JPS5815991A - Glucosamine derivative - Google Patents

Abstract

NEW MATERIAL:A compound shown by the formulaI(R is <=6C organic acid residue; n is 11-34; R<1> is H or lower alkyl; R<2> and R<3> are H or lower alkyl which may be replaced with OH; R<4> is H or lower alkyl; R<5> is carboxyl which may be esterified or amidated; D and L are in steric configuration when carbon atom is asymmetric) or its salt. EXAMPLE:2{2-Acetamide-2-deoxy-6-0-[16-( 2, 3-dimethoxy-5-methyl-1, 4-benzoquinone-6-yl)hexadecanoyl]-D-glucopyranose-3-0-yl}-D-propionyl- L -valyl-D-isoglutamine-methyl ester. USE:An immune function enhancer. PROCESS:A compound shown by the formula II (R<2>' and R<3>' are H or lower alkyl; R<5>' is protected carboxyl, etc.) is condensed with a compound shown by the formula III, and, if necessary, deprotected, to give a compound shown by the formulaI.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel glucosamine derivatives with excellent biological activity. More specifically, the present invention relates to a general formula having an excellent immune function-enhancing effect [wherein R is an organic acid residue having 6 or less carbon atoms, and n is an integer of 11 to 34, R is a hydrogen atom or a lower alkyl V and R3 each represent a hydrogen atom, 9 represents a lower alkyl group which may be substituted with a hydroxyl group, P represents a hydrogen atom or a lower alkyl group, and Rs represents a carboxyl group which may be esterified or amidated. y
py group, and (D) and (L) each indicate the steric configuration when the base atoms are asymmetric, and salts thereof are provided.

Regarding the above general formula (1), R is preferably acetyl, propionyl, butyry~, isobutyry~, parelli μ, etc., having 6 or less carbon atoms. RlHa R3 and R
The lower alkyl group as 4 may be either linear or monobranched, preferably having 61 carbon atoms, and 9 is methyl, ethyl, n-propyl, isopropyl; Buchi ~, bench ~, hexy μ that is branched at any position
Among them, those having up to 4 carbon atoms are preferred.

The force μ-box V group as R8 may be esterified with lower alkyl as R2 to R5, or substituted with ammonia or one or two of the lower alkyl groups and amidated with 9 amines. good.

Among the compounds of the above general formula (1), compounds in which R is an acetiμ group and R is a methoxy group are particularly preferred. Further, when HL4 is a hydrogen atom, R3 is preferably a lower acetate group or a todorokiVmethyμ group.

In the above compound (1), the gμ cosamine residue is preferably in the D type, and the R1-substituted acetic acid residue bonded to the oxygen atom at the 3rd position is in the D type when R represents a lower alkyl group. ,
Also R2. R3 substituted amino acid A/residue, R''*
When both R and hydrogen atoms are not the same lower atom group, the L-type is preferred, and the aminoaminocyanic acid is preferably the D-type isoglutamic acid.

Compound (1) is either acidic or neutral depending on the type of substituent, and when it is acidic, it forms a salt with a base. Salts with bases include, for example, ammonium, salts with alkali metals or alkaline earth metals, such as salts with sodium, potassium, sium, and magnesium. Compound (1) can be used as a physiologically acceptable salt thereof.

The compounds obtained by the present invention have low toxicity and exhibit excellent immune function-enhancing effects, particularly enhancing cell-mediated immune responses, which are important for protecting living organisms from infection. In addition, this compound
It is clear from the experiments described later that it also acts on biological membranes and exhibits a destabilizing effect on linseam membranes.

The following experiment shows that the present compound enhances the cellular immune response, which is important for the protection of living organisms against infection, among other immune-enhancing effects.

The enhancing effect of this compound on cell-mediated immune responses in living organisms can be seen, for example, in Mofi/Mott's N-acety-tin-3-azobenzene-4'-arsonic acid (ABA-[-A
c-Tyr) It is shown by its immune-enhancing effect on induction of delayed-type hypersensitivity to Ac. That is, 5 Gμg per mu B mu summer-mu C-tyrc) and the present compound (
I got one! DIO-100 μg) was dissolved in phosphate-buffered saline and mixed with Freund's incomplete adjuvant (
Hartley guinea pigs are immunized by injecting 0.05 LIt of each tailing with a water-in-oil suspension prepared by mixing with F.M. Hair removal from the back for 2 weeks, AB Mu Thin 1
-A-amylase (100 μg) was injected 2 times
After 4 and 48 hours, the diameter of the skin reaction (erythema and induration) is measured. Table 1 shows the results obtained using the compounds of the present invention, and the diameter can be seen as a measure of cell-mediated immunity.

On the other hand, even if the antigen ABA-N-Ac-Tyr was dissolved in phosphate buffered saline and mixed with Freund's incomplete adjuvant and a water-in-oil suspension was administered, muBA - No delayed hypersensitivity to N-Ac-Tyr is observed. As described above, the metal article of the present invention has a strong immunoenhancing activity against cell-mediated immune reaction (delayed type hypersensitivity) K.

It is especially effective when administered to suppress carcinogenesis when the carcinogenic rate increases due to aging (for example, due to aging) or when the carcinogenesis is likely to occur in mammals, especially warm-blooded animals. They can be administered enterally, eg orally or rectally, or parenterally. Its dosage depends on the individual condition of the warm-blooded animal.

Depending on the age of the species and the dosage form of WMK, isotonic solutions, e.g. When administered by subcutaneous, intradermal, or intramuscular injection, the preferred dosage is about 1 to 500 μg/kg/11, particularly about 5 to 30 μg/kg/a of the compound (1. in anhydrous form).

When administered by injection, a water-oil stable emulsion, preferably an oil of vegetable or animal origin, may also be administered. For example, the amount of vegetable oil used is approximately 5-100 volumes of isotonic aqueous solution, the amount of vegetable oil that can be metabolized is approximately 5-100 volumes of animal oil, and the amount of animal oil is approximately 5-100 volumes.
It consists of a nectar fixative. For administration via the buccal route, it can be administered, for example, as a sugar-coated tablet, with other excipients that can be used in the formulation, in which case the dosage is about 40-4000 pgAt<7
Within days.

The compound (1) of the present invention has nine general formulas [wherein R,
R'', R', (D) and (L) have the same meanings as above, and R' and R' each represent a lower atom group in which 9 hydrogen atoms are substituted with protected hydroxyl groups; The base,
Rs' represents a Vμ group which may be protected or esterified or amidated, and V represents a protected hydroxyl group] and the compound represented by the general formula [where n is the same as above] (K) is condensed with a compound represented by
can be manufactured.

Regarding the above general formula (seal), "' e "' @ R't
As the protecting group for the hydroxyl group contained in the group represented by ' and the force μ boxy μ group represented by R', any known easily removable protecting group may be used. For example, an ay group (for example, a tertiary butyN group), i9 is a tetrahydrobionyl group, nine is a bencyiiv*μbonyl group, and nine is a lower alkanoylcarbonyl group, and so on. A benzyl group optionally substituted with a lower alkoxy group is conveniently placed in front of the halogen atom, nitro group, or lower a[mu]kyl group. Examples of the protecting group for the .mu. group include an alkyl .mu. group (such as a tertiary .rho. group) or a benzyl group.

The condensation of the compound (1) and the metal compound (1) can be carried out using a condensation method known per se.

For example, it is preferable to activate the Vfi/group of a metal compound and react with the compound (1).

Examples of activated running force μ-boki\N groups include active Esthe μ and acid anhydrides.

As active esthetics, p-nide p-phenylene ester,
■- and droxysuccinimide ester μ, p-hydropki V-5-noμbonene-2,3-diiI voice boximide ester, front is unsubstituted or halogen group, hedagenated methyl voice group Examples include 1- and dolo-benzotriacetic μ esters substituted with meth-F-y)v groups.

Preferred acid anhydrides include mixed acid anhydrides and acid amides such as imidazolide and inxacilide.

In the reaction of activated nine esters, organic bases such as triethyamine, summer-methylmorpholine, M-
It is also possible to coexist with ethylA/mophorin and 1-w droxybenzo).

Activation of force〃boxyμ is an escape compound (seal) and N, 'H'
It can also be carried out by directly reacting -dicyclohexyl Vlv force with positive imide.

The reaction temperature is usually about 0°C to about 80°C, preferably about 5°C to 50°C. The reaction may be carried out at other temperatures if desired.

The reaction generally proceeds in a solvent, and examples of the solvent include ethers such as tetrahydrofuran and dioxane, ethers such as ethyl acetate and isoamino acetate,
9 For example, N, Seal-Dimethylformamide page 0-Methi*-
2-Birylidone, W, M-dimethyμ acetamide, etc. Summer-ap! Appropriate solvents such as fμ amides, pyridine, dimethyp sulfoxide, hexamethylene phosphoamide, etc. may be used.

The above compound (1) reacts with compound (1) in a duri-stoichiometric manner, but it goes without saying that these two components do not necessarily have to exist in an equivalent relationship in the dihydric system; I) Compounds (1) and 19 are used in amounts of about 1 to 57, preferably about 1-/2 mo.mu. of their active derivatives. The quantitative ratio of the two components can be appropriately selected to give good results depending on the combination of raw materials and other conditions.

Beforehand, the unreacted raw material can be appropriately recovered and used again as a raw material.

After the above reaction, the protecting group can be removed by a method known per se. For example, by hydrogenation in the presence of a noble metal catalyst (e.g. platinum or badge rum)
can be hydrolytically removed by acid hydrolysis.

The thus produced metal compound (1) can be processed by known processing means (for example, extraction, dissolution, various chromatography,
It can be isolated in a free state or as a salt as described above using crystallization, recrystallization, reprecipitation *fk).

Note that the raw material metals (1) and (seals) can be manufactured using known compounds as raw materials by means of expanding methods known per se or by a combination thereof, and examples of the manufacturing methods are described below as reference examples. . All of the intermediate metals listed in the reference steel along with the physicochemical data are new metals.

The present invention will be explained in more detail below using Examples, but it goes without saying that these are not intended to limit the present invention.

Example 1 (I) 2-(bendiμ 2-acetamido-2-deoxyS' -a -D-g-kopitunoVd-3-〇-yl)
-D-Poupioniμ-L-BaryluD-i')l'Wl
Mi￥ )flWXZffi/(1fJT*, 0°3m
mol) and 16-(2,3-dimethoxyF-5-methyl-1,4-benzoquinone-6-yA/)hequinadecanoic acid, I-dimethymuformamide (hereinafter abbreviated as DMF) C
2d)-Viridi7C0, 5d) mixture of KK” melted,
Add ditalohex Vlv force μ positive imide (hereinafter abbreviated as DCC) (124jIF, 0°@mmol). A 4g reaction was carried out at room temperature.

The precipitate was removed, the *removal was distilled off, and the residue was purified by V9iI gel~
Purify using a preparative thin layer CLIMA F-GUTSUFI (Ws solvent: methanol-chloroform 5:95).

Scrape off the target part and add methanol-chloroform (
Extract with a mixed solvent of 4:6) and evaporate the solvent. The residue is dissolved in chloroform, washed with water and dried over sodium sulfate. The solvent was distilled off, and the residue was solidified with ether to give 2-[benziμ 2-acetamido-2-deoxyS/-
6-0-(16-(2,3-dimethoxyV-5-methyl-1,4-benzoquinone-6-yA/)
]-α-D-gμcopifluorVdo3-0-iμ]-ri1deropioni/M-L-va1μm D-isoglutamine methyl ester (189111J, 60.4%) is obtained. Melting point 188-180°C. (tt) D + 59.2
”(00,5,DM!”).

Elemental analysis value 0.4H,,0□61I4・114H! o
Calculated value: CB1.83. H7,95,M
5.33 actual measurement value: C61,45,H7,93,M
5.51(1) 2-(bendifi/2-acetamide-
2-deoxy$/1-0-(1g-(2,3-dimethoxys)
/-5-methyl-1,4-benzoquinone-6-iA/)
heki fdecanoi μ)-g-D-glucopifusito-3-
0-i~)-D-propioni*-L-bashiμ-D-
Isogp Tine Methiμ Esthe 1v (17G111.0
．． 163 mmol) was subjected to catalytic reduction in acetic acid <set> using badge rum black as a catalyst at room temperature for 4 hours. The catalyst is separated from the furnace and the solvent is distilled off.

Dissolve the residue in methanol (5d) and dissolve the second chain chloride 6
Hydrate (27011, 1m11o1) was mixed with water (0°3g
Add the solution dissolved in g) and allow to react at room temperature for 30 minutes.

Water and chloroform are added to the reaction solution for extraction. The five layers are washed with water and then dried with sodium sulfate. The solvent was distilled off, and the residue was purified by micro-preparative thin layer taromatophyll (solvent: chloroform-acetic acid-water-90:10:
Purify in step 5). Scrape off a portion of the target product and extract with a mixed solvent of methanol-chloroform (3N). Prepare a solvent, dissolve the residue in chloroform (20mK),
Wash with water, sodium sulfate 9? Dry in step 5. *M was distilled off, and the residue was reprecipitated from polypropylene chloride-petroleum ether to give 2-[2-acetamido-2-deoxy-6-Q-
(16-(2,3-dimethoxy-5-methyfi/-1,
4-benzoquinone-6-yp-/)hexadecanoyy)
-D-gμcobifunose-5-o-iμ)-D-propionyl-L-pariμ-D-isoglutamine methyl ester ~ (105q, 87.6%) is obtained. Melting point 193-1
95℃. (&)%)'433.1' (o O-5e
Ethanol) Elemental analysis value c4. Calculated value as it, 6y, o□6・it, O: C” 58.13. H8,10,M
5. '78 dormitory measurements: C58, 33, III 8.1
4. MS, 81 Example (1) 2-[benzi-2-acetamido-2-deoxyFa-a-D-gu]
Kopitsuno V Do 3-0-I μ)-D-Propioni μ-L
-9fil-D-isoglutamine methyl ester and 22-(2,3-dimethoxyV-5-methyA/-1,4-
Using benzoquinone-6-iμ) doconanoic acid, 2-[bendi/l/2-1cetamido-2-deoxyS/-1! -0
-(22-(2,3-s/methoxyV-5-methoxy14/-
1,4-benzoquinone-6-i~) docosanoid μ)-m
-D-gμ cobilanoside 3-0-iμ)-D-propioniμ-L-valyl D-isoglutamine methyl ester (27.8%) is obtained. Melting point: 182,184°C.

(ri%”+52.8° (c O, 5, DMr).

Elemental analysis value C5oHeaox6Na・”Calculated value as AH20: C63,41,H8,42,M 4.9
Measurements for 3 dormitories: C63.89, H8.53, N 5.2
1(1) Example/- In the same manner as in (1), 2-[2-
Acetamide-2-deoxy-8-0-(22-(2,
3-dimethoxy\-5-ichi~-1,4-benzoquinone-
6-i μ) Docosanoid μ〃-D-g〃Kobilanose-3
-0-i~)-D-propioni~-L-paraμmD-isoglutaine methyl ester xy* (48.2%) is obtained.

Melting point: 204°C.

(J B +28. $11 (c O-5m ethanol-/I/).

Elemental analysis value Calculated value as casHsa page, O engineering, II, O: C6G, 32. H11,60,M5.
31 actual measurements: C60, 42, H8, 65, N 5.
35 dormitory Example 3 (1) In the same manner as in Example/-(1), 2-[bendiA'2-acetamido 2-deoxyV-a-D-glucobionoside 3-0-iμ)-D -De p Pioni μ-L-
Alanil-D-Isoglutamine Benji μ Esthe~ and 2
2-(2,3-dimethoxyS/-5-methoxyA/-1,4
-benzoquinone-6-i) 2-[bendi] 2-acetamido-2-deoxy$/-1i-0 using toconic acid
-(22-(2,3-dimethoxyV-5-methoxyμm1.4
-benzoquinone-6-iA/)-do5'']-zanoiμ)
-g-])-]g〃cobylanozidode-3-0-yl-D-
Obtain D-Isoglutamine A'-L-7Tuni/l/-D-Isoglutamine Benzi μEste* (11.3%). melting point 16
2-164℃. (g)'%F + 64-3° ((1G
,5. DMr).

Elemental analysis value CmaHeaMaozs 11! : L?
Calculated value: C65, f19. HIl, C6,I
4.77 Actual 11 value: C65, 26, HL21. M
4.7! [Seal] Example/-2- in the same way as (1)
[2-acetamido-2-deoxy-6-0-(22-
(2,3-dimethoxyV-5-methyμm1.4-benzoquinone-6-i, A/)+:'gosanoy]-D-darcopifnose-3-0-i~)-D-ppioni~- L-
Afni-D-isogutain (8G, 2X) is obtained as a lyophilized product from t-butano-μ. (a)v+
27-2° (co-5*ethano-A/).

Elemental analysis value Calculated value as CBOH81N40□6”BtO: C59,25, H8,35, MS, 53 Actual value: C5B, Go, H8, 48, pages 5, 26
Example 3 4-(2,3-dimethoxy-5-methyl-1,4-benzoquinone-6-yA/)tetratriacofonic acid (1
1011F, 0.16 mmol) and 2-[2-acetamido-2-deoxyS/-D-glucobifunose-3-
0-yl)-D-propioniμ-L-valyl D-isoglutamine methylester #(85,6”F, 0.
16 mmol) J>j and p-) Luens N fonic acid (1
1F) was dissolved in a mixed solvent of pyridine (3sg)-dioxane (1d), and DCC (66'1.0.32mmol
) is added. React for 90 hours at room temperature. The precipitate is evaporated and the solvent is distilled off. The residue was purified by silica gel preparative thin layer chromatography (extracted with M solution: chloroform-acetone-
Purify with methanol-*-10:3:2). Extract the target part with a mixed solvent of the same composition as the developing solvent. The solvent was distilled off, the residue was dissolved in chloroform and washed with water.
Dry with 9 um of sulfuric acid. The solvent was evaporated and the residue was reprecipitated from chloroform-petroleum ether to give 2-[2-acetamido-2-deoxy-6-0-(34-(2,3
-dime[xy-5-methyμm1.4-benzoquinone-6
-I) Tetratriacontanoy~) -D-gμcobylanose-a-O-iμ)-D-propioni~-ri-variμ-D-ingμtine methyl ester (16 cape, 8
．． 3%). Melting point 198-200℃, t:a, +
, +25.6° (cO,3,Ethano-A/).

Elemental analysis value C6t engineering 11"40□6・Hl, calculated as 0 fi: C63,8G, H9,39,N 4.
5m1 dormitory [1F: c s3.44. H9,42,
N 4.30 Reference Example (1) 22-(2- and droxy-3,4-dimethoxy-6-methylphenyfi/)tocosonic acid (3,04 g
e&! 11101) to acetic anhydride (3sr)Ki
ml! 1 and sulfuric acid (0.5 wt), and reacted at room temperature for 1 hour. Acetic anhydride was distilled off, and the residue was dioxane (2
0IIt) and 5% aqueous sodium hydrogen oxide (5Gm) were added, and the mixture was stirred at room temperature for 15 hours. Methiacetic acid (50 d) was added for extraction, and the organic layer was further washed with 1N hydrochloric acid and water, and dried over 9 ml of sulfuric acid. WI rope is distilled off and oily 22-
(2-acetoki$'-3,4-dimethoxyS/-6-methymuphenymu)tocosanoic acid (2.80 g) is obtained. Dissolve this in benzene (5m) and 9L of oxa/chloride (1
, 2 mg) and reacted at 50°C for 1 hour. The solvent was distilled off, toluene was added, and the solvent was further distilled off under reduced pressure.

Add sodium hydroxide, dry thoroughly in a desiccator, and 22-(2-acetoxy-3,4-dimethoxy-6-
Methylphenyl/%/) tocosanoic acid chloride (2.

68 g, 100%). Nuclear magnetic resonance spectrum A/(
δ value in deuterated chloroform (same below): 1.20-
1.53 (3811, b, CH,), 2.35, 2.3
9 (611, s.

C0CH,, nuclear CHs), 2.95(4H,t,CH
, C, O, nuclear cu.

), 3.88.3.90 (SR, s, OCH3), 4.
75 (IE[.

S, nuclear H).

22-(2-acetoxy-3+ 4-dimethoxy-6-
Methypheni) tocosanoic acid chloride (268g, 4.
7 mmol) was dissolved in chloroform (2 m), and this was added to 1-morpholino-1-cyclododecene (1,
60g, 6. 11111101) and triethylamine (0.85 s).
Stir for an hour. Add G normal hydrochloric acid (3d) to the reaction solution,
Heat to reflux for an hour. After returning to room temperature, extract by adding chloroform and water. The chloroform layer was washed with water, dried over sodium sulfate, and the solvent was distilled off. residue (yv)
Purification was carried out using a sintering agent (60 g) as a carrier to obtain 2-[22-(2-acetoxy-3,4-dimethoxy-6-methoxypheniμ). Docosanoy/L/]cyclododecanone (2.50 g, melting point 7171°C) is obtained. This is ethanol-fi/(5sf)
Dissolve while heating K, heat 6N aqueous sodium hydroxide solution (3d
) is added. After refluxing for 15 minutes, the mixture was returned to room temperature and extracted with 1N hydrochloric acid (30s?) and chloroform (50wt). The chloroform layer was washed with water, dried over sodium sulfate, and the solvent was then distilled off. The residue was recrystallized from methiacetate to give 13-oxo-34-(2- and 3'-
3,4-dimethoxyV-6-methylphenymu)tetratriacontanoic acid (1,35 g, 32°rx) is obtained. Melting point 92-93°C.

Calculated value as elemental analysis 11043M, 606: C74,95,H11,12 Actual value:
C74,88,H11,2!1(1)13-oxo-3
4-(2-and doroki V-3,4-dimethoxy-6-methylpheny A/)tetratriacontanoic acid (1,33 g,
1.93 mmol), p-) ~ engineering sulfonic acid and tosyl t-dofudine (4554,2,5 mmol) in DM
Dissolved in a mixed solvent of F(5d)-sulfone (5d),
Heated to 1001°C and added KS/chlorohexane (10d
) and sodium yanoborohydride < 504sF,
9 mmol) and reacted at 100°C for 30 minutes. The temperature was returned to room temperature, hexane was distilled off, and water (50%
mg) and chloroform (50m) for extraction. After washing the chloroform layer with water and drying with sodium sulfate,
The solvent is distilled off. The residue was recrystallized twice from methoxy acetate to give 34-(2- and droxy-3,4-$7methoxy6
-methy~phenyl)? , toftriacontanoic acid (920
11F, To.

6%). Melting point: 93°C.

Elemental analysis value C4$H? ! Calculated value as 105: C
76, 5G, H11, 65 dormitory measurement: C76,37
, H11,91(1) 34-(2-hydroxyS'-
3,4-dimethoxy\-6-methyμpheniμ)-5:'tratriacontanoic acid (12011F, 1.36 mmol
) with DMF' (25 mg) and taaamk,um (50 mg).
S? ) of $alK, bis(salicylidene) ethylenediiminocobalt (π) (16511jF,
0. 51 mmol) and allowed to react at room temperature for 6 days. Chloroform (100d) and water (100I) were added to the reaction solution.
Add and extract d). The chloroform layer was washed with 1N hydrochloric acid and water, dried over 9 um of sulfuric acid, and the solvent was distilled off. The residue was purified using a Kahum chromatograph using silikage* (30 g). Develop with chloroform, collect fractions containing the target product, and evaporate the solvent. The remaining crystals were washed with hot methanol and 34-(2,3-dimethoxyV-5-methyfi/-1,
4-Benzoquinone-6-i~)tetophtriacontanoic acid (200ml, 21.3%) is obtained. Melting point 89-90℃
.

Elemental analysis value Calculated value as casa, aoa: Actual measurement value of C74,95,H11,12:
C74,3? , H11,091・-

Claims (1)

[Claims] General formula [wherein R is an organic acid residue having 6 or less carbon atoms, n is 11
-34 integer, V is a hydrogen atom, 9 is a lower alkyl group, V and P are each a hydrogen atom or a lower alkyl group optionally substituted with a hydroxyl group, R4 is a hydrogen atom or a lower alkyl group, Ha 9 indicates a carboxylic group which may be amidated, and φ) and French) respectively indicate the configuration when the hydrogen atom is asymmetric. salt.