JPS5877882A - Immunomodulating novel compound, manufacture and immunomodulator - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed Description of the Invention] The present invention is based on the general formula (■): ? (012) nOR' (1) represents a hydrogen atom or a town) Human being! xypurine residue, n is an integer of 1 to 6 or an a-amino acid, hydrogen atom, methyl group, P-human mixibenzyl group or aminopentyl group, and R3 is a hydrogen atom or an oligopeptide residue having 2 to 6 amino acids (where m is an integer of 2 to 6, R2 is the same as above, R' is OHi or 2
is an oligopeptide residue with ~6 amino acids)
and R4 are the same as above), and especially when two is 1, the general formula: R8 and R are the same as above) or the general formula: R8 and R is the same residue as above)
The compound represented by the formula <xb-a>: The method for producing the compound and the formula <xb-a> Contains at least one compound represented by the general formula (I) as an active ingredient. The present invention relates to a pharmaceutical composition having an immunomodulatory effect.

The "immunomodulating effect" of the present invention refers to an effect of interacting with cells having immune capacity and activating the immune capacity of the cells.

Recently, in the field of immunopharmacology, a series of extracted polypeptides with thymic hormonal action have been investigated, and they can be used as antigen supplements in clinical medicine, such as the treatment of several infectious diseases, especially viral infections or malignant abscesses. used in the field.

For example, a drug containing 12 different l-lipeptides extracted from bovine thymus is known. The drug is a virus,
The aim is to enhance the immunomodulatory effect on the attack of bacteria and even tumor cells. The amino acid sequence of the extracted final peptide has not yet been elucidated, and
It is also unclear what specific role the l/IJ peptide has in regulating immune action.

We have succeeded in synthesizing derivatives of components effective for the human immune system that have immunostimulatory effects against DNA or viruses in 11 people.

However, even in these cases, activation of the immune system appears to be nonspecific and lacks selectivity for one or more lymphocyte functions (e.g., stimulating or suppressing the immune response). ).

Similarly, activation of non-specific lymphocyte functions by the above-mentioned drugs is also uncertain.

Therefore, the main objective of the present invention is to provide a compound that has a specific immunomodulatory effect that can activate only a cell fraction that has a specific function.

According to the present invention, it has been found that the above object can be achieved by using a compound represented by the general formula (1), with the α-amino acid being the same or partially or completely different.

The a-amino acid can be used even if it is in the optically active d-form or hepta-amino acid, but the optically active mono-amino acid is preferred. The a-amino acids in the compound represented by the general formula (1) include glycine, alanine, valine, lysine, inlysine, serine, threonine, aspartic acid, glutamic acid, arginine, lysine, cysteine, cystine, methionine, phenylalanine, Examples include tyrosine, tryptophan, gastric phosphorus, and histidine-8.

The -1 amino acids are linked by typical peptide bonds in which the amino acids themselves are condensed with each other according to the previously formed sequence. That is, the amino group of one amino acid is bound to the carboxyl group of the other amino acid.

The hydroxypyrimidine ring of the purine derivative of the present invention may be an enol type as represented by general formula (1), a keto type, or a mixture of an enol type and a keto type.

The sequence and nature of the amino acid bound to the purine ring qualitatively changes the hormone-like message, whereby compound (1) chemically acts on the α-vacuole.

Therefore, by using a compound represented by the general formula fI), only a cell fraction having a specific function can be activated. Furthermore, compared to known analogous compounds, the compounds of the present invention can have more limited clinical use against the pathology of immune deficiencies, immune inactivations or autoimmune diseases, and malignant tumors themselves.

As mentioned above, the present invention relates to a method for producing a compound represented by general formula (1), and can be summarized as follows.

That is, general formula (1): %Formula %(1) Hydroxypurine residue represented by hydrogen atom or MH2, n is an integer from 1 to 6) 6
-Hydroxypurine and general formula (Rin): (In the formula, - is the same as above, R5 is a protecting group for amine 7 or a terminal amine commonly used in peptide synthesis) 2 to 6 groups whose groups are protected An oligopeptide residue of an a-amino acid in which the amino group represented by (a) is an oligopeptide residue having an amino acid is protected, and a known a-amino acid oligopeptide such as N,M'-carbonyldiimidazole, dicyctetrahexylcarbodiimide, and similar compounds thereof. A reaction is performed by activating the carboxyl group using a reagent to produce an intermediate represented by the general 2〇'IQ:R' (wherein P, n, R'' and R5 are the same as above), Then, the general formula (Im) consisting of removing the bond of the amino group: P -(OH,)n-
o -oo -am -NHR3(Ia)2 (in the formula, Ps n % R2 and R3 are the same as above); An intermediate represented by the general formula (7); General formula (Ib) consisting of activating the carboxyl group by known methods, protecting the carboxyl group if necessary and reacting with an acenoic acid or a peptide: (wherein ?, *, Peng, R2 and R4 is the same as above) and a method for producing a compound represented by the general formula 1) and phosgene (000
! 2), and then α-amino acid or general formula A(') An intermediate represented by the general formula @: (where PsnsR2 and R6 are the same as above) is prepared by reacting with an oligopeptide represented by This is a method for producing a compound represented by the general formula (Ia): (wherein PsnsR'' and R4 are the same as above), which comprises removing the group.

In addition, a compound represented by general formula (1) having a side chain different from that shown above is produced according to a production method similar to that described above.

Furthermore, the compound of the present invention or a corresponding intermediate can also be produced by using a similar compound having a chlorine atom instead of a hydroxyl group as a starting material and replacing the chlorine atom with a hydroxyl group by a known nucleophilic substitution reaction. The body is manufactured.

The starting material, a compound represented by the general formula (1): % formula % (1) (in the formula, n and P are the same as above), was prepared by Scheer H08, et al.
B.

at aj, ) (J, kh6.0h@m,,21
, (196B')). Next, the reaction formula is shown.

(O)I2)n-OH I (09n-1 OH The melting point and elemental analyzer of the hemisuccinate ester expressed by the general formula (■1) obtained by reacting with water succinic acid: % formula % (1) (in the formula, i and n are the same as above) were Shown in the table.

Reference compound (11) Wp+ 262°o1 Kondo et al.
aj ) (MakroI!1oj, Ohem, 29
8-501 (1969)) Next, the present invention will be explained with reference to Examples, but the present invention is not limited only to these Examples.

Example 1 [General formula (Vl-2μ(aH2)2-000-(OH
2) Production of 9-(2-hydroxyethyl)-6-hydroxypurine hemisuccinate represented by 2-000H(yl-2) (wherein - is the same as above) (4) Succinic anhydride 1 .24g (12,5mmole)
was dissolved in 20aj of warm pyridine, and to this was added 0.749 of 9-(2-hydroxyethyl)-6-hydroxypurine.
(4.1 mm + o1 m ) and a small amount of dimethylaminopyrimidine (hereinafter referred to as DMAP) using a spatyl was added, and the mixture was refluxed for 22 hours with stirring. Then, pyridine was distilled off, and the residue was washed with alcohol to remove excess co/monophosphate. After further filtration, the residue was dried to obtain the title compound (up 7210-2150).

Furthermore, the compound was analyzed by thin layer chromatography (hereinafter referred to as TL).
(referred to as O) (80% ethanol - 20% triethylamine
%), and the 41I structure was determined by engineering R spectrum analysis.

(to))? -(2-hydroxyethyl)-6-chloropurine 0.209 (1 mmole) and anhydrous amber #1
After pyridine was distilled off, the residue was dissolved in 10 ml of formic acid and refluxed for 50 minutes. After cooling, add 20 ml of methanol and filter the precipitate with suction to obtain the title compound.
I got it.

Example 2 - [General formula: P”-(aH2)2-0-00-(aH2
) Preparation of 6-hydroxy-9-(2-(2-chlorocarbonylethyl)-carbonyloxyethyl)-purine hydrochloride represented by 2-aool·Ho1 (in the formula, - is the same as above)] General formula 100 ml of hemifusuccinic acid ester represented by (Vl-2) was mixed with 5 ml of thionyl chloride at room temperature.
The mixture was dissolved in water and stirred at room temperature for 24 hours. Then, the excess acclimated thionyl was distilled off under reduced pressure, and the obtained white solid was dissolved in anhydrous ethyl ether and washed several times with ethyl ether using a suction P stand to obtain the compound shown in the table as white fine crystals. (Collapse temperature: over 200°Of) The structure of the compound converts the compound into a methyl ester (mp: 183~1ascb
) and determined by NMR and mass spectrometry.

Example 3 6-hydroxy-9-(2-(2-amino-6-(4-hydroxyphenyl)propionyloxy))-ethylpurine represented by (wherein - is the same as above) Production] (A) 0-benzyl-boa-tyrosine 571mp
(1 mmol 1 m) with anhydrous dimethyl sulfoxide (hereinafter referred to as
The mixture was dissolved in DMsO), added with carbonyl cylindazole is68mp (2 mmole), and heated at 70°C for 5 hours. The reaction mixture was then mixed with 9-(2-hydoxyethyl)-6- and 280,111 g (111
1! +01@) was added to the solution. 1:50 was added to the resulting reaction mixture. The separated oil was extracted twice with 25 aj of ethyl acetate, and the extracted product was dried and distilled under reduced pressure to remove ethyl acetate. The oily residue was dissolved in 20 mJ of ethanol without further purification and then dissolved in 5 ml of cyclohexane and palladium/
120 m9 of charfur was added and the mixture was refluxed for 2 hours while stirring. The precipitate that precipitates out during reflux is dissolved in acetic acid, and after the charcoal is separated, 4 to 5
m was added and refluxed for isO minutes. The acetic acid was then distilled off in vacuo, and the residue was washed with methanol to give the title compound as colorless crystals. (Yield: 87%, up 125
5-240qO) Next, the characteristic values of the title compound are shown.

IMI spectrum (in DM80, internal standard + TM8
)LOJ 211 (double@t) % -0
M4, Oa III (triplet) QH-
OH24,10J 4H (singlet) aH2
-OH24,6-7. DJ 4M (AA'ss')
(16H47°9 and 8-OJ 2H(aingl
etc) Purine hydrogen atom 11, OJ 4H (bro
ad bmd) variable hydrogen atom-)0-benzyl-M
-Boo-tyrosine 571 mp (1 m Peng 01.) dissolved in anhydrous acedinitrile, 9-(2-hydroxydiethyl)-6-crylobulin 198 mp, dicyclohexylcarbodiimide 210 mp and 4-bi4-ridinopyridine 25 mp was added and heated at 60°0 for 12 hours. After the resulting reaction mixture was cooled, the dicyclohexylurea was removed from door to door, and the solvent in the p solution was distilled off under reduced pressure. The residue was not purified further and was diluted with methanol:vinegar i1 (5
0:50) m solution, to which palladium, Yakol 1 oomp and ammonium formate 500 mp were dissolved.
added. The mixture was stirred at a high temperature for 10 minutes, then the palladium/charfur was separated, and the solvent was removed in 8 distillations under reduced pressure. The residue was treated with 50-trifluorotetraacetic acid containing 10% anisole for 1 hour, and then 15 mJ of methanol was added and left at 5°0 for 24 hours. The title compound was obtained by filtration of the resulting precipitate.

Example 4 (General formula (Ib-1): Preparation of Lexi 9-(2-(2-(carboxymethylaminocarl)ethyl)carbonyloxyethyl)purine) (A) - Obtained in Example 1 Glycine benzyl ester 4.951F (5G!111101.) suspended in 20 mA of 9-(2-hydroxyethyl)-hydrate methylene chloride and dissolved in 50 l of anhydrous salt 1 himethylene was added.The resulting reaction mixture The mixture was stirred at room temperature for 6 hours, filtered through F5, the PI was washed with saturated sodium bicarbonate solution 5Q+++4 and saturated brine, dried over sulfuric acid, and then evaporated under reduced pressure. The residue was purified by column chromatography on silica gel using methanol acetate (1:1) as a bathing solution. Further crystallization from ethanol yields 6-hydroxy-9-(2-(2-(benzyloxy)carbonylmethylaminocarbonyl)ethyl)carbonyloxy)ethylpurine 2.59C, which is a white powder crystal. Yield: 52% , m
p: 158-160°O).

Next, the characteristic values of the obtained compound are shown.

Elemental analysis value 202^lN506 Calculated value (support): o s6.2o u 4.91 N
16.59 actual value (4): o 56.51 H4,
75N 15.98MMR ((ODO/,
+1M9O-(L'), internal standard: Tus)2
．． 66J 4H (aiqlet) 0O-OH2-0%
-005,9Sa 2M(siqlst) MH-OH
,-004,40J 4H(broad single
t) N-0H2-OH2-05,25J 2M (i
■1et) 0H2-06)1゜' 7.26J 5H
(singlet) 06H57, 95 and 8. I
OJ 211 (singleta) purine hydrogen atom 1
0 J 2H (broad band) MH and 220 of the compound obtained in (A) of Example 4 above
mp (1,5 mmol 1a) was dissolved in methanol, and with stirring, 86 ammonium formate (1,6 mmol
e) and 100 rng of palladium/charcoal were added. The resulting reaction mixture was further stirred at 40°O for 1 hour, and 40 rng of ammonium formate was added.
Stirring was continued for another hour. TLO (ethyl acetate:
The older brother of the debenzylation reaction with methanol (1!1))
After checking, turn off the reaction mixture (celi)
te) and the R solution was removed. The obtained crude product was treated with ethanol, and the precipitate obtained by filtration was crystallized from a mixture of two bottles of methanol (90:10) to obtain the title compound (Ib-1197) as white crystals.
Yield +55%! aP i 186-190oO).

Next, the characteristic values of the obtained compound are determined.

Elemental analysis value 13H15N5°6 Calculated value)j O46,29H4,45N 20.75
Illusion value (%) +, 046j9 Ii4.25 N20.
97MMRX Pecth ((OD, QD + TEA
) Inside 49m1: TMS) 2.61SJ 411
(BinglLet) 00-Q+(2-OH2-0
05,98J 2N((siqlet) MH-OR,
-004,65J 4H(mlilplst)N-O
H2-OH2-08,2 and 9.25δ211 (s
inlets) Pooled hydrogen atom Example 5 of 6-hydroxy-9-(2-(2-(1-carboxy)ethylaminocarlyloxyethyl)carnyloxyethylpurine represented by (in the formula, PL is the same as above) Production] (4) 9-(2-
5.2 (hydroxyethyl)-6-hypotroxypropyl (hydroxyethyl)-6-hydroxypropanol (hydroxyethyl)-6-hydroxypropanol (1 ml) was added to 50 ml of dissolved methylene chloride. The resulting reaction mixture was incubated at room temperature for 6
After stirring for a period of time, it was treated in the same manner as in Example 4 (m) and crystallized from water to give white crystals of 6-hydroxy-9-(2-(2-(2-(1-benzyloxycarbonylethylamine)). carbonyl)-ethyl)carnyloxy)ethylpurine 2.6 g (yield: 57%, 1p;
140-142°O).

Next, the characteristic values of the observed compounds are shown.

Elemental analysis value "glH23'106 Calculated value (6): o57.14 N5.21 Iris 15
．． 87 Yomi value (6): 057.01 H5,01 irises 15.69 Seki spec F ((GDO/, 10 rMBo
) Medium, internal standard: Cabinet) 1.45δ51! (loubl
ei) OB, -am2.65δ48(I1・t
) co-C2-01112-C04,45δ11(
(quart@t) GH-1゜4.58δ4H(b
rod ainglet) N-(!H2-el12
-05.236211 (Yamabe/t) child, -06) 1゜7.25J5M (@Kyo1/t) 06H57,95
and 8.08δ2kl (siglets) fi
) N Mizu Abuko 10~11δ211 (broad si
gnal) 匪 and 0) １儂) Said *! 1,115 (226 m9 (0,511 m1o) of
) was dissolved in 20 ml of methanol and debenzylated in the same manner as in Example 4 (B). The title compound 103 was recrystallized from ethanol as a white crystalline solid.
(Yield: 57 times, P: 172-178°0).

Next, the characteristic values of the obtained compound are shown.

Elemental analysis value “14”lヮN506 Calculated value (%): 047.86 H4,88N21.
50 actual value (a): 048.18 H4,70I2
1.60mx vector) le ((GD30D + 'ryA
) medium, internal standard 1) 1.45δ5H (double
) OH, -〇)12.60δ4M(singlst
) oo-au, -tx2-o.

4.40δI H(qu&get) OH-01(3
4,65δ4K (broad ainglet)
l! −(14, −part 2−08.10δ 2H(sin
gists) Purine hydrogen atom Example 6 [General formula (Ik+-3): (wherein, !+1 is the same as above) M(x-
(Production of (5-(2-(6-hydroxyethyl-9)-ethoxy-carnyl)-propionyl)lysine] (AM
3.70 g (10 m mole) of ``-carbobenzyloxy-!-lysine-benzyl ester and 0.25 mz of triethylamine were dissolved in 50 m/11F of anhydrous methylene chloride to obtain a hydroxyethyl salt represented by the general formula (Vl-2). Added to suspension in methylene chloride.

The resulting reaction mixture was stirred at room temperature for 4 hours and then filtered. The p solution was distilled off under reduced pressure, the residue was treated with ethanol to form a white precipitate, and further crystallized from ethanol:water (9010) to t) M"-fikM>')l
Iit*S-Na-(5-(2-(6-hydroxypurinyl-9)ethoxycarbonylumprobionyl)lysine benzyl ester 3.2B.

(Yield: 52%, l1lp: 134-136°C) was obtained.

Next, the characteristic values of the obtained compound are shown.

Elemental analysis value "32" 36 N608 Calculated value (%) + 060.75 I5.69 I1
3.29 actual value (91;) EO60,55115,75
I13.20m spectrum ((opoz, + rsi
internal standard: IMS) 1-1.6δ6H
(mtltipl@t) C11(2-aH□-(!
) I22.6δ4H (slnglst) 0O-OH
2-GH, -C03,1δ2H (tripl@t)
QliE-11H4,4δ 511 (broad
ainglet) N-0H2-C4H6-0+CH
-COOk5.03 and 5.13δ41 (dou
bl@ ainglet) OHg-06H57,3δ
10H (s boundary 1@t) a6H57,95δ 2H
(siql@t) Purine hydrogen atom 9-11δ3H (
broad sign ml) MH,01 (@) The compound found in (4) of Example 6 was added to (to) of Example 4.
) was debenzylated in a similar manner to yield composition 403 of the title compound. Further, by crystallizing this from water, 545 mp of the title compound was obtained in the form of a white, hygroscopic powder.
(Yield: 76%, cape unknown) was obtained.

Next, the characteristic values of the obtained compound are shown.

MMR spectrum (in (Ili&30+D20), internal standard: 'IMS) 1.8δ6H (multipl@t)
0H2-GH2-OH22,6δ4H(mltip
lst) 00-(m, -0H2-003,2δ21
! (trlpl*t) OH2-MM23.6δ
1n (tripl@t) 0H-OH24,45
δ4H(siql@t) O-GH2-OH2-I8
．． 05δ2■ (8 boundaries xet) Purine hydrogen atom Example 7 (To-4) (In the formula, Pl is the same as above)
Production of 5-(2-(6-hydroxyethyl-9)-ethoxy7carbonyl)-propionyl)-M'-(4-carboxy-4-aminobutyl)guanidine] Lumamide 5
! lll&: N'-Z-/-arginine 33Dmp (1,07m mole) and dimethylformamide 4m/triethylamine 0,2m
/ was added to the mixture under stirring. After the obtained anti-element mixture was stirred for 5 hours, the solvent was removed and the resulting product was further purified using a gel furnace. The eluate was first freeze-dried, then dissolved in 50 ml of methanol, to which were added 100 ml of ammonium formate and 50 ml of palladium/charcoal, and the mixture was stirred for 3 hours at 40-45 ml. The resulting reaction mixture was filtered through a CELAI) and the solvent was saturated with He/. The resulting precipitate was then filtered, washed with isopropyl alcohol and dried under vacuum in the presence of phosphorus pentoxide to give 195 mg of the title compound (disintegration degree: over 200 oO ) was obtained.

Next, the characteristic values of the obtained compound are shown.

mx vector (in (IkIBO+DgO), internal standard: 'na) 1.78δ4H (mltiplat)
-0H2-11H2-2,55δ4H (single)
Co-0H2-OH [003,35δ2H(tr
iplet) CH2-MH-3,55δ1H(t
riplet) 0H-(!H24,40δ4H(
singlet) N-0H2-OH, -08,05
and 8.06δ2H (singl male) purine hydrogen atom Example 8 [General formula (Ib-5): N"-(1-
(Production of 2-(6-hydroxypurinyl-9)ethoxy-carbonyl)-propionyl)-tyrosine 181 mg (1 m !11016) of l-tyrosine was dissolved in 10 ml of deionized water with 400 ml of sodium bicarbonate. In addition, 540 yag of the chloride salt obtained in Example 2 was added thereto. However, the above reaction is 10°
I did it below C.

Once the reaction was complete (approximately 30 minutes), the pH was adjusted to 7 with 5% hydrochloric acid and lyophilized. The lyophilized product was then poured into hot anhydrous methanol, cooled and centrifuged, and the solvent was distilled off from the resulting supernatant under vacuum to crystallize white fine particles. The properties of this product matched those of the compound represented by general formula (Ib-5).

Next, its characteristic values are shown.

IMR spectrum (in m, internal standard: Thgs)2.
5δ411 (slglst) 0O-C14,-C
H2-CO3,05δ:2H(double) O
H, -CH3,98δ111 (ultipl@t)
0H-(Ji24.45611 (slglst)
)l-(!H2-cli2-.

6.6-7.0δ4H (string, BB') -06H4-7
, 9 and 8.0δ2H (-boundary 1・ts) Purine hydrogen atom 9δ1H (double) 11111.5δ2
M (broad band) CiA Example 9 6-hydroxy-9-(6-(2-(cartoxyethyl)aminocarbamoyloxy)hexyl)purine (1') represented by (in the formula, Pl is the same as above) I[i')
(4) 2.29 (9 mmole) of 9-(6-hydroxyhexyl)-hypoxanthine represented by formula (Itl-6) was suspended in anhydrous toluene 50+B.
5 mt of 0% phosgene solution (in toluene) was added under stirring. The resulting reaction mixture was stirred at room temperature for 3 hours and then placed under a nitrogen atmosphere to remove excess phosgene.
1.) was added to the solution of 30 m/&: methylene chloride. After stirring for 1 hour, the precipitate was filtered off, and the filtrate was distilled off under reduced pressure. The residue was dissolved in ethanol, filtered and crystallized from water to give a white crystalline powder.

(Yield: 7%, mp: 167-168°0) Next, the characteristic values of the observed compound are shown.

Elemental analysis value "u Hg? '505 Calculated value (6): o59.H6H6,12H15,87 Measured value (1): o59.60 lI6.02 N15
．． 65 Karasuta Spec Furu 0-hai Genchu, Internal Standard: TMS
) 1.35δ8M (corp ■l@t) c-(ca2)
, -cl, 48δ3H (double) CH3-
CH4,00δ4H (2 triplets) cH2
−○ and CH2−N4+45δ1■ (quartet
) an-an.

5.25δ2H (Company ■1・t) 15 to CH2-06
7.30δ5M (-boundary l@t) C! 6m (57,8
0 and 8.30δ2H (-¥1sts) Pudding water #Kuko 9-11δ211 (brom slP&l)
□H, MH@) Compound 440m obtained in the above (A)
g (1 mmol.) was debenzylated in the same manner as in Example 4 -) to obtain 318 mp of the title compound as a white powder (yield: 90%, Otori) p l > 22
000).

Next, the characteristic values of the obtained compound are shown.

Elemental analysis values: O engineering, 1121M, O.

Calculated value (2)) 2051.28 H5,9B) 1
19.94 collection 11 value (%): osl, oo N6.
02 N20.51M spectrum ((rMBo+D2
0), internal standard: 'nas) 1.33δ8H (
brom*iql*t) 0-(aH2), -01,4
5δAI (doublest) OH, -(N6.9
4δ2H (triplet) CIHz-N4.12
δ2H (triplet) (IH2-04,40δ
11 (quartst) GH-CH37,95 and 8.12δ2H (aingl·ta) Purine hydrogen atom Example 10 6- and droxy-9-(5-(2- Preparation of (carboxyethyl)amino-carbamoyloxy)pentyl)purine] (4) Same as Example 9 (A) using 9-(5-hydroxypentyl)-hyboxanthin represented by formula (11-5) By the method of
Oxy)pentyl)purine (mpq1B4°0) was obtained.

Next, the characteristic values of the obtained compound are shown.

Elemental analysis value = 021 H! ! 5'5 o5 calculated value (5): 057.66 N5.72 F16.01
Actual value 61G): o57.51 N5.62 N1
6.20 side enemy spectrum (rkMJ medium, internal standard: Ai)
1.36δ611 (broadsiglst) o-
(an2)3-cl, 4563H (loubl@t)
character, -(H4,03δ4H(2triplsts)
0M2-0 and 0H2N4.43δ1H (quar
t@t) >-open.

5.23δ2H(siql*t) 0H2-0615
7,3δ5H (ainglet) a, a.

7.80 and 8.28δ2H (ainglet)
Purine hydrogen atom 9-11δ 2H (brom miga
al) OH, Ni) Using the compound obtained in (4) above, a compound (decomposition temperature: 200 o'O) was obtained in the same manner as in (II) of Example 9-. .

Next, the characteristic values of the obtained compound are shown.

Elemental analysis value 14 N19 N5 o5 calculated value Trowel)+
049.85 N5.65 N20.87 fresh measurement value (
%) + a49. so N5.82 N2[1,4
5M spectrum (IMBO, D20 medium, internal standard: Seki)
1.55δ6H (broad single@t)
C-(0112)3-01.45δ3H(dOubls
t) OH, -0H3,95δ2H (triplet
) (!H2-M4.1262H (triplet)
CH2-04, 40δI H (quartet)
0H-OH37,95 and 8.10δ2H (sin
glets) Purine Hydrogen Atom The compounds of the present invention have specific immunomodulatory effects that are superior to drugs such as methitubrinol (m@thi@opri11o1), which are already widely used in the field of clinical medicine.

Figure 1 shows Hodgkin's disease.
Comparison with Compound (Ib-4) when the enlarged and enlarged lymphocytes of patient e) were rejuvenated by phytohemagglutinin (hereinafter referred to as PHA) at least 17 months after discontinuing treatment. 1 is a graph showing the general tendency of the effect of methytubulinol on the development of lymphocytes. The horizontal axis of the graph shows the concentration (r/wm z ) of the compound-*(Ib-4) and the comparative substance metitubulinol added to the medium, and the vertical axis shows the concentration (r/wm z ) of the compound-*(Ib-4) added to the medium, and the vertical axis shows the concentration (r/wm z ) of the compound-*(Ib-4) added to the medium. Labeled thymidine (
Hereinafter referred to as ~-thymidine), the amount of incorporation is opm (
ount per adnuts).

In addition, PHA is concentrated, 01ioj/thought! It was used in P.H.
The rejuvenation reaction of lymphocytes caused by A is lower than the normal range in lymphocytes of Hodgkin's disease patients.

A compound (Ib) that affects the development of lymphocytes by PHA
-4) and methytubulinol in 1n mice were investigated by culturing lymphocytes that had been made into infants with PHA in the presence of various concentrations of these compounds. The concentration used was methitubrinol: 100.
200.400rlre i compound (Ib-4):
0.1.0.5.115γ/mt.

As is clear from Figure 1, compound (Ib-4) has a 0.1
At a concentration of r/l, methitubrinol already enhances the blastogenesis of lymphocytes induced by PHA, whereas methitubrinol has a concentration of 2
Even at 00 to 400 r/ml, it shows only a lower enhancing effect than compound (Ib-4).

More importantly, the concentration of 0.1 to Ii/m/ at which Compound (Ib-4) exhibits an enhancing effect is a physiological concentration and can be applied in vivo. On the other hand, the concentration at which methitubrinol exhibits the maximum effect is too high for use in living organisms.

Therefore, the results of this experiment indicate that compound (Ib-4), one of the compounds of the present invention, can be used for more effective treatment of tumor tumors than methytubrinol.

FIG. 2 is a graph showing the effect of a compound (I b-4'') on the oxidative metabolic activity of the membrane of imaginalized polymorphonuclear lymphocytes.

That is, polymorphonuclear leukocytes (2°5 cells x 10'/m4) were mixed with Am 50 in a phosphate buffer (hereinafter referred to as PI3).
After adding 0.125 m/m of superoxide dismitter, 7-erythytochrome 0 (1.5 mp/m) was added in the presence or absence of SOD (hereinafter referred to as
/) at 5700 for 15 minutes. Then, the amount of reduced cytochrome O in the supernatant from which cells have been removed (O.11fr..) is measured by absorbance at ssonm, and the amount of reduced cytochrome O is determined by the absorbance at ssonm according to the standard formula. - The amount released was calculated. The horizontal axis of the graph is the concentration of compound (Ikl-4) [
(r/+mr), the vertical axis indicates the absorbance at 550 nm.

From the results of statistically processing the data obtained above using the T test, it was found that compound (lb-4) has a statistically sufficient effect of promoting superoxide production at a concentration of 1ΔD/mt. It became clear.

When the number of guests becomes higher than that, the amount of 02- produced decreases gradually, probably due to the complex mechanism of metabolic control. The compound (To-4), which shows more effective oligophagocytosis by increasing the amount of superoxide anion produced, has a concentration of 0.001 to 40μ,/I
#b cells of 8- to 10-week-old 1-mu LB10 mice were rejuvenated by PH-mu or lipo-polysaooharl saccharide (hereinafter referred to as LPf9) at one time in 1/2/2015, and 3H-thymidine incorporation was performed. It also has the effect of inhibiting Compound (Ib
-4) The inhibitory effect on stem cell proliferation in iKL VitrQ is dependent on use (dome-4@p@nd・nt)
This effect is probably due to suppressor cellular mechanisms.

Next, a compound (I
Table 2 shows the results of investigating the effect of compound (Ib-4) on suppressor cells. That is, the lymphocytes were incubated with compound (Ib-4) at various concentrations for 48 hours and then washed.

The cells were then incubated for 5 days with autologous lymphocytes transformed with concanavalin A (hereinafter referred to as oonA) or with xenogeneic lymphocytes treated with mitomycin for 5 days. The concentration of the compound (tb-4) used was 0.1-1
．． The concentration was 5N/ml, and the growth inhibition effect was approximately 60% compared to control. Furthermore, T of the compound of the present invention
+ Activity of IJ member ball 1-roset
The effect on the ability to form tes) was determined by Pacino et al.
mo #coil) (Boll, 工SO, 81
*ratsr.

11111.60, 4.297-501 (1981
)).

Briefly, lymphocytes were incubated for 24 hours at 4°O and then incubated for 90 minutes in the presence or absence of various concentrations of test compound before being tricked and analyzed. The result is the percentage of cells that formed a-zet (6)
It was expressed as Compound (Ik+-2) was 35-50% at 1-6p9/-, and at higher concentrations the percentage of active rosettes decreased.

Furthermore, the compound of the present invention was tested in 121 vivo ((! 57
When administered to Bll/6 mice at 0.1 to 5 cm), it has the effect of increasing anti-sheep red blood cell antibody and anti-TNI (linitrophenyl)-LPS antibody.

According to the trypan blue exclusion test, the compounds of the present invention:
60-12O in culture of human mononuclear peripheral blood cells
It was found that there was no cytotoxicity (survival rate of 98% or more) even at a very high concentration of N9/mj.

[Brief explanation of drawings]

Figure 1 shows that the compound (Ib-4) and the comparative compound methytubulinol were found in some patients with Nezkin's disease when the bulbs were rejuvenated by PM people after at least 17 months of treatment. Figure 2 is a graph showing the effect of a compound (Ik+-4) on the oxidative metabolic activity of the membrane of young lymphocytes. be. Special N 11 people Thio Pharma Kohoshi Shien Figure 2+ -u'?

Claims (1)

[Claims] 1 General formula (I): ? --(%)nOR' (1) is a hydrogen atom or a town) is a do W now sibrin group,
Otori is an integer from 1 to 6, a nucleotide, a hydrogen atom, a methyl group, P
-hydroxybenzyl group or aminopentyl group, R3
is a hydrogen atom or an oligopeptide residue having 2 to 6 amino acids), generally an integer of ~6, R2 is the same as above, - is a hydrogen atom or an oligopeptide residue having 2 to 6 amino acids; Residues represented by (representing peptide residues), residues represented by general formulas (same), especially n
wherein p is an integer of 2 to 6, R2 and R3 are the same as above) or a compound represented by the general formula: or a compound represented by the formula (Ib-4): R. 2. The compound according to claim 1, wherein the a-amino acid is an optically active 1-amino acid, a d-amino acid, or a racemate. 6. The compound according to claim 1, wherein the amino acid is a natural amino acid. 4 The compound where n is 2, R is a hydrogen atom, and R1 is as described in . 5 The above n is 2, R is a hydrogen atom, and R1 is -ao-(~,
The compound according to claim 1, which is -0〇-order -Q%, -000H. 6. The compound according to item 1, wherein n is 2, R is a hydrogen atom, and R1 is a range. 7. The compound according to claim 1, wherein n is 2, R is a hydrogen atom, and R1 is H. 8 The compound according to claim 1, wherein n is 2, R is a hydrogen atom, and R1 is H. 9 The above n is 6, R is a hydrogen atom, and R1 is a compound as described in 10. The above n is 5, R is a hydrogen atom, and R1 is a compound as described in 11. General formula (1): % formula % () is hydrogen a hydroxypurine residue (n is an integer from 1 to 6);
n' -M -on -000H(1)2 (In the formula, R8 is a-amic) acid, hydrogen atom, methyl group, ?
-hydroxybenzyl group or aminopentyl group, RB
is an oligopeptide residue having 2 to 6 amino acids in which the amino group usually used in peptide synthesis or the terminal amine 7 group is protected. a-amino acid oligopeptide by activating the carmexyl group using known reagents such as M,N'-carbonyldiimidazole, dicarbodiimide, and similar compounds thereof to obtain the general formula (5): General formula (I) consisting of preparing an intermediate represented by (wherein PsnsR2 and R5 are the same as above) and then removing the amine protecting group by a known method: P-(OH8 -o-R' (1) (wherein P and n are the same as above, R1 is a hydrogen atom or an oligopeptide residue having 2 to 6 amino acids) ) Preparation of the compound represented by the formula (II) 2p-(OH2)n-〇H (1) 1 is a hydrogen atom or MEI2) Hydroxypurine residue represented by the formula (II), n is 1 to 6 is an integer) and an activated dicarboxylic acid derivative to react with the general formula (V): (wherein P and n are the same as above, and m is an integer from 2 to 6. An intermediate represented by the above is prepared, and then this intermediate is combined with α-adenyl or 171/acid, water atom, methyl group, hydroxybenzyl group, or aryonopentyl A, R
It consists of reacting an oligopeptide with a protected carboxyl group represented by 'C', which is an oligopeptide residue having 1 or 2 to 6 amino acids, by selectively using a known activator. A method for producing a compound represented by the general formula (I): p (OH2)nOR' (1) (wherein P and n are the same as above, R1 is general and R' is the same as above). 16 General formula (M): hydroxypurine residue represented by p-(OH2)n-OH(1) OH (represents a hydrogen atom or town), n is an integer from 1 to 6) 6-
Hydroxypurine and phosgene are reacted, and then α-
Azoic acid or a-amino acid, hydrogen atom, methyl group, hydroxybenzyl group or aminopentyl group, R6 is a carboxyl group protecting group or an oligomer with a terminal carboxyl group having 2 to 6 amino acids protected A method for producing a compound represented by the general formula (1) (which is a peptide residue): % formula % (1) (wherein ! and n are the same as above, and R1 is the same general formula). 14. General formula (1): % formula % (1) represents a hydrogen atom or town) is a droxypurine group, n is an integer from 1 to 6, an amino acid, a hydrogen atom,
(where m is an integer of 2 to 6, - is The same as above, R4 represents an oligopeptide residue having 2 to 6 amino acids), and the rest - is the same as above), especially when - is 1. General formula: a residue represented by the number, - and R3 are the same as above) or a residue represented by the general formula: psR" and R4 are the same as above))) or a compound represented by the formula %) ) A pharmaceutical composition having an immunoarticulatory effect, comprising at least one compound having an immunoarticulatory effect.