WO1996035418A1 - Procede d'inhibition de la liaison de selectines aux sialyl-lewis - Google Patents

Procede d'inhibition de la liaison de selectines aux sialyl-lewis Download PDF

Info

Publication number
WO1996035418A1
WO1996035418A1 PCT/US1996/006453 US9606453W WO9635418A1 WO 1996035418 A1 WO1996035418 A1 WO 1996035418A1 US 9606453 W US9606453 W US 9606453W WO 9635418 A1 WO9635418 A1 WO 9635418A1
Authority
WO
WIPO (PCT)
Prior art keywords
selectin
cells
lewis
binding
sialyl
Prior art date
Application number
PCT/US1996/006453
Other languages
English (en)
Inventor
Timothy P. Kogan
Ian L. Scott
Karin M. Keller
Brian Dupre
Robert J. Bjercke
Robert V. Market
Sidney J. Sherwood
Edward T. Yeh
Ronald G. Tilton
Phi Nga Thi Kint
Huong M. Bui
Pamela J. Beck
Original Assignee
Texas Biotechnology Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Texas Biotechnology Corporation filed Critical Texas Biotechnology Corporation
Priority to AU57322/96A priority Critical patent/AU5732296A/en
Publication of WO1996035418A1 publication Critical patent/WO1996035418A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/194—Carboxylic acids, e.g. valproic acid having two or more carboxyl groups, e.g. succinic, maleic or phthalic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]

Definitions

  • This invention relates to methods of
  • E-selectin which has also been called ELAM- 1 for endothelial leukocyte adhesion molecule-1 and
  • LECAM-2 for lectin cell adhesion molecule is a
  • endothelial cells the cells that line the interior wall of capillaries.
  • E-selectin recognizes and binds to the carbohydrate sialyl-Lewis x (sLe x ), which is present on the surface of certain white blood cells.
  • E-selectin helps white blood cells recognize and adhere to the capillary wall in areas where the tissue
  • E-selectin is actually one of three selectins now
  • P-selectin is expressed on activated or inflamed
  • sialyl-Lewis x sialyl-Lewis a (sLe a ), Lewis x and Lewis a are shown below.
  • white blood cells also called leukocytes
  • a tissue has been invaded by a microorganism or has been damaged
  • white blood cells also called leukocytes
  • P- and E-selectin have been shown to bind to specific types of white blood cells and enable these cells to recognize the affected sites and bind to the capillary wall so that these white blood cells may diffuse into the affected tissue.
  • L-selectin is present on the surface of most leukocytes and has been shown to recognize and bind to sLe x -related epitopes present on the surface of activated endothelial cells and high endothelial venules. L-selectin has also been shown to bind to specific types of endothelial cells and to enable leukocytes to be localized at these sites so that they may then migrate from the blood vessel into the
  • E-selectin and P-selectin recognize sLe x and related oligosaccharides presented as
  • glycoproteins or glycolipids on the surface each of these types of cells including neutrophils, monocytes, a specific subset of T lymphocytes, eosinophils and basophils.
  • neutrophils include monocytes, a specific subset of T lymphocytes, eosinophils and basophils.
  • Neutrophils are a subclass of granulocytes that phagocytose and destroy small organisms,
  • Monocytes after leaving the bloodstream through the wall of a capillary, mature into macrophages that phagocytose and digest invading microorganisms, foreign bodies and senescent cells. Lymphocytes produce antibodies and kill infected cells. Eosinophils and basophils secrete mediators of various inflammatory reactions.
  • Monocytes and neutrophils are able to recognize the site where a tissue has been damaged by binding to E-selectin or P-selectin, which is produced on the surface of the endothelial cells lining
  • E-selectin and P-selectin is increased when the tissue adjacent a capillary is affected.
  • P-selectin is present constitutively in storage granules from which it can be rapidly mobilized to the cell surface after the endothelium has been activated.
  • E- selectin requires de novo RNA and protein synthesis, and peak expression does not occur until about 4-6 hours after activation, and declines to basal levels after about 24-48 hours.
  • Leukocytes are able to migrate into affected areas because sLe x or sLe a moieties present on the surface of the leukocytes bind to E-selectin, P-selectin and/or L-selectin present on the surface of endothelial cells. This binding slows the flow of white blood cells through the bloodstream, since it mediates the rolling of leukocytes along the activated endothelium prior to integrin mediated attachment and migration, and helps to localize white blood cells in areas of injury or infection.
  • sLe a is found on various cancer cells, including lung and colon cancer cells. It has been suggested that cell adhesion involving sLe a may be involved in the metastasis of certain cancers and that inhibitors of sLe a binding may be useful in the treatment of some forms of cancer.
  • This invention provides a method of inhibiting the binding of E-selectin, P-selectin and/or L-selectin to sialyl-Lewis x , sialyl-Lewis a ' Lewis x and/or Lewis a presented on a cell surface comprising the step of administering to a patient in need of such treatment a pharmaceutically effective amount of at least one compound having the formula
  • R 1 is an alkyl group, an alkoxy group or a halogen atom
  • R 2 is a halogen atom or a hydrogen atom
  • R 3 is an alkyl group or a hydrogen atom
  • the compounds useful in the method of the present invention may be used for treating diseases such as septic shock, reperfusion injury that occurs following heart attacks, strokes and organ transplants, traumatic shock, multi-organ failure, autoimmune diseases, asthma, inflammatory bowel disease, Crohn's disease, and ARDS, in a method which comprises administering to a patient suffering from any of the diseases a
  • Fig. 1 is a graph that measures bronchial responsiveness by comparing the specific lung
  • Fig. 2 is a graph that measures bronchial responsiveness by determining the cummulative carbochol concentration in breath units for compound 2 of the present invention.
  • R 1 is an alkyl group, an alkoxy group or a halogen atom
  • R 2 is a halogen atom or a hydrogen atom
  • R 3 is an alkyl group or a hydrogen atom
  • R 1 is as defined above
  • AA is an ⁇ -amino acid
  • R 1 and R 2 are as defined above.
  • R 1 is an alkyl group and R 2 is a hydrogen atom
  • R 1 is an alkoxy group and R 2 is a hydrogen atom or a halogen atom
  • R 1 is a halogen atom and R 2 is a hydrogen atom.
  • the compounds of the present invention can be synthesized using methods well known to those skilled in the art. See, for example, Collect. Czech. Chem. Comm., 51(11), pp. 2617-25, 1986 (English) or U.S. Patent Nos. 3,904,682 and 4,009,197.
  • alkyl shall mean a monovalent straight chain or branched chain group of 1 to 12 carbon atoms including, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl and the like.
  • lower alkyl shall mean any alkyl group having from one to six carbon atoms.
  • halogen shall mean any atom selected from the group consisting of chlorine, fluorine, bromine, and iodine.
  • alkoxy shall mean an alkyl group attached to a molecule through an oxygen atom including, but not limited to, methoxy, ethoxy, isopropoxy,
  • salts refers to those carboxylate salts, amino acid addition salts, esters, amides and prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of patients without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
  • salts refers to the relatively non-toxic, inorganic and organic acid addition salts of the compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds or by separately reacting the purified compound in its free base form with a suitable organic or
  • Representative salts include cations based on the alkali and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium and the like, as well as nontoxic ammonium, quaternary ammonium and amine cations including, but not limited to, ammonium,
  • esters of the compounds of this invention include C 1 to C 6 alkyl esters wherein the alkyl group is a straight or branched chain. Acceptable esters also include C 5 to C 7 cycloalkyl esters as well arylalkyl esters such as, but not limited to benzyl. C 1 to C 4 a4kyl esters are preferred. Esters of the compounds of the present invention may be prepared according to
  • Examples of pharmaceutically acceptable, nontoxic amides of compounds of this invention include amides derived from ammonia, primary C 1 to C 6 alkyl amines and secondary C 1 to C 6 dialkyl amines wherein the alkyl groups are straight or branched chain. In the case of secondary amines the amine may also be in the from of a 5 or 6 membered heterocycle containing one nitrogen atom. Amides derived from ammonia, C 1 to C 3 alkyl primary amides and C 1 to C 2 dialkyl secondary amides are
  • Amides of the compounds of the invention may be prepared according to conventional methods.
  • prodrug refers to compounds that are rapidly transformed in vivo to yield to the parent compound of the above formula, for example by hydrolysis in blood.
  • a thorough discussion is provided in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems", Vol 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference.
  • compositions that include one or more of the compounds having the above structures formulated together with one or more nontoxic, physiologically acceptable carriers, adjuvants or vehicles, which are collectively referred to herein as carriers, for
  • parenteral injection for oral administration in solid or liquid form, for rectal or topical administration and the like.
  • compositions can be administered to humans and animals either orally, rectally, parentally
  • compositions suitable for parenteral injection may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions.
  • suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol,
  • polyol (propylene glycol, polyethylene glycol, glycerol and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate.
  • vegetable oils such as olive oil
  • injectable organic esters such as ethyl oleate.
  • Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants.
  • compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.
  • the compounds can be incorporated into slow or timed release or targeted delivery systems such as polymer matrices, liposomes, and microspheres. They may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating
  • sterilizing agents in the form of sterile water, or some other sterile injectable medium immediately before use.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.
  • the active compound or a pro- drug ester is admixed with at least one inert customary excipient (or carrier) such as sodium citrate or
  • dicalcium phosphate or (a) fillers or extenders, as for example, starches, lactose, sucrose, glucose, mannitol and silicic acid, (b) binders, as for example,
  • the dosage forms may also comprise buffering agents.
  • humectants as for example, glycerol
  • diintegrating agents as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and sodium carbonate
  • e solution retarders, as for example, paraffin
  • absorption accelerators as for example, quaternary ammonium compounds
  • wetting agents as for example, cetyl alcohol and glycerol monostearate
  • adsorbents as for example, kaolin and bentonite
  • lubricants as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate or mixtures thereof.
  • the dosage forms may also comprise buffering agents.
  • compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with
  • coatings and shells such as enteric coatings and others well known in the art. They may contain opacifying agents, and can also be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner.
  • embedding compositions examples include polymeric substances and waxes.
  • the active compounds can also be in
  • microencapsulated form if appropriate, with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art such as water or other solvents, solubilizing agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl
  • oils in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil and sesame seed oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan or mixtures of these substances, and the like.
  • compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.
  • Suspensions in addition to the active compounds, may contain suspending agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth or mixtures of these substances and the like.
  • suspending agents as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth or mixtures of these substances and the like.
  • compositions for rectal administrations are preferably suppositories, which can be prepared by mixing the compounds of the present invention with suitable nonirritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ordinary temperatures but liquid at body temperature and therefore melt in the rectal or vaginal cavity and release the active component.
  • suitable nonirritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ordinary temperatures but liquid at body temperature and therefore melt in the rectal or vaginal cavity and release the active component.
  • Dosage forms for topical administration of a compound of this invention include ointments, powders, sprays and inhalants.
  • the active component is admixed under sterile conditions with a physiologically acceptable carrier and any needed preservatives, buffers or propellants as may be required.
  • a physiologically acceptable carrier and any needed preservatives, buffers or propellants as may be required.
  • Ophthalmic formulations, eye ointments, powders and solutions are also contemplated as being within the scope of this invention.
  • liposomes are generally derived from phospholipds or other lipid substances. Liposomes are formed by mono or multilamellar hydrated liquid crystals that are dispersed in an aqueous medium. Any nontoxic, physiologically acceptable and metabolizable lipid capable of forming liposomes can be used.
  • the present compositions in liposome form can contain, in addition to the selectin binding inhibitors of the present invention, stabilizers, preservatives, excipients and the like.
  • the preferred lipids are the phospholipids and the
  • phosphatidyl cholines both natural and synthetic.
  • Methods to form liposomes are well known in the art.
  • compositions of the present invention may be varied so as to obtain an amount of active ingredient that is effective to obtain the desired therapeutic response for a particular composition and method of administration.
  • the selected dosage levels therefore, depends on the desired therapeutic effect, on the route of
  • the total daily dosage of the compounds of this invention administered to a host in single or divided doses may be in the range of from about 5 mg to about 250 mg per kilogram of body weight.
  • Dosage unit compositions may contain such submultiples thereof as may be used to make up the daily dosage. It will be understood, however, that the specific dose level for any particular patient, whether human or other animal, will depend upon a variety of factors including the body weight, general health, sex, diet, time and route of administration, rates of absorption and excretion, combination with other drugs and the severity of the particular disease being treated.
  • the compounds useful in the methods of the present invention may be used to treat a variety of diseases relating to cell-cell recognition and adhesion.
  • the compounds of the present invention may be administered to a patient to treat septic shock, and reperfusion tissue injury that occurs following heat attacks, strokes and organ transplants, traumatic shock, multi-organ failure, autoimmune
  • an effective amount of the compounds useful in the present invention is administered either alone or as part of a pharmaceutically active composition to a patient in need of such treatment. It is also recognized that a combination of the compounds may be administered to a patient in need of such administration.
  • the compounds of the present invention may also be administered to treat other diseases that are associated with cell-cell
  • any disease that is related to this interaction may potentially be treated by the inhibition of this binding interaction.
  • the E-selectin binding assays involved assessing the ability of HL60 cells that express sialyl-Lewis x and Lewis x to bind cells expressing E-selectin (cell-cell assay) or to bind to purified E-selectin recombinant protein (cell-protein assay). Both P-selectin and L- selectin binding was assessed using a cell-protein assay. A similar binding assay utilizing purified glycolipids and purified L-selectin recombinant protein (glycolipid- protein) was used to assess L-selectin binding.
  • a human E-selectin cDNA was amplified by the polymerase chain reaction (PCR) from cDNA purchased from R&D Systems (Minneapolis, MN) .
  • PCR polymerase chain reaction
  • the E-selectin cDNA was ligated into a mammalian expression vector and
  • ECV umbilical vein endothelial cell-derived cell line
  • E-selectin expressing ECV cells and nontransfected ECV cells were plated in 24 well cluster dishes at a density of 10 5 cells/well 24-48hr prior to assay. The dishes were washed with binding buffer (RPMI, 1% BSA) immediately prior to assay. HL60 cells were
  • Non idet 40 (NP40) detergent containing 1% Non idet 40 (NP40) detergent
  • mice were PCR amplified cDNA generated from RNA extracted from the hybridoma cell line 402C10. All fusion cassettes were expressed from baculovirus vectors using the BakPAK method and SF21 cells purchased from Clonetech.
  • Recombinant fusion proteins were purified from baculovirus infected culture supernatants by
  • beads incubated with mock culture supernatants did not bind HL60 cells that express sialyl- Lewis x served as a negative control.
  • Beads incubated from E-, L- or P-selectin culture supernatants did bind HL60 cells.
  • HL60 cells (10 7 cells) were fluorescently labeled with calcein AMC-3099 (Molecular Probes) in RPMI 1640 with 10% fetal calf serum (FCS).
  • FCS fetal calf serum
  • Cytofluor 2350 fluorimeter Dose response curves and the concentration of compound at which 50% of the cell binding was inhibited (IC50) values were determined.
  • mice were PCR cloned from total RNA extracted from human placenta.
  • the mouse IgG cDNA was cloned from PCR amplified cDNA generated from RNA
  • Recombinant fusion proteins were purified from baculovirus infected culture supernatants by
  • beads incubated with mock culture supernatants did not bind glycolipids purified from HL60 cells that express sialyl-Lewis x and served as a negative control.
  • Beads incubated from E-, L- or P- selectin culture supernatants did bind glycolipids extracted from HL60 cells.
  • Glycolipids extracted from HL60 cells were coated onto 96 well flexible assay plates. Plates were
  • BSA bovine serum albumin
  • the ELISA absorbance 415 readings were curve fit to serially diluted mock and E-selectin IgG coated bead standards, both of which had essentially identical amounts of goat IgG. Dose response curves and the concentration of compound at which 50% of the cell binding was inhibited (IC50) values were determined.
  • PBS Dulbecco's phosphate buffered saline
  • Control animals receive 1ml of PBS alone.
  • Each dose of 0.1 ml of 50mg/ml solutions administered in PBS at neutral pH of the compound of interest is injected subcutaneously at: -15 minutes, time 0, +15 minutes, +45 minutes, and +75 minutes.
  • Mice are sacrificed at 150 or 180 minutes and a 10ml peritoneal lavage composed of PBS, EDTA, BSA, and gelatin is performed.
  • the total number of cells in the exudate is determined using a hemocytometer and the number of polymorphonuclear leukocytes (PMN's) is determined using cytospin preparations treated with
  • Lipopolysaccharide 50 ⁇ l, (1 ⁇ g/ ⁇ l) was injected into the hind footpad of 200 gram Lewis rats. Control rats received an equal volume of 0.9% saline. Compound 2, 0.5ml, (150mM concentration) was injected
  • Aqueous fluid was collected from both eyes of rats using a 100 ⁇ l heparinized capillary tube 24 hours after the initial injection of lipopolysaccharide. Aliquots of the aqueous fluid were subsequently spread on a
  • a Balloon catheter was advanced through one nostril into the lower esophagus.
  • the animals were intubated with a cuffed endotracheal tube through the other nostril with flexible fiber optic bronchoscope as a guide.
  • Pleural pressure was estimated with the esophageal balloon catheter (filled with 1 ml of air), which was positioned 5-10 cm from the gastroesophageal junction. In this position the end expiratory pleural pressure ranged between -2 and -5 cm H 2 O. Once the balloon was placed, it was secured so that it remained in the same position for the duration of the experiment. Lateral pressure in the trachea was measured with a sidehole catheter (inner diameter, 2.5 mm) advanced through and positioned distal to the tip of the endotracheal tube. The tracheal and pleural pressure catheters were
  • transpulmonary pressure which was defined as the difference between tracheal and pleural pressure. Airflow was measured by connecting the proximal end of the endotracheal tube to a pneumotachograph.
  • the pressure transducer-catheter system was dynamically balanced, and no phase shift was detectable between pressure and flow up to a frequency of 9 Hz.
  • the signals of transpulmonary pressure and flow were recorded on a multichannel physiological recorder which was linked to a personal computer for on-line calculation of mean pulmonary flow resistance (R L ) by dividing the change in transpulmonary pressure by the change in flow at mid- tidal volume (obtained by digital integration).
  • Aerosols are generated using a disposable medical nebulizer that provided an aerosol with a mass median aerodynamic diameter of 3.2 ⁇ m as determined by a cascade impactor.
  • the nebulizer was connected to a dosimeter system, consisting of a solenoid valve and a source of compressed air (20 psi).
  • the output of the nebulizer was directed into a plastic T-piece, one end of which was connected to the inspiratory port of a respirator.
  • the solenoid valve was activated for one second at the beginning of the inspiratory cycle of the respirator. Aerosols were delivered at a tidal volume of 500 ml and a rate of 20 breaths per minute.
  • COMPOUND 2 was
  • Fully conditioned mongrel dogs will be anesthetized with an intravenous lidocaine drip 1 mg/min and maintained throughout the occlusion and reperfusion.
  • a bolus dose of Procainimide (10mg/kg/hr) will be infused prior to the occlusion and an intravenous infusion of 3 mg/kg/hr will be maintained throughout the procedure.
  • the left anterior descending artery will be occluded using the snare and the occlusion will be maintained for two hours. Flow cessation will be documented by the doppler flow signal. Hemodynamic, functional, and microsphere blood flow measurements will be repeated at ten minutes into the occlusion, one hour post-occlusion, and just before treatment with COMPOUND 2 (one hour 45 minutes post- occlusion). The placebo or TBC compound will be infused over ten minutes using a Harvard infusion pump.
  • Hemodynamic and functional measurements will be repeated during the treatment (1hr 50min post-occlusion), and two hours post-occlusion. At two hours post-occlusion the occluder will be released and re-flow will be reestablished and documented using the doppler flow probe. Ten minutes after reperfusion, hemodynamic, functional, and microsphere blood flow measurements will be repeated. Hemodynamic and functional measurements will be repeated at one and two hours post-reperfusion.
  • the animal will be sacrificed using super saturated potassium chloride according to animal welfare guidelines.
  • the heart will be quickly excised from the chest and washed with tap water.
  • a perfusion cannula will be inserted in to the left anterior descending coronary at the level of the occluder and the ascending aorta will be attached to a perfusion stand.
  • the left anterior descending coronary will be perfused with triphenyl tetrazolium chloride, (1% solution) and the aortic root will be perfused with Evans Blue at equal pressure (100 mm Hg) for four minutes.
  • the heart After perfusion, the heart will be taken down from the perfusion stand, the atria and right ventricle will be excised, and the left ventricle will be sliced in one cm thick sections, in breadloaf fashion from base to apex and weighed.
  • TTC will stain the risk region, which has residual viable tissue, brick red, while the infarcted tissue will remain nonstained and appear tan.
  • the control region will appear blue, having been stained by the Evans Blue.
  • Tissue samples from the infarct zone, the border zone, risk region and the non-ischemic or control region will be obtained and flash frozen at -70°C in liquid nitrogen for a myeloperoxidase assay to quantitate the relative presence of neutrophil infiltration.
  • the slices will be incubated in TTC at 37°C for 30 additional minutes in insure proper staining of ischemic, but viable, tissue.
  • TTC staining the slices will be weighed again and traced on acetate film, both basal and apical views. These respective areas will be planimetered to determine the percent area of each slice occupied by infarct, risk region or control region. These percent areas will be multiplied by individual slice weights to determine the actual infarct weight as well as the infarct percent of the risk region weight, and of the total left ventricular weight.
  • the slices will be fixed in formalin, photographed, and samples will be taken from representative endocardial, midwall, and epicardial areas for calculation of
  • MPO myeloperoxidase
  • CBV'S in coronary blood flow: Thirty five mongrel dogs of either sex weighing between 21-32 kg were used. Dogs were sedated with 5mg I.M. Acepromazine, anesthetized with 30mg/kg I.V.
  • Pentobarbital sodium and ventilated with a mechanical respirator (Harvard model 613, South Natick, MA).
  • Plastic catheters were placed in a carotid artery for aortic pressure measurements and in a jugular vein for fluid and drug administration.
  • a left fifth intercostal thoracotomy was performed and the heart suspended in a pericardial cradle.
  • An approximately 2cm segment of the LAD coronary artery was carefully isolated and the nearby branches ligated.
  • An ultrasonic Doppler flow probe (Hartley instruments, Houston, TX) was placed around the proximal portion of the isolated segment of the LAD to measure phasic and mean coronary blood flow velocities.
  • Baseline hemodynamics including heart rate, systolic and diastolic blood pressures, phasic and mean coronary blood flow velocities, were recorded on a multichannel recorder (Gould Inc. model 3800, Cleveland, OH).
  • CFV's were induced in the isolated LAD segment by first gently squeezing the vessel with a cushioned forceps to damage the endothelium, followed by the placement of an extermal plastic constrictor over the damaged area to reduce blood flow velocity. After endothelial injury and extermal constriction of the LAD coronary artery, cyclic flow variations, or CFV's
  • COMPOUND 2 was given intravenously as a 50mg/kg bolus followed by an infusion started at 50mg/kg/hr rate, with increments every 30 minutes to 100, 200, and 400mg/kg/hr maximum rate.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Ce procédé qui permet d'inhiber la liaison de E-sélectine, P-sélectine et/ou L-sélectine aux sialyl-Lewisx, sialyl-Lewisa, Lewis x et/ou Lewisa présent à la surface d'une cellule, consiste à administrer à un patient nécessitant un tel traitement une dose pharmacologiquement efficace d'au moins un composé de formule XCOCH¿2?CH2COOR3 dans laquelle X représente (a); (b); (c); (d); ou (e). R1 représente un groupe alkyle ou alcoxy ou un atome d'halogène; R2 représente un atome d'halogène ou d'hydrogène; et R3 représente un groupe alkyle ou un atome d'hydrogène; (I) où R1 a la notation ci-dessus définie et AA représente un acide aminé α; (II); (III) où R3 représente NO2; (f); on décrit ce composé ainsi que ses sels, esters, amides et promédicaments pharmacologiquement acceptables.
PCT/US1996/006453 1995-05-08 1996-05-06 Procede d'inhibition de la liaison de selectines aux sialyl-lewis WO1996035418A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU57322/96A AU5732296A (en) 1995-05-08 1996-05-06 Method for inhibiting the binding of selectins to sialyl-lew ises

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US43612695A 1995-05-08 1995-05-08
US08/436,126 1995-05-08

Publications (1)

Publication Number Publication Date
WO1996035418A1 true WO1996035418A1 (fr) 1996-11-14

Family

ID=23731211

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/006453 WO1996035418A1 (fr) 1995-05-08 1996-05-06 Procede d'inhibition de la liaison de selectines aux sialyl-lewis

Country Status (2)

Country Link
AU (1) AU5732296A (fr)
WO (1) WO1996035418A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6235309B1 (en) 1997-02-28 2001-05-22 The Regents Of The University Of California Inhibition of cell-cell binding by lipid assemblies
JP2007538014A (ja) * 2004-05-14 2007-12-27 エミスフェアー・テクノロジーズ・インク 活性薬剤を送達するためのアリールケトン化合物および組成物
EP3494966A1 (fr) 2013-03-13 2019-06-12 Cour Pharmaceuticals Development Company Particules de modification immunitaire pour le traitement d'inflammation
WO2019217780A1 (fr) 2018-05-11 2019-11-14 Phosphorex, Inc. Microparticules et nanoparticules ayant des charges de surface négatives
EP3569226A1 (fr) 2012-12-04 2019-11-20 Phosphorex, Inc. Microparticules et nanoparticules dotées de charges de surface négatives

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3904682A (en) * 1967-01-13 1975-09-09 Syntex Corp 2-(6{40 -Methoxy-2{40 -naphthyl)acetic acid
US4009197A (en) * 1967-01-13 1977-02-22 Syntex Corporation 2-(6-Substituted-2'-naphthyl) acetic acid derivatives and the salts and esters thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3904682A (en) * 1967-01-13 1975-09-09 Syntex Corp 2-(6{40 -Methoxy-2{40 -naphthyl)acetic acid
US4009197A (en) * 1967-01-13 1977-02-22 Syntex Corporation 2-(6-Substituted-2'-naphthyl) acetic acid derivatives and the salts and esters thereof

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6235309B1 (en) 1997-02-28 2001-05-22 The Regents Of The University Of California Inhibition of cell-cell binding by lipid assemblies
US6663886B2 (en) 1997-02-28 2003-12-16 Regents Of The University Of California Inhibition of cell-cell binding by lipid assemblies
JP2007538014A (ja) * 2004-05-14 2007-12-27 エミスフェアー・テクノロジーズ・インク 活性薬剤を送達するためのアリールケトン化合物および組成物
US9035085B2 (en) 2004-05-14 2015-05-19 Emisphere Technologies, Inc. Aryl ketone compounds and compositions for delivering active agents
EP3569226A1 (fr) 2012-12-04 2019-11-20 Phosphorex, Inc. Microparticules et nanoparticules dotées de charges de surface négatives
US11771654B2 (en) 2012-12-04 2023-10-03 Cytodigm, Inc. Microparticles and nanoparticles having negative surface charges
EP3494966A1 (fr) 2013-03-13 2019-06-12 Cour Pharmaceuticals Development Company Particules de modification immunitaire pour le traitement d'inflammation
WO2019217780A1 (fr) 2018-05-11 2019-11-14 Phosphorex, Inc. Microparticules et nanoparticules ayant des charges de surface négatives

Also Published As

Publication number Publication date
AU5732296A (en) 1996-11-29

Similar Documents

Publication Publication Date Title
Hunt et al. Endotoxemia, disturbance of coagulation, and obstructive jaundice
EP0089938B1 (fr) Compositions à emploi thérapeutique ou diagnostique contenant des oligosaccharides
AU733692B2 (en) Inhibition of cell-cell binding by lipid assemblies
Cundell et al. Relationship between colonial morphology and adherence of Streptococcus pneumoniae
Asako et al. Indomethacin-induced leukocyte adhesion in mesenteric venules: role of lipoxygenase products
US5654282A (en) Selectin binding glycopeptides
US20080112955A1 (en) Method and composition for preventing pain in sickle cell patients
US7365080B2 (en) Pharmaceutical compositions comprising thieno[2,3-c]pyridine derivatives and use thereof
CA2182007A1 (fr) Derives de lactose substitues, utilises comme inhibiteurs de l'adhesion cellulaire
EP0369463B1 (fr) Conjugué de prostaglandine et polysaccharide
Rachmilewitz et al. Prostanoid synthesis by cultured peripheral blood mononuclear cells in inflammatory diseases of the bowel
WO1996035418A1 (fr) Procede d'inhibition de la liaison de selectines aux sialyl-lewis
CA2044328A1 (fr) Efomycines a.e et g utilisees comme agents anti-inflammatoires
WO2005089068A2 (fr) Compositions pharmaceutiques comprenant des derives de quinazolinecarboxamide anti-inflammatoires
EP0380084A2 (fr) Méthode d'inhibition des interactions cellule-cellule et cellule-substrat par blocage des interactions entre hydrates de carbone
Gionchetti et al. Bronchopulmonary involvement in ulcerative colitis
Bodenheimer Jr et al. Characterization of a new monoclonal antibody to rat macrophages and Kupffer cells
US20070191400A1 (en) Pharmaceutical compositions comprising anti-inflammatory quinazolinecarboxamide derivatives
JPH05504550A (ja) 免疫抑制剤として有用なピリミジン生合成インヒビター
JP3769045B2 (ja) 抗炎症剤
Fujimoto et al. Effect of the 21-aminosteroid U-74006F on antigen-induced bronchoconstriction and bronchoalveolar eosinophilia in allergic sheep
Zopf et al. Development of anti-adhesion carbohydrate drugs for clinical use
JPH05105635A (ja) 抗ウイルス剤
RU2117482C1 (ru) Средство антиастматического и антиадгезионного действия
JP4143169B2 (ja) ノジリマイシン亜硫酸付加物のアシル化物

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU BR CA CN CZ FI HU JP KR MX NO NZ PL RU UA US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA