EP1901732A2

EP1901732A2 - Use of activators of the soluble guanylate cyclase for promoting the healing of wounds

Info

Publication number: EP1901732A2
Application number: EP06762453A
Authority: EP
Inventors: Thomas Krahn; Johannes-Peter Stasch; Gerrit Weimann; Wolfgang Thielemann; Beatrix Stelte-Ludwig
Original assignee: Bayer Healthcare AG
Current assignee: Bayer Pharma AG
Priority date: 2005-07-06
Filing date: 2006-07-06
Publication date: 2008-03-26
Also published as: CA2614083A1; WO2007003435A3; DE102005031575A1; WO2007003435A2; US20090221573A1; EP2301547A1; JP2009500364A

Abstract

The invention relates to the use of substances for producing a pharmaceutical product/medicament for promoting the healing of wounds.

Description

Use of soluble guanylate cyclase activators to promote wound healing

The present invention relates to the use of substances for the manufacture of a pharmaceutical product / medicament for promoting wound healing.

The process of wound healing or the regulation of healing of damaged tissue generally involves the recruitment of inflammatory cells, followed by the fibroblasts that accumulate around the wound. Collagen and other connective tissue constituents are deposited and rearranged to restore the original connective tissue (e.g., in the skin, muscle, ligaments, and tendons). Therefore, it is of great clinical value to find medicines that promote this process after surgery, injury or wound healing disorders. Wound healing is disturbed, i. delayed or inhibited, e.g. in patients with systemic diseases such as diabetes, kidney or liver damage, peripheral vascular disease, and in patients taking medications that interfere with wound healing, e.g. Corticosteroids, immunosuppressants and vascular growth inhibitors.

Any interruption of the anatomical or physiological function of a body tissue is called a wound. This means that not just a cut that hurts the skin is a wound. Even if an internal organ does not work properly anymore, that's a wound. Often, these physiological wounds are considered more of a violation. Therefore one differentiates between open and closed wounds. Open wounds include e.g. Cutting, stinging, locating, cracking, scraping, biting and bullet wounds as well as skin detachment and traumatic amputations. Closed wounds include e.g. Bruises, bruises, distortions, burns, frostbites, chemical wounds from acids or alkalis, actinic wounds and necrosis (e.g., tissue damage due to poor circulation, e.g., heart and brain infarcts).

The aim of wound healing is complete regeneration, ie to restore the normal state of the tissue. However, the regeneration processes are not always able to reach this state. Therefore, it is often only an incomplete regeneration, for example, when deeper skin layers (eg dermis) are affected. Affected are also tissues that can hardly renew (heart and nerve tissue). Here, the lost tissue is usually filled by functionally inferior replacement tissue (connective tissue), which is less perfused and less elastic and functional than the original tissue. This is called a scar or scar tissue. To the maximum Thus, retrieving functionality may also be a goal to promote functional tissue (eg, muscle cells) in its regeneration against connective tissue (fibroblasts).

It is known that mechanisms (for example NO-releasing substances) which lead to an increase in the intracellular messenger cGMP can also contribute to an improvement in wound healing (WO 96/08966-A1, Murrell).

The activation (caused by agonists) of the soluble guanylate cyclase leads to an increase of the intracellular messenger cGMP. Surprisingly, it has now been found that the activators of soluble guanylate cyclase according to the invention, compounds I-VI, which are listed below, are suitable for the production of pharmaceutical substances / medicaments for promoting wound healing, in particular in humans.

Compound (I) corresponds to the following formula:

Compound (I), its preparation and use as a medicament have already been disclosed in WO 01/19780.

Compound (II) corresponds to the following formula:

Compound (II), its preparation and use as a medicament have already been disclosed in WO 00/06569.

Compound (III) corresponds to the following formula:

Compound (HI), their preparation and use as medicaments have already been disclosed in WO 00/06569 and WO 02/42301.

Connection (FV) corresponds to the following formula:

Compound (FV), their preparation and use as medicaments have already been disclosed in WO 00/06569 and WO 03/095451.

Connection (TVa) corresponds to the following formula:

Compound (FVa), their preparation and use as medicaments have already been disclosed in WO 00/06569 and WO 03/095451.

Compound (V) corresponds to the following formula:

Compound (VI) corresponds to the following formula:

Compounds (V) and (VI), their preparation and use as medicaments have already been disclosed in WO00 / 02851.

The present invention is the use of compounds of formulas (I-VI) and their salts, hydrates, hydrates of the salts for the preparation of a medicament for the promotion of wound healing.

Another object of the present invention is the pharmaceutical composition containing at least one compound of formulas (I-VI) with a pharmaceutically acceptable carrier, each for each of the therapeutic effects previously discussed. The composition may be used alone or in combination with at least one other substance, e.g. a stabilizing substance.

The compounds according to the invention can act systemically and / or locally. For this purpose they may be suitably applied _^ be, for example oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, dermal, transdermal (topical), conjunctival, otic or as implant or stent. For these administration routes, the compounds according to the invention can be administered in suitable administration forms.

For the oral administration are according to the prior art functioning, rapidly and / or modified compounds of the invention donating application forms containing the compounds of the invention in crystalline and / or amorphized and / or dissolved form, such. Tablets (uncoated or coated tablets, for example with enteric or delayed-release or insoluble coatings which control the release of the compound of the invention), orally disintegrating tablets or films / wafers, films / lyophilisates, capsules (for example hard or soft gelatin capsules) in the oral cavity , Dragees, granules, pellets, powders, emulsions, suspensions, aerosols or solutions.

Parenteral administration can be accomplished by bypassing a resorption step (e.g., intravenously, intraarterially, intracardially, intraspinal, or intralumbar) or by resorting to absorption (e.g., intramuscularly, subcutaneously, intracutaneously, percutaneously, or intraperitoneally). For parenteral administration are suitable as application forms u.a. Injection and infusion preparations in the form of solutions, suspensions, emulsions, lyophilisates or sterile powders.

For the other routes of administration are suitable, for example Inhalation medicaments (including powder inhalers, nebulizers), nasal drops, solutions or sprays, lingual, sublingual or buccal tablets, films / wafers or capsules, suppositories, ear or ophthalmic preparations, vaginal capsules, aqueous suspensions (lotions, shake mixtures), lipophilic suspensions, ointments, creams, transdermal therapeutic systems (such as patches), milk, pastes, foams, scattering powders, implants or stents.

The compounds according to the invention can be converted into the stated administration forms. This can be done in a conventional manner by mixing with inert, non-toxic, pharmaceutically suitable excipients. These excipients include, but are not limited to, excipients (e.g., microcrystalline cellulose, lactose, mannitol), solvents (eg, liquid polyethylene glycols), emulsifiers and dispersing or wetting agents (e.g., sodium dodecyl sulfate, polyoxysorbitanoleate), binders (e.g., polyvinylpyrrolidone), synthetic and natural polymers (e.g., albumin). , Stabilizers (eg antioxidants such as ascorbic acid), dyes (eg inorganic pigments such as iron oxides) and flavor and / or odoriferous agents. Another object of the present invention are pharmaceutical compositions containing at least one compound of the formula (I-VI) of the invention, usually together with one or more inert, non-toxic, pharmaceutically suitable excipients, and their use for the purposes mentioned above.

In general, it has been found to be beneficial to administer amounts of about 0.01 to 5000 mg / kg, preferably about 0.5 to 1000 mg / kg, of body weight per day to achieve effective results.

Nevertheless, it may nevertheless be necessary to deviate from the stated amounts, depending on body weight, route of administration, individual behavior towards the active ingredient, type of preparation and time or interval at which administration takes place. Thus, in some cases it may be sufficient to manage with less than the aforementioned minimum amount, while in other cases, the said upper limit must be exceeded. In the case of the application of larger quantities, it may be advisable to distribute these in several single doses throughout the day.

The preferred subject of the present invention is the topical application of a pharmaceutical formulation.

Another object of the present invention are suspensions and ointments for

• topical application containing at least one of the compounds of formulas (I-VI). Other topical formulations of the invention include solutions, sprays, lotions, gels, creams, powders, powder sprays, pastes, emulsions, foams, and sticks containing at least one

Active ingredient of the formula (I-VI), optionally also contain several active ingredients.

The topical application of an active ingredient of formula (I-VI) also takes place in the form of patches, spray patches, occlusive dressings, envelopes and controlled delivery systems. In these preparations, the active ingredients may be contained in dissolved or suspended form.

Ointments contain as a basis hydrocarbon gels, lipogels, absorption bases, W / O ointment bases, mixed emulsions or polyethylene glycols.

Creams contain O / W bases.

Pastes contain, in addition to an ointment or cream base, high proportions of powdery constituents such as, for example, zinc oxide, talc, starch or titanium dioxide. GeIe contain solvents such as water, ethanol, isopropanol or propylene glycol and are prepared using gelling agents such as cellulose ethers, alginates, polyacrylates, bentonite, gelatin, tragacanth, polyvinylpyrrolidone or polyvinyl alcohol. It is also possible to use lipophilic gel bases or microemulsions.

Powders contain powdery additives such as starch, stearates, silicon dioxide, clay, magnesium carbonate, talc, cellulose, zinc oxide and in particular lactose.

Stabilizers, antioxidants, preservatives, humectants, moisturizers, solvents or adjuvants to enhance penetration and efficacy may be added to all formulations.

Examples of penetration enhancers are propylene glycol, polyethylene glycol, dimethylsulfoxide, decylmethylsulfoxide, azone, N-methylpyrrolidone, diethyltoluamide, ethanol, isopropyl myristate, isopropyl palmitate, oleic acid and its esters, medium chain triglycerides, dimethylisosorbitol, 2-octyldodecanol, branched chain fatty acid esters, benzyl alcohol, urea, salicylates and surfactants.

As another pharmaceutical formulation suitable for topical application, a liposome-based carrier system known in the art from pharmaceutical technology can be used. This particular should be the penetration of

Improve lipids in the epidermis. Such liposome dispersions with different

Inclusion compounds and phospholipids as bilayer membrane-forming agents or so-called empty liposomes without inclusion compounds have been described in numerous publications and have already been clinically tested. The preparation takes place e.g. by treatment of an aqueous phospholipid dispersion with ultrasound, dispersion of phospholipids with surfactants in aqueous phase, dissolution of phospholipids in organic solvents, removal of the

Solvent by lyophilization and dispersion of the residue in the aqueous phase,

Infusion methods or reverse phase evaporation. Preferred pharmaceutical formulations for topical application are finely dispersed systems based on lipid mixtures.

From document WO 96/37192 pharmaceutical or cosmetic compositions are known which in combination with a sphingolipid or glycolipid (ceramide) comprise: a partial fatty acid ester of polyoxyethylene sorbitan, a phospholipid, a triglyceride and a therapeutic agent, in water (and possibly alkanol) as a carrier liquid. From EP 0852941 compositions are known in which the active ingredients ^' by a similar excipient mixture, the type of partial fatty acid ester of polyoxyethylene sorbitan, Phospholipid and ethanol, even in water-insoluble, liquid to highly viscous, oily or at least oil-soluble active ingredients, such as fat-soluble vitamins, therapeutic oils or sunscreen substances, in the form of a dispersion of nanoparticles (nanodispersion) solubilize. The pharmaceutical formulations containing compounds of the formulas (I-VI) described in this disclosure are considered as preferred subject of the present invention.

The formulations may contain, according to the procedure, active substance between 0.1 and 99% active ingredient, suitably 25-95% for tablets and capsules and 1-50% for liquid formulations, i. the active substance should be present in sufficient quantities to reach the stated dose latitude.

Another object of the present invention is the use of a combination of one or more of the compounds according to the invention (I-VI) with one or more other substances. Suitable combination partners are, for example, substances that promote vessel growth. By way of example and preference, cGMP-increasing substances, adenosine agonists and growth factors are mentioned in this context.

Another object of the present invention is the use of a combination of one or more of the compounds of the invention (I-VI) with one or more substances selected from the group of medicaments for promoting blood circulation, pain relief or antibiotics.

Experimental part:

1. Influence of the migration of smooth muscle cells in vitro by administration of a soluble guanylate cyclase activator:

Human coronary artery smooth muscle cells are seeded on a 6-well plate (1.5 × 10 ⁵ cells / well) (Clonetics) and cultured for 48 h in SmGM2 medium (Clonetics). Before the

Using the plates, the wells are coated with vitronectin (50 ng / cm ² ; Gibco BRL). Half of the confluent cell monolayer is wounded using a cell scraper. In the cell-free area, at least 50% of the vitronectin coating is retained. Unless otherwise stated, the culture medium is replaced by MCDB-131 / 0.2% BSA (Gibco / BRL) medium for the experiment.

The test substances are added in various concentrations and the migration distance into the cell-free area is monitored by means of a microscope for 24 h and 48 h. Each measuring point represents the mean of four measured regions. PDGF, as a potent chemotactic factor for SMC, is used as a positive control (1 nM, 10 nM, R & D Systems). To inhibit integrin-induced migration, the RGD tripeptide (Bachern) is used. In order to determine the antagonistic activity of the test substances, the cells are stimulated in the presence of the substance with 1 nM or 10 nM PDGF and the migration distances are determined in correlation to the PDGF-induced migration distances.

Representative results from this test are shown in the following table:

Table. Inhibition of horizontal migration of human smooth muscle cells

Coronary artery (48 h incubation period)

The compounds I-VI show that soluble guanylate cyclase activators are suitable for promoting wound healing. 2. Formulation of the active ingredients

Ophthalmic topical application

The active compounds are used to form a 2 wt .-% suspension Liquifilm ^® Eye Drops (Allergan, Ettlingen, Germany) (ml containing polyvinyl alcohol 14 mg 1, as well as mg as excipient chlorobutanol x V2 H ₂ O 5.25 sodium chloride) and the mixture in a glass tube Sonicated for 15 min in an ultrasonic bath at 15 ⁰ C.

Dermatological topical application

I) Alcoholic suspension

The active compounds are mixed to form a 2% by weight suspension with 30% by weight of isopropyl myristate / 70% by weight of ethanol and sonicated in a glass tube for 15 minutes in an ultrasonic bath at 15 ° C.

2} Spreadable preparations

Wool wax alcohol ointment according to DAB 9 is used to prepare the ointment preparation. The active ingredient is suspended in a molten mixture of 0.5 parts by weight of cetylstearyl alcohol (quality according to DAB 9), 6 parts by weight of wool wax alcohol (quality according to DAB 9) and 93.5 parts by weight of white Vaseline (quality according to DAB 9). The mixture is stirred until it cools to room temperature (about 21 ⁰ C).

To produce a so-called "PEG-Isogel" preparation, the active ingredient is suspended or dissolved in the melt of a mixture of low-molecular-weight polyethylene glycol (PEG) and higher molecular weight PEG After suspending or dissolving the active ingredient in the melt, the preparation is heated until the Cool to room temperature.

Low molecular weight PEG is liquid at room temperature (about 21 ⁰ C), higher molecular weight PEG is solid at room temperature (cuttable, waxy).

The mixing ratios of the two PEG types depend on the required viscosity of the preparation and on the average molecular weights of the PEG types used.

As examples, preparations of 6.5 parts by weight of PEG 400 and 1 part by weight of PEG 4000 or 7 parts by weight of PEG 400 and 2 parts by weight of PEG 6000 can be used. Mixing ranges are from 1: 1 to 10: 1 (in each case parts by weight of low molecular weight PEG to higher molecular weight PEG). Preferably, the range is from 2: 1 to 8: 1, more preferably in the range from 3: 1 to 7: 1.

As a further gel preparation is a polyacrylate gel (so-called "Carbogel"), which consists of 1 part by weight Carbopol 974 P NF (manufacturer Fa. BF Goodrich, USA), 5 parts by weight of isopropyl alcohol, 5 parts by weight of sodium hydroxide (5 wt .-%) and 89 parts by weight of water To prepare the preparation, the Carbopol is rubbed with the isopropanol, the active ingredient is added and the mixture is dispersed in water, the sodium hydroxide solution is gradually added with stirring and a gel is formed.

Claims

claims

1. Use of compounds of the formulas (I-VI)

(IVa)

and their salts, hydrates, hydrates of the salts for the preparation of a medicament for promoting wound healing.

2. Use according to claim 1, wherein the medicament is used for a topical dosage form.

Use according to claim 1, wherein the medicament is given subcutaneously or intramuscularly.

4. Use according to any one of claims 1 to 3, wherein the medicament is used preventively.

5. Use of any one of claims 1 to 4, wherein the medicament is for treatment of wounds ^• averaging is applied in diabetics invention.

6. A pharmaceutical composition for the promotion of wound healing, which contains at least one substance according to claim 1.

7. A pharmaceutical composition according to claim 6, which additionally contains a medicament for promoting blood circulation, analgesia or antibiotics.