DD272227A1 - PROCESS FOR PREPARING HEART-EFFECTIVE PEPPITATE BASED PRAEPARES - Google Patents

Abstract

The invention relates to a process for the preparation of peptide-based herpes supplements suitable for the treatment of cardiovascular diseases. The substances produced according to the invention contain as active ingredients peptides of the casomorphine type and their derivatives. They strengthen the strength developed during the contraction of the heart muscle, normalize the heart rhythm and the coronary blood circulation and are characterized by good tolerability. The invention makes it possible to therapeutically influence states of cardiac insufficiency, cardiac ischemia and cardiac arrhythmia.

Description

Process for the preparation of cardiac peptide-based preparations

Field of application of the invention

The invention relates to a method for producing cardiopulmonary preparations containing as active component linear, cyclic or oyclisoh-branched oasomorphine-type peptides and derivatives. These compounds strengthen the strength of the heart, normalize the rhythm of the heart and contribute to improved blood flow to the uteri. The peptides are administered or infused intravenously, intraperitoneally or orally in a corresponding preparation using suitable adjuvants. By this invention, it is possible to have preparations available which effectively and with high compatibility improve the symptoms of heart failure and angina peotoris. The invention is suitable for use in medicine.

Chaittkteistik aer known technical solutions

To medically influence the heart failure and conditions of cardiac isobiaemia and cardiac arrhythmia r / enden a number. known. Medicines used (General and Special Pharmacology and Toxicology, edited by W. I'orth, D. Hensohler and W-, Ruimnel, 4 »Edition, Bibliographic« Institute Mannheim / Wif / n / ZUrioh., BI · Wissenschaftsve.ülag 1983, Page 192 260; Medi ζ ri so he

Pharmacology, edited by P. Markwardt, H. Matthias and W, Oelsner, VBB Georg Thieme Verlag Leipzig 1985, pages 441 to 533) · In the foreground of the cardiotonic effects are the effects of cardiac glycosides (digitoxin and g-strophanthin) and phosphodiesterase Henmers (Caffeine, Trapidil, Amrinone) Antianginous effects are achieved by reducing cardiac work and by improving blood circulation, with preference being given to nitro compounds, β-receptor bulblers and a number of other synthetic compounds. Antiarrhythmic effects are mainly achieved by drugs that affect ion currents through the heart membrane or react with adrenergic receptors. These substances are either alkaloids or glycosides of plant origin or synthetic compounds · Their isolation from natural sources, or their chemosynthesis is usually associated with a variety of reaction stages with precise analytical control and thus very expensive · The effects are essentially only against one directed pathophysiological change. The therapeutic range is sometimes low (eg, cardiac glycosides). Antiarrhythmic drugs predominantly have a negative inotropic effect. Disproportionate drugs with peptide structure have not been reported to be effective. However, a positive inotropic effect of Glukagou, Aagiotenain, Bradykinin, Urotensin II, boGtinmiter is known neuroaktis peptides and peptides from sea an-cones, which are not suitable for use on the heart due to their side effects (Parah, A. and Tuttie, R .: J. Pharmaool, Exper. Ther., 129 (1960) 49; Gibaon. · ·, Wallace, P. and Berne, H "A .: Gen · Comp. Endoorinol · 64 (1986), 435; Pla'con, E.3., Ref'jum, H. and Hotvedt, B .: J Cardiovaso · Phaxtnaool.3 (196) 459; Grimmelikhuijzen, CJP and Graff, D .: Proc Natl. Aoad., USA 83. (1986), 9817} Lloyd, Ρ · Ε .: Trends ffeuro Sei 2 (1986) , 428),

In addition, Dax-Uber known that β-Casomorphln-5 (Tyr-Pro-Phe-Iro-01: r) has eir.f / n bradycardic effect (Wei., ET, Lee, A. and Chang, Ϊ .Κ .: Life Scianoes 26, 1517 (1980)).

Object of the invention

It is an object of the invention to provide preparations based on peptides which, when administered, economize on cardiac activity by increasing the force of contraction, normalizing the sole frequency and improving blood circulation, are clearly chemically defined and are highly compatible with the active substance contained, in comparison with the previous ones Preparations can act more universally and can optionally be combined with known active ingredients. In addition to previously known possibilities, the heart activity can be promoted.

Explanation of the essence of the invention

- Task

The invention has for its object to develop methods for the production of cardioprotective preparations, which promote heart activity and cardiac output.

Features of the invention

According to the invention the object is achieved in that linear, cyclic or cyo-branched peptide analogues of Cosomor- * phin type, their salts or C- and / or N-terminally modified derivatives, preferably with primary or secondary aliphatic, aromatic or heterocyclic In the basic structure, Tyr-Pro-Phe-Pro-Gly Proline was also synthesized in one or both positions by hydroxyproline, dehydroproline, homoproline, pipecolic acid, alanine, phenylalanine, valine, leucine, isoleucine, lysine, ornithine , 2,4-diaminobutyric acid, arginine, cysteine, methionine, serine, threonine and phenylalanine are replaced by homophenylalanine, N-methylphenylalanine, tyrosine, N-methyltyrosine, the corresponding ring-substituted compounds, tryptophan, thyronine, thyroxime, valine, leucine, isoleucine and alanine could be. The amino acids can be used both in the L or D configuration or as a racemate in the peptide tetramers.

Glycine may or may not be present. If present, it may be replaced by any amino acid in the L or D configuration, or you may be exchanged as a Raoemat. In addition, the corresponding N-terminal tripeptides may be termed Y. / irkkomponente be used. Some preferred compounds are, for example, the oligopeptides Tyr-Pro-D-Phe-Pro-Gly, Tyr-D-Ala-Phe-D-Pro-Gly, Tyr-Pro-Phe-Phe-Gly-NH ₂ , Tyr-Pro. Val-Pro-Gly, Tyr-Pro-D-Val-Pro-Gly and Tyr-D-Met-Phe-Pro-NHg, The active components used have, inter alia, the following advantages:

ι · Non-toxic structures.

2. Possible activity poling and prolongation by incorporation of D-amino acids.

3. Increased resistance to enzymatic degradation.

4. Modification of the spectrum of action with simultaneous effect differentiation through targeted structural modifications.

5. Effects on Lipophilicity and Other Physicochemical Parameters in the Improvement of Transport and Penetration Properties by Targeted Structural Modifications ·

One of the preferred compounds, as stated above, the peptide of formula (I):

H-Tyr-Pro-D-Phe-Pro-Gly-OH (I)

The construction of the peptides can take place both by segment condensation and by stepwise extension of the peptide chain from the C-terminal end.

The procedure of the example of the peptide of the formula (I) is explained.

The strategic variant of the step-by-stcp condensation is characterized in that a GlycinderivaS (H-GIy-Z) stepwise with X-Pro-Y, XD-Phe-Y, X-Pro-Y and X-Tyr (T) -Y according to the coupling methods customary in peptide chemistry (compare E. Wünsch, Synthesis of Peptides in Houben-Weyl, Volume 15/11, Methods of Organic Chemistry, edited by E. Mueller,

Georg ThJ.eme Verlag Stuttgart 1974). In the first place, a fully-protected peptide of the formula (II) is obtained:

X-Tyr-Pro-D-Phe-Pro-Gly-Z (II)

Z is OH, methyl, ethyl, tert-butyl, benzyl, 4-nitrobenzyl and Phenaoylester,

X stands for tert-butyloxycarbonyl, p-methoxy-benzylosyoarbonyl, B.iphenylisopropyloxyoarbonyl, u _t oc '-dimethyl-3' 5-dlmethoxybenzyloxycarbonyl, o-nitrophenylsulfenyl, 9-fluorenylmethyloxycarbonyl,

Y is OH, pentafluorophenyl, pentachlorophenyl, trichloropheayl, 4-nitrophenyl, N-hydroxysuooinimide ester, N-hydroxy-5-norbornene-2,3-dicarboximide and hydrazide

T is H, tert-butyl, benzyl, benzyloxyoarbonyl, tert-butyloxyoarbonyl ·

After purification of the fully protected peptide of the formula (II) by Absorptionsohromatographle (in a silica gel column by chloroform / methanol elution or by gel filtration on Sephffdex LH-20 or LH-60 (eluent: methanol or dioxane) Sohutzgruppen with those in peptide chemistry usual Deblocklerungsverfahren (see E. Wünsch, see), in particular by means of HCl / dioxane, HCl / glacial acetic acid, HBr / Elsessig, trifluoroacetic acid, catalytic hydrogenation, transfer hydrogenolysis, zinc / glacial acetic acid, piperidine / dimethylformamide or alkaline hydrolysis and the resulting Peptid der Pormel CE) preferably isolated as hydrochloride · In the segmental linkage, which is characterized in that, for example, a dopeptide of the formula (III):

X-Tyr-Pro-Y (III)

with the tripeptide derivative of the formula (IV):

H-D-Phe-Pro-Gly-Z (IV)

preferably conjugated with the peptides in the peptide chemistry Ubliohen coupling methods, (see E, Wünsch, see above) via the mixed anhydride or the N, N'-Dioyolohexyloarbodiimid / 1-hydroxybenzotriazole method to peptides of the formula (II). X, T, Y, Z as defined above, after purification of the protected peptide according to formula (II), deprotection, preferably by aolysis or hydrogenolysis, and repeated column chromatography on silica gel with a chloroform / methanol gradient becomes the peptide of formula (I) received.

The obtained casomorphin-type peptide-based herpes-containing preparations are to be used in an appropriate preparation using suitable auxiliaries for a given medical indication at a dose range of 1 μg / kg to 1 mg / kg after intravenous, intraperitoneal or oral administration or infusion.

The invention will be explained in more detail with reference to exemplary embodiments, but not be limited thereby.

embodiments Abkttrzungsverzeiohnis:

Amino acid symbols according to IUFAC-IUB Commission on Bioohemioal Nomenclature _} J. Biol. Chem. 247 »977 (1972).

AoOH Boc CAIBE DC

BEPP

HOBti.Vak.

MeOHMin.

NEMOMeONBOTcpPERTSC

TEA THP Zers.

acetic acid

tert. Butyloxyoarbonyl

isobutylchloroformate Dunnohiohto-chromatogram _f -ohromato ·

graphic

ethyl acetate melting point

hours

N-hydroxybenzotriazole

in a vacuum

solvent

methanol minutes

N-ethylmorpholine Methylester

p-nitrobenzyl

trichlorophenyl petroleum ether room temperature Column chromatography, -omatochrome

graphic

triethylamine tetrahydrofuran decomposition By "usual work-up" is meant:

After completion of the coupling reaction, the respective crude product is taken up in ethyl acetate and the solution successively washed twice with water (NaCl saturated), three times with 5 parts KHSO.-solution, twice with water, three times with saturated sodium bicarbonate solution and three times with water.

7 2 2 27 ^!

sohen »The organic phase becomes Na ₂ SO. dried and the crude product by evaporating the solvent in vacuo isolated ·

The following solvent systems (in volume fractions) for thin-film chromatography on silica gel precast panels (SiIufol UV 254, CSSR) were used:

BAE benzene-acetone-acetic acid 50 + 20 + 1 BAW 2-butanol-formic acid-water 75 + 13.5 + 11.5 BEWE 1-Butanol-acetic acid-Wctsser-Essigiester

20 + 20 + 20 +

BPEW I-butanol-pyridine-acetic acid-water

30 + 20 + 6 +

C M chloroform-methanol 90 + 10 EPEY / ethyl acetate-pyridine-acetic acid-water

90 + 15 + 4.5 + 8.3

Example 1: Synthesis of H-Tyr-Pro-D-Phe-Pro-Gly-OH.HCl

Variant I: Segment condensation 1.1. Boo-Pro-Gly-ONB

3.23 g Boo-Pro-OH were dissolved in 50 ml of EA and treated at -15 ⁰ C successively with 2.1 ml of TEA and 1.95 ml CAIBE. After 10 min., 3.7 g of H-Gly-ONB.HCl and 2.1 ml of TEA were added to the reaction mixture and allowed to stir for 1.5 h at -15 ⁰ C and a further 12 h at RT. The batch was worked up as usual and the crude product recrystallized from diisopropyl ether.

Yield: 5.5 g (90 % of theory)

Mp: 92-93 ⁰ C ΒΊ ² I => -69.3 ° (c = 1, AoOH)

PC uniform in BAE, CM, EPEW

27221?

1.2. Boo-D-Phe-Pro-Gly-ONB

3.18 g Boo-D-Phe-OH, 1.68 ml TEA and 1.56 ml were dissolved at -15 ⁰ O in 40 ml of THF and NaOH GAIBE 10 min. With 4.13 g of H-Pro-Gly ONB · HCl (obtained duroh Entaoylierung of 1.1 using 1.1 N HCl / AcOH - Pp: 88-91 ⁰ C; ^ tJ ² J - -39.9 ° ο β 1, AcOH.) and 1.68 ml TEA as under 1.1. described reacted. It worked as Ublioh and recrystallized the crude product from EE / diisopropyl ether. Yield: 5.5 g (83 % of theory )

pp: 149 - 150 ⁰ C 1. AOOH) EPEW §0 -89.9 ° (o « in BAE, CU, DC uniform

1.3. Boc-Tyr-Pro-OH

2.3 g Böő-Tyr-OTOP were dissolved in 10 mL of THP and added at 0 ⁰ C with 0.8 g of proline, 0.4 g HOBt and 0.98 ml TSA. The mixture was stirred at 0 ^° C. for 1 h and at RT for 3 h. Thereafter, the LM i.Vak. evaporated and the oily residue taken up in saturated NaHCOy solution. NaOH three times the extraction of the aqueous phase with EE this was brewed with citric acid to pH and then with several times. EE extracted. The combined EA phases were washed neutral with water over Na ₂ SO ₄ . dried and the LM evaporated. Recrystallization from EE yielded the DC uniform product in BAE, CM and EPEW

Yield: 1.4 g (74 % of theory )

Pp: 116-118 ⁰ C

^ ² D ⁼ ~ ³³ » ⁵ ° ^{(about 1} » ^Ac0H) 1.4. Boo-Tyr-Pro-D-Phe-Pro-Gly-OH

To prepare the mixed anhydride, 1.22 g of Boc-Tyr-Pro-OH, 40 μl of NEM and 418.6 μl of CAIBE were added in 25 ml of THP

at -15 ⁰ O together and gave the Reaktlonsgemisoh calcined with 1.58 g of HD-Phe-Pro-Gly-ONB · HCl (obtained by Entaoylierung of 1.2 with 1.1 N HCl / AoOH -Pp: 190 ⁰ C) (Zers.), ^ J ²⁰ J " -123 _f 8 ° (o-1, AoOH)) and

NaOH a reaction time of 1.5 h at -15 ⁰ C and 5 h at RT, the reaction mixture was worked up in the usual manner and the crude product was reprecipitated twice from EE ether · In BAE, CM and EPEW DC unitliohe The product (2.2 g = 84 % of theory , / f ²⁰ J = -74.4 ° (o = 1, AoOH) was dissolved in 60 ml of MeOH and 1 ml of water and added with 200 mg of Pd- After 3 h of reaction time, the catalyst was filtered off and washed with MeOH.The LM was evaporated off in vacuo and the oily residue was reprecipitated from EA ether and isopropanol-diisopropyl ether for further purification g.) over a silica gel G-60 (Merck) column (eluent CHCIv-MeOH 8: 2). Chromatograph! ert. Further accidents from EE ether afforded the product amorphous in BAE, CM and EPEW DC unitlio

Yield: 1.2 g (. 55% of theory). Pp: 210-217 ⁰ C (dec.)

20 = -78.2 ° (ο = 1, AcOH) D

1.5. H-Tyr-Pro-D-Phe-Pro-Gly-OH · HCl

1.2 g of 1.4 × were dissolved in 15 ml of dioxane and combined with 5 ml of 6N HCl / dioxane. The product was precipitated with ether for a further 30 minutes at RT, filtered off and washed with ether. Accidents of MeOH ether were reported in the BAW, BSWE and BPEW DC-uniform product

Yield: 1.0 g (92 % of theory )

Pp: 222-227 ⁰ C (dec.) -47.1 ° β 0 (o = 1, AOOH)

Variant II: step-by-step condensation II.1. Boo-Pro-Gly-OMe

5.38 g of Boo-Pro-OH, 3.175 ml of NEM and 3.25 ml of CAIBE in 50 ml of THP were reacted with 3.14 g of H-Gly-OMe.HCl and 3.175 ml of NEM and worked in analogy to the procedure for 1.1 in the same way. The crude product was recrystallized from EE-PE and from diisopropyl ether.

Yield: 5.0 g (70 % d · Th ·)

Pp: 70.5 - 72 ⁰ C

/ KJ ² J ^a -78.5 ° (c = 1, AcOH)

DC uniform in BAE, CM and EPEW

II.2. Boc-D-Phe-Pro-Gly-OMe

4.6 g of Boo-D-Phe-OH, 2.2 ml of NEM and 2.25 ml of CAIBE in 40 ml of THP were mixed with 3.85 g of H-Pro-Gly-OMe.HCl (obtained by deacylation of II x 1 · With 1.1 N HCl / AcOH oil, 0 0 ² p = -57.7 ° (c = 1, AcOH)) and 2.2 ml NEM as under 1.2. The reaction was carried out analogously and the product recrystallized from EE diisopropyl ether.

Yield: 5.25 g (70 % of theory )

Pp: 138-139 ⁰ C

βCj ² g = -102.5 ° (c = 1, AcOH)

DC uniform in BAE, CM, E & EW

II.3 · Boc-Pro-D-Phe-Pro-Gly-OMe

1.64 g Boo-Pro-OH was dissolved at -15 ⁰ C in 20 mL of THP and treated with 0.97 ml of NEM and 0.99 ml CAIBE. After 10 minutes, 2.82 g of HD-Phe-Pro-Gly-OMe.HCl (obtained by Esscacy) were added.

lation of IX. 2. with 1.1 N HCl / AcOH - JPp: 128-131 ⁰ C, JU ² S * -138.8 ° (ο β 1, AoOH)) and 0.97 ml NBM for reaction

tiong and left for 1 h at -15 ⁰ C and 4 h at RT. The batch was worked up as usual and the crude product of EE-PE was reprecipitated.

Yield: 3.2 - <79 % d. Th.) Pp: 65-71 ⁰ O (dec.) Gtj ²⁰ J = -117.9 ° (c = 1, AcOH) PC uniform in BAB, CM, BPEW

II. 4. "Boo-Tyr-Pro-D-Phe-Pro-Gly-OMe

920 mg of Boo-Tyr-OToP were admixed with 980 mg of H-Pro-D-Phe-Pro-Gly-OMe.HCl (obtained by deacylation of II.3. Using 1.1 N HCl / AcOH - Pp: 127-130 ⁰ C, EQ ² J = -48.2 ° (o = 1, AcOH)), 270 mg of HOBt and 3.2 ml of TEA in 20 ml of THP 1 h at 0 ⁰ C and allowed to react at RT for 12 h. Bs was worked up as usual and the crude product was reprecipitated from BE-PB.

Yield: 1.1 g (. 79% of theory). Pp: 113-120 ⁰ C (dec.) BQ ² I = -82.6 ° (ο β 1, AOOH) DC uniformly in BAE, OM, BPEW

II.5. 3oC-Tyr-Pro-D-Phe-Pro-Gly-0H

980 mg II "4 * were dissolved in 25 ml of acetone and reacted with 2 ml of 1 N NaOH. After a reaction time of 5 h at RT. wu.vde the organic LM i.Vak. evaporated and the residue gellist in 15 ml of saturated NaHCOv solution. The aqueous phase was extracted twice with BB and then acidified to pH 3 with solid citric acid. After three extractions with EB, the combined ones were

ϊ 7 22 27

JiE phases mi ⁴ ; Water until neutral, dried over Na ₃ SO ₄ and concentrated by evaporation in vacuo. evaporated · The product was reprecipitated from MeOH / ether and then chromatographed over a silica gel G 60 (Mevck) column (eluent CHCl ₃ -MeOH 8: 2). Repeated seizure from EE / ether yielded 625 mg (65 % of theory ) of the product which was unitary in BAE, CM and EPEW DC (analytical data corresponds to the maintenance indicated under 1.4).

II.6. H-Tyr-Pro-D-Phe-Pro-Gly-OH · HCl

600 mg II.5. were dissolved in 10 ml of dioxane and treated with 3 ml of 6N HCl / dioxane. Furthermore, as under 1.5. described procedure. This resulted in 515 mg (95 % of theory ) of the BAE ₁ BEWE and BPEW DC-uniform product, which in its analytical data was less than 1.5. the specified values.

Example 2:

Application of (D-Phe ^) - β-Casomorphine-5 (Tyr-Pro-D-Phe-Pro-Gly) in Anhydrous Solution to Isolated Atrial Auricles scrie isolated perfused sea-hearts hearts. Experiments were performed on isolated auricles and isolated perfused hearts performed by guinea pigs. The preparations were investigated in Og-saturated Tyrode Iösung at 30 ⁰ C. The isometric contractions were measured with strain gauges and the coronary flow by electrical drop counting. The product of contraction amplitude and frequency gave an index of the performance of the auricular preparations. The evaluation of the arrhythmia was carried out according to a scheme in which the degree of grading of the individual arrhythmia forms was divided into levels between 0 and 100. The evaluation of the effects was expressed as a percentage change compared to the corresponding controls. The results were obtained by Student'st test

checked for significant differences · The investigated peptide showed positive inotropic, coronary dilatatory and antiarrhytmic effects (Tables 1-3) ·

Example 3:

On the isolated atrial-auricular preparation and isolated perfused hearts of the guinea pig, the positive influence of isoprenaline on contraction force, heart rate and coronary flow was significantly reduced by casomorphine-type peptides (Table 4).

Example 4:

Casomorphine-type peptides modulate the binding of the cardiac glycoside ouabain to cardiac membranes in a concentration-dependent manner. Low-peptide concentrations (10.sup.-10 to 10.sup.-1 mol / l) significantly increase the .alpha.-ouabain binding as a result of high .beta.-casomo.prin concentrations (10 to 10 "mol / l) significantly lowered (control: <H-ouabain binding without peptide addition = 100 %) , (Table 5)

Example 5:

Effect of casomorphin-type peptides on cardiovascular parameters in vivo:

Casomorphine-type peptides cause a significant decrease in heart rate to 90 pounds up to 40 % over control (100 %) and a reduction in mean arterial blood pressure to 90 % to 80 % (control 100 %). In all cases, the initial value (control value) can be measured again 1 to 2 minutes after the application. · While the casomorphine peptides influence the heart rate more than the mean arterial blood pressure, the opposite effects occur with the adipose substances bradykinin and angiotensin II (Table 6). ·

T a b e 1 1 e 1

Effect of Tyr-ro-D-Phe-Pro-Gly on the cardiac output of isolated guinea-pig atrial auricular preparations (cardiac output as the product of contraction amplitude and heart rate) (n = number of experiments)

Concentration increase

(mol / 1) contraction beat rate cardiac output

amplitude (cm) (SchlSge / min) effect (.%)

10 ~ ⁹ 8 1.2 + 0.05 168 + 3.9 20 + 6.0

10 ~ ⁸ 8 1.0 + 0.04 170 + 8.9 36 + 6.1

10 " ⁷ 7 1.1 + 0.05 158 + 7.6 22 + 5.3

Table 2

Effects of lyr-Ero-D-Phe-Ero-Cly on contraction force and coronary flow isolated

Konz · η contraction amplitude Effect 06) coronary flow increase IO «« * (Mol / 1) control 22 - 6,8 control Effect (56) ls> (cm) 30 ± 4.5 (Ml / min) 30 ± 4.1 IO 10-9 8th 2.5 i 0.07 15 ^± 5.1 6.2 ± 0.2 32 ± 4.5 IO ΙΟ " ⁸ 8th 2.6 i 0.05 7.1 ^± 0.5 26 ± 3.7 "» 4 10-7 8th 2.8 ± 0.05 8.4 ^± 0.4

Table 3

Influence of Tyr-Pro-D-Phe-Pro-Gljr on aconitin-induced arrhythmia of isolated spontaneously beating guinea-pig auricular preparations (n = number of experiments)

Conc. (Mol / 1)

Improvement (%) of aconitine-induced arrhythmia

-9

10- " ⁸ 10-7

22 ± 5.4 42 ± 7.2 18 ± 6.6

Table 4

Effect of isoprenaline on isolated atrial-auricular preparations under the influence of fever casomorphine-type (n = Ατικηνι the experiments)

Casomorphin peptides

Tyr-D-Pro-Phe-Pro-Gly Tyr-Pro-D-Phe-Pro-Gly Lyr-Pro-Tyr-Pro-Gly

lyr-Pro-Phe-Pro-Gly (β-Casomorphine-5)

Conc. (Mol / 1) inhibition (%)

Cardiac Output Under Isoprenaline (1.6 χ 10 ~ ⁸ mol / l) (8 χ 10 ~ ⁸ mol / l)

10 10 10 10

-8th

-8th

-8 ± 5.6 i 5.1

± 8.3

± 4.3

82 ± 9.4 52 ± 7.7 35 ^± 8.4 50 ± 6.3

table

Influence of casomorphine-type peptides on the ^ H-ouabain binding to cardiac guinea pig heart membranes (n = number of experiments in double puncture, s = significant, ns = not significant)

Casomorphin peptides

Concentration (mol / 1)

Tyr-Pro-Phe-Pro-Gly 5 (β-Casomorphine-5)

Tyr-Pro-Phe-Pro 4

Pro-Phe ~ Pro-Gly 4

Tyr-Pro-Phe-D-Pro-Gly 6

Tyr-Pro-D-Phe-Pro-Gly 4

% Binding

% -Ouabainkonz.

(Control 100 %)

Concentration % binding

(mol / 1) ³ H-ouabain conc.

(Control 100 %)

10

-8th

10

-8th

10

-8th

10

-8th

10

-8th

i 21 s. 10

i 6 ns. 10

- 22 s. 10

- 10 s ·

± 9.5 ns. 10

-5

, -5

, -5

10

-5

-5

, B ± 5.4

108 , 0 i 5.3 s. ro 96 , 0 ± 9.1 ns. ro 111 t0 ± 7.6 s. ro IO 80 , 2 ± 14.4 s.

Table 6

Effect of peptides of the casomorphin-type on heart> circulatory parameters in vivo on non-anesthetized Batten (i.v. administration: 0.5 mg peptide / kg body weight, η = number of experiments)

Casomorphin peptides

Masinial decrease duration until the heart rate reaches the % baseline value

(Control: 100 %) in s Maximum duration of time until the mid-point is reached. The lower arterial output value is reached. Blood pressure in s on % (control: 100 %)

Tyr-Pro-D-Phe-Pro-Gly 6

Tyr-Pro-Phe-Pro-Gly 6

Tyr-Pro-Phe-D-Pro-Gly 6

Tyr-D-Ala-Phe-D-Pro-Gly 6

81 93 86

37

60

60

60

210 90 105 117

10

10

20

210

bradykinin

angiotensin

113 86

120 12068 130

180 120

100 ng / kg

ro ο

Claims (5)

1. A process for preparing cardiopulmonary preparations, characterized in that as an effective component oasomorphine-type peptides and their derivatives in the low concentration range of 1O " ⁹ to 10"'mol / 1 and 1 ug / kg to 1 mg / kg be administered. 2. An ancestor according to claim 1, characterized in that as active components linear, cyclic or oyclischverzweigte peptide analogues of the gasomorphine type, their salts or C- and / or N-terminal modified derivatives, preferably with primary or secondary aliphatic, aromatic or heterocyclisoher C-terminal amide structure can be used, wherein proline in one or both positions by hydroxyproline, dehydroproline, homoproline, pipecolic acid, alanine, phenylalanine, valine, leucine, isoleucine, lysine, ornithine, 2,4-diaminobutyric acid, arginine, cyctein, methionine , Serine, threonine and phenylalanine may be replaced by homophenylalanine, N-methylphenylalanine, tyrosine, N-methyltyrosine, the corresponding ring-substituted compounds, tryptophan, thyronine, thyroxine, valine, leucine, isoleucine and alanine. The amino acids can be contained in the L- or D-configuration or as a racemate in the peptide structures. Glycine may or may not be present. If present, it can be replaced by any amino acid in the L or D configuration or as a racemate. In addition, the corresponding N-terminal tripeptides of the above-mentioned basic structure and their derivatives can also be used as the active component. Some preferred compounds are, for example, the oligopeptides Tyr-Pro-D-Phe-Pro-Gly, Tyr-D-Ala-Phe-D-Pro-Gly, Tyr-Pro-Phe-Phe-Gly-NHg, Tyr-Pro-VaI -Pro-Gly, Tyr-Pro-D-Val-Pro-Gly and Tyr-D-Met-Phe-Pro-NHg. 3. The method according to claim 1 and 2, characterized in that the peptides of the casomorphine type by chemosynthesis Segmentkondendation or stepwise extension of the peptide chain from the C-terminal end - according to the usual methods in peptide chemistry or by methods of enzyme-catalyzed peptide synthesis or by combination Both synthesis variants are produced · 4. Process according to Claims 1 to 31, characterized in that improved therapeutics are produced on the basis of known cardiotonic and antianginal and antiarrhythmic drugs with addition of 0.1 to 1.0 % of the casomorphine-type peptides and their derivatives. 5. The method according to claim 1 and 2, characterized in that the adrenergic effect is reduced under the influence of the casomorphine-type peptides and their derivatives.