Novel aminoalkyl glucosaminide 4-phosphate derivatives

Abstract

The present invention provides a novel compound, which has a TLR4 activating effect and is usable as an immune activator or adjuvant in vaccine or allergen immunotherapy, or a pharmaceutically acceptable salt thereof.　A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof. [In formula (I), X, Y, Z and n are each as defined in the description.]

Description

Technical Field

[0001] The present invention relates to novel compounds having an immunostimulatory effect. In particular, it relates to novel aminoalkylglucosamine 4-phosphate derivatives that enhance the efficacy of vaccines or allergen immunotherapy.

Background Art

[0002] Attenuated viruses and inactivated viruses have been used for infectious disease vaccines. However, from the viewpoints of standardization and safety, vaccines with higher antigen purity and fewer safety issues in vaccine preparations have been developed. Along with this, however, a new problem of reduced immunogenicity has arisen, and adjuvants have come to be added for the purpose of enhancing the efficacy of vaccines (Non-Patent Document 1).

[0003] The endotoxin (endotoxin) of the outer membrane of Gram-negative bacterial cell walls, discovered in 1892, is now recognized as lipopolysaccharide (LPS) and is known as a substance that induces shock symptoms and systemic inflammatory reactions. On the other hand, it is also known that a small amount of endotoxin exhibits a therapeutic effect in some diseases. In vaccine research, the possibility that endotoxin can enhance the vaccine effect has been known since the 1800s (Non-Patent Document 2).

[0004] The existence of receptors that recognize pathogen-associated molecular patterns (PAMPs) was proposed in 1989 (Non-Patent Document 3). This concept was supported by the discovery of mammalian Toll-like receptors (TLRs), and in 1998, TLR4, which is the receptor for LPS, was discovered (Non-Patent Document 4).

[0005] Lipid A was isolated as a constituent of LPS, and it was discovered that Lipid A is important for activating intracellular signal transduction via TLR4 (Non-Patent Documents 5 and 6). Monophosphoryl Lipid A (MPLA) was screened from the LPS fraction of Salmonella typhimurium in 1982 (Non-Patent Document 7), developed as an adjuvant for injectable vaccines, and is used in cervical cancer prevention vaccines, hepatitis B vaccines, etc. In these vaccine preparations, it has been shown that MPLA exhibits an immune activation effect and improves the drug efficacy by enhancing the production of virus antigen-specific IgG in the blood. Also, the usefulness of MPLA in allergen immunotherapy (AIT) has been confirmed, and it has been shown to suppress allergic symptoms by inducing allergen-specific IgG in a short period (Non-Patent Document 8).

[0006] In many human pathogens, an infection route through the mucosa is known, and secretory IgA present on the mucosal surface is important for mucosal infection defense against viruses and bacteria (Non-Patent Document 9). However, general injectable infectious disease vaccines cannot efficiently induce mucosal IgA. On the other hand, since mucosal IgA is efficiently induced in mucosal immunity, the development of mucosal vaccines has been actively carried out (Non-Patent Document 10). However, there are technical hurdles in clinical application, and only a few vaccines have been marketed. The importance of adjuvants is cited as one of the technologies for clinical application (Non-Patent Document 11).

[0007] AIT was discovered as a treatment method for alleviating allergic symptoms by subcutaneously administering allergens that cause allergic rhinitis [subcutaneous immunotherapy (SCIT)] (Non-Patent Document 12). As the mechanism of AIT, it is assumed that the induced allergen-specific IgG, especially IgG4, captures the allergen, thereby competitively inhibiting the binding of the allergen to IgE on effector cells such as mast cells (Non-Patent Documents 13-15). In particular, from the clinical results showing that the symptoms of allergic rhinitis caused by cat antigen were improved by administering an allergen-specific IgG4 preparation against cat antigen (Non-Patent Document 16), it is strongly suggested that allergen-specific IgG4 induced by AIT is the main mechanism for the onset of the therapeutic effect of AIT drugs.

[0008] In SCIT, the need to continue subcutaneous administration about once a week for usually 3 to 5 years and the risk of inducing severe systemic allergic reactions have been pointed out, which has become an issue for widespread use (Non-Patent Document 17). Therefore, sublingual immunotherapy (SLIT) has been investigated as an AIT with higher safety, and a sublingual administration preparation was approved by the US Food and Drug Administration (FDA) in 2011. Subsequently, due to its high convenience in addition to safety, SLIT for various allergens has been rapidly spreading. On the other hand, since the treatment period by SLIT is also as long as 3 to 5 years, there are unmet needs for the onset of therapeutic effects in a shorter period and high drug efficacy.

[0009] For food allergies, since subcutaneous immunotherapy (SCIT) is not performed due to particularly high safety concerns, oral immunotherapy (OIT) is being considered as an alternative. Multiple comparative studies between OIT and sublingual immunotherapy (SLIT) using peanut antigens have also been conducted, and it is generally recognized that OIT is excellent in terms of efficacy, and SLIT is excellent in terms of safety. However, there is an unmet need for higher safety and higher efficacy, and the development of a mucosal immunostimulant (adjuvant) for SLIT formulations is expected as one of them (Non-Patent Document 18).

[0010] As a lipid A-related compound, CRX-527 is known (Patent Document 1).

Prior Art Documents

Patent Documents

[0011]

Patent Document 1

Non-Patent Documents

[0012]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Non-Patent Document 7

Non-Patent Document 8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 13

Non-Patent Document 14

Non-Patent Document 15

Non-Patent Document 16

Non-Patent Document 17

Non-Patent Document 18

Summary of the Invention

Problems to be Solved by the Invention

[0013] The present invention provides a novel compound having a TLR4 activation effect and usable as an immunostimulant or adjuvant in vaccine or allergen immunotherapy, or a pharmaceutically acceptable salt thereof.

Means for Solving the Problems

[0014] The present invention relates to the following (1) to (15).

[0015] (1) General formula (I)

[0016]

Chemical formula

[0017] [In formula (I), X represents an oxygen atom or CH2, Y represents CH2 or C=O, Z represents a halogen atom or OR 1 and R 1 represents a hydrogen atom or the following formula (II):

[0018]

Chemical formula

[0019] [In formula (II), R 2 represents a hydrogen atom or a carboxy group, R 3 represents a hydrogen atom, a hydroxy group, or an acetylamino group, R 4 represents a hydrogen atom or the following formula (III):

[0020]

Chemical formula

[0021] R 5 represents a hydrogen atom or a phosphate group, R 6 represents a hydroxymethyl group, a methyl phosphate group, a carboxy group, or a (1S)-1,2-dihydroxyethyl group] n represents 0 or 1] and a pharmaceutically acceptable salt thereof. (However, a compound in which X represents an oxygen atom, n represents 0, Z represents OR 1 and R 1 represents a hydrogen atom is excluded.)

[0022] (2) In the above formula (I), X represents an oxygen atom or CH2, Y represents C=O, Z represents the following formula (IV):

[0023]

Chemical formula

[0024] [In formula (IV), R 7 represents a hydrogen atom or the one represented by the above formula (III)] n represents 0 or 1, the compound according to (1) or a pharmaceutically acceptable salt thereof. (Provided that the compound in which X represents CH2, n represents 1, and R 7 represents the one represented by the above formula (III) is excluded.)

[0025] (3) In the above formula (I), X represents an oxygen atom, Y represents C=O, Z represents the following formula (V),

[0026] [Chemical formula]

[0027] [In formula (V), R 8 represents a hydroxy group or an acetylamino group, R 9 represents a hydrogen atom or a phosphate group, R 10 represents a hydroxymethyl group, a methyl phosphate group, or a carboxy group] n represents 0, the compound according to (1) or a pharmaceutically acceptable salt thereof.

[0028] (4) Any one compound selected from the following group or a pharmaceutically acceptable salt thereof. (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid, (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-{[3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}propanoic acid, (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-D-glucopyranuronosyl-4-O-phosphono-β-D-glucopyranosyl}oxy)propanoic acid, 6,10-anhydro-8-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-11-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-9-O-phosphono-D-erythro-L-galacto-undecanoate, and 5,9-anhydro-7-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-10-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-8-O-phosphono-D-erythro-L-galacto-decanoic acid.

[0029] (5) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid or a pharmaceutically acceptable salt thereof.

[0030] (6) A pharmaceutical composition comprising the compound according to any one of (1) to (5) or a pharmaceutically acceptable salt thereof.

[0031] (7) A pharmaceutical composition comprising the compound according to any one of (1) to (5) or a pharmaceutically acceptable salt thereof and an antigen.

[0032] (8) A pharmaceutical composition, characterized in that the compound according to any one of (1) to (5) or a pharmaceutically acceptable salt thereof and an antigen are administered in combination simultaneously or at different times.

[0033] (9) The pharmaceutical composition according to (7) or (8), wherein the antigen is one or more selected from the group consisting of attenuated viruses, inactivated viruses, or recombinant proteins of viruses such as influenza virus, adenovirus, rubella virus, mumps virus, RSV, enterovirus, rotavirus, norovirus, or coronavirus, Cryptomeria japonica pollen, Japanese cypress pollen, birch pollen, mugwort pollen, giant hogweed pollen, Japanese lawngrass pollen, Zoysia japonica pollen, spinach pollen, Japanese red pine pollen, dwarf pine pollen, Japanese black pine pollen, chrysanthemum pollen, mugwort pollen, thyme pollen, Bermuda Grass pollen, Kentucky Grass pollen, Meadow Fescue Grass pollen, Orchard Grass pollen, Redtop Grass pollen, Perennial Ryegrass pollen, Sweet Vernal Grass pollen, sumac pollen, mites, cat hair, chicken eggs, milk, peanuts, wheat flour, and buckwheat.

[0034] (10) A pharmaceutical composition according to any one of (6) to (9) for the prevention or treatment of viral infections, allergic diseases, bacterial infections and bacterial-derived toxins, cancer, or intracellular parasitic protozoa.

[0035] (11) A pharmaceutical composition according to any one of (6) to (9) for the prevention or treatment of influenza virus, coronavirus, RSV, norovirus, or rotavirus infections.

[0036] (12) A pharmaceutical composition according to any one of (6) to (9) for the prevention or treatment of allergic diseases caused by Cryptomeria japonica pollen, Chamaecyparis obtusa pollen, horse chestnut pollen, ragweed pollen, Ambrosia artemisiifolia pollen, Phalaris arundinacea pollen, Zoysia japonica pollen, spinach pollen, dwarf pine pollen, dwarf chestnut pollen, Japanese red pine pollen, chrysanthemum pollen, mugwort pollen, timothy pollen, Bermuda grass pollen, Kentucky bluegrass pollen, meadow fescue pollen, orchard grass pollen, redtop grass pollen, perennial ryegrass pollen, sweet vernal grass pollen, Chenopodium album [White Goosefoot, Lamb’s quarters] pollen, mites, cat hair, chicken eggs, milk, peanuts, wheat flour, or buckwheat.

[0037] (13) A TLR4 activator containing the compound according to any one of (1) to (5) or a pharmaceutically acceptable salt thereof.

[0038] (14) An immunostimulant containing the compound according to any one of (1) to (5) or a pharmaceutically acceptable salt thereof.

[0039] (15) The immunostimulant of (14) which is a vaccine adjuvant.

Advantages of the Invention

[0040] The aminoalkylglucosaminide 4-phosphate derivative of the present invention or a pharmaceutically acceptable salt thereof has a TLR4 activating effect and is effective for the prevention or treatment of viral infections, allergic diseases, bacterial infections and bacterial-derived toxins, cancer, or diseases caused by intracellular parasitic protozoa.

Brief Description of Drawings

[0041]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Figure 13

Figure 14

Figure 15

Figure 16

Figure 17

Figure 18

Figure 19

Figure 20

Figure 21

Mode for Carrying Out the Invention

[0042] In the present invention, "*" indicates the binding site to a carbon atom or an oxygen atom.

[0043] The "wavy line" in formula (II) of the present invention indicates that the substituent is present either in the axial position or the equatorial position.

[0044] In the present invention, the "halogen atom" is, for example, a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom. Preferably, it is a fluorine atom.

[0045] Next, preferred substituents in general formula (I) will be described.

[0046] X is preferably an oxygen atom.

[0047] Y is preferably C=O.

[0048] Z is preferably the following (VI) or (VII).

[0049]

Chemical formula

[0050] n is preferably 1.

[0051] A preferred combination of X, Y, Z, and n is that X is an oxygen atom, Y is C=O, Z is the above (VI), and n is 1.

[0052] Another preferred combination of X, Y, Z and n is that X is an oxygen atom, Y is C=O, Z is the above (VII), and n is 0.

[0053] A preferred compound of the present invention is (3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid or a pharmaceutically acceptable salt thereof.

[0054] A preferred compound of the present invention is (2S)-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-3-{[3-deoxy-α-D-manno-octa-2-ulopyranosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranosyl-(2→6)-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}propanoic acid or a pharmaceutically acceptable salt thereof.

[0055] A preferred compound of the present invention is (2S)-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-3-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-D-glucopyranuronosyl-4-O-phosphono-β-D-glucopyranosyl}oxy)propanoic acid or a pharmaceutically acceptable salt thereof.

[0056] A preferred compound of the present invention is 6,10-anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-11-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-9-O-phosphono-D-erythro-L-galacto-undecanoic acid or a pharmaceutically acceptable salt thereof.

[0057] A preferred compound of the present invention is 5,9-anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-10-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-8-O-phosphono-D-erythro-L-galacto-decanoic acid or a pharmaceutically acceptable salt thereof.

[0058] A more preferred compound of the present invention is meglumine (5)(3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoate.

[0059] A more preferred compound of the present invention is sodium (5)(3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoate.

[0060] The compound represented by the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof can be used as either an active ingredient or an additive in pharmaceuticals. Whether it is handled as an active ingredient or an additive depends on the laws of each country.

[0061] The compound represented by the general formula (I) of the present invention can be, if desired, in the form of its pharmaceutically acceptable salt. The pharmaceutically acceptable salt refers to a salt that has no significant toxicity and can be used as a pharmaceutical. The compound represented by the general formula (I) of the present invention can be converted into a salt by reacting it with a base.

[0062] For example, alkali metal salts such as sodium salt, potassium salt, lithium salt; alkaline earth metal salts such as calcium salt, magnesium salt; metal salts such as aluminum salt, iron salt; inorganic salts such as ammonium salt; amine salts such as t-butylamine salt, t-octylamine salt, dibenzylamine salt, morpholine salt, glucosamine salt, phenylglycine alkyl ester salt, ethylenediamine salt, guanidine salt, diethylamine salt, triethylamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, chloroprocaine salt, procaine salt, diethanolamine salt, triethanolamine salt, N-benzylphenethylamine salt, piperazine salt, tetramethylammonium salt, tris(hydroxymethyl)aminomethane salt, meglumine salt and other organic salts can be mentioned. Preferably, they are meglumine salt and sodium salt, and more preferably meglumine salt.

[0063] The compound represented by the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof may take up water molecules and become a hydrate when left in the air or recrystallized, and such hydrates are also included in the compounds or salts of the present invention.

[0064] The compound represented by the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof may absorb certain solvents and form solvates when left in a solvent or recrystallized, and such solvates are also included in the compounds or salts of the present invention.

[0065] In addition, a compound that is converted into the compound represented by the general formula (I), which is the active ingredient of the pharmaceutical composition of the present invention, by reactions with enzymes, gastric acid, etc. under physiological conditions in vivo, that is, a compound that undergoes enzymatic oxidation, reduction, hydrolysis, etc. to change into the compound represented by the general formula (I) or a compound that undergoes hydrolysis, etc. by gastric acid, etc. to change into the compound represented by the general formula (I) is included in the present invention as a "pharmaceutically acceptable prodrug compound".

[0066] Examples of the above prodrugs include, when an amino group is present in the compound represented by the general formula (I), a compound in which the amino group is acylated, alkylated, or phosphorylated (for example, a compound in which the amino group is eicosanoylated, alanylated, pentylaminocarbonylated, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylated, tetrahydrofuranylated, pyrrolidylmethylated, pivaloyloxymethylated, t-butylated, etc.). When a hydroxy group is present in the compound represented by the general formula (I), a compound in which the hydroxy group is acylated, alkylated, phosphorylated, or borated (for example, a compound in which the hydroxy group is acetylated, palmitoylated, propanoylated, pivaloylated, succinylated, fumarylated, alanylated, dimethylaminomethylcarbonylated, etc.). Further, when a carboxy group is present in the compound represented by the general formula (I), a compound in which the carboxy group is esterified or amidated (for example, a compound in which the carboxy group is ethyl esterified, phenyl esterified, carboxymethyl esterified, dimethylaminomethyl esterified, pivaloyloxymethyl esterified, ethoxycarbonyloxyethyl esterified, or methyl amidated, etc.).

[0067] The prodrugs in the present invention can be produced from the compounds represented by the general formula (I) by known methods. Further, the prodrugs in the present invention also include those that change to the compounds represented by the general formula (I) under physiological conditions, as described on pages 163 to 198 of Volume 7, Molecular Design, "Development of Pharmaceuticals" published by Hirokawa Shoten in 1990.

[0068] In the compounds represented by the general formula (I) of the present invention or pharmaceutically acceptable salts thereof, all stereoisomers are included.

[0069] In the compounds represented by the general formula (I) of the present invention or pharmaceutically acceptable salts thereof, these isomers and mixtures of these isomers are all represented by a single formula, namely the general formula (I). Therefore, the present invention includes all of these isomers and mixtures of these isomers in any proportion.

[0070] The compounds represented by the general formula (I) of the present invention or pharmaceutically acceptable salts thereof may also contain non-natural proportions of atomic isotopes in one or more of the atoms constituting such compounds. Examples of atomic isotopes include, for example, deuterium ( 2 H), tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C), etc. Further, the said compounds may be radiolabeled with radioactive isotopes such as, for example, tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C). The radiolabeled compounds are useful as therapeutic or prophylactic agents, research reagents, for example, assay reagents, and diagnostic agents, for example, in vivo imaging diagnostic agents. All isotope variants of the compounds of the present invention, whether radioactive or not, are intended to be included within the scope of the present invention.

[0071] In the present invention, the term "antigen" means a general term for substances that induce an immune response. In particular, a substance containing an antigen that causes an allergic reaction is sometimes referred to as an "allergen". Known antigens or allergens include attenuated viruses, inactivated viruses, proteins recombinantly produced from viral constituent proteins, various pollens, insects, living organisms, and foods. For example, viruses attenuated, inactivated, or recombinantly produced from viral constituent proteins of influenza virus, coronavirus, RSV, norovirus, or rotavirus; viruses attenuated, inactivated, or recombinantly produced from viral constituent proteins of influenza virus, adenovirus, rubella virus, mumps virus, RSV, enterovirus, rotavirus, norovirus, or coronavirus; pollens such as cedar pollen, cypress pollen, birch pollen, ragweed pollen, giant hogweed pollen, canary grass pollen, zoysia pollen, spinach pollen, dwarf pine pollen, dwarf chestnut pollen, red pine pollen, chrysanthemum pollen, mugwort pollen, timothy grass pollen, Bermuda grass pollen, Kentucky grass pollen, meadow fescue grass pollen, orchard grass pollen, redtop grass pollen, perennial ryegrass pollen, sweet vernal grass pollen; pollen of Chenopodium album (white goosefoot, lamb's quarters); mites, cat hair, chicken eggs, milk, peanuts, wheat flour, buckwheat, etc.

[0072] As allergens, more specifically, for example, cedar pollen (Cry j 1, Cry j 2, Cry j 3), cypress pollen (Cha o 1, Cha o 2, Cha o 3), birch pollen (Bet v 1, Bet v 2, Bet v 3, Bet v 4, Bet v 6, Bet v 7, Bet v 8), pigweed pollen (Short Ragweed pollen, Amb a 1, Amb a 2, Amb a 3, Amb a 4, Amb a 5, Amb a 6, Amb a 7, Amb a 8, Amb a 9, Amb a 10, Amb a 11, Amb a 12), giant hogweed pollen, Japanese millet pollen, Japanese lawngrass pollen, spinach pollen, dwarf pine pollen, Japanese hornbeam pollen, Japanese red pine pollen, chrysanthemum pollen, mugwort pollen, timothy grass pollen (Timothy Grass, Phl p 1, Phl p 2, Phl p 4, Phl p 5, Phl p 6, Phl p 7, Phl p 11, Phl p 12, Phl p 13), Bermuda grass pollen (Cyn d 1), Kentucky bluegrass pollen (Poa p 1, Poa p 5, Poa p 9), meadow fescue grass pollen (Fes e 1, Fes e 3, Fes e 4, Fes e 5), orchard grass pollen (Dac g 1, Dac g 2, Dac g 5), redtop grass pollen (Agr a 1), perennial ryegrass pollen (Lol p 1, Lol p 2, Lol p 3, Lol p 5, Lol p 9), sweet vernal grass pollen (Ant o 1), lamb's quarters pollen (Chenopodium album [White Goosefoot, Lamb’s quarters], Che a 1, Che a 2, Che a 3), mites (Der f 1, Der f 2, Der f 3 - 39, Der p 1, Der p 2, Der p 3 - 38), cat hair, chicken eggs (Gal d 1, Gal d 2, Gal d 3, Gal d 4, Gal d 5), cow's milk (Bos d 4, Bos d 5, Bos d 6, Bos d 7, Bos d 8, Bos d 9, Bos d 10, Bos d 11, Bos d 12), peanuts (Ara h 1, Ara h2. It is possible to use Ara h 3, Ara h 4, Ara h 5, Ara h 6, Ara h 7, Ara h 8, Ara h 9, Ara h 10, Ara h 11, Ara h 12, Ara h 13, Ara h 14, Ara h 15, Ara h 16, Ara h 17), wheat flour (Tri a 14, Tri a 15, Tri a 19, Tri a 20, Tri a 21, Tri a 26, Tri a 28, Tri a 29, Tri a 30, Tri a 36), or buckwheat or the like or an extract allergen extract therefrom.

[0073] In the present invention, the "viral infectious disease" refers to a state of being infected with a virus through ingestion, inhalation, being stung by an insect, trauma, sexual contact, etc., and also includes a state of developing a disease resulting from being infected with a virus.

[0074] In the present invention, the "allergy disease" means a systemic or local pathological condition of a living body based on an immune reaction caused by the entry of an allergen into the body. Examples of allergy diseases include allergic rhinitis, food allergy diseases, or atopic dermatitis, etc.

[0075] Reactions due to allergy can be classified into immediate-type allergic reactions and non-immediate-type allergic reactions. An immediate-type allergic reaction refers to a reaction caused by the release of chemical mediators such as histamine and leukotriene from mast cells when an antigen meets in a state where mast cells and IgE antibodies existing in the skin, intestinal mucosa, bronchial mucosa, nasal mucosa, conjunctiva, etc. are bound. A non-immediate-type allergic reaction is a reaction that does not depend on IgE antibodies, and it is suggested that T cells may be involved.

[0076] "Ig" is an abbreviation of Immunoglobulin and refers to an antibody. An antibody is a protein produced and released by B cells and binds to foreign substances such as pathogens that have invaded the body.

[0077] "IgE" is a type of human serum immunoglobulin and is particularly involved in allergic reactions and the like.

[0078] As countermeasures and main treatment methods for allergic diseases, avoidance or removal of the causative antigen, drug therapy with anti-allergic drugs and the like, and allergen immunotherapy (AIT) are known.

[0079] In the present invention, "TLR4" means Toll-like receptor 4, which is a receptor that recognizes molecules characteristic of pathogens. It is known that activation of TLR4 promotes the induction of specific IgG and IgA against antigens.

[0080] "IgG" is a type of human serum immunoglobulin and is involved in detoxification of risk factors and recognition of antigen-antibody complexes by leukocytes and macrophages.

[0081] "IgA" is a type of human serum immunoglobulin and is present in large amounts not only in serum but also in nasal mucus, saliva, breast milk, intestinal fluid, etc., and is involved in mucosal immunity.

[0082] In the present invention, "adjuvant" means a substance that is administered simultaneously or continuously with an antigen or allergen and is used to enhance the immune response against the antigen or allergen.

[0083] In the present invention, "treatment" means recovery, remission, alleviation and / or delay of deterioration of the clinical symptoms of viral infections, allergic diseases, bacterial infections and bacterial-derived toxins, cancer, or diseases caused by intracellular parasitic protozoa in patients suffering from these diseases.

[0084] In the present invention, "prevention" means reducing the incidence of viral infectious diseases, allergic diseases, bacterial infectious diseases and bacterial-derived toxins, cancer, or diseases caused by intracellular parasitic protozoa. Prevention includes reducing the risk of progression of viral infectious diseases, allergic diseases, bacterial infectious diseases and bacterial-derived toxins, cancer, or diseases caused by intracellular parasitic protozoa, or reducing the exacerbation of these diseases. The present invention is effective in preventing the above diseases by inducing the human defensive immune response.

[0085] The compound represented by the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof can be administered in various forms. Examples of the administration forms include oral administration by tablets, capsules, granules, emulsions, pills, powders, syrups (solutions), etc., or parenteral administration by injections (intravenous, intramuscular, subcutaneous or intraperitoneal administration), drip infusions, suppositories (rectal administration), etc. These various preparations can be formulated according to conventional methods using auxiliaries usually used in the pharmaceutical formulation technology field such as excipients, binders, disintegrants, lubricants, flavoring and odor-correcting agents, solubilizing agents, suspending agents, coating agents, etc. for the active ingredient.

[0086] When used as tablets, excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, and silicic acid can be used as carriers; binders such as water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate, and polyvinyl pyrrolidone; disintegrants such as dried starch, sodium alginate, agar powder, laminaran powder, sodium hydrogen carbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, monoglyceride stearate, starch, and lactose; disintegration inhibitors such as sucrose, stearin, cocoa butter, and hydrogenated oil; absorption promoters such as quaternary ammonium salts and sodium lauryl sulfate; humectants such as glycerin and starch; adsorbents such as starch, lactose, kaolin, bentonite, and colloidal silicic acid; and lubricants such as purified talc, stearate, boric acid powder, and polyethylene glycol can be used. Further, tablets with a normal coating, such as sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets, or double tablets and multilayer tablets, can be prepared as needed.

[0087] When used as pills, excipients such as glucose, lactose, cocoa butter, starch, hardened vegetable oil, kaolin, and talc can be used as carriers; binders such as gum arabic powder, tragacanth powder, gelatin, and ethanol; and disintegrants such as laminaran and agar can be used.

[0088] When used as suppositories, those conventionally known in the art can be widely used as carriers, such as polyethylene glycol, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, and semi-synthetic glycerides.

[0089] When used as an injection or sublingual solution, it can be used as a solution, emulsion or suspension. These solutions, emulsions or suspensions are preferably sterilized and isotonic with blood. The solvents used in the production of these solutions, emulsions or suspensions are not particularly limited as long as they can be used as medical diluents. Examples include water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, and the like. In this case, an adequate amount of sodium chloride, glucose or glycerin may be included in the formulation to prepare an isotonic solution, and ordinary solubilizers, buffers, soothing agents, etc. may also be included.

[0090] In addition, the above formulations may, if necessary, include colorants, preservatives, fragrances, flavoring agents, sweeteners, etc., and may further include other pharmaceuticals. The amount of the compound contained in the above formulation is not particularly limited and can be appropriately selected within a wide range. Usually, it is contained in an amount of 0.5 to 70% by weight, preferably 1 to 30% by weight, in the total composition.

[0091] The dosage varies depending on the symptoms, age, etc. of the patient (warm-blooded animal, particularly human). In the case of oral administration, the upper limit is 10 mg (preferably 1 mg) per day, and the lower limit is 0.001 mg for adults. It is desirable to administer it 0 to 3 times a day according to the symptoms.

[0092] Next, a typical production method of the compound represented by the general formula (I) will be described. The compounds of the present invention can be produced by various production methods, and the production methods shown below are examples, and the present invention should not be construed as being limited thereto.

[0093] The compound represented by the general formula (I) of the present invention or a pharmaceutically acceptable salt thereof can be produced by applying various known production methods by utilizing the characteristics based on its basic skeleton or the type of substituents. Known methods include, for example, the methods described in "ORGANIC FUNCTIONAL GROUP PREPARATIONS", 2nd edition, ACADEMIC PRESS, INC., 1989, "Comprehensive Organic Transformations", VCH Publishers Inc., 1989, and the like.

[0094] In this case, depending on the type of functional group present in the compound, it may be effective in terms of production technology to protect the functional group with an appropriate protecting group at the stage of raw material or intermediate, or to replace it with a group that can be easily converted into the functional group.

[0095] Examples of such functional groups include an amino group, a hydroxy group, and a carboxy group, etc. Examples of their protecting groups include the protecting groups described in "Greene’s Protective Groups in Organic Synthesis (4th edition, John Wiley & Sons, Inc., 2006)" by T.W. Greene and P.G. Wuts.

[0096] The protecting group or the group that can be easily converted into the functional group may be appropriately selected and used according to the respective reaction conditions of the production method for compound production.

[0097] According to such a method, after introducing the group and carrying out the reaction, if necessary, by removing the protecting group or converting it into a desired group, the desired compound can be obtained.

[0098] In addition, the prodrug of the compound can be produced by introducing a specific group at the stage of the raw material or intermediate, or by performing a reaction using the obtained compound, in the same manner as the above protecting group. The reaction for producing the prodrug can be carried out by applying methods known to those skilled in the art, such as ordinary esterification, amidation, dehydration, hydrogenation, etc.

[0099] Hereinafter, in order to indicate the compound, the numbers of the compounds shown in each reaction formula are used. That is, it is referred to as "(1)" etc. The compounds with other numbers are also described in the same manner. In the following A to C methods, in the formula, X, Y, and n have the same meanings as described above.

[0100] The abbreviations used in this section, the examples, and the tables have the following meanings. Ac: acetyl group, Bn: benzyl group, Boc: tert-butoxycarbonyl group, Cbz: benzyloxycarbonyl group, CDCl3: deuterated chloroform, CD3OD: deuterated methanol, D2O: heavy water, DMSO: dimethyl sulfoxide, TBDPS: tert-butyldiphenylsilyl group, TES: triethylsilyl group, R: (3R)-3-(decanoyloxy)tetradecanoyl, R’: (3R)-3-(decyloxy)tetradecanoyl.

[0101] Method A The compound represented by (1) of the present invention or a pharmaceutically acceptable salt thereof can be produced according to Method A described below.

[0102]

Chemical formula

[0103]

Chemical formula

[0104] (Step A-1) When X is an oxygen atom in this process, for (1a), glycosylation with (2a) using a Lewis acid is carried out under ice-cooling to produce (7a). A suitable starting material for synthesizing (1a) can be prepared from glucosamine hydrochloride using the procedure described in the report by Imoto et al. (Tetrahedron Lett. 1985, 26, 1545-1548), which is allyl 2-deoxy-4,6-O-(1-methylethylidene)-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-α-D-glucopyranoside.

[0105] (Step A-1’) When X is a carbon atom in this process, (7a) is produced through a carbon-carbon bond formation reaction between (3a) and an alkyne (5a) or (6a). By coupling a lithiated alkyne with (3a) under low-temperature conditions to form a C-glycosyl alkyne, followed by sequential reduction of the nitro group, protection of the primary amine, and reduction of the alkyne under heating conditions, a C-glycosyl amino acid is synthesized. When using alkyne (5a), after deprotecting the protecting group of the hydroxy group, the hydroxy group is oxidized with Dess-Martin to an aldehyde, and the aldehyde is oxidized with Pinnick to be converted to a carboxylic acid. When using alkyne (6a), the deprotection of the acetal group and the oxidation reaction are carried out simultaneously with Jones reagent to form a carboxylic acid. Next, the benzyl protecting group is deprotected by hydrogenation, the carboxylic acid is protected with a benzyl group under heating conditions, the hydroxy groups at the 4,6 positions of glucosamine are protected with an acetal, and the hydroxy group at the 3 position is acylated with (4a) to synthesize (7a).

[0106] (Step A-2) This process is for producing (8a) through deprotection from (7a), acylation with (4a), and phosphorylation at the 4-position. The acetal group of (7a) is deprotected with a mixed solvent of acetic acid and water under heating conditions. After removing the Boc protecting group of the primary amine by acid treatment, the primary amine is amidated using (4a). Next, after removing the Troc protecting group by a reduction reaction, the primary amine is amidated using (4a). After temporarily protecting the 6-hydroxy group of glucosamine with a silyl-based protecting group, the 4-hydroxy group is phosphorylated, and then the silyl-based protecting group is deprotected to synthesize (8a).

[0107] (Step A-3) This process is for producing (1) by deprotecting the benzyl protecting group of (8a) through hydrogenation.

[0108] Method B The compound represented by (2) of the present invention or a pharmaceutically acceptable salt thereof can be produced according to Method B described below.

[0109] [Chemical formula]

[0110] The step from (2a) to (2) can be produced in the same manner as Step A-3 of Method A.

[0111] [Chemical formula]

[0112] (Step B-1) This process is for producing (2a) through glycosylation of (8a) obtained by Method A with a KDO (2-keto-3-deoxy-octulosonic acid) unit (2b). The 6-hydroxy group of glucosamine in (8a) is protected with TES, and after performing a coupling reaction with (2b) using a Lewis acid under ice-cooling, the acetal protecting group is deprotected by acid treatment to synthesize (2a).

[0113] Method C The compound represented by (3) of the present invention or a pharmaceutically acceptable salt thereof can be produced according to Production Method C described below.

[0114]

Chemical formula

[0115] The step from (3a) to (3) can be produced in the same manner as Production Method A-3 of Production Method A.

[0116]

Chemical formula

[0117] (Step C-1) This step is a step of producing (3a) by glycosylation of (2a) obtained by Production Method B and the KDO unit (2b). The 4-position hydroxy group of the KDO site of (2a) is protected with TES, and after performing a coupling reaction with (2b) using a Lewis acid under ice cooling, the acetal protecting group is deprotected by acid treatment to synthesize (3a).

[0118] In the above methods A to C, the acetal protecting group is preferably a dimethylacetal group, but may also be a benzylideneacetal group or the like. Further, the acetal protecting groups for the two hydroxy groups may be independent protecting groups for each. The protecting group for the hydroxy group is preferably a benzyl group or a tert-butyldiphenylsilyl group, but may also be a tert-butyldimethylsilyl group, an allyl group, a benzyloxycarbonyl group or the like. The protecting group for the primary amine is preferably a 2,2,2-trichloroethylcarbonyl group or a tert-butyloxycarbonyl group, but may also be an allyloxycarbonyl group, a benzyloxycarbonyl group, a 9-fluorenylmethyloxycarbonyl group or the like. The protecting groups for the phosphoric acid and carboxylic acid are preferably a benzyl group, but may also be an allyl group, a tert-butyl group, a phenyl group or the like. The low temperature conditions are -100 to -20 °C, preferably -80 to -50 °C. Under ice-cooling, it is -20 to 10 °C, preferably -10 to 5 °C. Under heating conditions, it is 35 to 130 °C, preferably 50 °C to 100 °C. The temperature conditions not described are -10 to 100 °C, preferably 15 °C to 35 °C.

Example

[0119] Examples are given below to explain the present invention in more detail. However, the scope of the present invention is not limited thereto, and these are not construed restrictively in any sense. Also, in this specification, reagents, solvents and starting materials not specifically described are readily available from commercial sources.

[0120] Proton nuclear magnetic resonance spectrum ( 1 1H-NMR) was measured using a JEOL 400 MHz, a Varian 400 MHz, or a Varian 500 MHz nuclear magnetic resonance apparatus. The notation of the spectral data is shown as the chemical shift (relative ppm (δ) with tetramethylsilane as the standard substance), the number of protons, the multiplicity of peak splitting (s: singlet; d: doublet; t: triplet; q: quartet; m: multiplet; br: broad, etc.), and the spin coupling constant as the J value (unit: Hz) when it can be specified.

[0121] The mass spectrum (MS m / z) was measured using the electrospray ionization method (ESI).

[0122] Silica gel column chromatography was performed using commercially available packed columns and an automatic fraction collector (Isorela One manufactured by Biotage, EPCLC-W-Prep2XY manufactured by Yamazen, Purif-α2 manufactured by Shoko Science, etc.), and only the multiple solvent species used as the mobile phase were described. Elution was performed under observation by thin layer chromatography (TLC). As the TLC plate, silica gel 60 F 254 or 60 NH2F 254 s, NH2 silica gel 60 F manufactured by Wako Pure Chemical Industries, Ltd., 254 plates or CHROMATOREX NH TLC manufactured by Fuji Silysia Chemical Ltd. were used. As the developing solvent, the mobile phase used in column chromatography was adopted, and as the detection method, a UV detector or a coloring reagent was adopted, respectively.

[0123] (Example 1) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid ammonium

[0124] [Chemical formula]

[0125] (1a) Allyl 3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-deoxy-4,6-O-(1-methylethylidene)-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-α-D-glucopyranoside To a solution of allyl 2-deoxy-4,6-O-(1-methylethylidene)-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-α-D-glucopyranoside (7.70 g) (Tetrahedron Letters 1985, 26(12), 1545 - 1548) in dichloromethane (80 mL) were added (3R)-3-(decanoyloxy)tetradecanoic acid (6.0 g) (Tetrahedron Letters 2006, 47(13), 2087 - 2092), 4-dimethylaminopyridine (55.5 mg), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (3.49 g) at room temperature, and the mixture was stirred at the same temperature overnight. After adding a saturated aqueous sodium hydrogen carbonate solution to the reaction mixture to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (9.51 g).

[0126] (1b) 3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-deoxy-4,6-O-(1-methylethylidene)-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glucopyranose To a solution of the compound (5.47 g) obtained in Example 1(1a) in tetrahydrofuran (50 mL) was added 1,5-cyclooctadienebis(methyldiphenylphosphine)iridium(I) hexafluorophosphate (284 mg) at room temperature, and the mixture was stirred for 1 minute at the same temperature under a hydrogen atmosphere. After further stirring for 2 hours under a nitrogen atmosphere, water (10 mL), pyridine (1.6 mL), and iodine (3.41 g) were added at the same temperature, and the mixture was stirred for 2 hours. After adding a 5% aqueous sodium thiosulfate solution to the reaction mixture to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (1.89 g).

[0127] (1c) Benzyl (3R)-3-[(tert-butoxycarbonyl)amino]-4-[(3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-deoxy-4,6-O-(1-methylethylidene)-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl)oxy]butanoate To a solution of the compound (930 mg) obtained in Example 1(1b) in dichloromethane (10 mL) were added trichloroacetonitrile (1.2 mL) and 1,8-diazabicyclo[5.4.0]undec-7-ene (0.036 mL) at 0 °C, and the mixture was stirred at the same temperature for 1 hour. After concentrating the reaction solution, the residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the product (1.07 g). To a solution of the product in dichloromethane (10 mL) were added benzyl (3R)-3-[(tert-butoxycarbonyl)amino]-4-hydroxybutanoate (500 mg) and molecular sieves 4A, 1 / 16 (300 mg) at room temperature, and the mixture was stirred for 20 minutes. Then, trimethylsilyl trifluoromethanesulfonate (0.021 mL) was added at 0 °C, and the mixture was stirred at the same temperature for 3 hours. After stopping the reaction by adding triethylamine, the molecular sieves were filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (370 mg).

[0128] (1d) Benzyl (3R)-3-amino-4-[(3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl)oxy]butanoate To a solution of the compound (370 mg) obtained in Example 1(1c) in acetic acid (10 mL) was added water (1 mL) at room temperature, and the mixture was stirred at 60 °C for 2 hours. The reaction solution was concentrated to obtain the product (350 mg). The products (310 mg) obtained by the same method were combined, the product (660 mg) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (4 mL) was added at room temperature, and the mixture was stirred at the same temperature for 1 hour. After adding a saturated aqueous sodium hydrogen carbonate solution to the reaction solution to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [ethyl acetate / methanol] to obtain the title compound (470 mg).

[0129] (1e) Benzyl (3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-[(3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl)oxy]butanoate To a solution of the compound (470 mg) obtained in Example 1(1d) in tetrahydrofuran-methanol (1:1, 8 mL) were added (3R)-3-(decanoyloxy)tetradecanoic acid (400 mg) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (420 mg) at room temperature, and the mixture was stirred overnight at the same temperature. After adding 0.5 N hydrochloric acid to the reaction solution to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate / dichloromethane] to obtain the title compound (520 mg).

[0130] (1f) Benzyl (3R)-4-({2-amino-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-deoxy-β-D-glucopyranosyl}oxy)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (520 mg) obtained in Example 1 (1e) in tetrahydrofuran (5 mL) were added acetic acid (10 mL) and zinc powder (520 mg) at room temperature, and the mixture was stirred at the same temperature for 1 hour. The zinc was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [ethyl acetate / methanol] to obtain the title compound (450 mg).

[0131] (1g) Benzyl (3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-[(3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]butanoate To a solution of the compound (450 mg) obtained in Example 1 (1f) in tetrahydrofuran-methanol (1:1, 8 mL) were added (3R)-3-(decanoyloxy)tetradecanoic acid (317 mg) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (330 mg) at room temperature, and the mixture was stirred overnight at the same temperature. After adding 0.5 N hydrochloric acid to the reaction mixture to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (476 mg).

[0132] (1h) Benzyl (3R)-4-[(6-O-[tert-butyl(diphenyl)silyl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (1.90 g) obtained in Example 1(1g) in dichloromethane (20 mL) were added tert-butyldiphenylchlorosilane (410 mg) and imidazole (21 mg) at room temperature, and the mixture was stirred at the same temperature for 2 hours. After adding a saturated aqueous sodium hydrogen carbonate solution to the reaction mixture to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (2.08 g).

[0133] (1i) Benzyl (3R)-4-[(4-O-[bis(benzyloxy)phosphoryl]-6-O-[tert-butyl(diphenyl)silyl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (2.08 g) obtained in Example 1(1h) in dichloromethane (25 mL) were added dibenzyl N,N-diisopropylphosphoramidite (0.626 mL) and 1H-tetrazole (166 mg) at room temperature, and the mixture was stirred at the same temperature for 1 hour. At 0 °C, 3-chloroperbenzoic acid (400 mg) was added, and the mixture was stirred at the same temperature for 15 minutes. After adding a 5% aqueous sodium thiosulfate solution and a saturated aqueous sodium hydrogen carbonate solution to the reaction mixture to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (2.39 g).

[0134] (1j) Benzyl (3R)-4-[(4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (2.39 g) obtained in Example 1(1i) in tetrahydrofuran (20 mL), acetic acid (0.34 mL) and a 1M solution of tetrabutylammonium fluoride in tetrahydrofuran (5.94 mL) were added at room temperature, and the mixture was stirred at room temperature overnight. After adding water to the reaction solution to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (1.90 g).

[0135] (1k) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid bisammonium To a solution of the compound (88.9 mg) obtained in Example 1(1j) in tetrahydrofuran (3 mL), 10% palladium on carbon (60.0 mg) was added at room temperature, and the mixture was stirred at the same temperature for 8 hours under a hydrogen atmosphere. After filtering off the palladium on carbon, the filtrate was concentrated under reduced pressure. To a solution of the residue in tetrahydrofuran (10 mL), a 4% ammonia solution in methanol (0.25 mL) was added at -78 °C, and the mixture was concentrated under reduced pressure at room temperature. The residue was washed with acetonitrile and collected by filtration to obtain the title compound (57.1 mg).

[0136] (Example 2) Ammonium (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoate

[0137] [Chemical formula]

[0138] (2a) Benzyl (3R)-4-({4-O-[Bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(triethylsilyl)-β-D-glucopyranosyl}oxy)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a dichloromethane solution (5 mL) of the compound (394 mg) obtained in Example 1(1j) were added triethylamine (0.12 mL), 4-dimethylaminopyridine (27.2 mg), and trichloroethylsilane (0.049 mL) at room temperature, and the mixture was stirred at the same temperature for 1 hour. The reaction solution was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (384 mg).

[0139] (2b) Benzyl (3R)-4-[(6-O-[1-Benzyl-7,8-di-O-benzyl-3-deoxy-4,5-O-(1-methylethylidene)-α-D-manno-octa-2-ulopyranonosyl]-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (384 mg) obtained in Example 2 (2a) in dichloromethane (4 mL) were added benzyl (3aR,4R,6S,7aR)-4-[(1R)-1,2-bis(benzyloxy)ethyl]-6-fluoro-2,2-dimethyltetrahydro-2H,4H-[1,3]dioxolo[4,5-c]pyran-6-carboxylate (336 mg) (Angewandte Chemie, International Edition 2001, 40, 1475 - 1480) and molecular sieves 5A, 1 / 16 (1.0 g), and the mixture was stirred at the same temperature for 15 minutes. To the reaction solution was added boron trifluoride diethyl ether complex (0.255 mL) at 0 °C, and the mixture was stirred at the same temperature for 20 minutes. After adding triethylamine to stop the reaction, the molecular sieves were filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (267 mg).

[0140] (2c) Benzyl (3R)-4-{[6-O-(1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-oct-2-ulopyranonosyl)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (267 mg) obtained in Example 2 (2b) in dichloromethane (10 mL) were added water (0.48 mL) and trifluoroacetic acid (0.72 mL) at room temperature, and the mixture was stirred at the same temperature for 30 minutes. After adding saturated aqueous sodium hydrogen carbonate solution to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The drying agent was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (237 mg).

[0141] (2d) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid To a solution of the compound (600 mg) obtained in Example 2(2c) in tetrahydrofuran (12 mL) was added 10% palladium on carbon (360 mg) at room temperature, and the mixture was stirred at the same temperature for 7 hours under a hydrogen atmosphere. After filtering off the palladium on carbon, the filtrate was concentrated under reduced pressure to obtain the title compound (420 mg).

[0142] (2e) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid ammonium To a solution of the compound (420 mg) obtained in Example 2(2d) in tetrahydrofuran (20 mL) was added a solution of 4% ammonia in methanol (1.2 mL) at -78°C, and the mixture was concentrated under reduced pressure at room temperature. The residue was washed with acetonitrile and collected by filtration to obtain the title compound (421 mg).

[0143] (Example 3) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-{[3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}butanoic acid ammonium

[0144] [Chem.]

[0145] (3a) Benzyl (3R)-4-[(6-O-[1-Benzyl-7,8-di-O-benzyl-3-deoxy-4-O-(triethylsilyl)-α-D-manno-octa-2-ulopyranonosyl]-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (620 mg) obtained in Example 2(2c) in dichloromethane (7 mL) were added triethylamine (0.38 mL), 4-dimethylaminopyridine (33.5 mg), and trichloroethylsilane (0.232 mL) at room temperature, and the mixture was stirred at the same temperature for 1 hour. The reaction solution was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (606 mg).

[0146] (3b) Benzyl (3R)-4-{[1-Benzyl-7,8-di-O-benzyl-3-deoxy-4,5-O-(1-methylethylidene)-α-D-manno-octa-2-ulopyranonosyl-(2→4)-1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (606 mg) obtained in Example 3 (3a) in dichloromethane (5 mL) were added benzyl (3aR,4R,6S,7aR)-4-[(1R)-1,2-bis(benzyloxy)ethyl]-6-fluoro-2,2-dimethyltetrahydro-2H,4H-[1,3]dioxolo[4,5-c]pyran-6-carboxylate (655 mg) and molecular sieves 5A, 1 / 16 (2.0 g) at room temperature, and the mixture was stirred at the same temperature for 20 minutes. To the reaction solution was added boron trifluoride diethyl ether complex (0.0415 mL) at 0 °C, and the mixture was stirred at the same temperature for 1 hour. After stopping the reaction by adding triethylamine, the molecular sieves were filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (383 mg).

[0147] (3c) Benzyl (3R)-4-{[1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-oct-2-ulopyranonosyl-(2→4)-1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-oct-2-ulopyranonosyl-(2→6)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(canoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}butanoate To a solution of the compound (484 mg) obtained in Example 3 (3b) in dichloromethane (12 mL) were added water (0.24 mL) and trifluoroacetic acid (0.36 mL) at room temperature, and the mixture was stirred at the same temperature for 3 hours. After stopping the reaction by adding saturated aqueous sodium hydrogen carbonate solution, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The drying agent was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (269 mg).

[0148] (3d) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-{[3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}butanoic acid ammonium Using the compound (236 mg) obtained in Example 3(3c), the reaction was carried out in the same manner as in Example 1(1k) to obtain the title compound (151 mg).

[0149] (Example 4) (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)propanoic acid ammonium

[0150] [Chemical formula]

[0151] (4a) Benzyl (2S)-3-({4-O-[Bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(triethylsilyl)-β-D-glucopyranosyl}oxy)-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate Benzyl (2S)-3-[(4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate (1.40 g) (Bioorganic & Medicinal Chemistry Letters 2008,18,5350 - 5354) was used to carry out the reaction in the same manner as in Example 2(2a) to obtain the title compound (1.49 g).

[0152] (4b) Benzyl (2S)-3-[(6-O-[1-benzyl-7,8-di-O-benzyl-3-deoxy-4,5-O-(1-methylethylidene)-α-D-manno-octa-2-ulopyranonosyl]-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate Using the compound (1.49 g) obtained in Example 4(4a), the reaction was carried out in the same manner as in Example 2(2b) to obtain the title compound (1.39 g).

[0153] (4c) Benzyl (2S)-3-{[6-O-(1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate Using the compound (1.39 g) obtained in Example 4(4b), the reaction was carried out in the same manner as in Example 2(2c) to obtain the title compound (1.25 g).

[0154] (4d) (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)propanoic acid ammonium Using the compound (309 mg) obtained in Example 4(4c), the reaction was carried out in the same manner as in Example 1(1k) to obtain the title compound (213 mg).

[0155] (Example 5) (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-{[3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}propanoic acid ammonium

[0156] [Chemical formula]

[0157] (5a) Benzyl (2S)-3-[(6-O-[1-benzyl-7,8-di-O-benzyl-3-deoxy-4-O-(triethylsilyl)-α-D-manno-octa-2-ulopyranonosyl]-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}propanoate Using the compound (619 mg) obtained in Example 4 (4c), the reaction was carried out in the same manner as in Example 3 (3a) to obtain the title compound (614 mg).

[0158] (5b) Benzyl (2S)-3-{[1-benzyl-7,8-di-O-benzyl-3-deoxy-4,5-O-(1-methylethylidene)-α-D-manno-octa-2-ulopyranonosyl-(2→4)-1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate Using the compound (614 mg) obtained in Example 5 (5a), the reaction was carried out in the same manner as in Example 3 (3b) to obtain the title compound (527 mg).

[0159] (5c) Benzyl (2S)-3-{[1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate Using the compound (484 mg) obtained in Example 5 (5b), the reaction was carried out in the same manner as in Example 3 (3c) to obtain the title compound (319 mg).

[0160] 5(5d) (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-{[3-Deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}propanoic acid ammonium Using the compound (264 mg) obtained in Example 5(5c), the reaction was carried out in the same manner as in Example 1(1k) to obtain the title compound (167 mg).

[0161] (Example 6) (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-D-glucopyranuronosyl-4-O-phosphono-β-D-glucopyranosyl}oxy)propanoic acid ammonium

[0162] [Chemical formula]

[0163] (6a) Benzyl (2S)-3-{[6-O-(6-Benzyl-2,3,4-tri-O-benzyl-D-glucopyranuronosyl)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate To a solution of benzyl 2,3,4-tri-O-benzyl-D-glucopyranuronate (170 mg) in dichloromethane (3 mL) were added trichloroacetonitrile (0.31 mL) and 1,8-diazabicyclo[5.4.0]undec-7-ene (0.009 mL) at 0 °C, and the mixture was stirred at the same temperature for 1 hour. After concentrating the reaction mixture, the residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the product (201 mg). To a solution of the above product in dichloromethane (3 mL) were added benzyl (2S)-3-[(4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate (150 mg) and molecular sieves 4A, 1 / 16 (100 mg) at room temperature, and the mixture was stirred for 20 minutes. Then, trimethylsilyl trifluoromethanesulfonate (0.003 mL) was added at 0 °C, and the mixture was stirred at the same temperature for 30 minutes. After adding triethylamine to stop the reaction, the molecular sieves were filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the less polar diastereomer (6a-1, 48.7 mg) and the more polar diastereomer (6a-2, 59.7 mg) of the title compound.

[0164] (6b) (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-D-glucopyranuronosyl-4-O-phosphono-β-D-glucopyranosyl}oxy)propanoic acid ammonium Using the more polar diastereomer (6a-2, 59.7 mg) obtained in Example 6 (6a), the reaction was carried out in the same manner as in Example 1 (1k) to obtain the title compound (29.4 mg).

[0165] (Example 7) Ammonium (2S)-3-{[6-O-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}propanoate

[0166] [Chemical formula]

[0167] (7a) Benzyl (2S)-3-({4-O-[Bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3,4,6-tri-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl)-β-D-glucopyranosyl}oxy)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}propanoate Using 3,4,6-tri-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glucopyranose (250 mg) (Peptide Science (2009), 45th, 179 - 182), the reaction was carried out in the same manner as in Example 6(6a) to obtain the title compound (187 mg).

[0168] (7b) Benzyl (2S)-3-{[6-O-(2-Acetamido-3,4,6-tri-O-benzyl-2-deoxy-β-D-glucopyranosyl)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate To a solution of the compound (187 mg) obtained in Example 7(7a) in acetic acid (4 mL) was added zinc powder (200 mg) at room temperature, and the mixture was stirred at the same temperature for 1 hour. The zinc was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [dichloromethane / methanol] to give the product (90.6 mg). To a solution of the above product in dichloromethane-methanol (1:2, 3 mL) were added triethylamine (0.1 mL) and acetic anhydride (0.5 mL) at room temperature, and the mixture was stirred at room temperature for 15 minutes. The reaction was stopped by adding saturated aqueous sodium hydrogen carbonate solution to the reaction mixture, and then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to give the title compound (57.2 mg).

[0169] (7c) (2S)-3-{[6-O-(2-Acetamido-2-deoxy-β-D-glucopyranosyl)-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoic acid ammonium Using the compound (57.2 mg) obtained in Example 7(7b), the reaction was carried out in the same manner as in Example 1(1k) to give the title compound (28.5 mg).

[0170] (Example 8) Ammonium (2S)-3-({6-O-[2-acetamido-2-deoxy-4-O-(hydroxyphosphinato)-β-D-glucopyranosyl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl}oxy)-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate

[0171] [Chemical formula]

[0172] (8a) Allyl 3-O-benzyl-6-O-[(benzyloxy)carbonyl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-α-D-glucopyranoside To a solution of allyl 3-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-α-D-glucopyranoside (3.78 g) (Journal of Endotoxin Research 1994, 1(3), 149 - 163) in tetrahydrofuran (30 mL) were added pyridine (1.26 mL) and carbobenzoxychloride (1.67 mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction was stopped by adding saturated aqueous sodium hydrogen carbonate solution to the reaction mixture, and then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate / dichloromethane] to obtain the title compound (3.88 g).

[0173] (8b) Allyl 3-O-benzyl-6-O-[(benzyloxy)carbonyl]-4-O-[bis(benzyloxy)phosphoryl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-α-D-glucopyranoside Using the compound (3.88 g) obtained in Example 8 (8a), the reaction was carried out in the same manner as in Example 1 (1i) to obtain the title compound (5.51 g).

[0174] (8c) 3-O-Benzyl-6-O-[(benzyloxy)carbonyl]-4-O-[bis(benzyloxy)phosphoryl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glucopyranose Using the compound (5.51 g) obtained in Example 8 (8b), the reaction was carried out in the same manner as in Example 1 (1b) to obtain the title compound (4.11 g).

[0175] (8d) Benzyl (2S)-3-{[6-O-(3-O-benzyl-6-O-[(benzyloxy)carbonyl]-4-O-[bis(benzyloxy)phosphoryl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl)-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl]oxy}-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate Using the compound (380 mg) obtained in Example 8 (8c), the reaction was carried out in the same manner as in Example 6 (6a) to obtain the title compound (261 mg).

[0176] (8e) Benzyl (2S)-3-[(6-O-{2-acetamido-3-O-benzyl-6-O-[(benzyloxy)carbonyl]-4-O-[bis(benzyloxy)phosphoryl]-2-deoxy-β-D-glucopyranosyl}-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate To a solution of the compound (261 mg) obtained in Example 8 (8d) in acetic acid (4 mL) was added zinc powder (520 mg) at room temperature, and the mixture was stirred at the same temperature for 1 hour. The zinc was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [dichloromethane / methanol] to give the product (131 mg). To a solution of the above product in pyridine (1 mL) was added acetic anhydride (1 mL) at room temperature, and the mixture was stirred at room temperature for 30 minutes. After the reaction solution was concentrated under reduced pressure, the residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to give the title compound (102 mg).

[0177] (8f) (2S)-3-({6-O-[2-Acetamido-2-deoxy-4-O-(hydroxyphosphinato)-β-D-glucopyranosyl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl}oxy)-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoic acid ammonium Using the compound (102 mg) obtained in Example 8 (8e), the reaction was carried out in the same manner as in Example 1 (1k) to give the title compound (62.1 mg).

[0178] (Example 9) (2S)-3-({6-O-[2-Acetamido-2-deoxy-6-O-(hydroxyphosphinato)-β-D-glucopyranosyl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl}oxy)-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoic acid ammonium

[0179]

Chemical formula

[0180] (9a) Allyl 3,4-di-O-benzyl-6-O-[bis(benzyloxy)phosphoryl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-α-D-glucopyranoside Using allyl 3,4-di-O-benzyl-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-α-D-glucopyranoside (3.11 g) (Synlett 2007,1,164-166), the reaction was carried out in the same manner as in Example 1(1i) to obtain the title compound (4.52 g).

[0181] (9b) 3,4-di-O-benzyl-6-O-[bis(benzyloxy)phosphoryl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glucopyranose Using the compound (4.52 g) obtained in Example 9(9a), the reaction was carried out in the same manner as in Example 1(1b) to obtain the title compound (3.34 g).

[0182] (9c) Benzyl (2S)-3-({4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3,4-di-O-benzyl-6-O-[bis(benzyloxy)phosphoryl]-2-deoxy-2-{[(2,2,2-trichloroethoxy)carbonyl]amino}-β-D-glucopyranosyl)-β-D-glucopyranosyl}oxy)-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate Using the compound (360 mg) obtained in Example 9(9b), the reaction was carried out in the same manner as in Example 6(6a) to obtain the title compound (215 mg).

[0183] (9d) Benzyl (2S)-3-[(6-O-{2-Acetamido-3,4-di-O-benzyl-6-O-[bis(benzyloxy)phosphoryl]-2-deoxy-β-D-glucopyranosyl}-4-O-[bis(benzyloxy)phosphoryl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-β-D-glucopyranosyl)oxy]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoate Using the compound (169 mg) obtained in Example 9(9c), the reaction was carried out in the same manner as in Example 8(8e) to obtain the title compound (108 mg).

[0184] (9e) (2S)-3-({6-O-[2-Acetamido-2-deoxy-6-O-(hydroxyphosphinato)-β-D-glucopyranosyl]-3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl}oxy)-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}propanoic acid ammonium Using the compound (70.9 mg) obtained in Example 9(9d), the reaction was carried out in the same manner as in Example 1(1k) to obtain the title compound (43.3 mg).

[0185] (Example 10) 6,10-Anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-9-O-phosphono-D-erythro-L-galacto-undecanoic acid ammonium

[0186] [Chemical formula]

[0187] (10a) tert-Butyl [(3R)-5-{[tert-butyl(diphenyl)silyl]oxy}penta-1-yn-3-yl]carbamate To a solution of tert-butyl [(2R)-4-{[tert-butyl(diphenyl)silyl]oxy}-1-oxobutan-2-yl]carbamate (27.7 g) (Tetrahedron Letters 2007, 48, 7279 - 7282) in methanol (270 mL) were added dimethyl (1-diazo-2-oxopropyl)phosphonate (14.1 mL) and potassium carbonate (17.4 g) at 0 °C, and the mixture was stirred at room temperature overnight. The reaction was quenched by adding saturated aqueous ammonium chloride solution, and the mixture was extracted with n-hexane. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The drying agent was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to give the title compound (23.9 g).

[0188] (10b) 6,10-Anhydro-8,9,11-tri-O-benzyl-3-[(tert-butoxycarbonyl)amino]-1-O-[tert-butyl(diphenyl)silyl]-2,3,4,5,7-pentadeoxy-7-nitro-D-erythro-L-galacto-undec-4-ynitol To a solution of the compound (23.9 g) obtained in Example 10(10a) in tetrahydrofuran (200 mL) was added dropwise a 1.6 M n-hexane solution of n-butyllithium (75 mL) at -78 °C, and the mixture was stirred at the same temperature for 1 hour. To a solution of 1,5-anhydro-3,4,6-tri-O-benzyl-2-deoxy-2-nitro-D-arabino-hex-1-enitol (25.2 g) (European Journal of Organic Chemistry 1998, 8, 1609-1613) in tetrahydrofuran (200 mL) was added dropwise the tetrahydrofuran solution of the organolithium compound prepared above at -78 °C. After stirring at the same temperature for 1 hour, a saturated aqueous ammonium chloride solution was added to the reaction mixture to stop the reaction, and then the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (18.4 g).

[0189] (10c) 7-Amino-6,10-anhydro-8,9,11-tri-O-benzyl-3-[(tert-butoxycarbonyl)amino]-1-O-[tert-butyl(diphenyl)silyl]-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undec-4-ynitol To a solution of the compound (4.47 g) obtained in Example 10(10b) in tetrahydrofuran (25 mL) were added acetic acid (25 mL) and zinc powder (3.30 g) at room temperature, and the mixture was stirred at the same temperature for 7 hours. The zinc was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (2.78 g).

[0190] (10d) 6,10-Anhydro-8,9,11-tri-O-benzyl-3-[(tert-butoxycarbonyl)amino]-1-O-[tert-butyl(diphenyl)silyl]-2,3,4,5,7-pentadeoxy-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undec-4-ynitol To a solution of the compound (2.78 g) obtained in Example 10 (10c) in dichloromethane (30 mL), N,N-diisopropylethylamine (1.11 mL) and 2,2,2-trichloroethyl chloroformate (0.652 mL) were added at room temperature, and the mixture was stirred at the same temperature for 2 hours. After adding a saturated aqueous sodium hydrogen carbonate solution to the reaction mixture to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (2.87 g).

[0191] (10e) 6,10-Anhydro-8,9,11-tri-O-benzyl-3-[(tert-butoxycarbonyl)amino]-1-O-[tert-butyl(diphenyl)silyl]-2,3,4,5,7-pentadeoxy-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undec-4-ynitol To a solution of the compound (2.87 g) obtained in Example 10 (10d) in dimethoxyethane (30 mL), p-toluenesulfonyl hydrazide (4.09 g) was added at room temperature, and 1M aqueous sodium acetate solution (15 mL) was added 6 times at 80 °C at 30-minute intervals. After stirring at the same temperature for 2 hours, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (2.57 g).

[0192] (10f) 6,10-Anhydro-8,9,11-tri-O-benzyl-3-[(tert-butoxycarbonyl)amino]-2,3,4,5,7-pentadeoxy-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undecitol To a solution of the compound (2.57 g) obtained in Example 10 (10e) in tetrahydrofuran (20 mL), acetic acid (0.421 mL) and a 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran (7.35 mL) were added at room temperature, and the mixture was stirred at room temperature overnight. After adding water to the reaction mixture to stop the reaction, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (1.99 g).

[0193] (10g) 6,10-Anhydro-8,9,11-tri-O-benzyl-3-[(tert-butoxycarbonyl)amino]-2,3,4,5,7-pentadeoxy-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undecose To a solution of the compound (1.99 g) obtained in Example 10 (10f) in dichloromethane (30 mL), Dess-Martin periodinane (1.25 g) was added at room temperature, and the mixture was stirred at the same temperature for 3 hours. After adding a saturated aqueous solution of sodium hydrogen carbonate to the reaction mixture to stop the reaction, the mixture was extracted with dichloromethane. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (1.85 g).

[0194] (10h) 6,10-Anhydro-8,9,11-tri-O-benzyl-3-[(tert-butoxycarbonyl)amino]-2,3,4,5,7-pentadeoxy-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undecenoic acid To a solution of the compound (1.85 g) obtained in Example 10 (10 g) in tert-butyl alcohol (20 mL) were added water (4 mL), sodium chlorite (311 mg, 80%), 2-methyl-2-butene (1.5 mL), and sodium dihydrogen phosphate dihydrate (536 mg) at room temperature, and the mixture was stirred at the same temperature for 2 hours. Ethyl acetate was added to the reaction solution for extraction, and the organic layer was washed with saturated brine and then dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (1.64 g).

[0195] (10i) Benzyl 6,10-anhydro-3-[(tert-butoxycarbonyl)amino]-2,3,4,5,7-pentadeoxy-9,11-O-(1-methylethylidene)-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undecenoate To a solution of the compound (1.64 g) obtained in Example 10 (10 h) in tetrahydrofuran (20 mL) was added 10% palladium on carbon (1.6 g) at room temperature, and the mixture was stirred at the same temperature for 8 hours under a hydrogen atmosphere. After filtering off the palladium catalyst, the filtrate was concentrated under reduced pressure to obtain the product. To a solution of the product in N,N-dimethylformamide (10 mL) were added sodium hydrogen carbonate (983 mg) and benzyl bromide (1.18 mL) at room temperature, and the mixture was stirred at 50 °C for 4 hours. Sodium hydrogen carbonate was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [dichloromethane / methanol] to obtain the product. To a solution of the above product in N,N-dimethylformamide (8 mL) were added 2,2-dimethoxypropane (8 mL) and p-toluenesulfonic acid monohydrate (23.6 mg) at room temperature, and the mixture was stirred overnight at the same temperature. After adding triethylamine to stop the reaction, the reaction solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (610 mg).

[0196] (10j) Benzyl 6,10-anhydro-3-[(tert-butoxycarbonyl)amino]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,3,4,5,7-pentadeoxy-9,11-O-(1-methylethylidene)-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undecenoate Using the compound (300 mg) obtained in Example 10(10i), the reaction was carried out in the same manner as in Example 1(1a) to obtain the title compound (380 mg).

[0197] (10k) Benzyl 3-amino-6,10-anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,3,4,5,7-pentadeoxy-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undecenoate Using the compound (380 mg) obtained in Example 10(10j), the reaction was carried out in the same manner as in Example 1(1d) to obtain the title compound (270 mg).

[0198] (10l) Benzyl 6,10-anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-7-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-undecenoate Using the compound (270 mg) obtained in Example 10(10k), the reaction was carried out in the same manner as in Example 1(1e) to obtain the title compound (320 mg).

[0199] (10m) Benzyl 7-amino-6,10-anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecenoate Using the compound (320 mg) obtained in Example 10 (10l), the reaction was carried out in the same manner as in Example 1 (1f) to obtain the title compound (212 mg).

[0200] (10n) Benzyl 6,10-anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecenoate Using the compound (212 mg) obtained in Example 10 (10m), the reaction was carried out in the same manner as in Example 1 (1g) to obtain the title compound (270 mg).

[0201] (10o) Benzyl 6,10-anhydro-11-O-[tert-butyl(diphenyl)silyl]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecenoate Using the compound (1.66 g) obtained in Example 10 (10n), the reaction was carried out in the same manner as in Example 1 (1h) to obtain the title compound (1.93 g).

[0202] (10p) Benzyl 6,10-anhydro-9-O-[bis(benzyloxy)phosphoryl]-11-O-[tert-butyl(diphenyl)silyl]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecenoate Using the compound (1.93 g) obtained in Example 10 (10o), the reaction was carried out in the same manner as in Example 1 (1i) to obtain the title compound (2.15 g).

[0203] (10q) Benzyl 6,10-anhydro-9-O-[bis(benzyloxy)phosphoryl]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecanoate Using the compound (2.15 g) obtained in Example 10 (10p), the reaction was carried out in the same manner as in Example 1 (1j) to obtain the title compound (1.54 g).

[0204] (10r) 6,10-Anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-9-O-phosphono-D-erythro-L-galacto-undecanoic acid ammonium Using the compound (106 mg) obtained in Example 10 (10q), the reaction was carried out in the same manner as in Example 1 (1k) to obtain the title compound (69.0 mg).

[0205] (Example 11) 6,10-Anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-11-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-9-O-phosphono-D-erythro-L-galacto-undecanoic acid ammonium

[0206] [Chemical formula]

[0207] (11a) Benzyl 6,10-anhydro-9-O-[bis(benzyloxy)phosphoryl]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-11-O-(triethylsilyl)-D-erythro-L-galacto-undecenoate Using the compound (1.40 g) obtained in Example 10 (10q), the reaction was carried out in the same manner as in Example 2 (2a) to obtain the title compound (1.49 g).

[0208] (11b) Benzyl 6,10-anhydro-11-O-[1-benzyl-7,8-di-O-benzyl-3-deoxy-4,5-O-(1-methylethylidene)-α-D-manno-octa-2-ulopyranonosyl]-9-O-[bis(benzyloxy)phosphoryl]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecenoate Using the compound (1.49 g) obtained in Example 11 (11a), the reaction was carried out in the same manner as in Example 2 (2b) to obtain the title compound (1.28 g).

[0209] (11c) Benzyl 6,10-anhydro-11-O-(1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-9-O-[bis(benzyloxy)phosphoryl]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecenoate Using the compound (1.24 g) obtained in Example 11 (11b), the reaction was carried out in the same manner as in Example 2 (2c) to obtain the title compound (1.13 g).

[0210] (11d) 6,10 - Anhydro - 8 - O - [(3R)-3 - (decanoyloxy)tetradecanoyl]-3,7 - bis{[(3R)-3 - (decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7 - pentadeoxy - 11 - O - (3 - deoxy - α - D - manno - octa - 2 - ulopyranonosyl)-9 - O - phosphono - D - erythro - L - galacto - undecanoic acid ammonium Using the compound (274 mg) obtained in Example 11(11c), the reaction was carried out in the same manner as in Example 1(1k) to obtain the title compound (192 mg).

[0211] (Example 12) 3 - Deoxy - α - D - manno - octa - 2 - ulopyranonosyl-(2→4)-3 - deoxy - α - D - manno - octa - 2 - ulopyranonosyl-(2→11)-6,10 - anhydro - 8 - O - [(3R)-3 - (decanoyloxy)tetradecanoyl]-3,7 - bis{[(3R)-3 - (decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7 - pentadeoxy - 9 - O - phosphono - D - erythro - L - galacto - undecanoic acid ammonium

[0212] [Chemical formula]

[0213] (12a) Benzyl 6,10 - anhydro - 11 - O - [1 - benzyl - 7,8 - di - O - benzyl - 3 - deoxy - 4 - O - (triethylsilyl)-α - D - manno - octa - 2 - ulopyranonosyl]-9 - O - [bis(benzyloxy)phosphoryl]-8 - O - [(3R)-3 - (decanoyloxy)tetradecanoyl]-3,7 - bis{[(3R)-3 - (decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7 - pentadeoxy - D - erythro - L - galacto - undecanoate Using the compound (562 mg) obtained in Example 11(11c), the reaction was carried out in the same manner as in Example 3(3a) to obtain the title compound (580 mg).

[0214] (12b) Benzyl 1-benzyl-7,8-di-O-benzyl-3-deoxy-4,5-O-(1-methylethylidene)-α-D-manno-octa-2-ulopyranonosyl-(2→4)-1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→11)-6,10-anhydro-9-O-[bis(benzyloxy)phosphoryl]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecenoate Using the compound (580 mg) obtained in Example 12(12a), the reaction was carried out in the same manner as in Example 3(3b) to obtain the title compound (420 mg).

[0215] (12c) Benzyl 1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→11)-6,10-anhydro-9-O-[bis(benzyloxy)phosphoryl]-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-D-erythro-L-galacto-undecenoate Using the compound (420 mg) obtained in Example 12(12b), the reaction was carried out in the same manner as in Example 3(3c) to obtain the title compound (223 mg).

[0216] (12d) 3-Deoxy-α-D-manno-octa-2-ulosopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulosopyranonosyl-(2→11)-6,10-anhydro-8-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-9-O-phosphono-D-erythro-L-galacto-undecanoic acid ammonium salt Using the compound (223 mg) obtained in Example 12(12c), the reaction was carried out in the same manner as in Example 1(1k) to obtain the title compound (141 mg).

[0217] (Example 13) 5,9-Anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium salt

[0218] [Chemical formula]

[0219] (13a) 3,7-Anhydro-5,6,8-tri-O-benzyl-1-[(4S)-3-(tert-butoxycarbonyl)-2,2-dimethyl-1,3-oxazolidin-4-yl]-1,2,4-trideoxy-4-nitro-D-glycero-D-gulo-octa-1-ynitol Using tert-butyl (4S)-4-ethynyl-2,2-dimethyl-1,3-oxazolidine-3-carboxylate (5.50 g) (Tetrahedron 2007,63,8499-8513), the reaction was carried out in the same manner as in Example 10(10b) to obtain the title compound (9.34 g).

[0220] (13b) 4-Amino-3,7-anhydro-5,6,8-tri-O-benzyl-1-[(4S)-3-(tert-butoxycarbonyl)-2,2-dimethyl-1,3-oxazolidin-4-yl]-1,2,4-trideoxy-D-glycero-D-gulo-octa-1-ynitol Using the compound (9.30 g) obtained in Example 13(13a), the reaction was carried out in the same manner as in Example 10(10c) to obtain the title compound (6.40 g).

[0221] (13c) 3,7-Anhydro-5,6,8-tri-O-benzyl-1-[(4S)-3-(tert-butoxycarbonyl)-2,2-dimethyl-1,3-oxazolidin-4-yl]-1,2,4-trideoxy-4-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glycero-D-gulo-octa-1-ynitol Using the compound (6.40 g) obtained in Example 13(13b), the reaction was carried out in the same manner as in Example 10(10d) to obtain the title compound (7.10 g).

[0222] (13d) 3,7-Anhydro-5,6,8-tri-O-benzyl-1-[(4S)-3-(tert-butoxycarbonyl)-2,2-dimethyl-1,3-oxazolidin-4-yl]-1,2,4-trideoxy-4-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-glycero-D-gulo-octitol Using the compound (7.10 g) obtained in Example 13(13c), the reaction was carried out in the same manner as in Example 10(10e) to obtain the title compound (7.10 g).

[0223] (13e) 5,9-Anhydro-7,8,10-tri-O-benzyl-2-[(tert-butoxycarbonyl)amino]-2,3,4,6-tetradeoxy-6-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-talo-decenoic acid To a solution of the compound obtained in Example 13 (13d) (6.70 g) in acetone (120 mL) was added Jones reagent (24 mL) at 0 °C, and the mixture was stirred at room temperature for 1 hour. After adding 2-propanol at 0 °C to stop the reaction, the mixture was extracted with dichloromethane. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [n-hexane / ethyl acetate] to obtain the title compound (4.71 g).

[0224] (13f) Benzyl 5,9-anhydro-2-[(tert-butoxycarbonyl)amino]-2,3,4,6-tetradeoxy-8,10-O-(1-methylethylidene)-6-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-deconate Using the compound obtained in Example 13 (13e) (1.95 g), the reaction was carried out in the same manner as in Example 10 (10i) to obtain the title compound (930 mg).

[0225] (13g) Benzyl 5,9-anhydro-2-[(tert-butoxycarbonyl)amino]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,3,4,6-tetradeoxy-8,10-O-(1-methylethylidene)-6-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-deconate Using the compound obtained in Example 13 (13f) (930 mg), the reaction was carried out in the same manner as in Example 1 (1a) to obtain the title compound (1.23 g).

[0226] (13h) Benzyl 2-amino-5,9-anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,3,4,6-tetradeoxy-6-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-deconate Using the compound obtained in Example 13 (13g) (1.23 g), the reaction was carried out in the same manner as in Example 1 (1d) to obtain the title compound (928 mg).

[0227] (13i) Benzyl 5,9-anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-6-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-decanoate Using the compound (928 mg) obtained in Example 13 (13h), the reaction was carried out in the same manner as in Example 1 (1e) to obtain the title compound (1.24 g).

[0228] (13j) Benzyl 6-amino-5,9-anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-decanoate Using the compound (1.24 g) obtained in Example 13 (13i), the reaction was carried out in the same manner as in Example 1 (1f) to obtain the title compound (960 mg).

[0229] (13k) Benzyl 5,9-anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-decanoate Using the compound (960 mg) obtained in Example 13 (13j), the reaction was carried out in the same manner as in Example 1 (1g) to obtain the title compound (752 mg).

[0230] (13l) Benzyl 5,9-anhydro-10-O-[tert-butyl(diphenyl)silyl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-decanoate Using the compound (2.19 g) obtained in Example 13 (13k), the reaction was carried out in the same manner as in Example 1 (1h) to obtain the title compound (2.28 g).

[0231] (13m) Benzyl 5,9-anhydro-8-O-[bis(benzyloxy)phosphoryl]-10-O-[tert-butyl(diphenyl)silyl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-decenoate Using the compound (2.28 g) obtained in Example 13 (13l), the reaction was carried out in the same manner as in Example 1 (1i) to obtain the title compound (2.62 g).

[0232] (13n) Benzyl 5,9-anhydro-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-decenoate Using the compound (2.62 g) obtained in Example 13 (13m), the reaction was carried out in the same manner as in Example 1 (1j) to obtain the title compound (1.94 g).

[0233] (13o) 5,9-Anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium Using the compound (103 mg) obtained in Example 13 (13n), the reaction was carried out in the same manner as in Example 1 (1k) to obtain the title compound (73.1 mg).

[0234] (Example 14) 5,9-Anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-10-O-(3-deoxy-α-D-manno-oct-2-ulopyranonosyl)-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium salt

[0235] [Chemical formula]

[0236] (14a) Benzyl 5,9-anhydro-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-10-O-(triethylsilyl)-D-erythro-L-galacto-deconate Using the compound (1.40 g) obtained in Example 13(13n), the reaction was carried out in the same manner as in Example 2(2a) to obtain the title compound (1.49 g).

[0237] (14b) Benzyl 5,9-anhydro-10-O-[1-benzyl-7,8-di-O-benzyl-3-deoxy-4,5-O-(1-methylethylidene)-α-D-manno-oct-2-ulopyranonosyl]-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound (1.49 g) obtained in Example 14(14a), the reaction was carried out in the same manner as in Example 2(2b) to obtain the title compound (1.19 g).

[0238] (14c) Benzyl 5,9-anhydro-10-O-(1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound (1.19 g) obtained in Example 14(14b), the reaction was carried out in the same manner as in Example 2(2c) to obtain the title compound (1.10 g).

[0239] (14d) 5,9-Anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-10-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium Using the compound (275 mg) obtained in Example 14(14c), the reaction was carried out in the same manner as in Example 1(1k) to obtain the title compound (196 mg).

[0240] (Example 15) 3-Deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→10)-5,9-anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium

[0241] [Chemical formula]

[0242] (15a) Benzyl 5,9-anhydro-10-O-[1-benzyl-7,8-di-O-benzyl-3-deoxy-4-O-(triethylsilyl)-α-D-manno-octa-2-ulopyranonosyl]-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound (541 mg) obtained in Example 14 (14c), the reaction was carried out in the same manner as in Example 3 (3a) to obtain the title compound (554 mg).

[0243] (15b) Benzyl 1-benzyl-7,8-di-O-benzyl-3-deoxy-4,5-O-(1-methylethylidene)-α-D-manno-octa-2-ulopyranonosyl-(2→4)-1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→10)-5,9-anhydro-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound (554 mg) obtained in Example 15 (15a), the reaction was carried out in the same manner as in Example 3 (3b) to obtain the title compound (422 mg).

[0244] (15c) Benzyl 1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-1-benzyl-7,8-di-O-benzyl-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→10)-5,9-anhydro-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound (422 mg) obtained in Example 15(15b), the reaction was carried out in the same manner as in Example 3(3c) to obtain the title compound (246 mg).

[0245] (15d) 3-Deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→10)-5,9-anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium Using the compound (225 mg) obtained in Example 15(15c), the reaction was carried out in the same manner as in Example 1(1k) to obtain the title compound (145 mg).

[0246] (Example 16) 5,9-Anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6,10-pentadeoxy-10-fluoro-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium

[0247]

Chemical formula

[0248] (16a) Benzyl 5,9-anhydro-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6,10-pentadeoxy-10-fluoro-D-erythro-L-galacto-decenoate To a solution of the compound (142 mg) obtained in Example 13 (13n) in dichloromethane (2 mL) was added bis(2-methoxyethyl)aminosulfur trifluoride (0.05 mL) at 0 °C, and the mixture was stirred at the same temperature for 8 hours. The reaction mixture was quenched by adding saturated aqueous sodium hydrogen carbonate solution, and then extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The drying agent was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography [ethyl acetate / methanol] to give the title compound (90.0 mg).

[0249] (16b) 5,9-Anhydro-7-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2,3,4,6,10-pentadeoxy-10-fluoro-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium Using the compound (90.0 mg) obtained in Example 16 (16a), the reaction was carried out in the same manner as in Example 1 (1k) to give the title compound (56.8 mg).

[0250] (Example 17) 5,9-Anhydro-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium

[0251]

Chemical Structure

[0252] (17a) Benzyl 5,9-anhydro-2-[(tert-butoxycarbonyl)amino]-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2,3,4,6-tetradeoxy-8,10-O-(1-methylethylidene)-6-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-decenoate Using the compound (1.47 g) obtained in Example 13(13f) and (3R)-3-(decyloxy)tetradecanoic acid (928 mg) (Bioorganic & Medicinal Chemistry Letters 2008,18,5350-5354), the reaction was carried out in the same manner as in Example 1(1a) to obtain the title compound (2.51 g).

[0253] (17b) Benzyl 2-amino-5,9-anhydro-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2,3,4,6-tetradeoxy-6-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-decenoate Using the compound (2.05 g) obtained in Example 17(17a), the reaction was carried out in the same manner as in Example 1(1d) to obtain the title compound (444 mg).

[0254] (17c) Benzyl 5,9-anhydro-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2-{[(3R)-3-(decyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-6-{[(2,2,2-trichloroethoxy)carbonyl]amino}-D-erythro-L-galacto-decenoate Using the compound (444 mg) obtained in Example 17(17b) and (3R)-3-(decyloxy)tetradecanoic acid (381 mg), the reaction was carried out in the same manner as in Example 1(1e) to obtain the title compound (590 mg).

[0255] (17d) Benzyl 6-amino-5,9-anhydro-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2-{[(3R)-3-(decyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound obtained in Example 17(17c) (590 mg), the reaction was carried out in the same manner as in Example 1(1f) to obtain the title compound (470 mg).

[0256] (17e) Benzyl 5,9-anhydro-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound obtained in Example 17(17d) (470 mg) and (3R)-3-(decyloxy)tetradecanoic acid (332 mg), the reaction was carried out in the same manner as in Example 1(1g) to obtain the title compound (570 mg).

[0257] (17f) Benzyl 5,9-anhydro-10-O-[(benzyloxy)carbonyl]-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound obtained in Example 17(17e) (570 mg), the reaction was carried out in the same manner as in Example 8(8a) to obtain the title compound (560 mg).

[0258] (17g) Benzyl 5,9-anhydro-10-O-[(benzyloxy)carbonyl]-8-O-[bis(benzyloxy)phosphoryl]-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-D-erythro-L-galacto-deconate Using the compound (190 mg) obtained in Example 17 (17f), the reaction was carried out in the same manner as in Example 1 (1i) to obtain the title compound (221 mg).

[0259] (17h) 5,9-Anhydro-7-O-[(3R)-3-(decyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(decyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-8-O-phosphono-D-erythro-L-galacto-decanoic acid ammonium Using the compound (221 mg) obtained in Example 17 (17g), the reaction was carried out in the same manner as in Example 1 (1k) to obtain the title compound (144 mg).

[0260] (Example 18) Sodium (3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoate To a solution of the compound (104 mg) obtained in Example 2 (2d) in tetrahydrofuran (10 mL), triethylamine (0.033 mL) was added dropwise at room temperature, and then the mixture was concentrated under reduced pressure. DOWEX-50W (1.0 g) was converted to the Na salt with 1N aqueous sodium hydroxide (10 mL) solution, washed with water (10 mL), charged with an aqueous solution (10 mL) of the above compound, and eluted with water (10 mL). The obtained fraction was concentrated under reduced pressure, the above ion exchange operation was repeated twice, dissolved in water, and freeze-dried to obtain the title compound (66.5 mg).

[0261] (Example 19) Potassium (3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoate Using the compound (104 mg) obtained in Example 2(2d) and a 1N potassium hydroxide aqueous solution (10 mL), the reaction was carried out in the same manner as in Example 18 to obtain the title compound (52.1 mg).

[0262] (Example 20) (3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid 4 triethanolamine To a solution of the compound (96 mg) obtained in Example 2(2d) in tetrahydrofuran (3 mL), a solution of triethanolamine (0.04 mL) in tetrahydrofuran (0.4 mL) was added dropwise at room temperature, and then concentrated under reduced pressure. The resulting residue was washed with acetonitrile, the resulting residue was dissolved in water, and freeze-dried to obtain the title compound (116 mg).

[0263] (Example 21) (3R)-3-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid 1 meglumine To a solution of the compound (320 mg) obtained in Example 2(2d) in tetrahydrofuran (5 mL), a solution of meglumine (36 mg) in methanol (5 mL) was added dropwise at room temperature, and then concentrated under reduced pressure. The resulting residue was washed successively with acetonitrile and isopropyl alcohol. The obtained residue was dissolved in water and freeze-dried to obtain the title compound (260 mg).

[0264] (Example 22) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid 2 meglumine Using the compound (300 mg) obtained in Example 2(2d) and meglumine (68 mg), the reaction was carried out in the same manner as in Example 21 to obtain the title compound (336 mg).

[0265] (Example 23) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid 3 meglumine Using the compound (279 mg) obtained in Example 2(2d) and meglumine (95 mg), the reaction was carried out in the same manner as in Example 21 to obtain the title compound (352 mg).

[0266] (Example 24) (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid 4 meglumine Using the compound (780 mg) obtained in Example 2(2d) and meglumine (354 mg), the reaction was carried out in the same manner as in Example 21 to obtain the title compound (1.08 g).

[0267]

Table 1-1

[0268]

Table 1-2

[0269]

Table 1-3

[0270]

Table 1-4

[0271]

Table 1-5

[0272]

Table 1-6

[0273]

Table 1-7

[0274]

Table 1-8

[0275]

Table 1-9

[0276]

Table 1-10

[0277]

Table 1-11

[0278]

Table 1-12

[0279]

Table 1-13

[0280]

Table 1-14

[0281]

Table 1-15

[0282]

Table 1-16

[0283]

Table 2-1

[0284]

Table 2-2

[0285]

Table 2-3

[0286]

Table 2-4

[0287]

Table 2-5

[0288]

Table 2-6

[0289]

Table 2-7

[0290]

Table 2-8

[0291]

Table 2-9

[0292]

Table 2-10

[0293]

Table 2-11

[0294]

Table 2-12

[0295] (Test Example 1) Human TLR4 activation effect Using the compounds described in Examples 1 to 17 and Monophosphoryl Lipid A as Comparative Example A, the human TLR4 activation effect was examined.

[0296] An aqueous Triethanolamine solution in which Triethanolamine was dissolved at 0.5% (v / v) in distilled water for injection was prepared. After dissolving 1 mg of the test drug in 0.98 mL of the aqueous Triethanolamine solution, 20 μL of 1 M HCl was added to adjust the pH to 7.2 - 7.4, and a 1 mg / mL solution was prepared. The dilution series of the test drug was prepared using a medium [DMEM (Nacalai Tesque), 10% FBS (Sigma - Aldrich), 50 U / mL Penicillin - 50 μg / mL Streptomycin (Thermo Fisher Scientific), 1 × HEK - Blue Selection (InvivoGen), 100 μg / mL Normocin (InvivoGen)]. 2 × 10 5 Cells / mL HEK - Blue TMAfter seeding Human TLR4 Cells (InvivoGen), the test drug was administered, and the cells were cultured at 37 °C under 5% CO2 for 24 hours. After culturing, centrifugation was performed at 4 °C and 400 × g for 3 minutes. Recombinant SEAP Protein (InvivoGen) was used for the standard curve. After adding 180 μL of QUANTI-Blue (InvivoGen) to 96 Well TC-Treated Microplates (Corning), 20 μL of the cell culture supernatant and the standard dilution solution were added, and the mixture was allowed to stand at 37 °C for 4 hours. After the reaction, a microplate reader (PerkinElmer, EnSpire) was used to measure the absorbance (wavelength 655 nm), and the SEAP Protein concentration was calculated from the standard curve and used as an index of TLR4 agonist activity. From the linear equation using the two drug concentrations and their measured values that sandwich the drug concentration inducing a reaction of 50% of the maximum reaction in each experiment, the drug concentration inducing a reaction of 50% of the maximum reaction was calculated as EC50 (ng / mL). The results are shown in Table 3.

[0297] [Table 3]

[0298] (Test Example 2) Immunostimulatory effect on ovalbumin (OVA) antigen by sublingual administration (Preparation of test drug) A solution prepared by dissolving 5 μg of Ovalbumin (OVA, Hyglos, Endotoxin Free) and 0.01, 0.1, or 1 μg of the compound described in Example 2 (2e) in 2 μL of distilled water was used as a vaccine preparation, and the immunostimulatory effect on ovalbumin (OVA) antigen by sublingual administration was examined.

[0299] (Sublingual administration test) Mice (BALB / c mice, female, 6 weeks old, Charles River Japan) were fasted and watered from 1 hour before sublingual administration. Thereafter, they were anesthetized with 1-4% vaporized isoflurane (Pfizer). After administering 2 μL of the test drug sublingually, the test drug was maintained sublingually by continuing anesthesia for 10 minutes. After awakening, the second administration was performed 1 hour after the first administration. Fasting and water deprivation were continued for 1 hour after completion of sublingual administration. The above administration was performed for 4 weeks at 1-week intervals. Blood was collected from the tail vein sequentially at 1-week intervals from 2 weeks after administration, and the serum was frozen and stored (-20°C).

[0300] (Measurement of anti-OVA IgG and IgA in blood) 50 μL of OVA (Sigma-Aldrich, 1 μg OVA / mL, PBS) was added to a 96-well Half Area Clear Flat Bottom Polystyrene High Bind Microplate (Corning) and left standing overnight at 4°C, then washed three times with a washing solution (0.05% Tween 20, PBS). 120 μL of ELISA solution (1% BSA, 0.05% Tween 20, PBS) was added, left standing at room temperature for 1 hour, and then washed three times. Serum samples were diluted using the ELISA solution. Anti-OVA Mouse IgG (Chondrex) and Anti-OVA Mouse IgA (Chondrex) were used for the specimens for the standard curve. 50 μL of the samples and the specimens for the standard curve were added, left standing at room temperature for 1 hour, and then washed three times. 50 μL of HRP-labeled anti-mouse IgG (Southern Biotech, diluted 1 / 4000) or IgA (Southern Biotech, diluted 1 / 4000) was added and left standing at room temperature for 1 hour. After washing three times with the washing solution, 50 μL of TMB substrate (SERACARE Life Sciences) was added and left standing for 10 minutes. 50 μL of TMB stop solution (SERACARE Life Sciences) was added to stop the reaction. A microplate reader (PerkinElmer, EnSpire) was used to measure the absorbance at 450 nm. The amounts of anti-OVA IgG and IgA in the blood were calculated from the standard curve. The lower limit of detection was 0.1 and 0.01 μg / mL for anti-OVA IgG and IgA, respectively, and this value was adopted for the values of samples below the detection limit. The results are shown in Figures 1 and 2.

[0301] (Test Example 3) Immunostimulatory effect on various allergens by sublingual administration Using the compound described in Example 2 (2e), the immunostimulatory effect on various allergens by sublingual administration was examined.

[0302] Japanese Cedar Pollen antigen extract was prepared by treating Japanese cedar pollen (Yamizo Pollen Research Group) with a solution containing 0.125 M NaHCO3 and 0.5 M NaCl at 4°C for 24 hours, and then removing insoluble matter by filtration. The concentration of Cry j 1 contained in the extract was measured and diluted to 12.5 μg / mL (10000 JAU / mL). Mite, Ragweed, Timothy, Peanut, and Milk allergens were prepared using Allergen Scratch Extract (Torii Pharmaceutical). Allergen sensitization was performed by administering the allergen (20 μL) into the thigh muscle of mice 1 week before the start of sublingual administration. The allergen scratch extracts of Mite, Ragweed, Timothy, Peanut, and Milk were diluted 10-fold with PBS and used for allergen sensitization. Sublingual administration was started 1 week after sensitization. Various allergens were mixed with distilled water or the compound described in Example 2(2e) dissolved in distilled water at an equal volume immediately before administration to prepare a prototype vaccine formulation. Sublingual administration was performed at 2 μL of the prototype vaccine formulation once a day, 3 - 5 times a week for 12 weeks. Blood was collected from the tail vein 4, 8, and 12 weeks after the start of sublingual administration, and the separated serum was stored at -20°C.

[0303] (Measurement of allergen-specific IgG) To each well of the ELISA plate, 25 μL / well of various allergens were added (cedar pollen, 1 μg Cry j 1 / mL; mite, mugwort, timothy, peanut, milk, diluted 1:1000), and the plate was left standing overnight at 4°C. After 16 - 24 hours, it was washed three times with the washing solution. Then, 100 μL of the ELISA solution was added, left standing at room temperature for 1 hour, and then washed three times. The dilution series of the serum was prepared using the ELISA solution, with the highest concentration being 1 / 128 diluted serum and eight steps prepared by serial two-fold dilution. 25 μL of the sample was added to the plate, left standing at room temperature for 1 hour, and then washed three times. HRP-labeled anti-mouse IgG was diluted 1:10000 with the ELISA solution, 25 μL was added, left standing at room temperature for 1 hour, and then washed three times. 30 μL of the TMB substrate was added, left standing for 10 minutes, then 30 μL of the TMB stop solution was added, and the absorbance at 450 nm was measured. An appropriate OD value was set for each allergen, and the dilution ratio of the serum that reached that OD value was shown as the allergen-specific IgG titer. The results are shown in Figures 3 to 8. In mice sensitized with any of the allergens, in the non-sublingual administration group (Control), no significant increase in allergen-specific IgG titer was observed until 12 weeks later. In any of the allergens, a significantly higher allergen-specific IgG titer was induced in the group co-administered with the compound described in Example 2(2e) sublingually compared to the group administered the allergen alone sublingually.

[0304] (Test Example 4) Immunopotentiating effect on cedar pollen antigen by sublingual administration Using the compound described in Example 24, the immunopotentiating effect on cedar pollen antigen by sublingual administration was examined.

[0305] (Allergen immunotherapy model) Allergen sensitization was performed by administering 50 μL of cedar pollen antigen extract (allergen scratch extract “Tori-i” cedar pollen, Torii Pharmaceutical Co., Ltd.) subcutaneously at the base of the mouse tail 3 and 1 week before the start of sublingual administration. Blood was collected 4 days after the second sensitization, anti-Cry j 1 IgG was measured according to the following section, and the mice were grouped so that the measured values were equal. Sublingual administration (sublingual allergen immunotherapy model) was started 3 days later. As the antigen, cedar pollen antigen (5000 JAU Shidakure [Trademark Registration], Torii Pharmaceutical Co., Ltd.) dissolved in 100 μL of PBS was used. Example 24 was dissolved in injection water (Otsuka Pharmaceutical) and prepared to 5 mg / mL. The antigen and Example 24 were mixed in equal amounts immediately before administration. Mice were fasted and water-deprived starting 1 hour before sublingual administration. Under isoflurane anesthesia, 2 μL of the prototype vaccine preparation was administered sublingually twice at 5-minute intervals, and then the mice were awakened 5 minutes later. The above sublingual administration was performed twice at 1-hour intervals, and then feeding and watering were resumed 1 hour later. This administration was performed 3 times / week. Blood was collected from the tail vein 3 and 4 weeks after the start of sublingual administration. Five weeks later, blood, nasal lavage fluid, and cervical lymph nodes were collected.

[0306] (Measurement of anti-Cry j 1 IgG or IgA) To a 96 Well Micro Plate (Corning, Cat No. 3690), 25 μL of 10 μg / mL ImmunoPure Streptavidin (Thermo Fisher, Cat No. 21125) was added and left standing overnight at 4°C. After washing 3 times with the washing solution, 100 μL of ELISA solution was added and left standing at room temperature for 1 hour. After washing 3 times, 25 μL of 1 μg / mL biotin-labeled Cry j 1 (BioDynamics Laboratory, Cat No. HBL-BC-1) was added and left standing at room temperature for 1 hour. After washing 3 times, 25 μL of the sample was added and left standing at room temperature for 1 hour. After washing 3 times, 25 μL of HRP-labeled anti-mouse IgG (1 / 8000) or HRP-labeled anti-mouse IgA (1 / 4000) was added and left standing at room temperature for 1 hour. After washing 3 times, 30 μL of TMB Microwell Peroxidase Substrate System was added. After leaving standing at room temperature for 10 minutes, 30 μL of TMB Stop Solution was added, and the absorbance at 450 nm was measured. For the standard sample of serum, positive serum was used and the anti-Cry j 1 IgG or IgA contained therein was set at 1000 Unit / mL. This standard sample was prepared in 6 steps from 100-fold dilution to 4-fold dilution to create a standard curve. The evaluation sample was diluted 1000-fold with ELISA solution and measured, and using the standard curve, the Unit value of anti-Cry j 1 IgG or IgA in the evaluation sample was determined. The results are shown in Figures 9 and 10. For the standard sample of nasal lavage fluid, a pooled sample obtained by mixing equal amounts of nasal lavage fluid collected from 4 mice in the positive group was used, and the anti-Cry j 1 IgA contained therein was set at 1000 Unit / mL. This standard sample was prepared in 12 steps from undiluted solution to 2-fold dilution to create a standard curve. The evaluation sample was diluted 8-fold with ELISA solution and measured, and using the standard curve, the Unit value of anti-Cry j 1 IgA in the evaluation sample was determined. The results are shown in Figure 11. In mice without sublingual administration (No-SLIT group) and mice administered only Japanese cedar pollen antigen sublingually (Japanese Cedar Pollen SLIT group), in mice administered Japanese cedar pollen antigen and Example 24 sublingually (Japanese Cedar Pollen + Compound 24 SLIT group), anti-Cry j 1 IgG (Figure 9) and anti-Cry j 1 IgA (Figure 10) in serum were induced to higher levels. Anti-Cry j 1 IgA in nasal lavage fluid was induced to a high level in the Japanese Cedar Pollen + Compound 24 SLIT group (Figure 11).

[0307] (Measurement of Cry j 1-specific cellular immunity) After collecting cervical lymph nodes, they were pooled for each group and ground in PBS (1% BSA). After passing through a 70 μm strainer and adding 5 mL RPMI / FBS / ps (RPMI 1640, 10% FBS, 100 Unit / mL Penicillin and Streptomycin), centrifugation was performed at 400 g for 3 minutes. The cells were collected with RPMI / FBS / ps, and the cell number was 2 × 5 × 10 6Cells were prepared at cells / mL. After seeding 40 μL / well in a 96 Well U-Plate, 40 μL of RPMI or 40 μL of RPMI / FBS / ps containing 2 × 5 μg / mL Cry j 1 was added, and the cells were cultured at 37°C for 44 hours. IL-10, IFN-γ, and IL-4 contained in the culture supernatant were measured using a Mouse Th1 / Th2 / Th17 Cytokine Kit (BD Cytometric Bead Array). The results are shown in FIGS. 12 to 14. Upon stimulation with Cry j 1, IL-10 was strongly induced in the Japanese Cedar Pollen + Compound 24 SLIT group (FIG. 12). On the other hand, both IFN-γ (FIG. 13) and IL-4 (FIG. 14) were below the detection limit (9.77 pg / mL).

[0308] (Measurement of mast cell degranulation by Cry j 1 and suppression of degranulation by IgG derived from immune serum) Mice (C57BL / 6J, 8 months old, male, Charles River Laboratories Japan, Inc.) were euthanized by exsanguination under isoflurane anesthesia. Ice-cold 10 mL of RPMI / BSA / hep (RPMI 1640, 1 mg / mL BSA, 10 units / mL heparin) was injected intraperitoneally, and the abdomen was gently massaged for about 60 seconds, and the peritoneal lavage fluid was collected. The peritoneal lavage fluids from four mice were pooled, centrifuged at 400 g for 3 minutes, the supernatant was discarded, and the peritoneal cells were resuspended in 1 mL of RPMI / BSA / hep. The above RPMI / BSA / hep was warmed to 37°C, and a Histodenz lysate was prepared by dissolving 0.235 g / mL of Histodenz. 2 mL of the Histodenz lysate was added to a 15 mL tube, and 1 mL of the peritoneal cell suspension was overlaid, and centrifuged at 400 g for 15 minutes. The cell population contained in the intermediate layer was discarded, and the peritoneal mast cells at the bottom of the tube were collected with 15 mL of RPMI / FBS / ps (RPMI 1640, 10% FBS, 100 Unit / mL Penicillin-Streptomycin). After centrifugation at 400 g for 3 minutes, the cells were resuspended in 5 mL of RPMI / FBS / ps, seeded into a 24-well plate at 0.5 - 1 mL / well, and immediately 5 μg / mL of Cryj1-01-F11 mouse monoclonal IgE (mIgE), 5 μg / mL of Cryj1-02-F02 mIgE, or both were added. The cells were cultured at 37°C for 24 hours. 10 mL of Tyrode's solution (Sigma Aldrich, T2145-10X1L) was added to the mast cells, and the cells were centrifuged at 400 g for 3 minutes. The cells were approximately 10 6The cells were suspended in Tyrode's solution to a concentration of [[ID=]], and 10 μL of the suspension was added to a 1.5 mL tube (Eppendorf) and allowed to stand at room temperature until the test substance was added. The test substance (Cry j 1; Hayashibara Biochemical Laboratories, Inc., HBL-C-1) was prepared in Tyrode's solution at twice the final concentration. A 2 × 1% Triton-X100 solution was also prepared in Tyrode's solution. After adding 10 μL of the test substance or Triton-X100 to the cells, the cells were immediately allowed to stand at 37°C. After 10 minutes, the cells were immediately cooled on ice to stop the reaction. After centrifugation at 400 g for 3 minutes, 10 μL of the supernatant was added to 50 μL of 4-Nitrophenyl N-acetyl-b-D-glucosaminide (Sigma Aldrich, beta-N-Acetylglucosaminidase Assay Kit). For the background measurement sample, 4-Nitrophenyl N-acetyl-b-D-glucosaminide with only 10 μL of Tyrode's solution added was used. After allowing the samples to stand at 37°C for 1 hour, the reaction was stopped by adding 100 μL of Sodium Carbonate, and the absorbance at 405 nm was measured using a microplate reader (EnSpire [PerkinElmer]). The amount of degranulation (%) was calculated using the following formula: Degranulation (%) = [OD(Sample) - OD(Background)] / [OD(Triton-X100) - OD(Background)] Mast cells that were not sensitized with IgE or were sensitized with only one of two monoclonal IgEs (Cryj1-01-F11 mIgE or Cryj1-02-F02 mIgE) did not degranulate in response to Cry j 1 (Figure 15). This was thought to be because cross-linking of IgE and FcεRI via Cry j 1 did not occur. On the other hand, mast cells sensitized with the two clones degranulated in a Cry j 1 concentration-dependent manner in response to Cry j 1 stimulation (Figures 15 and 16). IgG was purified from 200 μL of serum from each individual of the allergen immunotherapy model mice according to the protocol of ProteinG HP spin trap (Cytiva, 28-9031-34). The purified IgG was used with Vivaspin 500, 10 kDa MWCO Polyethersulfone (Cytiva, 28-9322-25) to replace the solvent with 50 μL of Tyrode's solution. Cry j 1 (2 × 10 × 200 ng / mL, 1.5 μL of Tyrode's solution) was added to the purified IgG (13.5 μL of Tyrode's solution), and the mixture was allowed to stand at room temperature in the dark for 1 hour. 10 μL of this specimen was used as the test substance and treated with mast cells sensitized with anti-Cry j 1 IgE, and the amount of degranulation was measured (stimulated with 200 ng / mL Cry j 1 at the final concentration). IgG purified from the sera of mice in the Japanese Cedar Pollen + Compound 24 SLIT group suppressed the mast cell degranulation reaction via anti-Cry j 1 IgE at a significantly lower level compared to the No-SLIT group and the Japanese Cedar Pollen SLIT group (Figure 17).

[0309] (Test Example 5) Immunostimulatory effect on SARS-CoV-2 RBD by sublingual administration As the antigen, a solution prepared by dissolving the recombinant RBD (Receptor Binding Domain) protein of the novel coronavirus (Severe Acute Respiratory Syndrome Coronavirus 2: SARS-CoV-2) (Recombinant 2019-nCoV RBD protein, Sino Biological, Cat. No. 40592-V08H) at 1 mg / mL was used. The compound described in Example 2 (2e) was dissolved in a 0.5% aqueous solution of triethanolamine and neutralized with HCl to prepare a solution at 1 mg / mL. The antigen and Example 2 (2e) were mixed in equal amounts immediately before administration. Sublingual administration (2 μL) was performed 2 times / day / 1 week. Blood was collected and nasal lavage fluid was collected 6 weeks after the first administration. Nasal lavage was performed with 200 μL of PBS.

[0310] (Measurement of anti-RBD IgG and IgA in blood) Add 25 μL of 10 μg / mL Streptavidin (Thermo Fisher Scientific, Cat#21125, dissolved in PBS) to the ELISA plate, let it stand overnight at 4°C, and then wash it three times. Add 100 μL of the ELISA solution, let it stand at room temperature for 1 hour, and then wash it three times. Add 25 μL of 0.2 μg / mL recombinant RBD protein (Acro Biosystems, Cat. No. SPD-C82E9) prepared with the ELISA solution, let it stand at room temperature for 1 hour, and then wash it three times. Add 25 μL of serum diluted with the ELISA solution, let it stand at room temperature for 1 hour, and then wash it three times. Add 25 μL of HRP-labeled anti-mouse IgG (diluted 1 / 10000) or HRP-labeled anti-mouse IgA (diluted 1 / 4000) diluted with the ELISA solution, let it stand at room temperature for 1 hour, and then wash it three times. Add 30 μL of the TMB substrate, let it stand for 10 minutes, and then add 30 μL of the TMB stop solution to stop the reaction. Measure the absorbance at 450 nm. A sample obtained by mixing equal amounts of the serum of the mice administered with RBD and Example 2 (2e) was used as the standard serum for measurement. The anti-RBD IgG and IgA contained in the standard serum were each set to 1000 Unit / mL to create a standard curve, and the anti-RBD IgG and IgA in each sample were calculated. The values of the samples below the lower limit of the standard curve were set to 0.78 Unit / mL (anti-RBD IgG) and 15.6 Unit / mL (anti-RBD IgA) at the lower limit of the standard curve.

[0311] (Measurement of anti-RBD IgA in nasal lavage fluid) After immobilizing 0.2 μg / mL recombinant RBD protein (Acro Biosystems, SPD-C82E9) in the same manner as in the previous section, add 25 μL of the nasal lavage fluid diluted 1 / 2 with the ELISA solution. The subsequent treatment was performed in the same manner as in the previous section. The result showed the absorbance at 450 nm.

[0312] (RBD-hACE2 binding inhibitory activity in nasal lavage fluid) 25 μL of 10 μg / mL Streptavidin (Thermo Fisher Scientific, Cat#21125, dissolved in PBS) was added to the ELISA plate, allowed to stand overnight at 4°C, and then washed three times. 100 μL of the ELISA solution was added, allowed to stand at room temperature for 1 hour, and then washed three times. 0.04 μg / mL of recombinant RBD protein (Acro Biosystems, SPD-C82E9) was immobilized and washed three times. 25 μL of nasal lavage fluid was added, allowed to stand at room temperature for 1 hour, and then washed three times. 25 μL of 0.1 μg / mL recombinant hACE2 protein (Acro Biosystems, Cat. No. AC2-H5257) diluted with the ELISA solution was added, allowed to stand at room temperature for 1 hour, and then washed three times. 25 μL of HRP-labeled anti-human IgG1 (CYGNUS TECHNOLOGIES, Cat. No. IM50) diluted 500-fold with the ELISA solution was added, allowed to stand at room temperature for 1 hour, and then washed three times. 30 μL of TMB substrate was added, allowed to stand for 10 minutes, and then 30 μL of TMB stop solution was added. The result showed an absorbance at 450 nm.

[0313] (Results) The results of the above test are shown in FIGS. 18 to 21. When only recombinant RBD protein was administered sublingually, anti-RBD IgG or IgA in the blood was below the detection limit. However, anti-RBD IgG (FIG. 18) and IgA (FIG. 19) in the blood of mice co-administered sublingually with the compound described in Example 2 (2e) were significantly higher. Also, a high level of anti-RBD IgA was induced in the nasal lavage fluid of mice co-administered sublingually with the compound described in Example 2 (2e) and recombinant RBD protein (FIG. 20). Although the interaction between the RBD on the surface of the novel coronavirus and hACE2 of the host cell is important for virus infection, the nasal lavage fluid of mice co-administered sublingually with the compound described in Example 2 (2e) and recombinant RBD protein inhibited the binding of RBD and hACE2 (FIG. 21). From the above results, it was suggested that simultaneous sublingual administration of the compound described in Example 2 (2e) and recombinant RBD protein induces RBD-specific IgA in the nasal cavity and may efficiently inhibit the binding of RBD and hACE2 in the nasal cavity, which is the portal of virus entry.

Claims

1. A compound selected from the following group or a pharmaceutically acceptable salt thereof. (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid, (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-{[3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→4)-3-deoxy-α-D-manno-octa-2-ulopyranonosyl-(2→6)-3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-4-O-phosphono-β-D-glucopyranosyl]oxy}propanoic acid, (2S)-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-3-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-D-glucopyranuronosyl-4-O-phosphono-β-D-glucopyranosyl}oxy)propanoic acid, 6,10-Anhydro-8-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-3,7-bis{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2,3,4,5,7-pentadeoxy-11-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-9-O-phosphono-D-erythro-L-galacto-undecanoic acid, and 5,9-Anhydro-7-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2,6-bis{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2,3,4,6-tetradeoxy-10-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-8-O-phosphono-D-erythro-L-galacto-decanoic acid.

2. (3R)-3-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-4-({3-O-[(3R)-3-(Decanoyloxy)tetradecanoyl]-2-{[(3R)-3-(Decanoyloxy)tetradecanoyl]amino}-2-deoxy-6-O-(3-deoxy-α-D-manno-octa-2-ulopyranonosyl)-4-O-phosphono-β-D-glucopyranosyl}oxy)butanoic acid or a pharmaceutically acceptable salt thereof. **Claim 3** A pharmaceutical composition comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. **Claim 4** A pharmaceutical composition comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, and an antigen. **Claim 5** A pharmaceutical composition for administration of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof and an antigen simultaneously or at different times in combination. **Claim 6** The pharmaceutical composition according to claim 4 or 5, wherein the antigen is one or more selected from the group consisting of an attenuated virus, an inactivated virus, or a recombinant protein of a virus component protein of influenza virus, adenovirus, rubella virus, mumps virus, RSV, enterovirus, rotavirus, norovirus, or coronavirus, or cedar pollen, hinoki pollen, birch pollen, hogweed pollen, giant hogweed pollen, Japanese stiltgrass pollen, Japanese mugwort pollen, spinach pollen, Japanese cypress pollen, dwarf pine pollen, Japanese red pine pollen, chrysanthemum pollen, mugwort pollen, thyme pollen, Bermuda Grass pollen, Kentucky Grass pollen, Meadow Fescue Grass pollen, Orchard Grass pollen, Redtop Grass pollen, Perennial Ryegrass pollen, Sweet Vernal Grass pollen, sumac pollen, mite, cat hair, chicken egg, milk, peanut, wheat flour, or buckwheat extract. **Claim 7** The pharmaceutical composition according to any one of claims 3 to 6 for the prevention or treatment of viral infections, allergic diseases, bacterial infections and bacterial-derived toxins, cancer, or intracellular parasitic protozoa. **Claim 8** The pharmaceutical composition according to any one of claims 3 to 6 for the prevention or treatment of influenza virus, coronavirus, RSV, norovirus, or rotavirus infection.

9. The pharmaceutical composition according to any one of claims 3 to 6 for the prevention or treatment of allergic diseases caused by Cryptomeria japonica pollen, Chamaecyparis obtusa pollen, Betula pollen, ragweed pollen, Ambrosia artemisiifolia pollen, Phalaris arundinacea pollen, Zoysia matrella pollen, Spinacia oleracea pollen, Pinus densiflora pollen, Scutellaria baicalensis Georgi pollen, Pinus densiflora pollen, Chrysanthemum morifolium Ramat pollen, Artemisia princeps Pamp pollen, Timothy grass pollen, Bermuda grass pollen, Kentucky grass pollen, Meadow fescue grass pollen, Orchard grass pollen, Redtop grass pollen, Perennial ryegrass pollen, Sweet vernal grass pollen, Chenopodium album (White Goosefoot, Lamb’s quarters) pollen, mites, cat hair, chicken eggs, milk, peanuts, wheat flour, or buckwheat.

10. A TLR4 activator containing the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to any one of claims 3 to 6.

11. An immunostimulant containing the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to any one of claims 3 to 6.

12. The immunostimulant of claim 11 which is a vaccine adjuvant.

Images