Biologically active mixtures

(54) (56) (57)

Applicant Fisons Ltd, London GB

Inventor 1) H. Cairns, 2) P M. Greenhalgh, 3) B. Howlett, 1,2) Loughborough, 3) Leicester

Representative Delmar J-Å

Description Synergistic and stable pharmaceutical composition comprising sodium chromoglycate in combination with a beta 2-selective bronchodilator

Published publications: SE 411 842 (A61K 9/72) FR 2 243 677, FR 2 397 833, GB 1 242 211, US 3 671 625

Other publications: Merck Index, Ninth Edition pp. 201, 3904, 5069, 5084, 8879. The Pharmacological Bases of Therapeutics, Sixth Ed., 1980, pp.166-68 Pharmacotherapy, Clinical Pharmacology, page 14, page 123. Medical Journal of Australia, 1974, pages 456 and 723. Rote List 1979, No 27,019 Abstract:

I. Salts of 1,3-bis- (2-carboxychromon-5-yloxy) -propan-2-ol with some long-acting or B2-selective bronchodilators.

II. Pharmaceutical mixtures or compositions comprising

a) one or more of the components 1,3-bis- (2-carboxychromon-5-yloxy) -propan-2-ol or a pharmaceutically acceptable salt thereof in combination with

b) one or more 82-selective or long-acting bronchodilators or a pharmaceutically acceptable salt thereof, and optionally

c) adjuvant, diluent or carrier.

6WiW BQ

The numbers in brackets indicate international identification code, INID code, Letters in clamps indicate international document code

452 249

The present invention relates to novel, synergistic and stable pharmaceutical compositions and to compositions containing the same.

A number of different bronchodilators (bronchodilators) have been known for many years in the relief of allergic lung disorders. In many cases, these bronchodilators have been administered systemically, ie as elixirs, tablets, capsules, injection preparations, etc. In all these dosage forms, there is a risk of prone to overdose. In addition, most bronchodilators have undesirable side effects, for example on the cardiovascular system. It may also be mentioned that the bronchodilators are usually used for the treatment of an asthma attack when this has already started; in other words, they were used non-prophylactically.

Sodium chromoglycate has been recommended for use in the treatment of allergic and other morbid conditions, especially in the treatment of asthma and rhinitis. Unlike the bronchodilators, sodium chromoglycate has no relaxation effect on the smooth muscle; but sodium chromoglycate has been used prophylactically, thus to prevent the initiation of an attack of these diseases. Presumably, sodium chromoglycate is usually not recommended as a curative agent, ie as a remedy for the symptoms during an ongoing allergy attack. In addition, unlike the bronchodilators, the sodium chromoglycate has only been administered by inhalation and for the treatment of asthma.

Sodium chromoglycate has previously been marketed as a dry powder formulation. When inhaled, this powder has, to some extent, caused bronchoconstriction to some extent. To avoid this slight bronchoconstriction, a preparation of sodium chromoglycate plus a subtherapeutic dose of the fast but short-acting, non-B2 selective bronchodilator isoprenaline sulfate has been marketed for use in particularly bronchosensitive patients. The proposal of the present invention to incorporate a full therapeutic dose of a long-acting or 62-selective bronchodilator cannot in any way be said to be based on the known incorporation of a small non-therapeutic dose of isoprenaline sulfate into sodium chromoglycate preparation.

• 452 249

It is a well-known phenomenon that patients generally do not like to take medications and thus take them irregularly and that, in case they have two separate medications, they tend to either not take one medicine at all or also incorrectly take more of it one medicine than the other.

It has now been found that mixtures of sodium chromoglycate with a specific group of B2 selective bronchodilators in the right therapeutic amount ratio have the advantage that they are both relieving (palliative) and prophylactic, are more effective, provide less side effects, can be used in lower doses. , can be administered directly to the area affected by the allergy, for example by inhalation, are active for a longer period, are more stable, are synergistic, be more easily and more willingly taken by the patient correctly, or exhibit other desirable properties when compared to a) certain bronchodilators when used alone; b) sodium chromoglycate when used alone; or c) certain other mixtures when tested in relevant pharmacological models.

Based on the mode of action of the sodium chromoglycate and the mode of action of the bronchodilators, one would expect subadditive effects. Namely, the sodium chromoglycate prevents the release of the anaphylaxis mediators; the bronchodilators could thus have a useful effect only insofar as the sodium chromoglycate is inactive.

Thanks to the compositions of the present invention, it is possible for the patient to use a single medication to obtain both prophylactic and symptomatic treatment. In addition, both drugs can be administered with less excipient amounts (usually about half the amount) than are required for the administration of both drugs separately. Thus, reducing the amount of excipient is extremely valuable, as some excipients, such as aerosol propellants, can be detrimental to patients' health, especially in the case of atopy patients.

According to the present invention, there is provided a synergistic and stable pharmaceutical composition comprising as active ingredient A, the disodium salt of 1,3-bis (2-carboxycro452249 mon-5-yloxy) propan-2-ol in combination with, such as active ingredient B,

2- (tert-butylamino) -1- (4-hydroxy-3-hydroxymethylphenyl) ethanol;

1- (3,5-dihydroxy phenyl) -2- (tert-butylamino) ethanol;

- (3,5-dihydroxyphenyl) -1-hydroxy-2 - [(4-hydroxyphenyl) -isopropylamino Jetane; or

D, L-7- {3 - [(8,3,5-trihydroxifenyletyl) amino] propyl} -1,3-dimethylxanthine;

or a pharmaceutically acceptable salt of any of these compounds, in solid form.

By the term g2-selective bronchodilator is meant a β-adrenoceptor agonist with greater affinity for β2-adrenoceptors whose stimulation results in relaxation of bronchial muscles (including arterial and uterine muscles) than to β1-adrenoceptors, whose agonists give rise to cardiac stimulation (and liposuction). To determine the B2 selectivity, the activity of the compound a) is compared as a relaxant in preparations of isolated guinea pig trachea with spontaneous tone (EC + g = the molar concentration, which gives 50% of the maximum relaxation response to isoprenaline) and b) its effect of increasing the atrial stroke rate of isolated, spontaneously striking guinea pig heart atria (ECj-θ = the molar concentration, which provides 50% of the maximum increase in stroke rate; which is produced by isoprenaline). One. The B2-selective compound usually has lower EC ^g on trachea than on atria. One can determine a selectivity ratio, which is the antilogarithm of the difference between the negative log EC ECg (trachea) and the negative log ECggg (atria). The greater this selectivity ratio, the more selective the compound for β2 adrenoceptors.

By long-acting it is meant that the duration of the compound's relaxing action on the smooth bronchial muscle in pharmacological tests is longer than the duration of the action of the isoprenaline or orciprenaline. To assess the duration, the efficacy of equally active iv doses of the compounds as inhibitors of acetylcholine- or histamine-induced bronchospasm in guinea pigs during anesthesia is compared. The duration of action of isoprenaline is relatively short in this test (the control response is again normal within 10-25 minutes). When

452 249 one has a compound of greater duration in terms of its action, it takes several times longer until the control response is normal again. The bronchodilator should preferably be both B2-selective and long-acting.

According to the present invention there is also provided a pharmaceutical composition comprising a mixture as above in the form of a pressure aerosol composition containing 1-20% by weight of active ingredients and 0.05-1.5% by weight of a surfactant.

Solid compositions are advantageous in comparison with solutions or other liquid compositions. After all, these are administered by means of sputtering devices when they are to be delivered to the patient, which is a time-consuming procedure, and for the administration of these liquid compositions, large, space-consuming devices must be employed, which must usually be connected to an external source of energy, for example electricity.

The amount ratios of the active ingredients A and B in the composition vary depending on the particular active ingredients used and depending on the particular use purpose of the composition in each case.

A suitable dose of active ingredient A for inhalation is in the range of 1-100 mg, preferably 1-20 mg (measured as sodium chromoglycate).

Appropriate dosages for the active ingredient B in and for inhalation are given below, as well as the appropriate number of parts by weight of active ingredient A (measured as sodium chromoglycate) per part by weight of active ingredient B.

452 249

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452 249

It is highly preferred that the dose of the bronchodilator is such that it provides a lasting effect rather than a rapid transient effect.

The mixture can be administered in the form of doses divided for administrations 1-6, preferably 2-4 times daily. Each dose may comprise one or more dosage units.

The specific ratios of the particular active ingredients A and B in compositions according to the invention may vary widely. However, a composition containing 0.4-400 parts by weight, preferably 2-200 parts by weight, active ingredient A (measured as sodium chromoglycate) per part by weight of active ingredient B. is preferred. Particularly preferred is a composition containing sodium chromoglycate and 1- (3,5-dihydroxyphenyl) -2- (t-butylamino) ethanol, or a pharmaceutically acceptable salt thereof, for example the sulfate.

The compositions according to the invention can be prepared by mixing the various active ingredients using conventional methods known per se.

The compositions and compositions of the present invention are valuable because they show pharmacological activity in animals; they are valuable in particular because they inhibit the release and / or action of pharmacological mediators obtained by in vivo combination of certain types of antibody and specific antigen, for example, by combination of reagentic antibody with specific antigen (see Example 27 of British Patent 1,292,601). The mixtures and compositions have also been found to inhibit degranulation of mast cells and interfere with reflex reactions in laboratory animals and humans, especially reflexes associated with lung function. Regarding the action of humans, it should be mentioned that, by a pre-administered administration of the new mixtures and compositions, inhibition of both subjective and objective changes occurs, which occurs in sensitized individuals when inhaled specifically antigen. Thus, the novel mixtures and compositions are valuable in treating reversible obstruction of the airways and / or in preventing excessive mucus secretion. They are thus valuable for

452 249 treatment of allergic asthma, so-called endogenous asthma (ie in cases where no sensitivity to exogenous antigen can be detected), through physical exertion or the like, induced asthma, rhinitis, thrush disease, bird fancier's disease, bronchitis, cough of various kinds (including whooping cough) and such blockages in the nose and bronchi, which usually occur in conjunction with colds. The novel mixtures and compositions are also of value in the treatment of other disease states, wherein the disease is either caused by or accompanied by antigen-antibody reactions or excessive mucus secretion.

The novel compositions of the invention can be administered in a variety of ways, and they may operate systemically or locally. Thus, for example, the compounds may be administered by oral or nasal inhalation into the lung, or they may be administered directly to the nose, eye, oral cavity, esophagus, or other available body surfaces. The new mixtures may be administered directly to the organ or body part showing symptoms, or also to a body part other than the one showing the symptoms.

The novel mixtures of the present invention can be used according to many different dosing programs, either alone or in combination with one or more other active compounds.

The active components of the blends can be prepared in the form of various (particle) sizes in a very wide range.

Thus, for inhalation and other uses, the compounds may have a mass median diameter of 0.01-10 µm, preferably!

2-6 and most preferably 2-4 pm. Larger crystals or agglomerates, e.g., granules or hard pellets of the compounds - which often have greater bulk density than the finely divided (finely powdered) materials - may be used as intermediates in the preparation of the compounds' formulations, for example, tablets or they are used for their own sake, g or for filling capsules. The finely divided compounds can also be agglomerated into soft pellets or granules with, on the one hand, sufficient strength for both mechanical packaging, for example encapsulation and transport, but on the other hand sufficient fragility to be broken into fine particles.

when used in an inhalation device.

452 249

The compositions of the present invention preferably comprise less than 80% by weight and more preferably less than 50% by weight of a composition of the invention mixed with a pharmaceutically acceptable adjuvant, diluent (diluent) or carrier.

It is particularly preferred that the composition should not contain such material which may cause a harmful, for example, allergic reaction in the patient. Materials which can cause adverse reactions are described in more detail in Belgian Patent Specification 854 690.

For administration by inhalation, the novel mixtures can be prepared into preparations with either a pressurized gas, e.g., nitrogen, or a liquid-transferred propellant, to form a pressurized aerosol composition, preferably containing 1-20% by weight of the active agents. . Furthermore, the composition should preferably contain less than about 5% by weight water; even more preferably it should be substantially anhydrous.

The liquefied propellant is preferably gaseous at room temperature (20 ° C) and atmospheric pressure (760 mm Hg) and should be non-toxic. Examples of suitable liquefied propellants which may be used are, inter alia, alkanes containing up to 5 carbon atoms, e.g. butane or pentane, or a C ^-Cg alkyl chloride, e.g. methyl, ethyl or propyl chlorides. The most preferred liquid-transferred propellants are fluorinated and fluorinated chlorinated C ^-C₂- (preferably

C 1-4 alkanes, for example those marketed under the trade name Freon The preferred halogenated alkanes can generally be represented by the formula C_N]hrs_nClyF_zwherein m is an integer below 3, n is an integer or 0, y is an integer or 0, and z is an integer such that n + y + z = 2m + 2. Examples of these propellants are dichlorodifluoromethane ( Propellant 12), 1,2-dichlorotetrafluoroethane (Propellant 114), CC1F2 «CC1F2, trichloromonofluoromethane (Propellant 11), dichloromonofluoromethane (Propellant 21), monochlorodifluoromethane (Propellant 22), trichlorotrifluoroethane (Propellant 113) Mixtures of the aforementioned propellants may be used to obtain better steam pressure characteristics, e.g. Propellant 11 with Propellant 12, or Propellant '12 with Propellant 114. Preferred compositions which

452 249 'does not contain Propellant 11. It is desirable that the vapor pressure of the propellant used be between 3500 and 4550 p / cm

24 ° C.

The composition may also contain a surfactant, e.g. a liquid or solid nonionic surfactant or solid anionic surfactant.

The preferred solid anionic surfactant is sodium dioctyl sulfosuccinate.

The amount of surfactant is selected taking into account the solids content of the suspension and the particle size of the solids.

When a liquid nonionic surfactant is used, it should have a hydrophilic-lipophilic balance (HLB ratio) of less than 10 and preferably 1-5.

Preferably, the surfactant comprises 0.05-1.5% by weight of the total composition.

Suitable nonionic surfactants are phospholipids, e.g., endogenous phospholipids, esters or partial esters of, on the one hand, fatty acids having 6-22 carbon atoms, such as capric acid, octyl acid, lauric acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, oleic acid and other oleic acid. an aliphatic polyhydric alcohol or its cyclic anhydride such as ethylene glycol, glycerol, erythritol, arabitol, mannitol, sorbitol, the hexitol anhydrides derived from the sorbitol (the sorbitan esters marketed under the name of Spain), and the polyoxyethylene and polyoxypropylene derivatives of these esters. You can also use mixed esters, for example mixed or natural glycerides. The preferred liquid nonionic surfactants are the oleate of sorbitan, for example those marketed under the names Arlacel C® (sorbitan sequioleate), Span 80® (sorbitan monooleate) and Span 85® (sorbitan trioleate). Other suitable nonionic surfactants are sorbitan monolaurate, polyoxyethylene sorbitol tetraoleate, polyoxyethylene sorbitol pentaoleate, polyoxypropylene mannitiol dioleate and lecithin.

452 249

For inhalation in the form of a powder preparation, the active agent (s) in finely divided form can be used in admixture with a more coarse-grained carrier, comprising particles of e.g. up to 400 µm diameter. It is preferred that at least 90% by weight of the particulate (s) of the active agent have an effective particle size below 10 µm (preferably a particle size of 0.01 - 10 µm), that at least 90% by weight of the carrier particles here will have an effective particle size below 400 µm and that at least 50% by weight of the carrier particles have an effective particle size above 30 microns. In the case of particles below 30 microns, the effective particle size can be measured by a Coulter counter. In the case of particles above 30 microns, the effective particle size can be measured by an Alpine "air jet screen.

Preferably, at least 95% by weight of the active agent particle (s) has an effective particle size in the range of 0.01 µm. At least 90% by weight, preferably at least 95% by weight, preferably having an effective particle size of 0.1-10 µm. It is preferred that at least 50% by weight of the particles of the active agent (s) have an effective particle size in the range of 2-6 µm.

The particle size spectrum chosen for the carrier will depend on the particular inhalation device to be used for dispersing the preparation. However, it is desirable to avoid carrier particles less than 10 µm; the number of non-drug particles that penetrate deep Hedge in the lung is thus kept to a minimum. A large proportion of very large particles can result in the preparation feeling grainy ”in the mouth, which is why such preparations are less preferred. The use of a coarse particle carrier can also cause problems in filling operations with such filling machines having a metering device which takes up powder by being submerged (dipped) in a powder bed from above. In contrast, the use of a coarse particle carrier may facilitate the filling operation when using machines in which a matrix is ​​filled from above; but the large particle size can possibly. cause a tendency for the composition to segregate during transport or storage. Thus, it is desirable that at least 95% by weight of the carrier particles have an effective particle size of less than 400 µm. Preferably, at least 50% by weight, preferably at least 70% by weight of the carrier particles, should have an effective particle size in the range of 30-150, especially 30-80 µm.

452 249

The composition preferably contains 20 - 80 weight percent, especially 30 - 70 weight percent, and even more particularly 40 - 60 weight percent of total active agents, as well as 80 - 20 weight percent, especially 70-30 weight percent, and even more particularly 60 - 40 weight percent carriers.

The powder compositions can be prepared by mixing the ingredients in one or, preferably, several (e.g., 2) steps in a mixer, e.g. planetary mixer or other stirring mixer.

The carrier may be a non-toxic material of any kind which is chemically inert to the compounds and which is acceptable for inhalation or administration through the nose. Examples of carriers scm can be used include. inorganic salts, e.g. sodium chloride or calcium carbonate; organic salts, e.g. sodium tartrate or calcium lactate; organic compounds, e.g. urea or propylidone; monosaccharides, e.g. lactose, mannitol, arabinos or dextrose monohydrate; disaccharides, e.g. maltose or sucrose; polysaccharides, e.g. starches, dextrins or dextrans. In particular, lactose is preferred as a carrier, e.g. crystalline lactose.

The powder compositions are usually introduced into sealed capsules of gelatin, plastic or other material. The container in question is usually filled loosely, to less than about 80% by volume, preferably less than about 50% by volume, with the intended powder composition. t

Alternatively, the novel mixtures, when intended for inhalation, may be used in pellet or granule form, wherein the pellet ball or granule is soft, has a diameter of 10-1000, preferably 30-500 µm, and comprises an agglomerate of individual drug particles, of which at least 90% by weight have a diameter less than 10 µm.

The soft pellet ball or granule preferably has such internal cohesion (coherence) that it remains intact when filled into a container, e.g. capsule, using automatic or semi-automatic filling machines under transport and storage conditions and in fluidization inside a container of the device from which the pellet or granule material is to be discharged; but at the same time, this coherence should preferably be such that the pellet ball or granule can still be broken into particles of therapeutically effective size outside this container as it is thrown out of the container.

. 452 249

It has been found that satisfactory soft pellet beads or granules for use in insufflators of the type described in British Patent 1,182,779 (available in the market under the brand name Spinhaler) - which. operated by being influenced by human inhalation - has an average size in the range of 50-250 µm, preferably an average size in the range of 120-160 µm, and most preferably an average size of about 140 µm.

The invention is further illustrated in the examples below.

Example 1

Pressure aerosol to give sodium chromoglycate (1 mg) and 2- (tert-butylamino) -1- (4-hydroxy-3-hydroxymethylphenyl) ethanol sulfate (0.1 mg). Sodium chromoglycate (micronized, dried) 1.4420% (w / w) 2- (tert.butylamino-1- (4-hydroxy-3-hydroxy-0.1442 methylphenyl) ethanol sulfate (micronized)

Sorbitan trioleate 1.0000

Dichlorotetrafluoroethane 38.9655

Dichlorodifluoromethane · 58.4483

The dichlorodifluoromethane is cooled to -55 ° C, whereupon the sorbitan trioleate is dispersed using a high shear mixing mixer. The sodium chromoglycate and 2-tert-butylamino) -1- (4-hydroxy-3-hydroxymethylphenyl) ethanol sulfate are dispersed in the mixture and the dichlorotetrafluoroethane, which was previously cooled to -55 ° C, is added. The suspension is filled into jars, and on each jar is attached a dosing valve with 50 µl capacity.

Example 2

Pressure aerosol to provide sodium chromoglycate (1 mg) and 1- (3,5-dihydroxyphenyl) -2- (tert-butylamino) ethanol sulfate (0.25 mg).

<td>Sodium Chromoglycate (micronized, dried) 1- (3,5-dihydroxyphenyl) -2- (tert-butylamino) -</td><td>1.4286% (w / w)</td>

<td>ethanol sulfate (micronized)</td><td> 0,3571</td>

<td>trioleate</td><td> 1,0000</td>

<td>trichlorofluoromethane</td><td> 24,3036</td>

<td>dichlorotetrafluoroethane</td><td> 24,3036</td>

<td>dichlorodifluoromethane</td><td> 48,6071</td>

452 249

The sorbitan trioleate is dissolved in the trichlorofluoromethane and the sodium chromoglycate and 1- (3,5-dihydroxyphenyl) -2- (tert-butylamino) ethanol sulfate are dispersed in the solution. The solution is cooled to -55 ° C and the dichlorotetrafluoroethane and dichlorodifluoromethane are cooled to ~ 55 ° C. The suspension is filled into jars, and a dosing valve with 50 µl capacity is attached to each can. IN

Example 3

Pressure aerosol, which provides sodium chromoglycate (1 mg) and D, L-7- £ 3-Ζϊβ,

3,5-trihydroxyphenylethylamino-7-propyl) -1,3-dimethylxanthine hydrochloride t (0.5 mg).

Sodium Chromoglycate (micronized, dried) 1.4663

D, L-7- [3- (3,3,5-trihydroxyphenylethyl) - 0.7331 amino7-propyl) -1,3-dimethylxanthine hydrochloride (micronized)

Sorbitan trioleate 0.5000

Trichlorofluoromethane 10,0000

Dichlorotetrafluoroethane 13.0951

Dichlorodifluoromethane 7 * 1,2055

The preparation is carried out in the same way as in Example 2.

Example 4

Dry powder for inhalation, containing in each cartridge sodium chromoglycate (20 mg anhydrous weight), 2- (tert-butylamino) -1- (4-hydroxy-3-hydroxymethylphenyl) ethanol sulfate (0.2 mg) and sieved lactose.

Sodium chromoglycate (micronized with 8% water) 50.0? wt / wt (as wet-free material)

2- (tert-butylamino) -1'-hydroxy-3-hydroxymethyl-0.5? w / w

-phenyl) -ethanol sulfate (micronized) Lactose (sieved, 30-80 µm) qs od 100?

The 2- (tert-butylamino) -1- (4-hydroxy-3-hydroxymethylphenyl) ethanol sulfate is mixed with the sieved lactose by tearing both components together, and these components are mixed with the sodium chromoglycate using a planetary mixer. The mixture is filled into hard gelatin capsules no. 2 so that each capsule contains the desired amount of the active ingredients.

452 249

Example 5

Dry powder for inhalation containing in each cartridge the equivalent amount of 20 mg of anhydrous sodium chromoglycate and 0.5 mg of 1- (3,5-dihydroxyphenyl) -2- (tert-butylamino) ethanol sulfate in a carrier of sieved lactose.

Sodium Chromoglycate (micronized with 8 $ water) 50.0 $ w / w (as anhydrous material)₃5 (dihydroxyphenyl) -2- (tert-butylamino) - 2.5 $ w / w

ethanol sulfate (micronized)

Sieved lactose qs at 100.0 $ w / w

The preparation is carried out in the same way as in Example 4.

Example 6

Dry powder for inhalation, containing in each cartridge the equivalent amount of 20 mg of anhydrous sodium chromoglycate and 0.5 mg of 1- (3,5-dihydroxyphenyl) -1-hydroxy-2-ZX4-hydroxyphenyl) -isopropylamino7-ethane hydrobromide in a carrier of sieved lactose .

NA Sodium Chromoglycate (micronized with 8 $ water) 5θ, θ $ w / w (as anhydrous) 1- (3,5-dihydroxyphenyl) -1-hydroxy-2- (4-hydroxy) _{2 vikfc}/_weightphenyl) isopropylamino? -ethane hydrobromide (micronized)

Sieved lactose qs at 100.0 $ w / w

The preparation is carried out in the same way as in Example 4.

Example 7

Soft pellet preparation for inhalation, containing in each cartridge the equivalent amount of 20 mg of anhydrous sodium chromoglycate and 0.1 mg of 2- (tert-butylamino) -1- (4-hydroxy-3-hydroxymethylphenyl) yl ethanol sulfate. Sodium chromoglycate (micronized) 99.51% w / w (as anhydrous material)

2- (tert-butylamino) -1- (4-hydroxy-3-hydroxy-0.49 $ w / w methylphenyl) ethanol sulfate (micronized)

The sodium chromoglycate is exposed to water vapor so that its moisture content will be between 9.5 and $ 10.5 (w / w). The 2- (tert-butylamino) -1- (4-hydroxy-3-hydroxymethylphenyl) ethanol sulfate is triturated with 1/10 of the sodium chromoglycate, and the mixture so obtained is mixed with the remaining portion of the sodium chromoglycate for 15 minutes. · Drumming in a rotating drum. The mixture is screened through a screen with 40 meshes per cm, and the screened powder at 452 249

I «ΐ tumbled for another 15 min. in the rotating drum. The product fills in hard gelatin capsules no. 2 so that each capsule will contain the desired amount of the active ingredients.

Example 8

Soft pellet formulation for inhalation, containing in each cartridge the equivalent of 20 mg of anhydrous sodium chromoglycate and 0.25 mg of 1- (3-5-dihydroxyphenyl) -2- (tert-butylamino) ethanol sulfate. Sodium Chromoglycate (Micronized) 98.77? w / w (as water; ten-free material) 1- (3,5-dihydroxyphenyl) -2- (tert-butylamino) - 1.23? w / w

ethanol sulfate (micronized)

The preparation is carried out in the same way as in Example 7.

Example A

Anaphylactic bronchospasm in rats passively sensitized with anticonvulsant serum

Materials and methods

1st Preparation of the antiserum for passive sensitization *

Rats are sensitized as follows:

Day 0: Conalbumin (5 mg / kg) in plus B.pertussis vaccine (0.5 ml / animal) ip. Day 10: N. brasiliensis (4 x 10 5 larvae). Day 24 or 25: Blood is taken from the rats by cardiac puncture, the antiserum is separated and the resulting antiserum portions are pooled. . .

2nd Passive sensitization

Sprague Dawley rats are passively sensitized by injecting the antiserum prepared as above iv.

The strength of the antiserum varies from batch to batch.

Therefore, a trial should be carried out using groups of rats given 0.125 - 1.0 ml of serum doses, so as to determine the dose that provides good submaximal bronchospasm.

3rd Preparatory measures4 for recording and measuring the anaphylactic bronchoconstriction

18-24 hours. after sensitization, the rats are anesthetized with pentobarbitone. In the rat hs tail, a cannula is inserted, using a needle inside a thin tube to provide a flexible supply line for iv dosing. Then, each animal is arranged for

452 249 measurement of resistance in the airways. A cannula is inserted into the trachea and air is inflated using a Starling miniature respirator pump. This is made to operate at a stroke volume of 3-5 ml and a rate of 72 beats / min. The inlet pressure is kept constant at 9 cm water column using a water valve. At the start of the experiment, the stroke volume is set so that with each inhalation 1 air bubble will be forced out through the water valve. This overflowing air, which thus does not enter the trachea, passes through a pneumotacograph tube connected to a pressure difference transfer device (Grass pT5A) and a recording device showing changes in the air flow rate. The increase in the overflow caused by antigen-induced bronchoconstriction after injection of 25 mg / kg iv hen egg white, is measured as% of the maximum overflow (the latter being obtained by throttling the trachean tube).

4, Effect of intravenously administered test compounds

After control recording of normal breathing below 3-5 »in. a dose of the test compound or an equivalent volume of saline iv is given 1 minute before the antigen dose indicated above. The inhibitory action of the test compound or mixture is determined as

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