||||||III US005565423A United States Patent (19 11 Patent Number: 5,565,423 Sandow et al. 45) Date of Patent: Oct. 15, 1996

(54) CYCLOPEPTIDES AND THEIR USE AS ABSORPTION PROMOTERS WHEN APPLIED TO THE MUCOSA 75) Inventors: Jirgen Sandow, Glashitten/Taunus; Walter Dirckheimer, Hattersheim am Main; Ginter Ditzinger, Frankfurt am Main, all of Germany; Hans-Peter Merkle, Zürich, Switzerland (73 Assignee: Hoechst Aktiengesellschaft, Frankfurt am Main, Germany 21 Appl. No.: 418,882 (22 Filed: Apr. 7, 1995 Related U.S. Application Data or a physiologically tolerated salt thereof, (I) 63) Continuation of Ser. No. 65,013, May 24, 1993, abandoned. in which X is a basic L- or D-amino acid, 30 Foreign Application Priority Data Y is a neutral or basic L- or D-amino acid, May 26, 1992 (DE) Germany ...... 42 17 350.7 Z is a neutral or basic L- or D-amino acid, which can be identical to or different from Y, (51) Int. Cl...... A61K 38/12 I52) U.S. Cl...... 514/11; 514/2; 514/9; R" is an N-bonded acyl radical of the formula II 530/317 Ar-(CH2) CO-, (II) 58 Field of Search ...... 514/2, 9, 11; 530/317, N 530/318, 319, 320 N 56 References Cited OCH3 U.S. PATENT DOCUMENTS in which Ar is a phenyl radical, which is optionally substi tuted by 1, 2 or 3 identical or different radicals from the 3,817,973 6/1974 Bouchaudon et al...... 530/319 series comprising hydroxyl, (C-C)-alkoxy, amino, car 5,091,365 2/1992 Sandow et al...... 514/9 boxyl, (C-C)-alkylamino and halogen, or is a 2-aminothia 5,177,059 1/1993 Handley et al...... 530/317 zol-4-yl radical, FOREIGN PATENT DOCUMENTS p is an integer from zero to 4 and q, n,m and 1 independently 0302466 2/1989 European Pat. Off.. 2204887 9/1972 Germany. of one another are Zero or 1. 0005709 2/1971 Japan ...... 530/317 The invention additionally relates to the use of compounds Primary Examiner-Howard E. Schain of the formula I and physiologically tolerated salts thereof Attorney, Agent, or Firm-Finnegan, Henderson, Farabow, for promoting the absorption of peptides and proteins when Garrett & Dunner, L.L.P. applied to the mucosa, and pharmaceutical formulations which contain a pharmacologically active amount of one or 57 ABSTRACT more peptides or proteins and a compound of the formula I Cyclopeptides and their use as absorption promoters when or physiologically tolerated salts thereof. applied to the mucosa The invention relates to a compound of the formula I 8 Claims, No Drawings 5,565,423 1. 2 CYCLOPEPTDES AND THEIR USE AS in which X is a basic L- or D-amino acid, ABSORPTION PROMOTERS WHEN Y is a neutral or basic L- or D-amino acid, APPLIED TO THE MUCOSA Z is a neutral or basic L- or D-amino acid, which can be This application is a continuation of application Ser. No. 5 identical to or different from Y, 08/065,013, filed May 24, 1993, now abandoned. R is an N-bonded acyl radical of the formula II (II) DESCRIPTION A-CHy-Fi CO-, 10 N The invention relates to cyclopeptides and their use for N promoting the absorption of peptides and proteins when OCH; ), applied to the mucosa. The use of peptides and proteins as medicaments is made in which Ar is a phenyl radical, which is optionally substi considerably more difficult by the problems of a suitable tuted by 1, 2 or 3 identical or different radicals from the pharmaceutical formulation from which the peptide or pro 15 series comprising hydroxyl, (C-C)-alkoxy, amino, car tein to be used therapeutically or diagnostically is absorbed boxyl, (C-C)-alkylamino and halogen, or is a 2-aminothia in an adequate amount and reliably. zol-4-yl radical, p is an integer from Zero to 4 and q, n, m The administration of one or more daily individual doses and 1 independently of one another are Zero or 1. by nasal application, either in the form of nose drops or by 20 spraying a suitable solution into the nose, is known J. Those radicals X, Y and Z which are derived from Sandow, W. Petri (1985), Transnasal Systemic Medications, naturally occurring amino acids (cf., for example, Schr Verlag Elsevier, 183-199). Readily tolerated aqueous solu 6der, Libke, The Peptides, Volume 1, New York 1965) and tions with added preservatives and if appropriate auxiliaries 2,4-diaminobutyric acid (Dab), antipodes thereof and simple to increase the absorption are used for this. The customary 25 auxiliaries for increasing absorption (absorption enhancers) metabolites, which, if they are chiral, can be present in the all irritate the mucosa or are unsuitable due to an unpleasant D- or L-form, are preferred. Y-Dab is 2,4-diaminobutyric smell or taste, and often already lead to considerable pain acid linked to the ring via the Y-amino group. and lacrimation after a single application, or generate a Unless stated otherwise, the three-letter symbols (cf., for progressive irritation and inflammation of the nasal mucosa 30 example, Pure Appl. Chem. 56 (1984) 595-624 and Eur. J. after several applications. This applies, for example, to derivatives of fusidic acid, to bile acids, to surfactants and to Biochen. 138 (1984) 9-37) for the radicals of the amino the various glycols (polyethylene glycol, polypropylene acids are used. These symbols are preceded by the symbol glycol). The use of cyclopeptides based on naturally occur "D' if the radical is the radical of a D-amino acid; radicals ring substances for promotion of the absorption of peptides 35 without a configuration symbol have the L configuration. and proteins on application to the mucosa is furthermore The invention relates both to the optically pure com described in EP-B-0 302 466. These cyclopeptides are a mixture of chemically similar compounds; they must there pounds and to stereoisomer mixtures, such as enantiomer fore be purified for pharmaceutical use. Above all, they have mixtures and diastereomer mixtures. an intrinsic antibiotic activity, which may lead to the devel 40 Compounds of the formula I which are preferably opment of resistance in certain germs. employed are those in which The object is therefore to provide compounds which X is lysine, ornithine, 2,4-diaminobutyric acid or arginine, improve the spectrum of action of peptides and proteins Y and Z independently of one another are lysine, orni employed as pharmaceuticals by effecting, with good toler thine, 2,4-diaminobutyric acid, arginine, threonine or serine ability and lack of antibiotic partial action, good promotion 45 and of absorption. R', n,m and 1 have the abovementioned meaning, it being According to the invention, this object is achieved by possible for the amino acids each to be present in the D- or compounds of the formula I L-form. 50 The following compounds are especially preferred: - Dab -G Leu (I) Peptide PMB-1:

Dab D-Phe Dab - G Leu

55 / ND-Phe Thr Dab

Thr Dab tipal 60 Xn /co N Y-Dab N Yn N Z 65 Dab-CO-CA W Ri, S ^s, and physiologically tolerated salts thereof, 5,565,423 4 Peptide PMB-II: concentration through vaginal, rectal or buccal medicament forms (i.e., for example, films, tablets or suppositories) also leads to no irritation of the mucosa. For most of the peptides and proteins known today which are used or will shortly be used as therapeutics or diagnos tics, application to the mucosa, such as, for example, nasal, buccal, rectal, vaginal or pulmonal use, but in particular nasal, is appropriate and possible. Thr Dab Peptides and proteins which are suitable for this are those 10 which comprise 3 to 225 amino acids, such as, for example, TRH (protirelin, thyroliberin), LHRH (gonadoliberin), N 4 OH chemically modified peptide analogs of hypothalamic regu D latory hormones, such as, for example, buserelin, somatosta Dab-CO OH tin and cyclic somatostatin analogs, somatorelin, GRH ana 15 logs, peptide analogs of hypophysis hormones, such as, for example, the corticotropin analog alsactide (ACTH-17), OH calcium-regulating hormones (calcitonin, parathormone) and their analogs, as well as gastrointestinal hormones (for Peptide PMB-III: example secretin and cholecystokinin) and pancreatic hor 20 mones (insulin and insulin analogs). Those having 3 to 51 amino acids are particularly suitable. / Dab - Se N The following may be mentioned in particular:

Name of the Number of Dab - 25 peptide or protein Use, for example for: amino acids Oxytocin uterine inertia 9 Vasopressin diabetes insipidus 9 Ornipressin hemorrhages 9 r Dab Desmopressin diabetes insipidus 9 30 Corticotripin inflammatory 39 7D) (ACTH) diseases Dab-CO-CH2 Tetracosactide inflammatory 24 diseases Alsactide 17 Insulin diabetes mellitus 51 The compounds according to the invention are prepared, 35 beta-sleep ind. Peptide sleep disturbances 9 for example, using the general methods of peptide chemistry Secretin gastric hemorrhages 27 (Houben-Weyl, Methoden der organischen Chemie (Meth Cholecystokinin diseases of the 8-32 ods of organic chemistry), 15/1 and 2). The compounds can biliary tract, as an be prepared, for example, in stages starting from polymyxin appetite suppressant 40 Somatoliberin retarded growth 44 B heptapeptide (formula I:1=m=n=0; R'=H), or from poly (GRH) myxin B octapeptide (formula I:1=m=0; X=L-Dab; R'=H) (D-Ala2Somato- 29 or from polymyxin B nonapeptide (formula I:1=0, Y-L- liberin-(1-29)- Thr; X-L-Dab; R=H), the amino groups of which, with the amide Somatoliberinyl- 45 exception of that to be acylated, are protected, and the glycine corresponding activated carboxylic acids (for example car 45 Glucagon hypoglycemia 29 boxylic acid halides, mixed anhydrides or active esters). Somatostatin gastric hemorrhages 14 The starting peptides can be obtained by reductive cleav Octreotide tuOS 8 age of N-Boc-protected polymyxin B, for example using Spantide substance Pinhibition 1. Corticoliberin hypophysis diagnostic 41 metal hydrides, for example NaBH or NaCNBH, or by (CRF) enzymatic cleavage, for example with the protease ficin, 50 Bradykinin allergic and inflam- 9-11 which is known from the literature. In this case, the N-oxido antagonists matory disorders pyridyl-methoxycarbonyl protective group has proved to be Atriopeptin III cardiac and renal 24 insufficiency a suitable N-protective group. ANF-(99-126) 28 The invention furthermore relates to the use of com Thymopentin rheumatoid arthritis 5 pounds of the formula I for promoting the absorption of 55 Interferon-alpha colds 125 peptides and proteins on application to the mucosa. The Thyroliberin hypophysis diagnostic 3 compounds according to the invention help quite consider (TRH) ably to improve the absorption of peptides and proteins on Gonadoliberin cryptorchism, sterility 10 application to the mucosa. The increase in the activity (based Buserelin prostate cancer, 9 on improved absorption) of the peptides or proteins after 60 endometriosis Goserelin prostate cancer, 0 addition of the compounds according to the invention is thus endometriosis 300 to 400%, and can be more than 1000% in an individual Triptorelin prostate cancer, 10 C2SC. endometriosis The compounds according to the invention moreover LH-RH-T prostate cancer, 9 endometriosis cause no pain sensation and damage to the mucosa in 65 Leuprorelin prostate cancer, 9 humans following nasal application of 1 to 200 ul of a endometriosis concentration of 10 to 10 mol/l. Local use of the same 5,565,423 5 6 -continued pulmonarily or vaginally. Nasal application is preferred here. Name of the Number of The pharmaceutical formulations according to the inven peptide or protein Use, for example for: amino acids tion can be prepared by the method known to the person Lutrelin prostate cancer, 9 skilled in the art with addition of excipients suitable for the endometriosis production of pharmaceutical preparations. Pharmaceuticals Nafarelin prostate cancer, 10 endometriosis are especially suitable for application to the mucosa, such as, Histrelin prostate cancer, 9 for example, tablets, suppositories, capsules, gels, films, endometriosis emulsions, suspensions, aerosols, solutions or sprays Calcitonin Paget's disease, 32 (Sucker, Fuchs, Speiser, Pharmazeutische Technologie osteoporosis O (Pharmaceutical Technology), Georg Thieme Verlag 1978). Elcatonin Paget's disease, 32 The following are preferably used: osteoporosis Parathormone-(1-34) hypocalcemia 34 1. Aqueous or aqueous-alcoholic solutions for application Sincalide diagnostic for 8 with a dropper pipette or a plastic squeeze bottle, or for pancreatic function nebulization with a metered nebulizer pump. 15 Ceruletide r 10 In addition to the active compound and the absorption Pentagastrin diagnostic for gastric 5 promoter, the formulation can for example comprise an function Desglugastrin 7 isotonicizing additive, for example sodium chloride, potas Ocitide intestine-contracting 5 sium nitrate or potassium sodium phosphate, polyalcohols, Angiogenin tissue regeneration 123 such as, for example, glucose, mannitol or sorbitol, buffer TGF-beta tumor therapy, 224 substances, such as, for example, potassium sodium phos immunosuppression phate, citric acid and its salts and mixtures of the two, in Hirudine inhibition of 64, 65 coagulation order to establish a pH range from 3 to 8, a preservative, for GM-CSF stimulation of bone 27 example benzalkonium chloride, benzyl alcohol, 1,1,1- aOW trichloro-2-methyl-2-propanol or methyl 4-hydroxyben Erythropoietin antianemic agent 165 25 zoate, a chelating agent, for example sodium EDTA, and, as the solvent, water or mixtures of water with (C-C)- These peptides and proteins can be obtained by generally alkanols. The solution is applied with a suitable apparatus or known processes, for example by Merrifield synthesis, sprayed into the nose or onto the oral mucosa. genetic engineering and by isolation of naturally occurring 2. Aqueous or aqueous-alcoholic gels for introduction into peptides and proteins. 30 body cavities (mouth, nose, rectum, vagina) The invention furthermore relates to pharmaceutical for In addition to 1., a gel comprises an additive which mulations comprising increases the viscosity, for example a polyacrylate polymer a) a pharmacologically active amount of one, two or three or a cellulose ether, such as, for example, hydroxypropyl peptides or proteins in each case comprising 3 to 225 amino methylcellulose (HPMC), hydroxyethylcellulose (HEC) or methylhydroxyethylcellulose (MHEC). acids, in particular 3 to 51 amino acids or physiologically 35 tolerated salts thereof, and 3. Suspensions in propellent gases b) at least one auxiliary of the formula I. In addition to the micronized active compound and the Preferred formulations are those which comprise an aux micronized absorption promoter, the formulation can com iliary of the formula I in which prise, as a propellent gas, a chlorofluorohydrocarbon, for X is lysine, ornithine, 2,4-diaminobutyric acid (Dab) or example “Frigen F 11, 12 or 114 (R)=a registered trademark arginine, 40 of Hoechst AG), or mixtures thereof or a fluorohydrocarbon Y and Z independently of one another are lysine, orni such as tetrafluoroethane (HFA 134a) or heptafluoropropane thine, 2,4-diaminobutyric acid, arginine, threonine or serine (HFA227) and a suspending auxiliary, for example sorbitan and trioleate, oleic acid or lecithin. The containers are filled in a R', n,m and 1 have the abovementioned meaning, it being manner which is known perse by the low temperature filling possible for the amino acids each to be present in the D- or 45 process, or by filling under pressure. L-form. 4. Triturations with carrier auxiliaries filled into capsules Especially preferred formulations are those which con for intranasal or inhalative use prise at least one auxiliary chosen from the peptides PMB-I Hard gelatin capsules are filled with the micronized to -III. substances (active compound and absorption promoter), if The pharmaceutical formulations according to the inven 50 appropriate after addition of an agent for improving the flow tion furthermore preferably comprise a peptide or protein properties, such as lactose. The contents of one capsule is comprising 3 to 225 amino acids, in particular a peptide or applied intranasally or pulmonarily with an inhalation aid protein comprising 3 to 51 amino acids. which enables the powder to be converted into inhalable However, formulations containing two or three different fumes. peptides and/or proteins, such as, for example, corticotro 55 5. Buccal forms pin-LHRH+GRH, or protirelin+LHRH+GRH, in combina The active compound and absorption promoter can be tion with at least one auxiliary of the formula I, are also of present in dissolved or suspended form. Compressed articles interest, in particular for use as a diagnostic. or laminates of mixtures of the active compound and absorp The dose of the peptides and/or proteins and of the tion promoter in polymers are suitable drug forms. Possible auxiliaries of the formula I in the formulations or products 60 according to the invention when used on mammals, prefer polymers are cellulose ethers (for example HPMC or car ably on humans, is in the range from 10 ug to 10 mg per boxymethylcellulose (CMC)) or polyacrylates. peptide?protein and use, and, for the auxiliary of the formula EXAMPLE I, is at a concentration of 10 to 10 mol/l per use, Synthesis of PMB-I preferably between 10 and 10 mol/l. 65 The formulations according to the invention can be used 1.26 g (1 mmol) of "tetra-Boc-polymyxin B-octapeptide' by application to the mucosa, i.e. nasally, buccally, rectally, are dissolved in 10 ml of pyridine, and 0.318g of the HOBt 5,565,423 7 8 ester (HOBt=1-hydroxybenzotriazole) of 2-amino-thiazol After a reaction time of 20 hours at room temperature, the 4-yl-2-methoximinoacetic acid is added. After 24 hours, the residue is dissolved in 200 ml of ethyl acetate, and the pyridine is evaporated in vacuo, the residue is dissolved in solution is washed with 200 ml of N/10 hydrochloric acid 200 ml of ethyl acetate and the solution is washed first with and with 200 ml of water. After drying over sodium sulfate, 200 ml of N/10 hydrochloric acid and then with 200 ml of the solvent is removed, the residue is dissolved again in a water. After drying over sodium sulfate, the solvent is mixture of ethyl acetate/methanol (85:15), and the solution removed, the residue is dissolved again in a mixture of ethyl is chromatographed over a silica gel column (4x50 cm; acetate/methanol 85:15, and the solution is chromato 70-200 um; 200 g) using the same solvent mixture. 40 graphed with the same solvent mixture over a silica gel fractions of 15 ml are removed. Fractions 20–39 contain the column (4x50 cm; 70-200 um; 200 g). 40 fractions of 10 ml 10 desired product (CHNOs). The solventis removed in are collected. Fractions 21-35 contain the desired substance. vacuo on a rotary evaporator. The solvent is removed in vacuo on a rotary evaporator. Yield of the intermediate:0.85 g (61.2%) Yield of the intermediate: 1.1 g (76% of theory) Molecular weight:1386 g/mol (Li salt) Molecular weight:1445 g/mol (CHNOS) 15 R value (thin layer chromatography as Example 1)=0.57 R value (thin layer chromatography silica gel 60 F Splitting off the protective groups was carried out as in 254/Merck, Darmstadt, ethyl acetate/methanol 85:15)=0.51 Example 1. Splitting off the Boc protective groups: Yield:0.59 g (97.7%) 1.1 g of the intermediate are dissolved in 10 ml of Molecular weight:980 g/mol (C7H7N Oo). trifluoroacetic acid, while cooling with ice, and the solution 20 is then left to stand at room temperature for 2 hours. After R value (thin layer chromatography solvent mixture as in concentration, the solid residue is triturated with ether, Example 1): 0.26 filtered off, rinsed with ether, and dried over POs in vacuo. EXAMPLE 4 Yield:0.96 g (78.2%) Molecular weight:1046 g/mol 25 Investigation of the action of LHRH or LHRH CF,COH) analogs R value (thin layer chromatography silica gel 60 F A method which is suitable for demonstrating the inten 254/Merck, Darmstadt, mobile phase n-butanol (50), pyri sification of action of a peptide or protein by means of a dine (20), glacial acetic acid (6), water (24)=0.11. 30 cyclopeptide according to the invention is, for example, determination of the release of LH in male rats (100 g body EXAMPLE 2 weight) anesthetized by ethyl carbamate. In this method, the hormone release is compared over a period of 6-7 hours Synthesis of PMB-II after treatment, i.e. after nasal or rectal application of LHRH 1.25 g (1 mmol) of “tetra-Boc-polymyxin B octapeptide' 35 or LHRH analogs in physiological saline solution with or are dissolved in 10 ml of pyridine, and 0.287 g of the HOBt without addition of cyclopeptides of the formula I. It was ester of gallic acid is added. After 48 hours, the pyridine is found that, for example, a 21- to 32-fold increase in the stripped off, the residue is dissolved in 200 ml of ethyl action of a nasally administered dose of 10 ng buserelin is acetate and this solution is washed first with 200 ml of N/10 achieved by the peptides PMB-I to -III in a concentration of hydrochloric acid and then with 200 ml of water. After 40 0.01 M, based on the release of LH. The action of a nasally drying over sodium sulfate, the solvent is removed, the administered dose of 800 ng of LHRH was intensified by a residue is dissolved again in a mixture of ethyl acetate/ factor of 3 to 6 by the peptides PMB-I to -III in the same methanol (85:15), and the solution is chromatographed with animal model, also based on the areas under the curves. the same solvent mixture. Silica gel (70-200 um), which has EXAMPLE 5 been washed beforehand with half-concentrated hydrochlo 45 ric acid to remove iron, and then with water until a neutral Investigation of the Intensification of Action of reaction is obtained, and dried at 130 C., is used as the ACTH carrier material. 40 fractions of 15 ml are eluted. Fractions 34-39 contain the desired substance. The solventis removed The action of cyclopeptides of the formula I on the in vacuo on a rotary evaporator. 50 absorption of ACTH (corticotropin) and ACTH analogs was investigated, after application of the test substances accord Yield of the intermediate:0.89 g (62.8%) ing to Example 4, on male rats (100 g body weight) Molecular weight=1414.58 g/mol. anesthetized by phenobarbital or ether. The release of cor Splitting off the Boc protective groups was carried out ticosterone was determined in the serum by specific radio analogously to Example 1. 55 immunoassay as the parameter of the action. It was found Yield:0.79 g (86%) that after nasal treatment, for example, with the ACTH Molecular weight:1014 analog alsactide (ACTH-17) in a dose of 1.5 ug in the presence of peptides (PMB-I to -III, the corticosterone 4CFCOH-1470.21) release was increased 2- to 3-fold over 3 hours. R value (separation system as under Example 1):005. 60 EXAMPLE 6 EXAMPLE 3 Investigation of the Intensification of Action of Synthesis of PMB-III Calcitonin 1.26 g (1 mmol) of "tetra-Boc-polymyxin B octapeptide" 65 The action of cyclopeptides of the formula I on the are dissolved in 10 ml of pyridine, and 0.233 g (1 mmol) of absorption of calcitonin and calcitonin analogs (for example the hydroxysuccinimide ester of phenylacetic acid is added. salmon calcitonin) was investigated on male rats of 100 or 5,565,423 10 200 g body weight after application according to Example 4, EXAMPLE 10 by determination of the serum calcium level over a period of 1-6 hours after the treatment. It was found that the action of a nasal dose of 1.2 g of Ge. salmon calcitonin was increased 3.4-fold by addition, for example, of peptide PMB-I in a concentration of 0.01 M, based on the decrease in the serum calcium level. Alsactide (ACTH-17) 0.003 mg Peptide PMB-II 1.200 mg Polyacrylic acid 940 0.400 mg EXAMPLE 7 Sodium hydroxide solution, 15% strength 0.900 mg O Glycerol 15.000 mg Investigation of the Intensification of Action of Methyl 4-hydroxybenzoate 0.150 mg insulin Purified water to 100.000 In The action of cyclopeptides of the formula I on nasal absorption of human insulin or other insulin was investi gated, after application of the test substances according to 15 Example 4, on male rats having a body weight of 100 g and EXAMPLE 11 anesthetized by ether. The decrease in the blood glucose level serves as a parameter for the action. It was found that peptides PMB-to-III in a concentration 20 of 0.01 M were able to improve nasal absorption of 20 IU/kg Suppository of human insulin by a factor of 3-4. Salmon calcitonin 0.200 mg EXAMPLE 8 Peptide PMB-III 1.000 mg Suppository base (hard fat) to 2.500 g Investigation of the Mucosa Tolerability 25 The tolerability of cyclopeptides of the formula I which are used as auxiliaries for increasing absorption can be tested on the isolated gastric mucosa of the guinea pig. EXAMPLE 12 Wirth K, Bickel M & Deutschlander N (1987): Patent Blue 30 permeation through the isolated guinea pig gastric mucosa: a quantitative method for the assessment of gastric irritants. Diagnostic Med. Sci. Res. 15, 881-882). The bovine nasal mucosa is used in a similar model. Ditzinger G, Sandow J, Merkle HP (1990): In vitro model for nasal peptide delivery: Enhance 35 Protirelin 0.050 mg ment effects of cyclopeptides on the transport rate of two Gonadoliberin 0.025 mg Sonatoliberin 0.025 mg oligopeptides. Proceed. Intern. Symp. Contr. Rel. Bioact. Peptide PMB-I 0.250 mg Mater. 17, 220-221). The permeation of Patent Blue as a Citric acid - HO 0.170 mg marker substance for mucosa damage was investigated using Disodium monohydrogen phosphate - 12H2O 1.100 mg the latter model. While bile acids used as absorption enhanc 40 Sodium chloride 0.600 mg ers, such as sodium deoxycholate or sodium glycocholate, or Benzyl alcohol 1.000 mg surfactants used as absorption enhancers, such as poly Purified water to 0.00. In docanol (Laureth-9), increased the permeation of Patent Blue by about 300%, peptides PMB-I to -III show no We claim: influence on the rate of permeation. 45 1. A pharmaceutical formulation comprising EXAMPLE 9 a) a pharmacologically active amount of one, two or three peptides in each case comprising 3 to 51 amino acids, Nasal Solution 50 or physiologically tolerated salts thereof, and Buserelin 0.15 mg b) at least one additional peptide, said additional peptide Peptide PMB-I 1.50 mg being Peptide PMB-I: Sodium chloride 0.80 mg Citric acid. HO 0.11 mg Sodium citrate 2H2O 0.15 mg 55 Benzalkonium chloride 0.01 mg Disodium EDTA 0.01 mg Water (purified) to 0.1000 In 5,565,423 12 2. At least one peptide, said peptide being a Peptide PMB-I PMB-I:

PMB-I - Dab - G LeuN Dab D-Phe

10 N ThrY/ Dab

15 N he-co-ab-UU Nis PMB-I 20 Peptide PMB-II:

PMB

25

30 Dab-CO OH,

OH, OH 35 Peptide PMB-III: OH Dab - G Leu PMB-III Peptide PMB-III: N Dab -Ge. Leu PMB-III Dab 4. D-Phe - N Thr Dab Y/ 45 / ripal R-" Dab -CO-CH2 50 ripal Dab-CO-CH2 or a physiologically tolerated salt of any of said additional peptides. or a physiologically tolerated salt thereof. 55

60

65 5,565,423 13 14 3. The compound of the formula Peptide PMB-I: or a physiologically tolerated salt thereof. 5. The compound of the formula Peptide PMB-III: Dab - G Leu / N. Dab - G Leu Dab D-Phe 5 - N Y / Dab D-Phe Thr Dab 10 Thr Dab N /co N Y-Dab N Disaros CO 2 A W *D, s^s. 15 Dab-CO-CH or a physiologically tolerated salt thereof. 4. The compound of the formula Peptide PMB-II: or a physiologically tolerated salt thereof. 20. 6. A method for promoting the absorption of peptides by Dab -Ge. Leu the mucosa, wherein said method comprises the step of applying to the mucosa an effective amount of at least one compound as claimed in one of claims 2-5, and at least one Dab - ND-Phe peptide which has 3 to 51 amino acids and is capable of 25 being absorbed by the mucosa. 7. A pharmaceutical formulation as claimed in claim 1, Y. Dab which comprises a peptide containing 3 to 51 amino acids. 8. A method for promoting the absorption of peptides in 30 application to the mucosa, wherein said method comprises: YDb applying to the mucosa an effective amount of at least one Dab-CO OH compound as claimed in one of claims 2-5 and at least one peptide which has 3 to 51 amino acids; and absorption of said peptide by the mucosa. OH 35 ck k :k k k