WO2009003970A1 - Ciblage de promédicaments diazo pour le traitement de maladies gastro-intestinales - Google Patents

Ciblage de promédicaments diazo pour le traitement de maladies gastro-intestinales Download PDF

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WO2009003970A1
WO2009003970A1 PCT/EP2008/058338 EP2008058338W WO2009003970A1 WO 2009003970 A1 WO2009003970 A1 WO 2009003970A1 EP 2008058338 W EP2008058338 W EP 2008058338W WO 2009003970 A1 WO2009003970 A1 WO 2009003970A1
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alkyl
compound
hydrogen
group
drug
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PCT/EP2008/058338
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John Francis Gilmer
Juan Francisco Marquez
Dermot Kelleher
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The Provost, Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth, Near Dublin
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Priority to AU2008270363A priority Critical patent/AU2008270363A1/en
Priority to EP08774496A priority patent/EP2160377A1/fr
Priority to CN200880022191A priority patent/CN101801919A/zh
Priority to JP2010513951A priority patent/JP2010531845A/ja
Priority to CA002691716A priority patent/CA2691716A1/fr
Publication of WO2009003970A1 publication Critical patent/WO2009003970A1/fr

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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C245/00—Compounds containing chains of at least two nitrogen atoms with at least one nitrogen-to-nitrogen multiple bond
    • C07C245/02—Azo compounds, i.e. compounds having the free valencies of —N=N— groups attached to different atoms, e.g. diazohydroxides
    • C07C245/06—Azo compounds, i.e. compounds having the free valencies of —N=N— groups attached to different atoms, e.g. diazohydroxides with nitrogen atoms of azo groups bound to carbon atoms of six-membered aromatic rings
    • C07C245/08—Azo compounds, i.e. compounds having the free valencies of —N=N— groups attached to different atoms, e.g. diazohydroxides with nitrogen atoms of azo groups bound to carbon atoms of six-membered aromatic rings with the two nitrogen atoms of azo groups bound to carbon atoms of six-membered aromatic rings, e.g. azobenzene
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/12—Antidiarrhoeals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • 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
    • A61P33/00—Antiparasitic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
    • A61P33/04—Amoebicides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06—Esters of carbamic acids
    • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
    • C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C271/20—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by nitrogen atoms not being part of nitro or nitroso groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C309/00—Sulfonic acids; Halides, esters, or anhydrides thereof
    • C07C309/01—Sulfonic acids
    • C07C309/28—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
    • C07C309/45—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing nitrogen atoms, not being part of nitro or nitroso groups, bound to the carbon skeleton
    • C07C309/46—Sulfonic acids having sulfo groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton containing nitrogen atoms, not being part of nitro or nitroso groups, bound to the carbon skeleton having the sulfo groups bound to carbon atoms of non-condensed six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C309/00—Sulfonic acids; Halides, esters, or anhydrides thereof
    • C07C309/63—Esters of sulfonic acids
    • C07C309/64—Esters of sulfonic acids having sulfur atoms of esterified sulfo groups bound to acyclic carbon atoms
    • C07C309/69—Esters of sulfonic acids having sulfur atoms of esterified sulfo groups bound to acyclic carbon atoms of a carbon skeleton substituted by nitrogen atoms, not being part of nitro or nitroso groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D319/00—Heterocyclic compounds containing six-membered rings having two oxygen atoms as the only ring hetero atoms
    • C07D319/04—1,3-Dioxanes; Hydrogenated 1,3-dioxanes
    • C07D319/08—1,3-Dioxanes; Hydrogenated 1,3-dioxanes condensed with carbocyclic rings or ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
    • C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
    • C07D407/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
    • C07F9/40—Esters thereof
    • C07F9/4003—Esters thereof the acid moiety containing a substituent or a structure which is considered as characteristic
    • C07F9/4056—Esters of arylalkanephosphonic acids
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    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
    • C07F9/40—Esters thereof
    • C07F9/4071—Esters thereof the ester moiety containing a substituent or a structure which is considered as characteristic
    • C07F9/4075—Esters with hydroxyalkyl compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2603/00—Systems containing at least three condensed rings
    • C07C2603/02—Ortho- or ortho- and peri-condensed systems
    • C07C2603/40—Ortho- or ortho- and peri-condensed systems containing four condensed rings

Definitions

  • the present application relates to compounds and pharmaceutical compositions that may be used for the treatment of gastrointestinal diseases, including, but not limited to inflammatory diseases, such as Inflammatory Bowel Disease (IBD). BACKGROUND OF THE INVENTION
  • Drug targeting may be defined as the delivery of a drug to a specific organ, tissue or cell population (Schreier 2001). Still in its infancy, this field offers the prospect of enhancing the efficacy of drug treatment while reducing systemic impact or side effects. Despite the promise of this approach, and ongoing efforts, there have been few successful examples to date due in part to limited understanding of the basic factors underlying drug transport and the expression of potential targeting vectors. Chemical drug targeting involves the deliberate modification of a drug structure (usually bioreversibly) causing it to accumulate in a target tissue; site-specific release from the prodrug is triggered by a chemical or enzymatic condition not present elsewhere in the body.
  • the colon is an important challenge to the validity of the drug targeting approach, as conditions in the colon are largely similar to those prevailing elsewhere in the gastrointestinal (GI) system, and the luminal pH gradient through the GI tract is too gradual for effective local drug release on strictly chemical grounds (Bauer, 2001).
  • the colon is an important drug target for the treatment of pathologies of the colon itself, such as inflammatory bowel disease (IBD) and colon cancer, and for the relief of the chronic constipation that accompanies opioid drug treatment.
  • IBD inflammatory bowel disease
  • the colon is also important as a potential portal site for peptide and protein drugs that are not absorbed from other regions of the GI tract or are too unstable in the presence of duodenal proteases to be released there (Saffian, 1986; Bai, 1995).
  • the human GI system is home to 400-500 species of bacteria with a total live population of 10 14 organisms. This is remarkable when compared with the 10 13 eukaryotic cells that make up the human body.
  • the GI tract has a steadily increasing bacterial concentration gradient on descending from the stomach through the small intestine, followed by an enormous increase at the colon.
  • the bacterial concentration in the small intestine is typically 10 3 -10 4 CFU mf 1 whereas the concentration in the colon is 10 ⁇ -10 12 CFU mf 1 and one third of fecal dry weight consists of bacteria (Moore and Holdeman, 1974 and 1975; Simon and Gorbach, 1984).
  • These organisms fulfill their energy needs by fermenting undigested materials entering from the small intestine (particularly polysaccharides) and have for this purpose evolved an elaborate array of enzymes such as azoreductase, glucosidase, ⁇ -glucuronidase, ⁇ -xylosidase, nitroreductase, galactosidase and deaminase (Scheline, 1973).
  • the drug is selected from the group that includes an anti-inflammatory drug, an anti-cancer drug, an imaging agent, particularly as used in the imaging of colon diseases; a vaccine, an antigen, an anti-infective drug, a peptide, an antisense molecule and a protein.
  • the method provides an evaluation of antiinflammatory effects of candidate compounds in a mouse model of IBD.
  • the method provides insights into the GI permeability of the azo-compounds and their ability to pass through the GI tract.
  • a method comprising administering a therapeutically effective amount of a compound or composition effective to reduce, alleviate or treat various gastrointestinal diseases, including inflammatory bowel disease (IBD).
  • IBD inflammatory bowel disease
  • the drug is linked via an ester group to a carrier group, which is connected by an azo-bond to a second carrier group.
  • the carrier groups may be directly attached to the azo group or indirectly attached to the azo group.
  • the carrier groups are designed to maximally suppress absorption from stomach and upper intestine.
  • the method exploits the selective reduction of an azo-linker in the colon, releasing a chemically unstable, latent prodrug that subsequently undergoes cyclization, such as lactamization, that liberates the drug payload, such as a steroid.
  • the cyclization reaction is substantially spontaneous.
  • the prodrugs of the present application are referred to as “carrier-drug.”
  • the “carrier” can comprise compounds such as 5-ASA or para-aminobenzoic acid (PABA).
  • the carrier has a therapeutic effect, such as 5-ASA
  • the prodrug is generally referred to as a "mutual prodrug.”
  • Such mutual prodrugs can be referred to herein as 5-ASA-drug, wherein the drug can be any appropriate therapeutic agent, including those disclosed herein.
  • Such mutual prodrugs can include, but are not limited to, 5-ASA-ciprofloxacin, 5-ASA-bevacizumab, 5-ASA-prednisolone, 5-ASA-5-ASA, etc.
  • the carrier does not have a therapeutic effect, such as PABA
  • the compound can be simply referred to as a "prodrug.”
  • Such prodrugs can be referred to herein as PABA-drug, such as, for example PABA-ciprofloxacin, PABA-bevacizumab, PAB A-predniso lone,
  • Azo bond reduction proceeds readily because it is based on the promiscuity of the azoreductases present in the colon with respect to substrate, as evidenced by their ability to efficiently reduce substrates as diverse as ipsalazide, in which the carrier group is /?-amino hippurate, balsalzide (p-aminobenzoyl- ⁇ -alanine carrier), sulfasalazine (sulfapyridine carrier), sterically bulky PAF antagonists (Carceller et al., 2001), 9-aminocamphothecin (Sakuma et al., 2001) and 5-ASA-N-methacrylamide, acryloyloxyethyl and acryloylamido copolymers (e.g. Van den Mooter et al., 1994).
  • the presence of the vast microflora in the bowel causes a change in redox potential from -67 ⁇ 90 in the distal small bowel to -
  • ester linkage in the compounds of the present application is highly stable under conditions found in the GI tract. Esterase activity in the lumen of the GI tract is restricted to the pancreatic serine proteases, which exhibit residual esterase activity towards a limited number of substrates, generally esters of aromatic amino acids. Steroidal 21 -esters (see Scheme 2 above for numbering), for example, have been shown to be robust in simulated intestinal fluid models. For example Fleisher et al. (1986) reported a first-order rate constant of 0.003 min "1 for the hydrolysis of hydrocortisone-21 -succinate in rat intestinal perfusate. This figure corresponds to a half-life of 12 h which is significantly longer than the expected transit time to the colon.
  • Gastrointestinal absorption is a function of molecular weight, lipophilicity and polarity; in general, polar, hydrophilic molecules are not well absorbed.
  • the carrier group comprising a substituent represented by Si, for example, are azo linkage linking a carrier, including but not limited to 5 -ASA, and a steroid, thereby generating a mutual prodrug of a drug, such as a steroid.
  • Such mutual prodrugs may provide ideal or favourable physicochemical characteristics for passage through the intestine because of mass, polarity and hydrophilicity.
  • the sterically hindered steroidal esters disclosed in the present application are much less vulnerable towards hydrolysis than hydrocortisone-21 -succinate (see Jhunjhunwala, 1981).
  • the ester may also be attached on one of several different hydroxyl groups on the steroid nucleus.
  • the esters placed at the highly hindered 11 ⁇ axial hydroxyl group (Scheme 2) are more stable than the 21 -esters.
  • the present application discloses novel systems and compounds that are capable of engaging in a two-step process for releasing drugs, such as anti-inflammatory steroids, that target the colon.
  • drugs such as anti-inflammatory steroids
  • the present application also discloses the synthesis of a variety of compounds and their derivatives, including steroidal compounds (e.g. esters of hydrocortisone, dexamethasone or budesonide), nitroimidazoles (e.g. metronidazole), antibiotics, (e.g.
  • quinolines such as nalidixic acid, fluoroquinolones, such as ciprofloxacin or levofloxacin, aminoglycosides, such as amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, streptomycin, tobramycin and apramycin), chemotherapeutics (e.g. leucovorin, topotecan, irinotecan, methotrexate), and antibodies, (e.g. bevacizumab, cetuximab, panitumumab, infliximab).
  • fluoroquinolones such as ciprofloxacin or levofloxacin
  • aminoglycosides such as amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, streptomycin, tobramycin and apramycin
  • chemotherapeutics
  • the present application further discloses the measurement of ring closure kinetics, in vitro screening for intestinal stability using enzymes and intestinal perfusates, drug release in vitro in the presence of colonic bacteria, and therapeutic efficacy, for example, anti-inflammatory efficacy, in appropriate animal models.
  • a series of esters of the anti-inflammatory steroids hydrocortisone, dexamethasone and budesonide at the 21-, 11- and 17 positions are prepared as prodrugs.
  • the application discloses the synthesis of various azo-derivatives of the amino esters.
  • the following Scheme provides representative biologically active agents and drugs, including anti-tumor agents (anthramycin, a sansalvamide derivative, capecitabine), COX-2 inhibitors (resveratrol, curcumin, meloxicam, tenoxicam, piroxicam), antibiotic agents (metronidazole), immunosuppressant agents (cyclosporine) and chemoprotective effect agents (URSO) that may be prepared and employed in the present application:
  • anti-tumor agents anthramycin, a sansalvamide derivative, capecitabine
  • COX-2 inhibitors resveratrol, curcumin, meloxicam, tenoxicam, piroxicam
  • antibiotic agents metalronidazole
  • immunosuppressant agents cyclosporine
  • URSO chemoprotective effect agents
  • analytical methods for the determination or estimation of the rate of ring closure of the amino intermediate compounds are also provided herein. Also provided herein are methods for estimating the rate of ring closure and determining its dependence on mesomeric effects due to substitution in the carrier group, such as the aniline derivatives.
  • a method for measuring the stability of the compounds under aqueous conditions at pH 1-8), in the presence of pancreatic serine proteases (in vitro) and other enzymes present in the gut and, in the presence of rat intestinal fluid.
  • a method for estimating the rate of reduction of these compounds in the presence of microflora in vitro in another aspect, there is provided a method for estimating the in vivo efficacy of the compounds using a mouse model of colonic inflammation.
  • a method for measuring the permeability of the compounds as provided herein using rat perfusion models, which allows the assessment of penetration through the GI tract. For potential colon-targeting, compounds of negligible permeability are employed.
  • Figure 1 is a graph of the concentration of prodrug A versus time at pH 8 (37 0 C).
  • Figure 2 is a graph of the natural log of the prodrug A concentration versus time at pH 8 (37 0 C).
  • Figure 3 is a graph of the concentration of the prodrug A versus time at pH 7.4
  • Figure 4 is a graph of the natural log of the prodrug A concentration versus time in at pH 7.4 (37 0 C).
  • Figure 5 is a profile of the lactamization of aminoprodrug B at pH 9 (37 0 C) showing the disappearance of the prodrug and appearance of cortisone and the quinolone.
  • Figure 6 is a graph of the natural log of the aminoprodrug B concentration versus time at pH 9 (37 0 C).
  • Figure 7 is a profile of the lactamization of amino prodrug B at pH 8 (37 0 C).
  • Figure 8 is a graph of the natural log of the concentration of aminoprodrug B versus time at pH 8.
  • Figure 9 is a profile of the lactamization of aminoprodrug B in PBS at pH 9 (37 0 C) showing the disappearance of the prodrug and appearance of hydrocortisone and quinolone.
  • Figure 10 is a graph of the natural log of the concentration of aminoprodrug B at pH 7.4 (37 0 C).
  • Figure 11 is a profile of the body weight loss percentage of healthy mice (Untreated) and dextran sodium sulphate-induced colitis (DSS-induced colitis) mice treated at 5 mg/Kg dosage.
  • Figure 12 is a profile of the Disease Activity Index (DAI) score from colon segment of DSS-induced colitis mice treated at 5 mg/Kg dosage.
  • DAI Disease Activity Index
  • Figure 13 is a profile of colon length of DSS-induced colitis mice treated at 5mg/Kg dosage.
  • Figure 14 is a profile of thymus weight body weight ratios (IYB W) of healthy mice (Untreated) and DSS-induce colitis mice treated at 5 mg/Kg dosage.
  • Figure 15 shows a rep lotting and reanalysis of the data presented in Figure 11.
  • Figure 11 compared the data sets using a students T test.
  • the data sets have been compared using one-way ANOVA followed by Tukey post test. It also compares the data statistically at days 4-6, as shown in the three blown out graphs. All mice that received DSS lost weight significantly when compared to normal controls. However, when mice were treated with either Prednisolone or Mutual Prodrug 2, this weight loss was decreased as compared to vehicle only controls. There was no statistical difference between the groups that received Prednisolone or Mutual Prodrug 2.
  • Figure 16 shows the replotting and reanalysis of the data presented in figure 12 using the statistical methods described above. All mice that received DSS displayed clinical signs of colitis as compared to the untreated mice. The severity of the disease was significantly reduced in those that received Prednisolone or Mutual Prodrug 2 as compared to those that received vehicle alone. There was no statistical difference between those groups that received Mutual Prodrug 2 or Prednisolone.
  • Figure 17 is a replotting and reanalysis of the data presented in Figure 13, using the statistical methods described above.
  • Colon shortening is a clinical readout for severity of colitis. All mice that received DSS displayed shortening of the colon. The mice that were treated with either Mutual Prodrug 2 or Prednisolone displayed significantly less shortening than those treated with vehicle alone. There was no statistical difference between the Mutual Prodrug 2 or Prednisolone treated groups.
  • Figure 18 shows a replotting and reanalysis of Figure 14 using the statistical methods described above, with additional data showing the thymus weightterminal body weight.
  • the Prednisolone-treated group demonstrated a significant reduction in T/BW ratio as compared to the vehicle-treated mice.
  • the T/BW ratio in the Mutual Prodrug 2 treated group was not statistically different to that of the vehicle-treated group.
  • the synthetic approach is outlined below for dexamethasone (Scheme 4).
  • 2-Nitrocinnamic acid is reduced by catalytic reduction to the aminophenyl propionate and immediately treated with BOC anhydride.
  • the most exposed steroidal hydroxyl group at the 21 position is selectively acylated using
  • N ⁇ '-dicyclohexylcarbodiimide/dimethylaminopyridine (Johnson et al, 1985).
  • the BOC protection may be removed by bubbling anhydrous HCl through a dichloromethane (DCM) solution and the aniline 6 recovered as an HCl salt.
  • the salt is stored at -80 0 C until required.
  • Methyl 3-(2-aminophenyl)propionate spontaneously ring closes even on storage at -20 0 C (Kirby et al., 1979).
  • the isomeric steroidal 21-methoxy-aminophenylacetates 7 and 8 may be obtained via 2-nitration of 3-methoxyphenyl acetic followed by esterif ⁇ cation and reduction ⁇ Scheme 5).
  • the cinnamic acid series may be obtained in a similar manner; nitration of methoxycinammic acid, reduction, BOC protection, esterif ⁇ cation and deprotection.
  • Methoxy groups ortho and para to the amino group may be used to discriminate between steric and electronic influences on lactamization.
  • the more hindered 11-hydroxyl group may be manipulated by protecting the 17 ⁇ - and 21-OH groups (Sloan et al, 1978) before acylation under more forceful conditions (or by using the phenylacetic acid chloride (Spratt et al., 1985)) followed by reduction and deprotection, to yield the 11-phenylacetate.
  • the azo prodrugs of the application may be obtained by esterif ⁇ cation of the steroid with an azo-coupled unit.
  • the 5-ASA-steroid mutual prodrug 9, 5-ASA- dexamethasone, (Scheme 4) may be obtained by esterif ⁇ cation of dexamethasone with compound 10 followed by deprotection of the salicylate.
  • a number of azo- carrier conjugates may be generated to demonstrate intestinal permeability.
  • the kinetics of cyclization may be monitored using UV, HPLC or NMR as appropriate.
  • the quinolone products of spontaneous cyclization of the steroidal propionate esters possess a ⁇ m ax at 340 nm, which allows their measurement in the presence of the ester starting material and steroidal product.
  • the corticosteroids generally have ⁇ max values in the region of 240-250 nm.
  • the reactions may be monitored in buffered D 2 O by 1 H NMR. Although real time analysis is preferable, the reactions may also be monitored by rapid short column HPLC (RT ⁇ 1 min) cooled to inhibit cyclization.
  • the reaction may be monitored in the pH range 5-8 for both the acetate series and the propionate series, at both the 11 and 21 positions.
  • the effect of substitution on the anilino ring may also be determined using this method to determine the optimal therapeutic activities of these compounds and their derivatives.
  • the relative stability of candidate azo-compounds may be evaluated in vitro in the presence of a range of purified pancreatic endopetidases such as the carboxypeptidases (Gilmer et al, 2002). Compounds exhibiting maximum stability may be selected for evaluation in a rat intestinal perfusate model (Friend et al., 1985; Yano et al., 2002). Sections of rat GI tract, obtained from Bioresources, TCD (Ireland) may be gently washed and buffered at appropriate pH. The compounds may be incubated and aliquots withdrawn at intervals. Due to the complex nature of these matrices, remaining drug may be measured by reverse phase HPLC.
  • pancreatic endopetidases such as the carboxypeptidases
  • Half- lives may be determined using Michelis Menten kinetics. Intestinal stability characteristics of selected compounds may be further probed using intestinal homogenates and human CACO-2 cell homogenates. Azoreduction by colonic bacteria in vitro The susceptibility of the azo-bond of candidate compounds to undergo reduction may be assessed using a simple in- vitro bacterial degradation model that employs media inoculated with commercially available Lactobacillus culture (Azad Khan et al, 1983) (this organism expresses similar azo-reductases to colonic bacteria). The medium may be supplemented with the coenzymes NADPH and FAD. The reduction may be further investigated using rat or guinea pig colonic perfusate that has been saturated with CO 2 (see for example Yano, 2002). Anti-inflammatory activity
  • the efficacy of the prodrugs in vivo may be assessed in an animal model.
  • DSS dextran sodium sulphate-induced
  • the balb/c mouse is a suitable strain to use for this experiment, as it is known to be susceptible to oral DSS, resulting in colitis (Egger et al., 2000).
  • Female mice 6-8 weeks of age may be used to ensure the animals are mature and comparable in weight.
  • the intestines may be removed for histological examination and RNA extraction, therefore a further 3 untreated mice are required to practice dissection, histology and RNA extraction techniques.
  • a section of the colon may be fixed in 10% zinc-formalin and embedded in paraffin.
  • a transverse section may be stained routinely with haematoxylin and eosin.
  • the severity of mucosal injury is graded on a scale of 0-III: grade 0: normal; grade I: distortion and/or destruction of the bottom third of the glands; grade II: erosions/destruction of bottom two-thirds or loss of all glands with remaining surface epithelium; grade III, loss of all glands (Egger et al., 2000; Festing et al., 2002; Egger et al., 1997).
  • Reverse transcriptase polymerase chain reaction may be carried out on extracted RNA to determine levels of NF ⁇ B-dependent pro -inflammatory cytokines IL-I and TNF- ⁇ , which are up-regulated in DSS-induced colitis (Egger et al., 2000).
  • the experiments may be repeated with different concentrations of pro-drug and different numbers of doses if necessary.
  • the prodrugs have been designed to be excluded from the absorption process, thus permitting their transit to the colon.
  • Compounds for further evaluation may be selected on the basis of their poor permeability characteristics.
  • the GI behaviour of the compounds may be evaluated first using the everted intestinal sac (Wilson and Wiseman, 1954), which permits direct evaluation and comparison of drug transfer across the in vitro barrier and the intestinal perfusion model described below.
  • the mesenteric vein is cannulated with an appropriate size of Silastic or polyethylene tubing and venous blood is collected in heparinized (or equivalent), calibrated centrifuge tubes. After gentle washing of the intestinal lumen with isotonic phosphate buffer at 37 0 C, drug solution is perfused into the proximal section of the segment allowed to flow through the segment at 0.2 ml/min and collected at the distal end of the lumenal segment.
  • the isolated intestine is kept warm and moist by frequent application of warm (37 0 C) saline to a gauze pad covering the intestine, which is covered by a dental rubber dam. A small lamp is also used to maintain the preparation at 37 0 C.
  • the drug is perfused through the intestine for up to 120 minutes with perfusate collection at regular intervals. Perfusate is assayed and drug disappearance from perfusion fluid is monitored over the time course of the experiment.
  • the experiments provide an evaluation of the methods as a means of targeting the colon with anti-inflammatory steroids including novel mutual prodrugs of the two principal therapies used in IBD.
  • a method for the synthesis of several new families of novel steroidal esters bearing anilines and azo carriers provides comprehensive study of the cyclization kinetics of aminophenyl acetate and propionate esters under simulated physiological conditions and insight into the electronic and steric ring substitution effects on closure kinetics.
  • the method provides a determination of the aqueous, enzymatic and intestinal stability of 21 and 11- corticosteroid esters.
  • the method allows the determination of the rate of reduction of steroid aza conjugates and dependence on steroid and ring substitution effects.
  • A is hydrogen or is a residue of a hydroxyl-bearing drug
  • Z is selected from the group consisting of -C(O)-, -S(O)-, -OC(O)-, -OC(O)NR 13 -, -S-C(O)-, -SC(O)NR 13 -, - C(O)NR 13 -, -NR 13 C(O)NR 13 -, -OS(O)-, -OS(O) 2 -, -S(O) 2 NH- and -OPO(OH)-; each R 1 , R 2 , R 5 and R 6 is independently hydrogen or are each independently selected from the group consisting of (Ci- 3 )alkyl, (C 3 _io)cycloalkyl, (C 3 _io)cycloalkyl(Ci- 3 )alkyl, aryl, aryl(Ci- 3 )alkyl, heteroaryl, heteroaryl(Cii
  • R 1 ⁇ and R 12 are taken together to form an optionally substituted heterocyclic ring.
  • R 11 and R 12 are taken together to form acetonidyl-4-one.
  • R 11 is C(O)O-R 10
  • each of R 12 and R 10 is hydrogen.
  • A is hydrogen or is a residue of a hydroxyl-bearing drug
  • each R 1 , R 2 , R 5 and R 6 is independently selected from the group consisting of hydrogen, (Ci- 3 )alkyl, (C 3 -io)cycloalkyl, (C 3 -io)cycloalkyl(Ci- 3 )alkyl, aryl, aryl(Ci- 3 )alkyl, heteroaryl, heteroaryl(Ci-3)alkyl, cyano, halo, hydroxy, (Ci-3)alkoxy, aryloxy and heteroaryloxy, each substituted or unsubstituted; each R 3 and R 4 are independently hydrogen or are each independently selected from the group consisting of (Ci- 3 )alkyl, (C 3 -io)cycloalkyl, (C 3 -io)cycloalkyl(Ci- 3 )alkyl, aryl, aryl(Ci- 3 )
  • R 7 and R 8 are hydrogen, and c is 1 or 2. In another variation, a and b are both 0, and c is 1 or 2. In yet another variation, R 9 is hydrogen; in another variation, R 7 and R 8 are hydrogen, R 9 is hydrogen, and a and b are both 0.
  • A is hydrogen or is a residue of a hydroxyl-bearing drug
  • A is hydrogen or is a residue of a hydroxyl-bearing drug
  • R 5 and R 6 are independently hydrogen or are independently selected from the group consisting of (Ci- 3 )alkyl, (C 3 _io)cycloalkyl, (C 3 _io)cycloalkyl(Ci- 3 )alkyl, aryl, aryl(Ci- 3 )alkyl, heteroaryl, heteroaryl(Ci- 3 )alkyl, cyano, halo, hydroxy, (Ci- 3 )alkoxy, aryloxy and heteroaryloxy, each substituted or unsubstituted;
  • R 7 is hydrogen or is (Ci-3)alkyl;
  • R 8 is hydrogen or is (C 1 - 3 )alkyl; or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers.
  • R 5 and R 6 are hydrogen; in another embodiment, R 5 , R 6
  • a or the hydroxyl-bearing drug of any of the disclosed compounds is selected from the group consisting of an anti-inflammatory drug, an anti-cancer drug, an imaging agent; a vaccine, an antigen, an anti-infective drug, a peptide, an antisense molecule and a protein.
  • the hydroxyl-bearing drug is an antiinflammatory drug.
  • the anti-inflammatory drug is a steroid.
  • the steroid is selected from the group consisting of hydrocortisone, dexamethasone, budesonide esterified at the 21-, 11- and 17- positions, respectively and prednisolone.
  • R 11 and the linker comprising the azo group is 1 ,4-di-substituted (i.e., para) on the phenyl ring.
  • R 12 is H.
  • a is 0; or a and b are both 0.
  • R 9 is H.
  • a or the hydroxyl-bearing drug of any of the disclosed compounds is selected from the group consisting of nitroimidazoles, quinolines such as nalidixic acid, fluoroquinolones, such as ciprofloxacin or levofloxacin, aminoglycosides, such as amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, streptomycin, tobramycin and apramycin, leucovorin, topotecan, irinotecan, methotrexate, bevacizumab, cetuximab, panitumumab and infliximab.
  • the drug is an amine bearing drug (i.e., a drug with an amine group) such as celecoxib, or a thiol bearing drug.
  • the compound of present application is selected from the group consisting of:
  • a pharmaceutical composition comprising a therapeutically effective amount of any of the disclosed compounds provided herein and a pharmaceutically acceptable excipient.
  • a method for decreasing NF ⁇ ;B DNA-binding activity in a patient comprising administering a therapeutically effective amount of any compound or composition of the present application to the patient.
  • the therapeutically effective amount is effective to reduce, alleviate or treat inflammatory bowel disease (IBD).
  • the therapeutically effective amount is effective to reduce, alleviate or treat ulcerative colitis or Crohn's disease.
  • a method of treating inflammatory bowel disease comprising administering a therapeutically effective amount of any compound or composition of the present application to a mammal in need of such treatment.
  • a method of treating Crohn's disease or ulcerative colitis comprising administering a therapeutically effective amount of any compound or composition of the present application to a mammal in need of such treatment.
  • the amount of compound or composition administered is enough to maintain remission.
  • 5ASA-5ASA is administered.
  • a method of reducing, alleviating or treating acute ulcerative colitis comprising administering a therapeutically effective amount of 5ASA-steroid, wherein steroid is hydrocortisone, dexamethasone, budesonide esterified at the 21-, 11- and 17- positions, respectively, or prednisolone.
  • steroid is prednisolone.
  • a method of treating Collagenous colitis, Lymphocytic colitis, Ischaemic colitis, Diversion colitis, Behcet's syndrome, Infective colitis, or Indeterminate colitis comprising administering a therapeutically effective amount of any compound or composition of the present application to a mammal in need of such treatment.
  • a method of treating Amebiasis, Clostridium Difficile Infection, Pseudomembranous colitis, Diverticulitis, Gastroenteritis, Gastrointestinal Cancers, or Irritable Bowel Syndrome (IBS) comprising administering a therapeutically effective amount of any compound or composition of the present application to a mammal in need of such treatment.
  • a method for treating or alleviating an inflammatory condition in a mammal comprising delivering an effective amount of a COX2 inhibitor to the colon, wherein the COX2 inhibitor is the residue of a hydroxyl bearing drug of any of the compound or compositions as provided herein.
  • a method for treating gastrointestinal cancer in a mammal comprising delivering an effective amount of a COX2 inhibitor to the colon, wherein the COX2 inhibitor is the residue of a hydroxyl bearing drug of any of the compound or compositions as provided herein.
  • compositions comprising pharmaceutically acceptable excipients and a therapeutically effective amount of at least one compound of this application.
  • compositions of the compounds of this application, or derivatives thereof may be formulated as solutions or lyophilized powders for parenteral administration.
  • Powders may be reconstituted by addition of a suitable diluent or other pharmaceutically acceptable carrier prior to use.
  • the liquid formulation is generally a buffered, isotonic, aqueous solution.
  • suitable diluents are normal isotonic saline solution, 5% dextrose in water or buffered sodium or ammonium acetate solution.
  • Such formulations are especially suitable for parenteral administration but may also be used for oral administration.
  • Excipients such as polyvinylpyrrolidinone, gelatin, hydroxycellulose, acacia, polyethylene glycol, mannitol, sodium chloride, or sodium citrate, may also be added. Alternatively, these compounds may be encapsulated, tableted, or prepared in an emulsion or syrup for oral administration.
  • Pharmaceutically acceptable solid or liquid carriers may be added to enhance or stabilize the composition, or to facilitate preparation of the composition. Liquid carriers include syrup, peanut oil, olive oil, glycerin, saline, alcohols or water.
  • Solid carriers include starch, lactose, calcium sulfate, dihydrate, terra alba, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin.
  • the carrier may also include a sustained release material such as glyceryl monostearate or glyceryl distearate, alone or with a wax.
  • the amount of solid carrier varies but, preferably, may be between about 20 mg to about 1 g per dosage unit.
  • the pharmaceutical preparations are made following the conventional techniques of pharmacy involving milling, mixing, granulation, and compressing, when necessary, for tablet forms; or milling, mixing, and filling for hard gelatin capsule forms.
  • the preparation may be in the form of a syrup, elixir, emulsion, or an aqueous or non-aqueous suspension.
  • a liquid formulation may be administered directly p.o. or filled into a soft gelatin capsule.
  • Suitable formulations for each of these methods of administration may be found in, for example, Remington: The Science and Practice of Pharmacy, A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, Pa.
  • the above compound or a pharmaceutically acceptable salt thereof, optionally in the form of a single stereoisomer or mixture of stereoisomers thereof.
  • alkyl is a straight, branched, saturated or unsaturated, aliphatic group having a chain of carbon atoms, optionally with oxygen, nitrogen or sulfur atoms inserted between the carbon atoms in the chain or as indicated.
  • a (Ci_Cio)alkyl includes alkyl groups that have a chain of between 1 and 10 carbon atoms, and include, for example, the groups methyl, ethyl, propyl, isopropyl, vinyl, allyl, 1-propenyl, isopropenyl, ethynyl, 1-propynyl, 2-propynyl, 1,3-butadienyl, penta-l,3-dienyl, and the like.
  • An alkyl group may also be represented, for example, as a -(CR 1 R 2 ) a - group where R 1 and R 2 are independently hydrogen or are independently substituted as provided herein; and for example, a is 0, 1 or 2.
  • An alkyl as noted with another group such as an aryl group, represented as
  • arylalkyl for example, is intended to be a straight, branched, saturated or unsaturated aliphatic divalent group with the number of atoms indicated in the alkyl group (as in (Ci_Cio)alkyl, for example) and/or aryl group or when no atoms are indicated means a bond between the aryl and the alkyl group.
  • Nonexclusive examples of such group include benzyl, phenylethyl and the like.
  • alkylene is a straight, branched, saturated or unsaturated aliphatic divalent group with the number of atoms indicated in the alkyl group; for example, a -(Ci_C 3 )alkylene- or -(Ci_C 3 )alkylenyl-.
  • aryl is a monocyclic or bicyclic aromatic hydrocarbon group having 5 to 8 atoms in the ring, such as a phenyl.
  • the monocyclic aryl groups are typically are 5 to 7 membered rings, and the bicyclic aryl groups are typically 7 to 8 membered rings.
  • heteroaryl means an aryl group containing from, for example, about 3 to about 30 atoms, preferably from about 6 to about 18 atoms, more preferably from about 6 to about 14 atoms, and most preferably from about 6 to about 10 atoms and from 1 to 3 heteroatoms (e.g., N, O or S).
  • Examples of such groups include pyrrolyl, imidazolyl, pyrazolyl, furanyl, oxazolyl, isooxazolyl, thiofuranyl, thiazolyl, isothiazolyl, indolyl, isoindolyl, benzofuranyl, quinolinyl, pyridinyl, pyridazinyl, pyrazinyl, triazolyl and benzotriazolyl.
  • a “cyclyl” such as a monocyclyl or polycyclyl group includes monocyclic, or linearly fused, angularly fused or bridged polycycloalkyl, or combinations thereof. Such cyclyl group is intended to include the heterocyclyl analogs.
  • a cyclyl group may be saturated, partially saturated or aromatic.
  • Halogen or “halo” means fluorine, chlorine, bromine or iodine.
  • heterocyclyl or “heterocycle” is a cycloalkyl group wherein one or more of the atoms forming the ring is a heteroatom that is a N, O or S.
  • the cycloalkyl may be saturated, partially saturated or aromatic.
  • heterocyclyl examples include piperidyl, 4-morpholyl, 4-piperazinyl, pyrrolidinyl, 1,4-diazaperhydroepinyl, acetonidyl-4- one, 1,3-dioxanyl, thiophenyl, furanyl, pyrrolyl, pyrazolyl, pyridinyl, pyrimidinyl pyridazinyl, pyranyl and the like.
  • alkoxy includes linear or branched alkyl groups that are attached to divalent oxygen.
  • the alkyl group is as defined above. Examples of such substituents include methoxy, ethoxy, t-butoxy, and the like.
  • alkoxyalkyl refers to an alkyl group that is substituted with one or more alkoxy groups.
  • heteroaryloxy refers to a heteroaryl group that is substituted with one or more alkoxy groups.
  • aryloxy refers to an aryl group that is attached to an oxygen, such as phenyl-O-, etc.
  • a divalent group is represented by a group -Z- as described herein, or generically as -A-B-, as shown below for example, it is intended to also represent a group that may be attached in both possible permutations, as noted in the two structures below.
  • a divalent group such as the group "-NR 13 C(O)-"
  • the group is intended to also include both the divalent group -NR 13 C(O)- and also the divalent group -C(O)NR 13 -.
  • a "hydroxyl-bearing drug” means a drug or a biologically active compound that is functionalized or that is substituted with a hydroxyl (i.e., -OH) group. Accordingly, the residue of a hydroxyl-bearing drug is the component of the drug or the biologically active compound without the hydroxyl group.
  • Drugs employed in the present application may include an anti-inflammatory drug, an anti-cancer drug, an imaging agent, particularly as used in the imaging of colon diseases; a vaccine, an antigen, an anti-infective drug, a peptide, an antisense molecule, and a protein.
  • Such drugs can include, but are not limited to steroidal compounds (e.g.
  • esters of hydrocortisone, dexamethasone or budesonide nitroimidazoles (e.g. metronidazole), antibiotics, (e.g.quinolines such as nalidixic acid, fluoroquinolones, such as ciprofloxacin and levofloxacin), chemotherapeutics (e.g. leucovorin, topotecan), and antibodies, (e.g. bevacizumab, cetuximab and panitumumab).
  • “Pharmaceutically acceptable salts” means salt compositions that is generally considered to have the desired pharmacological activity, is considered to be safe, non-toxic and is acceptable for veterinary and human pharmaceutical applications.
  • Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, and the like; or with organic acids such as acetic acid, propionic acid, hexanoic acid, malonic acid, succinic acid, malic acid, citric acid, gluconic acid, salicylic acid and the like.
  • Pro-drug or “prodrug” as used herein, means a bioprecursor or pharmaceutically acceptable compound that may be convertible or degradable in the body, specifically in the colon, to produce a biologically active compound(s) of the invention (for example, the intermediate aniline or the lactam and the active drug).
  • a biologically active compound(s) of the invention for example, the intermediate aniline or the lactam and the active drug.
  • the compounds of the present application may be reduced by an in vivo azoreductase such as microflora azoreductase.
  • Therapeutically effective amount means a drug amount that elicits any of the biological effects listed in the specification.
  • a therapeutically effective amount also means an amount of the composition comprising a compound/agent or pro-drug, as disclosed herein, that is useful or capable of supporting an observable change in the level of one or more biological activity characteristic of a compound/agent or pro-drug, or a dosage amount that is sufficient to impart a beneficial effect.
  • Such beneficial effect may comprise an amelioration of a symptom for the recipient or patient.
  • the therapeutically effective amount may be related to the healing or curing of disease. As is known in the art, a number of considerations may be taken into account to determine the therapeutically effective amount.
  • Substituted or unsubstituted or “optionally substituted” means that a group such as, for example, alkyl, aryl, heterocyclyl, (Ci-C 8 )cycloalkyl, heterocyclyl(Ci-Cs)alkyl, aryl(Ci-Cs)alkyl, heteroaryl, heteroaryl(Ci-C 8 )alkyl, and the like, unless specifically noted otherwise, may be unsubstituted or, may substituted by 1, 2 or 3 substituents selected from the group such as halo, nitro, trifluoromethyl (-CF3), trifluoromethoxy, methoxy, carboxy, -NH 2 , -OH, -SH, -SCH 3 , -NHCH 3 , -N(CH 3 ) 2 , -SMe, cyano and the like.
  • protective groups may be introduced and finally removed.
  • Suitable protective groups for amino, hydroxy, and carboxy groups are described in Greene et al, Protective Groups in Organic Synthesis, Second Edition, John Wiley and Sons, New York, 1991. Standard organic chemical reactions can be achieved by using a number of different reagents, for examples, as described in Larock: Comprehensive Organic Transformations, VCH Publishers, New York, 1989.
  • the compounds of this application can be synthesized by the steps outlined in Schemes 8-11 below.
  • the schemes demonstrate the preparation of different homo logs of the pro-drugs.
  • LRMS was performed on a micromass mass spectrophotometer (EI mode) at the Department of Chemistry, Trinity College. Flash chromatography was performed on Merck Kieselgel 60 particle size 0.040-0.063 mm thin layer chromatography (TLC) for which Rf values are quoted, was performed on silica gel Merck F-254 plates. Compounds were visually detected by absorbance at 254 nm +/or vanillin staining.
  • DCC Dicyclohexylcarbodiimide
  • DMAP 4-(dimethylamino)pyridine
  • DCU dicyclohexylurea
  • 5-Nitrosalicylic acid dioxin-4-one 5-Nitrosalicylic acid (20 g, 109.2 mmol) was placed in a 500 ml round bottom flask equipped with a magnetic stirrer and a reflux condenser, 200 ml of trifluoroacetic acid was added followed by trifluoroacetic anhydride (45.54 ml, 327.7 mmol) and dry acetone (16.04 ml, 218.4 mmol). The reaction mixture was left at reflux for two hours. Further 8.02 ml of dry acetone was dropped to the boiling solution every hour (1 eq per hour) until reaction is complete within eight hours.
  • the 2-nitrophenyl acid methyl ester (1 g, 5.13 mmol) was dissolved in 20 mL of EA and 0.3 g of Pd/C 10% was suspended into the solution, the reaction mixture was under hydrogen atmosphere at room temperature. After 2h the reaction was completed by TLC analysis (DCM: EA 60; 40) the Pd/C 10% was removed by filtration and the solvent by reduced pressure; to yield a colorless oil 0.67 g (67%).
  • 2-(4-Hydroxy-phenylazo)-phenyl-acetic acid methyl ester The 2-aminophenyl acid methyl ester (3 g, 0.018 mol) was dissolved in water
  • the azo carrier (0.5 g, 0.0019 mol,) was added to a solution of hydrocortisone (0.34 g, 0.0009 mol,) and triphenylphosphine (0.74 g, 0.0028 mol) in dry THF, under nitrogen atmosphere the temperature was raised to 40 0 C and diisopropylazodicarboxylate 95% (DIAD) (0.57 g, 0.0028 mol) was added dropwise over 12 minutes with vigorous stirring. The mixture was stirred at 40 0 C over Ih and reaction was left overnight at room temperature under nitrogen atmosphere. The volatiles were removed under reduced pressure to afford the product as orange oil.
  • DIAD diisopropylazodicarboxylate 95%
  • the first order rate constant for disappearance can be obtained from a plot of the natural log of remaining ester versus time.
  • the calculated half-life of prodrug A is 2.2 hours; the calculated half-life of prodrug B is 1.4 hours.
  • Prednisolone and DSS were purchased from Sigma- Aldrich laboratories.
  • BALB/c strain mice were from the Bioresources Unit (Trinity College Dublin).
  • mice were housed in individually ventilated and filtered cages under positive pressure (Tecniplast, Nothants, UK). Food and water were supplied ad libitum. All animal experiments were performed in compliance with Irish Department of Health and Children regulations and approved by the Trinity College Bioresources ethical review board. Statistical analysis
  • mice All in vivo experiments were performed with six mice per group. Difference between groups was analyzed by Student's t-test. Colitis scores were analyzed by Mann- Whitney test. P values ⁇ 0.05 were considered significant. Results on the DIA, C/B and T/B MPO were expressed as means ⁇ S. E. Preparation of inflammatory bowel disease model mice
  • DSS 35- 50,000 kDa; MP Biomedicals, OH
  • Fresh DSS solution was provided every second day.
  • BALB/c mice were exposed to 5% DSS for six days. The mice were checked every day for morbidity and weight recorded. Induction of colitis was determined by weight loss, fecal blood and, upon autopsy, length of colon. Blood in feces was detected using a Hemdetect occult blood detection kit (Dipro, Austria).
  • DAI disease activity index
  • Tissue Damage Score 0, No mucosal damage; 1, Partial (up to 50%) loss of crypts in large areas; 2, Partial to total 50-100% loss of crypts in large areas, epithelium intact; 3, Total loss of crypts in large areas and epithelium lost.
  • the DSS induced colitis mice weighting about 18-20 grams were allowed 5% DSS water ad libitum and 100 ⁇ L of prednisolone and prodrug 2 in 1% solution cremophor and ethanol in water suspension (equivalent to 5 mg of prednisolone per Kg or mouse body weight) was orally administrated, twice a day (every twelve hours) to the colitis mice.
  • the mice were sacrificed by cervical dislocation. The distal colon segment and the thymus were removed and washed to remove contents of the colon.
  • Anti-inflammatory effects were studied by comparison of the distal colon length between healthy mice, untreated mice, prednisolone treated mice and prodrug treated mice. IBD causes shortering of the colon. Thus anti-inflammatory effects the less damage was observed on the colon tissue and less reduction was cause on the distal colon length.
  • thymus weight body weight ratios T:BW
  • Untreated mice DSS-induced colitis mice treated at 5 mg/Kg dosage
  • the PRED group had significant reduction in thymus :B W ratio versus the Untreated mice (P ⁇ 0.05).
  • the mean thymus weight in the prodrug treatment groups was not statistically different to the untreated mice.
  • a novel colon-specific steroid prodrug enhances sodium chloride absorption in rat colitis.

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Abstract

L'invention porte sur des composés, des compositions et des procédés pour diminuer l'activité de liaison à NFkB ADN dans un patient, comprenant l'administration d'une quantité thérapeutiquement efficace d'un composé ou d'une composition de l'application au patient pour réduire, soulager ou traiter diverses maladies gastro-intestinales, telles que la maladie intestinale inflammatoire (IBD).
PCT/EP2008/058338 2007-06-29 2008-06-27 Ciblage de promédicaments diazo pour le traitement de maladies gastro-intestinales WO2009003970A1 (fr)

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AU2008270363A AU2008270363A1 (en) 2007-06-29 2008-06-27 Targeting diazo prodrugs for the treatment of gastrointestinal diseases
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CN200880022191A CN101801919A (zh) 2007-06-29 2008-06-27 治疗胃肠疾病的靶向重氮前药
JP2010513951A JP2010531845A (ja) 2007-06-29 2008-06-27 胃腸疾患の治療のための標的指向化ジアゾプロドラッグ
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Publication number Priority date Publication date Assignee Title
WO2010072734A3 (fr) * 2008-12-23 2010-08-19 The Provost Fellows And Scholars Of The College Of The Holy And Undivided Trinity Of Queen Elizabeth Near Dublin Promédicaments ciblants et compositions destinés au traitement de maladies gastro-intestinales
WO2020172632A1 (fr) * 2019-02-21 2020-08-27 The Regents Of The University Of Colorado, A Body Corporate Composés azoïques antimicrobiens et utilisations associées
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CN106032358A (zh) * 2015-03-18 2016-10-19 浙江九洲药业股份有限公司 一种柳氮磺吡啶的合成工艺
EP3484483B1 (fr) * 2016-07-13 2023-03-08 Kaleido Biosciences, Inc. Compositions de glycane et leurs procédés d'utilisation
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MA52266A (fr) * 2018-03-07 2021-01-13 Amgen Europe Gmbh Procédés et compositions de promédicaments isoindoline-1,3-dione et isoindole utiles pour le traitement du cancer, de la rectocolite hémorragique et de maladies inflammatoires associées
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1930379A2 (fr) * 2006-12-05 2008-06-11 Everlight USA, Inc. Composition réactive de colorant rouge et son application

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1930379A2 (fr) * 2006-12-05 2008-06-11 Everlight USA, Inc. Composition réactive de colorant rouge et son application

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
MICHAEL G HUTCHINGS ET AL: "Syntheses of 2-aryl-3-cinnolinones by cyclisation of diarylazo compounds", TETRAHEDRON LETTERS, ELSEVIER, AMSTERDAM, vol. 30, no. 28, 1 January 1989 (1989-01-01), pages 3715 - 3718, XP009107880, ISSN: 0040-4039 *
SINHA, KUMRIA: "Colonic drug delivery: prodrug approach", PHARMACEUTICAL RESEARCH, vol. 18, no. 5, 2001, pages 557 - 564, XP002501855 *
VON STUDNITZ ET AL: "Comparative studies of 3-hydroxy-4-methoxymandelic acid and 3-methoxy-4-hydroxymandelic acid", CLINICA CHIMICA ACTA, ELSEVIER BV, AMSTERDAM, NL, vol. 12, no. 3, 1 September 1965 (1965-09-01), pages 330 - 334, XP023391283, ISSN: 0009-8981, [retrieved on 19650901] *

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US11883422B2 (en) 2015-04-23 2024-01-30 Dsm Nutritional Products, Llc Glycan therapeutic compositions and related methods thereof
US11540909B2 (en) 2017-08-21 2023-01-03 The Regents Of The University Of Colorado, A Body Corporate Azobenzene polymer network, and uses thereof for biofilm removal and control over cell attachment
WO2020172632A1 (fr) * 2019-02-21 2020-08-27 The Regents Of The University Of Colorado, A Body Corporate Composés azoïques antimicrobiens et utilisations associées
CN113004355A (zh) * 2019-12-20 2021-06-22 复旦大学 呋喃酮糖苷类化合物、其药物组合物、制备方法及应用
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JP2010531845A (ja) 2010-09-30

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