JP6359409B2 - Funiculosin derivative having toll-like receptor 4 activation action and use thereof - Google Patents

Description

  The present invention relates to a funiculosin derivative having a toll-like receptor 4 activating effect, a method for producing the same, and use of the derivative.

  Toll-like receptors (TLRs) have been reported to be involved in innate immunity and to recognize pathogenic microorganisms. Among them, toll-like receptor 4 (TLR4) is one of the typical receptors that activates the innate immune system that plays an important role in enhancing the immunogenicity of antigens in infection protection and vaccines. . MD-2 and CD14 are essential for ligand recognition of TLR4. When lipopolysaccharide, which is a major component of the outer membrane of Gram-negative bacteria, binds to TLR4 / MD-2 complex, it is composed of NF-κB and the like Activates the signaling cascade and produces inflammatory cytokines. The production of inflammatory cytokines causes an immune response.

  Thus, since activation of TLR4 causes an immune reaction, a compound having an action of activating TLR4 can be an immunostimulant, a therapeutic drug for immunodeficiency, a drug that suppresses the growth of cancer cells, and the like. For example, Patent Document 1 discloses an immunogenic composition containing a TLR4 agonist, and LPS, LPS fragments and the like are described as the TLR4 agonist. In addition, paclitaxel has been reported as a substance having a TLR4 activation action (Non-patent Document 1).

Special table 2013-525450 gazette

Kawasaki et al. (2000) J. Biol. Chem. 275 (4): 2251-2254

  It is an object of the present invention to provide a compound, a pharmaceutical, etc. having a TLR4 activating action or immunomodulating action in a rodent animal such as a mouse or human and useful for infection protection, enhancement of antigen immunogenicity, etc. And

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that funiculosin derivatives, for example, a compound represented by the following formula (I) and a compound represented by the following formula (II) are activated by mouse TLR4. The compound represented by the following formula (I), the following formula (III) and the compound represented by the following formula (IV) have been found to exhibit human TLR4 activation action and have an immunomodulatory action. A compound having a TLR4 activation action is useful for the prevention or treatment of a disease involving TLR4 (for example, a disease caused by a decrease or inactivation of TLR4). Moreover, the compound which has TLR4 activation action, such as the compound represented by the following formula (I), the compound represented by the following formula (II), the compound represented by the following formula (III), and the following formula (IV), For example, it is useful as an active ingredient such as an immunostimulant, an infection prevention preventive or therapeutic agent, an antigen immunogenicity enhancer in a vaccine, and the like. The present inventors have further studied based on the above findings and completed the present invention. That is, the present invention is as follows.

[1] The following general formula (1)

(In the above formula, X represents CH ₂ , CH (OH) or an oxygen atom,
n represents an integer of 0 or 1,
R ¹ represents a hydrogen atom or a linear or branched alkyl having 1 to 6 carbon atoms,
R ² represents a hydrogen atom, —CH ₂ CH ₂ OH, or a linear or branched alkyl group having 1 to 6 carbon atoms having an acidic group,
R ³ is the following formula (2)

(In the above formula, the wavy line indicates the bond to the 3- position of the pyridone ring) or the following formula (3)

(Wherein the wavy line represents a bond to the 3- position of the pyridone ring),
R ⁴ , R ⁵ , R ⁶ , and R ⁷ are the same or different and may have a hydroxyl group, an optionally substituted —O-acyl group having 1 to 6 carbon atoms, or a substituent. Represents a good linear or branched alkoxy group having 1 to 6 carbon atoms, or —O-benzyl group, or R ⁴ and R ⁵ , R ⁵ and R ⁶ , or R ⁶ and R ⁷ together Or a pharmaceutically acceptable salt thereof, which may form a cyclic acetal group.
[2] The compound represented by the general formula (1) is represented by the following formula (I)

Or the following formula (II)

Or the following formula (III)

Or the following formula (IV)

The compound according to [1], which is a compound represented by the formula:
[3] A pharmaceutical composition comprising the compound according to [2] or a pharmaceutically acceptable salt thereof.
[4] A toll-like receptor 4 activator comprising the compound according to [2] or a pharmaceutically acceptable salt thereof as an active ingredient.
[5] An immunostimulator comprising the compound according to [2] or a pharmaceutically acceptable salt thereof as an active ingredient.
[6] A preventive or therapeutic agent for a disease associated with Toll-like receptor 4, characterized by comprising the compound according to [2] above or a pharmaceutically acceptable salt thereof as an active ingredient.
[7] A preventive or therapeutic agent for infection prevention comprising the compound according to [2] or a pharmaceutically acceptable salt thereof as an active ingredient.
[8] The pharmaceutical composition according to the above [3], which is a composition for enhancing the immunogenicity of an antigen.

  According to the present invention, a funiculosin derivative having a mouse or human TLR4 activation action is provided. The present invention also provides a funiculosin derivative having an immunomodulatory action. The compound of the present invention is useful as an active ingredient such as an immunostimulant, a preventive or therapeutic agent for infection prevention, and an immunogenicity enhancer for an antigen. In addition, the compound of the present invention, for example, the compound represented by the above formula (I), the compound represented by the above formula (II), the compound represented by the above formula (III), and the above formula (IV). Since the compound has low toxicity, it has low side effects and is highly safe.

FIG. 1 is a diagram showing the results (FACS histogram) of the frequency of Ba / F3 cells expressing GFP analyzed by flow cytometry. 1A to 1D show the results of analysis using mouse TLR4 expressing cells, and FIGS. 1E to 1H show the results of using human TLR4 expressing cells. FIG. 1A shows Ba / F3 cells added with compound (I) at a final concentration of 10 μg / mL, FIG. 1B shows Ba / F3 cells added with compound (II) at a final concentration of 50 μg / mL, and FIG. Ba / F3 cells to which II) was added at a final concentration of 100 μg / mL, FIG. 1D is Ba / F3 cells to which a positive control (Lipid A) was added. FIG. 1E shows Ba / F3 cells added with compound (I) at a final concentration of 50 μg / mL, FIG. 1F shows Ba / F3 cells added with compound (III) at a final concentration of 50 μg / mL, and FIG. Ba / F3 cells to which IV) was added at a final concentration of 50 μg / mL, FIG. 1H is Ba / F3 cells to which a positive control (Lipid A) was added. The left peak (white) in FIGS. 1A to 1H is a DMSO (control) peak. FIG. 2 is a view showing the results of a toxicity test of a compound against Ba / F3 cells, and FIGS. 2A to 2E are analysis results using mouse TLR4-expressing cells, and FIGS. 2F to 2I are analysis results using human TLR4-expressing cells. FIG. 2A is a FACS development diagram of cells added with compound (I) at a final concentration of 10 μg / mL, and FIG. 2B is a FACS development of cells added with compound (II) at a final concentration of 100 μg / mL. Fig. 2C is a FACS development view of cells to which compound 3 obtained in step 2 of Example 2 was added to a final concentration of 50 µg / mL. Fig. 2D is a FACS of cells to which 3 ng / mL lipid A was added. FIG. FIG. 2E is a control (when DMSO is added). FIG. 2F is a FACS development diagram of cells added with compound (I) at a final concentration of 50 μg / mL, and FIG. 2G is a FACS development of cells added with compound (III) at a final concentration of 50 μg / mL. FIG. 2H is a FACS development view of cells to which compound (IV) was added to a final concentration of 50 μg / mL, and FIG. 2I is a FACS development view of cells to which 3 ng / mL lipid A was added.

The present invention relates to a compound represented by the above general formula (1) (hereinafter also referred to as compound (1) of the present invention) and a pharmaceutically acceptable salt thereof. The compound (1) of the present invention preferably contains funiculosin (in the general formula (1), n is 0, R ¹ is a methyl group, R ² is a hydrogen atom, and R ³ is represented by the following formula ( 3-1)

In which R ⁴ , R ⁵ , R ⁶ , and R ⁷ are a hydroxyl group).
Since the compound (1) of the present invention may exist in the form of hydrate and / or solvate, these hydrate and / or solvate are also encompassed in the compound of the present invention.

In the general formula (1), X represents CH ₂ , CH (OH) or an oxygen atom (O). X is preferably CH (OH) or an oxygen atom (O).
n represents an integer of 0 or 1. n is preferably 0.

R ¹ represents a hydrogen atom or a linear or branched alkyl having 1 to 6 carbon atoms. The linear or branched alkyl having 1 to 6 carbon atoms is, for example, methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, sec-butyl group, tert-butyl group, pentyl group, isopentyl group. , Neopentyl group, tert-pentyl group, 2-methylbutyl group, 1-ethylpropyl group, hexyl group, isohexyl group, 4-methylpentyl group, 3-methylpentyl group, 2-methylpentyl group, 1-methylpentyl group, 3,3-dimethylbutyl group, 2,2-dimethylbutyl group, 1,1-dimethylbutyl group, 1,2-dimethylbutyl group, 1,3-dimethylbutyl group, 2,3-dimethylbutyl group, 1- An ethylbutyl group or a 2-ethylbutyl group is preferable, and a linear or branched alkyl group having 1 to 4 carbon atoms is preferable. Among these, a methyl group, an ethyl group, or an isopropyl group is preferable, a methyl group or an ethyl group is more preferable, and a methyl group is more preferable.

R ² represents a hydrogen atom, —CH ₂ CH ₂ OH, or a linear or branched alkyl group having 1 to 6 carbon atoms having an acidic group. The linear or branched alkyl group having 1 to 6 carbon atoms is the same as described above, and is preferably a methyl group or an ethyl group. Examples of the acidic group include —S (O) OH ₂ group, —OPO (OH) ₂ group, —OP (O) (OBn) ₂ group (Bn represents benzyl group), —COOH, and the like. As the linear or branched alkyl group having 1 to 6 carbon atoms having an acidic group, —CH ₂ CH ₂ OPO (OH) ₂ , —CH ₂ COOH and the like are preferable. R ² is preferably a hydrogen atom, —CH ₂ COOH or —CH ₂ CH ₂ OP (O) (OH) ₂ . The acidic group may be an ion.

R ³ is the following formula (2)

(In the above formula, the wavy line indicates the bond to the 3- position of the pyridone ring) or the following formula (3)

(Wherein the wavy line represents a bond to the 3- position of the pyridone ring).
The group having the structure represented by the above formula (2) is preferably the following formula (2-1):

(In the above formula, the wavy line represents a bond to the 3- position of the pyridone ring).

  The group having the structure represented by the above formula (3) is preferably the following formula (3-1):

(In the above formula, the wavy line represents a bond to the 3- position of the pyridone ring).

R ⁴ , R ⁵ , R ⁶ , and R ⁷ are the same or different and may have a hydroxyl group, an optionally substituted —O-acyl group having 1 to 6 carbon atoms, or a substituent. Represents a good linear or branched alkoxy group having 1 to 6 carbon atoms, or —O-benzyl group, or R ⁴ and R ⁵ , R ⁵ and R ⁶ , or R ⁶ and R ⁷ together A cyclic acetal group may be formed.

  Examples of the -O-acyl group having 1 to 6 carbon atoms include -O-acetyl group, -O-propionyl group, -O-butynyl group, -O-valeryl group, -O-caproyl group and the like. Preferably, it is -O-acetyl group.

  The linear or branched alkoxy group having 1 to 6 carbon atoms is a formula: —O— (a linear or branched alkyl group having 1 to 6 carbon atoms), and the alkyl group has 1 to 1 carbon atoms described above. And the same as the 6 straight-chain or branched alkyl groups. Examples of the linear or branched alkoxy group having 1 to 6 carbon atoms include methoxy, ethoxy, n-propoxy, isopropyloxy, n-butoxy, isobutyloxy, sec-butyloxy, tert-butyloxy (tert-butoxy), pentyl Examples include oxy, tert-pentyloxy, hexyloxy and the like. Preferred is an alkoxy group having 1 to 4 carbon atoms, that is, an alkoxy group represented by the formula: —O— (a linear or branched alkyl group having 1 to 4 carbon atoms), and more preferred is methoxy, ethoxy, n -Propoxy or isopropyloxy.

The substituent in the C1-C6 -O-acyl group which may have a substituent and the C1-C6 linear or branched alkoxy group which may have a substituent is, for example, , Hydroxyl group, benzyl group and the like.
The substituent is preferably a hydroxyl group or the like. The -O-acyl group having 1 to 6 carbon atoms and the linear or branched alkoxy group having 1 to 6 carbon atoms may have the same or different two or more substituents.

For example, the linear or branched alkoxy group having 1 to 6 carbon atoms which may have a substituent is preferably represented by the formula: —O— (linear or branched alkyl having 1 to 6 hydroxy carbon atoms). Group).
The term “linear or branched alkyl group having 1 to 6 hydroxy carbon atoms” means that the above-mentioned “straight or branched alkyl group having 1 to 6 carbon atoms” is mono- or disubstituted, preferably mono-hydroxy. Substituted hydroxy C ₁ -C ₆ alkyl groups, such as hydroxymethyl, 2-hydroxyethyl, 1-hydroxy-1-methylethyl, 1,2-dihydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl, 5 -Hydroxypentyl, 6-hydroxyhexyl and the like. A hydroxy C ₁ -C ₄ alkyl group is preferable, and hydroxymethyl is more preferable.

The cyclic acetal formed by R ⁴ and R ⁵ , R ⁵ and R ⁶ , or R ⁶ and R ⁷ together may have a substituent. Examples of the substituent include a linear or branched alkyl group having 1 to 6 carbon atoms and a benzyl group.
Examples of the cyclic acetal include isopropylidene, benzylidene, p-methoxybenzylidene and the like. Preferably, it is isopropylidene. In the compound of the present invention, when R ⁴ and R ⁵ , R ⁵ and R ⁶ , or R ⁶ and R ⁷ are combined to form a cyclic acetal, preferably R ⁴ and R ⁵ are combined. And R ⁶ and R ⁷ together form a cyclic acetal (preferably isopropylidene).

R ⁴ , R ⁵ , R ⁶ and R ⁷ are preferably a hydroxyl group. In yet another preferred embodiment, R ⁴ , R ⁵ , R ⁶ , and R ⁷ are each a cyclic acetal (preferably R ⁴ and R ⁵ together and R ⁶ and R ⁷ together). Form isopropylidene).

  As the compound (1) of the present invention, for example, the following formula (I)

A compound represented by the following formula (II):

A compound represented by the following formula (III)

A compound represented by formula (IV):

The compound etc. which are represented by these are preferable. In the above formula (II), R ⁴ and R ⁵ are joined together, and R ⁶ and R ⁷ are joined together to form a cyclic acetal (isopropylidene).
In the present specification, the compound represented by the formula (I) is also referred to as a compound (I). The compound represented by the above formula (II) is also referred to as compound (II). The compound represented by the above formula (III) is also referred to as compound (III). The compound represented by the above formula (IV) is also referred to as compound (IV).

  Examples of pharmaceutically acceptable salts include inorganic acid salts such as hydrochloride, hydrobromide, nitrate, sulfate and phosphate; benzenesulfonate, benzoate, citrate, fumarate, glucone Acid addition salts such as organic acid salts such as acid salts, lactate salts, maleate salts, malate salts, oxalate salts, methanesulfonate salts, and tartrate salts; alkali metal salts such as sodium salts and potassium salts; magnesium salts, Alkaline earth metal salts such as calcium salts; Metal salts such as aluminum salts and zinc salts; Ammonium salts such as ammonium and tetramethylammonium; Organic amine addition salts such as morpholine addition salts and piperidine addition salts; Glycine addition salts and phenylalanine additions And amino acid addition salts such as salts, lysine addition salts, aspartic acid addition salts, glutamic acid addition salts, and the like.

  Compound (1) of the present invention, preferably Compound (I), Compound (II), Compound (III), Compound (IV), and pharmaceutically acceptable salts thereof, have Toll-like receptor 4 (TLR4) activity. It has a chemical action. In addition, the compound (1) of the present invention, for example, the compound (I), the compound (II), the compound (III), the compound (IV) and pharmaceutically acceptable salts thereof have an immunomodulatory action.

  Compound (1) of the present invention, preferably Compound (I), Compound (II), Compound (III), Compound (IV) and pharmaceutically acceptable salts thereof, are Toll-like receptor 4 activators, It is suitably used as an active ingredient such as an immunostimulator. In addition, since the compound of the present invention has Toll-like receptor 4 activating action, it prevents or treats diseases associated with Toll-like receptor 4 (for example, diseases caused by decrease or inactivation of TLR4). It is suitably used for this purpose. The disease associated with Toll-like receptor 4 is not particularly limited, and examples thereof include a state where immunity of a living body is lowered due to infection or the like. In addition, the compounds of the present invention (preferably the above-mentioned compound (I), compound (II), compound (III), compound (IV)) and pharmaceutically acceptable salts thereof prevent infection by microorganisms, or It can be used as an active ingredient of an infection prevention or treatment agent for treating infectious diseases caused by microorganisms. Further, the compound (1) of the present invention, preferably the compound (I), the compound (II), the compound (III) or the compound (IV) and their pharmaceutically acceptable salts are used as an immunogenicity of an antigen in a vaccine or the like. Is preferably used as an adjuvant. The compound (1) of the present invention, preferably the compound (I), the compound (II), the compound (III) or the compound (IV) and pharmaceutically acceptable salts thereof are also used as an active ingredient of an antiallergic agent. obtain.

  The present invention also encompasses a pharmaceutical composition containing compound (1) of the present invention (preferably compound (I), compound (II), compound (III) or compound (IV)), or a pharmaceutically acceptable salt thereof. To do. The pharmaceutical composition of the present invention may contain two or more of the compounds of the present invention and pharmaceutically acceptable salts thereof.

  The pharmaceutical composition of the present invention comprises the compound of the present invention (preferably Compound (I), Compound (II), Compound (III) or Compound (IV)), which is an active ingredient (active ingredient), or a pharmaceutically acceptable product thereof. The salt is appropriately mixed with one or more pharmaceutically acceptable carriers and the like, and is produced by any method well known in the technical field of pharmaceutics. The content of the compound (1) of the present invention and the pharmaceutically acceptable salt thereof in the pharmaceutical composition varies depending on the dosage form, dosage, etc., but is, for example, about 0.1 to 100% by weight of the whole composition. is there. When the pharmaceutical composition contains two or more of the compound (1) of the present invention and pharmaceutically acceptable salts thereof, the total of these is set to the above ratio.

  The pharmaceutical composition of the present invention comprises tablets, capsules, granules, powders, troches, syrups, emulsions, suspensions and other oral preparations; external preparations, suppositories, injections, eye drops, nasal preparations, It can be made into dosage forms, such as parenteral agents, such as a pulmonary agent.

  As the pharmaceutically acceptable carrier, various conventional organic or inorganic carrier materials used in the production of pharmaceutical preparations can be used. For example, in the case of a solid preparation, excipients, disintegrants, binders, fluidizing agents, lubricants, etc. may be mentioned, and in the case of liquid preparations, solvents, solubilizers, suspending agents, isotonicity Agents, buffering agents, soothing agents and the like. Furthermore, additives such as preservatives, antioxidants, colorants, sweeteners and the like can be used as necessary.

Examples of the excipient include lactose, mannitol, glucose, microcrystalline cellulose, starch and the like.
Examples of the disintegrant include carmellose, carmellose calcium, carmellose sodium, carboxymethyl starch sodium, croscarmellose sodium, crospovidone, hydroxypropylmethylcellulose, and crystalline cellulose.
Examples of the binder include hydroxypropyl cellulose, polyvinyl pyrrolidone, magnesium aluminate metasilicate, starch and the like.

Examples of the fluidizing agent include light anhydrous silicic acid and magnesium stearate.
Examples of the lubricant include magnesium stearate, calcium stearate, talc, colloidal silica and the like.

Examples of the solvent include purified water, ethanol, propylene glycol, macrogol, sesame oil, corn oil, olive oil and the like.
Examples of the solubilizer include propylene glycol, D-mannitol, benzyl benzoate, ethanol, triethanolamine, sodium carbonate, sodium citrate and the like.
Examples of the suspending agent include benzalkonium chloride, carmellose, hydroxypropylcellulose, propylene glycol, povidone, methylcellulose, glyceryl monostearate and the like.

Examples of the isotonic agent include glucose, D-sorbitol, sodium chloride, D-mannitol and the like.
Examples of the buffer include sodium hydrogen phosphate, sodium acetate, sodium carbonate, sodium citrate and the like.
Examples of soothing agents include benzyl alcohol.
Examples of the preservative include ethyl paraoxybenzoate, chlorobutanol, benzyl alcohol, sodium dehydroacetate, sorbic acid and the like.

Examples of the antioxidant include sodium sulfite and ascorbic acid.
Examples of the colorant include food coloring (food red No. 2 or 3, food yellow No. 4 or 5, etc.), β-carotene and the like.
Examples of the sweetening agent include saccharin sodium, dipotassium glycyrrhizinate, aspartame and the like.

  The pharmaceutical composition of the present invention is suitably used as a toll-like receptor 4 activator, immunostimulatory agent, antiallergic agent, preventive or therapeutic agent for diseases associated with toll-like receptor 4, preventive or therapeutic agent for infection prevention, etc. Used. In addition, since the pharmaceutical composition of the present invention is suitably used for enhancing the immunogenicity of an antigen, it is also suitably used as an immunogenicity enhancing agent for an antigen in, for example, a vaccine.

  “Treatment” shall mean alleviating or curing and alleviating a symptom or disease and / or its attendant symptoms. “Prevention” means delaying or preventing the onset of a symptom or illness and its attendant symptoms, preventing a subject from acquiring a symptom or illness, or the risk of a subject acquiring a symptom or illness. It means to reduce.

  Examples of diseases associated with Toll-like receptor 4 include a state in which immunity of a living body is lowered due to immunodeficiency, infection with microorganisms, and the like. For example, when the immunity of the living body is lowered due to infection or the like, the homeostasis of the living body can be maintained by stimulating the immunity by administering the pharmaceutical composition of the present invention.

The pharmaceutical composition of the present invention is useful, for example, for protecting microorganisms from infection. Although the kind of microorganism is not specifically limited, Usually, bacteria, fungi, parasites or protozoa, virus, mycoplasma, rickettsia, chlamydia, etc. are mentioned. Especially, it uses suitably for the infection defense of bacteria, fungi, a parasite, or a protozoan.
Examples of diseases in which the compound of the present invention and pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing the same are effective include infections caused by the microorganisms, and immunostimulators or infection prevention or treatment against infections. Excellent effect as an agent. It is also useful for the prevention or treatment of malignant tumors, blood diseases, liver diseases, collagen diseases, renal metabolic diseases, preoperative and / or postoperative infections. Furthermore, it is suitably used for enhancing the immunogenicity of an antigen in a vaccine used for the prevention of infectious diseases caused by the microorganisms, thereby effectively preventing infectious diseases.

The pharmaceutical composition of the present invention can be administered to mammals orally or parenterally (eg, topical administration, intravenous administration, inhalation, etc.). Preferably, it is administered parenterally. Of these, intravenous administration is preferred.
Mammals are humans and non-human mammals (eg, rodents (mouse, rat, hamster, guinea pig, etc.), rabbits, cats, dogs, pigs, monkeys, sheep, cows, horses, etc.), preferably Human.

  The dosage of the pharmaceutical composition of the present invention varies depending on the administration subject, disease, symptom, dosage form, administration route, etc. For example, the dosage when administered to an adult patient (body weight: about 60 kg) is the active ingredient. The amount of (the compound (1) of the present invention) is usually preferably in the range of about 0.0001 mg to 2000 mg, more preferably about 0.001 to 1000 mg per day. This amount can be administered in one to several divided doses.

  Compound (1) of the present invention (preferably Compound (I), Compound (II), Compound (III), or Compound (IV)) and a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, etc. As preferred administration subjects, patients with diseases associated with the above-mentioned Toll-like receptor 4 are preferred. The compound of the present invention (preferably Compound (I), Compound (II), Compound (III), or Compound (IV)) can be used, for example, in patients with reduced immunity, allergic patients, and infection of the above microorganisms. Or is suitably administered for treatment to a patient who is developing an infectious disease caused by a microorganism. Moreover, when infection by microorganisms is not materialized, it can administer as a preventive agent.

  Compound (1) of the present invention (preferably Compound (I), Compound (II), Compound (III) or Compound (IV)) and a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, When used for enhancing the immunogenicity of an antigen in a vaccine or the like, the type of the antigen is not particularly limited. In addition, the amount of the compound of the present invention or a pharmaceutically acceptable salt thereof may be appropriately set according to the type of antigen.

  Toll-like receptor 4 activator, immunostimulant, and antiallergic agent comprising the compound (I), compound (II), compound (III) or compound (IV), or a pharmaceutically acceptable salt thereof as an active ingredient In addition, a preventive or therapeutic agent for diseases associated with Toll-like receptor 4 and a preventive or therapeutic agent for infection prevention are also encompassed in the present invention. An immunogenicity enhancer for an antigen comprising the compound (I), compound (II), compound (III) or compound (IV), or a pharmaceutically acceptable salt thereof as an active ingredient is also encompassed in the present invention. The agent of the present invention may consist only of the compound (I), compound (II), compound (III) or compound (IV), which is an active ingredient, or a pharmaceutically acceptable salt thereof. A pharmaceutically acceptable carrier. The agent of this invention, its preferable aspect, etc. are the same as the pharmaceutical composition mentioned above.

  Compound (1) of the present invention (preferably Compound (I), Compound (II), Compound (III) or Compound (IV)) or a pharmaceutically acceptable salt thereof is generally used in the pharmaceutical field. The method can be used (hereinafter also referred to as a combination) in combination with one or more other drugs (hereinafter also referred to as a combined drug).

  The administration timing of the compound (1) of the present invention (preferably compound (I), compound (II), compound (III) or compound (IV)) or a pharmaceutically acceptable salt thereof, and a concomitant drug is not limited, These may be administered to the administration subject as a compounding agent, or both preparations may be administered simultaneously or at regular intervals. Moreover, you may use as a pharmaceutical which is a kit which consists of the pharmaceutical composition of this invention, and a concomitant drug. The dose of the concomitant drug may be determined according to the dose used clinically, and can be appropriately selected depending on the administration subject, disease, symptom, dosage form, administration route, administration time, combination and the like. The administration form of the concomitant drug is not particularly limited as long as the compound (1) of the present invention or a pharmaceutically acceptable salt thereof and the concomitant drug are combined.

  The concomitant drug is not particularly limited. For example, when the compound (1) of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound is used for enhancing the immunogenicity of an antigen. A vaccine containing the above antigen is preferred.

  Next, an example of a method for producing the compound (1) of the present invention will be described. However, the present invention is not limited to these production methods, and it goes without saying that the compounds of the present invention can be produced by appropriately changing the following production methods. In the production of the compound of the present invention, the order of the reaction can be changed as appropriate, and the reaction may be carried out from a process or substitution site that seems to be rational.

  In addition, for example, a substituent conversion (substituent conversion or further substituent modification, for example, oxidation or reduction of a substituent) step may be appropriately inserted between the steps. When there is a reactive functional group, protection and deprotection may be appropriately performed. Moreover, in order to accelerate | stimulate progress of reaction, reagents other than the illustrated reagent can be used suitably, and reaction (for example, reaction in nitrogen atmosphere) can be performed on anhydrous conditions as needed.

For example, compounds such as compound (I), compound (II), compound (III) and compound (IV) can be produced from, for example, funiculosin. For example, a method of hydrogen reduction of funiculosin, a method of derivatizing funiculosin by various methods, a method of hydrogenating and reducing funiculosin to obtain a skeleton, and then derivatizing by various methods, After subsequent reduction with hydrogen to obtain a skeleton portion, it can be produced by a method of further derivatization.
Funiculosin has a structure represented by the following formula. In the general formula (1), n is 0, R ¹ is a methyl group, R ² is a hydrogen atom, and R ³ is a group represented by the above formula (3-1). A compound in which R ⁴ , R ⁵ , R ⁶ , and R ⁷ are hydroxyl groups.

  Funiculosin can be synthesized by a known method (for example, the method described in Ando et al. (1969) J. Antibiot. (Tokyo). 22 (5): 189-94). Funiculosin used in the examples was obtained from Novartis.

  Examples of derivatization include acetylation, esterification, alkylation, acetalization and the like. The method and conditions for derivatization can be performed by known methods and conditions.

  For example, the compound (I) of the present invention can be obtained, for example, by the production method shown below.

  In the above reaction, funiculosin is usually reduced in a solvent or without a solvent in a hydrogen atmosphere and in the presence of a hydrogenation catalyst.

  Any solvent may be used as long as it is inert to the reaction. For example, tetrahydrofuran (THF), 1,4-dioxane, dimethyl ether (DME), diethyl ether, benzene, toluene, xylene, dimethylformamide ( DMF), dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP), methanol, ethanol, 1-propanol, 2-propanol, acetic acid, water, etc. may be used alone or in admixture of two or more. it can. Of these, THF, acetic acid, water and the like are preferable.

  Although the usage-amount of a solvent is not limited, For example, it is preferable to set it as about 10-100 mass times with respect to the mass of funiculosin.

  The hydrogenation catalyst is not particularly limited, and any metal having hydrogenation ability such as iron, cobalt, nickel, ruthenium, palladium, iridium, platinum, copper, etc. may be used, and it can be used regardless of its form. Among these, palladium is preferable, and palladium hydroxide is more preferable. The amount of the hydrogenation catalyst used is preferably about 0.01 to 50 times by mass, more preferably about 0.1 to 5 times by mass with respect to the mass of funiculosin.

  The reaction is preferably carried out at a temperature between room temperature and the boiling point of the solvent used, for example, preferably at about 30 to 100 ° C. The reaction time is usually about 1 to 150 hours. In the present specification, room temperature means about 1 to 40 ° C.

  Compound (II) of the present invention can be obtained, for example, by the production method shown below.

Process 1
In the reaction described above, funiculosin (represented by the above formula 1) is usually reacted in the presence of an acetalizing reagent and an acid catalyst in a solvent or without a solvent. By this reaction, a compound represented by the above formula 2 (hereinafter also referred to as compound 2) is obtained.

  As the solvent used in the above reaction, a ketone or aldehyde that forms the target acetal, for example, acetone or the like can be used. Any of those inert to the reaction may be used. For example, tetrahydrofuran (THF), 1,4-dioxane, dimethyl ether (DME), diethyl ether, benzene, toluene, xylene, dimethylformamide (DMF), dimethylacetamide ( DMA), N-methyl-2-pyrrolidone (NMP), dichloromethane and chloroform can be used alone or in admixture of two or more. Among these, THF, dichloromethane, chloroform, or a mixed solvent of two or more of these is preferable.

  Although the usage-amount of a solvent is not limited, For example, it is preferable to set it as about 10-100 mass times with respect to the mass of funiculosin.

  The acetalizing reagent is not particularly limited, and may be any aldehyde, ketone, or hemiacetal type or acetal type derivative of aldehyde or ketone. Among these, an aldehyde or an acetal type reactant is preferable. In the production of compound (II), it is preferable to use 2,2, -dimethoxypropane or the like as the acetalizing reagent. The amount of the acetalizing reagent used is preferably about 1 to 100 times, more preferably about 1 to 10 times the mass of funiculosin.

  The acid catalyst is not particularly limited, but Bronsted acids such as hydrogen chloride (or hydrochloric acid), hydrogen bromide, p-toluenesulfonic acid, 10-camphorsulfonic acid (CSA); ammonium bromide, pyridinium hexafluoroantimonate, etc. Ammonium salts; Lewis acids such as trimethylsilyl trifluoromethanesulfonate (TfOH), boron trifluoride etherate, titanium tetrachloride, scandium triflate; solid catalysts such as montmorillonite and HZSM-5 zeolite can also be used. Preferably, 10-camphorsulfonic acid (CSA) or the like is used.

  The reaction is preferably performed at a temperature between −20 ° C. and the boiling point of the solvent used, for example, preferably at about 0-40 ° C. The reaction time is usually about 10 minutes to 10 hours.

Process 2
In the above reaction, compound 2 is usually reacted in a solvent or without solvent in the presence of a halogenated or sulfonate ester having a substituent and a base catalyst. By this reaction, a compound represented by the above formula 3 (hereinafter also referred to as compound 3) is obtained.

  Any solvent may be used as long as it is inert to the reaction. For example, tetrahydrofuran (THF), 1,4-dioxane, dimethyl ether (DME), diethyl ether, benzene, toluene, xylene, dimethylformamide ( DMF), dimethylacetamide (DMA), and N-methyl-2-pyrrolidone (NMP) can be used alone or in admixture of two or more. Of these, THF, DMF and the like are preferable.

  Although the usage-amount of a solvent is not limited, For example, it is preferable to set it as about 5-100 mass times with respect to the mass of the compound 2. FIG.

Although the halide or sulfonate having a substituent is not particularly limited, examples of the leaving group include chloride ion, bromide ion, iodide ion methanesulfonate, p-toluenesulfonate, trifluoromethanesulfonate, and the like. It is possible to use a compound containing
Examples of the substituent contained in the halide or sulfonic acid ester include a hydroxyl group, a carbonyl group, a carboxyl group, and an amino group, with a hydroxyl group being preferred. The amount of the halide or sulfonic acid ester to be used is preferably about 0.5 to 100 times by mass, more preferably about 1 to 10 times by mass with respect to the mass of Compound 2.
The halogenated or sulfonic acid ester having a substituent used in Step 2 is preferably 1-bromoethanol or the like.

The base catalyst is not particularly limited, but may be an alkali metal base such as Na, NaH, KH, NaOH, KOH, Na ₂ CO ₃ , K ₂ CO _3, or 1,8-diazabicyclo [5,4,0] undec-7. - it is possible to use ene _{(DBU), n-Bu2SnO,} Ni (acac) 2, Ag 2 O, AgOTf, base or catalyst such as _Et 4 NF. Preferably, it is an alkali metal base, more preferably K ₂ CO ₃ or the like.

  The reaction is preferably performed at a temperature between −20 ° C. and the boiling point of the solvent used, for example, preferably about 0 to 70 ° C. The reaction time is usually about 1 to 100 hours.

Process 3
In the above reaction, compound 3 is usually reacted in the presence of an oxidizing agent in a solvent or without a solvent. It is also possible to carry out the reaction using a reoxidant in combination with the main oxidant. It is also possible to carry out the reaction from alcohol to aldehyde and from aldehyde to carboxylic acid in two stages with different oxidizing agents. According to step 3, the compound (II) is obtained.

  Any solvent can be used in the above reaction as long as it is inert to the reaction. For example, benzene, toluene, xylene, dimethylformamide (DMF), dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP) , Dichloromethane, chloroform, tetrahydrofuran (THF), 1,4-dioxane, dimethyl ether (DME), diethyl ether and water can be used alone or in admixture of two or more. Of these, dichloromethane, chloroform or dichloromethane, a mixed solvent of chloroform and water, and the like are preferable.

  Although the usage-amount of a solvent is not limited, For example, it is preferable to set it as about 5-100 mass times with respect to the mass of the compound 3.

The oxidizing agent is not particularly limited, but an oxidizing agent such as 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) or TEMPO and sodium hypochlorite, trichloroisocyanuric acid ( It is also possible to react with a catalytic amount of TEMP by combining TCC) or PhI (OAc) ₂ . As other oxidizing agents, peroxides such as peracetic acid, reactions with hexavalent chromates under acidic conditions, permanganates, ruthenium tetroxide and perruthenates are also used. Among them, a combination of TEMPO and PhI (OAc) ₂ is preferably used. The amount of the oxidizing agent used is preferably about 0.1 to 100 times by mass, more preferably about 1 to 10 times by mass with respect to the mass of compound 3.

  The reaction is preferably performed at a temperature between −20 ° C. and the boiling point of the solvent used or the decomposition temperature of the oxidizing agent, for example, preferably about 0 to 70 ° C. The reaction time is usually about 0.1 to 100 hours.

Compounds (III) and (IV) of the present invention can be obtained, for example, by the production method shown below.
The reaction from compound 3 to compound 8 is carried out by reduction in the presence of a hydrogenation catalyst in a hydrogen atmosphere in a solvent or without a solvent in the same manner as in the reaction for producing compound (1) from funiculosin.

Any solvent may be used as long as it is inert to the reaction. For example, tetrahydrofuran (THF), 1,4-dioxane, dimethyl ether (DME), diethyl ether, benzene, toluene, xylene, dimethylformamide ( DMF), dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP), methanol, ethanol, 1-propanol, 2-propanol, acetic acid, water, etc. may be used alone or in admixture of two or more. it can. Of these, THF, acetic acid, water and the like are preferable.

  Although the usage-amount of a solvent is not limited, For example, it is preferable to set it as about 10-100 mass times with respect to the mass of funiculosin.

  The hydrogenation catalyst is not particularly limited, and any metal having hydrogenation ability such as iron, cobalt, nickel, ruthenium, palladium, iridium, platinum, copper, etc. may be used, and it can be used regardless of its form. Among these, palladium is preferable, and palladium hydroxide is more preferable. The amount of the hydrogenation catalyst used is preferably about 0.01 to 50 times by mass, more preferably about 0.1 to 5 times by mass with respect to the mass of funiculosin.

  The reaction is preferably carried out at a temperature between room temperature and the boiling point of the solvent used, for example, preferably at about 30 to 100 ° C. The reaction time is usually about 1 to 150 hours. In the present specification, room temperature means about 1 to 40 ° C.

  The reactions from Compound 3 to Compound 6 and from Compound 8 to Compound 11 are all phosphorylation reaction steps, and the same method can be used.

  Although it does not specifically limit as phosphorylation, The phosphoric acid triester method, the phosphorous acid triester method, the phosphoramidite method, the diamidite method, the H-phosphonate method, etc. can be used.

  Any solvent can be used in the above reaction as long as it is inert to the reaction. For example, tetrahydrofuran (THF), 1,4-dioxane, dimethyl ether (DME), diethyl ether, benzene, toluene, xylene, chloroform, dichloromethane , Dichloroethane, dimethylformamide (DMF), dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP), acetonitrile, pyridine and the like can be used alone or in admixture of two or more.

  Although the usage-amount of a solvent is not limited, For example, it is preferable to set it as about 2-200 mass times with respect to the mass of starting materials, such as the compound 3 or the compound 8.

  The reaction is preferably performed at a temperature between −50 ° C. and the boiling point of the solvent used, for example, preferably at about −20 to 100 ° C. The reaction time is usually about 0.5 to 150 hours. In the present specification, room temperature means about 1 to 40 ° C.

  The reaction of compound 6 to compound (III) and compound 11 to compound (IV) is the same deprotection step, and a method of simultaneously cleaving an acetal group and a phosphoric acid protecting group under acidic conditions, A method of performing each deprotection step by step can be used.

  When the cleavage of the acetal group and the cleavage of the protecting group of phosphoric acid are performed simultaneously under acidic conditions, the acidic reagent is not particularly limited, but various Lewis acids including silyl-based Lewis acids can be used. In the case of cleaving stepwise, a method such as cleaving the acetal group under various acidic conditions including protonic acid and Lewis acid and cleaving the protecting group of phosphoric acid under reducing conditions or acidic conditions can be used. .

  Any solvent can be used as long as it is inert to the reaction. For example, chloroform, dichloromethane, dichloroethane, benzene, toluene, xylene, tetrahydrofuran (THF), 1,4-dioxane, dimethyl ether (DME), Diethyl ether, dimethylformamide (DMF), dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP), acetonitrile and the like can be used alone or in admixture of two or more.

  Although the usage-amount of a solvent is not limited, For example, it is preferable to set it as about 2-200 mass times with respect to the mass of starting materials, such as the compound 6 or the compound 11.

  The reaction is preferably performed at a temperature between −50 ° C. and the boiling point of the solvent used, for example, preferably at about −20 to 100 ° C. The reaction time is usually about 0.5 to 150 hours. In the present specification, room temperature means about 1 to 40 ° C.

For the compound (I), the compound (II), the compound (III) or the compound (IV), and the compound represented by the general formula (1) other than the compounds 2 and 3, the above methods and raw materials are known methods. It can manufacture by changing suitably with a raw material.
For example, the compound (I), the compound (II), the compound (III), the compound (IV), or an intermediate thereof may be further derivatized by a method known per se to obtain another compound represented by the above formula (1). Compounds can be produced.

  The compound of the present invention can be obtained by the above production method. Isolation and purification of intermediates and products in the above production method can be performed by appropriately combining methods used in ordinary organic synthesis, for example, filtration, extraction, washing, drying, concentration, crystallization, various chromatography, and the like. it can. The intermediate can also be subjected to the next reaction without particular purification.

  The raw material compound or intermediate in the above production method may exist in the form of a salt such as hydrochloride depending on the reaction conditions and the like, but can be used as it is or in a free form. When the raw material compound or intermediate is obtained in the form of a salt, and the raw material compound or intermediate is to be used or obtained in a free form, these are dissolved or suspended in a suitable solvent, for example, a base (for example, sodium bicarbonate It can be converted into a free form by neutralization with an aqueous solution or the like.

  When it is desired to obtain a salt of the compound (1) of the present invention (for example, the compound (I), the compound (II), the compound (III) or the compound (IV)), the salt of the compound (1) is obtained as it is. What is necessary is just to refine | purify and when a compound (1)) is obtained in a free form, what is necessary is just to melt | dissolve or suspend this compound in a suitable solvent, and to form a salt by adding an acid or a base. In addition, compound (1) or a pharmaceutically acceptable salt thereof may exist in the form of a solvate with water or various solvents, and these solvates are also encompassed in the present invention.

EXAMPLES Next, although an Example is given and this invention is demonstrated further more concretely, this invention is not limited at all by these Examples.
Unless otherwise specified, “%” means mass%, and “solvent ratio” means volume ratio.

The physicochemical data of each compound in the examples was measured by the following instruments.
¹ HNMR: JEOL ECA 500 (500 MHz)
MS: BRUKER micrOTOF-Q II-HC (registered trademark) (ESI-TOF-MS method)
Applied Biosystem Voyager (registered trademark) (MALDI-TOF-MS)
In addition, although what is used normally is used as a description of the multiplicity of a signal, br expresses that it is a wide signal apparently.

Purification of each compound was performed using a reverse phase (ODS) column (trade name Cosmosil 5C18-AR-300) manufactured by Nacalai Tesque, under the following apparatus and conditions.
Equipment: Shimadzu LC10A (Shimadzu Corporation)
Detection wavelength: 220 nm
Solvent: Solution A: 0.1% TFA in water, Solution B: CH ₃ CN (B solution concentration 70%)
Column temperature: room temperature (rt)
Flow rate: 1 mL / min

Each compound was named using ChemBioDraw Ver. 11.0 (Cambridge Soft).
In Examples, THF is tetrahydrofuran, (+)-CSA is (+)-10-camphorsulfonic acid, DMF is dimethylformamide, LHMDS is lithium hexamethyldisilazide, and TEMPO is 2,2,6,6. -Tetramethylpiperidine 1-oxyl, respectively.

<Example 1>
Production of funiculosin derivative (compound (I))

Funiculosin (Compound 1) 1 mg and palladium hydroxide / carbon (Pd (OH) _{2 /} C) (2.0 mg, Pd 20%, about 50% water wet) with THF (1 mL) and water (0.1 mL) The mixture was suspended in a mixed solvent with acetic acid (0.05 mL), and stirred for 5 days under a hydrogen atmosphere (30 kg / cm ² ). Pd (OH) ₂ / C was removed by a membrane filter, and the filtrate was concentrated under reduced pressure. The operation of adding azeotropically with adding toluene was repeated three times, and purified by HPLC, whereby a funiculosin derivative having the structure represented by the above formula (I) (chemical name: 4-hydroxy-1-methyl-5-((( 1R, 2R, 3R, 4S, 5S) -2,3,4,5-tetrahydroxycyclopentyl) -3-((8S, 10R) -4,6,8,10-tetramethyldodecyl) pyridine-2 (1H ) -One (4-hydroxy-1-methyl-5-((1R, 2R, 3R, 4S, 5S) -2,3,4,5-tetrahydroxycyclopentyl) -3-((8S, 10R) -4,6 , 8,10-tetramethyldodecyl) pyridin-2 (1H) -one)) (compound (I)) was obtained as a yellow oil.
The NMR value and MS analysis value of compound (I) are shown below.
¹ H-NMR (CDCl ₃ ) δ: 7.36 (s, 1H), 4.37 (s, 1H), 4.12 (q, 1H, J = 7.1 Hz), 3.72 (q, 1H, J = 7.1 Hz), 3.50 ( d, 2H, J = 6.3 Hz), 2.74 (t, 1H, J = 4.1 Hz), 2.41 (s, 1H), 2.07 (d, 1H, J = 25.6 Hz), 1.50-1.44 (m, 3H), 1.29-1.25 (m, 2H), 1.17-0.91 (m, 4H), 0.88-0.77 (m, 8H).
HRMS (ESI-QTOF MS, positive) Calcd. For C ₂₇ H ₄₈ NO ₆ [M + Na] ⁺ : 504.3301, Found 504.3303

<Example 2>
Production of Funiculosin Derivative (Compound (II)) A funiculosin derivative having the structure represented by the above formula (II) was produced by the following method (Steps 1 to 3).

(Process 1)
3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2-yl) -5-methyl-3,6-dihydro-2H-pyran- 2-yl) -4-hydroxy-1-methyl-5-((3aR, 3bS, 6aS, 7r, 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4-d '] bis ([1,3] dioxol) -7-yl) pyridin-2 (1H) -one (3-((2R, 6S) -6-((4S, 6R, E)- 4,6-dimethyloct-2-en-2-yl) -5-methyl-3,6-dihydro-2H-pyran-2-yl) -4-hydroxy-1-methyl-5-((3aR, 3bS, 6aS, 7r, 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4-d '] bis ([1,3] dioxole) -7-yl) pyridin- 2 (1H) -one) (Compound 2)

  Funiculosin (Compound 1) (20.0 mg, 41.5 μmol) and (+)-CSA (2.90 mg, 12.5 μmol) were dissolved in a mixed solvent of 2,2-dimethoxypropane (415 μL) and anhydrous dichloromethane (415 μL). Stir at room temperature for 80 minutes. Saturated sodium hydrogen carbonate was added to the reaction solution, stirred, and extracted with chloroform. The organic layer was washed successively with a saturated aqueous sodium bicarbonate solution and a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The desiccant was removed and the filtrate was concentrated under reduced pressure. The resulting residue was purified by medium pressure silica gel column chromatography (chloroform / methanol = 20: 1), and the compound 2 represented by the above formula was converted to a yellow solid (11.7 mg, 50%).

The analytical value of Compound 2 is shown below.
HRMS (ESI-QTOF MS, positive) Calcd. For C ₃₃ H ₄₉ NO ₇ Na [M + Na] ⁺ : 594.3401, Found 594.3404
¹ H-NMR (500 MHz, CDCl ₃ ) δ: 9.84 (s, 1H, C ₁₉ -O H ), 7.68 (s, 1H, C ₂₀ - H ), 5.74 (d, 1H, J = 6.3 Hz, C ₁₄ - H ), 5.33 (d, 1H, J = 9.5 Hz, C ₁₄ - H ), 5.12 (dd, 1H, J = 10.5, 3.1 Hz, C ₁₆ - H ), 4.70 (t, 1H, J = 5.1 Hz, C _{24 or 27} - H ), 4.67 (t, 1H, J = 4.9 Hz, C _{24 or 27} - H ), 4.56 (s, 1H, C ₁₁ - H ), 4.50-4.46 (m, 2H, C ₂₅ - H , C ₂₆ - H ), 3.48 (s, 3H, NC H ₃ ), 3.41 (t, 1H, J = 5.5 Hz, C ₂₃ - H ), 2.51-2.46 (m, 2H, C ₁₅ - H ₂ ), 2.28-2.22 (m, 1H, C ₆ - H ), 1.63 (d, 6H, J = 3.6 Hz, C ₂₉ - H ₃ , C ₃₀ - H ₃ ), 1.57 (d, 3H, J = 1.3 Hz, C ₁₀ - H ₃ ), 1.50 (d, 3H, J = 1.1 Hz, C ₁₃ - H ₃ ), 1.33 (d, 8H, J = 8.9 Hz, C ₃₂ - H ₃ , C ₃₃ - H _{3 ,} C ₅ - H ₂ ), 1.25-1.22 (m, 1H, C ₃ - H ), 1.09-1.05 (m, 2H, C ₂ - H ₂ ), 0.94 (d, 3H, J = 6.7 Hz, C ₁ - H ₇ ), 0.84 (m, 6H, J = 9.5, 4.7 Hz, C ₁ - H ₃ , C ₄ - H ₃ ).

(Process 2)
3-((2R, 6S) -6-((4S, 6R, E) -4,6-Dimethyloct-2-en-2-yl) -5-methyl-3,6-dihydro-2H-pyran- 2-yl) -4- (2-hydroxyethoxy) -1-methyl-5-((3aR, 3bS, 6aS, 7r, 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1 , 2-d: 3,4-d '] bis ([1,3] dioxol) -7-yl) pyridin-2 (1H) -one (3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2-yl) -5-methyl-3,6-dihydro-2H-pyran-2-yl) -4- (2-hydroxyethoxy) -1-methyl -5-((3aR, 3bS, 6aS, 7r, 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4-d '] bis ([1,3] dioxole) -7-yl) pyridin-2 (1H) -one) (compound 3)

  Compound 2 (10.0 mg, 17.5 μmol) obtained in Step 1 and potassium carbonate (29.0 mg, 210 μmol) are suspended in a mixed solvent of anhydrous DMF (100 μL) and anhydrous THF (100 μL), and the mixture is 50 at 40 ° C. Stir for minutes. 2-Bromoethanol (12.5 μL, 175 μmol) was added to the reaction solution and stirred for 2 days. 1M hydrochloric acid was added to the reaction solution, stirred, and extracted with chloroform. The organic layer was washed successively with 1M hydrochloric acid and saturated aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The desiccant was removed and the filtrate was concentrated under reduced pressure. The resulting residue was purified by medium pressure silica gel column chromatography (hexane / ethyl acetate = 2: 1) to give compound 3 represented by the above formula as a yellow solid (5.9 mg). , 54%).

The analytical value of Compound 2 is shown below.
HRMS (ESI-QTOF MS, positive) Calcd. For C ₃₅ H ₅₃ NO ₈ Na [M + Na] ⁺ : 638.3663, Found 638.3677
¹ H-NMR (500 MHz, CDCl ₃ ) δ: 7.75 (s, 1H, C ₂₁ - H ), 5.73 (d, 1H, J = 6.3 Hz, C ₁₄ - H ), 5.23 (d, 1H, J = 9.5 Hz, C ₈ - H ), 5.09 (dd, 1H, J = 10.5, 3.1 Hz, C ₁₆ - H ), 4.66 (t, 1H, J = 5.1 Hz, C ₂₄ - H ), 4.52 (t, 1H , J = 4.9 Hz, C ₂₇ - H ), 4.53-4.50 (m, 3H, C ₂₅ - H , C ₂₆ - H , C ₁₁ - H ), 4.23 (m, 1H, C ₃₄ - H ), 4.00 ( _{m, 1H, C 34 - H} ), 3.94-3.87 (m, 2H, C 35 - H 2), 3.51 (s, 3H, NC H 3), 3.50 (t, 1H, J = 5.5 Hz, C 23 - H), 3.15 (b, H , C 15 - H), 2.48-2.42 (m, 1H, C 6 - H), 2.36 (b, H, C 35 -O H) 1.88 (b, H, C 15 - H ) 1.63 (d, 6H, J = 3.6 Hz, C ₂₉ - H ₃ , C ₃₀ - H ₃ ), 1.57 (d, 3H, J = 1.3 Hz, C ₁₀ - H ₃ ), 1.50 (d, 3H, J = 1.1 Hz, C ₁₃ - H ₃ ), 1.33 (d, 8H, J = 8.9 Hz, C ₃₂ - H ₃ , C ₃₃ - H _{3 ,} C ₅ - H ₂ ), 1.25-1.22 (m, 1H, C ₃ - H ), 1.09-1.05 (m, 2H, C ₂ - H ₂ ), 0.94 (d, 3H, J = 6.7 Hz, C ₇ - H ₃ ), 0.84 (m, 6H, J = 9.5, 4.7 Hz, C ₁ - H ₃ , C ₄ - H ₃ ).
¹³ C-NMR (500 MHz, CDCl ₃ ) δ: 165.32 (C ₁₉ ), 162.73 (C ₁₇ ), 142.64 (C ₂₁ ), 137.91 (C ₈ ), 133.65 (C ₁₂ ), 131.97 (C ₁₈ ), 122.56 (C ₁₄ ), 120.06 (C ₂₀ ), 112.52 (C ₉ ), 108.12 (C ₂₈ , C ₃₁ ), 86.69 (C ₁₁ ), 83.05 (C ₂₄ , C ₂₇ ), 78.96 (C ₂₅ , C ₂₆ ), 76.77 (C ₃₄ ), 69.75 (C ₁₆ ), 62.22 (C ₃₅ ), 44.86 (C ₃ ), 38.31 (C ₂₂ ), 36.00 (C ₂₃ ), 32.00 (C ₅ ), 29.81 (C ₆ ), 29.00 ( C ₂ ), 28.15 (C ₁₅ ), 25.63 (C ₂₉ , C ₃₀ ), 24.26 (C ₃₂ , C ₃₃ ), 20.66 (C ₇ ), 19.60 (C ₄ ), 19.22 (C ₁₀ ), 11.23 (C ₁ ).

(Process 3)
2-((3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2-yl) -5-methyl-3,6-dihydro- 2H-pyran-2-yl) -1-methyl-2-oxo-5-((3aR, 3bS, 6aS, 7r, 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1, 2-d: 3,4-d '] bis ([1,3] dioxole) -7-yl) -1,2-dihydropyridin-4-yl) oxy) acetic acid (2-((3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2-yl) -5-methyl-3,6-dihydro-2H-pyran-2-yl) -1- methyl-2-oxo-5-((3aR, 3bS, 6aS, 7r, 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4-d '] bis ( [1,3] dioxole) -7-yl) -1,2-dihydropyridin-4-yl) oxy) acetic acid) (compound (II))

Compound 3 (2.0 mg, 3.30 μmol), TEMPO (1.02 mg, 6.62 μmol) and iodobenzene diacetate (4.19 mg, 13.2 μmol) obtained in step 2 were dissolved in dichloromethane (400 μL) and stirred at room temperature for 3 hours. did. Water (100 μL) was added to the reaction solution and stirred for 1 hour. A 10% aqueous sodium thiosulfate solution was added to the reaction solution, and purification was performed by gel permeation chromatography (methanol) to obtain compound (II) represented by the above formula as a yellow oil.
The analytical value of compound (II) is shown below.
HRMS (ESI-QTOF MS, positive) Calcd. For C ₃₅ H ₅₁ NO ₉ Na [M + Na] ⁺ : 652.3456, Found 652.3477

<Example 3>
A method for synthesizing the other compound represented by the general formula (1) is shown below.

4-Acetoxy-1-methyl-5-((1R, 2R, 3R, 4S, 5S) -2,3,4,5-tetraacetoxycyclopentyl) -3-((8S, 10R) -4,6,8 , 10-Tetramethyldodecyl) pyridin-2 (1H) -one (4-acetoxy-1-methyl-5-((1R, 2R, 3R, 4S, 5S) -2,3,4,5-tetraacetoxycyclopentyl)- 3-((8S, 10R) -4,6,8,10-tetramethyldodecyl) pyridin-2 (1H) -one) (compound 4)

  Compound (I) (1 mg) synthesized in Example 1 was dissolved in a mixed solvent of acetic anhydride (60 μL) and anhydrous pyridine (60 μL) and stirred at room temperature for 2 days. 1 M hydrochloric acid was added to the reaction solution, stirred, and extracted with chloroform. The organic layer was washed successively with 1 M hydrochloric acid and saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The desiccant was filtered off and concentrated under reduced pressure, and the resulting residue was purified by HPLC to obtain Compound 4 represented by the above formula.

The analytical value of Compound 4 is shown below.
MALDI-TOF MS (positive): m / z = 692.86 [M + H] ⁺ , 714.86 [M + Na] ⁺ ;
HRMS (ESI-QTOF MS, positive) Calcd. For C ₃₇ H ₅₇ NO ₁₁ [M + Na] ⁺ : 714.3829, Found 714.3827
¹ H-NMR (CDCl ₃ ) δ: 7.39 (s, 1H), 5.38 (t, 4H, J = 5.3 Hz), 3.57 (s, 3H), 3.33 (t, 1H, J = 6.3 Hz), 2.37- 2.36 (m, 2H), 2.33 (s, 3H), 2.07 (s, 6H), 2.03 (s, 6H), 1.51-0.89 (m, 16H), 0.88-0.78 (m, 15H).
¹³ C-NMR (CDCl ₃ ) δ: 169.22, 167.95, 162.71, 155.27, 138.23, 138.23, 124.03, 105.92, 71.37, 69.92, 47.13, 46.22, 45.69, 45.43, 44.80, 44.53, 38.62, 38.13, 37.60, 31.88, 31.76, 30.70, 30.28, 30.06, 29.88, 27.47, 26.21, 25.27, 20.64, 20.46, 20.09, 20.00, 19.35, 19.27, 19.09, 11.59, 11.46.

<Example 4>
(1R, 2R, 3S, 4S, 5R) -5- (4-acetoxy-5-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-ene- 2-yl) -5-methyl-3,6-dihydro-2H-pyran-2-yl) -1-methyl-6-oxo-1,6-dihydropyridin-3-yl) cyclopentane-1,2,3 , 4-Tetrayl tetraacetate ((1R, 2R, 3S, 4S, 5R) -5- (4-acetoxy-5-((2R, 6S) -6-((4S, 6R, E) -4,6- dimethyloct-2-en-2-yl) -5-methyl-3,6-dihydro-2H-pyran-2-yl) -1-methyl-6-oxo-1,6-dihydropyridin-3-yl) cyclopentane- 1,2,3,4-tetrayl tetraacetate) (Compound 5)

Funiculosin (Compound 1) (5.0 mg, 10.2 μmol) was dissolved in a mixed solvent of acetic anhydride (60 μL) and anhydrous pyridine (60 μL) and stirred at room temperature for 1.5 hours. 1 M hydrochloric acid was added to the reaction solution and stirred, followed by extraction with ethyl acetate. The organic layer was washed successively with 1 M hydrochloric acid and saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The desiccant was removed by filtration and the filtrate was concentrated under reduced pressure. The resulting residue was purified by medium pressure silica gel column chromatography (chloroform) to obtain Compound 5 represented by the above formula as a white solid.
The analytical value of Compound 5 is shown below.
ESI-Mass (positive) m / z = 702.94 [M + H] ⁺ , 724.94 [M + Na] ⁺ ;
HRMS (ESI-QTOF MS, positive) Calcd. For C ₃₇ H ₅₁ NO ₁₂ [M + Na] ⁺ : 724.3309, Found 724.3309

<Example 5>
Dibenzyl (2-((3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2-yl) -5-methyl-3,6- Dihydro-2H-pyran-2-yl) -1-methyl-2-oxo-5-((3aR, 3bS, 6aS, 7r, 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [ 1,2-d: 3,4-d '] bis ([1,3] dioxol) -7-yl) -1,2-dihydropyridin-4-yl) oxy) ethyl) phosphate (dibenzyl (2-(( 3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2-yl) -5-methyl-3,6-dihydro-2H-pyran-2 -yl) -1-methyl-2-oxo-5-((3aR, 3bS, 6aS, 7r, 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4 -d '] bis ([1,3] dioxole) -7-yl) -1,2-dihydropyridin-4-yl) oxy) ethyl) phosphate) (compound 6)

Compound 3 (1.0 mg, 1.62 μmol) prepared in Step 2 in the preparation of the funiculosin derivative (Compound (II)) of Example 2 was dissolved in anhydrous THF (200 μL) and stirred at 0 ° C. for 10 minutes, and LHMDS (1.0 M in THF, 9.7 μL, 9.7 μmol) was added, and the mixture was stirred at 0 ° C. for 1 hour. Tetrabenzyl pyrophosphate (5.25 mg, 9.7 μmol) was added to the reaction solution, and the mixture was stirred at 0 ° C. for 1 hour, and further stirred at room temperature for 24 hours. To the reaction solution was added 1 M hydrochloric acid, and the mixture was stirred and extracted with dichloromethane. The organic layer was washed successively with 1 M hydrochloric acid and saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The desiccant was removed and the filtrate was concentrated under reduced pressure. The resulting residue was purified by medium pressure silica gel column chromatography (hexane / ethyl acetate / triethylamine = 70: 10: 0.1) to give compound 6 represented by the above formula yellow. Obtained as a solid.
The MS analysis value of Compound 6 is shown below.
ESI-TOF-MS (positive): m / z = 898.43 [M + Na] ⁺

<Example 6>
Dibenzyl (2-((3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2-yl) -5-methyl-3,6- Dihydro-2H-pyran-2-yl) -1-methyl-2-oxo-5-((1r, 2R, 3R, 4S, 5S) -2,3,4,5-tetrahydroxycyclopentyl) -1,2 -Dihydropyridin-4-yl) oxy) ethyl) phosphate (dibenzyl (2-((3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2 -yl) -5-methyl-3,6-dihydro-2H-pyran-2-yl) -1-methyl-2-oxo-5-((1r, 2R, 3R, 4S, 5S) -2,3, 4,5-tetrahydroxycyclopentyl) -1,2-dihydropyridin-4-yl) oxy) ethyl) phosphate) (compound 7)

Compound 6 (1.3 mg, 1.48 μmol) prepared in Example 5 was dissolved in a mixed solvent of dichloromethane (200 μL), water (40 μL) and TFA (100 μL) and stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the operation of adding toluene and azeotropic drying was repeated 3 times to obtain Compound 7 represented by the above formula as a yellow oil.
The MS analysis value of Compound 7 is shown below.
ESI-TOF-MS (positive): m / z = 796.38 [M + H] ⁺

<Example 7> 2-((3-((2R, 6S) -6-((4S, 6R, E) -4,6-dimethyloct-2-en-2-yl) -5-methyl-3, 6-dihydro-2H-pyran-2-yl) -1-methyl-2-oxo-5-((1r, 2R, 3R, 4S, 5S) -2,3,4,5-tetrahydroxycyclopentyl) -1,2 -dihydropyridin-4-yl) oxy) ethyl dihydrogen phosphate

Compound 6 (2.5 mg, 2.85 μmol) prepared in Example 5 was dissolved in dichloromethane (1.0 mL). The solution was cooled to 0 ° C., TMSBr (2.6 μL, 14.3 μmol) was added dropwise, and the mixture was stirred at room temperature for 6 hours. The reaction solution was cooled to 0 ° C., water (5 μL) was added dropwise, and the mixture was stirred for 5 minutes. The operation of concentrating the reaction solution under reduced pressure, adding toluene and drying azeotropically was repeated three times. The obtained residue was purified by HPLC to obtain Compound (III) represented by the above formula as a white solid.
The analytical value of compound (III) is shown below.
HRMS (ESI-QTOF MS, negative) Calcd. For C ₂₉ H ₄₆ NO ₁₁ P [MH] ^- : 614.2736, Found 614.2742
¹ H-NMR (700 MHz, CD ₃ OD) δ: 7.74 (s, 1H), 5.38 (m, 1H), 5.17 (m, 1H), 4.44 (m, 1H), 4.08 (m, 2H), 4.01 (m, 1H), 3.94 (m, 2H), 3.87 (m, 1H), 3.82 (m, 1H), 3.79 (m, 1H), 3.42 (m, 1H), 3.37 (s, 3H), 2.24 ( m, 1H), 2.03 (m, 1H), 1.52-0.97 (m, 11H), 0.95-0.85 (m, 9H).

<Example 8>
4- (2-hydroxyethoxy) -1-methyl-3-((8S, 10R) -4,6,8,10-tetramethyldodecyl) -5-((3aR, 3bS, 6aS, 7r, 7aR)- 2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4-d '] bis ([1,3] dioxol) -7-yl) pyridin-2 (1H)- ON (4- (2-hydroxyethoxy) -1-methyl-3-((8S, 10R) -4,6,8,10-tetramethyldodecyl) -5-((3aR, 3bS, 6aS, 7r, 7aR) -2 , 2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4-d '] bis ([1,3] dioxole) -7-yl) pyridin-2 (1H) -one) ( Production of compound 8)

Compound 3 (1.5 mg, 2.44 μmol) produced in Step 2 of production of funiculosin derivative (compound (II)) of Example 2 and palladium hydroxide / carbon (2.0 mg, Pd 20%, about 50% water wet) Was suspended in a mixed solvent of THF (800 μL), water (80 μL) and acetic acid (40 μL), and stirred for 2 days under a hydrogen atmosphere of 30 kg / cm ² . Palladium hydroxide / carbon was filtered off with a membrane filter, and the filtrate was concentrated under reduced pressure. The operation of adding toluene and azeotropically drying was repeated three times, and purified by HPLC to obtain Compound 8 represented by the above formula as a yellow oil.
The analytical value of Compound 8 is shown below.
HRMS (ESI-QTOF MS, positive) Calcd. For C ₃₅ H ₅₉ NO ₇ Na [M + Na] ⁺ : 628.4208, Found 628.4192
¹ H-NMR (500 MHz, CDCl ₃ ) δ: 7.69 (s, 1H, C ₂₁ - H ), 4.67 (m, 2H, C ₃₄ - H ₂ ), 4.53 (m, 2H, C ₃₅ - H ₂ ) , 3.94 (m, 4H, C ₂₄ - H , C ₂₅ - H , C ₂₆ - H , C ₂₇ - H ), 3.54 (s, 3H, NC H ₃ ), 3.46 (t, 1H, J = 5.5 Hz, C ₂₃ - H ), 2.51 (m, 2H, C ₁₆ - H ₂ ), 1.94 (b, 1H, O- H ), 1.64 (s, 6H, C ₂₉ - H ₃ , C ₃₀ - H ₃ ), 1.33 (s, 6H, C ₂₉ - H ₃ , C ₃₀ - H ₃ ), 1.52-0.97 (m, 16H, C ₂ - H ₂ , C ₃ - H _, C ₅ - H ₂ , C ₆ - H _, C ₈ - H ₂ , C ₉ - H _, C ₁₁ -H ₂ , C ₁₂ - H _, C ₁₄ - H ₂ , C ₁₅ - H _2, ), 0.88-0.78 (m, 15H, C ₁ - H ₃ , C ₄ - H ₃ , C ₇ - H ₃ , C ₁₀ - H ₃ , C ₁₃ - H ₃ ,)

<Example 9>
2-((1-Methyl-2-oxo-3-((8S, 10R) -4,6,8,10-tetramethyldodecyl) -5-((3aR, 3bS, 6aS, 7r, 7aR) -2 , 2,5,5-Tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4-d '] bis ([1,3] dioxol) -7-yl) -1,2-dihydropyridine-4 -Yl) oxy) acetic acid (2-((1-methyl-2-oxo-3-((8S, 10R) -4,6,8,10-tetramethyldodecyl) -5-((3aR, 3bS, 6aS, 7r , 7aR) -2,2,5,5-tetramethyltetrahydro-3aH-cyclopenta [1,2-d: 3,4-d '] bis ([1,3] dioxole) -7-yl) -1,2- dihydropyridin-4-yl) oxy) acetic acid) (compound 9)

Compound 8 (1.0 mg, 1.65 μmol) prepared in Example 8, TEMPO (0.520 mg, 3.31 μmol), and iodobenzene diacetate (2.14 mg, 6.61 μmol) were dissolved in dichloromethane (300 μL), and the mixture was stirred at room temperature for 1 hour. Stir. Water (100 μL) was added to the reaction solution, and the mixture was stirred for 3 hours. A 10% aqueous sodium thiosulfate solution was added to the reaction solution, and purification was performed by gel permeation chromatography (methanol) to obtain Compound 9 represented by the above formula as a yellow oil.
The MS analysis value of Compound 9 is shown below.
HRMS (ESI-QTOF MS, positive) Calcd. For C ₃₃ H ₄₉ NO ₇ Na [M + Na] ⁺ : 642.3976, Found 642.3967

<Example 10>
2-((1-Methyl-2-oxo-5-((1r, 2R, 3R, 4S, 5S) -2,3,4,5-tetrahydroxycyclopentyl) -3-((8S, 10R) -4 , 6,8,10-Tetramethyldodecyl) -1,2-dihydropyridin-4-yl) oxy) acetic acid (2-((1-methyl-2-oxo-5-((1r, 2R, 3R, 4S, 5S) -2,3,4,5-tetrahydroxycyclopentyl) -3-((8S, 10R) -4,6,8,10-tetramethyldodecyl) -1,2-dihydropyridin-4-yl) oxy) acetic acid) ( Production of compound 10)

Compound 9 produced in Example 9 was dissolved in a mixed solvent of acetonitrile (400 μL), water (100 μL) and TFA (200 μL) and stirred at 0 ° C. for 3 days. The operation of concentrating the reaction solution under reduced pressure, adding toluene and azeotropically drying the mixture was repeated three times to obtain Compound 10 represented by the above formula as a yellow oil.
The MS analysis value of Compound 10 is shown below.
HRMS (ESI-QTOF MS, positive) Calcd. For C ₂₉ H ₅₀ NO ₈ [M + H] ⁺ : 540.3531, Found 540.3545

<Example 11>
dibenzyl (2-((1-methyl-2-oxo-3-((8S, 10R) -4,6,8,10-tetramethyldodecyl) -5-((3aR, 3bS, 6aS, 7r, 7aR) -2 , 2,5,5-tetramethyltetrahydro-7H-cyclopenta [1,2-d: 3,4-d '] bis ([1,3] dioxole) -7-yl) -1,2-dihydropyridin-4-yl ) Oxy) ethyl) phosphate (Compound 11)
Compound 8 (3.0 mg, 4.95 μmol) prepared in Example 8 and tetrazole (3.45 mg, 49.5 μmol) were dissolved in dichloromethane (1.5 mL). The solution was cooled to 0 ° C., dibenzyl N, N-diisopropyl phosphoramidite (4.9 μL, 15.0 μmol) was added, and the mixture was stirred at room temperature for 4 hours. 30% hydrogen peroxide solution (15.2 μL, 149 μmol) was added to the reaction solution, and the mixture was stirred for 3 hours. A 10% aqueous sodium thiosulfate solution was added to the reaction solution, and the mixture was stirred and extracted with chloroform. The organic layer was washed successively with 10% aqueous sodium thiosulfate, saturated aqueous sodium bicarbonate and saturated aqueous sodium chloride and dried over anhydrous sodium sulfate. The desiccant was removed by filtration and dried under reduced pressure. The resulting residue was purified by silica gel column chromatography (chloroform) to obtain Compound 11 as a white solid.
The analytical value of Compound 11 is shown below.
HRMS (ESI-QTOF MS, positive) Calcd. For C ₄₉ H ₇₂ NO ₁₀ P [M + Na] ⁺ : 888.4786, Found 888.4823
¹ H-NMR (500 MHz, CDCl ₃ ) δ: 7.64 (s, 1H), 7.35 (m, 10H), 4.54 (m, 2H), 4.31 (m, 2H), 4.25 (m, 2H), 3.96 ( m, 2H), 3.52 (s, 3H), 3.47 (m, 1H), 2.47 (m, 2H), 1.62 (s, 6H), 1.26 (s, 6H), 1.40-0.90 (m, 16H), 0.88 -0.78 (m, 15H).

<Example 12>
2-((1-methyl-2-oxo-5-((1r, 2R, 3R, 4S, 5S) -2,3,4,5-tetrahydroxycyclopentyl) -3-((8S, 10R) -4,6 , 8,10-tetramethyldodecyl) -1,2-dihydropyridin-4-yl) oxy) ethyl dihydrogen phosphate (compound (IV))
Compound 11 (0.5 mg, 0.57 μmol) prepared in Example 11 was dissolved in dichloromethane (1.0 mL). The solution was cooled to 0 ° C., TMSBr (0.52 μL, 2.85 μmol) was added dropwise, and the mixture was stirred at room temperature for 6 hours. The reaction solution was cooled to 0 ° C., water (5 μL) was added dropwise, and the mixture was stirred for 5 minutes. The operation of concentrating the reaction solution under reduced pressure, adding toluene and drying azeotropically was repeated three times. The obtained residue was purified by HPLC to give compound (IV) as a white solid.
The analytical value of compound (IV) is shown below.
HRMS (ESI-QTOF MS, negative) Calcd. For C ₂₉ H ₅₂ NO ₁₀ P [MH] ^- : 604.3256, Found 604.3251
¹ H-NMR (700 MHz, CD ₃ OD) δ: 7.74 (s, 1H), 5.18 (m, 4H), 3.70-3.55 (m, 4H), 3.43 (s, 3H), 3.23 (m, 1H) , 1.96 (m, 2H), 1.40-1.05 (m, 16H), 1.00-0.80 (m, 15H).

<Example 13>
<Screening method of TLR4 ligand>
Two types of Ba / F3 cells used as signal reporter cells for searching for ligands of mouse TLR4 or human TLR4 were obtained from the Department of Infectious Genetics, Institute of Medical Science, University of Tokyo. This cell expresses mouse or human TLR4, MD-2, and CD14, and a reporter gene linked to GFP is introduced downstream of the NF-κB promoter (Honda et al. (2012) J. Leukoc Biol. 91 (6): 967-976). When TLR4 is activated in Ba / F3 cells, expression of NF-κB is induced, and GFP existing downstream of the NF-κB promoter is expressed. Therefore, TLR4 activation can be detected by detecting cellular GFP.
Compound (I), Compound (II), Compound (III) and Compound (IV) synthesized in Examples 1, 2, 7 and 12 were dissolved in dimethyl sulfoxide (DMSO) to 2 to 10 mg / mL. Used.

Ba cultured in RPMI 1640 medium containing 10% (v / v) fetal calf serum (FCS), 50 U / mL penicillin, 50 μg / mL streptomycin, 2 mM L-glutamine, 50 μM 2-mercaptoethanol and 1 ng / mL interleukin-3. / F3 cells are dispensed into a 96-well plate so that the number of cells per well is 1.0 × 10 ⁵ , and compound (I), compound (II), compound (III) or compound (IV) Were added to a final concentration of 10 μg / mL, 50 μg / mL or 100 μg / mL, respectively, and incubated at 37 ° C. in the presence of 5% CO ₂ for 18 hours. For control, DMSO was added to the Ba / F3 cell culture solution to a final concentration of 2% (v / v) and incubated for 18 hours in the same manner. Further, as a positive control, a DMSO solution of lipid A (manufactured by Sigma) was added to the culture medium of the Ba / F3 cells so that the final concentration of lipid A was 3 ng / mL, and incubated for 18 hours in the same manner.

  Ba / F3 cells added with compound (I), compound (II), compound (III), compound (IV), lipid A, or DMSO are incubated (18 hours), and then centrifuged (1,200 rpm, 2 The culture medium was removed, and the cells were suspended in PBS (FACS buffer) containing 200 μL of 2.5% FCS per well. After this washing operation was repeated twice, the cells were suspended in FACS buffer containing 0.25 μg / mL 7-amino-actinomycin D. The GFP fluorescence intensity of the cells was analyzed by flow cytometry. Flow cytometry was measured with FACSCanto (registered trademark) II (Becton Dickinson), and data was analyzed with FlowJo software (Tree Star).

<Results of flow cytometry (FACS) analysis>
Representative results are shown in FIGS. 1A-1H. 1A to 1D show the results of analysis using mouse TLR4 expressing cells, and FIGS. 1E to 1H show the results of using human TLR4 expressing cells. FIG. 1A is a FACS histogram of cells to which compound (I) was added to a final concentration of 10 μg / mL. FIG. 1B is a FACS histogram of cells to which compound (II) was added to a final concentration of 50 μg / mL. FIG. 1C is a FACS histogram of cells to which compound (II) was added to a final concentration of 100 μg / mL. FIG. 1D is a FACS histogram of cells with 3 ng / mL lipid A added as a positive control. FIG. 1E is a FACS histogram of cells to which compound (I) was added to a final concentration of 50 μg / mL. FIG. 1F is a FACS histogram of cells to which compound (III) was added to a final concentration of 50 μg / mL. FIG. 1G is a FACS histogram of cells to which compound (IV) was added to a final concentration of 50 μg / mL. FIG. 1H is a FACS histogram of cells with 3 ng / mL lipid A added as a positive control. 1A to 1H, the left peak (white) is a control (when DMSO is added).
From the results shown in FIGS. 1A to 1C, it can be seen that GFP was expressed in mouse TLR4-expressing Ba / F3 cells by the addition of compound (I) and compound (II). From the results shown in FIGS. 1F to 1G, it can be seen that GFP was expressed in human TLR4-expressing Ba / F3 cells by the addition of compound (III) and compound (IV). Therefore, it was shown that Compound (I) and Compound (II) have an action of activating mouse TLR4, and Compound (I), Compound (III) and Compound (IV) have an action of activating human TLR4.

<Example 14>
<Toxicity test>
The Ba / F3 cells in which expression of GFP was detected in Example 13 were evaluated for toxicity of Compound (I), Compound (II), Compound (III), Compound (IV), and Lipid A. As a positive control, compound 3 obtained in step 2 of Example 2 was added to Ba / F3 cells instead of compound (I) or (II) and incubated for 18 hours by the method described in Example 13. Toxicity was evaluated.
Toxicity to Ba / F3 cells was analyzed by flow cytometry using staining with 7-amino-actinomycin D labeled with PerCP-Cy5.5 as an index. Representative results are shown in FIGS. 2A-2I. 2A to 2E show the results of analysis using mouse TLR4 expressing cells, and FIGS. 2F to 2I show the results of using human TLR4 expressing cells. FIG. 2A is a FACS development view of cells to which compound (I) was added to a final concentration of 10 μg / mL. FIG. 2B is a FACS development view of cells to which compound (II) was added to a final concentration of 100 μg / mL. FIG. 2C is a FACS development view of cells to which Compound 3 obtained in Step 2 of Example 2 was added to a final concentration of 50 μg / mL as a positive control. FIG. 2D is a FACS development view of cells to which 3 ng / mL lipid A was added. FIG. 2E is a control (when DMSO is added). FIG. 2F is a FACS development view of cells to which compound (I) was added to a final concentration of 50 μg / mL. FIG. 2G is a FACS development view of cells to which compound (III) was added to a final concentration of 50 μg / mL. FIG. 2H is a FACS development view of cells to which compound (IV) was added to a final concentration of 50 μg / mL. FIG. 2I is a FACS development view of cells to which 3 ng / mL lipid A was added. In FIG. 2A to FIG. 2I, the numbers shown in the figures represent the percentage of living cells (7-amino-actinomycin D negative). The ratio of viable cells in mouse TLR4 expressing cells is 99.3% (FIG. 2A) when compound (I) is added to a final concentration of 10 μg / mL, and final concentration of 100 μg / mL of compound (II). And 99.4% (FIG. 2B). In addition, the proportion of viable cells when 3 ng / mL lipid A was added was 98.9% (FIG. 2D), and the proportion of viable cells in the control was 99.6% (FIG. 2E). The ratio of living cells in human TLR4-expressing cells is 99.9% (FIG. 2F) when compound (I) is added to a final concentration of 50 μg / mL, and compound (III) has a final concentration of 50 μg / mL. 97.0% (FIG. 2G), and 96.0% (FIG. 2H) when compound (IV) was added to a final concentration of 50 μg / mL. In addition, the ratio of viable cells when 3 ng / mL lipid A was added was 99.5% (FIG. 2I).
The results shown in FIGS. 2A to 2I indicate that the toxicity of compound (I), compound (II), compound (FNC-N-P01) and compound (IV) is low.

Claims (8)

The following general formula (1)
(In the above formula, X represents CH ₂ , CH (OH) or an oxygen atom,
n represents an integer of 0 or 1,
R ¹ represents a hydrogen atom or a linear or branched alkyl having 1 to 6 carbon atoms,
R ² represents a hydrogen atom, —CH ₂ CH ₂ OH, or a linear or branched alkyl group having 1 to 6 carbon atoms having an acidic group , wherein the acidic group is —S (O) (OH) ₂ Group, -OPO (OH) ₂ group, -OP (O) (OBn) ₂ group (Bn represents benzyl group), or -COOH, which may be an ion,
R ³ is the following formula (2)
(In the above formula, the wavy line indicates the bond to the 3- position of the pyridone ring) or the following formula (3)
(Wherein the wavy line represents a bond to the 3- position of the pyridone ring),
R ⁴ , R ⁵ , R ⁶ , and R ⁷ are the same or different and may have a hydroxyl group, an optionally substituted —O-acyl group having 1 to 6 carbon atoms, or a substituent. Represents a good linear or branched alkoxy group having 1 to 6 carbon atoms, or —O-benzyl group, or R ⁴ and R ⁵ , R ⁵ and R ⁶ , or R ⁶ and R ⁷ together A compound represented by a cyclic acetal group (in the general formula (1), n is 0, R ¹ is a methyl group, R ² is a hydrogen atom, R ³ Is the following formula (3-1)
(I) a compound in which R ⁴ , R ⁵ , R ⁶ , and R ⁷ are hydroxyl groups , (ii) R ⁴ and R ⁷ are hydroxyl groups, and R ⁵ and R ⁶ are methoxy groups. a compound, and (iii) R ⁴ and R ⁷ are methoxy methoxy group, excluding the compounds R ⁵ and R ⁶ are methoxy group), or a pharmaceutically acceptable salt thereof. Under following formula (I)
Or the following formula (II)
Or the following formula (III)
Or the following formula (IV)
Or the following formula
Compound represented by. A pharmaceutical composition comprising the compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof. A toll-like receptor 4 activator comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient. An immunostimulant comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient. A preventive or therapeutic agent for a disease associated with Toll-like receptor 4, characterized by comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient. An agent for preventing or treating infection, comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient.   The pharmaceutical composition according to claim 3, which is a composition for enhancing the immunogenicity of an antigen.