JPH0233719B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel glucosamine derivatives with excellent biological activity. More specifically, the present invention provides a general formula that has an excellent immune function enhancing effect. [In the formula, R is a lower alkyl group, R ¹ is a carbon number of 3
~41 alkyl or alkenyl group, formula (In the formula, R ¹¹ represents an alkylene group having 2 to 9 carbon atoms.) or a group represented by the formula In the group represented by, n is zero or an integer of 1 to 9, ^R2 is a hydrogen atom or a lower alkyl group when n is zero, and a hydrogen atom when n is an integer of 1 to 9; is the formula A group represented by the following may be formed, where R ³ represents a lower alkyl group, R ⁴ represents methyl or hydroxymethyl, R ⁵ represents hydrogen or methyl, R ⁶ represents carbamoyl, and R ⁷ represents a carboxyl group. ] and salts thereof. Furthermore, the present invention also provides an industrially advantageous method for producing the above compound () and its salt. Regarding the above general formula (), the lower alkyl groups as R, R ² and R ³ may be linear or branched and preferably have up to 6 carbon atoms, such as methyl, ethyl, n-propyl. , isopropyl, butyl, pentyl, hexyl, straight chain or branched at any position, and those having up to 4 carbon atoms are particularly preferred. In the general formula (), the alkyl or alkenyl having 3 to 41 carbon atoms represented by R ¹ may be linear or branched. In alkenyl,
The multiple bonds may be mutually free or conjugated. When it has a double bond, the main chain may be either trans or cis type via the double bond. Typical examples of alkyl or alkenyl groups having 3 to 41 carbon atoms represented by R ¹ are:

Examples of groups represented by the formula include acetyl, acryloyl, propionyl, methacryloyl, butyroyl, valeroyl, nonanoyl, trans-13-docosenoyl, palmitoyl, stearoyl, oleoyl, geranylacetyl, digeranylacetyl, and pharnesylacetyl. , geranylgeranyl acetyl, di(geranylgeranyl) acetyl, etc. Typical examples of the alkylene group having 2 to 9 carbon atoms represented by R ¹¹ include methylene, ethylene, propylene, butylene, and octylene. formula (In the formula, R ¹¹ has the same meaning as above.)

For example, 3-(2,3-dimethoxy-5
-Methyl-1,4-benzoquinone-6-yl)-
Propionyl, 6-(2,3-dimethoxy-5-methyl-1,4-benzoquinon-6-yl)-hexanoyl, 10-(2,3-dimethoxy-5-methyl-1,4-benzoquinon-6-yl) -decanoyl,6-(3,4-dimethoxy-5-methyl-
1,4-benzoquinon-6-yl)-4-methyl-4-hexenoyl, 4-(3,4-dimethoxy-5-methyl-1,4-benzoquinon-6-yl)-2-methyl-butyroyl, etc. be. Among the above-mentioned chain hydrocarbon groups, when the terminal is not substituted, a chain hydrocarbon group having 3 to 41 carbon atoms is mentioned, and the terminal is substituted with the above-mentioned 6-membered ring unsaturated hydrocarbon group. If the carbon number is 2 to 10
Examples include chain hydrocarbon groups. Also, as R ¹ the general formula

The compound () having a group represented by the formula: (in the formula, R ⁸ represents a hydrogen atom or a lower alkyl group) is also useful as a synthetic intermediate for high-molecular glucosamine derivatives, as described later. Among the compounds of the above general formula (), compounds in which R and R ³ are methyl groups are particularly preferred. Further, when R ⁴ is a hydrogen atom, R ⁵ is preferably a methyl group or a hydroxymethyl group, and when R ⁴ is a methyl group, R ⁵ is also preferably a methyl group. In the above compound (), the R ³ -substituted acetic acid residue bonded to the oxygen atom at the 3-position of the glucosamine residue is of the D type, and the R ⁴ , R ⁵ -substituted aminoacyl residues are R ⁴ , R When ⁵ is not both a hydrogen atom or a methyl group, the L type is preferable,
Preferably, the R ⁶ -substituted amino acid is of the D form. Compound () has acidity or neutrality depending on the type of substituent, and when it has acidity, it forms a salt with a base. Salts with bases include, for example, ammonium salts, salts with alkali metals or alkaline earth metals, such as with sodium, potassium, calcium or magnesium. Compounds () can be used as their physiologically acceptable salts. The compounds obtained according to the present invention have low toxicity,
Shows excellent immune function enhancing effect. In particular, the following experiments demonstrate that this compound enhances cellular immune responses, which are important for the protection of living organisms from infection, suppression of carcinogenesis, and treatment of cancer. The effect of this compound on enhancing cell-mediated immune responses in living organisms is, for example, its immune-enhancing effect on induction of delayed-type hypersensitivity to N-acetyltyrosine-3-azobenzene-4'-arsonic acid (ABA-N-Ac-Tyr) in guinea pigs. It is indicated by i.e. ABA-N-Ac-Tyr (50μg per animal)
A water-in-oil suspension prepared by dissolving this compound (10-100 μg per animal) in phosphate-buffered saline and mixing with Freund's incomplete adjuvant was applied to each footpad of Hartley guinea pigs.
Immunize by injecting 0.5ml each. Hair removal from the back for 2 weeks, ABA-bacteria-α-amylase (100μg)
is injected intradermally and the diameter of the intradermal reaction (erythema and induration) is measured 24 and 48 hours later. The size of this diameter is considered a measure of cell-mediated immunity. On the other hand, even if a water-in-oil suspension prepared by dissolving only the antigen ABA-N-Ac-Tyr in phosphate buffered saline and mixing it with Freund's incomplete adjuvant was administered, ABA-N- No delayed hypersensitivity to Ac-Tyr is observed. In this way, the compound of the present invention causes cell-mediated immune response (delayed type hypersensitivity).
It has enhanced immune-enhancing activity against. The cellular immunity-enhancing effect of this compound can also be recognized by its ability to significantly amplify the expression of cytotoxic reactions [expression of lymphocytes that exhibit specific damage to target cells (cancer cells)]. can. That is, mastocytoma P815-X2 (H-2 ^d ) was infected intraperitoneally in a C57BL/6J mouse (H-2b).
The cells are administered alone or together with the compound dissolved in phosphate-buffered saline for immunization. After immunization
On the 11th day, the spleen of the mouse was removed, and the amount of T-cells (killer T cells) that have cytotoxic activity against target cells produced in the spleen was measured using Brunner's method (Immunology Vol. 18, pp. 501-515). Measured by
When this compound is administered simultaneously with the above mastocytoma cells, the amount of killer T cells in the spleen is clearly increased. Cancer immunotherapy, which aims to eliminate non-self cancer cells by increasing the body's immune response ability, has been actively conducted in recent years, and killer T plays a major role in this.
The amount of killer T cells enhanced in this experiment can be considered as a measure of anti-cancer properties. Based on the above-mentioned effects, the compounds of the present invention can be used, for example, as infection-preventing agents, anticancer agents, etc., in the treatment of many diseases caused by a decrease in cellular immunity. On the one hand, this compound can be mixed with antigens to increase the immunogenicity of the antigens, so it is suitable for the production of diagnostic and therapeutic antisera as a mixture with various antigens, and on the other hand, it does not have to be added simultaneously with the antigens. They can also be used to stimulate immune responses that are already latent in the body. Therefore, in the case of chronic and acute infections, in the case of common immunodeficiencies, congenital or acquired, occurring during severe primary illnesses, such as in old age, and in patients with cancer, immunotherapy such as chemotherapy, radiotherapy, etc. It is particularly effective when undergoing treatment that causes functional decline, and is administered enterally to warm-blooded animals (humans; laboratory animals such as mice, guinea pigs, and rats; pet animals such as dogs and cats, etc.). For example, it can be administered orally or rectally, or parenterally. The dosage depends on the individual condition of the warm-blooded animal, species, age and mode of administration, but for example as an isotonic aqueous solution such as a salt-containing solution or a glucose solution for administration by injection; When administered by subcutaneous, intradermal, or intramuscular injection, the preferred dosage is about 1-500 μg/Kg/day of the compound (as anhydrous), especially about 5-500 μg/Kg/day.
It is 30μg/Kg/day. When administered by injection, the water-oil stable emulsion, preferably an oil, can also be administered as a vegetable or animal oil. For example, the vegetable or animal oil emulsion used is about 5-
It consists of 100 volumes of isotonic solution and about 1 volume of metabolizable vegetable or animal oil, as well as emulsion stabilizers. For administration via the oral route, it can be administered, for example, as a sugar-coated tablet or capsule, together with other excipients that can be used in the formulation, in which case the dosage ranges from about 40 to 4000 μg/Kg/day. be. In addition to the above-mentioned excellent biological activity, the compound of the present invention also has the advantage of being water-soluble, whereas conventional 6-O-acylglucosamine derivatives with anticancer activity are poorly water-soluble. . The compounds of the present invention are also useful as intermediates for the synthesis of other glucosamine derivatives. For example, as R ¹ the general formula

A compound () having a group represented by [Formula] (in the formula, R ⁸ represents a hydrogen atom or a lower alkyl group) [hereinafter referred to as compound (')
] can be used alone or with the general formula [In the formula, R ⁹ is a hydrogen atom or a lower alkyl group,
^R10 represents a carboxyl group that may be esterified, a hydroxyl group that may be etherified, or a pyrrolidino group that may be substituted. By doing so, Or above () and It is possible to obtain a polymer or a salt thereof having as a repeating unit. In this polymerization reaction, compound (') or compound (') and compound () can be used without a solvent or with a solvent such as halogenated hydrocarbons (carbon tetrachloride, chloroform, methylene chloride, etc.), ethers (tetrahydrofuran, dioxane, etc.). ), esters (methyl acetate, ethyl acetate, isoacyl acetate, etc.), organic acids (acetic acid, propionic acid, etc.), alcohols (methanol, ethanol, propanol, etc.), amides (dimethylformamide, etc.), sulfoxides (dimethyl sulfoxide, etc.) etc.) or water as a polymerization initiator, such as peroxides (benzoyl peroxide and its substituted products, tert-butyl hydroperoxaside, di-tert-butyl peroxide, potassium persulfate, ammonium persulfate, cumene hydroxyperoxide). Oxides, peroxides and transition metals or amines, or azo compounds such as azobisisobutyronitrile, azobiscyclohexanecarbonitrile, 2,2'-azobis(2-amidinopropane) hydrochloride, 2, 2′-
Azobis(2,4-dimethylvaleronitrile),
4,4'-azobis(4-cyano-1-pentanol) or 4,4'-azobis(4-cyanopentanoic acid), or organometallic compounds such as n-butyllithium, or inorganic salts such as aluminum chloride, dichloride, It is carried out using ditin, titanium tetrachloride, sodium naphthalene, boron trifluoride, diethyl ethylate, or acids such as phosphoric acid and perchloric acid. The reaction temperature usually ranges from room temperature (approx. 10-30℃) to approx.
The temperature is about 300°C, preferably about 30°-180°C. However, the reaction may be carried out at other temperatures if desired. The reaction pressure is usually normal pressure - about 700 kg/cm ² , and if desired, the reaction often proceeds more smoothly if the reaction is carried out under a zero nitrogen atmosphere. The polymer obtained in this way has an excellent immune function-enhancing effect like the compound (),
It is useful as an anti-infective agent, anti-cancer agent, etc., and can be used in the treatment of immunodeficiency diseases in the same manner as compound (). Compounds of the invention may, for example, have the general formula [In the formula, R, R ² , R ³ and n have the same meanings as above, and R ⁴ ′, R ⁵ ′, R ⁶ ′ and R ⁷ ′ are the above R ⁴ ,
A group represented by R ⁵ , R ⁶ , R ⁷ or a protected hydroxyl group or carboxyl group in these groups] and a compound represented by the general formula R ¹ —COOH () [wherein R ¹ has the same meaning as above] and by removing the protecting group as necessary. Regarding the above general formulas () and (), R ⁴ ′,
As the protecting group contained in the groups represented by R ⁵ ′, R ⁶ ′, and R ⁷ ′, any easily removable protecting group known per se in the fields of peptide chemistry and glycochemistry may be used, such as For carboxyl groups there may be mentioned inter alia tert-butyl, benzyl or benzhydryl groups, for hydroxyl groups there may be mentioned, for example, acyl groups such as lower alkanoyl groups (for example acetyl) or aroyl groups (for example benzoyl) or alkyl groups. (eg tert-butyl group) or tetrahydropyranyl group or benzoylcarbonyl group or lower alkanoylcarbonyl group, among others, benzyl which may be substituted with a halogen atom, a nitro group, a lower alkyl group or a lower alkoxy group, etc. or a benzylidene group or an optionally substituted lower alkylidene group. The condensation of the above compound () and compound () is
This can be carried out using condensation means known per se in the field of peptide chemistry. For example, it is preferable to activate the carboxyl group of compound () and react with compound (). Activated carboxyl groups include active esters and acid anhydrides. Examples of active esters include cyanomethyl ester, thioglycolic acid ester, p-nitrophenyl ester, 2,4,5-trichlorophenyl ester, pentachlorophenyl ester, pentafluorophenyl ester, and piparohydroxanoic acid ester. , N-hydroxyphthalimide ester, N-hydroxysuccinimide ester, N-hydroxy-5-norbornene-2,
3-dicarboximide ester, 8-hydroxyquinoline ester, 2-hydroxy-1,2-
Dihydro-1-carboethoxy-quinoline ester, 2-hydroxyphenyl ester, 2-hydroxy-4,5-dichlorophenyl ester, 2
-Hydroxypyridine ester, 2-pyridylthiol ester, 1-hydroxybenzotriazole ester unsubstituted or substituted with a halogenated methyl group or methoxy group, or N-ethyl-5-phenyl-isoxazolicam-3-sulfonate These are enol esters and the like that can be obtained once. Activated carboxyl groups are also optionally substituted with N-hydroxysuccinimide, N-hydroxy-5-norbornene-2,3-dicarboximide, unsubstituted or halogenated methyl or methoxy groups. Hydroxybenzotriazole, or 3-hydroxy-4
-Oxo-3,4-dihydro-benzo[d]-
By adding 1,2,3-triazine etc., N-N'-
It can also be obtained from dicyclohexylcarbodiimide. Preferred acid anhydrides include mixed acid anhydrides and acid amides such as imidazolide and isoxazolide. In the reaction of activated esters, organic bases such as triethylamine, N-methylmorpholine, N-ethylmorpholine or 1-hydroxybenzotriazole can also be present, if desired. The reaction temperature is usually about 0°C to about 80°C, preferably about 5° to 50°C. The reaction may be carried out at other temperatures if desired. The reaction generally proceeds in a solvent, and the solvent is
For example, halogenated hydrocarbons such as chloroform and carbon tetrachloride; ethers such as diethyl ether, tetrahydrofuran, and dioxane; esters such as ethyl acetate and isoamyl acetate; hydrocarbon groups such as benzene, toluene, and xylene; For example, nitriles such as acetonitrile, N-alkylamides such as N,N-dimethylformamide, N-methyl-2-pyrrolidone, dimethylacetamide, and other appropriate solvents such as dimethyl sulfoxide and hexamethylphosphoramide may be used. . The above compound () and compound () react stoichiometrically, but it goes without saying that these two components do not necessarily have to exist in an equivalent relationship in the reaction system; In contrast, about 1 to 5 moles, preferably about 1 to 2 moles of the compound () or its active derivative is used. The quantitative ratio of the two components depends on the combination of raw materials and other conditions.
It can be selected as appropriate to give good results. In addition, unreacted raw materials can be appropriately recovered and used as raw materials again. After the above reaction, the protecting group can be removed by a method known per se. It can be removed hydrolytically, for example by hydrogenation in the presence of a noble metal catalyst (eg platinum or palladium) or by acid hydrolysis. The compound () thus produced is isolated in a free state or as the above-mentioned salt using processing means known per se (e.g., extraction, dissolution, various chromatography, crystallization, reprecipitation, etc.). be able to. Note that the raw material compound has, for example, the general formula [In the formula, R ¹¹ represents a hydrogen atom or a protecting group,
R, R ³ , R ⁴ ′, R ⁵ ′, R ⁶ ′ and R ⁷ ′ have the same meanings as above] and the general formula [In the formula, n and R ² have the same meanings as above, and Y
represents a protecting group for an amino group] and can be easily produced by removing the protecting group. Regarding the above general formula (), as the protecting group for the amino group represented by Y, any easily removable protecting group known per se in the field of peptide chemistry may be used. For example, benzyloxycarbonyl (carbobenzoxy) group, p-methoxybenzylcarbonyl group, p-methylbenzylcarbonyl group, tert-butoxycarbonyl group, tert-amyloxycarbonyl group, p-biphenylisopropyloxycarbonyl group, O-nitro Phenylsulfenyl groups, trityl groups, etc. are conveniently used. The condensation reaction between compound () and compound () is
The condensation reaction between compound () and compound () described above can be carried out in the same manner as described above, and the protective group removal treatment after the reaction can also be carried out in the same manner as described above. Compound () can be prepared by, for example, "Green "Syutain and Winitz: Chemistry of the Amino Acids" 887-901
Each can be easily obtained by the method described in John Willey and Sons, Inc. (1961) or a method similar thereto. The present invention will be explained in more detail below using Examples and Reference Examples, but it goes without saying that these are not intended to limit the present invention. Example 1 (i) Benzyl 2-(benzyl 2-acetamido-2-deoxy-α-D-glucopyranoside-3-0-yl)-D-propionyl-L-alanyl-D-isoglutaminate (7.74 g,
11.5 mmol) was dissolved in N,N-dimethylformamide (30 ml), and carbobenzoxy-
β-alanine p-nitrophenyl ester (10.33g, 30mmol), 1-hydroxybenztriazole (3.11g, 23mmol), and N-ethylmorpholine (2.94ml, 23mmol) were added and stirred at room temperature for 2 days, and then the solvent was distilled off under reduced pressure. Dissolve the residue in ethyl acetate, add 5% sodium bicarbonate solution,
Wash sequentially with 1N hydrochloric acid and saturated saline, dry over anhydrous sodium sulfate, and then evaporate the solvent. The residue was purified by silica gel column chromatography.
Chloroform-acetone-methanol (10:
Purification with a mixed solvent of 3:2) yields benzyl, 2-
[Benzyl 2-acetamido-6-0-(carbobenzoxy-β-alanyl)-2-deoxy-α-D-glucopyranoside-3-0-yl)-D-propionyl-L-alanyl-D-
Obtain 77.12 g of isoglutamate powder. melting point
152-154゜〔α〕 ²³ _D +77.5゜ (c0.5, N,N-dimethylformamide) Rf = 0.48 (chloroform-methanol-acetic acid = 18:2:1, silica gel plate) (hereinafter Rf under these conditions Assuming Rf ¹ ) Elemental analysis value: C ₄₄ H ₅₅ O ₁₄ N ₅ Calculated value: C 60.19, H 6.31, N 7.98 Actual value: C 59.79, H 6.31, N 7.82 (ii) Benzyl 2-[benzyl 2-acetamido-6-0-(carbobenzoxy-β-alanyl)-2-deoxy-α-glucopyranoside-
3-0-yl]-D-propionyl-L-alanyl-D-isoglutaminate (4.39g,
Dissolve 5 mmol) in acetic acid (60 ml) and hydrogenate using palladium black as a catalyst at normal pressure and room temperature until the reaction stops. Separate the catalyst and wash it with a small amount of water.
The liquid was concentrated to dryness under reduced pressure, and the residue was applied to a column of Sephadex LH-20 and ethanol was added.
Developed and purified with a mixed solvent of 0.1N acetic acid (3:2) to obtain 2-(2-acetamido-6-0-β-alanyl-2-deoxy-α-D-glucose-3).
-0-yl)-D-propionyl-L-alanyl-D-isoglutamine (hereinafter referred to as 6-0-yl)-D-propionyl-L-alanyl-D-isoglutamine
β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine) is obtained quantitatively. Melting point 87°-91°C (decomposition), [α] ²³ _D +
33.2° (c 0.5, N, N-dimethyformamide) Rf = 0.32 (n-butanol-ethyl acetate-acetic acid-water = 1:1:1:1, silica gel plate) (Hereinafter, the Rf value under these conditions will be referred to as Rf ² . ) Elemental analysis value: C ₂₂ H ₃₇ O ₁₂ N ₅・12H ₂ O Calculated value: C 46.15, H 6.69, N 12.23 Actual value: C 46.46, H 6.99, N 11.95 (iii) Geranyl acetic acid (393 mg, 2 mmol) ) and p-nitrophenol (306 mg, 2.2 mmol) were dissolved in 4 ml of acetonitrile, and N,N'-dicyclohexylcarbodiimide (494 mg, 2.4 mmol) was dissolved under ice cooling.
and stir at 0°C for 1 hour and at room temperature for 15 hours. The precipitated crystals were removed and the solvent was distilled off under reduced pressure to obtain oily geranyl acetic acid p-nitrophenyl ester. (iv) 6-0-β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine (564
mg, 1 mmol) and geranyl acetic acid p-nitrophenyl ester (317 mg, 1.2 mmol) in
Dissolved in dimethylformamide (15 ml) and diluted with N-
Add ethylmorpholine (0.128ml) and bring to room temperature.
Stir for 17 hours. The solvent was distilled off under reduced pressure, and the residue was dissolved in a small amount of ethanol and acetic acid (0.1 ml).
was added, and then diethyl ether was added, and the precipitate formed was cooled and collected (700 mg). This was applied to a Sephadex LH-20 column, purified using 30% ethanol as a developing solvent, and 2-[2-acetamido-2-deoxy-
6-0-(geranyl acetyl-β-alanyl)
-D-glucose-3-0-yl)-D-propionyl-L-alanyl-D-isoglutamine
Get 415mg. [α] ²⁴ _D +29.9° (after 3 minutes) → 28.2° (after 25 hours) [c 0.77, water]. Rf ² =0.65, Rf=
0.47 (ethyl acetate-pyridine-acetic acid-water = 30:
10:3:5, silica gel plate) (hereinafter, the Rf value under this condition will be referred to as Rf ³ ). Elemental analysis value: C ₃₄ H ₅₅ O ₁₃ N ₅ H ₂ O Calculated value: C 53.74, H 7.56, N 9.21 Actual value: C 53.31, H 7.43, N 8.96 (v) 2-[2-acetamide-2- Deoxy-6
-0-(geranylacetyl-β-alanyl)-
D-glucose-3-0-yl]-D-propionyl-L-alanyl-D-isoglutamine
Dissolve 150mg, 0.2mmol) in 2ml of water. Add 0.5N caustic soda dropwise to the ice-cooled container while stirring to adjust the pH to 7, and repeat freeze-drying twice.
-acetamido-2-deoxy-6-0-(geranylacetyl-β-alanyl)-D-glucose-3-0-yl]-D-propionyl-L
-Alanyl-D-isoglutamine Na salt 162mg
get. Elemental analysis value: C ₃₄ H ₅₄ O ₁₃ N ₅ Na・2H ₂ O Calculated value: C 51.06, H 7.31, N 8.76 Actual value: C 50.81, H 7.66, N 8.21 Example 2 (i) Digeranyl acetic acid (300 mg , 0.9 mmol) and p-
Nitrophenol (139 mg, 1 mmo) was dissolved in a mixed solvent of acetonitrile-ethyl acetate (1:1) (10 ml), and N,N'-dicyclohexylcarbodiimide (227 mg, 1.1 mmol) was added under ice cooling, and the mixture was heated at 0°C for 3 hours. Stir at room temperature for 15 hours.
The precipitated crystals were removed and the solvent was distilled off under reduced pressure to obtain digeranyl acetic acid p-nitrophenyl ester. (ii) The above digeranyl acetic acid p-nitrophenyl ester and 6-0-β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine (423 mg, 0.75 mmol) were combined with N,N-dimethylformamide ( 10 ml), add N-ethylmorpholine (0.1 ml), and react at room temperature for 2 days. The solvent is distilled off under reduced pressure, the residue is dissolved in ethanol, and a small amount of insoluble material is filtered off. Add acetic acid (0.1 ml) to the filtrate, then add ethyl ether, and collect the resulting precipitate by filtration (500 mg). This was applied to a silica gel column, developed with a mixed solvent of ethyl acetate, pyridine, acetic acid, and water (30:10:3:5), and the eluted fraction of the main product was collected, and the solvent was removed under reduced pressure to approximately 5 ml. Concentrate, add 100 ml of water, lyophilize, and cephadex the residue (130 mg).
Apply to LH-20 column and purify with 60% ethanol to produce 2-[2-acetamide-6-0-
(Digeranylacetyl-β-alanyl)-2-
Deoxy-D-glucose-3-0-yl]-
91 mg of D-propionyl-L-alanyl-D-isoglutamine is obtained. [α] ²⁴ _D +25.0° (after 3 minutes)
→+24.7° (24 hours later) [c 0.32, water], Rf ² =
0.80, Rf ³ = 0.59 Elemental analysis value: C ₄₅ H ₇₁ O ₁₅ N ₅・H ₂ O Calculated value: C 59.48, H 7.82, N 7.45 Actual value: C 57.07, H 7.81, N 7.67 Example 3 (i ) Falnesyl acetic acid (52.9 mg, 0.2 mmol) and N
-Hydroxy-5-norbornene-2,3-dicarboximide (hereinafter referred to as HONB)
(39.4 mg, 0.22 mmol) in acetonitrile (2
ml), add N,N'-dicyclohexylcarbosiimide (45.4 mg, 0.22 mmol) under ice-cooling, and stir at 0°C for 2 hours and at room temperature for 13 hours. The precipitated crystals are removed, and the solvent is distilled off under reduced pressure to obtain oily FARNSYL acetic acid HONB ester. (ii) The above falnesyl acetic acid HONB ester and 6-0-β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine (112.7
mg, 0.2 mmol) in N,N-dimethylformamide (4 ml), add N-ethylmorpholine (25.6 μ), and stir at room temperature for 17 hours. The solvent was distilled off under reduced pressure, and the residue was applied to a silica gel column and ethyl acetate-pyridine-acetic acid-
Develop with a mixed solvent of water (30:10:3:5), collect the eluted fraction of the main product, and evaporate the solvent under reduced pressure. The residue was applied to a Sephadex LH-20 column, and ethanol-0.1N acetic acid (3:2) was used.
Elute with a mixed solvent of 2-[2-acetamido-2-deoxy-6-0-(phalnesylacetyl-β-alanyl)-D-glucose-3-
80.2 mg of 0-yl)-D-propionyl-L-alanyl-D-isoglutamine is obtained. [α] ²³ _D +
38.4° (c 0.5, N, N-dimethylformamide). Rf ² = 0.72, Rf ³ = 0.53 Elemental analysis value: C ₃₉ H ₆₃ O ₁₃ N ₅・H ₂ O Calculated value: C 56.57, H 7.91, N 8.45 Actual value: C 56.24, H 7.96, N 8.43 Example 4 Geranylgeranyl acetic acid 83.1mg (0.25mmol)
HONB (50.4 mg, 0.28 mmol) and N,N'-dicyclohexylcarbosiimide (57.7 mg, 0.28 mmol)
and are reacted in acetonitrile (5 ml) in the same manner as in Example 3 (i). The generated active ester and 6
-0-β-alanyl-N-acetylmuramyl-L
-Alanyl-D-isoglutamine (141mg,
0.25 mmol) in N,N-dimethylformamide (4 ml), N-ethylmorpholine (36μ,
0.28 mmol) and react at room temperature for 60 hours in the dark. This was purified in the same manner as in Example 3 (ii), and 2-[2-acetamido-2-deoxy-
6-0-(geranylgeranyl acetyl-β-alanyl)-D-glucose-3-0-yl]-D-
74 mg of propionyl-L-alanyl-D-isoglutamine is obtained. [α] ²⁷ _D +25.8゜ (c 0.3, 70% ethanol) Rf ³ = 0.70 Elemental analysis value: C ₄₄ H ₇₁ O ₁₃ N ₅・2H ₂ O Calculated value: C 57.81, H 8.27, N 7.66 Actual measurement Values: C 57.59, H 7.95, N 7.62 Example 5 Di-(geranylgeranyl)acetic acid (91 mg,
0.15mmol) and HONB (32.4mg, 0.18mmol) and N,
N′-dicyclohexylcarbodiimide (37.1mg,
(i) of Example 3 in a mixed solvent (4 ml) of acetonitrile-ethyl acetate (1:1).
Treat the reaction in the same manner as above. The obtained active ester and 6-0-β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine (85 mg,
0.15 mmol) in N,N-dimethylformamide (2.5 ml) with the addition of N-ethylmorpholine (0.02 ml) and reacted at room temperature for 48 hours. Example 3
Purify in the same manner as in (ii) to obtain 2-{2-acetamido-2-deoxy-6-0-[geranylgeranyl)acetyl-β-alanyl]-D-glucose-3-0-yl}-D-propionyl. -49 mg of L-alanyl-D-isoglutamine is obtained. [α] ²⁷ _D +
20.0° (c 0.3, 70% ethanol). Rf ³ = 0.86 Elemental analysis value: C ₆₄ H ₁₀₃ O ₁₃ N ₅・2H ₂ O Calculated value: C 64.78, H 9.09, N 5.90 Actual value: C 64.58, H 9.08, N 6.00 Example 6 Stearic acid p- Nitrophenyl ester 486
mg (1.2 mmol) and 6-0-β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine (563.5 mg, 1 mmol) were dissolved in N,N-dimethylformamide (10 ml) and N- Add ethylmorpholine (0.18 ml) and stir at room temperature for 17 hours. The solvent was distilled off under reduced pressure. Ethyl acetate was added to the residue to slightly dissolve it, then ethyl ether was added and cooled, the precipitate was collected and applied to a silica gel column, and ethyl acetate-pyridine-acetic acid-water (100:30: Developed with a mixed solvent of 9:15, V/V) to obtain 550 mg of crude product. 70mg of this
was purified using a mixed solvent of ethanol and 0.1N acetic acid (3:2) on a Cephadex LH-20 column to obtain 2-[2-acetamido-2-deoxy-6-
0-(stearoyl-β-alanyl)-D-glucose-3-0-yl]-D-propionyl-L
- Obtain 60 mg of alanyl-D-isoglutamine.
[α] ²⁷ _D +22,40 (after 3 minutes) → 28.1° (after 25 hours) [
c.
0.5, 70% ethanol]. Rf ³ = 0.67 Elemental analysis value: C ₄₀ H ₇₁ O ₁₃ N ₅・2H ₂ O Calculated value: C 55.47, H 8.73, N 8.09 Actual value: C 55.71, H 8.88, N 8.12 Example 7 Trans-13- Docosenoic acid (101.6mg,
0.3 mmol) and HONB (59.1 mg, 0.33 mmol) and N,
N′-dicyclohexylcarbodiimide (68.1mg,
0.33 mmol) in acetonitrile (4 ml) in the same manner as in Example 3 (i). The obtained active ester and 6-0-β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine (169.1 mg, 0.3 mmol) were dissolved in N,N-dimethylformamide (3 ml). -React at room temperature for 48 hours in the presence of ethylmoline (38.4μ, 0.3mmol). This was purified in the same manner as in Example 3 (ii), and 2-[2-acetamido-2-deoxy-6
-0-(trans-13-docosenoyl-β-alanyl)-D-glucose-3-0-yl]-D-
123.8 mg of propionyl-L-alanyl-D-isoglutamine is obtained. [α] ²³ _D +32.2 (c 1.0 ethanol), Rf ² =0.77, Rf ³ =0.61. Elemental analysis value: C ₄₄ H ₇₇ O ₁₃ N ₅ 3/2H ₂ O Calculated value: C 58.00, H 8.85, N 7.67 Actual value: C 58.11, H 8.83, N 7.65 Example 8 3-(2,3- dimethoxy-5-methyl-1,
4-Benzoquinone-6-yl)-propionic acid (76.3 mg, 0.3 mmol) and HONB (59.1 mg,
Dissolve 0.3 mmol) in acetonitrile (3 ml),
N,N'-dicyclohexylcarbodiimide (68.1 mg, 0.3 mmol) was added under ice cooling, and the mixture was heated at 0°C for 1 hour.
React for 4 hours at room temperature. The precipitated crystals were removed and the solvent was distilled off under reduced pressure. The obtained active ester and 6-0-β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine (169.1 mg,
0.3 mmol) in N,N-dimethylformamide (3
ml) and N-ethylmorpholine (38.4μ,
0.3 mmol) and stir at room temperature for 17 hours.
This was purified in the same manner as in Example 3 (ii), and 2-
{2-acetamido-2-deoxy-6-0-
[3-(2,3-dimethoxy-5-methyl-1,
4-benzoquinone-6-yl)-propionyl-
134 mg of β-alanyl]-D-glucose-3-0-yl}-D-propionyl-L-alanyl-D-isoglutamine is obtained. All post-reaction treatments are performed in the dark. [α] ²³ _D +31.2゜(c 0.5, N, N-
dimethylformamide). Rf ² = 0.63, Rf ³ = 0.34 Elemental analysis value: C ₃₄ H ₄₉ O ₁₇ N ₅・3H ₂ O Calculated value: C 47.82, H 6.49, N 8.20 Actual value: C 47.66, H 6.11, N 8.23 Example 9 10-(2,3-dimethoxy-5-methyl-1,
4-Benzoquinone-6-yl)-decanoic acid (70.5
mg, 0.2 mmol) and p-nitrophenol (30.6
mg, 0.22 mmol) and N,N'-dicyclohexylcarbodiimide (45.4 mg, 0.22 mmol) in acetonitrile (4 ml) as in (iii) of Example 1. The obtained active ester and 6-0-
β-alanyl-N-acetylmuramyl-L-alanyl-D-isoglutamine (112.7mg, 0.2mmol)
and N,N-dimethylformamide (4 ml)
Medium, N-ethylmorpholine (25.6μ, 0.2mmol)
React for 24 hours at room temperature in the presence of The solvent was distilled off under reduced pressure, and the residue was applied to a column of Sephadex, LH-20, and mixed with ethanol-0.1N acetic acid (3:2).
Elute with a mixed solvent of 2-{2-acetamide-
2-deoxy-6-0-[10-(2,3-dimethoxy-5-methyl-1,4-benzoquinone-6-
0-yl)-decanoyl-β-alanyl]-D-
96 mg of glucose-3-0-yl}-D-propionyl-L-alanyl-D-isoglutamine is obtained. All post-reaction treatments are performed in the dark. [α] ²³ _D +
29.0° (c 0.5, ethanol). Rf ² = 0.72, Rf ³ =
0.42 Elemental analysis value: C ₄₁ H ₆₃ O ₁₇ N ₅ · H ₂ O Calculated value: C 54.29, H 7.22, N 7.72 Actual value: C 54.13, H 7.21, N 7.50 Example 10 Retinoic acid [3,7- Dimethyl-9-(2,
6,6-trimethyl-1-cyclohexene-1-
yl)-2,4,6,8-nonatetraenoic acid]
(150.2 mg, 0.5 mmol) and HONB (104.4 mg,
0.58 mmol) in a mixed solvent (4 ml) of acetonitrile-ethyl acetate (1:1), add N,N'-dicyclohexylcarbodiimide (119.5 mg, 0.58 mmol) under ice-cooling, and stir at 4°C for 17 hours. The precipitated crystals were removed and the solvent was distilled off under reduced pressure. The obtained active ester and 6-0-β-alanyl-N-acetylmuramyl-L-alanyl-
D-isoglutamine (282 mg, 0.5 mmol) and N,
Dissolve in 6 ml of N-dimethylformamide, add N-ethylmorpholine (0.1 ml), and react at 4°C for 48 hours. This was purified in the same manner as in Example 6,
2-[2-acetamido-2-deoxy-6-0
115 mg of -(retinoyl-β-alanyl)-D-glucose-3-0-yl]-D-propionyl-L-alanyl-D-isoglutamine is obtained. All post-reaction treatments are performed in a cool, dark place. [α] ²⁷ _D +21.6° (after 3 minutes) → +23.9° (after 25 hours) [c 0.5, water].
Rf ³ = 0.41 Elemental analysis value: C ₄₂ H ₆₃ O ₁₃ N ₅・5H ₂ O Calculated value: C 53.85, H 7.86, N 7.48 Actual value: C 53.47, H 7.19, N 7.43 Example 11 (i) N -t-butyloxycarbonyl-0-benzyl-L-serine (4.43g, 15mmol) and D-isoglutamine benzyl ester (3.54g,
15 mmol) in acetonitrile (100 ml), and add N,N'-dicyclohexylcarbodiimide (3.50 g, 17 mmol) under ice cooling. After stirring for 3 hours under ice cooling and 15 hours at room temperature, the precipitate was removed and the solvent was distilled off. The residue was dissolved in ethyl acetate (100ml)
Dissolved in water, washed sequentially with 1N hydrochloric acid, 5% aqueous sodium bicarbonate, and water, dried over anhydrous sodium sulfate, and evaporated the solvent. The residue was crystallized from ethyl acetate-petroleum ether to obtain 3.40 g of Nt-butyloxycarbonyl-0-benzyl-L-seryl-D-isoglutamine benzyl ester. Melting point 65-66℃.
[α] ²³ _D +57.7° (c 0.5, ethanol). Rf ¹ =
0.73. Elemental analysis value C ₂₇ H ₃₅ O ₇ N ₃ Calculated value: C 63.14, H 6.87, N 8.18 Actual value: C 62.89, H 6.93, N 8.19 (ii) Benzyl, 2-acetamide-4,6-0-
Benzylidene-3-0-(D-1-carboxyethyl)-2-deoxy-α-D-glucopyranoside 7.07 (15 mmol) and HONB (3.22 g, 18 propionyl) were dissolved in 110 ml of tetrahydrofuran and cooled with N,N' under ice cooling. - Add dicyclohexylcarbodiimide (3.71g, 18mmol) and incubate at 0℃ for 30 minutes.
Incubate the reaction for 16 hours at room temperature. The precipitated crystals were removed and the solvent was distilled off under reduced pressure. The residue was recrystallized from a mixed solution of ethyl acetate and ether to give benzyl 2-acetamido-4,6-0-benzylidene-3-0-(D-1-carboxyethyl)-2-deoxy-α-D-glucopyranoside. Obtain 7.42 g of HONB active ester. Melting point 122
-124℃ (iii) Dissolve N-t-butyloxycarbonyl-0-benzyl-L-seryl-D-isoglutamine benzyl ester (5.0 g, 10 mmol) in trifluoroacetic acid (25 ml) and leave at room temperature for 20 minutes. . Distill trifluoroacetic acid. A mixture of petroleum ether and diethyl ether (1:1) was added to the residue to remove the precipitate. Dissolve this in N,N-dimethylformamide (20 ml), and add benzyl 2- to this solution under ice cooling with triethylamine (1.60 ml) and acetonitrile (50 ml).
Acetamide-4,6-0-benzylidene-3
-0-(D-1-carboxyethyl)-2-deoxy-α-D-glucopyranoside HONB
Add active ester (6.35g, 10mmol). After standing at room temperature for 60 hours, diethyl ether (100 ml) was added to collect the precipitated crystals. This was recrystallized from N,N-dimethylformamide-acetonitrile to give benzyl 2-(benzyl-2
-acetamide-4,6-0-benzylidene-
2-deoxy-α-D-glucopyranoside-3
-0-yl)-D-propionyl-0-benzyl-L-seryl-D-isoglutaminate
Get 7.60g. Melting point 259℃ (decomposition). [α] ²³ _D +
81.1° (c 0.5, N, N-dimethylformamide). Rf ¹ = 0.69 Elemental analysis value C ₄₇ H ₅₄ O ₁₂ N ₄ Calculated value: C 65.11, H 6.28, N 6.46 Actual value: C 65.23, H 6.21, N 6.40 (iv) Benzyl 2-(Benzyl 2-acetamide- 4,6-0-benzylidene-2-deoxy-α-D-glucopyranoside-3-0-yl)
-D-propionyl-0-benzyl-L-seryl-D-isoglutaminate (7.50g,
8.65 mmol) was dissolved in 75% acetic acid by heating at 100°C, and after further heating at 100°C for 1 hour, the solvent was distilled off. The crystalline residue was recrystallized from ethanol-diethyl ether to give benzyl 2-(benzyl 2-
4.48 g of acetamido-2-deoxy-α-D-glucopyranoside-3-0-yl)-D-propionyl-0-benzyl-L-seryl-D-isoglutaminate is obtained. Melting point 188-191℃.
[α] ²³ _D +48.8° (c 0.5, N, N-dimethylformamide). ^Rf1 =0.31. Elemental analysis value: C ₄₀ H ₅₀ O ₁₃ N ₄・1/2H ₂ O Calculated value: C 60.98, H 6.53, N 7.11 Actual value: C 61.07, H 6.38, N 7.00 (v) Benzyl: C 2-( Benzyl 2-acetamido-2-deoxy-α-D-glucopyranoside-3-0-yl)-D-propionyl-0
-Benzyl-L-seryl-D-isoglutamate (3.12mg, 4mmol) and carbobenzoxy-
L-leucine p-nitrophenyl ester (2.32 g, 6 mmol), N-ethylmorpholine (1.54 ml, 12 mmol) and 1-hydroxybenztriazole (1.62 g, 12 mmol) were dissolved in N,N-
Dissolve in dimethylformamide (10 ml) and react at room temperature for 90 hours. Water (100 ml) was added to the reaction solution, extracted with ethyl acetate (150 ml), and the ethyl acetate layer was washed successively with 1N ammonium water, 1N hydrochloric acid, and water, dried over anhydrous sodium sulfate, and the solvent was distilled off. The residue was purified by applying it to a silica gel column and developing with a mixed solvent of chloroform-methanol (19:1) to obtain benzyl 2-[benzyl
2-acetamido-6-0-(carbobenzoxy-L-leucyl)-2-deoxy-α-D-
2.55 g of glucopyranoside-3-0-yl]-D-propionyl-0-benzyl-L-seryl-D-isoglutaminate are obtained as crystals. Melting point 177-178℃. [α] ²³ _D +58.7゜(c 0.5, N, N
-dimethylformamide). ^Rf1 =0.70. Elemental analysis value C ₅₄ H ₆₇ O ₁₅ E ₅ Calculated value: C 63.20, H 6.58, N 6.83 Actual value: C 63.01, H 6.49, N 6.71 (vi) Benzyl 2 [benzyl 2-acetamide-6-0-( Carbobenzoxy-L-leucyl)-2-deoxy-α-glucopyranoside
3-0-yl]-D-propionyl-0-benzyl-L-ceryl-D-isoglutaminate (2.00 g, 1.95 mmol) is hydrogenated in acetic acid (30 ml) over palladium black as a catalyst for 12 hours. After removing the catalyst, the solvent was distilled off. The residue was reprecipitated from ethanol-diethyl ether to give 2(2-acetamido-2-deoxy-6-0-L-leucyl-D-glucose-3-0-yl)-D-
1.22 g of propionyl-L-seryl-D-isoglutamine is obtained. Melting point: 154℃ (decomposition). [α] ²³ _D +
21.0° (5 after 3 minutes → +28.8° (after 25 hours) (c 0.5,
water). Rf ² = 0.55 Elemental analysis value C ₂₅ H ₄₃ O ₁₃ N ₅・H ₂ O Calculated value: C 46.93, H 7.09, N 10.94 Actual value: C 47.00, H 7.19, N 10.52 (vii) 10-(2, 3-dimethoxy-5-methyl-
1,4-Benzoquinone-6-yl)-decanoic acid (33.8 mg, 0.1 mmol), p-nitrophenol (16.7 mg, 0.12 mmol) and N,N-dicyclohexylcarbodiimide (24.7 mg, 0.12 mmol) were dissolved in acetonitrile ( 2 ml) was reacted and treated in the same manner as in Example 8. The obtained active ester and 2-(2-acetamido-2-deoxy-6-
0-L-leucyl-D-glucose-3-0-
Il)-D-propionyl-L-seryl-D-
Isoglutamine (62.1mg, 0.1mmol) and N,
The mixture was reacted in N-dimethylformamide (2 ml) in the presence of N-ethylmorpholine (20μ) at room temperature for 24 hours, and the solvent was distilled off. The residue was purified in the same manner as in Example 3 (ii) to obtain 2-[2-acetamide-2
-deoxy-6-0-[10-(2,3-dimethoxy-5-methyl-1,4-benzoquinone-6
-Il) -Decanoyl-L-Leucil] -D-
48.4 mg of glucose-3-0-yl]-D-propionyl-L-seryl-D-isoglutamine is obtained. [α] ²³ _D +17.1° (after 3 minutes) → +16.7° (after 25 hours) [c 0.5, 70% ethanol]. ^Rf3 =0.50. Elemental analysis value C ₄₄ H ₆₉ O ₁₅ N ₅・2H ₂ O Calculated value: C 53.27, H 7.42, N 7.06 Actual value: C 53.40, H 7.09, N 7.05 Example 12 (i) Carbobenzoxy-L- Proline p-nitrophenyl ester (1.48g, 4mmol) and benzyl 2-(benzyl 2-acetamide-2)
-Deoxy-α-D-glucopyranoside-3-
0-yl)-D-propionyl-L-alanyl-D-isoglutaminate (1.34g, 2mmol)
and N,N-dimethylformamide (10
ml). Further refine in the same way,
Benzyl 2-[benzyl 2-acetamide-6-0-(carbobenzoxy-L-prolyl)-2-deoxy-α-glucopyranoside-
732 mg of 3-0-yl]-D-propionyl-L-alanyl-D-isoglutaminate are obtained.
Melting point 110-112℃ [α] ²³ _D +61.1゜(c 0.5, N,
N dimethylformamide). ^Rf1 =0.57. Elemental analysis value C ₄₆ H ₅₇ O ₁₄ N ₅ Calculated value: C 61.11, H 6.36, N 7.75 Actual value: C 61.15, H 6.45, N 7.53 (ii) Benzyl 2-[benzyl 2-acetamide-6-0- (Carbobenzoxy-L-prolyl)-2-deoxy-α-D-glucopyranoside-3-0-yl]-D-propionyl-L-alanyl-D-isoglutaminate (482 mg,
0.53 mmol) was hydrogenated in the same manner as in Example 1 (ii) in the presence of palladium black in acetic acid. Thereafter, the same post-purification treatment was carried out to obtain 2-(2-acetamide-
2-deoxy-6-0-L-prolyl-D-glucose-3-0-yl)-D-propionyl-L-alanyl-D-isoglutamine is quantitatively obtained. Melting point: 159℃ (decomposition). [α] ²³ _D +37.6゜(c
0.5,N,N-dimethylformamide). Rf ²
=0.29. Elemental analysis value C ₂₄ H ₃₉ O ₁₂ N ₅・1.5H ₂ O Calculated value: C 46.75, H 6.87, N 11.35 Actual value: C 47.06, H 7.00, N 10.77 (iii) 3-(2,3-dimethoxy -5-methyl-
1,4-Benzoquinone-6-yl)-propionic acid (38.1 mg, 0.15 mmol), p-nitrophenol (22.9 mg, 0.165 mmol), and N,N'-dicyclohexylcarbodiimide (34 mg,
0.165 mmol) in acetonitrile (2 ml),
The reaction and treatment are carried out in the same manner as in Example 8. The obtained active ester and 2-(2-acetamido-2-deoxy-6-0-L-prolyl-D-
glucose-3-0-yl)-D-propionyl-L-alanyl-D-isoglutamine (88.5
mg, 0.15 mmol) in N,N-dimethylformamide (2 ml) in the presence of N-ethylmorpholine (30μ) at room temperature for 24 hours, and the solvent was distilled off. The residue was purified in the same manner as in Example 3 (ii) to obtain 2-{2-acetamido-2-deoxy-6-0-[3-(2,3-dimethoxy-5-
Methyl-1,4-benzoquinon-6-yl)-
Propionyl-L-prolyl]-D-glucose-3-0-yl}-D-propionyl-L-
63 mg of alanyl-D-isoglutamine is obtained.
[α] ²³ _D +10.4° (after 3 minutes) → +10.4° [c 0.5, 70
%ethanol〕. ^Rf3 =0.41. Elemental analysis value C ₃₆ H ₅₁ O ₁₇ N ₁₅・3H ₂ O Calculated value: C 49.13, H 6.53, N 7.96 Actual value: C 49.22, H 5.98, N 8.05 Example 13 (i) 10-(2,3 -dimethoxy-5-methyl-
1,4-Benzoquinone-6-yl)-decanoic acid (96 mg, 0.27 mmol) and p-nitrophenol (41.7 mg, 0.3 mmol) were dissolved in acetonitrile (2
ml), add N,N'-dicyclohexylcarbodiimide (61.9 mg, 3 mmol) under ice-cooling, and stir at 4°C for 16 hours. After removing the precipitate, the solvent was distilled off under reduced pressure. The residue was recrystallized from a mixed solvent of ethyl acetate and petroleum ether.
-(2,3-dimethoxy-5-methyl-1,4
-Benzoquinone-6-yl)-decanoic acid p-
Obtain 120 mg of nitrophenyl ester. melting point 52
-54℃ Elemental analysis value C ₂₅ H ₃₁ O ₈ N Calculated value: C 63,14, H 6.60, N 2.96 Actual value: C 63.49, H 6.84, N 3.39 (ii) Benzyl 2-(benzyl 2-acetamide- 2-deoxy-α-D-glucopyranoside-3-0-yl)-D-propionyl-L-alanyl-D-isoglutaminate (3.36g,
5mmol) in N,N-dimethylformamide (30ml). In this, carbobenzoxy-ε
-Amino-n-caproic acid p-nitrophenyl ester (3.86, 10 mmol) and 1-hydroxybenztriazole (2.70 g, 20 mmol)
Melt it, add 2.6 ml of triethylamine under ice cooling, and stir at room temperature for 16 hours. Distill the liquid under reduced pressure except for the precipitate. The residue was dissolved in ethyl acetate, washed sequentially with 5% aqueous sodium bicarbonate, 0.5N hydrochloric acid, and water, dried over anhydrous sodium sulfate, and the solvent was distilled off.
The residue was purified by applying it to a silica gel column and developing with chloroform-acetone-methanol (10:3:1), and then reprecipitated from ethyl acetate-diethyl ether to give benzyl 2-[benzyl 2-acetamide-6- 0-(carbobenzoxy-ε-amino-n-caproyl)-2
-Deoxy-α-D-glucopyranoside-3-
1.27 g of 0-yl]-D-propionyl-L-alanyl-D-isoglutaminate are obtained. melting point
161-162℃. [α] ²³ _D +74.8゜(c 0.5, N, N-
dimethylformamide]. ^Rf1 =0.54. Elemental analysis value C ₄₇ H ₆₁ O ₁₄ N ₅ Calculated value: C 61,36, H 6.68, N 7.61 Actual value: C 61.49, H 6.69, N 7,37 (iii) Benzyl 2 [benzyl 2-acetamide-6 -0- [carbobenzoxy-ε-amino-
n-caproyl)-2-deoxy-α-D-glucopyranoside-3-0-yl]-D-propionyl-L-alanyl-D-isoglutaminate (374 mg, 0.4 mmol) was added to (ii) of Example 1. In the same manner, it was hydrogenated in the presence of palladium black in acetic acid and purified to produce 2-(2-acetamide-6-
202 mg of 0-ε-amino-n-caproyl-2-deoxy-D-glucose-3-0-yl)-D-propionyl-L-alanyl-D-isoglutamine is obtained. Melting point 65-97℃. [α] ²³ _D +24.0° (c 0.5, N-dimethylformamide). ^Rf2 =
0.40. Elemental analysis value C ₂₅ H ₄₃ O ₁₂ N ₅・3/2H ₂ O Calculated value: C 47.46, H 7.33, N 11.07 Actual value: C 47.50, H 7.34, N 10.57 (iv) 2-(2-acetamide- 6-0-ε-amino-n-caproyl-2deoxy-D-glucose-3-0-yl)-D-propionyl-L
-Alanyl-D-isoglutamine (75.7mg,
Dissolve 125μmol) in N,N-dimethylformamide (1ml) and add N-ethylmorpholine (16μ) under ice cooling. Add 10-(2,
3-Dimethoxy-5-methyl-1,4-benzoquinon-6-yl)-decanoic acid p-nitrophenyl ester (29.2 mg, 125 μmol) was added, and the mixture was allowed to react at room temperature for 16 hours. This was purified in the same manner as in Example 3 (ii), and 2-{2-acetamide-
2-deoxy-6-0-[10-(2,3-dimethoxy-5-methyl-1,4-benzoquinone-
6-yl)-decanoyl-ε-amino-n-caproyl]-D-glucose-3-0-yl}
-94 mg of D-propionyl-L-alanyl-D-isoglutamine is obtained. [α] ²³ _D +28.6 (c 0.5,
Ethanol) Rf ² = 0.76, Rf ³ = 0.52 Elemental analysis value C ₄₄ H ₆₈ O ₁₇ N ₅ · H ₂ O Calculated value: C 55.15, H 7.47, N 7.31 Actual value: C 54.97, H 7.24, N 7.25 Implemented Example 14 (i) 3-(2,3-dimethoxy-5-methyl-
Dissolve 1,4-benzoquinone-6-yl)-propionic acid (38.1 mg, 0.15 mmol) and p-nitrophenol (23.0 mg, 0.165 mmol) in acetonitrile (2 ml), and add N,N'-dicyclohexylcarbodiimide under ice cooling. (34.0mg,
0.165 mmol) and stir at 4°C for 16 hours. After removing the precipitate, the solvent was distilled off under reduced pressure and 3-(2,3-dimethoxy-5-methyl-
1,4-Benzoquinone-6-yl)-propionic acid p-nitrophenyl ester is obtained. (ii) Benzyl 2-(benzyl 2-acetamido-2-deoxy-α-D-glucopyranoside-3-0-yl)-D-propionyl-L-alanyl-D-isoglutaminate (1.35 g,
Dissolve 2 mmol) in N,N-dimethylformamide (15 ml) and add carbobenzoxy-
Glycine p-nitrophenyl ester (1.32 g, 4 mmol), 1-hydroxybenztriazole (1.08, 8 mmol) and N-ethylmorpholine (1.02 ml, 8 mmol) were added at room temperature.
After stirring for several days, the solvent was distilled off under reduced pressure. Dissolve the residue in ethyl acetate, add 5% sodium bicarbonate solution,
Wash sequentially with 0.2N hydrochloric acid and water, dry over anhydrous sodium sulfate, and evaporate the solvent. The residue was purified by precipitating it as a gel from a mixed solvent of ethyl acetate-ether-petroleum ether to give benzyl 2-[benzyl 2-acetamide-6-0-(carbobenzoxy-glycyl)-2-deoxy-α]. ―
D-glucopyranoside-3-0-yl]-D-
1.13 g of propionyl-L-alanyl-D-isoglutamate are obtained. Melting point 192-193℃. [α]
²³ _D +80.2° (c 0.5, N, N-dimethylacetamide). Rf ¹ = 0.47 Elemental analysis value: C ₄₃ H ₅₃ O ₁₄ N ₅ Calculated value: C 59, 78, H 6.18, N 8.11 Actual value: C 59.65, H 6.01, N 7.96 (iii) Benzyl 2 - [Benzyl 2 -acetamido-6-0-(carbobenzoxy-glycyl)
-2-deoxy-α-D-glucopyranoside-
3-0-yl]-D-propionyl-L-alanyl-D-isoglutamate (864 mg,
1 mmol) was hydrogenated in acetic acid using palladium black as a catalyst in the same manner as in (ii) of Example 1, and then purified to give 2-(2-acetamido-2-deoxy-6-0-glycyl-D-glucose). -3-
464 mg of 0-yl)-D-propionyl-L-alanyl-D-isoglutamine is obtained. Melting point 104-
107℃. [α] ²³ _D +48.0° (c 0.5, N, N-dimethylformamide). Rf ³ = 0.32 Elemental analysis value: C ₂₁ H ₃₅ O ₁₂ N ₅・1/2H ₂ O Calculated value: C 45.16, H 6.50, N 12.54 Actual value: C 45.55, H 6.65, N 12.49 (iv) 2- (2-acetamido-2-deoxy-6
-0-glycyl-D-glucose-3-0-yl)-D-propionyl-L-alanyl-D-
Isoglutamine (82.4 mg, 0.15 mmol) in N,
Dissolve in N-dimethylformamide (1.2 ml),
Add N-ethylmorpholine (19.2μ) while cooling on ice. Add 3-(2,3-dimethoxy-5-methyl-1,4-benzoquinone-6-) to this solution.
Add p-nitrophenyl)-propionic acid ester and allow to react at room temperature for 16 hours. This was purified in the same manner as in Example 3 (ii), and 2-{2
-acetamido-2-deoxy-6-0-[3
-(2,3-dimethoxy-5-methyl-1,4
-benzoquinone-6-yl)-propionyl-
Glycyl]-D-glucose-3-0-yl}
- 79.0 mg of D-propionyl-L-alanyl-D-isoglutamine is obtained. [α] ³⁰ _D +33.3゜(c
0.5, ethanol) Rf ² = 0.62, Rf ³ = 0.29 Elemental analysis value C ₃₃ H ₄₇ O ₁₇ N ₅・H ₂ O Calculated value: C 49.31, H 6.15, N 8.71 Actual value: C 49.20, H 6.27, N 8.63 Example 15 (i) t-Butyloxycarbonyl-α-aminoisobutyric acid (0.96g, 4.7mmol) and HONB (1.00g,
Dissolve 5.6 mmol) in acetonitrile (10 ml), add N,N'-dicyclohexylcarbodiimide (1.16 g, 5.6 mmol) under ice cooling, and stir at 4°C.
Incubate for 16 hours. The precipitated crystals were removed and the solvent was distilled off under reduced pressure. The residue was dissolved in 10 ml of N,N-dimethylformamide solution containing D-isoglutamine benzyl ester (1.11 g, 47 mmol).
and incubate for 16 hours at room temperature. The solvent was evaporated and the residue was dissolved in ethyl acetate.
Wash sequentially with 1N hydrochloric acid, 5% sodium bicarbonate solution, and saturated saline,
Dry the organic layer with anhydrous sodium sulfate. Solvent evaporation. The residue was recrystallized from ethyl acetate-diethyl ether to give t-butyloxycarbonyl-α.
1.39 g of crystals of -aminoisobutyroyl-D-isoglutamine benzyl ester were obtained. melting point
112-113℃ t-Butyloxycarbonyl-α-aminoisobutyroyl-D-isoglutamine benzyl ester (632mg, 1.5mmol)
was dissolved in trifluoroacetic acid (7 ml) and stirred at room temperature.
Incubate for 30 minutes. The solvent was distilled off and ether was added to the residue to obtain α-aminoisobutyroyl-D.
-Remove the precipitate of isoglutamine benzyl ester trifluoroacetate. Dissolve this in tetrahydrofuran (10 ml) and neutralize by adding triethylamine (0.21 ml) under ice cooling to obtain a tetrahydrofuran solution of α-aminoisobutyroyl-D-isoglutamine benzyl ester. (ii) Add the solution of α-aminoisobutyroyl-D-isoglutamine benzyl ester obtained in (i) above to the benzyl 2-acetamido-4,6-0-benzylidene obtained in Example 11 (ii). 3-0
-(D-1-carboxyethyl)-2-deoxy-α-D-glucopyranoside HONB active ester (949 mg, 1.5 mmol) was mixed with 10 ml of tetrahydrofuran solution and mixed at room temperature for 16 hours.
Allow time to react. The reaction solution was concentrated, the resulting precipitate was collected, and benzyl 2-(benzyl 2
-acetamide-4,6-0-benzylidene-
2deoxy-α-glucopyranoside-3-0-
457 mg of D-propionyl-D-propionyl-α-aminoisobutyroyl-D-isoglutaminate was obtained. ^Rf1 =0.70. Elemental analysis value C ₄₁ H ₅₀ O ₁₁ N ₄ Calculated value: C 63.55, H 6.51, N 7.23 Actual value: C 63.82, H 6,88, N 6.87 (iii) Benzyl 2-(benzyl 2-acetamide-4, 6-0-benzylidene-2-deoxy-α-D-glucopyranoside-3-0-yl)
-D-propionyl-α-aminoisobutyroyl-D-isoglutaminate (973mg,
Dissolve 1.26 mmol) in 42 ml of 60% acetic acid and heat on a boiling water bath for 30 minutes. After the reaction, the solvent was distilled off and the residue was dissolved in ethyl acetate. Diethyl ether was added to this to precipitate it, and benzyl 2-(benzyl, 2-acetamido-2-deoxy-α-D-glucopyranoside-3-0-
505 mg of D-propionyl-D-propionyl-α-aminoisobutyroyl-D-isoglutaminate was obtained. [α] ²³ _D +9.8゜(c 1.0, ethanol) Rf=
0.46 [Chloroform-acetone-methanol (10:3:2), silica gel plate] Elemental analysis value C ₃₄ H ₄₆ O ₁₁ N ₄ · H ₂ O Calculated value: C 57.64, H 6.87, N 7.95 Actual value: C 58.07 , H 6.77, N 7.89 (iv) Benzyl 2-(benzyl 2-acetamido-2-deoxy-α-D-glucopyranoside-3-0-yl)-D-propionyl-α-aminoisobutyroyl-D-isoglutami nate (412 mg, 0.6 mmol) and carbobenzoxy-glycine P-nitrophenyl ester (396 mg,
1.2 mmol) in N,N-dimethylformamide (3 ml) in the presence of 1-hydroxybenztriazole (324 mg, 2.4 mmol) and N-ethylmorpholine (0.31 ml, 2.4 mmol) as in Example 14 (ii). After reaction at room temperature for 40 hours, purification was performed to obtain benzyl 2 [benzyl 2-acetamide-6-0-
356 mg of (carbobenzoxy-glycyl)-2-deoxy-α-D-glucopyranoside-3-0-yl]-D-propionyl-α-aminoisobutyroyl-D-isoglutaminate is obtained.
Melting point: 67°C (decomposition). [α] ²³ _D +76.8° (c 0.5, N,
N-dimethylformamide). ^Rf1 =0.54. Elemental analysis value C ₄₄ H ₅₅ O ₁₄ N ₅ Calculated value: C 60.19, H 6.32, N 7.98 Actual value: C 60.25, H 6.25, N 7.95 (v) Benzyl 2-[benzyl 2-acetamide-6-0- (carbobenzoxy-glycyl)
-2-deoxy-α-D-glucopyranoside-
3-0-yl]-D-propionyl-α-aminoisobutyroyl-D-isoglutaminate (307 mg, 0.55 mmol) was hydrogenated in the presence of palladium black in acetic acid in the same manner as in (ii) of Example 1. After that, it was purified to give 2-(2-acetamido-2-deoxy-6-0-glycyl-D-glucose-
3-0-yl]-D-propionyl-α-aminoisobutyroyl-D-isoglutamine 184mg
get. Melting point: 128℃ (decomposition). [α] ²³ _D +61.0゜(c
0.5,N,N-dimethylformamide). Rf ²
=0.44. Elemental analysis value C ₂₂ H ₃₇ O ₁₂ N ₅・1/2H ₂ O Calculated value: C 46.15, H 6.69, N 12.23 Actual value: C 46.26, H 6.93, N 12.29 (vi) 2-(2-acetamide-2 -deoxy-6
-0-glycyl-D-glucose-3-0-yl)-D-propionyl-α-amino isobutyroyl-D-isoglutamine (56.4 mg,
Dissolve 0.1 mmol) in N,N-dimethylformamide (1 ml), and add N-ethylmorpholine (12.8μ) under ice-cooling. Add 10- to this solution.
(2,3-dimethoxy-5-methyl-1,4-
Add benzoquinone-6-yl)-decanoic acid p-nitrophenyl ester (47.4 mg, 0.1 mmol), stir at room temperature for 16 hours, and purify in the same manner as in Example (ii) to obtain 2-{2- Acetamide
2-deoxy-6-0-[10-(2,3-dimethoxy-5-methyl-1,4-benzoquinone-
50.5 mg of 6-yl)-decanoyl-glycyl]-D-glucose-3-0-yl}-D-propionyl-α-aminoisobutyroyl-D-isoglutamine is obtained. [α] ²³ _D +30.6゜ (c 0.5, ethanol) Rf ² = 0.79 Rf ³ = 0.61 Elemental analysis value C ₄₂ H ₆₅ O ₁₇ N ₅・3H ₂ O Calculated value: C 52.21, H 7.37, N 7.25 Actual values: C 52.52, H 7.20, N 7.34 Example 16 (i) 11-aminoundecanoic acid (30.2g, 0.15mol)
Dissolve in 2N aqueous sodium hydroxide solution (500ml) by slightly heating (50℃). Carbobenzoxy chloride (30.7 g,
0.18 mol) was added in 4 portions over 30 minutes. Stir for another 2 hours. Cool the reaction solution on ice,
Adjust the pH to 2 with 6N hydrochloric acid and extract with ethyl acetate (600ml). The ethyl acetate layer was washed with water and dried over anhydrous sodium sulfate. The solvent was distilled off, petroleum ether was added to the crystalline residue, and after cooling, this was collected.
Carbobenzoxy-11aminoundecanoic acid
Obtain 63.0g. Melting point 96-97℃. ^Rf1 =0.67. Elemental analysis value as C ₁₉ H ₂₉ O ₄ N Calculated value: C 68.03, H 8.71, N 4.18 Actual value: C 68.05, H 8.73, N 3.78 (ii) Carbobenzoxy-11-aminoundecanoic acid (10.1g, 30mmol ) and p-nitrophenol (4.59 g, 33 mmol) were dissolved in a mixed solvent (120 ml) of ethyl acetate-N,N-dimethylformamide (5:1), and N,N'-dicyclohexylcarbodiimide (6.80 g, 33 mmol) was dissolved. In addition,
Stir at room temperature for 15 hours. The precipitate is removed and the solvent is distilled off. The residue is crystallized from petroleum ether. Ethyl acetate - Recrystallized from petroleum ether to give carbobenzoxy-11-
7.65 g of aminoundecanoic acid p-nitrophenyl ester is obtained. Melting point 72-73℃. ^Rf1 =0.94. Elemental analysis value C ₂₅ H ₃₂ O ₆ N ₂ Calculated value: C 65,77, H 7.07, N 6.14 Actual value: C 66.03, H 7.17, N 6,18 (iii) Benzyl 2-(benzyl 2-acetamide- 2-deoxy-α-D-glucopyranoside-3-0yl)-D-propionyl-L-alanyl-D-isoglutaminate (2.01g,
3 mmol) and carbobenzoxy-11-aminoundecanoic acid p-nitrophenyl ester (2.74 g, 6 mmol) in the presence of 1-hydroxybenztriazole (1.62 g, 12 mmol) and N-ethylmorpholine (1.54 ml, 12 mmol). Lower N, N
- After reacting in dimethylformamide (25 ml) at room temperature for 2 days, it was purified in the same manner as in (ii) of Example 3, and benzyl 2[benzyl 2-acetamide-6-0-(carbobenzoxy-11-amino)] undecanoyl)-2-deoxy-α-D
1.76 g of -glucopyranoside-3-0-yl]-D-propionyl-L-alanyl-D-isoglutaminate is obtained. Melting point 144-145℃. Rf ¹ =
0.66. [α] ²¹ _D +67.3° (c 0.5, N, N-dimethylformamide) Elemental analysis value C ₅₂ H ₇₁ O ₁₄ N ₅ Calculated value: C 63.07, H 7.23, N 7.07 Actual value: C 63.18, H 7.31, N 6.97 (iv) Benzyl 2-[benzyl 2-acetamido-6-0-(carbobenzoxy-11-aminoundecanoyl)-2-deoxy-α-D-
Glucopyranoside-3-0-yl]-D-propionyl-L-alanyl-D-isoglutaminate (1.70 g, 1.72 mmol) was hydrogenated in the presence of palladium black in acetic acid in the same manner as in (ii) of Example 1. After that, it was purified to give 2-[2-acetamide-6
-0-(11-aminoundecanoyl)-2-deoxy-D-glucose-3-0-yl]-D
-Propionyl-L-alanyl-D-isoglutamine 1.02g is obtained. Melting point: 120℃ (decomposition). ^Rf2 =
0.61. [α] ²¹ _D +20.7° (c 0.5, N, N-dimethylformamide). Elemental analysis value C ₃₀ H ₅₃ O ₁₂ N ₅・H ₂ O Calculated value: C 51.93, H 7.99, N 10.10 Actual value: C 51.85, H 8.00, N 9.82 (v) 2-[2-acetamide-6- 0-(11-aminoundecanoyl)-2-deoxy-D-glucose-3-0-yl]-D-propionyl-L-alanyl-D-isoglutamine (101.4
mg, 0.15 mmol) and 3-(2,3-dimethoxy-5-methyl-1,4-benzoquinon-6-yl)-propionic acid p-nitrophenyl ester (56.3 mg, 0.15 mmol) with N-ethylmorpholine. (0.03 ml) in N,N-dimethylformamide (1.5 ml) at room temperature for 60 hours. This was purified in the same manner as in Example 3 (ii) to obtain 2-{2-acetamido-2-deoxy-6-
0-[3-(2,3-dimethoxy-5-methyl-1,4-benzoquinon-6-yl)-propionyl-11-aminoundecanoyl]-D-glucose-3-0-yl}-D- 70 mg of propionyl-L-alanyl-D-isoglutamine is obtained. [α] ²³ _D +25.1° (after 3 minutes) → +25.4° (after 25 hours) [c 0.5, 70% ethanol]. ^Rf3 =0.43. Elemental analysis value C ₄₂ H ₆₅ O ₁₇ N ₅・H ₂ O Calculated value: C 54.24, H 7.26, N 7.53 Actual value: C 54.42, H 7.38, N 7.50 Example 17 (i) Methacrylic acid (1.72 g, 20 mmol) and N-hydroxysuccinimide (2.19 g, 19 mmol) were dissolved in 20 ml of acetonitrile and N,N'-dicyclohexylcarbodiimide (4.33 g,
21 mmol) and react at 0°C for 1 hour and at room temperature for 3 hours in the dark. The precipitate was filtered off, and the filtrate was distilled off under reduced pressure. Add petroleum ether to the residue, cool it, and filter the crystals to obtain crystals of methacrylic acid N-hydroxysuccinimide ester.
Get 2.5g. Melting point 100-102℃ Elemental analysis value C ₈ H ₉ O ₄ N Calculated value: C 52.45, H 4.95, N 7.65 Actual value: C 52.46, H 4.83, N 8.05 (ii) 6-0-β-alanyl-N -Acetylmuramyl-L-alanyl D-isoglutamine (564mg,
1 mmol) in N,N-dimethylformamide (10 ml), and add N-ethylmorpholine (0.128 ml) and hydroquinone (1 mg) under ice cooling. Furthermore, methacrylic acid and N-hydroxysuccinimide (238 mg, 1.3 mmol) were added to
After reacting for 24 hours at °C, the solvent was distilled off. Ethyl ether was added to the residue to obtain a powder, which was reprecipitated from ethanol-diethyl ether to give 2-[2-acetamido-2-deoxy-
6-0-(methacryloyl-β-alanyl)-
D-glucose-3-0-yl]-D-propionyl-L-alanyl-D-isoglutamine [hereinafter referred to as 6-0-(methacryloyl-β-
555 mg of alanyl)-N-acetylmuramyl-L-alanyl-D-isoglutamine was obtained. Melting point 164℃ (decomposition). [α] ²⁴ _D +36.3° (after 3 minutes)
→ +32.6° (25 hours later) [c 0.405, water]. ^Rf3 =
0.29 Elemental analysis value C ₂₆ H ₄₁ N ₅ O ₁₃・3/2H ₂ O Calculated value: C 47.41, H 6.73, N 10.63 Actual value: C 47.34, H 6.74, N 10.64 Example 18 2-{2-acetamide -2-deoxy-6-
0-[10-(2,3-dimethoxy-5-methyl-
2 mg of 1,4-benzoquinone-6-yl)-decanoyl-L-leucyl]-D-glucose-3-0-yl}-D-propionyl-L-seryl-D-isoglutamine was treated with 10 μg of squalene. Phosphate buffered saline containing 0.2% Twin 80 1
ml or 1 ml of physiological saline containing 0.2% Twin 80 to form an oil-in-water suspension. It is best to give 0.5 ml per individual for injection. Example 19 2-[2-acetamido-2-deoxy-6-
0-(retinoyl-β-alanyl)-D-glucose-3-0-yl]-D-propionyl-L-
Dissolve 8 mg of alanyl-D-isoglutamine in 3.5 ml of phosphate buffered saline and sterilize. This sterilized hydrogenated vegetable triglyceride (miglyol 812) - mannitol monooleate (17:
Add dropwise to 0.5 ml of the mixture of 3) while stirring vigorously to obtain an oil-in-water suspension. It is best to give 0.5 ml per individual for injection. Example 20 2-{2-acetamido-2-deoxy-6-
0-[3-(2,3-dimethoxy-5-methyl-
1,4-Benzoquinone-6-yl)-propionyl-L-prolyl]-D-glucose-3-0-
Il}-D-propionyl-L-alanyl-D-
Dissolve 500 mg of isoglutamine and 5 g of mannitol in distilled water to make 1000 ml. After sterilization, dispense 2 ml into vials and freeze-dry. Before use, this drug is dissolved in physiological saline and made into an injection solution. Reference example 6-0-(methacryloyl-β-alanyl)-N-acetylmuramyl- obtained in Example 17
L-alanyl-D-isoglutamine (530mg,
0.84 mmol) in N,N-dimethylformamide (2 ml) and reacted at 60°C for 15 hours under nitrogen atmosphere using α,α'-azobisisobutyronitrile (2.7 mg, 0.016 mmol) as a catalyst. The reaction mixture was poured into methanol (50ml) and an oil separated. The methanol was decanted and the residue obtained by distilling the methanol off under reduced pressure was dissolved in N,N-dimethylformamide, and chloroform was added to remove the precipitate. This was re-precipitated from methanol-chloroform and 6-0-(methacryloyl-
125 mg of a polymer of β-alanyl)-N-acetylmuramyl-L-alanyl-D-isoglutamine was obtained. Molecular weight measured by vapor pressure osmosis method: approximately 2000
Melting point 154-155℃ [α] ²⁴ _D +35.5° (after 3 minutes) → +
34.8° (25 hours later) [c 0.29, water] Rf ² = 0.00 Rf ³
=0.00 Elemental analysis value (C ₂₆ H ₄₁ O ₁₃ N ₅・3H ₂ O) as n Calculated value: C 45.54, H 6.91, N 10.21 Actual value: C 45.47, H 6.25, N 10.39 On the other hand, the above oily substance was , N-dimethylformamide, methanol was added, the solvent was decanted, and the residue was crystallized with ethyl ether to give 6-0-(methacryloyl-β-alanyl)-N-acetylmuramyl-L- Alanil-D
-145 mg of isoglutamine polymer is obtained. Molecular weight 4000 or more as measured by vapor pressure osmosis method,
Melting point 260℃ or higher, [α] ²⁴ _D +33.4° (after 3 minutes) → +31.2
° (25 hours later) (c 0.455, water) Rf ² = 0.00 Rf ³ =
0.00 Elemental analysis value (C ₂₆ H ₄₁ O ₁₃ N ₁₅・1/2H ₂ O) Calculated value: C 48.74, H 6.61, N 10.93 Actual value: C 48.64, H 7.12, N 10.07

Claims (1)

[Claims] 1. General formula [In the formula, R is a lower alkyl group, R ¹ is a carbon number of 3
~41 alkyl or alkenyl group, formula (In the formula, R ¹¹ represents an alkylene group having 2 to 9 carbon atoms.) or a group represented by the formula In the group represented by, n is zero or an integer of 1 to 9, ^R2 is a hydrogen atom or a lower alkyl group when n is zero, and a hydrogen atom when n is an integer of 1 to 9; is the formula R ³ is a lower alkyl group, R ⁴ is methyl or hydroxymethyl, R ⁵ is a hydrogen atom or methyl, R ⁶ is carbamoyl, and R ⁷ is a carboxyl group. show. ] A compound represented by or a salt thereof.