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
  • A61K31/13—Amines
  • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
• • 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
• • 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
  • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
  • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
  • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P13/00—Drugs for disorders of the urinary system
  • A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
  • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• NRL-lpr/lpr mice have been studied as a model for human autoimmune diseases. This strain of mice develops a spontaneous autoimmune disease characterized by lymphadenopathy, autoantibody production and inflammatory manifestations including nephritis, vasculitis, and arthritis (Hang, L. , et al . , J. Exp. Med . 155:1690-1701 (1982); Eisenberg, R.A., et al . , Clin . Exp. heumatol . 7:S35-S40 (1989)).
• Immune function abnormalities also include enhanced constitutive macrophage class II antigen expression, elevated levels of IFN- ⁇ , TNF, IL-1, and IL-6 in isolated kidney, lymph node, and spleen cells, and an enhanced state of macrophage "activation".
• These disease manifestations are a result of both a single gene mutation (lpr) of the Fas apoptosis gene on mouse Chromosome 19 and background genes from the MRL strain.
• the MRL genes contributing to disease manifestations have not been identified, two loci contributing to renal disease have been mapped to regions of mouse Chromosomes 7 and 12.
• NO nitric oxide
• NOS nitric oxide synthase
• NO also has potent proinflam atory actions. NO may increase vascular permeability in inflamed tissues. Pain is also an important aspect of inflammation. Several groups have now demonstrated that NO plays a role in the mediation of pain in inflammation. Intradermal injection of solutions containing NO into humans caused a dose-related occurrence of pain in the site (Holthusen, H. , and Arndt, J.O., Neuroscience Letters 165:71-74 (1994) . Thus, pain, a hallmark symptom of inflammation, can be induced by NO.
• NO has also been shown to cause increased production of TNF and IL-1 by cells, and to increase the potential of cells to produce hydrogen peroxide.
• rabbit and human chondrocytes have been shown to produce NO and to express inducible nitric oxide synthase (iNOS) in response to various cytokines and bacterial products.
• NO is an important mediator in immune complex vasculitis in rats (Mulligan, M.S., et al . , Brit . J. Pharmacol . 107:1159-1162 (1992)).
• Some researchers have noted a role for NO in inflammatory bowel disease in guinea pigs (Miller, M.J., et al . , J. Pharmacol . Exp. Ther.
• Nitrotyrosine is formed by the action of peroxynitrite on tyrosine, and it is felt to be a stable "footprint” or “track” of the presence of NO (Beckman, J.S. et al . , Methods Enzymol . 233:229-240 (1994)). Nitrated proteins have been found associated with macrophages and inflammation (by using an anti-nitrotyrosine antibody) in atheromatous plaques in human vessels (Beckman, J.S., et al., Biol . Chem . Hoppe- Seyler 375:81-88 (1994)), providing further evidence that nitrotyrosine is formed in vivo in humans.
• the invention relates to a method of treating or preventing autoimmune diseases, such as rheumatoid arthritis, insulin dependent diabetes mellitus, systemic lupus erythematosus and glomerulonephritis, comprising administering, preferably enterally, to a patient in need thereof an effective amount of a nitric oxide synthase inhibitor or a nitric oxide scavenger.
• autoimmune diseases such as rheumatoid arthritis, insulin dependent diabetes mellitus, systemic lupus erythematosus and glomerulonephritis
• the Figure is a bar graph showing the scores of pathological characteristics in MKL-lpr/lpr mice either treated with N G -monomethyl-L-arginine (NMMA) or left untreated.
• nitric oxide has now been linked to the manifestation of autoimmune diseases, particularly chronic diseases, such as rheumatoid arthritis, insulin dependent diabetes mellitus, systemic lupus erythematosus and glomerulonephritis.
• autoimmune diseases particularly chronic diseases, such as rheumatoid arthritis, insulin dependent diabetes mellitus, systemic lupus erythematosus and glomerulonephritis.
• the inhibitors of nitric oxide synthase known to synthesize nitric oxide in vivo, or nitric oxide scavengers can be useful in the prevention or treatment of autoimmune diseases (Gilkeson et al., Arth. Rheum. 36 (Suppl.) :S219, 1993 (September)).
• Inhibitors of nitric oxide synthase which can be used in this invention are those known in the art and include substrate analogs, such as aminoguanidine, N°-amino-L- arginine, N G -methyl-L-arginine, N G -nitro-L-arginine, N G - nitro-L-arginine methyl ester, and N G -iminoethyl-L- ornithine, flavoprotein binders, such as diphenylene iodonium, iodonium diphenyl and di-2-thienyl iodonium, calmodulin binders, such as calcineurin, trifluoroperazine, N-(4-aminobutyl)-5-chloro-2- naphthalenesulfonamide and N-(6-aminohexyl)-1- naphthalenesulfonamide, heme binders, such as carbon monoxide, deple
• N G -amino-L-arginine N G -methyl-L-arginine, N°-nitro-L-arginine, N°-nitro-L-arginine methyl ester, and N°-iminoethyl-L-ornithine.
• N G - amino-L-arginine N°-methyl-L-arginine, N G -nitro-L-arginine, and aminoguanidine.
• N°- methyl-L-arginine Pharmaceutically acceptable salts may also be administered.
• Suitable salts include acid salts, such as hydrogen chloride, hydrogen bromide, hydrogen iodide, sulfate and acetate salts, as well as basic salts, such as a ine, ammonium, alkali metal and alkaline earth metal salts.
• Scavengers of nitric oxide are compounds which will react with nitric oxide in vivo, such as hemoglobin (Wang et al . , Life Sciences 49:55-60 (1991)) and cobalamins (Rajanayagam, C.G., et al . , Brit . J. Pharmacol . 108:3-5 (1993); Zatarain, J. , et al . , Clin . Res . 41:783A (1993)).
• the compounds of the claimed invention can be administered alone or in a suitable pharmaceutical composition. Modes of administration are those known in the art, such as enteral, parenteral or topical application. Enteral is preferred and oral administration is particularly preferred.
• Suitable pharmaceutical carriers can be employed and include, but are not limited to water, salt solutions, alcohols, polyethylene glycols, gelatin, carbohydrates such as lactose, amylose or starch, magnesium stearate, talc, silicic acid, viscous paraffin, fatty acid esters, hydroxymethylcellulose, polyvinyl pyrrolidone, etc.
• the pharmaceutical preparations can be sterilized and if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, flavorants, coloring, and/or aromatic substances and the like which do not deleteriously react with the active compounds.
• auxiliary agents e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, flavorants, coloring, and/or aromatic substances and the like which do not deleteriously react with the active compounds.
• injectable, sterile solutions preferably oily or aqueous solutions, as well as suspensions, emulsions, or implants. including suppositories.
• Ampoules are convenient unit dosages.
• Oral applications are preferably administered in the forms of capsules, tablets and/or liquid formulations. Unit form dosages are preferred.
• Topical applications can be administered in the form of a liquid, gel or a cream.
• the actual amounts of the active compounds in a specific case will vary according to the specific compound being utilized, the particular composition formulated, the mode of administration and the age, weight and condition of the patient, for example.
• Dosages for a particular patient can be determined by one of ordinary skill in the art using conventional considerations, (e.g. by means of an appropriate, conventional pharmacological protocol) .
• the invention is further specifically illustrated by the following exemplification.
• mice were housed in metabolic cages (3 per cage) and fed deionized-distilled sterile water and a defined arginine and nitrate-free diet. Urine was collected into isopropanol to inhibit bacterial growth. Urinary nitrite/nitrate concentration was determined spectrophotometrically as described before (Granger, D.L. et al . , J. Immunol . 146:1294 (1991)). Determinations were done in duplicate or triplicate. Urinary protein was measured by the Bradford assay. Total nitrate excretion was then calculated based on the concentration and the urine volume.
• nitrite/nitrate In animals consuming a nitrate-free diet, urinary excretion of nitrite/nitrate accurately reflects the endogenous production of NO.
• urine collected daily under basal conditions MRL-lpr/lpr mice excrete more nitrite/nitrate than do C3H mice, when analyzed over a 10 day period at age 3 months.
• MRL- lpr/lpr mice excrete higher levels of nitrite/nitrate than do B6 mice as the mice age. Higher nitrate/nitrite excretion begins at approximately age 10 to 12 weeks paralleling that of proteinuria.
• Spleen, liver, kidneys, lymph nodes, and peritoneal cells were collected and quickly frozen in a dry ice-ethanol slurry in a buffer-protease inhibitor cocktail containing 100 ⁇ M phenylmethylsulfonyl fluoride, 5 ⁇ g/ l aprotinin, 1 ⁇ g/ml chymostatin, and 5 ⁇ g/ml pepstatin A.
• the tissue cells were then disrupted with a pestle in repeated freeze-thaw cycles. Cytosol was collected after centrifugation, and assayed for protein and NOS activity using a modification of procedures known previously (Bredt, D.S. and Snyder, S.H. , Proc . Natl . Acad . Sci.
• the assay buffer contained 50 mM HEPES (pH 7.5), 200 ⁇ M NADPH, 1 mM dithiothreitol, 10 ⁇ M FAD, 100 ⁇ M tetrahydrobiopterin, and 10 ⁇ M L-arginine.
• HEPES pH 7.5
• NADPH 1 mM dithiothreitol
• 10 ⁇ M FAD 100 ⁇ M tetrahydrobiopterin
• L-arginine L-arginine labeled with tritium in the guanidino position. Thirty microliters of sample were used in a total reaction mixture of 50 microliters. Samples were done in duplicate or triplicate.
• Peritoneal macrophages from the normal BALB mice had no enhancement of nitrite/nitrate production when treated with endotoxin or IFN-7 alone, but the combination enhanced the production greatly.
• peritoneal macrophages from MRL-lpr/lpr mice had enhanced responses to treatment with endotoxin and murine IFN-7 alone, as well as with combined endotoxin IFN- ⁇ treatment.
• tissue iNOS mRNA expression was extracted from organs from BALB and MRL-lpr/lpr mice, and then examined by Northern analysis for iNOS mRNA expression.
• the iNOS mRNA (approximately 4.7 kilobases in size) was noted in tissue from kidney and spleen from MRL-lpr/lpr mice, but not in those tissues from BALB mice.
• Various tissues and cells from MRL-lpr/lpr and BALB mice were extracted and analyzed for their abilities to convert 14 C-L-arginine (labeled in the guanidino position) to 14 C-L-citrulline.
• Peritoneal macrophages and spleen extracts from MRL-lpr/lpr mice displayed more NOS activity than did those from BALB mice, while the activity in kidney extracts was not different.
• tissue extracts were analyzed for iNOS antigen by immunoprecipitation and immunoblotting techniques using a rabbit anti-mouse iNOS antibody, we did not detect antigen in extracts from organs of BALB mice, although extracts from spleen and kidney tissues from MRL-lpr/lpr mice had readily detectable antigen.
• mice Groups of 8 week old MRL-lpr/lpr mice were given either sterile, distilled deionized water or water containing 50 mM NMMA for ad libitum consumption.
• NMMA was from CalBiochem (San Diego, CA) and from Dr. Owen Griffith (Milwaukee, WI) . Both groups of mice were maintained on the defined nitrate free diet described above. At weekly intervals, the mice were placed in metabolic cages and 24 hour urine collections were obtained. Urinary nitrite/nitrate was measured as described above, and urinary protein was determined using the Bradford assay (BioRad, Hercules, CA) . After 10 weeks of treatment, the mice were bled and sacrificed with removal of the kidneys and knee joints.
• Serum anti-DNA activity was determined by ELISA as previously described.
• the kidneys were imbedded in paraffin, sectioned and stained with hematoxylin and eosin. Knee joints were decalcified in folic acid, embedded into paraffin, sectioned, and stained. Slides were then read by a pathologist "blinded” as to the group of origin. The amount of kidney and knee joint disease present in each specimen was quantitated as noted before. Briefly, glomeruli were graded for hypercellularity (0-4) , hyperlobularity (0-4) , crescents (0-4) , and necrosis (0-4) . A score was then derived by adding the grading of these features of glomerular disease.
• Kidneys from normal BALB mice usually have scores from 0-1. Vasculitis was noted when present in medium size vessels in the kidney sections. The synovial score was derived by adding the grading of synovial proliferation (0-3) and subsynovial inflammation (0-3) . Knee joints from normal BALB mice usually have scores from 0-0.5.
• NMMA in the drinking water of MRL-lpr/lpr mice effectively blocked nitrite/nitrate excretion (and by inference nitric oxide production) . Also, mice receiving NMMA excreted significantly less protein than did control mice; this difference became apparent at week five of treatment.
• the chronic interstitial lymphocytic infiltrate seen in the kidneys of all lpr congenic mice was present to comparable degrees in both control and NMMA treated mice.
• NO-Hb Formation of nitroso-hemocrlobin NO-Hb forms through an interaction of NO with iron in the heme group of hemoglobin (Huot, A.E., et al . , Biochem . Biophys . Res . Commun . 182 : 151-157 , (1992); Cantilena, L.R.J., et al . , J. Lab . Clin . Med . 120:902-907 (1992)).
• Whole blood from MRL-lpr/lpr mice at different ages was analyzed for the presence of nitroso-hemoglobin (NO-Hb) .
• the blood samples were anticoagulated and examined by electron paramagnetic resonance (EPR) at 77°K using a Bruker ESP300 spectrometer (Cha ulitrat, W. et al . , Molec . Pharmacol . 46:391-397 (1994)).
• An age-dependent increase was observed in the amount of NO-Hb in the blood of the diseased mice.
• the levels of NO-Hb were higher in MRL-lpr/lpr mice compared to same-age control mice without disease. The differences were statistically significant (p ⁇ 0.05 at all ages analyzed).
• the presence of NO-Hb is another important sign that NO is being over-expressed in these mice with autoimmune nephritis and arthritis.
• NO may react with superoxide and form the highly reactive, tissue destructive molecule peroxynitrite.
• cells from MRL-lpr/lpr mice can overproduce reactive oxygen species such as hydrogen peroxide, superoxide (Dang-Vu, A.P. et al . , J. Immunol . 138:1757-1761 (1987)) and nitric oxide (Weinberg, J.B., et al . , J. Exp. Med . 179:651-660 (1994)).
• a study was done to look for evidence that MRL- lpr/lpr mice also overproduce the destructive molecule peroxynitrite (Beckman, J.S., et al .
• Kidney tissue was homogenized with a glass pestle. Proteins in soluble extracts (100 ⁇ g per lane) were separated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose (0.45 ⁇ m, Novex) . Unbound sites were blocked by incubation with 1% non-fat dry milk in TBS (20 mM Tris, 500 M NaCl, pH 7.5) for 60 min at 25°C. Membranes were incubated overnight at 25°C with a polyclonal anti-nitrotyrosine antibody (0.25 ⁇ g/ml) in 1% milk/TBS.
• Catalase activity in kidneys of MRL-lpr/lpr mice The catalase content in kidneys from 20 week old normal (BALB/c) and MRL-lpr/lpr mice was analyzed. Peroxynitrite or NO can destroy catalase activity. Catalase was measured by the disappearance of hydrogen peroxide noted by absorbance at 240 nm (Beers, R.F., and Sizer, R.W. , J. Biol . Chem . 195:133 (1952)). Values shown in Table 2 are the mean ⁇ SEM of replicate samples expressed in units/mg protein. Table 2 shows that kidneys from the control mice had high levels of catalase while levels of catalase from MRL-lpr/lpr mice were very low.
• H 2 0 2 was added to a final concentration of 5.0 mM and the gel was incubated an additional 15 min. After a brief rinse in water, stain development was initiated by addition of 0.5 mg/ml diaminobenzidine-HCl in 50 mM sodium phosphate, 0.1 mM EDTA, pH 7.0. Results showed clearly that kidneys from 4 different BALB/c control mice contained large amounts of catalase, while those from two MRL-lpr/lpr mice had markedly diminished levels of catalase.
• iNOS protein expressed in their synovial tissues were studied by immunofluorescence techniques using mouse monoclonal anti-iNOS antibody (purchased from Transduction Laboratories, Inc.).
• mouse monoclonal anti-iNOS antibody purchased from Transduction Laboratories, Inc.
• iNOS antigen was detected in the tissues of two of three rheumatoid arthritis patients and in one of three osteoarthritis patients.

Landscapes

• Health & Medical Sciences (AREA)
• Medicinal Chemistry (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• General Health & Medical Sciences (AREA)
• Animal Behavior & Ethology (AREA)
• Life Sciences & Earth Sciences (AREA)
• Pharmacology & Pharmacy (AREA)
• Chemical & Material Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Organic Chemistry (AREA)
• General Chemical & Material Sciences (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Engineering & Computer Science (AREA)
• Diabetes (AREA)
• Epidemiology (AREA)
• Endocrinology (AREA)
• Emergency Medicine (AREA)
• Hematology (AREA)
• Obesity (AREA)
• Immunology (AREA)
• Pain & Pain Management (AREA)
• Rheumatology (AREA)
• Urology & Nephrology (AREA)
• Reproductive Health (AREA)
• Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Applications Claiming Priority (3)

Publications (1)

Family

ID=22547353

Family Applications (1)

Country Status (5)

Families Citing this family (16)

Family Cites Families (3)

• 1994
  • 1994-11-17 JP JP7514592A patent/JPH09508891A/ja active Pending
  • 1994-11-17 CA CA002176747A patent/CA2176747A1/en not_active Abandoned
  • 1994-11-17 AU AU12099/95A patent/AU1209995A/en not_active Abandoned
  • 1994-11-17 WO PCT/US1994/013239 patent/WO1995013805A1/en not_active Application Discontinuation
  • 1994-11-17 EP EP95903124A patent/EP0729356A1/de not_active Withdrawn

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events