()Acial-On O 4,153,689 5/1979 Hirai Et Al

USOO5397771A United States Patent 19 11 Patent Number: 5,397,771 Bechgaard et al. 45 Date of Patent: Mar. 14, 1995

(54 PHARMACEUTICAL PREPARATION Administration in Solution and by Suppository,’ Acta 75 Inventors: Erik Bechgaard, Hellerup, Denmark; Paediatr Scand, 66 (1977) pp. 563-567. Sveinbjorn Gizurarson, Keflavik, DeFlines, "Bereiding van een diazepam-clysma, Phar Iceland; Rolf K. Hjortkjaer, maceutisch Weekblad, 114 (1979) p. 805. Humlebaer, Denmark Spiegelbert et al., "Ein Neues Injizierbares Lösungsmit tel (Glycofurol)” 6 Jahrgang, s.75. 73 Assignee: Bechgaard International Research Moolenaar et al., "Biopharmaceutics of Rectal Admin and Development A/S, Hellerup, istration of Drugs in Man IX... ', International Journal Denmark of Pharmaceutics, 5 (1980) pp. 127-137. 21 Appl. No.: 118,683 (List continued on next page.) 22 Filed: Sep. 10, 1993 Primary Examiner-Lester L. Lee Related U.S. Application Data Assistant Examiner-A. M. Davenport 63 Continuation of Ser. No. 791,651, Nov. 14, 1991, aban Attorney, Agent, or Firm-Wegner, Cantor, Mueller & doned, which is a continuation-in-part of Ser. No. Player 696,564, May 8, 1991, abandoned. 30 Foreign Application Priority Data May 10, 1990 DK Denmark ...... 1170/90 Aug. 30, 1990 DK Denmark ...... 2O75/90 57 ABSTRACT 51) Int. Cl...... A01N 37/18: A61K 37/00; A pharmaceutical preparation for application of an C07K 5/00; CO7K 7/00 effective amount of one or more biologically active 52 U.S.C...... 514/2; 514/4; Substance(s) to a mucosal membrane of a mammal com 514/3; 530/303; 530/307; 530/311; 530/313 prising an n-glycofurol represented by the formula I: 58 Field of Search ...... 514/2, 3, 4: 530/303, i 530/307, 311, 313 56) References Cited U.S. PATENT DOCUMENTS ()acial-on O 4,153,689 5/1979 Hirai et al. . FOREIGN PATENT DOCUMENTS wherein n is 1 to 4 in an amount from: 0.1-30% prefera 95897 12/1983 European Pat. Off.. bly 0.1-20% most preferably 1-15% in water, or in 183527 6/1986 European Pat. Off. . vegetable oil or n-ethylene glycol(s) represented by 278997 8/1988 European Pat. Off. . formula II: 418642 3/1991 European Pat. Off. . 4301.49 6/1991 European Pat. Off. . HOCH2CH2OH 2528516 2/1976 Germany . OTHER PUBLICATIONS wherein p is 2 to 8, or in a mixture thereof. Nasal admin Jeppsson et al., “Anticonvulsant Activity in Mice of istration of the preparation produces a high plasma Diazepam in am Emulsion Formulation for Intravenous concentration of the pharmaceutically active substan Administration,” Acta Pharmacol. et Toxicol, 36 (1975) ce(s) nearly as rapid as by i.v. administration. pp. 312-320. Knudsen, “Plasma-Diazepam in Infants. After Rectal 29 Claims, 16 Drawing Sheets 5,397,771 Page 2

OTHER PUBLICATIONS PCT/EP89/01186-WO 90/03796 (Schiapparelli). Kromann et al., "Diazepam in an Oil Emulsion', Letters PCT/US89/05260-WO 90/06136 (Schering). to the Editor, 61, No. 6 (1982) p. 544. Vromans et al., "Effect of Solvents on Rectal Absorp Moolenaar et al., “Rectale Irritatie van Vehiculae ... ', tion Rate of Paracetamol in Man: an in vitro Ap Pharmaceutisch Weekblad, 116 (1981) pp. 33-34. proach', International Journal of Pharmaceutics, 26 Yasaka et al., “Mechanisms in the Potentiation and (1985) pp. 5-13. Inhibition of Pharmacological Actions of Hexobarbital Moolenaar et al., "Rectal Versus Oral Absorption of and Zoxazolamine by Glycofurol,” Biochemical Phar Diflunisal in Man', International Journal of Pharmaceu macology, 27 (1978) pp. 2851-2858). tics, 19 (1984) pp. 161-167. . Illum, "Drug Delivery Systems for Nasal Application', Moolenaar et al., "Rectal Absorption of Methadone Archiv for Pharmaci og Chemi, 94 (1987) pp. 127-135. from Dissolution-Promoting Vehicles”, International Morimoto et al., "Nasal Absorption of Nifedipine from Journal of Pharmaceutics, 33 (1986) pp. 249-252. Gel Preparations in Rats', Chem. Pharm. Bull, 35/7, Lashmar et al., “Topical Application of Penetration (1987) pp. 3041-3044. Enhancers to the Skin of Nude Mice: a Histopathologi Lau et al., "Absorption of Diazepam and Lorazepam cal Study,” Journal of Pharmaceutics, 41 (1988) pp. Following Intranasal Administration', International 118-121. Journal of Pharmaceutics, 54 (1989) pp. 171-174. Moolenaazr et al., "Manipulation of Rectal Absorption PCT/US89/03897-WO 90/02737 (Goldberg). Rate of Phenytoin in Man,' Pharmaceutisch Weekblad PCT/AT87/00015-WO 87/05210 (Burghart). Scientific Edition, 3 (1981) pp. 175-180. U.S. Patent Mar. 14, 1995 Sheet 1 of 16 5,397.771

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NYNY YYYYYYY NY. NNNNNNNN NYNY YYYYQ YNYNYN NYS NYS NYNY YYY NY N N N N N N N N NNN N N N N N N N NYNY YNNYNY NYYYYYYYYQYN SNNNNNNN NS NYSYNYNYSYNY YNY Y S3 NNNNNNNNN N is N N N N N N N N N > SNNNNNNNNN SYYYYYYY Ya N to NYSYNY YYYYYYQ- to SNNNNNNN st SNNNNNNN ro SNNNN N N N (n SN N N N N N N - O) o N to to St. No C - C (Uu)00UOdsel O. eul, UD9W 5,397,771 1. 2 very often it is not possible to dissolve a clinically rele PHARMACEUTICAL PREPARATION vant amount in the relatively small volume which may be applied for intranasal administration. This application is a continuation of United States For liquid compositions it is essential that an effective application Ser. No. 07/791,651, filed Nov. 14, 1991, amount of the biologically active substance(s) can be now abandoned; which is a continuation-in-part of Ser. dissolved in a volume of less than about 300 pull. A larger No. 07/696,564, filed May 8, 1991, now abandoned. volume can be disagreeable to the patient and will evi The present invention relates to pharmaceutical com dently drain out anteriorly through the nostrils or poste positions for administration of a biologically active riorly toward the pharynx. The result is that a part of substance to a mammal via a mucosal membrane. O the active substance is lost from the absorption site and The administration by injection (intravenous, intra that in practice it will be impossible reproducibly to muscular and subcutaneous) of biologically active sub administer a correct dose. The volume for human adults stances is normally regarded as the most convenient is preferably from about 1 pull to about 1000 ul and more way of administration when the purpose is to achieve a preferably from about 50 ul to about 150 ul per nostril. rapid and strong systemic effect, e.g. within 3-5 min 15 The mucosal epithelium in the nasal cavity is covered utes, and when the active substance is not absorbed or is with many hair-like cilia being an important defense inactivated in the gastrointestinal tract or by first-pass mechanism for the mammal body against inhaled dust, hepatic metabolism. However, the administration by allergens and microorganisms. The normal half-time for injection presents a range of disadvantages. Thus it non-absorbed substances administered to the nasal cav requires the use of sterile syringes and may cause pains 20 ity is about 15 minutes due to the mucociliary clearance and irritations, particularly in the case of repeated injec removing foreign particles and excess mucus toward tions, including the risk of infection. Besides, injections the pharynx. For this reason it is preferred that the cannot be administered by untrained persons. absorption occurs rapidly and preferably within 1 to 20 Intranasal administration is currently receiving spe minutes. cial interest, attempting to avoid the inconveniences 25 A variety of vehicle systems for the nasal delivery of caused by the direct invasion into the organism in con biologically active substances has been developed. Up nection with parenteral administration. Furthermore, to date the literature has suggested that uptake of bio this route of administration may conveniently be used as logically active substances from the nasal mucosa may an alternative to parenteral injection, when a rapid be made possible by incorporation into the formulation onset of effect is needed, and it can be performed by an 30 of a special vehicle system or by the addition of certain untrained person. absorption enhancing agents. In order to be an attractive alternative to injection, Lau and Slattery (1989) studied the absorption char intranasal administration should offer a similar relation acteristics of diazepam and lorazepam following intra of dosis to plasma concentration and should not cause nasal administration for the treatment of status epilepti any considerable pain or irritation to the patient nor any 35 cus. In order to solubilize these drugs, a non-ionic sur irreversible damage or irritation to the nasal mucosa. factant. polyoxyethylated castor oil, was selected as the However, in the case of treatment of acute health least irritating out of several solvents studied including threatening indications, a relatively high local irritation polyethyleneglycol 400. Diazepam absorption was 84 to the mucosa may be acceptable. and 72%, respectively, in two adults measured over a In nasal administration, the biologically active sub period of 60 hours. However, the peak concentration stance must be applied to the mucosa in such a condition was not observed until 1.4 hours after the nasal adminis that it is able to penetrate or be absorbed through the tration and was only about 27% with reference to i.v. mucosa. In order to penetrate the mucus the vehicle administration, which indicate that most of the absorp must be biocompatible with the mucus and hence have tion had probably taken place after the test substance a certain degree of hydrophilicity. However, the vehi 45 was cleared down to pharynx and swallowed. Similar cle should preferably also posses lipophilic properties in results were obtained for lorazepam giving an even order to dissolve a physiologically active amount of longer time to peak (2.3 hours). The authors conclude certain biologically active substances. that the intranasal route of administration had limited The extensive network of blood capillaries under the potential for the acute treatment of epileptic seizures. nasal mucosa is particularly suited to provide a rapid 50 Wilton et al. (1988) attempted to administer midazo and effective systemic absorption of drugs. Moreover, lam to 45 children for achieving preanaesthetic seda the nasal epithelial membrane consists of practically a tion. The volumes used were very impractical and ex single layer of epithelial cells (pseudostratified epithe ceeded the maximal volume required for efficient ad lium) and it is therefore more suited for drug administra ministration resulting in coughing and sneezing with tion than other mucosal surfaces having squamous epi 55 expulsion of part of the dose. The paper does not de thelial layers, such as the mouth, vagina, etc. These scribe the aqueous vehicle system used. surfaces, however, are also suited for the application of International Patent Publication No. WO 86/04233 biologically active substances using the drug delivery discloses a pharmaceutical composition wherein the system according to the invention. drug (e.g. diazepam) is dissolved in a mixture of propel The effective nasal absorption is considered very lant and co-solvent e.g. glycerolphosphatide. This com small if the biologically active substances is not avail position requires a pressurized system and at least one able in water-soluble form (Proctor, 1985). This state halogenated hydrocarbon aerosol propellant. ment puts severe limitations on the use of the biologi Morimoto et al. (1987) have studied a gel preparation cally active substances, which should be water soluble for nasal application in rats of nifedipine containing the and stable in aqueous solutions. 65 gelling agent carbopol (polyacrylic acid) in polye A large number of biologically active substances, thyleneglycol 400 (PEG 400), for achieving prolonged including drugs (such as benzodiazepines), vitamins and action and high bioavailability of the drug. A mixture of vaccines, have a limited degree of water-solubility and equal amounts of carbopol and PEG 400 was preferred. 5,397,771 3. 4 It was shown that nasal application provided higher According to a preferred aspect of the invention the bioavailability of nifedipine than after peroral adminis preparation comprises tetraethylene glycol. tration, but the plasma peak concentration was not ob The pharmaceutical composition of the invention served until after 30 minutes, and it was only S10% as preferably comprises in-glycofurols wherein n is 1 or 2. compared with intravenous administration. In accordance with a preferred aspect of the inven DK patent application no. 2586/87 discloses a phar tion there is provided a pharmaceutical preparation maceutical composition comprising an antiinflamma comprising one or more n-glycofurols and one or more toric steroid, water and only 2 to 10 volume-percent n-ethylene glycols. propylene glycol, 10 to 25 volume-percent polye The pharmaceutical composition of the invention thyleneglycol 400 and 1 to 4 volume-percent Tween 20. O may comprise a biologically active substance selected U.S. Pat. No. 4,153,689 discloses a principle for ob from the group consisting of Adrenal hormones, cortico taining a stable aqueous solution of insulin intended for steroids and derivatives such as ACTH and analogies, intranasal administration. It has a pH in the range from teracosactrin, alsactide, cortisone, cortisone acetate, 92.5 to 4.7 and it contains from 0.1 to 20 weight percent hydrocortisone, hydrocortisone alcohol, hydrocorti 15 sone acetate, hydrocortisone hemisuccinate, predniso of a stabilizing agent selected from the group consisting lone, prednisolone terbutate, 9-alpha-fluoropredniso of (a) at least one nonionic surface active agent with lone, triamcinolone acetonide, dexamethasone phos hydrophile-lipophile balance value in the range of 9 to phate, flurisolide, budesonide, toxicorol pivalate, etc.; 22, (b) a stabilizing agent selected from polyethylene Amino acids. Anorectics such as benzphetamine HCl glycol's having a molecular weight in the range from chlorphentermine HCl, etc.; Antibiotics such as tetracy 200 to 7500 and (c) mixtures of stabilizing agents men cline HCl, tyrothricin, cephalosporine, aminoglyco tioned in (a) and (b). sides, streptomycin, gentamycin, leucomycin, penicillin International Patent Publication No. 90/02737 dis and derivatives, erythromycin, etc.; Anti-allergic agents, closes nasal administration of benzodiazepine hypnotics Antibodies such as monoclonal or polyclonal antibodies in a pharmaceutically acceptable nasal carrier. The 25 &against infectious diseases; Anti-cholinergic agents such carrier may be a saline solution, an alcohol, a glycol, a as atropine base, etc.; Anti-depressents such as amitripty glycol ether or a mixture thereof. There: is no indication line HCl, imipramine HCl, etc.; Anti-emitica such as that the presence of a glycol or a glycol ether should be neuroleptica, e.g. metopimazin, antihistamins e.g. thie impart special advantages to the preparation, nor that nylperazin or anti-emetica having a regulatory effect on the presence should be critical for the administration 30 the motility of the intestine such as domperidon; Anti Other preparations for intranasal or sublingual ad epileptica and anti-spasmolytica such as clonazepam, ministration are disclosed in U.S. Pat. No. 4,746,508 diazepam, nitrazepam, lorazepam etc.; Anti-histaminic disclosing the uptake of e.g. insulin by using fusidic acid agents and histaminic agents such as diphenhydramin and derivatives as absorption promoters and in Interna HCl, chloropheniramine maleate, clemastine, histamine, tional Patent Publication No.WO 87/05210 disclosing a 35 prophenpyridamine maleate, chlorprophenpyridamine sublingual sprayable pharmaceutical preparation op maleate, disodium cromoglycate, meclizine, etc.; Anti tionally comprising PEG and requiring ethanol, diglyc hypertensive agents such as clonidine HCl, etc.; Anti-in eride and/or triglyceride offatty acid and a pharmaceu flammatory agents (enzymatic) such as chymotrypsin, tically acceptable propellant gas. bromelain seratiopeptidase, etc.; Anti-inflammatory The primary object of the invention is to provide a 40 agents (non-steroidal) such as acetaminophen, aspirin, pharmaceutical composition for application of a biolog aminopyrine, phenylbutazone, mefenamic acid, ibu ically active substance via a mucosal membrane of a profen, diclofenac sodium, indomethacin, colchicine, mammal, which composition is capable of producing a probenocid, etc.; Anti-inflammatory agents (steroidal) high plasma concentration of the pharmaceutically ac such as hydrocortisone, prednisone, fluticaSone, tive substance nearly as rapid as by i.v. administration, 45 predonisolone, triancinolone, triaxncinolone acetonide, without causing unacceptable damage to the mucosal dexamethasone, betamethasone, becomethasone, be membrane. This object is fulfilled with the composition clomethasone dipropionate, etc.; Anti-neoplastic agents of the invention. such as actinomycin C, etc.; Antiseptics such as chlor The pharmaceutical preparation of the present inven hexidine HCl, hexylresorcinol, dequalinium cloride, 50 ethacridine, etc.; Anti-tumor agents; Anti-tussive expecto tion is characterized in comprising from 0.1-30%, pref rant (asthmatic agents) such as sodium cromoglycate, erably 0.11-20%, more preferably 1-15% of one or codeine phosphate, isoprotereol HCl, etc.; Anti-viral more substance(s) selected of n-glycofurols represented and anti-cancer agents such as interferons (such as Al by the formula I: pha-2 interferon for treatment of common colds), phe 55 nyl-p-guanidino benzoate, enviroxime, etc.; Beta-adren ergic blocking agents such as propranolol HCl, etc.; Blood factors such as factor VII, factor VIII etc.; Bone ()aloasa-on metabolism controlling agents agents such as vitamine O D3, active vitamin D3, etc.; Bronchoisters clenbuterol HCl, bitolterol nesylate, etc.; cardiotonics such as digi wherein n is an integer of 1 to 4, in an n-ethylene glycol talis, digoxin, etc.; Cardiovascular regulatory hormones, represented by the formula II: drugs and derivatives such as bradykin antagonists, atrial natriuretic peptide and derivatives, hydrailsazine, angi HOCH2CH2)OH otensin II antagonist, nitroglycerine, nifedipine, isosor 65 bide dinitrate, propranolol, clofilium rosylate, etc.; Che wherein p is an integer of 3-8 or in water or in a vegeta motherapeutic agents such as sulphathiazole, nitrofura ble oil or in a mixture of water and/or n-ethylene glycol zone, etc.; CNS-stimulants such as lidocaine, cocaine, and/or vegetable oil. etc.; Corticostergids such as lacicortone, hydrocortice 5,397.771 5 6 one, fluocinolone acetonide, triancinolone acetonide, histaminic agents, Anti-hypertensiva, Anti-inflamma etc.; Diagnostic drugs such as phenolsulfonphthalein, tory agents (enzymatic, non steroidal and steroidal), dey T-1824, vital dyes, potassium ferrocyanide, secre Anti-neoplastic agents, Anti-septics, Anti-tumour tin, pentagastrin, cerulein, etc.; Dopamineric agents such agents, Anti-tussive expectorants (asthmatic agents), as bromocriptine mesylate. etc.; Enzymes such as lyso Anti-viral and anti-cancer agents, Beta-adrenergic zyme chloride, dextranase, etc.; Gastrointenstinal hor blocking agents, Blood factors, Bone metabolism con mones and derivatives such as secretin, substance P, etc.; trolling agents, Bronchodilators, Cardiotonics, Cardio Hypothalamus hormones and derivatives such as LHRH vascular regulatory hormones, drugs and derivatives, and analoguss (such as naferelin, buserelin, Zollidex, Chemotherapeutic agents, CNS-stimulants, Cortico etc.), enkephalins (DADLE, metkephamid, leucine 10 steroids, Diagnostic drugs, Dopaminergic agents, En enkephalin), TRH (thyrotropin releasing hormone),. zymes, Fibrinolytics, GABA antagonists, Gastrointesti etc.; Hypothensives, Local anesthitics such as benzocain, nal hormones and derivatives, Glutamate antagonists, etc.; Migraine treatment substances such as dihydroergot Glycine antagonists, Hypothalamus hormones and de amine, ergometrine, ergotamine, pizotizin, etc.; Narcot rivatives, Hypothensives, Local anaesthetics, Migraine ics, antagonists and analgetics such as buprenorphine, na. 15 treatment substances, Narcotics, antagonists and anal loxone etc.; Pancreatic hormones and derrivatives such as getics, Pancreatic hormones and derivatives, Parasym insulin (hexameric/dimeric/ monomeric forms), gluca pathomimetics, Parasympatholytics, Pituitary gland gon, etc.; Parasympathomimetics such as nicotine, meth hormones and derivatives, Prostaglandins, Sedatives, acholine, etc.; Parasympatholytics such as scopolamine, Sex-hormones, Spasmolytics, Sympathomimetics, Thy attopine, ipratropium, etc.;. Parcinson disease substances 20 roid gland hormones and derivative, Tranquillises, Vac such as apomorphin etc.; Pituitary gland hormones and cines, Vasoconstrictors, Vasodilators, and Vitamins. derivatives such as growth hormone (e.g. human), vaso According to another aspect of the invention, the pressin and analogues (DDAVP, Lypressin), oxytocin biologically active substance(s) is(are) selected from and analoguss, etc.; Prostaglandines such as PGA and biologically active peptides which are digested in the derivatives, PGE1 and derivatives, PGE2 and deriva 25 gastrointestinal tract. tives, PGF1 and derivatives, dinoprost trometamol, etc.; According to a more preferred aspect of the inven Protease inhibitors such as aprotinin, citrate, or a 1-anti tion, the biologically active substance is selected from trypsin etc.; Sedatives such as alprazolam, bromazepam, the group consisting of coagulation factors such as Fac brotizolam, camazepam, chlordiazepeoxide, clobazam, tor VII, Factor VIII, Factor IX and derivatives and chlorazepic acid, clonazepam, clotiazepam, cloxazolam, 30 analogues thereof; agents controlling bone metabolism delorazepam, diazepam, estazolam, ethyl loflazepate, such as Vitamine D3, active Vitamine D3, calcitonin and fludiazepam, flunitrazepam, flurazepam, flutazolam, derivatives and analogues thereof; hormones secreted halazepam, haloxazolam, ketazolam, loprazolam, by hypothalamus such as LHRH or analogues, e.g. lorazepam, lormetazepam, medazepam, midazolam, nafarelin, buserelin or Zolidex, enkephalins such as nimetazepam, nitrazepam, nordiazepam, oxazepam, 35 DADLE, metkephamid or leucine enkephalin, and oxazolam, pinazepam, prazepam, temazepam, tet TRH and derivatives and analogues thereof; hormones razepam, tofisopam, triazolam, etc.; Sex-hormones such secreted by pancreas such as insulin or glucagon and as ethinyloestradiol, levonorgestrel, FSH, LH, LTH, derivatives and analogues thereof; hormones secreted estradiol-17-beta, progesterone, norethindrone, testos by the pituitary gland such as growth hormone, vaso terone, etc.; Sympathomimetics such as ephedrine, epi pressin or oxytocin and derivatives and analogues nephrine, phenylephrine, xylometazoline, tramazoline, thereof, e.g. DDAVP or lypressin; sex-hormones such dopamine, dobutamine, etc.; Thyroid gland hormones as ethinylestradiol, levonorgestrol, FSH, LH, LTH and derivatives such as calcitonins and synthetic modifi estradiol-17f8, progesterone, norethindrone or testoster cations thereof etc.; Tranquillisers such as alprazolam, one and derivatives and analoguss thereof; and tranquil bromazepam, brotizolam, camazepam, chlordiazepeox 45 izer such as alpazolam, bromazepam, brotizolam, ide, clobazam, chlorazepic acid, clonazepam, clotiaze camazepam, chlordiazepeoxide, clobazam, chlorazepic pam, cloxazolam, delorazepam, diazepam, estazolam, acid, clonazepam, clotiazepam, cloxazolam, deloraze ethyl loflazepate, fludiazepam, flunitrazepam, fluraze pam, diazepam, estazolam, ethyl loflazepate, fludiaze pan, flutazolam, halazepam, haloxazolam, ketazolam, pam, flunitrazepam, flurazepam, flutazolam, halazepam, loprazolam, lorazepam, lormetazepam, medazepam, 50 haloxazolam, ketazolam, loprazolam, lorazepam, lor midazolam, ninetazepam, nitrazepam, nordiazepam, metazepam, medazepam, midazolam, ninetazepam, oxazepam, oxazolam, pinazepam, prazepam, temaze nitrazepam, nordiazepam, oxazepan, oxazolam, pinaze pan, tetrazepam, tofisopam, triazolam, etc.; Vaccines pam, prazepam, temazepam, tetrazepam, tofisopam or such as AIDS-vaccines, influenza virus, parainfluenza triazolam and salts, derivatives and analoguss thereof. virus, measles, polio, rhinovirus type 13, respiratory 55 According to another more preferred aspect of the syncytial virus, etc.; Vasoconstrictors such as phenyleph invention, the biologically active substance selected rine HCl, tetrahydrozoline HCl, naphazoline nitrate, from the group consisting of coagulation factors such as oxymetazoline HCl, tramazoline HCl, etc.; Vasodilators Factor VII, Factor VIII, Factor IX and derivatives and such as nitroglycerin, papaverine HCl, substance P, analoguss thereof agents controlling bone metabolism VIP (vasoactive intestinal peptide) etc; Vitamines such such as calcitonin and derivatives and analogues as vitamin-B12, folic acid, or nicotinamide. thereof; hormones secreted by hypothalamus such as The composition of the invention comprises, accord LMRM or analogues, e.g. nafarelin, buserelin or Zoli ing to a preferred aspect of the invention, a biologically dex, enkephalins such as DADLE, metkephamid or active substance selected from the group consisting of leuoine enkephalin; hormones secreted by pancreas Adrenal hormones, Corticosteroids and derivatives 65 such as insulin or glucagon and derivatives and ana thereof, Amino acids, Anorectics, Antibiotics, Anti logues thereof; hormones secreted by the pituitary allergic agents, Anti-cholinergic agents, Anti-depres gland such as growth hormone, vasopressin or oxytooin sants, Anti-dots, Anti-epileptics, Anti-histaminic and and derivatives and analogues thereof, e.g. DDAVP or 5,397,771 7 8 lypressin; sex-hormones such as FSH, LH or LTM and According to a preferred aspect of the invention the derivatives and analogues thereof. composition comprises more than 50% (w/w) of poly The present invention is of particular importance ethylene glycol having an average molecular weight when administering active substances being biologically ranging from 200 to 1000. active peptides which are digested in the gastrointesti- 5 The invention also relates to a method for treatment nal tract such as insulin, glucagon, growth hormone or of a mammal with an effective amount of a biologically insulin like growth factors or derivatives or analoguss active substance wherein the dosage unit quantity of a thereof. biologically active substance is applied to a mucosal In accordance with a more preferred aspect of the membrane of the mammal to be treated in a carrier inwantion, the active substance is a pancreatic polyp O which consists of from 0.1 to 30% of n-glycofurol rep spride hormone such as insulin and glucagon or a deriv resented by the formula I: ative or an analogue thereof. In accordance with another preferred aspect of the invention, the active substance is selected among an tiepileptica, spasmolytics and tranquilizers selected 15 from the group of benzodiazepines such as clonazepam, ()aloasa-o. diazepam, flunitrazepam, triazolamn, lorazepam, ni O trazepam or mixtures thereof. As has already been pointed out the active substance wherein n is 1 to 4, in n-ethylene glycol represented by must be present in an effective amount within a total 20 the formula II: volume of not more than 1000 ul, preferably not more than 700 ul, a preferred dosage being 50-150 ul per nostril. wherein p is 3 to 8 or in water or in a vegetable oil or in The active substance may be present in an amount of 25 a mixture of water and/or n-ethylene glycol and/or from 0.0001% to 50% of the total composition, prefera vegetable oil. bly in an amount of from 0.001% to 20% e.g. in the case According to a further aspect the invention relates to of benzodiazepins. the use of a pharmaceutical composition comprising a The pharmaceutical preparation of the invention may dosacje unit quantity effective amount of a biologically furthermore comprise nitric acid and/or nitrate in a 30 active substance, from 1-30% in-glycofurols repre concentration ranging from 0.0001 to 5%. Such compo sented by the formula I: nent may act as a stabilizer for active substances such as clonazepam. The pharmaceutical preparation of the invention may, in its broadest aspect, be considered as preparation 35 comprising from 0.10-30% of one or more n-glycofurol ()aloasa-on represented by the above formula I wherein n is an O integer of 1 to 4 and a carrier, in addition to the biologi cally active substance(s). Such carrier may comprise wherein n is 1 to 4, in n-ethylene glycols represented by water and/or n-ethylene glycol and/or propylene gly the formula II: col and/or vegetable oil and/or even powdery carrier of e.g. microspheres. HOCH2CH2OH Furthermore, some pharmaceuticals preparations described herein outside of the 0.1-30% in-glycofural(s) wherein p is 3 to 8 or in water or in a vegetable oil or in range may be advantageous for special purposes, e.g. 45 a mixture of water and/or n-ethylene glycol and/or preparations comprising a very high proportions of vegetable oil for application to a mucosal membrane. glycofurols in the range of from 65-98% preferably a The mucosal membrane to which the pharmaceutical content of glycofurol in the range of 80-95%. Even a preparation of the invention is administered may be any preparation comprising pure glycofurol and the phar mucosal membrane of the mammal to which the biolog maceutically active ingredient may be advantageous for 50 ically active substance is to be given, e.g. in the nose, special purposes. The same applies for some prepara vagina, eye, mouth, genital tract, lungs, gastrointestinal tions comprising a very high proportion of PEG e.g. tract, or rectum, preferably the mucosa of the nose, pure 4EG or 3EG, even without the presence of mouth or vagina. glycofurol. The pharmaceutical compositions of the invention According to a further aspect of the invention the 55 may be administered in the form of a sublingual lozenge pharmaceutical preparation additionally comprises or troche or a buccal or nasal spray in the form of a minor proportions of one or more compound(s) selected solution, suspension or a powder, optionally together from the group consisting of surfactants, absorption with various absorption enhancers or more preferred in promoters, water absorbing polymers, microspheres, the form of slightly viscous solution which is bioadhe oils, emulsions, liposomes, substances that inhibit enzy sive or optionally in the form of a suppository or vagi matic degradation, alcohols, organic solvents, water, tory. hydrophobic agents, pH-controlling agents, preserva According to a preferred aspect of the invention, the tives and osmotic pressure controlling agents, cyclodex application is to the nasal mucosa, and the volume of the trines and propellants or mixtures thereof. pharmaceutical composition is 1-1000 ul, preferably Preferably, the water absorbing polymers are poly 65 not more than 700 ul, say 50-150 ul per nostril, a most ethylene glycols having an average molecular weight preferred exemplary dosage being 50 ul. ranging from 200 to 7500 or propylene glycol or mix Also, 0.1-20% of the glycofurol(s) is preferred in the tures thereof. carrier composition, more preferably 1-15%. 5,397,771 10 The preferred use for the carrier compositions of the invention is in pharmaceutical preparations for systemic TABLE 1-continued administration through the mucosa of the nose, mouth Composition of PEG 200 or vagina, most preferred through the mucosa of the Diethylene glycol 3.4% Triethylene glycol 21.2% nose, e.g. as a nasal spray. Tetraethylene glycol 31.2% The n-glycofurols of the formula I are considered to Pentaethylene glycol 24.4% be a pharmaceutically acceptable carrier, especially a Hexaethylene glycol 14.0% pharmaceutically acceptable carrier for nasal adminis Heptaethylene glycol 5.4% tration. According to another aspect of the invention Octaethylene glycol 0.3% n-glycofurols of the formula I are considered as an 10 100.0% enhancer facilitating the uptake of biologically active substance through a mucosal membrane of a mammal, nEG's and glycofuroles are available in various quali especially through the mucosa of the nose. ties. Especially preferred are highly purified qualities The present invention renders it possible to normalize such as 4EG from Fluka-Chennie AG (Buchs, Switzer e.g. the blood glucose level in subjects suffering from 15 land, art. no.: 86660, 1990), abbreviated 4EGf. abnormalities in their sugar metabolism such as hypo The 4EG used in the preparations of the invention is glycaemia or hyperglycaemia by giving glucagon or preferably in a very pure form which improves the insulin or an analogue or derivative thereof respec results. The 4EG is preferably highly purified being tively, in a very easy and convenient manner. >97% pure, the more preferred >98.5% pure giving Furthermore, the invention enables very convenient 20 the best results. administration of bone metabolism controlling agents The delivery system according to the invention can such as vitamin D3 or Calcitonin and derivatives and be optimized e.g. with respect to bioadhesion, sprayabil analoguss thereof; hormones secreted by hypothalamus ity or viscosity. E.g. GF, in concentration of only 5%, such as LHRH such as nefarelin, buserelin or Zolidex has a surprisingly positive effect on the sprayability of and derivatives and analoguss thereof; hormones regu 25 e.g. 4EGf by reducing viscosity. Furthermore, the addi lating the growth or mitogenic growth factors such as tion of 5% of GF is able to decrease the solidification somatropin, IGF-1, IGF-2, FGF, PDGP, TGF, EGF, temperature from about -10 to S-20 C. This is of and derivatives and analoguss thereof; protease inhibi importance where the formulation should be carried on tors such as aprotinin, citrate, or a 1-antitrypsin deriva persons or transported outdoor, ready for use. tives and analoguss thereof; and cytochines such as IL-1 30 The pharmaceutical composition of the invention and IL-2, in order to normalize the blood level thereof may be used to treat animals such as domestic animals in case of a deficiency. or pets or, preferably, human beings. Especially preferred for use in vehicle compositions A special advantage in using the above vehicle sys according to the invention in glycofurol 75 (GF) which tem is that e.g. highly lipophilic substances such as the refers to commercially available solvents of polymers of 35 benzodiazepines as well as water soluble substances e.g. the above formula I, wherein n is mainly 1 and 2. peptides and proteins such as the pancreatic hormones (Chemical Abstract Registration No. 9004 76-6). can be solubilized in clinically relevant dose for human Glycofurol 75 is a colourless liquid miscible with water, subjects in only e.g. 25-300 ul of the vehicle. In aqueous alcohols, such as methanol, ethanol, n-propanol, glyc solutions clinically relevant doses of diazepam and erol and various oils in all proportions and has a b.p. clonazepam will alternatively have to be dissolved in about 155 C. GF is reported to cause irritation when about 5000 ml and >10 ml, respectively. used in compositions for parenteral administration undi The vehicle system according to the invention may luted form as reviewed by Spiegel and Nosewothy be used in combination with various co-solvents, such (1963). It has been reported to be non-toxic and non as vegetable oil such as Miglyol (R) 840 (Dynamit Nobel irritating when diluted in water (Spiegelberg et al., 45 Chemie, Troisdorf, W-Germany) or optionally hydro 1956). genated or ethoxylated castor oil, which surprisingly The n-ethylene glycols used in accordance with the increases the possibilities for designing a controlled present invention may e.g. be triethylene glycol (3EG), release-formulation such as a diazepam formulation tetraethylene glycol (4EG), pentaethylene glycol which avoids peak plasma concentrations. (5EG), hexaethylene glycol (6EG), heptaethylene gly 50 The composition according to the invention may col (7EG), octaethylene glycol (8EG), nonaethylene comprise one or more additional pharmaceutical exipi glycol (9EG), decaethylene glycol (10EG), undeca ents, such as: surfactants and absorption promoters hav ethylene glycol (11EG), dodecaethylene glycol ing a hydrophillic-lipophilic balance from about 6 to 26 (12EG), tridecaethylene glycol (13EG) and tetradeca and ionic as well as non-ionic surfactants including ethylene glycol (17EG). The ethylene glycols may be 55 polyoxyethylene alcohol ethers, bile salts and deriva used in the form of the single compounds or a mixture of tives thereof, fusidic acid and derivatives thereof, oleic two or more n-ethylene glycols, e.g. commercial prod acid, lecithin, lysolecitins, or Tween 20 to 85; Water ucts such as polyethylene glycol 200 (PEG 200) or absorbing polymers, such as polyethyleneglycol 200 to polyethylene glycol 400 (PEG 400). 7500, polyvinylpyrrolidone, propyleneglycol, or poly 1EG to 14EG are colourless liquids miscible with acrylic acid, gelatine, cellulose and derivatives; Sub water and alcohols in all proportions. PEG 200 is a stances which inhibit enzymatic degradation, such as, commercially available mixture or ethylene glycols citrate or aprotinin; Alcohols, such as, ethanol, glyc having an average molecular weight of about 200. The erol, or benzylalcohol; Organic solvents, such as, ethy composition is shown in Table i: lacetate, or benzylalcohol; Hydrophobic agents, such as 65 vegetabile oil, e.g. soybean oil, peanut oil, coconut oil, TABLE 1. corn oil, olive oil, sunflower oil, castor oil, Miglyol (R) Composition of PEG 200 810/812/840 or mixtures thereof; pH-controlling Monoethylene glycol 0.1% agents, such as, nitric acid, phosphoric acid, or acetic 5,397,771 11 12 acid, citrate; Preservatives and osmotic pressure con trolling agents, such as glycerol, sodium chloride, Example 1 methyl paraoxybenzoate, or benzoic acid; Powder com Toxicity and Acceptability study of the vehicles positions, such as, alfa-, beta- and gamma-cyclodex according to the invention. trines, cellulose and derivatives; Microspheres, lipo- 5 In a toxicological evaluation of the vehicle the local somes and emulsions compositions, such as, starch, al as well as the systemic effect after absorption should be bumin, gelatine, or lecithins or lysolecithins; Microen considered. GF and PEG 400 is used as excipient in capsulated formulations; Propellants such as butane; injection formulations, where the administered amount Water. The use of alcohols or propellants are not man is greater than 300 ul per dose, which exceeds the datory in the composition according to the invention. 10 amount administered intranasally using the composi The invention is further illustrated with reference to tions according to the invention. the accompanying drawings in which Local toxicity related results for nEG and GF after FIG. 1 shows a graphical representation of the mean intranasal administration are not available from the plasma concentration of Clonazepam after intravenous literature. Therefore, tests were conducted assessing the administration and intranasal administration of a prepa 15 ration in accordance with the invention, local toxicity of 30-100 ul to rabbit nasal mucosa. In FIG. 2 shows a block diagram of the mean time to these tests benzodiazepines were also dissolved in vari response as a function of the contents of glycofurol in a ous solvent systems according to the invention and 30 prepartion according to the invention, ul of the composition was applied daily for 14 days to FIG.3 shows a graphical representation of the blood the nasal mucosa of rabbits. The effect of the thus ad glucose level after administration of insulin in accor ministered compositions was then assessed in five exper dance with the invention, iments including a control carried out as stated in Table FIG. 4 shows a graphical representation of the blood 2: glucose level after administration of glucagon in accor TABLE 2 dance with the invention, 25 Assessment of local toxicity of benzodiazepine compositions FIG. 5 shows a graphical representation of the con according to the invention. centration of Clonazepam in plasma after administration Animal Left Nostril of a preparation comprising various ethylene glycol No/Sex Group Test Material Session/Day constituents, 1 m 1. 30 pil Saline I FIG. 6 shows a graphical representation showing the 30 2 Control mean plasma level of diazepam after administration of 3 4f preparations comprising glycofurol and various sol 5 Vents, 6 FIG. 7 shows a graphical representation of the effect 7 In 2 30 ul D in PEG of various contents of glycofurol in preparations ac 35 8 9 cording to the invention, 10 f FIG. 8 shows a graphical representation comparing 11 the plasma level of flunitrazepam after i.v. administra 2 tion and intranasal administration according to the in 13 m 3 30 ul L in PEG 1. 40 4. vention, 15 FIG. 9 shows a graphical representation comparing 16 f the plasma level of clonazepam after i.v. administration 17 and intranasal administration according to the inven 18 19 in 4 30 ul F in PEG tion, 20 FIG. 10 shows a diagram showing the stability of 45 21 clonazepam in various vehicles, 22 f FIG. 11 shows a graphical representation comparing 23 the plasma concentrations of estrogen after i.v. adminis 24 25 m 5 30 ul F in PEG + GF 1 tration and intranasal administration according to the 26 invention, 50 27 FIG. 12 shows a graphical representation comparing 28 f the plasma concentrations of estrone after i.v. adminis 29 tration and intranasal administration of estrogen ac 30 Abbreviations: cording to the invention, D = Diazepam 3%; L = Lorazepam 5%; F = Flumitrazepam 1%; PEG = Poly FIG. 13 shows a block diagram of the influence of 55 ethylene glycol 200; GF = Glycofurol 75; PEG + GF = 70% PEG + 30% GF. glycofurol on the mean time to response after intranasal Assessment of local toxicity of vehicles of compositions application according to the invention, according to the invention. FIG. 14 shows a summary of the mean results for test " - Right Nostill 1 from Table 1-5, No/Sex Group Material Session/Day FIG. 15 shows a summary of the mean results for test 60 1 in A 100 pil Saline 3 2 Control 2 from Table 1-5, and 3 FIG. 16 shows a summary of the mean results for test 4 f 2 from Table 1-5. 5 The invention is explained more in detail with refer 6 ence to the Examples which are to be considered only 65 7 m. B 30 ul PEG 1. 8 C 100 ul PEG 1 as illustrating the invention and not to be construed so 9 D 100 pil PEG 3 as to limit the scope of the invention as set forth in the 10 f E 30 ul PEG + GF 1. appended claims. 11 F 100 ul PEG + GF 5,397,771 13 14 TABLE 2-continued was above upper normal limit, but this was considered 12 G 100 il PEG - GF 3 without clinical significance. 3 In B 30 ul PEG 1. Study procedure 14 C 100 ill PEG 1. 15 D 100 ul PEG 3 The study was a double blind test of three test vehi 16 f E 30 ul PEG + GF 1. 17 F 100 ul PEG - GF 1. cles and a control (saline) and consisted of three identi 18 G 100 ul PEG - GF 3 cal sessions, with 24 hours between each session. 19 m E 30 ul PEG + GF l During each session a control vehicle was adminis 20 F 100 pill PEG + GF tered into the right nostril 5 minutes prior to application 2 G 100 ul PEG + GF 3 10 22 f B 30 ul PEG 1. of a test vehicle into the left nostril. The subjects were 23 C 100 ul PEG 1. asked to record in a questionnaire any pain or irritation 24 D 100 ul PEG 3 that was observed 0-1 min. after application, and 10 and 25 m E 30 ul PEG -- GF 1 30 minutes after application of the test vehicle. If any 26 F 100 ul PEG + GF 27 G 100 ul PEG + GF 3 15 symptoms were noted, the subject were to record the 28 f B 30 ul PEG severity and to characterize the symptom(s). 29 C 100 ul PEG Prior to application of vehicles and 30 minutes after 30 D 100 ul PEG 3 application of the test vehicle the mucous membrane in Abbreviations: both nostrils were inspected by the experimentor by A = Control; PEG = Polyethylene glycol 200; GF = Glycofuro. 75; PEG -- GF - 70% PEG -- 30% GF. 20 nasoscopy. Vehicles The nasal cavity was examined histologically and only mild inflammatory changes were found after the admin Four vehicles were tested, one being saline and used istration of the compositions. No clear dose related as a negative control. The other three vehicles were response was found with increasing doses and no differ 25 two test vehicles according to the invention (Vehicle A ence between 100% PEG-200 and 70% PEG and B) and a "positive control” which is the vehicle of 200-30% GF was found. It was concluded that the a very well tolerated marketed product, Locilan (R) vehicles according to the invention will only incur mild (Nielsen et al., 1989). reversible toxicological effects. Devices The tolerance to two test vehicles containing tetra 30 ethylene glycol (4EGf) and optionally 5% glycofurol Pfeiffer pump (model 6917.5902/37900129) unit de (GF) was studied after intranasal application in nine livering 50 when activated was used for application. healthy volunteers, and compared, after a double blind Dosage three way cross-over design, with the vehicle of a mar keted product, Locilan (R) (Astra-Syntex Scandinavia 35 The vehicles were applied after placing the spray unit AB, Södertälje, Sweden). As control saline (0.9% so of the Pfeiffer pump in the left (or right) nostril and dium chloride) was used. activating the pump twice, the total dose being 100 pil. Saline and the vehicle of the marketed product were Clinical inspection very well tolerated. The two vehicles containing tetra ethylene glycol caused burning sensation or stinging The clinical inspection of the tissue in the nose did and runny nose of moderate severity immediately after not disclose any clinical significant effect of the ap application. The symptoms were short lasting, being plicated vehicles. In three of the subjects a mild degree over or only of mild severity 10 min. after application. of local irritation was noted 30 minutes after application No clinical symptoms could be observed by nasoscopy of the vehicles; the vehicles applied being saline in one 30 minutes after the application. 45 subject and test vehicle B in the other two subjects. When questioned all subjects stated, that the 4EGf Tolerance as reported by the subjects (Table 3-5) containing vehicles optionally comprising 5% GF were acceptable for administration of essential biologically The severity of irritation (symptoms) in the nose 0-1 active substances to be used occasionally, and that they min., 10 min. and 30 min. after application of test vehi would prefer intranasal application in one, of these 50 cles is summarized in table 3, 4, and 5, respectively. vehicles to intravenous administration. The subjects concluded that the tetraethyleneglycol The experimental details and results are given below. vehicles optionally comprising glycofurol were less well tolerated than saline of the vehicle in Locilan (R). Acceptability study of the vehicles according to the However, the symptoms caused by the 4EGf-contain invention in human subject. 55 ing vehicles were relatively short lasting and consid Subjects ered acceptable, if the biologically active substances were essential for the individual's health. Nine healthy volunteers were enrolled in the trial. All subjects were healthy without any cardiac, hepatic or TABLE 3 nephrological diseases. Tolerance 0-1 min. after application as reported in None of the subjects were suffering from any pulmo questionnaires by the subjects: No. of subjects having symptoms nary or respiratory diseases or had had a cold within the and severity thereof. Control last 2 weeks. Vital signs (blood pressure and pulse) were Severity of (right Vehicle within normal range for all subjects. Standard tests for irritation nostril) A. B C biochemistry and haematology were carried out within 65 No irritation 25 0 6 two weeks prior to the study. In all but two subjects, all Mild- 2 5 3 3 clinical chemical data were within normal range. In acceptable subject nos. 4 and 5 alanine aminotransferase (SPGT) Moderate severe 0 3 6 O 5,397,771 15 16 The mixture I is used to adjust the pH of II using a TABLE 3-continued pH-electrode for viscous liquids (Radiometer GK Tolerance 0-1 min. after application as reported in questionnaires by the subjects: No. of subjects having symptoms 2711). Then the mixture III (vehicle B) is prepared. and severity thereof. 2-10 ml of the vehicle is dispensed in a Pfeiffer multidis Control 5 penser (Pfeiffer Zerstäuber; 6917.5902/37900129). Severity of (right Vehicle irritation nostril) A B C Vehicle C: Unacceptable 0. 0 0. O A = 4EGf- vehicle Lokilan (E)-vehicle B - 5% GP in 4EGf 10 Propylene glycol, Ph.Eur. III C = Locilanl (R) vehicle Polyethylene glycol 400, Ph.Nord. 63 II 26 Control = (right nostri) saline. Tween 20, Ph.Eur. III 2.5 g Benzalconium chloride, DLS 86 70 mg Disodium-EDTA, Ph.Eur. 2nd Ed. 1983 10 mg TABLE 4 Butylated hydroxytoluene, Ph.Nord. 63 II, Add. 10 mg Citric acid, Ph.Eur. III, Suppl. 77 5 mg Tolerance 10 min. after application as reported in question- 15 Sodium citrate, 2H2O, Ph.Eur. III 7.65 mg naires by the subjects: No. of subjects having symptoms and Sorbitol DAK 63 2.86 g - severity thereof.- Purified water, Ph.Eur. 2nd Ed. 1981 ad 100 g Control Severity of (right Vehicle irritation nostril) A B C 2O Example 2 No irritation 27 5 5 7 Mild/ O 4. 4. 2 10 mg clonazepam was dissolved in 2 ml of vehicle B acceptable (example 1) using ultrasound to obtain a clonazepam Moderate severe O O O 0 Unacceptable O 0 0 0 concentration of B mg/ml. 50 pull of this preparation was As 4EGf-vehicle 25 administered into each nasal cavity of male New B is 5% GF in 4EGf Zealand White rabbits, held in a supine position during C = Locilan (8) vehicle and 1 minute after the application. An Eppendorph Control = (right nostril) saline. pipette was used for the application. Blood samples were obtained from the marginal ear vein at 0, 2, 5, 10, TABLE 5 30 15, 30 and 60 minutes and the clonazepam concentration Tolerance 30 min. after application as reported in question was determined by MPLC. naires by the subjects: No. of subjects having symptoms and -severty thereof.- FIG. I shows the mean plasma clonazepam concen Control tration obtained after the administration. The figure also Severity of (right Vehicle 35 shows the plasma concentration after i.v. injection into irritation nostril) A B C the marginal ear vein of the same dose (0.5 mg) of No irritation 27 8 7 8 clonazepam as Rivotril (R), injected over minute. The Mild- 0 1 2 1 acceptable figure shows that the plasma concentration after intra Moderate severe 0 0 O O nasal application is about the same or even higher, at Unacceptable O 0 O 0 40 about 2 minutes, than for an i.v. injection. A = 4EGf-vehicle B s 5% GF in 4EGf Example 3 C = Locilan (8) vehicle Control = (right nostril) saline. Pharmacodynamic response, enhanced by GF The pharmacodynamic response was tested in New 45 Zealand White rabbits after application of 50 pull of 2.5 mg clonazepam/ml vehicle into each nostril while they Vehicle A: were in sitting position. The vehicle consisted of 0, 10, 4EGf-vehicle I Tetraethylene glycol puriss p.a. 25 m 20, 30, 70 and 100% glycofurolum 75 (Roche, Lot no. Concentrated nitric acid p.a. 1 drop 50 706127) added to tetraethylene glycol (Merck, Art. II Tetraethylene glycol puriss p.a. 25 ml 808619). Clonazepam was dissolved by ultrasonifica Solution I (app. 30 drops) ad pH 3.8 tion. FIG. 2 shows the mean time to response (rabbit can The mixture I is used to adjust the pH of II employing stay in a lying position with its hind legs stretched to a pH-electrode for viscous liquids (Radiometer GK 55 one side). Number on the top of bars denote the amount 2711). 2-10 ml of II (vehicle A) is dispensed in a Pfeiffer of responders out of four tested. Non responders within multidispenser (Pfeiffer Zerstäuber, 10 minutes are calculated as 10 minutes in the mean 6917.5902/37900129). value. For intranasal administration of clonazapam about 10% GF in 4EGm has an optimal response, 60 whereas about 30% GF has a minimal response. The Vehicle B: same pattern is observed for 30% GF in 4EGf. 5% GF in 4EGf vehicle I Tetraethylene glycol puriss p.a. 25 ml Example 4 Concentrated nitric acid p.a. 1 drop II Tetraethylene glycol puriss p.a. 25 ml 0.33 mg zinc-free (monomeric) human insulin in 50 ul Solution I (app. 30 drops) ad pH 3.8 65 12.5 mM phosphate buffer (pH 7.4) containing 5% III Solution II 23.75 in glycofurol 75 was applied into one nostril of 5 sitting Glycofurol 75 purises 1.25 m New Zealand White rabbits, weighing about 3 kg. Blood samples of 50 pull were withdrawn from the mar 5,397,771 17 18 ginal ear vein after 0, 15, 30, 60 and 120 minutes, and glycofurol 75 (GF) in 4EGm; (3) 5% GF in 4EGm; (4) blood glucose level was determined. 10% GF in 4EGm; (5) 30% GF in 4EGm and (6) GF. FIG. 3 shows the mean blood glucose level as per Blood samples were obtained from the marginal ear centage of initial for the formulation and for 0.9% so vein at following time intervals: 0, 5, 10, 15, 30 and 60 dium chloride control. 100 ul of the same formulation, minutes and the clonazepam concentration was deter but without insulin, was administered to the nasal cavity mined by HPLC. of healthy volunteers. This formulation could not be FIG. 7 shows the mean plasma clonazepam concen distinguished from 0.9% sodium chloride with respect trations obtained after the intranasal administration. An to local irritation. initial peak plasma concentration can be controlled Example 5 10 dependent on the GF/4EGm ratio in the formulation. 4 mg human glucagon in 100 pull phosphate buffer (pH Example 9 4) containing 5% GF was applied into both nostrils (50 0.5 mg flunitrazepam in 100 ul vehicle was prepared ul into each) of 5 sitting hypoglycaemic New Zealand and applied to rabbits in a manner analogous to that White rabbits, weighing about 3 kg. The formulation 15 described in example 6. The following vehicle was used: was prepared by dissolving glucagon in a 7 mM phos Polyethylene glycol 200 (Merck Art. 807483). Blood phate buffer (pH 2.5), adjusting the pH to 4.0 with 0.1N samples were obtained from the marginal ear vein at sodium hydroxide and finally 5% glycofurolum 75 was following time intervals: 0, 5, 10, 15, 30 and 60 minutes added. The rabbits were made hypoglycaemic by sub and the flunitrazepam concentration was determined by cutaneous injection of 83 pug insulin one hour prior to 20 HPLC. the experiment. Blood samples were continuously with FIG. 8 shows the mean plasma flunitrazepam concen drawn from the marginal ear vein for blood glucose trations obtained after intranasal and intravenous determination. (Rohypnol (R), Roche) administration of 0.5 mg fluni FIG. 4 shows the mean blood glucose level as per trazepam. centage of initial after the intranasal application of glu 25 cagon. Example 10 Example 6 4 mg midazolam in 100 ul vehicle was prepared and Control of peak value, Pharmacokinetics applied to rabbits in a manner analogous to that de 10 mg clonazepam was dissolved in 2 ml of following 30 scribed in example 6. The following vehicle was used: vehicles: (1) 4EGf, (2) PEG-200 and (3) 4EGm (from Polyethylene glycol 200 (Merck, Art. 807483). Blood MERCK-Schuchardt) using ultrasound. 50 ul of this samples were obtained from the marginal ear vein at preparation was administered into each nasal cavity of following time intervals: 0, 5, 10, 15, 30 and 60 minutes sitting New Zealand White rabbits by means of an Ep and the midazolam concentration was determined by pendorph pipette. Blood samples were obtained from 35 HPLC. the marginal ear vein at following time intervals: 0, 2, 5, FIG. 9 shows the mean plasma midazolam concentra 10, 15, 30 and 60 minutes and the clonazepam concen tions obtained after intranasal and intravenous (Dor tration was determined by HPLC. micum (E), Roche) administration of 4 mg midazolam. FIG. 5 shows the mean plasma clonazepam concen tration obtained after the intranasal administration. At Example 11 about 2 minutes the plasma concentration is higher for Stability studies the 4EGf than for the PEG-200 and 4EGm formula In an attempt to optimize the stability of clonazepam tions. in the vehicles according to the invention, an acceler ated study was performed. The stability of clonazepam Example 7 45 was studied over one month period at 25 and 60° C., Control of peak value, Pharmacokinetics where recovery of about 90% or more at 60° C. was 3 mg diazepam in 100 pull vehicle was prepared and considered as very satisfying. applied to rabbits in a manner analogous to that de Following formulations were prepared, containing 5 scribed in example 6. The following vehicles were used: mg/ml clonazepam, except for formulation (1) contain (1) Glycofuroium 75 (GF); (2) Miglyol 840--GF (7+3) 50 ing 20 mg/ml; and (2) in 4EGf; (3) as (2) adjusted to pH and (3) Vegetable oil--GF (7-3). Blood samples were 3.5 with phosphoric acid; (4) as (2) adjusted to pH 3.5 obtained from the marginal ear vein at following time with citric acid; (5) as (2) added ethanol 9%; (6) as (2) intervals: 0, 5, 10, 15, 30 and 60 minutes and the diaze adjusted to pH 3.5 with nitric acid (less than 0.01%); (7) pam concentration was determined by HPLC. as (2) added water 1%; (8) propylene glycol (PG); (9) as FIG. 6 shows the mean plasma diazepam concentra 55 (8) adjusted to pH 3.8 with 0.04% acetic acid; (10) as (2) tions obtained after the intranasal administration. An added acetic acid 0.4%; (11) 4EGf--5%GF added initial peak plasma concentration can be controlled acetic acid 0.4% (pH about 6); (12) as (11) added nitric dependent on the GF/oil vehicle used. The plasma acid to pH 2. (13) as (11) added benzyl alcohol 3%; (14) concentration for the GF formulation at 5 minutes is as (10) added ethanol 16% and benzyl alcohol 3%; (15) about 55% of an intravenous injection of 3 mg diazepam 4EGf--5%GF adjusted to pH 3.5 with nitric acid (less as Stesolid (R) (Dumex A/S, Denmark). than 0.01%); (16) as (15) added sodium nitrate 0.04%; f17) as (15) added ethanol 5%; (18) 4EGf--5%GF ad Example 8 justed to pH 4.2 citric acid (0.1%); (19) as (15) added Control of peak value, the role of GF as an enhancer benzyl alcohol 2%; (20) 4EGf--5%GF added sodium 0.5 mg clonazepam in 100 ul vehicle was prepared 65 nitrate 0.04%. and applied to rabbits in a manner analogous to that After 2 and 4 weeks 100 ul samples were withdrawn described in example 6. The following vehicles were from the containers and analyzed quantitatively by used: (1) tetraethylene glycol (Merck) (4EGm); (2) 2% means of HPLC. Sample concentration (and recovery) 5,397,771 19 was calculated on the basis of the peak height relative to Each rabbit received only one test compound. One an external clonazepam standard. group served as a control receiving the same volume of As seen in table 7, only nitric acid, nitrate, ethanol isotonic sodium chloride saline. 10 minutes after appli and benzyl alcohol increases the stability. The content cation the rabbits were sacrificed and bled. The right of water in 4EGf, GF and PG was about 0.05%. Amaz 5 nasal cavity was then opened and evaluated macroscop ingly the stability in e.g. 4EGf was not decreased by ically. The evaluator was blind as to the dosing scheme. addition of 1% water. Further, the stability at 25 C. The left nasal cavity was dissected and fixed in neutral was the same for formulations stored in ampoules buffered formalin for histological evaluation. Macro (under N2) and in 10 ml Pfeiffer pump (model scopic and microscopic observations in each rabbit 1-16 6917.5902-37900129). 10 are given in table 8 and summarized in table 9 with FIG. 10 is a schematic diagram over the stability of respect to the four vehicles. Amazingly no macroscopic clonazepam in the various vehicles. or microscopic difference was seen between isotonic TABLE 7 STABILITY OF CLONAZEPAM AT 25 C. AND 60 C.IN VARIOUS VEHICLES. % RECOVERY AFTER2 AND 4WEEKS 2 WEEKS 4 WEEKS - Amp. (air) Spraybott. - Amp. (air) Spraybott. No. Forn. 25° C. 60 C. 25 C. 25 C. 60 C. 25° C. 1. 4EFg 20 mg/ml 98 91. 100 82 73 87 2. 4EGf 5 mg/ml 98 82 96 83 61 84 3. 4EGf pH 3.5 98 79 100 84 62 85 (Phosphate) 4. 4EGf pH 3.5 95 43 95 81 18 82 (Oxalic acid) 5. 4EGf 00 90 99 85 71 90 Ethanol 9% 6. 4EGf pH 3.8 98 88 97 95 87 95 (Nitric acid) 7. 4EGf 94 83 94 89 71 94 Water 1% 8. PG 96 69 99 96 56 92 9. PG pH 3.8 100 99 103 93 95 97 (Acetic acid) 10. 4EGf - 0.4% 102 84 102 99 67 102 (Acetic acid) 11. 4EGf -- 5% GF 101 80 100 98 63 99 Acetic acid 0.4% 12. =No. 11 pH 2 98 86 no data 97 81 no data (Nitric acid) 13. =No. 11 -- 103 85 102 99 76 96 Benzyl-OH 3% 14. =No. 10 -- 103 92 no data 100 80 no data Ethanol. 16% Benzyl-OH 3% 15. 4EGf - 5% GF 99 94 96 96 96 98 pH 3.8 (Nitric acid) 16. =No. 15 - 102 95 100 96 93 98 Nitrate 0.04% 7. =No. 15 - 96 98 99 98 87 98 Ethanol 5% 18. 4EGf -- 5% GF 95 85 94 93 67 98 pH 4.2 (Citric acid) 9. =No. 15 - 101 96 97 98 93 99 Benzyl-OH 2% 20. 4EGf -- 93 75 99 99 78 95 5% GF - Nitrate 0.04% Concentration: Formulation No. 1 = ca. 20 mg C/ml Formulation No. 2-10 = ca. 5 mg Cami Formulation No. 1-20 = ca. 5 mg C/ml

Example 12 saline and the 3 vehicles tested. Acute toxicological study 60 This study was conducted in order to observe acute TABLE 8 OBSERVATIONS OF RABBT NASAL CAVTES changes in the rabbit nasal cavity after a single dose of MINUTES AFTER INTRANASALAPPLICATION OF vehicle in each nasal cavity. Vehicles tested were VEHICLE glycofurol 75, tetraethylene glycol (Fluka) and propy N.A. = No Abnormalities L = Level lene glycol (DLS-quality, Mecobenzon, Denmark). 65 Macroscopic Microscopic 16 rabbits were randomized into 4 groups of 4 rabbits Observations Observations each.3 groups received a single intranasal application of (Right Cavity) (Left Cavity) one of the test vehicles, 50 ul into each nasal cavity. 1. N.A. L. 1 Congestion 5,397,771 21 22 TABLE 8-continued TABLE 8-continued OBSERVATIONS OF RABBT NASAL CAVITIES OBSERVATIONS OF RABBIT NASAL CAVITIES MINUTES AFTER INTRANASALAPPLICATION OF MINUTES AFTER INTRANASALAPPLICATION OF VEHICLE VEHICLE N.A. = No Abnormalities L = Level N.A. s No Abnormalities Le Level Macroscopic Microscopic Macroscopic Microscopic Observations Observations Observations Observations (Right Cavity) (Left Cavity) (Right Cavity) (Left Cavity) L 2 Focal lymphoid aggregates L 2 Congestion, edema, submucosal L3 N.A. 10 lymphoid aggregates L 4 N.A. L 3 Congestion, edema, submucosal 2. Slightly L 1 Congestion lymphoid aggreates hyperaemic L 2 Focal haemorrhage; congestion L. 4 Congestion and pronounced L 3 Focal lymphoid aggregates lymphoid aggregates L 4 N.A. 14. 3 mm haemorrhagic L 1 N.A. 3. N.A. Ll Congestion focus towards L 2 Congestion, edema, submucosal L 2 congestion 15 anterior lymphoid aggregates L 3 Congestion L 3 Congestion, edema, submucosal L 4 Focal submucousal haemorrhage lymphoid aggregates 4. Mucous lining L. 1 Congestion L 4 Congestion, edema, submucosal nasal cavity more L 2 Congestion lymphoid aggregates pale than normal L 3 Congestion and mucous 15. A small abrasion, L. 1 Congestion, edema, focal inspissation 20 pinpoint in the haemorrhage L 4 N.A. anterior area L 2 Congestion, focal haemorrhage, 5. 2 mm haemorrhagic L. 1 Congestion edema, submucosal lymphoid focus in anterior L 2 Congestion, focal haemorrhage aggregates and desquamation, edema, small L 3 Congestion, edema, mononuclear focus of chronic inflammation cell infiltrate near basal and focal lymphoid patches 25 section L 3 Lymphoid aggregates L 4 Congestion L 4 Area of chronic inflammation 16. 4 x 3 mm area of L i Congestion; edema 6. 6 x 3 mm area of L 1 Congestion and a focus of haemorrhage L 2 Congestion, edema, focal haemorrhage dequamation, minute haemorrh. towards anterior haemorrhage and desquamation anterior L 2 Congestion and area of L 3 Congestion, edema, submucosal submuccosal haemorrhage 30 lymphoid aggregates, especially L 3 N.A. near base L 4 Slight congestion L 4 Edena, submucosal lymphoid 7. N.A. Ll Congestion aggregates L 2 Edena, congestion, focal haemor L 3 Congestion, focal lymphoid aggr L 4 Diffuse acute and chronic 35 inflammation. Poly & mono nucl. TABLE 9 cell. Blood in lymphatics. SUMMARY OF RESULTS OF RABBIT NASAL 8. N.A. L. 1 Congestion CAVTES 10 MINUTES AFTER INTRANASAL L 2 Congestion with some APPLICATION OF 4 EGf, GF, PG AND 0.9% NaCl submucosa haemorrhage, edema (ARRANGED BY VEHICLE) L 3 Congestion with some N.A. = No Abnormalities submucosal haemorrhage, edema Animal Macroscopic Results Microscopic Results L 4 Congestion with some Vehicle No. (Right side) (Left side) submucosal haemorrhage, edema. Mucous inspissation submucosal 0.9% 3 N.A. Focal haemorrhage lymphoid aggregatres NaCl 6 6 X 3 mm haemorrhagic Focal haemorrhage 9. N.A. L 1 Congestion and edema focus, ant. section L 2 Congestion and edema 45 12 N.A. Focal haemorrhage L 3 Congestion and lateral wall 13 N.A. N.A. mono & polymorphonuclear cell Glyco- 2 N.A. Focal haemorrhage infiltrate with lymphoid aggreg furol 8 N.A. Focal haemorrhage L 4 Congestion and lateral wall (GF) 9 N.A. N.A. mono & polymorphonuclear cell 16 4 x 3 mm haemorrhagic Focal haemorrhage infiltrate with lymphoid aggreg 50 focus, ant. Section 10. N.A. L. 1 Congestion Propy- 4 N.A. N.A. L 2 Congestion with occasional lene 5 2 mm haemorrhagic Focal haemorrhage focal lymphoide aggregations glycol focus, ant section L 3 Edema and congestion (PG) 11 N.A. Focal haemorrhage L 4 Focal chronic mononuclear cell 14 2 Inn haemorrhagic N.A. infiltrate; dilatation of 55 focus. ant. section lymphatics Tetra- N.A. N.A. 1. N.A. L 1 Edena and congestion ethylene 7 N.A. Focal haemorrhage L 2 Edema, congestion, focal glycol 10 N.A. N.A. haemorrhage and submucosal (4 EGf) 15 Pinpoint abrasion, Focal haemorrhage lymphoid aggregates anterior L3 Diatation of venues L 4 N.A. 12. N.A. L 1 Congestions; venule dilatation, Surprisingly it has been found that the intranasal focal haemorrhage absorption of e.g. benzodiazepines, such as clonazepam L 2 Congestion; edema, focal and diazepam, in the vehicles according to the inven haemorrhage tion is very similar to an intravenous injection (i.v.). L 3 Congestion; submucosal lymphoid 65 aggregates From FIG. 1 it appears that the peak clonazepam L. 4 Congestion; submucosal lymphoid plasma concentration (tna) is reached within less than aggregates 2-3 minutes and that the plasma concentration after 2: 13. N.A. L 1 Congestion minutes (Cpl (2 min) is about 100% of that obtained by 5,397,771 23 24 i.v. administration. The choice of the quality of the intranasal application occupied about 5 seconds. During vehicles, according to the invention, had surprisingly an and about 1 minute after the intranasal applications, the influence on the rate of absorption. The plasma concen rabbits were held in a supine position. tration of clonazepam at 2 minute is e.g. about 40% Blood samples were collected 0, 5, 10, 20, 30, and 60 higher for 4EGf (quality from Fluka-Chemie AG) than minutes after the administration. Plasma was isolated for 4EGm (quality from MERCK-Schuchardt) or PEG and stored at +20 C. until analysis. 200 (from MERCK-Schuchardt), and tina is about The plasma concentrations of unconjugated E2 and S2, 10 and 15 minutes, respectively (FIG. 5). E1 were measured by radioimmunoassay as described The pharmacodynamic response was studied in rab by Emmet et al., 1972. After extraction with ether sepa bits. The time to response after intranasal application of 10 ration of E1 and E2 was performed on columns of Se 0.25 mg clonazepam in 100 ul of vehicle was measured. phadex LH20. Radioimmunoassay was performed after The mean times are given in FIG. 2 and the experimen evaporation of the solvents. The detection limits for E1 tal details are stated in Example 3. As shown in FIG. 2, and E2 were 40 pmol/1. The intra- and interassay vari a pharmacodynamic response after intranasal adminis ance for E1 were 7.0% and 9.6%, and for E2 7.4% and tration of clonazepam in the vehicles according to the 15 10.5%, respectively. invention may be obtained in less than 2 minutes after Dose/weight correction for individual plasma con the application. centrations was calculated relative to the mean of i.v. The compositions of the invention are stable. This has dose/weight (22.3 g/kg). For formulation 1 i.v. the e.g. been demonstrated by measuring benzodiazepine area (AUC) under the E2 plasma concentration-time concentration of the compositions according to the curves, from 0 minto infinity, a one compartement with invention after a month at 60° C. and at 25 C., respec 30 seconds infusion was fitted to the data. All other tively (see example 11). A stable solution (recovery AUC's were calculated by means of the trapezoidal about 290%) can be obtained using PG after addition of rule. acetic acid (adjusted to pH 3.8-4). The same amount of The plasma concentrations of E2 and E1 after admin acetic acid was not able to render the 4EGf solution 25 istration of E2 appear from Table 10 and 11 and FIGS. stable, and the amount of acetic acid needed for the 11 and 12, respectively. FIG. 11 shows the mean S.D. adjusting pH to about 4 was too high. Amazingly ad estrogen (E2) plasma concentrations after administra justing pH using nitric acid resulted in stable solutions tion of about 50 ug estrogen to rabbits (n=3) as an of clonazepam in e.g. 4EGf optionally comprising 5% i.v.-injection (formulation 1) or intranasal administra GF (recovery about 290%). 30 tion (2 in. and 3 in. formulated with glycofurol (GF) Surprisingly, it has also been found that using the 100% and 30%, respectively). vehicles according to the invention together with insu lin or glucagon a pronounced biological response is X form. l. iv., -O- form. 2 in., and . . . o. . . obtained within 15-30 minutes after intranasal applica form. 3 in., tion to rabbits (FIG. 3 and 4, and examples 4 and 5). 35 and FIG. 12 shows the mean -S.D. estrone (E1) Example 13 plasma concentrations after administration of about 50 Bioavailability ug estrogen to rabbits (n=3) as an i.v.-injection (formu The bioavailability of estrogen (E2) and estrone (E1), lation 1) or intranasal administration (2 in. and 3 in. after intranasal (in.) application to rabbits (n=3) of a 40 formulated with glycofurol (GF) 100% and 30%, re single dose of 50 ug E2, was studied in pilot. Two for spectively). mulations containing 30% and 100% glycofurol (GF), respectively, were tested in. relative to an i.v.-injection -x-form. 1 iv., O-form. 2 in., and . . . o. . . of the same dose. form. 3 in. Materials 45 17-3-estradiol (estrogen=E2) was obtained from As seen from FIGS. 11 and 12 and Table 12, intrana Novo Nordisk (Copenhagen, Denmark), propylen sal application was very similar to i.v. administration. glycolum ad infundibilia (PG) from Mecobenzon (Co The bioavailability appears from Table 12. The bi penhagen, Denmark) and glycofurolum 75 (GF) from oavailability for E2 was 87 and 80%, and for E2--E1 Hoffman La-Roche (Basel, Switzerland). All other rea 50 104 and 95% for formulation 2 in. and 3 in., respec gents were of reagent or analytical grade. tively. C5 min for E2 was 109% and 95%, and for Drug preparation E2--El 118% and 105%. Tina was shorter than 5 min. The formulations for i.v.-injection and for intranasal application were prepared just before the administra TABLE 10 tion. Formulation 1 for iv. administration was prepared 55 Individual estrogen (E2) plasma concentration after administration by dissolving 2.729 mg E2 in 25.0 ml PG and then add of about 50 pig estrogen to rabbits (n = 3) as an i.v.-injection (formulation 1) or intranasal administration (2 in. and 3 in. ing 25.0 ml isotonic saline. Formulation 2 for in appli formulated with glycofurol (GF) 100% and 30%, respectively). cation was prepared by dissolving 4.938 mg E2 in 10.0 n.d. = lower than 0.04 nmol/ml. ml GF. Formulation 3, also for in. application, was E2 plasma Dose/ prepared by dissolving 4.996 mg E2 in 3.00 ml GF and Formula- Rabbit concentration (nmol/ml) after min. weight then adding isotonic saline to a total volume of 10.0 ml. tion O. O 5 10 20 30 60 pg/kg Dosing and study design i. i.v. 4. n.d. 490 37.0 9.0 15.0 4.90 22.2 Rabbits (n=3) having i.v. administration received 1.0 5 0.12 65.0 45.0 29.0 18.0 7.60 21.8 ml of formulation 1 i.v. (equivalent to 50 ug E2) as an 6 0.12 38.0 30.0 25.0 21.0 11.0 22.9 65 2. in. 1 0.19 750 26.0 16.0 1.0 3.70 20.1 ear-vein infusion during 30 seconds. Rabbits (n=3) (100% 2 0.09 500 32.0 8.0 1-0 5.20 21.3 having in. administration received formulation 2 or 3 GF) 3 n.d. 44.O 24.0 4.0 8.8 3.30 19.3 in. with an Eppendorph pipette. Each rabbit received 3. i.n. 7 0.10 410 31.0 15.0 8.7 3.90 17.5 50 pull into each nostril, equivalent to 50 pug E2. The (30% 8 0.09 64.0 32.0 18.0 10.0 3.40. 19.4 5,397,771 25 26 TABLE 10-continued Example 14a Individual estrogen (E2) plasma concentration after administration of about 50 g estrogen to rabbits (n = 3) as an i.v.-injection The experiment illustrates, in pilot scale, the influence (formulation i) or intranasal administration (2 in. and 3 in. of the various vehicles on the onset of the pharmacody formulated with glycofurol (GF) 100% and 30%, respectively). namic test in rabbits. n.d. = lower than 0.04 nmol/ml. E2 plasma Dose/ Materials and Methods Formula- Rabbit concentration (nmol/ml) after min. weight Materials for the preparation of the vehicles used are tion 10. 0. 5 10 20 30 60 pg/kg Glycofurolum from Hoffman-La Roche (Lot #706127), GF) 9 0.08 31.0 17.0 8.2 5.9 1.90 18.5 O tetraethylene glycol from Merck-Schuchardt (Art. 808619), Polysorbatum 20 Ph. Eur. (Tween 20, batch #102382) and Oleum vegetabile tenue DAK 63 (batch TABLE 11 #154491) from Mecobenzon, Copenhagen. Ethanol was Individual estrogen (E1) plasma concentration after administration supplied by Novo Nordisk A/S. 2.5 mg/ml of the active of about 50 ug estrogen to rabbits (n = 3) as an i.v.-injection 15 substance, clonazepam (Roche A/S, Hvidovre, Den (formulation 1) or intranasal administration (2 in. and 3 in. formulated with glycofurol (GF) 100% and 30%, respectively). mark, order #2746A01) was dissolved in the vehicle n.d. = lower than 0.04 nmol/ml. using a ultrasonic bath. Each rabbit had 50 ul applied in E1 plasma Dose/ each nostril corresponding to 0.25 mg clonazepam per Formula- Rabbit concentration (nmol/ml) after min. weight rabbit with one exception where the rabbit had 0.5 mg tion O. 0 5 10 20 30 60 pg/kg 20 (Table 4). 1. i.v. 4. n.d. 4.30 4.30 3.10 2.70 1.60 22.2 For intravenous application 1.0 ml of Rivotril (R) 5 n.d. 4.00 5.10 460 3.30 1.50 21.8 from Hoffman-La Roche (comprising 0.5 mg/ml, after 6 n.d. 2.20 2.60 2.80 2.30 .50 22.9 dilution, batch #B0447MFD0687). 2. in. 1. n.d. 11.00 9.90 7.10 4.80 3.00 20.1 Immediately after each application the rabbits were (100% 2 n.d. 6.20 7.40. 6.00 3.40 3.00 21.3 25 GF) 3 n.d. 8.50 9.00 5.90 4.30 1.60 19.3 tested as described in the below 3 tests: 3. in. 7 n.d. 8.10 8.90 5.80 3.90 2.30 17.5 1: Hind legs to one side (30% 8 n.d. 5.30 6.10 5.00 3.90 240 19.4 2: Hind legs stretched out backwards GF) 9. n.d. 8.80 7.50 3.80 3.70 1.90 18.5 3: The rabbit is lying in a suppine position

30 Results and Discussion TABLE 12 Table 1 indicates the 10% of GF is at least as efficient Pharmacokinetic data (dose corrected relative to mean i.v. dose) as pure GF and apparently better than the remaining for estrogen (E2) and estrone (E1) after single administration of mixtures and 4EG. about 50 ug estrogen (n = 3) as an i.v.-injection (1 i.v.) or intranasal administration (2 in. and 3 in.). Table 2 indicates that 5-7.5% of GF is better than 1% AUC (nmol ml min) is the area under the plasma 35 and 2.5% of GF. Pure GP is best and corresponds to concentration - time curve from 0 minto infinity. Table 1 of the experiment. 10% Oleum vegetabile (OV) Cn(nmol/ml) is the maximal plasma concentration. in GF reduces the time to observed effect considerably. tnax(min) is the time to maximal plasma concentration. Table 3 indicates that 5% of GF is better than 10% Observed mean % Relative to and 15% and equal to pure GF in the experiments of (dose corrected) i.v.-mean at SD 40 Tables 1 and 2. Pure 4EG and 9% of Etahnol as well as Parameter 1 i.v. 2 in. 3 in. 2 in. 3 in. 5% of Tween 20 is considerably inferior than the above. AUC E2 1302 138 1038 87 - 11 80 30 El 241 471 432 195 18 179 - 12 Table 4 indicates that application of the same dose in E2 + E 1543 1609 470 104 13 95.25 a minor volume (i.e. 25ul instead of 50 ul in each nos Cmax E2 57 62 54 109 - 29 95 : 35 tril) does not enhance the effect (as compared with El 4. 10 10 150 - 22 250 - 22 45 Table 2). 3EG is possibly better than 1, 2.5, and 5% of E2 - El 61 72 64 118 28 105-33 GF in 4EG but, with respect to toxicity, 4EG will be timax E2 5 5 5 100 - 0 100 : 0 the direct choice. E1 12 8 8 67 - 38 67 - 38 Table 5 indicates that; intranasal application (of e.g. 10% GF) may act just as quickly as intravenous injec 50 tion (in 2.8 min (in.) and 4.2 min (i.v.), respectively) in COMMENTS spite of the fact that the plasma level is higher after i.v. The solubility for E2 in GF is found to be about 230 administration. The reason for this exciting result is not mg/ml. Thus, a clinical dose of 50 ug is soluble in 0.25 known. One explanation might be that clonazepam is ul. Hence, the application of a clinical dose in an ex distributed in different manner after i.v. administration, tremely small ul-volumen is rendered possible using 55 precipitates or is released slowly to the aqueous phase GF. As the solubility decreases exponentially in combi of the blood; however, this is not consistent with the nation with water, the dose volume using GF as a vehi high plasma concentration. Another explanation might cle should e.g. be about 5ul, to avoid unwanted precipi be a very rapid rate of passing to the CNS from the tation of E2 in the nasal mucus. nasal cavity. A small dose volume is desirable in order to reduce or In short it can be emphasized that GF and 4EG ap eliminate a local irritating effect. Alternatively a non parently have an optimum mixture of about 5-10%. irritating co-solvent, e.g. vegetabile oil, may be added. This is most clear from test 1 in Table 1 and FIG. 1. The In this way a desired dose volume or delivery rate may tendency is also noticed in Tables 2 and 3 correspond also be obtained. To reduce plasma peak concentration, ing to the situation that GF in addition to showing good a limited precipitation of E2 in the mucus may also be 65 performance in itself, also shows enhancing properties desirable. in mixture with 4EG. Generally speaking, test 1 appears A anhydrous GF-formulation may be useful in acute to be the most important test as it shows relaxation of hot flushing as well as in chronic dosing. muscles already after two minutes. 5,397,771 27 28 TABLE THE INFLUENCE OF GLYCOFUROLE (% GF) ON THE TIME TO RESPONSE (minutes) AFTER INTRANASAL APPLICATION OF 0.25 mg CLONAZEPAM IN TETRAETYLENGLYCOL (4EG) TO RABBITS (n = 4). NOTICE: THE TIME (in minutes) IS MEASURED FROM THE START OF EACH APPLICATION (each application takes about 5-10 seconds). FORMU- FORMU- FORMU- FORMU. FORMU- FORMU LATION LATION 2 LATON 3 LATION 4 LATION 5 LATION 6 TEST: RABBIT 4EG 10% GFN 4EG 20% GFN 4EG 30% GF IN 4EG 70% GFN 4EG GF NO. 1 2.3 2. 3.5 6.7 2.5 2.3 II m 1.2 4.8 o 1.9 1. III 3.3 19 5. 2.6 IV 1.3 5.0 4.3 40 3.5 MEAN > 6.4 .6 >5.8 >7.8 3.4 2.4 NO, 2. I 2.7 3.7 6.3 7.0 2.9 I 2.0 - 15 0.4 m- m 9.2 V 4.8 10.0 MM MEAN > 6.2 5.2 > 10.0 >9.1 >9.3 >5.9 NO. 3 - 6.7 4.2 m 3.3

V au- 6.8 o 5.8 MEAN >10.0 >7.8 > 10.0 > 10.0 > 0.0 > 6.5 - NOT REPSONDED WITHEN THE 10 MN. TEST PERIOd. TO CALCULATE THE MEAN VALUE, ONLY 10 MINUTES ARENSERTED FOR NON RESPONoERS. TEST, HND LEGS TO ONESIDE TES 2. HND LEGS STRETCHED OU BACKWARDS TEST3. THE RABBTISLYING INASP.NE POSITION

TABLE 2 THE INFLUENCE OF GLYCOFURoLE (% GF). IN TETRAETHYLENEGLYCOL (4EG) AND VEGETABLE OIL (OV) IN GF ON THE TIME TO RESPONSE (minutes) AFTER INTRANASAL APPLICATION OF 0.25 mg CLONAZEPAM TO RABBITS (n = 4). NOTICE: THE TIME (in minutes) IS MEASURED FROM THE START OF EACH APPLICATION (each application takes about 5-10 seconds). FORMU- FORMU- FORMU- FORMU- FORMU. FORMU. LATION 1 LATION 2 LATION 3 LATION 4 LATIONS LATION 6 TEST: RABBIT 19%, GF IN 4EG 2.5% GF IN 4EG 5% GF IN 4EG 7.5% GF IN 4EG GF 10% OVN GF NO. 1 w 3.1 4.0 3.2 7.0 I 9.7 o 2.5 5.9 WOM 3.5 2.7 V o o - --- 2.7 MEAN >9.9 > 10.0 > 6.4 >8.5 3.8 >7.4 NO. 2 I o 3.9 7.8 I 2.57 6.4 m III o m m 4.5 3.3 V m 2.9 m MEAN > 10.0 > 100 >8.1 >100 4.4 >7.8 NO. 3 DO www. m 8.0 m II. m 7.4 o 4.8 V am - m 5.6 MEAN > 100 > 10.0 >10.O > 10.0 >7.8 >8.7 - NOT REPSONDED WITHIN THE 10 MIN. TEST PERIOD, TO CALCULATE THE MEAN VALUE, ONLY 10 MINUTES ARE INSERTED FOR NON RESPONDERS. TEST 1. HND LEGS TO ONESOE TEST 2. HND LEGS STRETCHED OUT BACKWARDS TEST3. THE RABBTISLYING INASUPINE POSITION

TABLE 3 THE INFLUENCE OF ETHANOL (ETOH), POLYSORBATE 20 (TWEEN 20) AND GLYCOFURoL (% GF). IN TETRAETHYLENEGLYCOL (4EG) ON THE TIME TO RESPONSE (minutes) AFTER INTRANASALAPPLICATION OF 0.25 mg CLONAZEPAM TO RABBITS (n = 4). NOTICE: THE TIME (in minutes) IS MEASURED FROM THE START OF EACH APPLICATION (each application takes about 5-10 seconds). FORMU- FORMU. FORMULATION 3 FORMU- FORMU- FORMU LATION 1 LATION 2 5%. TWEEN LATION 4 LATION 5 LATION 6 TEST RABBT 4EG 9% ETOH IN 4EG 20 IN 4EG 5% GFN 4EG 10% GFN 4EG 15% GFN 4EG NO. 1 o o 1.8 2.9 3.7 I 1.1 7. 15 1.5 III m 2.2 pre- 5.7 8.6 IV 3.2 9.6 2. 3.3 9.5 MEAN >7.8 > 6.4 >9.2 2.8 4. >8.3 NO. 2 MM 2. o 3.7 2.1 I -- 3.3 40 IV o o 3.2 MEAN > 0.0 >8.3 > 10.0 3.0 >8.0 > 10.0 5,397,771 29 30 TABLE 3-continued THE INFLUENCE OF ETHANOL (ETOH), POLYSORBATE 20 (TWEEN 20) AND GLYCOFURoL (% GF). IN TETRAETHYLENEGLYCOL (4EG) ON THE TIME TO RESPONSE (minutes) AFTER INTRANASAL APPLICATION OF 0.25 mg CLONAZEPAM TO RABBITS (n = 4). NOTICE: THE TIME (in minutes) IS MEASURED FROM THE START OF EACH APPLICATION (each application takes about 5-10 seconds). FORMU- FORMU- FORMULATION 3 FORMU- FORMU- FORMU LATION 1 LATION 2 5%. TWEEN LATION 4 LATION 5 LATION 6 TEST: RABBIT 4EG 9% ETOHN 4EG 20 IN 4EG 5%. GFIN 4EG 10% GF IN 4EG 15% GFIN 4EG

NO. 3 I --- - 3.6 -- -

II m -M ------

III m -- 8.0 - -

V ------3.8 - - MEAN > 10.0 > 10.0 > 10.0 >6.4 >10.0 > 10.0 --NOT REPSONDED WITHIN THE 10 MIN. TEST PERIOD. TO CALCULATE THE MEAN VALUE, ONLY 10 MINUTES ARE INSERTED FOR NON RESPONDERS. TEST 1. HIND LEGS TO ONESIDE TEST 2. HIND LEGSSTRETCHED OUT BACKWARDS TEST3. THE RABBTISLYING IN A SUPINE POSITION

TABLE 4 THE INFLUENCE OF FORMULATION ON THE TIME TO RESPONSE (minutes) AFTER INTRANASALAPPLICATION OF 0.25 MG CLONAZEPAM TO RABBITS (n = 4). CONTRARY TO TABLE 1, 2 AND 3, THE APPLIED VOLUME WAS 25ul TO EACH NOSTRIL INSTEAD OF 50 LL. ABBREVIATIONS: GLYCOFUR.OL (GF), TETRAETHYLENEGLYCOL (4EG) AND TRIETHYLENEGLYCOL (3EG). FORMULATION 1-3 ARE TESTED SAME DAY. NOTICE: THE TIME (in minutes) IS MEASURED FROM THE START OF EACH APPLICATION (each application takes about 5–10 seconds). FORMULATION 1 FORMULATION 2 FORMULATION 3 FORMULATION 4 TEST: RABBT 12 GFIN 4EG 2.5% GF IN 4EG 5%. GFIN 4EG 3EG NO. 1 8.4 5.4 - 1.4 II --- 3.5 8.6 1.5

III - - - - V -- -- - 3.5 MEAN >9.6 >7.2 >9.7 > 4.1 NO. 2 I - --- m 7.9 II - 3.7 - 3.9

III - - m --

V -- - m m MEAN > 10.0 >8.5 > 10.0 >8.0 NO. 3 - - m --

II - --- m

III - -- m mor

V - r -- m MEAN >10.0 > 10.0 > 10.0 > 10.0 - NOT REPSONDED WITHIN THE 10 MN. TES PERIOd. TO CALCULATE THE MEAN VALUE, ONLY 10 MIN UTES ARE INSERTED FOR NON-RESPONDERS. EST 1. HIND LEGS TO ONESIDE TEST 2. HIND LEGS STRECHED OUT BACKWARDS TEST3. THE RABBTISLYING IN A SUPINE POSITION

TABLE 5-continued TABLE 5 Time to response (minutes) after the application of 0.5 - - mg clonazepam intranasally and intravenously to rabbits Tine to response (minutes) after the application of 0.5 50 (n = 4). Abbreviations: Glycofurol (GF) and mg clonazepam intranasally and intravenously to rabbits tetraethylene-glycol (4EG). NOTICE: The time (in (n = 4). Abbreviations: Glycofurol (GF) and minutes) is measured from the start of each tetraethylene-glycol (4EG). NOTICE: The time (in application. (Each application takes about 5-10 seconds minutes) is measured from the start of each and 30 seconds for in. and iv. respectively). application. (Each application takes about 5-10 seconds intranasal Intravenous and 30 seconds for in. and iv. respectively). 55 Test: Rabbit 10% GF in 4EG Rivotril (R) Intranasal Intravenous Test: Rabbit 10% GF in 4EG Rivotril (R) Mean >7.9 >8.4 No. 1 I 1.6 1.2 - Not responded within the i0 min. test period. To calculate the mean value, only O. - 10 minutes are inserted for non-responders. I 5.6 9.0 TEST 1. HND LEGS TO ONESIDE III 1.9 3.0 TEST 2. HIND LEGS STRETCHED OUT BACKWARDS IV 2.0 3.5 60 TEST 3. THE RABBIT IS LYING IN A SUPINE POSITION Mean 2.8 4.2 No. 2 i 3.0 FIG. 13 shows the influence of glycofurol (% GF) on III 6.0 - the mean time to response (minutes) after intranasal IV - - application of 0.25 mg. clonazepam in tetraethyleneg Mean > 6.2 >8.3 65 lycol (4EG) to rabbits (n=4) (individual data, see Table No. 3 2.5 3.4 1) II 9.8 - o III 9.2 - In FIG. 13, the numbers on top of the bars denote the IV - -- amount of non-responders within 10 minutes test per 5,397,771 31 32 iod. Such bars are higher in reality, as only 10 minutes -continued are used for non-responders to calculate the mean. FIG. 14 shows a summary of the mean results for test Vehicle no.: Dose (mg): Formulation: 15 0.25 7.5% GF in 4EG 1 (hind legs to one side) from Table 1-5. The numbers 16 0.25 5%. Tween in 4EG on top of the bars denote the amount of non-responders, 5 17 0.25 1% GF in 4EG) within the 10 min. test period, relative to the amount 18 0.25 5% GF in 4EG) tested (such bars are higher in reality, as only 10 min. 19 0.25 1% GF in 4EG are used for non-responders to calculate the mean). 20 0.25 2.5% GF in 4EG The drug was applied in 50 pil instead of 100 pil as usual. 10 Vehicle no.: Dose (ng): Formulation: FIG.16 shows a summary of the mean results for test 1. 0.50 Rivotri (E) IV-injection 2 (the rabbit is laying in a supine position) from Table 2 0.50 10% GF in 4EG 1-5. The numbers on top of the bars denote the amount 43 0.25 Pure10% GF in 4EG of non-responders, within- - - - the 10 min. test period,w rela 5 0.25 70% GF in 4EG 15 tive to the amount tested (such bars are higher in reality, 6 0.25 Pure 3EG as only 10 min. are used for non-responders to calculate 7 0.25 5% GF in 4EG 8 0.25 20% GF in 4EG the mean). 9 0.25 9%, EtOH in 4EG I 8. E. 4EE.in 4EG?) 20 Vehicle no.: Dose (mg): Formulation: 12 0.25 10% VO in GF 1. 0.50 Rivotril (R) IV-injection 13 0.25 30% GF in 4EG 2 0.50 10% GF in 4EG 14 0.25 15% GF in 4EG 3 0.25 10% GF in 4EG 15 0.25 7.5% GF in 4EG 4 0.25 Pure GF 16 0.25 5%. Tween in 4EG 5 0.25 70% GF in 4EG 17 0.25 1% GF in 4EG?) 25 6 0.25 Pure 3EG 18 0.25 5% GF in 4EG) 7 0.25 5% GF in 4EG 19 0.25 1% GF in 4EG 8 0.25 20% GF in 4EG 20 0.25 2.5% GF in 4EG 9 0.25 9% EtOH in 4EG The drug was applied in 50 pil instead of 100 ul as usual. 1011 0.25 Pure2.5% 4EG.GF in 4EG?) 12 0.25 10% VO in GF FIG. 15 shows a summary of the mean results for test 30 13 0.25 6. GF in 4EG 2 (hind legs stretched out backwards) from Table 1-5. 14 0.25 15% GF in 4EG The numbers on top of the bars denote the amount of 8: 7.5% GF Ea non-responders, within the 10 min. test period, relative 17 0.25 SE35 to the amount tested (such bars are higher in reality, as 18 0.25 5% GF in 4EG?) only 10 min. are used for nonresponders to calculate the 35 19 0.25 1% GF in 4EG mean). 20 0.25 2.5% GF in 4EG The drug was applied in 50 pil instead of 100 ul as usual. Vehicle no.: Dose (mg): Formulation: 0.50 Rivotril (R) IV-injection 40 Example 15a : 8: E. SE in ES Furthermore, the additional compositions stated 4 0.25 FF below were prepared by dissolving the pharmacologi 5 0.25 70% GF in 4EG cal active ingredient in the vehicle during sonification 6 0.25 Pure 3EG and/or gentle heating. Compositions comprising water 7 0.25 5%GF in 4EG 45 are rendered isotonic using sodium chloride and ad 98 0.25 9%20% EtOH GF in in 4EG 4EG justed to a pH inr the physiological range of 4.0–7.5 10 0.25 Pure 4EG using phosphate buffer.

Active Unit Dose Unit Dose Ingredient in mg in pil GF2% 4EG % PEG H2O pH Example Salbutanol 0.5 50 10 90 Oxytosin SIE 50 5 95 Terbutalin 0.5 50 1 99 Sulphate Morphine 5 100 15 85 hydrochlo Beclonet. 0.75 40 5 95 diproprion Morphine 4. 200 20 80 Flumitrazepam 2 150 5 95 Aspirin 00 300 20 80

65 11 0.25 2.5% GF in 4EG) Example 16a 1312 0.25 10% VO intoin GF In additiono the below compositions were prepared in 14 0.25 15% GF in 4EG an analogous manner as described above. 5,397,771 33 34

Active Unit dose Unit dose Ingredient in mg in ul GF 7% 4EG % PEG H2O pH Example Aspirin 300 300 100 Terfenadin 10 300 100 Becomethas. 0.75 100 30 70% vegtab. oi dipropionat Flumitracepam 0.25 40 30 70% vegtab. oi

List of references Lau, S. W. J. and Slattery, J. T. (1989), “Absorption of diazepam and lorazepam following intranasal admin wherein p is 3 to 8, and mixtures thereof, so that upon istration.” International Journal of Pharmaceutics, administration of the pharmaceutical composition to the 54, 171-174. 15 nasal mucosal membrane, absorption of the biologically Mayer, W., Erbe, S., Wolf, G. and Voigt, R. (1974), active substance through the mucosal membrane and "Beiträge zur analytik und stabilität einiger phar into the blood stream of the mammal rapidly takes place mazeutisch interessanter 1.4-benzodiazepine.' Phar and thereby allows the biologically active substance to mazie 29, H.10-11, 700-707. exert its therapeutic effect. Morimoto, K., Tahara, H. and Morisaka, K. (1987), 20 2. The method according to claim 1, wherein n is 1 or "Nasal absorption of nifedipine from gel preparations 2. in rats.” Chemical and Pharmaceutical Bulletins, 35, No. 7, 3041-3044. 3. The method of claim 1, wherein the pharmaceuti Nielsen, N. H., Frglund, L., Bindslev-Jensen, C. and cal composition is in the form of a solution or suspen Svendsen, U.G. (1989), “A new formulation of sion or of microspheres which is administered to the flunisolide for intranasal application reduces side 25 nasal mucosal membrane in a dosage unit quantity vol effects.” Allergy, 44, 233-234. ume of 50-150 ul per nostril. Proctor, D. F. (1985), "Nasal physiology in intranasal 4. The method of claim 1, wherein said vehicle con drug administrations' in Chien Y. W. (ed.) Transna tains 1-15% of n-glycofurol. sal Systemic Medications, Fundamentals, Develop 5. The method of claim 1, wherein the vehicle com mental Concepts and Biomedical Assessments. ELS 30 prises n-ethylene glycol represented by the formula II, VIER Science Publishers, Amsterdam 1985, p. wherein p is 3 or 4. 101-105. 6. The method according to claim 5, wherein said Spiegel, A.J. and Noseworthy, M. M. (1983), "Use of n-ethylene glycol is tetraethylene glycol having a purity nonaqueous solvents in patenfetal products'. Journal of at least 97%. of Pharmaceutical Sciences, October 1963, vol. 52, 35 7. The method according to claim 1, wherein the No. 10, p. 907-927. vehicle further comprises at least one member selected Spiegelberg, H., Schläpfer, R., Zbinden, G. and Studer, from the group consisting of nitric acid and nitrates, in A. (1956), "Ein neues injizierbares losungsmittel a concentration ranging from 0.0001% to 5%. (glycofurol).” Arzneittelforschung 6, 75-77. 8. The method according to claim 1, wherein the Wilton, N. C. T., Leigh, J., Rosen, D. R. and Pandit, U. vehicle is anhydrous. A. (1988), "Preanaesthetic sedation of preschool chil 9. The method according to claim 1, wherein the dren using intranasal midazolam'. Anesthesiology, biologically active substance comprises a biologically 69,972-975. active peptide which is digested in the gastrointestinal We claim: tract. 1. A method for administering a therapeutically effec 45 10. A method according to claim 9, wherein the bio tive amount of a biologically active substance to the logically active substance is selected from the group circulatory system of a mammal comprising administer consisting of coagulation factors, agents controlling ing a pharmaceutical composition having a total volume bone metabolism, hormones secreted by hypothalamus, of 1-1000 pil to a nasal mucosal membrane of the mam hormones secreted by pancreas and derivatives and mal, the pharmaceutical composition comprising the 50 analogs thereof, and hormones secreted by the pituitary therapeutically effective amount of the biologically gland and derivatives and analogs thereof. active substance dissolved or suspended in a volume of 11. The method according to claim 1 comprising a 1-1000 pull of a n-glycofurol-containing vehicle compris biologically active substance selected from the group ing 0.1-30% of at least one n-glycofurol represented by consisting of coagulation Factor VII, Factor VIII, Fac 55 tor IX and derivatives thereof; calcitonin and deriva the formula: tives and analogs thereof; hypothalamus hormones se creted by LHRH or the analogs thereof; nafarelin, buserelin, Zollidex, DADLE, metkephamid, leucine enkephalin; the pancreatic hormones insulin and gluca ()aloasa-on gon and derivatives and analogs thereof; the pituitary O gland, growth hormone, vasopressin, oxytocin and de rivatives and analogs thereof, DDAVP and lypressin; wherein n is from 1 to 4, the vehicle further comprising the sex-hormones FSH, LH or LTH and derivatives a component selected from the group consisting of and analogs thereof. water, vegetable oil, n-ethylene glycol(s) represented 65 12. The method according to claim 1, wherein the by the formula II: biologically active substance is a benzodiazepine. 13. The method according to claim 12, wherein the biologically active substance is selected from the group 5,397,771 35 36 consisting of clonazepam, diazepam, flunitrazepam, phamid, leucine enkephalin; the pancreatic hormones triazolam, lorazepam, nitrazepam and mixtures thereof. inulin and glucagon and derivatives and analogs 14. A method according to claim 12, wherein the thereof; the pituitary gland, growth hormone, vasopres benzodiazepine is present in the pharmaceutical compo sin, oxytocin and derivatives and analogs thereof, sition in a concentration of from 0.001% to 20%. 5 DDAVP and lypressin; the sex-hormones FSH, LH or 15. A pharmaceutical composition for administering a LTH and derivatives and analogs thereof. biologically active substance through a nasal mucosal 22. A pharmaceutical composition according to claim membrane of a mammal, comprising an effective 15, wherein the biologically active substance is a ben amount of the biologically active substance(s) sus zodiazepine. pended or dissolved in a volume of 1-1000 ul of a n O 23. A pharmaceutical composition according to claim glycofurol-containing vehicle comprising 65-98% of at 22, wherein the biologically active substance is selected least one n-glycofurol represented by the formula: from the , group consisting of clonazepam, diazepam, flunitrazepam, triazolam, iorazepam, bitrazepam and 15 mixtures thereof. 24. A pharmaceutical composition according to claim ()aloasa-on 15 further comprising at least one member selected from O the group consisting of nitric acid and nitrates, in a concentration ranging from 0.0001% to 5%. wherein n is from 1 o 4, the vehicle further comprising 20 25. A pharmaceutical composition for administering a a component selected from the group consisting of biologically active substance through a nasal mucosal water, vegetable oil, n-ethylene glycols) represented membrane of a mammal, comprising an effective by the formula II: amount of the biologically active substance(s) sus pended or dissolved in a volume of 1-1000 ul of a n HOCH2CH2OH 25 glycofurol-containing vehicle consisting essentially of wherein p is 3 or 4, and mixtures thereof. 100% of at least one n-glycofurol represented by the 16. A pharmaceutical composition according to claim formula: 15 in a dosage unit volume of 1-1000 ul. 17. A pharmaceutical composition according to claim 15 which comprises 80-95% n-glycofurol. 30 18. A pharmaceutical composition according to claim 15 wherein said n-ethylene glycol is tetraethylene gly ()aloasa-o. col having a purity of at least 97%. O 19. A pharmaceutical composition according to claim 18 wherein said n-ethylene glycol is tetraethylene gly 35 wherein n is from 1 to 4. col having a purity of at least 98.5%. 26. The method of claim 2, wherein said vehicle con 20. A pharmaceutical composition according to claim tains 1-15% of n-glycofurol. 15 wherein the biologically active substance comprises 27. The method of claim 2, wherein the vehicle com a biologically active peptide that is digested in the gas prises n-ethylene glycol represented by the formula II, trointestinal tract. wherein p is 3 or 4. 21. A pharmaceutical composition according to claim 28. The method of claim 3, wherein the vehicle com 20 comprising a biologically active substance selected prises n-ethylene glycol represented by the formula II, from the group consisting of coagulation Factor VII, wherein p is 3 or 4. Factor VIII, Factor IX and derivatives and analogs 29. The method of claim 4, wherein the vehicle con thereof; calcitonin and derivatives and analogs thereof; 45 prises n-ethylene glycol represented by the formula II, hypothalamus hormones secreted by LHRH or the wherein p is 3 or 4. analogs, nafarelin, buserelin, Zollidex, DADLE, metke e