JPS6257639B2 - - 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 was isolated and its chemical structure determined, but although these substances exhibit morphan-like effects when administered directly into the brain, they do not have analgesic effects when administered intravenously. Not shown. On the other hand, β
- β-endorphin, which is considered to be derived from the carboxy terminus of 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 synthesis It is extremely difficult to manufacture and supply in large quantities as a pharmaceutical product. 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. The present invention provides (1) General formula () [In the formula, R ₁ and R ₂ each represent hydrogen or methyl, and R ₃ represents hydrogen, or lower aliphatic acyl optionally substituted with a hydroxyl group, lower alkoxy, lower alkylthio, lower alkylthiooxide, halogen, or oxo group. , respectively. ]
A tetrapeptide derivative represented by () and a pharmacologically acceptable acid addition salt thereof. and (2) general formula () [In the formula, R ₁ and R ₂ each represent hydrogen or methyl, and R ₃ ' represents hydrogen, or a lower aliphatic acyl which may be substituted with a hydroxyl group, lower alkoxy, lower alkylthio, lower alkylthiooxide, halogen or oxo group. and may have a protecting group, Y ₁ represents a protecting group, and Y ₂ and Y ₃ represent hydrogen or a protecting group, respectively. ] General formula () characterized by subjecting the compound () represented by ] to a protecting group elimination reaction [In the formula, R ₁ and R ₂ have the same meanings as above. _R3
represents hydrogen, or lower aliphatic acyl optionally substituted with a hydroxyl group, lower alkoxy, lower alkylthio, lower alkylthiooxide, halogen or oxo group. This is a method for producing a tetrapeptide derivative () represented by 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 Met(O): Methionine sulfoxide Mephe: N-methyl-phenylalanine Tyr: Tyr: Tyrosine MeTyr: N-methyl-tyrosine Also, compounds commonly used in the text will be displayed using the following abbreviations. 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 Ester TEA: Triethylamine THF: Tetrahydrofuran TFA: Trifluoroacetic acid When an amino acid or its residue is expressed in the above abbreviation herein, it means the L-form unless otherwise specified, and for the D-form, the (D )-. As the lower aliphatic acyl represented by R ₃ ,
Those having 1 to 8 carbon atoms are preferred, and examples thereof include formyl, acetyl, propionyl, isopropionyl, n-butyryl, isobutyryl, valeryl, isovaleryl, n-caproyl, isocaproyl, n-heptanoyl, etc. Particularly preferred are those having 2 to 5 carbon atoms. The lower alkoxy which may be substituted with the above group is preferably one having 1 to 4 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy. Among them, methoxy, ethoxy, and n-propoxy are particularly preferred. The lower alkylthio optionally substituted with the above acyl preferably has 1 to 4 carbon atoms, and examples include methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, isobutylthio, tert- Examples include butylthio, among others methylthio,
Particularly preferred are ethylthio, n- or iso-butylthio, and the like. The lower alkyl thioxide which may be substituted with the above acyl preferably has 1 to 4 carbon atoms, and examples thereof include methyl thioxide, ethyl thioxide, n-propyl thioxide, isopropyl thioxide, n-propyl thioxide,
-butyl thioxide, isobutyl thioxide, tert-butyl thioxide, etc.
Among these, methylthiooxide, ethylthiooxide, n- or iso-butylthiooxide, etc. are particularly preferred. Examples of the halogen that may be substituted with the above acyl include chlorine, fluorine, and bromine. In the above general formula, examples of the protecting group represented by Y ₁ include benzyloxycarbonyl, t-butoxycarbonyl, t-amyloxycarbonyl, isobornyloxycarbonyl, adamantyloxycarbonyl, chloro- or nitro-substituted benzyloxycarbonyl , o-phenylthio, diphenylphosphinothioyl, and the like. Examples of the protecting group represented by Y ₂ include benzyl and t-butyl. Regarding the protecting group for R ₃ ', when protecting the hydroxyl group of each basic group, examples of the protecting group include those defined as Y ₂ 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 interest () 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 M.
A. Ondetti, Peptide Synthesis
Synthesis), Inter science, New York, 1966
Year; The Proteins by FMFinn and K. Hofmann, Volume 2, H. Nenrath, RL
Edited by Hi11, Academic Press Inc. New York,
1976; “Peptide Synthesis” by Nobuo Izumiya et al. Maruzen Co., Ltd.
Methods described in 1975, such as the azide method, chloride method, acid anhydride method, mixed acid anhydride method,
DCC method, active ester method, Woodward reagent K
, the carbodiimidazole method, the redox method, and the DCC/HONB method. In some cases, the NCA method (N-carboxyanhydride; a method that uses 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
It can also be protected in the form of esters such as -butyl, t-amyl, etc. As the 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, such as benzyl, tosyl, 2,4-
Examples include dinitrophenyl, t-butyloxycarbonyl, carbobenzoxy and the like. Examples of protecting groups for the hydroxyl group of tyrosine include benzyl,
Examples include ethers such as t-butyl. Raw materials with activated carboxyl groups include, for example, corresponding acid anhydrides, azides, active esters [alcohols (e.g., pentachlorophenol, 2,4,5-trichlorophenol, 2,
4-dinitrophenol, cyanomethyl alcohol, p-nitrophenol, N-hydroxy-
5-norbornene-2,3-dicarboximide, N-hydroxysuccinimide, N-hydroxyphthalimide, N-hydroxybenztriazole], and the like. As the 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 carboxyl and amino groups of A and B in the above-mentioned states 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 water-containing 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】

[Table] The desorption conditions shown in Table 1 are, for example,
Catalytic reduction using palladium black, palladium on carbon, platinum, etc. as a catalyst, acid decomposition using trifluoroacetic acid, dilute hydrochloric acid, methanesulfonic acid, etc. In addition to these, 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, partitioning, reprecipitation, recrystallization, column chromatography, etc. The compound () of the present invention can also be produced using sulfoxidized methionine, which is one of the raw material compounds, or can be produced using the general formula () [In the formula, R ₁ , R ₂ and R ₃ have the same meanings as above. A tetrapeptide derivative represented by the general formula () can also be produced by oxidizing an intermediate of the compound () represented by the formula () and then subjecting the compound () to an oxidation reaction. For the above-mentioned oxidation reaction, for example, hydrogen peroxide, sodium perchlorate, sodium perborate, chloramine T, tribromocresol, N-chlorosuccimide, and chloroauric acid are used as oxidizing agents. The amount of the oxidizing agent is about 1 equivalent or more, preferably about 1 to 10 equivalents, based on the compound (). Water, acetic acid, alcohol, or a mixture thereof is used as a solvent, and the reaction is carried out at an appropriate temperature of -20°C to 40°C. The peptide derivative () obtained by oxidizing the compound () in this manner is collected by peptide separation means such as partitioning, reprecipitation, recrystallization, and column chromatography after completion of the reaction. Peptide () can be obtained by forming a salt with an organic acid, an inorganic acid, etc. by a conventional method, but the acids generally include acetate, citrate, tartrate, hydrochloride, and sulfate. etc. are preferable. 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 18-22 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 showed 1 in the above test.
Analgesic effects with a prolongation rate of 40% or more with a peak duration of 5 to 60 minutes are observed at doses of mg/Kg or less, and in the case of particularly effective compounds, clear analgesic effects are observed even at minute doses of 0.1 mg/Kg or less. The compound () of the present invention can be administered subcutaneously by the hot plate method using mice as described above.
It shows a clear analgesic effect at a dose in the range of 0.05 to 1 mg/Kg, and is more effective than β-endorphin, so it is useful as an analgesic.
It is also expected to have neuroactive effects. Furthermore, the compound () of the present invention exhibits a strong gastrointestinal motility inhibiting effect, and is therefore useful as a therapeutic agent for gastrointestinal disorders such as diarrhea. Therefore, the compound () of the present invention and its pharmacologically acceptable acid addition salts can be used to treat terminal cancer and severe pain in mammals (e.g., mice, rats, rabbits, dogs, monkeys, humans, etc.). It can be used as an analgesic and as a treatment for gastrointestinal disorders such as diarrhea. It is also useful for mental illnesses such as schizophrenia. 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 0.01 mg to 20 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 0.01mg to 20mg. 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. In the case of intravenous or subcutaneous injection, it is preferable to use, for example, a compound () dissolved in physiological saline at a concentration of 1 to 20 mg/ml. Further, Compound () and certain compounds of () have analgesic effects similar to Compound (). In the compounds (), () and () of _the present invention, _{R 3}
Alternatively, a lower aliphatic acyl compound in which R ₃ ' may have a substituent may have a stronger effect. 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 Kieselgel 60F-254 (Merck & Co., Germany). 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 1 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-CH ₂ -CH ₃ () Production of Z-Phe-NH-NH ₂ Dissolve 59 g of Z-Phe-OH in 200 ml of MeOH,
Add 5 ml of 6N-hydrochloric acid-dioxane to this and leave it at room temperature overnight. MeOH is distilled off, and the residue is dissolved in 300 ml of diethyl ether and washed with water. After drying over anhydrous sodium sulfate, diethyl ether is distilled off and the residue is dissolved in 150 ml of MeOH. Add 16 ml of hydrazine hydrate to this and leave it at room temperature for 2 days.
MeOH is distilled off and the precipitated crystals are filtered using petroleum benzine. After washing thoroughly with diethyl ether
AcOEt - Recrystallize with petroleum benzine. Yield 49
g (79%), melting point 159-160°C, Rf ¹ = 0.62,
[α] ²¹ _D -9.8° (c=0.8, DMF) Elemental analysis As C ₁₇ H ₁₉ O ₃ N ₃ Calculated values C 65.16, H 6.11, N 13.41 Actual values C 65.02, H 6.32, N 13.20. () Production of Z-Phe-NH-NH-CO-CH ₂ -CH ₃ Dissolve 12.5 g of Z-Phe-NH-NH ₂ in 30 ml of DMF, cool, add 5.8 ml of propionic anhydride and 3 ml of pyridine, and stir overnight at room temperature. Stir. Add water to the reaction solution and filter the precipitate. Recrystallize from acetonitrile. Yield 9.8g (68
%), melting point 203-204°C, Rf ¹ =0.60, [α] ²¹ _D −
17.6° (c=0.46, DMF) Elemental analysis As C ₂₀ H ₂₃ O ₄ N ₃ Calculated values C 65.92, H 6.27, N 10.25 Actual values C 65.87, H 6.72, N 9.93. () Z−Gly−Phe−NH−NH−CO−CH ₂ −
Production of CH ₃ Z-Phe-NH-NH-CO-CH ₂ -CH ₃ 2.2g
Dissolve in 50ml of MeOH 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 10 ml of DMF. Z- to this
Add 2.2 g of Gly-ONB and stir overnight at room temperature. DMF is distilled off, and the residue is extracted with 100 ml of AcOEt, washed with water, and dried over anhydrous sodium sulfate.
AcOEt is distilled off and the precipitated crystals are collected by filtration.
AcOEt - Recrystallize with petroleum benzine. Yield 1.8
g (71%), melting point 151-152°C, Rf ¹ = 0.46,
[α] ²¹ _D −1.2° (c=0.5, DMF), elemental analysis as C ₂₂ H ₂₆ O ₅ N ₄ Calculated values: C 61.96, H 6.15, N 13.14 Actual values: C 62.25, H 6.23, N 12.85. () BOC−(D)−Met(O)−Gly−Phe−NH−
Production of NH−CO−CH ₂ −CH ₃ Z−Gly−Phe−NH−NH−CO−CH ₂ −
Dissolve 1.0 g of CH ₃ in 50 ml of MeOH and perform catalytic reduction using palladium black as a catalyst. Filter out the catalyst
Distill MeOH and dissolve the residue in 20 ml of acetonitrile. This is added to BOC-D-Met-ONB (BOC
-D-Met-OH600mg and HONB470mg DCC
(prepared by method) and stir overnight at room temperature. Add 0.5ml of 30% hydrogen peroxide solution to the reaction solution and incubate at room temperature for 10 minutes.
Stir the time. Destroy excess hydrogen peroxide with palladium black and distill off acetonitrile. The residue was extracted with 100 ml of n-BuOH and washed with water. n-
BuOH is distilled off and solidified with diethyl ether. Yield 1.1g (88%), melting point 121-122℃, Rf ¹
= 0.40, [α] ²¹ _D 0° (c = 0.5, DMF), elemental analysis Calculated value as C ₂₄ H ₃₇ O ₆ N ₅ S・1/2H ₂ O
C 52.53, H 6.97, N 12.76, S 5.84 Actual value
C 52.41, H 6.99, N 12.46, S 5.61. () BOC−Tyr−(D)−Met(O)−Gly−Phe−
Production of NH-NH-CO-CH ₂ -CH ₃ BOC-(D)-Met(O)-Gly-Phe-NH-NH
Dissolve 0.82 g of -CO-CH ₂ -CH ₃ in 10 ml of TFA and leave at room temperature for 20 minutes. TFA is distilled off and the residue is filtered with diethyl ether. Dissolve this powder in 10ml of DMF and
Add 710 mg of ONB and 0.3 ml of TEA and stir overnight at room temperature. DMF is distilled off and the residue is n-
Extract with 100ml of BuOH and wash with water. n-BuOH was distilled off and solidified with diethyl ether. Yield 0.75g
(72%), melting point 134-135℃, Rf ¹ = 0.20, [α] ²¹ _D
0° (c=0.5, DMF), elemental analysis Calculated value as C ₃₃ H ₄₆ O ₉ N ₆ S・H ₂ O
C 54.98, H 6.71, N 11.66, S 4.44 Actual value
C 54.72, H 6.82, N 11.16, S 4.43. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO- _CH2 - _CH3 production BOC-Tyr-(D)-Met(O)-Gly-Phe-NH
Dissolve 400 mg of -NH-CO-CH ₂ -CH ₃ in 4 ml of TFA and leave at room temperature for 20 minutes. TFA is distilled off and the residue is filtered with diethyl ether. Dissolve this powder in 30ml of water and use Amberlite IRA.
-410 (acetic acid form, 2 x 6 cm) column,
The effluent and washing liquid are combined and freeze-dried. This powder is dissolved in a small amount of 0.1N aqueous acetic acid, passed through a Sephadex LH-20 column, and then eluted with aqueous 0.1N acetic acid. Collect the elution fractions from 310 ml to 330 ml and lyophilize. Yield 310 mg, Rf ² =
0.35, [α] ²¹ _D +24.3° (c = 0.35, MeOH), amino acid analysis (hydrochloric acid hydrolyzate): Gly1.00,
Met0.57, Tyr0.88, Phe1.00. Example 2 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-CH ₂ -CH ₂ -CH ₃ () Z-Tyr-(D)-Met(O)-Gly-Phe-OEt
Production of H-Phe-OEt, hydrochloride (4.8 g) was dissolved in 50 ml of acetonitrile, cooled, and TEA 2.8 ml and Z-Gly
- Add 7.4 g of ONB and stir overnight at room temperature. The solvent was distilled off and the residue was extracted with 200ml of AcOEt.
Wash with water and dry with anhydrous sodium sulfate. AcOEt is distilled off, the residue is dissolved in 100 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 50 ml of acetonitrile. To this BOC−(D)
−Met−ONB [BOC−(D)−Met−OH4.7g and
Prepared from 3.7g of HONB using the DCC method. ] and stir overnight at room temperature. Add 1 ml of N-hydrochloric acid to the reaction solution.
Add 2.2 ml of 30% hydrogen peroxide solution and stir at room temperature for 10 hours. Excess hydrogen peroxide is crushed with palladium black and acetonitrile is distilled off.
Extract with 200 ml of AcOEt, wash with water, and dry with anhydrous sodium sulfate. Distill AcOEt and remove the residue
Dissolve in 30ml of TFA and leave at room temperature for 20 minutes.
TFA is distilled off and the residue is filtered with diethyl ether. Add this powder to acetonitrile 60
ml, cool and add 2.5ml of TEA and Z-Tyr-
Add 7.2 g of ONB and stir overnight at room temperature. The solvent was distilled off, and the residue was extracted with 200 ml of AcOEt, washed with water, and dried over anhydrous sodium sulfate.
When AcOEt is distilled off, crystals will precipitate and should be collected by filtration. Recrystallize from AcOEt. Yield 6.9 g, melting point 162-163°C, Rf ¹ = 0.42, [α] ²¹ _D -3.2° (c
= 0.37, DMF), calculated value as elemental analysis C ₃₅ H ₄₂ O ₉ N ₄ S
C 60.50, H 6.09, N 8.06, S 4.61 Actual value
C 60.45, H 6.38, N 8.07, S 4.37. () Z−Tyr−(D)−Met(O)−Gly−Phe−NH
-Production of NH ₂ Z-Tyr-(D)-Met(O)-Gly-Phe-
6.0g of OEt in 20ml of ethyl alcohol - 20ml of DMF
Dissolve the solution in a mixed solvent, add 2 ml of hydrazine hydrate, and let stand at room temperature for 2 days. Ethyl alcohol is distilled off, diethyl ether is added, and the precipitated crystals are collected by filtration. Boil and wash with acetonitrile.
Yield 5.0g, melting point 182-184℃, Rf ¹ = 0.10,
[α] ²¹ _D −13.4° (c=0.44, DMF), elemental analysis Calculated value as C ₃₃ H ₄₀ O ₈ N ₆ S
C 58.21, H 5.92, N 12.34, S 4.71 Actual value
C 57.95, H 5.98, N 12.10, S 4.55. () Z−Tyr−(D)−Met(O)−Gly−Phe−NH
Production of -NH-CO-CH ₂ -CH ₂ -CH ₃ Z-Tyr-(D)-Met(O)-Gly-Phe-
Dissolve 0.51 g of NHNH ₂ and 0.08 ml of butyric acid in 5 ml of DMF, cool to 0°C, add 120 mg of HOBT and 190 mg of DCC, and stir at 0°C for 5 hours at room temperature overnight. DMF
was distilled off and the residue was solidified with diethyl ether. Reprecipitate with MeOH-ether. Yield 410
mg, melting point 161-163°C, Rf ¹ =0.31, [α] ²¹ _D -8.8
° (c=0.34, DMF), elemental analysis Calculated value as C ₃₇ H ₄₆ O ₉ N ₆ S
C 59.18, H 6.17, N 11.19, S 4.27 Actual value
C 58.85, H 6.02, N 10.86, S 4.05. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
Production of -NH-CO-CH ₂ -CH ₂ -CH ₃ Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
300 mg of NH--CO--CH ₂ --CH ₂ --CH ₃ is dissolved in 3 ml of methanesulfonic acid in the presence of 0.3 ml of anisole and left at room temperature for 30 minutes. Add diethyl ether to the reaction mixture and let stand at -20°C for 30 minutes. Remove the supernatant, dissolve the oil in 30 ml of water, and pass through a column of Amberlite IRA-410 (acetic acid form, 2 x 6 cm). The effluent and washing liquid are combined and freeze-dried.
The obtained powder was dissolved in a small amount of 0.1N aqueous acetic acid and applied to a Sephadex LH-20 column (2.5 x 125
cm). Elute with 0.1N acetic acid water, 275~
Collect up to 290 ml of eluate fractions and lyophilize. Yield 110 mg, Rf ² = 0.37, [α] ²¹ _D +23.7° (c = 0.32, MeOH), amino acid analysis (hydrochloric acid hydrolyzate): Gly 1.00, Met 0.38, Tyr 0.91,
Phe1.05. Example 3 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-CH( _CH3 ) ₂ () Z-Tyr-(D)-Met(O)-Gly-Phe-NH
Production of -NH-CO-CH( _CH3 ) ₂ Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
Synthesis is carried out in the same manner as in Example 2 () using 0.51 g of NH ₂ and 0.08 ml of isobutyric acid. Yield 440 mg, melting point 181-183°C, [α] ²¹ _D -8.3° (c = 0.37,
DMF), Rf ¹ = 0.32, elemental analysis Calculated value as C ₃₇ H ₄₆ O ₉ N ₆ S
C 59.18, H 6.17, N 11.19, S 4.27 Actual value
C 58.77, H 6.01, N 10.82, S 4.15. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
Production of -NH-CO-CH( _CH3 ) ₂ Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
The desired product was obtained in the same manner as in Example 2 () using 300 mg of NH-CO-CH( _CH3 ) ₂ . Yield 105
mg, Rf ² = 0.37, [α] ²¹ _D +26.8° (c = 0.32,
MeOH), amino acid analysis (hydrochloric acid hydrolyzate):
Gly1.00, Met0.38, Tyr0.89, Phe1.02. Example 4 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO- _CH3 () Z-Tyr-(D)-Met(O)-Gly-Phe-NH
Using 0.51 g of -NH ₂ and 0.06 ml of acetic acid, 405 mg of the desired product was obtained in the same manner as in Example 2 (). melting point
180-182℃, Rf ¹ = 0.28, [α] ²¹ _D -9.1゜ (c =
0.30, DMF), elemental analysis Calculated value as C ₃₅ H ₄₂ O ₉ N ₆ S・H ₂ O
C 56.75, H 5.72, N 11.34, S 4.33 Actual value
C 56.48, H 5.62, N 11.08, S 4.05 () H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO- _CH3 production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
() of Example 2 using 300 mg of NH-CO-CH ₃
Obtain 130 mg of the target product in the same manner as above. ^Rf2 =
0.30, [α] ²¹ _D +19.6° (c = 0.27, MeOH), amino acid analysis (acid hydrolyzate): Gly1.00, Met0.35,
Tyr0.91, Phe1.02. Example 5 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-(CH ₂ ) ₃ CH ₃ () Z-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO-( _CH2 ) _{3 -} CH3 Production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
Using 0.47 g of NH ₂ and 0.1 ml of n-valeric acid, 0.39 g of the desired product was obtained in the same manner as in Example 2 (). Melting point 159-161℃, Rf ¹ = 0.33, [α] ^21D _-8.6゜(c
= 0.4, DMF), calculated value as elemental analysis C ₃₈ H ₄₈ O ₉ N ₆ S
C 59.66, H 6.32, N 10.98, S 4.19 Actual value
C 59.39, H 6.42, N 10.69, S 4.06. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO-( _CH2 ) _{3 -} CH3 Production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
Example 2 using 300 mg of NH-CO-( _CH2 ) _{3 -} CH3
Obtain 110 mg of the target product in the same manner as in ().
Rf ² = 0.38, [α] ²¹ _D +24.8° (c = 0.32,
MeOH), amino acid analysis (acid hydrolyzate):
Gly1.00, Met0.29, Tyr0.89, Phe1.00. Example 6 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-( _CH2 ) ₄ - _CH3 () Z-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO-( _CH2 ) _{4- CH3} Production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
430 mg of the desired product was obtained in the same manner as in Example 2 () using 0.51 g of NH ₂ and 0.09 ml of n-caproic acid. Melting point 170-172℃, Rf ¹ =0.34, [α] ^21D _-9.1
° (c=0.48, DMF), elemental analysis Calculated value as C ₃₉ H ₅₀ O ₉ N ₆ S
C 60.13, H 6.47, N 10.79, S 4.11 Actual value
C 59.88, H 6.42, N 10.53, S 4.22. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO-( _CH2 ) _{4- CH3} Production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
Example 2 using 350 mg of NH-CO-( _CH2 ) ₄ - _CH3
Obtain 210 mg of the target product in the same manner as in ().
Rf ² = 0.42, [α] ²¹ _D +25.7° (c = 0.30,
MeOH), amino acid analysis (hydrochloric acid hydrolyzate):
Gly1.00, Met0.29, Tyr0.88, Phe0.99. Example 7 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-( _CH2 ) ₂ -O- _CH2 - _CH3 () Z-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO-( _CH2 ) ₂ -O- _CH2 - _CH3 Production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
0.51 g of NH ₂ and 0.09 ml of β-ethoxypropionic acid
The target object is obtained in the same manner as in () of Example 2 using
Get 440mg. Melting point 159-161℃, Rf ¹ = 0.32,
[α] ²¹ _D -7.5゜ (c=0.48, DMF) Elemental analysis Calculated value as C ₃₈ H ₄₈ O ₁₀ N ₆ S
C 58.44, H 6.19, N 10.76, S 4.10 Actual values
C 58.22, H 6.31, N 10.59, S 3.99. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO-( _CH2 ) ₂ -O- _CH2 - _CH3 Production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
The target product 105 was prepared in the same manner as in Example 2 () using 350 mg of NH-CO-( _CH2 ) ₂ -O- _CH2 - _CH3 .
Get mg. Rf ² = 0.40, [α] ²¹ _D +25.1° (c =
0.37), Amino acid analysis (acid hydrolyzate): Gly1.00,
Met0.32, Tyr0.89, Phe0.98. Example 8 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-O-CH ₂ -CH ₃ () Production of BOC-(D)-Met-Gly-OEt 10.0g of BOC-(D)-Met-OH and 7.9g of HONB
Dissolve in THF100ml and cool to 0℃, DCC9.0g
Add and stir at 0℃ for 6 hours. Insoluble matter was filtered out, and 5.9 g of H-Gly-OEt hydrochloride was added to the filtrate.
Add 5.6ml of TEA and stir the solution at room temperature. THF
is distilled off, and the residue is extracted with 100 ml of AcOEt, washed with water, and dried over anhydrous sodium sulfate. AcOEt is distilled off and crystallized with petroleum benzene. AcOEt - Recrystallize with petroleum benzine. Yield: 9.8 g, melting point: 55-56°C, Rf ¹ = 0.70, [α] ²¹ _D +
12.5゜ (c=0.67, DMF), calculated value as elemental analysis C ₁₄ H ₂₆ O ₅ N ₂ S
C 50.28, H 7.83, N 8.37, S 9.59 Actual value
C 50.88, H 7.99, N 8.46, S 9.42. () Production of BOC-Tyr-(D)-Met-Gly-OEt 3.0g of BOC-(D)-Met-Gly-OEt and 15ml of TFA
Dissolve in and leave at room temperature for 10 minutes. TFA is distilled off and the residue is filtered with diethyl ether. Dissolve this powder in 20ml of THF and cool it.
Add 1.6 ml of TEA and 4.0 g of BOC-Tyr-ONB and stir at room temperature overnight. Distill the THF and remove the residue
Extract with 100 ml of AcOEt, wash with water, and dry with anhydrous sodium sulfate. AcOEt is distilled off and crystallized with petroleum benzene. Recrystallize from AcOEt.
Yield 3.1g, melting point 121-122℃, Rf ¹ = 0.62,
[α] ²¹ _D +16.8° (c=0.40, DMF), elemental analysis Calculated value as C ₂₃ H ₃₅ O ₇ N ₃ S
C 55.51, H 7.09, N 8.44, S 6.44 Actual value
C 55.32, H 6.93, N 8.25, S 6.27. () Production of BOC-Tyr-(D)-Met-Gly-OH 2.8g of BOC-Tyr-(D)-Met-Gly-OEt
Dissolve in 30 ml of MeOH, cool, add 12 ml of N-caustic soda water, and stir at room temperature for 1 hour. Cool, add 60 ml of 0.2N hydrochloric acid, collect the precipitated crystals by filtration, and wash with cold water. Yield 2.3g. Melting point 184−186
°C, Rf ¹ = 0.24, [α] ²¹ _D +13.3° (c = 0.46,
DMF), elemental analysis Calculated value as C ₂₁ H ₃₁ O ₇ N ₃ S
C 53.71, H 6.65, N 8.94, S 6.83 Actual value
C 54.32, H 6.71, N 8.49, S 6.70. () Production of BOC-Tyr-(D)-Met(O)-Gly-OH 2.2g of BOC-Tyr-(D)-Met-Gly-OH
Dissolve in a mixed solvent of ml of MeOH and 5 ml of acetic acid, add 0.6 ml of 30% hydrogen peroxide, and stir at room temperature for 5 hours. Add a small amount of palladium black and stir. When the catalyst is filtered off and the solvent is distilled off, crystals precipitate and are collected by filtration. Yield 2.1g, melting point
185℃ (decomposition), Rf ¹ = 0.20, [α] ²¹ _D +1.8゜(c
= 0.5, DMF), elemental analysis Calculated value as C ₂₁ H ₃₁ O ₈ N ₃ S・H ₂ O
C 50.08, H 6.20, N 8.35, S 6.36 Actual value
C 49.77, H 6.32, N 8.25, S 6.10. () BOC−Tyr−(D)−Met(O)−Gly−Phe−
Production of NH-NH-CO-O-CH ₂ -CH ₃ Z-Phe-NH-NH-CO-O-CH ₂ -
Dissolve 460 mg of CH ₃ in 50 ml of MeOH and perform catalytic reduction using palladium black as a catalyst. Filter out the catalyst
MeOH is distilled off and the residue is dissolved in 10 ml of DMF.
On the other hand, BOC−Tyr−(D)−Met(O)−Gly−OH485
Dissolve mg and 215 mg of HONB in 5 ml of DMF, cool to 0°C, add 230 mg of DCC, and stir at 0°C for 6 hours. Insoluble materials are filtered off, and the filtrate is combined with the amine component and stirred overnight at room temperature. DMF was distilled off and the residue was extracted with 50 ml of n-BuOH and washed with water. n-BuOH was distilled off and the residue was filtered with diethyl ether. Yield 630 mg, melting point 145-147°C, Rf ¹ = 0.28, [α] ²¹ _D -3.6° (c
= 0.5, DMF), elemental analysis Calculated value as C ₃₃ H ₄₆ O ₁₀ N ₆ S
C 55.13, H 6.45, N 11.69, S 4.46 Actual value
C 54.85, H 6.29, N 11.45, S 4.35. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO-O- _CH2 - _CH3 production BOC-Tyr-(D)-Met(O)-Gly-Phe-NH
The desired compound is obtained in the same manner as in Example 1 () using 350 mg of -NH-CO-O-CH ₂ -CH ₃ . Yield 205 mg, Rf ² = 0.39, [α] ²¹ _D +
15.0゜(c=0.38, MeOH), Amino acid analysis (acid hydrolyzate): Gly1.00, Met0.22, Tyr0.88,
Phe1.00. Example 9 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-CH ₂ -S-CH ₃ () BOC-Phe-NH-NH-CO-CH ₂ -S-
Production of CH ₃ Dissolve 1.4 ml of ethyl thioglycolate in 20 ml of acetonitrile and add 1 ml of methyl iodide.
Cool and add 2.1ml of TEA while stirring.
Stir at room temperature for 3 hours. Acetonitrile was distilled off and the residue was extracted with diethyl ether, washed with water and dried over anhydrous sodium sulfate. Diethyl ether was distilled off, the residue was dissolved in MeOH, 0.7 ml of hydrazine hydrate was added, and the mixture was allowed to stand at room temperature for 2 days. MeOH is distilled off and the residue is dissolved in 20 ml of DMF. Add 3.2 g of BOC-Phe-ONB to this and stir overnight at room temperature. DMF is distilled off and the residue is extracted with 100 ml of AcOEt, washed with water and dried over anhydrous sodium sulfate. AcOEt is distilled off and the precipitated crystals are collected by filtration. AcOEt - Recrystallize with petroleum benzine. Yield 2.3g, melting point 151-152℃,
Rf ¹ = 0.66, [α] ²¹ _D -18.6° (c = 0.5DMF), elemental analysis Calculated value as C ₁₇ H ₂₅ O ₄ N ₃ S
C 55.55, H 6.86, N 11.44, S 8.73 Actual value
C 55.59, H 6.89, N 11.23, S 8.33. () BOC−Tyr−(D)−Met(O)−Gly−Phe−
Production of NH-NH-CO-CH ₂ -S-CH ₃ BOC-Phe-NH-NH-CO-CH ₂ -S-
Dissolve 420 mg of CH ₃ in 4 ml of TFA and leave at room temperature for 20 minutes. TFA is distilled off and the residue is filtered with diethyl ether. Add this powder to DMF10
ml and add 0.16ml of TEA. On the other hand, BOC−Tyr−D−Met(O)−Gly−
Dissolve 485mg of OH and 215mg of HONB in 5ml of DMF.
Cool to ℃, add 230 mg of DCC, and stir at 0℃ for 6 hours. Insoluble matter is filtered off, and the filtrate is combined with the amine component prepared earlier and stirred overnight at room temperature. DMF is distilled off and solidified with diethyl ether. Yield 630mg, melting point 154-156℃, Rf ¹ =
0.25, [α] ²¹ _D -2.0゜ (c=0.46, DMF) Elemental analysis Calculated value as C ₃₃ H ₄₆ O ₉ N ₆ S ₂
C 53.93, H 6.31, N 11.43, S 8.72 Actual value
C 53.64, H 6.42, N 11.25, S 8.45. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO-CH ₂ -S-CH ₃ production BOC-Tyr-(D)-Met(O)-Gly-Phe-NH
Using 300 mg of -NH-CO-CH ₂ -S-CH _{3 ,} 220 mg of the desired product was obtained in the same manner as in Example 1 (). Rf ² = 0.42, [α] ²¹ _D +15.4° (c = 0.33,
MeOH), amino acid analysis (acid hydrolyzate):
Gly1.00, Met0.38, Tyr0.58, Phe1.02. Example 10 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-CH ₂ -SO-CH ₃ H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Dissolve 100 mg of NH-CO-CH ₂ -S-CH ₃ in 3 ml of N-acetic acid and add 30% hydrogen peroxide while stirring.
Add 0.1ml and stir at room temperature for 5 hours. The powder obtained by freeze-drying the reaction solution is dissolved in a small amount of 0.1N aqueous acetic acid, applied to a Sephadex LH-20 column (2.5 x 120 cm), and eluted with aqueous 0.1N acetic acid. The eluted fractions from 290 ml to 305 ml are collected and lyophilized. Yield 70 mg, Rf ² = 0.32, [α] ²¹ _D
+13.5° (c=0.46, MeOH), Amino acid analysis (acid hydrolyzate): Gly1.00, Met0.29, Tyr0.65,
Phe1.02. Example 11 H-Tyr-(D)-Met(O)-Gly-MePhe-NH
Production of -NH-CO-CH ₂ -CH ₃ () Production of BOC-(D)-Met-Gly-OH Dissolve 4.7 g of BOC-(D)-Met-Gly-OEt in 20 ml of ethyl alcohol and cool. -Add 16ml of caustic soda water and stir at room temperature for 1 hour. Ethyl alcohol is distilled off, the residue is neutralized by adding citric acid water, extracted with 100 ml of acetic acid ethyl ester, and washed with water. Dry with anhydrous sodium sulfate. AcOEt is distilled off and crystallized with petroleum benzene. AcOEt - Recrystallize with petroleum benzine.
Yield 3.8g, melting point 124-125℃, ^Rf1 = 0.41,
[α] ²¹ _D +12.3° (c=0.59, DMF), elemental analysis Calculated value as C ₁₂ H ₂₂ O ₅ N ₂ S
C 47.04, H 7.23, N 9.14, S 10.46 Actual values
C 46.79, H 7.45, N 9.20, S 10.21. () BOC−(D)−Met(O)−Gly−MePhe−NH
Production of -NH-CO-CH ₂ -CH ₃ Dissolve 4.7 g of Z-Me-Phe-OH in 50 ml of MeOH, add 2 ml of 6N-hydrochloric acid-dioxane, and leave at room temperature overnight. MeOH was distilled off and washed with water.
Extract with 100ml of AcOEt and wash with water. Dry with anhydrous sodium sulfate. AcOEt was distilled off, the residue was dissolved in 50 ml of MeOH, and 1.5 ml of hydrazine hydrate was added.
ml and leave at room temperature for 3 days. Distill MeOH off, extract with 100 ml of AcOEt, and wash with water. Dry with anhydrous sodium sulfate. Distill AcOEt and dissolve the residue in 20ml of THF with 1.4% propionic anhydride.
ml, cool, add 1.4 ml of TEA, and stir at room temperature for 5 hours. Distill the TEF and remove the residue
Extract with 100ml of AcOEt and wash with 5% dehydrated water. After drying with anhydrous sodium sulfate and distilling off AcOEt, an oily Z-Me-Phe-NH-NH-CO-
3.8 g of _CH2 - _CH3 are obtained. 2.3g of this
Dissolve in 50ml of MeOH and perform catalytic reduction using palladium black as a catalyst. The catalyst is filtered off, MeOH is distilled off and the residue is dissolved in 10 ml of DMF. On the other hand, 1.85 g of BOC-(D)-Met-Gly-OH
Dissolve 1.2g of HONB in 20ml of acetonitrile.
Cool to ℃, add 1.4 g of DCC, and stir at 0℃ for 4 hours. Insoluble materials are filtered off, and the filtrate is combined with the amine component and stirred overnight at room temperature. The solvent was distilled off and the residue was washed with 100 ml of AcOEt.
The AcOEt solution was concentrated to 20 ml, and 5 ml of acetic acid and 0.5 ml of 30% hydrogen peroxide were added to it, and the solution was heated to 5 ml at room temperature.
Stir the time. Add 50 ml of n-BuOH to the reaction solution for extraction, and wash with water. n-BuOH was distilled off and the residue was filtered with diethyl ether.
Yield 1.9g, melting point 116-117℃, Rf ¹ = 0.20,
[α] ²¹ _D -25.7° (c=0.38, DMF), elemental analysis Calculated value as C ₂₅ H ₃₉ O ₇ N ₅ S
C 54.22, H 7.09, N 12.65, S 5.79 Actual value
C 54.05, H 6.98, N 12.41, S 5.48. () BOC−Tyr−(D)−Met(O)−Gly−MePhe
-NH-NH-CO-CH2 _{- CH3} production BOC-(D)-Met(O)-Gly-MePhe-NH-
Dissolve 420 mg of NH-CO-CH ₂ -CH ₃ in 4 ml of TFA and leave at room temperature for 20 minutes. TFA is distilled off and the residue is filtered through diethyl ether and dried. Dissolve this powder in 10ml of DMF and cool.
Add 0.14ml of TEA. To this, BOC－Tyr－
Add 330 mg of ONB and stir overnight at room temperature. DMF
is distilled off, extracted with 50 ml of n-BuOH, and washed with water.
n-BuOH was distilled off and the residue was filtered with diethyl ether. Reprecipitate from methanol to ether. Yield 310mg, melting point 138-139℃,
Rf ¹ = 0.20, [α] ²¹ _D -21.3° (c = 0.55,
DMF), elemental analysis Calculated value as C ₃₄ H ₄₈ O ₉ N ₆ S
C 56.96, H 6.74, N 11.72, S 4.47 Actual value
C 56.58, H 6.48, N 11.58, S 4.23. () H-Tyr-(D)-Met(O)-Gly-MePhe-
Production of NH-NH-CO-CH ₂ -CH ₃ BOC-Tyr-(D)-Met(O)-Gly-MePhe-
Dissolve 150 mg of NH-NH-CO-CH ₂ -CH ₃ in 2 ml of TFA and leave at room temperature for 30 minutes. TFA is distilled off and the residue is filtered with diethyl ether. Dissolve this powder in a small amount of water and use an Amberlite IRA-410 (acetic acid type) column (2 x 4 cm).
The flow through and washing liquid are combined and lyophilized.
The obtained powder was purified in the same manner as in Example 1 () to obtain 60 mg of the desired product. Rf ² =0.33, [α] ²¹ _D +4.1
° (c=0.21, MeOH), amino acid analysis (acid hydrolyzate): Gly1.00, Met0.24, Tyr0.91. Example 12 H-MeTyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO- _CH2 - _CH3 production () Z-MeTyr( ^But )-(D)-Met(O)-Gly-
Production of Phe-NH-NH-CO-CH ₂ -CH ₃ BOC-(D)-Met(O)-Gly-Phe-NH-NH
−CO−CH ₂ −CH ₃ 470mg was dissolved in TFA4ml,
Leave at room temperature for 10 minutes. TFA is distilled off and the residue is filtered off with diethyl ether. Dissolve this powder in 10ml of DMF, cool, and 0.15ml of TEA.
and Z−MeTyr(Bu ^t )−ONB〔Z−MeTyr
(But) prepared from 380 mg of OH and 215 mg of HONB using the DCC method] and stirred overnight at room temperature. DMF is distilled off, and the residue is extracted with 100 ml of AcOEt, washed with water, and dried over anhydrous sodium sulfate. AcOEt is distilled off and the residue is filtered off with diethyl ether. Yield 450 mg, Rf ¹ =0.46, [α] ²¹ _D -30.0
゜(c=0.5, DMF), () H-MeTyr-(D)-Met(O)-Gly-Phe-
Production of NH-NH-CO-CH2 _{- CH3} Z-MeTyr-( ^But )-(D)-Met(O)-Gly-
Using 350 mg of Phe-NH-NH-CO-CH ₂ -CH ₃ and using the same method as in Example 2 (), 120 mg of the target product was obtained.
get. Rf ² = 0.34, [α] ²¹ _D +19.5° (c =
0.20, MeOH), amino acid analysis (acid hydrolyzate):
Gly1.00, Met0.32, Phe0.98. Example 13 H-MeTyr-(D)-Met(O)-Gly-MePhe-
Production of NH-NH-CO-CH ₂ -CH ₃ () Z-MeTyr( ^But )-(D)-Met(O)-Gly-
Production of MePhe−NH−NH−CO−CH ₂ −CH ₃ BOC−(D)−Met(O)−Gly−MePhe−NH−
Dissolve 550 mg of NH-CO-CH ₂ -CH ₃ in 4 ml of TFA and leave at room temperature for 10 minutes. TFA is distilled off and the residue is filtered through diethyl ether and dried. Dissolve this powder in 10ml of DMF and cool.
Add 0.16ml of TFA. Z-MeTyr to this
( ^But )-ONB [Z-MeTyr( ^But )-OH385mg
Add 210 mg of HONB (adjusted using the DCC method) and stir at room temperature overnight. Distill the DMF and remove the residue.
Extract with 100ml of AcOEt and wash with water. After drying over anhydrous sodium sulfate, AcOEt is distilled off and the residue is filtered through diethyl ether. yield
450mg, melting point 121-123℃, Rf ¹ = 0.48, elemental analysis Calculated value as C ₄₂ H ₅₆ O ₉ N ₆ S・H ₂ O
C 60.12, H 6.97, N 10.01, S 3.82 Actual value
C 59.88, H 6.86, N 9.75, S 3.91 () H-MeTyr-(D)-Met(O)-Gly-MePhe
Production of -NH-NH-CO-CH ₂ -CH ₃ Z-MeTyr( ^But )-(D)-Met(O)-Gly-
Using 250 mg of MePhe-NH-NH-CO- _CH2 - _CH3 , the target compound 105 was obtained in the same manner as in () of Example 2.
Get mg. Rf ² = 0.36, amino acid analysis (hydrochloric acid hydrolyzate): Gly 1.00, Met 0.31. Example 14 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO- _CH2 - _CH2- Cl. () Z−Tyr−(D)−Met(O)−Gly−Phe−NH
-NH-CO- _CH2 - _CH2 -Cl production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
The target product 410 was prepared in the same manner as in Example 2 () using 510 mg of NH ₂ and 100 mg of β-chloropropionic acid.
Get mg. Melting point 172-175℃, Rf ¹ = 0.26, [α]
²¹ _D -8.9° (c=0.50, DMF), Elemental analysis C ₃₆ H ₄₃ O ₉ N ₆ SCl・H ₂ O Calculated values C 54.78, H 5.74, N 10.65,
S 4.06, Cl4.49 Actual value C 54.45, H 5.62, N 10.54,
S 4.12, Cl 4.03. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO- _CH2 - _CH2 -Cl production Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
120 mg of the target compound is obtained in the same manner as in Example 2 () using 300 mg of NH-CO-CH ₂ -CH ₂ -Cl. Rf ² = 0.37, [α] ²¹ _D +25.4° (c = 0.40,
MeOH), amino acid analysis (acid hydrolyzate):
Gly1.00, Met0.40, Tyr0.92, Phe0.98. Example 15 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-CH ₂ -CH ₂ -CO-CH ₃ () Z-Tyr-(D)-Met(O)-Gly-Phe-NH
Production of -NH-CO-CH ₂ -CH ₂ -CO-CH ₃ Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
Using 510 mg of NH ₂ and 110 mg of levulinic acid, 380 mg of the target compound was obtained in the same manner as in Example 2 (). Melting point 161-162℃, Rf ¹ = 0.25, [α] ²¹ _D −
7.6゜ (c=0.35, DMF), elemental analysis Calculated value as C ₃₈ H ₄₆ O ₁₀ N ₆ S・H ₂ O
C 57.27, H 6.07, N 10.54, S 4.02 Actual value
C 57.05, H 6.12, N 10.23, S 3.85. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
Production of -NH-CO-CH ₂ -CH ₂ -CO-CH ₃ Z-Tyr-(D)-Met(O)-Gly-Phe-NH-
Using 350 mg of NH-CO- _CH2 - _CH2 -CO- _CH3, 130 mg of the target compound is obtained in the same manner as in Example 2 (). Rf ² = 0.35, [α] _D +26.0° (c = 0.25, MeOH), amino acid analysis (acid hydrolyzate): Gly1.00, Met0.33, Tyr0.98,
Phe1.02. Example 16 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of NH-CO-CH ₂ -CH ₂ -OH () BOC-Tyr-(D)-Met(O)-Gly-Phe-
Production of NH-NH-CO-CH ₂ -CH ₂ -OH Z-Phe-NH-NH-CO-CH ₂ -CH ₂ -
OH460mg and BOC-Tyr-(D)-Met(O)-Gly-
Using 485 mg of OH, 620 mg of the target compound was obtained in the same manner as in Example 8 (). Melting point 148−
150°C, Rf ¹ = 0.20, [α] ²¹ _D -2.6° (c = 0.40,
DMF), elemental analysis Calculated value as C ₃₃ H ₄₆ O ₁₀ N ₆ S
C 55.13, H 6.45, N 11.69, S 4.46 Actual value
C 54.86, H 6.25, N 11.42, S 4.19. ()H-Tyr-(D)-Met(O)-Gly-Phe-NH
-NH-CO- _CH2 - _CH2 -OH production BOC-Tyr-(D)-Met(O)-Gly-Phe-NH
Example 1 -NH-CO- _CH2 - _CH2 -OH350mg
150mg of the target substance was processed in the same manner as ().
get. Rf ² = 0.30, [α] ²¹ _D +19.4° (c =
0.35, MeOH), amino acid analysis Gly1.00,
Met0.32, Tyr0.89, Phe1.00. Example 17 H-Tyr-(D)-Met(O)-Gly-Phe-NH-
Production of _NH2 () H-Tyr-(D)-Met(O)-Gly-Phe-NH
-Production of NH ₂ Z obtained by the same method as in Example 2 ()
-Tyr-(D)-Met(O)-Gly-Phe-NH-
Using 300 mg of NH ₂ , 140 mg of the target compound was obtained in the same manner as in Example 2 (). ^Rf2 =
0.20, [α] ²¹ _D +33.0° (c=0.35, MeOH), amino acid analysis (acid hydrolyzate): Gly1.00, Met0.29,
Tyr0.95, Phe1.00. Example 18 H-Tyr-(D)-Met(O)-Gly-MePhe-
Production of NHNH-CO- _CH3 () Production of BOC-(D)-Met(O)-OH Dissolve BOC-(D)-Met-OH (37g) in acetonitrile (200ml) and 30% hydrogen peroxide solution ( 19
ml) and stir at room temperature for 8 hours. Add palladium black and stir for another hour. Filter off palladium black, distill off acetonitrile,
Extract with 300 ml of n-butanol. Wash with brine and distill off n-butanol. Crystallize from ether and recrystallize from AcOEt-ether;
32g. Melting point 125-126℃, [α] ^23D _+7.2゜(c=
0.5, DMF), Rf ¹ = 0.27 Elemental analysis Calculated value as C ₁₀ H ₁₉ O ₅ N ₁ S:
C 45.26, H 7.21, N 5.27, S 12.08 Actual values:
C 45.31, H 7.27, N 5.01, S 11.83 () BOC-(D)-Met(O)-Gly-MePhe-
Production of NHNH-CO- _CH3 Z-MePhe-OH (3.1 g) and HONB (1.9
g) in THF (20 ml), cooled and DCC
(2.3g) and stir at 0℃ for 4 hours. Filter out insoluble materials and add NH ₂ NHCOCH ₃ (0.9 g) to the filtrate.
Add and stir at room temperature for 3 hours. THF was distilled off, extracted with AcOEt (100ml), washed with water and anhydrous.
Dry _{with Na2SO4} . AcOEt is distilled off, the residue is dissolved in MeOH (100 ml), and catalytic reduction is performed using palladium black as a catalyst. Filter the catalyst,
Distill MeOH and filter the precipitated crystals;
2.0g. Of this, 1.3g and BOC-Gly-ONB (2.0
Dissolve g) in acetonitrile (30 ml) and stir at room temperature overnight. Distill the solvent and remove the residue
Extract with AcOEt (100ml) and wash with water. anhydrous
After drying with Na ₂ SO ₄ , AcOEt was distilled off to obtain 2.3 g of oil. Dissolve this in TFA (20 ml) and leave at room temperature for 20 minutes. After distilling off TFA, add ether to the residue and wash thoroughly with decant. to this
Add DMF (20ml), cool and add TEA (0.8ml)
and BOC−(D)−Met(O)−ONB(BOC−(D)−
(Prepared by DCC method from 1.3 g of Met(O)-OH and 1.0 g of HONB) and stirred overnight at room temperature. DMF
is distilled off, extracted with n-butanol (100 ml), and washed with water. The n-butanol was distilled off and the residue was solidified with ether. Reprecipitate with methanol-ether; 1.5 g. Melting point 119-121℃, [α] ^24D _-29.0゜(c=0.38,
DMF), Rf ¹ = 0.19 Elemental analysis C ₂₄ H ₃₇ O ₇ N ₅ S・H ₂ O Calculated value:
C 51.69, H 7.05, N 12.56, S 5.05 Actual values:
C 51.64, H 7.18, N 12.11, S 5.76 () BOC−Tyr−(D)−Met(O)−Gly−MePhe
-NHNH-CO- _CH3 production BOC-(D)-Met(O)-Gly-MePhe-NHNH
−CO−CH ₃ (0.54 g) and BOC−Tyr−ONB
(0.46 g) in the same manner as in () of Example 11 using (0.46 g). Melting point 141-142℃, [α] ^22D _-25.4゜(c=0.48,
DMF), Rf ¹ = 0.22 Elemental analysis C ₃₃ H ₄₆ O ₉ N ₆ S・2H ₂ O Calculated value:
C 53.64, H 6.81, N 11.37, S 4.34 Actual values:
C 53.15, H 6.65, N 10.77, S 4.23 () H-Tyr-(D)-Met(O)-Gly-MePhe-
Production of NHNH-CO- _CH3 BOC-Tyr-(D)-Met(O)-Gly-MePhe-
Example using NHNH-CO-CH ₃ (0.35g)
Produced in the same manner as in 11 (); 0.19g,
[α] ²² _D 0° (c = 0.25, MeOH), Rf ² = 0.16,
Amino acid analysis values: Gly1.00, Met1.00, Tyr1.20. Example 19 H-MeTyr-(D)-Met(O)-Gly-Phe-
Production of NHNH-CO-CH ₃ () Production of Z-Phe-NHNH-CO-CH ₃ Add Z-Phe-NHNH ₂ (2.0g) to THF (50ml)
Dissolve, cool and add acetic anhydride (1.4ml) and TEA.
(0.9ml) and stir at room temperature for 2 hours. Filter the precipitated crystals and wash thoroughly with ether. Recrystallized from ethyl acetate; 1.9 g. Melting point 205-206℃, [α] ^23D _-16.4゜(c=0.50,
DMF), Rf ¹ = 0.60. Elemental analysis As C ₂₀ H ₂₃ O ₅ N ₃ Calculated values: C 62.32, H 6.02, N 10.90 Actual values: C 62.38, H 6.09, N 10.49 () Z−Gly−Phe−NHNH Production of -CO-CH ₃ Z-Phe-NHNH-CO-CH ₃ (2.1g) and Z
Produced in the same manner as in Example 1 () using -Gly-ONB (2.2 g). Melting point 154-155℃, [α] ^23D _-0.9゜(c=0.50,
DMF), Rf ¹ = 0.52 Elemental analysis as C ₂₁ H ₂₄ O ₅ N ₄ Calculated values: C 61.15, H 5.87, N 13.59 Actual values: C 60.98, H 5.68, N 13.27 () BOC−(D)−Met() O)-Gly-Phe-NHNH
Production of -CO-CH ₃ BOC-(D)-Met-OH (0.60g) and Z-Gly-
Produced in the same manner as in Example 1 () using Phe-NHNH-CO-CH ₃ (1.0 g), 1.0
g. Melting point 122-123℃, [α] ²¹ _D 0゜(c=0.5,
DMF), Rf ¹ = 0.38. Elemental analysis Calculated value as C ₂₃ H ₃₅ O ₇ N ₅ S:
C 52.56, H 6.72, N 13.32, S 6.09 Actual values:
C 52.35, H 6.62, N 13.12, S 5.93 () Z-MeTyr( ^But )-(D)-Met(O)-Gly-
Production of Phe-NHNH-CO- _CH3 BOC-(D)-Met(O)-Gly-Phe-NHNH-
CO- _CH3 (0.55g) and Z-MeTyr( ^But )-OH
Produced in the same manner as in Example 12 () using (0.39 g); 0.61 g, melting point 140-142°C, [α] ²¹ _D -29.0° (c = 0.5,
DMF), Rf ¹ = 0.43. Elemental analysis Calculated value as C ₄₀ H ₅₂ O ₉ N ₆ S:
C 60.59, H 6.61, N 10.59, S 4.04 Actual values:
C 60.28, H 6.55, N 10.28, S 3.88 () H-MeTyr-(D)-Met(O)-Gly-Phe-
Production of NHNH-CO- _CH3 Z-MeTyr( ^But )-(D)-Met(O)-Gly-
Using Phe-NHNH-CO-CH ₃ (300 mg), 110 mg of the desired product was obtained in the same manner as in Example 2 (). [α] ²¹ _D +18.5° (c=0.25, MeOH), Rf ²
=0.29. Amino acid analysis value, Gly1.00, Met0.35, Phe0.99 Example 20 H-MeTyr-(D)-Met(O)-Gly-MePhe-
Production of NHNH-CO- _CH3 ()Z-MeTyr( ^But )-(D)-Met(O)-Gly-
Production of MePhe-NHNH-CO- _CH3 BOC-(D)-Met(O)-Gly-MePhe-NHNH
−CO−CH ₃ (0.50 g) and Z−MeTyr( ^But )−
0.40 g, prepared as in Example 13 () using OH (0.39 g). Melting point 120-122℃,
Rf ¹ = 0.46. Elemental analysis Calculated value as C ₄₁ H ₅₄ O ₉ N ₉ S:
C 61.02, H 6.74, N 10.42, S 3.97 Actual values:
C 60.79, H 6.82, N 9.92, S 3.81 () H-MeTyr-(D)-Met(O)-Gly-MePhe
Production of -NHNH-CO- _CH3 Z-MeTyr( ^But )-(D)-Met(O)-Gly-
Produced in the same manner as in Example 2 () using MePhe-NHNH-CO-CH ₃ (0.02g); 0.90
g. ^Rf2 =0.34. Amino acid analysis values: Gly1.00, Met0.38.

Claims (1)

[Claims] 1. General formula [In the formula, R ₁ and R ₂ each represent hydrogen or methyl, and R ₃ represents hydrogen, or lower aliphatic acyl which may be substituted with a hydroxyl group, lower alkoxy, lower alkylthio, lower alkylthiooxide, halogen or oxo group. , respectively. ]
Tetrapeptide derivatives represented by: and pharmacologically acceptable acid addition salts thereof. 2 General formula [In the formula, R ₁ and R ₂ each represent hydrogen or methyl, and R ₃ ' represents hydrogen, or lower aliphatic acyl optionally substituted with a hydroxyl group, lower alkoxy, lower alkylthio, lower alkylthiooxide, halogen, or oxo group. and may have a protecting group, Y ₁ represents a protecting group, and Y ₂ and Y ₃ represent hydrogen or a protecting group, respectively. A general formula characterized by subjecting a compound represented by ] to a protecting group elimination reaction [In the formula, R ₁ and R ₂ have the same meanings as above. _R3
represents hydrogen, or lower aliphatic acyl optionally substituted with a hydroxyl group, lower alkoxy, lower alkylthio, lower alkylthiooxide, halogen or oxo group. ] A method for producing a tetrapeptide derivative represented by.