WO2008067288A2 - Procédés permettant d'inhiber la réaction à corps étranger face à des matériaux implantés - Google Patents

Procédés permettant d'inhiber la réaction à corps étranger face à des matériaux implantés Download PDF

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WO2008067288A2
WO2008067288A2 PCT/US2007/085598 US2007085598W WO2008067288A2 WO 2008067288 A2 WO2008067288 A2 WO 2008067288A2 US 2007085598 W US2007085598 W US 2007085598W WO 2008067288 A2 WO2008067288 A2 WO 2008067288A2
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rho
inhibitors
cells
group
gtpase
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WO2008067288A3 (fr
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Themis R. Kyriades
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Yale University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • 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
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • 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
    • A61L29/00—Materials for catheters, medical tubing, cannulae, or endoscopes or for coating catheters
    • A61L29/14—Materials characterised by their function or physical properties, e.g. lubricating compositions
    • A61L29/16—Biologically active materials, e.g. therapeutic substances
    • 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
    • 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
    • A61L2300/432—Inhibitors, antagonists
    • A61L2300/434—Inhibitors, antagonists of enzymes

Definitions

  • aspects of the disclosure are generally related to methods for inhibiting the formation of multinucleated foreign body giant cells.
  • the disclosed methods may be used to inhibit damage to implanted foreign body materials and to enhance their biocompatibility.
  • FBR foreign body reaction
  • the FBR has been implicated in the malfunction and failure of numerous devices and implants including glucose sensors, cochlear implants, breast augmentation prosthesis and artificial joints (Tang and Eaton, Am. J Clin. Pathol, 103:466-471 (1995); Woodward, Diabetes Care, 5 :278-281 (1982); Ratner, J Biomed. Mater.
  • FBGC foreign body giant cells
  • FBGC are believed to be key mediators of the inflammatory response to implanted materials.
  • FBGC can create high concentrations of enzymes that can cause extensive surface damage (Zhao, et al , J. Biomed. Mater. Res., 25:177-183 (1991)).
  • FBGC can generate particulate debris that can contribute to the persistence of the FBR.
  • Foreign body giant cells are also thought to be a source of chemokines such as IL-8 which recruits neutrophils and lymphocytes to the site.
  • chemokines such as IL-8 which recruits neutrophils and lymphocytes to the site.
  • Biomaterial-induced FBGC formation and the associated fibrous encapsulation result in a physical walling off effect that enhances material degradation and prevents appropriate molecular transport and vascularization (Anderson, Annual Review of Materials Research, 31:81-110 (2001); Zhao, et al., J. Biomed Mater. Res., 25:177-183 (1991); Kyriakides and Bornstein, Thromb. Haemost. , 90:986-992 (2003)).
  • the methods include inhibiting the formation of foreign body giant cells by administering an effective amount of an antagonist of Rho GTPase activity.
  • the methods include contacting an implanted material or the adjacent tissue or bodily fluids with an effective amount of a Rho GTPase antagonist to prevent formation of multinucleated foreign body giant cells at the implantation site.
  • Inhibitors of Rho GTPase activity may be provided separately from the implanted material, or alternatively, may be provided as a component of the external surface of the implanted material.
  • the methods include contacting monocytes or macrophages with inhibitors of Rho family GTPase activity in amounts effective to inhibit fusion of these cells to form multinucleated foreign body giant cells.
  • Inhibitors of Rho GTPase activity are preferably inhibitors of Rac family GTPase activity.
  • the small molecule Rac inhibitor NSC23766 is used.
  • the described methods may be used to protect implanted materials from surface damage and encapsulation, thereby enhancing the biocompatibility of an implanted material. Examples demonstrate that the methods are effective to inhibit foreign body giant cell formation in response to implanted foreign body materials.
  • Figure IA is a graph showing percent fusion of murine monocytes from control (C57BL6) and MCP-I knock-out (KO) mice induced to fuse into multinucleated giant cells through concurrent addition of GM-CSF and IL-4.
  • Figure IB is a graph showing average number of nuclei per foreign body giant cell (FBGC) formed from murine monocytes from control (C57BL6) and MCP-I knock-out (KO) mice induced to fuse through concurrent addition of GM-CSF and IL-4.
  • Figure 2 A is a graph showing the average number of FBGCs per high-powered field (HPF) formed from the fusion of murine monocytes in the absence or presence of two different concentrations of the Rac GTPase inhibitor, NSC23766.
  • HPF high-powered field
  • Figure 2B is a graph showing the average number of nuclei per FBGC formed from the fusion of murine monocytes in the absence or presence of two different concentrations of the Rac GTPase inhibitor, NSC23766.
  • Figure 3 is a graph showing the average number of nuclei per FBGC formed from the fusion of murine monocytes in the absence or presence of three different concentrations of the Rho kinase inhibitor, Y-27632.
  • Figure 4 is a graph showing the average number of FBGCs per 200 ⁇ M of histological section taken from the implantation site of mice implanted with EVAc scaffolds with or without NSC23766.
  • Formal body giant cells refers to multinucleated cells which are derived from the fusion of monocytes or macrophages which are recruited to a site of implantation.
  • Ras or Ras superfamily proteins encompass a large family of GTP binding/GTP hydrolyzing monomeric proteins. Ras family includes the Ras, Rho, Rab, Arf, and Ran subfamilies of GTPases.
  • Rho GTPases or “Rho family GTPases” refer to a subfamily of Ras superfamily and are small, membrane-bound, Ras-related GTP-binding proteins that function by binding and hydrolyzing GTP.
  • Rho GTPases function as molecular switches, cycling between an inactive GDP- bound conformation and an active GTP-bound conformation and include RhoA, RhoB, RhoC, Cdc42, Racl, Rac2, Rac3, TClO, RhoG, RhoD, Chp, WRCHl 5 TCL, and RIF.
  • Rac GTPase or “Rac protein or polypeptide” refer to Racl, Rac2, and/or Rac3.
  • a "farnesyl protein transferase inhibitor” or “FPT inhibitor” or “FPTase inhibitor” or “FTI” is defined herein as a compound which: (i) potently inhibits FPTase (but generally not geranylgeranyl protein transferase I) and (ii) blocks intracellular farnesylation of Ras. FPTase catalyzes the addition of an isoprenyl lipid moiety onto a cysteine residue present near the carboxy-terminus of the Ras protein. This is the first step in a post-translational processing pathway that is essential for both Ras membrane-association and Ras-induced oncogenic transformation.
  • I Compositions for inhibiting foreign body giant cell formation
  • Rho GTPases constitute one branch of the Ras superfamily of GTPases. Rho proteins share approximately 30 percent amino acid identity with the Ras proteins. Rho GTPases are small membrane-bound GTP-binding proteins that function by binding and hydrolyzing GTP. Rho GTPases function as molecular switches, cycling between an inactive GDP-bound conformation and an active GTP-bound conformation. In their active GTP-bound conformation, Rho GTPases can bind to multiple effector proteins and thus affect their function.
  • Rho GTPases include at least 14 identified proteins including RhoA, RhoB, RhoC, Cdc42, Racl, Rac2, Rac3, TClO, RhoG, RhoD, Chp, WRCHl, TCL and RIF.
  • Rho GTPase inhibitors include any agent that modulates Rho GTPase activity or expression in a living body.
  • Rho GTPase inhibitors useful for inhibiting Rho GTPase protein activity include inhibitors of prenyl protein transferases, famesyl protein transferases and geranylgeranyl protein transferases.
  • Other agents useful for the practice of the provided methods include toxins, the small molecule NSC23766, siRNAs, dominant negative mutants, ribozymes, antibodies, blocking peptides and combinations thereof.
  • the Rho GTPase inhibitor can be one or more prenylation inhibitors, such as disclosed by U.S. Patent Nos. 6,649,638, 5,420,245; 5,574,025; 5,523,430; 5,602,098; 5,631,401; 5,705,686; 5,238,922; 5,470,832; and 6,191,147.
  • the Rho GTPase inhibitor includes one or more inhibitors of famesyl protein transferase (FPTase), prenyl-protein transferase or geranylgeranyl-protein transferase, as described in U.S. Patent Nos.
  • Farnesyl transferase inhibitors generally fall into two classes: analogs of farnesyl diphosphate; and protein substrates for farnesyl transferase. Farnesyl transferase inhibitors have been described in U.S. Patent Nos. 5,756,528, 5,141,851, 5,817,678, 5,830,868, 5,834,434, and 5,773,455.
  • L-739,749 (a peptidomimetic analog of the C-A-A-X sequence)
  • L-744 S 832 a peptidomimetic analog of the C-A-A-X sequence
  • SCH 44342 (l-(4- ⁇ yridylacetyi)-4-(8-chloro-5,6 dihydro-IIH benzo[5,6]cyclohepta [1,2- b]pyridin-l l-yhdene)piperid ⁇ ne
  • BZA-5B (a benzodiazepine peptidomimetic)
  • FTI-276 (a C-A-A-X peptidomimetic)
  • B 1086 (a C-A- A-X peptidomimetic).
  • the Rho GTPase inhibitor includes one or more inhibitors of geranylgeranyl-protein transferase (GGT), for example, as have been described in U.S. Patent No. 5,470,832.
  • the Rho GTPase inhibitor includes one or more toxins such as toxins A and B from C. difficile and C. sordellii lethal toxin (LT).
  • Racl and Rac2 can be inhibited when Rho is specifically ADP ribosylated by C3 enzyme, which is one of the botulinum toxins, and Staphylococcal toxin EDIN (Narumiya and Morii, Cell Signal.
  • Rho GTPase inhibitor is the small molecule, NSC23766, as described in U.S. Published Application No. 2006/0004032. While not wishing to be bound by theory, NSC23766 is believed to work to inhibit activation of Rac proteins by inhibiting their association with guanine nucleotide exchange factors (GEFs). Binding of GEF proteins to Rho GTPases normally causes a conformational change in Rho GTPases and thereby causes dissociation of GDP from these proteins and exchange of GDP for GTP.
  • GEFs guanine nucleotide exchange factors
  • Antibodies may also be used to inhibit the activity of Rho GTPases.
  • Methods for preparing monoclonal and polyclonal antibodies are well known to those of skill in the art and are set forth, for example, in chapters five and six of Antibodies A Laboratory Manual, E. Harlow and D. Lane, Cold Spring Harbor Laboratory (1988).
  • Antibody production includes not only the stimulation of an immune response by injection into animals, but also analogous processes such as the production of synthetic antibodies, and the screening of recombinant immunoglobulin libraries for specific-binding molecules (Orlandi, et al. Proc. Natl. Acad. Sci. USA, 86:3833 (1989); Huse, et at, Science, 256:1275 (1989)), as well as the in vitro stimulation of lymphocyte populations.
  • Rho GTPase activity may also be inhibited using blocking peptides.
  • Useful peptides bind specifically to Rho GTPases or binding partners of Rho GTPases. Especially useful peptides are those that inhibit the association of Rho GTPases with effectors of guanine nucleotide binding such as guanine nucleotide exchange factors and GTPase activating proteins.
  • Other useful peptides include those that prevent binding of Rho GTPases to effector-like proteins such as integrins, growth factor receptors, tyrosine kinases, and lipid kinases.
  • Other useful peptides include those that inhibit binding, release, or hydrolysis of the guanine nucleotides GDP or GTP by a Rho GTPase.
  • Rho GTPases that have a "dominant negative" effect on activation of endogenous Rho GTPases are also useful to inhibit Rho GTPase activity.
  • Dominant negative mutants of Rho GTPase proteins are mutants to which GDP remains bound despite the ability of the Rho GTPase to interact with guanine nucleotide exchange factors (GEFs).
  • GEFs guanine nucleotide exchange factors
  • Dominant negative Rho GTPases are thought to inhibit the activation of endogenous wild-type Rho GTPases through the sequestration of GEFs.
  • Dominant negative mutants of many Rho GTPases are well known in the art, and include RhoN19, RacN17, and Cdc42N17.
  • Ribozymes may be used to decrease the expression of Rho GTPases, such as ribozymes which target Rho GTPase mRNA.
  • Ribozymes are catalytic RNA molecules that can cleave nucleic acid molecules having a sequence that is completely or partially homologous to the sequence of the ribozyme. It is possible to design ribozyme transgenes that encode RNA ribozymes that specifically pair with a target RNA and cleave the phosphodiester backbone at a specific location, thereby functionally inactivating the target RNA. In carrying out this cleavage, the ribozyme is not itself altered, and is thus capable of recycling and cleaving other molecules. The inclusion of ribozyme sequences within antisense RNAs confers RNA-cleaving activity upon them, thereby increasing the activity of the antisense constructs.
  • Ribozymes useful to reduce Rho GTPase mRNA expression typically include a hybridizing region, of at least about nine nucleotides, which is complementary in nucleotide sequence to at least part of the target Rho
  • GTPase mRNA and a catalytic region which is adapted to cleave the target Rho GTPase mRNA
  • Rho GTPase mRNA see generally, EPA No. 0 321 201; WO88/04300; Haseloff and Gerlach, Nature, 334:585-591 (1988); Fedor and Uhlenbeck, Proc. Natl Acad. Set USA 87:1668-1672 (1990); Cech and Bass, Ann. Rev. Biochem., 55:599-629 (1986)
  • Specific ribozyme cleavage sites within any potential RNA target can be identified by scanning the target RNA for ribozyme cleavage sites which include, for example, the following sequences: GUA, GUU, and GUC.
  • RNA sequences of between 9 and 20 ribonucleotides corresponding to the region of the target polynucleotide containing the cleavage site can be evaluated for secondary structural features which can render the oligonucleotide inoperable.
  • Antisense molecules and ribozymes useful to decrease Rho GTPase mRNA levels can be prepared by any method known in the art for the synthesis of nucleic acid molecules. Double-stranded nucleic acid molecules may also be used to reduce expression levels of Rho GTPases and thereby inhibit their function.
  • Double-stranded RNA (dsRNA) containing complementary sequences to portions of Rho GTPase mRNAs may be used to induce RNA interference (RNAi) of a targeted mRNA.
  • RNAi is thought to work by binding of the target mRNA by the complementary dsRNA which targets the mRNA for degradation by an enzymatic process.
  • dsRNA molecules of between 20 and 25 nucleotides in length are typically used to induce RNAi in mammalian cells.
  • dsRNA molecules of 21 nucleotides in length are the most effective to induce RNAi.
  • dsRNA molecules effective to induce RNAi may be produced in vitro by solid phase synthesis.
  • dsRNA molecules may be produced in vitro by synthesis of sense and antisense strands from a DNA template using the T7 or a similar polymerase. Strands produced by this method can be annealed to form dsRNA molecules. dsRNA may also be produced in vivo by the production of short hairpin RNAs (shRNAs) from DNA vectors.
  • shRNAs short hairpin RNAs
  • short sequences of DNA complementary to the mRNA to be targeted are placed in an inverted repeat orientation under the control of a promoter.
  • the resulting transcript is self complementary and therefore folds back on itself to form a double-stranded hairpin-like structure.
  • the inverted repeat sequences may optionally be separated by a short heterologous DNA sequence. This sequence functions as a loop so that a hairpin-loop structure is formed by the transcript.
  • Rho GTPase expression can be determined at the level of mRNA or protein by any standard means well known in the art. Methods for determining mRNA levels include Northern blotting, RT-PCR, quantitative RT-PCR and RNAse protection assays. Methods for determining protein levels include Western blotting, immunohistochemistry and immunofluorescence.
  • Rho GTPases which are modulated by any of the above-mentioned means can include: GTP binding, GDP binding, GTPase activity, integrin binding, coupling or binding of Rho GTPases to receptor or effector-like molecules (such as integrins, growth factor receptors, tyrosine kinases, PI-3K, PIP-5K, etc.). Increasing, reducing, antagonizing, or promoting Rho GTPases can modulate the activity. Methods for determining the level of RJho GTPase activity in a sample are well known in the art. The level of activity of a Rho GTPase can be determined by measuring the amount of GTP binding.
  • Rho GTPase This is typically accomplished by labeling cells for a period of time with radioactively labeled inorganic phosphate, such as 32 P 5 which becomes incorporated into cellular GDP and GTP.
  • the Rho GTPase can then be immunoprecipitated using an appropriate antibody. Guanine nucleotides that are precipitated bound to the Rho GTPase can then be separated by thin layer chromatography and visualized by autoradiography. The ratio of GTP to GDP is indicative of the amount of Rho GTPase activity in the sample.
  • Rho GTPase activity in a sample is to use an affinity precipitation approach wherein the sample is incubated with an immobilized polypeptide that preferentially binds to the GTP-bound, versus GDP-bound, Rho GTPase of interest.
  • the amount of Rho GTPase that is precipitated by the immobilized polypeptide in a given sample can then be determined by standard means such as Western blotting.
  • the activity of Rac is frequently determined by incubating fresh cell lysate with the Rac-binding domain of the protein kinase PAK bound to agarose beads.
  • the agarose beads are then precipitated by centrifugation and the amount of bound Rac protein, which is indicative of the amount of GTP-bound Rac in the original sample, is determined by Western blotting.
  • compositions may be administered separate from an implanted material, either systemically or locally at the site of implantation, or as a component of the surface of an implanted material.
  • the compositions and functional derivatives can be formulated using standard techniques for enteral, parenteral, or topical administration. Effective dosages can be determined based on the in vitro assays known to those skilled in the art. In the case of proteins, one can also administer the compound by administering a nucleotide sequence encoding the compound, in a form that is expressed at a site in the patient.
  • Rho GTPase modulating agents may be further combined with other active agents.
  • Other useful agents include antiinflammatory agents.
  • Rho GTPase inhibitors may also be provided as a component of the surface layer of the material to be implanted. Rho GTPase inhibitors may be directly deposited onto a modified surface of the implanted material or may be included as a component of an additional outer surface layer of the implanted material.
  • the surface layer may be composed of any suitable material such as a porous matrix or a hydrogel.
  • the compositions described herein can be formulated for oral, intravenous or ophthalmic administration. Ophthalmic formulations can be applied topically to the eye, intraoccularly, or via the blood supply going to the eye.
  • Formulations are prepared using a pharmaceutically acceptable "carrier” composed of materials that are considered safe and effective and may be administered to an individual without causing undesirable biological side effects or unwanted interactions.
  • the “carrier” is all components present in the pharmaceutical formulation other than the active ingredient or ingredients.
  • carrier includes , but is not limited, to diluents, binders, lubricants, disintegrators, fillers, preservatives, buffering agents, surfactants, thickening agents, and coating compositions.
  • the formulation is preferably administered directly to the tissue or systemically via oral administration, injection (intravenous, intramuscular, or subcutaneous, pulmonary, nasal or transdermal methods.
  • the formulation may be administered as a solution, suspension, ointment, or other formulation for direct administration.
  • Injectable formulations are typically administered as solutions, suspension, or emulsions.
  • the formulations can be injected directly into the eye or into another part of the body.
  • the formulations can contain one or more pharmaceutically acceptable excipients such as surfactants, salts, buffers, pH modifying agents, emulsifiers, preservatives, anti-oxidants, osmolality/tonicity modifying agents, and water-soluble polymers.
  • suitable surfactants include, but are not limited, to those listed above.
  • Injectable formulations are typically buffered to a pH of 3-8 in order to avoid irritation at the site of injection.
  • Suitable buffers include, but are not limited to, phosphate buffers, acetate buffers, and citrate buffers.
  • Water soluble polymers are often used in formulations for parenteral administration. Suitable water-soluble polymers include, but are not limited to, polyvinylpyrrolidone, dextran, carboxymethylcellulose, and polyethylene glycol.
  • Preservatives can be used to prevent the growth of fungi and microorganisms.
  • Suitable antifungal and antimicrobial agents include, but are not limited to, benzoic acid, butylparaben, ethyl paraben, methyl paraben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetypyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, and thimerosal.
  • the formulation may be an immediate release or controlled release formulation, as described below.
  • Oral formulations can be administered in the form a tablet, capsule, solution, suspension, emulsion, syrup, etc.
  • the formulation will include excipients.
  • Diluents are typically necessary to increase the bulk of a solid dosage form so that a practical size is provided for compression of tablets or formation of beads and granules.
  • Suitable diluents include, but are not limited to, dicalcium phosphate dihydrate, calcium sulfate, lactose, sucrose, mannitol, sorbitol, cellulose, microcrystalline cellulose, kaolin, sodium chloride, dry starch, hydrolyzed starches, pregeiatinized starch, silicone dioxide, titanium oxide, magnesium aluminum silicate and powder sugar.
  • Binders are used to impart cohesive qualities to a solid dosage formulation, and thus ensure that a tablet or bead or granule remains intact after the formation of the dosage forms.
  • Suitable binder materials include, but are not limited to, starch, pregelatinized starch, gelatin, sugars (including sucrose, glucose, dextrose, lactose and sorbitol), polyethylene glycol, waxes, natural and synthetic gums such as acacia, tragacanth, sodium alginate, cellulose, including hydorxypropylmethylceilulose, hydroxypropylcellulose, ethylcellulose, and veegum, and synthetic polymers such as acrylic acid and methacrylic acid copolymers, methyl methacrylate copolymers, aminoalkyl methacrylate copolymers, polyacrylic acid/polymethacrylic acid and polyvinylpyrrolidone.
  • Lubricants are used to facilitate tablet manufacture of solid dosage forms.
  • suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, glycerol behenate, polyethylene glycol, talc, and mineral oil.
  • Disintegrants are used to facilitate dosage form disintegration or "breakup" after administration, and generally include, but are not limited to, starch, sodium starch glycolate, sodium carboxymethyl starch, sodium carboxymethylcellulose, hydroxypropyl cellulose, pregelatinized starch, clays, cellulose, alginine, gums or cross linked polymers, such as cross- linked polyvinylpyrrolidone (PVP) (Polyplasdone XL from GAF Chemical Corp). Stabilizers are used to inhibit or retard drug decomposition reactions which include, by way of example, oxidative reactions.
  • PVP polyvinylpyrrolidone
  • Stabilizers are used to inhibit or retard drug decomposition reactions which include, by way of example, oxidative reactions.
  • Preservatives are used to prevent the growth of fungi and microorganisms.
  • Suitable antifungal and antimicrobial agents include, but are not limited to, benzoic acid, butylparaben, ethyl paraben, methyl paraben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetypyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, and thimerosal.
  • Preservatives can also include antioxidants such a BHA, BHT, vitamin E, and other pharmacologically acceptable antioxidants.
  • suitable preservatives and their maximum concentrations include (a) benzalkonium chloride, 0.013%; (b) benzethonium chloride, 0.01%; (c) chlorobutanol, 0.5%; (d) phenylmercuric acetate, 0.004%; (e) phenylmercuric nitrie, 0.004%; and (f) thimerosal, 0.01%.
  • Surfactants may be anionic, cationic, amphoteric or nonionic surface active agents.
  • Suitable anionic surfactants include, but are not limited to, those containing carboxylate, sulfonate and sulfate ions.
  • anionic surfactants include sodium, potassium, ammonium of long chain alkyl sulfonates and alkyl aryl sulfonates such as sodium dodecylbenzene sulfonate; dialkyl sodium sulfosucchiates, such as sodium dodecylbenzene sulfonate; dialkyl sodium sulfosuccinates, such as sodium Ms-(2- ethylthioxyl)-sulfosuccinate; and alkyl sulfates such as sodium lauryl sulfate.
  • Cationic surfactants include, but are not limited to, quaternary ammonium compounds such as benzalkonium chloride, benzethonium chloride, cetrimonium bromide, stearyl dimethylbenzyl ammonium chloride, polyoxyethylene and coconut amine.
  • nonionic surfactants include ethylene glycol monostearate, propylene glycol myristate, glyceryl monostearate, glyceryl stearate, polyglyceryl-4-oleate, sorbitan acylate, sucrose acylate, PEG- 150 laurate, PEG-400 monolaurate, polyoxyethylene monolaurate, polysorbates, polyoxyethylene octylphenylether, PEG-1000 cetyl ether, polyoxyethylene tridecyl ether, polypropylene glycol butyl ether, Poloxamer 401, stearoyl monoisopropanolamide, and polyoxyethylene hydrogenated tallow amide.
  • Extended release formulations are generally prepared as diffusion or osmotic systems, for example, as described in "Remington— The Science and Practice of Pharmacy” (20th ed., Lippincott Williams & Wilkins, Baltimore, Md., 2000).
  • a diffusion system typically consists of two types of devices, reservoir devices and matrix devices, both of which are well known and described in the art.
  • the matrix devices are generally prepared by compressing the drug with a slowly dissolving polymer carrier into a tablet form.
  • the three major types of materials used in the preparation of matrix devices are insoluble plastics, hydrophilic polymers, and fatty compounds.
  • Plastic matrices include, but are not limited to, methyl acrylate-co-methyl methacrylale, polyvinyl chloride, and polyethylene.
  • Hydrophilic polymers include, but are not limited to, methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and
  • Fatty compounds include, but are not limited to, various waxes such as carnauba wax and glyceryl tristearate.
  • extended release formulations can be prepared using osmotic systems or by applying a semi-permeable coating to the dosage form.
  • the desired drug release profile can be achieved by combining low permeable and high permeable coating materials in suitable proportion.
  • the devices with different drug release mechanisms described above can be combined in a final dosage form comprising single or multiple units.
  • Examples of multiple units include multilayer tablets and capsules containing tablets, beads, granules, etc.
  • An immediate release portion can be added to the extended release system by means of either applying an immediate release layer on top of the extended release core using coating or compression processes or in a multiple unit system, such as a capsule, containing extended and immediate release beads.
  • Extended release tablets containing hydrophilic polymers are prepared by techniques commonly known in the art such as direct compression, wet granulation, or dry granulation processes. Their formulations usually incorporate polymers, diluents, binders, and lubricants as well as the active pharmaceutical ingredient.
  • the usual diluents include inert powdered substances such as any of many different kinds of starch, powdered cellulose, especially crystalline and microcrystal ⁇ ne cellulose, sugars such as fructose, mannitol and sucrose, grain flours and similar edible powders.
  • Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride and powdered sugar. Powdered cellulose derivatives are also useful.
  • Typical tablet binders include substances such as starch, gelatin and sugars such as lactose, fructose, and glucose. Natural and synthetic gums, including acacia, alginates, methylcellulose, and polyvinylpyrrolidine can also be used. Polyethylene glycol, hydrophilic polymers, ethylcellulose and waxes can also serve as binders.
  • a lubricant is necessary in a tablet formulation to prevent the tablet and punches from sticking in the die.
  • the lubricant is chosen from such slippery solids as talc, magnesium and calcium stearate, stearic acid and hydrogenated vegetable oils.
  • Extended release tablets containing wax materials are generally prepared using methods known in the art such as a direct blend method, a congealing method, and an aqueous dispersion method. In a congealing method, the drug is mixed with a wax material and either spray-congealed or congealed, screened, and processed.
  • Delayed release formulations are created by coating a solid dosage form with a film of a polymer which is insoluble in the acid environment of the stomach, and soluble in the neutral environment of small intestines.
  • the delayed release dosage units can be prepared, for example, by coating a drug or a drug-containing composition with a selected coating material.
  • the drug- containing composition may be, e.g., a tablet for incorporation into a capsule, a tablet for use as an inner core in a "coated core” dosage form, or a plurality of drug-containing beads, particles or granules, for incorporation into either a tablet or capsule.
  • Preferred coating materials include bioerodible, gradually hydrolyzable, gradually water-soluble, and/or enzymatically degradable polymers, and may be conventional "enteric" polymers.
  • Enteric polymers as will be appreciated by those skilled in the art, become soluble in the higher pH environment of the lower gastrointestinal tract or slowly erode as the dosage form passes through the gastrointestinal tract, while enzymatically degradable polymers are degraded by bacterial enzymes present in the lower gastrointestinal tract, particularly in the colon.
  • Suitable coating materials for effecting delayed release include, but are not limited to, cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl methyl cellulose acetate succinate, hydroxypropylmethyl cellulose phthalate, methylcellulose, ethyl cellulose, cellulose acetate, cellulose acetate phthalate, cellulose acetate trimellitate and carboxymethylcellulose sodium; acrylic acid polymers and copolymers, preferably formed from acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylate and/or ethyl methacrylate, and other methacrylic resins that are commercially available under the tradename EudragitTM (Rohm Pharma; Westerstadt, Germany), including EudragitTM L30D-55 and L1OO-55 (soluble at pH 5.5 and above), EudragitTM L-IOO (soluble
  • the preferred coating weights for particular coating materials may be readily determined by those skilled in the art by evaluating individual release profiles for tablets, beads and granules prepared with different quantities of various coating materials. It is the combination of materials, method and form of application that produce the desired release characteristics, which one can determine only from the clinical studies.
  • the coating composition may include conventional additives, such as plasticizers, pigments, colorants, stabilizing agents, glidants, etc.
  • a plasticizer is normally present to reduce the fragility of the coating, and will generally represent about 10 wt. % to 50 wt. % relative to the dry weight of the polymer.
  • plasticizers examples include polyethylene glycol, propylene glycol, triacet ⁇ n, dimethyl phthalate, diethyl phthalate, dibutyl phthalate, dibutyl sebacate, triethyl citrate, tributyl citrate, triethyl acetyl citrate, castor oil and acetylated monoglycerides.
  • a stabilizing agent is preferably used to stabilize particles in the dispersion.
  • Typical stabilizing agents are nonionic emulsif ⁇ ers such as sorbitan esters, polysorbates and polyvinylpyrrolidone. Glidants are recommended to reduce sticking effects during film formation and drying, and will generally represent approximately 25 wt. % to 100 wt.
  • glidant is talc.
  • Other glidants such as magnesium stearate and glycerol raonostearates may also be used.
  • Pigments such as titanium dioxide may also be used.
  • an anti-foaming agent such as a silicone (e.g., simethicone)
  • compositions can be applied topically as a spray, lotion, cream, gel, powder or ointment, using standard pharmaceutical carriers.
  • the compositions can also be administered as implants or as transdermal patches.
  • Suitable formulations are well known.
  • Ophthalmic preparations for topical administration are described in detail in "Pharmaceutical Dosage Forms and Drug Delivery Systems", by Anselm H. C, Popovich, N. G., and Allen, L. V., 6 th Ed., Williams and Wilkins, (1995).
  • Ophthalmic preparations are typically applied directly to the eye for the localized effect of the drug on the surface of the eye or on its interior.
  • Ophthalmic preparations are generally in the form of solutions or suspensions, but can also be formulated as ointments.
  • ophthalmic inserts impregnated with the active agent, can be used to provide continuous release of the agent. These inserts are particularly useful for agents requiring frequent daytime and nighttime administration.
  • Ophthalmic preparations are generally administered in small volumes due to the eye's limited capacity to retain liquids.
  • Ophthalmic suspensions are used to a lesser extent than ophthalmic solutions.
  • Suspensions can be used to increase the corneal contact time of the drug and thus provide a more sustained action. Suspensions can also be used when the drug is insoluble in the desired vehicle or unstable in solution form. Suspensions must contain particles of such chemical characteristics and dimensions that they are non- irritating to the eyes. Suspensions can also include dispersing agents which prevent the agglomerization of the suspended particles. Suspensions are typically shaken immediately prior to use to uniformly distribute the drug particles throughout the vehicle.
  • Ophthalmic solutions/suspensions are typically administered using a dropper.
  • the patient uses an index finger to gently pull down the affected eye to form a pocket or cup. Without touching the dropper to the eye, the prescribed number of drops are placed into the formed pocket. Direct administration onto the cornea is not recommended as it may cause the corneal reflex to initiate a blink and thus lessen the amount of drug available for absorption into the eye.
  • the patient should press a finger against the inner corner of the eye for approximately one minute. This prevents the medication from entering the tear duct and lessens the amount of drug entering the nose, the gastrointestinal tract, and ultimately the systemic circulation.
  • the attending physician will prescribe for each patient the best number of drops to apply, the frequency of application, and the duration of treatment necessary.
  • Buffering agents may be used in ophthalmic solutions/suspensions to reduce discomfort to the patient, ensure drug stability, and to control the therapeutic activity of the drug. Although the patient's natural tears are normally sufficient to buffer the ophthalmic solution/suspension, buffering agents such as phosphates can also be used. Formulations typically are buffered to a pH of between about 6 and about 8. Buffer salts such as phosphates, borates, and citrates are chosen based on their compatibility with the drug or combination drug to be administered. Thickening agents can be used to increase the viscosity of ophthalmic solutions/suspensions, which can increase the retention time of the formulation, decrease the drainage rate, and increase ocular bioavailability.
  • Suitable thickening agents include, but are not limited to, methylcellulose, hydroxypropyl cellulose, and polyvinyl alcohol.
  • Osmolarity-adjusting agents can be used in order to improve patient comfort.
  • Suitable osmolarity-adjusting agents include, but are not limited to, sodium chloride and mannitol.
  • Antimicrobial preservatives can be used in ophthalmic solutions/suspensions. Suitable preservatives include, but are not limited to, benzalkonium chloride, thimerasol, chlorobutanol, parabens such as methyl and propyl paraben, and polyquat (polyquaternium-1). Antioxidants can be used in ophthalmic solutions/suspensions.
  • Suitable antioxidants include, but are not limited to, sodium metabisulfite or metabisulfite.
  • Ophthalmic ointments unlike dermatological ointments, must be sterile. Ophthalmic ointments are either prepared from sterile ingredient under aseptic conditions or are sterilized following manufacture.
  • the ointment base must be non-irritating to the eye and must permit diffusion of the drug throughout the secretions covering the eye.
  • bases typically have a softening point or melting point close to body temperature. Suitable bases include, but are not limited to, mixtures of petrolatum and liquid petrolatum and mixtures of mineral oil and white petrolatum. Water misc ⁇ ble agents such as lanolin may be added to the formulation.
  • the drug is added to the ointment base either as a solution or as a finely micronized powder. The drug is then mixed well with the base, usually by milling. The ointment is applied as a thin ribbon in the lower lid of the eye. The attending physician will advise the patient with respect to the frequency of application and the duration of treatment necessary.
  • Formulations allowing for prolonged residence times on the surface of the eye, thus increasing bioavailability of the drug, use a base composed of C ARBOMER®, which is a high molecular weight, cross-linked acrylic acid copolymer having a high viscosity.
  • the formulation typically contains a relatively low concentration of the polymer (about 5%, with the remainder of the gel base being water).
  • An example of such a formulation is Pilopine HS gel 24 which contains policarpine HCl for the treatment of glaucoma.
  • Intraocular formulations are used to administer drugs within the eye.
  • examples of such formulations include irrigation solutions used during eye surgery and viscoelastic agents, which are used during surgery to protect the cornea endothelium (e.g. Healon-sodium hyaluronate from Pharmacia).
  • Drugs can also be administered using ocular inserts or implants.
  • the inserts or implants can be erodible or non-erodible.
  • Alza developed a non- erodible controlled release topical dosage form in 1973 known as the Ocusert system.
  • the system was a soft, flexible elliptical membrane designed to be placed in the cul-de-dac of the eye.
  • the dosage form contained a drug containing core sandwiched between two copolymer membranes.
  • the system was used for the administration of pilocarpine over seven days for the treatment of glaucoma (Ocusert PiIo)
  • Lacrisert is a sterile, translucent, rod-shaped, water soluble ophthalmic insert made of hydroxypropyl cellulose ("HPC") for administration into the inferior cul-de-dac of the eye developed by Alza Corp.
  • HPC hydroxypropyl cellulose
  • the insert is designed to release the drug via controlled erosion of the HPC matrix (i.e. degradation), not diffusion.
  • the insert can be used to deliver poorly soluble, high molecular weight compounds.
  • the insert is composed of a drug reservoir and an annular ring sandwiched between two rate controlling membranes. The rate controlling membranes absorb water to form a gel-like mass which erodes over time and releases the active agent.
  • the materials can be applied to the surface of, incorporated into, or administered with, implants, electrodes, sensors, stents, catheters, tissue engineering devices, pumps including intrathecal pumps, prosthetics or parts of prosthetics (screws, rivets, etc.), periodontal implants and other devices.
  • the materials may be metal, ceramic, polymeric, bone, hydroxyapatite, or other materials commonly used in such devices.
  • IL Methods to inhibit foreign body giant cell formation include contacting cells with one or more inhibitors of Rho family GTPases.
  • the methods include contacting an implanted material or the adjacent tissue or bodily fluids with an effective amount of a Rho GTPase antagonist to prevent formation of multinucleated foreign body giant cells at the implantation site.
  • Inhibitors of Rho GTPase activity may be provided separately from the implanted material, or alternatively, may be provided as a component of the external surface of the implanted material.
  • Methods for inhibiting foreign body giant cell formation may be used, for example, to reduce surface damage to implanted materials and enhance the biocompatibility of an implanted material.
  • the methods may be used in any situation in which it is desirable to reduce macrophage fusion and/or encapsulation at the site of the implant, thereby prolonging the working lifetime and efficiency of the implanted device.
  • the described methods can be used to enhance biocompatibility of devices that are completely implanted into a living body (i.e., the entire device is implanted within a living body).
  • cardiovascular devices such as vascular grafts and stents
  • artificial blood vessels such as vascular grafts and stents
  • prosthetic devices such as artificial hip joints and artificial knee joints
  • scaffolds that support tissue growth in such anatomical structures as bone, tooth, nerves, pancreas, eye and muscle.
  • the methods are also useful for enhancing biocompatibility of devices that are partially implanted within a living body (i.e., only part of the device is implanted within a living body, the remainder of the device being located outside of the body).
  • partially implantable medical devices include, but are not limited to, biosensors (such as those used to monitor the level of drugs within a living body, or the level of blood glucose in a diabetic patient), percutaneous devices (such as catheters) that penetrate the skin and link a living body to a medical device such as a kidney dialysis machine, and skin substitutes (such as dermal and epidermal scaffolds).
  • biosensors such as those used to monitor the level of drugs within a living body, or the level of blood glucose in a diabetic patient
  • percutaneous devices such as catheters
  • a medical device such as a kidney dialysis machine
  • skin substitutes such as dermal and epidermal scaffolds
  • An inhibition of the FBR at the site of implantation can be identified, for example, by at least one of the following changes: a decrease in persistence of inflammatory cells (such as foreign body giant cells and/or activated macrophages) present at the site of implantation; a decrease in the level of cytokines, such as interleukin and monocyte chemoattractant protein in extracts of tissue taken from the site of implantation by ELISA; a decrease in the growth factors secreted by cells, such as TGF-beta; or a decrease in the amounts of proteolytic enzymes such as matrix metalloproteinases, collagenases, elastases and acid hydrolases (measured, for example, by analyzing tissue extracts by zyrnography, Western blot, ELISA, or immunohistochemical staining of tissue sections). Tissue extracts can also be analyzed by the PAI-I lucifierase assay using mink lung epithelial cells (Kyriakides, et al, Am, J.
  • An inhibition of encapsulation at the site of implantation can be characterized, for example, by at least one of the following: a decrease in the amount of Fibrosis (measured, for example, by determining the level of hydroxyproline content which indicates the level of collagen in the foreign body capsule); a decrease in capsule thickness (measured, for example, by examination of histological sections with the aid of an ocular micrometer); a decrease in the amount of contraction of collagen fibers within the capsule (measured, for example, as tensile strength of the capsule or induced shape change on malleable implants); or a decrease in the diffusion rates of small molecules through the capsule (measured, for example, as described by Sharkawy, et al.J. Biomed Mater. Res. t 37:401-412 (1997)).
  • Example 1 IL-4 and GM-CSF Induce Fusion of Marrow-Derived Murine Monocytes.
  • Mouse monocytes were obtained from the bone marrow of C57BL6 mice (Jackson Laboratories). Monocytes derived from mice that lack MCP-I (93% C57BL6, 7% SJL) were used as a negative control for fusion. Marrow was flushed from the femurs of mice and collected in Iscove's Modified Dulbecco's Medium (IMDM) (Gibco) supplemented with 10% FBS and penicillin/streptomycin/fungizone (PSF). Cells were layered over Lympholyte-M (Accurate Chemical) and centrifuged.
  • IMDM Iscove's Modified Dulbecco's Medium
  • PSF penicillin/streptomycin/fungizone
  • the traction containing mononuclear cells was collected and plated (2.5 X lO 6 cells/ml) in expansion medium (IMDM supplemented with 20% FBS, 1-glutamine, PSF, and 1.5 ng/ml huM-CSF (R&D Systems), and 100 ng/ml flt-3 ligand (R&D Systems). Cultures were fed on day 5 and cells were collected by scraping on day 10 and used for analysis.
  • IMDM IMDM supplemented with 20% FBS, 1-glutamine, PSF, and 1.5 ng/ml huM-CSF (R&D Systems), and 100 ng/ml flt-3 ligand (R&D Systems).
  • Expanded monocytes were plated at 1 X 10 6 cells per well in non- tissue culture treated polystyrene 24 well plates in expansion media lacking M-CSF and flt-3 ligand and supplemented with 10 ng/ml recombinant mouse GM-CSF and 10 ng/ml recombinant mouse IL-4. Media was changed every 3 days for the duration of all experiments involving cell lines and at days 3 and 5 for all experiments involving mouse monocytes.
  • monocytes from mice that lack MCP-I were prepared and examined for cell fusion. Previous results have shown that FBGC formation in vivo was compromised in these mice and that inhibition of MCP-I activity limited the fusion of human monocytes (Kyriakides, etal., Am. J.
  • Monocytes were obtained from the bone marrow of C57BL6 mice, cultured, and induced to fuse by the concurrent addition of GM-CSF and IL- 4 as in Example 1.
  • GM-CSF GM-CSF
  • IL- 4 IL- 4
  • cells were fixed in 4% paraformaldehyde (JT Baker) for 20 minutes at RT and stained with rhodamine-Phalloidin and DAPI according to standard protocols. All wells were mounted with Vectashield (Vector Labs) and examined with the aid of an Axiovert 200M Zeiss microscope equipped with fluorescent optics.
  • Cells were induced to fuse by the concurrent addition of GM-CSF and ⁇ L-4 in the presence of the small molecular weight inhibitor of Racl activation, NSC23766, to test for the involvement of this GTPase in the cell fusion process.
  • NSC23766 small molecular weight inhibitor of Racl activation
  • cells were double-stained with May-Grunwald stain (Sigma) and Wright- Giemsa stain (Sigma) according to standard protocols.
  • For visualization of the actin cytoskeleton and nuclei cells were fixed in 4% paraformaldehyde (JT Baker) for 20 minutes at RT and stained with rhodamine-Phalloidin and DAPI according to standard protocols. All experiments were done in triplicate and a total of fifteen high power fields were analyzed for each group.
  • FBGC Foreign body giant cell
  • Monocytes were obtained from the bone marrow of C57BL6 mice, and cultured as in Example 1. Cells were induced to fuse by the concurrent addition of GM-CSF and IL-4 in the presence of the small molecular weight inhibitor of Racl activation, NSC23766, and in the presence of polystyrene Fluoresbrite YG microspheres with a 3 ⁇ M diameter (Polysciences). To visualize microspheres, the actin cytoskeleton and nuclei, cells were fixed in 4% paraformaldehyde (JT Baker) for 20 minutes at RT and stained with rhodamine-Phalloidin and DAPI according to standard protocols. All wells were mounted with Vectashield (Vector Labs) and examined with the aid of an Axiovert 200M Zeiss microscope equipped with fluorescent optics. Results:
  • NSC23766 to monocyte cultures that were induced to fuse in the presence of polystyrene microspheres for 7 days indicated that even though fusion was limited, the uptake of spheres was not compromised. Phagocytosis was not inhibited, as few non-engulfed microspheres were observed. In the absence of the Racl inhibitor, giant cell formation and uptake of beads were evident. However, addition of NSC23766 limited fusion without limiting the ability of monocytes to uptake spheres. As expected, limited lamellipodia formation was observed in NSC23766 treated cells, indicating a potential role for lamellipodia formation in fusion.
  • the dried EVAc was then cut into small disks ( ⁇ 3 mm diameter, ⁇ 2 mm thickness, 11.19 +/- 1.07 mg) with a cork borer and sterilized under UV light for 20 minutes. Control disks were made with 100 mg Ficoll. Implants were excised en bloc, at 2 weeks following implantation, as described previously and processed for histological and immunohistochemical analysis (Kyriakides, et ah, 2004). Briefly, explanted biomaterials were fixed in 1% zinc-buffered formalin for 24 hours. Following processing and embedding, 6 ⁇ M sections were stained with hematoxylin and eosin (H&E) or were processed for immunohistochemistry with Mac3 antibody (1 :500 dilution). Mac3 localization was determined using a secondary peroxidase-conjugated antibody and visualization of peroxidase activity using hydrogen peroxide and DAB 3,3'- diaminobenzidine which forms an insoluble brown-colored polymer when oxidized.
  • NSC23766 ⁇ eluting EVAc scaffolds were prepared and the release of the drug was evaluated in vitro and determined to be in the range of 5 ⁇ g/mg of scaffold and was sustained for at least 18 days.
  • EVAc scaffolds with or without NSC23766 were implanted SC in wild type mice for 2 weeks and the formation of FBGC was evaluated. A significant reduction in the number of FBGC surrounding NSC23766-elutmg implants in comparison to control implants was observed (Figure 4). Macrophage recruitment, assessed by immunohistochemistry with Mac3 antibody, and the overall foreign body reaction, assessed by H&E staining, was similar between the two groups.

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Abstract

L'invention porte sur des procédés et des compositions qui permettent d'inhiber la réponse à corps étranger face à des matériaux implantés. Les procédés consistent à inhiber la formation de cellules géantes à corps étranger en administrant une quantité efficace d'un antagoniste de l'activité GTPasique de la Rho. Dans un mode de réalisation préféré, les procédés consistent à mettre en contact un matériau implanté ou le tissu ou les liquides corporels adjacents avec une quantité efficace d'un antagoniste de l'activité GTPasique de la Rho afin d'empêcher la formation de cellules géantes multinucléées à corps étranger sur le site d'implantation. Les inhibiteurs de l'activité GTPasique de la Rho peuvent être fournis séparément du matériau implanté ou, dans un autre mode de réalisation, peuvent être fournis sous la forme d'un composant de la surface externe du matériau implanté. Les procédés de l'invention consistent à mettre en contact des monocytes ou des macrophages avec des inhibiteurs de l'activité GTPasique de la famille des Rho dans des quantités efficaces pour inhiber la fusion desdites cellules, fusion qui débouche sur la création de cellules géantes multinucléées à corps étranger. De préférence, les inhibiteurs de l'activité GTPasique de la Rho sont des inhibiteurs de l'activité GTPasique de la famille des Rac. Dans un mode de réalisation préféré, on utilise un inhibiteur Rac à petites molécules NSC23766.
PCT/US2007/085598 2006-11-27 2007-11-27 Procédés permettant d'inhiber la réaction à corps étranger face à des matériaux implantés WO2008067288A2 (fr)

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WO2012069193A1 (fr) * 2010-11-26 2012-05-31 Reza Ghotbi Implant pour le traitement ou la prévention d'un anévrisme
WO2022076700A1 (fr) * 2020-10-07 2022-04-14 The Board Of Trustees Of The Leland Stanford Junior University Procédés et compositions de traitement de complications associées à un implant
WO2024049713A1 (fr) * 2022-08-30 2024-03-07 Lintec Of America, Inc. Support de régénération nerveuse améliorée pour une repousse accélérée
WO2024121426A1 (fr) 2022-12-09 2024-06-13 Cemm - Forschungszentrum Für Molekulare Medizin Gmbh Procédé d'optimisation de lymphocytes t pour une immunothérapie

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WO2003105923A2 (fr) * 2002-06-14 2003-12-24 The Trustees Of Columbia University In The City Ofnew York Utilisation de y-27632 comme agent destine a prevenir la restenose apres angioplastie d'arteres coronaires/implantation d'endoprothese

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012069193A1 (fr) * 2010-11-26 2012-05-31 Reza Ghotbi Implant pour le traitement ou la prévention d'un anévrisme
CN103298500A (zh) * 2010-11-26 2013-09-11 雷扎·古特比 用于治疗或预防血管瘤的植入物
WO2022076700A1 (fr) * 2020-10-07 2022-04-14 The Board Of Trustees Of The Leland Stanford Junior University Procédés et compositions de traitement de complications associées à un implant
WO2024049713A1 (fr) * 2022-08-30 2024-03-07 Lintec Of America, Inc. Support de régénération nerveuse améliorée pour une repousse accélérée
WO2024121426A1 (fr) 2022-12-09 2024-06-13 Cemm - Forschungszentrum Für Molekulare Medizin Gmbh Procédé d'optimisation de lymphocytes t pour une immunothérapie

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