JPH0322398B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel tetrapeptide derivatives having analgesic activity. Hughes et al. [Nature, vol. 258,
577 (1975)], two types of pentapeptides called enkephalins, namely H-
Tyr-Gly-Gly-Phe-Met-OH and H-
Tyr−Gly−Gly−Phe−Leu−OH is separated,
Their chemical structures have been determined, and although these substances exhibit morphine-like effects when administered directly into the brain, they do not exhibit analgesic effects when administered intravenously. On the other hand, β-endorphin, which is considered to be derived from the carboxy terminus of β-lipotrophin, clearly shows analgesic effects even when administered intravenously, but this substance is a polypeptide consisting of 31 amino acid residues, and its organic chemical It is extremely difficult to manufacture by synthetic synthesis, and it is extremely difficult to supply large quantities as pharmaceuticals. The present inventors conducted various studies in order to obtain a compound that is economically advantageous, stable as a compound, and has sufficient analgesic effect even when administered intravenously or subcutaneously, and found that a specific compound of a tetrapeptide hydrazide derivative found that it is compatible with the above purpose,
As a result of further research based on this knowledge, the present invention was completed. That is, the present invention provides (1) general formula () [In the formula, R ₁ represents hydrogen. R ₂ represents the side chain of D-α-amino acid. R ₃ represents hydrogen or alkyl having 1 or 2 carbon atoms. R ₄ is hydrogen, a linear or branched alkyl having 1 to 8 carbon atoms substituted with a hydroxyl group, a hydroxyl group, an amino group,
It represents a linear or branched saturated aliphatic acyl having 1 to 8 carbon atoms which may be substituted with alkoxy having 1 to 4 carbon atoms or halogen. ]
Tetrapeptide derivatives represented by and pharmacologically acceptable acid addition salts thereof, and (2) general formula () [In the formula, R ₁ represents hydrogen. _R'2 represents a side chain of D-α-amino acid which may have a protecting group. R ₃ represents hydrogen or alkyl having 1 to 2 carbon atoms. R' ₄ is hydrogen, a linear or branched alkokyl having 1 to 8 carbon atoms substituted with a hydroxyl group, or a straight chain which may be substituted with a hydroxyl group, an amino group, an alkoxy having 1 to 4 carbon atoms, or a halogen. It represents a chain or branched saturated aliphatic acyl having 1 to 8 carbon atoms, which may have a protecting group. This is a method for producing a tetrapeptide derivative represented by the general formula (), which comprises subjecting the compound represented by the formula () to a protecting group elimination reaction. Amino acids and peptides as used herein are those commonly used in the art or IUPAC
- The abbreviations adopted by the IUB Nomenclature Committee are used, for example, the following abbreviations: Ala: Alanine Gly: Glycine Leu: Leucine Phe: Phenylalanine Me-Phe: N-methyl-phenylalanine Tyr: Tyrosine In addition, commonly used compounds are indicated by the following abbreviations in the text. DCC: N,N'-dicyclohexylcarbodiimide HONB: N-hydroxy-5-norbornene-2,3-dicarboximide ONB: HONB ester Z: benzyloxycarbonyl BOC: t-butoxycarbonyl But: t-butyl DMF: dimethylformamide Bzl: Benzyl HOBT: N-Hydroxybenztriazole MeOH: Methyl alcohol AcOEt: Ethyl acetate TEA: Triethylamine THF: Tetrahydrofuran When amino acids or their residues are expressed in the above abbreviations herein, unless otherwise specified, L - means D- body, (D)- or D
- Specify. In the above general formulas () and (), R ₁
Examples of straight-chain or branched alkyl having 1 to 6 carbon atoms represented by include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-bepyl, isopentyl sec -amyl, tert-amyl, n-hexane, isohexane, 3-methylpentane, neohexane, 2,3-dimethylbutane and the like. Examples of side chains of D-α-amino acids represented by R ₂ and R′ ₂ include D-Len, D-Ala, D
-methionine, D-serine, D-threonine, D
-phenylalanine, D-α-amino-butyric acid, D
-valine, D-norvaline, D-norleucine,
Examples include side chains such as D-isoleucine. Examples of the alkyl having 1 or 2 carbon atoms represented by R ₃ include methyl and ethyl. The linear or branched alkyl having 1 to 8 carbon atoms represented by R ₄ is, for example, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl,
n-pentyl, isopentyl, sec-amyl,
tert-amyl, n-hexyl, isohexyl, 3
-methylpentyl, neohexyl, 2,3-dimethylbutyl, n-heptyl, iso-heptyl,
tert-heptyl, n-octyl, isooctyl,
Examples include tert-octyl. The linear or branched saturated aliphatic acyl having 1 to 8 carbon atoms represented by R ₄ is, for example, formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, valeryl, isovaleryl, hexanoyl, isohexanoyl. , heptanoyl, octanoyl, etc. Among them, those having 3 to 6 carbon atoms are particularly preferred. Examples of the halogen which may be substituted with the above groups include chlorine fluorine. Also,
Examples of C1-C4 alkoxy which may be substituted with the above groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-
Examples include butoxy, isobutoxy, tert-butoxy and the like. Among them, methoxy, ethoxy, n-
Propoxy is particularly preferred. In the above general formula, examples of the protecting group represented by Y ₁ include benzyloxycarbonyl, t-butoxycarbonyl, t-amyloxycarbonyl, isoniboryloxycarbonyl, adamantyloxycarbonyl, chloro- or nitro-substituted benzyloxy Examples include carbonyl, o-phenylthio, diphenylphosphinothioyl, and the like. Examples of the protecting group represented by Y ₂ include benzyl and t-butyl. Regarding the protecting groups for R′ ₂ and R′ ₄ , the above Y ₂
Examples of the protecting group for protecting the amino group of the basic group include those having the same meaning as Y ₁ above. Examples of the protecting group represented by Y ₃ include those having the same meaning as Y ₁ . In order to produce a peptide derivative having the chemical structure of the tetrapeptide derivative () of the present invention, partial amino acids that may constitute the polypeptide of the object () or the peptide and a compound that may constitute the remainder thereof are synthesized by peptide synthesis means. This is done by condensation. The peptide synthesis means may follow any known method, for example M. Bodansky and
Written by MAOndetti, Peptide Synthesis, Intef science, NeW
York, 1966; FMFinn and K. Hofmann, The Proteins, Volume 2, H.
Edited by Nenrath, RL Hill, Academic Press Inc.
NeW York, 1976; methods described in "Peptide Synthesis" by Nobuo Izumiya et al., Maruzen Co., Ltd., 1975, such as azide method, chloride method, acid anhydride method, mixed acid anhydride method, DCC method, active ester method. , a method using Woodward's reagent K, a carbodiimidazole method, a redox method, a DCC/HONB method, and the like. In some cases, the NCA method (N-carboxyanhydride; a method using an intramolecular cyclic carbonyl compound corresponding to an amino acid without using a protecting group) may be applied. Before carrying out the main condensation reaction, carboxyl groups that do not participate in the reaction of the raw materials are removed by means known per se.
The amino group may be protected, or the carboxyl group or amino group involved in the reaction may be activated. Examples of the protecting group for the raw material include the above-mentioned protecting groups. The carboxyl group of the raw material is, for example, a metal salt (e.g., sodium, potassium salt, etc.), a t-alkylamine salt (e.g., triethylamine, N-methylmorpholine, etc.), or an ester (e.g., methyl, ethyl, benzyl, p-nitrobenzyl). , t-butyl, t-amyl, etc.). As a protecting group for the amino group of the raw material,
For example, benzyloxycarbonyl group, t-butoxycarbonyl group, isobornyloxycarbonyl group, etc. are used as protecting groups for the imino group of histidine. Kishi etc. are mentioned. Examples of protective groups for the hydroxyl group of tyrosine include ethers such as benzyl and t-butyl; examples of protective groups for the guanidino group of arginine include nitro group, tosyl group, carbobenzoxy, isobonyloxycarbonyl, and adamantyloxycarbonyl. etc. are exemplified. Examples of raw materials with activated carboxyl groups include corresponding acid anhydrides, azides,
Active esters [alcohols (e.g., bentachlorophenol, 2,4,5-trichlorophenol,
2,4-dinitrophenol, dianomethyl alcohol, p-nitrophenol, N-hydroxy-5-norbornene-2,3-dicarboximide, N-hydroxysuccinimide, N-hydroxyphthalimide, N-hydroxybenztriazole) and esters]. As raw material with activated amino groups,
Examples include the corresponding phosphoamides. Assuming that the raw materials are A and B, examples of combinations of the above-mentioned states of the carboxyl and amino groups of A and B are shown in the table below.

【table】

[Table] This reaction can be carried out in the presence of a solvent. The solvent can be appropriately selected from those known to be usable in peptide condensation reactions. Examples include anhydrous or hydrous dimethylformamide, dimethyl sulfoxide, pyridine, chloroform, dioxane, dichloromethane, tetrahydrofuran, or a suitable mixture thereof. The reaction temperature is appropriately selected from the range known to be usable for peptide bond forming reactions,
Usually, the temperature is appropriately selected from the range of about -20°C to about 30°C. The precursor (protected peptide) of the compound of the present invention can also be easily produced by solid phase synthesis. The thus obtained protected compound represented by formula () is subjected to a protecting group elimination reaction. The reaction varies depending on the type of protecting group used, but
In any case, it is industrially advantageous to remove all protecting groups in one step without affecting peptide bonds. Therefore, this point should be taken into consideration beforehand when employing a protecting group. Typical examples of combinations of protecting group types and elimination conditions are shown in Table 1 below.

[Table] The desorption conditions shown in Table 1 are, for example,
Catalytic reduction using palladium black, palladium carbon, platinum, etc. as a catalyst, acid decomposition using trifluoroacetic acid, dilute hydrochloric acid, methanesulfonic acid, etc. In addition, reduction using sodium in liquid ammonia, trifluoromethanesulfonic acid, etc. Acid decomposition using a glacial acetic acid solution of hydrobromic acid, hydrogen fluoride, etc. can also be mentioned. These reactions are generally carried out at an appropriate temperature of -20°C to 40°C, but in acid decomposition it is effective to add a cation scavenger such as anisole, phenol, or thioanisole. Peptide derivatives produced in this way ()
After the reaction is completed, the peptide is collected by peptide separation means such as extraction, distribution, reprecipitation, recrystallization, column chromatography, etc. Peptide () can be obtained by forming a salt with organic acid, inorganic acid, etc. by conventional methods, but it is generally obtained as a salt with acetate, citrate, tartrate, etc.
Hydrochloride, sulfate, etc. are preferred. Examples of pharmacological experiments using the compound () of the present invention are listed below. Hot plate method: Ta:CF ₁ mice, 4 weeks old and weighing 18 to 22 g, were used. When animals are placed on a copper plate maintained at 55 ± 0.5°C, they respond by licking their hind paw soles and attempting to escape by jumping up due to the thermal stimulation. Mice that showed such a reaction within 20 seconds were selected and experiments were conducted with 10 mice per group. The sample was injected intravenously or subcutaneously, and 5, 10, 20, 30, 45, and 60 minutes later, the reaction time was measured in the same manner as above, and the non-treated group (control group)
compared with the reaction time of In order to avoid irreversible damage to the sole of the foot, the reaction time measurement was limited to 60 seconds. The analgesic effect (prolongation rate %) was calculated using the following formula, and a significant difference test (t-test) was performed to determine the effect. Analgesic effect (%) = reaction time after administration (seconds) -
Reaction time before administration (seconds) / 60 - reaction time before administration (seconds)
×100 The compound () of the present application was tested in the above test.
An analgesic effect with a prolongation rate of 40% or more with a peak duration of 5 to 20 minutes was observed at a dose of 200 μg or less, and in the case of particularly effective compounds, a clear analgesic effect was observed even at a minute dose of 20 μg or less. As mentioned above, the compound () of the present invention shows a clear analgesic effect at a dose in the range of about 1 to 10 mg/Kg by intravenous administration by the hot plate method using mice, and has a more pronounced analgesic effect than β-endorphin. It is useful as an analgesic because it shows excellent effects. Therefore, the compound () of the present invention and its pharmacologically acceptable acid addition salts are suitable for use in mammals, e.g.
mice, rats, rabbits, dogs, monkeys, humans, etc.)
It can be used as an analgesic for terminal cancer patients and other severe pain. The toxicity of the compound () of the present invention and its acid addition salts is extremely low, and there have been no cases of death even at doses of 200 mg/Kg, which are far higher than the medicinally effective dose. The compounds of the invention may be administered as free forms or as acid addition salts thereof. In the case of the free form of the compound (), the dosage is generally an appropriate amount in the range of 0.1 mg to 50 mg per kg of body weight. The dosage of the acid addition salt of the compound () of the present invention is generally determined as the amount of the free form of the compound () per 1 kg of body weight.
The appropriate amount ranges from 0.1mg to 50mg. The compound is primarily administered parenterally (eg, intravenous or subcutaneous injection, rectal administration), but may also be administered orally in some cases. In addition, continuous injection during surgery,
Intravenous fluids are particularly useful. Examples of dosage forms include injections, suppositories, and powders, but infusion preparations are also useful. Since this compound is stable as a substance, it can be stored as a solution in physiological saline, but it can also be prepared into a lyophilized ampoule by adding mannitol and sorbitol and dissolved at the time of use. For intravenous or subcutaneous injection, for example, in physiological saline at a concentration of 12.5 to 25 mg/ml.
It is preferable to use a solution dissolved so that The present invention will be explained in more detail with reference to Examples below. Cephadex used to purify the final product
LH-20 is a product of Pharmacia (Sweden). The purity of the produced compound was examined by thin layer chromatography using Merck Kieselgel 60F-254.
The developing solvent is shown below. Rf ¹ : Chloroform-methanol-acetic acid (9:
1:0.5) Rf ² : Acetate ethyl ester-pyridine-acetic acid-
Water (60:20:6:10) Example 1H-Tyr-(D)-Leu-Gly-Phe-NH
Production of -NH-(CH ₂ ) ₂ -OH () Production of Z-(D)-Leu-Gly-OH Dissolve 3.3 g of glycine in 20 ml of water, add 3.4 g of sodium biphosphate, and mix with Z-( D) Leu
−ONB17.0g (40mmol) in DMF solution (100
ml). Stir overnight at room temperature, then add DMF
was distilled off, and 200 ml of AcOEt and 50 ml of N-hydrochloric acid were added to the residue.
Add ml. The AcOEt layer is washed with water and then dried with anhydrous sodium sulfate. AcOEt is distilled off and the residue is crystallized with petroleum benzine. AcOEt - recrystallized with petroleum benzine; 9.8g
(75%). Melting point 110-111℃, [α] ²⁷ _D +24.3゜(C=
1.0, MeOH), Rf ¹ =0.42. Elemental analysis As C ₁₆ H ₂₂ O ₅ N ₂ Calculated value: C59.61, H: 6.88, N8.69 Actual value: C59.59, H6.85, N8.74 () Z−Tyr−(D)−Leu -Production of Gly-OH Z-(D)-Leu-Gly-OH7.0g (22 mmol)
Dissolve in 100ml of MeOH and perform catalytic reduction using palladium black as a catalyst. Filter off the catalyst and MeOH
is distilled off, the residue is suspended in a mixed solvent of THF-water (100ml-10ml), cooled, and 3ml of TEA is added dropwise to dissolve. Add 10.5 g of Z-Tyr-ONB to this and stir overnight at room temperature. Distill THF to residue
Add 200 ml of AcOEt and 25 ml of N-hydrochloric acid, wash the AcOEt layer with water, and dry with anhydrous sodium sulfate. AcOEt
is distilled off and the residue is solidified with ether.
Recrystallize from AcOEt; 8.2 g (78%). Melting point 179
−181°C, [α] ²⁷ _D +45.2° (C=0.5, MeOH), Rf ¹
=0.38 Elemental analysis As C ₂₅ H ₃₁ O ₇ N ₃ Calculated value: C61.68, H6.51, N8.64 Actual value: C61.84, H6.44, N8.66 () Z−Phe−NH−NH −CH ₂ −CH ₂ −OH
Production of Z-Phe-ONB 6.9 g (15 mmoles) and 2-hydrazinoethanol 1.14 g are dissolved in 50 ml of dioxane and stirred at room temperature for 3 hours. Dioxane is distilled off and the precipitated crystals are washed with ether and collected by filtration. Recrystallize from ethanol; 4.3 g (81%).
Melting point 149-150℃, [α] ²⁰ _D -8.8゜(C=0.5,
MeOH), Rf ¹ = 0.57. Elemental analysis As C ₁₉ H ₂₃ O ₄ N ₃ Calculated values: C63.85, H6.48, N11.76 Actual values: C63.93, H6.48, N11.61 () Z−Tyr−(D)−Leu −Gly−Phe−NH
Production of -NH-CH ₂ -CH ₂ -OH Z-Phe-NH-NH-CH ₂ -CH ₂ -OH570mg
(1.6 mmoles) is dissolved in 50 ml of MeOH, and catalytic reduction is performed using palladium black as a catalyst. The catalyst was filtered off, MeOH was distilled off, and the residue was dissolved in 5 ml of THF.
On the other hand Z-Tyr-(D)-Leu-Gly-OH727mg
Dissolve (1.5 mmoles) in 20 ml of THF, cool to 0°C, add 295 mg of HONB and 340 mg of DCC, and stir at 0°C for 3 hours. Insoluble materials are removed by filtration, and the filtrate is combined with the previously prepared THF solution of the amine component, and stirred overnight at room temperature. Distill the THF and remove the residue
Extract with 100ml of AcOEt and wash with water. AcOEt is distilled off and the precipitated gel-like substance is collected by filtration. Reprecipitate with ethanol-ether; 850 mg (83%). melting point
151°C, [α] ²⁰ _D +14.8° (C = 0.5, MeOH), Rf ¹ =
0.28. Elemental analysis As C ₃₆ H ₄₆ O ₈ N ₆ Calculated values: C62.59, H6.71, N12.17 Actual values: C61.96, H6.99, N11.81 () H-Tyr-(D)-Leu −Gly−Phe−NH
Production of -NH-CH ₂ -CH ₂ -OH Z-Tyr-(D)-Leu-Gly-Phe-NH-NH
500 mg of -CH ₂ -CH ₂ -OH is dissolved in 50 ml of MeOH and catalytic reduction is performed using palladium black as a catalyst. The catalyst was filtered off, MeOH was distilled off, and the residue was diluted with 50% N-acetic acid.
ml and filter out insoluble matter. The powder obtained by freeze-drying the filtrate was dissolved in a small amount of N-acetic acid water and applied to a Sephadex LH-20 column (2.5 x 120 cm).
Elute with aqueous N-acetic acid. Collect the elution fractions from 307 ml to 340 ml and freeze-dry; 310 mg (62%),
[α] ²⁰ _D +16.7° (C = 0.35, MeOH), Rf ² = 0.41, Amino acid analysis value (acid hydrolyzate): Gly1.05(1), Leu1.00
(1), Tyr0.71(1), Phe0.92(1), average recovery rate 75%. Example 2H-Tyr-(D)-Leu-Gly-Phe-NH
-Manufacture of _NH2 () Z-Tyr-(D)-Leu-Gly-Phe-
Production of OMe Z-Tyr-(D)-Leu-Gly-OH1.45g
Dissolve 590 mg of HONB in 50 ml of THF, cool to 0°C, add 680 mg of DCC, and stir for 4 hours. The precipitated urea body is filtered off and H-Phe-OMe hydrochloride is added to the filtrate.
Add 750 mg and 0.5 ml of TEA and stir overnight at room temperature.
THF was distilled off and the residue was extracted with 100ml of AcOEt.
Wash with 5% deionized water and dry with anhydrous sodium sulfate. AcOEt is distilled off and hardened with petroleum benzine.
AcOEt - reprecipitated with ether; 1.73 g (89%). Melting point 100-101℃, [α] ²² _D -2.6゜ (C=0.5, DMF),
^Rf1 =0.68. Elemental analysis As C ₃₅ H ₄₂ O ₈ N ₄ Calculated values: C65.00, H6.55, N8.66 Actual values: C64.97, H6.95, N8.62 () Z−Tyr−(D)−Leu −Gly−Phe−NH
-Production of _NH2 Z-Tyr-(D)-Leu-Gly-Phe-OMe517mg
Dissolve in 5 ml of DMF, add 0.1 ml of NH ₂ -NH ₂ .H ₂ O, and let stand at room temperature for 3 days. DMF is distilled off, water is added to the residue, and filtered. Reprecipitate with MeOH-ether; 410 mg (80%). Melting point 176−
177°C (decomposition), [α] ²² _D −22.7° (C=0.4, DMF),
^Rf1 =0.20. Elemental analysis C ₃₄ H ₄₂ O ₇ N ₆・1/2H ₂ O Calculated value: C62.27; H6.45, N12.82 Actual value: C62.03, H6.57, N12.64 () H-Tyr -(D)-Leu-Gly-Phe-NH
-Production of _NH2 Z-Tyr-(D)-Leu-Gly-Phe-NH-
Dissolve 350 mg of NH ₂ in 50 ml of MeOH, add 0.1 ml of acetic acid, and perform catalytic reduction using palladium black as a catalyst. The catalyst was filtered off, the filtrate was concentrated, and the residue was dissolved in a small amount of aqueous N-acetic acid, applied to a Sephadex LH-20 column (2.5 x 120 cm), and eluted with aqueous N-acetic acid.
Collect the elution fractions from 320 ml to 340 ml and freeze-dry; 210 mg (60%), [α] ²² _D + 22.6° (C - 0.38
，
MeOH), ^Rf2 = 0.34. Amino acid analysis value (acid hydrolyzate): Gly1.00(1), Leu1.00(1), Tyr0.90(1),
Phe1.02(1) (average recovery rate 70%). Example 3 H-Tyr-(D)-Ala-Gly-Phe-
Production of NH- _NH2 () Production of Z-Tyr-(D)-Ala-Gly-OH Z-(D)-Ala-Gly-OBu ^t 8.2g was added to MeOH200
ml and perform catalytic reduction using palladium black as a catalyst. The catalyst was filtered off, MeOH was distilled off, and the residue was dissolved in 70 ml of trifluoroacetic acid and left at room temperature for 40 minutes. Trifluoroacetic acid was distilled off and the residue was filtered with ether. Dissolve this powder in 50ml of water and add 4.7g of bicarbonate of soda. 13.3 g (28 mmol) of Z-Tyr-ONB was added to this solution.
Add 100 ml of THF solution and stir overnight at room temperature. Add N-hydrochloric acid to the reaction solution to neutralize it, and add 150ml of AcOEt.
Extract with Wash with water and dry with anhydrous sodium sulfate. AcOEt is distilled off and filtered with ether. Recrystallize from AcOEt; 8.5 g (69%). Melting point 184-185℃, [α] ²² _D +25.6゜(C=0.5, MeOH),
^Rf1 =0.16. Elemental analysis As C ₂₂ H ₂₅ O ₇ N ₃ Calculated values: C59.58, H5.68, N9.48 Actual values: C59.29, H5.81, N9.32 () Production of Z-Phe-NHNH-BOC Z-Phe-ONB18.4g and _NH2 -NH-BOC5.3
Dissolve g in 100ml of THF and stir overnight at room temperature.
THF was distilled off and the residue was extracted with 150ml of AcOEt.
Wash with citric acid water and 5% dehydrated water. Anhydrous Na ₂ SO ₄
and evaporate AcOEt. Filter it with petroleum benzene. AcOEt - recrystallized with petroleum benzine; 13.2 g (80%). Melting point 107-109℃, [α]
_19D −23.4° (C=0.5, MeOH), Rf ¹ =0.71. Elemental analysis As C ₂₂ H ₂₇ O ₅ N ₃ Calculated values: C63.90, H6.58, N10.16 Actual values: C64.18, H6.61, N9.88 () Z−Tyr−(D)−Ala −Gly−Phe−NH
-Production of NH-BOC 3.6 g of Z-Phe-NH-NH-BOC was added to 100 MeOH
ml, add 0.5 ml of acetic acid, and perform catalytic reduction using palladium black as a catalyst. Filter off the catalyst and make the filtrate
Add 10 ml of DMF and evaporate MeOH. On the other hand Z-
Tyr-(D)-Ala-Gly-OH3.5g and HONB1.56
Dissolve g in THF50ml and cool to 0℃ to DCC1.80.
Add g and stir at 0℃ for 6 hours. Insoluble materials are filtered off, and the filtrate is combined with the amine component and stirred overnight at room temperature. Distill the THF and remove the residue
Extract with 150ml of AcOEt and wash with 5% deuterated water and citric acid water. Dry with anhydrous sodium sulfate,
AcOEt is distilled off and hardened with petroleum benzene. AcOEt - recrystallized with petroleum benzine; 3.7g
(67%). Melting point 143-146℃ (decomposition), [α] ²² _D −25.4
° (C=0.5, DMF), Rf ¹ =0.38. Elemental analysis As C ₃₆ H ₄₄ O ₉ N ₆ Calculated value: C61:35, H6.29, N11.93 Actual value: C61.05, H6.42, N11.58 () Z−Tyr−(D)−Ala −Gly−Phe−NH
-Production of _NH2 Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
- Dissolve 3.5 g of BOC in 30 ml of trifluoroacetic acid and leave at room temperature for 20 minutes. Trifluoroacetic acid was distilled off and the residue was filtered with ether. Dissolve this powder in 10ml of DMF, add 1.4ml of TEA and stir. DMF is distilled off and the residue is collected by filtration with water. Reprecipitate with methanol to ether; 2.8
g (96%). Melting point 182-184℃ (decomposition), [α] ²² _D −
23.0 (C=0.5, DMF), Rf ¹ = 0.17 Elemental analysis As C ₃₁ H ₃₆ O ₇ N ₆ Calculated value: C61.58, H6.00, N13.90 Actual value: C61.32, H5.91, N13 .72 () H-Tyr-(D)-Ala-Gly-Phe-NH
-Production of _NH2 Z-Tyr-(D)-Ala-Gly-Phe-NH-
Dissolve 250 mg of NH ₂ in 50 ml of MeOH, add 0.1 ml of acetic acid, and perform catalytic reduction in the presence of palladium black. The catalyst was filtered off, and the residue was dissolved in a small amount of N-acetic acid water, applied to a Sephadex LH-20 column (2.5 x 120 cm), and eluted with N-acetic acid water. Collect the elution fractions from 310 ml to 335 ml and freeze-dry; 95 mg (38
%), [α] ²² _D +20.5° (C=0.2, MeOH), Rf ² =
0.22, Amino acid analysis value (hydrochloric acid hydrolyzate): Gly1.00
(1), Ala1.05(1), Tyr0.89(1), Phe1.01(1) (average recovery rate 81%). Example 4H-Tyr-(D)-Leu-Gly-Phe-NH
-NH-CO-CH ₃ production () Z-Phe-NH-NH-CO-CH ₃ production Z-Phe-NH-NH ₂ 2.03g (6.5 mmol)
Dissolve in 50ml of THF, cool and add 1.4ml of acetic anhydride.
Drop 0.9ml of TEA. Stir at room temperature for 2 hours. The precipitated crystals are collected by filtration and washed with ether.
Recrystallize from AcOEt; 1.9 g (83%). melting point 205
−206°C, [α] ²³ _D −16.4° (C=0.5, DMF), Rf ¹ =
0.60 Elemental analysis As C ₂₀ H ₂₃ O ₅ N ₃ Calculated value: C62.32, H6.02, N10.90 Actual value: C62.38, H6.09, N10.49 () Z−Tyr−(D)− Leu−Gly−Phe−NH
-NH-CO- _CH3 production Z-Phe-NH-NH-CO- _CH3 700mg
Dissolve in 50ml of MeOH, add 0.2ml of acetic acid, and perform catalytic reduction using palladium black as a catalyst. filter off the catalyst
Distill MeOH and dissolve the residue in 20 ml of THF. On the other hand, 680 mg (1.4 mmol) of Z-Tyr-(D)-Leu-Gly-OH and 300 mg of HONB were dissolved in 20 ml of THF, cooled, and 330 mg of DCC was added, followed by stirring at 0°C for 5 hours. Insoluble materials are removed by filtration, and the filtrate is combined with the amine component prepared earlier and stirred overnight at room temperature.
THF is distilled off, AcOEt is added to the residue, and the mixture is filtered. Reprecipitate with methanol to AcOEt; 0.71
g (74%). Melting point 184-185℃, [α] ^23D _-13.4゜(C
=
Dissolve to 0.50 ml, add 0.1 ml of acetic acid, and perform catalytic reduction in the presence of palladium black. Filter off the catalyst, dissolve the residue in a small amount of N-acetic acid water, and add Cephadex LH.
-20 column (2.5 x 120 cm) and elute with N-acetic acid water. Collect the elution fractions from 310 ml to 335 ml and freeze-dry; 95 mg (38%), [α] ²² _D + 20.5° (C = 0.2, MeOH), Rf ² = 0.22, amino acid analysis value (hydrochloric acid hydrolyzate) ): Gly1.00(1), Ala1.05(1), Tyr0.89
(1), Phe1.01(1) (average recovery rate 81%). Example 4H-Tyr-(D)-Leu-Gly-Phe-NH
-NH-CO-CH ₃ production () Z-Phe-NH-NH-CO-CH ₃ production Z-Phe-NH-NH ₂ 2.03g (6.5 mmol)
Dissolve in 50ml of THF, cool and add 1.4ml of acetic anhydride.
Drop 0.9ml of TEA. Stir at room temperature for 2 hours. The precipitated crystals are collected by filtration and washed with ether.
Recrystallize from AcOEt; 1.9 g (83%). melting point 205
−206°C, [α] ²³ _D −16.4° (C=0.5, DMF), Rf ¹ =
0.60 Elemental analysis As C ₂₀ H ₂₃ O ₅ N ₃ Calculated value: C62.32, H6.02, N10.90 Actual value: C62.38, H6.09, N10.49 () Z−Tyr−(D)− Leu−Gly−Phe−NH
-NH-CO- _CH3 production Z-Phe-NH-NH-CO- _CH3 700mg
Dissolve in 50ml of MeOH, add 0.2ml of acetic acid, and perform catalytic reduction using palladium black as a catalyst. filter off the catalyst
Distill MeOH and dissolve the residue in 20 ml of THF. On the other hand, 680 mg (1.4 mmol) of Z-Tyr-(D)-Leu-Gly-OH and 300 mg of HONB were dissolved in 20 ml of THF, cooled, and 330 mg of DCC was added, followed by stirring at 0°C for 5 hours. Insoluble materials are removed by filtration, and the filtrate is combined with the previously prepared amine component and stirred overnight at room temperature.
THF is distilled off, AcOEt is added to the residue, and the mixture is filtered. Reprecipitate with methanol to AcOEt; 0.71
g (74%). Melting point 184-185℃, [α] ^23D _-13.4゜(C
=
0.5, DMF), Rf ¹ = 0.28 Elemental analysis As C ₃₆ H ₄₄ O ₈ N ₆ Calculated value: C62.77, H6.44, N12.20 Actual value: C62.52, H6.81, N12, 04 () H-Tyr-(D)-Leu-Gly-Phe-NH
-NH-CO- _CH3 production Z-Tyr-(D)-Leu-Gly-Phe-NH-NH
-CO-CH ₃ 500mg is dissolved in MeOH50ml and acetic acid 1
ml and perform catalytic reduction using palladium black as a catalyst. The catalyst is removed by filtration, the filtrate is distilled off, water is added to the residue, and insoluble matter is removed by filtration. The powder obtained by freeze-drying the filtrate is dissolved in a small amount of N-acetic acid water and then
Apply to LH-20 column (2.5 x 120 cm) and elute with N-acetic acid water. Elution fractions from 320 ml to 345 ml are collected and lyophilized; 245 mg (50%). [α] ²³ _D
+26.7° (C=0.3, MeOH), Rf ² = 0.41, Amino acid analysis value (hydrochloric acid hydrolyzate): Gly1.00(1), Leu1.05(1),
Tyr0.93(1), Phe1.02(1) (average recovery rate 82%). Example 5 H-Tyr-(D)-Ala-Gly-Phe-
Production of NH-NH-CO-( _CH2 ) _{3 -CH3} () Z-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO-( _CH2 ) _{3 -CH3} Production Z-Tyr-(D)-Ala-Gly-Phe-NH-
NH ₂ 604 mg (1 mmol) and HOBT 297 mg, CH ₃
Dissolve 0.2 ml of (CH ₂ ) ₃ COOH in 5 ml of DMF, cool to 0°C, add 454 mg of DCC, and stir at 0°C for 4 hours at room temperature overnight. Insoluble matter was filtered off, DMF was distilled off, and the residue was extracted with 100 ml of AcOEt and washed with 5% dehydrated water. Dry over anhydrous sodium sulfate, remove AcOEt, and filter the precipitated gel with ether. Reprecipitate with methanol to AcOEt; 480mg
(70%). Melting point 178-179℃, [α] ²³ _D -22.4゜(C=
0.16, DMF), Rf ¹ = 0.46 Elemental analysis As C ₃₆ H ₄₄ O ₈ N ₆ Calculated value: C62.77, H6.44, N12.20 Actual value: C62.71, H6.77, N11.98 () H-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO-( _CH2 ) _{3- CH3} Production Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
Dissolve 430 mg of -CO-(CH ₂ ) ₃ -CH ₃ in 50 ml of MeOH, add 1 ml of acetic acid, use palladium black as a catalyst,
Perform catalytic reduction. The catalyst was filtered off, MeOH was distilled off, and the residue was dissolved in a small amount of N-acetic acid water and applied to a Sephadex LH-20 column (2.5 x 120 cm).
- Elute with aqueous acetic acid. Elution fractions from 325 ml to 340 ml are collected and lyophilized; 240 mg (56%).
[α] ²³ _D +24.5° (C = 0.2, MeOH), Rf ² = 0.47, Amino acid analysis value (hydrochloric acid hydrolyzate): Gly1.00(1),
Ala0.95(1), Tyr0.88(1), Phe1.01(1) (average recovery rate
82%) Example 6 H-Tyr-(D)-Ala-Gly-Phe-
Production of NH-NH-CO- _CH2 -CH-( _CH3 ) ₂ () Z-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO- _CH2 -CH-( _CH3 ) ₂ Production Z-Tyr-(D)-Ala-Gly-Phe-NH-
530 mg (0.88 mmol) of NH ₂ and (CH ₃ ) ₂ −CH−
Prepared as in Example 5 () using 0.15 ml of CH ₂ -COOH; 320 mg (53%). melting point
228−230°C, [α] ²³ _D −23.9° (C=0.28, DMF),
Rf ¹ = 0.46 Elemental analysis As C ₃₆ H ₄₄ O ₈ N ₆ Calculated values: C62.77, H6.44, N12.20 Actual values: C62.58, H6.35, N11.95 () H−Tyr−( D) -Ala-Gly-Phe-NH
-NH-CO- _CH2 -CH-( _CH3 ) ₂ Production Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
Example 5 using 170 mg of -CO-CH ₂ -CH-(CH ₃ ) ₂
Prepare in the same manner as for (); 105mg
(62%). [α] ²³ _D +21.1° (C=0.18, MeOH), Rf ²
=0.49, Amino acid analysis value (hydrochloric acid hydrolyzate): Gly1.00
(1), Ala1.02(1), Tyr0.96(1), Phe1.02(1) (average recovery rate 80%). Example 7 H-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO-( _CH2 ) ₂ -Cl production () Z-Tyr-(D)-Ala-Gly-Phe-NH
Production of -NH-CO-( _CH2 ) ₂ -Cl Z-Tyr-(D)-Ala-Gly-Phe-NH-
513 mg (0.85 mmol) of _NH2 and Cl− _CH2 − _CH2−
Prepared as in Example 5 () using 120 mg of COOH; 360 mg (59%). Melting point 169-170℃,
[α] ²³ _D −21.1° (C=0.37, DMF) Elemental analysis C ₃₄ H ₃₉ O ₈ N ₆・Cl Calculated value: C58.74, H5.65, N12.09, Cl5.10 Actual value: C58. 61, H5.59, N11.81, Cl4.92 () H-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO-( _CH2 ) ₂ -Cl production Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
250 mg of -CO-( _CH2 ) ₂ -Cl are prepared as in Example 5 (); 150 mg (60%). [α]
²³ _D +20.5° (C=0.2, MeOH), Rf ² =0.42. Amino acid analysis value (acid hydrolyzate): Gly1.01(1), Ala0.95(1),
Tyr0.88(1), Phe0.98(1) (average recovery rate 79%). Example 8H-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO- _CH2 - _CH2 -OH production () Z-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO- _CH2 - _CH2 -OH Production Z-Tyr-(D)-Ala-Gly-PheNH-- _NH2
Using 540 mg and 0.12 ml of HO−CH ₂ −CH ₂ −COOH,
Prepared in the same manner as in Example 5 (); 320 mg
(53%). Melting point 178-179℃, [α] ²³ _D -22.3゜(C=
0.3, DMF), Rf ¹ = 0.10 Elemental analysis As C ₃₄ H ₄₀ O ₉ N ₆ Calculated values: C60.34, H5.96, N12.42 Actual values: C59.98, H5.85, N12.18 () H-Tyr-(D)-Ala-Gly-Phe-NH
Production of -NH-CO-CH ₂ -CH ₂ -OH Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
The desired product was obtained from 250 mg of -CO-CH ₂ -CH ₂ -OH in the same manner as in () of Example 5; 100 mg (40%),
[α] ²³ _D +19.5° (C = 0.21, MeOH), Rf ² = 0.32 Amino acid analysis value (acid hydrolyzate): Gly1.00(1), Ala1.05(1),
Tyr0.92(1), Phe1.00(1) (average recovery rate 79%). Example 9H-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO- _CH2 - _CH3 production () Z-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO- _CH2 - _CH3 Production Z-Tyr-(D)-Ala-Gly-Phe-NH- _NH2
495 mg (0.82 mmol) and _CH3 − _CH2 −COOH0.1
350 mg (65%) melting point 190-191°C, [α] ²³ _D −
20.5° (C=0.3, DMF), Rf ¹ =0.26. Elemental analysis C ₃₄ H ₄₀ O ₈ N ₆ Calculated values: C61.80, H6.10, N12.72 Actual values: C61.53, H6.02, N12.65 () H-Tyr-(D)-Ala −Gly−Phe−NH
-NH-CO-CH2- _H3 Production Z-Tyr-( _D )-Ala-Gly-Phe-NH-NH
The desired product was obtained using 250 mg of -CO-CH ₂ -CH ₃ in the same manner as in Example 5 (). ;130mg (52%), [α] ²³ _D
+25.5° (C=0.23, MeOH), Rf ² =0.42. Amino acid analysis value (acid hydrolyzate): Gly1.00(1), Ala0.95(1),
Phe1.01(1), Tyr0.95(1) (average recovery rate 81%). Example 10H-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO- _CH2 - _CH2 - _NH2 production () Z-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO- _CH2 - _CH2 -NH-Z Production Z-Tyr-(D)-Ala-Gly-Phe-NH- _NH2
495 mg (0.82 mmols) and Z-β-Ala-ONB 390 mg
Dissolve in 10ml of DMF, cool, add 0.14ml of TEA, and stir overnight at room temperature. DMF is distilled off and the residue is extracted with 100 ml of AcOEt, washed with 5% deuterated water and dried over anhydrous sodium sulfate. AcOEt is distilled off and the residue is filtered off with ether. Reprecipitate with methanol to AcOEt; 370 mg (56%). melting point
231−233°C, [α] ²³ _D −21.5° (C=0.27, DMF),
^Rf1 =0.26. Elemental analysis C ₄₂ H ₄₇ O ₁₀ N ₇ Calculated value: C62.28, H5.85, N12, 10 Actual value: C61, 99, H5.94, N11.95 () H-Tyr-(D)-Ala- Gly−Phe−NH
-NH-CO- _{CH2CH2 - NH2} Production Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
-CO-CH ₂ -CH ₂ -NH-Z 280 mg was treated in the same manner as in Example 5 () to obtain the target product; 160 mg (61
%). [α] ²³ _D +16.7° (C=0.27, MeOH), Rf ² =0.07,
Amino acid analysis value: Gly1.00(1), Ala0.95(1), β-
Ala0.99(1), Tyr0.89(1), Phe1.00(1) (average recovery rate
79%). Example 11H-Tyr-(D)-Ala-Gly-Phe-NH
Production of −NH−CO−(CH ₂ ) ₂ −CH ₃ () Production of Z-Phe-NH-NH ₂ 2.5g (8mmoles)
Dissolve in 30 ml of DMF and cool to CH ₃ CH ₂ CH ₂ COCl0.9
ml and TEA1.1ml and stir at room temperature for 5 hours.
Water is added to the reaction solution, and the precipitated crystals are collected by filtration and dried. AcOEt - recrystallized from petroleum benzine;
2.6g (85%). Melting point 150-156°C, [α] ²³ _D -27.2° (C = 0.5, MeOH), Rf ¹ = 0.71. Elemental analysis As C ₂₁ H ₂₅ O ₄ N ₃ Calculated values: C65.46, H6.48, N10.87 Actual values: C65.78, H6.57, N10.96 () Z−Tyr−(D)−Ala −Gly−Phe−NH
Production of -NH-CO-CH ₂ -CH ₂ -CH ₃ Z-Phe-NH-NH-CO-CH ₂ -CH ₂ -CH ₃
Dissolve 505 mg in 50 ml of MeOH and perform catalytic reduction using palladium black as a catalyst. Filter off the catalyst and MeOH
was distilled off and the residue was dissolved in 20 ml of THF. On the other hand Z-
Tyr-(D)-Ala-Gly-OH530mg and HONB237
Dissolve mg in THF50ml, cool to 0℃, and DCC271mg.
Add and stir at 0℃ for 4 hours. The precipitated urea body is removed by filtration, and the filtrate is combined with the previously prepared amine component and stirred overnight at room temperature. THF is distilled off and the residue is extracted with 100 ml of AcOEt, washed with 5% deuterated water and dried over anhydrous sodium sulfate.
AcOEt is distilled off and the residue is filtered off with ether. Reprecipitate from AcOEt; 420 mg (50%).
Melting point 203-206℃, [α] ²³ _D -0.4゜(C=0.5,
MeOH), Rf ¹ = 0.28. Elemental analysis As C ₃₅ H ₄₂ O ₈ N ₆ Calculated values: C62.30, H6.27, N12.46 Actual values: C62.60, H6.52, N12.18 () H-Tyr-(D)-Ala −Gly−Phe−NH
-NH-CO-CH ₂ CH ₂ CH ₃ production Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
Dissolve 250 mg of -CO-CH ₂ -CH ₂ -CH ₃ in 50 ml of MeOH, add 0.2 ml of acetic acid, and perform catalytic reduction using palladium black as a catalyst. The catalyst was filtered off, the filtrate was concentrated, and the residue was dissolved in a small amount of N-acetic acid water, applied to a Sephadex LH-20 column (2.5 x 125 cm), and eluted with N-acetic acid water. Collect the elution fractions from 320 ml to 340 ml and freeze-dry; 110 mg (44
%). [α] ²³ _D +23.2° (C=0.25, MeOH), Rf ² =
0.40, Amino acid analysis (hydrochloric acid hydrolyzate): Gly1.06(1),
Ala1.00(1), 0.93(1), Phe0.93(1) (average recovery rate 90
%). Example 12H-Tyr-(D)-Ala-Gly-Phe-NH
Production of −NH−CO−OCH ₂ −CH ₃ () Z−Phe−NH−NH−CO−OCH ₂ −
Production of CH ₃ Example 11 () using 1.8 g (5.7 mmol) of Z-Phe-NH-NH ₂ and 0.7 ml of ethyl chlorocarbonate.
Synthesized in the same manner as; 1.5g (69%). melting point 134
°C, [α] ²³ _D −20.6° (C = 0.5, DMF), Rf ¹ = 0.69. Elemental analysis As C ₂₀ H ₂₃ O ₅ N ₃ Calculated value: C62.32, H6.02, N10.90 Actual value: C62.38, H6.09, N10.49 () Z−Tyr−(D)−Ala −Gly−Phe−NH
-NH-CO- _OCH2 - _CH3 Production Z-Tyr-(D)-Ala-Gly-OH580mg (1.3 mmol) and Z-Phe-NH-NH-CO- _OCH2-
Synthesize in the same manner as in Example 11 () using 620 mg of CH ₃ ; 820 mg (93%), melting point 154-155°C, [α]
²³ _D −24.2° (C = 0.45, DMF), Rf ¹ = 0.40. Elemental analysis C ₃₄ H ₄₀ O ₉ N ₆ Calculated values: C60.34, H5.96, N12.42 Actual values: C60.09, H5.87, H12.21 () H-Tyr-(D)-Ala −Gly−Phe−NH
Production of -NH-CO-OCH ₂ -CH ₃ Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
-CO-OCH ₂ -CH ₃ 450 mg () of Example 11
Treat in the same manner as to obtain the desired product; 240mg
(53%), [α] ²³ _D +18.1° (C = 0.2, MeOH), Rf ²
=
0.49. Amino acid analysis value: Gly1.00(1), Ala0.98(1),
Tyr0.88(1), Phe1.02(1), (average recovery rate 78%). Example 13 H-Tyr-(D)-Ala-Gly-MePhe-NH-
Production of NH ₂ () Z−Tyr−(D)−Ala−Gly−MePhe−
Production of OCH ₃ Z-MePhe-OH1.8g (5.7 mmol)
Dissolve in 20ml of MeOH, 5N-hydrochloric acid/dioxane 2
ml and leave at room temperature overnight. The solvent was distilled off, and the residue was extracted with 100 ml of AcOEt, washed with water, and dried over anhydrous sodium sulfate. 1.5g after distilling off AcOEt
Remove the oily substance. 1.1 g of this was dissolved in 50 ml of MeOH and subjected to catalytic reduction using palladium black as a catalyst. Filter off the catalyst, distill off MeOH, and remove the residue.
Dissolve in 20ml of DMF. To this, Z-Tyr-(D)-Ala
-Gly-OH1.55g (3.5mmol) and HONB0.75
g, cooled to 0℃, added 0.80g of DCC, and cooled to 0℃.
Stir overnight at room temperature for 5 hours. The precipitated urea body is filtered off and DMF is distilled off. AcOEt100 residue
ml and wash with 5% dehydrated water. Dry over anhydrous sodium sulfate, distill off AcOEt, and solidify with ether. Reprecipitate with MeOH-ether; 1.2
g (56%). Melting point 103-104℃, [α] ^23D _-17.2゜(C
=
0.31, MeOH), Rf ¹ =0.68. Elemental analysis As C ₃₃ H ₃₈ O ₈ N ₄ Calculated values: C64.06, H6.19, N9.06 Actual values: C63.91, H6.05, N8.89 () Z−Tyr−(D)−Ala −Gly−MePhe−
Production of NH- _NH2 Z-Tyr-(D)-Ala-Gly-MePhe-
OCH ₃ 1.0 g (1.6 mmol) of Example 2 ()
Treat with 0.5 ml of NH ₂ -NH ₂ -H ₂ O in the same manner as above to obtain the desired product; 820 mg (82%). Melting point 95-96℃ (decomposition), [α] ²³ _D −26.8゜ (C = 0.5, DMF), Rf ¹ = 0.29
. Elemental analysis As C ₃₂ H ₃₈ O ₇ N ₆ Calculated values: C62.12, H6.19, N13.59 Actual values: C61.85, H6.28, N13.09 () H-Tyr-(D)-Ala −Gly−MePhe−
Production of NH- _NH2 Z-Tyr-(D)-Ala-Gly-MePhe-NH-
300 mg of NH ₂ was treated in the same manner as in Example 5 () to obtain the target product; 155 mg (52%), [α] ²³ _D +12.0° (C = 0.2, MeOH), Rf ² = 0.46, amino acid. Analysis value (acid hydrolyzate): Gly1.00(1), Ala0.98(1), Tyr0.91(1)
(Average response rate 79%). Example 14 H-Tyr-(D)-Ala-Gly-Phe-NHNH-
Production of COCH ₂ −CH ₂ −CH ₂ −Cl () Z−Phe−NHNH−COCH ₂ −CH ₂ −
Production of CH ₂ -Cl Z-Phe-NHNH ₂ 3.13g (10mmoles) and γ
- Dissolve 1.27 g (10 mmoles) of chlorobutyric acid in 40 ml of DMF, cool to 0°C, add 2.2 g of DCC, and stir at room temperature for 20 hours. After filtering the reaction solution, it was dried under reduced pressure, and the residue was dissolved in 150 ml of ethyl acetate.
Wash with N-hydrochloric acid solution and 4% sodium bicarbonate solution, dry over anhydrous sodium sulfate, and dry under reduced pressure. Crystallization from ethyl acetate-petroleum ether gives 3.6 g (85.7%) of needles. Melting point 185−186
℃. Rf ¹ = 0.32 Elemental analysis C ₂₁ H ₂₄ O ₄ N ₂ Cl Calculated value: C60.35, H5.79, N10.06, Cl 8.49 Actual value: C60.51, H5.60, N10.15, Cl 8.27 () H-Tyr-(D)-Ala-Gly-Phe-
Production of NHNH−COCH ₂ CH ₂・Cl Z−Phe−NHNHCOCH ₂ −CH ₂ −CH ₂ −Cl
420 mg was catalytically reduced in 50 ml of methanol using palladium black as a catalyst. The catalyst was filtered off, the filtrate was dried under reduced pressure, and the residue and Z-Tyr-(D)-Ala-Gly
−Dissolve 445mg of OH in 10ml of DMF and add
Add 180mg of HONB and cool to O℃.
Add 250mg of DCC. After stirring at 0°C for 3 hours and at room temperature for 12 hours, insoluble materials were filtered off and DMF was added under reduced pressure.
to remove. This was dissolved in 100 ml of a 1:2 mixture of n-butanol and ethyl acetate, and mixed with N-hydrochloric acid and 4
Wash with % sodium bicarbonate water. Wash with water, remove the solvent under reduced pressure, dissolve in 50 ml of methanol,
ml of glacial acetic acid is added for catalytic reduction using palladium black as a catalyst. The catalyst was filtered off and dried under reduced pressure, and the residue was dissolved in 10 ml of N-acetic acid water. Filter off the insoluble matter and pour it into a Cephadex LH-20 column (3 x 45 cm), and flush with N-acetic acid water. The target substance flows out in the 215 ml to 236 ml section, which is collected and freeze-dried. You get 360mg of the desired product. ^Rf2 =0.485,
[α] ²³ _D +18.2° (C=0.28, MeOH), Amino acid analysis: Gly1.00(1), Ala1.02(1), Tyr0.89(1), Phe1.00
(1), average recovery rate 78%. Example 15 H-Tyr-(D)-Ala-Gly-Phe-
Production of NH-NH-CO-CH(OH) _-CH3 () Z-Tyr-(D)-Ala-Gly-Phe-NH
-NH-CO-CH(OH) _-CH3 Production Z-Tyr-(D)-Ala-Gly-Phe- _NHNH2
The desired product was obtained in the same manner as in Example 5 () using 510 mg and 0.1 ml of lactic acid; 420 mg (73%). melting point 134
−135℃, Rf ¹ =0.24, [α] ²⁴ _D −21.7゜(C=
0.7DMF). Elemental analysis C ₃₄ H ₄₀ O ₉ N ₆・1/2H ₂ O Calculated value: C=59.55, H6.02, N12.25 Actual value: C59.74, H6.29, N12, 47 () H-Tyr -(D)-Ala-Gly-Phe-NH
Production of -NH-COCH(OH) _-CH3 Z-Tyr-(D)-Ala-Gly-Phe-NH-NH
300 mg of -CO-CH(OH)-CH ₃ was deprotected and purified in the same manner as in Example 5 () to obtain the desired product (160 mg).
Get mg. Rf ² =0.34, [α] ²⁴ _D +16.7° (C=
0.45MeOH). Amino acid analysis: Gly0.91(1), Ala 1.00(1), Tyr0.97
(1), Phe1.03(1) (average recovery rate 81%) Example 16 H-Tyr-(D)-Thr-Gly-Phe-
Production of NH-NH-CO-( _CH2 ) ₃ - _CH3 (Thr=
Threonine) () Production of Z-(D)-Thr-Gly-OBu ^t Dissolve 5.06 g (20 mmol) of Z-(D)-Thr-OH and 3.60 g of H-Gly-OBu ^t in 50 ml of THF and heat at 0°C.
Cool to 0. Add 3.94g of HONB and 4.54g of DCC to 0.
Stir at room temperature for 3 hours at °C overnight. Insoluble materials are filtered off, THF is distilled off, and the residue is extracted with 100 ml of AcOEt, washed with 5% deuterated water, and dried over anhydrous sodium sulfate. AcOEt is distilled off and crystallized with petroleum benzene. AcOEt ~ Recrystallize with petroleum benzine; 5.0g
(69%). Melting point 56-57℃, [α] ²³ _D +13.8゜(C=
0.5MeOH), Rf ¹ = 0.69 Elemental analysis As C ₁₈ H ₂₆ O ₆ N ₂ Calculated values: C59.00, H7.15, N7.65 Actual values: C58.73, H7.11, N7.87 () Z Production of -Tyr-(D)-Thr-Gly-OH 4.6 g (12.5 mmol) of Z-(D)-Thr-Gly-OBu ^t is dissolved in 100 ml of MeOH and subjected to catalytic reduction using palladium black as a catalyst. Filter off the catalyst and MeOH
was distilled off and the residue was dissolved in 50 ml of THF. Z to this
Add 5.9 g of -Tyr-ONB and stir overnight at room temperature.
THF was distilled off and the residue was extracted with 100ml of AcOEt.
Wash with 5% deionized water and dry with anhydrous sodium sulfate. AcOEt is distilled off, and the precipitated gel-like substance is collected by filtration and dried (3.9 g). Dissolve 3.7 g of this in 40 ml of trifluoroacetic acid and leave at room temperature for 40 minutes. Trifluoroacetic acid was distilled off and the residue was filtered with ether. Recrystallize from AcOEt;
2.9g (87%). Melting point 143-144℃, [α] ²³ _D +18.2゜ (C = 0.5MeOH), Rf ² = 0.49 Elemental analysis C ₂₃ H ₂₇ O ₈ N ₃・1/2H ₂ O Calculated value C57.25, H5 .85, N8.87 Actual value C57.57, H5.92, N8.55 () Z−Tyr−(D)−Thr−Gly−Phe−
Production of OCH ₃ Dissolve 1.5 g (3.1 mmol) of Z-Tyr-(D)-Thr-Gly-OH and 0.63 g of HONB in 50 ml of THF.
Cool to 0°C, add 0.72g of DCC, and stir at 0°C for 3 hours. H-Phe-OCH ₃ hydrochloride in the reaction solution
Add 0.73g and 0.49ml of TEA and stir overnight at room temperature. Filter off the insoluble matter, distill off the THF, and remove the residue.
Extract with 100 ml of AcOEt, wash with 5% deuterated water and N-hydrochloric acid, and dry over anhydrous sodium sulfate.
AcOEt is distilled off and hardened with ether. AcOEt
- Reprecipitation with ether; 1.7 g (87%). Melting point 102~
103°C, [α] ²³ _D +20.3° (C = 0.33MeOH), Rf ¹ =
0.44 Elemental analysis C ₃₃ H ₃₈ O ₉ N ₄ Calculated value C62.45, H6.04, N8.83 Actual value C62.31, H6.28, N8.59 () Z−Tyr−(D)−Thr− Gly−Phe−NH
-Production of NH ₂ Z-Tyr-(D)-Thr-Gly-Phe-OCH ₃ 1.5g
(2.3 mmol) was dissolved in 30 ml of MeOH and cooled.
Add 0.5 ml of NH ₂ .NH ₂ .H ₂ O and leave at room temperature for 2 days. The precipitated crystals were collected by filtration and recrystallized from MeOH; 1.4 g (93%). Melting point 212~213℃,
[α] ²³ _D −25.9° (C = 0.32DMF), Rf ¹ = 0.12 Elemental analysis As C ₃₂ H ₃₈ O ₈ N ₆ Calculated value C60.55, H6.04, N13.24 Actual value C60.31, H6 .25, N12.98 () Z−Tyr−(D)−Thr−Gly−Phe−NH
-NH-CO-( _CH2 ) _{3 -} CH3 Production Z-Tyr-(D)-Thr-Gly-Phe-NH- _NH2
() of Example 5 using 540 mg and 0.14 ml of n-valeric acid.
The desired product was obtained in the same manner as; 420 mg (69%). Melting point 172-173°C, Rf ¹ =0.45. [α] ²⁴ _D −17.5゜(C=
0.39DMF). Elemental analysis C ₃₇ H ₄₆ O ₉ N ₆ Calculated value C61.82, H6.45, N11.69 Actual value C61.66, H6.44, N11.43 () H−Tyr−(D)−Thr−Gly −Phe−NH
-NH-CO-( _CH2 ) _{3- CH3} Production Z-Tyr-(D)-Thr-Gly-Phe-NH-NH
-CO-(CH ₂ ) ₃ -CH ₃ 250 mg () of Example 5
Deprotect and purify in the same manner as above to obtain 105 mg of the desired product. Rf ² =0.50, [α] ²⁴ _D +10.8゜(C=
0.31MeOH). Amino acid analysis: Thr0.97(1), Gly1.00(1), Tyr1.10
(1), Phe1.15(1) average recovery rate 78%. Example 17 H-Tyr-(D)-Ala-Gly-Phe-
Production of NH-NH-CO- _CH3 () Z-Tyr-(D)-Ala-Gly-Phe-
Production of NHNH-CO-CH ₃ Same as in Example 11 () using Z-Tyr-(D)-Ala-Gly-OH (0.66 g) and Z-Phe-NHNH-CO-CH ₃ (0.59 g). Synthesize as; 0.82g,
Melting point 210-211℃, [α] ^23D _-22.9゜(C=0.31,
DMF), Rf ¹ = 0.20 Elemental analysis C ₃₃ H ₃₈ O ₈ N ₆・H ₂ O Calculated value C59.63, H6.07, N12.64 Actual value C59.99, H6.28, N12.62 () H-Tyr-(D)-Ala-Gly-Phe-
Production of NHNH-COCH ₃ Z-Tyr-(D)-Ala-Gly-Phe-NHNH-
Using CO-CH ₃ (0.35 g), the same procedure as in Example 11 () is carried out to obtain 0.16 g of the desired product. [α]
²³ _D +21.6゜(C=0.30, MeOH), Rf ² =0.40, Amino acid analysis values: Gly1.00, Ala0.90, Tyr0.95, Phe0.97
(Average recovery rate 82%) Example 18 H-Tyr-(D)-Nva-Gly-Phe-
Production of NHNH-CO-CH ₃ (Nva=Norvaline) () Production of Z-Gly-Phe-NHNH-CO-CH ₃ Z-Phe-NHNH-CO-CH ₃ (42.6g)
Dissolve in MeOH (400ml) and perform catalytic reduction using palladium black as a catalyst. Filter the catalyst and MeOH
was distilled off and the residue was dissolved in DMF (100ml). Add Z-Gly-ONB (44 g) to this and stir at room temperature for 10 hours. DMF is distilled off, ether is added to the residue, and the powder is collected by filtration. Crystallizes from aqueous acetonitrile; 45.2g, melting point 154-155℃,
[α] ²³ _D −0.9° (C=0.50, DMF), Rf ¹ =0.52. Elemental analysis As C ₂₁ H ₂₄ O ₅ N ₄ Calculated value C61.15, H5.87, N13.59 Actual value C61, 09, H5.69, N13.18 () Z−(D)−Nva−Gly−Phe -NHNH-
Production of CO-CH ₃ Z-Gly-Phe-NHNH-CO-CH ₃ (5g)
Dissolve in MeOH (100ml) and perform catalytic reduction using palladium black as a catalyst. The catalyst is filtered off, MeOH is distilled off, and the precipitated crystals are collected by filtration. This crystal (H
−Gly−Phe−NHNH−CO−CH ₃ ) out of 3.5g
0.89g and Z-(D)-Nva-ONB(Z-(D)-Nva
-Synthesized from 0.80 g of OH and 0.63 g of HONB by the DCC method) was dissolved in DMF (10 ml) and stirred at room temperature for 10 hours. DMF was distilled off, ether was added to the residue, and the precipitate was collected by filtration. Crystallize from acetonitrile; 1.3 g. Melting point 235-237℃, [α] ²³ _D + 3.4゜ (C = 0.45, DMF), Rf ¹ = 0.44 Elemental analysis Calculated value as C ₂₆ H ₃₃ O ₆ N ₅ C61.04, H6.50, N13. 69 Actual value C61.33, H6.62, N13.66 () Z−Tyr−(D)−Nva−Gly−Phe−
Production of NHNH-CO- _CH3 Z-(D)-Nva-Gly-Phe-NHNH-CO-
CH ₃ (0.92 g) was dissolved in MeOH (50 ml) and catalytic reduction was performed using palladium black as a catalyst. The catalyst is filtered off, the MeOH is distilled off, and the residue is dissolved in DMF (10 ml). Add Z-Tyr-ONB (0.86 g) to this and stir at room temperature for 10 hours. DMF is distilled off, ether is added to the residue, and the precipitate is collected by filtration. Crystallize with acetonitrile; 0.85 g. melting point
210~211℃, [α] ²³ _D −18.3° (C=0.48, DMF),
^Rf1 =0.38. Elemental analysis C ₃₅ H ₄₂ O ₈ N ₆・1/2H ₂ O Calculated value C61.48, H6.33, N12.29 Actual value C61.39, H6.34, N12.09 () H−Tyr−( D) -Nva-Gly-Phe-
Production of NHNH-CO- _CH3 Z-Tyr-(D)-Nva-Gly-Phe-NHNH-
Dissolve CO-CH ₃ (0.40 g) in MeOH (50 ml) and perform catalytic reduction using palladium black as a catalyst. The catalyst was filtered off, MeOH was distilled off, and the residue was diluted with a small amount of 0.1N
- Dissolve in acetic acid water and apply to Sephadex LH-20 column (2.5 x 120 cm). Elute with 0.1N aqueous acetic acid, collect fractions from 320 ml to 335 ml, and freeze-dry to obtain 160 mg of the target product. [α] ²³ _D +22.6° (C = 0.30, MeOH), Rf ² = 0.44, amino acid analysis values: Gly 1.00, Nva 1.10, Tyr 0.99, Phe 1.07 (average recovery rate 83%). Example 19 H-Tyr-(D)-Ser-Gly-Phe-
Production of NHNH-CO- _CH3 (Ser=serine) () Z-(D)-Ser-Gly-Phe-NHNH-
Production of CO-CH ₃ H-Gly-Phe- obtained in () of Example 18
NHNH-CO- _CH3 (0.69g) and Z-(D)-Ser-
Using ONB (1.1 g), 1.2 g of the target product was obtained in the same manner as in Example 18 (). Melting point 168-169℃, [α] ²³ _D +3.4゜ (C=0.5, DMF),
^Rf1 =0.25. Elemental analysis C ₂₄ H ₂₉ O ₇ N As ₅ Drive value C57.72, H5.85, N14.02 Actual value C57.33, H5.95, N13.50 () Z−Tyr−(D)−Ser−Gly −Phe−
Production of NHNH-CO- _CH3 Z-(D)-Ser-Gly-Phe-NHNH-CO-
Using CH ₃ (0.75 g) and Z-Tyr-ONB (0.72 g), 0.86 g of the desired product was obtained in the same manner as in Example 18 (). Melting point 184-186℃, [α] ²³ _D -18.3゜(C=0.30,
DMF), Rf ¹ = 0.19. Elemental analysis C ₃₃ H ₃₈ O ₉ N ₆ Calculated value C59.81, H5.78, N12.68 Actual value C59.69, H5.82, N12.51 () H-Tyr-(D)-Ser-Gly −Phe−
Production of NHNH-CO- _CH3 Z-Tyr-(D)-Ser-Gly-Phe-NHNH-
Using CO-CH ₃ (0.40 g), 0.21 g of the target product was obtained in the same manner as in Example 18 (). [α] ²³ _D +11.4° (C = 0.31, MeOH), Rf ² = 0.21. Amino acid analysis values: Ser0.85, Gly1.00, Tyr0.90, Phe0.92. Example 20 H-Tyr-(D)-Thr-Gly-Phe-
Production of NHNH-CO- _CH3 () Z-(D)-Thr-Gly-Phe-NHNH-
Production of CO-CH ₃ H-Gly-Phe- obtained in () of Example 18
NHNH-CO- _CH3 (0.50g) and Z-(D)-Thr-
Using OH (0.48 g), 0.71 g of the desired product was obtained in the same manner as in Example 18 (). Melting point 193-195℃, [α]
²⁵ _D +5.3゜ (C = 0.41, DMF), Rf ¹ = 0.41 Elemental analysis C ₂₅ H ₃₁ O ₇ N ₅ Calculated value C58.47, H6.08, N13.64 Actual value C58, 13, H6. 13, N13.25 () Z−Tyr−(D)−Thr−Gly−Phe−
Production of NHNH-CO- _CH3 Z-(D)-Thr-Gly-Phe-NHNH-CO-
0.51 g of the target product was obtained in the same manner as in Example 18 () using CH ₃ (0.46 g) and Z-Tyr-ONB (0.43 g). Melting point 208-209℃, [α] ^25D _-19.0゜(C=0.50
，
DMF), Rf ¹ = 0.28. Elemental analysis C ₃₄ H ₄₀ O ₉ N ₆・1/2H ₂ O Calculated value C59.55, H6.03, N12.25 Actual value C59.79, H6.11, N12.40 () H−Tyr−( D) -Thr-Gly-Phe-
Production of NHNH-CO- _CH3 Z-Tyr-(D)-Thr-Gly-Phe-NHNH-
Using CO-CH ₃ (0.30 g), 0.11 g of the desired product was obtained in the same manner as in Example 18 (). [α] ²⁵ _D +12.0° (C = 0.26, MeOH), Rf ² = 0.24, amino acid analysis values: Thr1.02, Gly1.00, Tyr0.98, Phe1.04 Example 21 H-Tyr-(D ) −Met−Gly−Phe−
Production of NHNH-CO-CH ₂ -CH ₃ (Met = methionion) () Production of Z-Phe-NHNH-CO-CH ₂ CH ₃ Z-Phe-NHNH ₂ (12.5g) and propionic anhydride (5.8ml) 9.8 g of the target product was obtained in the same manner as in Example 4 (). Melting point 203-204℃, [α]
²¹ _D −17.6゜(C=0.46, DMF), Rf ¹ =0.60 Elemental analysis As C ₂₀ H ₂₃ O ₄ N ₃ Calculated value C65.92, H6.27, N10.25 Actual value C65.87, H6.72 , N9.93 () Z−Gly−Phe−NHNH−CO−
Production of CH ₂ CH ₃ Using Z-Phe-NHNH-CO-CH ₂ CH ₃ (2.2 g) and Z-Gly-ONB (2.2 g), 1.8 g of the target product was prepared in the same manner as in () of Example 18. get. melting point
151−152°C, [α] ²¹ _D −1.2° (C=0.50, DMF),
^Rf1 =0.46. Elemental analysis As C ₂₂ H ₂₆ O ₅ N ₄ Calculated value C61.96, H6.15, N13.14 Actual value C62.25, H6.23, N12.85 () BOC−(D)−Met−Gly−Phe −NHNH
Production of −CO−CH ₂ −CH ₃ Z−Gly−Phe−NHNH−CO−CH ₂ CH ₃ (0.77
g) and BOC-(D)-Met-OH (0.45g),
0.75 g of the target product was obtained in the same manner as in Example 18 (). Melting point 172-173℃, [α] ²⁴ _D +6.9゜(C=0.33,
DMF), Rf ¹ = 0.50. Elemental analysis C ₂₄ H ₃₇ O ₆ N ₅ S Calculated value C55.04, H7.12, N13.37, S6.12 Actual value C55.05, H6.94, N13.52, S6.02 () BOC−Tyr -(D)-Met-Gly-Phe-
Synthesis of NHNH-CO-CH ₂ CH ₃ BOC-(D)-Met-Gly-Phe-NHNH-CO
-CH ₂ -CH ₃ (0.50 g) is dissolved in TFA (5 ml) and left at room temperature for 20 minutes. TFA is distilled off, ether is added to the residue, and the precipitate is filtered and dried. This powder was dissolved in DMF (10 ml), cooled, and TEA (0.14
ml) and BOC-Tyr-ONB (0.44 g) and stir at room temperature overnight. Distill the DMF and remove the residue.
Extract with AcOEt (100 ml), wash with water, and dry with anhydrous sodium sulfate. AcOEt is distilled off and the precipitated gel-like substance is collected by filtration. Recrystallize from AcOEt;
0.45g. Melting point 159-160℃, [α] ²⁴ _D +0.9゜(C=0.46
，
DMF), Rf ¹ = 0.29. Elemental analysis C ₃₃ H ₄₆ O ₈ N ₆ As S Calculated value C57.70, H6.75, N12.23, S4.67 Actual value C57.92, H6.85, N11.99, S4.49 () H- Tyr-(D)-Met-Gly-Phe-
Production of NHNH-CO-CH ₂ CH ₃ BOC-Tyr-(D)-Met-Gly-Phe-NHNH
-CO-CH ₂ CH ₃ (0.40 g) is dissolved in TFA (4 ml) and left at room temperature for 20 minutes. TFA is distilled off, ether is added to the residue, and the precipitated gel-like substance is collected by filtration and dried. Dissolve this powder in 50 ml of water and use a column of Amberlite IRA-410 (acetic acid type) (2 x 5
cm). The effluent and washing liquid are combined and freeze-dried. Dissolve the obtained powder in a small amount of 0.1N acetic acid water,
Sephadex LH-20 column (2.5 x 120cm)
Put it on. Elute with 0.1N-acetic acid water, from 315ml
0.25 when fractions up to 330ml are collected and freeze-dried.
Obtain the object g. [α] ²¹ _D +20.0° (C=0.20,
MeOH), Rf ² = 0.46 Amino acid analysis values: Gly1.00, Met0.77, Tyr0.80,
Phe1.02 Example 22 H-Tyr-(D)-Phe-Gly-Phe-
Production of NHNH-CO- _CH3 () BOC-(D)-Phe-Gly-Phe-NHNH
-Production of COCH ₃ H-Gly-Phe-NHNH-CO-CH ₃ (0.44g)
and BOC-(D)-Phe-ONB (0.68g) in the same manner as in () of Example 18 to obtain the target product (0.68g).
get. Melting point 208-209℃, [α] ²³ _D +3.3゜(C=
0.45, DMF), Rf ¹ = 0.51 Elemental analysis As C ₂₇ H ₃₅ O ₆ N ₅ Calculated value C61.70, H6.71, N13.33 Actual value C61.45, H6.65, N13.46 () Z- Tyr-(D)-Phe-Gly-Phe-
Production of NHNH-CO- _CH3 BOC-(D)-Phe-Gly-Phe-NHNH-CO
-CH ₃ (0.51 g) and Z-Tyr-ONB (0.46 g) were used to prepare the target product (0.51
g) is obtained. Melting point 203-204℃, [α] ^23D _-13.0゜(C
= 0.44, DMF), Rf ¹ = 0.38 Elemental analysis C ₃₉ H ₄₂ O ₈ N ₆ Calculated value C64.80, H5.86, N11.63 Actual value C64.70, H5.95, N11.40 () H -Tyr-(D)-Phe-Gly-Phe-
Production of NHNH-CO- _CH3 Z-Tyr-(D)-Phe-Gly-Phe-NHNH-
CO-CH ₃ (0.45 g) was dissolved in MeOH (50 ml) and catalytic reduction was performed using palladium black as a catalyst. The catalyst is filtered off and MeOH is distilled off. Add ether to the residue to make a powder, and filter it. Crystallize from acetonitrile; 0.22 g. [α] ²³ _D −11.4゜(C=0.35
，
MeOH), Rf ² = 0.37. Amino acid analysis: Gly 1.00, Tyr 0.98, Phe 1.95.

Claims (1)

[Claims] 1. General formula [In the formula, R ₁ represents hydrogen. R ₂ represents the side chain of D-α-amino acid. R ₃ represents hydrogen or alkyl having 1 to 2 carbon atoms. R ₄ is hydrogen, a linear or branched alkyl having 1 to 8 carbon atoms substituted with a hydroxyl group, a hydroxyl group, an amino group,
It represents a linear or branched saturated aliphatic acyl having 1 to 8 carbon atoms which may be substituted with alkoxy having 1 to 4 carbon atoms or halogen. ] A tetrapeptide derivative having an analgesic effect and a pharmacologically acceptable acid addition salt thereof. 2. The tetrapeptide derivative and pharmacologically acceptable acid addition salt thereof according to claim 1, wherein R ₄ is a linear or branched aliphatic acyl having 3 to 6 carbon atoms. 3 General formula [In the formula, R ₁ represents hydrogen. _R'2 represents a side chain of D-α-amino acid which may have a protecting group. R ₃ represents hydrogen or alkyl having 1 to 2 carbon atoms. _R'4 is hydrogen, a linear or branched alkyl group having 1 to 8 carbon atoms substituted with a hydroxyl group, or a hydroxyl group, an amino group, or a 1 to 4 carbon group;
A linear or branched chain having 1 to 8 carbon atoms which may be substituted with alkoxy or halogen.
represents a saturated aliphatic acyl which may have a protecting group. Y ₁ represents a protecting group. Y ₂
and Y ₃ each represent hydrogen or a protecting group. ] A general formula characterized by subjecting a compound represented by to a protecting group elimination reaction [In the formula, R ₁ and R ₃ have the same meanings as above.
R ₂ represents the side chain of D-α-amino acid. R ₄ is hydrogen, a linear or branched alkyl having 1 to 8 carbon atoms substituted with a hydroxyl group, or a straight chain optionally substituted with a hydroxyl group, an amino group, an alkoxy having 1 to 4 carbon atoms, or a halogen represents a saturated or branched aliphatic acyl having 1 to 8 carbon atoms. ] A method for producing a tetrapeptide derivative having analgesic effect.