KR102619484B1 - Method for preparing DHPV and its enantiomer, and uses of DHPV produced thereof - Google Patents

Abstract

This relates to a method for producing DHPV and its optical isomers, and the use of DHPV produced by the method. According to the method, not only can DHPV be synthesized in large quantities, but optical isomers can be selectively synthesized, thereby improving the biological activity of the isomers. It can be used for various purposes. Additionally, DHPV prepared by the above method can be used to prevent or treat inflammatory bowel disease by regulating the NF-κB signaling pathway.

Description

Method for preparing DHPV and its enantiomer, and uses of DHPV produced by the method {Method for preparing DHPV and its enantiomer, and uses of DHPV produced thereof}

It relates to a method for producing DHPV and its optical isomers, and to the use of DHPV produced by the method.

Inflammatory bowel disease (IBD) is a chronic and multifactorial inflammatory condition of the gastrointestinal tract associated with infectious and immunological imbalances in the intestinal mucosa. The incidence and prevalence of inflammatory bowel disease have continuously increased over the past decades, and it is known to be one of the most important risk factors for the development of colorectal cancer. Inflammatory bowel disease is generally divided into two main types: Crohn's disease (CD) and ulcerative colitis (UC). Crohn's disease can involve any part of the gastrointestinal tract and causes inflammation of entire non-contiguous layers, while ulcerative colitis occurs in the lining of the large intestine or rectum and causes persistent inflammation of the colonic mucosa. The disease causes immunity, damage and symptoms such as diarrhea and bloody stool. These responses are controlled by NF-κB, an immune response regulator. Patients with inflammatory bowel disease are known to have dysregulation of NF-κB signaling, which is associated with rapid and vigorous production of various pro-inflammatory mediators.

Meanwhile, catechin and epicatechin undergo intestinal metabolism to produce various metabolites such as caffeic acid, ferulic acid, and isoferulic acid. Among the various metabolites produced from polyphenols in the human body, 5-(3',4'-dihydroxyphenyl)-γ-valerolactone [(5-(3',4-dihydroxyphenyl)-γ-valerolactone, DHPV ) is a representative product of flavonols containing catechin and epicatein moieties. DHPV has antioxidant activities such as UVB-induced wrinkle formation and brain neuroprotection effects, and can exist as two optical isomers due to the position of the stereocenter of butyrolactone. However, in numerous literature, their structures are expressed ambiguously or in racemic form, and completely different biological activities of optical isomers are reported.

Therefore, based on the biological properties of the optical isomers of DHPV, it is necessary to establish a new treatment strategy for inflammatory bowel disease.

One aspect is to provide a method for producing DHPV (5-(3',4'-dihydroxyphenyl)-γ-valerolactone).

Another aspect is to provide DHPV (5-(3',4'-dihydroxyphenyl)-γ-valerolactone) prepared by the above method.

Another aspect is providing a pharmaceutical composition for the prevention or treatment of inflammatory bowel disease containing DHPV (5-(3',4'-dihydroxyphenyl)-γ-valerolactone) or a pharmaceutically acceptable salt thereof as an active ingredient. will be.

Another aspect is providing a health functional food composition for preventing or improving inflammatory bowel disease containing DHPV (5-(3',4'-dihydroxyphenyl)-γ-valerolactone) or a foodologically acceptable salt thereof as an active ingredient. It is done.

One aspect includes synthesizing a compound represented by Formula 6 below through a cross metathesis (CM) reaction of a compound represented by Formula 4 below and a compound represented by Formula 5 below; Synthesizing a compound represented by Formula 2 below by subjecting the compound represented by Formula 6 synthesized through the above reaction to a Sharpless Asymmetric Dihydroxylation (SAD) reaction; Synthesizing a compound represented by Formula 7 below through hydrogenolysis of the hydroxyl group of the compound represented by Formula 2 synthesized through the above reaction; And a method for producing DHPV (5-(3',4'-dihydroxyphenyl)-γ-valerolactone) comprising the step of deprotecting the compound represented by the following formula (7) synthesized through the above reaction:

[Formula 4]

[Formula 5]

[Formula 6]

[Formula 2]

[Formula 7]

In the above,

X and R ₁ are each hydrogen (H) and an alkyl group of C ₁ to C ₄ ;

R ₂ is each hydrogen (H) and an alkoxy group of C ₁ to C ₄ .

The previously known synthesis method of DHPV is inefficient because it requires several steps, and there was a problem in that it was difficult to mass-produce it effectively in the processing stage by using optically active starting materials. However, according to the method according to one aspect, it is not only efficient because it goes through four relatively simple steps, but also has the advantage of enabling mass production of DHPV because optical isomers can be selectively synthesized.

Figure 2a shows a manufacturing method of DHPV according to one aspect.

The method may include synthesizing a compound represented by Formula 6 through a cross metathesis (CM) reaction of the compound represented by Formula 4 and the compound represented by Formula 5 (FIG. 2b). .

In one embodiment, the compound represented by Formula 4 is 4-(1-propenyl)-1,2-methoxybenzene (4-(1-propenyl)-1,2-dimethoxybenzene) or 4-(1 It may be -propenyl)-1,2-ethoxybenzene (4-(1-propenyl)-1,2-dimethoxybenzene). Additionally, the compound represented by Formula 5 may be ethyl pent-4-enoate or methyl pent-4-enoate. In addition, the compound represented by 6 is (E)-methyl-5-(3,4-dimethoxyphenyl)pent-4-enoate [(E)-Ethyl-5-(3,4-deimethoxyphenyl)pent- 4-enoate] or (E)-ethyl-5-(3,4-dimethoxyphenyl)pent-4-enoate [(E)-Ethyl-5-(3,4-deimethoxyphenyl)pent-4-enoate] It may be.

As used herein, the term "cross methathesis (CM) reaction" refers to an asymmetrically substituted alkene through rearrangement of the carbon-carbon double bond from a diene using a catalyst containing a transition metal. ) It is a reaction that synthesizes a compound. Due to its chemical stability, the catalyst does not affect other functional groups except alkene and is used to increase the reactivity of the metathesis reaction. The catalyst includes a Grubbs catalyst or a Schrock catalyst. The Grubbs catalyst includes, for example, Grubbs 1st generation, Grubbs 2nd generation, and Hoveyda-Grubbs 2nd generation catalysts. The catalyst may be used in an amount of 0.01 to 10 equivalents per equivalent of the compound represented by Formula 5. The catalyst may be used in an amount of, for example, 0.01 to 10 equivalents, 0.01 to 8 equivalents, 0.01 to 5 equivalents, 0.02 to 4 equivalents, 0.02 to 2 equivalents, or 0.04 to 1 equivalents, based on 1 equivalent of the compound represented by Formula 5. . At this time, if the capacity of the catalyst is less than the above range, there is a problem that the effect of improving the reaction rate due to the catalyst is not sufficiently exerted, and if it exceeds the above range, there is a problem that the cost is excessive.

In another embodiment, the compound represented by Formula 4 may be used in an amount of 0.1 equivalent or more per 1 equivalent of the compound represented by Formula 5. The compound represented by Formula 4 is, for example, 0.1 equivalent or more, 1 equivalent or more, 1.5 equivalent or more, 2 equivalents or more, 2.5 equivalents or more, 5 equivalents or more, 7 equivalents or more, based on 1 equivalent of the compound represented by Formula 5. Alternatively, it may be used in an amount of 10 equivalents or more. For example, 0.1 to 10 equivalents, 0.1 to 8 equivalents, 0.1 to 6 equivalents, 0.1 to 4 equivalents, 1 to 10 equivalents, 1 to 9 equivalents, 1 to 7 equivalents, 1 to 5 equivalents, 1.5 to 7 equivalents or 1.5 equivalents. It can be used in amounts ranging from 3 to 3 equivalents. At this time, when the dosage of the compound represented by Formula 4 is less than the above range, there is a problem that the yield of the compound represented by Formula 6 is lowered due to homodimerization of the compound represented by Formula 4.

In addition, the method includes the step of synthesizing a compound represented by Formula 2 below through a Sharpless Asymmetric Dihydroxylation (SAD) reaction of the compound represented by Formula 6 synthesized through the above reaction (FIG. 2c) ) may include.

As used herein, the term "Sharpless Asymmetric Dihydroxylation (SAD) reaction" is a form of enantioselective synthesis of compounds that favor the formation of optical or diastereomers using an asymmetric catalyst. It's a method. The asymmetric catalyst includes, for example, AD-mix-α or AD-mix-β, and depending on the type of catalyst used in the reaction, (S)- or (R)-type optical isomers can be synthesized. . In one embodiment, the compound represented by Formula 2 is (S)-5((S)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one[ (S)-5-((S)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one], (R)-5((R)-(3,4-dimethoxy Phenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one [(R)-5-((R)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one ], (S)-5((S)-(3,4-diethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one[(S)-5-((S)-( 3,4-diethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one], (R)-5((R)-(3,4-diethoxyphenyl)(hydroxy)methyl)dihydrofuran- It may be 2(3H)-one [(R)-5-((R)-(3,4-diethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one] or a mixture thereof.

In addition, the method may include the step of synthesizing the compound represented by Formula 7 through hydrogenolysis of the hydroxyl group of the compound represented by Formula 2 synthesized through the reaction (FIG. 2d). . In one embodiment, the compound represented by Formula 7 is (S)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one [(S)-5-(3,4 -dimethoxybenzyl)dihydrofuran-2(3H)-one], (R)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one[(R)-5-(3,4- dimethoxybenzyl)dihydrofuran-2(3H)-one], (S)-5-(3,4-diethoxybenzyl)dihydrofuran-2(3H)-one[(S)-5-(3,4-diethoxybenzyl )dihydrofuran-2(3H)-one], (R)-5-(3,4-diethoxybenzyl)dihydrofuran-2(3H)-one[(R)-5-(3,4-diethoxybenzyl) dihydrofuran-2(3H)-one] or a mixture thereof.

Additionally, the method may include the step of deprotecting the compound represented by the following formula (7) synthesized through the above reaction (FIG. 2e). In one embodiment, the step includes boron tribromide (BBr ₃ ), thiophenol (PHSH), lithium diode (LiI), methylmagnesium iodide (MeMgI), lithium chloride (LiCl), and trimethylsilyl diode (TMSI). ), aluminum tribromide (AlBr ₃ ), aluminum trichloride (AlCl ₃ ), boron triode (BI ₃ ), boron trifluoride (BF ₃ ), and boron trichloride (BCl ₃ ). It can be carried out using one or more catalysts. The catalyst may be used in an amount of 0.01 to 50 equivalents per equivalent of the lactone compound. The catalyst is used, for example, 0.01 to 20 equivalents, 0.01 to 18 equivalents, 0.01 to 16 equivalents, 0.01 to 14 equivalents, 0.01 to 10 equivalents, 1 to 20 equivalents, 1 to 15 equivalents, 3 to 1 equivalent, based on 1 equivalent of the lactone compound. It can be used at 13 equivalents, 4 to 10 equivalents or 4.5 to 8 equivalents. At this time, if the content of the catalyst for performing deprotection is less than the above range, there is a problem that the deprotection reaction is not sufficiently achieved, and if it exceeds the above range, excessive heat generation occurs at the end of the reaction (work-up). There is a problem that arises.

Another aspect provides DHPV (5-(3',4'-dihydroxyphenyl)—γ-valerolactone) prepared by the above method. In one embodiment, the optical purity of the DHPV may be 90% or more and less than 100%. In one example, DHPV prepared by the above method was confirmed through chiral HPLC experiment, and it was confirmed that (S)-DHPV and (R)-DHPV had enantioselectivities of 97.3:2.7 and 95.1:4.9, respectively. . Therefore, the DHPV may be manufactured from optical isomers, and may be (S)-DHPV represented by the following Chemical Formula 1a or (R)-DHPV represented by the following Chemical Formula 1b:

[Formula 1a]

[Formula 1b]

.

Additionally, in one example, as a result of confirming the anti-inflammatory activity of DHPV prepared by the above method, it was confirmed that (S)-DHPV was more effective in suppressing NF-κB signaling than (R)-DHPV. Therefore, the (S)-DHPV can be used as a new therapeutic agent for the prevention and treatment of inflammatory bowel disease.

Another aspect provides a pharmaceutical composition for preventing or treating inflammatory bowel disease containing DHPV (5-(3',4'-dihydroxyphenyl)- γ-valerolactone) of the following formula 1 as an active ingredient:

[Formula 1]

.

In addition, a method of treating inflammatory bowel disease is provided, comprising administering a composition containing DHPV (5-(3',4'-dihydroxyphenyl) - γ-valerolactone) of the following formula (1) to an individual in need thereof: :

[Formula 1]

.

The specific details of the DHPV are as described above. The inflammatory bowel disease may be, for example, enteritis, ulcerative colitis, Chrohn's disease, intestinal Bechet's disease, bleeding rectal ulcer, and ileal pouchitis. In one example, it was confirmed that the DHPV inhibits NF-κB signaling by inhibiting the phosphorylation and degradation of IκBα in rat intestinal epithelial cells stimulated with LPS. Accordingly, the composition according to one aspect can be used to prevent or treat inflammatory bowel disease by regulating the NF-κB signaling pathway.

Pharmaceutical compositions for preventing or treating inflammatory bowel disease according to one aspect are prepared in oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods. It can be formulated and used, and may include appropriate carriers, excipients, or diluents commonly used in the manufacture of pharmaceutical compositions for formulation.

The carrier, excipient or diluent includes lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, not yet determined. These include various compounds or mixtures including quality cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

When formulating, it can be manufactured using diluents or excipients such as commonly used fillers, weighting agents, binders, wetting agents, disintegrants, and surfactants.

Solid formulations for oral administration can be prepared by mixing the DHPV with at least one excipient, such as starch, calcium bonate, sucrose or lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used.

Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives. .

Preparations for parenteral administration include sterile aqueous solutions, non-aqueous preparations, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurel, glycerol gelatin, etc. can be used.

The preferred dosage of the pharmaceutical composition for preventing or treating inflammatory bowel disease according to one aspect varies depending on the patient's condition, body weight, degree of disease, drug form, administration route and period, but may be appropriately selected by a person skilled in the art. However, for desirable effects, it can be administered at 0.0001 to 2,000 mg/kg per day, preferably at 0.001 to 2,000 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. However, the scope of the present invention is not limited by the above dosage.

Pharmaceutical compositions for preventing or treating inflammatory bowel disease according to one aspect may be administered to mammals such as rats, mice, livestock, and humans through various routes. Any mode of administration can be administered, for example, orally, rectally or by intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebroventricular injection.

Another aspect provides a health functional food composition for preventing or improving inflammatory bowel disease containing DHPV (5-(3',4'-dihydroxyphenyl)- γ-valerolactone) of the following formula 1 as an active ingredient:

[Formula 1]

.

In the health functional food composition for improving inflammatory bowel disease according to one aspect, when the DHPV is used as an additive to a health functional food, it can be added as is or used with other foods or food ingredients, and can be added according to a conventional method. It can be used appropriately. The amount of active ingredients mixed can be appropriately determined depending on each purpose of use, such as prevention, health, or treatment.

The formulation of health functional foods can be in the form of powders, granules, pills, tablets, capsules, as well as general foods or beverages.

There are no particular restrictions on the type of food, and examples of foods to which the above substance can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, etc., and can include all foods in the conventional sense.

In general, when manufacturing food or beverages, the DHPV can be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on 100 parts by weight of raw materials. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range. Among health functional foods according to one aspect, beverages may contain various flavoring agents or natural carbohydrates as additional ingredients like regular beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the beverage according to the present invention.

In addition to the above, the health functional food composition for preventing or improving inflammatory bowel disease according to one aspect includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, May contain pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonating agents used in carbonated beverages. In addition, the composition for improving sleep of the present invention may contain pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients can be used independently or in combination. The ratio of these additives is not limited, but is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food of the present invention.

According to the DHPV synthesis method according to one aspect, not only can DHPV be synthesized in large quantities, but also optical isomers can be selectively synthesized, so it can be used for various purposes depending on the biological activity of the isomers. Additionally, DHPV prepared by the above method can be used to prevent or treat inflammatory bowel disease by regulating the NF-κB signaling pathway.

Figure 1 schematically illustrates the retrosynthesis analysis of DHPV.
Figure 2a schematically illustrates a DHPV branched synthesis method according to one aspect.
Figure 2b schematically illustrates the first step of the DHPV branched synthesis method according to one aspect.
Figure 2C is a schematic diagram of the second step of the DHPV branched synthesis method according to one aspect.
FIG. 2D schematically illustrates the third step of the DHPV branched synthesis method according to one aspect.
Figure 2e schematically illustrates the fourth step of the DHPV branched synthesis method according to one aspect.
Figure 3a shows the results confirming the anti-inflammatory activity of (R)-DHPV and (S)-DHPV.
Figure 3b shows the results confirming the anti-inflammatory activity according to the concentration of (S)-DHPV.

Below, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are provided only to make the present invention easier to understand, and the content of the present invention is not limited by the following examples.

[Example]

Example 1. Retrosynthesis of DHPV

In order to evaluate the branched synthesis of DHPV and the biological properties of its isomers, first, a retrosynthesis of DHPV was planned.

Figure 1 schematically illustrates the retrosynthesis analysis of DHPV. As shown in Figure 1, (S) or (R)-DHPV can be obtained by deoxygenation and demethylation of benzyl alcohol in 2a or 2b. δ-hydroxy-γ-valerolactone (2a, 2b) is formed by Sharpless Asymmetric Dihydroxylation (SAD) and subsequent lactonization from the common intermediate alkyne (3). Alkyne (3) is formed by cross-metathesis of (4), and styrene (4) is used to lower the homo-dimerization of the final olefin. Meanwhile, the synthesis of γ,δ-unsaturated ester (3) was planned by Johnson-Claisen rearrangement instead of cross-metathesis, but meaningful results could not be obtained.

Example 2. Divergent synthesis of DHPV

2-1. Synthesis of (E)-Methyl-5-(3,4-dimethoxyphenyl)pent-4-enoate [(E)-Methyl-5-(3,4-deimethoxyphenyl)pent-4-enoate]

In 500 mL of CH ₂ Cl ₂ 967 mg, 7.54 mmol of methyl pent-4-enoate and 4-(1-propenyl)-1,2-methoxybenzene (4-(1- After stirring 2.67 g, 15.1 mmol, 2.0 equivalent of propenyl)-1,2-dimethoxybenzene), 310 mg, 0.38 mmol, 0.05 equivalent of Grubbs second generation catalyst was added and heated to reflux for 6 hours. Thereafter, the reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. 30 mL of EtOAc/n-hexane (1:10) was added, and the solid was filtered to obtain 1.76 g of styrene dimer. After purifying the filtrate by silica gel column chromatography, 160 mg of ethyl pent-4-enoate was recovered as (E)-methyl-5-(3,4-dimethoxyphenyl)pent-4, a colorless oil. -enoate [(E)-Methyl-5-(3,4-deimethoxyphenyl)pent-4-enoate] was obtained (yield 59%). Unless otherwise stated, all reactions were performed under nitrogen (N ₂ ) atmosphere under anhydrous conditions.

2-2. (S)-5((S)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one[(S)-5-((S)-(3, Synthesis of 4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one]

1.47 g, 1.40 g/mmol of AD-mix-α and 120.9 mg, 1.27 mmol, 1.21 equivalents of methane sulfonamide were stirred in 30 mL of t-BuOH/H ₂ O (1:1). Afterwards, 280 mg, 1.05 mmol of (E)-methyl-5-(3,4-dimethoxyphenyl)pent-4-enoate obtained in Example 2-1 was dissolved in 10 mL of t-BuOH, and then It was added to the stirred solution and stirred at 0°C for 72 hours. Afterwards, 100 mL of EtOAc was added to the reaction mixture and the organic layer was separated. The aqueous layer was extracted twice with 50 mL of EtOAc, combined with the organic layer, and washed twice with 10 mL of H ₂ O. The organic layer was dried with MgSO ₄ and the solvent was removed under reduced pressure. The lactone was purified by silica gel column chromatography (EtOAc/n-hexane, 2:1) to produce (S)-5((S)-(3,4-dimethoxyphenyl)(hydroxy)methyl)di as a white solid. 164 mg, 0.65 mmol of hydrofuran-2(3H)-one was obtained (yield 62%).

2-3. (R)-5((R)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one[(R)-5-((R)-(3, Synthesis of 4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one]

(R)-5((R)-(3,4-dimethoxyphenyl)(hydroxy)methyl)di in the same manner as Example 2-2 except that AD-mix-β was used. Hydrofuran-2(3H)-one was obtained (yield 55%).

2-4. Synthesis of (S)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one [(S)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one]

(S)-5((S)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2 obtained in Example 2-2 above in 20 mL of MeOH and nitrogen (N ₂ ) atmosphere. After stirring 150 mg and 0.603 mmol of (3H)-one, 20 mg of Pd(OH) ₂ was added. Thereafter, nitrogen gas was removed under reduced pressure and the reaction mixture was charged with a hydrogen gas balloon. After 6 hours, the reaction mixture was filtered through a Celite pad and the solution was removed under reduced pressure. The raw material was purified by silica gel column chromatography (EtOAc/n-hexane, 1:1), and (S)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-, a colorless oil, was obtained. 102 mg, 0.43 mmol of onion was obtained (yield 72%).

2-5. Synthesis of (R)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one [(R)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one]

(R)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one was obtained in the same manner as Example 2-4 (yield 68%).

2-6. (S)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one [(S)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one] synthesis

After stirring 50 mg, 0.212 mmol of (S)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one obtained in Example 2-4 in 10 mL of CH ₂ Cl ₂ After adding 100 L, 1.06 mmol, 5.0 equivalents of BBr ₃ at 0°C, the reaction mixture was warmed to room temperature. Afterwards, the reaction mixture was quenched with 10 mL of H ₂ O at 0°C, and the organic layer was separated. The aqueous layer was re-extracted twice with 50 mL of CH ₂ Cl ₂ , combined with the organic layer, and washed twice with 10 mL of H ₂ O. The organic layer was dried with MgSO ₄ and the solvent was removed under reduced pressure. The lactone was purified by silica gel column chromatography (CH ₂ Cl ₂ /MeOH = 10:1) to produce ((S)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H) as a white solid. )-one (26 mg, 0.127 mmol) was obtained (yield 60%).

2-7. (R)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one [(S)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one] synthesis

(R)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one was obtained in the same manner as Example 2-6 (yield 62%).

Example 3. Analysis of physicochemical properties of DHPV

The physicochemical properties of DHPV and its intermediate products synthesized in Example 2 were analyzed. Specifically, ¹ H and ¹³ C-spectra were measured with a 500 MHz JNM-ECZ 500R (JEOL, Tokyo, Japan), and chemical shifts were expressed as δ values ( ¹ H: 7.26δ, ¹³ C: 77.0δ). Infrared (IR) spectra were measured with a 1600 FT-IR spectrometer (Perkin-Elmer, Waltham, MA, USA). High-resolution mass spectrometry data were recorded by JMS-700 (JEOL, Tokyo, Japan) (FAB or ESI), and values were calculated from the molecular formula to four decimal places. Melting points (mp) were obtained using MEL-TEMP® (Cole-Parmer, Staffordshire, UK).

3-1. (E)-Ethyl-5-(3,4-dimethoxyphenyl)pent-4-enoate [(E)-Ethyl-5-(3,4-deimethoxyphenyl)pent-4-enoate]

(One) ^One H-NMR (500 MHz, CDCl3)

6.88 (d, J = 1.7 Hz, 1H), 6.85 (dd, J = 8.0, 1.7 Hz, 1H), 6.78 (d, J = 8.5 Hz, 1H), 6.35 (d, J = 15.5 Hz, 1H), 6.06 (dt, J = 16.0, 6.6 Hz, 1H), 4.13 (q, J = 7.3 Hz, 2H), 3.88 (s, 3H), 3.86 (s, 3H), 2.52-2.44 (m, 4H), and 1.24 (t, J = 7.2 Hz, 3H);

(2) ¹³ C-NMR (125 MHz, CDCl3)

173.1, 149.0, 148.5, 130.6, 130.6, 126.6, 119.1, 111.1, 108.6, 60.4, 55.9, 55.8, 34.2, 28.3, and 14.3;

(3) FT-IR (thin film, neat)

ν _{up to} 2980, 2358, 2341, 2040, 1732, 1514, 1263, 1024, and 966 cm ^-1 ;

(4) HRMS (ESI+)

265.1444 (calculated for C15H21O4 ([M + H]+): 265.1434).

3-2. (S)-5((S)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one[(S)-5-((S)-(3, 4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one]

(1) Melting point : 138-140℃;

(2) ^One H-NMR (500 MHz, CDCl3)

6.91 (d, J = 1.7 Hz, 1H), 6.89 (dd, J = 8.0, 1.7 Hz, 1H), 6.83 (d, J = 8.0 Hz, 1H), 4.63-4.59 (m, 2H), 3.87 (s) , 3H), 3.86 (s, 3H), 2.50-2.43 (m, 3H), and 2.03-1.95 (m, 2H);

(3) ¹³ C-NMR (125 MHz, CDCl3)

177.0, 149.3, 149.2, 130.8, 119.4, 111.0, 109.7, 83.5, 76.3, 56.0, 55.9, 28.5, and 24.0;

(4) FT-IR (thin film, neat)

ν _{up to} 3477, 2939, 1770, 1514, 1263, and 1095 cm ^-1 ; andHRMS

(5) (ESI+)

252.0979 (calculated for C13H16O5 ([M]+): 252.0992).

3-3. (R)-5((R)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one[(R)-5-((R)-(3, 4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one]

(1) Melting point : 129-131℃;

(2) ^One H-NMR (500 MHz, CDCl3)

6.93 (d, J = 2.3 Hz, 1H), 6.91 (dd, J = 8.6, 1.7 Hz, 1H), 6.85 (d, J = 8.6 Hz, 1H), 4.70-4.62 (m, 2H), 3.88 (s) , 3H), 3.87 (s, 3H), 2.75 (bs, 1H), 2.49-2.43 (m, 2H), and 2.03-1.99 (m, 2H);

(3) ¹³ C-NMR (125 MHz, CDCl3)

177.2, 149.2, 149.2, 130.9, 119.4, 111.0, 109.8, 83.6, 76.2, 56.0, 55.9, 28.5, and 24.0;

(4) FT-IR (thin film, neat)

ν _{up to} 3477, 2939, 1770, 1516, 1263, 1184, and 1141 cm ^-1 ;

(5) HRMS (ESI+)

252.1003 (calculated for C13H16O5 ([M]+): 252.0992).

3-4. (S)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one [(S)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one]

(One) ^One H-NMR (500 MHz, CDCl3)

6.79 (d, J = 8.1 Hz, 1H), 6.75-6.71 (m, 2H), 4.71 (quintet, J = 6.4 Hz, 1H), 3.85 (s, 3H), 3.84 (s, 3H), 2.96 (dd , J = 14.4, 5.8 Hz, 1H), 2.89 (dd, J = 14.3, 6.3 Hz, 1H), 2.50-2.22 (m, 3H), and 1.96-1.89 (m, 1H);

(2) ¹³ C-NMR (125 MHz, CDCl3)

177.2, 149.0, 148.1, 128.3, 121.6, 112.6, 111.4, 111.3, 80.9, 55.9, 55.9, 28.7, and 27.0;

(3) FT-IR (thin film, neat)

ν _{up to} 3522, 2937, 1770, 1734, 1514, 1261, 1178, 1157, 1141, and 1026 cm ^-1 ;

(4) HRMS (ESI+)

237.1129 (calculated for C13H17O4 ([M + H]+): 237.1121).

3-5. (R)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one [(R)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one]

(One) ^One H-NMR (500 MHz, CDCl3)

6.80 (d, J = 8.0 Hz, 1H), 6.76-6.73 (m, 2H), 4.71 (quintet, J = 6.9 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H), 2.97 (dd , J = 14.3, 5.8 Hz, 1H), 2.89 (dd, J = 14.4, 5.8 Hz, 1H), 2.47-2.23 (m, 3H), and 1.97-1.89 (m, 1H);

(2) ¹³ C-NMR (125 MHz, CDCl3)

177.2, 149.0, 148.2, 128.4, 121.6, 112.7, 113.4, 80.9, 56.0, 55.9, 40.9, 28.7, and 27.0;

(3) FT-IR (thin film, neat)

ν _{up to} 3477, 2939, 1772, 1516, 1263, 1184, 1143, and 1026 cm ^-1 ;

(4) HRMS (ESI+)

237.1125 (calculated for C13H17O4 ([M + H]+): 237.1121).

3-6. (S)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one [(S)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one]

(1) Melting point : 163-165℃;

(2) ^One H-NMR (500 MHz, DMSO-d6)

8.74 (bs, 2H), 6.60 (d, J = 8.0 Hz, 1H), 6.58 (d, J = 2.3 Hz, 1H), 6.44 (dd, J = 8.0, 2.3 Hz, 1H), 4.58 (quintet, J = 6.7 Hz, 1H), 2.73 (dd, J = 13.8, 6.9 Hz, 1H), 2.66 (dd, J = 13.7, 5.8 Hz, 1H), 2.41 (dd, J = 17.8, 8.6 Hz, 1H), 2.31 (ddd, J = 17.8, 9.8, 5.2 Hz, 1H), 2.14-2.07 (m, 1H), and 1.83-1.76 (m, 1H);

(3) ¹³ C-NMR (125 MHz, DMSO-d6)

177.6, 145.5, 144.4, 128.0, 120.5, 117.2, 115.9, 81.3, 40.2, 28.7, and 27.1;

(4) FT-IR (thin film, neat)

ν _{up to} 3404, 1575, 1517, 1273, and 1186 cm ^-1 ;

(5) HRMS (ESI+)

209.0811 [calculated for C11H13O4 ([M + H]+): 209.0808].

3-7. (R)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one [(S)-5-(3,4-dihydroxybenzyl)dihydrofuran-2(3H)-one]

(1) Melting point: 154~156 ℃;

(2) ^One H-NMR (500 MHz, DMSO-d6)

8.78 (bs, 2H), 6.60 (d, J = 7.5 Hz, 1H), 6.57 (d, J = 1.7 Hz, 1H), 6.44 (dd, J = 8.0, 2.3 Hz, 1H), 4.58 (quintet, J = 6.7 Hz, 1H), 2.73 (dd, J = 14.3, 6.8 Hz, 1H), 2.65 (dd, J = 14.3, 6.5 Hz, 1H), 2.41 (dd, J = 17.8, 9.2 Hz, 1H), 2.31 (ddd, J = 17.2, 9.2, 4.6 Hz, 1H), 2.13-2.08 (m, 1H), and 1.83-1.75 (m, 1H);

(3) ¹³ C-NMR (125 MHz, DMSO-d6)

177.6, 145.6, 144.4, 127.9, 120.5, 117.2, 115.9, 81.3, 40.2, 28.7, and 27.1;

(4) FT-IR (thin film, neat)

ν _{up to} 3331, 3055, 1762, 1517, 1444, 1265, 1180, and 736 cm ^-1 ;

(5) HRMS (ESI+)

209.0808 (calculated for C11H13O4 ([M + H]+): 209.0808).

Example 4. Confirmation of anti-inflammatory activity of DHPV

NF-κB is a key regulator of the immune system, and activation of NF-κB causes inflammation and cancer. Expression and activation of NF-κB are strongly induced in the inflamed intestines of patients with inflammatory bowel disease. In particular, macrophages and epidermal cells isolated from inflamed intestinal samples from patients with inflammatory bowel disease have increased levels of NF-κB subunits. Additionally, the amount of activated NF-κB is significantly related to the severity of intestinal inflammation. NF-κB is activated by various stimuli such as bacterial cell wall elements such as LPS, pro-inflammatory cytokines, viruses and DNA damaging agents. These inducers stimulate the phosphorylation of Iκβα, and translocation of NF-κB to the nucleus promotes the transcription of genes encoding inflammation-related proteins. Therefore, measuring the phosphorylation and fragmentation of IκBα is a simple but effective method to test the anti-inflammatory potential of synthetic compounds through inhibition of NF-κB in patients with inflammatory bowel disease.

Against this background, the anti-inflammatory activity of DHPV synthesized in Examples 2-6 and 2-7 was confirmed. Specifically, rat intestinal epithelial cells (ICE-6) obtained from ATCC were monolayered in DMEM medium containing 10% (v/v) fetal calf serum (ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin. It was cultured. At this time, a humid atmosphere containing 5% CO ₂ was maintained at 37°C. 25 μM each of DHPV synthesized in Examples 2-6 and 2-7 was treated and stimulated with 10 ng/ml of LPS (Sigma Chemical Co., USA) for 1 hour. Thereafter, the cell monolayer was washed twice with PBS, scraped with cold cell lysis buffer containing protease inhibitors (Roche Applied Science, Mannheim, Germany) on ice, and centrifuged to remove pellets and debris. Afterwards, the phosphorylation and degradation of IκBα, an anti-inflammatory marker, was confirmed by Western blot.

Figure 3a shows the results confirming the anti-inflammatory activity of (R)-DHPV and (S)-DHPV.

Figure 3b shows the results confirming the anti-inflammatory activity according to the concentration of (S)-DHPV.

As a result, as shown in Figure 3a, (S)-DHPV (1a) was confirmed to be more effective in inhibiting the phosphorylation and degradation of IκBα compared to (R)-DHPV (1b). That is, when comparing the anti-inflammatory effects of (S)-DHPV and (R)-DHPV in terms of phosphorylation inhibition and degradation, (S)-DHPV was more effective in suppressing NF-κB signaling than (R)-DHPV. You can see that it is effective.

Additionally, as shown in Figure 3b, (S)-DHPV (1a) was confirmed to inhibit LPS-induced phosphorylation and degradation of IκBα in a concentration-dependent manner. These results imply that (S)-DHPV can prevent or treat LPS-induced inflammation by inhibiting NF-κB through inhibition of IκBα degradation. In other words, metabolites produced in the intestines have a beneficial effect on ICE-5 cells, and the chirality of metabolites is important for anti-inflammatory activity.

The description of the present invention described above is for illustrative purposes, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. will be. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive.

Claims (13)

Synthesizing a compound represented by Formula 6 below through a cross metathesis (CM) reaction of a compound represented by Formula 4 below and a compound represented by Formula 5 below;
Synthesizing a compound represented by Formula 2 below by subjecting the compound represented by Formula 6 synthesized through the above reaction to a Sharpless Asymmetric Dihydroxylation (SAD) reaction;
Synthesizing a compound represented by Formula 7 below through hydrogenolysis of the hydroxyl group of the compound represented by Formula 2 synthesized through the above reaction; and
A method for producing DHPV (5-(3',4'-dihydroxyphenyl)-γ-valerolactone) comprising the step of deprotecting the compound represented by the following formula (7) synthesized through the above reaction:
[Formula 4]

[Formula 5]

[Formula 6]

[Formula 2]

[Formula 7]

In the above,
n is 1 to 3;
X and R ₁ are each hydrogen (H) and an alkyl group of C ₁ to C ₄ ;
R ₂ is each hydrogen (H) and an alkoxy group of C ₁ to C ₄ . The method according to claim 1, wherein the compound represented by Formula 4 is 4-(1-propenyl)-1,2-methoxybenzene (4-(1-prophenyl)-1,2-dimethoxybenzene) or 4-(1- The method is propenyl)-1,2-ethoxybenzene (4-(1-prophenyl)-1,2-dimethoxybenzene). The method according to claim 1, wherein the compound represented by Formula 4 is used in an amount of 0.1 or more per equivalent of the compound represented by Formula 5. The method according to claim 1, wherein the compound represented by Formula 6 is (E)-methyl-5-(3,4-dimethoxyphenyl)pent-4-enoate [(E)-Ethyl-5-(3,4- deimethoxyphenyl)pent-4-enoate] or (E)-Ethyl-5-(3,4-dimethoxyphenyl)pent-4-enoate[(E)-Ethyl-5-(3,4-deimethoxyphenyl)pent- 4-enoate] method. The method of claim 1, wherein the compound represented by Formula 2 is (S)-5((S)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one[( S)-5-((S)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one], (R)-5((R)-(3,4-dimethoxyphenyl )(hydroxy)methyl)dihydrofuran-2(3H)-one [(R)-5-((R)-(3,4-dimethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one] , (S)-5((S)-(3,4-diethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one[(S)-5-((S)-(3 ,4-diethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one], (R)-5((R)-(3,4-diethoxyphenyl)(hydroxy)methyl)dihydrofuran-2 (3H)-one [(R)-5-((R)-(3,4-diethoxyphenyl)(hydroxy)methyl)dihydrofuran-2(3H)-one] or a mixture thereof. The method according to claim 1, wherein the compound represented by Formula 7 is (S)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one [(S)-5-(3,4- dimethoxybenzyl)dihydrofuran-2(3H)-one], (R)-5-(3,4-dimethoxybenzyl)dihydrofuran-2(3H)-one[(R)-5-(3,4-dimethoxybenzyl )dihydrofuran-2(3H)-one], (S)-5-(3,4-diethoxybenzyl)dihydrofuran-2(3H)-one[(S)-5-(3,4-diethoxybenzyl) dihydrofuran-2(3H)-one], (R)-5-(3,4-diethoxybenzyl)dihydrofuran-2(3H)-one[(R)-5-(3,4-diethoxybenzyl)dihydrofuran -2(3H)-one] or a mixture thereof. The method of claim 1, wherein the step of deprotecting the compound represented by Formula 7 includes boron tribromide (BBr ₃ ), thiophenol (PHSH), lithium diode (LiI), methylmagnesium iodide (MeMgI), Lithium chloride (LiCl), trimethylsilyl iode (TMSI), aluminum tribromide (AlBr ₃ ), aluminum trichloride (AlCl ₃ ), boron triode (BI ₃ ), boron trifluoride (BF ₃ ), boron trichloride A method carried out using at least one catalyst selected from the group consisting of chloride (BCl ₃ ). The method according to claim 7, wherein the catalyst is used in an amount of 0.01 to 50 equivalents based on 1 equivalent of the lactone compound. delete Prevention or treatment of inflammatory bowel disease containing (S)-DHPV (5-(3',4'-dihydroxyphenyl)- γ-valerolactone), which is an enantiomer of the following formula 1, or a pharmaceutically acceptable salt thereof as an active ingredient: Pharmaceutical compositions for:
[Formula 1]
.
The method of claim 10, wherein the inflammatory bowel disease is any selected from the group consisting of enteritis, ulcerative colitis, Crohn's disease, intestinal Bechet's disease, bleeding rectal ulcer, and ileal pouchitis. A pharmaceutical composition for preventing or treating one or more inflammatory bowel diseases. The pharmaceutical composition for preventing or treating inflammatory bowel disease according to claim 10, which inhibits phosphorylation and differentiation of IκBα. A health functional food composition for preventing or improving inflammatory bowel disease containing (S)-DHPV (5-(3',4'-dihydroxyphenyl)-γ-valerolactone) or a foodologically acceptable salt thereof as an active ingredient.