JPH02233694A - Peptide compound and use thereof - Google Patents

Abstract

NEW MATERIAL:A compound shown by formula I [X is D-Ala, D-Met(O) or D-Arg; Y is Phe, Phe(NO2) or MePhe; Z is group shown by formula II, Leu-NH- CH2 or NH-CH2]. EXAMPLE:A compound shown by formula III. USE:Having opioid activity, manifesting morphine-like analgesic action irrespective of dosage form and useful as an analgesic. PREPARATION:A compound shown by formula IV is dissolved in methanol, mixed with (n-Bu)3P while stirring, further with a solution of Npys-Cl in dioxane at 0 deg.C and reacted at room temperature and powder is obtained from the filtrate and recrystallized to give the aimed compound.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention relates to novel peptide compounds, acid addition salts and hydrates thereof, and preparations containing at least one of these as an active ingredient, especially The present invention relates to an analgesic composition.

(Prior Art) Exogenous opiates such as morphine have a strong analgesic effect, but have problems in that they are accompanied by side effects such as habit-forming and addictive properties.

In the early 1370s, it was demonstrated that receptors for opioids existed in the brains of odontocetes, and many research groups attempted to search for endogenous morphine-like substances. As a result, in 1975!
Iughes et al.
phalln) was discovered as an endogenous morphine-like peptide [Hughes et al. ”Nature
"258, 577 (1975)]. Furthermore, Enkef 1
A lot of research has been conducted to search for phosphorus analogs and to elucidate the relationship between structure and activity, and as a result, β-endorphin was discovered. Since this β-dolphin is a polypeptide composed of 31 amino acids, it is extremely difficult to manufacture, and therefore it is difficult to mass-produce it for use as a medicine. On the other hand, regarding the research status of the synthetic chemical cephalin, there is a review article by Shimohigashi et al., one of the inventors of the present invention [
Yasuyuki Shimohigashi, Noriwaki Waki, Nobuo Izumiya, “Protein Nucleic Acid Enzyme” 2
8. 1321 (1983)], but its development was: a) a low-molecular compound that is easy to manufacture; b)
) High receptor affinity% C) High resistance to hydrolytic enzymes such as aminopeptidase and carboxypeptidase% a) Easy absorption and diffusion e) Capable of passing through the blood-brain barrier r ) It is possible to administer orally, and g) it has few side effects.The aim is to have enkephalin as a parent substance and consider amino acid substitution, modification of amino acid residues, shortening, cyclization, etc. It was something.

However, the reality is that no compound that is fully satisfactory has been found. Therefore, as a result of further various studies, the present inventors have already discovered several suitable peptide compounds (for example, JP-A No. 8
3-58599 and 63-128892).

(Problem to be Solved by the Invention or Object of the Invention) The main object of the present invention is to develop a novel peptide compound that is relatively easy to produce, has obioid activity, and can therefore be used as an active ingredient of analgesics. It is about providing.

An additional object of the present invention is to provide a novel peptide compound whose analgesic effect is independent of dosage form.

(Means and actions for solving the problem and achieving the object) According to the present invention, the above object is achieved by the formula NOI [wherein, X is D-▲la, D-get(0) or D-▲ rg, Y is Pbe, Phe (NOa) or )
means IePhe, 2 is Nu-CB, Leu-I
II-11: H* or NH-CTo with CH*-Cll(
CL) *means] This is achieved by the peptide compound represented by CL)* and its pharmacologically acceptable acid addition salts and hydrates.

The peptide compound according to the present invention can be produced according to the method described in the production example below, or can be produced using a general-purpose peptide synthesis method, for example, "Fundamentals and Experiments of Peptide Synthesis" by Izumiya et al.
(published by Maruzen Co., Ltd., 1985).

The acid addition salt of the compound according to the present invention is a salt that is pharmacologically acceptable in relation to its use, and examples of the salt-forming acid include inorganic acids such as hydrochloric acid, sulfuric acid, sulfonic acid, and phosphoric acid. Organic acids such as citric acid, benzoic acid, acetic acid, propionic acid, lactic acid, maleic acid, conolic acid, and tartaric acid can be mentioned.

When formulating at least one of the peptide compounds, acid addition salts, and hydrates according to the present invention as a therapeutic ingredient, there are no restrictions on the dosage form. Tablets for oral administration;
They can be formulated as capsules, powders, granules, suspensions or solutions, and in the case of parenteral administration, as sterile solutions or suspensions.

Note that, as a matter of course, excipients, lubricants, preservatives, and stabilizers can be used during formulation.

The dosage of the compounds according to the invention is determined taking into account the type and extent of the disease, the age and weight of the patient and various other factors.

(Examples, etc.) Next, the present invention will be described in further detail with reference to production examples, pharmacological tests, etc.

The abbreviations used in this specification, including those in this section, are in accordance with the abbreviations commonly used in the field,
The relationship between each abbreviation and its meaning is as follows; in principle, amino acids are L-units, and rDJ is added when specifically referring to D-units.

Ala: Alanine Phe: Phenylalanine Phe (NOa): 4-nitrophenylalanine IePhe: N-methylphenylalanine κe
t(0)' Methionine sulfoxide Leu
: Leucine ▲rg : Arginine Tyr : Tyrosine cry : Glycine Boc : t-butyloxycarbonyl Gly
ol: Glycineol DIIIF: Dimethylformamide TFA
:}Lifluoroacetic acid Npys: 3-nitro-2-pyridinesulfenyl (Boa-Tyr-D-Ala-Gly-Phe-N
IICH2CII*S-)a (2fi2.OB,
0.213 mmol) with methanol (81) and water (2
1), (n-Bu)aP (0.055+al
, 0.213i+mol) was added with stirring. 2
After 2 hours, Npys-CI (97.5 g, 0
．． 213 mmol) of dioxane solution (B1) was added with stirring. The reaction was carried out at 0° C. for 30 minutes and then at room temperature for 3 hours. Insoluble matter was filtered off, and the solvent was removed from the filtrate under reduced pressure. The residue was solidified by adding water, filtered, and dried on P206 in a desiccator. The obtained powder is DMF
(1ml) and added to a Sephadex L 20 column (φ3.8 x 40cm) and eluted with the same solvent. Fraxilon containing the target product was collected and the solvent was removed under reduced pressure. Crude crystals were obtained from the residue using ether.

This crude crystal was recrystallized from the same solvent (211.4 g
, yield; B4%).

Contains Hoe-Tyr-D-Ala-Gly-Phe-NII
CH*CR*S-Npy+ (85.2mg, 0.13
8 mmol) was dissolved in TFA and kept at 0° C. for 30 minutes, then the solvent was removed under reduced pressure. The residue was diluted with 30% AcOH.
(lsl), applied to a Sephadex G-15 column (φ2.O x 138 cm), and eluted with the same solvent. Fluxene containing the target product was collected and repeatedly freeze-dried with water to obtain the target product (82.1 g, yield: 94%).

4m and "ε" 11-Tyr-D-A la-G Iy-Phe-Le
u-NHC■acHas-Npys*▲cOB and H-
Tyr-D-▲la-Gly-Phe-Leu (CI1
*S-Npys) Synthesis of Ac011 Synthesis was performed in the same manner as in Production Example 2.

The physical constants of the materials according to the present invention synthesized according to the above production examples are shown in Tables 1 and 2 below.

赳 It was synthesized by the same procedure as in Production Example 1.

1” Rfs i CBCIs: Me011
: ▲cOH = 8 : l : 1
Carrier: Kieselgel Go (BPTLC), manufactured by Merck ■ Solvent system: Rf+; CHCIs: 1IeOH
: 9 : IRfa j CHCII :
leOH: AcOH: 50:
1G: 2 solvent system: Rf+; n-Bu011: AcOH: H snow 0: 4: 1: 5 (upper layer); n-BuO
H:▲c011: Viridine:H,0=4:ll
2; 0.1%▲coil: n-BuOH: Pyridine: 11: 5: 3 (upper layer) Carrier: Kieselgel 80 (HPTLC), manufactured by Merck & Co., Ltd. <<1> Sample a> Specimen sample book Substances according to the invention (T of the compounds in Preparation Examples 6 and 2)
F▲ salt) was used.

b) Control sample The following two types of materials were used.

1) DiGO H-Tyr-D-▲la-CIF-NMePhe-Gl
yol2) DAOLE H-Tyr-D-▲la-Gly-Phe-D-Leu
-011 (2) Type and method of test The following two types of tests were conducted.

Rh Rh ▲》Guinea pig isolated ileal contraction inhibition test (GPI test) Method such as κostert+tz [Kosterlitz
at al. ,”Br. J. Pharmacol
．． '''39, 398 (1970)]. That is, the carotid artery of a IIartleF male guinea pig was cut to death by exsanguination, the abdomen was opened, and a portion of the ileum (approximately 10 cm from the ileocecal region) was cut. A section spanning approximately 40 cm) was excised.The excised section was immediately treated with low Mg
” modified Krebs-bl carbon
ate buffer (118mM NaCl, 4.75mM
KCI, 2.54mM CaClt, 1.19aM
KIIIPO4, O. 12mM MgSO
4, 25mM NallCOs, I1sai
After thoroughly washing and removing the contents in 1glucose), the method of Paton et al. [Paton et al. ．． ”
J. Phys1o1. (London)”194.
A longitudinal muscle specimen with nerve plexus was prepared according to [13 (19H)] and cut to a length of approximately 2-3 cm.

The above muscle specimen was made into a ring bar, and the lower end was placed on a platinum wire electrode fixed at the bottom of the bath.
The above buffer solution (35°C) through which mixed gas of CO* was aerated7
Rectangular wave electrical stimulation (70V, 0.IHZI 1 m
Bec) was given. The contraction caused by this electrical stimulation was recorded through a transducer. This contraction is 50
The concentration (ICio) of each sample required for % inhibition was determined and used as the titer of the morphine-like activity of the sample.

B》Mouse ring vas deferens contraction inhibition test (MVD test)H
Ughes et al.'s method [1] Ughes et a+,,
"Br. J.Pharsacol." 53, 3
71 (l9f5)].

Specifically, ddY mice were decapitated and exsanguinated to death, and the ring vas deferens was immediately removed. The removed ring vas deferens was immediately treated with Mg” fr
ee modified Krebs-blcarbo
nate buffer (118d NaCl, 4.75
d KCI, 2,45i+M CaCI*.
1.19-Mκfl2PO4 r 25s+M Na
flGO3+ llsM gnorecose), the attached adipose tissue and blood vessels were removed as much as possible, and the contents were sufficiently washed with the above buffer solution to prepare a specimen.

The above specimen was kept at 35℃ and exposed to 95% 02 and 5% CO*.
Suspended in the buffer solution 7-1 to which a mixed gas of is aerated,
Square wave electrical stimulation (70V10.1Hz11asec)
was applied transmurally, and the contraction caused by this electrical stimulation was recorded through a transducer. The concentration (ICio) of each sample required to inhibit this contraction by 51% was determined to evaluate the morphine-like activity of the sample.

(3) Results and Discussion The test results for Tests A and B are shown in Table 3 below. From the results shown in Table 3, the substance of the present invention inhibited the contraction of the guinea pig ileum and the mouse vas deferens caused by electrical stimulation. Therefore, the substance of the present invention is considered to have mainly analgesic activity similar to that of morphine.

(1): H-Tyr-D-▲la-Gly-Phe
-Leu(CllaS)-Npys(TF▲Salt》 (2): H-Tyr-D-▲la-CIY-
Phe-NHCIIsCH*S-NpFs (TF▲ salt) Example gelatin l. Hard capsule No. 2 was filled with a mixture consisting of the following components to obtain capsules.

11-Tyr-D-Ala-Gly-Phe-Leu(
CII2S)Np7g (substance according to the invention)
50mg wheat starch
150 mg (Effects of the Invention) The peptide compounds according to the invention have opioid activity and therefore exhibit morphine-like analgesic action. The analgesic effect of the compounds of the present invention does not depend on the dosage form, and since they can be produced relatively easily according to general-purpose peptide synthesis methods, mass production is also possible.

Therefore, the present invention provides a compound that is extremely suitable for use as a therapeutic ingredient in pharmaceutical analgesics.

Claims (2)

[Claims] (1) Formula▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, X is D-Ala, D-Met(O) or D-A
rg, Y is Phe, Phe(NO_2) or M
means ePhe, and Z means ▲ ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ or NH-CH_2] Peptide compounds represented by, and pharmacologically acceptable acid addition salts and hydrates thereof. (2) Formula▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, X is D-Ala, D-Met(O) or D-A
rg, Y is Phe, Phe(NO_2) or M
ePhe, and Z means ▲Mathematical formula, chemical formula, table, etc.▼ or NH-CH_2] Among the peptide compounds shown in , and their pharmacologically acceptable acid addition salts and hydrates. An analgesic composition characterized by containing at least one substance as an active ingredient.