EP2538948A1

EP2538948A1 - Biodegradable material containing silicon, for pro-angiogenetic therapy

Info

Publication number: EP2538948A1
Application number: EP11706209A
Authority: EP
Inventors: Iwer Baecker; Christoph Suschek; Magda Ulrich; Bouke Boekema
Original assignee: Bayer Innovation GmbH
Current assignee: Bayer Innovation GmbH
Priority date: 2010-02-24
Filing date: 2011-02-22
Publication date: 2013-01-02
Also published as: WO2011104215A1; US20130115187A1; BR112012021355A2; JP5992341B2; CA2790610A1; MX2012009717A; CA2790610C; KR20130036005A; JP2013520463A; CN102834101B; RU2573989C9; CN102834101A; DE102010008981A1; AU2011219897A1; RU2012140381A; RU2573989C2

Abstract

The invention relates to a biodegradable material containing silicon, for the prophylaxis and/or treatment of diseases involving reduced and/or disturbed angiogenesis and/or diseases for which an increased angiogenesis rate is required for the healing process.

Description

Silicon-containing, biodegradable material for pro-angiogenic therapy

The present invention relates to a silicon-containing, biodegradable material for the prophylaxis and / or treatment of diseases associated with diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis is conducive to the healing process.

Angiogenesis refers to the growth of small blood vessels (capillaries), predominantly by budding from a preformed capillary system. It is a complex process in which the endothelial cells, pericytes and smooth muscle cells required to form the vessel walls are activated by various angiogenic growth factors such as Fibroblast Growth Factor (FGF) and Vascular Endothelial Growth Factor (VEGF). Angiogenesis is of considerable biological and medical importance. In modern medicine, two forms of therapeutic application of the principle of angiogenesis are distinguished: anti- angiogenic therapy and pro- angiogenetic therapy. Pro-angiogenic protein therapy uses growth factors with angiogenic potency, most notably Fibroblast Growth Factor 1 (FGF-1) and Vascular Endothelial Growth Factor (VEGF); With these growth factors, the greatest clinical experience is available. But also the growth factors epidermal growth factor (EGF), platelet-derived endothelial cell growth factor (PD-ECGF) and platelet-derived growth factor (PDGF) and transforming growth factor (TGF) possess a certain angiogenic potency. The experience so far in clinical trials, in particular with FGF-1, is promising: new blood vessels in the human myocardium as well as a myocardial perfusion improvement (along with an increase in patient stress) have been demonstrated.

Silicon is a trace element that is important for humans in bonded silicate form. Silicon is a building block of those proteins that are responsible for the strength and elasticity of the tissues. It is also incorporated in connective tissue, bones, skin, hair, nails and blood vessels. In addition, silicon strengthens the immune system of the body, the so-called immune system, and promotes wound healing. Deficiency in silicon results in stunted growth, loss of bone stability with increased risk of osteoporosis, premature hair loss, brittle nails, and skin changes. Possible changes of the skin are increased wrinkling, dryness, scaling, increased horn formation, itching, thickening and painful, slit-shaped tears of the skin due to reduced elasticity. In addition, the immune system of the body, the so-called immune system, through the Silicon deficiency weakened and there is an increased susceptibility to infections. Although silicon-containing compounds have been described for the prophylaxis or for the treatment of some diseases. However, it has not been known to date that compounds containing silicon can also induce or promote angiogenic processes and are therefore suitable for pro-angiogenic therapies.

US2006 / 0178268A1 describes an aqueous solution consisting of non-colloidal silica and boric acid for the treatment of bone, cartilage, skin, arteries, connective tissue, joint, hair, nail, skin diseases and osteoporosis, rheumatic diseases, arteriosclerosis , Arthritis, cardiovascular diseases, allergic diseases and degenerative diseases.

US2006 / 0099276A1 discloses a method for producing a silicic acid derivative by hydrolysis of a silicone compound to oligomers in the co-presence of a quaternary ammonium compound, an amino acid or an amino acid source or combinations thereof. The silica extrudate can be used as pharmaceuticals for the treatment of infections, nails, hair, skin, tooth, collagen, connective tissue, bone diseases, osteopenia, cell formation for degenerative (aging) processes. US6,335,457B 1 discloses a solid wherein silica is complexed with a polypeptide. This patent also discloses therapeutically useful mixtures containing this solid.

WO2009 / 018356A1 relates to a mixture comprising a sodium phosphate compound, an ammonium compound and a silicate for the prophylaxis or treatment of diseases such as prostate cancer, colorectal cancer, lung cancer, breast cancer, liver cancer, neuronal cancer, bone cancer, HIV syndrome, rheumatoid arthritis, multiple sclerosis, Epstein Barr virus, fibromyalgia, chronic fatigue syndrome, diabetes, Bechets syndrome, irritable bowel syndrome, Crohn's disease, bedsores, trophic ulcers, radiation- or chemotherapy-weakened immune systems, hematomas or combinations thereof.

WO2009 / 052090A2 describes a method for the treatment of inflammatory diseases, autoimmune diseases, bacterial or viral infections and cancer by using a composition containing silicate. US2003 / 0018011A1 relates to a pharmaceutical composition comprising a fatty acid and a water-soluble silicate polymer as anti-allergic or anti-inflammatory agent.

No. 5,534,509 relates to a pharmaceutical composition comprising a water-soluble silicate polymer as the active agent with a saccharide or sugar alcohol as the inert carrier substance for the treatment of allergies, inflammation, pain or for the improvement of peripheral blood circulation or paraesthesia.

DE19609551C1 describes the production of bioabsorbable (continuous) fibers based on polyhydroxysilicic acid ethyl ester. The fibers are used as reinforcing fibers for biodegradable and / or bioabsorbable (implant) materials. The fibers can also be used for the production of biodegradable composite materials.

WO01 / 42428A1 describes a method for the production of a skin implant in which skin cells are applied to the surface of a nutrient solution and these are grown using a surface element consisting of the fibers described in DE19609551C1.

EP1262542A2 relates to a process for the production of cells, tissues and organs, wherein a fiber matrix is used as cell support substance and / or lead structure according to DE19609551 C1.

WO2006 / 069567A2 relates to a multilayer dressing in which a fiber matrix according to DE19609551C1 is also used in one layer. The multi-layer dressing can be used for the treatment of wound defects such as chronic diabetic-neuropathic ulcers, leg ulcers, bedsores, secondary healing infected wounds, non-irritating, primary healing wounds such as, in particular, ablative lacerations or abrasions.

WO2008 / 086970A1, WO2008148384A1, PCT / EP2008 / 010412 and PCT / EP2009 / 004806 describe inter alia the preparation of further polyhydroxy-silicic acid ethyl ester compounds which can be used according to the invention. The compounds are generally described for use as bioabsorbable materials in human medicine, medical technology, filter technology, biotechnology or the insulation industry. It is also mentioned that the materials can be advantageously used in the field of wound treatment and wound healing. Fibers can be used, for example, as surgical sutures or as reinforcing fibers. Nonwovens can be superficial in the supply Wounds, in filtration of body fluids (eg blood) or in the bioreactors used as Anzuchthilfe.

It is not disclosed in the prior art that the above-mentioned biodegradable polyhydroxy-silicic acid ethyl ester compounds (eg in the form of a fiber or a nonwoven) are used for the prophylaxis and / or treatment of diseases associated with a diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis is conducive to the healing process. Although the above-cited documents describe the use of the polyhydroxy-silicic acid ethyl ester compounds for wound treatment and wound healing, it is known that wound healing is associated with pro- geniogenetic processes, but the prior art does not disclose that the above-mentioned biodegradable Polyhydroxysilicic acid-ethyl ester compounds can generally be used for a pro-angiogenetische therapy. This is surprising also in view of the fact that it has not been described so far for other silicon-containing compounds that they can be used for a pro-angiogenic therapy.

The present invention therefore relates to a silicon-containing, biodegradable material for the prophylaxis and / or treatment of diseases associated with a diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis is conducive to the healing process, wherein the silicon-containing , biodegradable material is a polyhydroxysilicic acid ethyl ester compound, with the proviso that the wound defects, such as chronic diabetic neuropathic ulcer, ulcus cruris, decubitus wounds, secondary healing infected wounds, irritating, primarily healing wounds, such as ablative lacerations or abrasions are excluded. The invention also encompasses the use of a silicon-containing, biodegradable polyhydroxysilicic acid ethyl ester compound according to the invention for the preparation of a medicament for the prophylaxis and / or treatment of diseases associated with diminished and / or impaired angiogenesis and / or diseases for the an increased rate of angiogenesis is conducive to the healing process, with the proviso that the wound defects, such as chronic diabetic neuropathic ulcer, ulcus cruris, bedsores wounds, secondarily healing infected wounds, irritating, primarily healing wounds, such as in particular ablative lacerations or abrasions are excluded.

Excluded from the invention are those applications of the material according to the invention which are described in the following patents DE19609551 C1, WO01 / 42428A1, EP 1 262542A2, WO 2006/069567A2, WO2008 / 086970A1, WO2008,148384A1, PCT / EP2008 / 010412 and PCT / EP2009 / 004806 and those associated with neo-angiogenesis. The use of a polyhydroxysilicic acid ethyl ester fiber fleece as a constituent of a multi-layer dressing is described in WO2006 / 069567A2 for the treatment of wound defects such as chronic diabetic-neuropathic ulcer, leg ulcers, bedsores wounds, secondarily healing infected wounds, non-irritating, primarily healing wounds, in particular ablative lacerations or abrasions Service. EP1262542A2 describes a wide variety of tissue engineering applications of polyhydroxysilicic acid ethyl ester compounds according to the invention. The term "tissue engineering applications" according to the present invention is based on the products, processes and applications described in EP1262542A2, except for the tissue engineering applications of the silicon-containing and biodegradable material according to the invention, if these are discussed in EP1262542A2 be associated with a pro-angiogenic therapy.

The term "" Polyhydroxykieselsäureethylester- compound "refers to compounds of the general formula H [OSi ₈ 0i ₂ (OH) _x (OC ₂ H ₅₎ _6-x] _n OH, wherein x is 2 to 5 and n> 1 ( Polymer). The inventive silicon-containing, biodegradable material is preferably present as a material in the form of a fiber, a fiber matrix, as a powder, as a monolith and / or as a coating. Such a silicon-containing, biodegradable material can be prepared according to the invention as described below:

a) at least one hydrolysis-condensation reaction of tetraethoxysilane,

b) evaporation to produce a single-phase solution, preferably with simultaneous gentle mixing of the reaction system,

c) cooling of the single-phase solution and

d) ripening to produce a silica sol material

e) drawing threads from the silica sol material to generate a fiber or a

Fiber matrix and / or drying and in particular spray or freeze-drying of the silica sol material for generating a powder and, if necessary, dissolving the powder in a solvent to generate a liquid formulation and / or coating an article to be coated with the silicon-containing, biodegradable material with the Silica sol material, and / or casting the silica sol material into a monolith generating mold.

According to the invention, the silicon-containing, biodegradable material of the invention preferably exists as fiber, fiber matrix (fleece), as powder, as liquid formulation and / or as coating. In a further embodiment of the invention, the subject silicon-containing, biodegradable material is prepared as described above, wherein the tetraethoxysilane in step a) at an initial pH of 0 to <7, optionally in the presence of a water-soluble solvent, preferably ethanol, in a Temperature is acid catalysed from 0 ° C to 80 ° C and in step b) evaporating to a single-phase solution having a viscosity in the range of 0.5 to 2 Pa · s at a shear rate of 10 s ^"1 at 4 ° C. is carried out.

In a further embodiment of the invention, the silicon-containing, biodegradable material is prepared as described above, wherein the acid catalysis in step a) with nitric acid H ₂ 0 in a molar ratio to Si compound in the range 1: 1, 7 to 1: 1 9, preferably in the range of 1: 1, 7 to 1: 1.8. The hydrolysis-condensation reaction in step a) is preferably carried out at a temperature of 20 to 60 ° C, preferably 20 to 50 ° C over a period of at least one hour. Preferably, the hydrolysis-condensation reaction proceeds in step a) over a period of several hours, such as 8 hours or 16 hours. However, this reaction can also be carried out over a period of 4 weeks. Step (b) is preferred in a preferred embodiment of the invention in a closed apparatus in which mixing is possible (preferably rotary evaporator or stirred tank) with simultaneous removal of the solvent (water, ethanol) by evaporation at a pressure of 1 to 1013 mbar at a pressure of <600 mbar, optionally with continuous feed of a chemically inert towing gas for partial pressure reduction of the evaporating components of 1 - 8 m ³ / h (preferably at 2.5 to 4.5 m ³ / h), a reaction temperature of 30 ° C to 90 ° C, preferably 60 to 75 ° C, more preferably at 60 to 70 ° C and preferably with gentle mixing of the reaction system to 80U / min (preferably at 20U / min to 80U / min) up to a viscosity of the mixture 0.5 to 30 Pa · s at a shear rate of 10 s ^-1 at 4 ° C., preferably 0.5 to 2 Pa · s at a shear rate of 10 s ^-1 at 4 ° C., particularly preferably about 1 Pa s (measurement at 4 ° C, shear rate 10 s ^-1 ). In a further embodiment of the invention, the silicon-containing, biodegradable material in step c) to preferably 2 ° C to 4 ° C, cooled. At this low temperature, the maturing preferably takes place (step d). Maturation may take several hours or days to about 3 to 4 weeks. The ripening process in step d) is preferably carried out up to a viscosity of the sol of 30 to 100 Pa · s at a shear rate of 10 s ^-1 at 4 ° C. and a loss factor of 2 to 5 (at 4 ° C., 10 1 / s , 1% deformation). The drawing of threads from the silica sol material in step e) is preferably carried out via a spinning process. Such a spinning process step can be carried out under customary conditions, as described, for example, in DE 196 09 551 C1 and DE 10 2004 063 599 A1. In a preferred embodiment of the invention, the pressure during the spinning of the silica sol material is selected so that a throughput of at least 80 g / h based on the total sol throughput is achieved.

Preferably, the spun fibers are exposed directly after spinning for a period of 30 to 60 minutes to the same climatic conditions as in the spinning tower (i.e., e.g., -19% air humidity, ~25 ° C temperature). This step is referred to below as conditioning. The fibers obtained by this process are called conditioned fibers.

In another preferred embodiment, the conditioned fibers are exposed to a relative humidity of at least 35% (at room temperature) for a period of 1 to 30 minutes and preferably a period of 1 to 10 minutes prior to use (see also Table 2).

The drying of the silica sol material to generate powder is preferably carried out by spray or freeze drying. A powder can also be obtained by comminuting and grinding monoliths or else fibers according to the invention. To generate a liquid formulation, the powder is dissolved in a solvent. Suitable solvents may be aqueous or oily depending on the application (e.g., injection solution or suspensions). The coating of a material to be coated with the silicon-containing, biodegradable material with the silica sol material is preferably carried out by immersing the body to be coated in the silica sol, by casting or by spin coating or spraying the silica sol. The silica sol material according to step d) can also - in order to generate a monolith - be poured into a mold and then dried.

Further details regarding the preparation of the inventive silicon-containing, biodegradable materials are DE19609551C1, WO01 / 42428A1, EP 1262542A2, WO2006 / 069567A2, WO2008 / 086970A1, WO2008148384A1, PCT / EP2008 / 010412 and PCT / EP2009 / 004806.

For the purposes of the present inventions, the term "biodegradable" refers to degrading the property of the polyhydroxy-silicic acid ethyl ester compound of the present invention when the material is exposed to conditions typical of those present in a tissue regeneration (e.g., a wound) degradable or biodegradable in the invention is in particular the polyhydroxy-silicic acid ethyl ester compound according to the invention if, after 48 hours, preferably 36 hours and more preferably after 24 hours, in a 0.05 M Tris pH 7.4 buffer solution ( Fluka 93371) thermostatically dissolved at 37 ° C.

The term "diseases associated with diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis promotes the healing process" describes all those diseases that can be treated (or prevented) by pro-angiogenic therapy Such diseases include:

Diseases of the blood circulation as well as the cardiovascular system such as:

Anemia, angina pectoris, (pheripheral) vascular disease, arteriosclerosis, Winiwarter-Bürger disease, myocardial infarction, ischemia especially of the heart muscle, lung, cardiomyopathy, congestive heart failure, coronary artery diseases such as coronary restenosis, viable haemorrhagic telangiectasis, hypercholesterolemia, ischemic heart disease, myocardial scleroderma, myointimal hyperplasia, clogged blood vessels, peripheral atherosclerotic vascular disease, portal hypertension, preeclampsia, rheumatic heart disease, hypertension, thromboembolic diseases,

Bone cartilage or muscle-associated diseases such as:

Bone / cartilage repair, bone defect, bone fracture, bone growth, cartilage disease, bank disc degeneration, osteoarthritis, osteoporosis, spinal fracture, fibromyalgia, polymyositis,

Diseases of the central nervous system such as:

Central nervous system or peripheral nervous system ischemia, Alzheimer's, amyotrophic lateral sclerosis, autonomic neuropathy, aneurysms, cerebral infarction, stroke, cerebrovascular disease, cerebrovascular deficiency, dementia, epilepsy, ischemic peripheral neuropathy, mild cognitive deficits, multiple sclerosis, Nerve damage, Parkinson's disease, Niemann-Pick disease, polyneuropathy,

Schizophrenia, spinal cord injury, toxic neuropathy;

d) diseases of the eye such as:

glaucoma; retinopathy;

e) gastro-intestinal diseases such as:

Crohn's disease, gastric ulcer, indestinal ischemia, irritable bowel syndrome, pancreatitis,

Ulcerative colitis;

f) Hormonal or metabolic diseases such as:

Diabetes mellitus, diabetic foot, peripheral diabetic vascular disease;

g) diseases of the immune system such as:

Allergies, mastocytosis, Sjögren's disease, transplantation rejection, tissue defects in collagenoses such as Sjögren's syndrome, dermatomyositis, systemic lupus

Erythematosus, CREST syndrome, Sharp syndrome;

h) Infectious diseases such as:

Septic shock

i) kidney diseases like:

Nephropathy, intracranial hypertension, renal ischemia;

j) oral diseases such as:

Dental plaque, gum disease,

k) diseases of the reproductive system such as:

erectile dysfunction,

1) Respiratory diseases such as:

Asthma, bronchopulmonary dysplasia, pneumonia, respiratory distress syndrome, m) disorders of the skin such as:

Nonspecific dermatitis, decubitus ulcers, dermal ischemia, dermal ulcers, diabetic gangrene, diabetic skin ulcers, lacerations, psoriasis, scleroderma, skin lesions, burns, surgical wounds, wound healing

n) Vascular diseases such as:

Vascular insufficiency, vascular restenosis, vasculitis, vascular spasm, Wegener's granulomatosis

o) Other diseases like:

Hair loss, lactic acidosis, body limb ischemia, cirrhosis of the liver, hepatic ischemia, mitochondrial encephalomyopathy, sarcoidosis, soft tissue defects (in particular by accident, surgery or malformation), diseases treated by autografts of tissues and / or organs. The term "diseases associated with diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis promotes the healing process" in a preferred embodiment describes diseases selected from the following group:

a) Diseases of the circulatory system and the cardiovascular system such as:

Ischemia, in particular of the heart muscle;

b) Central nervous system diseases such as:

Ischemia in the central nervous system or in the peripheral nervous system.

c) oral diseases such as:

Dental plaque, gum disease.

d) soft tissue defects, diseases treated by autografts of tissues and / or organs.

An object of the invention also relates to the use of inventive (r) silicon-containing, biodegradable materials with auto-transplants for the treatment of diseases which are treated by autografts of tissues and / or organs. In this procedure, a silicon-containing, biodegradable material according to the invention is used in addition to the autograft in order to achieve an improved angiogenesis and thus a faster incorporation and a better acceptance of the autologous transplant into the existing tissue.

A further preferred subject matter of the invention relates as silicon-containing, biodegradable material to a polyhydroxysilicic acid ethyl ester compound having an ethoxy group content of at least 20%, preferably of at least 25% and more preferably of 25 to 30%. Preferably, a polyhydroxysilicic acid ethyl ester compound having such an ethoxy group content is in the form of a fiber or a fiber matrix.

The ethoxy group content is measured by the known standard Zeisel ether cleavage method after spinning within a period of 1 to 4 weeks after spinning, the polyhydroxysilicic acid ethyl ester compound being reduced in humidity (ie, for example, within a package of absorbents such as in European Patent Application EP09007271). A further preferred subject matter of the invention relates to a polyhydroxysilicic acid ethyl ester compound in the form of a fiber or a fiber matrix in which the fiber or the fiber matrix has a compressibility of at least 17%, preferably 20% and particularly preferably at least 25%, as the silicon-containing, biodegradable material ,

The compressibility is measured by the following method steps:

a) measurement of the thickness of the polyhydroxysilicic acid ethyl ester compound in the form of a fiber matrix at at least two different pressures,

b) Plotting the measured value pairs (measured thickness and pressure) in a diagram as thickness over log (pressure),

c) Regression according to (d / do) = (p / p ₀ ) ^"b , where p _{D is} a pressure of 0.25 kPa, cL is the calculated thickness of a fiber matrix at p ₀ and b is the exponent of the curve is

d) calculation of the compressibility [k] on the basis of a regression according to k = (1 - in the di _; 25 corresponds to the thickness calculated from the regression for 1.25 Pa.

The compressibility is measured within a period of one week after spinning, with the polyhydroxy-silicic acid ethyl ester compound being stored at reduced humidity (i.e., within an absorbent package, for example) during the time before the measurement.

The appropriate dosage of the polyhydroxy silicic acid ethyl ester compound is generally between 0.001 to 100 mg / kg body weight per day and administered in single or multiple doses. Preferably, a dosage between 0.01 and 25 mg / kg, more preferably 0.1 to 5 mg / kg per day is used. The biodegradable properties of the polyhydroxy-silicic acid ethyl ester compounds, however, also make it possible to apply the compounds in higher dosages and, for example, subcutaneously degrade them within the body as depots in the form of a monolith over a longer period of time and promote pro-angiogenic processes. The material according to the invention or a precursor thereof (such as the silica sol material described above in step d)) can be processed with the commonly used in galenics carriers, fillers, Zerfallbeeinflussern, binders, lubricants, absorbents, diluents, Geschmackskorrigentien, colorants, etc. and converted into the desired application form. Reference may be made to Remington's Pharmaceutical Science, 15th ed. Mack Publishing Company, East Pennsylvania (1980). The material of the present invention may be administered orally, mucosally (such as sublingually, buccally, rectally, nasally or vaginally), parenterally (such as subcutaneously, intramuscularly, by bolus injection, intraarterial, intravenous), transdermally, or locally (such as, for example, direct application on the skin or topical application to an exposed organ or wound).

In particular, tablets, dragees, film-coated tablets, capsules, pills, powders, granules, lozenges, suspensions, emulsions or solutions are suitable for oral administration.

For example, as described above, tablets, dragees, capsules, etc. may be obtained by casting the silica sol material obtained in step d) into a tablet-like or capsule-like form for generating a monolith. However, the tablets and capsules can also be prepared by the above-described inventive material in the form of a powder by conventional methods. The material according to the invention or a precursor thereof can be prepared with known excipients, for example inert diluents such as dextrose, sugar, sorbitol, mannitol, polyvinylpyrrolidone, disintegrants such as corn starch or alginic acid, binders such as starch or gelatin, lubricants such as magnesium stearate or talc and / or agents for achieving a depot effect such as carboxylpolymethylene, carboxymethylcellulose, cellulose acetate phthalate or polyvinyl acetate are added. Tablets can also consist of several layers. The capsules containing the subject matter of the invention can be prepared, for example, by mixing the materials according to the invention or a precursor thereof with an inert carrier such as lactose or sorbitol and encapsulating them in gelatine capsules. Coated tablets can accordingly be prepared by coating cores produced analogously to the tablets with agents customarily used in tablet coatings, for example polyvinylpyrrolidone or shellac, gum arabic, talc, titanium oxide or sugar. In this case, the dragee wrapper can also consist of several layers, wherein the auxiliaries mentioned above in the case of the tablets can be used. For parenteral administration, injection and infusion preparations are possible. For intraarticular injection, appropriately prepared crystal suspensions may be used. For intramuscular injection, liquid formulations such as aqueous and oily injection solutions or suspensions and corresponding depot preparations can be used. For rectal administration, the materials according to the invention can be used in the form of suppositories, capsules, solutions (for example in the form of enemas) and ointments both for the systemic and for the Local therapy can be used. Furthermore, may be mentioned as a preparation and agents for vaginal use. Liquid formulations such as injection solutions or suspensions can be obtained, for example, by adding the above-described material according to the invention in the form of a powder with suitable aqueous or oily solvents. Other types of preparation are known to the person skilled in the art. Solutions or suspensions of the material according to the invention may additionally contain taste-improving agents such as saccharin, cyclamate or sugar and, for example, flavorings such as vanillin or orange extract. They may also contain suspending aids such as sodium carboxymethylcellulose or preservatives such as p-hydroxybenzoates. Suitable suppositories can be prepared, for example, by mixing with suitable carriers such as neutral fats or polyethylene glycol or derivatives thereof. The solutions described can also be used, for example, for the treatment of dental plaque or gum disease (eg via an injection or for rinsing the oral cavity). For transdermal application, patches or formulations for topical application in gels, ointments, fatty ointments, creams, pastes, powders, milk and tinctures are possible. Wound plasters are preferably made of fibers or a fiber matrix (fleece) of the materials according to the invention as described in the prior art. In a further embodiment of the invention, the material according to the invention or a precursor thereof can be produced by a coating method, for example by dipping an article or body to be coated in the silica sol material described above in step d), by casting or by spin-coating or spraying a be coated such silica sol material. The silica sol material is preferably applied to implants, autografts, vascular prostheses, dental prostheses or heart valves, and particularly preferably to autografts, dental prostheses and heart valves.

The abovementioned forms of application may also contain other pharmaceutical active substances which can be added during the production process.

Legend

FIG. 1: Neoangiogenesis with the addition of VEGF and material according to the invention (PKEE =

Polyhydroxykieselsäureethylester- connection) in the form of a fiber matrix to human endothelial cells (in vitro) detected via specific antibodies against the surface marker CD31. Control shows neo-angiogenesis of human endothelial cells without addition of VEGF or PKEE (negative control).

Neo-angiogenesis with the addition of VEGF and material according to the invention (PKEE = polyhydroxysilicic acid ethyl ester compound) in the form of a fiber matrix (nonwoven) to human endothelial cells (in vitro) detected via specific antibodies against von Willebrand factor (vWF). K = negative control.

Quantitative evaluation of the neo-angiogenesis of VEGF and material according to the invention in the form of a fiber matrix (fleece) in human endothelial cells. K = negative control; S / CD31 = material according to the invention and detection of neoangiogenesis via CD31 antibodies; S / vWF = material according to the invention and detection of neoangiogenesis via vWF antibody; V / CD31 = VEGF and detection of neoangiogenesis via CD 31 antibody; V / vWF = VEGF and detection of neoangiogenesis via vWF antibody. * = p <0.05 vs control (Student t-test).

Quantitative evaluation of neoangiogenesis of VEGF (V) and material according to the invention in the form of a fiber matrix (S / Tl = fiber matrix type I; S / T2 = fiber matrix type II; S / T3 = fiber matrix type III; S / T4 = fiber matrix type IV , see Preparation Ex. 1) in human endothelial cells, taking into account the staining of the microvessels with an anti-vWF antibody. K = negative control; * = p <0.05 vs control (Student t-test).

Quantitative evaluation of the neo-angiogenesis of VEGF (V) and of the material according to the invention in the form of a fiber matrix (S / T; S / T2; S / T3; S / T4 = type I, type II, type IV, fibrous matrix) in human Endothelial cells taking into account the staining of microvessels with an anti-CD31 antibody. K = negative control; * = p <0.05 vs control (Student t-test).

Quantitative analysis of the VEGF concentration in cell culture supernatants of human endothelial cells in the absence (control = K) and presence of different materials according to the invention in the form of a fiber matrix (nonwoven; (S / T; S / T2; S / T3; S / T4 = fibrous matrix type I) Type II, Type III, Type IV)); # = p <0.05 compared to the control and fiber matrix type II to IV (anova tukey test); * = p <0.05 compared to control (Student t-test).

Quantitative analysis of the influence of surmarin on neo-angiogenesis in human endothelial cells, taking into account the staining of the microvessels with an anti-CD31 antibody in a control (K = only human endothelial cells, K + Su = human endothelial cells and addition of surmarin), when added of the material according to the invention (Si = only material according to the invention and Si + Su = material according to the invention and Surmarin) and addition of VEGF (V = addition of VEGF and V + Su = addition of VEGF and Surmarin). # = p <0.05 compared to control (anova tukey test), * = p <0.05 compared to cultures without surmarine (Student t-test); Figure 7 shows that the materials of the present invention induce angiogenesis via VEGF. In the presence of the material (Si) according to the invention, it is possible to observe an increase of approximately 3.5 times (compared to the control, approximately 350%) of the percentage area of microvessels. With Surmarin, this effect can be reversed.

Examples

1. Preparation of Inventive Fiber Matrices from Polyhydroxysilicic Acid Ethyl Ester 1124.98 g of TEOS (tetraethoxysilane) were placed in a stirred tank as starting material for the hydrolysis-condensation reaction. 313.60 g of EtOH as solvent were added. The mixture is stirred. Separately, 1N HNO ₃ (55.62g) was diluted with H ₂ O (120.76g) and added to the TEOS-ethanol mixture. The mixture was stirred for 18 hours. The mixture obtained by this step was subsequently evaporated at temperatures of 62 ° C with the addition of a trailing stream and stirring (60 rpm) to a dynamic viscosity (shear rate 10 s-1 at 4 ° C) of 1 Pa · s.

The solution was then quiescent in a closed polypropylene rag and kept upright at a temperature of 4 ° C to a dynamic viscosity of about 55 Pa · s (shear rate 10 s-1 at 4 ° C) and a loss factor of 3.0 matured.

The sol resulting from the ripening was then spun into the fiber. The fibers were produced in a conventional spinning plant. For this purpose, the dope was filled into a pressure cylinder cooled to -15 ° C. The dope was pressed through the nozzles with pressure. The flowable, honey-like material fell by its own weight in a spin shaft located under the pressure cylinder with 2 m length. Temperature and humidity were controlled in the spinning shaft. The temperature was 25 ° C and the humidity at 19%. When the threads hit the traversing table, they almost retained their cylindrical shape, but were still so fluid that they glued together at their contact surfaces to form Fasliegegen (nonwovens). The fibers thus spun are exposed directly after spinning for a period of 35 minutes to the same climatic conditions as in the spinning tower (ie for example a humidity of 19%, a temperature of 25 ° C.) (conditioning of the spun fibers). A total of 8 different fiber webs of polyhydroxysilicic acid ethyl ester (type I to IV, AI, A2, Bl and B2) were prepared. The spun fibers have a diameter of about 50 μιη. The fiber webs AI, A2, B1 and B2 differ in terms of a different throughput in spinning (and thus the spinning time, see Table 1). The throughput indicated in g / h in Table 1 refers to the total sol flow rate. The pressure in the spinning container is adjusted so that the desired throughput is achieved. Type I to IV AI A2 Bl B2

Throughput approx. 80 g / h approx. 57 g / h approx. 99 g / h approx. 58 g / h approx. 97 g / h

Spinning time 6,15 min 8,5 min 5,25 min 12,2 min 7,8 min

per fleece (5

cm x 5 cm)

Table 1: Throughput and spinning time of fiber matrices according to the invention

Polyhydroxykieselsäureethylester

The fibrous webs Type I, Type II, Type III and Type IV differ in that, after the conditioning step described above and the packaging of the nonwovens, they are exposed to an environment with a relative humidity of 35% to 55% for storage for different periods until they are used During storage of the nonwovens in the packaging, the humidity in the packaging is greatly reduced by the presence of absorbents. Suitable packaging for the storage of fiber webs can be found for example in European Patent Application EP09007271.

Different production of fiber matrices according to the invention from polyhydroxysilicic acid ethyl ester. During the specified period, the fleeces are exposed to the following environment: temperature: 25 ° C; Humidity 35% to 55% Mass Thickness of Compressibility Ethoxy Dressing Group

salary

Type i 420 mg 1.7 mm 21% 26.1%

Type II 390 mg 1.7 mm 16% 17.3%

Type III 380 mg 1.7 mm 15% 12.7%

Type IV 365 mg 1.7 mm 13% 6.6%

AI 436 mg 2.0 mm 26% 26.8%

A2 419 mg 1.5 mm 17% 26.8%

Bl 622 mg 2.8 mm 26% 26.8%

B2 620 mg 2.0 mm 15% 26.8%

Table 3: Different product properties of inventive fiber matrices

Polyhydroxykieselsäureethylester

The different production conditions led to different nonwoven properties, in particular with regard to the compressibility and the ethoxy content of the nonwovens after spinning (see Table 3).

The compressibility was measured and calculated by thickness measurements (Precision Thickness Gauge Model 2000, Wolf Messtechnik GmbH) on the basis of the method steps described in the description. The ethoxy group content was measured by the standard Zeisel ether cleavage method. An internal standard solution was added to a fibrous matrix to be analyzed and, after addition of hydriodic acid, heated for one hour at 120 ° C. in a gas-tight glass vessel for one hour. Existing ethoxy groups are converted into ethyl iodide. The resulting ethyl iodide is determined by gas chromatography, the evaluation is carried out according to the method of the internal standard. The standard is toluene. 2. Neoangiogenesis in human endothelial cells

Experimental setup: To determine neoangiogenesis, the angiogenesis assay kit from TCS Cellworks (Buckingham, UK) was used. 24-well cell culture plates were used whose soil is confluent with a cell lawn consisting of human fibroblasts and human endothelial cells. All cell culture media necessary for the procedure as well as antibodies for the detection of endothelial-specific surface antigens (CD31, Von Willebrand Factor = vWF) were also purchased from TCS Cellworks. For carrying out the experiment, suitable plastic hooks were also used for 24-well cell culture plates, which can be loaded with different substrates. The content of the Kunststoffeinhänge is separated by a membrane of the medium of cell culture. Due to the membrane permeability, however, a mass transfer of dissolved substances between the contents of the suspensions and the cell culture medium is possible.

Experimental procedure: The cells in the openings of the culture plate to be examined were covered with 300 μl cell culture medium per opening. Subsequently, all openings were equipped with the Kunststoffeinhängen. In the case of testing the polyhydroxysilicic acid ethyl ester compound, in each case 1 cm ^{2 of} a polyhydroxysilicic acid ethyl ester fiber matrix was introduced into the suspension and covered with 350 μl medium. In controls, the inserts were supplemented with 350 .mu.l medium, in the positive controls the inserts were supplemented with 350 .mu.l medium + 2 ng / ml VEGF and in the negative controls were the suspensions with 350 .mu.l medium + 20 ug / ml suramin, a potent VEGF- Inhibitor, filled up. The culture plates were cultured for 7-12 days, with full or partial replacement of the medium or medium contents every three days. In the quantitative analysis of the effect of surmarin on neoangiogenesis on human endothelial cells, shown in Figure 7, surmarin was co-administered with the polyhydroxy-silicic acid ethyl ester fiber matrix (see Si + Su) or VEGF (see V + Su). Evaluation: To determine the rate of angiogenesis, after 7 to 12 days of culture, the suspensions and media were removed from the cell culture plates and the confluent cells fixed on the bottom of the cell culture plate according to the manufacturer's instructions. To illustrate the formation of microvessels, the fixed preparations were stained using the endothelial cell-specific antibodies according to the manufacturer's instructions. Similarly, using a VEGF ELISA kit (R & D Systems, Abingdon, UK) determined the concentration of VEGF in all supernatants of the experiments.

Results: After staining the respective cultures with endothelial cell-specific antibodies it became apparent that in cultures incubated with the polyhydroxysilicic acid ethyl ester fiber matrix, the density of microvascular formation both after staining with CD31 (FIG. 1) and after staining with the vWF-specific antibody (FIG. Figure 2) was higher than in the untreated controls and comparable or higher to microvascularization in the VEGF-containing positive controls.

For the quantitative determination of the observations mentioned, the digital images of the test results were densitometrically evaluated with the aid of the image processing software "ImageJ". As shown in Figures 3-5, the relative vascular density in samples treated with polyhydroxy-silicic acid ethyl ester fiber matrices is significantly higher than in the control cultures and comparable to cultures maintained in the presence of VEGF.

To analyze the effect of polyhydroxysilicic acid ethyl ester fiber matrices on endothelial VEGF synthesis, the above-mentioned experiments were extended to 12 days and the culture medium was not changed to maintain cell vitality, but extended to fresh medium every 4 days. As a result, the accumulated amount of VEGF in the whole experiment could be recorded and thus VEGF levels well above the detection limit of the test could be recorded. As shown in Figure 6, the incubation of endothelial cells with polyhydroxy-silicic acid ethyl ester fiber matrices resulted in significantly increased VEGF synthesis in the assay approach, with type I polyhydroxy-silica ester polymer matrices inducing a significantly increased VEGF production compared to the other types of silicates.

The term "vessel density" refers to the area covered by newly formed capillary structures in the culture plate in relation to the total area The measurement of the vessel density is carried out by densitometric determination of the black pixel portions in a black-and-white image of the capillary structures colored by specific antibodies in comparison to that white surface of the plate background without capillary structures.

The term "percentage area of microvessels" describes what percentage of the empty area (control corresponds to 100%) is taken up by neoangiogenesis-induced microvessels. The measurement of this parameter was done densitometrically. We examined black and white photos of the cultures for their percentage of black pixels (^ positive antibody staining of the endothelial cells). 3. In vivo experiments on neo-angiogenesis of the material according to the invention

Polyhydroxy-silicic acid ethyl ester fiber matrices (AI, A2, B1 and B2) according to the invention were prepared in an animal model (Schwein; Middelkoop E, et al., Porcine wound models for skin substitution and burn treatment. Biomaterials. 2004 Apr; 25 (9): 1559-67). was compared with the clinical gold standard (nSHT = reticulate split skin graft, MDM = Matriderm® by Dr. Suwelack Skin & Health Care AG.). In Yorkshire pigs 3 x 3 cm and 2.7 mm deep wounds (open wounds 3rd degree) were created. On these open wounds, the polyhydroxy-silicic acid ethyl ester fiber matrices (A I, A 2, B 1 and B 2) according to the invention and the contours were transplanted and compared with each other. Each matrix was applied to 4 different wounds. 13 days after transplantation biopsies were taken from the wound area and immunohistochemistry was performed. Woundbrand Factor (vWF; Ulrich MM, et al., Wound Repair Rain, 2007 Jul-Aug., 1997) was described in relation to the blood vessels ; 15 (4): 482-90) stained with an antibody. The staining was evaluated by digital image analysis. The NIS-Ar software (Nikon) was used to quantify the results. Significantly increased staining (approximately 2.8 fold) of vWF regions from the material of the invention was observed as compared to the control (see Table 4).

nSHT MDM A1 A2 B1 B2 Table 4: Percent staining of vWF in biopsies on day 13 post-transplant. * Significantly different from nSHT (MWU test, * = p <0.05)

Claims

claims

A silicon-containing, biodegradable material for the prophylaxis and / or treatment of diseases associated with decreased and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis is conducive to the healing process, wherein the silicon-containing, biodegradable material is a polyhydroxy-silicic acid ethyl ester compound is, with the proviso the wound defects, such as chronic diabetic neuropathic ulcer, ulcus cruris, decubitus wounds, secondary healing infected wounds, irritating, primarily healing wounds, such as in particular ablative tear or abrasions are excluded.

Silicon-containing, biodegradable material according to claim 1, wherein the material is in the form of a fiber, a fiber matrix, as a powder, as a liquid formulation, as a monolith and / or as a coating.

Silicon-containing, biodegradable material according to claim 2, produced by: a) at least one hydrolysis-condensation reaction of tetraethoxysilane b) evaporation to produce a single-phase solution, preferably with simultaneous gentle mixing of the reaction system,

c) cooling of the single-phase solution and

d) ripening to produce the silica sol material

e) drawing threads from the silica sol material to generate a fiber or a fiber matrix and / or drying, in particular spray drying or freeze drying the silica sol material to generate a powder and optionally. Dissolving the powder in a solvent to generate a liquid formulation and or

Coating an article to be coated with the silicon-containing biodegradable material with the silica sol material; and / or casting the silica sol material into a mold to generate a monolith. 4. The silicon-containing, biodegradable material prepared according to claim 3, wherein the tetraethoxysilane in step a) at an initial pH of 0 to <7, optionally in the presence of a water-soluble solvent, preferably ethanol, at a temperature of 0 ° C to 80 ° C acid catalyzed and

in step b) by evaporating a single-phase solution to a viscosity in the range of 0.5 to 2 Pa · s at a shear rate of 10 s 1 at 4 ° C is performed. Silicon-containing, biodegradable material according to claim 4, wherein the acid catalysis in step a) with nitric acid H ₂ 0 in a molar ratio to the Si compound in the range 1: 1.7 to 1: 1.9, preferably in the range of 1: 1.7 to 1: 1.8 is performed.

A silicon-containing, biodegradable material for the prophylaxis and / or treatment of diseases associated with diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis is conducive to the healing process of any one of claims 1 to 5, wherein the diseases associated with diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis is conducive to the healing process are selected from the group of: a) circulatory and / or cardiovascular diseases such as:

Anemia, angina pectoris, (pheripheral) vascular disease, arteriosclerosis, Winiwarter-Bürger disease, myocardial infarction, ischemia, especially of the heart muscle, lung, cardiomyopathy, congestive heart failure, coronary artery diseases such as coronary restenosis, traumatic haemorrhagic telangiectasis, hypercholesterolemia, ischemic heart disease, myocardial scleroderma, myointimal hyperplasia, clogged blood vessels, peripheral atherosclerotic vascular disease, portal hypertension, preeclampsia, rheumatic heart disease, hypertension, thromboembolic diseases,

b) Bone cartilage or muscle associated diseases such as:

Bone / cartilage repair, Bone defect, Bone fracture, Bone growth, Cartilage disease, B anks cheib endeg eneration, Osteoarthritis, Osteoporosis, Febrile fracture, Fibromyalgia, Polymyositis,

c) central nervous system diseases such as:

Central nervous system or peripheral nervous system ischemia, Alzheimer's, amyotrophic lateral sclerosis, autonomic neuropathy, aneurysms, cerebral infarction, stroke, cerebrovascular disease, cerebrovascular deficiency, dementia, epilepsy, ischemic peripheral neuropathy, mild cognitive deficits, multiple sclerosis, nerve damage, Parkinson's disease, Niemann's disease Pick disease, polyneuropathy, schizophrenia, spinal cord injury, toxic neuropathy;

d) diseases of the eye such as:

glaucoma; retinopathy;

e) gastro-intestinal diseases such as: Crohn's disease, gastric ulcer, indestinal ischemia, irritable bowel syndrome, pancreatitis, ulcerative colitis;

f) Hormonal or metabolic diseases such as:

Diabetes mellitus, diabetic foot, peripheral diabetic vascular disease;

g) diseases of the immune system such as:

Allergies, Mastocytosis, Sjögren's disease, Transplant rejection, Tissue defects in collagenoses such as Sjögren's syndrome, Dermatomyositis, Systemic lupus erythematosus, CREST syndrome, Sharp's syndrome.

h) Infectious diseases such as:

Septic shock

i) kidney diseases like:

Nephropathy, intracranial hypertension, renal ischemia;

j) oral diseases such as:

Dental plaque, gum disease,

k) diseases of the reproductive system such as:

erectile dysfunction,

1) Respiratory diseases such as:

Non-specific dermatitis, decubitus ulcers, dermal ischemia, dermal ulcers, diabetic gangrene, diabetic skin ulcers, lacerations, psoriasis, scleroderma,

Skin injuries, burns, surgical wounds, wound healing

n) Vascular diseases such as:

o) Other diseases like:

Hair loss, lactic acidosis, body limb ischemia, liver cirrhosis, liver ischemia,

Mitochondrial encephalomyopathy, sarcoidosis, soft tissue defects, diseases by

Autografts of tissues and / or organs are treated.

7. A silicon-containing, biodegradable material for the prophylaxis and / or treatment of diseases associated with a diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis is conducive to the healing process, according to one of claims 1 to 6, wherein the polyhydroxysilicic acid ethyl ester compound has an ethoxy group content of at least 20%. Silicon-containing, biodegradable material for prophylaxis and / or for

The treatment of diseases associated with diminished and / or impaired angiogenesis and / or diseases for which an increased rate of angiogenesis is conducive to the healing process of any one of claims 1 to 7 wherein the polyhydroxy-silicic acid ethyl ester compound is in the form of a fiber and / or fiber Fiber matrix is present and the fiber and / or fiber matrix has a compressibility of at least 17%.