WO2022270478A1 - Antiviral agent - Google Patents

Abstract

The present invention pertains to an antiviral agent that comprises a compound represented by general formula (1) or (2) [wherein R1 and R2 independently represent a hydrogen atom, an alkyl group having 1-6 carbon atoms, an acyl group having 1-6 carbon atoms, a benzyl group or a benzoyl group; p and q are independently 1 or 2; and r is an integer of 2-12], a derivative thereof, a salt thereof or a solvate of the same.

Description

antiviral agent

The present invention relates to antiviral agents that are effective as therapeutic agents against coronaviruses such as SARS-CoV-2.
This application claims priority based on Japanese Patent Application No. 2021-102280 filed on June 21, 2021, the content of which is incorporated herein.

A coronavirus is an enveloped virus whose gene is a positive single-stranded RNA. Coronaviruses that infect humans include, in addition to viruses that cause common colds, those that cause acute respiratory syndrome. For example, SARS-CoV, which is the cause of the severe acute respiratory syndrome (SARS) epidemic in 2003; There is SARS-CoV-2, which is the cause of the new coronavirus infection COVID-19. In particular, there are still few therapeutic drugs for COVID-19, and the development of more effective therapeutic drugs is required.

The entry of SARS-CoV-2 into cells is divided into a pathway (TMPRSS2 pathway) by the activity of the spike protein (S) after adsorption by the host TMPRSS2 and a pathway in which it is taken up into cells by endosomes and propagated by cathepsins in lysosomes ( Cathepsin pathway) are mainly mentioned (Non-Patent Document 1). Compounds having inhibitory activity against the TMPRSS2 pathway and the cathepsin pathway are expected as candidate compounds for therapeutic agents for SARS-CoV-2, and searches for such compounds are actively carried out worldwide. For example, remdesivir (GS-5734) has inhibitory activity on these pathways and has been approved as a therapeutic agent for COVID-19 (Non-Patent Document 2). Remdesivir is a monophosphoramidate prodrug of GS-441524, an antiviral agent consisting of a nucleic acid compound developed as a treatment for Ebola hemorrhagic fever and Marburg virus infection.

Fumihiro Taguchi, Virus, 2009, Vol. 59, No. 2, pp. 215-222. Manli Wang, et al., Cell Research, 2020, vol.30, p.269-271.

The purpose of the present invention is to provide an antiviral agent that is effective as a therapeutic drug for viral infectious diseases including COVID-19.

Edible natural products are expected to have the advantage of ensuring safety for humans and sufficiently lowering the hurdles for application to pharmaceuticals. Therefore, the present inventors screened a library of compounds derived from edible natural products, and identified compounds having inhibitory activity against the TMPRSS2 pathway and the cathepsin pathway as candidate compounds for antiviral agents against SARS-CoV-2. The present invention was completed by selecting as

That is, the present invention provides the following antiviral agents and the like.
[1] the following general formula (1) or (2)

[In the formula, R ¹ and R ² are each independently a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an acyl group having 1 to 6 carbon atoms, a benzyl group, or a benzoyl group; , each independently is 1 or 2; r is an integer from 2 to 12. ]
An antiviral agent comprising a compound represented by or a derivative thereof, a salt of these compounds, or a solvate thereof.
[2] The general formula (1) is represented by the following general formulas (1-1) to (1-4)

[In the formula, R ¹ , R ² and r are the same as defined above. ]
is either
The general formula (2) is represented by the following general formulas (2-1) to (2-4)

[In the formula, R ¹ , R ² and r are the same as defined above. ]
The antiviral agent of [1] above, which is any one of
[3] The antiviral agent of [1] or [2], which comprises a compound represented by any one of formulas (C-1) to (C-11).

[4] The antiviral agent of any one of [1] to [3], which has the ability to suppress cell invasion of coronavirus.
[5] A pharmaceutical composition comprising the antiviral agent according to any one of [1] to [4] as an active ingredient.
[6] The pharmaceutical composition according to [5] above, which is used for treating infections caused by coronaviruses.
[7] The pharmaceutical composition of [6] above, which is used for treatment of COVID-19.
[8] A food containing the antiviral agent according to any one of the above [1] to [4] as an active ingredient and ingested with the expectation of contributing to the maintenance and promotion of health.
[9] A feed containing the antiviral agent according to any one of [1] to [4] as an active ingredient.
[10] A disinfectant containing the antiviral agent according to any one of [1] to [4] as an active ingredient.

The antiviral agent according to the present invention contains, as an active ingredient, a compound with high antiviral activity against viruses such as SARS-CoV-2 that enter host cells via the TMPRSS2 pathway or the cathepsin pathway. Therefore, a pharmaceutical composition containing the antiviral agent as an active ingredient is suitable as an in vivo or ex vivo antiviral agent, and is used for the treatment or prevention of infectious diseases caused by coronaviruses such as COVID-19. It is very useful as an active ingredient of a pharmaceutical composition.

FIG. 10 shows the measurement results of the total area (μm ² ) of cells having an area of 500 μm ² or more per cell in the cell fusion inhibitory effect test of Example 4. FIG. FIG. 10 is a diagram showing the measurement results of the number of cells having an area of 500 μm ² or more per cell in the cell fusion inhibitory effect test of Example 4. FIG.

In the specification of this application, "X1 to X2 (X1 and X2 are real numbers that satisfy X1<X2)" means "X1 or more and X2 or less".

The antiviral agent according to the present invention is a compound represented by the following general formula (1) (hereinafter sometimes referred to as "compound (1)") or a compound represented by the following general formula (2) (hereinafter referred to as Sometimes referred to as "compound (2)").

In general formula (1) or (2), p and q are each independently 1 or 2.

In general formula (1) or (2), r is an integer of 2-12. In the compound represented by formula (1), r is preferably an integer of 4-12, more preferably an integer of 6-10.

In general formula (1) or (2), R ¹ and R ² each independently represent a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an acyl group having 1 to 6 carbon atoms, a benzyl group, or a benzoyl group. represents A plurality of R ¹ in one molecule may be the same group or different groups. A plurality of R ² in one molecule may be the same group or different groups.

When R ¹ and R ² are alkyl groups having 1 to 6 carbon atoms, the alkyl groups may be linear or branched. More specifically, methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, sec-butyl group, isobutyl group, tert-butyl group, n-pentyl group, sec-pentyl group, isopentyl group, An alkyl group having 1 to 6 carbon atoms such as an n-hexyl group, a sec-hexyl group and an isohexyl group is preferable, an alkyl group having 1 to 4 carbon atoms is more preferable, an alkyl group having 1 to 3 carbon atoms is even more preferable, and methyl groups, ethyl groups are even more preferred.

When R ¹ and R ² are an acyl group having 1 to 6 carbon atoms, the acyl group may be an alkyl group having 1 to 5 carbon atoms, and may be linear. It may be branched. More specifically, the alkyl group moiety is methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, sec-butyl group, isobutyl group, tert-butyl group, n-pentyl group, sec-pentyl Alkyl groups having 1 to 5 carbon atoms such as groups and isopentyl groups are preferable, alkyl groups having 1 to 4 carbon atoms are more preferable, alkyl groups having 1 to 3 carbon atoms are more preferable, and methyl and ethyl groups are even more preferable. .

More specifically, compound (1) includes compounds represented by general formulas (1-1) to (1-4). Compound (2) includes compounds represented by general formulas (2-1) to (2-4). In general formulas (1-1) to (1-4) and (2-1) to (2-4), R ¹ , R ² and r are the same as in general formula (1). As the compound (1), a compound represented by the general formula (1-1) (hereinafter sometimes referred to as "compound (1-1)") is preferable, and among the compounds (1-1), r is Compounds 6-10 are particularly preferred. As the compound (2), a compound represented by the general formula (2-1) (hereinafter sometimes referred to as “compound (2-1)”) is preferable, and among the compounds (2-1), r is Compounds 6-10 are particularly preferred.

Compound (1-1) includes compounds represented by general formulas (1-1-1) to (1-1-25). Compound (2-1) includes compounds represented by general formulas (2-1-1) to (2-1-25). In general formulas (1-1-1) to (1-1-25) and (2-1-1) to (2-1-25), r is the same as in general formula (1), and 4 to 12 is preferred, and 6-10 is more preferred. Also, R ³ is a methyl group, an ethyl group, an n-propyl group, or an isopropyl group, preferably a methyl group. A plurality of R ³ in one molecule may be the same group or different groups. Also, in the formula, Ac is an acetyl group, Bz is a benzoyl group, and Bn is a benzyl group.

As compound (1) or compound (2), compounds represented by the following formulas (C-1) to (C-11) are particularly preferable.

Compound (C-1) is Malabaricone C, a component of the spices nutmeg and mace, and is guaranteed to be safe for humans to ingest. As the compound (C-1), those extracted and purified from nutmeg or mace can be used, and chemically synthesized products can also be used. Compounds (C-2) to (C-9) can be synthesized by appropriately modifying the hydroxy group of compound (C-1) or by reducing the ketone group to a hydroxy group. Compounds (C-10) to (C-11) can be synthesized, for example, according to the synthetic routes shown in Examples described later.

In the antiviral agent according to the present invention, compound (1) and compound (2) may form a salt, and the acid or base that forms the salt includes mineral acids such as hydrochloric acid and sulfuric acid; Acids, organic acids such as citric acid; alkali metals such as sodium and potassium; alkaline earth metals such as calcium and magnesium; Compound (1) and compound (2) may also be in the form of solvates such as hydrates.

Derivatives of compound (1) and compound (2) may be used as antiviral agents according to the present invention. Derivatives of compound (1) or compound (2) are preferably derivatives from which compound (1) or compound (2) is produced by enzymatic treatment or the like in vivo. In the present invention, the derivative of compound (1) or compound (2) that can be used as an antiviral agent is obtained, for example, by prodrug conversion, which is carried out for pharmaceuticals containing a low-molecular-weight compound as an active ingredient. Derivatives are more preferred. Specifically, in general formula (1) or general formula (2), a monophosphoramidite derivative in which the hydroxy group is substituted with a monophosphoramidite group, and a monophosphorothioate derivative in which the hydroxy group is substituted with a monophosphorothioate group etc. Moreover, the derivative of compound (1) and the derivative of compound (2) may form a salt or may be in the form of a solvate such as a hydrate. As the salt, those listed above can be used.

Compound (1), derivative of compound (1), compound (2), derivative of compound (2), salts of these compounds, or solvates thereof (hereinafter, collectively referred to as "according to the present invention compound") has the ability to suppress entry into host cells via the TMPRSS2 pathway or the cathepsin pathway. Due to this ability to suppress cell entry, the compounds of the present invention have antiviral activity against viruses, particularly viruses such as SARS-CoV-2 that enter host cells via the TMPRSS2 pathway or the cathepsin pathway. , is useful as an active ingredient of a pharmaceutical composition used for treatment or prevention of viral infections.

The compound according to the present invention is particularly useful as an active ingredient of pharmaceutical compositions used for treating or preventing coronavirus infectious diseases including COVID-19. Since the compound according to the present invention is a low-molecular-weight compound, there is no problem such as immunogenicity. In addition, since the route of administration is not so limited, it is particularly useful as an active ingredient of medicines for mammals including humans.

When one or more compounds according to the present invention are contained in a pharmaceutical composition, they can be mixed with a pharmaceutically acceptable carrier, if necessary, and various dosage forms can be adopted depending on the purpose of prevention or treatment. is. Examples of such forms include oral agents, injections, sprays, suppositories, ointments, patches, etc., but oral agents are preferred. Each of these dosage forms can be produced by a conventional formulation method known to those skilled in the art. 

Pharmaceutically acceptable carriers include excipients, binders, disintegrants, lubricants, and coloring agents in solid preparations; solvents, solubilizers, suspending agents, tonicity agents, buffers in liquid preparations and analgesics, etc. are used. Formulation additives such as preservatives, antioxidants, colorants, sweeteners, stabilizers, etc. may also be used as necessary.

When preparing an oral solid preparation, excipients are added to the compounds of the present invention, and if necessary, binders, disintegrants, lubricants, coloring agents, flavoring agents, etc. are added, and the Tablets, coated tablets, granules, powders, capsules, etc. can be produced by

When preparing oral liquid preparations, flavoring agents, buffering agents, stabilizing agents, flavoring agents, etc. are added to the compounds of the present invention, and oral liquid preparations, syrups, elixirs, etc. are produced by conventional methods. can be done.

When preparing an injection, the compound according to the present invention is added with a pH adjusting agent, a buffering agent, a stabilizer, a tonicity agent, a local anesthetic, etc., and injected subcutaneously, intramuscularly or intravenously by a conventional method. agent can be manufactured.
Alternatively, a pH adjuster, a buffering agent, a stabilizer, a tonicity agent, a local anesthetic, etc. are added to the compound according to the present invention to prepare a liquid preparation, which is filled in a nebulizer to prepare a nebulizer. can do.

Suppositories can be prepared by conventional methods after adding pharmaceutical carriers known in the art such as polyethylene glycol, lanolin, cocoa butter, fatty acid triglycerides, etc. to the compound of the present invention.
When preparing an ointment, bases, stabilizers, wetting agents, preservatives and the like that are commonly used for the compounds of the present invention are blended, if necessary, and mixed and formulated by conventional methods.
When preparing a patch, the above-mentioned ointments, creams, gels, pastes, etc. may be applied to a conventional support by a conventional method.

The content of the compound according to the present invention in each of the above formulations varies depending on the patient's condition, dosage form, etc., but is generally about 0.001 to 1000 mg for oral formulations and about 0.001 to 500 mg for injections. , about 0.01 to 1000 mg for suppositories.

The daily dose of these formulations varies depending on the patient's symptoms, body weight, age, sex, etc., and cannot be determined indiscriminately. 0.01 to 1000 mg is preferable, and it is preferable to administer this amount once a day or in 2 to 3 divided doses.

The animal to which the antiviral agent and pharmaceutical composition containing the compound of the present invention as an active ingredient is administered is not particularly limited, and may be a human or a non-human animal. Non-human animals include mammals such as cows, pigs, horses, sheep, goats, monkeys, dogs, cats, rabbits, mice, rats, hamsters and guinea pigs, and birds such as chickens, quails and ducks.

The compound according to the present invention is preferably added not only to pharmaceutical compositions but also to foods. A preventive effect against viral infectious diseases such as COVID-19 can be expected by ingesting food containing the compound of the present invention. Examples of such foods include health foods, functional foods, supplements, and other foods that are ingested with the expectation of contributing to the maintenance and promotion of health. Foods containing the compound of the present invention include, for example, forms in which the compound of the present invention is inoculated in the form of tablets, liquids, capsules, and the like. In addition, the compound according to the present invention can also be in the form of powder, granules, or liquid (solution, suspension) and added to other foods for ingestion, such as so-called seasonings.

The compound according to the present invention may be contained in foods for animals other than humans. Animal food includes, for example, livestock feed and pet food (pet food). When it is contained in feed, it can be produced by a conventional method, except that the compound according to the present invention is blended together with other raw materials. It is also preferable to make the compound of the present invention into powder, granules, tablets, or liquid (solution, suspension) as a feed additive to be mixed with feed and ingested by animals.

The compound according to the present invention can also be used as an active ingredient of a disinfectant. For example, by spraying, spraying, or applying a disinfectant containing the compound of the present invention, it is possible to disinfect or disinfect viruses present in the sprayed area. In particular, marabacholine C (compound (C-1)) is a compound that is guaranteed to be safe for humans to ingest. For this reason, sprays containing Marabacholine C as an active ingredient are safe even in places and environments where sprayed liquids can be absorbed into the body, such as dining tables, food materials, food manufacturing plants, nursery schools, schools, and hospitals. can be used for

The present invention will now be described in more detail with reference to examples, but the present invention is not limited to the following examples.

<Virus>
In subsequent experiments, SARS-CoV-2 was the WK-521 strain (Wuhan strain) (hCoV-19/Japan/TY/WK-521/2020) provided by the National Institute of Infectious Diseases, α strain (UK strain) (hCoV-19/Japan/QK002/2020), and γ strain (Brazilian strain) (hCoV-19/Japan/TY7-501/2021).

<Cultured cells and medium>
• SARS-CoV-2 persistently infected cells A TMPRSS2-expressing gene was introduced into MA104 cells, a cultured cell line derived from African green monkey kidney cells, to produce MA104T cells that constitutively express TMPRSS2. These MA104T cells were infected with SARS-CoV-2 (WK-521) and cultured while periodically exchanging the medium. After viral infection, many cells died and detached due to cytopathic effects. Cells surviving after long-term culture were collected and used as SARS-CoV-2 persistently infected cells.
293TA cells An ACE2 expression gene was introduced into a cultured cell line HEK-293T cells derived from human embryonic kidney cells to prepare 293TA cells in which ACE2 was constitutively expressed.
293TA-G cells 293TA-G cells were prepared by introducing a GFP-expressing gene into HEK-293T cells (293TA cells) in which ACE2 was expressed.
293TA-M cells 293TA-M cells were prepared by introducing an M-Cherry expression gene into 293TA cells.
• Vero E6 T cells A TMPRSS2-expressing gene was introduced into Vero E6 cells, a cultured cell line derived from African green monkey kidney cells, to prepare Vero E6 T cells in which TMPRSS2 was constantly expressed.

All of these cultured cells _were cultured in a medium (2 % FBS-containing MEM). In addition, when used in subsequent experiments, the cells were adjusted in advance to an appropriate cell concentration using a culture medium.

<CPE suppression effect confirmation test>
In subsequent experiments, unless otherwise specified, the CPE inhibitory effect test of test compounds on SARS-CoV-2-infected cells was performed as follows.

(material)
MTT solution 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide (Merck) was dissolved in PBS to 5 μg/mL, and then diluted to 0.45 μm or 0.45 μm. Filtered through a 22 μm filter.
・Cell lysate (virus inactivation solution)
Prepared by adding 50 mL Triton X, 4 mL hydrochloric acid (12 mol/L) in 500 mL isopropanol.

(Preparation of test compound)
A test compound was previously diluted with DMSO or a culture medium (MEM containing 2% FBS) to an appropriate concentration, and a 2-fold serial dilution series was prepared in a 96-well plate (50 μL/well).

(MTT assay)
Cells (2.0 to 2.5×10 ⁵ cells/mL) adjusted to an appropriate cell number were dispensed at 100 μL/well into a 96-well plate containing the test compound.
Next, 50 μL/well of a SARS-CoV-2 solution that had been previously diluted to an appropriate concentration with a culture medium (MEM containing 2% FBS) was dispensed into each well of the 96-well plate. After mixing with a plate mixer, the plate was cultured in a CO ₂ incubator for 2-3 days.

The 96-well plate cultured for 2 to 3 days was observed with the naked eye or under a microscope to confirm the morphology of cells, the presence or absence of crystals, and the like. Then, 30 μL of the MTT solution was dispensed into each well and cultured in a CO ₂ incubator for 4-6 hours. After culturing, 140 μL of the supernatant was removed from each well while taking care not to absorb the cells. After 140 μL of cell lysate (virus inactivation solution) was dispensed into each well, the plate was wrapped with plastic wrap so as not to dry and left overnight at room temperature.

Then, after mixing the 96-well plate with a plate mixer, absorbance (OD) at two wavelengths of 570 nm / 630 nm was measured with a plate reader, and the effective concentration of 50% for virus-infected cells (50% cell death for virus-infected cells Inhibitory concentration (EC ₅₀ ) and 50% damaging concentration (CC ₅₀ ) against virus-uninfected cells were calculated.

EC ₅₀ is calculated based on the following formula from two points A-High (High OD, High conc.) and B-Low (Low OD, Low conc.) sandwiching the 50% OD value on the absorbance and drug concentration curve. did. In the formula, "OD" means absorbance, and "conc." means concentration of drug (test compound).

EC50= ^10Z
Z = (50% OD - Low OD) / (High OD -Low OD) x {log (High conc.) - log (Low conc.)} + log (Low conc.)

50% OD = {OD (cell control) - OD (virus control)} x 0.5 + OD (virus control) OD (cell control): the average of ODs of cell control wells OD (virus control): the average of ODs of virus control wells

<Cell fusion inhibitory effect test>
In the subsequent experiments, unless otherwise specified, the cell fusion inhibitory effects of test compounds on SARS-CoV-2 infected cells were tested as follows.

(Preparation of test compound)
A test compound was previously diluted with dimethyl sulfoxide (DMSO) or a culture medium (MEM containing 2% FBS) to an appropriate concentration, and a 2- to 5-fold serial dilution series was prepared in a 96-well plate (50 μL/well).

(Fusion test method)
SARS-CoV-2 persistently infected cells adjusted to an appropriate cell number were dispensed into 96-well plates containing test compounds at 100 μL/well, and allowed to react at room temperature for 1 hour. After that, 293TA-G cells and 293TA-M cells were adjusted to an appropriate cell number, added to each well, mixed with a plate mixer, and cultured in a CO ₂ incubator for 24 hours.

By cell fusion of SARS-CoV-2 persistently infected cells, 293TA-G cells (green fluorescence), and 293TA-M cells (red fluorescence), giant cells are formed and cells that emit yellow fluorescence are formed. Therefore, the cell fusion inhibitory effect of the test compound was evaluated using the number of cells emitting yellow fluorescence and the total area of the cells as indicators. Fluorescent cells were measured and counted using an all-in-one fluorescence microscope BZ-X800 (manufactured by Keyence Corporation) and attached software.

[Example 1]
A derivative of malavaricon C was synthesized. Malavaricon C purchased from Carbosynth was used. In addition, a compound having a methylene chain length different from that of Marabacholine C was also synthesized.

(1) Compound (C-2): Synthesis of 4-(9-(2,6-dihydroxyphenyl)-9-hydroxynonyl)benzene-1,2-diol Malavaricon C (34 mg, 0.095 mmol) in methanol (1 mL) After adding sodium borohydride (7 mg, 0.19 mmol) to the solution at 0° C., the resulting mixture was stirred at 0° C. for 4 hours to react. The reaction was monitored by thin layer chromatography (TLC) (CHCl ₃ :MeOH=10:1 (volume ratio)). After adding distilled water (3 mL) to the resulting reaction mixture, the mixture was extracted with ethyl acetate. All organic layers were combined and washed with brine, then dried over magnesium sulfate. The solvent was removed under reduced pressure to give compound (C-2) (26 mg, yield 76%).

1H NMR (500MHz, _CD3OD ) d 6.85 (t, J= ^8.06Hz , 1H), 6.64 (d, J=8.06Hz, 1H), 6.59 (s, 1H), 6.47 (d, J=8.06Hz , 1H), 6.22-6.30 (m, 2H), 5.17-5.24 (m, 1H), 2.43 (t, J=7.57Hz, 2H), 1.75-1.86 (m, 21H), 1.63-1.75 (m, 1H) ), 1.49-1.58 (m, 2H), 1.30 (br. s., 10H).
¹³ C NMR (126MHz, CD ₃ OD) D 160.5, 158.1, 155.7, 155.7, 155.5 ,115.5, 115.5, 115.5, 115.5, 115.5, 115.1 , 29.1, 28.9, 25.2.
ESI-MS (m/z): 383.18795 [M+Na] ⁺ .

(2) Compound (C-3): Synthesis of 4-(9-(2,6-diacetoxyphenyl)-9-oxononyl)-1,2-phenylene diacetate Malavaricon C (35 mg, 0.098 mmol) in pyridine (1 mL) After adding acetic anhydride (1.5 mL) to the solution at 70° C., the resulting reaction mixture was stirred at 70° C. overnight to react. The resulting reaction mixture was then extracted with ethyl acetate. All organic layers were combined and washed with brine, then dried over magnesium sulfate. The solvent was removed under reduced pressure and the crude product was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)) to give compound (C-3) (45 mg, yield 87%). got

1H NMR (500MHz, _CDCl3 ) d 7.41 (t, J= ^8.31Hz , 1H), 7.02-7.09 (m, 4H), 6.98 (d, J=1.71Hz, 1H), 2.72 (t, J=7.33 Hz, 2H), 2.58 (t, J=7.50Hz, 2H), 2.27 (d, J=1.71Hz, 6H), 2.25 (s, 6H), 1.62-1.67 (m, 2H), 1.57-1.61 (m , 2H), 1.31 (s, 8H).
¹³ C NMR (126MHz, CDCL ₃ ) D 201.3, 168.6, 168.5, 168.4, 162.8, 161.1, 147.6, 141.8, 139.8, 128.0, 126.0, 123.1 23.6, 20.9, 20.6.
ESI-MS (m/z): 549.31779 [M+Na] ⁺ .

(3) Compound (C-4): 2-(9-(4-acetoxy-3-hydroxyphenyl)nonanoyl)-1,3-phenylene diacetate (C-4-1) and 2-(9-(3-acetoxy) Synthesis of -4-hydroxyphenyl)nonanoyl)-1,3-phenylene diacetate (C-4-2) Potassium carbonate (60.0 mg, 0.43 mmol) was added, the resulting reaction mixture was stirred for 15 minutes. The reaction mixture was then cooled to 0° C., acetic anhydride (41 μL, 0.44 mmol) was added, and the resulting reaction mixture was stirred for 1 hour to react. The resulting reaction mixture was then extracted with ethyl acetate. All organic layers were combined and washed with brine, then dried over magnesium sulfate. The solvent was removed under reduced pressure and the crude product was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)) to give compound (C-4) (18 mg, yield 26%). got

¹ H NMR (500MHz, CDCl ₃ ) d 7.42 (t, J=8.18Hz, 1H), 7.06 (d, J=8.31Hz, 2H), 6.98 (d, J=8.31Hz, 1H), 6.88-6.94 ( m, 1H), 6.82 (d, J=1.95Hz, 1H), 6.72 (dd, J=2.08, 8.18Hz, 1H), 2.69-2.75 (m, 2H), 2.50-2.56 (m, 2H), 2.33 -2.35 (m, 3H), 2.25-2.27 (m, 6H), 1.62-1.67 (m, 2H), 1.55-1.60 (m, 2H), 1.31 (m, 8H).
¹³ C NMR (126MHz, CDCL ₃ ) D 201.5, 177.7, 168.6, 168.6, 162.8, 161.1, 147.6, 146.5, 144.6, 142.0, 142.0, 136.5, 135.9, 127.9, 126.8, 122.9 117.6, 117.5, 44.0, 35.3, 34.9, 31.3, 31.0, 29.3, 29.2, 29.1, 29.0, 23.6, 21.0, 20.9.
ESI-MS (m/z): 507.23164 [M+Na] ⁺ .

(4) Compound (C-5): 2-(9-(4-(benzoyloxy)-3-hydroxyphenyl)nonanoyl)-3-hydroxyphenyl benzoate (C-5-1) and 2-(9-(3-( Synthesis of benzoyloxy)-4-hydroxyphenyl)nonanoyl)-3-hydroxyphenyl benzoate (C-5-2) To a pyridine (2 mL) solution of malabarikon C (50 mg, 0.14 mmol) was added benzoyl chloride (33 μL, 0.1 mL) at 0°C. 28 mmol) was added, and the resulting reaction mixture was stirred at 0° C. for 30 minutes to react. The reaction was monitored by TLC (hexane:ethyl acetate=3:2 (volume ratio)). The resulting reaction mixture was then extracted with ethyl acetate. All organic layers were combined and washed with brine, then dried over magnesium sulfate. The solvent was removed under reduced pressure and the crude product was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)) to give compound (C-5) (22 mg, yield 28%). got

¹ H NMR (500MHz, CDCl ₃ ) d 12.73 (br. s., 1H), 8.18-8.25 (m, 4H), 7.63-7.70 (m, 2H), 7.49-7.57 (m, 4H), 7.4C- 7.48 (m, 1H), 7.09 (d, J=8.31Hz, 1H), 6.92-7.01 (m, 2H), 6.87 (d, J=1.71Hz, 1H), 6.76 (dd, J=1.83, 8.18Hz , 1H), 6.67 (dd, J=2.81, 7.94Hz, 1H), 2.89 (dt, J=4.89, 7.45Hz, 2H), 2.49-2.56 (m, 2H), 1.56-1.64 (m, 2H), 1.48-1.56 (m, 2H), 1.03-1.23 (m, 8H).
¹³ C NMR (126MHz, CDCL ₃ ) D 206.0, 165.1, 163.8, 162.1, 161.1, 146.8, 145.0, 145.0, 134.1, 134.1, 134.0, 130.0, 130.3, 128.9, 128.0, 122.1, 122.1, 122.1 117.9, 116.5, 114.7, 114.2, 44.0, 35.3, 34.9, 31.3, 31.1, 29.2, 29.0, 28.9, 23.9.
ESI-MS (m/z): 589.26470 [M+Na] ⁺ .

(5) Compound (C-6): Synthesis of 4-(9-(2,6-bis(benzoyloxy)phenyl)-9-oxononyl)-1,2-phenylene dibenzoate After adding benzoyl chloride (65 μL, 0.56 mmol) to a solution of pyridine (1 mL) at room temperature, the resulting reaction mixture was stirred overnight at room temperature to react. The resulting reaction mixture was then extracted with ethyl acetate. All organic layers were combined and washed with brine, then dried over magnesium sulfate. The solvent was removed under reduced pressure and the crude product was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)) to give compound (C-6) (66 mg, yield 61%). got

¹ H NMR (500MHz, CDCl ₃ ) d 8.14 (dd, J=1.10, 8.18Hz, 4H), 8.03-8.08 (m, 4H), 7.6C-7.67 (m, 2H), 7.48-7.55 (m, 7H) ), 7.37 (dt, J=2.44, 7.82Hz, 4H), 7.28 (d, J=8.06Hz, 1H), 7.24 (d, J=8.31Hz, 2H), 7.17 (d, J=1.95Hz, 1H ), 7.12 (dd, J=1.83, 8.18 Hz, 1H), 2.76 (t, J=7.33Hz, 2H), 2.59 (t, J=7.57Hz, 2H), 1.50-1.58 (m, 4H), 1.16 -1.23 (m, 2H), 1.06-1.14 (m, 6H).
¹³ C NMR (126MHz, CDCL ₃ ) D 201.3, 164.5, 164.3, 162.8, 147.9, 147.9, 141.8, 141.8, 140.0, 133.5, 133.5, 130.3, 130.3, 130.1, 130.1 123.1, 120.6, 44.1, 35.4, 31.1, 29.2, 29.1, 29.0, 23.5.
ESI-MS (m/z): 797.37819 [M+Na] ⁺ .

(6) Compound (C-7): 1-(2,6-dimethoxyphenyl)-9-(3,4-dimethoxyphenyl)nonan-1-one, Compound (C-8): 1-(2,6- dimethoxyphenyl)-9-(3-hydroxy-4-methoxyphenyl)nonan-1-one (C-8-1) and 1-(2,6-dimethoxyphenyl)-9-(4-hydroxy-3-methoxyphenyl)nonan- Synthesis of 1-one (C-8-2) To a stirred solution of malabarikon C (50 mg, 0.14 mmol) in DMF (N,N-dimethylformamide) (5 mL) was added potassium carbonate (97 mg, 0.70 mmol) and iodide. After adding methyl (34.86 μL, 0.56 mmol), the resulting reaction mixture was stirred at room temperature for 24 hours to react. The resulting reaction mixture was then extracted with ethyl acetate. All organic layers were combined and washed with brine, then dried over magnesium sulfate. The solvent was removed under reduced pressure and the crude product was purified by silica gel column chromatography (ethyl acetate:chloroform:hexane=1:1:2 (volume ratio)) to give compound (C-7) (17.8 mg ) and compound (C-8) (10 mg) were obtained.

Compound (C-7):
¹ H NMR (500MHz, CDCl ₃ ) δ 7.22-7.26 (1H, m), 6.77-6.79 (1H, m), 6.70-6.71 (2H, m), 6.54 (d, J=8.6Hz, 2H), 3.87 (s, 3H), 3.85 (s, 3H), 3.77 (s, 6H), 2.73 (t, J=7.5Hz, 2H), 2.54 (t, J=10Hz, 2H), 1.66 (t, J=7.2 , 2H), 1.59 (m, 2H), 1.32 (br. s. 8H).
¹³ C NMR (125MHz, CDCL ₃ ): δ 205.5, 162.1, 161.6, 148.0, 147.0, 135.6 ,135.3, 120.7, 120.1, 111.7, 111.1, 104.0, 55.9, 55.9 , 29.4, 29.4, 29.3, 29.1, 23.5.
ESI MS (m/z): 437.24581 [M+Na] ⁺ .

Compound (C-8):
¹ H NMR (500MHz, CDCl ₃ ) δ 7.23-7.26 (m, 1H), 6.75-6.82 (m, 1H), 6.65-6.68 (m, 2H), 6.54 (d, J=8.6Hz, 2H), 3.87 (d, J=9Hz, 3H), 3.77 (s, 6H), 2.73 (t, J=7.5Hz, 2H), 2.51 (dt, J=12.2, 7.8Hz, 2H), 1.66 (td, J=7.1 , 3.2Hz, 2H), 1.56-1.58 (m, 2H), 1.31 (d, J=6.6Hz, 8H).
¹³ C NMR (125MHz, CDCL ₃ ) δ 205.5, 162.8, 162.1, 156.7, 146.3, 134.9 ,130.9, 119.6, 114.6, 114.0, 114.0, 110.0, 104.0, 55.8, 55.8, 55.8, 55.8, 55.8, 55.8, 44.8, 44.8, 31.8, 31.6, 31.6, 31.6, 31.6, 31.6, 31.6 29.4, 29.2, 29.1, 23.5.
ESI MS (m/z): 423.24524 [M+Na] ⁺ .

(7) Compound (C-9): 9-(4-(benzyloxy)-3-hydroxyphenyl)-1-(2,6-bis(benzyloxy)phenyl)nonan-1-one (C-9-1) and Synthesis of 9-(3-(benzyloxy)-4-hydroxyphenyl)-1-(2,6-bis(benzyloxy)phenyl)nonan-1-one (C-9-2) Malavaricon C (50 mg, 0.14 mmol) To a stirred solution of in DMF (N,N-dimethylformamide) (5 mL) was added sodium hydride (11.17 mg, 0.28 mmol) dissolved in oil and stirred for 1 hour, then benzyl bromide (66 μL, 0.28 mmol) was added dropwise. The resulting reaction mixture was stirred at room temperature for 24 hours to react. The resulting reaction mixture was then quenched with methanol, distilled under reduced pressure, and extracted with ethyl acetate. After all organic layers were combined and dried over magnesium sulfate, the solvent was removed under reduced pressure and the crude product was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)). to obtain compound (C-9) (4.5 mg).

Compound (C-9):
¹ H NMR (500MHz, CDCl ₃ ) δ 7.17-7.42 (m, 16H), 6.82-6.85 (m, 1H), 6.75-6.78 (m, 1H), 6.67-6.69 (m, 1H), 6.59 (d, J=8.3Hz, 1H) 5.08 (s, 2H), 5.07 (s, 4H), 2.76 (t, J=7.5Hz, 2H), 2.48 (t, J=7.5Hz, 2H), 1.61-1.64 (m , 2H), 1.51-1.54 (m, 2H), 1.21-1.25 (m, 8H).
¹³ C NMR (125MHz, CDCL ³ ) Δ 206.4, 161.1, 159.4, 157.7, 155.8, 136.6, 136.5, 130.3, 130.3, 128.7, 128.7, 128.5, 128.5, 128.5, 128.5, 127.8, 127.8, 127.8, 127.8, 127.8 127.0, 127.0, 127.0, 127.0, 127.0, 121.3, 119.6, 114.8, 114.3, 112.4, 105.8, 71.3, 71.1, 70.4, 44.8, 35.6, 29.4, 29.3, 235, 27.9, 27.9, 27.9, 27.9
ESI MS (m/z): 651.40640 [M+Na] ⁺ .

(8) Compound (C-10): 1-(2,6-dihydroxyphenyl)-11-(3,4-dihydroxyphenyl)undecan-1-one and Compound (C-11): 1-(2,6-dihydroxyphenyl Synthesis of )-7-(3,4-dihydroxyphenyl)heptan-1-one

(8-1) Compound (a): Synthesis of n-benzyloxyalcohol Alkenediol (84 mmol) and DMF (100 mL) were placed in a round-bottomed flask, mixed, and then cooled to 0°C. After sodium hydride (84 mmol) was added portionwise to the mixture, the mixture was brought to room temperature and stirred for 30 minutes. The reaction mixture was then cooled to 0° C. again before benzyl bromide (84 mmol) was added dropwise and stirred continuously at room temperature for 24 hours. After the resulting reaction mixture was quenched with citric acid, the solvent was removed under reduced pressure and extracted with diethyl ether. All organic layers were combined, washed with sodium bicarbonate, dried and evaporated to give a yellow oil. The yellow oil was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)) to give the corresponding n-benzyloxyalcohol.

8-benzyloxyoctanol (10a): yellow oil, yield 52%
¹ H NMR (500MHz, CDCl ₃ ) d 7.23-7.38 (m, 5H), 4.50 (s, 2H), 3.63 (t, J=6.6Hz, 2H), 3.47 (t, J=6.6Hz, 2H), 1.52-1.68 (m, 4H), 1.33-1.46 (m, 8H).

4-benzyloxybutanol (11a): yellow oil, yield 84%
¹ H NMR (500MHz, CDCl ₃ ) d 7.23-7.37 (m, 5H), 4.50 (s, 2H), 3.59 (t, J=6.11Hz, 2H), 3.49 (t, J=6.11Hz, 2H), 1.58-1.74 (m, 4H).

(8-2) Compound (b): Synthesis of n-benzyloxyalkyl bromide N-bromosuccinimide (67 mmol) and triphenylphosphine (67 mmol) were added to a stirred solution of n-benzyl alcohol (33.5 mmol) in THF (tetrahydrofuran) at 0°C. ) was added little by little, and the mixture was stirred at room temperature for 2 hours. Methanol (10 mL) was then added to the resulting reaction mixture, which was then concentrated to half its initial volume and diluted with hexanes. After filtering the reaction mixture to remove the precipitate, the filtrate was concentrated, and the resulting crude product was purified by silica gel column chromatography (hexane:ethyl acetate=9:1 (volume ratio)) to obtain the corresponding n-benzyloxyalkyl bromide was obtained.

8-benzyloxyoctyl bromide (10b): colorless oil, yield 95%
¹ H NMR (500MHz, CDCl ₃ ) d 7.24-7.36 (m, 5H), 4.50 (s, 2H), 3.46 (t, J=6.60Hz, 2H), 3.40 (t, J=6.84Hz, 2H), 1.84 (quin, J=6.96Hz, 2H), 1.61 (quin, J=6.84Hz, 2H), 1.27-1.46 (m, 8H).

4-benzyloxybutyl bromide (11b): colorless oil, yield 75%
¹ H NMR (500MHz, CDCl ₃ ) d 7.23-7.37 (m, 5H), 4.50 (s, 2H), 3.50 (t, J=6.23Hz, 2H), 3.43 (t, J=6.72Hz, 2H), 1.93-2.01 (m, 2H), 1.71-1.80 (m, 2H).

(8-3) Compound (c): Synthesis of n-benzyloxyalkylthriphenylphosphonium Bromide After diluting n-benzyloxyalkyl bromide (18 mmol) and triphenylphosphine (55 mmol) with dichloromethane, they were reacted by heating at 80° C. for 24 hours. . After evaporating the solvent, the resulting reaction mixture was first eluted with chloroform to remove triphenylphosphine, and then purified by silica gel column chromatography (chloroform:methanol=9:1 (volume ratio)). , to give the corresponding n-benzyloxyalkylphenylphosphonium bromide.

8-benzyloxyoctyltriphenylphosphonium Bromide (10c) : colorless gel, yield 81%.
¹ H NMR (500MHz, CDCl ₃ ) d 7.67-7.90 (m, 15H), 7.22-7.36 (m, 5H), 4.47 (s, 2H), 3.76-3.85 (m, 2H), 3.42 (t, J= 6.60Hz, 2H), 1.48-1.69 (m, 6H), 1.18-1.34 (m, 6H).
ESI-MS (m/z): 481.14 [M-Br] ⁺ .

4-benzyloxybutyltriphenylphosphonium Bromide (11c): white solid, yield 65%.
¹ H NMR (500MHz, CDCl ₃ ) d 7.62-7.86 (m, 15H), 7.19-7.33 (m, 5H), 4.46 (s, 2H), 3.70-3.82 (m, 2H), 3.60 (t, J= 5.62Hz, 2H), 1.94-2.07 (m, 2H), 1.73-1.89 (m, 2H).
ESI-MS (m/z): 425.08 [M-Br] ⁺ .

(8-4) Compound (d): Synthesis of 1-benzyloxy-n-arylalkenes Phosphonium salt (4 g, 1 eq.) and sodium hydride (5 eq.) were placed in a round-bottomed flask under nitrogen gas in dry THF (100 mL). After mixing to , the resulting mixture was stirred at 0° C. for 2 hours to form the ylide. A solution of 3,4-dimethoxybenzaldehyde (1.3 eq) in dry THF (5 mL) was then added dropwise to the reaction mixture and then stirred at room temperature for 3 hours. The reaction mixture was then poured into ice water, acidified with hydrochloric acid and extracted with chloroform. All organic layers were combined, washed with sodium bicarbonate and dried to give a yellow oil. The yellow oil was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)) to give the corresponding arylalkene benzyl ether.

1-benzyloxy-9-(3,4-dimethoxyphenyl)pent-8-ene (10d): yellow oil, yield 59%.
1H NMR (500MHz, _CDCl3 ) d ^7.20-7.37 (m, 5H), 6.78-6.92 (m, 3H), 6.33 (d, J=11.48Hz, 1H), 5.58 (d, J=11.73Hz, 1H ), 4.49 (s, 2H), 3.85-3.92 (m, 6H), 3.45 (t, J=6.60Hz, 2H), 2.33 (dd, J=1.59, 7.45Hz, 2H), 1.55-1.66 (m, 2H), 1.41-1.50 (m, 2H), 1.21-1.40 (m, 8H).
ESI-MS (m/z): 390.79 [M+Na] ⁺ .

1-benzyloxy-5-(3,4-dimethoxyphenyl)pent-4-ene (11d): yellow oil, yield 62%.
1H NMR (500MHz, _CDCl3 ) d ^7.23-7.38 (m, 5H), 6.78-6.90 (m, 3H), 6.30-6.39 (m, 1H), 6.08 (td, J=6.96, 15.63Hz, 1H) , 4.52 (s, 2H), 3.83-3.92 (m, 6H), 3.52 (td, J=6.35, 10.26Hz, 2H), 2.27-2.34 (m, 2H), 1.75-1.84 (m, 2H).
ESI-MS (m/z): 334.97 [M+Na] ⁺ .

(8-5) Compound (e): Synthesis of n-arylalkanols A methanol (25 mL) solution of arylalkene benzyl ether (2 mmol) was mixed with 5% Pd/C (0.3 g) and heated to room temperature under hydrogen gas. for 24 hours, the hydrogenation reaction was carried out. After completion of the reaction, after removing the catalyst and solvent from the resulting reaction mixture, the crude product was purified by silica gel column chromatography (hexane:EtOAc=3:1 (volume ratio)) to obtain the corresponding arylalkanol. rice field.

9-(3,4-dimethoxyphenyl)nonanol (10e): colorless oil, yield 87%.
¹ H NMR (500MHz, CDCl ₃ ) d 6.77-6.84 (m, 1H), 6.70-6.77 (m, 2H), 3.88 (s, 3H), 3.89 (s, 3H), 3.66 (t, J=6.60Hz , 2H), 2.53-2.59 (m, 2H), 1.54-1.64 (m, 4H), 1.26 - 1.41 (m, 10H).
ESI-MS (m/z): 302.93 [M+Na] ⁺ .

5-(3,4-dimethoxyphenyl)pentanol (11e): colorless oil, yield 85%.
¹ H NMR (500MHz, CDCl ₃ ) d 6.78-6.83 (m, 1H), 6.71-6.76 (m, 2H), 3.87 (s, 3H), 3.89 (s, 3H), 3.65 (t, J=6.60Hz , 2H), 2.56-2.61 (m, 2H), 1.58-1.70 (m, 4H), 1.37-1.46 (m, 2H)
ESI-MS (m/z): 246.82 [M+Na] ⁺ .

(8-6) Compound (f): Synthesis of n-arylalkanals 2-iodoxybenzoic acid (IBX) was added to a stirred solution of arylalkanol (1.5 mmol) in THF/DMSO (4:1 (volume ratio)) (25 mL). (2 equivalents) was added and stirred at 40° C. for 2 hours. The resulting reaction mixture was then filtered and the filtrate was extracted with ethyl acetate. After all organic layers were combined, dried and the solvent was evaporated to give crude product. The crude product was purified by silica gel column chromatography (hexane:ethyl acetate=3:1 (volume ratio)) to give the corresponding arylalkanal.

9-(3,4-dimethoxyphenyl)nonanal (10f): colorless oil, yield 82%.
¹ H NMR (500MHz, CDCl ₃ ) d 9.78 (t, J=1.95Hz, 1H), 6.77-6.84 (m, 1H), 6.69-6.76 (m, 2H), 3.88 (s, 3H), 3.90 (s , 3H), 2.51-2.60 (m, 2H), 2.44 (dt, J=1.95, 7.33Hz, 2H), 1.63 (td, J=7.33, 19.06Hz, 4H), 1.34 (br. s., 8H) .
ESI MS(m/z): 300.81 [M+Na] ⁺ .

5-(3,4-dimethoxyphenyl)pentanal (11f): colorless oil, yield 89%.
¹ H NMR (500MHz, CDCl ₃ ) d 9.74-9.80 (m, 1H), 6.80 (d, J=7.82Hz, 1H), 6.67-6.75 (m, 2H), 3.87 (s, 3H), 3.89 (s , 3H), 2.60 (t, J=7.08Hz, 2H), 2.42-2.50 (m, 2H), 1.62-1.75 (m, 4H).
ESI-MS (m/z): 244.72 [M+Na] ⁺ .

(8-7) Compound (g): Synthesis of 2-benzyloxy-6-hydroxyacetophenone To a stirred solution of 2,6-dihydroxyacetophenone (2 g, 13.14 mmol) in DMSO (40 mL) was added sodium hydride (529 mg, 13.14 mmol). ) was added in portions at 0° C. and stirred for 30 min before benzyl bromide (1.6 mL, 13.14 mmol) was added dropwise. The reaction mixture was then brought to room temperature and kept stirring for 1 hour. The reaction mixture was then acidified with 1N cold hydrochloric acid and then extracted with dichloromethane. The residue was then purified by silica column chromatography (hexane:EtOAc=5:1 (v/v)) to give 2-benzyloxy-6-hydroxyacetophenone (2.58 g, 81%) as a yellow solid. rice field.

¹ H NMR (500MHz, CDCl ₃ ) d 13.26 (s, 1H), 7.33-7.50 (m, 5H), 6.58-6.64 (m, 1H), 6.49 (d, J=7.82Hz, 1H), 5.16 (s , 2H), 2.64 (s, 3H).
ESI-MS (m/z): 265.09 [M+Na] ⁺ .

(8-8) Condensation of 2-benzyloxy-6-hydroxyacetophenone and n-arylalkanals To a dry THF solution of 2-benzyloxy-6-hydroxyacetophenone, LDA in THF was reacted at −78° C. for 10 minutes under a nitrogen atmosphere. A solution (18 mL, 4 eq) was added dropwise and the mixture was stirred for 2 hours to produce the enolate. A solution of arylalkanal (4.5 mmol, 1 eq) and HMPA (0.5 mL) in THF (10 mL) was added to the reaction mixture and stirred for 1 h. The reaction mixture was then slowly brought to 0° C., quenched with dilute hydrochloric acid and ice, and then extracted with chloroform. After all organic layers were combined, washed with brine and solvent was evaporated under reduced pressure to give crude product. The resulting crude product was purified by silica gel column chromatography (hexane:ethyl acetate=5:1 (volume ratio)) to give the corresponding diarylalkanone.

1-(2-Benzyloxy-6-hydroxyphenyl)-11-(3,4-dimethoxyphenyl)undecan-1-one (10h): pale yellow solid, yield 36%.
¹ H NMR (500MHz, CDCl ₃ ) d 13.07 (s, 1H), 7.36-7.47 (m, 5H), 6.79-6.83 (m, 1H), 6.71-6.77 (m, 2H), 6.63 (dd, J= 0.98, 8.31Hz, 1H), 6.49-6.53 (m, 1H), 5.09-5.15 (m, 2H), 4.06-4.12 (m, 1H), 3.88 (s, 3H), 3.90 (s, 3H), 3.21 (dd, J=2.20, 18.32Hz, 1H), 3.03 (dd, J=9.53, 18.32Hz, 1H), 2.90 (d, J=3.42Hz, 1H), 2.53-2.61 (m, 2H), 1.58- 1.66 (m, 2H), 1.15-1.39 (m, 12H).
ESI MS (m/z): 543.07 [M+Na] ⁺ .

1-(2-Benzyloxy-6-hydroxyphenyl)-7-(3,4-dimethoxyphenyl)heptan-1-one (11h): pale yellow solid, yield 37%.
¹ H NMR (500MHz, CDCl ₃ ) d 13.06 (s, 1H), 7.35-7.49 (m, 5H), 6.76-6.85 (m, 1H), 6.67-6.76 (m, 2H), 6.62 (d, J= 8.31Hz, 1H), 6.50 (d, J=8.31Hz, 1H), 5.04-5.17 (m, 2H), 4.04-4.14 (m, 1H), 3.85-3.92 (m, 6H), 3.20 (dd, J =2.20, 18.32Hz, 1H), 3.03 (dd, J=9.53, 18.32Hz, 1H), 2.86-2.95 (m, 1H), 2.47-2.56 (m, 2H), 1.47-1.59 (m, 2H), 1.21-1.43 (m, 4H).
ESI MS (m/z): 486.92 [M+Na] ⁺ .

(8-9) Aldol Dehydration Reaction To a solution of aldol (1.64 mmol) in toluene (20 mL) was added a catalytic amount of p-toluenesulfonic acid (p-TsOH), and the resulting mixture was stirred at 80° C. for 0.5 hours. The reaction was allowed to heat for ~2 hours. The reaction was occasionally monitored by TLC (hexane:ethyl acetate=3:2 (volume ratio)). After completion of the reaction, the resulting reaction mixture was poured into cold water and extracted with ether. All organic layers were combined, washed with saturated sodium bicarbonate and concentrated by evaporation of the solvent under reduced pressure to give a yellow crude product. The resulting crude product was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)) to give the corresponding diarylenone.

1-(2-Benzyloxy-6-hydroxyphenyl)-11-(3,4-dimethoxyphenyl)-2-undecen-1-one (10i): yellow solid, yield 51%.
¹ H NMR (500MHz, CDCl ₃ ) d 13.12 (s, 1H), 7.32-7.48 (m, 5H), 7.17-7.23 (m, 1H), 7.00-7.08 (m, 1H), 6.79-6.83 (m, 1H), 6.71-6.77 (m, 2H), 6.63 (dd, J=0.98, 8.31Hz, 1H), 6.50 (d, J=8.31Hz, 1H), 5.12 (s, 2H), 3.87 (s, 3H) ), 3.89 (s, 3H), 2.55-2.61 (m, 2H), 2.09 (d, J=6.35Hz, 2H), 1.62 (t, J=7.33Hz, 2H), 1.22-1.39 (m, 10H) .
ESI-MS (m/z): 525.13 [M+Na] ⁺ .

1-(2-Benzyloxy-6-hydroxyphenyl)-7-(3,4-dimethoxyphenyl)-2-hepten-1-one (11i) : yellow solid, yield 60%.
¹ H NMR (500MHz, CDCl ₃ ) d 13.06 (s, 1H), 7.33-7.49 (m, 5H), 7.17 (s, 1H), 6.95-7.08 (m, 1H), 6.80 (d, J=8.31Hz , 1H), 6.66-6.75 (m, 2H), 6.63 (d, J=8.31Hz, 1H), 6.49 (d, J=8.31Hz, 1H), 5.12 (s, 2H), 3.87 (s, 3H) , 3.88 (s, 3H), 2.51 (t, J=7.82Hz, 2H), 2.12 (q, J=7.17Hz, 2H), 1.50-1.61 (m, 2H), 1.37 (quin, J=7.45Hz, 2H).
ESI-MS (m/z): 469.02 [M+Na] ⁺ .

(8-10) Hydrogenation reaction of diarylenone Add 5% Pd/C to a solution of unsaturated diarylenone (0.2 to 0.4 g) in acetone (20 mL), and hydrogenate for 3 hours under hydrogen gas. did After removing the catalyst and solvent from the reaction mixture after the completion of the reaction, the reaction mixture was purified by silica gel column chromatography (hexane:ethyl acetate=3:1 (volume ratio)) to obtain the corresponding saturated compound.

1-(2-Benzyloxy-6-hydroxyphenyl)-11-(3,4-dimethoxyphenyl)undecan-1-one (10j): pale yellow solid, yield 71%.
¹ H NMR (500MHz, CDCl ₃ ) d 9.97 (br. s., 1H), 7.21 (t, J=8.06Hz, 1H), 6.79-6.84 (m, 1H), 6.71-6.76 (m, 2H), 6.40 (d, J=8.31Hz, 2H), 3.86 (s, 3H), 3.88 (s, 3H), 3.15 (t, J=7.57Hz, 2H), 2.53-2.59 (m, 2H), 1.67-1.76 (m, 2H), 1.56-1.64 (m, 2H), 1.24-1.41 (m, 12H).
ESI-MS (m/z): 437.02 [M+Na] ⁺ .

1-(2-Benzyloxy-6-hydroxyphenyl)-7-(3,4-dimethoxyphenyl)heptan-1-one (11j): pale yellow solid, yield 88%.
¹ H NMR (500MHz, CDCl ₃ ) d 9.34 (br. s., 1H), 7.23 (t, J=8.31Hz, 1H), 6.77-6.84 (m, 1H), 6.70-6.76 (m, 2H), 6.39 (d, J=7.82Hz, 2H), 3.87 (s, 3H), 3.89 (s, 3H), 3.12 (t, J=7.33Hz, 2H), 2.57 (t, J=7.82Hz, 2H), 1.68-1.78 (m, 2H), 1.63 (quin, J=7.45Hz, 2H), 1.36-1.46 (m, 4H).
ESI-MS (m/z): 380.93 [M+Na] ⁺ .

(8-11) Demethylation Reaction of Diarylenone To a stirred solution of diarylalkanone (7.20 mmol) in dichloromethane (10 mL) was added boron tribromide (3.57 mmol) at 0° C. and then the reaction mixture was allowed to cool to room temperature. and stirred for 30 minutes. After completion of the reaction, the resulting reaction mixture was poured into ice water and extracted with ethyl acetate. All organic layers were combined, washed with saturated sodium bicarbonate and concentrated by evaporation of the solvent under reduced pressure to give a yellow crude product. The resulting crude product was purified by silica gel column chromatography (hexane:ethyl acetate=4:1 (volume ratio)) to give the corresponding diarylalkanone.

1-(2,6-dihydroxyphenyl)-11-(3,4-dihydroxyphenyl)undecan-1-one (10) : pale yellow solid, yield 45%.
1H NMR (500MHz, _CD3OD ) d 7.17 (t, J= ^8.06Hz , 1H), 6.66 (d, J=7.82Hz, 1H), 6.61 (d, J=1.95Hz, 1H), 6.47 (dd , J=1.95, 7.82Hz, 1H), 6.34 (d, J=8.31Hz, 2H), 3.02-3.16 (m, 2H), 2.38-2.46 (m, 2H), 1.66 (t, J=7.08Hz, 2H), 1.53 (t, J=7.08Hz, 2H), 1.30-1.39 (m, 4H), 1.28 (br. s., 8H).
¹³ C NMR (125MHz, CD ₃ OD) δ 208.3, 142.0, 142.6, 135.4, 135.4, 119.4, 115.8, 110.8, 110.0, 107.0, 107.0, 44.4, 31.5, 29.4, 29.4, 29.3 .
ESI-MS (m/z): 409.06 [M+Na] ⁺ .

1-(2,6-dihydroxyphenyl)-7-(3,4-dihydroxyphenyl)heptan-1-one (11) : pale yellow solid, yield 56%.
1H NMR (500MHz, _CD3OD ) d ^7.13-7.21 (m, 1H), 6.66 (d, J=7.82Hz, 1H), 6.62 (d, J=1.95Hz, 1H), 6.48 (dd, J= 1.95, 7.82Hz, 1H), 6.34 (d, J=8.31Hz, 2H), 3.09 (t, J=7.57Hz, 2H), 2.43 (t, J=7.57Hz, 2H), 1.65 (t, J= 7.33Hz, 2H), 1.48-1.59 (m, 2H), 1.26-1.42 (m, 4H).
^13C NMR (125MHz, _CD3OD ) δ 208.3, 162.0, 144.5, 142.6, 135.5, 134.4, 119.3, 115.1, 114.8, 110.0, 107.0, 44.4, 34.8, 31.4, 29.0, 24.38,
ESI-MS (m/z): 353.21 [M+Na] ⁺ .

[Example 2]
The antiviral activity against SARS-CoV-2 of malavaricon C and its structural analogues and their derivatives was examined by MTT assay. WK-521 (Wuhan strain), α strain (British strain), and γ strain (Brazilian strain) provided by the National Institute of Infectious Diseases were used as SARS-CoV-2.

As test compounds, malavaricon C (manufactured by Carbosynth) (compound (C-1)) and compounds (C-2) to (C-11) synthesized in Example 1 (derivatives of malavaricon C (C-2) ~ (C-9) and compounds (C-10) ~ (C-11) that differ in methylene chain length from malabaricone C), and remdesivir (RD) and molnupiravir (MK-4482), which are approved as therapeutic agents for COVID19 was used. For each test compound, a 2-fold serial dilution series was prepared in advance in a culture medium.

A 2-fold serial dilution series of the test compound was prepared at 50 μL/well in a 96-well plate, and Vero E6T cells adjusted to 2.0×10 ⁵ cells/mL were dispensed at 100 μL/well. Then, 50 μL/well of SARS-CoV-2 (WK-521) solution, which had been diluted to an appropriate concentration in advance, was dispensed into each well, mixed with a plate mixer, and placed in a CO ₂ incubator for 2 to 2 hours. Cultured for 3 days.

An MTT solution was added to the cultured cells and an MTT assay was performed to determine _EC50 and _CC50 . Table 1 shows the results.

As shown in Table 1, all of the compounds (C-1) to (C-11), like remdesivir and the like, have the effect of inhibiting cell death due to SARS-CoV-2 infection, and can be used as antiviral agents. shown to be useful. In particular, compounds (C-1) to (C-5) and compound (C-11) have a large selectivity index of 5 or more, and are expected as active ingredients of COVID19 therapeutic drugs.

[Example 3]
It was investigated by MTT assay whether malavaricon C and its structural analogues and their derivatives also have antiviral activity against various mutants of SARS-CoV-2.

Specifically, as SARS-CoV-2, α strain (British strain) or γ strain (Brazilian strain) was used, and Vero E6T cells or 293TA cells were used as cells to be infected in the same manner as in Example 2. Then, the cells were infected with the virus and the MTT assay was performed. The results are shown in Tables 2 and 3. In the table, "ND" indicates no data.

As shown in Tables 2 and 3, the compounds according to the present invention also showed antiviral activity against SARS-CoV-2 other than the WK-521 strain (Wuhan strain).

[Example 4]
The ability of malavaricon C and its derivatives to suppress SARS-CoV-2 entry into host cells was examined by a cell fusion inhibitory effect test. For SARS-CoV-2, the WK-521 strain (Wuhan strain) provided by the National Institute of Infectious Diseases was used.

Compound (C-1), compound (C-3), and compound (C-5) were used as test compounds. Among these, the compound (C-1) (malavaricon C) was purchased from Carbosynth, and the others were synthesized in Example 1. For comparison, two types of neutralizing antibodies (neutralizing antibody Ab1 and neutralizing antibody Ab2) were used. These antibodies are known to inhibit adhesion of SARS-CoV-2 to host cells and suppress entry of SARS-CoV-2 into host cells.

Compound (C-1), compound (C-3), and compound (C-5) were added to 6 wells each in a 2-fold serial dilution series from 25 μM. Neutralizing antibody Ab1 and neutralizing antibody Ab2 were added to 6 wells each in a 5-fold serial dilution series from 25 μg/mL. As controls, 3 wells (CC) to which neither the virus nor the test compound was added, and 3 wells (VC) to which only the virus was added without the test compound were set.

For each well after culturing for 24 hours, cells having an area of 500 μm ² or more per cell were defined as cells in which two or more cells were fused, and the number and total area (μm ² ) of these fused cells were counted. FIG. 1 shows the measurement results of the total area (μm ² ) of cells having an area of 500 μm ² or more per cell, and FIG. 2 shows the measurement results of the number of cells. As a result, compound (C-1), compound (C-3), compound (C-5), neutralizing antibody Ab1, and neutralizing antibody Ab2 all decreased the number of fused cells in a concentration-dependent manner. , the total area of fused cells was also smaller. From these results, compound (C-1), compound (C-3), and compound (C-5) are similar to neutralizing antibody Ab1 and neutralizing antibody Ab2, cell fusion by SARS-CoV-2, That is, it was confirmed that entry of SARS-CoV-2 into host cells could be suppressed.

Claims (10)

General formula (1) or (2) below

[In the formula, R ¹ and R ² are each independently a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an acyl group having 1 to 6 carbon atoms, a benzyl group, or a benzoyl group; , each independently is 1 or 2; r is an integer from 2 to 12. ]
An antiviral agent comprising a compound represented by or a derivative thereof, a salt of these compounds, or a solvate thereof. The general formula (1) is general formulas (1-1) to (1-4)

[In the formula, R ¹ , R ² and r are the same as in the general formula (1). ]
is either
The general formula (2) is general formulas (2-1) to (2-4)

[In the formula, R ¹ , R ² and r are the same as in the general formula (1). ]
The antiviral agent according to claim 1, which is any one of The antiviral agent according to claim 1, comprising a compound represented by any one of formulas (C-1) to (C-11).

The antiviral agent according to claim 1, which has the ability to suppress cell invasion of coronavirus. A pharmaceutical composition comprising the antiviral agent according to any one of claims 1 to 4 as an active ingredient. The pharmaceutical composition according to claim 5, which is used for treating infections caused by coronavirus. The pharmaceutical composition according to claim 6, which is used for treating COVID-19. A food that contains the antiviral agent according to any one of claims 1 to 4 as an active ingredient and is ingested with the expectation that it will contribute to the maintenance and promotion of health. A feed comprising the antiviral agent according to any one of claims 1 to 4 as an active ingredient. A disinfectant containing the antiviral agent according to any one of claims 1 to 4 as an active ingredient.

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