JPS60116700A - Kyotorphin derivative - Google Patents

Abstract

NEW MATERIAL:A compound of formula I {R1 is H, lower alkyl, acyl; R2 is OH, lower alkoxy, acyloxy, halogen; R3 is H, NO2; R4 is OH, lower alkoxy; aralkyloxy, acyloxy-lower-alkoxy, glycine residue, thyrosine residue, thyrosine amide residue, phenylalanine residue, thyrosine ester residue, formula II [R5, R6 are H, (hydroxy)-aralkyl, (hydroxyl, lower-alkoxyl, lower alkylthio-) lower alkyl]}. EXAMPLE:The compound of formula III. USE:Analgesic. PREPARATION:The condensation reaction of an arginine derivative of formula IV(R7, R8 are protecting groups) with H-R4 such as an amine, aminoacid or alcohol gives a comopound of formula V followed by deprotection of the group R7. Then, the product is subjected to condensation with a thyrosine derivative of formula VI (Ra is protecting group), followed by deprotection of R9 and R8 to give the compound of formula I where R1 is H.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel kyotorphine fatty conductor represented by general formula (1).

In the formula, R, means H9 lower alkyl or acyl, &
means hydroxyl, lower alkoxyl, acyloxyl or halogen, R3 means H or nitro, R4 means hydroxyl, lower alkoxyl, aralkyloxyl, acyloxy lower alkoxyl, glycine residue, tyrosine residue, tyrosine amide residue, Phenylalani/R5 means a tyrosine ester residue, a tyrosine ester residue, or a group represented by the formula -N8 (R5 and R5 respectively represent H, aralkyl, and hydroxyaralkyl).

phenyl, lower alkyl or hydroxyl.

(lower alkyl or - means lower alkyl substituted with lower alkylthio).

Acyl groups in R+ include those derived from aliphatic, aromatic or heterocyclic carboxylic acids, such as acetyl, propionyl, butyryl, pantoyl, benzoyl,
Examples include N-substituted dihydronicotinoyl. & In addition to H, lower alkyl, phenyl, aralkyl, hydroxyaralkyl, and alkoxyl.

Examples include lower alkyl substituted with argylthio or hydroxyl.

Takagi et al., one of the inventors of the present invention, isolated the endogenous analgesic substance kyotorphin L-Tyr-L-Arg from bovine brain (
H. Takagi at al., Nature.
282.410 (197o)) 〇Kyotorphin is an opioid in mice, a peptide, methionine ◆ Enkephalin 44? I
It has analgesic effect.

K.-Dolphin is antagonized by opioid receptors. The analgesic efficacy of Kyotorhuin is thought to be expressed through the release of enkephalin, and it is expected to be a new type of analgesic.

The present inventors aimed to obtain a compound that is economically advantageous, stable as a compound, and has a sufficient analgesic effect.As a result of extensive studies, the present inventors found that some kyotorufine derivatives have a stronger analgesic effect than the original compound. We have discovered a compound that has this effect.

The dolphin derivative (I) of the present invention can be produced by various methods, for example, as follows.

That is, first, an arginine derivative represented by the general formula (in the formula, R1 represents a protecting group for an amino group, and R11 represents a protective group for a guanidino group) and an amine, amino acid, or alcohol represented by the general formula %(). After removing the protective group R7, the compound represented by the general formula produced by condensing is condensed with the tyrosine derivative represented by the general formula (R9 represents a protecting group for an amino group) to form a compound represented by the general formula. The compound of general formula (1) (where m means hydrogen) is obtained by producing and then removing the protecting group RQ and, in some cases, the protecting group contained in the compound by conventional deprotecting means. Can be done.

In order to obtain a compound in which R1 is an alkyl group or an acyl group, appropriate raw materials may be selected and the reaction may be carried out using known means.

In the above general formula, the protecting group for the amine 7 group represented by R7 or R9 is 2, for example, benzyloxycarbonyl,
p-Nitrobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-phenylazobenzyloxycarbonyl + p-(p-methoxyphenylazo)-
benzyloxy, carbonyl, p-chlorobenzyloxycarbonyl, p-bromobenzyloxycarbonyl,
p-tolyloxycarbonyl, α-naphthylmethoxycarbonyl, p-dodecyloxybenzyloxycarbonyl, tertiary butyloxycarbonyl, isobornyloxycarbonyl, adamantyloxycarbonyl, diphenylphosphinyl, diphenylphosphine/thioyl group, etc. In the above general formula (V), when R2 is a hydroxyl group, it may be protected with an acetyl group, benzyl group, benzyloxycarbonyl group, tert-butyl group, etc., but it is not always necessary to protect it. .

The protecting group ratio of the 7 guanidi groups in the above general formula (It) is 9, for example, a nitro group, a tosyl group, a p-methoxybenzenesulfonyl group, a mesitylene-2-sulfonyl group,
Examples include a benzyloxycarbonyl group, an isobornyloxycarbonyl group, and an adamantyloxycarbonyl group.

The acid amide production reaction in the above general formula can be performed using conventional peptide bonding methods, such as the azide method, mixed acid anhydride method, dicyclohexylcarbodiimide method, and active ester method. This can be easily carried out by a method using Woodward's reagent K, a carbodiimidazole method, or the like.

These reactions are generally carried out in a known solvent that can be used in peptide condensation reactions, such as dimethylpolamide, chloroform, ethyl acetate, dioxane, dichloroethane, tetrahydrofuran, or a mixed solvent thereof. The reaction temperature for this reaction is not particularly limited and can be appropriately selected within a wide range, but it is generally preferable to carry out the reaction at a temperature of about -80 to 80°C.

Esterification and alkylation in the above general formula.

Acylation etc. can be easily carried out using known means.

Examples of deprotecting methods for removing all or part of the protecting group in the above general formula include catalytic reduction using palladium black, palladium carbon, platinum, etc. as a catalyst, trifluoroacetic acid method, hydrogen bromide method, hydrogen chloride method, etc. method, hydrogen fluoride method, methanesulfonic acid method, trifluoromethanesulfonic acid method, etc. These methods generally range from 0 to 40
It is carried out at temperatures around ℃. In addition, in the method using an acid, it is effective to add a cation scavenger such as anisole or thioanisole.

The compound of formula (1) thus obtained can be processed by means commonly employed to separate peptides, such as extraction, reprecipitation,
Silica gel chromatography.

It is easily isolated and purified by ion exchange chromatography, countercurrent distribution, gel chromatography, affinity chromatography, etc.

The compounds of the present invention can easily form acid addition salts with pharmaceutically acceptable acids, and such salts are also included in the compounds of the present invention. Examples of acids that can be used include hydrogen halides such as hydrogen chloride and hydrogen bromide, sulfuric acid, nitric acid, oxalic acid, maleic acid, citric acid, tartaric acid, and vinegar.
Examples include organic acids such as te p-toluenesulfonic acid.

Incidentally, the compounds of the present invention include both optical isomers and racemates.

Production examples of the compounds of the present invention are listed below as Examples. In addition, the following abbreviations are used in the examples. L-type amino acids were used unless otherwise specified.

Arg: arginine Tyr: thytetracine Phe: phenylalanine' phe (p-F): parafluorophenylalanine G,ly nigericin z (oMJ: paramethoxybenzyloxycarbonyl Bzl: benzyl OBzl: benzyl ester OTCP: 2,4.5 - ) Lichlorophenyl Esder DMF Nidimethylformamide TFA :) Lifluoroacetic acid DCHA Nidicyclohexylamine HONB : N-Hydroxy-5-norbornene-2
．． 8-dicarboximide or Rj value is silica gel [Merck, Kieselgel 6]
Thin layer chromatography (TLC) on
) using the following mixed solvent.

Rf bichloroform-methanol-water (S:8:1
) Mixture liquid Rf'3-n-but/-ru: #ethyl acid: pyridine: water (i:i:i:i) Example I H-Tyr-D-Arg-NH(OH2)a
06) Production of Is (compound l)■Z(0M6)-o-*rg(NO2)-NH(oH
l) acsH5z(ova)-o-Arg(Noz)-
oHl, 909 was dissolved in DMF5-. Under cooling with water, 0.71 t of triethylamine/-isobutyl chloroformate □, 72- was added and stirred for 15 minutes. Next, ToN
-(GHz)3-csH5o,s5- was added, and the mixture was reacted for 18 hours while gradually returning to room temperature. The solvent was distilled off, ethyl acetate was added to the residual aroma, and 5% citric f! i? s 5% sodium bicarbonate solution.

After washing with saturated brine and drying over IJ sulfate, ethyl acetate was distilled off. Crystallize by adding diethyl ether,
Recrystallized from ethyl acetate-diethyl ether. Collection Wz,t
ag, melting point 52-62°C.

Rfm O, 69, (α)24 + 2. +°(c-1
, Q, ethanol).

■Z(OMe)-Tyr(Bzl)-n-Arg(
No2)-in(CHz)ac6HsZ(OMe)-D
-Arg(NO2)-NH(OHz)a(jaHs 1
．． 009 as anisole-TF A (1,1d-2,
The *2 (0λ(13) group) was removed by a conventional treatment with 8 m/
A DMF solution 2- of r(Bzl)-0T Ci Pl, 299 was added and reacted at room temperature for 18 hours. Distill the solvent,
Add diethyl ether and water to the residual aroma, and filter out the white crystals that precipitate.

It was recrystallized from methanol. Collection ff1i, oo9. Melting point 151-154℃e RflO074e (α),;'
Tengg, e.

(c-1,0,THF).

■H-ryr-n-Arg-NH(cH2)a C6H
6z(oMe)-Tyr(Bzt)-n-*rg(N
Dissolve O2)-NH(ri)b) aosHs170■ in methanol-acetic acid (20d-1+/) and Pd-black 20
Catalytic reduction was carried out using 0. Catalyst filtered off. The filtrate was concentrated and powdered with n-butanol-diethyl ether. This was dissolved in a small amount of 0.2N acetic acid and subjected to Sephadex G-10 column chromatography using the same solvent as the eluent. Take a sample every 8-minutes and measure the absorbance at 275 nm. Chive numbers 19 to 24 were collected and concentrated, and the residual aroma was powdered with n-butanol-diethyl ether.

80 ■.

Rfz O,60+ (α) +28.8° (C-t
, O, methanol).

Elemental analysis a24Hs4NaOa・2AcOH4H20
Calculated value as 058.06. )l 7.40. N
14.51 Experimental value C57,88,)47.61.
N 1421 Example 2 H-Tyr-D-Arg-0(
GHz)zGaHs (Compound 11) Production ■Z(OMe)-D-Arg(NO2)-0 (0%)
206H5Z(OMe)-D-Arg(M)-0H1,
5s 9 in methanol-water (8Q d -3tnl)
It was dissolved in a mixed solution, and a 20% AS2CO3 solution was added thereto to neutralize it. The mixture was concentrated under reduced pressure, and approximately 18 mm of DMF was added in two portions to the colander, followed by concentration under reduced pressure to completely remove methanol. Add to this Br-(C)b)z=C6&o,s8g(7
) DMF (20gLt) solution was added and reacted for 2 days. The reaction solution was concentrated under reduced pressure, water was added, and the mixture was extracted with ethyl acetate. Wash with water and saturated saline. After drying with IJ sulfuric acid, it was concentrated to obtain an oily substance.
l 0.57゜■Z(OMe)-Tyr-n-Arg
(Nc)-0(C)b)2061sz(oMe)-n-
Arg(No2)-0(GHz)zlaHs 1.87
9 to anisole-TFA (1,51117-2,
2d). Solvent evaporation. Add n-hexane to the residual aroma and remove the supernatant. The residual oily substance was dried with potassium hydroxide granular soil. This was dissolved in DMF8-, and z(oMe)-Tyr-N was added to a solution to which 0.4-triethylamine was added.
HNH21.01g# 8.87 NHC//D M
F<1. a-) isoamyl nitrite 0.421111
j,) 1.3 gL of ethylamine! A more prepared solution of azide in DMF (6-) was added, and the mixture was reacted at 4°C for 48 hours. The solvent was distilled off, the residual aroma was dissolved in ethyl acetate, washed with 5% polyenic acid, 5% aqueous sodium bicarbonate, and saturated brine, dried over sodium sulfate, and then ethyl acetate was distilled off. N-hexane was added to the residual aroma to form a powder. Yield: 1.5 kg, melting point: 98-1 (11℃,
RtlO, 85.

[α] is also '+10.8° (c-i, o, methanol)'
.

■H-Tyr-D-Arg-0(OHz)z CsHs
Z(OMa)-Tyr-D-Arg(No2)-0(
C)I2)z 700J of CaHs (1) was dissolved in 25m of methanol, 2g/m of acetic acid and 850m9 of acetic acid were added, and the mixture was exposed to light for 24 hours.

Catalyst filtered off. Thereafter, the same treatment as in Example 1-■ was carried out to obtain the desired product. Yield 444”9 + Rfz O-78+
[α)”+ a a, oo (0-1,0, methanol).

Elemental analysis Oz s Ha I Ns Oa -0H3
C00H-1,5I20 and shite calculation value Cj 56.8
0. H7, 25, N 18.25 Experimental value G 56.
80. H7,04,N 18.28 Example a H-T
Preparation of yr-D-Arg-Phe-OH (compound 18) Oz(ova)-n-Arg(Noz)-phe-oB
ziz(OMe)-n-Arg(NO2)-0)(8,
809 in DMF7-) diethylamine 1,4
t+F, acid isoamyl 1.4- is added to make a mixed acid anhydride, and H-Phe-OBzl・Tos-OH 5,1
8g of triethylamine 1. ? D neutralized by wLt
Add MF solution and react for 18 hours. Solvent evaporation. Dissolve the residual scent in ethyl acetate and wash with 5% citric acid, 5% sodium bicarbonate solution, and saturated saline. After drying with IJum (sulfuric acid), ethyl acetate was distilled off, and diethyl ether was added to form a powder. Collection'IiL
+. osg, melting point 88-84°C, Rf+O. 6 8
, (α) H5- 7.8° (C-1.0, DME)
.

■Z(OMa)-Tyr(Bzl)-D-Arg(
NOz)-Phe-OBzlz(OMe)-n-Ar
g(NOz)-Phe-OBzl 1. 55 g of anisole-TFA (1.id-9,7nj
) for conventional processing. Add diethyl ether to make a powder.

Filter and dry. Dissolve this in DMFIO-, add 0.85m of triethylamine, Z(OMa)-Tyr(Bzl)
-OTCPl. Add 61g and react for 18 hours. The first solvent was distilled off, and diethyl ether was added to the residual aroma to make a powder. After washing with 5% citric acid and water, the powder was reprecipitated from DMF-diethyl ether. Number position 1. t5g, RfmO. 84, melting point 1
82~134°c, (α)25-1 1.0'(0-1
．． 0,DMF).

■H-Tyr-D-Arg-Phe-OH2 (OMa
)-Tyr(Bzl)-n-Arg(NO2)-Ph
Dissolve 50g of e-OBzll in methanol 25-
Vinegar i! 1112m1, add PD-black 1.09 and contact light for 24 hours. Thereafter, the same treatment as in Example 1-■ was carried out to obtain the desired product. Yield 5 6 01 to 9, Rfz0.5 2
, (α);'+5 0.4.

(0-1.0, methanol).

Elemental analysis +a48szNsos・CI(3C00H-
Calculated value as I20 C 55.50, H 6.81
, N 14.94 Experimental value G 55.60, H 6
．． 98, N 15.48 Example 4 H-Phe(p-
F) Preparation of -D-Arg-OH (Compound 19) Boa-Phe(p-F)-0H-DCHA 1.8
89 was dissolved in ethyl acetate 20-L and treated with 0.5N sulfuric acid 20-L. Wash the ethyl acetate layer with saturated brine.

After drying with sodium sulfate. Solvent evaporation. This formula is THFIO
-Dissolved in HON8 564'9t DCo 6 5
Add 0''9 and react for 8 hours. Insoluble matter was filtered off.

H-D-Arg-OH-NO17 2 8 11 to the filtrate
An aqueous solution 8 containing 0.48-triethylamine of 9 was added and reacted for 24 hours. Solvent evaporation. Add 8% acetic acid to the residual aroma and extract with n-butanol. After washing the n-butanol layer with water, the solvent was distilled off. Diethyl ether was added to the residual fragrance to form a powder. Acquisition M1.08ri.

Of these, 500119 in the presence of 0.5 anisole,
Regular processing using TFAX-. Diethyl ether was added to the reaction solution to form a powder, which was then filtered and dried. This was dissolved in 10 - of water and treated with Dowex 1 x 4 (ACO - about 1) to give an acetate salt. Thereafter, the desired product was obtained in the same manner as in Example 1-(2). Gai 227■, Rf2 o. 41 7 m
(α); 5+ a 2.4.

(C-0.4, methanol).

Elemental analysis CtsH2zNsOsF-GHaCOOH-
Calculated value as +HzO 0 49.99, H 6.6
6, N 17.15 experimental value C 49:48, H 6
．． 64, N 17.68 Example 5 Pantoil-Ty
H-Tyr-D-Arg-NH(C) obtained in Production Example 1 of r-p-Arg-NH(C)(2)306H5 (Compound 20)
I(2)3CaH5800■ was dissolved in methanol 6- and triethylamine was added to adjust the pH to about 9.

Vantolactone 888 .mu. was added and the reaction was allowed to proceed overnight at room temperature. Solvent evaporation. The residual aroma was dissolved in a small amount of 0.2N acetic acid, and the residue was purified by Se7Adex G-10 column chromatography in the same manner as in Example 1 to obtain the desired product. 248
A9. Rf20.82゜[α]: 5+ a s, oo
(C-0,4, methanol).

Elemental analysis G3oH4aNaos・0)LqCOOH4
1 (calculated value as 2o C58, 79, H7, 56, N
12.86 Experimental value C58,58,H7,46,N
12.31 Kyotorphin derivatives prepared in the same manner are shown in the table. In addition, the configuration based on two asymmetric carbon atoms in general formula (1) shall be expressed as L-D, L-L, and if it is necessary to indicate the configuration in the Rt part, it will be shown in the table. . Also, ratio table 1 in Table 1: (R1”'4(, R
2-0H, R3-H, steric configuration table 2: (R1-H,
R2-0H, R3 is A18 is NO2 Others are H9 configuration is L-D) Table 8: (Position is A19 is F Others are OH, R3-H+ configuration is A25 is L-L, Others are L-D type ) It has been confirmed that the kyotorphin derivative of the present invention exhibits strong analgesic effects in mice.

It is expected to be useful as a medicine.

Next, a test example on mice will be shown.

Test Example (10 mice (14-17 g, male) were used in each group. The test compound was dissolved in physiological saline, and a special injection needle (needle length: 5 cm) was injected into the cerebral cisterna of unanesthetized mice. The analgesic effect was obtained using a hypodermic needle (made into a J-shape).
Japanese Journal of Phar
Macology 16.287-294 (1966)
) was tested. All test compounds exhibited dose-dependent analgesic effects, and the 50% effective dose for these analgesic effects (
RDso) is as shown in the following table.

In addition, the LD50 of these compounds when administered intravenously to mice is 200 μ/lC9 or more, and when administered within this range, no abnormal behavior or respiratory depression was observed, and there was no guinea pig ileal constriction effect, and the compounds of the present invention had no effect on morphine. It is highly anticipated that it will be a safe and useful analgesic drug that does not have undesirable side effects such as constipation, respiratory depression, decreased blood pressure, and miosis. Also, as seen in the test results above.

The effective dose of the compound of the present invention is comparable to that of morphine, and the dose of 1 morphine is extremely small at 5 cm at a time and 15 cm per day, and the compound of the present invention has low toxicity. Considered suitable for use.

Claims (1)

[Claims] A compound represented by the general formula. However, RI means H1 lower alkyl or acyl, 几 means hydroxyl, lower alkoxyl, acyloxyl or halogen, and R3
means H or nitro, R4 is hydroxyl, lower alkoxyl, aralgyloxyl, acyloxy lower alkoxyl, glycine residue, tyrosine residue, tyrosine amyl' residue, phenylalanine residue, tyrosine ester residue or with the formula -N7h ((几) and 鈥擭 means respectively H, aralkyl, hydroxyaralkyl, phenyl, lower alkyl or lower alkyl substituted with hydroxyl, lower alkyl or lower alkylthio).