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
  • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
  • A61L31/08—Materials for coatings
  • A61L31/10—Macromolecular materials
• • 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
  • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
  • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
  • A61L31/16—Biologically active materials, e.g. therapeutic substances
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P19/00—Drugs for skeletal disorders
  • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/10—Antimycotics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• • 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
  • A61L2300/416—Anti-neoplastic or anti-proliferative or anti-restenosis or anti-angiogenic agents, e.g. paclitaxel, sirolimus

Definitions

• This invention relates generally to polymer compositions that include a therapeutic agent (e.g., a fibrosis-inhibiting agent or an anti-infective agent), and to methods of making and using such compositions.
• a therapeutic agent e.g., a fibrosis-inhibiting agent or an anti-infective agent
• EP 0 732 109 A1 discloses a crosslinked biomaterial composition that is prepared using a hydrophobic crosslinking agent, or a mixture of hydrophilic and hydrophobic crosslinking agents.
• U.S. Patent No. 5,614,587 describes bioadhesives that comprise collagen that is crosslinked using a multifunctionally activated synthetic hydrophilic polymer.
• composition useful in the prevention of surgical adhesions comprising a substrate material and an anti-adhesion binding agent, where the substrate material may comprise collagen and the binding agent may comprise at least one tissue-reactive functional group and at least one substrate-reactive functional group.
• U.S. application Ser. No. 08/476,825, filed Jun. 7, 1995 discloses bioadhesive compositions comprising collagen crosslinked using a multifunctionally activated synthetic hydrophilic polymer, as well as methods of using such compositions to effect adhesion between a first surface and a second surface, wherein at least one of the first and second surfaces may be a native tissue surface.
• 5,874,500 describes a crosslinked polymer composition that comprises one component having multiple nucleophilic groups and another component having multiple electrophilic groups. Covalently bonding of the nucleophilic and electrophilic groups forms a three dimensional matrix that has a variety of medical uses including tissue adhesion, surface coatings for synthetic implants, and drug delivery. More recent developments include the addition of a third component having either nucleophilic or electrophilic groups, as is described in U.S. Patent No.
• compositions that contain both an anti-fibrotic agent and either a polymer or a pre-polymer, i.e., a compound that forms a polymer.
• these compositions are formed in-situ when precursors thereof are delivered to a site in the body, or a site on an implant.
• the compositions of the invention include the crosslinked reaction product that forms when two compounds (a multifunctional polynucleophilic compound and a multi-functional polyelectrophilic compound) are delivered to a site in a host (in other words, a patient) in the presence of an anti-fibrotic agent.
• the compositions of the invention also include a mixture of anti-fibrotic agent and a polymer, where the composition can be delivered to a site in a patient's body to achieve beneficial affects, e.g., the beneficial affects described herein.
• the polymers themselves are useful in various methods, including the prevention of surgical adhesions.
• the present invention provides methods for treating and/or preventing surgical adhesions.
• the surgical adhesions can be the result of, for example, spinal or neurosurgical procedures, of gynecological procedures, of abdominal procedures, of cardiac procedures, of orthopedic procedures, of reconstructive procedures, and cosmetic procedures.
• the present invention provides methods for treating or preventing inflammatory arthritis, such as osteoarthritis and rheumatoid arthritis.
• the method includes delivering to patient in need thereof an anti-fibrotic agent, optionally with a polymer.
• the present invention provides for the prevention of cartilage loss as can occur, for example after a joint injury.
• the method includes delivering to the joint of the patient in need therof an anti- fibrotic agent, optionally with a polymer.
• the present invention provides for treating hypertrophic scars and keloids.
• the method includes delivering to the scar or keloid of the patient in need thereof an anti-fibrotic agent, optionally with a polymer.
• the present invention provides a method for the treatment of vascular disease, e.g., stenosis, restenosis or atherosclerosis.
• the method includes the perivascular delivery of an anti-fibrotic agent.
• the present invention provides a method for implanting a medical device comprising: (a) infiltrating a tissue of a host where the medical device is to be, or has been, implanted with i) an anti-fibrotic agent, ii) an anti-infective agent, iii) a polymer; iv) a composition comprising an anti- fibrotic agent and a polymer, v) a composition comprising an anti-infective agent and a polymer, or vi) a composition comprising an anti-fibrotic agent, an anti-infective agent and a polymer, and (b) implanting the medical device into the host.
• the invention provides: a method for implanting a medical device comprising: (a) infiltrating a tissue of a host where the medical device is to be, or has been, implanted with an anti-fibrotic agent, and (b) implanting the medical device into the host; a method for implanting a medical device comprising: (a) infiltrating a tissue of a host where the medical device is to be, or has been, implanted with an anti-infective agent, and (b) implanting the medical device into the host; a method for implanting a medical device comprising: (a) infiltrating a tissue of a host where the medical device is to be, or has been, implanted with a polymer; and (b) implanting the medical device into the host; a method for implanting a medical device comprising: (a) infiltrating a tissue of a host where the medical device is to be, or has been, implanted with a composition comprising an anti-fibrotic agent and
• Figure 1 is a diagram showing how a cell cycle inhibitor acts at one or more of the steps in the biological pathway.
• Figure 2 is a graph showing the results for the screening assay for assessing the effect of mitoxantrone on nitric oxide production by THP-1 macrophages.
• Figure 3 is a graph showing the results for the screening assay for assessing the effect of Bay 11-7082 on TNF-alpha production by THP-1 macrophages.
• Figure 4 is a graph showing the results for the screening assay for assessing the effect of rapamycin concentration for TNF ⁇ production by THP-1 macrophages.
• Figure 5 is graph showing the results of a screening assay for assessing the effect of mitoxantrone on proliferation of human fibroblasts.
• Figure 6 is graph showing the results of a screening assay for assessing the effect of rapamycin on proliferation of human fibroblasts.
• Figure 7 is graph showing the results of a screening assay for assessing the effect of paclitaxel on proliferation of human fibroblasts.
• Figure 8 is a picture that shows an uninjured carotid artery from a rat balloon injury model.
• Figure 9 is a picture that shows an injured carotid artery from a rat balloon injury model.
• Figure 10 is a picture that shows a paclitaxel/mesh treated carotid artery in a rat balloon injury model.
• Figure 11 A schematically depicts the transcriptional regulation of matrix metalloproteinases.
• Figure 11 B is a blot which demonstrates that IL-1 stimulates AP-1 transcriptional activity.
• Figure 11C is a graph which shows that IL-1 induced binding activity decreased in lysates from chondrocytes which were pretreated with paclitaxel.
• Figure 11 D is a blot which shows that IL-1 induction increases collagenase and stromelysin in RNA levels in chondrocytes, and that this induction can be inhibited by pretreatment with paclitaxel.
• Figures 12A-H are blots that show the effect of various anti- microtubule agents in inhibiting collagenase expression.
• Figure 13 is a graph showing the results of a screening assay for assessing the effect of paclitaxel on smooth muscle cell migration.
• Figure 14 is a graph showing the results of a screening assay for assessing the effect of geldanamycin on IL-1 ⁇ production by THP-1 macrophages.
• Figure 15 is a graph showing the results of a screening assay for assessing the effect of geldanamycin on IL-8 production by THP-1 macrophages.
• Figure 16 is a graph showing the results of a screening assay for assessing the effect of geldanamycin on MCP-1 production by THP-1 macrophages.
• Figure 17 is graph showing the results of a screening assay for assessing the effect of paclitaxel on proliferation of smooth muscle cells.
• Figure 18 is graph showing the results of a screening assay for assessing the effect of paclitaxel for proliferation of the murine RAW 264.7 macrophage cell line.
• Figure 19 is a graph showing the average rank of joint scores of Hartley guinea pig knees with ACL damage treated with paclitaxel. A reduction in score indicates an improvement in cartilage score. The dose response trend is statistically significant (p ⁇ 0.02).
• Figures 20A-C are examples of cross sections of Hartley guinea pig knees of control and paclitaxel treated animals.
• Figure 20A Control speciment showing erosion of cartilage to the bone.
• Figure 20B Paclitaxel dose 1 (low dose) showing fraying of cartilage.
• Figure 20C The dose response trend is statistically significant
• FIGS 21A-F are Safranin-O stained histological slides of representative synovial tissues from na ⁇ ve (healthy) knees ( Figures 21 A and 21 D) and knees with arthritis induced by administration of albumin in Freund's complete adjuvant ( Figures 21 B and 21 C) or carrageenan ( Figures 21 E and 21 F). Arthritic knees received either control ( Figures 21 B and 21 E) or 20% paclitaxel-loaded microspheres ( Figures 21 C and 21 F). The data illustrate decreased proteoglycan red staining in arthritic knees treated with control microspheres and the proteoglycan protection properties of the paclitaxel- loaded formulation.
• Fibrosis or “scarring,” or “fibrotic response” refers to the formation of fibrous (scar) tissue in response to injury or medical intervention.
• fibrosis-inhibiting agents which inhibit fibrosis or scarring are referred to herein as "fibrosis-inhibiting agents", “fibrosis-inhibitors”, “anti-scarring agents”, and the like, where these agents inhibit fibrosis through one or more mechanisms including: inhibiting inflammation or the acute inflammatory response, inhibiting migration or proliferation of connective tissue cells (such as fibroblasts, smooth muscle cells, vascular smooth muscle cells), inhibiting angiogenesis, reducing extracellular matrix (ECM) production or promoting ECM breakdown, and/or inhibiting tissue remodeling.
• connective tissue cells such as fibroblasts, smooth muscle cells, vascular smooth muscle cells
• ECM extracellular matrix
• a body passageway e.g., a blood vessel, the gastrointestinal tract, the respiratory tract, the urinary tract, the female or male reproductive tract, the eustacian tube etc.
• stenosis narrowing
• scarring When scarring subsequently occurs to re-occlude a body passageway after it was initially successfully opened by a surgical intervention (such as placement of a medical device or implant), this is referred to as "restenosis.”
• restenosis When scarring subsequently occurs to re-occlude a body passageway after it was initially successfully opened by a surgical intervention (such as placement of a medical device or implant), this is referred to as “restenosis.”
• “Host”, “person”, “subject”, “patient” and the like are used synonymously to refer to the living being into which a device or implant of the present invention is implanted.
• Implantted refers to having completely or partially placed a device or implant within a host. A device is partially implanted when some of the device reaches, or extends to the outside of, a host.
• Inhibit fibrosis refers to the action of agents or compositions which result in a statistically significant decrease in the formation of fibrous tissue that can be expected to occur in the absence of the agent or composition.
• Anti-infective agent refers to an agent or composition which prevents microrganisms from growing and/or slows the growth rate of microorganisms and/or is directly toxic to microorganisms at or near the site of the agent. These processes would be expected to occur at a statistically significant level at or near the site of the agent or composition relative to the effect in the absence of the agent or composition.
• Inhibit infection refers to the ability of an agent or composition to prevent microorganisms from accumulating and/or proliferating near or at the site of the agent. These processes would be expected to occur at a statistically significant level at or near the site of the agent or composition relative to the effect in the absence of the agent or composition.
• “Inhibitor” refers to an agent which prevents a biological process from occurring or slows the rate or degree of occurrence of a biological process. The process may be a general one such as scarring or refer to a specific biological action such as, for example, a molecular process resulting in release of a cytokine.
• Antagonist refers to an agent which prevents a biological process from occurring or slows the rate or degree of occurrence of a biological process. While the process may be a general one, typically this refers to a drug mechanism where the drug competes with a molecule for an active molecular site or prevents a molecule from interacting with the molecular site. In these situations, the effect is that the molecular process is inhibited.
• Ant refers to an agent which stimulates a biological process or rate or degree of occurrence of a biological process. The process may be a general one such as scarring or refer to a specific biological action such as, for example, a molecular process resulting in release of a cytokine.
• Anti-microtubule agents should be understood to include any protein, peptide, chemical, or other molecule which impairs the function of microtubules, for example, through the prevention or stabilization of polymerization.
• Compounds that stabilize polymerization of microtubules are referred to herein as "microtubule stabilizing agents.”
• a wide variety of methods may be utilized to determine the anti-microtubule activity of a particular compound, including for example, assays described by Smith et al. (Cancer Lett 7*9(2):213-219, 1994) and Mooberry et al., (Cancer Lett. 96(2):261- 266, 1995).
• Medical device “implant”, “”device”, medical device”, “medical implant”, “implant/device” and the like are used synonymously to refer to any object that is designed to be placed partially or wholly within a patient's body for one or more therapeutic or prophylactic purposes such as for restoring physiological function, alleviating symptoms associated with disease, delivering therapeutic agents, and/or repairing, replacing, or augmenting etc. damaged or diseased organs and tissues.
• some medical devices and implants include materials derived from animals (e.g., "xenografts” such as whole animal organs; animal tissues such as heart valves; naturally occurring or chemically-modified molecules such as collagen, hyaluronic acid, proteins, carbohydrates and others), human donors (e.g., "allografts” such as whole organs; tissues such as bone grafts, skin grafts and others), or from the patients themselves (e.g., "autografts” such as saphenous vein grafts, skin grafts, tendon/ligament/muscle transplants).
• animals e.g., "xenografts” such as whole animal organs; animal tissues such as heart valves; naturally occurring or chemically-modified molecules such as collagen, hyaluronic acid, proteins, carbohydrates and others
• human donors e.g., "allografts” such as whole organs; tissues such as bone grafts, skin grafts and others
• autografts such as sap
• Chondroprotection refers to the prevention of cartilage loss. Cartilage is formed from chondrocytes, and chondroprotection is the protection of the chrondrocytes so that they do not die.
• Disease of an agent refers to a statistically significant presence of the agent, or a subcomponent thereof, which has disassociated from the implant/device and/or remains active on the surface of (or within) the device/implant.
• Biodegradable refers to materials for which the degradation process is at least partially mediated by, and/or performed in, a biological system.
• Degradation refers to a chain scission process by which a polymer chain is cleaved into oligomers and monomers.
• Chain scission may occur through various mechanisms, including, for example, by chemical reaction (e.g., hydrolysis) or by a thermal or photolytic process.
• Polymer degradation may be characterized, for example, using gel permeation chromatography (GPC), which monitors the polymer molecular mass changes during erosion and drug release.
• GPC gel permeation chromatography
• Biodegradable also refers to materials may be degraded by an erosion process mediated by, and/or performed in, a biological system.
• Erosion refers to a process in which material is lost from the bulk. In the case of a polymeric system, the material may be a monomer, an oligomer, a part of a polymer backbone, or a part of the polymer bulk.
• Erosion includes (i) surface erosion, in which erosion affects only the surface and not the inner parts of a matrix; and (ii) bulk erosion, in which the entire system is rapidly hydrated and polymer chains are cleaved throughout the matrix.
• erosion generally occurs by one of three basic mechanisms (see, e.g., Heller, J., CRC Critical Review in Therapeutic Drug Carrier Systems (1984), 1(1 ), 39- 90); Siepmann, J. et al., Adv. Drug Del. Rev.
• analogue refers to a chemical compound that is structurally similar to a parent compound, but differs slightly in composition (e.g., one atom or functional group is different, added, or removed).
• the analogue may or may not have different chemical or physical properties than the original compound and may or may not have improved biological and/or chemical activity.
• the analogue may be more hydrophilic or it may have altered reactivity as compared to the parent compound.
• the analogue may mimic the chemical and/or biologically activity of the parent compound (i.e., it may have similar or identical activity), or, in some cases, may have increased or decreased activity.
• the analogue may be a naturally or non- naturally occurring (e.g., recombinant) variant of the original compound.
• An example of an analogue is a mutein (i.e., a protein analogue in which at least one amino acid is deleted, added, or substituted with another amino acid).
• Other types of analogues include isomers (enantiomers, diasteromers, and the like) and other types of chiral variants of a compound, as well as structural isomers.
• the analogue may be a branched or cyclic variant of a linear compound.
• a linear compound may have an analogue that is branched or otherwise substituted to impart certain desirable properties (e.g., improve hydrophilicity or bioavailability).
• derivative refers to a chemically or biologically modified version of a chemical compound that is structurally similar to a parent compound and (actually or theoretically) derivable from that parent compound.
• a “derivative” differs from an "analogue” in that a parent compound may be the starting material to generate a "derivative,” whereas the parent compound may not necessarily be used as the starting material to generate an “analogue.”
• a derivative may or may not have different chemical or physical properties of the parent compound.
• the derivative may be more hydrophilic or it may have altered reactivity as compared to the parent compound.
• Derivatization i.e., modification
• a hydrogen may be substituted with a halogen, such as fluorine or chlorine, or a hydroxyl group (-OH) may be replaced with a carboxylic acid moiety (-COOH).
• derivative also includes conjugates, and prodrugs of a parent compound (i.e., chemically modified derivatives which can be converted into the original compound under physiological conditions).
• the prodrug may be an inactive form of an active agent.
• the prodrug may be converted into the active form of the compound.
• Prodrugs may be formed, for example, by replacing one or two hydrogen atoms on nitrogen atoms by an acyl group (acyl prodrugs) or a carbamate group (carbamate prodrugs). More detailed information relating to prodrugs is found, for example, in Fleisher et al., Advanced Drug Delivery Reviews 19 (1996) 115; Design of Prodrugs, H. Bundgaard (ed.), Elsevier, 1985; or H. Bundgaard, Drugs of the Future 16 (1991 ) 443.
• derivative is also used to describe all solvates, for example hydrates or adducts (e.g., adducts with alcohols), active metabolites, and salts of the parent compound.
• solvates for example hydrates or adducts (e.g., adducts with alcohols), active metabolites, and salts of the parent compound.
• the type of salt that may be prepared depends on the nature of the moieties within the compound.
• acidic groups for example carboxylic acid groups
• alkali metal salts or alkaline earth metal salts e.g., sodium salts, potassium salts, magnesium salts and calcium salts
• physiologically tolerable quaternary ammonium ions and acid addition salts with ammonia and physiologically tolerable organic amines such as, for example, triethylamine, ethanolamine or tris-(2-hydroxyethyl)amine.
• Basic groups can form acid addition salts, for example with inorganic acids such as hydrochloric acid, sulfuric acid or phosphoric acid, or with organic carboxylic acids and sulfonic acids such as acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid or p-toluenesulfonic acid.
• Compounds which simultaneously contain a basic group and an acidic group for example a carboxyl group in addition to basic nitrogen atoms, can be present as zwitterions.
• Salts can be obtained by customary methods known to those skilled in the art, for example by combining a compound with an inorganic or organic acid or base in a solvent or diluent, or from other salts by cation exchange or anion exchange.
• "Hyaluronic acid” or “HA” as used herein refers to all forms of hyaluronic acid that are described or referenced herein, including those that have been processed or chemically or physically modified, as well as hyaluronic acid that has been crosslinked (for example, covalently, ionically, thermally or physically).
• HA is a glycosaminoglycan composed of a ' linear chain of about 2500 repeating disaccharide units.
• Each disaccharide unit is composed of an N-acetylglucosamine residue linked to a glucuronic acid.
• Hyaluronic acid is a natural substance that is found in the extracellular matrix of many tissues including synovial joint fluid, the vitreous humor of the eye, cartilage, blood vessels, skin and the umbilical cord.
• Commercial forms of hyaluronic acid having a molecular weight of approximately 1.2 to 1.5 million Daltons (Da) are extracted from rooster combs and other animal sources.
• Other sources of HA include HA that is isolated from cell culture / fermentation processes. Lower molecular weight HA formulations are also available from a variety of commercial sources.
• the molecule can be of variable lengths (i.e., different numbers of repeating disaccharide units and different chain branching patterns) and can be modified at several sites (through the addition or subtraction of different functional groups) without deviating from the scope of the present invention.
• inter-react refers to the formulation of covalent bonds, noncovalent bonds, or both.
• the term thus includes crosslinking, which involves both intermolecular crosslinks and optionally intramolecular crosslinks as well, arising from the formation of covalent bonds.
• Covalent bonding between two reactive groups may be direct, in which case an atom in reactive group is directly bound to an atom in the other reactive group, or it may be indirect, through a linking group.
• Noncovalent bonds include ionic (electrostatic) bonds, hydrogen bonds, or the association of hydrophobic molecular segments, which may be the same or different.
• a crosslinked matrix may, in addition to covalent bonds, also include such intermolecular and/or intramolecular noncovalent bonds.
• hydrophilic and hydrophobic are generally defined in terms of an HLB value, i.e., a hydrophilic lipophilic balance. A high HLB value indicates a hydrophilic compound, while a low HLB value characterizes a hydrophobic compound. HLB values are well known in the art, and generally range from 1 to 18.
• Preferred multifunctional compound cores are hydrophilic, although as long as the multifunctional compound as a whole contains at least one hydrophilic component, crosslinkable hydrophobic components may also be present.
• synthetic is used to refer to polymers, compounds and other such materials that are "chemically synthesized.”
• a synthetic material in the present compositions may have a molecular structure that is identical to a naturally occurring material, but the material perse, as incorporated in the compositions of the invention, has been chemically synthesized in the laboratory or industrially.
• “Synthetic” materials also include semi-synthetic materials, i.e., naturally occurring materials, obtained from a natural source, that have been chemically modified in some way.
• the synthetic materials herein are purely synthetic, i.e., they are neither semi-synthetic nor have a structure that is identical to that of a naturally occurring material.
• the term "effective amount” refers to the amount of composition required in order to obtain the effect desired.
• a "tissue growth- promoting amount" of a composition refers to the amount needed in order to stimulate tissue growth to a detectable degree.
• Tissue in this context, includes connective tissue, bone, cartilage, epidermis and dermis, blood, and other tissues. The actual amount that is determined to be an effective amount will vary depending on factors such as the size, condition, sex and age of the patient and can be more readily determined by the caregiver.
• compositions of the invention can be injected or otherwise applied to a specific site within a patient's body, e.g., a site in need of augmentation, and allowed to crosslink at the site of injection.
• Suitable sites will generally be intradermal or subcutaneous regions for augmenting dermal support, at a bone fracture site for bone repair, within sphincter tissue for sphincter augmentation (e.g., for restoration of continence), within a wound or suture, to promote tissue regrowth; and within or adjacent to vessel anastomoses, to promote vessel regrowth.
• aqueous medium includes solutions, suspensions, dispersions, colloids, and the like containing water.
• alkyl refers to a branched or unbranched saturated hydrocarbon group typically although not necessarily containing 1 to about 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, f-butyl, octyl, decyl, and the like, as well as cycloalkyl groups such as cyclopentyl, cyclohexyl and the like.
• alkyl groups herein contain 1 to about 12 carbon atoms.
• the term “lower alkyl” intends an alkyl group of one to six carbon atoms, preferably one to four carbon atoms.
• “Substituted alkyl” refers to alkyl substituted with one or more substituent groups.
• Alkylene “lower alkylene” and “substituted alkylene” refer to divalent alkyl, lower alkyl, and substituted alkyl groups, respectively.
• aryl refers to an aromatic substituent containing a single aromatic ring (monocyclic) or multiple aromatic rings that are fused together, linked covalently, or linked to a common group such as a methylene or ethylene moiety.
• the common linking group may also be a carbonyl as in benzophenone, an oxygen atom as in diphenylether, or a nitrogen atom as in diphenylamine.
• Preferred aryl groups contain one aromatic ring or two fused or linked aromatic rings, e.g., phenyl, naphthyl, biphenyl, diphenylether, diphenylamine, benzophenone, and the like.
• Substituted aryl refers to an aryl moiety substituted with one or more substituent groups
• heteroatom-containing aryl and “heteroaryl” refer to aryl in which at least one carbon atom is replaced with a heteroatom.
• arylene and “substituted arylene” refer to divalent aryl and substituted aryl groups as just defined.
• heteroatom-containing as in a “heteroatom-containing hydrocarbyl group” refers to a molecule or molecular fragment in which one or more carbon atoms is replaced with an atom other than carbon, e.g., nitrogen, oxygen, sulfur, phosphorus or silicon.
• Hydrocarbyl refers to univalent hydrocarbyl radicals containing 1 to about 30 carbon atoms, preferably 1 to about 24 carbon atoms, most preferably 1 to about 12 carbon atoms, including branched or unbranched, saturated or unsaturated species, such as alkyl groups, alkenyl groups, aryl groups, and the like.
• lower hydrocarbyl intends a hydrocarbyl group of one to six carbon atoms, preferably one to four carbon atoms.
• hydrocarbylene intends a divalent hydrocarbyl moiety containing 1 to about 30 carbon atoms, preferably 1 to about 24 carbon atoms, most preferably 1 to about 12 carbon atoms, including branched or unbranched, saturated or unsaturated species, or the like.
• lower hydrocarbylene intends a hydrocarbylene group of one to six carbon atoms, preferably one to four carbon atoms.
• Substituted hydrocarbyl refers to hydrocarbyl substituted with one or more substituent groups
• heteroatom-containing hydrocarbyl and heterohydrocarbyl refer to hydrocarbyl in which at least one carbon atom is replaced with a heteroatom.
• substituted hydrocarbylene refers to hydrocarbylene substituted with one or more substituent groups
• heteroatom-containing hydrocarbylene and heterohydrocarbylene refer to hydrocarbylene in which at least one carbon atom is replaced with a heteroatom. If not otherwise indicated, “hydrocarbyl” indicates both unsubstituted and substituted hydrocarbyls, “heteroatom-containing hydrocarbyl” indicates both unsubstituted and substituted heteroatom- containing hydrocarbyls and so forth.
• substituted as in “substituted hydrocarbyl,” “substituted alkyl,” and the like, as alluded to in some of the aforementioned definitions, is meant that in the hydrocarbyl, alkyl, or other moiety, at least one hydrogen atom bound to a carbon atom is replaced with one or more substituents that are functional groups such as alkoxy, hydroxy, halo, nitro, and the like. Unless otherwise indicated, it is to be understood that specified molecular segments can be substituted with one or more substituents that do not compromise a compound's utility.
• sucinimidyl is intended to include unsubstituted succinimidyl as well as sulfosuccinimidyl and other succinimidyl groups substituted on a ring carbon atom, e.g., with alkoxy substituents, polyether substituents, or the like.
• concentration ranges, percentage range, or ratio range recited herein are to be understood to include concentrations, percentages or ratios of any integer within that range and fractions thereof, such as one tenth and one hundredth of an integer, unless otherwise indicated.
• any number range recited herein relating to any physical feature, such as polymer subunits, size or thickness, are to be understood to include any integer within the recited range, unless otherwise indicated.
• the term “about” refers to + 15% of any indicated structure, value, or range.
• “A” and “an” refer to one or more of the indicated items.
• a polymer refers to both one polymer or a mixture comprising two or more polymers
• a multifunctional compound refers not only to a single multifunctional compound but also to a combination of two or more of the same or different multifunctional compounds
• a reactive group refers to a combination of reactive groups as well as to a single reactive group, and the like.
• the present invention provides polymeric compositions which greatly increase the ability to inhibit the formation of reactive scar tissue on, or around, the surface of a device or implant or at a treatment site.
• Numerous polymeric compositions and therapeutic agents are described herein.
• the present invention provides for the combination of compositions (e.g., polymers) which include one or more therapeutic agents, described below. Also described in more detail below are methods for making and methods for utilizing such compositions.
• the present invention discloses pharmaceutical agents which inhibit one or more aspects of the production of excessive fibrous (scar) tissue.
• Suitable fibrosis-inhibiting or stenosis-inhibiting agents may be readily determined based upon the in vitro and in vivo (animal) models such as those provided in Examples 20-33.
• Agents which inhibit fibrosis may be identified through in vivo models including inhibition of intimal hyperplasia development in the rat balloon carotid artery model (Examples 25 and 33).
• the assays set forth in Examples 24 and 32 may be used to determine whether an agent is able to inhibit cell proliferation in fibroblasts and/or smooth muscle cells.
• the agent has an IC 50 for inhibition of cell proliferation within a range of about 10 "6 to about 10 "10 M.
• the assay set forth in Example 28 may be used to determine whether an agent may inhibit migration of fibroblasts and/or smooth muscle cells.
• the agent has an IC 50 for inhibition of cell migration within a range of about 10 "6 to about 10 "9 M.
• Assays set forth herein may be used to determine whether an agent is able to inhibit inflammatory processes, including nitric oxide production in macrophages (Example 20), and/or TNF-alpha production by macrophages (Example 21 ), and/or IL-1 beta production by macrophages (Example 29), and/or IL-8 production by macrophages (Example 30), and/or inhibition of MCP- 1 by macrophages (Example 31).
• the agent has an IC 50 for inhibition of any one of these inflammatory processes within a range of about 10 " ⁇ to about 10 "10 M.
• the assay set forth in Example 26 may be used to determine whether an agent is able to inhibit MMP production.
• the agent has an IC 50 for inhibition of MMP production within a range of about 10 "4 to about 10 "8 M.
• the assay set forth in Example 27 (also known as the CAM assay) may be used to determine whether an agent is able to inhibit angiogenesis.
• the agent has an IC 50 for inhibition of angiogenesis within a range of about 10 "6 to about 10 "10 M.
• Agents which reduce the formation of surgical adhesions may be identified through in vivo models including the rabbit surgical adhesions model (Examples 23, 42 and 43) and the rat caecal sidewall model (Example 22).
• the pharmacologically active fibrosis- inhibiting compound is an angiogenesis inhibitor (e.g., 2-ME (NSC-659853), Pl- 88 (D-mannose, 0-6-0-phosphono-alpha-D-mannopyranosyl-(1 -3)-0-alpha-D- mannopyranosyl-(1-3)-0-alpha-D-mannopyranosyl-(1-3)-0-alpha-D- mannopyranosyl-(1-2)- hydrogen sulfate), thalidomide (1 H-isoindole-1 ,3(2H)- dione, 2-(2,6-dioxo-3
• the pharmacologically active fibrosis- inhibiting compound is a 5-lipoxygenase inhibitor or antagonist (e.g., Wy-50295 (2-naphthaleneacetic acid, alpha-methyl-6-(2-quinolinylmethoxy)-, (S)-), ONO- LP-269 (2,11 ,14-eicosatrienamide, N-(4-hydroxy-2-(1 H-tetrazol-5-yl)-8- quinolinyl)-, (E,Z,Z)-), licofelone (1 H-pyrrolizine-5-acetic acid, 6-(4- chlorophenyl)-2,3-dihydro-2,2-dimethyl-7-phenyl-), CMI-568 (urea, N-butyl-N- hydroxy-N'-(4-(3-(methylsulfonyl)-2-propoxy-5-(tetrahydro-5
• the pharmacologically active fibrosis- inhibiting compound is a chemokine receptor antagonist which inhibits one or more subtypes of CCR (1 , 3, and 5) (e.g., ONO-4128 (1 ,4,9- triazaspiro(5.5)undecane-2,5-dione, 1-butyl-3-(cyclohexylmethyl)-9-((2,3- dihydro-1 ,4-benzodioxin-6-yl)methyl-), L-381 , CT-112 (L-arginine, L-threonyl-L- threonyl-L-seryl-L-glutaminyl-L-valyl-L-arginyl-L-prolyl-), AS-900004, SCH-C, ZK-811752, PD-172084, UK-427857, SB-380732, vMIP II, SB-265
• chemokine receptor antagonists include a-lmmunokine-NNS03, BX-471 , CCX-282, Sch-350634; Sch-351125; Sch- 417690; SCH-C, and analogues and derivatives thereof.
• the pharmacologically active fibrosis- inhibiting compound is a cell cycle inhibitor.
• Representative examples of such agents include taxanes (e.g., paclitaxel (discussed in more detail below) and docetaxel) (Schiff et al., Nature 277:665-667, 1979; Long and Fairchild, Cancer Research 54:4355-4361 , 1994; Ringel and Horwitz, J. Nat'l Cancer Inst. 83(4):288-291 , 1991 ; Pazdur et al., Cancer Treat. Rev. 79(40):351-386, 1993), etanidazole, nimorazole (B.A. Chabner and D.L. Longo.
• Nitroimidazole derivative, production thereof, and radiosensitizer containing the same as active ingredient U.S. Patent No. 5,270,330, Dec 14, 1993; T. Suzuki. 2-Nitroimidazole derivative, production thereof and radiosensitizer containing the same as active ingredient; Patent EP 0 513 351 B1 , Jan. 24, 1991), fluorine-containing nitroazole derivatives (T. Kagiya. Fluorine-containing nitroazole derivatives and radiosensitizer comprising the same.
• copper M.J. Abrams. Copper Radiosensitizers.
• Heterocyclic compound derivative, its production and radiosensitizer containing said derivative as active ingredient Publication Number 63170375 A (Japan), Jan. 7, 1987), fluorine containing 3-nitro-1 ,2,4- triazole (T. Kagitani et al. Novel fluorine-containing 3-nitro-1 ,2,4-triazole and radiosensitizer containing same compound.
• Radiosensitizer for Hypoxic cell Publication Number 61010511 A (Japan), Jun. 26, 1984), Nitrothiazole (T. Kagitani et al. Radiation-sensitizing agent. Publication Number 61167616 A (Japan) Jan. 22, 1985), imidazole derivatives (S. Inayma et al. Imidazole derivative. Publication Number 6203767 A (Japan) Aug. 1 ,1985; Publication Number 62030768 A (Japan) Aug. 1 , 1985;
• camptothecin Ewend M.G. et al. Local delivery of chemotherapy and concurrent external beam radiotherapy prolongs survival in metastatic brain tumor models. Cancer Research 56(22):5217-5223, 1996) and paclitaxel (Tishler R.B. et al. Taxol: a novel radiation sensitizer. International Journal of Radiation Oncology and Biological Physics 22(3):613- 617, 1992).
• a number of the above-mentioned cell cycle inhibitors also have a wide variety of analogues and derivatives, including, but not limited to, cisplatin, cyclophosphamide, misonidazole, tiripazamine, nitrosourea, mercaptopurine, methotrexate, flurouracil, epirubicin, doxorubicin, vindesine and etoposide.
• Analogues and derivatives include (CPA) 2 Pt(DOLYM) and (DACH)Pt(DOLYM) cisplatin (Choi et al., Arch. Pharmacal Res. 22(2):151-156, 1999), Cis-
• gem-diphosphonate cisplatin analogues (FR 2683529), (meso-1 ,2-bis(2,6-dichloro-4-hydroxyplenyl)ethylenediamine) dichloroplatinum(ll) (Bednarski et al., J. Med. Chem. 35(23):4479-85, 1992), cisplatin analogues containing a tethered dansyl group (Hartwig et al., J. Am. Chem. Soc. 774(21):8292-3, 1992), platinum(ll) polyamines (Siegmann et al., Inorg. Met.-Containing Polym.
• 6-mercaptopurine (6-MP) (Kashida et al., Biol. Pharm. Bull. 78(11 ):1492-7, 1995), 7,8-polymethyleneimidazo-1 ,3,2- diazaphosphorines (Nilov et al., Mendeleev Commun. 2:67, 1995), azathioprine (Chifotides et al., J. Inorg. Biochem. 56(4):249-64, 1994), methyl-D- glucopyranoside mercaptopurine derivatives (Da Suva et al., Eivr. J. Med.
• methotrexate tetrahydroquinazoline analogue (Gangjee, et al., J. Heterocycl. Chem. 32(1):243-8, 1995), N-( ⁇ -aminoacyl) methotrexate derivatives (Cheung et al., Pteridines 3(1-2):101-2, 1992), biotin methotrexate derivatives (Fan et al., Pteridines 3(1-2):131-2, 1992), D-glutamic acid or D-erythrou, threo-4- fluoroglutamic acid methotrexate analogues (McGuire et al., Biochem. Pharmacol.
• Pteridines Folic Acid Deriv., 1154-7, 1989 N-(L- ⁇ -aminoacyl) methotrexate derivatives (Cheung et al., Heterocycles 28(2):751-8, 1989), meta and ortho isomers of aminopterin (Rosowsky et al., J. Med. Chem. 32(12):2582, 1989), hydroxymethylmethotrexate (DE 267495), ⁇ -fluoromethotrexate (McGuire et al., Cancer Res. 49(16):4517-25, 1989), polyglutamyl methotrexate derivatives (Kumar et al., Cancer Res.
• the cell cycle inhibitor is paclitaxel, a compound which disrupts mitosis (M-phase) by binding to tubulin to form abnormal mitotic spindles or an analogue or derivative thereof.
• paclitaxel is a highly derivatized diterpenoid (Wani et al., J. Am. Chem. Soc.
• Taxus brevifolia Pacific Yew
• Taxomyces Andreanae and Endophytic Fungus of the Pacific Yew Stierle et al., Science 60:214-216, 1993.
• “Paclitaxel” (which should be understood herein to include formulations, prodrugs, analogues and derivatives such as, for example, TAXOL (Bristol Myers Squibb, New York, NY, TAXOTERE (Aventis Pharmaceuticals, France), docetaxel, 10-desacetyl analogues of paclitaxel and 3'N-desbenzoyl-3'N-t- butoxy carbonyl analogues of paclitaxel) may be readily prepared utilizing techniques known to those skilled in the art (see, e.g., Schiff et al., Nature 277:665-667, 1979; Long and Fairchild, Cancer Research 54:4355-4361 , 1994; Ringel and Horwitz, J.
• paclitaxel derivatives or analogues include 7-deoxy-docetaxol, 7,8-cyclopropataxanes, N-substituted 2-azetidones, 6,7-epoxy paclitaxels, 6,7-modified paclitaxels, 10-desacetoxytaxol, 10- deacetyltaxol (from 10-deacetylbaccatin III), phosphonooxy and carbonate derivatives of taxol, taxol 2',7-di(sodium 1 ,2-benzenedicarboxylate, 10- desacetoxy-11 ,12-dihydrotaxol-10,12(18)-diene derivatives, 10- desacetoxytaxol, Protaxol (2'-and/or 7-O-ester derivatives), (2'-and/or 7-0- carbonate derivatives), asymmetric synthesis of taxol side chain, fluoro taxols, 9-deoxotaxane, (13-acetyl
• gray-highlighted portions may be substituted and the non-highlighted portion is the taxane core.
• a side-chain (labeled "A" in the diagram) is desirably present in order for the compound to have good activity as a cell cycle inhibitor.
• Examples of compounds having this structure include paclitaxel (Merck Index entry 7117), docetaxol (TAXOTERE, Merck Index entry 3458), and 3'- desphenyl-3'-(4-ntirophenyl)-N-debenzoyl-N-(t-butoxycarbonyl)-10- deacetyltaxol.
• suitable taxanes such as paclitaxel and its analogues and derivatives are disclosed in U.S. Patent No. 5,440,056 as having the structure (C2):
• X may be oxygen (paclitaxel), hydrogen (9-deoxy derivatives), thioacyl, or dihydroxyl precursors;
• R-i is selected from paclitaxel or TAXOTERE side chains or alkanoyl of the formula (C3)
• R 7 is selected from hydrogen, alkyl, phenyl, alkoxy, amino, phenoxy (substituted or unsubstituted);
• Rs is selected from hydrogen, alkyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, phenyl (substituted or unsubstituted), alpha or beta- naphthyl;
• Rg is selected from hydrogen, alkanoyl, substituted alkanoyl, and aminoalkanoyl; where substitutions refer to hydroxyl, sulfhydryl, allalkoxyl, carboxyl, halogen, thioalkoxyl, N,N-dimethylamino, alkylamino, dialkylamino, nitro, and -OS0 3 H, and/or may refer to groups containing such substitutions;
• R 2 is selected from hydrogen or oxygen-containing groups, such as hydrogen, hydroxyl, alkoyl, alkanoyloxy, aminoalkano
• the paclitaxel analogues and derivatives useful as cell cycle inhibitors are disclosed in PCT International Patent Application No. WO 93/10076.
• the analogue or derivative should have a side chain attached to the taxane nucleus at C 3 , as shown in the structure below (formula C4), in order to confer antitumor activity to the taxane.
• WO 93/10076 discloses that the taxane nucleus may be substituted at any position with the exception of the existing methyl groups.
• the substitutions may include, for example, hydrogen, alkanoyloxy, alkenoyloxy, aryloyloxy.
• oxo groups may be attached to carbons labeled 2, 4, 9, and/or 10.
• an oxetane ring may be attached at carbons 4 and 5.
• an oxirane ring may be attached to the carbon labeled 4.
• the taxane-based cell cycle inhibitor useful in the present invention is disclosed in U.S. Patent 5,440,056, which discloses 9- deoxo taxanes.
• the taxane ring may be substituted at the carbons labeled 1 , 7 and 10 (independently) with H, OH, O-R, or O-CO-R where R is an alkyl or an aminoalkyl. As well, it may be substituted at carbons labeled 2 and 4 (independently) with aryol, alkanoyl, aminoalkanoyl or alkyl groups.
• the side chain of formula (C3) may be substituted at R 7 and R 8 (independently) with phenyl rings, substituted phenyl rings, linear alkanes/alkenes, and groups containing H, O or N.
• R 9 may be substituted with H, or a substituted or unsubstituted alkanoyl group.
• Taxanes in general, and paclitaxel is particular, is considered to function as a cell cycle inhibitor by acting as an anti-microtubule agent, and more specifically as a stabilizer. These compounds have been shown useful in the treatment of proliferative disorders, including: non-small cell (NSC) lung; small cell lung; breast; prostate; cervical; endometrial; head and neck cancers.
• NSC non-small cell
• the anti-microtuble agent is albendazole (carbamic acid, (5-(propylthio)-1 H-benzimidazol-2-yl)-, methyl ester), LY-355703 (1 ,4-dioxa-8,11-diazacyclohexadec-13-ene-2,5,9,12-tetrone, 10-((3-chloro-4-methoxyphenyl)methyl)-6,6-dimethyl-3-(2-methylpropyl)-16- ((1S)-1-((2S,3R)-3-phenyloxiranyl)ethyl)-, (3S,10R,13E,16S)-), vindesine (vincaleukoblastine, 3-(aminocarbonyl)-04-deacetyl-3-de(methoxycarbonyl)-), or WAY-174286
• the cell cycle inhibitor is a vinca alkaloid. Vinca alkal
• R-t can be a formyl or methyl group or alternately H.
• R-i can also be an alkyl group or an aldehyde-substituted alkyl (e.g., CH 2 CHO).
• R 2 is typically a CH 3 or NH 2 group. However it can be alternately substituted with a lower alkyl ester or the ester linking to the dihydroindole core may be substituted with C(0)-R where R is NH 2 , an amino acid ester or a peptide ester.
• R 3 is typically C(0)CH 3 , CH 3 or H.
• a protein fragment may be linked by a bifunctional group, such as maleoyl amino acid.
• R 3 can also be substituted to form an alkyl ester which may be further substituted.
• R may be -CH 2 - or a single bond.
• R 5 and R 6 may be H, OH or a lower alkyl, typically -CH 2 CH 3 .
• R 6 and R 7 may together form an oxetane ring.
• R 7 may alternately be H.
• Further substitutions include molecules wherein methyl groups are substituted with other alkyl groups, and whereby unsaturated rings may be derivatized by the addition of a side group such as an alkane, alkene, alkyne, halogen, ester, amide or amino group.
• Exemplary vinca alkaloids are vinblastine, vincristine, vincristine sulfate, vindesine, and vinorelbine, having the structures:
• the cell cycle inhibitor is a camptothecin, or an analog or derivative thereof.
• Camptothecins have the following general structure.
• X is typically O, but can be other groups, e.g., NH in the case of 21-lactam derivatives.
• R-i is typically H or OH, but may be other groups, e.g., a terminally hydroxylated C -3 alkane.
• R 2 is typically H or an amino containing group such as (CH 3 ) 2 NHCH 2 , but may be other groups e.g., N0 2 , NH 2 , halogen (as disclosed in, e.g., U.S. Patent 5,552,156) or a short alkane containing these groups.
• R 3 is typically H or a short alkyl such as C 2 H 5 .
• R is typically H but may be other groups, e.g., a methylenedioxy group with R-i.
• Exemplary camptothecin compounds include topotecan, irinotecan (CPT-11 ), 9-aminocamptothecin, 21 -lactam-20(S)-camptothecin, 10,11-methylenedioxycamptothecin, SN-38, 9-nitrocamptothecin, 10- hydroxycamptothecin.
• Exemplary compounds have the structures:
• Camptothecins have the five rings shown here.
• the ring labeled E must be intact (the lactone rather than carboxylate form) for maximum activity and minimum toxicity.
• These compounds are useful to as cell cycle inhibitors, where they can function as topoisomerase I inhibitors and/or DNA cleavage agents. They have been shown useful in the treatment of proliferative disorders, including, for example, NSC lung; small cell lung; and cervical cancers.
• the cell cycle inhibitor is a podophyllotoxin, or a derivative or an analogue thereof.
• Exemplary compounds of this type are etoposide orteniposide, which have the following structures:
• These compounds are thought to function as cell cycle inhibitors by being topoisomerase II inhibitors and/or by DNA cleaving agents. They have been shown useful as antiproliferative agents in, e.g., small cell lung, prostate, and brain cancers, and in retinoblastoma.
• DNA topoisomerase inhibitor is lurtotecan dihydrochloride (11 H-1 ,4-dioxino(2,3-g)pyrano(3',4':6,7)indolizino(1 ,2- b)quinoline-9,12(8H,14H)-dione, 8-ethyl-2,3-dihydro-8-hydroxy-15-((4-methyl-1- piperazinyl)methyl)-, dihydrochloride, (S)-).
• the cell cycle inhibitor is an anthracycline.
• Anthracyclines have the following general structure, where the R groups may be a variety of organic groups:
• R-i is CH 3 or CH 2 OH
• R 2 is daunosamine or H
• R 3 and R 4 are independently one of OH, N0 2 , NH 2 , F, CI, Br, I, CN, H or groups derived from these
• R 5 - 7 are all H or
• R 5 and R 6 are H and R 7 and Rs are alkyl or halogen, or vice versa
• R 7 and R 8 are H and R 5 and Re are alkyl or halogen.
• R 2 may be a conjugated peptide.
• R 5 may be OH or an ether linked alkyl group.
• R- t may also be linked to the anthracycline ring by a group other than C(O), such as an alkyl or branched alkyl group having the C(O) linking moiety at its end, such as -CH 2 CH(CH 2 -X)C(0)-R 1 > wherein X is H or an alkyl group (see, e.g., U.S. Patent 4,215,062).
• R 3 may have the following structure:
• R 9 is OH either in or out of the plane of the ring, or is a second sugar moiety such as R 3 .
• R 10 may be H or form a secondary amine with a group such as an aromatic group, saturated or partially saturated 5 or 6 membered heterocyclic having at least one ring nitrogen (see U.S. Patent 5,843,903). Alternately, R 10 may be derived from an amino acid, having the structure -
• C(0)CH(NHRn)(R 12 ), in which Rn is H, or forms a C 3-4 membered alkylene with Ri2- R12 may be H, alkyl, aminoalkyl, amino, hydroxy, mercapto, phenyl, benzyl or methylthio (see U.S. Patent 4,296,105).
• exemplary anthracyclines are doxorubicin, daunorubicin, idarubicin, epirubicin, pirarubicin, zorubicin, and carubicin. Suitable compounds have the structures:
• anthracyclines are anthramycin, mitoxantrone, menogaril, nogalamycin, aclacinomycin A, olivomycin A, chromomycin A 3 , and plicamycin having the structures:
• the cell cycle inhibitor is a platinum compound.
• suitable platinum complexes may be of Pt(ll) or Pt(IV) and have this basic structure:
• Pt(ll) complexes Z ⁇ and Z 2 are non-existent.
• Pt(IV) Z-i and Z may be anionic groups such as halogen, hydroxy, carboxylate, ester, sulfate or phosphate. See, e.g., U.S. Patent Nos. 4,588,831 and 4,250,189.
• Suitable platinum complexes may contain multiple Pt atoms. See, e.g., U.S. Patent Nos. 5,409,915 and 5,380,897.
• platinum compounds are cisplatin, carboplatin, oxaliplatin, and miboplatin having the structures:
• Oxaliplatin These compounds are thought to function as cell cycle inhibitors by binding to DNA, i.e., acting as alkylating agents of DNA. These compounds have been shown useful in the treatment of cell proliferative disorders, including, e.g., NSC lung; small cell lung; breast; cervical; brain; head and neck; esophageal; retinoblastom; liver; bile duct; bladder; penile; and vulvar cancers; and soft tissue sarcoma.
• the cell cycle inhibitor is a nitrosourea. Nitrosourease have the following general structure (C5), where typical R groups are shown below.
• R groups include cyclic alkanes, alkanes, halogen substituted groups, sugars, aryl and heteroaryl groups, phosphonyl and sulfonyl groups.
• R may suitably be CH 2 - C(X)(Y)(Z), wherein X and Y may be the same or different members of the following groups: phenyl, cyclyhexyl, or a phenyl or cyclohexyl group substituted with groups such as halogen, lower alkyl (C ⁇ -4 ), trifluore methyl, cyano, phenyl, cyclohexyl, lower alkyloxy (C -4 ).
• Z has the following structure: -alkylene-N-R ⁇ R 2 , where R-i and R 2 may be the same or different members of the following group: lower alkyl (C ⁇ -4 ) and benzyl, or together Ri and R 2 may form a saturated 5 or 6 membered heterocyclic such as pyrrolidine, piperidine, morfoline, thiomorfoline, N-lower alkyl piperazine, where the heterocyclic may be optionally substituted with lower alkyl groups.
• R and R' of formula (C5) may be the same or different, where each may be a substituted or unsubstituted hydrocarbon having 1-10 carbons.
• Substitutions may include hydrocarbyl, halo, ester, amide, carboxylic acid, ether, thioether and alcohol groups.
• R of formula (C5) may be an amide bond and a pyranose structure (e.g., methyl 2'-(N-(N-(2-chloroethyl)- N-nitroso-carbamoyl)-glycyl)amino-2'-deoxy- ⁇ -D-glucopyranoside).
• a pyranose structure e.g., methyl 2'-(N-(N-(2-chloroethyl)- N-nitroso-carbamoyl)-glycyl
• R of formula (C5) may be an alkyl group of 2 to 6 carbons and may be substituted with an ester, sulfonyl, or hydroxyl group. It may also be substituted with a carboxylic acid or CONH 2 group.
• exemplary nitrosoureas are BCNU (carmustine), methyl-CCNU (semustine), CCNU (lomustine), ranimustine, nimustine, chlorozotocin, fotemustine, and streptozocin, having the structures:
• the cell cycle inhibitor is a nitroimidazole, where exemplary nitroimidazoles are metronidazole, benznidazole, etanidazole, and misonidazole, having the structures:
• the cell cycle inhibitor is a folic acid antagonist, such as methotrexate or derivatives or analogues thereof, including edatrexate, trimetrexate, raltitrexed, piritrexim, denopterin, tomudex, and pteropterin.
• Methotrexate analogues have the following general structure:
• R group may be selected from organic groups, particularly those groups set forth in U.S. Patent Nos. 5,166,149 and 5,382,582.
• Ri may be N
• R 2 may be N or C(CH 3 )
• R 3 and R 3 ' may H or alkyl, e.g., CH 3
• R 4 may be a single bond or NR, where R is H or alkyl group.
• R 5 , 6,8 may be H, OCH 3 , or alternately they can be halogens or hydro groups.
• R 7 is a side chain of the general structure:
• the carboxyl groups in the side chain may be esterified or form a salt such as a Zn + salt.
• R 9 and R- ⁇ 0 can be NH 2 or may be alkyl substituted.
• Exemplary folic acid antagonist compounds have the structures:
• the cell cycle inhibitor is a cytidine analogue, such as cytarabine or derivatives or analogues thereof, including enocitabine, FMdC ((E(-2'-deoxy-2'-(fluoromethylene)cytidine), gemcitabine, 5-azacitidine, ancitabine, and 6-azauridine.
• cytidine analogue such as cytarabine or derivatives or analogues thereof, including enocitabine, FMdC ((E(-2'-deoxy-2'-(fluoromethylene)cytidine), gemcitabine, 5-azacitidine, ancitabine, and 6-azauridine.
• Exemplary compounds have the structures:
• the cell cycle inhibitor is a pyrimidine analogue.
• the pyrimidine analogues have the general structure: wherein positions 2', 3' and 5' on the sugar ring (R 2 , R 3 and R , respectively) can be H, hydroxyl, phosphoryl (see, e.g., U.S. Patent 4,086,417) or ester (see, e.g., U.S. Patent 3,894,000).
• Esters can be of alkyl, cycloalkyl, aryl or heterocyclo/aryl types.
• the 2' carbon can be hydroxylated at either R 2 or R 2 ', the other group is H.
• the 2' carbon can be substituted with halogens e.g., fluoro or difluoro cytidines such as Gemcytabine.
• the sugar can be substituted for another heterocyclic group such as a furyl group or for an alkane, an alkyl ether or an amide linked alkane such as C(0)NH(CH 2 ) 5 CH 3 .
• the 2° amine can be substituted with an aliphatic acyl (Ri) linked with an amide (see, e.g., U.S. Patent 3,991 ,045) or urethane (see, e.g., U.S. Patent 3,894,000) bond. It can also be further substituted to form a quaternary ammonium salt.
• R 5 in the pyrimidine ring may be N or CR, where R is H, halogen containing groups, or alkyl (see, e.g., U.S. Patent No. 4,086,417).
• R 8 is H or R 7 and R 8 together can form a double bond or Rs can be X, where X is:
• U.S. Patent No. 3,894,000 see, e.g., 2'-0-palmityl-ara-cytidine, 3'-0-benzoyl-ara-cytidine, and more than 10 other examples
• U.S. Patent No. 3,991 ,045 see, e.g., N4-acyl-1- ⁇ -D-arabinofuranosylcytosine, and numerous acyl groups derivatives as listed therein, such as palmitoyl.
• the cell cycle inhibitor is a fluoropyrimidine analogue, such as 5-fluorouracil, or an analogue or derivative thereof, including carmofur, doxifluridine, emitefur, tegafur, and floxuridine.
• fluoropyrimidine analogue such as 5-fluorouracil
• an analogue or derivative thereof including carmofur, doxifluridine, emitefur, tegafur, and floxuridine.
• Exemplary compounds have the structures:
• fluoropyrimidine analogues include 5-FudR (5- fluoro-deoxyuridine), or an analogue or derivative thereof, including 5- iododeoxyuridine (5-ludR), 5-bromodeoxyuridine (5-BudR), fluorouridine triphosphate (5-FUTP), and fluorodeoxyuridine monophosphate (5-dFUMP).
• 5-FudR 5- fluoro-deoxyuridine
• an analogue or derivative thereof including 5- iododeoxyuridine (5-ludR), 5-bromodeoxyuridine (5-BudR), fluorouridine triphosphate (5-FUTP), and fluorodeoxyuridine monophosphate (5-dFUMP).
• Exemplary compounds have the structures:
• the cell cycle inhibitor is a purine analogue.
• Purine analogues have the following general structure.
• N signifies nitrogen and V, W, X, Z can be either carbon or nitrogen with the following provisos.
• Ring A may have 0 to 3 nitrogen atoms in its structure. If two nitrogens are present in ring A, one must be in the W position. If only one is present, it must not be in the Q position. V and Q must not be simultaneously nitrogen. Z and Q must not be simultaneously nitrogen. If Z is nitrogen, R 3 is not present.
• R ⁇ -3 are independently one of H, halogen, C ⁇ -7 alkyl, C ⁇ -7 alkenyl, hydroxyl, mercapto, C ⁇ -7 alkylthio, C ⁇ -7 alkoxy, C - alkenyloxy, aryl oxy, nitro, primary, secondary or tertiary amine containing group.
• R 5-8 are H or up to two of the positions may contain independently one of OH, halogen, cyano, azido, substituted amino, R 5 and R can together form a double bond.
• Y is H, a C-i_ 7 alkylcarbonyl, or a mono- di or tri phosphate.
• Exemplary suitable purine analogues include 6-mercaptopurine, thiguanosine, thiamiprine, cladribine, fludaribine, tubercidin, puromycin, pentoxyfilline; where these compounds may optionally be phosphorylated.
• Exemplary compounds have the structures: Pentoxyfilline
• the cell cycle inhibitor is a nitrogen mustard.
• suitable nitrogen mustards are known and are suitably used as a cell cycle inhibitor in the present invention.
• Suitable nitrogen mustards are also known as cyclophosphamides.
• a preferred nitrogen mustard has the general structure:
• alkane typically CH 2 CH(CH 3 )CI, or a polycyclic group such as B, or a substituted phenyl such as C or a heterocyclic group such as D.
• Patent No. 3,808,297 wherein A is:
• R ⁇ -2 are H or CH 2 CH 2 CI;
• R 3 is H or oxygen-containing groups such as hydroperoxy; and
• R 4 can be alkyl, aryl, heterocyclic.
• the cyclic moiety need not be intact. See, e.g., U.S. Patent Nos. 5,472,956, 4,908,356, 4,841 ,085 that describe the following type of structure:
• R 1 is H or CH 2 CH 2 CI
• R 2-6 are various substituent groups.
• exemplary nitrogen mustards include methylchloroethamine, and analogues or derivatives thereof, including methylchloroethamine oxide hydrohchloride, novembichin, and mannomustine (a halogenated sugar).
• Exemplary compounds have the structures:
• the nitrogen mustard may be cyclophosphamide, ifosfamide, perfosfamide, or torofosfamide, where these compounds have the structures: R, Cyclophosphamid e H CH 2 CH 2 CI H Ifosfamide CH 2 CH 2 CI H H Perfosfamide CH 2 CH 2 CI H OOH Torofosfamide CH 2 CH 2 CI CH 2 CH 2 CI H
• the nitrogen mustard may be estramustine, or an analogue or derivative thereof, including phenesterine, prednimustine, and estramustine P0 4 .
• suitable nitrogen mustard type cell cycle inhibitors of the present invention have the structures:
• the nitrogen mustard may be chlorambucil, or an analogue or derivative thereof, including melphalan and chlormaphazine.
• suitable nitrogen mustard type cell cycle inhibitors of the present invention have the structures:
• the nitrogen mustard may be uracil mustard, which has the structure:
• the nitrogen mustards are thought to function as cell cycle inhibitors by serving as alkylating agents for DNA.
• Nitrogen mustards have been shown useful in the treatment of cell proliferative disorders including, for example, small cell lung, breast, cervical, head and neck, prostate, retinoblastoma, and soft tissue sarcoma.
• the cell cycle inhibitor of the present invention may be a hydroxyurea. Hydroxyureas have the following general structure:
• Suitable hydroxyureas are disclosed in, for example, U.S. Patent No. 6,080,874, wherein Ri is:
• R 2 is an alkyl group having 1-4 carbons and R 3 is one of H, acyl, methyl, ethyl, and mixtures thereof, such as a methylether.
• R-i is a cycloalkenyl group, for example N-(3-(5-(4- fluorophenylthio)-furyl)-2-cyclopenten-1-yl)N-hydroxyurea
• R 2 is H or an alkyl group having 1 to 4 carbons and R 3 is H
• X is H or a cation.
• Other suitable hydroxyureas are disclosed in, e.g., U.S.
• R-i is a phenyl group substituted with on or more fluorine atoms
• R 2 is a cyclopropyl group
• R 3 and X is H.
• Other suitable hydroxyureas are disclosed in, e.g., U.S. Patent No. 4,299,778, wherein R-i is a phenyl group substituted with on or more fluorine atoms; R 2 is a cyclopropyl group; and R 3 and X is H.
• Other suitable hydroxyureas are disclosed in, e.g., U.S. Patent
• hydroxy urea has the structure:
• Hydroxyureas are thought to function as cell cycle inhibitors by serving to inhibit DNA synthesis.
• the cell cycle inhibitor is a mytomicin, such as mitomycin C, or an analogue or derivative thereof, such as porphyromycin.
• Exemplary compounds have the structures: R Mitomycin C H Porphyromycin CH 3 (N-methyl Mitomycin C)
• the cell cycle inhibitor is an alkyl sulfonate, such as busulfan, or an analogue or derivative thereof, such as treosulfan, improsulfan, piposulfan, and pipobroman.
• alkyl sulfonate such as busulfan
• an analogue or derivative thereof such as treosulfan, improsulfan, piposulfan, and pipobroman.
• Exemplary compounds have the structures: •
• the cell cycle inhibitor is a benzamide.
• the cell cycle inhibitor is a nicotinamide.
• X is either O or S; A is commonly NH 2 or it can be OH or an alkoxy group; B is N or C-R 4 , where R is H or an ether-linked hydroxylated alkane such as OCH 2 CH 2 OH, the alkane may be linear or branched and may contain one or more hydroxyl groups. Alternately, B may be N-R 5 in which case the double bond in the ring involving B is a single bond. R 5 may be H, and alkyl or an aryl group (see, e.g., U.S. Patent No.
• R 2 is H, OR 6 , SR 6 or NHR 6 , where R 6 is an alkyl group; and R 3 is H, a lower alkyl, an ether linked lower alkyl such as -O-Me or-O-ethyl (see, e.g., U.S. Patent No. 5,215,738).
• Suitable benzamide compounds have the structures:
• the cell cycle inhibitor is a halogenated sugar, such as mitolactol, or an analogue or derivative thereof, including mitobronitol and mannomustine.
• Examplary compounds have the structures:
• the cell cycle inhibitor is a diazo compound, such as azaserine, or an analogue or derivative thereof, including 6-diazo-5- oxo-L-norleucine and 5-diazouracil (also a pyrimidine analog).
• Examplary compounds have the structures: R 1 R 2 Azaserine O single bond 6-diazo-5-oxo- L-norleucine single bond CH.
• pazelliptine wortmannin; metoclopramide; RSU; buthionine sulfoxime; tumeric; curcumin; AG337, a thymidylate synthase inhibitor; levamisole; lentinan, a polysaccharide; razoxane, an EDTA analogue; indomethacin; chlorpromazine; ⁇ and ⁇ interferon; MnBOPP; gadolinium texaphyrin; 4-amino-1 ,8-naphthalimide; staurosporine derivative of CGP; and SR-2508.
• the cell cycle inhibitor is a DNA alylating agent.
• the cell cycle inhibitor is an anti-microtubule agent.
• the cell cycle inhibitor is a topoisomerase inhibitor.
• the cell cycle inhibitor is a DNA cleaving agent.
• the cell cycle inhibitor is an antimetabolite.
• the cell cycle inhibitor functions by inhibiting adenosine deaminase (e.g., as a purine analogue).
• the cell cycle inhibitor functions by inhibiting purine ring synthesis and/or as a nucleotide interconversion inhibitor (e.g., as a purine analogue such as mercaptopurine).
• the cell cycle inhibitor functions by inhibiting dihydrofolate reduction and/or as a thymidine monophosphate block (e.g., methotrexate). In another aspect, the cell cycle inhibitor functions by causing DNA damage (e.g., bleomycin).
• a thymidine monophosphate block e.g., methotrexate
• the cell cycle inhibitor functions by causing DNA damage (e.g., bleomycin).
• the cell cycle inhibitor functions as a DNA intercalation agent and/or RNA synthesis inhibition (e.g., doxorubicin, aclarubicin, or detorubicin (acetic acid, diethoxy-, 2-(4-((3-amino-2,3,6-trideoxy-alpha-L-lyxo-hexopyranosyl)oxy)- 1 ,2,3,4,6,11-hexahydro-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-2- naphthacenyl)-2-oxoethyl ester, (2S-cis)-)).
• doxorubicin e.g., doxorubicin, aclarubicin, or detorubicin (acetic acid, diethoxy-, 2-(4-((3-amino-2,3,6-trideoxy-alpha-L-lyxo-hexopyranosyl)oxy)- 1 ,2,3,
• the cell cycle inhibitor functions by inhibiting pyrimidine synthesis (e.g., N-phosphonoacetyl-L- aspartate). In another aspect, the cell cycle inhibitor functions by inhibiting ribonucleotides (e.g., hydroxyurea). In another aspect, the cell cycle inhibitor functions by inhibiting thymidine monophosphate (e.g., 5-fluorouracil). In another aspect, the cell cycle inhibitor functions by inhibiting DNA synthesis (e.g., cytarabine). In another aspect, the cell cycle inhibitor functions by causing DNA adduct formation (e.g., platinum compounds). In another aspect, the cell cycle inhibitor functions by inhibiting protein synthesis (e.g., L- asparginase).
• pyrimidine synthesis e.g., N-phosphonoacetyl-L- aspartate
• the cell cycle inhibitor functions by inhibiting ribonucleotides (e.g., hydroxyurea).
• the cell cycle inhibitor functions by inhibiting th
• the cell cycle inhibitor functions by inhibiting microtubule function (e.g., taxanes).
• the cell cycle inhibitor acts at one or more of the steps in the biological pathway shown in FIG. 1. Additional cell cycle inhibitor s useful in the present invention, as well as a discussion of the mechanisms of action, may be found in Hardman J.G., Limbird L.E. Molinoff R.B., Ruddon R W, Gilman A.G. editors, Chemotherapy of Neoplastic Diseases in Goodman and Gilman's The Pharmacological Basis of Therapeutics Ninth Edition, McGraw-Hill Health Professions Division, New York, 1996, pages 1225-1287. See also U.S. Patent Nos.
• the cell-cycle inhibitor is camptothecin, mitoxantrone, etoposide, 5-fluorouracil, doxorubicin, methotrexate, peloruside A, mitomycin C, or a CDK-2 inhibitor or an analogue or derivative of any member of the class of listed compounds.
• the cell-cycle inhibitor is HTI-286, plicamycin; or mithramycin, or an analogue or derivative thereof.
• cell cycle inhibitors also include, e.g., 7- hexanoyltaxol (QP-2), cytochalasin A, lantrunculin D, actinomycin-D, Ro-31- 7453 (3-(6-nitro-1-methyl-3-indolyl)-4-(1-methyl-3-indolyl)pyrrole-2,5-dione), PNU-151807, brostallicin, C2-ceramide, cytarabine ocfosfate (2(1 H)- pyrimidinone, 4-amino-1-(5-0-(hydroxy(octadecyloxy)phosphinyl)- ⁇ -D- arabinofuranosyl)-, monosodium salt), paclitaxel (5 ⁇ ,20-epoxy-1 ,2 alpha,4,7 ⁇ ,10 ⁇ ,13 alpha-hexahydroxytax-11-en-9-one-4,10-diacetate-2- benzoate-13-(alpha
• the pharmacologically active fibrosis- inhibiting compound is a cyclin dependent protein kinase inhibitor (e.g., R- roscovitine, CYC-101 , CYC-103, CYC-400, MX-7065, alvocidib (4H-1- Benzopyran-4-one, 2-(2-chlorophenyl)-5,7-dihydroxy-8-(3-hydroxy-1 -methyI-4- piperidinyl)-, cis-(-)-), SU-9516, AG-12275, PD-0166285, CGP-79807, fascaplysin, GW-8510 (benzenesulfonamide, 4-((Z)-(6,7-dihydro-7-oxo-8H- pyrrolo(2,3-g)benzothiazol-8-ylidene)methyl)amino)-N-(3-hydroxy
• a cyclin dependent protein kinase inhibitor e.
• the pharmacologically active fibrosis- inhibiting compound is an EGF (epidermal growth factor) kinase inhibitor (e.g., erlotinib (4-quinazolinamine, N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-, monohydrochloride), erbstatin, BIBX-1382, gefitinib (4-quinazolinamine, N-(3- chloro-4-fluorophenyl)-7-methoxy-6-(3-(4-morpholinyl)propoxy)), or an analogue or derivative thereof).
• EGF epidermatiti factor
• the pharmacologically active fibrosis- inhibiting compound is an elastase inhibitor (e.g., ONO-6818, sivelestat sodium hydrate (glycine, N-(2-(((4-(2,2-dimethyl-1- oxopropoxy)phenyl)sulfonyl)amino)benzoyl)-), erdosteine (acetic acid, ((2-oxo- 2-((tetrahydro-2-oxo-3-thienyl)amino)ethyl)thio)-), MDL-100948A, MDL-104238 (N-(4-(4-morpholinylcarbonyl)benzoyl)-L-valyl-N'-(3,3,4,4,4-pentafluoro-1-(1- methylethyl)-2-oxobutyl)-L-2-azetamide), MDL-27324 (L-pro
• elastase inhibitor e.
• the pharmacologically active fibrosis- inhibiting compound is a factor Xa inhibitor (e.g., CY-222, fondaparinux sodium (alpha-D-glucopyranoside, methyl 0-2-deoxy-6-0-sulfo-2-(sulfoamino)-alpha-D- glucopyranosyl-(1-4)-0- ⁇ -D-glucopyranuronosyl-(1-4)-0-2-deoxy-3,6-di-0- sulfo-2-(sulfoamino)-alpha-D-glucopyranosyl-(1-4)-0-2-0-sulfo-alpha-L- idopyranuronosyl-(1 -4)-2-deoxy-2-(sulfoamino)-, 6-(hydrogen sulfate)), danaparoid sodium, or an analogue or derivative thereof).
• factor Xa inhibitor e.g., CY-222, fondapar
• the pharmacologically active fibrosis- inhibiting compound is a famesyltransferase inhibitor (e.g., dichlorobenzoprim (2,4-diamino-5-(4-(3,4-dichlorobenzylamino)-3-nitrophenyl)-6-ethylpyrimidine), B-581 , B-956 (N-(8(R)-amino-2(S)-benzyl-5(S)-isopropyl-9-sulfanyl-3(Z),6(E)- nonadienoyl)-L-methionine), OSI-754, perillyl alcohol (1-cyclohexene-1- methanol, 4-(1-methylethenyl)-, RPR-114334, lonafarnib (1- piperidinecarboxamide, 4-(2-(4-((11 R)-3,10-dibromo-8-ch
• the pharmacologically active fibrosis- inhibiting compound is a fibrinogen antagonist (e.g., 2(S)-((p- toluenesulfonyl)amino)-3-(((5,6,7,8,-tetrahydro-4-oxo-5-(2-(piperidin-4-yl)ethyl)- 4H-pyrazolo-(1 ,5-a)(1 ,4)diazepin-2-yl)carbonyl)-amino)propionic acid, streptokinase (kinase (enzyme-activating), strepto-), urokinase (kinase (enzyme-activating), uro-), plasminogen activator, pamiteplase, monteplase, heberkinase, anistreplase, alteplase, pro-urokinase, picotamide
• fibrinogen antagonist e.g., 2(
• the pharmacologically active fibrosis- inhibiting compound is a guanylate cyclase stimulant (e.g., isosorbide-5- mononitrate (D-glucitol, 1 ,4:3,6-dianhydro-, 5-nitrate), or an analogue or derivative thereof).
• a guanylate cyclase stimulant e.g., isosorbide-5- mononitrate (D-glucitol, 1 ,4:3,6-dianhydro-, 5-nitrate), or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a heat shock protein 90 antagonist (e.g., geldanamycin; NSC-33050 (17-allylaminogeldanamycin), rifabutin (rifamycin XIV, 1',4- didehydro-1-deoxy-1 ,4-dihydro-5'-(2-methylpropyl)-1-oxo-), 17AAG, or an analogue or derivative thereof).
• a heat shock protein 90 antagonist e.g., geldanamycin; NSC-33050 (17-allylaminogeldanamycin), rifabutin (rifamycin XIV, 1',4- didehydro-1-deoxy-1 ,4-dihydro-5'-(2-methylpropyl)-1-oxo-), 17AAG, or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is an HMGCoA reductase inhibitor (e.g., BCP-671 , BB- 476, fluvastatin (6-heptenoic acid, 7-(3-(4-fluorophenyl)-1-(1-methylethyl)-1 H- indol-2-yl)-3,5-dihydroxy-, monosodium salt, (R*,S*-(E))-( ⁇ )-), dalvastatin (2H- pyran-2-one, 6-(2-(2-(2-(4-fluoro-3-methylphenyl)-4,4,6,6-tetramethyl-1 - cyclohexen-1-yl)ethenyl)tetrahydro)-4-hydroxy-, (4alpha,6 ⁇ (E))-(+/-)-), glenvastatin (2H-pyran-2-one
• HMGCoA reductase inhibitor e.g.
• the pharmacologically active fibrosis- inhibiting compound is a hydroorotate dehydrogenase inhibitor (e.g., leflunomide (4-isoxazolecarboxamide, 5-methyl-N-(4-(trifluoromethyl)phenyl)-), laflunimus (2-propenamide, 2-cyano-3-cyclopropyl-3-hydroxy-N-(3-methyl- 4(trifluoromethyl)phenyl)-, (Z)-), or atovaquone (1 ,4-naphthalenedione, 2-(4-(4- chlorophenyl)cyclohexyl)-3-hydroxy-, trans-, or an analogue or derivative thereof).
• hydroorotate dehydrogenase inhibitor e.g., leflunomide (4-isoxazolecarboxamide, 5-methyl-N-(4-(trifluoromethyl)phenyl)-), laflunimus (2-propenamide, 2-cyano-3-cyclopropyl-3-
• the pharmacologically active fibrosis- inhibiting compound is an IKK2 inhibitor (e.g., MLN-120B, SPC-839, or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is an IL-1 , ICE or an IRAK antagonist (e.g., E-5090 (2- propenoic acid, 3-(5-ethyl-4-hydroxy-3-methoxy-1-naphthalenyl)-2-methyl-, (Z)- ), CH-164, CH-172, CH-490, AMG-719, iguratimod (N-(3-(formylamino)-4-oxo- 6-phenoxy-4H-chromen-7-yI) methanesulfonamide), AV94-88, pralnacasan (6H-pyridazino(1 ,2-a)(1 ,2)diazepine-1 -carboxamide, N-((2R,3S)-2- ethoxytetrahydro-5-oxo-3-furanyl)octahydro-9-((1 )
• an IRAK antagonist e.g., E-5090 (2
• the pharmacologically active fibrosis- inhibiting compound is an IL-4 agonist (e.g., glatiramir acetate (L-glutamic acid, polymer with L-alanine, L-lysine and L-tyrosine, acetate (salt)), or an analogue or derivative thereof).
• an IL-4 agonist e.g., glatiramir acetate (L-glutamic acid, polymer with L-alanine, L-lysine and L-tyrosine, acetate (salt)
• an analogue or derivative thereof e.g., glatiramir acetate (L-glutamic acid, polymer with L-alanine, L-lysine and L-tyrosine, acetate (salt)
• the pharmacologically active fibrosis- inhibiting compound is an immunomodulatory agent (e.g., biolimus, ABT-578, methylsulfamic acid 3-(2-methoxyphenoxy)-2-
• an immunomodulatory agent e.g., biolimus, ABT-578, methylsulfamic acid 3-(2-methoxyphenoxy)-2-
• analogues of rapamycin include tacrolimus and derivatives thereof (e.g., EP0184162B1 and U.S. Patent No. 6,258,823) everolimus and derivatives thereof (e.g., U.S. Patent No. 5,665,772). Further representative examples of sirolimus analogues and derivatives can be found in PCT Publication Nos.
• Representative U.S. patents include U.S. Patent Nos. 6,342,507; 5,985,890; 5,604,234;
• sirolimus analogues and derivatives include tacrolimus and derivatives thereof (e.g., EP0184162B1 and U.S. Patent No. 6,258,823) everolimus and derivatives thereof (e.g., US Patent No. 5,665,772).
• Further representative examples of sirolimus analogues and derivatives include ABT- 578 and others may be found in PCT Publication Nos.
• WO 97/10502 WO 96/41807, WO 96/35423, WO 96/03430, WO 9600282, WO 95/16691 , WO 9515328, WO 95/07468, WO 95/04738, WO 95/04060, WO 94/25022, WO 94/21644, WO 94/18207, WO 94/10843, WO 94/09010, WO 94/04540, WO 94/02485, WO 94/02137, WO 94/02136, WO 93/25533, WO 93/18043, WO 93/13663, WO 93/11130, WO 93/10122, WO 93/04680, WO 92/14737, and WO 92/05179.
• U.S. patents include U.S. Patent Nos. 6,342,507; 5,985,890; 5,604,234; 5,597,715; 5,583,139; 5,563,172; 5,561 ,228; 5,561 ,137 5,541 ,193; 5,541 ,189; 5,534,632; 5,527,907; 5,484,799; 5,457,194; 5,457,182 5,362,735; 5,324,644; 5,318,895; 5,310,903; 5,310,901 ; 5,258,389; 5,252,732 5,247,076; 5,225,403; 5,221 ,625; 5,210,030; 5,208,241 , 5,200,411 ; 5,198,421 5,147,877; 5,140,018; 5,116,756; 5,109,112; 5,093,338; and 5,091 ,389.
• the fibrosis-inhibiting agent may be, e.g., rapamycin (sirolimus), everolimus, biolimus, tresperimus, auranofin, 27-0- demethylrapamycin, tacrolimus, gusperimus, pimecrolimus, or ABT-578.
• the pharmacologically active fibrosis- inhibiting compound is an inosine monophosphate dehydrogenase (IMPDH) inhibitor (e.g., mycophenolic acid, mycophenolate mofetil (4-hexenoic acid, 6- (1 ,3-dihydro-4-hydroxy-6-methoxy-7-methyl-3-oxo-5-isobenzofuranyl)-4-methyl- , 2-(4-morpholinyl)ethyl ester, (E)-), ribavirin (1 H-1 ,2,4-triazole-3-carboxamide, 1 - ⁇ -D-ribof uranosyl-), tiazofurin (4-thiazolecarboxamide, 2- ⁇ -D-ribofuranosyl-), viramidine, aminothiadiazole, thiophenfurin, tiazofurin) or an analogue or derivative thereof.
• IMPDH inosine monophosphate dehydrogenase
• the pharmacologically active fibrosis- inhibiting compound is a leukotreine inhibitor (e.g., ONO- 4057(benzenepropanoic acid, 2-(4-carboxybutoxy)-6-((6-(4-methoxyphenyl)-5- hexenyl)oxy)-, (E)-), ONO-LB-448, pirodomast 1 ,8-naphthyridin-2(1 H)-one, 4- hydroxy-1-phenyl-3-(1-pyrrolidinyl)-, Sch-40120 (benzo(b)(1 ,8)naphthyridin- 5(7H)-one, 10-(3-chlorophenyl)-6,8,9,10-tetrahydro-), L-656224 (4- benzofuranol, 7-chloro-2-((4-methoxyphenyl)methyl)-3-methyl-5-prop
• a leukotreine inhibitor e.
• the pharmacologically active fibrosis- inhibiting compound is a MCP-1 antagonist (e.g., nitronaproxen (2- napthaleneacetic acid, 6-methoxy-alpha-methyl 4-(nitrooxy)butyl ester (alpha S)-), bindarit (2-(1-benzylindazol-3-ylmethoxy)-2-methylpropanoic acid), 1- alpha-25 dihydroxy vitamin D 3 ⁇ or an analogue or derivative thereof).
• MCP-1 antagonist e.g., nitronaproxen (2- napthaleneacetic acid, 6-methoxy-alpha-methyl 4-(nitrooxy)butyl ester (alpha S)-), bindarit (2-(1-benzylindazol-3-ylmethoxy)-2-methylpropanoic acid), 1- alpha-25 dihydroxy vitamin D 3 ⁇ or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is a matrix metalloproteinase (MMP) inhibitor (e.g., D- 9120, doxycycline (2-naphthacenecarboxamide, 4-(dimethylamino)- 1 ,4,4a,5,5a,6,11 ,12a-octahydro-3,5,10,12,12a-pentahydroxy-6-methyl-1 ,11- dioxo- (4S-(4 alpha, 4a alpha, 5 Ipha, 5a alpha, 6 alpha, 12a alpha))-), BB- 2827, BB-1101 (2S-allyl-N1-hydroxy-3R-isobutyl-N4-(1S-methylcarbamoyl-2- phenylethyl)-succinamide), BB-2983, solimastat (N'-(2,2-dimethyl-1(S)-(N-(MMP) inhibitor) inhibitor (e.g
• the pharmacologically active fibrosis- inhibiting compound is a NF kappa B (NFKB) inhibitor (e.g., AVE-0545, Oxi-104 (benzamide, 4-amino-3-chloro-N-(2-(diethylamino)ethyl)-), dexlipotam, R- flurbiprofen ((1 ,1'-biphenyl)-4-acetic acid, 2-fluoro-alpha-methyl), SP100030 (2- chloro-N-(3,5-di(trifluoromethyl)phenyl)-4-(trifluoromethyl)pyrimidine-5- carboxamide), AVE-0545, Viatris, AVE-0547, Bay 11-7082, Bay 11-7085, 15 deoxy-prostaylandin J2, bortezomib (boronic acid, ((1 R)-3-methyl-1-(((2S)-1
• NFKB NF kappa B
• the pharmacologically active fibrosis- inhibiting compound is a NO antagonist (e.g., NCX-4016 (benzoic acid, 2- (acetyloxy)-, 3-((nitrooxy)methyl)phenyI ester, NCX-2216, L-arginine or an analogue or derivative thereof).
• NO antagonist e.g., NCX-4016 (benzoic acid, 2- (acetyloxy)-, 3-((nitrooxy)methyl)phenyI ester, NCX-2216, L-arginine or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a p38 MAP kinase inhibitor (e.g., GW-2286, CGP-52411 , BIRB-798, SB220025, RO-320-1195, RWJ-67657, RWJ-68354, SCIO-469, SCIO-323, AMG-548, CMC-146, SD-31145, CC-8866, Ro-320-1195, PD-98059 (4H-1 -benzopyran-4-one, 2-(2-amino-3-methoxyphenyl)-), CGH-2466, doramapimod, SB-203580 (pyridine, 4-(5-(4-fluorophenyl)-2-(4-
• WO 00/63204A2 WO 01/21591A1 ; WO 01/35959A1 ; WO 01/74811A2; WO 02/18379A2; WO 2064594A2; WO 2083622A2; WO 2094842A2; WO 2096426A1 ; WO 2101015A2; WO 2103000A2; WO 3008413A1 ; WO 3016248A2; WO 3020715A1 ; WO 3024899A2; WO 3031431A1 ; WO3040103A1 ; WO 3053940A1 ; WO 3053941 A2; WO 3063799A2; WO 3079986A2; WO 3080024A2; WO 3082287A1 ; WO 97/44467A1 ; WO 99/01449A1 ; and WO 99/58523A1.
• the pharmacologically active fibrosis- inhibiting compound is a phosphodiesterase inhibitor (e.g., CDP-840 (pyridine, 4-((2R)-2-(3-(cyclopentyloxy)-4-methoxyphenyl)-2-phenylethyl)-), CH-3697, CT- 2820, D-22888 (imidazo(1 ,5-a)pyrido(3,2-e)pyrazin-6(5H)-one, 9-ethyl-2- methoxy-7-methyl-5-propyl-), D-4418 (8-methoxyquinoline-5-(N-(2,5- dichIoropyridin-3-yl))carboxamide), 1-(3-cyclopentyloxy-4-methoxyphenyl)-2- (2,6-dichloro-4-pyridyl) ethanone oxime, D-4396, ONO-6126, CDC-998,
• CDP-840 pyridine, 4-((2
• phosphodiesterase inhibitors include denbufylline (1 H-purine-2,6-dione, 1 ,3-dibutyl-3,7-dihydro-7-(2-oxopropyl)-), propentofylline (1 H-purine-2,6-dione, 3,7-dihydro-3-methyl-1-(5-oxohexyl)-7- propyl-) and pelrinone (5-pyrimidinecarbonitrile, 1 ,4-dihydro-2-methyl-4-oxo-6- ((3-pyridinylmethyl)amino)-).
• phosphodiesterase III inhibitors include enoximone (2H-imidazol-2-one, 1 ,3-dihydro-4-methyl-5-(4-(methylthio)benzoyl)- ), and saterinone (3-pyridinecarbonitrile, 1 ,2-dihydro-5-(4-(2-hydroxy-3-(4-(2- methoxyphenyl)-1-piperazinyl)propoxy)phenyl)-6-methyl-2-oxo-).
• phosphodiesterase IV inhibitors include AWD- 12-281 , 3-auinolinecarboxylic acid, 1 -ethyl-6-fIuoro-1 ,4-dihydro-7-(4-methyl-1 - piperazinyl)-4-oxo-), tadalafil (pyrazino(1',2':1 ,6)pyrido(3,4-b)indole1 ,4-dione, 6- (1 ,3-benzodioxol-5-yI)-2,3,6,7,12,12a-hexahydro-2-methyl-, (6R-trans)), and filaminast (ethanone, 1-(3-(cyclopentyloxy)-4-methoxyphenyl)-, O- (aminocarbonyl)oxime, (1 E)-)
• Another example of a phosphodiesterase V inhibitor is vardenafil
• the pharmacologically active fibrosis- inhibiting compound is a TGF beta Inhibitor (e.g., mannose-6-phosphate, LF- 984, tamoxifen (ethanamine, 2-(4-(1 ,2-diphenyl-1-butenyl)phenoxy)-N,N- dimethyl-, (Z)-), tranilast, or an analogue or derivative thereof).
• a TGF beta Inhibitor e.g., mannose-6-phosphate, LF- 984, tamoxifen (ethanamine, 2-(4-(1 ,2-diphenyl-1-butenyl)phenoxy)-N,N- dimethyl-, (Z)-), tranilast, or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a thromboxane A2 antagonist (e.g., CGS-22652 (3- pyridineheptanoic acid, ⁇ -(4-(((4-chlorophenyl)sulfonyl)amino)butyl)-, (.+-.)-), ozagrel (2-propenoic acid, 3-(4-(1 H-imidazol-1-ylmethyl)phenyl)-, (E)-), argatroban (2-piperidinecarboxylic acid, 1-(5-((aminoiminomethyl)amino)-1-oxo- 2-(((1 ,2,3,4-tetrahydro-3-methyl-8-quinolinyl)sulfonyl)amino)pentyl)-4-methyl-), ramatroban (9H-carbazole-9-propanoic acid,
• the pharmacologically active fibrosis- inhibiting compound is a tyrosine kinase inhibitor (e.g., SKI-606, ER-068224, SD-208, N-(6-benzothiazolyl)-4-(2-(1-piperazinyl)pyrid-5-yl)-2-pyrimidineamine, celastrol (24,25,26-trinoroleana-1(10),3,5,7-tetraen-29-oic acid, 3-hydroxy-9,13- dimethyl-2-oxo-, (9 beta.,13alpha,14 ⁇ ,20 alpha)-), CP-127374 (geldanamycin, 17-demethoxy-17-(2-propenylamino)-), CP-564959, PD-171026, CGP-52411 (1 H-lsoindole-1 ,3(2H)-dione, 4,5-bis(phenylamino)
• tyrosine kinase inhibitor e
• the pharmacologically active fibrosis- inhibiting compound is a vitronectin inhibitor (e.g., O-(9,10-dimethoxy- 1 ,2, 3,4,5, 6-hexahydro-4-((1 , 4,5, 6-tetrahydro-2-pyrimidinyl)hydrazono)-8- benz(e)azulenyl)-N-((phenylmethoxy)carbonyl)-DL-homoserine 2,3- dihydroxypropyl ester, (2S)-benzoylcarbonylamino-3-(2-((4S)-(3-(4,5-dihydro- 1 H-imidazol-2-ylamino)-propyl)-2,5-dioxo-imidazolidin-1 -yl)-acetylamino)- propionate, Sch-221153, S-836, SC-68448 ( ⁇ -((2-2-(((2-2-(((2--(((2-2-(
• the pharmacologically active fibrosis- inhibiting compound is a fibroblast growth factor inhibitor (e.g., CT-052923 (((2H-benzo(d)1 ,3-dioxalan-5-methyl)amino)(4-(6,7-dimethoxyquinazolin-4- yl)piperazinyl)methane-1 -thione), or an analogue or derivative thereof).
• a fibroblast growth factor inhibitor e.g., CT-052923 (((2H-benzo(d)1 ,3-dioxalan-5-methyl)amino)(4-(6,7-dimethoxyquinazolin-4- yl)piperazinyl)methane-1 -thione
• the pharmacologically active fibrosis- inhibiting compound is a protein kinase inhibitor (e.g., KP-0201448, NPC15437 (hexanamide, 2,6-diamino-N-((1-(1-oxotridecyl)-2-piperidinyl)methyl)-), fasudil (1 H-1 ,4-diazepine, hexahydro-1-(5-isoquinolinylsulfonyl)-), midostaurin (benzamide, N-(2,3,10,11 ,12,13-hexahydro-10-methoxy-9-methyl-1-oxo-9,13- epoxy-1 H,9H-diindolo(1 ,2,3-gh:3 , ,2',1 , -lm)pyrrolo(3,4-j)(1 ,7)benzodiazonin-11- y
• a protein kinase inhibitor e
• the pharmacologically active fibrosis- inhibiting compound is a PDGF receptor kinase inhibitor (e.g., RPR-127963E, or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is an endothelial growth factor receptor kinase inhibitor (e.g., CEP-7055, SU-0879 ((E)-3-(3,5-di-tert-butyl-4-hydroxyphenyl)-2- (aminothiocarbonyl)acrylonitrile), BIBF-1000, AG-013736 (CP-868596), AMG- 706, AVE-0005, NM-3 (3-(2-methylcarboxymethyl)-6-methoxy-8-hydroxy- isocoumarin), Bay-43-9006, SU-011248, or an analogue or derivative thereof).
• endothelial growth factor receptor kinase inhibitor e.g., CEP-7055, SU-0879 ((E)-3-(3,5-di-tert-butyl-4-hydroxyphenyl)-2- (aminothiocarbonyl)acrylonitrile), BIBF-1000, AG-013736 (CP
• the pharmacologically active fibrosis- inhibiting compound is a retinoic acid receptor antagonist (e.g., etarotene (Ro- 15-1570) (naphthalene, 6-(2-(4-(ethylsulfonyl)phenyl)-1-methylethenyl)-1 ,2,3,4- tetrahydro-1 ,1 ,4,4-tetramethyl-, (E)-), (2E,4E)-3-methyl-5-(2-((E)-2-(2,6,6- trimethyl-1 -cyclohexen-1 -yl)ethenyl)-1 -cyclohexen-1 -yl)-2,4-pentadienoic acid, tocoretinate (retinoic acid, 3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12- trimethyltridecyl)-2H-1-benzo
• retinoic acid receptor antagonist e.g., e
• the pharmacologically active fibrosis- inhibiting compound is a platelet derived growth factor receptor kinase inhibitor (e.g., leflunomide (4-isoxazolecarboxamide, 5-methyl-N-(4- (trifluoromethyl)phenyl)-, or an analogue or derivative thereof).
• a platelet derived growth factor receptor kinase inhibitor e.g., leflunomide (4-isoxazolecarboxamide, 5-methyl-N-(4- (trifluoromethyl)phenyl)-, or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a fibrinogin antagonist (e.g., picotamide (1 ,3- benzenedicarboxamide, 4-methoxy-N,N'-bis(3-pyridinylmethyl)-, or an analogue or derivative thereof).
• a fibrinogin antagonist e.g., picotamide (1 ,3- benzenedicarboxamide, 4-methoxy-N,N'-bis(3-pyridinylmethyl)-, or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is an antimycotic agent (e.g., miconazole, sulconizole, parthenolide, rosconitine, nystatin, isoconazole, fluconazole, ketoconasole, imidazole, itraconazole, terpinafine, elonazole, bifonazole, clotrimazole, conazole, terconazole (piperazine, 1-(4-((2-(2,4-dichlorophenyl)-2-(1 H-1 ,2,4- triazol-1 -ylmethyl)-1 ,3-dioxolan-4-yl)methoxy)phenyl)-4-(1 -methylethyl)-, cis-), isoconazole (1-(2-(2-6-dichlorobenzyloxy)-2-(2-,4-dichIoroph
• an antimycotic agent e.g
• the pharmacologically active fibrosis- inhibiting compound is a bisphosphonate (e.g., clodronate, alendronate, pamidronate, zoledronate, or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is a phospholipase A1 inhibitor (e.g., ioteprednol etabonate (androsta-1 ,4-diene-17-carboxylic acid, 17-((ethoxycarbonyl)oxy)-11 -hydroxy-3- oxo-, chloromethyl ester, (11 ⁇ ,17 alpha)-, or an analogue or derivative thereof).
• a phospholipase A1 inhibitor e.g., ioteprednol etabonate (androsta-1 ,4-diene-17-carboxylic acid, 17-((ethoxycarbonyl)oxy)-11 -hydroxy-3- oxo-, chloromethyl ester, (11 ⁇ ,17 alpha)-, or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a histamine H1 , H2, or H3 receptor antagonist (e.g., ranitidine (1 ,1-ethenediamine, N-(2-(((5-((dimethylamino)methyl)-2- furanyl)methyl)thio)ethyl)-N'-methyl-2-nitro-), niperotidine (N-(2-((5- ((dimethylamino)methyl)furfuryl)thio)ethyl)-2-nitro-N'-piperonyl-1 ,1- ethenediamine), famotidine (propanimidamide, 3-(((2- ((aminoiminomethyl)amino)-4-thiazolyl)methyl)thio)-N-(aminosulfonyl)-), roxitadine acetate HCI (acetamide, 2-(acetamide, 2-(acetamide, 2-(acetamide, 2-(acety
• the pharmacologically active fibrosis- inhibiting compound is a macrolide antibiotic (e.g., dirithromycin (erythromycin, 9-deoxo-11-deoxy-9,11-(imino(2-(2-methoxyethoxy)ethylidene)oxy)-, (9S(R))-), flurithromycin ethylsuccinate (erythromycin, 8-fluoro-mono(ethyl butanedioate) (ester)-), erythromycin stinoprate (erythromycin, 2'-propanoate, compound with N-acetyl-L-cysteine (1 :1)), clarithromycin (erythromycin, 6-O-methyl-), azithromycin (9-deoxo-9a-aza-9a-methyl-9a-homoerythromycin-A), telithromycin (3-de((2,6-dideoxy-3-C-methyl-3-0-methyl-methyl-
• a macrolide antibiotic
• the pharmacologically active fibrosis- inhibiting compound is a GPIIb Ilia receptor antagonist (e.g., tirofiban hydrochloride (L-tyrosine, N-(butylsulfonyl)-0-(4-(4-piperidinyl)butyl)-, monohydrochloride-), eptifibatide (L-cysteinamide, N6-(aminoiminomethyl)-N2- (3-mercapto-1-oxopropyl)-L-lysylglycyl-L-alpha-aspartyl-L-tryptophyl-L-prolyl-, cyclic(1->6)-disulfide), xemilofiban hydrochloride, or an analogue or derivative thereof).
• a GPIIb Ilia receptor antagonist e.g., tirofiban hydrochloride (L-tyrosine, N-(butylsulfonyl)-0-
• the pharmacologically active fibrosis- inhibiting compound is an endothelin receptor antagonist (e.g., bosentan (benzenesulfonamide, 4-(1 ,1-dimethylethyI)-N-(6-(2-hydroxyethoxy)-5-(2- methoxyphenoxy)(2,2 , -bipyrimidin)-4-yl)-, or an analogue or derivative thereof).
• an endothelin receptor antagonist e.g., bosentan (benzenesulfonamide, 4-(1 ,1-dimethylethyI)-N-(6-(2-hydroxyethoxy)-5-(2- methoxyphenoxy)(2,2 , -bipyrimidin)-4-yl
• bosentan benzenesulfonamide, 4-(1 ,1-dimethylethyI)-N-(6-(2-hydroxyethoxy)-5-(2- methoxyphenoxy)(2,2 , -
• the pharmacologically active fibrosis- inhibiting compound is a peroxisome proliferator-activated receptor agonist (e.g., gemfibrozil (pentanoic acid, 5-(2,5-dimethylphenoxy)-2,2-dimethyl-), fenofibrate (propanoic acid, 2-(4-(4-chlorobenzoyl)phenoxy)-2-methyl-, 1- methylethyl ester), ciprofibrate (propanoic acid, 2-(4-(2,2- dichlorocyclopropyl)phenoxy)-2-methyl-), rosiglitazone maleate (2,4- thiazolidinedione, 5-((4-(2-(methyl-2-pyridinylamino)ethoxy)phenyl)methyl)-, (Z)- 2-butenedioate (1 :1 )), pioglitazone hydrochlor
• the pharmacologically active compound is a peroxisome proliferator-activated receptor alpha agonist, such as GW-590735, GSK-677954, GSK501516, pioglitazone hydrochloride (2,4-thiazolidinedione, 5- ((4-(2-(5-ethyl-2-pyridinyl)ethoxy)phenyl)methyl)-, monohydrochloride (+/-)-, or an analogue or derivative thereof).
• Estrogen Receptor Agents In another embodiment, the pharmacologically active fibrosis- inhibiting compound is an estrogen receptor agent (e.g., estradiol, 17- ⁇ - estradiol, or an analogue or derivative thereof).
• Somatostatin Analogues In another embodiment, the pharmacologically active fibrosis- inhibiting compound is a somatostatin analogue (e.g., angiopeptin, or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is a neurokinin 1 antagonist (e.g., GW-597599, lanepitant ((1 ,4'-bipiperidine)-1 '-acetamide, N-(2-(acetyl((2-methoxyphenyl)methyl)amino)- 1-(1H- ⁇ ndol-3-ylmethyl)ethyl)- (R)-), nolpitantium chloride (1- azoniabicyclo(2.2.2)octane, 1-(2-(3-(3,4-dichlorophenyl)-1-((3-(1- methylethoxy)phenyl)acetyl)-3-piperidinyl)ethyl)-4-phenyl-, chloride, (S)-), or saredutant (benzamide, N-(4-(4-(acetylamino)-4-phenyl-1
• a neurokinin 1 antagonist e.
• the pharmacologically active fibrosis- inhibiting compound is a neurokinin 3 antagonist (e.g., talnetant (4- quinolinecarboxamide, 3-hydroxy-2-phenyl-N-((1S)-1-phenylpropyl)-, or an analogue or derivative thereof).
• a neurokinin 3 antagonist e.g., talnetant (4- quinolinecarboxamide, 3-hydroxy-2-phenyl-N-((1S)-1-phenylpropyl)-, or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a neurokinin antagonist (e.g., GSK-679769, GSK- 823296, SR-489686 (benzamide, N-(4-(4-(acetylamino)-4-phenyl-1-piperidinyl)- 2-(3,4-dichlorophenyl)butyl)-N-methyl-, (S)-), SB-223412; SB-235375 (4- quinolinecarboxamide, 3-hydroxy-2-phenyl-N-((1S)-1-phenylpropyl)-), UK- 226471 , or an analogue or derivative thereof).
• a neurokinin antagonist e.g., GSK-679769, GSK- 823296, SR-489686 (benzamide, N-(4-(4-(acetylamino)-4-phenyl-1-piperidinyl)- 2-(3,4-dichlorophenyl)butyl
• VLA-4 Antagonist in another embodiment, is a VLA-4 antagonist (e.g., GSK683699, or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is a osteoclast inhibitor (e.g., ibandronic acid (phosphonic acid, (1-hydroxy-3-(methylpentylamino)propylidene) bis-), alendronate sodium, or an analogue or derivative thereof).
• a osteoclast inhibitor e.g., ibandronic acid (phosphonic acid, (1-hydroxy-3-(methylpentylamino)propylidene) bis-
• alendronate sodium or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a DNA topoisomerase ATP hydrolyzing inhibitor (e.g., enoxacin (1 ,8-naphthyridine-3-carboxylic acid, 1 -ethyl-6-fluoro-1 ,4-dihydro-4- oxo-7-(1 -piperazinyl)-), levofloxacin (7H-Pyrido(1 ,2,3-de)-1 ,4-benzoxazine-6- carboxylic acid, 9-fluoro-2,3-dihydro-3-methyl-10-(4-methyl-1-piperazinyl)-7- oxo-, (S)-), ofloxacin (7H-pyrido(1 ,2,3-de)-1 ,4-benzoxazine-6-carboxyIic acid, 9- fluoro
• a DNA topoisomerase ATP hydrolyzing inhibitor e.g.,
• the pharmacologically active fibrosis- inhibiting compound is an angiotensin I converting enzyme inhibitor (e.g., ramipril (cyclopenta(b)pyrrole-2-carboxylic acid, 1-(2-((1-(ethoxycarbonyl)-3- phenylpropyl)amino)-1-oxopropyl)octahydro-, (2S-(1(R*(R*)),2 alpha, 3a ⁇ , 6a ⁇ ))-), trandolapril (1 H-indole-2-carboxylic acid, 1-(2-((1-carboxy-3- phenylpropyl)amino)-1 -oxopropyl)octahydro-, (2S-(1 (R*(R*)),2 alpha,3a alpha,7a ⁇ ))-), fasidotril
• an angiotensin I converting enzyme inhibitor e.g., ram
• the pharmacologically active fibrosis- inhibiting compound is an angiotensin II antagonist (e.g., HR-720 (1 H- imidazole-5-carboxylic acid, 2-butyl-4-(methylthio)-1-((2'-)
• angiotensin II antagonist e.g., HR-720 (1 H- imidazole-5-carboxylic acid, 2-butyl-4-(methylthio)-1-((2'-
• the pharmacologically active fibrosis- inhibiting compound is an enkephalinase inhibitor (e.g., Aventis 100240 (pyrido(2,1-a)(2)benzazepine-4-carboxylic acid, 7-((2-(acetylthio)-1 -oxo-3- phenyIpropyl)amino)-1 ,2,3,4,6,7,8,12b-octahydro-6-oxo-, (4S-(4 alpha, 7 alpha(R*),12b ⁇ ))-), AVE-7688, or an analogue or derivative thereof).
• Aventis 100240 pyrido(2,1-a)(2)benzazepine-4-carboxylic acid, 7-((2-(acetylthio)-1 -oxo-3- phenyIpropyl)amino)-1 ,2,3,4,6,7,8,12b-octahydro-6-oxo-, (4S-(4 al
• the pharmacologically active fibrosis- inhibiting compound is peroxisome proliferator-activated receptor gamma agonist insulin sensitizer (e.g., rosiglitazone maleate (2,4-thiazolidinedione, 5- ((4-(2-(methyl-2-pyridinylamino)ethoxy)phenyl)methyl)-, (Z)-2-butenedioate (1 :1 ), farglitazar (GI-262570, GW-2570, GW-3995, GW-5393, GW-9765), LY- 929, LY-519818, LY-674, or LSN-862), or an analogue or derivative thereof).
• peroxisome proliferator-activated receptor gamma agonist insulin sensitizer e.g., rosiglitazone maleate (2,4-thiazolidinedione, 5- ((4-(2-(methyl-2-pyridinylamino)ethoxy)phenyl)
• the pharmacologically active fibrosis- inhibiting compound is a protein kinase C inhibitor, such as ruboxistaurin mesylate (9H,18H-5,21 :12,17-dimethenodibenzo(e,k)pyrrolo(3,4- h)(1 , 4,13)oxadiazacyclohexadecine-18,20(19H)-dione, 9- ((dimethylamino)methyl)-6,7,10,11-tetrahydro-, (S)-), safingol (1 ,3- octadecanediol, 2-amino-, (S-(R*,R*)) ⁇ ), or enzastaurin hydrochloride (1 H- pyrole-2,5-dione, 3-(1 -methyl-1 H-indol-3-yl)-4-(1 -(1 -(2-a protein kinase C inhibitor, such as ruboxistauri
• the pharmacologically active fibrosis- inhibiting compound is a ROCK (rho-associated kinase) inhibitor, such as Y- 27632, HA-1077, H-1152 and 4-1-(aminoalkyl)-N-(4-pyridyl) cyclohexanecarboxamide or an analogue or derivative thereof.
• a ROCK (rho-associated kinase) inhibitor such as Y- 27632, HA-1077, H-1152 and 4-1-(aminoalkyl)-N-(4-pyridyl) cyclohexanecarboxamide or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a CXCR3 inhibitor such as T-487, T0906487 or analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is an Itk inhibitor such as BMS-509744 or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a cytosolic phospholipase A 2 -alpha inhibitor such as efipladib (PLA-902) or analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a PPAR Agonist (e.g., Metabolex ((-)-benzeneacetic acid, 4-chloro-alpha-(3-(trifluoromethyl)-phenoxy)-, 2-(acetylamino)ethyl ester), balaglitazone (5-(4-(3-methyl-4-oxo-3,4-dihydro-quinazolin-2-yl-methoxy)- benzyl)-thiazolidine-2,4-dione), ciglitazone (2,4-thiazolidinedione, 5-((4-((1- methylcyclohexyl)methoxy)phenyl)methyl)-), DRF-10945, farglitazar, GSK- 677954, GW-409544, GW-501516, GW-590735, GW-590735, K
• the pharmacologically active fibrosis- inhibiting compound is an immunosuppressant (e.g., batebulast (cyclohexanecarboxylic acid, 4-(((aminoiminomethyl)amino)methyl)-, 4-(1 ,1- dimethylethyl)phenyl ester, trans-), cyclomunine, exalamide (benzamide, 2- (hexyloxy)-), LYN-001 , CCI-779 (rapamycin 42-(3-hydroxy-2-(hydroxymethyl)-2- methylpropanoate)), 1726; 1726-D; AVE-1726, or an analogue or derivative thereof).
• an immunosuppressant e.g., batebulast (cyclohexanecarboxylic acid, 4-(((aminoiminomethyl)amino)methyl)-, 4-(1 ,1- dimethylethyl)phenyl ester, trans-), cyclomunine, exalamide (benzamide
• the pharmacologically active fibrosis- inhibiting compound is an Erb inhibitor (e.g., canertinib dihydrochloride (N-(4-(3- (chloro-4-fluoro-phenylamino)-7-(3-morpholin-4-yl-propoxy)-quinazolin-6-yl)- acrylamide dihydrochloride), CP-724714, or an analogue or derivative thereof).
• an Erb inhibitor e.g., canertinib dihydrochloride (N-(4-(3- (chloro-4-fluoro-phenylamino)-7-(3-morpholin-4-yl-propoxy)-quinazolin-6-yl)- acrylamide dihydrochloride), CP-724714, or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is an apoptosis agonist (e.g., CEFLATONIN (CGX-635) (from Chemgenex Therapeutics, Inc., Menlo Park, CA), CHML, LBH-589, metoclopramide (benzamide, 4-amino-5-chloro-N-(2-(diethylamino)ethyl)-2- methoxy-), patupilone (4,17-dioxabicyclo(14.1.0)heptadecane-5,9-dione, 7,11- dihydroxy-8,8,10,12,16-pentamethyl-3-(1-methyl-2-(2-methyl-4- thiazolyl)ethenyl, (1 R,3S,7S,10R,11S,12S,16R)), AN-9; pivanex (butanoic acid, (2,2-dimethyl-1-oxopropoxy)
• apoptosis agonist e.g
• the pharmacologically active fibrosis- inhibiting compound is an lipocortin agonist (e.g., CGP-13774 (9Alpha-chloro- 6Alpha-fluoro-11 ⁇ ,17alpha-dihydroxy-16Alpha-methyl-3-oxo-1 ,4-androstadiene- 17 ⁇ -carboxylic acid-methylester-17-propionate), or analogue or derivative thereof).
• an lipocortin agonist e.g., CGP-13774 (9Alpha-chloro- 6Alpha-fluoro-11 ⁇ ,17alpha-dihydroxy-16Alpha-methyl-3-oxo-1 ,4-androstadiene- 17 ⁇ -carboxylic acid-methylester-17-propionate
• VCAM-1 antagonist in another embodiment, is a VCAM-1 antagonist (e.g., DW-908e, or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is a collagen antagonist (e.g., E-5050 (Benzenepropanamide, 4-(2,6-dimethylheptyl)-N-(2-hydroxyethyl)- ⁇ -methyl-), lufironil (2,4-Pyridinedicarboxamide, N,N'-bis(2-methoxyethyl)-), or an analogue or derivative thereof).
• a collagen antagonist e.g., E-5050 (Benzenepropanamide, 4-(2,6-dimethylheptyl)-N-(2-hydroxyethyl)- ⁇ -methyl-), lufironil (2,4-Pyridinedicarboxamide, N,N'-bis(2-methoxyethyl)-
• E-5050 Benzenepropanamide, 4-(2,6-dimethylheptyl)-N-(2-hydroxyethyl)- ⁇ -methyl-
• lufironil (2,
• the pharmacologically active fibrosis- inhibiting compound is an alpha 2 integrin antagonist (e.g., E-7820, or an analogue or derivative thereof).
• the pharmacologically active fibrosis- inhibiting compound is a TNF alpha inhibitor (e.g., ethyl pyruvate, Genz-29155, lentinan (Ajinomoto Co., Inc.
• linomide 3-quinolinecarboxamide, 1 ,2- dihydro-4-hydroxy-N,1-dimethyl-2-oxo-N-phenyl-), UR-1505, or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a nitric oxide inhibitor (e.g., guanidioethyldisulfide, or an analogue or derivative thereof).
• a nitric oxide inhibitor e.g., guanidioethyldisulfide, or an analogue or derivative thereof.
• the pharmacologically active fibrosis- inhibiting compound is a cathepsin inhibitor (e.g., SB-462795 or an analogue or derivative thereof).
• the present invention also provides for the combination of a polymeric composition and an agent which reduces the likelihood of infection upon implantation of the composition or a medical implant.
• Infection is a common complication of the implantation of foreign bodies such as, for example, medical devices and implants.
• Foreign materials provide an ideal site for micro- organisms to attach and colonize. It is also hypothesized that there is an impairment of host defenses to infection in the microenvironment surrounding a foreign material. These factors make medical implants particularly susceptible to infection and make eradication of such an infection difficult, if not impossible, in most cases. In many cases, an infected implant or device must be surgically removed from the body in order to irradicate the infection.
• the present invention provides agents (e.g., chemotherapeutic agents) that can be released from a composition, and which have potent antimicrobial activity at extremely low doses.
• agents e.g., chemotherapeutic agents
• a wide variety of anti-infective agents can be utilized in combination with the present compositions. Suitable anti-infective agents may be readily determined based upon the assays provided in Example 34).
• agents that can be used as anti-infective agents such as: (A) anthracyclines (e.g., doxorubicin and mitoxantrone), (B) fluoropyrimidines (e.g., 5-FU), (C) folic acid antagonists (e.g., methotrexate), (D) podophylotoxins (e.g., etoposide), (E) camptothecins, (F) hydroxyureas, and (G) platinum complexes (e.g., cisplatin).
• anthracyclines e.g., doxorubicin and mitoxantrone
• fluoropyrimidines e.g., 5-FU
• C folic acid antagonists (e.g., methotrexate)
• D podophylotoxins
• E camptothecins
• F hydroxyureas
• platinum complexes e.g., cisplatin
• the therapeutic anti-infective agent is an anthracycline.
• Anthracyclines have the following general structure, where the R groups may be a variety of organic groups:
• R ⁇ is CH 3 or CH 2 OH
• R 2 is daunosamine or H
• R 3 and R 4 are independently one of OH, N0 2 , NH 2 , F, CI, Br, I, CN, H or groups derived from these
• R 5 is hydrogen, ydroxyl, or methoxy
• R 6- s are all hydrogen.
• R 5 and R 6 are hydrogen and R and R 8 are alkyl or halogen, or vice versa.
• R-i may be a conjugated peptide.
• R 5 may be an ether linked alkyl group.
• R 5 may be OH or an ether linked alkyl group.
• Ri may also be linked to the anthracycline ring by a group other than C(O), such as an alkyl or branched alkyl group having the C(O) linking moiety at its end, such as -CH 2 CH(CH 2 -X)C(0)-R ⁇ , wherein X is H or an alkyl group (see, e.g., U.S. Patent 4,215,062).
• R 3 may have the following structure:
• R- t o may be H or form a secondary amine with a group such as an aromatic group, saturated or partially saturated 5 or 6 membered heterocyclic having at least one ring nitrogen (see U.S. Patent 5,843,903).
• R-] 0 may be derived from an amino acid, having the structure - C(0)CH(NHRn)(R 12 ), in which Rn is H, or forms a C 3- membered alkylene with R ⁇ 2 .
• R ⁇ 2 may be H, alkyl, aminoalkyl, amino, hydroxyl, mercapto, phenyl, benzyl or methylthio (see U.S. Patent 4,296,105).
• exemplary anthracyclines are doxorubicin, daunorubicin, idarubicin, epirubicin, pirarubicin, zorubicin, and carubicin. Suitable compounds have the structures:
• Doxorubicin OCH; C(0)CH 2 OH OH out of ring plane
• Epirubicin (4' epimer of OCH ? C(0)CH 2 OH OH in ring plane doxorubicin)
• Daunorubicin OCH 3 C(0)CH 3 OH out of ring plane
• Idarubicin H C(0)CH 3 OH out of ring plane
• Carubicin OH C(0)CH 3 OH out of ring plane
• Other suitable anthracyclines are anthramycin, mitoxantrone, menogaril, nogalamycin, aclacinomycin A, olivomycin A, chromomycin A 3 , and plicamycin having the structures: t R, R,
• Olivomycin A COCH(CH 3 ) 2 CH, COCK, H Chromomycin A 3 COCH 3 CH, CCCR 3 CH, Plicamycin H H H CH,
• anthracyclines include, FCE 23762 doxorubicin derivative (Quaglia et al., J. Liq. Chromatogr. 17(18):3911-3923, 1994), annamycin (Zou et al., J. Pharm. Sci. 82(11 ):1151-1154, 1993), ruboxyl (Rapoport ef al., J. Controlled Release 58(2):153-162, 1999), anthracycline disaccharide doxorubicin analogue (Pratesi et al., Clin. Cancer Res.
• the ant-infective therapeutic agent is a fluoropyrimidine analog, such as 5-fluorouracil, or an analogue or derivative thereof, including carmofur, doxifluridine, emitefur, tegafur, and floxuridine.
• fluoropyrimidine analog such as 5-fluorouracil
• an analogue or derivative thereof including carmofur, doxifluridine, emitefur, tegafur, and floxuridine.
• Exemplary compounds have the structures:
• fluoropyrimidine analogues include 5-FudR (5- fluoro-deoxyuridine), or an analogue or derivative thereof, including 5- iododeoxyuridine (5-ludR), 5-bromodeoxyuridine (5-BudR), fluorouridine triphosphate (5-FUTP), and fluorodeoxyuridine monophosphate (5-dFUMP).
• 5-FudR 5- fluoro-deoxyuridine
• an analogue or derivative thereof including 5- iododeoxyuridine (5-ludR), 5-bromodeoxyuridine (5-BudR), fluorouridine triphosphate (5-FUTP), and fluorodeoxyuridine monophosphate (5-dFUMP).
• Exemplary compounds have the structures:
• fluoropyrimidine analogues include N3-alkylated analogues of 5-fluorouracil (Kozai et al., J. Chem. Soc, Perkin Trans.
• the anti-infective therapeutic agent is a folic acid antagonist, such as methotrexate or derivatives or analogues thereof, including edatrexate, trimetrexate, raltitrexed, piritrexim, denopterin, tomudex, and pteropterin.
• Methotrexate analogues have the following general structure:
• R group may be selected from organic groups, particularly those groups set forth in U.S. Patent Nos. 5,166,149 and 5,382,582.
• Ri may be N
• R 2 may be N or C(CH 3 )
• R 3 and R 3 ' may H or alkyl, e.g., CH 3
• R may be a single bond or NR, where R is H or alkyl group.
• R 5 , 6 , 8 may be H, OCH 3 , or alternately they can be halogens or hydro groups.
• R is a side chain of the general structure:
• the carboxyl groups in the side chain may be esterified or form a salt such as a Zn 2+ salt.
• R 9 and R 10 can be NH 2 or may be alkyl substituted.
• Exemplary folic acid antagonist compounds have the structures:
• cysteic acid and homocysteic acid methotrexate analogues (4,490,529), ⁇ -tert-butyl methotrexate esters (Rosowsky et al., J. Med. Chem. 28(5):660-7, 1985), fluorinated methotrexate analogues (Tsushima et al., Heterocycles 23(1 ):45-9, 1985), folate methotrexate analogue (Trombe, J. Bacteriol. 7-60(3):849-53, 1984), phosphonoglutamic acid analogues (Sturtz & Guillamot, Eur. J. Med. Chem.-Chim. Ther.
• the anti-infective therapeutic agent is a Podophyllotoxin, or a derivative or an analogue thereof.
• exemplary compounds of this type are etoposide or teniposide, which have the following structures:
• Other representative examples of podophyllotoxins include Cu(ll)- VP-16 (etoposide) complex (Tawa et al., Bioorg. Med. Chem. 6(7):1003-1008, 1998), pyrrolecarboxamidino-bearing etoposide analogues (Ji et al., Bioorg. Med. Chem. Lett.
• the anti-infective therapeutic agent is camptothecin, or an analogue or derivative thereof. Camptothecins have the following general structure.
• X is typically O, but can be other groups, e.g., NH in the case of 21-lactam derivatives.
• R-i is typically H or OH, but may be other groups, e.g., a terminally hydroxylated C ⁇ _ 3 alkane.
• R 2 is typically H or an ⁇ amino containing group such as (CH 3 ) 2 NHCH 2 , but may be other groups e.g., N0 2 , NH 2 , halogen (as disclosed in, e.g., U.S. Patent 5,552,156) or a short alkane containing these groups.
• R 3 is typically H or a short alkyl such as C 2 H 5 .
• R 4 is typically H but may be other groups, e.g., a methylenedioxy group with RL
• camptothecin compounds include topotecan, irinotecan (CPT-11), 9-aminocamptothecin, 21-lactam-20(S)-camptothecin, 10,11-methylenedioxycamptothecin, SN-38, 9-nitrocamptothecin, 10- hydroxycamptothecin.
• Exemplary compounds have the structures:
• Camptothecin H H H Topotecan: OH (CH 3 ) 2 NHCH 2 H SN-38: OH H C C 2 H 5 X: O for most analogs, NH for 21-lactam analogs
• Camptothecins have the five rings shown here.
• the ring labeled E must be intact (the lactone rather than carboxylate form) for maximum activity and minimum toxicity. Camptothecins are believed to function as topoisomerase I inhibitors and/or DNA cleavage agents.
• Hydroxyureas The anti-infective therapeutic agent of the present invention may be a hydroxyurea. Hydroxyureas have the following general structure:
• Suitable hydroxyureas are disclosed in, for example, U.S. Patent No. 6,080,874, wherein Ri is:
• R 2 is an alkyl group having 1-4 carbons and R 3 is one of H, acyl, methyl, ethyl, and mixtures thereof, such as a methylether.
• R-i is a cycloalkenyl group, for example N-(3-(5-(4- fluorophenylthio)-furyl)-2-cyclopenten-1-yl)N-hydroxyurea
• R 2 is H or an alkyl group having 1 to 4 carbons and R 3 is H
• X is H or a cation.
• Other suitable hydroxyureas are disclosed in, e.g., U.S.
• Other suitable hydroxyureas are disclosed in, e.g., U.S. Patent No. 5,066,658, wherein R 2 and R 3 together with the adjacent nitrogen form:
• hydroxyurea has the structure:
• the anti-infective therapeutic agent is a platinum compound.
• suitable platinum complexes may be of Pt(ll) or Pt(IV) and have this basic structure:
• X and Y are anionic leaving groups such as sulfate, phosphate, carboxylate, and halogen; Ri and R 2 are alkyl, amine, amino alkyl any may be further substituted, and are basically inert or bridging groups.
• Pt(ll) complexes Z-i and Z 2 are non-existent.
• Z-i and Z 2 may be anionic groups such as halogen, hydroxy, carboxylate, ester, sulfate or phosphate. See, e.g., U.S. Patent Nos. 4,588,831 and 4,250,189.
• Suitable platinum complexes may contain multiple Pt atoms. See, e.g., U.S. Patent Nos. 5,409,915 and 5,380,897.
• platinum compounds are cisplatin, carboplatin, oxaliplatin, and miboplatin having the structures:
• Oxaliplatin Other representative platinum compounds include (CPA) 2 Pt(DOLYM) and (DACH)Pt(DOLYM) cisplatin (Choi et al., Arch. Pharmacal Res. 22(2):151-156, 1999), Cis-(PtCI 2 (4,7-H-5-methyl-7- oxo)1 ,2,4(triazolo(1 ,5-a)pyrimidine) 2 ) (Navarro et al., J. Med. Chem. 41(3):332- 338, 1998), (Pt(cis-1 ,4-DACH)(trans-Cl 2 )(CBDCA)) .
• Dosages of Anti-Infective Agents The drug dose administered from the present compositions for prevention or inhibition of infection in accordance with the present invention will depend on a variety of factors, including the type of formulation, the location of the treatment site, and the type of condition being treated. However, certain principles can be applied in the application of this art. Drug dose can be calculated as a function of dose per unit area (of the treatment site), total drug dose administered can be measured and appropriate surface concentrations of active drug can be determined. Drugs are to be used at concentrations that range from several times more than to 50%, 20%, 10%, 5%, or even less than 1% of the concentration typically used in a single anti-infective systemic dose application.
• the anti- infective agent is released from the polymer composition in effective concentrations in a time period that may be measured from the time of infiltration into tissue adjacent to the device, which ranges from about less than 1 day to about 180 days. Generally, the release time may also be from about less than 1 day to about 180 days; from about 7 days to about 14 days; from about 14 days to about 28 days; from about 28 days to about 56 days; from about 56 days to about 90 days; from about 90 days to about 180 days.
• the exemplary anti-infective agents, used alone or in combination, should be administered under the following dosing guidelines.
• the total amount (dose) of anti-infective agent in the composition can be in the range of about 0.01 ⁇ g-1 ⁇ g, or about 1 ⁇ g-10 ⁇ g, or about 10 ⁇ g-1 mg, or about 1 mg to 10 mg, or about 10 mg-100 mg, or about 100 mg to 250 mg, or about 250 mg-1000 mg.
• the dose (amount) of anti-infective agent per unit area of device or tissue surface to which the agent is applied may be in the range of about 0.01 ⁇ g/mm 2 - 1 ⁇ g/mm 2 , or about 1 ⁇ g/mm 2 - 10 ⁇ g/mm 2 , or about 10 ⁇ g/mm 2 - 100 ⁇ g/mm 2 , or about 100 ⁇ g/mm 2 to 250 ⁇ g/mm 2 , or about 250 ⁇ g/mm 2 - 1000 ⁇ g/mm 2 .
• the above dosing parameters should be utilized in combination with the release rate of the drug from the composition such that a minimum concentration of about 10 ⁇ 8 M to 10 "7 M, or about 10 "7 M to 10 "6 M about 10 "6 M to 10 "5 M or about 10 "5 M to 10 "4 M of the agent is maintained on the tissue surface.
• the total dose of doxorubicin applied to the device or implant should not exceed 25 mg (range of 0.1 ⁇ g to 25 mg).
• the total amount of drug applied should be in the range of 1 ⁇ g to 5 mg.
• the dose per unit area i.e., the amount of drug as a function of the surface area of the portion of the implant to which drug is applied and/or incorporated
• doxorubicin should be applied to the implant surface at a dose of 0.1 ⁇ g/mm 2 - 10 ⁇ g/mm 2 .
• the above dosing parameters should be utilized in combination with the release rate of the drug from the implant surface such that a minimum concentration of 10 "7 - 10 "4 M of doxorubicin is maintained on the surface. It is necessary to insure that surface drug concentrations exceed concentrations of doxorubicin known to be lethal to multiple species of bacteria and fungi (i.e., are in excess of 10 ⁇ 4 M; although for some embodiments lower concentrations are sufficient).
• doxorubicin is released from the surface of the implant such that anti-infective activity is maintained for a period ranging from several hours to several months.
• the drug is released in effective concentrations for a period ranging from 1 week - 6 months.
• analogues and derivatives of doxorubicin (as described previously) with similar functional activity can be utilized for the purposes of this invention; the above dosing parameters are then adjusted according to the relative potency of the analogue or derivative as compared to the parent compound (e.g., a compound twice as potent as doxorubicin is administered at half the above parameters, a compound half as potent as doxorubicin is administered at twice the above parameters, etc.).
• the total dose of mitoxantrone applied should not exceed 5 mg (range of 0.01 ⁇ g to 5 mg). In a particularly preferred embodiment, the total amount of drug applied should be in the range of 0.1 ⁇ g to 1 mg.
• the dose per unit area i.e., the amount of drug as a function of the surface area of the portion of the implant to which drug is applied and/or incorporated should fall within the range of 0.01 ⁇ g - 20 ⁇ g per mm 2 of surface area.
• mitoxantrone should be applied to the implant surface at a dose of 0.05 ⁇ g/mm 2 - 3 ⁇ g/mm 2 .
• the above dosing parameters should be utilized in combination with the release rate of the drug from the implant surface such that a minimum concentration of 10 "5 - 10 "6 M of mitoxantrone is maintained. It is necessary to insure that drug concentrations on the implant surface exceed concentrations of mitoxantrone known to be lethal to multiple species of bacteria and fungi (i.e., are in excess of 10 s M; although for some embodiments lower drug levels will be sufficient).
• mitoxantrone is released from the surface of the implant such that anti-infective activity is maintained for a period ranging from several hours to several months.
• the drug is released in effective concentrations for a period ranging from 1 week - 6 months.
• analogues and derivatives of mitoxantrone (as described previously) with similar functional activity can be utilized for the purposes of this invention; the above dosing parameters are then adjusted according to the relative potency of the analogue or derivative as compared to the parent compound (e.g., a compound twice as potent as mitoxantrone is administered at half the above parameters, a compound half as potent as mitoxantrone is administered at twice the above parameters, etc.).
• the total dose of 5-fluorouracil applied should not exceed 250 mg (range of 1.0 ⁇ g to 250 mg). In a particularly preferred embodiment, the total amount of drug applied should be in the range of 10 ⁇ g to 25 mg.
• the dose per unit area i.e., the amount of drug as a function of the surface area of the portion of the implant to which drug is applied and/or incorporated
• 5-fluorouracil should be applied to the implant surface at a dose of 1.0 ⁇ g/mm 2 - 50 ⁇ g/mm 2 .
• the above dosing parameters should be utilized in combination with the release rate of the drug from the implant surface such that a minimum concentration of 10 "4 - 10 "7 M of 5-fluorouracil is maintained. It is necessary to insure that surface drug concentrations exceed concentrations of 5-fluorouracil known to be lethal to numerous species of bacteria and fungi (i.e., are in excess of 10 "4 M; although for some embodiments lower drug levels will be sufficient).
• 5-fluorouracil is released from the implant surface such that anti- infective activity is maintained for a period ranging from several hours to several months.
• the drug is released in effective concentrations for a period ranging from 1 week - 6 months.
• analogues and derivatives of 5-fluorouracil (as described previously) with similar functional activity can be utilized for the purposes of this invention; the above dosing parameters are then adjusted according to the relative potency of the analogue or derivative as compared to the parent compound (e.g., a compound twice as potent as 5-fluorouracil is administered at half the above parameters, a compound half as potent as 5-fluorouracil is administered at twice the above parameters, etc.).
• the total dose of etoposide applied should not exceed 25 mg (range of 0.1 ⁇ g to 25 mg). In a particularly preferred embodiment, the total amount of drug applied should be in the range of 1 ⁇ g to 5 mg.
• the dose per unit area i.e., the amount of drug as a function of the surface area of the portion of the implant to which drug is applied and/or incorporated should fall within the range of 0.01 ⁇ g - 100 ⁇ g per mm 2 of surface area.
• etoposide should be applied to the implant surface at a dose of 0.1 ⁇ g/mm 2 - 10 ⁇ g/mm 2 .
• the above dosing parameters should be utilized in combination with the release rate of the drug from the implant surface such that a concentration of 10 "5 - 10 "6 M of etoposide is maintained. It is necessary to insure that surface drug concentrations exceed concentrations of etoposide known to be lethal to a variety of bacteria and fungi (i.e., are in excess of 10 "5 M; although for some embodiments lower drug levels will be sufficient).
• etoposide is released from the surface of the implant such that anti-infective activity is maintained for a period ranging from several hours to several months.
• the drug is released in effective concentrations for a period ranging from 1 week - 6 months.
• analogues and derivatives of etoposide (as described previously) with similar functional activity can be utilized for the purposes of this invention; the above dosing parameters are then adjusted according to the relative potency of the analogue or derivative as compared to the parent compound (e.g., a compound twice as potent as etoposide is administered at half the above parameters, a compound half as potent as etoposide is administered at twice the above parameters, etc.).
• anthracyclines e.g., doxorubicin or mitoxantrone
• fluoropyrimidines e.g., 5-fluorouracil
• folic acid antagonists e.g., methotrexate and/or podophylotoxins (e.g., etoposide)
• podophylotoxins e.g., etoposide
• anthracyclines e.g., doxorubicin or mitoxantrone
• fluoropyrimidines e.g., 5-fluorouracil
• folic acid antagonists e.g., methotrexate and/or podophylotoxins (e.g., etoposide) may be utilized to enhance the antibacterial activity of the composition.
• compositions may further include a compound which acts to have an inhibitory effect on pathological processes in or around the treatment site.
• additional therapeutically active agents include, by way of example and not limitation, anti-thrombotic agents, anti-proliferative agents, anti-inflammatory agents, neoplastic agents, enzymes, receptor antagonists or agonists, hormones, antibiotics, antimicrobial agents, antibodies, cytokine inhibitors, IMPDH (inosine monophosplate dehydrogenase) inhibitors tyrosine kinase inhibitors, MMP inhibitors, p38 MAP kinase inhibitors, immunosuppressants, apoptosis antagonists, caspase inhibitors, and JNK inhibitor.
• anti-thrombotic agents include, by way of example and not limitation, anti-thrombotic agents, anti-proliferative agents, anti- inflammatory agents, neoplastic agents, enzymes, receptor antagonists or agonists, hormones, antibiotics, antimicrobial agents, antibodies, cytokine inhibitors, IMPDH (inosine monophosplate dehydrogenase) inhibitors tyrosine kin
• the polymeric composition may further include an anti-thrombotic agent and/or antiplatelet agent and/or a thrombolytic agent, which reduces the likelihood of thrombotic events upon implantation of a medical implant.
• anti-thrombotic and/or antiplatelet and/or thrombolytic agents include heparin, heparin fragments, organic salts of heparin, heparin complexes (e.g., benzalkonium heparinate, tridodecylammonium heparinate), dextran, sulfonated carbohydrates such as dextran sulfate, coumadin, coumarin, heparinoid, danaparoid, argatroban chitosan sulfate, chondroitin sulfate, danaparoid, lepirudin, hirudin, AMP, adenosine, 2-chloroadenosine, acetylsalicylic acid, phenylbutazone
• Further examples include plasminogen, lys- plasminogen, alpha-2-antiplasmin, urokinase, aminocaproic acid, ticlopidine, clopidogrel, trapidil (triazolopyrimidine), naftidrofuryl, auriritricarboxylic acid and glycoprotein llb/llla inhibitors such as abcixamab, eptifibatide, and tirogiban.
• agents capable of affecting the rate of clotting include glycosaminoglycans, danaparoid, 4-hydroxycourmarin, warfarin sodium, dicumarol, phenprocoumon, indan-1 ,3-dione, acenocoumarol, anisindione, and rodenticides including bromadiolone, brodifacoum, diphenadione, chlorophacinone, and pidnone.
• the polymeric formulation may be or include a hydrophilic polymer gel that itself has anti-thrombogenic properties.
• the composition can be in the form of a coating that can comprise a hydrophilic, biodegradable polymer that is physically removed from the surface of the device over time, thus reducing adhesion of platelets to the device surface.
• the gel composition can include a polymer or a blend of polymers.
• Representative examples include alginates, chitosan and chitosan sulfate, hyaluronic acid, dextran sulfate, PLURONIC polymers (e.g., F-127 or F87), chain extended PLURONIC polymers, various polyester-polyether block copolymers of various configurations (e.g., AB, ABA, or BAB, where A is a polyester such as PLA, PGA, PLGA, PCL or the like), examples of which include MePEG-PLA, PLA-PEG-PLA, and the like).
• PLURONIC polymers e.g., F-127 or F87
• chain extended PLURONIC polymers e.g., various polyester-polyether block copolymers of various configurations (e.g., AB, ABA, or BAB, where A is a polyester such as PLA, PGA, PLGA, PCL or the like), examples of which include MePEG-PLA, PLA-PEG-PL
• the anti-thrombotic composition can include a crosslinked gel formed from a combination of molecules (e.g., PEG) having two or more terminal electrophilic groups and two or more nucleophilic groups.
• the polymeric formulation may further include an agent from one of the following classes of compounds: anti-inflammatory agents (e.g., dexamethasone, cortisone, fludrocortisone, prednisone, prednisolone, 6 ⁇ - methylprednisolone, triamcinolone, betamethasone, and aspirin); MMP inhibitors (e.g., batimistat, marimistat, TIMP's representative examples of which are included in U.S. Patent Nos. 5,665,777; 5,985,911 ; 6,288,261 ; 5,952,320;
• anti-inflammatory agents e.g., dexamethasone, cortisone, fludrocortisone, prednisone, prednisolone, 6 ⁇ - methylpred
• WO 00/63204A2 WO 01/21591 A1 , WO 01/35959A1 , WO 01/74811A2, WO 02/18379A2, WO 02/064594A2, WO 02/083622A2, WO 02/094842A2,WO 02/096426A1 , WO 02/101015A2, WO 02/103000A2, WO 03/008413A1 , WO 03/016248A2, WO 03/020715A1 , WO 03/024899A2, WO 03/031431 A1 , WO 03/040103A1 , WO 03/053940A1 , WO 03/053941 A2, WO 03/063799A2, WO 03/079986A2, WO 03/080024A2, WO 03/082287A1 , WO 97/44467A1 , WO 99/01449A1 , and WO 99/58523A1 ), and immunomodulatory
• Patent No. 6,258,823 and everolimus and derivatives thereof (e.g., U.S. Patent No. 5,665,772).
• Further representative examples of sirolimus analogues and derivatives include ABT-578 and those found in PCT Publication Nos.
• biologically active agents which may be included in the compositions of the invention include tyrosine kinase inhibitors, such as imantinib, ZK-222584, CGP-52411 , CGP-53716, NVP-AAK980-NX, CP-127374, CP-564959, PD-171026, PD-173956, PD-180970, SU-0879, and SKI-606; MMP inhibitors such as nimesulide, PKF-241-466, PKF-242-484, CGS-27023A, SAR-943, primomastat, SC-77964, PNU-171829, AG-3433, PNU-142769, SU-5402, and Dexlipotam; p38 MAP kinase inhibitors such as include CGH-2466 and PD-98-59; immunosuppressants such as argyrin B, macrocyclic lactone, ADZ-62-826, CCI-779, tilomisole,
• the composition may further include an antibiotic (e.g., amoxicillin, trimethoprim-sulfamethoxazole, azithromycin, clarithromycin, amoxicillin-clavulanate, cefprozil, cefuroxime, cefpodoxime, or cefdinir).
• an antibiotic e.g., amoxicillin, trimethoprim-sulfamethoxazole, azithromycin, clarithromycin, amoxicillin-clavulanate, cefprozil, cefuroxime, cefpodoxime, or cefdinir.
• a polymeric composition comprising a fibrosis- inhibiting agent is combined with an agent that can modify metabolism of the agent in vivo to enhance efficacy of the fibrosis-inhibiting agent.
• One class of therapeutic agents that can be used to alter drug metabolism includes agents capable of inhibiting oxidation of the anti-scarring agent by cytochrome P450 (CYP).
• compositions include a fibrosis- inhibiting agent (e.g., paclitaxel, rapamycin, everolimus) and a CYP inhibitor, which may be combined (e.g., coated) with any of the devices described herein, including, without limitation, stents, grafts, patches, valves, wraps, and films.
• a fibrosis- inhibiting agent e.g., paclitaxel, rapamycin, everolimus
• a CYP inhibitor include flavones, azole antifungals, macrolide antibiotics, HIV protease inhibitors, and anti-sense oligomers.
• Devices comprising a combination of a fibrosis-inhibiting agent and a CYP inhibitor may be used to treat a variety of proliferative conditions that can lead to undesired scarring of tissue, including intimal hyperplasia, surgical adhesions, and tumor growth.
• a polymeric composition comprising an anti- infective agent (e.g., anthracyclines (e.g., doxorubicin or mitoxantrone), fluoropyrimidines (e.g., 5-fluorouracil), folic acid antagonists (e.g., methotrexate and/or podophylotoxins (e.g., etoposide)) can be combined with traditional antibiotic and/or antifungal agents to enhance efficacy.
• an anti- infective agent e.g., anthracyclines (e.g., doxorubicin or mitoxantrone), fluoropyrimidines (e.g., 5-fluorouracil), folic acid antagonists (e.g., methotrexate and/or podophylotoxins (e.g., etoposide)
• an anti- infective agent e.g., anthracyclines (e.g., doxorubi
• the anti-infective agent may be further combined with anti-thrombotic and/or antiplatelet agents (for example, heparin, dextran sulfate, danaparoid, lepirudin, hirudin, AMP, adenosine, 2-chloroadenosine, aspirin, phenylbutazone, indomethacin, meclofenamate, hydrochloroquine, dipyridamole, iloprost, ticlopidine, clopidogrel, abcixamab, eptifibatide, tirofiban, streptokinase, and/or tissue plasminogen activator) to enhance efficacy.
• anti-thrombotic and/or antiplatelet agents for example, heparin, dextran sulfate, danaparoid, lepirudin, hirudin, AMP, adenosine, 2-chloroadenosine, aspirin, phenylbut
• paclitaxel should be understood to refer to not only the common chemically available form of paclitaxel, but analogues (e.g., TAXOTERE, as noted above) and paclitaxel conjugates (e.g., paclitaxel-PEG, paclitaxel-dextran, or paclitaxel-xylos).
• analogues e.g., TAXOTERE, as noted above
• paclitaxel conjugates e.g., paclitaxel-PEG, paclitaxel-dextran, or paclitaxel-xylos.
• agents set forth above may be noted within the context of one class, many of the agents listed in fact have multiple biological activities. Further, more than one therapeutic agent may be utilized at a time (i.e., in combination), or delivered sequentially.
• compositions and Methods for Generating Compositions Which Comprise a Therapeutic Agent The present invention provides various compositions which can be used to inhibit fibrosis and/or infection of tissue in the vicinity of a treatment site (e.g., a surgical site).
• a treatment site e.g., a surgical site.
• fibrosis and/or infection is inhibited by local or systemic release of specific pharmacological agents that become localized at the site or intervention.
• fibrosis and/or infection can be inhibited by local or systemic release of specific pharmacological agents that become localized adjacent to a device or implant that has been introduced into a host.
• compositions are provided which inhibit fibrosis in and around an implanted device, or prevent "stenosis" of a device/implant in situ, thus enhancing the efficacy.
• anti-infective compositions are provided which inhibit or prevent infection in and around an implanted device.
• the therapeutic agent may be administered as a sustained low dose therapy to prevent disease progression, prolong disease remission, or decrease symptoms in active disease.
• the therapeutic agent may be administered in higher doses as a "pulse" therapy to induce remission in acutely active disease.
• the minimum dose capable of achieving these endpoints can be used and can vary according to patient, severity of disease, formulation of the administered agent, potency and tolerability of the therapeutic agent, and route of administration.
• several techniques would be suitable to achieve preferentially elevated levels of therapeutic agents in the vicinity of the area to be treated.
• fibrosis-inhibiting agents include: (a) using drug-delivery catheters and/or a syringe and needle for local, regional or systemic delivery of fibrosis-inhibiting agents to the tissue surrounding the device or implant (typically, drug delivery catheters are advanced through the circulation or inserted directly into tissues under radiological guidance until they reach the desired anatomical location; the fibrosis-inhibiting agent can then be released from the catheter lumen in high local concentrations in order to deliver therapeutic doses of the drug to the tissue surrounding the device or implant); (b) drug localization techniques such as magnetic, ultrasonic or MRI-guided drug delivery; (c) chemical modification of the therapeutic drug or formulation designed to increase uptake of the agent into damaged tissues (e.g., antibodies directed against damaged or healing tissue components such as macrophages, neutrophils, smooth muscle cells, fibroblasts, extracellular matrix components, neovascular tissue); (d) chemical modification of the therapeutic drug or formulation designed to localize the drug to areas of bleeding or disrupted vascul
• the tissue cavity or surgical pocket into which a device or implant is placed can be treated with an anti-infective and/or fibrosis- inhibiting therapeutic agent prior to, during, or after the procedure.
• This can be accomplished in several ways including: (a) topical application of the agent into the anatomical space or surface where the device will be placed (particularly useful for this embodiment is the use of polymeric carriers which release the agent over a period ranging from several hours to several weeks.
• compositions that can be used for this application include, e.g., fluids, microspheres, pastes, gels, microparticulates, sprays, aerosols, solid implants and other formulations which release a therapeutic agent into the region where the device or implant will be implanted); (b) microparticulate forms of the therapeutic agent are also useful for directed delivery into the implantation site; (c) sprayable collagen-containing formulations such as COSTASIS and crosslinked derivatized poly(ethylene glycol) -collagen compositions (described, e.g., in U.S. Patent Nos.
• CT3 both from Angiotech Pharmaceuticals, Inc., Canada
• sprayable PEG-containing formulations such as COSEAL or ADHIBIT (Angiotech Pharmaceuticals, Inc.), SPRAYGEL or DURASEAL (both from Confluent Surgical, Inc., Boston, MA), either alone, or loaded with a therapeutic agent, applied to the implantation site (or the implant/device surface
• fibrin-containing formulations such as FLOSEAL or TISSEEL (both from Baxter Healthcare Corporation, Fremont, CA), applied to the implantation site (or the implant/device surface
• hyaluronic acid- containing formulations such as RESTYLANE or PERLANE (both from Q-Med AB, Sweden), HYLAFORM (Inamed Corporation (Santa Barbara, CA)), SYNVISC (Bio
• DERMABOND Johnson & Johnson, Inc., New Brunswick, NJ
• INDERMIL U.S. Surgical Company, Norwalk, CT
• GLUSTITCH Blacklock Medical Products Inc., Canada
• TISSUMEND II Veterniary Products Laboratories,, Phoenix, AZ
• VETBOND 3M Company, St. Paul, MN
• HISTOACRYL BLUE Davis & Geek, St.
• a preferred polymeric matrix which can be used to help prevent the formation of fibrous tissue, either alone or in combination with a fibrosis inhibiting agent/composition is formed from reactants comprising either one or both of pentaerythritol poly(ethy!ene g!ycol)ether tetra-sulfhydryl] (4-armed thiol PEG, which includes structures having a linking group(s) between a sulfhydryl group(s) and the terminus of the polyethylene glycol backbone) and pentaerythritol poly(ethylene glycol)ether tetra-succinimidyl glutarate] (4-armed NHS PEG, which again includes structures having a linking group(s) between a NHS group(s) and the terminus of the polyethylene glycol backbone) as reactive reagents.
• reactants comprising either one or both of pentaerythritol poly(ethy!ene g!ycol)ether tetra-s
• Another preferred composition comprises either one or both of pentaerythritol poly(ethylene glycol)ether tetra-amino] (4-armed amino PEG, which includes structures having a linking group(s) between an amino group(s) and the terminus of the polyethylene glycol backbone) and pentaerythritol poly(ethylene glycol)ether tetra-succinimidyl glutarate] (4-armed NHS PEG, which again includes structures having a linking group(s) between a NHS group(s) and the terminus of the polyethylene glycol backbone) as reactive reagents.
• Chemical structures for these reactants are shown in, e.g., U.S. Patent 5,874,500.
• collagen or a collagen derivative is added to the poly(ethylene glycol)-containing reactant(s) to form a preferred crosslinked matrix that can serve as a polymeric carrier for a therapeutic agent or a stand-alone composition to help prevent the formation of fibrous tissue.
• collagen or a collagen derivative e.g., methylated collagen
• desired therapeutic agents may be admixed with, blended with, conjugated to, or, otherwise modified to contain a polymer composition (which may be either biodegradable or non- biodegradable) or a non-polymeric composition in order to release the therapeutic agent over a prolonged period of time.
• a polymer composition which may be either biodegradable or non- biodegradable
• a non-polymeric composition in order to release the therapeutic agent over a prolonged period of time.
• localized delivery as well as localized sustained delivery of the fibrosis-inhibiting and/or anti-infective agent may be required.
• a desired therapeutic agent may be admixed with, blended with, conjugated to, or, otherwise modified to contain a polymeric composition (which may be either biodegradable or non-biodegradable) or non-polymeric composition in order to release the therapeutic agent over a period of time.
• a polymeric composition which may be either biodegradable or non-biodegradable
• non-polymeric composition in order to release the therapeutic agent over a period of time.
• biodegradable polymers suitable for the delivery of the aforementioned therapeutic agents include albumin, collagen, gelatin, hyaluronic acid, starch, cellulose and cellulose derivatives (e.g., regenerated cellulose, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, cellulose acetate phthalate, cellulose acetate succinate, hydroxypropylmethylcellulose phthalate), casein, dextrans, polysaccharides, fibrinogen, poly( ether ester) multiblock copolymers, based on poly(ethylene glycol) and poly(butylene terephthalate), tyrosine-derived polycarbonates (e.g., U.S. Patent No.
• X is
• non-degradable polymers suitable for the delivery of the aforementioned therapeutic agents include poly(ethylene-co- vinyl acetate) ("EVA") copolymers, aromatic polyesters, such as poly(ethylene terephthalate), silicone rubber, acrylic polymers (polyacrylate, polyacrylic acid, polymethylacrylic acid, polymethylmethacrylate, poly(butyl methacrylate)), poly(alkylcynoacrylate) (e.g., poly(ethylcyanoacrylate), poly(butylcyanoacrylate) poly(hexylcyanoacrylate) poly(octylcyanoacrylate)), acrylic resin, polyethylene, polypropylene, polyamides (nylon 6,6), polyurethanes (e.g., CHRONOFLEX AL and CHRONOFLEX AR (both from CardioTech International, Inc., Woburn, MA), TECOFLEX, and BIONATE (Polymer Technology Group, Inc., Emeryville, CA)),, poly(ester (EVA
• polystyrene poly(styrene sulfonic acid), poly(styrene)-block-poly(isobutylene)- block-poly(styrene), poly(styrene)-poly(isoprene) block copolymers
• vinyl polymers polyvinylpyrrolidone, poly(vinyl alcohol), poly(vinyl acetate phthalate) as well as copolymers and blends thereof.
• Polymers may also be developed which are either anionic (e.g., alginate, carrageenan, carboxymethyl cellulose, poly(acrylamido-2-methyl propane sulfonic acid) and copolymers thereof, poly(methacrylic acid and copolymers thereof and poly(acrylic acid) and copolymers thereof, as well as blends thereof, or cationic (e.g., chitosan, poly- L-lysine, polyethylenimine, and poly(allyl amine)) and blends thereof (see generally, Dunn et al., J. Applied Polymer Sci. 50:353-365, 1993; Cascone et al., J.
• anionic e.g., alginate, carrageenan, carboxymethyl cellulose, poly(acrylamido-2-methyl propane sulfonic acid) and copolymers thereof, poly(methacrylic acid and copolymers thereof and poly(acrylic acid) and copolymers thereof, as well as
• polysaccharides such as hyaluronic acid, chitosan and fucans, and copolymers of polysaccharides with degradable polymers.
• Other representative polymers capable of sustained localized delivery of anti-infective and/or fibrosis-inhibiting therapeutic agents include carboxylic polymers, polyacetates, polycarbonates, polyethers, polyethylenes, polyvinylbutyrals, polysilanes, polyureas, polyoxides, polystyrenes, polysulfides, polysulfones, polysulfonides, polyvinylhalides, pyrrolidones, rubbers, thermal- setting polymers, cross-linkable acrylic and methacrylic polymers, ethylene acrylic acid copolymers, styrene acrylic copolymers, vinyl acetate polymers and copolymers, vinyl acetal polymers and copolymers, epoxies
• Patent Application Publication Nos. 2003/0068377, 2002/0192286, 2002/0076441 , and 2002/0090398 can also be blended or copolymerized in various compositions as required to deliver therapeutic doses of biologically active agents.
• Polymeric carriers for anti-infective and/or fibrosis-inhibiting therapeutic agents can be fashioned in a variety of forms, with desired release characteristics and/or with specific properties depending upon the composition being utilized.
• polymeric carriers may be fashioned to release a therapeutic agent upon exposure to a specific triggering event such as pH (see, e.g., Heller et al., "Chemically Self-Regulated Drug Delivery Systems," in Polymers in Medicine III, Elsevier Science Publishers B.V., Amsterdam, 1988, pp. 175-188; Kang et al., J. Applied Polymer Sci. 48:343-354, 1993; Dong et al., J. Controlled Release 19:171-178, 1992; Dong and Hoffman, J. Controlled Release 75:141-152, 1991 ; Kim et al., J. Controlled Release 28:143-152, 1994; Comejo-Bravo et al., J.
• a specific triggering event such as pH (see, e.g., Heller et al., "Chemically Self-Regulated Drug Delivery Systems," in Polymers in Medicine III, Elsevier Science Publishers B.V., Amsterdam, 1988, pp. 175
• pH-sensitive polymers include poly (acrylic acid) and its derivatives (including for example, homopolymers such as poly(aminocarboxylic acid); poly(acrylic acid); poly(methyl acrylic acid), copolymers of such homopolymers, and copolymers of poly(acrylic acid) and/or acrylate or acrylamide Imonomers such as those discussed above.
• Other pH sensitive polymers include polysaccharides such as cellulose acetate phthalate; hydroxypropylmethylcellulose phthalate; hydroxypropylmethylcellulose acetate succinate; cellulose acetate trimellilate; and chitosan.
• Yet other pH sensitive polymers include any mixture of a pH sensitive polymer and a water-soluble polymer.
• ant-infective and/or fibrosis-inhibiting therapeutic agents can be delivered via polymeric carriers which are temperature sensitive (see, e.g., Chen et al., "Novel Hydrogels of a Temperature-Sensitive PLURONIC Grafted to a Bioadhesive Polyacrylic Acid Backbone for Vaginal Drug Delivery," in Proceed. Intern. Symp. Control. Rel. Bioact. Mater. 22:167-168, Controlled Release Society, Inc., 1995; Okano, "Molecular Design of Stimuli-Responsive Hydrogels for Temporal Controlled Drug Delivery," in Proceed. Intern. Symp. Control. Rel. Bioact. Mater.
• thermogelling polymers and the gelatin temperature (LCST (°C)
• LCST gelatin temperature
• homopolymers such as poly(N-methyl-N-n-propylacrylamide), 19.8; poly(N-n-propylacrylamide), 21.5; poly(N-methyl-N-isopropylacrylamide), 22.3; poly(N-n-propylmethacrylamide), 28.0; poly(N-isopropylacrylamide), 30.9; poly(N, n-diethylacrylamide), 32.0; poly(N-isopropylmethacrylamide), 44.0; poly(N-cyclopropylacrylamide), 45.5; poly(N-ethylmethyacrylamide), 50.0; poly(N-methyl-N-ethylacrylamide), 56.0; poly(N-cyclopropylmethacrylamide), 59.0; poly(N-ethylacrylamide), 72.0.
• thermogelling polymers may be made by preparing copolymers between (among) monomers of the above, or by combining such homopolymers with other water-soluble polymers such as acrylmonomers (e.g., acrylic acid and derivatives thereof, such as methylacrylic acid, acrylate monomers and derivatives thereof, such as butyl methacrylate, butyl acrylate, lauryl acrylate, and acrylamide monomers and derivatives thereof, such as N-butyl acrylamide and acrylamide).
• acrylmonomers e.g., acrylic acid and derivatives thereof, such as methylacrylic acid, acrylate monomers and derivatives thereof, such as butyl methacrylate, butyl acrylate, lauryl acrylate, and acrylamide monomers and derivatives thereof, such as N-butyl acrylamide and acrylamide.
• thermogelling polymers include cellulose ether derivatives such as hydroxypropyl cellulose, 41 °C; methyl cellulose, 55°C; hydroxypropylmethyl cellulose, 66°C; and ethylhydroxyethyl cellulose, polyalkylene oxide-polyester block copolymers of the structure X-Y, Y-X-Y and X-Y-X where X in a polyalkylene oxide and Y is a biodegradable polyester (e.g., PLG-PEG-PLG) and PLURONICs such as F-127, 10 - 15°C; L-122, 19°C; L-92, 26°C; L-81 , 20°C; and L-61 , 24°C.
• PLG-PEG-PLG biodegradable polyester
• PLURONICs such as F-127, 10 - 15°C; L-122, 19°C; L-92, 26°C; L-81 , 20°C; and L-61
• patents relating to thermally gelling polymers and the preparation include U.S. Patent Nos. 6,451 ,346; 6,201 ,072; 6,117,949; 6,004,573; 5,702,717; and 5,484,610; and PCT Publication Nos. WO 99/07343; WO 99/18142; WO 03/17972; WO 01/82970; WO 00/18821 ; WO 97/15287; WO 01/41735; WO 00/00222 and WO 00/38651.
• Anti-infective and/or fibrosis-inhibiting therapeutic agents may be linked by occlusion in the polymer, dissolution in the polymer, bound by covalent linkages, bound by ionic interactions, or encapsulated in microcapsules.
• therapeutic compositions are provided in non-capsular formulations such as microspheres (ranging from nanometers to micrometers in size), pastes, threads of various size, films, or sprays.
• the anti-scarring agent may be incorporated into biodegradable magnetic nanospheres.
• the nanospheres may be used, for example, to replenish an anti-scarring agent into an implanted intravascular device, such as a stent containing a weak magnetic alloy (see, e.g., Z. Forbes, B. B. Yellen, G. Friedman, K. Barbee. "An approach to targeted drug delivery based on uniform magnetic fields," IEEE Trans. Magn. 39(5): 3372-3377 (2003)).
• an implanted intravascular device such as a stent containing a weak magnetic alloy
• therapeutic compositions of anti-infective and/or fibrosis-inhibiting agents may be fashioned in the form of microspheres, microparticles and/or nanoparticles having any size ranging from 50 nm to 500 ⁇ m, depending upon the particular use. These compositions can be.
• compositions can be formed by spray-drying methods, milling methods, coacervation methods, W/O emulsion methods, W/O/W emulsion methods, and solvent evaporation methods.
• these compositions can include microemulsions, emulsions, liposomes and micelles.
• such compositions may also be readily applied as a "spray", which solidifies into a film or coating for use as a device/implant surface coating or to line the tissues of the implantation site.
• Such sprays may be prepared from microspheres of a wide array of sizes, including for example, from 0.1 ⁇ m to 3 ⁇ m, from 10 ⁇ m to 30 ⁇ m, and from 30 ⁇ m to 100 ⁇ m.
• compositions that include anti-infective and/or anti- fibrosis agents may also be prepared in a variety of "paste" or gel forms.
• therapeutic compositions are provided which are liquid at one temperature (e.g., temperature greater than 37°C, such as 40°C, 45°C, 50°C, 55°C or 60°C), and solid or semi-solid at another temperature (e.g., ambient body temperature, or any temperature lower than 37°C).
• temperature e.g., temperature greater than 37°C, such as 40°C, 45°C, 50°C, 55°C or 60°C
• solid or semi-solid e.g., ambient body temperature, or any temperature lower than 37°C.
• Such "thermopastes” may be readily made utilizing a variety of techniques (see, e.g., PCT Publication WO 98/24427).
• pastes may be applied as a liquid, which solidify in vivo due to dissolution of a water-soluble component of the paste and precipitation of encapsulated drug into the aqueous body environment.
• These "pastes” and “gels” containing therapeutic agents are particularly useful for application to the surface of tissues that will be in contact with the implant or device.
• polymeric carriers are provided which are adapted to contain and release a hydrophobic ant- infective and/or fibrosis-inhibiting compound, and/or the carrier containing the hydrophobic compound in combination with a carbohydrate, protein or polypeptide.
• the polymeric carrier contains or comprises regions, pockets, or granules of one or more hydrophobic compounds.
• hydrophobic compounds may be incorporated within a matrix which contains the hydrophobic therapeutic compound, followed by incorporation of the matrix within the polymeric carrier.
• matrices can be utilized in this regard, including for example, carbohydrates and polysaccharides such as starch, cellulose, dextran, methylcellulose, sodium alginate, heparin, chitosan and hyaluronic acid, proteins or polypeptides such as albumin, collagen and gelatin.
• hydrophobic compounds may be contained within' a hydrophobic core, and this core contained within a hydrophilic shell.
• the anti-infective and/or fibrosis-inhibiting therapeutic agent may be delivered as a solution.
• the therapeutic agent can be incorporated directly into the solution to provide a homogeneous solution or dispersion.
• the solution is an aqueous solution.
• the aqueous solution may further include buffer salts, as well as viscosity modifying agents (e.g., hyaluronic acid, alginates, carboxymethylcellulose (CMC), and the like).
• the solution can include a biocompatible solvent or liquid oligomers and/or polymers, such as ethanol, DMSO, glycerol, PEG-200, PEG-300 or NMP.
• compositions may further comprise a polymer such a degradable polyester, where the polyester may comprise the residues of one or more of the monomers selected from lactide, lactic acid, glycolide, glycolic acid, e-caprolactone, gamma-caprolactone, hydroxyvaleric acid, hydroxybutyric acid, beta-butyrolactone, gamma-butyrolactone, gamma- valerolactone, ⁇ -decanolactone, ⁇ -decanolactone, trimethylene carbonate, 1 ,4- dioxane-2-one or 1 ,5-dioxepan-2one, or block copolymers of the form X-Y, Y-X- Y, R-(Y-X) n - R-(X-Y)n and X-Y-X (where X in a polyalkylene oxide (e.g., poly(ethylene glycol, poly(propylene glycol) and block copolymers of poly(ethylene
• the therapeutic anti- infective and/or fibrosis-inhibiting agent can further comprise a secondary carrier.
• the secondary carrier can be in the form of microspheres (e.g., PLGA, PLLA, PDLLA, PCL, gelatin, polydioxanone, poly(alkylcyanoacrylate)), nanospheres (PLGA, PLLA, PDLLA, PCL, gelatin, polydioxanone, poly(alkylcyanoacrylate)), liposomes, emulsions, microemulsions, micelles (SDS, block copolymers of the form X-Y, Y-X-Y, R-(Y-X) n , R-(X-Y) n and X-Y-X (where X in a polyalkylene oxide (e.g., poly(ethylene glycol, poly(propylene glycol) and block copolymers of poly(ethylene oxide) and poly(propylene oxide) (e.g., X
• hydroxypropyl cyclodextrin (Cserhati and Hollo, Int. J. Pharm. 708:69- 75, 1994), liposomes (see, e.g., Sharma et al., Cancer Res. 53:5877-5881 , 1993; Sharma and Straubinger, Pharm. Res. 77(60):889-896, 1994; WO 93/18751 ; U.S. Patent No. 5,242,073), liposome/gel (WO 94/26254), nanocapsules (Bartoli et al., J.
• polymeric carriers can be materials that are formed in situ.
• the precursors can be monomers or macromers that contain unsaturated groups that can be polymerized and/or cross-linked.
• the monomers or macromers can then, for example, be injected into the treatment area or onto the surface of the treatment area and polymerized in situ using a radiation source (e.g., visible or UV light) or a free radical system (e.g., potassium persulfate and ascorbic acid or iron and hydrogen peroxide).
• a radiation source e.g., visible or UV light
• a free radical system e.g., potassium persulfate and ascorbic acid or iron and hydrogen peroxide.
• the polymerization step can be performed immediately prior to, simultaneously to or post injection of the reagents into the treatment site.
• Representative examples of compositions that undergo free radical polymerization reactions are described in WO 01/44307, WO 01/68720, WO 02/072166, WO 03/043552, WO 93/17669, WO 00/64977; U.S. Patent Nos.
• compositions that can be administered as liquids, but subsequently form hydrogels at the site of administration.
• Such in situ hydrogel forming compositions can be administered as liquids from a variety of different devices, and are more adaptable for administration to any site, since they are not preformed.
• examples of in situ forming hydrogels include photoactivatable mixtures of water-soluble co-polyester prepolymers and polyethylene glycol to create hydrogel barriers.
• Block copolymers of polyalkylene oxide polymers e.g., PLURONIC compounds from BASF Corporation, Mount Olive, NJ
• poloxamers have been designed that are soluble in cold water, but form insoluble hydrogels that adhere to tissues at body temperature (Leach, et al., Am. J. Obstet. Gynecol. 162:1317-1319 (1990)).
• the present invention provides for polymeric crosslinked matrices, and polymeric carriers, that may be used to assist in the prevention of the formation or growth of fibrous connective tissue.
• the composition may contain and deliver fibrosis-inhibiting agents in the vicinity of the implanted device.
• the following compositions are particularly useful when it is desired to infiltrate around the device, with or without a fibrosis- inhibiting agent.
• Such polymeric materials may be prepared from, e.g., (a) synthetic materials, (b) naturally-occurring materials, or (c) mixtures of synthetic and naturally occurring materials.
• the matrix may be prepared from, e.g., (a) a one-component, i.e., self-reactive, compound, or (b) two or more compounds that are reactive with one another.
• these materials are fluid prior to delivery, and thus can be sprayed or otherwise extruded from a delivery device (e.g., a syringe) in order to deliver the composition.
• a delivery device e.g., a syringe
• the component materials react with each other, and/or with the body, to provide the desired affect.
• materials that are reactive with one another must be kept separated prior to delivery to the patient, and are mixed together just prior to being delivered to the patient, in order that they maintain a fluid form prior to delivery.
• the components of the matrix are delivered in a liquid state to the desired site in the body, whereupon in situ polymerization occurs.
• crosslinked polymer compositions are prepared by reacting a first synthetic polymer containing two or more nucleophilic groups with a second synthetic polymer containing two or more electrophilic groups, where the electrophilic groups are capable of covalently binding with the nucleophilic groups.
• the first and second polymers are each non-immunogenic.
• the matrices are not susceptible to enzymatic cleavage by, e.g., a matrix metalloproteinase (e.g., collagenase) and are therefore expected to have greater long-term persistence in vivo than collagen-based compositions.
• polymer refers inter alia to polyalkyls, polyamino acids, polyalkyleneoxides and polysaccharides. Additionally, for external or oral use, the polymer may be polyacrylic acid or carbopol.
• synthetic polymer refers to polymers that are not naturally occurring and that are produced via chemical synthesis. As such, naturally occurring proteins such as collagen and naturally occurring polysaccharides such as hyaluronic acid are specifically excluded. Synthetic collagen, and synthetic hyaluronic acid, and their derivatives, are included.
• Multifunctionally activated synthetic polymers Synthetic polymers containing either nucleophilic or electrophilic groups are also referred to herein as "multifunctionally activated synthetic polymers.”
• multifunctionally activated refers to synthetic polymers which have, or have been chemically modified to have, two or more nucleophilic or electrophilic groups which are capable of reacting with one another (i.e., the nucleophilic groups react with the electrophilic groups) to form covalent bonds.
• Types of multifunctionally activated synthetic polymers include difunctionally activated, tetrafunctionally activated, and star-branched polymers.
• Multifunctionally activated synthetic polymers for use in the present invention must contain at least two, more preferably, at least three, functional groups in order to form a three-dimensional crosslinked network with synthetic polymers containing multiple nucleophilic groups (i.e., "multi- nucleophilic polymers"). In other words, they must be at least difunctionally activated, and are more preferably trifunctionally or tetrafunctionally activated. If the first synthetic polymer is a difunctionally activated synthetic polymer, the second synthetic polymer must contain three or more functional groups in order to obtain a three-dimensional crosslinked network. Most preferably, both the first and the second synthetic polymer contain at least three functional groups.
• Multi-nucleophilic polymers Synthetic polymers containing multiple nucleophilic groups are also referred to generically herein as "multi-nucleophilic polymers.”
• multi-nucleophilic polymers must contain at least two, more preferably, at least three, nucleophilic groups. If a synthetic polymer containing only two nucleophilic groups is used, a synthetic polymer containing three or more electrophilic groups must be used in order to obtain a three- dimensional crosslinked network.
• Preferred multi-nucleophilic polymers for use in the compositions and methods of the present invention include synthetic polymers that contain, or have been modified to contain, multiple nucleophilic groups such as primary amino groups and thiol groups.
• Preferred multi-nucleophilic polymers include: (i) synthetic polypeptides that have been synthesized to contain two or more primary amino groups or thiol groups; and (ii) polyethylene glycols that have been modified to contain two or more primary amino groups or thiol groups.
• reaction of a thiol group with an electrophilic group tends to proceed more slowly than reaction of a primary amino group with an electrophilic group.
• the m ⁇ lti-nucleophilic polypeptide is a synthetic polypeptide that has been synthesized to incorporate amino acid residues containing primary amino groups (such as lysine) and/or amino acids containing thiol groups (such as cysteine).
• Poly(lysine)s have been prepared having anywhere from 6 to about 4,000 primary amino groups, corresponding to molecular weights of about 870 to about 580,000.
• Poly(lysine)s for use in the present invention preferably have a molecular weight within the range of about 1 ,000 to about 300,000; more preferably, within the range of about 5,000 to about 100,000; most preferably, within the range of about 8,000 to about 15,000.
• Poly(lysine)s of varying molecular weights are commercially available from Peninsula Laboratories, Inc. (Belmont, Calif.) and Aldrich Chemical (Milwaukee, WI).
• Polyethylene glycol can be chemically modified to contain multiple primary amino or thiol groups according to methods set forth, for example, in Chapter 22 of Poly(ethylene Glycol) Chemistry: Biotechnical and Biomedical Applications, J. Milton Harris, ed., Plenum Press, N.Y. (1992). Polyethylene glycols which have been modified to contain two or more primary amino groups are referred to herein as "multi-amino PEGs.” Polyethylene glycols which have been modified to contain two or more thiol groups are referred to herein as "multi-thiol PEGs.” As used herein, the term "polyethylene glycol(s)" includes modified and or derivatized polyethylene glycol(s).
• Multi-amino PEGs useful in the present invention include Huntsman's Jeffamine diamines ("D” series) and triamines ("T” series), which contain two and three primary amino groups per molecule, respectively.
• Polyamines such as ethylenediamine (H 2 N-CH 2 -CH 2 -NH 2 ), tetramethylenediamine (H 2 N-(CH 2 ) 4 -NH 2 ), pentamethylenediamine (cadaverine) (H 2 N-(CH 2 ) 5 -NH 2 ), hexamethylenediamine (H 2 N-(CH 2 ) 6 -NH 2 ), di(2- aminoethyl)amine (HN-(CH 2 -CH 2 -NH 2 ) 2 ), and tris(2-aminoethyl)amine (N-(CH 2 - CH 2 -NH 2 ) 3 ) may also be used as the synthetic polymer containing multiple nucleophilic groups.
• ethylenediamine H 2 N-CH 2 -CH 2 -NH 2
• tetramethylenediamine H 2 N-(CH 2 ) 4 -NH 2
• pentamethylenediamine cadaverine
• Multi-electrophilic polymers Synthetic polymers containing multiple electrophilic groups are also referred to herein as "multi-electrophilic polymers.”
• the multifunctionally activated synthetic polymers must contain at least two, more preferably, at least three, electrophilic groups in order to form a three-dimensional crosslinked network with multi-nucleophilic polymers.
• Preferred multi-electrophilic polymers for use in the compositions of the invention are polymers which contain two or more succinimidyl groups capable of forming covalent bonds with nucleophilic groups on other molecules. Succinimidyl groups are highly reactive with materials containing primary amino (NH 2 ) groups, such as multi-amino PEG, poly(lysine), or collagen.
• Succinimidyl groups are slightly less reactive with materials containing thiol (SH) groups, such as multi-thiol PEG or synthetic polypeptides containing multiple cysteine residues.
• thiol (SH) groups such as multi-thiol PEG or synthetic polypeptides containing multiple cysteine residues.
• the term "containing two or more succinimidyl groups” is meant to encompass polymers which are preferably commercially available containing two or more succinimidyl groups, as well as those that must be chemically derivatized to contain two or more succinimidyl groups.
• succinimidyl group is intended to encompass sulfosuccinimidyl groups and other such variations of the "generic" succinimidyl group.
• PEG refers to polymers having the repeating structure (OCH 2 -CH 2 ) n . Structures for some specific, tetrafunctionally activated forms of PEG are shown in FIGS. 4 to 13 of U.S. Patent 5,874,500, incorporated herein by reference.
• PEGS examples include PEG succinimidyl propionate (SE-PEG), PEG succinimidyl succinamide (SSA-PEG), and PEG succinimidyl carbonate (SC-PEG).
• SE-PEG PEG succinimidyl propionate
• SSA-PEG PEG succinimidyl succinamide
• SC-PEG PEG succinimidyl carbonate
• the crosslinked matrix is formed in situ by reacting pentaerythritol poly(ethylene glycol)ether tetra-sulfhydryl] (4-armed thiol PEG) and pentaerythritol poly(ethylene glycol)ether tetra-succinimidyl glutarate] (4-armed NHS PEG) as reactive reagents. Structures for these reactants are shown in U.S. Patent 5,874,500.
• Each of these materials has a core with a structure that may be seen by adding ethylene oxide-derived residues to each of the hydroxyl groups in pentaerythritol, and then derivatizing the terminal hydroxyl groups (derived from the ethylene oxide) to contain either thiol groups (so as to form 4-armed thiol PEG) or N-hydroxysuccinimydyl groups (so as to form 4-armed NHS PEG), optionally with a linker group present between the ethylene oxide derived backbone and the reactive functional group, where this product is commercially available as COSEAL from Angiotech Pharmaceuticals Inc.
• a group "D" may be present in one or both of these molecules, as discussed in more detail below.
• preferred activated polyethylene glycol derivatives for use in the invention contain succinimidyl groups as the reactive group.
• different activating groups can be attached at sites along the length of the PEG molecule.
• PEG can be derivatized to form functionally activated PEG propionaldehyde (A-PEG), or functionally activated PEG glycidyl ether (E-PEG), or functionally activated PEG-isocyanate (l-PEG), or functionally activated PEG-vinylsulfone (V-PEG).
• Hydrophobic polymers can also be used to prepare the compositions of the present invention.
• Hydrophobic polymers for use in the present invention preferably contain, or can be derivatized to contain, two or more electrophilic groups, such as succinimidyl groups, most preferably, two, three, or four electrophilic groups.
• hydrophobic polymer refers to polymers which contain a relatively small proportion of oxygen or nitrogen atoms.
• Hydrophobic polymers which already contain two or more succinimidyl groups include, without limitation, disuccinimidyl suberate (DSS), bis(sulfosuccinimidyl) suberate (BS3), dithiobis(succinimidylpropionate) (DSP), bis(2-succinimidooxycarbonyloxy) ethyl sulfone (BSOCOES), and 3,3'- dithiobis(sulfosuccinimidylpropionate (DTSPP), and their analogs and derivatives.
• DSS disuccinimidyl suberate
• BS3 bis(sulfosuccinimidyl) suberate
• DSP dithiobis(succinimidylpropionate)
• BSOCOES bis(2-succinimidooxycarbonyloxy) ethyl sulfone
• DTSPP 3,3'- dithiobis(sulfosuccinimi
• Preferred hydrophobic polymers for use in the invention generally have a carbon chain that is no longer than about 14 carbons.
• Polymers having carbon chains substantially longer than 14 carbons generally have very poor solubility in aqueous solutions and, as such, have very long reaction times when mixed with aqueous solutions of synthetic polymers containing multiple nucleophilic groups.
• Certain polymers, such as polyacids, can be derivatized to contain two or more functional groups, such as succinimidyl groups.
• Polyacids for use in the present invention include, without limitation, trimethylolpropane-based tricarboxylic acid, di(trimethylol propane)-based tetracarboxylic acid, heptanedioic acid, octanedioic acid (suberic acid), and hexadecanedioic acid (thapsic acid). Many of these polyacids are commercially available from DuPont Chemical Company (Wilmington, DE).
• polyacids can be chemically derivatized to contain two or more succinimidyl groups by reaction with an appropriate molar amount of N-hydroxysuccinimide (NHS) in the presence of N,N'-dicyclohexylcarbodiimide (DCC).
• NHS N-hydroxysuccinimide
• DCC N,N'-dicyclohexylcarbodiimide
• Polyalcohols such as trimethylolpropane and di(trimethylol propane) can be converted to carboxylic acid form using various methods, then further derivatized by reaction with NHS in the presence of DCC to produce trifunctionally and tetrafunctionally activated polymers, respectively, as described in U.S. application Ser. No. 08/403,358.
• Polyacids such as heptanedioic acid (HOOC-(CH 2 ) 5 -COOH), octanedioic acid (HOOC-(CH 2 ) 6 - COOH), and hexadecanedioic acid (HOOC-(CH 2 ) ⁇ 4 -COOH) are derivatized by the addition of succinimidyl groups to produce difunctionally activated polymers.
• Polyamines such as ethylenediamine, tetramethylenediamine, pentamethylenediamine (cadaverine), hexamethylenediamine, bis (2- aminoethyl)amine, and tris(2-aminoethyl)amine can be chemically derivatized to polyacids, which can then be derivatized to contain two or more succinimidyl groups by reacting with the appropriate molar amounts of N- hydroxysuccinimide in the presence of DCC, as described in U.S. application Ser. No. 08/403,358. Many of these polyamines are commercially available from DuPont Chemical Company.
• the first synthetic polymer will contain multiple nucleophilic groups (represented below as “X”) and it will react with the second synthetic polymer containing multiple electrophilic groups (represented below as “Y”), resulting in a covalently bound polymer network, as follows: Polymer-X m + Polymer-Y n ⁇ Polymer-Z-Polymer wherein m ⁇ 2, n ⁇ 2, and m + n ⁇ 5; where exemplary X groups include -NH 2 , -SH, -OH, -PH 2 , CO-NH-
• X groups may be the same or different in polymer-X m ;
• X and Y may be the same or different, i.e., a synthetic polymer may have two different electrophilic groups, or two different nucleophilic groups, such as with glutathione.
• the backbone of at least one of the synthetic polymers comprises alkylene oxide residues, e.g., residues from ethylene oxide, propylene oxide, and mixtures thereof.
• the term 'backbone' refers to a significant portion of the polymer.
• the synthetic polymer containing alkylene oxide residues may be described by the formula X-polymer-X or Y-polymer-Y, wherein X and Y are as defined above, and the term "polymer” represents - (CH 2 CH 2 0) n - or -(CH(CH 3 )CH 2 0) n - or -(CH 2 -CH 2 -0) n -(CH(CH 3 )CH 2 -0)n-. In these cases the synthetic polymer would be difunctional.
• the required functional group X or Y is commonly coupled to the polymer backbone by a linking group (represented below as "Q”), many of which are known or possible.
• Q groups include -0-(CH 2 ) n -; -S-(CH 2 ) n -; -NH-(CH 2 ) n -; -0 2 C-NH-(CH 2 ) n -; -0 2 C-(CH 2 ) n -; -0 2 C-(CR 1 H) n -; and -0-R 2 -CO-NH-, which provide synthetic polymers of the partial structures: polymer-0-(CH 2 ) n -(X or Y); polymer-S-(CH 2 )n-(X or Y); polymer-NH-(CH 2 ) n -(X or Y); polymer-0 2 C-NH- (CH 2 ) n -(X or Y); polymer-0 2 C-NH- (CH 2 ) n -(X or Y); polymer-0 2 C-NH- (CH 2 ) n -(X or Y); polymer-0 2 C-NH
• n 1-10
• R 1 H or alkyl (i.e., CH 3 , C 2 H 5 , etc.);
• R 2 CH 2 , or CO-NH-CH 2 CH 2 ;
• Q-, and Q 2 may be the same or different.
• Q 2 OCH 2 CH 2 (there is no Q-i in this case);
• D An additional group, represented below as "D" can be inserted between the polymer and the linking group, if present.
• D group One purpose of such a D group is to affect the degradation rate of the crosslinked polymer composition in vivo, for example, to increase the degradation rate, or to decrease the degradation rate. This may be useful in many instances, for example, when drug has been incorporated into the matrix, and it is desired to increase or decrease polymer degradation rate so as to influence a drug delivery profile in the desired direction.
• An illustration of a crosslinking reaction involving first and second synthetic polymers each having D and Q groups is shown below.
• Some useful biodegradable groups "D" include polymers formed from one or more ⁇ -hydroxy acids, e.g., lactic acid, glycolic acid, and the cyclization products thereof (e.g., lactide, glycolide), ⁇ -caprolactone, and amino acids.
• the polymers may be referred to as polylactide, polyglycolide, poly(co- lactide-glycolide); poly- ⁇ -caprolactone, polypeptide (also known as poly amino acid, for example, various di- or tri-peptides) and poly(anhydride)s.
• a first synthetic polymer containing multiple nucleophilic groups is mixed with a second synthetic polymer containing multiple electrophilic groups. Formation of a three-dimensional crosslinked network occurs as a result of the reaction between the nucleophilic groups on the first synthetic polymer and the electrophilic groups on the second synthetic polymer.
• concentrations of the first synthetic polymer and the second synthetic polymer used to prepare the compositions of the present invention will vary depending upon a number of factors, including the types and molecular weights of the particular synthetic polymers used and the desired end use application.
• multi-amino PEG as the first synthetic polymer, it is preferably used at a concentration in the range of about 0.5 to about 20 percent by weight of the final composition, while the second synthetic polymer is used at a concentration in the range of about 0.5 to about 20 percent by weight of the final composition.
• a final composition having a total weight of 1 gram (1000 milligrams) would contain between about 5 to about 200 milligrams of multi-amino PEG, and between about 5 to about 200 milligrams of the second synthetic polymer.
• Use of higher concentrations of both first and second synthetic polymers will result in the formation of a more tightly crosslinked network, producing a stiffer, more robust gel.
• compositions intended for use in tissue augmentation will generally employ concentrations of first and second synthetic polymer that fall toward the higher end of the preferred concentration range.
• Compositions intended for use as bioadhesives or in adhesion prevention do not need to be as firm and may therefore contain lower polymer concentrations.
• the second synthetic polymer is generally stored and used in sterile, dry form to prevent the loss of crosslinking ability due to hydrolysis which typically occurs upon exposure of such electrophilic groups to aqueous media. Processes for preparing synthetic hydrophilic polymers containing multiple electrophylic groups in sterile, dry form are set forth in U.S. Patent 5,643,464.
• the dry synthetic polymer may be compression molded into a thin sheet or membrane, which can then be sterilized using gamma or, preferably, e-beam irradiation.
• the resulting dry membrane or sheet can be cut to the desired size or chopped into smaller size particulates.
• polymers containing multiple nucleophilic groups are generally not water-reactive and can therefore be stored in aqueous solution.
• one or both of the electrophilic- or nucleophilic-terminated polymers described above can be combined with a synthetic or naturally occurring polymer. The presence of the synthetic or naturally occurring polymer may enhance the mechanical and/or adhesive properties of the in situ forming compositions.
• Naturally occurring polymers, and polymers derived from naturally occurring polymer that may be included in in situ forming materials include naturally occurring proteins, such as collagen, collagen derivatives (such as methylated collagen), fibrinogen, thrombin, albumin, fibrin, and derivatives of and naturally occurring polysaccharides, such as glycosaminoglycans, including deacetylated and desulfated glycosaminoglycan derivatives.
• a composition comprising naturally-occurring protein and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising collagen and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising methylated collagen and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising fibrinogen and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising thrombin and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising albumin and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising fibrin and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising naturally occurring polysaccharide and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising glycosaminoglycan and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising deacetylated glycosaminoglycan and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising desulfated glycosaminoglycan and both of the first and second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising naturally-occurring protein and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising collagen and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising methylated collagen and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising fibrinogen and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising thrombin and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising albumin and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising fibrin and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising naturally occurring polysaccharide and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising glycosaminoglycan and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising deacetylated glycosaminoglycan and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising desulfated glycosaminoglycan and the first synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising naturally-occurring protein and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising collagen and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising methylated collagen and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising fibrinogen and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising thrombin and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising albumin and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising fibrin and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising naturally occurring polysaccharide and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising glycosaminoglycan and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising deacetylated glycosaminoglycan and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• a composition comprising desulfated glycosaminoglycan and the second synthetic polymer as described above is used to form the crosslinked matrix according to the present invention.
• protein or polysaccharide components which contain functional groups that can react with the functional groups on multiple activated synthetic polymers can result in formation of a crosslinked synthetic polymer-naturally occurring polymer matrix upon mixing and/or crosslinking of the synthetic polymer(s).
• the naturally occurring polymer protein or polysaccharide
• the electrophilic groups on the second synthetic polymer will react with the primary amino groups on these components, as well as the nucleophilic groups on the first synthetic polymer, to cause these other components to become part of the polymer matrix.
• lysine-rich proteins such as collagen may be especially reactive with electrophilic groups on synthetic polymers.
• the naturally occurring protein is polymer may be collagen.
• collagen refers to all forms of collagen, including those which have been processed or otherwise modified and is intended to encompass collagen of any type, from any source, including, but not limited to, collagen extracted from tissue or produced recombinantly, collagen analogues, collagen derivatives, modified collagens, and denatured collagens, such as gelatin.
• collagen from any source may be included in the compositions of the invention; for example, collagen may be extracted and purified from human or other mammalian source, such as bovine or porcine corium and human placenta, or may be recombinantly or otherwise produced.
• the preparation of purified, substantially non-antigenic collagen in solution from bovine skin is well known in the art. U.S.
• Patent No. 5,428,022 discloses methods of extracting and purifying collagen from the human placenta.
• U.S. Patent No. 5,667,839 discloses methods of producing recombinant human collagen in the milk of transgenic animals, including transgenic cows.
• Collagen of any type including, but not limited to, types I, II, III, IV, or any combination thereof, may be used in the compositions of the invention, although type I is generally preferred.
• Either atelopeptide or telopeptide-containing collagen may be used; however, when collagen from a xenogeneic source, such as bovine collagen, is used, atelopeptide collagen is generally preferred, because of its reduced immunogenicity compared to telopeptide-containing collagen.
• Collagen that has not been previously crosslinked by methods such as heat, irradiation, or chemical crosslinking agents is preferred for use in the compositions of the invention, although previously crosslinked collagen may be used.
• Non-crosslinked atelopeptide fibrillar collagen is commercially available from Inamed Aesthetics (Santa Barbara, CA) at collagen concentrations of 35 mg/ml and 65 mg/ml under the trademarks ZYDERM I Collagen and ZYDERM II Collagen, respectively.
• Glutaraldehyde crosslinked atelopeptide fibrillar collagen is commercially available from Inamed Corporation (Santa Barbara, CA) at a collagen concentration of 35 mg/ml under the trademark ZYPLAST Collagen.
• Collagens for use in the present invention are generally in aqueous suspension at a concentration between about 20 mg/ml to about 120 mg/ml; preferably, between about 30 mg/ml to about 90 mg/ml. Because of its tacky consistency, nonfibrillar collagen may be preferred for use in compositions that are intended for use as bioadhesives.
• nonfibrillar collagen refers to any modified or unmodified collagen material that is in substantially nonfibrillar form at pH 7, as indicated by optical clarity of an aqueous suspension of the collagen. Collagen that is already in nonfibrillar form may be used in the compositions of the invention.
• nonfibrillar collagen is intended to encompass collagen types that are nonfibrillar in native form, as well as collagens that have been chemically modified such that they are in nonfibrillar form at or around neutral pH.
• Collagen types that are nonfibrillar (or microfibrillar) in native form include types IV, VI, and VII.
• Chemically modified collagens that are in nonfibrillar form at neutral pH include succinylated collagen and methylated collagen, both of which can be prepared according to the methods described in U.S. Pat. No. 4,164,559, issued Aug. 14, 1979, to Miyata et al., which is hereby incorporated by reference in its entirety.
• methylated collagen is particularly preferred for use in bioadhesive compositions, as disclosed in U.S. application Ser. No. 08/476,825.
• Collagens for use in the crosslinked polymer compositions of the present invention may start out in fibrillar form, then be rendered nonfibrillar by the addition of one or more fiber disassembly agent.
• the fiber disassembly agent must be present in an amount sufficient to render the collagen substantially nonfibrillar at pH 7, as described above.
• Fiber disassembly agents for use in the present invention include, without limitation, various biocompatible alcohols, amino acids (e.g., arginine), inorganic salts (e.g., sodium chloride and potassium chloride), and carbohydrates (e.g., various sugars including sucrose).
• the polymer may be collagen or a collagen derivative, for example methylated collagen.
• an in situ forming composition uses pentaerythritol poly(ethylene glycol)ether tetra-sulfhydryl] (4- armed thiol PEG), pentaerythritol poly( ethylene glycol)ether tetra-succinimidyl glutarate] (4-armed NHS PEG) and methylated collagen as the reactive reagents.
• This composition when mixed with the appropriate buffers can produce a crosslinked hydrogel.
• the naturally occurring polymer may be a glycosaminoglycan.
• Glycosaminoglycans e.g., hyaluronic acid
• the glycosaminoglycan may be derivatized.
• glycosaminoglycans can be chemically derivatized by, e.g., deacetylation, desulfation, or both in order to contain primary amino groups available for reaction with electrophilic groups on synthetic polymer molecules.
• Glycosaminoglycans that can be derivatized according to either or both of the aforementioned methods include the following: hyaluronic acid, chondroitin sulfate A, chondroitin sulfate B (dermatan sulfate), chondroitin sulfate C, chitin (can be derivatized to chitosan), keratan sulfate, keratosulfate, and heparin.
• Derivatization of glycosaminoglycans by deacetylation and/or desulfation and covalent binding of the resulting glycosaminoglycan derivatives with synthetic hydrophilic polymers is described in further detail in commonly assigned, allowed U.S.
• the collagen is added to the first synthetic polymer, then the collagen and first synthetic polymer are mixed thoroughly to achieve a homogeneous composition.
• the second synthetic polymer is then added and mixed into the collagen/first synthetic polymer mixture, where it will covalently bind to primary amino groups or thiol groups on the first synthetic polymer and primary amino groups on the collagen, resulting in the formation of a homogeneous crosslinked network.
• Various deacetylated and/or desulfated glycosaminoglycan derivatives can be incorporated into the composition in a similar manner as that described above for collagen.
• the introduction of hydrocolloids such as carboxymethylcellulose may promote tissue adhesion and/or swellability.
• compositions of the present invention having two synthetic polymers may be administered before, during or after crosslinking of the first and second synthetic polymer.
• the point at which crosslinking has reached equilibrium is defined herein as the point at which the composition no longer feels tacky or sticky to the touch.
• the first synthetic polymer and second synthetic polymer may be contained within separate barrels of a dual-compartment syringe.
• the two synthetic polymers do not actually mix until the point at which the two polymers are extruded from the tip of the syringe needle into the patient's tissue.
• This allows the vast majority of the crosslinking reaction to occur in situ, avoiding the problem of needle blockage which commonly occurs if the two synthetic polymers are mixed too early and crosslinking between the two components is already too advanced prior to delivery from the syringe needle.
• the use of a dual-compartment syringe, as described above, allows for the use of smaller diameter needles, which is advantageous when performing soft tissue augmentation in delicate facial tissue, such as that surrounding the eyes.
• first synthetic polymer and second synthetic polymer may be mixed according to the methods described above prior to delivery to the tissue site, then injected to the desired tissue site immediately (preferably, within about 60 seconds) following mixing.
• first synthetic polymer and second synthetic polymer are mixed, then extruded and allowed to crosslink into a sheet or other solid form. The crosslinked solid is then dehydrated to remove substantially all unbound water.
• the resulting dried solid may be ground or comminuted into particulates, then suspended in a nonaqueous fluid carrier, including, without limitation, hyaluronic acid, dextran sulfate, dextran, succinylated noncrosslinked collagen, methylated noncrosslinked collagen, glycogen, glycerol, dextrose, maltose, triglycerides of fatty acids (such as corn oil, soybean oil, and sesame oil), and egg yolk phospholipid.
• a nonaqueous fluid carrier including, without limitation, hyaluronic acid, dextran sulfate, dextran, succinylated noncrosslinked collagen, methylated noncrosslinked collagen, glycogen, glycerol, dextrose, maltose, triglycerides of fatty acids (such as corn oil, soybean oil, and sesame oil), and egg yolk phospholipid.
• the suspension of particulates can be injected through a small- gauge needle to a tissue site
• the first and/or second synthetic polymers may be combined with a hydrophilic polymer, e.g., collagen or methylated collagen, to form a composition useful in the present invention.
• a hydrophilic polymer e.g., collagen or methylated collagen
• the compositions useful in the present invention include a hydrophilic polymer in combination with two or more crosslinkable components. This embodiment is described in further detail in this section.
• the hydrophilic Polymer component may be a synthetic or naturally occurring hydrophilic polymer.
• Naturally occurring hydrophilic polymers include, but are not limited to: proteins such as collagen and derivatives thereof, fibronectin, albumins, globulins, fibrinogen, and fibrin, with collagen particularly preferred; carboxylated polysaccharides such as polymannuronic acid and polygalacturonic acid; aminated polysaccharides, particularly the glycosaminoglycans, e.g., hyaluronic acid, chitin, chondroitin sulfate A, B, or C, keratin sulfate, keratosulfate and heparin; and activated polysaccharides such as dextran and starch derivatives.
• Collagen e.g., methylated collagen
• glycosaminoglycans are preferred naturally occurring hydrophilic polymers for use herein.
• collagen from any source may be used in the composition of the method; for example, collagen may be extracted and purified from human or other mammalian source, such as bovine or porcine corium and human placenta, or may be recombinantly or otherwise produced.
• the preparation of purified, substantially non-antigenic collagen in solution from bovine skin is well known in the art. See, e.g., U.S. Pat. No. 5,428,022, to Palefsky et al., which discloses methods of extracting and purifying collagen from the human placenta. See also U.S. Patent No.
• collagen or "collagen material” as used herein refers to all forms of collagen, including those that have been processed or otherwise modified.
• Collagen of any type including, but not limited to, types I, II, III, IV, or any combination thereof, may be used in the compositions of the invention, although type I is generally preferred.
• Either atelopeptide or telopeptide- containing collagen may be used; however, when collagen from a source, such as bovine collagen, is used, atelopeptide collagen is generally preferred, because of its reduced immunogenicity compared to telopeptide-containing collagen.
• Collagen that has not been previously crosslinked by methods such as heat, irradiation, or chemical crosslinking agents is preferred for use in the compositions of the invention, although previously crosslinked collagen may be used.
• Non-crosslinked atelopeptide fibrillar collagen is commercially available from McGhan Medical Corporation (Santa Barbara, Calif.) at collagen concentrations of 35 mg/ml and 65 mg/ml under the trademarks ZYDERM ® I Collagen and ZYDERM ® II Collagen, respectively.
• Glutaraldehyde-crosslinked atelopeptide fibrillar collagen is commercially available from McGhan Medical
• Collagens for use in the present invention are generally, although not necessarily, in aqueous suspension at a concentration between about 20 mg/ml to about 120 mg/ml, preferably between about 30 mg/ml to about 90 mg/ml.
• intact collagen is preferred, denatured collagen, commonly known as gelatin, can also be used in the compositions of the invention.
• Gelatin may have the added benefit of being degradable faster than collagen. Because of its greater surface area and greater concentration of reactive groups, nonfibrillar collagen is generally preferred. The term
• nonfibrillar collagen refers to any modified or unmodified collagen material that is in substantially nonfibrillar form at pH 7, as indicated by optical clarity of an aqueous suspension of the collagen. Collagen that is already in nonfibrillar form may be used in the compositions of the invention.
• nonfibrillar collagen is intended to encompass collagen types that are nonfibrillar in native form, as well as collagens that have been chemically modified such that they are in nonfibrillar form at or around neutral pH.
• Collagen types that are nonfibrillar (or microfibrillar) in native form include types IV, VI, and VII.
• Chemically modified collagens that are in nonfibrillar form at neutral pH include succinylated collagen, propylated collagen, ethylated collagen, methylated collagen, and the like, both of which can be prepared according to the methods described in U.S. Pat. No. 4,164,559, to Miyata et al., which is hereby incorporated by reference in its entirety. Due to its inherent tackiness, methylated collagen is particularly preferred, as disclosed in U.S. Patent No. 5,614,587 to Rhee et al. Collagens for use in the crosslinkable compositions of the present invention may start out in fibrillar form, then be rendered nonfibrillar by the addition of one or more fiber disassembly agents.
• Fiber disassembly agent must be present in an amount sufficient to render the collagen substantially nonfibrillar at pH 7, as described above.
• Fiber disassembly agents for use in the present invention include, without limitation, various biocompatible alcohols, amino acids, inorganic salts, and carbohydrates, with biocompatible alcohols being particularly preferred.
• Preferred biocompatible alcohols include glycerol and propylene glycol.
• Non-biocompatible alcohols, such as ethanol, methanol, and isopropanol are not preferred for use in the present invention, due to their potentially deleterious effects on the body of the patient receiving them.
• Preferred amino acids include arginine.
• Preferred inorganic salts include sodium chloride and potassium chloride.
• carbohydrates such as various sugars including sucrose
• they are not as preferred as other types of fiber disassembly agents because they can have cytotoxic effects in vivo.
• fibrillar collagen has less surface area and a lower concentration of reactive groups than nonfibrillar, fibrillar collagen is less preferred.
• fibrillar collagen, or mixtures of nonfibrillar and fibrillar collagen may be preferred for use in compositions intended for long-term persistence in vivo, if optical clarity is not a requirement.
• Synthetic hydrophilic polymers may also be used in the present invention.
• Useful synthetic hydrophilic polymers include, but are not limited to: polyalkylene oxides, particularly polyethylene glycol and poly(ethylene oxide)- poly(propylene oxide) copolymers, including block and random copolymers; polyols such as glycerol, polyglycerol (particularly highly branched polyglycerol), propylene glycol and trimethylene glycol substituted with one or more polyalkylene oxides, e.g., mono-, di- and tri-polyoxyethylated glycerol, mono- and di-polyoxyethylated propylene glycol, and mono- and di- polyoxyethylated trimethylene glycol; polyoxyethylated sorbitol, polyoxyethylated glucose; acrylic acid polymers and analogs and copolymers thereof, such as polyacrylic acid per se, polymethacrylic acid, poly(hydroxyethyl-methacrylate), poly(hydroxyethylacrylate), poly(methylaIky
• the compositions of the invention also comprise a plurality of crosslinkable components.
• Each of the crosslinkable components participates in a reaction that results in a crosslinked matrix.
• the crosslinkable components Prior to completion of the crosslinking reaction, the crosslinkable components provide the necessary adhesive qualities that enable the methods of the invention.
• the crosslinkable components are selected so that crosslinking gives rise to a biocompatible, nonimmunogenic matrix useful in a variety of contexts including adhesion prevention, biologically active agent delivery, tissue augmentation, and other applications.
• the crosslinkable components of the invention comprise: a component A, which has m nucleophilic groups, wherein m > 2 and a component B, which has n electrophilic groups capable of reaction with the m nucleophilic groups, wherein n > 2 and m + n > 4.
• An optional third component, optional component C, which has at least one functional group that is either electrophilic and capable of reaction with the nucleophilic groups of component A, or nucleophilic and capable of reaction with the electrophilic groups of component B may also be present.
• the total number of functional groups present on components A, B and C, when present, in combination is > 5; that is, the total functional groups given by m + n + p must be > 5, where p is the number of functional groups on component C and, as indicated, is > 1.
• Each of the components is biocompatible and nonimmunogenic, and at least one component is comprised of a hydrophilic polymer.
• the composition may contain additional crosslinkable components D, E, F, etc., having one or more reactive nucleophilic or electrophilic groups and thereby participate in formation of the crosslinked biomaterial via covalent bonding to other components.
• the m nucleophilic groups on component A may all be the same, or, alternatively, A may contain two or more different nucleophilic groups.
• the n electrophilic groups on component B may all be the same, or two or more different electrophilic groups may be present.
• the functional group(s) on optional component C if nucleophilic, may or may not be the same as the nucleophilic groups on component A, and, conversely, if electrophilic, the functional group(s) on optional component C may or may not be the same as the electrophilic groups on component B.
• the components may be represented by the structural formulae (I) R 1 (-[Q 1 ] q -X) m (component A), (II) R 2 (-[Q 2 ] r Y)n (component B), and (III) R 3 (-[Q 3 ] S -Fn)p (optional component C), wherein: R 1 , R 2 and R 3 are independently selected from the group consisting of C 2 to C- ⁇ hydrocarbyl, heteroatom-containing C 2 to C 14 hydrocarbyl, hydrophilic polymers, and hydrophobic polymers, providing that at least one of R 1 , R 2 and R 3 is a hydrophilic polymer, preferably a synthetic hydrophilic polymer; X represents one of the m nucleophilic groups of component A, and the various X moieties on A may be the same or different; Y represents one of the n electrophilic groups of component B, and the various Y moieties on A may be the same or different; Fn represents a functional
• Reactive Groups may be virtually any nucleophilic group, so long as reaction can occur with the electrophilic group Y.
• Y may be virtually any electrophilic group, so long as reaction can take place with X.
• the only limitation is a practical one, in that reaction between X and Y should be fairly rapid and take place automatically upon admixture with an aqueous medium, without need for heat or potentially toxic or non-biodegradable reaction catalysts or other chemical reagents. It is also preferred although not essential that reaction occur without need for ultraviolet or other radiation.
• the reactions between X and Y should be complete in under 60 minutes, preferably under 30 minutes. Most preferably, the reaction occurs in about 5 to 15 minutes or less.
• nucleophilic groups suitable as X include, but are not limited to, -NH 2 , -NHR 4 , -N(R 4 ) 2 , -SH, -OH, -COOH, -C 6 H 4 -OH, -PH 2 , -PHR 5 , - P(R 5 ) 2 , -NH-NH 2 , -CO-NH-NH 2 , -C 5 H 4 N, etc.
• R 4 and R 5 are hydrocarbyl, typically alkyl or monocyclic aryl, preferably alkyl, and most preferably lower alkyl.
• Organometallic moieties are also useful nucleophilic groups for the purposes of the invention, particularly those that act as carbanion donors.
• Organometallic nucleophiles are not, however, preferred.
• organometallic moieties include: Grignard functionalities -R 6 MgHal wherein R 6 is a carbon atom (substituted or unsubstituted), and Hal is halo, typically bromo, iodo or chloro, preferably bromo; and lithium-containing functionalities, typically alkyllithium groups; sodium-containing functionalities. It will be appreciated by those of ordinary skill in the art that certain nucleophilic groups must be activated with a base so as to be capable of reaction with an electrophile.
• the composition when there are nucleophilic sulfhydryl and hydroxyl groups in the crosslinkable composition, the composition must be admixed with an aqueous base in order to remove a proton and provide an -S " or -O " species to enable reaction with an electrophile.
• a nonnucleophilic base is preferred.
• the base may be present as a component of a buffer solution. Suitable bases and corresponding crosslinking reactions are described infra in Section E.
• the selection of electrophilic groups provided within the crosslinkable composition, i.e., on component B, must be made so that reaction is possible with the specific nucleophilic groups.
• the Y groups are selected so as to react with amino groups.
• the X moieties are sulfhydryl moieties
• the corresponding electrophilic groups are sulfhydryl-reactive groups, and the like.
• a carboxylic acid group per se is not susceptible to reaction with a nucleophilic amine
• components containing carboxylic acid groups must be activated so as to be amine-reactive. Activation may be accomplished in a variety of ways, but often involves reaction with a suitable hydroxyl-containing compound in the presence of a dehydrating agent such as dicyclohexylcarbodiimide (DCC) or dicyclohexylurea (DHU).
• a dehydrating agent such as dicyclohexylcarbodiimide (DCC) or dicyclohexylurea (DHU).
• a carboxylic acid can be reacted with an alkoxy-substituted N- hydroxy-succinimide or N-hydroxysulfosuccinimide in the presence of DCC to form reactive electrophilic groups, the N-hydroxysuccinimide ester and the N- hydroxysulfosuccinimide ester, respectively.
• Carboxylic acids may also be activated by reaction with an acyl halide such as an acyl chloride (e.g., acetyl chloride), to provide a reactive anhydride group.
• a carboxylic acid may be converted to an acid chloride group using, e.g., thionyl chloride or an acyl chloride capable of an exchange reaction.
• thionyl chloride or an acyl chloride capable of an exchange reaction Specific reagents and procedures used to carry out such activation reactions will be known to those of ordinary skill in the art and are described in the pertinent texts and literature.
• the electrophilic groups present on Y are groups that react with a sulfhydryl moiety.
• Such reactive groups include those that form thioester linkages upon reaction with a sulfhydryl group, such as those described in PCT Publication No. WO 00/62827 to Wallace et al.
• such "sulfhydryl reactive" groups include, but are not limited to: mixed anhydrides; ester derivatives of phosphorus; ester derivatives of p-nitrophenol, p-nitrothiophenol and pentafluorophenol; esters of substituted hydroxylamines, including N-hydroxyphthalimide esters, N- hydroxysuccinimide esters, N-hydroxysulfosuccinimide esters, and N- hydroxyglutarimide esters; esters of 1-hydroxybenzotriazole; 3-hydroxy-3,4- dihydro-benzotriazin-4-one; 3-hydroxy-3,4-dihydro-quinazoline-4-one; carbonylimidazole derivatives; acid chlorides; ketenes; and isocyanates.
• auxiliary reagents can also be used to facilitate bond formation, e.g., 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide can be used to facilitate coupling of sulfhydryl groups to carboxyl-containing groups.
• sulfhydryl reactive groups that form thioester linkages various other sulfhydryl reactive functionalities can be utilized that form other types of linkages. For example, compounds that contain methyl imidate derivatives form imido-thioester linkages with sulfhydryl groups.
• sulfhydryl reactive groups can be employed that form disulfide bonds with sulfhydryl groups; such groups generally have the structure -S-S-Ar where Ar is a substituted or unsubstituted nitrogen-containing heteroaromatic moiety or a non-heterocyclic aromatic group substituted with an electron- withdrawing moiety, such that Ar may be, for example, 4-pyridinyl, o- nitrophenyl, m-nitrophenyl, p-nitrophenyl, 2,4-dinitrophenyl, 2-nitro-4-benzoic acid, 2-nitro-4-py rid inyl , etc.
• auxiliary reagents i.e., mild oxidizing agents such as hydrogen peroxide
• sulfhydryl reactive groups forms thioether bonds with sulfhydryl groups.
• groups include, inter alia, maleimido, substituted maleimido, haloalkyl, epoxy, imino, and aziridino, as well as olefins (including conjugated olefins) such as ethenesulfonyl, etheneimino, acrylate, methacrylate, and ⁇ , ⁇ -unsaturated aldehydes and ketones.
• This class of sulfhydryl reactive groups are particularly preferred as the thioether bonds may provide faster crosslinking and longer in vivo stability.
• the electrophilic functional groups on the remaining component(s) must react with hydroxyl groups.
• the hydroxyl group may be activated as described above with respect to carboxylic acid groups, or it may react directly in the presence of base with a sufficiently reactive electrophile such as an epoxide group, an aziridine group, an acyl halide, or an anhydride.
• suitable electrophilic functional groups for reaction therewith are those containing carbonyl groups, including, by way of example, ketones and aldehydes. It will also be appreciated that certain functional groups can react as nucleophiles or as electrophiles, depending on the selected reaction partner and/or the reaction conditions.
• a carboxylic acid group can act as a nucleophile in the presence of a fairly strong base, but generally acts as an electrophile allowing nucleophilic attack at the carbonyl carbon and concomitant replacement of the hydroxyl group with the incoming nucleophile.
• covalent linkages in the crosslinked structure that result upon covalent binding of specific nucleophilic components to specific electrophilic components in the crosslinkable composition include, solely by way of example, the following (the optional linking groups Q 1 and Q 2 are omitted for clarity):
• Linking Groups The functional groups X and Y and FN on optional component C may be directly attached to the compound core (R 1 , R 2 or R 3 on optional component C, respectively), or they may be indirectly attached through a linking group, with longer linking groups also termed "chain extenders.”
• chain extenders In structural formulae (I), (II) and (III), the optional linking groups are represented by Q 1 , Q 2 and Q 3 , wherein the linking groups are present when q, r and s are equal to 1 (with R, X, Y, Fn, m n and p as defined previously). Suitable linking groups are well known in the art. See, for example, International Patent Publication No. WO 97/22371.
• Linking groups are useful to avoid steric hindrance problems that are sometimes associated with the formation of direct linkages between molecules.
• Linking groups may additionally be used to link several multifunctionally activated compounds together to make larger molecules.
• a linking group can be used to alter the degradative properties of the compositions after administration and resultant gel formation.
• linking groups can be incorporated into components A, B, or optional component C to promote hydrolysis, to discourage hydrolysis, or to provide a site for enzymatic degradation.
• linking groups that provide hydrolyzable sites, include, inter alia: ester linkages; anhydride linkages, such as obtained by incorporation of glutarate and succinate; ortho ester linkages; ortho carbonate linkages such as trimethylene carbonate; amide linkages; phosphoester linkages; ⁇ -hydroxy acid linkages, such as may be obtained by incorporation of lactic acid and glycolic acid; lactone-based linkages, such as may be obtained by incorporation of caprolactone, valerolactone, ⁇ -butyrolactone and p- dioxanone; and amide linkages such as in a dimeric, oligomeric, or poly(amino acid) segment.
• non-degradable linking groups include succinimide, propionic acid and carboxymethylate linkages. See, for example, PCT WO 99/07417.
• enzymatically degradable linkages include Leu-Gly-Pro-Ala, which is degraded by collagenase; and Gly-Pro-Lys, which is degraded by plasmin.
• Linking groups can also enhance or suppress the reactivity of the various nucleophilic and electrophilic groups. For example, electron- withdrawing groups within one or two carbons of a sulfhydryl group would be expected to diminish its effectiveness in coupling, due to a lowering of nucleophilicity. Carbon-carbon double bonds and carbonyl groups will also have such an effect.
• n is generally in the range of 1 to about 10
• R 7 is generally hydrocarbyl, typically alkyl or aryl, preferably alkyl, and most preferably lower alkyl
• R 8 is hydrocarbylene, heteroatom-containing hydrocarbylene, substituted hydrocarbylene, or substituted heteroatom- containing hydrocarbylene) typically alkylene or arylene (again, optionally substituted and/or containing a heteroatom), preferably lower alkylene (e.g., methylene, ethylene, n-propylene, n-butylene, etc.), phenylene, or amidoalkylene (e.g., -(CO)-NH-CH 2 ).
• lower alkylene e.g., methylene, ethylene, n-propylene, n-butylene, etc.
• phenylene or amidoalkylene (e.g., -(CO)-NH-CH 2 ).
• linking groups are as follows: If higher molecular weight components are to be used, they preferably have biodegradable linkages as described above, so that fragments larger than 20,000 mol. wt. are not generated during resorption in the body. In addition, to promote water miscibility and/or solubility, it may be desired to add sufficient electric charge or hydrophilicity. Hydrophilic groups can be easily introduced using known chemical synthesis, so long as they do not give rise to unwanted swelling or an undesirable decrease in compressive strength. In particular, polyalkoxy segments may weaken gel strength.
• the Component Core The "core" of each crosslinkable component is comprised of the molecular structure to which the nucleophilic or electrophilic groups are bound.
• the "core” groups are R 1 , R 2 and R 3 .
• Each molecular core of the reactive components of the crosslinkable composition is generally selected from synthetic and naturally occurring hydrophilic polymers, hydrophobic polymers, and C 2 -C ⁇ 4 hydrocarbyl groups zero to 2 heteroatoms selected from N, O and S, with the proviso that at least one of the crosslinkable components A, B, and optionally C, comprises a molecular core of a synthetic hydrophilic polymer.
• at least one of A and B comprises a molecular core of a synthetic hydrophilic polymer.
• the crosslinkable component(s) is (are) hydrophilic polymers.
• hydrophilic polymer refers to a synthetic polymer having an average molecular weight and composition effective to render the polymer "hydrophilic" as defined above.
• synthetic crosslinkable hydrophilic polymers useful herein include, but are not limited to: polyalkylene oxides, particularly polyethylene glycol and poly( ethylene oxide)-poly(propylene oxide) copolymers, including block and random copolymers; polyols such as glycerol, polyglycerol (particularly highly branched polyglycerol), propylene glycol and trimethylene glycol substituted with one or more polyalkylene oxides, e.g., mono-, di- and tri-polyoxyethylated glycerol, mono- and di-polyoxyethylated propylene glycol, and mono- and di- polyoxyethylated trimethylene glycol; polyoxyethylated sorbitol, polyoxyethylated glucose; acrylic acid polymers and analogs and copolymers thereof, such as polyacrylic acid per se, polymethacrylic acid, poly(hydroxyethyl-methacrylate), poly(hydroxyethyl)
• the synthetic crosslinkable hydrophilic polymer may be a homopolymer, a block copolymer, a random copolymer, or a graft copolymer.
• the polymer may be linear or branched, and if branched, may be minimally to highly branched, dendrimeric, hyperbranched, or a star polymer.
• the polymer may include biodegradable segments and blocks, either distributed throughout the polymer's molecular structure or present as a single block, as in a block copolymer.
• Biodegradable segments are those that degrade so as to break covalent bonds. Typically, biodegradable segments are segments that are hydrolyzed in the presence of water and/or enzymatically cleaved in situ. Biodegradable segments may be composed of small molecular segments such as ester linkages, anhydride linkages, ortho ester linkages, ortho carbonate linkages, amide linkages, phosphonate linkages, etc. Larger biodegradable "blocks" will generally be composed of oligomeric or polymeric segments incorporated within the hydrophilic polymer.
• Illustrative oligomeric and polymeric segments that are biodegradable include, by way of example, poly(amino acid) segments, poly(orthoester) segments, poly(orthocarbonate) segments, and the like.
• Other suitable synthetic crosslinkable hydrophilic polymers include chemically synthesized polypeptides, particularly polynucleophilic polypeptides that have been synthesized to incorporate amino acids containing primary amino groups (such as lysine) and/or amino acids containing thiol groups (such as cysteine).
• Poly(lysine) a synthetically produced polymer of the amino acid lysine (145 MW), is particularly preferred.
• Poly(lysine)s have been prepared having anywhere from 6 to about 4,000 primary amino groups, corresponding to molecular weights of about 870 to about 580,000.
• Poly(lysine)s for use in the present invention preferably have a molecular weight within the range of about 1 ,000 to about 300,000, more preferably within the range of about 5,000 to about 100,000, and most preferably, within the range of about 8,000 to about 15,000.
• Poly(lysine)s of varying molecular weights are commercially available from Peninsula Laboratories, Inc. (Belmont, Calif.).
• the synthetic crosslinkable hydrophilic polymer may be a homopolymer, a block copolymer, a random copolymer, or a graft copolymer.
• the polymer may be linear or branched, and if branched, may be minimally to highly branched, dendrimeric, hyperbranched, or a star polymer.
• the polymer may include biodegradable segments and blocks, either distributed throughout the polymer's molecular structure or present as a single block, as in a block copolymer.
• Biodegradable segments are those that degrade so as to break covalent bonds.
• biodegradable segments are segments that are hydrolyzed in the presence of water and/or enzymatically cleaved in situ.
• Biodegradable segments may be composed of small molecular segments such as ester linkages, anhydride linkages, ortho ester linkages, ortho carbonate linkages, amide linkages, phosphonate linkages, etc.
• Larger biodegradable "blocks" will generally be composed of oligomeric or polymeric segments incorporated within the hydrophilic polymer.
• Illustrative oligomeric and polymeric segments that are biodegradable include, by way of example, poly(amino acid) segments, poly(orthoester) segments, poly(orthocarbonate) segments, and the like.
• preferred synthetic crosslinkable hydrophilic polymers are polyethylene glycol (PEG) and polyglycerol (PG), particularly highly branched polyglycerol.
• PEG polyethylene glycol
• PG polyglycerol
• Various forms of PEG are extensively used in the modification of biologically active molecules because PEG lacks toxicity, antigenicity, and immunogenicity (i.e., is biocompatible), can be formulated so as to have a wide range of solubilities, and do not typically interfere with the enzymatic activities and/or conformations of peptides.
• a particularly preferred synthetic crosslinkable hydrophilic polymer for certain applications is a polyethylene glycol (PEG) having a molecular weight within the range of about 100 to about 100,000 mol. wt., although for highly branched PEG, far higher molecular weight polymers can be employed - up to 1 ,000,000 or more - providing that biodegradable sites are incorporated ensuring that all degradation products will have a molecular weight of less than about 30,000.
• the preferred molecular weight is about 1 ,000 to about 20,000 mol. wt., more preferably within the range of about 7,500 to about 20,000 mol. wt.
• the polyethylene glycol has a molecular weight of approximately 10,000 mol. wt.
• Naturally occurring crosslinkable hydrophilic polymers include, but are not limited to: proteins such as collagen, fibronectin, albumins, globulins, fibrinogen, and fibrin, with collagen particularly preferred; carboxylated polysaccharides such as polymannuronic acid and polygalacturonic acid; aminated polysaccharides, particularly the glycosaminoglycans, e.g., hyaluronic acid, chitin, chondroitin sulfate A, B, or C, keratin sulfate, keratosulfate and heparin; and activated polysaccharides such as dextran and starch derivatives.
• proteins such as collagen, fibronectin, albumins, globulins, fibrinogen, and fibrin, with collagen particularly preferred
• carboxylated polysaccharides such as polymannuronic acid and polygalacturonic acid
• aminated polysaccharides particularly the glycosaminoglycans
• Collagen and glycosaminoglycans are examples of naturally occurring hydrophilic polymers for use herein, with methylated collagen being a preferred hydrophilic polymer.
• Any of the hydrophilic polymers herein must contain, or be activated to contain, functional groups, i.e., nucleophilic or electrophilic groups, which enable crosslinking. Activation of PEG is discussed below; it is to be understood, however, that the following discussion is for purposes of illustration and analogous techniques may be employed with other polymers.
• PEG first of all, various functionalized polyethylene glycols have been used effectively in fields such as protein modification (see Abuchowski et al., Enzymes as Drugs, John Wiley & Sons: New York, N.Y.
• Activated forms of PEG including multifunctionally activated PEG, are commercially available, and are also easily prepared using known methods. For example, see Chapter 22 of Poly( ethylene Glycol) Chemistry: Biotechnical and Biomedical Applications, J. Milton Harris, ed., Plenum Press, NY (1992); and Shearwater Polymers, Inc. Catalog, Polyethylene Glycol Derivatives,
• FIGS. 1 to 10 of U.S. Patent 5,874,500 structures for some specific, tetrafunctionally activated forms of PEG are shown in FIGS. 1 to 10 of U.S. Patent 5,874,500, as are generalized reaction products obtained by reacting the activated PEGs with multi-amino
• PEGs i.e., a PEG with two or more primary amino groups.
• the activated PEGs illustrated have a pentaerythritol (2,2-bis(hydroxymethyl)-1 ,3-propanediol) core.
• Such activated PEGs are readily prepared by conversion of the exposed hydroxyl groups in the PEGylated polyol
• the crosslinkable compositions of the invention can also include hydrophobic polymers, although for most uses hydrophilic polymers are preferred.
• Polylactic acid and polyglycolic acid are examples of two hydrophobic polymers that can be used. With other hydrophobic polymers, only short-chain oligomers should be used, containing at most about 14 carbon atoms, to avoid solubility-related problems during reaction.
• the molecular core of one or more of the crosslinkable components can also be a low molecular weight compound, i.e., a C 2 -C ⁇ hydrocarbyl group containing zero to 2 heteroatoms selected from N, O, S and combinations thereof.
• a molecular core can be substituted with nucleophilic groups or with electrophilic groups.
• the component may be, for example, ethylenediamine (H 2 N-CH 2 CH 2 -NH 2 ), tetramethylenediamine (H 2 N-(CH 4 )-NH 2 ), pentamethylenediamine (cadaverine) (H 2 N-(CH 5 )-NH 2 ), hexamethylenediamine (H 2 N-(CH 6 )-NH 2 ), bis(2-aminoethyl)amine (HN-[CH 2 CH 2 -NH 2 J 2 ), or tris(2- aminoethyl)amine (N-[CH 2 CH 2 -NH 2 ] 3 ).
• ethylenediamine H 2 N-CH 2 CH 2 -NH 2
• tetramethylenediamine H 2 N-(CH 4 )-NH 2
• pentamethylenediamine cadaverine
• H 2 N-(CH 5 )-NH 2 hexamethylenediamine
• H 2 N-(CH 6 )-NH 2 bis(2-amin
• Low molecular weight diols and polyols include trimethylolpropane, di(trimethylol propane), pentaerythritol, and diglycerol, all of which require activation with a base in order to facilitate their reaction as nucleophiles.
• Such diols and polyols may also be functionalized to provide di- and poly-carboxylic acids, functional groups that are, as noted earlier herein, also useful as nucleophiles under certain conditions.
• Polyacids for use in the present compositions include, without limitation, trimethylolpropane-based tricarboxylic acid, di(trimethylol propane)-based tetracarboxylic acid, heptanedioic acid, octanedioic acid (suberic acid), and hexadecanedioic acid (thapsic acid), all of which are commercially available and/or readily synthesized using known techniques.
• Low molecular weight di- and poly-electrophiles include, for example, disuccinimidyl suberate (DSS), bis(sulfosuccinimidyl) suberate (BS 3 ), dithiobis(succinimidylpropionate) (DSP), bis(2-succinimidooxycarbonyloxy) ethyl sulfone (BSOCOES), and 3,3'-dithiobis(sulfosuccinimidylpropionate (DTSPP), and their analogs and derivatives.
• DSS disuccinimidyl suberate
• BS 3 bis(sulfosuccinimidyl) suberate
• DSP dithiobis(succinimidylpropionate)
• BSOCOES bis(2-succinimidooxycarbonyloxy) ethyl sulfone
• DTSPP 3,3'-dithiobis(sulfosuccin
• di- and poly- electrophiles can also be synthesized from di- and polyacids, for example by reaction with an appropriate molar amount of N-hydroxysuccinimide in the presence of DCC.
• Polyols such as trimethylolpropane and di(trimethylol propane) can be converted to carboxylic acid form using various known techniques, then further derivatized by reaction with NHS in the presence of DCC to produce trifunctionally and tetrafunctionally activated polymers.
• Suitable delivery systems for the homogeneous dry powder composition (containing at least two crosslinkable polymers) and the two buffer solutions may involve a multi-compartment spray device, where one or more compartments contains the powder and one or more compartments contain the buffer solutions needed to provide for the aqueous environment, so that the composition is exposed to the aqueous environment as it leaves the compartment.
• a multi-compartment spray device where one or more compartments contains the powder and one or more compartments contain the buffer solutions needed to provide for the aqueous environment, so that the composition is exposed to the aqueous environment as it leaves the compartment.
• Many devices that are adapted for delivery of multi-component tissue sealants/hemostatic agents are well known in the art and can also be used in the practice of the present invention.
• the composition can be delivered using any type of controllable extrusion system, or it can be delivered manually in the form of a dry powder, and exposed to the aqueous environment at the site of administration.
• the homogeneous dry powder composition and the two buffer solutions may be conveniently formed under aseptic conditions by placing each of the three ingredients (dry powder, acidic buffer solution and basic buffer solution) into separate syringe barrels.
• the composition, first buffer solution and second buffer solution can be housed separately in a multiple-compartment syringe system having a multiple barrels, a mixing head, and an exit orifice.
• the first buffer solution can be added to the barrel housing the composition to dissolve the composition and form a homogeneous solution, which is then extruded into the mixing head.
• the second buffer solution can be simultaneously extruded into the mixing head.
• the resulting composition can then be extruded through the orifice onto a surface.
• the syringe barrels holding the dry powder and the basic buffer may be part of a dual-syringe system, e.g., a double barrel syringe as described in U.S. Patent 4,359,049 to Redl et al.
• the acid buffer can be added to the syringe barrel that also holds the dry powder, so as to produce the homogeneous solution.
• the acid buffer may be added (e.g., injected) into the syringe barrel holding the dry powder to thereby produce a homogeneous solution of the first and second components. This homogeneous solution can then be extruded into a mixing head, while the basic buffer is simultaneously extruded into the mixing head.
• the homogeneous solution and the basic buffer are mixed together to thereby form a reactive mixture.
• the reactive mixture is extruded through an orifice and onto a surface (e.g., tissue), where a film is formed, which can function as a sealant or a barrier, or the like.
• the reactive mixture begins forming a three-dimensional matrix immediately upon being formed by the mixing of the homogeneous solution and the basic buffer in the mixing head. Accordingly, the reactive mixture is preferably extruded from the mixing head onto the tissue very quickly after it is formed so that the three-dimensional matrix forms on, and is able to adhere to, the tissue.
• Other systems for combining two reactive liquids are well known in the art, and include the systems described in U.S. Patent Nos.
• the electrophilic component or components are generally stored and used in sterile, dry form to prevent hydrolysis.
• Processes for preparing synthetic hydrophilic polymers containing multiple electrophilic groups in sterile, dry form are set forth in commonly assigned U.S. Patent No. 5,643,464 to Rhee et al.
• the dry synthetic polymer may be compression molded into a thin sheet or membrane, which can then be sterilized using gamma or, preferably, e-beam irradiation. The resulting dry membrane or sheet can be cut to the desired size or chopped into smaller size particulates.
• Components containing multiple nucleophilic groups are generally not water-reactive and can therefore be stored either dry or in aqueous solution. If stored as a dry, particulate, solid, the various components of the crosslinkable composition may be blended and stored in a single container. Admixture of all components with water, saline, or other aqueous media should not occur until immediately prior to use.
• the crosslinking components can be mixed together in a single aqueous medium in which they are both unreactive, i.e., such as in a low pH buffer. Thereafter, they can be sprayed onto the targeted tissue site along with a high pH buffer, after which they will rapidly react and form a gel.
• Suitable liquid media for storage of crosslinkable compositions include aqueous buffer solutions such as monobasic sodium phosphate/dibasic sodium phosphate, sodium carbonate/sodium bicarbonate, glutamate or acetate, at a concentration of 0.5 to 300 mM.
• a sulfhydryl-reactive component such as PEG substituted with maleimido groups or succinimidyl esters is prepared in water or a dilute buffer, with a pH of between around 5 to 6.
• Buffers with pKs between about 8 and 10.5 for preparing a polysulfhydryl component such as sulfhydryl-PEG are useful to achieve fast gelation time of compositions containing mixtures of sulfhydryl-PEG and SG-PEG.
• These include carbonate, borate and AMPSO (3-[(1 ,1-dimethyl-2- hydroxyethyl)amino]2-hydroxy-propane-sulfonic acid).
• a pH of around 5 to 9 is preferred for the liquid medium used to prepare the sulfhydryl PEG.
• the polymer composition may include collagen in combination with fibrinogen and/or thrombin.
• an aqueous composition may include a fibrinogen and FXIII, particularly plasma, collagen in an amount sufficient to thicken the composition, thrombin in an amount sufficient to catalyze polymerization of fibrinogen present in the composition, and Ca 2+ and, optionally, an antifibrinolytic agent in amount sufficient to retard degradation of the resulting adhesive clot.
• the composition may be formulated as a two-part composition that may be mixed together just prior to use, in which fibrinogen/FXIII and collagen constitute the first component, and thrombin together with an antifibrinolytic agent, and Ca 2+ constitute the second component.
• Plasma which provides a source of fibrinogen, may be obtained from the patient for which the composition is to be delivered.
• the plasma can be used "as is” after standard preparation which includes centrifuging out cellular components of blood.
• the plasma can be further processed to concentrate the fibrinogen to prepare a plasma cryoprecipitate.
• the plasma cryoprecipitate can be prepared by freezing the plasma for at least about an hour at about -20 °C, and then storing the frozen plasma overnight at about 4 °C.
• the thawed plasma is centrifuged and the plasma cryoprecipitate is harvested by removing approximately four-fifths of the plasma to provide a cryoprecipitate comprising the remaining one-fifth of the plasma.
• Other fibrinogen/FXIII preparations may be used, such as cryoprecipitate, patient autologous fibrin sealant, fibrinogen analogs or other single donor or commercial fibrin sealant materials.
• Approximately 0.5 ml to about 1.0 ml of either the plasma or the plasma-cryoprecipitate provides about 1 to 2 ml of adhesive composition which is sufficient for use in middle ear surgery.
• Plasma proteins may or may not be present in the fibrinogen/FXII separation due to wide variations in the formulations and methods to derive them.
• Collagen preferably hypoallergenic collagen, is present in the composition in an amount sufficient to thicken the composition and augment the cohesive properties of the preparation.
• the collagen may be atelopeptide collagen or telopeptide collagen, e.g., native collagen.
• the collagen augments the fibrin by acting as a macromolecular lattice work or scaffold to which the fibrin network adsorbs.
• the form of collagen which is employed may be described as at least "near native" in its structural characteristics. It may be further characterized as resulting in insoluble fibers at a pH above 5; unless crosslinked or as part of a complex composition, e.g., bone, it will generally consist of a minor amount by weight of fibers with diameters greater than 50 nm, usually from about 1 to 25 volume % and there will be substantially little, if any, change in the helical structure of the fibrils. In addition, the collagen composition must be able to enhance gelation in the surgical adhesion composition.
• ZYDERM Collagen Implant has a fibrillar diameter distribution consisting of 5 to 10 nm diameter fibers at 90% volume content and the remaining 10% with greater than about 50 nm diameter fibers.
• ZCI is available as a fibrillar slurry and solution in phosphate buffered isotonic saline, pH 7.2, and is injectable with fine gauge needles.
• cross-linked collagen available as ZYPLAST may be employed.
• ZYPLAST is essentially an exogenously crosslinked (glutaraldehyde) version of ZCI. The material has a somewhat higher content of greater than about 50 nm diameter fibrils and remains insoluble over a wide pH range.
• Crosslinking has the effect of mimicking in vivo endogenous crosslinking found in many tissues.
• Thrombin acts as a catalyst for fibrinogen to provide fibrin, an insoluble polymer and is present in the composition in an amount sufficient to catalyze polymerization of fibrinogen present in the patient plasma.
• Thrombin also activates FXIII, a plasma protein that catalyzes covalent crosslinks in fibrin, rendering the resultant clot insoluble.
• the thrombin is present in the adhesive composition in concentration of from about 0.01 to about 1000 or greater NIH units (NIHu) of activity, usually about i to about 500 NIHu, most usually about 200 to about 500 NIHu.
• the thrombin can be from a variety of host animal sources, conveniently bovine. Thrombin is commercially available from a variety of sources including Parke-Davis, usually lyophilized with buffer salts and stabilizers in vials which provide thrombin activity ranging from about 1000 NIHu to 10,000 NIHu.
• the thrombin is usually prepared by reconstituting the powder by the addition of either sterile distilled water or isotonic saline. Alternately, thrombin analogs or reptile-sourced coagulants may be used.
• the composition may additionally comprise an effective amount of an antifibrinolytic agent to enhance the integrity of the glue clot as the healing processes occur.
• a number of antifibrinolytic agents are well known and include aprotinin, C1-esterase inhibitor and ⁇ -amino-n-caproic acid (EACA).
• ⁇ - amino-n-caproic acid the only antifibrinolytic agent approved by the FDA, is effective at a concentration of from about 5 mg/ml to about 40 mg/ml of the final adhesive composition, more usually from about 20 to about 30 mg/ml.
• EACA is commercially available as a solution having a concentration of about 250 mg/ml. Conveniently, the commercial solution is diluted with distilled water to provide a solution of the desired concentration. That solution is desirably used to reconstitute lyophilized thrombin to the desired thrombin concentration.
• in situ forming materials based on the crosslinking of proteins are described, e.g., in U.S. Patent Nos. RE38158; 4,839,345; 5,514,379, 5,583,114; 6,458,147; 6,371 ,975; 5,290,552; 6,096,309; U.S. Patent Application Publication Nos. 2002/0161399; 2001/0018598 and PCT Publication Nos. WO 03/090683; WO 01/45761 ; WO 99/66964 and WO 96/03159).
• the therapeutic agent is released from a crosslinked matrix formed, at least in part, from a self-reactive compound.
• a self-reactive compound comprises a core substituted with a minimum of three reactive groups.
• the reactive groups may be directed attached to the core of the compound, or the reactive groups may be indirectly attached to the compound's core, e.g., the reactive groups are joined to the core through one or more linking groups.
• Each of the three reactive groups that are necessarily present in a self-reactive compound can undergo a bond-forming reaction with at least one of the remaining two reactive groups. For clarity it is mentioned that when these compounds react to form a crosslinked matrix, it will most often happen that reactive groups on one compound will reactive with reactive groups on another compound.
• the term "self-reactive” is not intended to mean that each self-reactive compound necessarily reacts with itself, but rather that when a plurality of identical self-reactive compounds are in combination and undergo a crosslinking reaction, then these compounds will react with one another to form the matrix.
• the compounds are "self-reactive” in the sense that they can react with other compounds having the identical chemical structure as themselves.
• the self-reactive compound comprises at least four components: a core and three reactive groups.
• the self-reactive compound can be characterized by the formula (I), where R is the core, the reactive groups are represented by X 1 , X 2 and X 3 , and a linker (L) is optionally present between the core and a functional group.
• the core R is a polyvalent moiety having attachment to at least three groups (i.e., it is at least trivalent) and may be, or may contain, for example, a hydrophilic polymer, a hydrophobic polymer, an amphiphilic polymer, a C 2 - 14 hydrocarbyl, or a C 2- ⁇ 4 hydrocarbyl which is heteroatom- containing.
• the linking groups L 1 , L 2 , and L 3 may be the same or different.
• the designators p, q and r are either 0 (when no linker is present) or 1 (when a linker is present).
• the reactive groups X 1 , X 2 and X 3 may be the same or different.
• each of these reactive groups reacts with at least one other reactive group to form a three-dimensional matrix. Therefore X 1 can react with X 2 and/or X 3 , X 2 can react with X 1 and/or X 3 , X 3 can react with X 1 and/or X 2 and so forth.
• a trivalent core will be directly or indirectly bonded to three functional groups, a tetravalent core will be directly or indirectly bonded to four functional groups, etc.
• Each side chain typically has one reactive group.
• the invention also encompasses self-reactive compounds where the side chains contain more than one reactive group.
• the self-reactive compound has the formula (II):
• the compound is essentially non-reactive in an initial environment but is rendered reactive upon exposure to a modification in the initial environment that provides a modified environment such that a plurality of the self-reactive compounds inter-react in the modified environment to form a three-dimensional matrix.
• R is a hydrophilic polymer.
• X' is a nucleophilic group and Y' is an electrophilic group.
• the following self-reactive compound is one example of a compound of formula (II):
• R has the formula:
• the self-reactive compounds of the invention are readily synthesized by techniques that are well known in the art. An exemplary synthesis is set forth below:
• the reactive groups are selected so that the compound is essentially non-reactive in an initial environment.
• the compound Upon exposure to a specific modification in the initial environment, providing a modified environment, the compound is rendered reactive and a plurality of self-reactive compounds are then able to inter-react in the modified environment to form a three-dimensional matrix.
• modification in the initial environment include the addition of an aqueous medium, a change in pH, exposure to ultraviolet radiation, a change in temperature, or contact with a redox initiator.
• the core and reactive groups can also be selected so as to provide a compound that has one of more of the following features: are biocompatible, are non-immunogenic, and do not leave any toxic, inflammatory or immunogenic reaction products at the site of administration.
• the core and reactive groups can also be selected so as to provide a resulting matrix that has one or more of these features.
• substantially immediately or immediately upon exposure to the modified environment the self-reactive compounds inter-react form a three-dimensional matrix.
• the term "substantially immediately” is intended to mean within less than five minutes, preferably within less than two minutes, and the term “immediately” is intended to mean within less than one minute, preferably within less than 30 seconds.
• the self-reactive compound and resulting matrix are not subject to enzymatic cleavage by matrix metalloproteinases such as collagenase, and are therefore not readily degradable in vivo.
• the self-reactive compound may be readily tailored, in terms of the selection and quantity of each component, to enhance certain properties, e.g., compression strength, swellability, tack, hydrophilicity, optical clarity, and the like.
• R is a hydrophilic polymer.
• X is a nucleophilic group
• Y is an electrophilic group
• Z is either an electrophilic or a nucleophilic group. Additional embodiments are detailed below.
• a higher degree of inter-reaction e.g., crosslinking
• n be an integer from 2-12.
• the compounds may be the same or different.
• the self-reactive compound Prior to use, the self-reactive compound is stored in an initial environment that insures that the compound remain essentially non-reactive until use. Upon modification of this environment, the compound is rendered reactive and a plurality of compounds will then inter-react to form the desired matrix.
• the initial environment, as well as the modified environment, is thus determined by the nature of the reactive groups involved.
• the number of reactive groups can be the same or different. However, in one embodiment of the invention, the number of reactive groups are approximately equal.
• the term "approximately” refers to a 2:1 to 1 :2 ratio of moles of one reactive group to moles of a different reactive groups. A 1 :1 :1 molar ratio of reactive groups is generally preferred.
• the concentration of the self-reactive compounds in the modified environment when liquid in nature, will be in the range of about 1 to 50 wt%, generally about 2 to 40 wt%.
• the preferred concentration of the compound in the liquid will depend on a number of factors, including the type of compound (i.e., type of molecular core and reactive groups), its molecular weight, and the end use of the resulting three-dimensional matrix. For example, use of higher concentrations of the compounds, or using highly functionalized compounds, will result in the formation of a more tightly crosslinked network, producing a stiffer, more robust gel.
• compositions intended for use in tissue augmentation will generally employ concentrations of self-reactive compounds that fall toward the higher end of the preferred concentration range.
• Compositions intended for use as bioadhesives or in adhesion prevention do not need to be as firm and may therefore contain lower concentrations of the self-reactive compounds.
• the reactive groups are electrophilic and nucleophilic groups, which undergo a nucleophilic substitution reaction, a nucleophilic addition reaction, or both.
• electrophilic refers to a reactive group that is susceptible to nucleophilic attack, i.e., susceptible to reaction with an incoming nucleophilic group.
• Electrophilic groups herein are positively charged or electron-deficient, typically electron- deficient.
• nucleophilic refers to a reactive group that is electron rich, has an unshared pair of electrons acting as a reactive site, and reacts with a positively charged or electron-deficient site.
• the modification in the initial environment comprises the addition of an aqueous medium and/or a change in pH.
• X1 also referred to herein as
• X can be a nucleophilic group and X2 (also referred to herein as Y) can be an electrophilic group or vice versa, and X3 (also referred to herein as Z) can be either an electrophilic or a nucleophilic group.
• X may be virtually any nucleophilic group, so long as reaction can occur with the electrophilic group Y and also with Z, when Z is electrophilic (ZEL)-
• Y may be virtually any electrophilic group, so long as reaction can take place with X and also with Z when Z is nucleophilic (ZNU)-
• ZEL electrophilic
• ZNU nucleophilic
• the reactions between X and Y, and between either X and ZEL or Y and ZNU are complete in under 60 minutes, preferably under 30 minutes. Most preferably, the reaction occurs in about 5 to 15 minutes or less.
• nucleophilic groups suitable as X or F ⁇ NU include, but are not limited to: -NH 2 , -NHR 1 , -N(R 1 ) 2 , -SH, -OH, -COOH, -C 6 H 4 -OH, -H, -PH 2 , -PHR 1 , -P(R 1 ) 2 , -NH-NH 2 , -CO-NH-NH 2 , -C 5 H 4 N, etc.
• R 1 is a hydrocarbyl group and each R1 may be the same or different.
• R 1 is typically alkyl or monocyclic aryl, preferably alkyl, and most preferably lower alkyl.
• Organometallic moieties are also useful nucleophilic groups for the purposes of the invention, particularly those that act as carbanion donors. Examples of organometallic moieties include: Grignard functionalities -R 2 MgHal wherein R 2 is a carbon atom (substituted or unsubstituted), and Hal is halo, typically bromo, iodo or chloro, preferably bromo; and lithium-containing functionalities, typically alkyllithium groups; sodium-containing functionalities.
• nucleophilic groups must be activated with a base so as to be capable of reaction with an electrophilic group.
• the compound when there are nucleophilic sulfhydryl and hydroxyl groups in the self-reactive compound, the compound must be admixed with an aqueous base in order to remove a proton and provide an -S " or -O " species to enable reaction with the electrophilic group.
• a non- nucleophilic base is preferred.
• the base may be present as a component of a buffer solution. Suitable bases and corresponding crosslinking reactions are described herein.
• electrophilic groups provided on the self-reactive compound must be made so that reaction is possible with the specific nucleophilic groups.
• the Y and any Z E L groups are selected so as to react with amino groups.
• the corresponding electrophilic groups are sulfhydryl-reactive groups, and the like.
• the amine-reactive groups contain an electrophilically reactive carbonyl group susceptible to nucleophilic attack by a primary or secondary amine, for example the carboxylic acid esters and aldehydes noted above, as well as carboxyl groups (-COOH). Since a carboxylic acid group per se is not susceptible to reaction with a nucleophilic amine, components containing carboxylic acid groups must be activated so as to be amine-reactive. Activation may be accomplished in a variety of ways, but often involves reaction with a suitable hydroxyl-containing compound in the presence of a dehydrating agent such as dicyclohexylcarbodiimide (DCC) or dicyclohexylurea (DHU).
• DCC dicyclohexylcarbodiimide
• DHU dicyclohexylurea
• a carboxylic acid can be reacted with an alkoxy-substituted N-hydroxysuccinimide or N-hydroxysulfosuccinimide in the presence of DCC to form reactive electrophilic groups, the N-hydroxysuccinimide ester and the N- hydroxysulfosuccinimide ester, respectively.
• Carboxylic acids may also be activated by reaction with an acyl halide such as an acyl chloride (e.g., acetyl chloride), to provide a reactive anhydride group.
• a carboxylic acid may be converted to an acid chloride group using, e.g., thionyl chloride or an acyl chloride capable of an exchange reaction.
• the amine-reactive groups are selected from succinimidyl ester (-0(CO)-N(COCH 2 ) 2 ), sulfosuccinimidyl ester (-0(CO)-N(COCH 2 ) 2 -S(0) 2 OH), maleimido (-N(COCH) 2 ), epoxy, isocyanato, thioisocyanato, and ethenesulfonyl.
• the electrophilic groups present on Y and ZEL are groups that react with a sulfhydryl moiety.
• Such reactive groups include those that form thioester linkages upon reaction with a sulfhydryl group, such as those described in WO 00/62827 to Wallace et al.
• sulfhydryl reactive groups include, but are not limited to: mixed anhydrides; ester derivatives of phosphorus; ester derivatives of p-nitrophenol, p-nitrothiophenol and pentafluorophenol; esters of substituted hydroxylamines, including N-hydroxyphthalimide esters, N-hydroxysuccinimide esters, N- hydroxysulfosuccinimide esters, and N-hydroxyglutarimide esters; esters of 1- hydroxybenzotriazole; 3-hydroxy-3,4-dihydro-benzotriazin-4-one; 3-hydroxy- 3,4-dihydro-quinazoline-4-one; carbonylimidazole derivatives; acid chlorides; ketenes; and isocyanates.
• auxiliary reagents can also be used to facilitate bond formation, e.g., 1 -ethyl-3-[3- dimethylaminopropyl]carbodiimide can be used to facilitate coupling of sulfhydryl groups to carboxyl-containing groups.
• various other sulfhydryl reactive functionalities can be utilized that form other types of linkages. For example, compounds that contain methyl imidate derivatives form imido-thioester linkages with sulfhydryl groups.
• sulfhydryl reactive groups can be employed that form disulfide bonds with sulfhydryl groups; such groups generally have the structure -S-S-Ar where Ar is a substituted or unsubstituted nitrogen-containing heteroaromatic moiety or a non-heterocyclic aromatic group substituted with an electron- withdrawing moiety, such that Ar may be, for example, 4-pyridinyl, o- nitrophenyl, m-nitrophenyl, p-nitro phenyl, 2,4-dinitrophenyl, 2-nitro-4-benzoic acid, 2-nitro-4-pyridinyl, etc.
• auxiliary reagents i.e., mild oxidizing agents such as hydrogen peroxide
• sulfhydryl reactive groups forms thioether bonds with sulfhydryl groups.
• groups include, inter alia, maleimido, substituted maleimido, haloalkyl, epoxy, imino, and aziridino, as well as olefins (including conjugated olefins) such as ethenesulfonyl, etheneimino, acrylate, methacrylate, and ⁇ , ⁇ -unsaturated aldehydes and ketones.
• the electrophilic functional groups on the remaining component(s) must react with hydroxyl groups.
• the hydroxyl group may be activated as described above with respect to carboxylic acid groups, or it may react directly in the presence of base with a sufficiently reactive electrophilic group such as an epoxide group, an aziridine group, an acyl halide, an anhydride, and so forth.
• X is an organometallic nucleophilic group such as a Grignard functionality or an alkyllithium group
• suitable electrophilic functional groups for reaction therewith are those containing carbonyl groups, including, by way of example, ketones and aldehydes.
• certain functional groups can react as nucleophilic or as electrophilic groups, depending on the selected reaction partner and/or the reaction conditions.
• a carboxylic acid group can act as a nucleophilic group in the presence of a fairly strong base, but generally acts as an electrophilic group allowing nucleophilic attack at the carbonyl carbon and concomitant replacement of the hydroxyl group with the incoming nucleophilic group.
• the initial environment typically can be dry and sterile. Since electrophilic groups react with water, storage in sterile, dry form will prevent hydrolysis.
• the dry synthetic polymer may be compression molded into a thin sheet or membrane, which can then be sterilized using gamma or e- beam irradiation. The resulting dry membrane or sheet can be cut to the desired size or chopped into smaller size particulates.
• the modification of a dry initial environment will typically comprise the addition of an aqueous medium.
• the initial environment can be an aqueous medium such as in a low pH buffer, i.e., having a pH less than about 6.0, in which both electrophilic and nucleophilic groups are non-reactive.
• aqueous buffer solutions such as monobasic sodium phosphate/dibasic sodium phosphate, sodium carbonate/sodium bicarbonate, glutamate or acetate, at a concentration of 0.5 to 300 mM.
• Modification of an initial low pH aqueous environment will typically comprise increasing the pH to at least pH 7.0, more preferably increasing the pH to at least pH 9.5.
• the modification of a dry initial environment comprises dissolving the self-reactive compound in a first buffer solution having a pH within the range of about 1.0 to 5.5 to form a homogeneous solution, and (ii) adding a second buffer solution having a pH within the range of about 6.0 to 11.0 to the homogeneous solution.
• the buffer solutions are aqueous and can be any pharmaceutically acceptable basic or acid composition.
• the term "buffer" is used in a general sense to refer to an acidic or basic aqueous solution, where the solution may or may not be functioning to provide a buffering effect (i.e., resistance to change in pH upon addition of acid or base) in the compositions of the present invention.
• the self-reactive compound can be in the form of a homogeneous dry powder.
• This powder is then combined with a buffer solution having a pH within the range of about 1.0 to 5.5 to form a homogeneous acidic aqueous solution, and this solution is then combined with a buffer solution having a pH within the range of about 6.0 to 11.0 to form a reactive solution.
• 0.375 grams of the dry powder can be combined with 0.75 grams of the acid buffer to provide, after mixing, a homogeneous solution, where this solution is combined with 1.1 grams of the basic buffer to provide a reactive mixture that substantially immediately forms a three-dimensional matrix.
• Acidic buffer solutions having a pH within the range of about 1.0 to 5.5 include by way of illustration and not limitation, solutions of: citric acid, hydrochloric acid, phosphoric acid, sulfuric acid, AMPSO (3-[(1 ,1-dimethyl-2- hydroxyethyl)amino]2-hydroxy-propane-sulfonic acid), acetic acid, lactic acid, and combinations thereof.
• the acidic buffer solution is a solution of citric acid, hydrochloric acid, phosphoric acid, sulfuric acid, and combinations thereof.
• the acidic buffer preferably has a pH such that it retards the reactivity of the nucleophilic groups on the core.
• the acidic buffer is an acidic solution that, when contacted with nucleophilic groups, renders those nucleophilic groups relatively non-nucleophilic.
• An exemplary acidic buffer is a solution of hydrochloric acid, having a concentration of about 6.3 mM and a pH in the range of 2.1 to 2.3. This buffer may be prepared by combining concentrated hydrochloric acid with water, i.e., by diluting concentrated hydrochloric acid with water.
• this buffer A may also be conveniently prepared by diluting 1.23 grams of concentrated hydrochloric acid to a volume of 2 liters, or diluting 1.84 grams of concentrated hydrochloric acid to a volume to 3 liters, or diluting 2.45 grams of concentrated hydrochloric acid to a volume of 4 liters, or diluting 3.07 grams concentrated hydrochloric acid to a volume of 5 liters, or diluting 3.68 grams of concentrated hydrochloric acid to a volume to 6 liters.
• the concentrated acid is preferably added to water.
• Basic buffer solutions having a pH within the range of about 6.0 to 11.0 include by way of illustration and not limitation, solutions of: glutamate, acetate, carbonate and carbonate salts (e.g., sodium carbonate, sodium carbonate monohydrate and sodium bicarbonate), borate, phosphate and phosphate salts (e.g., monobasic sodium phosphate monohydrate and dibasic sodium phosphate), and combinations thereof.
• the basic buffer solution is a solution of carbonate salts, phosphate salts, and combinations thereof.
• the basic buffer is an aqueous solution that neutralizes the effect of the acidic buffer, when it is added to the homogeneous solution of the compound and first buffer, so that the nucleophilic groups on the core regain their nucleophilic character (that has been masked by the action of the acidic buffer), thus allowing the nucleophilic groups to inter-react with the electrophilic groups on the core.
• An exemplary basic buffer is an aqueous solution of carbonate and phosphate salts. This buffer may be prepared by combining a base solution with a salt solution. The salt solution may be prepared by combining 34.7 g of monobasic sodium phosphate monohydrate, 49.3 g of sodium carbonate monohydrate, and sufficient water to provide a solution volume of 2 liter.
• a 6 liter solution may be prepared by combining 104.0 g of monobasic sodium phosphate monohydrate, 147.94 g of sodium carbonate monohydrate, and sufficient water to provide 6 liter of the salt solution.
• the basic buffer may be prepared by combining 7.2 g of sodium hydroxide with 180.0 g of water.
• the basic buffer is typically prepared by adding the base solution as needed to the salt solution, ultimately to provide a mixture having the desired pH, e.g., a pH of 9.65 to 9.75.
• the basic species present in the basic buffer should be sufficiently basic to neutralize the acidity provided by the acidic buffer, but should not be so nucleophilic itself that it will react substantially with the electrophilic groups on the core.
• a three-dimensional matrix of the present invention may combine an admixture of the self-reactive compound with a first, acidic, buffer (e.g., an acid solution, e.g., a dilute hydrochloric acid solution) to form a homogeneous solution.
• a first, acidic, buffer e.g., an acid solution, e.g., a dilute hydrochloric acid solution
• This homogeneous solution is mixed with a second, basic, buffer (e.g., a basic solution, e.g., an aqueous solution containing phosphate and carbonate salts) whereupon the reactive groups on the core of the self-reactive compound substantially immediately inter-react with one another to form a three- dimensional matrix.
• the reactive groups are vinyl groups such as styrene derivatives, which undergo a radical polymerization upon initiation with a redox initiator.
• redox refers to a reactive group that is susceptible to oxidation-reduction activation.
• vinyl refers to a reactive group that is activated by a redox initiator, and forms a radical upon reaction.
• X, Y and Z can be the same or different vinyl groups, for example, methacrylic groups.
• the initial environment typically will be an aqueous environment. The modification of the initial environment involves the addition of a redox initiator.
• the reactive groups undergo an oxidative coupling reaction.
• X, Y and Z can be a halo group such as chloro, with an adjacent electron-withdrawing group on the halogen- bearing carbon (e.g., on the "L" linking group).
• exemplary electron-withdrawing groups include nitro, aryl, and so forth.
• the modification in the initial environment comprises a change in pH.
• a base such as KOH
• the self-reactive compounds then undergo a de-hydro, chloro coupling reaction, forming a double bond between the carbon atoms, as illustrated below:
• the initial environment typically can be can be dry and sterile, or a non-basic medium.
• the modification of the initial environment will typically comprise the addition of a base.
• the reactive groups are photoinitiated groups.
• the modification in the initial environment comprises exposure to ultraviolet radiation.
• X can be an azide (-N 3 ) group and Y can be an alkyl group such as -CH(CH 3 ) 2 or vice versa. Exposure to ultraviolet radiation will then form a bond between the groups to provide for the following linkage: -NH-C(CH 3 ) 2 -CH 2 -.
• X can be a benzophenone (-(C 6 H 4 )-C(0)-(C 6 H5)) group and Y can be an alkyl group such as -CH(CH 3 ) 2 or vice versa. Exposure to ultraviolet radiation will then form a bond between the groups to provide for the following linkage:
• the initial environment typically will be in an ultraviolet radiation- shielded environment. This can be for example, storage within a container that is impermeable to ultraviolet radiation.
• the modification of the initial environment will typically comprise exposure to ultraviolet radiation.
• the reactive groups are temperature-sensitive groups, which undergo a thermochemical reaction.
• the modification in the initial environment thus comprises a change in temperature.
• temperature-sensitive refers to a reactive group that is chemically inert at one temperature or temperature range and reactive at a different temperature or temperature range.
• X, Y, and Z are the same or different vinyl groups.
• the initial environment typically will be within the range of about 10 to 30°C.
• the modification of the initial environment will typically comprise changing the temperature to within the range of about 20 to 40°C.
• Linking Groups The reactive groups may be directly attached to the core, or they may be indirectly attached through a linking group, with longer linking groups also termed "chain extenders.”
• chain extenders In the formula (I) shown above, the optional linker groups are represented by L 1 , L 2 , and L 3 , wherein the linking groups are present when p, q and r are equal to 1.
• Suitable linking groups are well known in the art. See, for example, WO 97/22371 to Rhee et al. Linking groups are useful to avoid steric hindrance problems that can sometimes associated with the formation of direct linkages between molecules. Linking groups may additionally be used to link several self-reactive compounds together to make larger molecules.
• a linking group can be used to alter the degradative properties of the compositions after administration and resultant gel formation.
• linking groups can be used to promote hydrolysis, to discourage hydrolysis, or to provide a site for enzymatic degradation.
• Examples of linking groups that provide hydrolyzable sites include, inter alia: ester linkages; anhydride linkages, such as those obtained by incorporation of glutarate and succinate; ortho ester linkages; ortho carbonate linkages such as trimethylene carbonate; amide linkages; phosphoester linkages; ⁇ -hydroxy acid linkages, such as those obtained by incorporation of lactic acid and glycolic acid; lactone-based linkages, such as those obtained by incorporation of caprolactone, valerolactone, ⁇ -butyrolactone and p-dioxanone; and amide linkages such as in a dimeric, oligomeric, or poly(amino acid) segment.
• non-degradable linking groups include succinimide, propionic acid and carboxymethylate linkages. See, for example, WO 99/07417 to Coury et al.
• enzymatically degradable linkages include Leu- Gly-Pro-Ala, which is degraded by collagenase; and Gly-Pro-Lys, which is degraded by plasmin.
• Linking groups can also be included to enhance or suppress the reactivity of the various reactive groups. For example, electron-withdrawing groups within one or two carbons of a sulfhydryl group would be expected to diminish its effectiveness in coupling, due to a lowering of nucleophilicity. Carbon-carbon double bonds and carbonyl groups will also have such an effect.
• x is generally in the range of 1 to about 10;
• R 2 is generally hydrocarbyl, typically alkyl or aryl, preferably alkyl, and most preferably lower alkyl; and
• R 3 is hydrocarbylene, heteroatom-containing hydrocarbylene, substituted hydrocarbylene, or substituted heteroatom- containing hydrocarbylene) typically alkylene or arylene (again, optionally substituted and/or containing a heteroatom), preferably lower alkylene (e.g., methylene, ethylene, n-propylene, n-butylene, etc.), phenylene, or amidoalkylene (e.g., -(CO)-NH-CH 2 ).
• lower alkylene e.g., methylene, ethylene, n-propylene, n-butylene, etc.
• phenylene or amidoalkylene (e.g., -(CO)-NH-CH 2 ).
• linking groups are as follows. If a higher molecular weight self-reactive compound is to be used, it will preferably have biodegradable linkages as described above, so that fragments larger than 20,000 mol. wt. are not generated during resorption in the body. In addition, to promote water miscibility and/or solubility, it may be desired to add sufficient electric charge or hydrophilicity. Hydrophilic groups can be easily introduced using known chemical synthesis, so long as they do not give rise to unwanted swelling or an undesirable decrease in compressive strength. In particular, polyalkoxy segments may weaken gel strength.
• each self-reactive compound is comprised of the molecular structure to which the reactive groups are bound.
• the molecular core can a polymer, which includes synthetic polymers and naturally occurring polymers.
• the core is a polymer containing repeating monomer units.
• the polymers can be hydrophilic, hydrophobic, or amphiphilic.
• the molecular core can also be a low molecular weight components such as a C- 2 - 14 hydrocarbyl or a heteroatom-containing C 2- ⁇ 4 hydrocarbyl.
• the heteroatom-containing C 2 - 14 hydrocarbyl can have 1 or 2 heteroatoms selected from N, O and S.
• the self-reactive compound comprises a molecular core of a synthetic hydrophilic polymer.
• Hydrophilic polymer refers to a polymer having an average molecular weight and composition that naturally renders, or is selected to render the polymer as a whole
• hydrophilic Preferred polymers are highly pure or are purified to a highly pure state such that the polymer is or is treated to become pharmaceutically pure. Most hydrophilic polymers can be rendered water soluble by incorporating a sufficient number of oxygen (or less frequently nitrogen) atoms available for forming hydrogen bonds in aqueous solutions. Synthetic hydrophilic polymers may be homopolymers, block copolymers including di-block and tri-block copolymers, random copolymers, or graft copolymers. In addition, the polymer may be linear or branched, and if branched, may be minimally to highly branched, dendrimeric, hyperbranched, or a star polymer.
• the polymer may include biodegradable segments and blocks, either distributed throughout the polymer's molecular structure or present as a single block, as in a block copolymer.
• Biodegradable segments preferably degrade so as to break covalent bonds.
• biodegradable segments are segments that are hydrolyzed in the presence of water and/or enzymatically cleaved in situ.
• Biodegradable segments may be composed of small molecular segments such as ester linkages, anhydride linkages, ortho ester linkages, ortho carbonate linkages, amide linkages, phosphonate linkages, etc. Larger biodegradable "blocks" will generally be composed of oligomeric or polymeric segments incorporated within the hydrophilic polymer.
• Illustrative oligomeric and polymeric segments that are biodegradable include, by way of example, poly(amino acid) segments, poly(orthoester) segments, poly(orthocarbonate) segments, and the like.
• Other biodegradable segments that may form part of the hydrophilic polymer core include polyesters such as polylactide, polyethers such as polyalkylene oxide, polyamides such as a protein, and polyurethanes.
• the core of the self-reactive compound can be a diblock copolymer of tetrafunctionally activated polyethylene glycol and polylactide.
• Synthetic hydrophilic polymers that are useful herein include, but are not limited to: polyalkylene oxides, particularly polyethylene glycol (PEG) and poly( ethylene oxide)-poly(propylene oxide) copolymers, including block and random copolymers; polyols such as glycerol, polyglycerol (PG) and particularly highly branched polyglycerol, propylene glycol; poly(oxyalkylene)-substituted diols, and poly(oxyalkylene)-substituted polyols such as mono-, di- and tri- polyoxyethylated glycerol, mono- and di-polyoxyethylated propylene glycol, and mono- and di-polyoxyethylated trimethylene glycol; polyoxyethylated sorbitol, polyoxyethylated glucose; poly(acrylic acids) and analogs and copolymers thereof, such as polyacrylic acid per se, polymethacrylic acid
• a range of molecular weights indicates that the average molecular weight may be any value between the limits specified, and may include molecules outside those limits.
• a molecular weight range of about 800 to about 20,000 indicates an average molecular weight of at least about 800, ranging up to about 20 kDa.
• Other suitable synthetic hydrophilic polymers include chemically synthesized polypeptides, particularly polynucleophilic polypeptides that have been synthesized to incorporate amino acids containing primary amino groups (such as lysine) and/or amino acids containing thiol groups (such as cysteine). Poly(lysine), a synthetically produced polymer of the amino acid lysine (145 MW), is particularly preferred.
• Poly(lysine)s have been prepared having anywhere from 6 to about 4,000 primary amino groups, corresponding to molecular weights of about 870 to about 580,000.
• Poly(lysine)s for use in the present invention preferably have a molecular weight within the range of about 1 ,000 to about 300,000, more preferably within the range of about 5,000 to about 100,000, and most preferably, within the range of about 8,000 to about 15,000.
• Poly(lysine)s of varying molecular weights are commercially available from Peninsula Laboratories, Inc. (Belmont, Calif.). Although a variety of different synthetic hydrophilic polymers can be used in the present compounds, preferred synthetic hydrophilic polymers are PEG and PG, particularly highly branched PG.
• PEG polystyrene-maleic anhydride
• a particularly preferred synthetic hydrophilic polymer for certain applications is a PEG having a molecular weight within the range of about 100 to about 100,000, although for highly branched PEG, far higher molecular weight polymers can be employed, up to 1 ,000,000 or more, providing that biodegradable sites are incorporated ensuring that all degradation products will have a molecular weight of less than about 30,000.
• the preferred molecular weight is about 1 ,000 to about 20,000, more preferably within the range of about 7,500 to about 20,000.
• the polyethylene glycol has a molecular weight of approximately 10,000.
• Naturally occurring hydrophilic polymers include, but are not limited to: proteins such as collagen, fibronectin, albumins, globulins, fibrinogen, fibrin and thrombin, with collagen particularly preferred; carboxylated polysaccharides such as polymannuronic acid and polygalacturonic acid; aminated polysaccharides, particularly the glycosaminoglycans, e.g., hyaluronic acid, chitin, chondroitin sulfate A, B, or C, keratin sulfate, keratosulfate and heparin; and activated polysaccharides such as dextran and starch derivatives.
• Collagen and glycosaminoglycans are preferred naturally occurring hydrophilic polymers for use herein.
• the term "collagen” as used herein refers to all forms of collagen, including those, which have been processed or otherwise modified.
• collagen from any source may be used in the compounds of the invention; for example, collagen may be extracted and purified from human or other mammalian source, such as bovine or porcine corium and human placenta, or may be recombinantly or otherwise produced.
• the preparation of purified, substantially non-antigenic collagen in solution from bovine skin is well known in the art. For example, U.S. Patent No. 5,428,022 to Palefsky et al.
• telopeptide-containing collagen Either atelopeptide or telopeptide- containing collagen may be used; however, when collagen from a natural source, such as bovine collagen, is used, atelopeptide collagen is generally preferred, because of its reduced immunogenicity compared to telopeptide- containing collagen.
• Collagen that has not been previously crosslinked by methods such as heat, irradiation, or chemical crosslinking agents is preferred for use in the invention, although previously crosslinked collagen may be used.
• Collagens for use in the present invention are generally, although not necessarily, in aqueous suspension at a concentration between about 20 mg/ml to about 120 mg/ml, preferably between about 30 mg/ml to about 90 mg/ml.
• denatured collagen commonly known as gelatin
• Gelatin may have the added benefit of being degradable faster than collagen.
• Nonfibrillar collagen is generally preferred for use in compounds of the invention, although fibrillar collagens may also be used. The term
• nonfibrillar collagen refers to any modified or unmodified collagen material that is in substantially nonfibrillar form, i.e., molecular collagen that is not tightly associated with other collagen molecules so as to form fibers.
• a solution of nonfibrillar collagen is more transparent than is a solution of fibrillar collagen.
• Collagen types that are nonfibrillar (or microfibrillar) in native form include types IV, VI, and VII.
• Chemically modified collagens that are in nonfibrillar form at neutral pH include succinylated collagen and methylated collagen, both of which can be prepared according to the methods described in U.S. Patent No. 4,164,559 to Miyata et al.
• Methylated collagen which contains reactive amine groups, is a preferred nucleophile-containing component in the compositions of the present invention.
• methylated collagen is a component that is present in addition to first and second components in the matrix-forming reaction of the present invention.
• Methylated collagen is described in, for example, in U.S. Patent No. 5,614,587 to Rhee et al.
• Collagens for use in the compositions of the present invention may start out in fibrillar form, then can be rendered nonfibrillar by the addition of one or more fiber disassembly agent.
• the fiber disassembly agent must be present in an amount sufficient to render the collagen substantially nonfibrillar at pH 7, as described above.
• Fiber disassembly agents for use in the present invention include, without limitation, various biocompatible alcohols, amino acids, inorganic salts, and carbohydrates, with biocompatible alcohols being particularly preferred.
• Preferred biocompatible alcohols include glycerol and propylene glycol.
• Non-biocompatible alcohols such as ethanol, methanol, and isopropanol, are not preferred for use in the present invention, due to their potentially deleterious effects on the body of the patient receiving them.
• Preferred amino acids include arginine.
• Preferred inorganic salts include sodium chloride and potassium chloride.
• carbohydrates such as various sugars including sucrose
• they are not as preferred as other types of fiber disassembly agents because they can have cytotoxic effects in vivo.
• Fibrillar collagen is less preferred for use in the compounds of the invention.
• fibrillar collagen or mixtures of nonfibrillar and fibrillar collagen, may be preferred for use in compounds intended for long-term persistence in vivo.
• the core of the self-reactive compound may also comprise a hydrophobic polymer, including low molecular weight polyfunctional species, although for most uses hydrophilic polymers are preferred.
• hydrophobic polymers herein contain a relatively small proportion of oxygen and/or nitrogen atoms.
• Preferred hydrophobic polymers for use in the invention generally have a carbon chain that is no longer than about 14 carbons. Polymers having carbon chains substantially longer than 14 carbons generally have very poor solubility in aqueous solutions and, as such, have very long reaction times when mixed with aqueous solutions of synthetic polymers containing, for example, multiple nucleophilic groups. Thus, use of short-chain oligomers can avoid solubility-related problems during reaction.
• amphiphilic polymers have a hydrophilic portion and a hydrophobic (or lipophilic) portion.
• the hydrophilic portion can be at one end of the core and the hydrophobic portion at the opposite end, or the hydrophilic and hydrophobic portions may be distributed randomly (random copolymer) or in the form of sequences or grafts (block copolymer) to form the amphiphilic polymer core of the self-reactive compound.
• the hydrophilic and hydrophobic portions may include any of the aforementioned hydrophilic and hydrophobic polymers.
• amphiphilic polymer core can be a hydrophilic polymer that has been modified with hydrophobic moieties (e.g., alkylated PEG or a hydrophilic polymer modified with one or more fatty chains ), or a hydrophobic polymer that has been modified with hydrophilic moieties (e.g., "PEGylated” phospholipids such as polyethylene glycolated phospholipids).
• hydrophobic moieties e.g., alkylated PEG or a hydrophilic polymer modified with one or more fatty chains
• hydrophobic polymer that has been modified with hydrophilic moieties e.g., "PEGylated” phospholipids such as polyethylene glycolated phospholipids.
• the molecular core of the self-reactive compound can also be a low molecular weight compound, defined herein as being a C 2- ⁇ 4 hydrocarbyl or a heteroatom-containing C 2- ⁇ 4 hydrocarbyl, which contains 1 to 2 heteroatoms selected from N, O, S and combinations thereof.
• a molecular core can be substituted with any of the reactive groups described herein.
• Alkanes are suitable C 2 _ ⁇ 4 hydrocarbyl molecular cores.
• alkanes for substituted with a nucleophilic primary amino group and a Y electrophilic group, include, ethyleneamine (H 2 N-CH 2 CH 2 -Y), tetramethyleneamine (H 2 N-(CH 4 )-Y), pentamethyleneamine (H 2 N-(CH 5 )-Y), and hexamethyleneamine (H 2 N-(CH 6 )-Y).
• Low molecular weight diols and polyols are also suitable C 2- ⁇ 4 hydrocarbyls and include trimethylolpropane, di(trimethylol propane), pentaerythritol, and diglycerol.
• Polyacids are also suitable C 2-1 hydrocarbyls, and include trimethylolpropane-based tricarboxylic acid, di(trimethylol propane)- based tetracarboxylic acid, heptanedioic acid, octanedioic acid (suberic acid), and hexadecanedioic acid (thapsic acid).
• Low molecular weight di- and poly-electrophiles are suitable heteroatom-containing C 2- *i 4 hydrocarbyl molecular cores.
• the self-reactive compound of the invention comprises a low-molecular weight material core, with a plurality of acrylate moieties and a plurality of thiol groups.
• the self-reactive compounds are readily synthesized to contain a hydrophilic, hydrophobic or amphiphilic polymer core or a low molecular weight core, functionalized with the desired functional groups, i.e., nucleophilic and electrophilic groups, which enable crosslinking.
• a self-reactive compound having a polyethylene glycol (PEG) core is discussed below.
• PEG polyethylene glycol
• Multi-functionalized forms of PEG are commercially available, and are also easily prepared using known methods. For example, see Chapter 22 of Poly(ethylene Glycol) Chemistry: Biotechnical and Biomedical Applications, J. Milton Harris, ed., Plenum Press, NY (1992).
• Multi-functionalized forms of PEG are of particular interest and include, PEG succinimidyl glutarate, PEG succinimidyl propionate, succinimidyl butylate, PEG succinimidyl acetate, PEG succinimidyl succinamide, PEG succinimidyl carbonate, PEG propionaldehyde, PEG glycidyl ether, PEG- isocyanate, and PEG-vinylsulfone.
• Multi-amino PEGs useful in the present invention include the Jeffamine diamines ("D” series) and triamines ("T” series), which contain two and three primary amino groups per molecule.
• Analogous poly(sulfhydryl) PEGs are also available from Nektar Therapeutics, e.g., in the form of pentaerythritol poly( ethylene glycol) ether tetra-sulfhydryl (molecular weight 10,000). These multi-functionalized forms of PEG can then be modified to include the other desired reactive groups.
• Reaction with succinimidyl groups to convert terminal hydroxyl groups to reactive esters is one technique for preparing a core with electrophilic groups.
• This core can then be modified include nucleophilic groups such as primary amines, thiols, and hydroxyl groups.
• Other agents to convert hydroxyl groups include carbonyldiimidazole and sulfonyl chloride.
• electrophilic groups may be advantageously employed for reaction with corresponding nucleophilic groups.
• Other in situ Crosslinking Materials Numerous other types of in situ forming materials have been described which may be used in combination with an anti-scarring agent in accordance with the invention.
• the in situ forming material may be a biocompatible crosslinked polymer that is formed from water soluble precursors having electrophilic and nucleophilic groups capable of reacting and crosslinking in situ (see, e.g., U.S. Patent No.
• the in situ forming material may be hydrogel that may be formed through a combination of physical and chemical crosslinking processes, where physical crosslinking is mediated by one or more natural or synthetic components that stabilize the hydrogel- forming precursor solution at a deposition site for a period of time sufficient for more resilient chemical crosslinks to form (see, e.g., U.S. Patent No. 6,818,018).
• the in situ forming material may be formed upon exposure to an aqueous fluid from a physiological environment from dry hydrogel precursors (see, e.g., U.S. Patent No. 6,703,047).
• the in situ forming material may be a hydrogel matrix that provides controlled release of relatively low molecular weight therapeutic species by first dispersing or dissolving the therapeutic species within relatively hydrophobic rate modifying agents to form a mixture; the mixture is formed into microparticles that are dispersed within bioabsorbable hydrogels, so as to release the water soluble therapeutic agents in a controlled fashion (see, e.g., 6,632,457).
• the in situ forming material may be a multi-component hydrogel system (see, e.g., U.S. Patent No. 6,379, 373).
• the in situ forming material may be a multi-arm block copolymer that includes a central core molecule, such as a residue of a polyol, and at least three copolymer arms covalently attached to the central core molecule, each copolymer arm comprising an inner hydrophobic polymer segment covalently attached to the central core molecule and an outer hydrophilic polymer segment covalently attached to the hydrophobic polymer segment, wherein the central core molecule and the hydrophobic polymer segment define a hydrophobic core region (see, e.g., 6,730,334).
• a central core molecule such as a residue of a polyol
• each copolymer arm comprising an inner hydrophobic polymer segment covalently attached to the central core molecule and an outer hydrophilic polymer segment covalently attached to the hydrophobic polymer segment, wherein the central core molecule and the hydrophobic polymer segment define a hydrophobic core region (see, e.g., 6,730,334).
• the in situ forming material may include a gel- forming macromer that includes at least four polymeric blocks, at least two of which are hydrophobic and at least one of which is hydrophilic, and including a crosslinkable group (see, e.g., 6,639,014).
• the in situ forming material may be a water-soluble macromer that includes at least one hydrolysable linkage formed from carbonate or dioxanone groups, at least one water-soluble polymeric block, and at least one polymerizable group (see, e.g., U.S. Patent No. 6,177,095).
• the in situ forming material may comprise polyoxyalkylene block copolymers that form weak physical crosslinks to provide gels having a paste-like consistency at physiological temperatures, (see, e.g., U.S. Patent No. 4,911 ,926).
• the in situ forming material may be a thermo-irreversible gel made from polyoxyalkylene polymers and ionic polysaccharides (see, e.g., U.S. Patent No. 5,126,141 ).
• the in situ forming material may be a gel forming composition that includes chitin derivatives (see, e.g., U.S. Patent No. 5,093,319), chitosan-coagulum (see, e.g., U.S. Patent No.
• the in situ forming material may be an in situ modification of alginate (see, e.g., U.S. Patent No. 5,266,326 ).
• the in situ forming material may be formed from ethylenically unsaturated water soluble macromers that can be crosslinked in contact with tissues, cells, and bioactive molecules to form gels (see, e.g., U.S. Patent No. 5,573,934).
• the in situ forming material may include urethane prepolymers used in combination with an unsaturated cyano compound containing a cyano group attached to a carbon atom, such as cyano(meth)acrylic acids and esters thereof (see, e.g., U.S. Patent No. 4,740,534).
• the in situ forming material may be a biodegradable hydrogel that polymerizes by a photoinitiated free radical polymerization from water soluble macromers (see, e.g., U.S. Patent No.
• the in situ forming material may be formed from a two component mixture including a first part comprising a serum albumin protein in an aqueous buffer having a pH in a range of about 8.0-11.0, and a second part comprising a water-compatible or water-soluble bifunctional crosslinking agent, (see, e.g., U.S. Patent No. 5,583,114).
• in situ forming materials that can be used include those based on the crosslinking of proteins.
• the in situ forming material may be a biodegradable hydrogel composed of a recombinant or natural human serum albumin and poly(ethylene) glycol polymer solution whereby upon mixing the solution cross-links to form a mechanical non-liquid covering structure which acts as a sealant.
• the in situ forming material may be composed of two separate mixtures based on fibrinogen and thrombin which are dispensed together to form a biological adhesive when intermixed either prior to or on the application site to form a fibrin sealant. See e.g., U.S. Patent No. 6,764,467.
• the in situ forming material may be composed of ultrasonically treated collagen and albumin which form a viscous material that develops adhesive properties when crosslinked chemically with glutaraldehyde and amino acids or peptides. See e.g., U.S. Patent No. 6,310,036.
• the in situ forming material may be a hydrated adhesive gel composed of an aqueous solution consisting essentially of a protein having amino groups at the side chains (e.g., gelatin, albumin) which is crosslinked with an N-hydroxyimidoester compound. See e.g., U.S. Patent No. 4,839,345.
• the in situ forming material may be a hydrogel prepared from a protein or polysaccharide backbone (e.g., albumin or polymannuronic acid) bonded to a cross-linking agent (e.g., polyvalent derivatives of polyethylene or polyalkylene glycol). See e.g., U.S. Patent No. 5,514,379.
• the in situ forming material may be composed of a polymerizable collagen composition that is applied to the tissue and then exposed to an initiator to polymerize the collagen to form a seal over a wound opening in the tissue. See e.g., U.S. Patent No. 5,874,537.
• the in situ forming material may be a two component mixture composed of a protein (e.g., serum albumin) in an aqueous buffer having a pH in the range of about 8.0-11.0 and a water-soluble bifunctional polyethylene oxide type crosslinking agent, which transforms from a liquid to a strong, flexible bonding composition to seal tissue in situ.
• a protein e.g., serum albumin
• a water-soluble bifunctional polyethylene oxide type crosslinking agent which transforms from a liquid to a strong, flexible bonding composition to seal tissue in situ.
• the in situ forming material may be composed of a protein, a surfactant, and a lipid in a liquid carrier, which is crosslinked by adding a crosslinker and used as a sealant or bonding agent in situ. See e.g., U.S.
• the in situ forming material may be composed of two enzyme-free liquid components that are mixed by dispensing the components into a catheter tube deployed at the vascular puncture site, wherein, upon mixing, the two liquid components chemically cross-link to form a mechanical non-liquid matrix that seals a vascular puncture site. See e.g., U.S. Patent Application Nos. 2002/0161399A1 and 2001/0018598A1.
• the in situ forming material may be a cross-linked albumin composition composed of an albumin preparation and a carbodiimide preparation which are mixed under conditions that permit crosslinking of the albumin for use as a bioadhesive or sealant. See e.g., PCT Publication No. WO 99/66964.
• the in situ forming material may be composed of collagen and a peroxidase and hydrogen peroxide, such that the collagen is crosslinked to from a semi-solid gel that seals a wound. See e.g., PCT Publication No. WO 01/35882.
• in situ forming materials that can be used include those based on isocyanate or isothiocyanate capped polymers.
• the in situ forming material may be composed of isocyanate-capped polymers that are liquid compositions which form into a solid adhesive coating by in situ polymerization and crosslinking upon contact with body fluid or tissue.
• the in situ forming material may be a moisture-curing sealant composition composed of an active isocyanato-terminated isocyanate prepolymer containing a polyol component with a molecular weight of 2,000 to 20,000 and an isocyanurating catalyst agent. See e.g., U.S. Patent No. 5,206,331.
• the reagents can undergo an electrophilic-nucleophilic reaction to produce a crosslinked matrix.
• Polymers containing and/or terminated with nucleophilic groups such as amine, sulfhydryl, hydroxyl, -PH 2 or CO-NH-NH 2 can be used as the nucleophilic reagents and polymers containing and/or terminated with electrophilic groups such as succinimidyl, carboxylic acid, aldehyde, epoxide, isocyanate, vinyl, vinyl sulfone, maleimide, -S-S-(C 5 H 4 N) or activated esters, such as are used in peptide synthesis can be used as the electrophilic reagents.
• nucleophilic groups such as amine, sulfhydryl, hydroxyl, -PH 2 or CO-NH-NH 2
• electrophilic groups such as succinimidyl, carboxylic acid, aldehyde, epoxide, isocyanate, vinyl, vinyl sulfone, maleimide, -S-S-(C 5 H 4 N) or
• a 4- armed thiol derivatized poly(ethylene glycol) e.g., pentaerythritol poly(ethyIene glycol)ether tetra-sulfhydryl
• a 4 armed NHS-derivatized polyethylene glycol e.g., pentaerythritol poly(ethylene glycol)ether tetra- succinimidyl glutarate
• pH > about 8 Representative examples of compositions that undergo such electrophilic-nucleophilic crosslinking reactions are described, for example, in U.S. Patent. Nos.
• the electrophilic- or nucleophilic- terminated polymers can further comprise a polymer that can enhance the mechanical and/or adhesive properties of the in situ forming compositions.
• This polymer can be a degradable or non-degradable polymer.
• the polymer may be collagen or a collagen derivative, for example methylated collagen.
• An example of an in situ forming composition uses pentaerythritol poly(ethylene g!ycol)ether tetra-sulfhydryl) (4-armed thiol PEG), pentaerythritol poly(ethylene glycol)ether tetra-succinimidyl glutarate) (4-armed NHS PEG) and methylated collagen as the reactive reagents.
• This composition when mixed with the appropriate buffers can produce a crosslinked hydrogel.
• the reagents that can form a covalent bond with the tissue to which it is applied may be used.
• Polymers containing and/or terminated with electrophilic groups such as succinimidyl, aldehyde, epoxide, isocyanate, vinyl, vinyl sulfone, maleimide, -S-S-(C 5 H 4 N) or activated esters, such as are used in peptide synthesis may be used as the reagents.
• a 4 armed NHS-derivatized polyethylene glycol e.g., pentaerythritol poly(ethylene glycol)ether tetra-succinimidyl glutarate
• the 4 armed NHS-derivatized polyethylene glycol is applied to the tissue under basic conditions (pH > about 8).
• basic conditions pH > about 8
• compositions of this nature that may be used are disclosed in PCT Application Publication No. WO 04/060405 and WO 04/060346, and U.S. Patent Application No. 10/749,123.
• the in situ forming material polymer can be a polyester.
• Polyesters that can be used in in situ forming compositions include poly(hydroxyesters).
• the polyester can comprise the residues of one or more of the monomers selected from lactide, lactic acid, glycolide, glycolic acid, e-caprolactone, gamma-caprolactone, hydroxyvaleric acid, hydroxybutyric acid, beta-butyrolactone, gamma- butyrolactone, gamma-valerolactone, ⁇ -decanolactone, ⁇ -decanolactone, trimethylene carbonate, 1 ,4-dioxane-2-one or 1 ,5-dioxepan-2one. Representative examples of these types of compositions are described in U.S. Patent. Nos.
• the electrophilic-terminated polymer can be partially or completely replaced by a small molecule or oligomer that comprises an electrophilic group (e.g., disuccinimidyl glutarate).
• the nucleophilic-terminated polymer can be partially or completely replaced by a small molecule or oligomer that comprises a nucleophilic group (e.g., dicysteine, dilysine, trilysine, etc.).
• in situ forming materials that can be used include those based on the crosslinking of proteins (described in, for example, U.S.
• Other examples of in situ forming materials can include reagents that comprise one or more cyanoacrylate groups.
• reagents can be used to prepare a poly(alkylcyanoacrylate) or poly(carboxyalkylcyanoacrylate) (e.g., poly(ethylcyanoacrylate), poly(butylcyanoacrylate), poly(isobutylcyanoacrylate), poly(hexylcyanoacrylate), poly(methoxypropylcyanoacrylate), and poly(octylcyanoacrylate)).
• poly(alkylcyanoacrylate) or poly(carboxyalkylcyanoacrylate) e.g., poly(ethylcyanoacrylate), poly(butylcyanoacrylate), poly(isobutylcyanoacrylate), poly(hexylcyanoacrylate), poly(methoxypropylcyanoacrylate), and poly(octylcyanoacrylate)
• cyanoacrylates examples include DERMABOND, INDERMIL, GLUSTITCH, VETBOND, HISTOACRYL, TISSUMEND, HISTOACRYL BLUE and ORABASE SOOTHE-N-SEAL LIQUID PROTECTANT.
• the cyanoacrylate compositions may further comprise additives to stabilize the reagents and/or alter the rate of reaction of the cyanoacrylate, and/or plasticize the poly( cyanoacrylate), and/or alter the rate of degradation of the poly( cyanoacrylate).
• a trimethylene carbonate based polymer or an oxalate polymer of poly( ethylene glycol) or a ⁇ -caprolactone based copolymer may be mixed with a 2- alkoxyalkylcyanoacrylate (e.g., 2-methoxypropylcyanoacrylate).
• a 2- alkoxyalkylcyanoacrylate e.g., 2-methoxypropylcyanoacrylate
• Representative examples of these compositions are described in U.S. Patent Nos. 5,350,798 and 6,299,631.
• the cyanoacrylate composition can be prepared by capping heterochain polymers with a cyanoacrylate group.
• the cyanoacrylate-capped heterochain polymer preferably has at least two cyanoacrylate ester groups per chain.
• the heterochain polymer can comprise an absorbable poly(ester), poly(ester-carbonate), poly(ether-carbonate) and poly(ether-ester).
• the poly(ether-ester)s described in U.S. Patent Nos. 5,653,992 and 5,714,159 can also be used as the heterochain polymers.
• a triaxial poly( ⁇ -caprolactone-co-trimethylene carbonate) is an example of a poly(ester-carbonate) that can be used.
• the heterochain polymer may be a polyether.
• polyethers examples include poly(ethylene glycol), poly(propylene glycol) and block copolymers of poly( ethylene glycol) and poly(propylene glycol) (e.g., PLURONICS group of polymers including but not limited to PLURONIC F127 or F68). Representative examples of these compositions are described in U.S. Patent No. 6,699,940.
• the biologically active ant- infective and/or fibrosis-inhibiting agent can be delivered with a non-polymeric compound (e.g., a carrier).
• non-polymeric carriers can include sucrose derivatives (e.g., sucrose acetate isobutyrate, sucrose oleate), sterols such as cholesterol, stigmasterol, ⁇ -sitosterol, and estradiol; cholesteryl esters such as cholesteryl stearate; C ⁇ 2 -C 24 fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, and lignoceric acid; Cis -C 36 mono-, di- and triacylglycerides such as glyceryl monooleate, glyceryl monolinoleate, glyceryl monolaurate, glyceryl monodocosanoate, glyceryl monomyristate, glyceryl monodicenoate, glyceryl dipalmitate, glyceryl didocosanoate, glyceryl dimyristate, g
• the therapeutic compositions include (i) a fibrosis-inhibiting agent and/or (ii) an anti-infective agent.
• the therapeutic compositions may include one or more additional therapeutic agents (such as described above), for example, anti- inflammatory agents, anti-thrombotic agents, and/ or anti-platelet agents.
• surfactants e.g., PLURONICS, such as F-127, L-122, L-101 , L-92, L-81 , and L-61
• preservatives e.g., anti-oxidants.
• the present invention provides compositions comprising i) an anti-fibrotic agent and ii) a polymer or a compound that forms a polymer in situ.
• compositions comprising i) an anti-fibrotic agent and ii) a polymer or a compound that forms a polymer in situ.
• 3a An anti-fibrotic agent that inhibits fibroblast migration.
• 4a An anti-fibrotic agent that inhibits fibroblast proliferation.
• An anti-fibrotic agent inhibits tissue remodeling.
• 9a An anti-fibrotic agent that is a chemokine receptor antagonist.
• 10a An anti-fibrotic agent that is a cell cycle inhibitor.
• An anti-fibrotic agent that is a taxane.
• An anti-fibrotic agent that is an anti-microtubule agent.
• An anti-fibrotic agent that is a cathepsin inhibitor 14a.
• An anti-fibrotic agent that is an analogue or derivative of paclitaxel 15a.
• An anti-fibrotic agent that is a vinca alkaloid.
• 17a An anti-fibrotic agent that is camptothecin or an analogue or derivative thereof.
• 18a An anti-fibrotic agent that is a podophyllotoxin.
• An anti-fibrotic agent that is an anthracycline.
• An anti-fibrotic agent that is doxorubicin or an analogue or derivative thereof.
• 22a An anti-fibrotic agent that mitoxantrone or an analogue or derivative thereof.
• 23a An anti-fibrotic agent that is a platinum compound.
• An anti-fibrotic agent that is a nitrosourea.
• An anti-fibrotic agent that is a nitroimidazole.
• 26a An anti-fibrotic agent that is a folic acid antagonist.
• 27a An anti-fibrotic agent that is a cytidine analogue.
• An anti-fibrotic agent that is a pyrimidine analogue.
• An anti-fibrotic agent that is a purine analogue.
• An anti-fibrotic agent that is a nitrogen mustard or an analogue or derivative thereof.
• An anti-fibrotic agent that is a mytomicin or an analogue or derivative thereof.
• An anti-fibrotic agent that is an alkyl sulfonate 35a.
• An anti-fibrotic agent that is a nicotinamide or an analogue or derivative thereof.
• An anti-fibrotic agent that is a halogenated sugar or an analogue or derivative thereof.
• An anti-fibrotic agent that is a topoisomerase inhibitor.
• An anti-fibrotic agent that is a DNA cleaving agent.
• An anti-fibrotic agent inhibits adenosine deaminase.
• An anti-fibrotic agent inhibits purine ring synthesis.
• An anti-fibrotic agent that is a nucleotide interconversion inhibitor.
• An anti-fibrotic agent inhibits dihydrofolate reduction. 47a. An anti-fibrotic agent blocks thymidine monophosphate.
• An anti-fibrotic agent causes DNA damage.
• An anti-fibrotic agent that is a DNA intercalation agent.
• An anti-fibrotic agent that is a RNA synthesis inhibitor.
• 51a An anti-fibrotic agent that is a pyrimidine synthesis inhibitor.
• 52a An anti-fibrotic agent that inhibits ribonucleotide synthesis or function.
• 54a An anti-fibrotic agent that inhibits DNA synthesis.
• 55a An anti-fibrotic agent that causes DNA adduct formation.
• An anti-fibrotic agent that is a cyclin dependent protein kinase inhibitor.
• An anti-fibrotic agent that is an epidermal growth factor kinase inhibitor.
• An anti-fibrotic agent that is an elastase inhibitor.
• An anti-fibrotic agent that is a factor Xa inhibitor.
• An anti-fibrotic agent that is a farnesyltransferase inhibitor.
• 63a An anti-fibrotic agent that is a fibrinogen antagonist.
• 64a An anti-fibrotic agent that is a guanylate cyclase stimulant.
• An anti-fibrotic agent that is a heat shock protein 90 antagonist.
• 66a An anti-fibrotic agent that is geldanamycin or an analogue or derivative thereof.
• 67a An anti-fibrotic agent that is a guanylate cyclase stimulant.
• An anti-fibrotic agent that is a HMGCoA reductase inhibitor.
• An anti-fibrotic agent that is simvastatin or an analogue or derivative thereof.
• An anti-fibrotic agent that is a hydroorotate dehydrogenase inhibitor.
• An anti-fibrotic agent that is an IKK2 inhibitor.
• An anti-fibrotic agent that is an IRAK antagonist.
• An anti-fibrotic agent that is an IL-4 agonist.
• 76a An anti-fibrotic agent that is an immunomodulatory agent.
• 77a An anti-fibrotic agent that is sirolimus or an analogue or derivative thereof.
• An anti-fibrotic agent that is a nitric oxide inhibitor.
• 79a An anti-fibrotic agent that is everolimus or an analogue or derivative thereof.
• 80a An anti-fibrotic agent that is tacrolimus or an analogue or derivative thereof.
• An anti-fibrotic agent that is a TNF alpha inhibitor.
• An anti-fibrotic agent that is auranofin or an analogue or derivative thereof.
• An anti-fibrotic agent that is 27-0-demethylrapamycin or an analogue or derivative thereof.
• 86a An anti-fibrotic agent that is gusperimus or an analogue or derivative thereof.
• 87a An anti-fibrotic agent that is pimecrolimus or an analogue or derivative thereof.
• IMPDH inosine monophosphate dehydrogenase
• An anti-fibrotic agent that is mycophenolic acid or an analogue or derivative thereof.
• An anti-fibrotic agent that is 1-alpha-25 dihydroxy vitamin D 3 or an analogue or derivative thereof.
• An anti-fibrotic agent that is a leukotriene inhibitor.
• An anti-fibrotic agent that is a MCP-1 antagonist.
• An anti-fibrotic agent that is a MMP inhibitor.
• An anti-fibrotic agent that is a p38 MAP kinase inhibitor.
• An anti-fibrotic agent that is a p38 MAP kinase inhibitor, wherein the p38 MAP kinase inhibitor is SB 202190.
• An anti-fibrotic agent that is a phosphodiesterase inhibitor.
• 101a An anti-fibrotic agent that is a TGF beta inhibitor.
• 102a An anti-fibrotic agent that is a thromboxane A2 antagonist.
• An anti-fibrotic agent that is a TNF alpha antagonist.
• An anti-fibrotic agent that is a TACE inhibitor.
• An anti-fibrotic agent that is a tyrosine kinase inhibitor.
• An anti-fibrotic agent that is a vitronectin inhibitor.
• An anti-fibrotic agent that is a fibroblast growth factor inhibitor.
• An anti-fibrotic agent that is a protein kinase inhibitor.
• An anti-fibrotic agent that is a PDGF receptor kinase inhibitor.
• An anti-fibrotic agent that is an endothelial growth factor receptor kinase inhibitor.
• 111a An anti-fibrotic agent that is a retinoic acid receptor antagonist.
• 112a An anti-fibrotic agent that is a platelet derived growth factor receptor kinase inhibitor.
• An anti-fibrotic agent that is a fibrinogen antagonist.
• An anti-fibrotic agent that is an antimycotic agent.
• An anti-fibrotic agent that is an antimycotic agent, wherein the antimycotic agent that is sulconizole.
• An anti-fibrotic agent that is a bisphosphonate.
• 117a An anti-fibrotic agent that is a phospholipase A1 inhibitor.
• 118a An anti-fibrotic agent that is a histamine H1/H2/H3 receptor antagonist.
• An anti-fibrotic agent that is a macrolide antibiotic.
• An anti-fibrotic agent that is a GPIIb/llla receptor antagonist.
• An anti-fibrotic agent that is an endothelin receptor antagonist.
• An anti-fibrotic agent that is a peroxisome proliferator- activated receptor agonist 122a.
• An anti-fibrotic agent that is an estrogen receptor agent 122a.
• An anti-fibrotic agent that is a somastostatin analogue.
• An anti-fibrotic agent that is a neurokinin 1 antagonist.
• An anti-fibrotic agent that is a neurokinin 3 antagonist.
• An anti-fibrotic agent that is a DNA topoisomerase ATP hydrolyzing inhibitor.
• An anti-fibrotic agent that is an angiotensin I converting enzyme inhibitor 130a.
• An anti-fibrotic agent that is an angiotensin II antagonist 130a.
• An anti-fibrotic agent that is an enkephalinase inhibitor 133a.
• An anti-fibrotic agent that is a protein kinase C inhibitor.
• An anti-fibrotic agent that is a ROCK (rho-associated kinase) inhibitor.
• An anti-fibrotic agent that is a CXCR3 inhibitor.
• An anti-fibrotic agent that is an Itk inhibitor.
• 139a An anti-fibrotic agent that is a PPAR agonist.
• An anti-fibrotic agent that is an immunosuppressant that is an immunosuppressant.
• An anti-fibrotic agent that is an Erb inhibitor.
• An anti-fibrotic agent that is an apoptosis agonist.
• An anti-fibrotic agent that is a lipocortin agonist e.g., a lipocortin agonist.
• An anti-fibrotic agent that is a VCAM-1 antagonist e.g., a VCAM-1 antagonist.
• An anti-fibrotic agent that is a collagen antagonist.
• compositions comprising each of the foregoing 146 (i.e., 1a through 145a) listed anti-fibrotic agents or classes of anti-fibrotic agents, with each of the following 98 (i.e., 1 b through 97b) polymers and compounds:
• 2b A polymer that reacts with mammalian tissue.
• b A polymer that is a naturally occurring polymer.
• b A polymer that is a protein.
• b A polymer that is a carbohydrate.
• b A polymer that is biodegradable.
• b A polymer that is crosslinked and biodegradable.
• b A polymer that nonbiodegradable.
• 27b A synthetic polymer.
• 28b A polymer formed from reactants comprising a synthetic isocyanate-containing compound.
• a synthetic compound containing at least two thiol groups is a synthetic compound containing at least two thiol groups.
• a synthetic compound containing at least three thiol groups is a synthetic compound containing at least three thiol groups.
• 36b A polymer formed from reactants comprising a synthetic compound containing at least four thiol groups.
• 37b A synthetic compound containing at least four thiol groups.
• a polymer formed from reactants comprising a synthetic amino-containing compound comprising a synthetic amino-containing compound.
• 39b A synthetic amino-containing compound.
• 40b A polymer formed from reactants comprising a synthetic compound containing at least two amino groups.
• 41 b A synthetic compound containing at least two amino groups.
• 42b A polymer formed from reactants comprising a synthetic compound containing at least three amino groups.
• a synthetic compound comprising at least two carbonyl- oxygen-succinimidyl groups.
• 51 b A synthetic compound comprising at least three carbonyl- oxygen-succinimidyl groups.
• 52b A polymer formed from reactants comprising a synthetic compound comprising at least four carbonyl-oxygen-succinimidyl groups.
• 53b A synthetic compound comprising at least four carbonyl- oxygen-succinimidyl groups.
• a synthetic compound comprising both polyalkylene oxide and biodegradable polyester blocks.
• a polymer formed from reactants comprising a synthetic polyalkylene oxide-containing compound having reactive amino groups.
• 59b A synthetic polyalkylene oxide-containing compound having reactive amino groups.
• 60b A polymer formed from reactants comprising a synthetic polyalkylene oxide-containing compound having reactive thiol groups.
• a synthetic polyalkylene oxide-containing compound having reactive thiol groups is provided.
• a polymer formed from reactants comprising a synthetic polyalkylene oxide-containing compound having reactive carbonyl-oxygen- succinimidyl groups.
• a synthetic polyalkylene oxide-containing compound having reactive carbonyl-oxygen-succinimidyl groups having reactive carbonyl-oxygen-succinimidyl groups.
• 64b A polymer formed from reactants comprising a synthetic compound comprising a biodegradable polyester block.
• 65b A synthetic compound comprising a biodegradable polyester block.
• a polymer formed from reactants comprising a synthetic polymer formed in whole or part from lactic acid or lactide.
• 67b A synthetic polymer formed in whole or part from lactic acid or lactide.
• a polymer formed from reactants comprising a synthetic polymer formed in whole or part from glycolic acid or glycolide.

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