JP2007238506A - New compound and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new compound having stronger ubiquitylation inhibitory activity than that of panepophenanthrin and a method for efficiently producing the new compound or panepophenanthrin. <P>SOLUTION: The new panepophenanthrin derivative compound is claimed. The method for producing the same comprises a first reaction process in which a carbonyl compound is reacted with a nitroso compound in the presence of an asymmetric catalyst and a second reaction process in which the product generated by the first reaction process is stereoselectively reacted to produce the derivative of the new compound. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to novel compounds. Specifically, the present invention relates to a novel compound having ubiquitination inhibitory activity and a method for producing the same.

  The ubiquitin-proteasome system that regulates proteolysis plays an important role in the mechanism of eukaryotic growth control. Abnormal proteolytic pathways via the ubiquitin-proteasome system are often associated with cancer, inflammation, and neurological diseases. It is considered as one of the disease factors (see Non-Patent Document 1).

  Ubiquitin is composed of highly conserved 76 amino acid residues, and is derived from ubiquitin activating enzyme (hereinafter referred to as E1 enzyme), ubiquitin-binding enzyme (hereinafter referred to as E2 enzyme), and ubiquitin ligase (hereinafter referred to as E3 enzyme). Is added to the target protein to form a polyubiquitin chain, and the target protein is finally degraded by the proteasome. The E1 enzyme activates a cysteine residue present at the C-terminus of monomeric ubiquitin to form a thioester, and then transfers it to a reactive cysteine residue of the E2 enzyme. The E2 enzyme cooperates with the E3 enzyme. Acts to bind ubiquitin to the amino group of a lysine residue on the target protein.

  Therefore, the E1 enzyme is an important enzyme in the early stage of the ubiquitination pathway, and a drug that inhibits E1 activity can be a therapeutic agent for various diseases that are considered to be caused by abnormalities in the ubiquitin-proteasome system. So far, panepophenanthrin has been reported as an E1 enzyme inhibitor (see Non-Patent Document 2).

Further, total synthesis of this compound has already been studied in various places (see Non-Patent Documents 3 to 6).
Drug Discovery Today 2001, 6, 244 J. et al. Nat. Prod. 2002, 65, 1491 Lei, X. Johnson, R .; P. Porco, J .; A. , Jr. , Angew. Chem. , Int. Ed. 2003, 42, 3913. Moses, J. et al. E. Commeiras, L .; Baldwin, J .; E. Addington, R .; M.M. Org. Lett. 2003, 5, 2987. (A) Mehta, G .; Ramesh, S .; S. Tetrahedron Lett. 2004, 45, 1985. (B) Mehta, G .; Islam, K .; Tetrahedron Lett. 2004, 45, 7683.

  However, panepofenanthrin disclosed in Patent Document 2 has poor cell membrane permeability, and in order to use it as a pharmaceutical, a large amount of panepofenanthrin must be added.

  In addition, when synthesizing panepophenanthrin, the method disclosed in Non-Patent Document 3 discloses a method in which asymmetric epoxidation is performed in order to obtain an optically active substance. It requires 6 equivalents of a chiral auxiliary and is not suitable for mass synthesis. Furthermore, the synthesis method disclosed in Non-Patent Document 4 is a racemic synthesis, and a required optically active substance is not synthesized. Furthermore, the methods disclosed in Non-Patent Documents 5 and 6 perform optical resolution using an enzyme, and half of the product is an unnecessary enantiomer. This enantiomer is not environmentally friendly and expensive to manufacture.

  In view of the above problems, an object of the present invention is to provide a novel compound having ubiquitination inhibitory activity stronger than that of panepofenanthrin. Another object of the present invention is to provide an efficient method for producing this novel compound and panepophenanthrin.

  The present inventors can efficiently synthesize a novel substance having a higher ubiquitination-inhibiting activity than that of panepophenanthrin from the obtained derivative by using the raw material of panepophenanthrin and an asymmetric catalyst. As a result, the present invention has been completed. In addition, when panepophenanthrin was produced via a method for synthesizing this derivative, it was found that it could be produced more efficiently than the conventional synthesis method, and the present invention was completed. . Specifically, the following are provided.

［式中、Ｒ_１は水素原子、官能基を有していてもよい炭素数１から３０のアルキル基、置換基を有していてもよい炭素数６から１２のアリール基、置換基を有していてもよい炭素数６から１２のヘテロアリール基からなる群から選ばれるいずれか１種の基である。］ (1) A novel compound having a structure represented by the general formula (1).

[Wherein R ₁ has a hydrogen atom, an alkyl group having 1 to 30 carbon atoms which may have a functional group, an aryl group having 6 to 12 carbon atoms which may have a substituent, or a substituent. And any one group selected from the group consisting of heteroaryl groups having 6 to 12 carbon atoms. ]

Panepophenanthrin has a structure represented by the following structural formula (2). According to the invention of (1), by making R of the novel compound an alkyl group, an aryl group, or a heteroaryl group, it becomes possible to impart higher ubiquitination inhibitory activity than panepophenanthrin.

  (2) A ubiquitination inhibitor comprising the novel compound according to (1) as an active ingredient.

  According to the invention of (2), by containing the above-mentioned novel compound as an active ingredient, a ubiquitination inhibitor having a more immediate effect than a ubiquitination inhibitor containing panepofenanthrin as an active ingredient is produced. It becomes possible. Moreover, it becomes possible to obtain a ubiquitination inhibitor having the same activity even in an amount smaller than that of panepofenanthrin.

  (3) A pharmaceutical comprising the ubiquitination inhibitor according to (2) as an active ingredient.

  According to the invention of (3), by including the novel compound as an active ingredient, it becomes possible to produce a more effective drug than a drug containing panepofenanthrin as an active ingredient. In addition, it is possible to obtain a medicine that exhibits the same effect even in an amount smaller than that of panepofenanthrin. As a result, it is possible to provide a pharmaceutical with less side effects.

  (4) A method for producing a novel compound having a structure represented by the general formula (1), wherein a carbonyl compound and a nitroso compound are reacted in the presence of an asymmetric catalyst, and the first reaction. A method for producing a novel compound having a structure represented by the general formula (1), comprising: a second reaction step in which a product produced by the step is stereoselectively reacted to obtain a derivative of the novel compound.

  It is important to control the stereoselectivity of the product when producing panepophenanthrin and the novel compounds described in (1) to (3). Therefore, according to the invention of (4), it is possible to obtain an optically active compound by reacting the first reaction step in the presence of an asymmetric catalyst. As a result, the second reaction step can be stereoselectively advanced. This makes it possible to obtain panepophenanthrin and novel compounds with high yield.

  (5) The method for producing a novel compound according to (4), wherein the asymmetric catalyst is proline.

  Proline is a catalyst that imparts high stereoselectivity. Therefore, according to the invention of (5), since the asymmetric catalyst in the first reaction step was proline, the first reaction step was reacted in the presence of proline, thereby having high optical activity. It becomes possible to obtain a compound. In addition, since proline is inexpensive, it is possible to industrially synthesize panepophenanthrin and new compounds on an industrial scale. Since proline is a stable substance, it can be synthesized safely and there is little concern about environmental pollution due to organic waste liquid.

  (6) The second reaction step includes a reduction step in which the aminooxy group in the product obtained in the first reaction step is reductively cleaved to a hydroxyl group, and an epoxidation step in which the hydroxyl group is epoxidized. The method for producing a novel compound according to (4) or (5), which comprises a step of introducing a side chain into the epoxy compound obtained by this epoxidation step.

  According to the invention of (6), the second reaction step is a step including a reduction step, an epoxidation step and a side chain introduction step, so that higher stereoselectivity can be imparted.

  According to the present invention, an optically active compound can be obtained by reacting the first reaction step in the presence of an asymmetric catalyst. As a result, the second reaction step can be stereoselectively advanced. This makes it possible to obtain panepophenanthrin and novel compounds with high yield.

  Hereinafter, embodiments of the present invention will be described.

［式中、Ｒ_１は水素原子、官能基を有していてもよい炭素数１から３０のアルキル基、置換基を有していてもよい炭素数６から１２のアリール基、置換基を有していてもよい炭素数６から１２のヘテロアリール基からなる群から選ばれるいずれか１種の基である。］ [New substance]
The novel compound according to the present invention is represented by the general formula (1).

[Wherein R ₁ has a hydrogen atom, an alkyl group having 1 to 30 carbon atoms which may have a functional group, an aryl group having 6 to 12 carbon atoms which may have a substituent, or a substituent. And any one group selected from the group consisting of heteroaryl groups having 6 to 12 carbon atoms. ]

  Here, the “alkyl group having 1 to 30 carbon atoms which may have a functional group” means a methyl group, an ethyl group, an n-propyl group, an isopropyl group, a cyclopropyl group, an n-butyl group, an isobutyl group, Examples thereof include linear, branched or cyclic alkyl groups such as t-butyl group, n-pentyl group, isopentyl group, neopentyl group, t-pentyl group, isoamyl group and n-hexyl group. Examples of the alkyl group which may have a functional group include a silyl group substituted with an alkyl group or an aryl group, an alkoxy group, an amino group, an amide group, and a hydroxyl group.

  The “aryl group having 6 to 12 carbon atoms which may have a substituent” refers to a tolyl group, a 1-naphthyl group, a 2-naphthyl group, or an aryl group such as a methyl group, an ethyl group, Examples thereof include a linear or branched alkyl group such as an n-propyl group and an isopropyl group, or a group substituted with an alkyl group which may have a functional group as described above.

  The “optionally substituted heteroaryl group having 6 to 12 carbon atoms” is a heteroaryl group, which is selected from an oxygen atom, a nitrogen atom, and a sulfur atom. A monocyclic or bicyclic aromatic group containing a heteroatom. The heteroaryl group refers to a group that may be substituted with one or more halogen atoms. Specific examples include furan, thiophene, imidazole, and indole. Among them, R is preferably a hydrogen atom or an alkyl group having 4 to 8 carbon atoms.

[Production method]
The novel compound represented by the general formula (1) is produced from a derivative produced by the production method shown below. From this derivative, it is possible to produce not only a novel compound but also panepophenanthrin.

<Synthesis of derivatives>
The derivative comprises a first reaction step in which a carbonyl compound and a nitroso compound are reacted in the presence of an asymmetric catalyst, and a product produced by the first reaction step is reacted in a stereoselective manner to produce the new compound. It is produced through a second reaction step to obtain a derivative.

［式中、Ｒ_２，Ｒ_３はそれぞれ独立して炭素数１から２３のアルキル基であり、Ｒ_２とＲ_３が一緒になって環を形成していてもよい。］ In the first reaction step, the “carbonyl compound” that is the starting material is a compound that reacts with the asymmetric catalyst. The carbonyl compound is preferably a cyclic compound having 8 to 30 carbon atoms having a carbonyl group and two or more oxy groups or thio groups. Furthermore, the carbonyl group and the oxy group or thio group in the cyclic compound are preferably arranged in separate alicyclic rings. Specifically, it is preferably represented by the following structural formula (3) or (4).

[Wherein R ₂ and R ₃ are each independently an alkyl group having 1 to 23 carbon atoms, and R ₂ and R ₃ may be combined to form a ring. ]

The “alkyl group” for R ₂ and R ₃ may be linear or branched. Specifically, as described above, methyl group, ethyl group, n-propyl group, isopropyl group, cyclopropyl group, n-butyl group, isobutyl group, t-butyl group, n-pentyl group, isopentyl group, neopentyl group. , T-pentyl group, isoamyl group, n-hexyl group and the like, straight chain, branched chain or cyclic alkyl group.

The “nitroso compound”, which is another starting material in the first reaction step, refers to a compound represented by the following general formula (5). Here, R ₄ represents an aryl group, a heterocyclic ring, an alkyl group, an alkenyl group, or an alkynyl group. All of these may have a substituent.

  Examples of the aryl group include an optionally substituted phenyl group and naphthyl group. Of these, a phenyl group is preferred. The substituent is preferably a linear or branched alkyl group having 1 to 15 carbon atoms.

  Examples of the heterocyclic ring include piperidine, furan, thiophene, pyrrole, pyrazole, imidazole, triazole, oxazole, isoxazole, thiazole, isothiazole, dioxolane, pyridine, pyrimidine, pyrazine, triazine, dioxane, dithiane, morpholine, azepine , Oxepin, thiepin and the like.

  As described above, the aryl group and the heterocyclic group may have a substituent. Examples of such a substituent include an alkyl group, an alkenyl group, a nitro group, a halogen atom (for example, a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom).

  The alkyl group preferably has 1 to 20 carbon atoms, and particularly preferably has 1 to 5 carbon atoms. Specifically, for example, methyl group, ethyl group, propyl group, isopropyl group, n-butyl group, sec-butyl group, t-butyl group, pentyl group, hexyl group, heptyl group, n-octyl group, 2- Ethylhexyl, t-octyl, nonyl, decyl, dodecyl, tridecyl, tetradecyl, pentadecyl, n-hexadecyl, 2-hexyldecyl, heptadecyl, octadecyl, nonadecyl, icosyl, etc. Can be mentioned.

  The alkenyl group preferably has 2 to 20 carbon atoms, more preferably about 2 to 5 carbon atoms. Examples of the alkenyl group include propenyl groups such as vinyl group and allyl group, butyryl group, pentenyl group, hexenyl group, heptenyl group, octenyl group, nonenyl group, decenyl group, undecenyl group, dodecenyl group, tridecenyl group, tetradecenyl group, Examples include a pentadecenyl group, a hexadecenyl group, a heptadecenyl group, an octadecenyl group, a nonadecenyl group, and an icocenyl group. The alkenyl group may further have a substituent, and examples of such a substituent include the aryl group and the heterocyclic group described above.

Further, the alkynyl group represented by R ₄ is preferably one having 2 to 20 carbon atoms, particularly preferably 2 to 5 carbon atoms.

  Among such nitroso compounds, nitrosobenzene is preferably used.

In this first reaction step, proline or a proline derivative having a structure represented by the following general formula (6) can be used as an asymmetric catalyst. However, in view of stereoselectivity and reactivity, proline Is preferably used.

  In the above formula, A represents a hydrogen atom, an alkoxy group, an aryloxy group, an acyloxy group or a silyloxy group which may have a substituent. Here, as an alkoxy group, a C1-C20 thing, for example, a methoxy group, an ethoxy group, a propoxy group, a butoxy group, a pentyloxy group etc. are mentioned. In addition, examples of the aryloxy group include a phenyloxy group and a naphthyloxy group. Examples of the acyloxy group include an acetoxy group and a benzoyloxy group. Examples of the substituent for the silyloxy group include an alkyl group, an aryl group, and an alkenyl group.

  As reaction conditions in the first reaction step, first, a carbonyl compound as a starting material and a nitroso compound are dissolved in an organic solvent to prepare a solution. Here, the proline or proline derivative which is an asymmetric catalyst is preferably used in an amount of 0.01 to 1 equivalent, more preferably 0.05 equivalent and 0.3 equivalent, based on the nitroso compound.

Examples of the organic solvent used here include dimethylformamide, dimethyl sulfoxide, nitromethane (CH ₃ NO ₂ ), N-methyl-pyrrolidinone, acetonitrile (CH ₃ CN), chloroform (CHCl ₃ ), dichloromethane (CH ₂ Cl ₂ ), and the like. The polar solvent is preferably, but not limited to.

  The resulting reaction solution of ketone and proline or proline derivative is preferably cooled to −50 ° C. to 25 ° C., preferably −10 ° C. to 10 ° C., and this temperature is preferably maintained during the next reaction. In this reaction, when L-proline is used, α-aminooxyketone is the (R) isomer as the main product, and when D-proline is used, the α-aminooxyketone is the (S) isomer as the main product. Become.

  The substance produced | generated by this 1st reaction process has high stereoselectivity.

  Next, the product obtained in the first reaction step is stereoselectively reacted in the second reaction step to synthesize the novel compound or panepophenanthrin derivative according to the present invention.

  The second reaction step includes a reduction step for reducing the product obtained in the first reaction step, a protective group deprotection, dehydration, hydroxyl group protection, an epoxidation step, a double bond introduction and a side chain. And an introduction step.

  First, the “reduction step” refers to a step of reductively cleaving the aminooxy group in the product obtained in the first reaction step to form a hydroxyl group. At this time, it is preferable to use palladium / carbon, platinum / carbon, palladium hydroxide or the like as a reducing agent in a hydrogen atmosphere. Alternatively, copper sulfate may be used.

  The reaction temperature is preferably from -10 ° C to 100 ° C, and more preferably from 0 ° C to 30 ° C. Furthermore, the reaction time is preferably 5 minutes to 120 hours, more preferably 1 hour to 10 hours.

  As the solvent, it is preferable to use methanol, tetrahydrofuran, acetone, ethanol or the like.

  Next, the “epoxidation step” refers to a step of epoxidizing the alkene obtained by deprotecting and dehydrating the protecting group of the product obtained by the reduction step. In this case, benzyltrimethylammonium hydroxide, 1,8-diazabicyclo [5.4.0] undec-7-ene, 1,4-diazabicyclo [2.2.2] octane, etc. are used as usable catalysts. It is preferable.

  The reaction temperature is preferably -40 ° C to 100 ° C, more preferably -10 ° C to 30 ° C. Furthermore, the reaction time is preferably 10 minutes to 120 hours, more preferably 5 minutes to 3 hours.

  As the solvent, it is preferable to use methanol, tetrahydrofuran, acetone, ethanol or the like.

  Next, the “side chain introduction step” refers to a step of introducing a side chain into the epoxy compound obtained by the epoxidation step. The introduction of a side chain leads the epoxy compound to a silyl enol ether and converts it to a cyclohexenone derivative by an oxidation reaction. A halogen atom is introduced into the α-position of the ketone, and the reaction is carried out by Still reaction or Suzuki reaction.

  Here, the side chain to be introduced is, for example, 1-alkenyl such as 1-ethynyl, 1-propenyl, 1-branyl and the like.

  The reaction temperature is preferably 0 ° C. to 200 ° C., more preferably 30 ° C. to 150 ° C. Furthermore, the reaction time is preferably 1 minute to 24 hours, and more preferably 1 minute to 1 hour.

  As the solvent, it is preferable to use toluene, tetrahydrofuran or the like.

  The derivative thus obtained makes it possible to efficiently obtain panepophenanthrin and the novel compound according to the present invention.

[Production method of ubiquitination inhibitor]
The novel compounds according to the present invention can be used as ubiquitination inhibitors. For example, the ubiquitination inhibitory effect can be further enhanced by containing it in a pharmaceutical that does not have a very strong ubiquitination inhibitory activity.

  As a specific method for producing a ubiquitination inhibitor, a novel compound or panepofenensulin is produced by the above-described method, and a predetermined amount is added to a pharmaceutical product that does not have a strong ubiquitination inhibitory activity. A preferable addition amount is preferably from 0.01 mg to 10 mg per 100 mg of drug, and more preferably from 0.1 mg to 1 mg per 100 mg of drug.

[Pharmaceutical manufacturing method]
As for pharmaceuticals, the ubiquitination inhibitor according to the present invention can be administered as it is or diluted with water or the like. Alternatively, it can be formulated with a known pharmaceutical carrier.

  The dose of the ubiquitination inhibitor in the medicament depends on the patient's age, condition, and weight, and the mode of application. In general, the daily dose of active ingredient is preferably about 0.5 mg / kg to 50 mg / kg body weight for oral application and about 0.1 mg / kg to 10 mg / kg body weight for parenteral application. .

  Examples of pharmaceutical dosage forms include powders, granules, fine granules, dry syrups, tablets, capsules, ointments, injections, and eye drops. These pharmaceuticals are suitable excipients, disintegrants, binders, lubricants, diluents, buffering agents, tonicity agents, preservatives, wetting agents, depending on the dosage form, depending on the method used in pharmacology. It can be produced by mixing or diluting / dissolving appropriately with pharmaceutical additives such as agents, emulsifiers, dispersants, stabilizers, solubilizers, etc., and preparing according to conventional methods.

  For example, in the preparation of pharmaceuticals, disintegrating agents such as magnesium stearate, calcium stearate, sodium stearyl fumarate and polyethylene glycol waxes and lubricants may be added to the ubiquitination inhibitor according to the present invention. Furthermore, lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose derivatives, gelatin and the like may be mixed.

  When prepared into capsules, they are preferably prepared in vegetable oil, fat or other soft gelatin capsules with capsules containing a mixture of suitable excipients. Hard gelatin capsules may also contain granules of the active compound.

  When preparing a syrup or suspension, it is preferable to add sugar or sugar alcohol and ethanol, water, glycerol, propylene glycol, polyethylene glycol, and mixtures thereof. It may further contain colorants, flavoring agents, preservatives, saccharin and carboxymethylcellulose or other thickening agents. The solution may also be prepared in the form of a dry powder that dissolves in a suitable solvent prior to use.

  When preparing as a solution for parenteral administration, it is prepared as a solution in which a ubiquitination inhibitor is dissolved in a pharmaceutically acceptable solvent. These solutions also contain a stabilizing component and / or a buffering component. A solution for parenteral administration may be prepared as a dry preparation that dissolves in an appropriate solvent before use.

全ての反応は、アルゴン雰囲気下で行なわれた。また、生成物は、薄層クロマトグラフィーにより確認した。また、生成物の同定は、偏光計（日本分光社製）、ＦＴ−ＩＲスペクトル（株式会社堀場製作所製）、ＮＭＲスペクトル（ブルカー・バイオスピン株式会社、日本電子株式会社製）を使用した。 [Synthesis of novel compounds and panepophenanthrin]
The novel compound and panepophenanthrin according to the present invention are synthesized from a derivative represented by (10) in the following synthesis scheme (A). Hereinafter, the reaction at each stage will be described. The compound numbers shown below are all based on this synthesis scheme (A).

All reactions were performed under an argon atmosphere. The product was confirmed by thin layer chromatography. Moreover, the product was identified using a polarimeter (manufactured by JASCO Corporation), an FT-IR spectrum (manufactured by Horiba, Ltd.), and an NMR spectrum (manufactured by Bruker BioSpin Corporation, manufactured by JEOL Ltd.).

<Synthesis of (R) -7-anilinoxy-1,4-dioxa-spiro [4.5] decan-8-one>
(R) -7-anilinoxy-1,4-dioxa-spiro [4.5] decan-8-one represented by the structural formula (4) in the above synthesis scheme (A) was synthesized. The synthesis scheme is as follows.

  First, the compound 1,4-dioxa-spiro [4.5] decan-8-one (275 mg, 2.4 mmol) represented by the structural formula (3) in the above synthesis scheme and D-proline (23 mg, 0.2 mg). Dimethylformamide solution (3 ml) in which nitrosobenzene (214 mg, 2.0 mmol) is dissolved is gradually added to a dimethylformamide solution (9.0 ml) with 20 mmol) over 12 hours at 0 ° C. . Stir for 30 minutes after addition. The reaction was then terminated by adding a pH 7 phosphate buffer. Then, ethyl acetate was added to extract the product, and the product was further washed with brine. The washed product was dehydrated by adding anhydrous sodium sulfate. These were filtered and distilled under reduced pressure to purify. Purification was performed by chromatography (mobile phase: ethyl acetate: hexane = 1: 10 to 1: 4 mixture). The yield of the obtained product was 93%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 1.88-2.04 (2H, m), 2.16 (1H, t, J = 12.8 Hz), 2.36-2.46 (2H, m), 2.62 (1H, dt, J = 14.0, 6.8 Hz), 4.38-4.21 (4H, m), 4.60 (1H, dd, J = 12.9, 6.5 Hz) 6.87 (2H, d, J = 7.7 Hz), 6.90 (1H, t, J = 7.2 Hz), 7.20 (2H, t, J = 7.2 Hz);
¹³ C-NMR (CDCl ₃ ): δ 34.9, 36.0, 39.7, 64.8, 64.9, 82.7, 107.6, 114.5, 122.2, 128.9, 148 0.0, 208.6;
IR (KBr): 2960, 2888, 1728, 1602, 1494, 1305, 1122, 1052 cm ⁻¹ ;
[Α] _D ¹⁸ +78.7 (c = 1.2, CHCl ₃ ),> 99% ee;
HRMS (FAB): calculated value [C ₁₄ H ₁₇ NO ₄ ]: 263.1158, measured value: 263.1172.

The enantiomeric excess was calculated using high performance liquid chromatography. The measurement conditions at this time are as follows.
Column: OD-H column (hexane, 2-propanol 10: 1 mixed solution)
Flow rate: 0.5 mL / min
major enantiomer tr = 26.5 min
minor enantiomer tr = 29.1 min

<Synthesis of 1,4-dioxa-spiro [4.5] decane-7,8-diol>
1,4-dioxa-spiro [4.5] decane-7,8-diol represented by the structural formula (6) in the above synthesis scheme (A) was synthesized. The synthesis scheme is as follows.

Tri (sec-butyl) boron hydride as a reducing agent was added as a reducing agent to a tetrahydrofuran solution (77 ml) in which the α-aminoxyketone (3.0 g, 12 mmol) represented by the structural formula (4) synthesized by the above method was dissolved. Potassium (trademark: K-selectride) (23.1 ml, 23.1 mmol) was added at −78 ° C. Subsequently, the temperature of the solution was raised to -50 degreeC and made to react for 1.5 hours. Next, MeBO ₃ (10.8 g, 0.0701 mmol) and water (23 ml) were added to the reaction solution, and the mixture was stirred at room temperature for 2 hours. Thereafter, each of the phases separated into two phases was extracted, the organic phase was washed with brine, and dehydrated by adding anhydrous sodium sulfate. 4.6 g of the obtained product was directly used in the next reaction without purification in order to substitute with alcohol.

  Methanol (38 ml) was added to the above product, and 304 mg of palladium / carbon catalyst (10 wt%) was further added, followed by stirring at room temperature for 3 hours under a hydrogen atmosphere. The product was then filtered through a cellulite filter and purified under reduced pressure. Purification was performed by chromatography (mobile phase: ethyl acetate: hexane = 1: 10 to 1: 4 mixture). The yield of the obtained product was 85%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 1.40-1.50 (1H, m), 1.60-1.80 (4H, m), 1.80-1.90 (1H, m), 3 .20 (1H, s), 3.54 (1H, s), 3.67 (1H, s), 3.79 (1H, s), 3.82-3.88 (4H, m);
¹³ C-NMR (CDCl ₃ ): δ 26.5, 30.1, 37.5, 64.0, 64.2, 69.1, 70.0, 108.6;
IR (KBr): 3417, 2958, 2935, 2888, 1442, 1144, 1101, 1059 cm ⁻¹ ;
[Α] _D ²¹ +2.6 (c = 1.5, CHCl ₃ );
HRMS (FAB): calculated [C ₈ H ₁₄ O ₄ Na]: 197.0785, found: 197.1784.

<Synthesis of 4-hydroxy-cyclohexyl-2-enone>
Synthesis of 4-hydroxy-cyclohexyl-2-enone represented by the structural formula (7) in the above synthesis scheme (A) was performed. The synthesis scheme is as follows.

  The diol (118 mg, 0.677 mmol) represented by the structural formula (6) synthesized by the above method was added to a mixed solution of tetrahydrofuran (9.7 ml), water (0.97 ml) and acetone (0.97 ml). . Further, 24.2 mg of ion exchange resin (trade name: Amberlite 15 (20% by mass)) was added and stirred at 80 ° C. for 15 hours. To the solution after the reaction, anhydrous magnesium sulfate was added for dehydration, and the filtrate was distilled off under reduced pressure. Subsequently, it was purified by chromatography (mobile phase was a mixture of ethyl acetate: hexane = 3: 1). The yield of the obtained product was 85%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 1.94-2.02 (1H, m), 2.30-2.40 (2H, m), 2.40-2.50 (1H, s), 2 .52-2.59 (1H, dt, J = 17.4, 4.7 Hz), 4.54-4.57 (1H, ddd, J = 6.8, 4.6, 2.2 Hz), 5 .93-5.95 (1H, d, J = 10.0 Hz), 6.90-6.93 (1H, dt, J = 10.0, 1.8 Hz);
¹³ C-NMR (CDCl ₃ ): δ 32.4, 35.3, 66.3, 129.2, 152.9, 198.9;
IR (KBr): 3419, 2954, 2871, 1660, 1205, 1066, 970, 943, 864 cm ⁻¹ ;
[Α] _D ³⁰ +92.3 (c = 0.7, CHCl ₃ );
HRMS (FAB): calculated value [C ₆ H ₈ O ₂ ]: 112.0524, actually measured: 112.0531.

<Synthesis of 5- (tertiarybutyl-dimethyl-siloxy) -7-oxa-bicyclo [4.1.0] heptan-2-one>
Synthesis of 5- (tertiarybutyl-dimethyl-siloxy) -7-oxa-bicyclo [4.1.0] heptan-2-one represented by the structural formula (8) in the above synthesis scheme (A) was performed. It was. The synthesis scheme is as follows.

  Dimethylformamide (1.8 ml) in which enone represented by the structural formula (7) synthesized by the above method (100 mg, 0.892 mmol) and tertiary butyldimethylsilyl chloride (TBSCl) (341 mmg, 2.27 mmol) are dissolved. ) Was added imidazole (168 mg, 2.47 mmol), and the mixture was stirred at 0 ° C. for 1 hour. The reaction was then terminated by adding a pH 7 phosphate buffer. Then, ethyl acetate was added to extract the product, and the product was further washed with brine. The washed product was dehydrated by adding anhydrous sodium sulfate. This was filtered and distilled under reduced pressure to purify. Purification was performed by chromatography (mobile phase: ethyl acetate: hexane = 1: 10 to 1: 4 mixture). The yield of the obtained product was 75%.

  Next, the product (ester compound of enone represented by the structural formula (7)) (345 mg, 1.52 mmol) obtained by the above reaction was dissolved in tetrahydrofuran (9.5 ml) at 0 ° C. Aqueous hydrogen oxide (0.88 ml, 7.63 mmol) and benzyltrimethylammonium hydroxide (69 μL, 0.15 mmol) were added and reacted at 0 ° C. for 0.5 hour, followed by addition of an ammonium chloride solution. Was terminated.

  The aqueous phase was then extracted with ethyl acetate. The organic phase was washed with brine and sodium carbonate and dehydrated by adding sodium sulfate. Further, anhydrous sodium sulfate was added to the organic phase for dehydration and distilled off under reduced pressure. Subsequently, it refine | purified using the chromatography (The mobile phase is a mixed liquid of ethyl acetate: hexane = 1: 50). The yield of the obtained product was 76%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
Enone ester compound represented by Structural Formula (7)
¹ H-NMR (CDCl ₃ ): δ 0.01 (3H, s), 0.02 (3H, s), 0.81 (9H, s), 1.90-2.00 (1H, m), 2 10-2.20 (1H, m), 2.26-2.35 (1H, ddd, J = 12.7, 17.0, 4.5 Hz), 2.49-2.56 (1H, dt , J = 16.8, 4.4 Hz), 4.46-4.51 (1H, tt, J = 6.8, 2.1 Hz), 5.86-5.89 (1H, d, J = 10). .1 Hz), 6.77-6.80 (1H, dt, J = 10.2, 1.8 Hz);
¹³ C-NMR (CDCl ₃ ): δ 1.4, 1.4, 18.3, 26.1, 33.3, 35.9, 67.4, 129.1, 154.3, 199.3;
IR (KBr): 2954, 2858, 1691, 1471, 1383, 1252, 1103, 860, 837 cm ⁻¹ ;
[Α] _D ³⁰ +97.9 (c = 1.2, CHCl ₃ );
HRMS (FAB): calculated _{[C 11 H 19 O 2 Si} ] [M-CH 3] +: 211.1154, Found: 211.1186.

Epoxide represented by structural formula (8)
¹ H-NMR (CDCl ₃ ): δ 0.03 (3H, s), 0.04 (3H, s), 0.81 (9H, s), 1.59-1.66 (1H, m), 1 97-2.05 (1H, m), 2.20-2.35 (2H, m), 3.17-3.18 (1H, d, J = 3.9 Hz), 3.37-3. 39 (1H, t, J = 3.1 Hz), 4.36-4.39 (1H, dd, J = 6.8, 3.1 Hz);
¹³ C-NMR (CDCl ₃ ): δ-5.0, -4.9, 17.9, 25.4, 25.5, 31.5, 54.8, 57.9, 65.1;
IR (KBr): 2954, 2858, 1716, 1473, 1362, 1254, 1092, 981, 839, 777 cm ⁻¹ ;
[Α] _D ³⁰ -48.5 (c = 1.0, CHCl ₃ );
HRMS (FAB): calculated _{[C 11 H 19 O 3 Si} ] [M-CH 3] +: 227.1103, Found: 227.1075.

<Synthesis of 5- (tertiarybutyl-dimethyl-siloxy) -7-oxa-bicyclo [4.1.0] hept-3-en-2-one>
5- (tertiarybutyl-dimethyl-siloxy) -7-oxa-bicyclo [4.1.0] hept-3-en-2-one represented by structural formula (9) in the above synthesis scheme (A) Was synthesized. The synthesis scheme is as follows.

  To an acetonitrile solution (2.2 ml) in which epoxide (105 mg, 0.433 mmol) represented by the structural formula (8) synthesized by the above method and triethylamine (0.9 ml, 6.71 mmol) are dissolved, 0 ° C. Originally, trimethylsilyl chloride (TMSCl) (0.8 ml, 6.50 mmol) was added. This was stirred at room temperature for 45 hours to react, and a pH 7 phosphate buffer was added to terminate the reaction. The resulting product was extracted with ethyl acetate.

  The organic phase was then washed with brine and dehydrated by adding sodium sulfate. Further, anhydrous sodium sulfate was added to the organic phase for dehydration and distilled off under reduced pressure. The obtained trimethylsilyl compound was used in the next reaction without purification.

Tris (dibenzylideneacetone) dipalladium (Pd ₂ dba ₃ .CHCl ₃ ) (62.7 mg, 0.06 mmol) and diaryl carbonate (58 μl, 0.40 mmol) were added to an acetonitrile solution of the above product ( 7.2 ml) and stirred at room temperature for 4 hours. Thereafter, a pH 7 phosphate buffer was added to terminate the reaction, and the organic phase was extracted with chloroform. Subsequently, anhydrous sodium sulfate was added to the organic phase for dehydration and distilled off under reduced pressure. Subsequently, it refine | purified using the chromatography (The mobile phase is a mixed liquid of ethyl acetate: hexane = 1: 50). The yield of the obtained product was 78%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 0.12 (3H, s), 0.15 (3H, s), 0.89 (9H, s), 3.42 to 3.42 (1H, m), 3 .60-3.62 (1H, m), 4.62-4.63 (1H, m), 5.94-5.97 (1H, dt, J = 10.5, 1.3 Hz), 6. 50-6.54 (1H, ddd, J = 10.5, 4.5, 2.7 Hz);
¹³ C-NMR (CDCl ₃ ): δ-4.7, -4.5, 18.1, 25.6, 53.3, 53.4, 63.6, 126.2, 144.3, 193. 2;
IR (KBr): 2956, 2931, 2858, 1693, 1261, 1092, 839, 806, 779 cm ⁻¹ ;
[Α] _D ²⁰ -340 (c = 3.20, CH ₂ Cl ₂ );
HRMS (FAB): calculated _{[C 12 H 20 O 3 Si} ]: 240.1182, Found: 240.1090.

<Synthesis of 5- (tertiarybutyl-dimethyl-siloxy) -3-iodo-7-oxa-bicyclo [4.1.0] hept-3-en-2-one>
5- (tertiarybutyl-dimethyl-siloxy) -3-iodo-7-oxa-bicyclo [4.1.0] hept-3-ene represented by the structural formula (10) in the above synthesis scheme (A) 2-one was synthesized. The synthesis scheme is as follows.

  A mixed solution of diethyl ether (1.5 ml) and pyridine (1.5 ml) in which iodo (305 mg, 1.20 mmol) was dissolved was stirred at 0 ° C. in a dark room for 20 minutes. To this reaction solution, enone represented by the structural formula (9) synthesized by the above method (144 mg, 0.600 mmol) was added and stirred at 0 ° C. for 2 hours. Thereafter, a pH 7 phosphate buffer was added to terminate the reaction, and the organic phase was extracted with ethyl acetate. Subsequently, anhydrous sodium sulfate was added to the organic phase for dehydration and distilled off under reduced pressure. Subsequently, purification was performed using chromatography (mobile phase was a mixed solution of ethyl acetate: hexane = 1: 50) to obtain a novel compound and a panepophenanthrin derivative according to the present invention. The yield of the obtained derivative was 80%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 0.16 (3H, s), 0.18 (3H, s), 0.92 (9H, s), 3.62-3.63 (1H, m), 3 68-3.69 (1H, m), 4.59-4.60 (1 H, m), 7.27-7.29 (1 H, dd, J = 5.0, 2.4 Hz);
¹³ C-NMR (CDCl ₃ ): δ-4.7, -4.5, 18.1, 25.6, 51.7, 58.2, 66.1, 101.9, 152.6, 187. 5;
IR (KBr): 2954, 2858, 1697, 1257, 1092, 872, 835, 781 cm ⁻¹ ;
^{_{[Α] D 22 -105.6 (c}} = 1.2, CHCl 3);
HRMS (FAB): calculated _{[C 12 H 20 IO 3 Si} ]: 367.0227, Found: 367.0249.

[Synthesis of Panepophenanthrin]
Synthesis of <5- (tertiarybutyl-dimethyl-siloxy) -3- (3-hydroxy-3-methyl-butynyl) -7-oxa-bicyclo [4.1.0] hept-3-en-2-one >
5- (tertiarybutyl-dimethyl-siloxy) -3- (3-hydroxy-3-methyl-butynyl) -7-oxa-bicyclo [4] represented by the structural formula (12) in the above synthesis scheme (A) .1.0] Hept-3-en-2-one was synthesized. The synthesis scheme is as follows.

Tris (dibenzylideneacetone) dipalladium (Pd ₂ dba ₃ .CHCl ₃ ) (8.2 mg, 0.01 mmol) and triphenylarsine (7.8 mg, 0.03 mmol) were mixed under reduced pressure, 0.2 ml Toluene was further added and stirred at room temperature for 20 minutes. To this reaction solution, a derivative (144 mg, 0.600 mmol) represented by the structural formula (10) synthesized by the above method was added, and vinyl stannane (37.6 mg, 0.1 mmol) represented by the structural formula (11) was added. ) Was added and stirred at 110 ° C. for 5 minutes. Thereafter, the inorganic product was removed by filtration, and the filtrate was distilled off under reduced pressure. Subsequently, it refine | purified using the chromatography (The mobile phase is a mixed liquid of ethyl acetate: hexane = 1: 5). The yield of the obtained derivative was 77%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 0.13 (3H, s), 0.16 (3H, s), 0.90 (9H, s), 1.33 (3H, s), 1.35 (3H , S), 3.41-3.51 (1H, d, J = 3.3 Hz), 3.60-3.65 (1H, m), 4.71-4.73 (1H, d, J = 4.9 Hz), 6.26-6.30 (1 H, d, J = 16.1 Hz), 6.38 (1 H, s), 6.38-6.43 (1 H, d, J = 16.1 Hz) );

Panepophenanthrin represented by the structural formula (1) in the above synthesis scheme was synthesized. The synthesis scheme is as follows.

Excess ammonium fluoride was added to 3 ml of a methanol solution (18.9 mg, 0.06 mmol) of the compound represented by the structural formula (12) synthesized by the above method, followed by stirring at room temperature for 12 hours. The reaction was concentrated under reduced pressure. The residue was purified by thin layer chromatography to obtain a monomer (9.3 mg, 0.04 mmol) represented by the following structural formula (2). The yield of this compound was 75%. This monomer was allowed to stand at room temperature for 33 hours to obtain panepophenanthrin (9.2 mg, 0.02 mmol). The yield at this time was 95%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
Monomer represented by (2)
¹ H-NMR (CDCl ₃ ): δ 1.04 (3H, s), 1.05 (3H, s), 3.30-3.31 (1H, d, J = 3.5 Hz), 3.53- 3.55 (1H, m), 4.49-4.50 (1H, d, J = 5.1 Hz), 6.03-6.07 (1H, d, J = 16.1 Hz), 6.16 −6.20 (1H, d, J = 16.1 Hz), 6.31-6.33 (1H, dd, J = 5.1, 2.4 Hz);

Panepophenanthrin
¹ H-NMR (CDCl ₃ ): 1.17 (3H, s), 1.20 (3H, s), 1.35 (3H, s), 1.45 (3H, s), 2.03 (1H , Brd, J = 9.7 Hz), 2.32 (1H, brd, J = 10.0 Hz), 3.31 (1H, d, J = 4.0 Hz), 3.35 (1H, dd, J = 5.0, 1.6 Hz), 3.42 (1 H, d, J = 4 Hz), 3.50 (1 H, t, J = 3.2 Hz), 3.84 (1 H, t, J = 3.4 Hz) ), 4.35 (1 H, br s), 4.55 (1 H, br s), 5.68 (1 H, d, J = 16.2 Hz), 5.99 (1 H, d, J = 16.2 Hz). ), 6.81 (1H, dd, J = 5.0, 3.0 Hz);
¹³ C-NMR (CDCl ₃ ): δ 26.2, 29.5, 30.3, 32.3, 50.0, 51.2, 55.1, 55.60.7, 66.2, 69.0 71.8, 79.2, 102.7, 129.3, 138.8, 139.9, 143.0, 196.3;
IR (KBr): 2978, 1676, 1597, 1338, 1142, 997 cm ⁻¹ ;
[Α] _D ²⁴ +147.2 (c = 0.91, MeOH);
HRMS (FAB): calculated _{[C 22 H 27 O 8]} : 419.1719, Found: 419.1706.

[Synthesis of Novel Compound (1)]
<Synthesis of 5- (tertiarybutyl-dimethyl-siloxy) -3-propenyl-7-oxa-bicyclo [4.1.0] hept-3-en-2-one>
5- (tertiarybutyl-dimethyl-siloxy) -3-propenyl-7-oxa-bicyclo [4.1.0] hept-3-ene represented by the structural formula (16) in the above synthesis scheme (A) 2-one was synthesized. The synthesis scheme is as follows.

The derivative (23.5 mg, 0.06 mmol) represented by the structural formula (10) synthesized by the above method, the compound (10.3 mg, 0.12 mmol) represented by the structural formula (13), silver oxide (23.8 mg) 0.1 mmol) and AsPh3 (11.8 mg, 0.04 mmol) in a THF-water mixture (8: 1, 1.4 ml), tris (dibenzylideneacetone) dipalladium (Pd ₂ dba ₃ .CHCl ₃ ) (7.4 mg, 0.02 mmol) was added, and the mixture was stirred at room temperature / dark room for 30 minutes. To this reaction solution was added 5 ml of ammonium chloride solution, and the mixture was further stirred for 1 hour. The purified product was extracted with ethyl acetate, sodium sulfate was added, and the mixture was distilled off under reduced pressure. The organic phase was concentrated under reduced pressure and then purified by thin layer chromatography to obtain a siloxy monomer (15.5 mg, 0.055 mmol) represented by the structural formula (16). The yield at this time was 92%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 0.12 (3H, s), 0.15 (3H, s), 0.90 (9H, s), 1.78-1.80 (3H, d, J = 6.4 Hz), 3.41-3.42 (1 H, d, J = 4.0 Hz), 3.62 (1 H, s), 4.70-4.71 (1 H, d, J = 4.0 Hz) ), 6.08-6.12 (1 H, d, J = 16.2 Hz), 6.19-6.26 (1 H, m), 6.30-6.40 (1 H, m);

Next, a novel compound represented by the structural formula (20) was synthesized. The synthesis scheme is as follows.

Excess ammonium fluoride was added at room temperature to 3 ml of methanol in which the siloxy monomer (15.5 mg, 0.055 mmol) represented by the structural formula (16) synthesized by the above method was dissolved. After stirring for 12 hours, the reaction solution was concentrated under reduced pressure. The residue was purified by thin layer chromatography to obtain a monomer (9.3 mg, 0.04 mmol) represented by the following structural formula (16 ′). The yield of this compound was 75%. The monomer was allowed to dimerize at room temperature for 5 hours to obtain a novel compound (15.0 mg, 0.05 mmol) represented by the structural formula (20). The yield at this time was 80%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 0.83-0.91 (3H, d, J = 7.2 Hz), 1.66 to 1.68 (3H, dd, J = 6.5, 1.5 Hz) , 2.35-2.38 (1H, m), 2.60-2.70 (1H, m), 2.90-3.00 (1H, m), 3.22-3.23 (1H, d, J = 3.4 Hz), 3.28-3.29 (1H, d, J = 3.4 Hz), 3.49-3.50 (1H, d, J = 3.4 Hz), 3.51 -3.52 (1H, d, J = 3.4Hz), 3.90-3.92 (1H, d, J = 9.0Hz), 4.57-4.58 (1H, d, 4.2Hz) ), 5.20-5.40 (1H, m), 5.54-5.58 (1H, dd, J = 16.2, 1.5 Hz), 6.59-6.61 (1H, dd, J = 4.7, 2.3Hz)
¹³ C-NMR (CDCl ₃ ): δ 15.2, 18.4, 29.6, 34.5, 44.8, 53.3, 53.6, 58.1, 61.9, 66.5 70.6, 76.4, 129.1, 131.0, 131.8, 142.6, 195.4, 203.0;
IR (KBr): 3419, 2923, 2854, 1698, 1633, 1455, 1259, 1085 cm ⁻¹ ;
[Α] _D ²² +83.1 (c = 0.1, CHCl ₃ );
HRMS (FAB): calculated value [C ₁₈ H ₂₁ O ₆ ]: 333.1338, actual value: 333.1357.

[Synthesis of Novel Compound 2]
<Synthesis of 5- (tertiarybutyl-dimethyl-siloxy) -3-hexyl-7-oxa-bicyclo [4.1.0] hept-3-en-2-one>
5- (tertiarybutyl-dimethyl-siloxy) -3-hexyl-7-oxa-bicyclo [4.1.0] hept-3-ene represented by the structural formula (17) in the above synthesis scheme (A) 2-one was synthesized. The synthesis scheme is as follows.

The derivative represented by the structural formula (10) (3.7 mg, 0.01 mmol), the compound represented by the structural formula (14) (2.6 mg, 0.02 mmol), silver oxide (3. 7 mg, 0.02 mmol) and triphenylarsine (1.8 mg, 0.01 mmol) in a THF-water mixture (8: 1, 0.4 ml) were mixed with tris (dibenzylideneacetone) dipalladium (Pd ₂ dba _3. CHCl ₃ ) (1.2 mg, 0.003 mmol) was added, and the mixture was stirred at room temperature / dark room for 1 hour. To this reaction solution was added 2 ml of ammonium chloride solution, and the mixture was further stirred for 1 hour. The purified product was extracted with ethyl acetate, sodium sulfate was added, and the mixture was distilled off under reduced pressure. The organic phase was concentrated under reduced pressure and then purified by thin layer chromatography to obtain a siloxy monomer (2.3 mg, 0.007 mmol) represented by the structural formula (17). The yield at this time was 70%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 0.13 (3H, s), 0.15 (3H, s), 0.90 (9H, s), 1.21-1.40 (9H, m), 3 .50-3.51 (1H, d, J = 4.4 Hz), 3.62 (1H, s), 4.70-4.72 (1H, d, J = 4.4 Hz), 6.06- 6.10 (1H, d, J = 16.1 Hz), 6.18-6.32 (1 H, m), 6.30-6.37 (1 H, m);

Next, a novel compound represented by the structural formula (21) was synthesized.

Excess ammonium fluoride was added at room temperature to 0.6 ml of a methanol solution of the siloxy monomer (2.3 mg, 0.007 mmol) represented by the structural formula (17) obtained by the above method and stirred for 12 hours. went. Thereafter, the product was concentrated under reduced pressure and purified by thin layer chromatography to obtain a monomer represented by the following structural formula (17 ′). This monomer was allowed to stand for 5 hours to obtain a dimer (1.2 mg, 0.003 mmol) represented by the structural formula (21). The yield of this dimer was 83%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ 0.80-0.90 (6H, m), 1.20-1.28 (12H, m), 2.43-2.50 (1H, dd, J = 6) 0.0, 3.8 Hz), 2.72-2.79 (1H, m), 2.80-2.87 (1H, m), 3.28-3.29 (1H, d, J = 3. 3 Hz), 3.32-3.33 (1H, d, J = 3.3 Hz), 3.52-3.53 (1H, d, J = 3.3 Hz), 3.55-3.57 (1H , D, J = 3.3 Hz), 4.01-4.04 (1H, d, J = 9.2 Hz), 4.70-4.80 (1H, d, J = 3.8 Hz), 5 27-5.33 (1H, td, 6.8, J = 16.4 Hz), 5.57-5.61 (1H, d, J = 16.4 Hz), 6.77-6.79 (1H , Dd, J = 2.8,2. 1 Hz);
¹³ C-NMR (CDCl ₃ ): δ 13.8, 22.2, 22.6, 29.5, 29.7, 30.4, 31.0, 32.6, 39.9, 44.6, 47 0.0, 53.3, 53.9, 57.5, 61.7, 67.4, 71.6, 130.1, 131.6, 134.8, 142.3, 194.5, 202.4 ;
IR (KBr): 3444, 2927, 2857, 1698, 1635, 1465, 1268 cm ⁻¹ ;
[Α] _D ²¹ +25.8 (c = 0.2, CHCl 3);
HRMS (FAB): calculated [C ₂₄ H ₃₃ O ₆ ]: 417.2277, found: 417.2265.

[Synthesis of Novel Compound 3]
<Synthesis of 5- (tertiarybutyl-dimethyl-siloxy) -3-dec-1-enyl-7-oxa-bicyclo [4.1.0] hept-3-en-2-one>
5- (tertiarybutyl-dimethyl-siloxy) -3-dec-1-enyl-7-oxa-bicyclo [4.1.0] hept represented by the structural formula (18) in the above synthesis scheme (A) Synthesis of -3-en-2-one was performed. The synthesis scheme is as follows.

The derivative represented by the structural formula (10) (3.7 mg, 0.01 mmol), the compound represented by the structural formula (15) (3.7 mg, 0.02 mmol), silver oxide (3. 7 mg, 0.02 mmol) and triphenylarsine (1.8 mg, 0.01 mmol) in a THF-water mixture (8: 1, 0.4 ml) were mixed with tris (dibenzylideneacetone) dipalladium (Pd ₂ dba _3. CHCl ₃ ) (1.2 mg, 0.003 mmol) was added, and the mixture was stirred at room temperature / dark room for 1 hour. To this reaction solution was added 2 ml of ammonium chloride solution, and the mixture was further stirred for 1 hour. The purified product was extracted with ethyl acetate, sodium sulfate was added, and the mixture was distilled off under reduced pressure. The organic phase was concentrated under reduced pressure and then purified by thin layer chromatography to obtain a siloxy monomer (2.5 mg, 0.0067 mmol) represented by the structural formula (18). The yield at this time was 67%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): 0.12 (3H, s), 0.15 (3H, s), 1.00 (9H, s), 1.15-1.30 (17H, m), 3 .48-3.50 (1H, d, J = 4.1 Hz), 3.62 (1H, s), 4.70-4.72 (1H, d, J = 4.1 Hz), 6.06- 6.10 (1H, d, J = 15.8 Hz), 6.17-6.28 (1 H, m), 6.30-6.37 (1 H, m);

Next, a novel compound represented by (22) was synthesized.

Excess ammonium fluoride was added at room temperature to 0.6 ml of a methanol solution of the siloxy monomer (2.5 mg, 0.0067 mmol) represented by the structural formula (18) obtained by the above method and stirred for 12 hours. went. Thereafter, the product was concentrated under reduced pressure and purified by thin layer chromatography to obtain a monomer represented by the following structural formula (18 ′). This monomer was allowed to stand for 5 hours to obtain a dimer (1.5 mg, 0.003 mmol) represented by the structural formula (22). The yield of this dimer was 84%.

The results of ¹ H-NMR, ¹³ C-NMR, IR and the like of the product are shown below.
¹ H-NMR (CDCl ₃ ): δ0.84-0.87 (6H, m), 1.00-1.06 (28H, m), 2.45-2.47 (1H, dd, J = 6 .3, 4.0 Hz), 2.72-2.80 (1H, m), 2.80-2.86 (1H, m), 3.27-3.28 (1H, d, J = 3. 3 Hz), 3.31-3.32 (1H, d, J = 3.3 Hz), 3.52-3.53 (1H, d, J = 3.3 Hz), 3.54-3.56 (1H , D, J = 3.3 Hz), 4.01-4.04 (1H, d, J = 9.2 Hz), 4.70-4.80 (1H, d, J = 4.0 Hz), 5. 27-5.33 (1H, td, J = 6.9, 16.3 Hz), 5.56-5.60 (1H, d, J = 16.3 Hz), 6.77-6.78 (1H, dd, J = 5.0,2. Hz);
¹³ C-NMR (CDCl ₃ ): δ 14.1, 22.6, 27.4, 28.9, 29.2, 29.3, 29.4, 29.6, 29.7, 30.8, 31 8, 33.0, 40.0, 44.7, 47.1, 53.3, 53.5, 53.9, 57.5, 61.8, 67.5, 71.7, 130.2 , 131.6, 134.9, 142.2, 194.5, 202.5;
IR (KBr): 3434, 2925, 2854, 1702, 1465, 1270, 1054 cm ⁻¹ ;
[Α] _D ²³ +33.8 (c = 0.1, CHCl ₃ );
HRMS (FAB): calculated [C ₃₂ H ₄₉ O ₆ ]: 529.3529, found: 529.3504.

[Example 1]
The ubiquitin activating enzyme (E1 enzyme) inhibitory activity of the novel compound according to the present invention was examined.

In a reaction solution (100 mM, Tris-HCl: pH 9.0, 5 mM magnesium chloride, 1 mM dithiothreitol, 25 mM adenosine triphosphate (ATP)) containing recombinant E1 enzyme (10 μg / ml, manufactured by Boston Biochemical) The novel compounds (a), (b) and (c) according to the present invention having the following structural formula were added, respectively, and allowed to stand at room temperature for 15 minutes.

  Subsequently, biotinylated ubiquitin (100 ng / 1 reaction, manufactured by Boston Biochemical Co.) was added and reacted at 37 ° C. for 15 minutes. Thereafter, loading buffer (loading buffer) (final concentration: 62.5 mM, Tris-HCl: pH 6.8, 2% sodium dodecyl sulfate (SDS), 10% glycol) was added and treated at 100 degrees for 3 minutes. did. Then, the ubiquitinated E1 enzyme is separated by 7.5% SDS-PAGE (SDS-polyacrylamide gel electrophoresis), and is applied to a polyvinylidene fluoride (PVDF) membrane (manufactured by Nihon Millipore) by a semi-dry method. After the transfer, blocking and washing were performed.

Next, the PVDF membrane was treated with a streptavidin-conjugated HRP solution, and then ubiquitinated E1 enzyme was detected with a chemiluminescence kit (trade name Super Signal West Pico; manufactured by Pierce). The band indicating the ubiquitinated E1 enzyme on the gel was quantified by scanning imager (manufactured by Molecular Dynamics). As a result, the 50% inhibitory concentrations (IC ₅₀ values) of the E1 enzyme of the novel compounds (a), (b), and (c) according to the present invention were 9.4, 3.5, and 90 μM, respectively. It was. In addition, the IC ₅₀ value of panepophenanthrin was 70 μM.

[Example 2]
The antitumor activity of the novel compound according to the present invention was examined.

Human breast cancer cells MCF-7 cells ^{(3 × 10 3 / 200μl)} , were cultured in 96-well plastic dishes containing RPMI-1640 medium (containing 10% fetal calf serum). The novel compounds (a), (b) and (c) according to the present invention were added thereto, and after culturing at 37 ° C. in a 5% carbon dioxide atmosphere for 48 hours, a reagent for measuring the number of cells (trade name) : WST-8 reagent [2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2,4-disulphophenyl) -2H-tetrazoleum, monosodium salt]) was added, and further 3 hours After culturing, the absorbance at 450 nm was measured to calculate the survival rate.

As a result, the 50% growth inhibitory concentrations (IC ₅₀ values) of the novel compounds (a), (b), and (c) according to the present invention were 5.4, 1.0, and 3.6 μM, respectively. It was. In addition, the IC ₅₀ value of panepophenanthrin was 100 μM. From this, it was shown that the novel compounds (a), (b) and (c) according to the present invention are effective as antitumor agents.

Claims (6)

A novel compound having a structure represented by the general formula (1).

[Wherein R ₁ has a hydrogen atom, an alkyl group having 1 to 30 carbon atoms which may have a functional group, an aryl group having 6 to 12 carbon atoms which may have a substituent, or a substituent. And any one group selected from the group consisting of heteroaryl groups having 6 to 12 carbon atoms. ]   A ubiquitination inhibitor comprising the novel compound according to claim 1 as an active ingredient.   A pharmaceutical comprising the ubiquitination inhibitor according to claim 2 as an active ingredient. A method for producing a novel compound having a structure represented by the general formula (1),
A first reaction step in which a carbonyl compound and a nitroso compound are reacted in the presence of an asymmetric catalyst;
A second reaction step in which the product produced by the first reaction step is stereoselectively reacted to obtain a derivative of the novel compound;
The manufacturing method of the novel compound which has a structure shown by General formula (1) which has this.

[Wherein R ₁ has a hydrogen atom, an alkyl group having 1 to 30 carbon atoms which may have a functional group, an aryl group having 6 to 12 carbon atoms which may have a substituent, or a substituent. And any one group selected from the group consisting of heteroaryl groups having 6 to 12 carbon atoms. ]   The method for producing a novel compound according to claim 4, wherein the asymmetric catalyst is proline. The second reaction step includes a reduction step of reductively cleaving the aminooxy group in the product obtained in the first reaction step to a hydroxyl group;
An epoxidation step of epoxidizing the hydroxyl group;
A side chain introduction step for introducing side chains into the epoxy compound obtained by this epoxidation step;
The method for producing a novel compound according to claim 4 or 5, wherein the method comprises: