alkyl group, which may be the same or different; R 3 is a hydrogen atom or an optionally substituted C 1 -C 5 alkyl group R 4 is a 1-methylmethylene group; R 6 is a saturated or unsaturated hydrocarbon chain; R a is an optionally substituted alkyl group.)
A method for producing a compound represented by the following formula (VI):

(Wherein X, R 1 , R 2 and R a have the same meaning as in formula (I));
[Step F] A compound represented by the following formula (VII) is obtained by hydrolyzing the compound represented by the above formula (I) under basic conditions and converting the end of the Y site into an OH group. Process

(In the formula, X, R 1 , R 2 and R a have the same meaning as in formula (I).)
[Step G] A step of obtaining a compound represented by the following formula (VIII) by oxidizing the compound represented by the formula (VII) in the presence of a transition metal catalyst and converting phenol to quinone.

(Wherein X, R 1 , R 2 and R a have the same meaning as in formula (I)); and [Step H] The compound represented by (VIII) is subjected to DMSO oxidation or hypervalence. This manufacturing method including the process of obtaining the compound represented by said Formula (VI) by converting the terminal OH group into a carbonyl group by oxidation using an iodine reagent.
The production method according to claim 10, wherein R 1 , R 2 and Ra are all methyl groups.
The production method according to claim 10 or 11, wherein the transition metal catalyst in Step G is a cobalt complex.
The compound represented by the following formula (I).

Wherein X is —C (R 3 ) NH— or —C (R 3 ) O—; Y is —C (═O) R 4 —NH—, —C (═O) R 4 —O—, —S (═O) 2 R 4 —NH—, —S (═O) 2 R 4 —O—, —P (═O) (OR 5 ) R 4 —NH—, or —P (═O) (OR 5 ) R 4 —O—; Z is —C (═O) R 6 , —C (═O) OR 6 , —S (═O) 2 R 6 , or —P (═O) (OR 5 ) R 6 ; R 1 and R 2 are each independently a C 1 -C 10 alkyl group, which may be the same or different; R 3 is a hydrogen atom or a substituent An optionally substituted C 1 -C 5 alkyl group; R 4 is an optionally substituted alkylene group; R 5 is an optionally substituted C 1 -C 5 alkyl group; R 6 is a saturated or A hydrocarbon chain, saturated; R a is optionally substituted alkyl group).
14. A compound according to claim 13, selected from the group consisting of:

(Wherein, Z a is a C 6 -C 20 alkyl group, Me represents a methyl group.)
A compound represented by the following formula (II).

Wherein X is —C (R 3 ) NH— or —C (R 3 ) O—; Y is —C (═O) R 4 —NH—, —C (═O) R 4 —O—, —S (═O) 2 R 4 —NH—, —S (═O) 2 R 4 —O—, —P (═O) (OR 5 ) R 4 —NH—, or —P (═O) (OR 5 ) R 4 —O—; Z is —C (═O) R 6 , —C (═O) OR 6 , —S (═O) 2 R 6 , or —P (═O) (OR 5 ) R 6 ; R 1 and R 2 are each independently a C 1 -C 10 alkyl group, which may be the same or different; R 3 is a hydrogen atom or a substituent An optionally substituted C 1 -C 5 alkyl group; R 4 is an optionally substituted alkylene group; R 5 is an optionally substituted C 1 -C 5 alkyl group; R 6 is a saturated or A hydrocarbon chain, saturated; R a is optionally substituted alkyl group).
16. A compound according to claim 15 selected from the group consisting of:

(Wherein, Z a is a C 6 -C 20 alkyl group, Me represents a methyl group.)
An aldehyde compound represented by the following formula (b).

Wherein X is —C (R 3 ) NH— or —C (R 3 ) O—; Y is —C (═O) R 4 —NH—, —C (═O) R 4 —O—, —S (═O) 2 R 4 —NH—, —S (═O) 2 R 4 —O—, —P (═O) (OR 5 ) R 4 —NH—, or —P (═O) (OR 5 ) R 4 —O—; Z is —C (═O) R 6 , —C (═O) OR 6 , —S (═O) 2 R 6 , or —P (═O) (OR 5 ) R 6 ; R 3 is a hydrogen atom or an optionally substituted C 1 -C 5 alkyl group; R 4 is an optionally substituted alkylene group R 5 is an optionally substituted C 1 -C 5 alkyl group; R 6 is a saturated or unsaturated hydrocarbon chain.)
The aldehyde compound according to claim 17, which is selected from the group consisting of the following compounds.

(Wherein, Z a is a C 6 -C 20 alkyl group, Me represents a methyl group.)