WO1998022114A1 - Procede visant a promouvoir la reparation tissulaire - Google Patents

Procede visant a promouvoir la reparation tissulaire Download PDF

Info

Publication number
WO1998022114A1
WO1998022114A1 PCT/DK1997/000525 DK9700525W WO9822114A1 WO 1998022114 A1 WO1998022114 A1 WO 1998022114A1 DK 9700525 W DK9700525 W DK 9700525W WO 9822114 A1 WO9822114 A1 WO 9822114A1
Authority
WO
WIPO (PCT)
Prior art keywords
combination
compound
chitosan
acid
combination according
Prior art date
Application number
PCT/DK1997/000525
Other languages
English (en)
Inventor
Thorsten Jorgensen
Judi MØSS
Henrik Vigan Nicolajsen
Lise Sylvest Nielsen
Original Assignee
Dumex-Alpharma A/S
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dumex-Alpharma A/S filed Critical Dumex-Alpharma A/S
Priority to AU49412/97A priority Critical patent/AU4941297A/en
Publication of WO1998022114A1 publication Critical patent/WO1998022114A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/42—Use of materials characterised by their function or physical properties
    • A61L15/44—Medicaments
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7004—Monosaccharides having only carbon, hydrogen and oxygen atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1617—Organic compounds, e.g. phospholipids, fats
    • A61K9/1623—Sugars or sugar alcohols, e.g. lactose; Derivatives thereof; Homeopathic globules
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
    • A61K9/7007—Drug-containing films, membranes or sheets
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents
    • A61L2300/414—Growth factors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/43—Hormones, e.g. dexamethasone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/45—Mixtures of two or more drugs, e.g. synergistic mixtures

Definitions

  • the present invention relates to a combination of a first compound of an oligo- or polysaccharide containing aminosugar units and a second compound of a sulfated mono-, di- or oligosaccharide, and most preferably to a combination of chitosan and sucrose octasulfate, as an agent for the enhancement of the healing of wounds in collagen containing tissues, including skin and bone and in mucosa.
  • wounds and/or ulcers are normally found on the skin or on mucosal surfaces.
  • skin relates to the outermost surface of the body of an animal such as a human and embraces intact or almost intact skin as well as injured skin surfaces.
  • mucosa relates to undamaged or damaged mucosa of an animal such as a human and may be the oral, buccal, aural, nasal, lung, gastrointestinal, vaginal, or rectal mucosa.
  • skin is used in a very broad sense embracing the epidermal layer of the skin and - in those cases where the skin surface is more or less injured - also the dermal layer of the skin. Apart from the stratum corneum, the epidermal layer of the skin is the outer (epithelial) layer and the deeper connective tissue layer of the skin is called the dermis. The skin may have a thick or a thin epidermis and is therefore often classified as thick or thin skin. In the present context, the term "skin" embraces thick skin as well as thin skin.
  • Thick skin is found on the palms of the hands and the soles of the feet, whereas thin skin covers the remainder of the body.
  • the skin on the palms of the hands and the soles of the feet has a thick epidermis with a particularly thick layer of keratin on its outer surface.
  • the skin covering the remainder of the body has a relatively thin epidermis and the outer keratinized layer of the epidermis is relatively thin.
  • the skin forms a barrier between the body and the environment and one of the most important functions is to protect the body from invasion by potentially hazardous materials and organisms.
  • the skin consists of two layers of completely different kinds of tissue that are attached to one other over their whole extent.
  • the outer layer contains stratified squamous keratinizing epithelium. It does not contain any blood vessels so it must be nourished via tissue fluid from a second and deeper layer of the skin. This second layer contains irregularly arranged connective tissue containing blood vessels.
  • stratum corneum The outermost layer of the skin is the so-called stratum corneum. Then there is a viable layer called the epidermis and the papillary dermis layer.
  • the epidermis and particularly its layer of keratin is a barrier to disease organisms. Keratin is nearly water-proof and makes it possible to have a bath without swelling or shrinking of the skin.
  • the epidermis contains cells producing melamine and is therefore able to protect the body from harmful effects of e.g. ultraviolet light.
  • the skin serves as a temperature regulator and contains nerve endings responsible for picking up stimuli that evoke many different kinds of sensation during consciousness (e.g. touch, pressure, heat, cold and pain). Hence the skin is of great importance in permitting man to adjust to his environment.
  • the skin Since the skin is the most exposed part of the body, it is particularly susceptible to various kinds of injuries such as, e.g., ruptures, cuts, abrasions, burns and frostbites or injuries arising from various diseases. Furthermore, much skin is often destroyed in accidents. However, due to the important barrier and physiologic function of the skin, the integrity of the skin is important to the well-being of the individual, and any breach or rupture represents a threat that must be met by the body in order to protect its continued existence.
  • injuries may also be present in all kinds of tissues and injuries like abscesses and internal ulcers are also treatable with a combination according to the invention. These injuries can have a wide range of severity and can have numerous internal as well as external causes. Probably the most frequent tissue injury is skin wounds due to simple external trauma. A tissue injury on a body surface caused by a disease is typically called an ulcer irrespective of its presence on the skin, in the gastrointestinal tract on any other mucosal surface (cf. below).
  • a tissue injury starts a cascade of reactions aimed at minimising the injury and preparing for tissue repair.
  • a process that aims at miminizing the injury is typically an inflammatory reaction, which even to some extent by itself can cause tissue injury.
  • the transition from an inflammatory process to a tissue repair process is gradual and both processes can occur at the same time.
  • the tissue repair process will only start if the tissue injury process is successfully stopped either by a physiologic process alone or in combination with a specific therapy.
  • conditions in the tissue must allow that the repair process can occur. For instance in crural leg ulcers a certain minimal blood supply is required for the healing process to be possible. When these conditions are met, the tissue repair process can start.
  • tissue repair In the early stage of the tissue repair, one process is almost always involved and that is the formation of a transient connective tissue in the area of tissue injury. This process starts by forming a new extracellular collagen matrix by fibroblast. This new extracellular collagen matrix is then the support for a connective tissue during the final healing process.
  • the final healing is in most tissues a scar formation containing connective tissue.
  • tissues which have regenerative properties such as, e.g., skin and bone
  • the final healing includes regeneration of the original tissue. This regenerated tissue has frequently also some scar characteristics, e.g. a thickening of a healed bone fracture.
  • tissue repair processes include the early connective tissue formation, a stimulation of this and the subsequent processes are contemplated to improve tissue healing.
  • all tissue repair can benefit from a stimulation but the clinical impact of a stimulation of tissue healing will differ depending on the type of tissue injury and the subsequent healing process.
  • the value of a tissue healing stimulation is larger in conditions where the healing time is prolonged. A clinically important example of such a condition is leg ulcers.
  • the healing process is exemplified by description of the process for a wound present on the skin.
  • the body provides mechanisms for healing injured skin in order to restore the integrity of the skin barrier.
  • the repair process for even minor ruptures or wounds may take a period of time extending from hours and days to weeks.
  • the healing may persist for extended period of time, i.e. months or even years.
  • failure to heal of cutaneous ulcers is a serious clinical problem which is accompanied by significant financial and emotional costs.
  • the healing process is a complex and well orchestrated physiological process that involves migration, proliferation and differentiation of a variety of cell types as well as synthesis of matrix components. Normally, the healing process comprises of three phases:
  • platelets When platelets are present outside the circulatory system and exposed to thrombin and collagen, they become activated and they aggregate. Thus, platelets initiate the repair process by aggregating and forming a temporary plug to ensure haemostasis and prevent invasion from bacteria.
  • the activated platelets initiate the coagulation system and release growth factors like platelet-derived growth factor (PDGF) and epidermal growth factor (EGF).
  • PDGF platelet-derived growth factor
  • EGF epidermal growth factor
  • the first cells to invade the wound area are neutrophils followed by monocytes which are activated by macrophages.
  • neutrophils The major role of neutrophils appears to be clearing the wound of or defending the wound against contaminating bacteria and to improve the healing of the wound by removing dead cells and platelets.
  • the infiltration of neutrophils ceases within about the first 48 hours provided that no bacterial contamination is present in the wound.
  • Excess neutrophils are phagocytosed by tissue macrophages recruited from the circulating pool of blood-borne monocytes. Macrophages are believed to be essential for efficient wound healing in that they also are responsible for phagocytosis of pathogenic organisms and a clearing up of tissue debris. Furthermore, they release numerous factors involved in subsequent events of the healing process. The macrophages attack fibroblasts which start the production of collagen.
  • fibroblasts begin to proliferate and migrate into the wound space from the connective tissue at the wound edge.
  • the fibroblasts produce collagens and glycosaminoglycans and inter alia low oxygen tension at the wound stimulates proliferation of endothelial cells.
  • the endothelial cells give rise to the formation of a new capillary network.
  • Collagenases and plasminogen activators are secreted from keratinocytes. If the wound is left undisturbed and well-nourished with oxygen and nutrients, keratinocytes will migrate over the wound. Keratinocytes are believed only to migrate over viable tissue and, accordingly, the keratinocytes migrate into the area below the dead tissue and the crust of the wound.
  • the wound area is further decreased by contraction.
  • the rate of healing is influenced by the wound's freedom from infection, the general health of the individual, presence of foreign bodies, etc. Some pathologic conditions like infection, maceration, drying out, generally bad health and malnutrition can lead to formation of a chronic ulcer.
  • any procedure that can influence the rate of wound healing or favourably influence the healing of wounds is of great value.
  • Numerous treatments are currently used to create an optimal environment for the wound healing process.
  • none of these treatments have been designed to directly stimulate the wound repair process.
  • growth factors like epidermal growth factor (EGF), transforming growth factor- ⁇ (TGF- ⁇ ), platelet derived growth factor (PDGF), fibroblast growth factors (FGFs) including acidic fibroblast growth factor ( ⁇ -FGF) and basic fibroblast growth factor ( ⁇ -FGF), transforming growth factor- ⁇ (TGF- ⁇ ) and insulin like growth factors (IGF-1 and IGF-2) are conductors of the wound healing process.
  • the present invention is based on a novel approach to wound healing, namely an approach involving the use of a wound healing composition comprising as an active wound healing substance a combination of two substances, where the first substance in the combination inter alia has a function of immobilizing the combination in the wound area in order to obtain an enhanced and/or sustained or prolonged effect, and the second substance is able to bind and/or activate and/or stabilize growth factors involved in the healing process.
  • a particularly interesting combination according to the present invention is a combination between chitosan and sucrose octasulfate.
  • the combination according to the present invention has an unexpected useful, superior wound healing effect compared to the wound healing effect obtained by use of the individual substances alone or in a physical blend. Furthermore, the use of a combination according to the invention with the purpose of healing wounds is very advantageous; thus, in the case of chitosan-sucrose octasulfate the chitosan aids in immobilizing sucrose octasulfate (i.e. a wound healing agent) site specifically at/on the location at/on which the wound healing agent is to exert its effect. Moreover, the combination of e.g.
  • chitosan-sucrose octasulfate has a water solubility which is much lower than that of sucrose octasulfate by itself. If sucrose octasulfate is applied onto a wound, the pronounced water solubility of sucrose octasulfate will result in a very fast transport of sucrose octasulfate away from the site of action and, thus markedly reduce the presence of the active substance and, consequently, markedly reduce the time period during which the active substance can exert an effect.
  • the overall effect is therefore a short lasting effect of sucrose octasulfate and, accordingly, sucrose octasulfate has to be applied onto the wound very frequent (e.g. 3-7 times daily) in order to achieve the desired effect.
  • sucrose octasulfate as a wound healing agent.
  • the solubility of the sucrose octasulfate released at the desired site (wound) has been found to be of the same order of magnitude as that of sucrose octasulfate in itself.
  • chitosan in itself has also been found to have a positive impact on wound healing.
  • sucrose octasulfate and chitosan may react in situ after application of the two individual substances in vivo on a wound and form a combination of chitosan and sucrose octasulfate (this reaction takes place when the individual substances are applied in the form of powders, two different gels or in the form of solutions or two separate formulations) such a reaction proceeds in an uncontrollable manner, i.e. resulting in a combination having an uncontrollable and variable content of chitosan and sucrose octasulfate, respectively.
  • the therapeutic and/or prophylactic activity of a combination according to the invention may of course be evidenced by in vivo tests using experimental animals or humans.
  • an indication of the efficacy and/or activity of a combination according to the invention can be obtained by performing relatively simple in vitro tests such as, e.g., tests involving cell cultures.
  • a test identifying wound healing associated indicators such as, e.g., indicators of pH, partial pressure 0 2 , temperature, radical mechanisms or biotechnological assays, e.g. indicating a formation of collagen,
  • FCS fetal calf serum
  • xiii) a test involving basic fibroblast growth factor stimulation of epidermal wound healing in e.g. pigs (cf. P.A. Hebda et al., J. Invest. Dermatol. (1990), Vol. 95, 626-631),
  • xiv an in vitro test involving growing of a cell culture for the determination of the influence of the extracellular matrix on fibroblast responsiveness (cf. P.G. Genever, Br. J. Dermatol. (1995), Vol. 133, 231-235), and xv) a test involving histological examination of the tissue or cell culture.
  • the invention in a first aspect relates to a combination of a first compound of an oligo- or polysaccharide containing aminosugar units and a second compound of a sulfated mono-, di- or oligosaccharide.
  • oligosaccharide is intended to mean a carbohydrate containing from two and up to ten simple sugar (i.e. monosaccharide sugar) units linked together. Beyond ten they are denoted polysaccharides.
  • polysaccharides Within the scope of the present invention is not combinations of e.g.
  • EP-A-0 427 190 and EP-A-0 454 044 both in the name of Hoechst Aktiengesellschaft
  • polyelectrolytic complexes of e.g. chitosan and dextransulfate or xylanpolysulfate are clearly without the scope of the present invention as the complexes are composed of two polysaccharide substances.
  • the combination of the first and second substance is not merely a physical blend of the two substances since some kind of binding forces are involved in the formation of the combination. Most likely, the combination is held together by ionic binding between the amino groups of the first substance and the sulfated groups of the second substance, but other binding forces may also have an impact.
  • the combination may be a salt, a complex such as, e.g., an inclusion complex, or the substances may be held together in a polymeric network or as a mechanical inclusion, or even - although more unlikely - the substances may be covalently bound together in the combination. Further experiments are presently ongoing in order to determine the structure of the combination.
  • thermogram obtained by subjecting the combination to Differential Scanning
  • Calorimetry must be significantly different from thermograms obtained by subjecting the first compound, the second compound and a physical blend thereof, respectively, to Differential Scanning Calorimetry;
  • an IR-spectrum of the combination must be different from the spectre of the individual first and second compounds and from a physical blend of the individual first and second compounds;
  • an NMR-spectrum of the combination must be different from the spectre of the individual first and second compounds and from a physical blend of the individual first and second compounds;
  • an X-ray diffraction spectrum of the combination must be different from the spectre of the individual first and second compounds and from a physical blend of the individual first and second compounds;
  • the combination must have a solubility in 1% acetic acid at room temperature of at the most about 5% w/w such as, e.g., at the most about 4%, 3%, 2%, 1%, 0.5%, 0.25%, 0.1%, 0.05%, 0.01%, 0.005% or 0.001% w/w; and
  • the combination must have a solubility in water at room temperature of at the most about 5% w/w such as, e.g., at the most about 4%, 3%, 2%, 1%, 0.5%, 0.25%, 0.1%, 0.05%, 0.01%, 0.005% or 0.001% w/w.
  • tests may of course also be relevant such as, e.g., a test involving mass spectroscopy, or an in vitro model for testing the properties of the combination. Relevant in vitro models are described above. Such tests may either be employed with a view to determine any differences in the properties of a combination according to the invention compared with the properties of the individual compounds and a physical blend thereof, or they may be employed as in vitro models for the therapeutic activity/effect of a combination according to the invention.
  • the first compound is an oligo- or polysaccharide containing aminosugar units.
  • an aminosugar unit is a sugar unit wherein at least one of the hydroxy groups available in the sugar unit has been substituted by an amino group or an alkanoylated amino group such as an acetylated amino group.
  • the polysaccharide may contain unsubstituted sugar units or sugar units substituted with e.g. alkoxy (such as 2,3-dimethylglucose) or acyloxy.
  • the aminosugar units are alkanoylated. In some cases, it may also prove suitable that none of the aminosugar unit are alkanoylated.
  • the first compound preferably has a degree of animation in the range of 0.1-4, such as, e.g. in the range of 0.1-3 or 0.1-2, in particular in the range of 0.5-1.5 such as about 1, the degree of animation being defined as the average number of amino groups or alkanoylated amino groups per saccharide unit.
  • the first compound has a degree of animation of about 1.
  • a first compound is, e.g., a first compound wherein each sugar unit contains an amine or an alkanoylated amine group.
  • the individual sugar units of the first compound may be the same or different and are pentoses or hexoses.
  • pentoses include fructofuranose.
  • the hexoses are selected from the group consisting of glucose, mannose, galactose, xylose and ribose.
  • units like N- acetyl-D-glucosamine and/or N-acetylmuramic acid or the deacetylated compounds thereof may be incorporated in a first compound of the invention.
  • first compounds are compounds selected from the group consisting of chitosan, chitosans obtained by deacetylation of chitin to various degrees of deacetylation, chitosan derivatives, glycosaminoglycans including chondroitin, chondroitin sulfate, hyaluronic acid, dermatan sulfate and keratan sulfate; aminated dextrans including DEAE-dextran; aminated starch, aminated glycogen, aminated cellulose, aminated pectin, heparin, and salts, complexes, derivatives and mixtures thereof.
  • chitosan and chitosan derivatives are promising candidates as a first compound.
  • Water-soluble chitosan has been described as an agent in the treatment of wounds; the chitosan is said to prevent the formation of fibrin strands (US Patent No. 4,532,134).
  • Such a prevention of the formation of fibrin strands is believed to promote the proliferation of fibroblasts and the synthesis of collagen thereby allowing the promotion of normal tissue regeneration.
  • Chitosan has also been applied in the development of delivery systems as a means for obtaining controlled release of drugs (see e.g. WO 96/05810). Furthermore, chitosans have been found to have mucoadhesive properties (see e.g. EP 514 008, C-M. Lehr et al., "In vitro evaluation of mucoadhesive properties of chitosan and some other natural polymers, from C-M. Lehr's Ph.D. thesis, 1991, Leiden University, the Netherlands) and the use of chitosan as an enhancer has been suggested.
  • Chitosans are considered as biocompatible macromolecules due to assumed low toxicity and biodegradability. They are degraded by lysozymes and related enzymes, such as N-acetyl-D- glucosaminidases. The lysozymic degradation of chitosan is believed to increase with the degree of N-acetylation (see e.g. Aiba, Int. J. Biol. Macromol., 14, 1992, 225-228) and this property may influence the release rate of the second compound in a combination according to the invention (see below). Accordingly, chitosan for therapeutic use must be regarded as a safe substance.
  • Chitosan is a linear 1,4-bound polysaccharide built up from ⁇ -glucosamine entities.
  • the chitosan is manufactured by N-deacetylation of chitin which is a naturally occurring polymer forming the shell of inter alia insects and crayfish.
  • Commercial chitin is recovered from crab and shrimp shell which are waste products from the fishing industry. It is possible to manufacture chitosans of varying degrees of N-acetylation by controlling the reaction conditions (e.g. controlling the alkaline treatment of chitin).
  • chitosan may be manufactured with different molecular weights (Antonsen et al., Carbohydrate Polymers, 22, 1993, 193-201).
  • chitosan can be obtained by controlling the degradation of a high molecular weight chitosan in e.g. 1 N hydrochloric acid or by subjecting the chitosan to a controlled enzymatic degradation.
  • alkali such as sodium hydroxide
  • N- deacetylation takes place and the acetamido groups are converted into amino groups to form chitosan.
  • the aminosugar in the first compound of a combination according to the invention is totally or partly alkanoylated with acetamido groups or even in some cases all parent acetamido groups have been deacetylated to amino groups.
  • the degree of acetylation as well as the molecular weight of a chitosan have some impact on the properties of the combination formed.
  • the in vivo release of the second compound from a combination is subjected to at least one mechanism involving the degradation of the chitosan by an enzymatic process e.g. involving enzymes such as, e.g., lysozyme and proteases (present in wounds/ulcers), hyaluronidases, chitonases, metalloproteinases, collagenases, elastases and/or a combination thereof.
  • the lysozymic degradation of chitosan increases with the degree of N-acetylation and, accordingly, the release rate of the second compound in the combination may increase with the degree of N-acetylation.
  • Another release mechanism operating either alone or in combination with the above-mentioned mechanism may be based on a change in ionic strength; the higher the ionic strength is in the environment the faster the release is expected to take place due to the theory that the combination may be held together inter alia by ionic bondings and/or by inclusion. Without being bound to any theory the above-mentioned two release mechanisms seem to be most important.
  • release mechanisms seem to support the indication i) that the molecular weight of the chitosan comprised in the combination has an impact on the release rate (i.e. mainly for the release mechanism involving enzymatic degradation), and ii) that the degree of deacetylation of the chitosan comprised in the combination may have an impact on the release rate (i.e. probably for the release mechanism involving a change in ionic strength of the release medium).
  • the molecular weight as well as the degree of deacetylation also are important determinants for i) the biodegradability of a chitosan, ii) the toxicity, iii) the degree of mucoadhesiveness, and iv) the enhancer effect.
  • the first and/or second compound or even the combination by itself may exert its/their effect in undissolved stage e.g. in the form of fragments of molecules extending out from the particle surface.
  • fragments of dissolved second compound such as, e.g., sucrose octasulfate
  • first compound such as, e.g., chitosan
  • second compound such as, e.g., chitosan and/or the combination between the first and the second compound such as, e.g., chitosan-sucrose octasulfate, respectively, may also be effective with respect to wound healing.
  • sucrose octasulfate (representative of a second compound) is capable of exerting its effect when at least some of the chitosan (representative of a first compound) is degraded to smaller fragments; an ion-exchange process may be involved in the release/availability of sucrose octasulfate.
  • the first compound is a chitosan, especially a chitosan which has a molecular weight in a range of about 3,000 to about 1,500,000 daltons.
  • the chitosan notably has a degree of deacetylation of at the most 100% such as at the most 99%, 95%, 90%, 85%, or 80%, or expressed in another way the chitosan has a degree of deacetylation in a range of about 10-90% such as about 20-85%, about 30-80%, about 40-75%, about 50-75%, about 60-85%, about 75-85%, or about 80-90%.
  • the concentration of chitosan in the combination is in a range of from about 5 to about 99.99% w/w such as, e.g. from about 10-80% w/w, about 20-75% w/w, about 30-70% w/w or about 40- 70% w/w based on the total combination.
  • the first compound of a combination according to the invention is a polysaccharide containing aminosugar units.
  • first compounds may also prove suitable for use in a combination according to the invention provided that such first compounds are capable of reacting with a second compound (see below for details) to form a combination which is not merely a physical blend of the two individual compounds (evidenced as described above), which is capable of releasing the second compound e.g. in wounded tissue, and which is biocompatible and biodegradable.
  • a proviso for such other kinds of suitable first compounds may be found in the group consisting of e.g. natural and synthetic polymers containing at least one amino group.
  • the second compound of the combination is a polysulfated mono-, di- or oligosaccharide.
  • the monosaccharide may be, e.g., fructofuranose, glucose, mannose, galactose or ribose.
  • Particularly promising second compounds are polysulfated disaccharides, such as compounds based on, e.g., sucrose, a sucrose derivative or a complex or salt thereof.
  • the saccharide is polysulfated or even persulfated, which means that at least two and preferably more (such as all) hydroxy groups are sulfated.
  • the polysulfated saccharide is sucrose
  • the sucrose is at least tetrasulfated, preferably the sucrose is sucrose octasulfate or a salt or a complex thereof including the sodium salt thereof.
  • the sulfated saccharide may be complexed with or form a salt with an alkali metal (e.g. sodium or potassium), an alkaline earth metal (e.g. magnesium or barium) or another metal (e.g. aluminum, etc.)
  • an alkali metal e.g. sodium or potassium
  • an alkaline earth metal e.g. magnesium or barium
  • another metal e.g. aluminum, etc.
  • sucrose octasulfate When sucrose octasulfate is employed as a second compound in the combination, the content thereof in the combination is in a range of 0.01-95% w/w such as, e.g., in a range of about 20- 90% w/w, 25-80% w/w, 30-70% w/w or 30-60% w/w based on the weight of the total combination.
  • the combination is formed between a chitosan as the first compound and a sucrose octasulfate as the second compound.
  • the chitosan may be chitosan polymer having a molecular weight in a range of from 10 kdalton to 1500 kdalton and a degree of deacetylation in a range of at the most 100%.
  • Chitosan is presently available in qualities having a mean molecular weight in a range of from about 10 kdalton to about 1000 kdalton and a degree of deacetylation in a range of from about 75% to about 85%, but chitosans of other mean molecular weights and having other degrees of deacetylation are obtainable by employment of methods well known to a person skilled in the art (cf. above).
  • chitosans examples include chitosan, Wella “low viscosity”, C, Wella “high viscosity”, C, Dr. Knapzyk, Daichitosan H, Daichitosan VH, SeaCure 240, SeaCure 210, Chitosan (Sigma), Polycarbophil/daichitosan VH blend, Protasan CL 210, Protasan G210, and Protasan G110.
  • chitosans having a mean molecular weight of about 70,000-500,000 such as about 250,000-500,000 and a degree of N-acetylation of about 10-50% such as about 20-40% may be interesting candidates as first compounds in a combination according to the invention.
  • the chitosan may be in the form of a chitosan base or in the form of a salt such as, e.g., a glutamate, a lactate, or a hydrochloride salt.
  • a salt such as, e.g., a glutamate, a lactate, or a hydrochloride salt.
  • Mixtures of chitosan base and one or more chitosan salts are within the scope of the present invention as well as mixtures of chitosans having different mean molecular weights and/or degrees of deacetylation.
  • An interesting example of a combination according to the invention is a combination wherein the first compound is a chitosan having a molecular weight in a range from about 5,000 to about 100,000 dalton and the second compound is a sucrose octasulfate.
  • the combination is an active substance for use in medical treatment.
  • the combination may be used in the treatment or prevention of any relevant conditions, especially in conditions involving tissue repair such as in the healing of wounds.
  • the combination may be used in the treatment or prevention of any condition or disease where a sucrose octasulfate has a beneficial effect.
  • a sucrose octasulfate includes sodium or potassium sucrose octasulfate or the aluminium complex of sucrose octasulfate (known as sucralfate) or a combination thereof.
  • sucralfate sucralfate
  • the combination may also be used for the treatment or prevention of inflammatory conditions or for any condition which respond positively to a treatment with sucrose octasulfate.
  • chitosan is a biocompatible and relatively slowly biodegradable compound and sucrose octasulfate is also biodegradable. A combination of these two compounds is therefore believed also to be biodegradable. As mentioned above, chitosan is degraded in vivo by enzymes, e.g.
  • Lysozyme is present in a number of body fluids and is released by leukocytes and other cells normally present in wounds and other tissues where tissue repair is ongoing.
  • a working theory for a combination of chitosan and sucrose octasulfate is that when the combination is applied to a wound or a tissue which is to be repaired, then the lysozyme and/or protease present in the wound or tissue will degrade the combination, thereby releasing the sucrose octasulfate from the combination.
  • a combination will have a prolonged effect and less frequent administration is required compared to administration of sucrose octasulfate alone.
  • wound dressings are normally changed every other day or at the most once daily which limits the frequency of possible administration to wounds. With respect to internal wounds arising from surgery, the frequency of administration is generally very limited and normally only one administration is possible.
  • the presently most important use of a combination according to the invention is the use as a wound healing agent, i.e. an agent which accelerate, stimulate or promote healing of dermal or mucosal wounds.
  • tissue repair agent is another important use.
  • wound used in the present context denotes any wound (see below for a classification of wounds) and at any particular stage in the healing process including the stage before any healing has initiated.
  • a composition according to the invention will typically stabilise and/or stimulate fibroblast growth factors (FGF), the formation of collagen or the healing of wounds in vitro when subjected to relevant test like the tests described herein.
  • FGF fibroblast growth factors
  • wound denotes a bodily injury caused by physical means, with disruption of the normal continuity of structures.
  • wounds are, e.g., contused wounds, incised wounds, penetrating wounds, perforating wounds, puncture wounds, subcutaneous wounds, etc.
  • ulcer normally denotes a local defect, or excavation, of the surface of an organ or tissue, which is produced by the sloughing of inflammatory necrotic tissue.
  • ulcers are, e.g., peptic ulcer, duodenal ulcer, gastric ulcer, gouty ulcer, diabetic ulcer, hypertensive ischemic ulcer, stasis ulcer, sublingual ulcer, submucous ulcer, symptomatic ulcer, trophic ulcer, tropical ulcer, veneral ulcer, etc.
  • peptic ulcer duodenal ulcer
  • gastric ulcer gastric ulcer
  • gouty ulcer diabetic ulcer
  • hypertensive ischemic ulcer stasis ulcer
  • sublingual ulcer submucous ulcer
  • symptomatic ulcer trophic ulcer
  • tropical ulcer veneral ulcer
  • wounds In general wounds may be classified as follows:
  • Burns and chemical injuries e.g. superficial burns (first degree), deep dermal burns (second degree), and full thickness (third degree)]
  • leg ulcers venous, ischaemic or traumatic
  • wounds may also be classified by appearance as follows:
  • necrotic wound covered with a hard, dry black necrotic layer (e.g. small and superficial wounds or extensive and deep wounds)
  • clean wounds with significant tissue loss granulating wounds such as, e.g., clean surgical wounds with significant tissue loss, chronic wounds with low to moderate exudate, chronic open wounds with moderate to high exudate, and chronic flask-shaped wounds
  • malodorous wounds e.g. infected pressure sores, fungating carcinomas, etc.
  • Wounds/ulcers may be of a chronic or acute character.
  • Vascular, diabic and decubitis wounds belong to the group of chronic wounds, whereas e.g. burns belong to the group of acute wounds.
  • wounds may be a result of a physical injury, but the presence of wounds/ulcers may also be an indication of an underlying disease or disturbance such as, e.g., diabetes, hypertension, etc. (see e.g. Harrison's Principle of Internal Medicine, Eleventh Edition, McGraw-Hill Book Company, 1987 edited by E. Braunwald et al.)
  • the presence of leg and ankle ulcers as well as pressure sores like decubitus may also indicate that the physical activity and/or nutritional status of an individual is sub-optimal.
  • debridement and wound cleansing are of particular importance. It is believed that the cleaning and/or debridement of wounds/ulcers are a prerequisite for the healing process and, furthermore, when wound healing agents are applied such agents have to exert their effect on fresh and vital tissue and not on dead tissue or contaminated tissue.
  • Debridement of necrotic tissue can be performed by at least four different methods: i) sharp debridement, ii) mechanical debridement, iii) enzymatic debridement, and iv) autolytic debridement.
  • Sharp debridement is the most rapid method and may be the most appropriate technique for removing areas of thick, adherent eschar and devitalized tissue in extensive ulcers. Sharp debridement is normally performed in operating room and sterile instruments are used. Furthermore, clean, dry dressing should be applied.
  • Mechanical debridement includes the use of wet-to-dry dressing at prescribed intervals, hydrotherapy, wound irrigation and dextranomers.
  • Enzymatic debridement is often used in long-term care facilities and in home care. The method is not suitable for infected ulcers which should be debrided more rapidly. Enzymatic debridement is accomplished by applying topical debriding agents to devitalized tissue on the wound's surface. Collagenase and proteases are examples of debriding agents. A clean moist dressing should be applied over the ulcer after enzyme application.
  • Autolytic debridement is accomplished by placing a synthetic dressing over the ulcer and allowing the eschar to self-digest through the action of enzymes normally present in the wound fluid. Autolytic debridement is contraindicated in infected ulcers.
  • a combination according to the invention or a composition containing the combination can be applied either directly on or into the wound or ulcer or it can be applied in the form of a dry or moist, clean dressing into which the combination has been incorporated.
  • the combination according to the invention may of course also be applied in connection with cleansing of the ulcer/wound. Ulcer wounds should be cleansed initially and at every dressing change.
  • the process of cleansing a wound involves selecting a wound-cleansing solution and a mechanical means of delivering that solution to the wound.
  • a combination according to the present invention may be used for any wound/ulcer independent of its ethiology, nature or healing stage. It may be used for curative purposes as well as for preventive purposes. Furthermore, a combination according to the invention may be used together with other active substances such as, e.g., antiviral, antiinflammatory and/or antibacterial substances, hormones, growth factors, enzymes, and mixtures thereof. Examples of active substances for use together with a combination of the present invention are:
  • antiviral substances acyclovir, famciclovir, deciclovir, penciclovir, zidovudin, ganciclovir, didanosin, zalcitabin, valaciclovir, sorivudine, lobucavir, brivudine, cidofovir, n-docosanol, ISIS- 2922, and prodrugs and analogues thereof, amantadin, rimantadin, foskarnet, idoxuridin, fluoruracil, interferons and variants thereof, including alpha interferon, beta interferon, and gamma interferon, tromantadin, lentinan, levofloxacin, stavudine, tacrine, vesnarinone, ampligen, atevirdine, delavirdine, hydroxyurea, indinavir sulfate, interleukin-2 fusion toxin, seragen, lamivudine, lid
  • antifungal and/or antiprotozoal substances such as, e.g., miconazol, ketoconazole, clotrimazole, amphotericin B, nystatin, mepyramin, econazol, fluconazol, flucytocine, griseofulvin, bifonazole, amorolfine, mycostatin, itraconazole, terbenafine, terconazole, tolnaftate, fucidin, erythromycin, macrolides, metronidazole, and the like;
  • NSAID's such as, e.g., ibuprofen, indomethacin, naproxen, diclofenac, tolfenamic acid, piroxicam, and the like;
  • antibiotic such as, e.g., amikacin, amikacin sulfate, amoxycillin, amphomycin, ampicillin, avoparcin, azidocillin, bacampicillin, bacitracin, bakanamycin, benzanthine penicillin, benzanthine phenoxymethylpenicillin, benzylpenicillin, BL-P 1654, carbenicillin sodium, carfecillin sodium, carindacillin sodium, cefaclor, cefadroxil, cefapirin sodium, cefatrizine, cefazaflur sodium, cefoperazone sodium, cefotaxime sodium, cefoxitin sodium, cephacetrile sodium, cephalexin, cephaloglycin, cephalonium, cephaloridine, cephalothin sodium, cephamandole, cephazolin sodium, chloramphenicol, chlortetracycline, ciclacillin, clavulanic acid, clemizole penicillin,
  • hormonal substances such as, e.g., estradiol, estron, estriol, polyestradiol, polyestriol, dienestrol, diethylstilbestrol, progesterone, dihydroergosterone, cyproterone, danazol, testosterone, and the like;
  • corticosteroids such as, e.g., beclomethasone, betamethasone, betamethasone-17-valerate, betamethasone-dipropionate, clobetasol, clobetasol-17-butyrate, clobetasol-propionate, desonide, desoxymethasone, dexamethasone, diflucortolone, flumethasone, flumethasone-pivalate, fluocinolone acetonide, fluocinonide, hydrocortisone, hydrocortisone- 17-butyrate, hydrocortisone- buteprate, methylprednisolone, triamcinolone acetonide, budesonide, halcinonide, fluprednide acetate, alklometasone-dipropionate, fluocortolone, fluticason-propionate, mometasone-furate, desoxymethasone, diflurason-
  • disinfectants like e.g. chlorhexidine, hydrogen peroxide, proflavine, cetremide, benzylperoxide, potassium permanganate, iodine, iodine derivatives, silver sulfadiazine, and hypochlorite;
  • growth factors enzymes and/or peptides such as, e.g., growth hormone releasing factors, growth factors (epidermal growth factor (EGF), nerve growth factor (NGF), TGF, PDGF, insulin growth factor (IGF), fibroblast growth factors ( ⁇ FGF, ⁇ FGF, etc.), and the like), somatostatin, calcitonin, insulin, vasopressin, interfeons, IL-2, urokinase, serratiopeptidase, streptokinase, superoxide dismutase (SOD), thyrotropin releasing hormone (TRH), luteinizing hormone releasing hormone (LH-RH), corticotrophin releasing hormone (CRF), growth hormone releasing hormone (GHRH), oxytocin, erythropoietin (EPO), colony stimulating factor (CSF), and the like.
  • growth hormone releasing factors growth factors (epidermal growth factor (EGF), nerve growth factor (NG
  • a composition according to the invention may contain a free amount of first and/or second compound.
  • a free amount of a wound healing agent such as, e.g. sucrose octasulfate and/or chitosan may in certain cases be desired especially in those cases where a rapid onset of the effect is necessary or desirable or, alternatively, in those cases where a burst effect is desired.
  • the combination is applied together with at least one other active substance (as mentioned above) the dose of the other active substance is in a range necessary to obtain a therapeutic effect.
  • the dose required is the same as in pharmaceutical compositions where no combination according to the invention is present.
  • the dose necessary to obtain a suitable response can be determined in an analogous manner as described below for the determination of the dose of the combination.
  • compositions comprising a combination according to the invention together with an active drug substance which is effective against any condition which may be the underlying cause of the ulcer.
  • Such a composition would have two objectives, namely i) to impart wound healing and ii) to treat the underlying disease to the formation of ulcers.
  • the wound is used as the application site for delivery of a further active drug substance to the circulatory system. It is contemplated that the access to the circulatory system is much easier via wounds/ulcers than via intact skin.
  • Vaccines are also interesting candidates for a delivery to the circulatory system via a wound/ulcer. Diseases like malignant melanom and psoriasis are examples of diseases which may be treated by delivery of the active drug substance via an ulcer.
  • compositions designed for gene therapy are topical compositions designed for gene therapy (local or systemic).
  • a combination according to the invention is preferably formulated in a pharmaceutical composition containing the composition and, optionally, one or more pharmaceutically acceptable excipients.
  • the invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a combination according to the invention.
  • a composition may also contain one or more substances having any one of the activities mentioned above or any other suitable activity.
  • compositions may be in form of, e.g., a spray, a solution, a dispersion, a suspension, an emulsion, tablets, capsules, pills, powders, granulates, gels including hydrogels, lotions, pastes, ointments, creams, drenches, dressings, hydrogel dressings, hydrocolloid dressings, films, foams, sheets, bandages, plasters, delivery devices, suppositories, enemas, implants, aerosols, microcapsules, microspheres, nanoparticles, liposomes, and in other suitable form.
  • Compositions for application to the skin or to the mucosa are considered most important in connection with the present invention.
  • composition comprising the combination to be administered may be adapted for administration by any suitable route, for example by topical (dermal), oral, buccal, nasal, aural, rectal, vaginal, pulmonal administration, or by administration to a body cavity such as, e.g., a tooth root.
  • a composition may be adapted to administration in connection with surgery, e.g. in connection with incision within the body in order to promote healing of internal wounds and tissue damage such as bone fractures.
  • compositions may be formulated according to conventional pharmaceutical practice, see, e.g., "Remington's Pharmaceutical Sciences” and “Encyclopedia of Pharmaceutical Technology", edited by Swarbrick, J. & J. C. Boylan, Marcel Dekker, Inc., New York, 1988.
  • a composition comprising a combination according to the invention is intended for skin or mucosa.
  • Other applications may of course also be relevant such as, e.g., application on dentures, protheses and application to body cavities such as the oral cavity.
  • the mucosa is preferably selected from oral, nasal, aural, lung, rectal, vaginal, and gastrointestinal mucosa.
  • application within the dental/odontologic area is also of great importance. Relevant examples are application to periodontal (dental) pockets, to gigiva or to gigival wounds or ulcers, or in connection with dental surgery.
  • the pharmaceutical composition comprising a combination according to the invention serves as a drug delivery system.
  • drug delivery system denotes a pharmaceutical composition (a pharmaceutical formulation or a dosage form) which upon administration presents the active substance to the body of a human or an animal.
  • drug delivery system embraces plain pharmaceutical compositions such as, e.g., creams, ointments, liquids, powders, tablets, etc. as well as more sophisticated formulations such as sprays, plasters, bandages, dressings, devices, etc.
  • compositions according to the invention may comprise pharmaceutically or cosmetically acceptable excipients.
  • the pharmaceutically acceptable excipient for use in a composition is generally dependent on the dosage form suitable for use for a particular kind of wound/ulcer. In the following is given a detailed list of suitable pharmaceutically acceptable excipients for use in compositions according to the invention.
  • a pharmaceutical compositions according to the invention may be adapted to administration via the oral, buccal, mucosal, nasal, rectal, vaginal, or topical route or to application directly on wounds and/or ulcers.
  • the formulations according to the invention may contain conventionally non-toxic pharmaceutically acceptable carriers and excipients including microspheres and liposomes.
  • the compositions include creams, ointments, hydrophilic ointments, lotions, liniments, gels, hydrogels, solutions, suspensions, sticks, sprays, pastes, plasters, films, powders, soaps, shampoos, jellies, dressings such as absorbent wound dressings, pads, bandages, foams, plasters, and transdermal drug delivery systems.
  • the pharmaceutically acceptable excipients may include emulsifying agents, antioxidants, buffering agents, preservatives, humectants, penetration enhancers, chelating agents, gelforming agents, ointment bases, perfumes, and skin protective agents.
  • emulsifying agents are naturally occurring gums, e.g. gum acacia or gum tragacanth; naturally occurring phosphatides, e.g. soybean lecithin; sorbitan monooleate derivatives; wool fats; wool alcohols; sorbitan esters; monoglycerides; and fatty alcohols.
  • antioxidants examples include butylated hydroxy anisole (BHA), ascorbic acid and derivatives thereof, tocopherol and derivatives thereof, butylated hydroxy anisole, and cysteine.
  • Suitable examples of preservatives for use in compositions according to the invention are parabens, such as methyl, ethyl, propyl p-hydroxybenzoate, butylparaben, isobutylparaben, isopropylparaben, potassium sorbate, sorbic acid, benzoic acid, methyl benzoate, phenoxyethanol, bronopol, bronidox, MDM hydantoin, iodopropynyl butylcarbamate, EDTA, propyleneglycol (increases the solubility of preservatives) benzalconium chloride, and benzylalcohol, or mixtures of preservatives like Germaben II & 1TE (mixture of imidazolidinyl urea, methyl and propyl parabens and propylene glycol; available from ISP-Sutton Labs.)
  • humectants examples include glycerin, propylene glycol, sorbitol, lactic acid, and urea.
  • chelating agents examples include sodium EDTA, citric acid, and phosphoric acid.
  • excipients are edible oils like almond oil, castor oil, cacao butter, coconut oil, corn oil, cottonseed oil, linseed oil, olive oil, palm oil, peanut oil, poppyseed oil, rapeseed oil, sesame oil, soybean oil, sunflower oil, and teaseed oil; and of polymers such as carmelose, sodium carmelose, hydroxypropylmethylcellulose, hydroxyethylcellylose, hydroxypropylcellulose, pectin, xanthan gum, carrageenan, locust bean gum, acacia gum, gelatin, carbomer, emulsifiers like vitamin E, TPGS, glyceryl stearates, cetanyl glucoside, and alginates.
  • edible oils like almond oil, castor oil, cacao butter, coconut oil, corn oil, cottonseed oil, linseed oil, olive oil, palm oil, peanut oil, poppyseed oil, rapeseed oil, sesame oil, soybean oil, sunflower oil,
  • ointment bases in general, are beeswax, paraffin, cetanol, cetyl palmitate, vegetable oils, sorbitan esters of fatty acids (Span), polyethylene glycols, and condensation products between sorbitan esters of fatty acids and ethylene oxide, e.g. polyoxyethylene sorbitan monooleate (Tween).
  • hydrophobic or water-emulsifying ointment bases are paraffins, vegetable oils, animal fats, synthetic glycerides, waxes, lanolin, and liquid polyalkylsiloxanes.
  • hydrophilic ointment bases are solid macrogols (polyethylene glycols).
  • ointment bases are triethanolamine soaps, sulphated fatty alcohol and polysorbates.
  • gel bases or components which are able to take up exudate from a wound/ulcer are: liquid paraffin, polyethylene, fatty oils, colloidal silica or aluminium, zinc soaps, glycerol, propylene glycol, tragacanth, starch, cellulose derivatives, carboxyvinyl polymers, magnesium- aluminium silicates, Carbopol, hydrophilic polymers such as, e.g. starch or cellulose derivatives, liquid absorbing wound bandages, water-swellable hydrocolloid, and alginates.
  • Examples of powder components are: alginate, collagen, lactose, powder which is able to form a gel when applied to a wound/ulcer (absorbs liquid/wound exudate). Normally, powders intended for application on large open wounds must be sterile and the particles present must be micronized.
  • Alginic acid or alginates are important excipients in connection with pharmaceutical compositions according to the present invention. As explained above, alginates can form a gel and, furthermore, they can absorb wound exudates and thus contributing to controlling the moisture content of a wound. Alginic acid and alginates are available in various qualities having a mean molecular weight in the range of e.g. about 32,000-200,000. The relative proportion of mannuronic and guluronic acid residues varies from one quality to another.
  • excipients for use in topical compositions include tackifier resin, viscous elastomeric binders, elastic film, elastic adhesive material, elastomers and plasticizers.
  • Dressings and/or bandages are also important delivery systems for a combination according to the invention.
  • Dressings may be in the form of absorbent wound dressings for application to exuding wounds. Such dressings are frequently made of cotton or viscose fibres which are enclosed in a sleeve of gauze or a suitable non-wowen fabric. Other relevant materials are cellulose fibres, cellulose wood pulp (fine powdery material).
  • Dressings may also be in the form of hydrogel dressings such as, e.g., i) dressings having a fixed three-dimensional macro- structure, and ii) dressings involving amorphous hydrogels.
  • the dressings progressively decrease in viscosity when the material absorbs fluid, and the dressing may then flow on top of the wound and take on the shape of the wound.
  • examples of other kinds of dressings are i) hydrocolloid dressings (gel-forming agents combined with other material such as, e.g.
  • elastomers and adhesives including hydrocolloid granules or paste and hydrocolloid sheet, ii) alginate sheet (rope or ribbon, alginate with integral absorbent pad), iii) foams (foam dressings, silastic foam, polyurethane foam), iv) various polysaccharide materials, v) occlusive dressings, vi) semipermeable dressings, vii) paraffin gauze dressings, viii) Tulle dressings, ix) polysaccharide pastes, granules and beads (may be manufactured from dextran derivatives), and x) odour-absorbing dressings.
  • hydrocolloid granules or paste and hydrocolloid sheet ii) alginate sheet (rope or ribbon, alginate with integral absorbent pad), iii) foams (foam dressings, silastic foam, polyurethane foam), iv) various polysaccharide materials, v) occlusive dressings, vi
  • Suitable bandages may be i) non-extensible bandages, ii) extensible bandages, iii) adhesive/cohesive bandages, iv) tubular bandages, v) medicated paste bandages, and vi) orthopaedic casting materials.
  • Semipermeable films and thin foam sheets have little or no intrinsic absorbent capacity. They are permeable to moisture vapour when placed on a wound and, accordingly, the aqueous component of a wound exudate is lost through the backing of the dressing in the form of a vapour. The cellular material remains trapped at the surface of the wound.
  • Alginate and hydrocolloid dressings take up wound exudate when placed on a wound. When doing so they produce an aqueous gel on the surface of the wound and this gel is believed to be beneficial for the healing of the wound due to the retaining of moisture in the wound.
  • the compositions mentioned above for topical administration are most suitably for application directly to wounds or they may be suitable for application to or for introduction into relevant orifice(s) of the body, e.g. the rectal, urethral, vaginal aural, nasal or oral orifices.
  • the composition may simply be applied directly on the part to be treated such as, e.g., on the mucosa, or by any convenient route of administration.
  • Fluid/liquid compositions for oral use or for application to mucosa or skin are provided.
  • compositions like a suspension, an emulsion or a dispersion provide the combination in admixture with a dispersing or wetting agent, suspending agent, and/or one or more preservatives.
  • Such compositions may also be suitable for use in the delivery of the combination to e.g. an intact or damaged mucosa such as the gastrointestinal, buccal, nasal, rectal, or vaginal mucosa, or for administration to intact or damaged skin, or wounds/ulcers.
  • Suitable dispersing or wetting agents are, for example, naturally occurring phosphatides, e.g., lecithin, or soybean lecithin; condensation products of ethylene oxide with e.g. a fatty acid, a long chain aliphatic alcohol, or a partial ester derived from fatty acids and a hexitol or a hexitol anhydride, for example polyoxyethylene stearate, polyoxyethylene sorbitol monooleate, polyoxyethylene sorbitan monooleate, etc.
  • naturally occurring phosphatides e.g., lecithin, or soybean lecithin
  • condensation products of ethylene oxide with e.g. a fatty acid, a long chain aliphatic alcohol, or a partial ester derived from fatty acids and a hexitol or a hexitol anhydride for example polyoxyethylene stearate, polyoxyethylene sorbitol monooleate, polyoxyethylene sorbitan
  • Suitable suspending agents are, e.g., naturally occurring gums such as, e.g., gum acacia, xanthan gum, or gum tragacanth; celluloses such as, e.g., sodium carboxymethylcellulose, microcrystalline cellulose (e.g. Avicel® RC 591, methylcellulose; alginates such as, e.g., sodium alginate, etc.
  • naturally occurring gums such as, e.g., gum acacia, xanthan gum, or gum tragacanth
  • celluloses such as, e.g., sodium carboxymethylcellulose, microcrystalline cellulose (e.g. Avicel® RC 591, methylcellulose
  • alginates such as, e.g., sodium alginate, etc.
  • compositions according to the invention are the same as those mentioned above.
  • compositions according to the invention include suppositories (emulsion or suspension type), enemas, and rectal gelatin capsules (solutions or suspensions).
  • suppository bases include cocoa butter, esterified fatty acids, glycerinated gelatin, and various water-soluble or dispersible bases like polyethylene glycols and polyoxyethylene sorbitan fatty acid esters.
  • additives like, e.g., enhancers or surfactants may be incorporated.
  • Nasal compositions For application to the nasal mucosa, nasal sprays and aerosols for inhalation are suitable compositions according to the invention.
  • the combination is present in the form of a particulate formulation optionally dispersed in a suitable vehicle.
  • the pharmaceutically acceptable vehicles and excipients and optionally other pharmaceutically acceptable materials present in the composition such as diluents, enhancers, flavouring agents, preservatives, etc. are all selected in accordance with conventional pharmaceutical practice in a manner understood by the persons skilled in the art of formulating pharmaceuticals.
  • the active substance may be adsorbed on the nasal mucosa.
  • the adsorption on the mucosa is believed to lead to a less irri- tative effect than when, e.g., a liquid vehicle, e.g. containing a penetration enhancer or promoter, is employed.
  • compositions for oral use are provided.
  • compositions according to the invention are may also be administered by the oral route.
  • Suitable oral compositions may be in the form of a particulate formulation or in the form of a solid, semi-solid or liquid dosage form.
  • compositions for oral use include solid dosage forms such as, e.g., powders, granules, sachets, tablets, capsules, effervescent tablets, chewable tablets, lozenges, immediate release tablets, and modified release tablets as well as fluid or liquid formulations such as, e.g. powders, dispersible powders, or granules suitable for preparation of an aqueous suspension by addition of an aqueous medium, emulsions, dispersions, and mixtures.
  • solid dosage forms such as, e.g., powders, granules, sachets, tablets, capsules, effervescent tablets, chewable tablets, lozenges, immediate release tablets, and modified release tablets as well as fluid or liquid formulations such as, e.g. powders, dispersible powders, or granules suitable for preparation of an aqueous suspension by addition of an aqueous medium, emulsions, dispersions, and mixtures.
  • compositions contain the combination and any further active substance optionally in admixture with one or more pharmaceutically acceptable excipient.
  • excipients may be, for example,
  • inert diluents or fillers such as sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starches including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate;
  • granulating and disintegrating agents for example, cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates, or alginic acid; binding agents, for example, sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, carboxymethylcellulose sodium, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone such as, e.g, PVP K12, PVP K15, PVP K17, PVP K25, PVP K30, PVP K60, PVP K90, or PVP K120, or combinations thereof, polyvinylacetate, or polyethylene glycol; and
  • lubricating agents including glidants and antiadhesives, for example, magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenated vegetable oils, or talc.
  • compositions can be colorants, flavouring agents, plasticizers, humectants, buffering agents, etc.
  • the unit dosage form may be provided with a coating like one or more of the coatings mentioned below.
  • the composition or the individual units or a tablet or a capsule containing the individual units may be coated e.g. with a sugar coating, a film coating (e.g. based on hydroxypropyl methylcellulose, methylcellulose, methyl hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, acrylate copolymers (Eudragit), polyethylene glycols and/or polyvinylpyrrolidone) or an enteric coating (e.g.
  • a time delay material such as, e.g., glyceryl monostearate or glyceryl distearate may be employed.
  • a combination is generally present in a concentration in a range of from about 0.01% to about 99.9% w/w.
  • the amount of composition applied will normally result in an amount of the second compound per cm 2 wound/skin/tissue area corresponding to from about 0.1 mg/cm 2 to about 500 mg/cm 2 such as from about 1 mg/cm 2 to about 100 mg/cm 2 .
  • the amount of composition applied will normally equals an amount of sucrose octasulfate per cm 2 wound/skin/tissue area corresponding to from about 0.1 mg/cm 2 to about 100 mg/cm 2 , such as from about 0.1 mg/cm 2 to about 10 mg/cm 2 .
  • the amount applied of the composition depends on the load and the release rate of e.g. the sucrose octasulfate on the first compound (e.g. chitosan), but is generally in a range corresponding to at the most 5 mg/cm 2 .
  • the concentration of the combination in a pharmaceutical composition depends on the nature of the second compound in question, its potency, the severity of the disease to be prevented or treated, and the age and condition of the patient. Methods applicable to selecting relevant concentrations of the combination in the pharmaceutical composition are well known to a person skilled in the art and may be performed according to established guidelines for good clinical practice (GCP) or Investigational New Drug Exemption ("IND") regulations as described in e.g. Drug Applications, Nordic Guidelines, NLN Publication No. 12, Nordic Council on Medicines, Uppsala 1983 and Clinical Trials of Drugs, Nordic Guidelines, NLN Publication No. 11, Nordic Council on Medicines, Uppsala 1983.
  • GCP clinical practice
  • IND Investigational New Drug Exemption
  • the invention in another aspect, relates to a method for the preparation of a combination, the method comprising mixing the first and the second compound in a suitable medium such as an aqueous medium and, optionally, adjusting the pH in order to precipitate the combination.
  • a suitable medium such as an aqueous medium
  • the precipitate is collected and, optionally, purified.
  • Other methods e.g., involving the preparation of a film, are apparent from the experimental section. It is contemplated that the loading of the second compound in the resulting combination as well as the dominating structure of the combination is dependent on the conditions prevailing during the reaction. Furthermore and as will appear from the experimental section herein, the reaction conditions are also believed to influence the composition and the yield of the combination obtained such as, e.g., the method of preparation (e.g.
  • the mixing process, the mixing process and the order in which the components is added the ionic strength prevailing in the reaction mixture, the viscosity, the kind of solvent chosen for the reactants, any addition of accelerators, the ratio of SOS and chitosan, the pH prevailing in the reaction mixture, reaction time, rotation speed, e.g. from 0-1000 rpm, homogenisation of the reaction mixture, heating/cooling of the reaction mixture, employment of ultrasonic treatment (cf.
  • Example 5 the starting materials (chitosan, chitosan glutamate, chitosan chloride, chitosan lactate, sucrose octasulfate as hydrate or other solvates or as a sodium, potassium or another salt, or derivatives thereof) and the concentration thereof, the precipitation method, any purification step, etc.)
  • the second compound is an active drug substance.
  • the invention relates also to a combination of a first compound of an oligo- or polysaccharide containing aminosugar units and a second compound of an active drug substance.
  • a combination is also not merely a physical blend but a reaction has taken place between the first and the second compound.
  • the combination may as well be a salt, a complex such as an inclusion complex, or the compounds may be held together in a polymeric network or as a mechanical inclusion, or the compounds may even be covalently bound together in the combination, or a combination of the above-mentioned mechanisms may be involved.
  • a combination as defined above must fulfil the same requirements as described above under the aspect of the invention relating to combinations wherein the first compound is as defined above but the second compound is a sulfated mono-, di- or oligosaccharide.
  • the second compound of a combination according to the invention is an active drug substance administration of the combination to a mammal, in particular a human, will result in a controlled release of the active drug substance due to a controlled release of the active substance from the combination (as discussed herein).
  • the combination may be bioadhesive due to the properties of the first compound (which e.g. may be a chitosan as described above). Administration of a bioadhesive combination may enable a localized effect as the combination is maintained at the application site by means of bioadhesion and from that site releases the active drug substance in a controlled manner.
  • active drug substances are able to form a combination as defined above when reacted with a first compound as defined above.
  • a characteristic of the active drug substances relevant in the present context as second compounds is that the active drug substances contain a carboxylic acid or a sulfonic acid group.
  • NSAIDs Non-Steroidal .Anti-inflammatory Drugs
  • Antihypertensive agents Alacapril Candesartan Carmoxirole Cilazapril Delapril Eprosartan Fosinopril Lisinopril Moveltipril Perindopril Ramipril Valsartan
  • Thyroid inhibitors 3, 5-diiodotyrosine Tiratricol (antihypothyroid)
  • Antineoplastic agents Chlorambucil
  • Epalrestat Epalrestat
  • Antihistamines Bietanantine Cetirizine Fexofenadine Acrivastine Tranilast
  • Vitamins Folic acid Pantothenic acid Vitamin A acid
  • CNS stimulants Amineptine Hexacyclonate sodium
  • ACE inhibitors Captopril Benazepril Enalapril Perindopril Trandolapril Moexipril Fosinopril Ramipril Lisinopril
  • Antidiabetic agents Repaglinide Tolrestat Zopolrestat
  • Especially interesting combinations according to the invention are combinations wherein the first compound is chitosan and the second compound is any of the above-mentioned active drug substances or combinations thereof.
  • combinations according to the invention wherein the second compound is an active drug substance as defined above are contemplated to act as prodrugs of the parent active drug substance and/or a synergistic effect between the first and the second compound may be achieved.
  • This means that combinations wherein the second compound is an active drug substance as defined above are suitable for the same therapeutic or prophylactic purpose as the parent drug substance.
  • the active drug substance may also be effective while being maintained in the combination; thus, e.g. fragments of the active drug substance may extend out from the combination and lead to a therapeutic effect. Accordingly, this aspect of the invention is not restricted to combinations which are suitable for use as wound healing agents but for any appropriate therapeutic/prophylactic use.
  • the combination according to the invention acts as a prodrug of the parent active drug substance
  • a person skilled in the art will have no difficulties in determining the dose of the combination to be administered to a mammal, in particular to a human, and in what form the dosage should be presented (e.g. compositions suitable for oral, nasal, topical, parenteral etc. administration).
  • the invention also relates to a method for the preparation of the abovementioned combinations wherein the second compound is an active drug substance as defined above and to pharmaceutical compositions containing the combinations together with at least one acceptable pharmaceutical excipient.
  • first aspect of the invention i.e. a combination between a first and a second compound, wherein the first compound is an oligo- or polysaccharide containing aminosugar units and the second compound is a sulfated mono-, di- or oligosaccharide
  • first compound is an oligo- or polysaccharide containing aminosugar units
  • second compound is a sulfated mono-, di- or oligosaccharide
  • Fig. 1 shows a thermogram of sodium sucrose octasulfate.
  • the thermogram was recorded on a Perkin Elmer Analysis 7 Series System using 6.58 mg of sodium sucrose octasulfate.
  • the start temperature was 20°C and the end temperature 200°C.
  • the plot shows the heat flow in mW against the temperature (°C).
  • Fig. 2 shows a thermogram of chitosan.
  • the experimental details are as described under Fig. 1. 1.600 mg of chitosan was used.
  • Fig. 3 shows a thermogram of a combination of chitosan and SOS (batch No. AAAS-2-1). The experimental details are as described under Fig. 1. 5.640 mg of chitosan-SOS was used.
  • Fig. 4 shows a thermogram of a physical blend of chitosan and sodium sucrose octasulfate. The experimental details are as described under Fig. 1. The physical blend used consisted of about 4 mg chitosan and about 21 mg sodium sucrose octasulfate.
  • Fig. 5 shows the results of a number of syntheses of chitosan-SOS in which the initial concentration of sodium sucrose octasulfate has been varied in order to determine whether different loads of SOS are obtained.
  • the graph is a plot of the loading of sodium sucrose octasulfate (Na-SOS) to chitosan (solution of Protasan Cl 210 2 mg/ml) versus the initial concentration of sodium sucrose octasulfate in the SOS solution used in the reaction mixture.
  • the different grades of chitosan and chitosan salts are specified by the mean molecular weight, the degree of deacetylation and viscosity of the solutions.
  • the viscosities given above are data from the supplier (1% solution in 1% w/v acetic acid, Brookfield LVT viscometer, 25°C, 30 rpm)).
  • Pronova Biopolymer a.s., Norway supplies two chitosan salts, namely the glutamate and the hydrochloride salt, both in two different qualities.
  • Protasan is a very purified product and Seacure is a product of regular quality.
  • the degree of deacetylation of chitosan and chitosan salts is normally about 84%
  • Methyl cellulose (Methocel MC) from Fluka (available form Bie & Berntsen, Denmark)
  • Coulter Multisizer II (Coulter), Malvern 2600 droplet and particle size analyse (for the determination of particle size distribution).
  • Str ⁇ lein Areameter and Coulter S3100 for the determination of the surface area of the particles.
  • the dynamic viscosity of a test sample or a composition is determined using a RheoStress RS 100 Rheometer, HAAKE (Germany) equipped with a RS 100 1.2 software package.
  • the measurement can be performed at 20°C** ⁇ 0.5°C or, alternatively, at 25°C or at 37°C ⁇ 0.5°C or at varying temperatures in order to obtain temperature curves.
  • Chitosan base is not soluble at pH above 7.0 and, therefore, the pH must be maintained at pH 6.0 or below to avoid precipitation.
  • a 1% w/v solution is prepared by slurring e.g. 2 g chitosan (Sigma, practical grade) in 100 ml of distilled water followed by adding 100 ml of a 2% w/v solution of acetic acid.
  • the resulting mixture i.e. a 1% w/v solution of chitosan in 1% w/v acetic acid, is stirred vigorously for 60 minutes or until dissolution has taken place. Heating the solution can accelerate the dissolution process, but prolonged heating may result in a decrease in viscosity.
  • chitosan can be dissolved directly by adding chitosan to a 1% w/v acetic acid solution.
  • 2% w/v solutions of chitosan (Sigma, practical grade) in 1% w/v can also be prepared.
  • chitosan may be dissolved under vigorous stirring for 24-48 hours.
  • a solution of 1% w/v (10 mg/ml) chitosan (Sigma) in 1% w/v acetic acid is pumped dropwise through a tip from a microliter pipette (Finntip 60) to a solution of 12.5 mg/ml sucrose octasulfate sodium salt (SOS-Na 8 ) in distilled water.
  • Beads (drops or pearls) appear in the solution as chitosan gets into contact with SOS-Na 8 presumably forming a combination or complex on the surface of the chitosan beads.
  • the beads are approximately 2 mm in diameter and the surface becomes opaque whitish (dependent on the concentration of SOS-Na 8 in the solution). Upon resting in the solution the beads lose their surface tension and shrink. The beads are then filtered and washed with distilled water followed by drying at 40°C in an oven. The dry product appears as shrivelled beads. It is a very hard material which is not easily pulverized.
  • sucrose octasulfate sodium salt SOS-Na 8
  • SOS-Na 8 sucrose octasulfate sodium salt
  • chitosan employed may be either chitosan base, chitosan hydrochloride or chitosan glutamate. In the following chitosan has been employed as the base unless otherwise specified.
  • Method I 10 ml of I (se above) is added slowly and dropwise to 40 ml of II at room temperature. The 10 ml of I is added over a time period of 40 min. The reaction time is 40 min. Small transparent spheres containing the chitosan solution are formed. The chitosan solution reacts slowly with sucrose octasulfate on the surface of the drops and small spheres are formed and filtered of. The thus collected spheres are dried at room temperature for 24 hours and dried flakes are obtained.
  • Method II 40 ml of II (5 mg/ml) is added to 10 ml of I in a continuous thin jet through a tip for a microliter pipette (Finntip 60) with continuously stirring with a Heidolph R2R-2000 stirrer (200-750 rpm). The formed combination (concentrated around the rotor-blades) is isolated and dried at room temperature for 24 hours.
  • Method III 10 ml of I is placed in a petri dish. The solution is dried 4 hrs in a fume cupboard or for 2 hours at 60°C. The chitosan solution is dried in an oven (40-60°C) until nearly dryness and a soft film is obtained. Alternatively, the chitosan solution is dried to dryness either at room temperature or in an oven (conditions as above) and if a crisp film is obtained, the film can easily be softened with water. To the thus obtained chitosan film, 20 ml of II (12.5 mg/ml) is added and left to evaporate the liquid,
  • AAAS5-1 0.5 g SOS-Na 8 dissolved in 100 ml of distilled water is added to 50 ml of a chitosan solution (cf. Example 1) by means of a syringe and a canula and with vigorously stirring. The precipitate is filtered off and dried at ambient temperature.
  • AAAS5-2 As AAAS5-1 but the chitosan solution is added to the SOS-Na s -solution.
  • AAAS7-2 10 ml of a chitosan solution (cf. Example 1) is dropwise added to 100 mg SOS-Na 8 in 40 ml distilled water.
  • AAAS7-6 10 ml of a chitosan solution (cf. Example 1) is dropwise added to 0.5 g SOS-Na 8 in 40 ml distilled water.
  • AAASll-3 0.100 g SOS-Na 8 in 40 ml distilled water is slowly added through a tip for a pipette to 10 ml of a chitosan solution (cf. Example 1), yield 0.1280 g.
  • AAASll-6 0.500 g SOS-Na 8 in 40 ml distilled water is slowly added through a tip for a pipette to 10 ml of a chitosan solution (cf. Example 1), yield 0.1749 g.
  • AAAS12-4 10 ml of a chitosan solution (cf. Example 1) is dropwise added to 0.150 g SOS-Na 8 in 40 ml distilled water, yield 0.1825 g.
  • AAAS12-6 10 ml of a chitosan solution (cf. Example 1) is dropwise added to 0.500 g SOS-Na 8 in 40 ml distilled water, yield 0.1638 g.
  • AAAS13b-4 a chitosan film is formed by completely drying of a 1% chitosan solution. Then 0.4 g SOS-Na 8 dissolved in 20 ml water is added.
  • AAAS13C-3 a film formed by SOS-Na 8 prepared by Method III given above in the reverse order.
  • AAAS15-1 and AAAS15-3 A chitosan solution (cf. Example 1) is poured into a petri dish and a chitosan film is obtained after evaporation of the solvent. Then a solution of SOS-Na 8 is added (i.e. equivalent to Method III given above)
  • chitosan has been employed in form of its glutamate or its hydrochloride.
  • the initial results indicate that the viscosity of the reaction solution obtained by employment of these salts is too low to obtain a spheric product. Employment of the salts may therefore involve use of a viscosity adjusting agent if a spheric product is desired.
  • AAAS 63*, 65, 69, 71 (Protasan CL 210, batch 6061703)
  • AAAS 66 (0.5% w/v chitosan, Seacure CL 313, batch 7010301)
  • AAAS 79 contains the batches AAAS 64, 67, 70, 72, 74, 75 and 76 (prepared from Seacure CL 313); results from elemental analysis (mean of two determinations): S: 11.3, N: 3.3 (including about 20% w/w of free SOS determined as described below).
  • AAAS 82 contains the batches AAAS 65, 69, 71, 76 and 77 (prepared from Protasan CL 210); results from elemental analysis (mean of two determinations): S: 11.8, N: 3.2
  • AAAS 85-86 Provides asan CL 210, batch 7032401.
  • AAAS 102-113 (subjected to the washing procedure described below; Protasan CL 210, batch
  • Method IV 500 ml of chitosan solution (10 mg/ml) is added slowly and dropwise by a pump (Watson-Marlow), rate 10) through a tin (Finn 5-50 ⁇ l) to 2000 ml of SOS solution
  • chitosan solution (12.5 mg/ml) at room temperature.
  • the 500 ml of chitosan solution is added over a time period of about 3 hours.
  • Small transparent spheres containing the chitosan solution are formed.
  • the obtained mixture is allowed to react for 4 hours from start of the experiment.
  • the chitosan solution reacts with SOS on the surface of the drops and small spheres are formed and filtered off using a paper filter.
  • the chitosan-SOS formed is subjected to a washing procedure in order to remove any unbound or loose bound water-soluble SOS.
  • the washing procedure involves suspending of the chitosan-SOS in about 200 ml of distilled water while stirring. The liquid is filtered off and the material left on the filter is slowly poured over with approximately 200 ml of water twice. The remaining material is dried in an oven at
  • the chitosan-SOS formed is subjected to a washing procedure in order to remove any unbound or loose bound water-soluble SOS.
  • the washing procedure involves suspending of the chitosan-SOS in about 200 ml of distilled water for 15 min while stirring. The liquid is filtered off and the procedure is repeated once. After filtering, the material left on the filter is slowly poured over with approximately 100 ml of water twice. The remaining material is dried in an oven at 60°C for 2 hours. Dried flakes are obtained.
  • the products obtained are clear and very hard flakes having a pale yellowish colour. These flakes can only with difficulties be pulverized to small particles by means of a mortar.
  • a white yellowish powder is obtained which is free-flowing.
  • the mean particle size (AAAS 79 and AAAS 82) is determined to 50-60 ⁇ m by use of Coulter Multisizer II and employing 0.9% sodium chloride and 0.1% Tween 80 as an electrolyte solution.
  • Chitosan solution A 1% w/v (10 mg/ml) chitosan solution is prepared by slurring 5.0 g chitosan in 250 ml of distilled water followed by adding 250 ml of a 2% w/v solution of acetic acid. The resulting mixture, i.e. a 1% w/v solution of chitosan in 1% w/v acetic acid, is stirred vigorously for 60 min or until dissolution has taken place. Alternatively, the chitosan can be dissolved directly in 1% acetic acid solution.
  • SOS solution 1, 2.5, 5 and 25 g sucrose octasulfate sodium salt (SOS) is added to 2000 ml deionized water containing 0.5, 1.25, 2.5 and 12.5 mg/ml of SOS. After isolation and washing of the product obtained it is dried and the yield is determined. Elemental analysis is also performed and the results are the following:
  • the products obtained were subjected to testing for free, unbound or loose bound SOS.
  • 20 ml of distilled water is added to 100 mg of the product obtained (concentration 5 mg/ml).
  • the mixture is magnetically stirred at room temperature.
  • time t 1 min, 30 min, 2 hours, 24 hours a 2 ml sample is withdrawn and filtered.
  • the filtrate is analyzed for SOS. None of the samples analyzed from batches BHQ01-03 contained any free SOS.
  • the stoechiometric composition of the product (the combination) is independent of the batch size the product obtained can be produced in a reproducible manner
  • a product made according to the general method described above contains about 12% sulfur and 3% nitrogen corresponding to about 45% SOS and about 55% chitosan (including loose bound and unbound SOS) the use of different qualities of chitosan with the same degree of deacetylation corresponding to about 84% (Protasan CL 210 and Seacure CL 313) as starting materials does not result in a significant difference in the products obtained with respect to the content of S and N * employment of the washing procedure described above is an advantage in order to remove any loose bound or unbound SOS from the products (compare the results from the elemental analysis of AAAS 114 (washed) and AAAS 82 (unwashed)).
  • compositions and the yield of the combination obtained such as, e.g., the method of preparation (e.g. the mixing process, the mixing process and the order in which the components is added, the ionic strength prevailing in the reaction mixture, the viscosity, the kind of solvent chosen for the reactants, any addition of accelerators, the ratio of SOS and chitosan, the pH prevailing in the reaction mixture, reaction time, rotation speed, e.g. from 0-1000 rpm, homogenisation of the reaction mixture, heating/cooling of the reaction mixture, employment of ultrasonic treatment (cf.
  • the method of preparation e.g. the mixing process, the mixing process and the order in which the components is added, the ionic strength prevailing in the reaction mixture, the viscosity, the kind of solvent chosen for the reactants, any addition of accelerators, the ratio of SOS and chitosan, the pH prevailing in the reaction mixture, reaction time, rotation speed, e.g. from 0-1000 rpm, homogenisation of the reaction mixture
  • Example 5 the starting materials (chitosan, chitosan glutamate, chitosan chloride, chitosan lactate, sucroseoctasulfate as hydrate or other solvates or as a sodium, potassium or another salt) and the concentration thereof (the concentration of chitosan may be varied from about 0.1% to about 20% w/v and the concentration of SOS may be varied from about 0.1% to about 50% w/v), the precipitation method, any purification step etc.). Investigations are still ongoing with regard to testing relevant process parameters.
  • a chitosan-sodium sucroseoctasufate film can also be obtained starting from pouring a sodium sucroseoctasulfate solution into a petri dish and then adding a chitosan solution thereto.
  • thermograms obtained are enclosed (see Fig. 1-4) and they show that i) chitosan decomposes at about 230°C; ii) sodium sucroseoctasulfate has a sharp endotherm at about 100°C probably indicating a melting of sodium sucroseoctasulfate hydrate.
  • the starting material for all experiments given herein has been sodium sucroseoctasulfate hydrate, iii) sodium sucrose octasulfate decomposes at about 150°C; iv) the physical blend of chitosan and sodium sucroseoctasulfate shows a sharp endotherm at about 100°C, a decomposition of sucrose octasulfate at about 150°C and a decomposition of chitosan of about 230°C; v) the .AAAS-5-2, i.e.
  • thermogram for the combination does not have any endotherm at 100°C nor shows it any decomposition of sodium sucrose octasulfate and chitosan at about 150°C and about 230°C, respectively.
  • the AAAS-5-2 i.e. the chitosan-sodium sucrose octasulfate combination
  • the solubility was determined visually by adding 5 mg of batch AAAS-5-2 (pulverized in a mortar) to 20 ml of the medium under investigation. The mixture was then heated to about 50°C on a ultrasonic bath for 30 min.
  • Solubility of sodium sucrose octasulfate in the above-mentioned media soluble in media 1, 2, 3, 4 and 6 and only sparingly soluble in media 5 (less than 0.025 % w/v, i.e. less than 0.25 mg/ml).
  • SOS-Na 8 is also soluble in 1% w/v acetic acid.
  • Solubility of chitosan (as base) in the above mentioned media As mentioned above, chitosan is not soluble at pH above 7.0; therefore, to avoid precipitation, the pH must be maintained at pH 6.0 or below. Generally the solubility of chitosan in inorganic acids is as follows:
  • HC1/HN0 3 Chitosan is soluble in a 0.15-1.1 % solution of the acid but insoluble in a 10 % solution of the acid.
  • H 2 S0 4 not soluble at any concentration.
  • H 3 P0 Only slightly soluble in a 0.5% solution of the acid.
  • a preferred method to make a 1 % chitosan solution is to slurry the chitosan in water and then add the solution of the desired organic acid (normally acetic acid). This mixture should be stirred vigorously for 60 minutes or until complete solubility is realized.
  • chitosan can be dissolved directly by adding chitosan to a prepared 1 % solution of the organic acid.
  • the solubility may also be determined by trituration.
  • An accurate amount of the compound/material under testing is loaded into a tube and then an accurate volume (e.g. from 10 to 500 ⁇ l) of the dissolving medium is added.
  • the mixture is stirred at a constant temperature and after a fixed period of time e.g. from 1 to 24 hours, the mixture is visually inspected and if solid particles still are present then an additional volume of the medium is added. This procedure is repeated until dissolution of the compound/material is completed.
  • the solubility is then calculated taking into account the accurate amount of the compound/material tested and the total volume of the medium used for dissolution.
  • solubility can be determined at e.g. 22°C using a series of different concentration ranges of the combination. The test is run for e.g. 24 hours or longer, if necessary.
  • aqueous solubility of chitosan-SOS was determined in deionized water and 0.9% w/v sodium chloride solution, respectively.
  • the solubilities were examined at room temperature (about 22°C) by adding excess amounts of the compound to the solvents in tightly closed containers.
  • the mixture were placed in an ultrasonic water hath for about 30 min and then rotated on a mechanical spindel for 2-3 days. It was ensured that saturation equilibrium was established.
  • a concentration range of chitosan-SOS in the solvents was examined. The solubility was assessed by visually inspection of undissolved particles in the glass containers.
  • aqueous solubility of chitosan-SOS premilled ⁇ AS 79 in both deionized water and in 0.9% w/v sodium chloride solution, respectively, is less than 0.001 mg/ml (1 ⁇ g/ml).
  • solubility of chitosan-SOS in 1 M aqueous ammoniumchloride (pK A 9.25) pH 5.0 has been found to be less than 0.01 mg/ml.
  • Lysozyme is an enzyme which is present in wounds of mammals and, furthermore, Lysozyme L 6876 from Sigma is able to hydrolyze chitosan to glucosamine residues.
  • the aim of the study was to investigate whether the influence of lysozyme on a chitosan-sodium sucrose octasulfate combination releases sodium sucrose octasulfate from the combination.
  • AAAS-5-2 i.e. a chitosan-sodium sucroseoctasulfate combination
  • AAAS-5-2 i.e. a chitosan-sodium sucroseoctasulfate combination
  • the amount of sodium sucrose octasulfate released was determined by employment of the HPLC assay described in USP 23 (1995) page 1443 for sucralfate.
  • glucosidases various glucosidases, proteases, elastase, chitonase) and iii) at conditions where the enzyme used have its optimum activity (e.g.lysozyme-activity is very dependent on the pH of the solution and has its the maximum activity at about pH 5).
  • AAAS 19 was made according to method I (see Example 4) and the concentration of sodium sucrose octasulfate was 12.5 mg/ml.
  • the content of SOS in batch AAAS5-2 and AAAS19 is calculated to be about 33 % w/w and 37 % w/w, respectively.
  • the elemental analysis can be used to support the results obtained from other experiments concerning determination of the SOS-content in the combination.
  • the difference in molecular structure between the three types of chitosan can explain the difference in pH (especially the deacetylation grade of the chitosan may be of importance here).
  • X-ray diffraction The Debye-Scherrer method is employed: A monochromatic X-ray beam is diffracted by a powder sample. The crystallities give rise to cones of intensity which are detected by a photographic film wrapped around the circumference of the camera.
  • the chitosan-SOS in the combination may be based on the following structures:
  • the aim of the study is to document a potential benefit of using a combination of chitosan-SOS in the healing of skin wounds.
  • Twenty patients with preferably two split donor sites receive randomly selected between the two donor sites, traditional wound dressing (control) or traditional wound dressing plus chitosan- SOS combination (test). Wound dressings are changed every two days and time to complete healing of the donor site is recorded.
  • leg ulcers receive traditional treatment (control) or traditional treatment plus a chitosan-SOS combination or alternatively, leg ulcers receive placebo (control) or treatment with a combination according to the invention.
  • Primary end point is "ulcer closure rate" calculated after measuring ulcer area weekly. Secondary end points are time to healing and overall number of completely healed ulcers in the two study groups.
  • a dose-finding study is normally carried out in order to establish the effective concentration of the active ingredient.
  • Such studies may be performed according to established guidelines for good clinical practice (GLP) or Investigational New Drug Exemption regulations as described in the description.
  • GLP clinical practice
  • Investigational New Drug Exemption regulations as described in the description.
  • the objective of the present study is to assess the tissue reactions in full thickness wounds in pigs after treatment with the following four test samples: TA-1: chitosan
  • TA-2 sodium sucrose octasulfate
  • TA-3 a combination of chitosan-sucrose octasulfate according to the invention
  • TA-4 a physical blend of chitosan and sodium sucrose octasulfate
  • TA-5 no treatment
  • test samples may be applied as solid material with or without any addition of excipient(s) or they may be applied in a vehicle
  • Tegaderm® ii) vehicle corresponding to vehicles employed for the test samples, iii) blank.
  • the substances may also be applied in the form of a gel preparation such as a 2.6% w/w Natrosol 250 HX pharm in distilled water containing the test substance. Gels containing 5-40% w/w chitosan-SOS was prepared by slowly adding the polymer powder to the water phase heated to about 80-90°C. The mixture was stirred until a homogeneous clear colourless solution was obtained.
  • chitosan-SOS combination (AAAS 82) was added to the gel solution.
  • the mixtures were mixed with a mortar and pestle. A homogeneous grey-beige coloured suspension was obtained.
  • the suspension gel does not sediment during storage for a week at 5°C.
  • Pigs have been chosen as experimental animals because pigs have proven to be a good model for assessment of wound healing in humans.
  • the study will take place in an animal room provided with filtered air at a temperature of 21°C ⁇ 3°C, relative humidity of 55% ⁇ 15% and an air change of 10 times/hour.
  • the room will be illuminated to give a cycle of 12 hours light and 12 hours darkness.
  • Light will be on from 6 a.m. to 6 p.m.
  • the pigs will be housed individually in metal cages.
  • a commercially available pelleted mini-pig diet, Altromin 9023 from Chr. Petersen A/S, Ringsted, Denmark will be offered (400 g per pig twice daily). Analyses for nutritional components and relevant possible contaminants are performed regularly. Certificates of analysis are retained. A supply of autoclaved hay will be given daily. Twice daily the pigs will be offered domestic quality drinking water ad libitum. Analyses for relevant possible contaminants are performed regularly. Certificates of analysis are retained.
  • each pig On the day of arrival, the animals will be assigned the numbers 1, 2, 3, 4 and 5 by a randomization procedure using a randomization scheme. Each pig will be identified by an individual number tagged to the pinna of one ear. In addition each pig will be ascribed a cage card which identifies the pig by animal number and study number.
  • the pigs On the first day (day 0) the pigs will be anaesthetized with Stressimil® (i.m.) and Hypnodil® (i.v.). Atropin (i.m.) will be given at the same time to prevent salication.
  • the hairs of the back and flank will be clipped with an electric clipper before washing with soap and water followed by disinfection with 70% ethanol. Before surgery the skin of the back will be rinsed with sterile 0.9% saline.
  • Circular full thickness wounds will be prepared surgically (diameter 20 mm). Ten wounds (5 on each site of the midline) will be prepared on each pig. A schematic illustration of number and localization of wounds are given below:
  • Wound dressings are as follows:
  • each wound will be traced i.e. a drawing of the margin of the wound on a sterile transparent film using a thin pencil.
  • the epithelial rim will also be traced.
  • each of the test dressings will be applied to the wounds.
  • the allocation of treatments is described above. This treatment scheme secures and even distribution of the treatments to the different wound positions spread over the five pigs.
  • the wound can be covered with Tagaderm and thereafter gauze (4 layers) which will be fixed with 2.5 cm Scanpor Tape. Subsequently, the test area will be covered/fixed with Fizomull. Finally the pig will be dressed in Fixonet elasticated net dressing. Every day or every second day the applied dressings will be removed and new dressings and drug composition will be applied according to the same procedure used on day 0. The pigs are placed in a hammmock in order to facilitate the procedure.
  • the animals will be weighed on arrival, on the day of wounding and on the day of termination of the study.
  • the animals will be inspected at least once daily for signs of ill health.
  • the dressings will be inspected and any detached dressing reapplied or, if necessary, redressed with the same type of dressing.
  • Each wound will be observed and evaluated daily. The grade of inflammation and exudation will be evaluated. Special attention will be paid to occurrence of non-degraded test article in the wound (occurrence of visible particles in the wound). The wound edge diameter and the epithelial edge diameter will be measured. Photographs of each wound will be taken daily. All observations will be recorded.
  • each wound will be excised with a full thickness cut and at least a 5 mm margin of normal skin around the wound.
  • Each wound will be attached to a piece of cardboard with one piece of gauze in between and fixed in neutral 4% buffered formaldehyde. Care will be taken in order to secure that animal number, wound number and position can be identified.
  • sample 1 and 2 Two tissue samples (sample 1 and 2) from each wound will be paraffin embedded and sections cut at nominal thickness of 5 ⁇ m will be stained with haematoxylin - eosin. The slides will be examined microscopically (slide 1 and 2).
  • tissue will represent two full transversal (medio-laterally) sections of each wound and surrounding skin.
  • each tracing sheet is made and the area of each wound and epithelial rim is determined by video planimetry, using a video camera (Kafpa CF8) and an Image Analysis System Cream (Kem-En-Tec software Systems) version 4.0.
  • the concentration of the combination is 10% w/w.
  • Other concentration can be obtained by adjusting the amount of the combination in the cream.
  • cream bases can also be used such as e.g. Decubal® cream (added from about 0.1% to about 40% w/w chitosan-SOS).
  • 2% w/v glycerol optionally a suitable preservative in an appropriate concentration up to 100% w/v purified water, adjusted to pH 5.6
  • chitosan 200 ml chitosan (2% w/v) was mixed with 100 ml of methyl cellulose (2% w/v) and then 8 ml of glycerol was added under stirring. 8 g chitosan-SOS combination was dispersed in the highly viscous solution and the volume adjusted to 400 ml with purified water.
  • the methylparahydroxybenzoate was dissolved in 190 ml water and then Protasan and methyl cellulose was added. The mixture was shaken for 4 hours at a shaking table at room temperature to obtain a homogeneous viscous solution. The chitosan-SOS combination (pulverized) was dispersed in the viscous phase.
  • gels have also been prepared containing chitosan-SOS combination in various concentrations (1-5% w/w).
  • the gels are based on the following polymers:
  • Chitosan-SOS suitable suspension gels are formed; up to about 40% w/w of chitosan-SOS can easily be incorporated
  • Chitosan increases the viscosity of the gel, but a suitable gel can be prepared by adjusting the concentration of chitosan and polymer
  • Chitosan + SOS (blend): Most likely chitosan-SOS is formed in an uncontrollable manner. Precipitation of chitosan-SOS has been observed. The gels obtained were not suitable for medicinal application purposes as they were inhomogeneous.
  • gels are, e.g., carboxymethylcellulose sodium, pectin, gelatin, carboxymethylcellulose and hydroxyethylcellulose (suitable if growth factors are added), and alginate pH 7 containing hyaloronate.
  • An ointment is prepared containing the following constituents:
  • An ointment is prepared containing the following constituents:
  • the ointment is made by mixing 100 ml of an aqueous dispersion of the chitosan-SOS combination (2% w/v) in a mixer with 50 ml methyl cellulose (2% w/v) 50 ml of distilled water is added into the mixer. Finally, 4 ml of glycerol is added under continued stirring. The ointment obtained has a highly viscous consistency.
  • the powder was prepared by mixing the chitosan-SOS combination with the lactose using a mortal.
  • the powder can directly be applied to the damaged tissue or it may be covered by a dressing or adhesive.
  • the powders B-D were prepared in an analogues manner to the method described under A.
  • the powders may also be covered by a dressing or an adhesive.
  • Magnesium stearate 1.0% w/w Tocopheryl acetate ( ⁇ -tocopherol acetate) 2.0% w/w

Abstract

L'invention concerne la combinaison d'un premier composé d'un oligo- ou polysaccharide, contenant des unités aminosucre, et d'un second composé d'un mono-, di- ou oligosaccharide sulfaté en tant qu'agent de promotion de la cicatrisation de blessures dans des tissus contenant du collagène, notamment la peau et les os, et dans les muqueuses. Le premier composé peut être, par exemple, du chitosane, des chitosanes obtenus par désacétylation de la chitine à divers degrés de désacétylation, des dérivés de chitosane, des glycosaminoglycanes comprenant la chondroïtine, le sulfate de chondroïtine, l'acide hyaluronique, le sulfate de dermatane et le sulfate de kératane, des dextranes aminés comprenant le dextrane-DEAE, l'amidon aminé, le glycogène aminé, la cellulose aminée, la pectine aminée, l'héparine et des sels, complexes, dérivés et mélanges de ceux-ci, et le second composé peut être un disaccharide tel que, par exemple, du sucrose, un dérivé de sucrose ou un complexe ou sel de celui-ci, le disaccharide étant au moins tétrasulfaté. On réalise une combinaison très intéressante avec du chitosane, en tant que premier composé, et de l'octasulfate de sucrose, en tant que second composé. L'invention concerne également l'utilisation de tels composés en tant que médicaments, des compositions pharmaceutiques contenant de telles combinaisons ainsi qu'un procédé de préparation de ces combinaisons.
PCT/DK1997/000525 1996-11-15 1997-11-14 Procede visant a promouvoir la reparation tissulaire WO1998022114A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU49412/97A AU4941297A (en) 1996-11-15 1997-11-14 A method for promoting tissue repair

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
DK129796 1996-11-15
DK1297/96 1996-11-15
US3544497P 1997-01-30 1997-01-30
US60/035,444 1997-01-30

Publications (1)

Publication Number Publication Date
WO1998022114A1 true WO1998022114A1 (fr) 1998-05-28

Family

ID=26065547

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK1997/000525 WO1998022114A1 (fr) 1996-11-15 1997-11-14 Procede visant a promouvoir la reparation tissulaire

Country Status (2)

Country Link
AU (1) AU4941297A (fr)
WO (1) WO1998022114A1 (fr)

Cited By (30)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0965330A1 (fr) 1998-06-13 1999-12-22 Beiersdorf Aktiengesellschaft Compositions cosmétiques et dermatologiques contenant du chitosane et des phospholipides
WO2000056275A1 (fr) * 1999-03-22 2000-09-28 Virbac Compositions a base d'un sulfate de chondroïtine et de chitosan pour prevenir ou traiter les affections rhumatologiques par voie generale
EP1083906A1 (fr) * 1998-06-04 2001-03-21 Nutramax Laboratories, Inc. Composition a base d'aminosucre, de glycosaminoglycane et de s-adenosylmethionine pour le traitement et la reparation des tissus conjonctifs
US6372257B1 (en) 1999-06-29 2002-04-16 J. Alexander Marchosky Compositions and methods for forming and strengthening bone
WO2003035656A1 (fr) * 2001-10-19 2003-05-01 Interalia S.R.L. Sels d'argent de saccharose-octasulfate
WO2003055536A1 (fr) * 2001-12-21 2003-07-10 Coloplast A/S Dispositif de traitement de plaie
WO2004110511A1 (fr) * 2003-06-19 2004-12-23 Coloplast A/S Dispositif de soin des plaies
EP1539132A1 (fr) * 2002-07-25 2005-06-15 Advanced Life Sciences, Inc. Utilisation des acides 2,3 alkylcarbonyloxybenzoiques, de leurs derives et analogues pour le traitement des tissus et de dysfonctionnements, alterations et lesions cellulaires chez les mammiferes
EP1609462A1 (fr) * 2004-04-22 2005-12-28 JUVENA (International) AG Préparation cosmétique ou dermatologique comprenant un milieu nutritif
US7148209B2 (en) 2000-06-29 2006-12-12 Ecole Polytechnique Composition and method for the repair and regeneration of cartilage and other tissues
WO2007036044A1 (fr) * 2005-09-28 2007-04-05 Dnp Canada Inc. Combinaison de polychitosamine et de fibrate pour prevenir et traiter l'hyperlipidemie
WO2009028965A1 (fr) 2007-08-28 2009-03-05 Theodore Athanasiadis Hydrogel chirurgical
ES2327480A1 (es) * 2007-06-15 2009-10-29 Bioiberica, S.A. "disacaridos para el tratamiento de tendones, ligamentos y huesos".
US7928069B2 (en) 2002-06-21 2011-04-19 University Of Utah Research Foundation Crosslinked compounds and methods of making and using thereof
US20120108509A1 (en) * 2010-10-27 2012-05-03 Medtronic, Inc. Artificial scab for use in an airway
CN102440959A (zh) * 2011-11-29 2012-05-09 海南灵康制药有限公司 一种匹多莫德脂质体固体制剂
US8198268B2 (en) 2008-10-31 2012-06-12 Janssen Biotech, Inc. Tianeptine sulfate salt forms and methods of making and using the same
US8518422B2 (en) 2003-05-24 2013-08-27 La Prairie Group Ag Cosmetic or dermatological preparation comprising a nutrient medium phase
US8563040B2 (en) 2002-02-07 2013-10-22 Marfly 2, Lp Compositions and methods for forming and strengthening bone
WO2014009488A2 (fr) 2012-07-13 2014-01-16 Laboratoires Urgo Pansement a liberation prolongee d'actifs
US8920842B2 (en) 1999-11-15 2014-12-30 Piramal Healthcare (Canada) Ltd. Temperature controlled and pH dependent self gelling biopolymeric aqueous solution
US8927491B2 (en) 2010-12-28 2015-01-06 Depuy Mitek, Llc Methods for forming compositions for treating joints comprising bone morphogenetic protein and hyaluronic acid
US9314530B2 (en) 2012-06-13 2016-04-19 Laboratoires Vivacy Composition, in aqueous medium, that comprises at least a hyaluronic acid and at least an hydrosoluble salt of sucrose octasulfate
FR3044555A1 (fr) * 2015-12-07 2017-06-09 Urgo Lab Film comprenant un oligosaccharide polysulfate et un polycation et son procede de fabrication
US9682099B2 (en) 2015-01-20 2017-06-20 DePuy Synthes Products, Inc. Compositions and methods for treating joints
IT201700047632A1 (it) * 2017-05-03 2018-11-03 Ricerfarma Srl Composizioni topiche per mantenere e ripristinare l'omeostasi idrica della pelle
US10865383B2 (en) 2011-07-12 2020-12-15 Lineage Cell Therapeutics, Inc. Methods and formulations for orthopedic cell therapy
US11090328B2 (en) 2010-12-28 2021-08-17 Medos International Sarl Compositions and methods for treating joints
CN114028345A (zh) * 2021-11-18 2022-02-11 海南海灵化学制药有限公司 一种注射用阿扑西林冻干剂及其制备工艺
CN114887121A (zh) * 2022-04-14 2022-08-12 广东粤港澳大湾区国家纳米科技创新研究院 青蒿琥酯-纳米金复合物的胶体液及其制备方法和抗菌产品

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0427190A2 (fr) * 1989-11-09 1991-05-15 Hoechst Aktiengesellschaft Complexes de polyélectrolytes pour le traitement et la prophylaxie de maladies virales et leur procédé de préparation
EP0454044A2 (fr) * 1990-04-25 1991-10-30 Hoechst Aktiengesellschaft Produit pharmacologique contenant des composés de polyélectrolytes sous forme de microparticules et au moins un agent actif
EP0486998A1 (fr) * 1990-11-20 1992-05-27 Unitika Ltd. Composition à libération retardée à base d'agents anticancéreux contenant du platine
WO1995030403A1 (fr) * 1994-05-04 1995-11-16 Medicarb Ab Composition d'hygiene buccale

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0427190A2 (fr) * 1989-11-09 1991-05-15 Hoechst Aktiengesellschaft Complexes de polyélectrolytes pour le traitement et la prophylaxie de maladies virales et leur procédé de préparation
EP0454044A2 (fr) * 1990-04-25 1991-10-30 Hoechst Aktiengesellschaft Produit pharmacologique contenant des composés de polyélectrolytes sous forme de microparticules et au moins un agent actif
EP0486998A1 (fr) * 1990-11-20 1992-05-27 Unitika Ltd. Composition à libération retardée à base d'agents anticancéreux contenant du platine
WO1995030403A1 (fr) * 1994-05-04 1995-11-16 Medicarb Ab Composition d'hygiene buccale

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
S. SHIRAISHI ET AL.: "controlled release of indomethacin by chitosan-polyelectrolyte complex: optimization and in vivo/in vitro evaluation", JOURNAL OF CONTROLLED RELEASE, vol. 25, no. 3, June 1993 (1993-06-01), AMSTERDAM (NL), pages 217 - 225, XP000369908 *
THO NGUYEN-XUAN ET AL.: "mucoadhesive semi-solid formulations for intraoral use containing sucralfate", EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS, vol. 42, no. 6, April 1996 (1996-04-01), STUTTGART (DE), pages 133 - 137, XP000582519 *

Cited By (54)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9254301B2 (en) 1998-03-30 2016-02-09 Marfly2, LP Compositions and methods for forming and strengthening bone
EP1083906A1 (fr) * 1998-06-04 2001-03-21 Nutramax Laboratories, Inc. Composition a base d'aminosucre, de glycosaminoglycane et de s-adenosylmethionine pour le traitement et la reparation des tissus conjonctifs
EP1083906A4 (fr) * 1998-06-04 2003-03-26 Nutramax Lab Inc Composition a base d'aminosucre, de glycosaminoglycane et de s-adenosylmethionine pour le traitement et la reparation des tissus conjonctifs
EP0965330A1 (fr) 1998-06-13 1999-12-22 Beiersdorf Aktiengesellschaft Compositions cosmétiques et dermatologiques contenant du chitosane et des phospholipides
WO2000056275A1 (fr) * 1999-03-22 2000-09-28 Virbac Compositions a base d'un sulfate de chondroïtine et de chitosan pour prevenir ou traiter les affections rhumatologiques par voie generale
US6599888B1 (en) 1999-03-22 2003-07-29 Virbac Chondroitin sulphate and chitosan compositions for treating rheumatic disorders
US6372257B1 (en) 1999-06-29 2002-04-16 J. Alexander Marchosky Compositions and methods for forming and strengthening bone
US8920842B2 (en) 1999-11-15 2014-12-30 Piramal Healthcare (Canada) Ltd. Temperature controlled and pH dependent self gelling biopolymeric aqueous solution
US7148209B2 (en) 2000-06-29 2006-12-12 Ecole Polytechnique Composition and method for the repair and regeneration of cartilage and other tissues
WO2003035656A1 (fr) * 2001-10-19 2003-05-01 Interalia S.R.L. Sels d'argent de saccharose-octasulfate
US7183315B2 (en) 2001-10-19 2007-02-27 Interalia S.R.L. Silver salts of sucrose-octasulphate
EP1576969A1 (fr) * 2001-12-21 2005-09-21 Coloplast A/S Dispositif de traitement de plaie
AU2002367083B2 (en) * 2001-12-21 2008-02-28 Coloplast A/S A wound care device
US8790688B2 (en) 2001-12-21 2014-07-29 Coloplast A/S Wound care device for local treatment of pain in a wound
WO2003055536A1 (fr) * 2001-12-21 2003-07-10 Coloplast A/S Dispositif de traitement de plaie
US8563040B2 (en) 2002-02-07 2013-10-22 Marfly 2, Lp Compositions and methods for forming and strengthening bone
US7928069B2 (en) 2002-06-21 2011-04-19 University Of Utah Research Foundation Crosslinked compounds and methods of making and using thereof
US8859523B2 (en) 2002-06-21 2014-10-14 University Of Utah Research Foundation Crosslinked compounds and methods of making and using thereof
EP1539132A1 (fr) * 2002-07-25 2005-06-15 Advanced Life Sciences, Inc. Utilisation des acides 2,3 alkylcarbonyloxybenzoiques, de leurs derives et analogues pour le traitement des tissus et de dysfonctionnements, alterations et lesions cellulaires chez les mammiferes
EP1539132A4 (fr) * 2002-07-25 2007-10-31 Advanced Life Sciences Inc Utilisation des acides 2,3 alkylcarbonyloxybenzoiques, de leurs derives et analogues pour le traitement des tissus et de dysfonctionnements, alterations et lesions cellulaires chez les mammiferes
US8518422B2 (en) 2003-05-24 2013-08-27 La Prairie Group Ag Cosmetic or dermatological preparation comprising a nutrient medium phase
US8039017B2 (en) 2003-06-19 2011-10-18 Coloplast A/S Wound care device
WO2004110511A1 (fr) * 2003-06-19 2004-12-23 Coloplast A/S Dispositif de soin des plaies
US8409610B2 (en) 2003-06-19 2013-04-02 Coloplast A/S Wound care device
EP2279763A1 (fr) * 2003-06-19 2011-02-02 Coloplast A/S Dispositif de traitement des plaies
EP1609462A1 (fr) * 2004-04-22 2005-12-28 JUVENA (International) AG Préparation cosmétique ou dermatologique comprenant un milieu nutritif
WO2007036044A1 (fr) * 2005-09-28 2007-04-05 Dnp Canada Inc. Combinaison de polychitosamine et de fibrate pour prevenir et traiter l'hyperlipidemie
ES2327480A1 (es) * 2007-06-15 2009-10-29 Bioiberica, S.A. "disacaridos para el tratamiento de tendones, ligamentos y huesos".
WO2009028965A1 (fr) 2007-08-28 2009-03-05 Theodore Athanasiadis Hydrogel chirurgical
US8809301B2 (en) 2007-08-28 2014-08-19 Adelaide Research & Innovation Pty Ltd Surgical hydrogel
US8198268B2 (en) 2008-10-31 2012-06-12 Janssen Biotech, Inc. Tianeptine sulfate salt forms and methods of making and using the same
US9700648B2 (en) * 2010-10-27 2017-07-11 Medtronic, Inc. Artificial scab for use in an airway
US10279072B2 (en) 2010-10-27 2019-05-07 Medtronic, Inc. Artificial scab for use in an airway
US20120108509A1 (en) * 2010-10-27 2012-05-03 Medtronic, Inc. Artificial scab for use in an airway
US11090328B2 (en) 2010-12-28 2021-08-17 Medos International Sarl Compositions and methods for treating joints
US8927491B2 (en) 2010-12-28 2015-01-06 Depuy Mitek, Llc Methods for forming compositions for treating joints comprising bone morphogenetic protein and hyaluronic acid
US9561260B2 (en) 2010-12-28 2017-02-07 Depuy Mitek, Llc Compositions for treating joints comprising bone morphogenetic protein and hyaluronic acid
US10865383B2 (en) 2011-07-12 2020-12-15 Lineage Cell Therapeutics, Inc. Methods and formulations for orthopedic cell therapy
CN102440959A (zh) * 2011-11-29 2012-05-09 海南灵康制药有限公司 一种匹多莫德脂质体固体制剂
US9314530B2 (en) 2012-06-13 2016-04-19 Laboratoires Vivacy Composition, in aqueous medium, that comprises at least a hyaluronic acid and at least an hydrosoluble salt of sucrose octasulfate
US10172972B2 (en) 2012-07-13 2019-01-08 Laboratoires Urgo Dressing having sustained release of active agents
WO2014009488A2 (fr) 2012-07-13 2014-01-16 Laboratoires Urgo Pansement a liberation prolongee d'actifs
US10532069B2 (en) 2015-01-20 2020-01-14 DePuy Synthes Products, Inc. Compositions and methods for treating joints
US9682099B2 (en) 2015-01-20 2017-06-20 DePuy Synthes Products, Inc. Compositions and methods for treating joints
US20180353333A1 (en) * 2015-12-07 2018-12-13 Urgo Recherche Innovation Et Developpement Film Including a Polysulfated Oligosaccharide and a Polycation and Method for Manufacturing Same
WO2017098118A1 (fr) 2015-12-07 2017-06-15 Urgo Recherche Innovation Et Developpement Film comprenant un oligosaccharide polysulfaté et un polycation et son procédé de fabrication
CN108366972A (zh) * 2015-12-07 2018-08-03 国家科学研究中心 包括多硫酸化寡糖和聚阳离子的膜及其制造方法
FR3044555A1 (fr) * 2015-12-07 2017-06-09 Urgo Lab Film comprenant un oligosaccharide polysulfate et un polycation et son procede de fabrication
CN108366972B (zh) * 2015-12-07 2021-03-30 国家科学研究中心 包括多硫酸化寡糖和聚阳离子的膜及其制造方法
WO2018202456A1 (fr) * 2017-05-03 2018-11-08 Ricerfarma S.R.L. Compositions topiques conçues pour maintenir et restaurer l'homéostasie de l'eau de la peau
IT201700047632A1 (it) * 2017-05-03 2018-11-03 Ricerfarma Srl Composizioni topiche per mantenere e ripristinare l'omeostasi idrica della pelle
CN114028345A (zh) * 2021-11-18 2022-02-11 海南海灵化学制药有限公司 一种注射用阿扑西林冻干剂及其制备工艺
CN114028345B (zh) * 2021-11-18 2023-02-28 海南海灵化学制药有限公司 一种注射用阿扑西林冻干剂及其制备工艺
CN114887121A (zh) * 2022-04-14 2022-08-12 广东粤港澳大湾区国家纳米科技创新研究院 青蒿琥酯-纳米金复合物的胶体液及其制备方法和抗菌产品

Also Published As

Publication number Publication date
AU4941297A (en) 1998-06-10

Similar Documents

Publication Publication Date Title
WO1998022114A1 (fr) Procede visant a promouvoir la reparation tissulaire
KR930003117B1 (ko) 황산화 이당류를 활성성분으로 함유하는 염증예방 또는 치료용 제약조성물
RU2241489C2 (ru) Композиции матриксных протеинов для залечивания ран
EP2076246B1 (fr) Formation de gels médicalement utiles comprenant des particules microporeuses et leurs procédés d'utilisation
JP2002521338A (ja) 抗脂血症製剤
EP1059934B1 (fr) Compositions proteiniques matricielles de cicatrisation
JP2007526273A (ja) キセロゲルまたはフィルムの形態での局所使用のための医薬組成物および製造方法
EP2139451B1 (fr) Compositions topiques contenant du magaldrate
ZA200104742B (en) Bioadhesive antibacterial wound healing composition.
RU2701513C2 (ru) Фармацевтическая композиция, содержащая электрогидродинамически полученные волокна, композиция, имеющая улучшенное время удержания в месте применения
WO2008037262A1 (fr) Composition pharmaceutique et SON utilisations
WO2003009829A2 (fr) Systeme antiemetique, et anti mal des transports a liberation controlee de medicament
WO2002094256A1 (fr) Lysine et/ou analogues et/ou polymeres de ces derniers destines a promouvoir la cicatrisation et l'angiogenese
US20230330190A1 (en) Topical composition for improved healing of open wounds
CN114364379A (zh) 含有金属有机框架材料的用于伤口愈合的组合物
US20040087544A1 (en) Pharmaceutical compositions of topic use, applied in treatment of skin and/or mucous injuries use of compositions in treatment of skin and/or mucous injuries and use of compounds in treatment of skin and/or mucous injuries
US20090317474A1 (en) Use of polyethylene glycol in inflammation related topical disorders or diseases and wound healing
CN107446018A (zh) 促进伤口愈合的肽及其应用
JP2983335B2 (ja) ホスホマイシン及びその医薬的に許容可能な塩の局所瘢痕形成剤としての使用
EP2255819A1 (fr) Extrait d'annélide et son utilisation pour la régénération de la peau
EP2470197A1 (fr) Composition pour l absorption et l action prolongée de peptides liés à la leptine
DE60008077T2 (de) Matrixproteinzusammensetzungen um Apoptose zu induzieren
US20210379161A1 (en) Topical composition for improved healing of ocular ailments
JP2004523521A (ja) 非代謝性の糖質と高分子吸収剤とを用いた細胞再生及び／又は細胞分化の促進
Saliy et al. Development strategy of novel drug formulations for the delivery of doxycycline in the treatment of wounds of various etiologies

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AT AU AZ BA BB BG BR BY CA CH CN CU CZ CZ DE DE DK DK EE ES FI FI GB GE GH HU ID IL IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SK SL TJ TM TR TT UA UG US UZ VN YU ZW AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH KE LS MW SD SZ UG ZW AT BE CH DE DK ES FI FR

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA