MXPA01001243A - Compounds and compositions for delivering active agents - Google Patents

Abstract

Compounds and compositions for the delivery of active agents are provided. Methods of administration and preparation are provided as well.

Description

COMPOUNDS AND COMPOSITIONS TO RELEASE ACTIVE AGENTS FIELD OF THE INVENTION The present invention relates to compounds for releasing active agents, such as biologically or chemically active agents, to a target. These compounds are well suited to form non-covalent mixtures with active agents for oral, intracolonic, or other routes to animals. Methods for the preparation and administration of such compositions are also described.

BACKGROUND OF THE INVENTION Conventional means for releasing active agents are often severely limited by biological, chemical, and physical barriers. Typically, those barriers are imposed by the environment through which liberation occurs, the environment of the goal for liberation, and / or the objective itself. Biologically and chemically active agents are particularly vulnerable to such barriers. In the administration to animals of biologically active and chemically active pharmacological and therapeutic agents, barriers are imposed by the body. Examples of physical barriers are the skin, Ref: 126792 Lipid bilayers and several membranes of organs that are relatively impervious to certain active agents, but must be traversed to reach the target, such as a circulatory system. Chemical barriers include, but are not limited to, pH variations in the gastrointestinal tract (Gl) and degrading enzymes. These barriers are of particular significance in the design of oral administration systems. The oral testing of many biological and chemically active agents would be the route of choice for administration to animals if there were no biological, chemical and physical barriers. Among the numerous agents that are not typically suitable for oral administration are biologically or chemically active peptides, such as calcitonin and insulin; polysaccharides, and in particular mucopolysaccharides including, but not limited to, heparin; heparinoids; antibiotics; and other organic substances. These agents can quickly become ineffective or be destroyed in the gastrointestinal tract by acid hydrolysis, enzymes and the like. In addition, the size and structure of macromolecular drugs can prohibit absorption. The first methods of administering orally vulnerable pharmacological agents depended on the co-administration of adjuvants (for example, resorcinols and non-ionic surfactants such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether) to artificially increase the permeability of intestinal walls, as well as co-administration of enzymatic inhibitors (eg, pancreatic trypsin inhibitors, diisopropyl fluorophosphate (DFF) and trasilol) to inhibit enzymatic degradation. Liposomes have also been described as a drug delivery system for insulin and heparin. See, for example, US 4,239,754; Patel et al., (1976), FEBS Letters, Vol 62, pg. 60; and Hashimoto et al. (1979), Endocrinology Japan, Vol. 26, pg. 337. However, the use of broad spectrum of such drug delivery systems is impeded because: (1) the systems require toxic amounts of adjuvants or inhibitors; (2) charges, ie, suitable low molecular weight active agents are not available; (3) the systems exhibit poor instability and inadequate shelf life; (4) systems are difficult to manufacture; (5) the systems do not protect the active agent (charge); (6) the systems adversely alter the active agent; or (7) the systems do not allow or promote the absorption of the active agent. More recently, artificial polymer microspheres of mixed amino acids (proteinoids) have been used to release drugs. For example, U.S. Patent 4,925,673 discloses compounds in the form of drug-containing proteinoid microspheres, as well as methods for their preparation and use. These proteinoid microspheres are useful for the release of a number of active agents. In addition, certain modified amino acids have been used to release drugs. See, for example, US 5,629,020; US 5,643,957, US 5,650,386; and US 5,776,888. However, there is still a need for simple, inexpensive delivery systems that can be readily prepared and that can release a wide range of active agents by several routes.

BRIEF DESCRIPTION OF THE INVENTION Compounds and compositions that are useful in the delivery of active agents are provided. The compounds comprise the following compounds or salts thereof.

Compound # N m X 1 3 CH20 2-OH 2 3 CH20 4 -OH 3 3 0 2 -NH2, 5- F 4 2 0 2 -NH2, 5-F 5 3 0 2 -NH2, 5-C1 Compound # N m X 6 2 0 2-NH 2, 3, 5-Cl 2 0 2-NHMe 3 I 4-OH 9 3 1 3-OH 10 3 or 2-NHMe 11 2 or 2-OH, 3-F; 5-Cl 12 2 or 2-OH, 3-C1; 5-F 13 2 or 2-OH, 3,5-Me 14 3 or 2-OH, 3,5-Me 2 or 2-OH, 3-Br, 5-Cl 16 3 2 2-OH 17 2 2 2-OH 18 2 0 2-OH, 3, 5-F 19 3 or 2-OH, 3, 5-F 2 or 2-OH, 5-F 21 3 or 2-OH, 5-F 2 2 or 2-NHAc 3 3 or 2-S03Na 4 3 or 2-OH, 3-Me, 5-F 5 3 or 2 -OH, 3-Me, 5-Cl 04 3 or 2-OH, 4-Ce 33 2 or 2-OH, 3-Me, 5-Cl Compound # n X 26 7 2-OH, 5-Me 27 6 2-OH 28 8 2-OH, 3, 5-Cl 29 7 2-OH, 3, 5-Cl 8 2-OH, 4-Me 31 7 2-CH2OH 32 4 2-OH, 4-Me 33 7 2 -OH, 4 -Me 34 7 2-OH, 5-F 8 2-OH, 5-F 119 3 2 -OH, 5-Cl 120 5 2 -OH, 5-Cl 121 6 2-OH, 5-Cl 122 7 2-OH, 5-Cl 123 8 2 -OH, 5-Cl 124 1 2-OH, 5-Cl 125 2 2 -OH, 5-Cl 126 4 2 -OH, 5-Cl 127 9 2-OH, 5-Cl Compound # n X 128 10 2-OH, 5-Cl 129 11 2-OH, 5-Cl 130 12 2-OH, 5-Cl 131 7 2-OH, 3, 4- F 132 7 2-OH, 4-F Compound # N m X 36 7 0 2-OH, 3-NH 2, 5-N02 37 5 0 2-OH, 4-Cl 38 7 CH20 4-OH 39 7 0 2-NH 2, 5-F 40 7 0 2-NH 2, 5-Cl 41 7 0 2-OH, 3, 5-F 42 7 0 2-OH, 3, 4-F 43 7 0 2-NHMe 44 7 0 2-OH, 4-F 45 7 0 2-OH, 3-F, 5-Cl 46 7 1 4-OH 47 7 0 2-OH, 3-C1, 5-F 8 7 0 2-OH, 3-Br, 5-Cl Compound # N m X 49 7 0 2-OH, 3,5- Me 50 7 0 2-OMe, 6-C1 51 7 0 2-OH, 6-C1 52 7 1 3-OH 53 7 2 2-OH 54 7 0 2 -OH, 5-F 55 7 0 2-OH, 3-Me, 5-Cl 56 7 0 2-OH, 3-Me, 5-F 57 9 0 2-OH, 5-Cl 85 9 0 2-F 86 5 or 2-F 87 ll or H 10 or 2-F 89 5 or H 90 3 or 2-OCH 3 91 3 or 2-CH 3 92 3 or 2-F 93 3 or H 94 9 or 2-OCH 3 95 11 or 2-CH 3 6 11 or 2-OCH 3 7 11 or 2-F Compound # nm X 99 9 0 2-CH 3 100 9 0 H 101 5 0 2-CH3 102 1 0 2-OH, 4-OMe 103 2 0 2-OH 105 3 0 2-OH, 5-Cl 106 3 0 2-OH, 4-OMe 107 5 0 2-OH, 4-OMe 108 9 0 2-OH, 4-C s 109 11 0 2-OH, 4-C s 110 1 0 H 111 1 0 2-CH3 112 1 0 2-avfe 113 1 0 2-F 114 1 0 2-OH, 5-Cl 116 4 0 2-OH, 5-Cl 117 5 0 2-OH, 5-Cl 118 6 0 2-OH, 5-Cl Compound 58: Compound 59: Compound 60: X = F Compound 61: X = 0H Compound 62: X = SO Compound 63: X = S02 ^^^^^^^^^ Compound 64: Compound 65: X = C1; Y =? Me Compound 66: X = F; Y = OH Compound 67: X = F; Y = OMe Compound 68: X = 0Me; Y = OH Compound 69: n = l Compound 70: n = 2 Compound 71: Compound 72 Compound 73 Compound 74: X = Me Compound 75: Compound 76: X = Y = OMe; z = H; n = l Compound 77: X = OH; Y = Z = C1; n = l Compound 78: n = 2 Compound 134: n = 3 Compound 79: Compound 80: Compound 81: Compound 82 Compound 83: Compound 84: R = C (O) -NH- (CH2) 7COOH Compound 135: The compositions of the present invention comprise at least one active agent, preferably a biologically or chemically active agent, and at least one of the compounds, or salts thereof, of the present invention. Methods for the preparation and administration of such compositions are also provided. Compositions comprising the compounds and active agents have utility in the release of active agents to selected biological systems and in the increase or improvement of the bioavailability of the active agent as compared to the administration of the active agent alone.

DETAILED DESCRIPTION OF THE INVENTION The compositions of the present invention include an active agent and a release agent. These compositions can be used to release various active agents through various biological, chemical and physical barriers and are particularly suitable for releasing active agents that are subject to environmental degradation. Other advantages of the present invention include the use of cheap, easy-to-prepare raw materials. The compositions and methods of formulation of the present invention are cheap, easy to carry out, and suitable for the industrial scale for commercial production.

Compounds The compounds may be in the form of the carboxylic acid and / or its salts. Salts include but are not limited to organic or inorganic salts, such as sodium salts. In addition, polyoamino acids and peptides comprising one or more of those compounds can be used. An amino acid is any carboxylic acid having at least one free amino group and includes natural and synthetic amino acids. The polyamino acids are peptides (which are two or more amino acids joined by a peptide bond) or are two or more amino acids linked by a bond formed by other groups that can be linked by, for example, an ester bond or an anhydride. The peptides may vary in length from dipeptides with two amino acids to polypeptides with several hundred amino acids. See Chambers Biological Dictionary, editor Peter M. B. Walker, Cambridge, England: Chambers Cambridge, 1989, page 215. One or more of the amino acids or peptide units can be acylated or sulphonated. Many of the compounds described herein can be derived from amino acids and can be readily prepared from amino acids by methods within the experience of those within the art based on the present disclosure and the methods described in WO96 / 30036, WO97 / 36480, US 5 / 643,957 and US 5,650,386. For example, the compounds can be prepared by reacting a single amino acid with the appropriate acylating agent or amine modifier, which reacts with a free amino moiety present in the amino acid to form amides. Protective groups can be used to avoid undesirable side reactions as is well known to those skilled in the art. With respect to protecting groups, reference is made to T.V. Greene, Protecting Groups in Organic Synthesis, Wiley, New York (1981), the description of which is incorporated herein by reference. The compound can be purified by recrystallization or by fractionation on one or more solid chromatographic supports, alone or cascaded. Suitable recrystallization solvent systems include, but are not limited to, acetonitrile, methanol, and tetrahydrofuran. The fractionation can be carried out on a suitable chromatographic support such as alumina, using mixtures of methanol / n-propanol as the mobile phase, reverse phase chromatography using mixtures of trifluoroacetic acid / acetonitrile as the mobile phase; and ion exchange chromatography using water or an appropriate buffer as the mobile phase. When the anion exchange chromatography is carried out, preferably a gradient of 0-500 mM sodium chloride is used.

Active Agents Active agents suitable for use in the present invention include biologically active agents and chemically active agents, including, but not limited to, pesticides, pharmacological agents and therapeutic agents. For example, biologically or chemically active agents suitable for use in the present invention, include, but are not limited to, proteins; polypeptides; peptides; hormones, and particularly hormones, which by themselves, do not pass (or may pass only a fraction of the dose administered) through the gastrointestinal mucosa and / or are susceptible to chemical cleavage by acids and enzymes in the tract gastrointestinal; polysaccharides, and particularly mixtures of mucopolysaccharides; carbohydrates, lipids; other organic compounds; or any combination thereof. Additional examples include, but are not limited to, the following, natural or recombinant sources, including synthetic, thereof: growth hormones, including human growth hormone (hGH), recombinant human growth hormones (rhGH), hormones of bovine growth, porcine growth hormones; hormones that release growth hormone; interferons, including α, β and β; interleukin-1; interleukin-2; insulin, including porcine, bovine, human, and recombinant human, which optionally has counterions including sodium, zinc, calcium and ammonium; insulin-like growth factor, including IGF-1; heparin, including unfractionated heparin, heparinoids, dermatans, chondroitins, low molecular weight heparin, very low molecular weight heparin and ultra low molecular weight heparin; calcitonin, including salmon, eel and human; erythropoietin, atrial naturético factor; antigens; monoclonal antibodies; somatostatin; protease inhibitors; adrenocorticotropin, gonadotropin-releasing hormone; oxytocin; luteinizing hormone-releasing hormone; follicle stimulating hormone; glucocerebrosidase; thrombopoietin; filgrastim; prostaglandins; cyclosporine, vasopressin; cromolyn sodium (cromolyn sodium or disodium); vancomycin; desferrioxamine (DFO); parathyroid hormone (PTH), including its fragments; antimicrobial agents, including antifungal agents; analogues, fragments, mimetics or derivatives modified with polyethylene glycol (PEG) of those compounds or any combination thereof.

Release systems The compositions of the present invention comprise a release agent and one or more active agents. In one embodiment, one or more of the release agent compounds, or salts of those compounds, or polyamino acids or peptides of which those compounds or salts form one or more of the units thereof, can be used as a release agent. mixing with the active agent before administration. The administration mixtures can be prepared by mixing an aqueous solution of the compound with an aqueous solution of the active ingredient, just before administration. Alternatively, the compound and the biologically and chemically active ingredient can be mixed during the manufacturing process. The solutions may optionally contain additives such as phosphate buffer salts, citric acid, glycols, or other dispersing agents. Stabilizing additives may be incorporated into the solution, preferably at a concentration ranging from about 0.1 to 20% (w / v). The release compositions of the present invention may also include one or more enzyme inhibitors. Such enzyme inhibitors include, but are not limited to, compounds such as actinonin or epiactinonin and derivatives thereof. Derivatives of the compounds are described in US 5,206,384. Other enzyme inhibitors include, but are not limited to, aprotinin (Trasylol) and Bowman-Birk inhibitor. The amount of active agent is an amount effective to fulfill the purpose of the particular active agent for the objective indication. The amount of active agent in the compositions is typically a pharmacologically, biologically, therapeutically or chemically effective amount. However, the amount may be less than that amount when the composition is used in a unit dosage form, such as a capsule, a tablet, a powder, or a liquid, because the unit dosage form may contain a plurality of compounds / compositions of biologically or chemically active agent or may contain a divided pharmacologically, biologically, therapeutically or chemically effective amount. The total effective amount can then be administered in cumulative units containing, in total, pharmacologically, biologically, therapeutically or chemically active amounts of biologically or pharmacologically active agent. The total amount of the active agent to be used can be determined by methods known to those skilled in the art. However, because the compositions can release active agents more efficiently than the above compositions, smaller amounts of biologically or chemically active agents can be administered than those used in the unit dosage forms or prior delivery systems to the subject, achieving even the same levels in blood and / or therapeutic effects. The present invention discloses compounds that release biologically and chemically active agents, particularly in oral, intranasal, sublingual, intraduodenal, subcutaneous, buccal, intracolonic, rectal, vaginal, mucosal, pulmonary, transdermal, intradermal, parenteral, intravenous, intramuscular and ocular systems, as well as the displacement through the blood-brain barrier. Unit dosage forms may also include any or a combination of excipients, diluents, disintegrants, lubricants, plasticizers, colorants, flavors, taste masking agents, sugars, sweeteners, salts and dosing vehicles, including but not limited to water, , 2-propandiol, olive oil, or any combination thereof. The compounds and compositions of the subject invention are useful for administering biologically or chemically active agents to any animals, including but not limited to birds such as chickens; mammals; such as cows, pigs, dogs, cats, primates and particularly humans; and insects. The system is particularly advantageous for releasing chemically or biologically active agents that in other circumstances would be destroyed or rendered less effective by the conditions encountered before the active agent reaches its target zone (i.e. the area in which the active agent of the release composition is to be released) and into the body of the animal to which they are administered. The compounds and compositions of the present invention are also useful in the administration of active agents, especially those that are not commonly releasable by a particular route, especially by the oral route, or those in which an improved release is desired. The release can be improved by releasing more active agent over a period of time, or in a particular period of time (such as to effect a more rapid release). After administration, the active agent present in the composition or unit dosage form is absorbed into the circulation. The bioavailability of the agent is easily evaluated by measuring a known pharmacological activity in the blood, for example an increase in blood coagulation time caused by heparin, or a decrease in circulating calcium levels caused by calcitonin. Alternatively, the circulation levels of the active agent itself can be measured directly.

DESCRIPTION OF THE PREFERRED MODALITIES The following examples illustrate the invention without limitation. All parts are given by weight unless otherwise indicated.

Example 1 - Preparation of Compound Method A: Preparation of Compound 26. A spherical bottom flask of ΔL equipped with a magnetic stirrer was charged with 2-amino-p-cresol (1.71 g, 13.88 mmol, 1 equiv.), Hydroxide 2M aqueous sodium (20 ml). Methyl azeloyl chloride (3.08 g, 13.96 mmol, 1.01 eq.) Was added dropwise to the solution, with stirring, at 0 ° C. After the addition, the reaction mixture was allowed to warm to room temperature and was stirred for 4-5 hours at room temperature. The pH of the solution was maintained at about 11-12 by the addition of 20% sodium hydroxide. The solution was then extracted with ethyl acetate (3 x 30 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was redissolved in THF (50 ml) and treated with 2N NaOH (20 ml). The resulting mixture was stirred at room temperature for 8 hours. The CCF (Thin Layer Chromatography) indicated that the reaction had been complete. The mixture was concentrated in vacuo and acidified. The resulting solid was collected, recrystallized with methanol / acetone / water to give 2.5 g of product. Compounds 11-15, 18-21, 24, 25, 27-35, 48-50, 54-56, 58, 65-68, 73, 83, 110, 120 and 131-133 were also prepared by this method using the appropriate initial materials. Compounds 120, 122 and 123 were also prepared by this method using the appropriate starting materials, except that the reaction was carried out containing THF.

Method B: Preparation of compound 60. A suspension of 1.03 g (5.62 mmoles) of 3-amino-4-fluorohydrocinnamic acid and 20 ml of methylene chloride was treated with 1.45 ml (1.24 g, 11.4 mmol) of trimethylsilyl chloride and it was heated to reflux for 150 minutes. The reaction mixture was collected at 0 ° C and treated with 2.4 ml (1.74 g, 17.2 mmol) of triethylethylamine. After stirring for 5 minutes, this mixture was poured into an addition funnel and slowly added to a 0 ° C solution of 1.21 g (6.09 mmoles) of 0-acetylsalicyloyl chloride and 10 ml of methylene chloride. The reaction mixture was heated to 25 ° C and filtered for 18 hours. Washing with 3% aqueous hydrochloric acid (2 x 20 ml), water (1 x 20 ml) and brine (1 x 15 ml), drying over sodium sulfate and concentration in vacuo gave a yellow-brown solid. The solid was recrystallized from 15 ml of 65% ethanol / water to give a brown solid. The solid was dissolved in aqueous solution saturated with sodium bicarbonate. After stirring until the acetate was cleaved (by CLAP), the solution was acidified, causing a precipitate to develop. Filtration isolation gave 0.58 g (38% yield) of the product, mp = 202-4 ° C. Compounds 59, 61 74-81, 107, 109 and 111-113 were also prepared by this method using the appropriate starting materials. Compounds 37, 42, 44 and 134 could also be prepared by this method using the appropriate starting materials.

Method C: Preparation of compound 1. 1,4-Benzodioxan-2-one (4.50 g, 30 mmol) in acetonitrile (75 ml) was dissolved in a 250 ml round bottom flask equipped with a magnetic stir bar, purge Argon and cold water condenser. Triethylamine (4.17 ml, 3.03 g, 30 mmol) and 4- (4-aminophenyl) butyric acid (5.37 g, 30 mmol) were added. The reaction mixture was refluxed for 2 h, stirred overnight at 25 ° C and concentrated in vacuo. The brown residue was taken up in methylene chloride and washed with IN aqueous hydrochloric acid solution (1 x 100 ml). The solid that formed was isolated by filtration, and rinsed with INN hydrochloric acid aqueous solution and water. The resulting solid was dried overnight in a vacuum oven, yielding the compound as a brown solid (9.19 g, 93% yield). Compound 53 was made by this method starting from dihydrocoumarin and 8-aminocaprylic acid.

Method D: Preparation of compound 36. 8- (N-3,5-dinitrosalicyloyl) aminocaprylic acid was prepared using Method T starting from 3,5-dinitrosalicylic acid and 8-aminocaprylic acid. A solution of 0.7 g (1.9 mmol) of 8- (N-3,5-dinitrosalicyloyl) aminocaprylic acid and 40 ml of ethyl acetate was treated with 70 mg of 10% Pd / C and placed under a hydrogen atmosphere. for 18 hours. The catalyst was removed by filtration. The filtration was concentrated in vacuum. The residue was recrystallized from methanol / acetone / water to give 0.4 g (62% yield) of the product, mp = 156-7 ° C. Method E: Preparation of compound 2. 4- (4- (4-benzyloxyphenoxyacetyl) amino) phenyl) butyric acid was prepared by the reaction of 4- (4-aminophenol) butyric acid with 4-benzyloxyphenoxyacetyl chloride using Method C. A suspension of 5.00 g (11.9 mmol) of 4- (4- (4-benzyloxyphenoxyacetyl) amino) phenyl) butyric acid and 150 ml of methyl alcohol was stirred under argon for about 20 minutes. A catalytic amount (0.4 g) of 10% palladium on charcoal was added in one portion. The reaction vessel was evacuated. The reaction was maintained under a nitrogen atmosphere overnight at room temperature. The palladium on charcoal was then filtered and the filtrate was concentrated under vacuum to give the product as a white solid. Compounds 8, 9 and 38 were also prepared by this method using the appropriate starting materials.

Method F: Preparation of compound 39. A suspension of 10.82 g 59.7 mmol) of 5-fluoroisatoic anhydride, 9.50 g of 8-aminocaprylic acid, 16.8 g of 50% by weight of K2C03 / water and 40 ml of dioxane was heated to reflux . 2 hours later, the reaction is complete (as measured by CLAP). The dark purple solution was cooled to 25 ° C and acidified with 3% aqueous hydrochloric acid to pH = 4.00, causing a dark precipitate to form. The solid was isolated by filtration and recrystallized from 65% ethanol / water to give 11.66 g (67% yield of the product as a yellow solid, mp = 108-9 ° C. Compounds 3-7, 10, 40 and 43 they were also prepared by this method using the appropriate initial materials.

Method G: Preparation of compound 62. A mixture at 5 ° C of 4- (4-aminophenyl) butyric acid (1.0 eq) and 6N aqueous hydrochloric acid (5.44 eq) was treated with 1.05 eq of a 3N aqueous solution of sodium nitrite. sodium, adding slowly to keep the temperature below 5 ° C. A solution of 2.8N aqueous potassium iodide (1.01 eq) was added. The reaction was stirred overnight. The layers were separated. The organic phase was purified by flash chromatography using methanol / methylene chloride as eluent to give 4- (4-iodophenyl) butyric acid. A solution of 4- (4-iodophenyl) butyric acid (0.86M) and dimethylformamide was treated with 2 eq of potassium carbonate, 1.5 eq of 2-hydroxythiophenol and a catalytic amount of cupric acetate (0.01 eq). The reaction mixture was refluxed for 18 hours, cooled to 25 ° C, acidified with aqueous acid and extracted with ethyl acetate. The organic phase was concentrated. The residue was purified by flash chromatography using ethyl acetate / hexane as eluent to give 4- (4- (2-hydroxyphenyl) thiophenyl) butyric acid. A 0.33M solution of 4- (4- (2-hydroxyphenyl) thiophenyl) butyric acid and ethyl acetate / acetic acid was treated with a 9.8M aqueous solution of hydrogen peroxide. After stirring for 12 hours, the layers were separated. The organic phase was concentrated. Purification of the residue by flash chromatography using toluene / acetone / hexane as eluent gave the product.

Method H: Preparation of compound 82. A solution at 0 ° C of 3.97 g (17.8 mmol) of 9-bromo-l-nonanol and methylene chloride was treated with a solution of 2.91 g of 2-nitrophenylisocyanate and 10 ml of chloride of methylene. The reaction mixture was heated to reflux for 2 hours, stirred at 25 ° C for 16 hours and concentrated in vacuo. The yellow solid was identified as N- (2-nitrophenyl) carbamate 9-bromononyl and was used as such. A suspension of 2.99 g (7.72 mmol) of 9-bromononyl N- (2-nitrophenyl) carbamate, 1.61 g (23.3 mmol) of sodium nitrite, 4.50 ml (4.72 g, 78.5 mmol) of acetic acid and 15 mL of dimethyl sulfoxide was stirred at 35 ° C for 7 hours. The reaction mixture was acidified with 3% aqueous hydrochloric acid and extracted with diethyl ether (3 X 20 ml). The combined organic layers were extracted with 2N aqueous sodium hydroxide solution (3 X 20 ml). The basic aqueous phases were acidified with 3% aqueous hydrochloric acid, causing a precipitate to develop. The solid was collected by filtration to give 0.79 g (yield 30%) of the compound, mp = 90-1C.

Method I: preparation of compound 64. 4- (4- (2-Aminobenzoyl) aminophenyl) butyric acid was prepared using Method F starting from isatoic anhydride and 4- (4-aminophenyl) butyric acid. A suspension of 4.73 g (16.0 mmol) of 4- (4- (2-aminobenzoyl) aminophenyl) butyric acid and 40 ml of triethyl orthoformate was placed under an argon atmosphere and refluxed for 18 hours. The reaction became clear during reflux. The reaction mixture was cooled to room temperature and the resulting solid was collected by filtration to give 4.47 g (88% yield) of the product, mp = 201-204 ° C.

Method J: Preparation of compound 63. A 0.05M solution of 4- (4- (2-hydroxyphenyl) thiophenyl) butyric acid in methylene chloride was treated with 4 eq of N-chloroperbenzoic acid at 0 ° C. The reaction mixture was allowed to warm to 25 ° C and stirred for 12 hours. The solvent was separated. The residue was purified by flash chromatography using ethyl acetate / hexane / acetic acid as eluent to give the product.

Method K: Preparation of compound 84. 8-N- (2-aminobenzoyl) aminocaprylic acid was prepared using Method F, starting from isatoic anhydride and 8-aminocaprylic acid. A suspension of 6.88 g (24.7 mmol) of 8-N- (2-aminobenzoyl) aminocaprylic acid and 100 mL of methylene chloride was cooled to 0 ° C and treated with a solution of 2.00 ml (2.08 g, 12.4 mmol) of hexamethylene diisocyanate and 5 ml of methylene chloride. The reaction mixture was refluxed for 2 hours, cooled to 25 ° C and diluted with 20 ml of ethanol. The resulting solid was isolated by filtration and recrystallized from ethyl acetate / ethanol / water, 1/2/1, yielding a total of 5.15 g (57% yield) of the product as a brown solid, mp = 138-142 ° C . The compound 135 can also be prepared by this method using the appropriate starting materials.

Method L: Preparation of compound 51. 8- (N-6-chloro-2-methoxybenzoyl) aminocaprylic acid was prepared using Method A, starting with 2-chloro-6-methoxybenzoic acid and 8-aminocaprylic acid. A suspension of 1.27 g (3.72 mmol) of 8- (N- 6-chloro-2-methoxybenzoyl) aminocaprylic acid in 200 ml of methylene chloride under an argon atmosphere was cooled to 0 ° C and treated with 8 ml of solution of boron tribromide 1.0 M in methylene chloride. After stirring for 60 minutes, the TLC (Thin Layer Chromatography) indicated that the reaction was complete. The reaction mixture was cooled with water and stirred for 30 minutes. The layers were separated. The aqueous layer was extracted with methylene chloride (2 X 30 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The resulting white solid was recrystallized from ethanol / acetone / water to give 0.5 g (43% yield) of the product, mp = 156-7 ° C.

Method M: Preparation of compound 17. A solution of 4.28 mL (0.034 mol) of dihydrocoumarin, 75 mL of acetonitrile, 4.79 mL of triethylamine (3.48 g, 0.0343 mol), and 5.62 g of 3- (4-aminophenyl) propionic acid (0.034 mol) was stirred under argon and the flask was heated at reflux for 18 hours. The reaction was cooled to room temperature, and the acetonitrile removed under vacuum. The residue was stirred in methylene chloride and IN aqueous hydrochloric acid, forming a white solid. The solid was filtered, rinsed with water, and methylene chloride, then dried under vacuum at 50 ° C to give 9.17g (86.1% yield) of the product mp = 163-165 ° C. Compound 16 was also prepared by this procedure using the appropriate starting materials.

Method N: Preparation of compound 52. A solution of 10.0 g (65.8 mmol) of 3-hydroxyphenylacetic acid in 50 ml of xylene was treated with 6.45 ml (68.4 mmol) of acetic anhydride. This mixture was refluxed for about 2.5 hours until most of the byproducts of xylene and acetic acid were distilled. The oligo- (3-hydroxyphenylacetic acid) was isolated as a brown oil. This oil was dissolved in 150 ml of 1,4-dioxane.

A solution of 9.97 g (62.7 mmol) of 8-aminocaprylic acid and 34.5 ml of 2N NaOH solution was added to the oligomer solution. The reaction mixture was heated to reflux overnight. The dioxane was then removed under vacuum. The brown residue was taken up in 2N NaOH and extracted with two 100 ml portions of ethyl acetate. The aqueous layer was then acidified with 2N sulfuric acid solution and then extracted with three 100 ml portions of ethyl acetate. The combined ethyl acetate layers were decolorized with activated charcoal, dried with sodium sulfate, and concentrated under vacuum. The resulting brown oil was then purified by column chromatography using a column of silica gel with ethyl acetate: hexane: acetic acid (60: 40: 1) as the mobile phase. The resulting white solid was washed with hot water (40-50 ° C) to give the product as a white solid. Compounds 41, 45 and 47 were also prepared by this method using the appropriate starting materials.

Method O: Preparation of compound 22. A solution of 3.0 g (11.0 mmol) of 3 - (- 4- (2-aminobenzoyl) aminophenyl) propionic acid and 10 ml of methylene chloride. Acetic anhydride (1.12 g, 1.04 ml, 11.0 mmol) was added dropwise over 30 minutes. After all the acetic anhydride was added, the reaction was stirred at room temperature for 18 hours. The reaction was complete according to what was determined by CLAP. The resulting solid was isolated by filtration. The resulting white solid was dried in a vacuum oven overnight producing the product.

Method P: Preparation of compound 23. A mixture of 5.13 g (27.9 mmol) of cyclic 2-sulfobenzoic anhydride, 5.0 g (27.9 mmol) of 4- (4-aminophenyl) butyric acid and 100 ml of acetonitrile was stirred for 18 hours . The milky solution was concentrated. The residue was taken up in 50 ml of cold aqueous hydrochloric acid, extracted with ethyl acetate (5 X 50 ml) and concentrated. The residue was purified by column chromatography using acetonitrile as eluent to give the product.

Method Q: Preparation of compound 57. A solution of 10 g of 5-chloro-2-hydroxybenzamide (58.0 mmol), pyridine (22 ml) and acetonitrile (25 ml) was stirred in an ice bath. Ethyl chloroformate (6.1 ml, 0.0638mol) was added dropwise. The pink solution was stirred for 30 minutes at < 10 ° C. The ice bath was replaced with an oil bath. The reaction mixture was heated to 95 ° C and the volatiles were distilled (43ml). The reaction was cooled to room temperature, causing a white solid to form. The mixture was poured into water (100 ml) and acidified with concentrated aqueous hydrochloric acid. The resulting solid was filtered and recrystallized from hot ethanol to give 9.77g of 6-chloro-2H-l, 3-benzoxacin-2,4 (3H) -dione.

A suspension of 9.77 of 6-chloro-2H-1, 3-benzoxacin-2,4 (3H) -dione (50 mmol) and 60 ml of DMF was treated with a solution of 1, 10-dibromodecane (52.52g, 0.175). mol) and DMF (60ml) was added. Diisopropylethylamine (9.6 ml, 55 mmol) was added dropwise. A thermometer and a condenser were attached, and the flask was placed in an oil bath. The reaction was heated at 60 ° C for about 3 hours, cooled to 47 ° C and hexanes (150ml) added. The mixture was diluted with water (175ml). The resulting solids were removed by filtration. The aqueous layer was removed with hot hexanes. The resulting solids were filtered from the hexane layer and recrystallized from the hot hexanes to give 10.39 g of 6-chloro-3- (10-bromodecyl) -2H-1,3-benzoxacin-2,4 (3H) -dione. A mixture of 6-chloro-3- (10-bromodecyl) -2H-1,3-benzoxacin-2,4 (3H) -dione (10.39g, 0.025mol), sodium nitrite (5.52g, 0.08mol) and DMSO (60ml) was stirred under argon. Acetic acid (14.9ml) was added and the reaction was heated at 75 ° C for 6 hours. After being cooled to room temperature, the reaction mixture was then dissolved in ethyl acetate and washed with 0.5 N HCl (2x) and 2N NaOH (2x). The sodium hydroxide layers were stirred for 2 hours and 20 minutes, and the solution was acidified with 2M H2SO4. The solids were then filtered and recrystallized from methylene chloride and hexanes. Performance: 3.0g.

Method R. Preparation of compound 71. A solution of 3.22 g (18.4 mmol) mono-methyl phthalate, 2.90 ml (2.11 g, 20.8 mmol) of triethylamine and 20 ml of acetone was cooled in an ice bath (with salt) and treated with a solution of 2.00 ml (2.27 g, 20.9 mmol) of ethyl chloroformate and 10 ml of acetone, added by dripping for 20 minutes. The white cloudy solution was stirred for 15 minutes and treated with a solution of 2.53 g (38.9 mmol) of sodium azide and 8 ml of water. After stirring the turbid solution for 30 minutes, it was poured into 50 ml of ice water and extracted with toluene (3 x 30 ml). The toluene phase was dried over sodium sulfate and heated to reflux for 80 minutes. The reaction mixture was cooled to 0 ° C. In several portions, 3.11 g (18.8 mmol) of 3- (4-aminophenyl) propionic acid was added over 5 minutes. After stirring for 64 hours, the reaction mixture was diluted with 80 ml of 3: 1 dioxane / water solution and heated to reflux for 3 hours. The cold mixture was extracted with 2N sodium hydroxide (1 x 30 ml) and 0.5N (2 x 30 ml). The combined aqueous layers were acidified, causing a white solid to form. Filtration isolation, dissolution in ethyl acetate, filtration of the insoluble material and concentration of the filtrate gave 1.13 g of the solid product.

Method S Preparation of compound 69. A suspension of 3.17 g (21.7 mmol) of 4-hydroxyquinazoline, 3 drops of dimethylformamide and 20.0 ml of thionyl chloride was heated to reflux. After 150 minutes, the light yellow solution was cooled to 30 ° C. The excess thionyl chloride was distilled in a 50 mm vacuum. After 15 minutes at a 1.0 mm vacuum, the white / yellow white product content of the flask was taken up in 30 ml of methylene chloride and treated with a suspension of 3.61 g (21.9 mmol) of 3- (4-aminophenyl) ) propionic and 40 ml of isopropanol. The reaction mixture was stirred for 18 hours. The light yellow solid was isolated by filtration and dried in a vacuum oven to give the product. Compound 70 was also prepared by this method except that the acidification was carried out only at pH 4.56 to isolate the free amine.

Method T Preparation of compound 46. Acetic anhydride (6.45 ml, 6.98 g, 68.4 mmol), 4-hydroxyphenylacetic acid (10.00 g, 65.8 mmol), and xylenes (50 ml) were added to a 100 ml three-necked flask equipped with a magnetic stirring bar, a thermometer and a Dean-Stark trap with condenser. The flask was heated to reflux, the reaction mixture was rinsed to a yellow solution around 100 ° C. Most of the volatile compounds (xylenes and acetic acid) were distilled from the Dean-Stark trap for 2 hours (135-146 ° C). The distillation continued for another hour, during which the temperature of the vessel slowly rose to 190 ° C and the distillate slowly began to be difficult. The residue was poured when it was still hot in an aluminum tray. After cooling, a brown waxy solid formed. 2N sodium hydroxide solution (34.4 ml, 36 g, 68.7 mmol) and 8-aminocaprylic acid (9.94 g, 62.5 mmol) were added to the solution of oligo (4-hydroxyphenylacetic acid) (11.06 g, 81.3 mmol) and dioxane (150 ml), added for 5 minutes. The reaction mixture was heated at 90 ° C for . 5 hours (time at which it was determined that the reaction had ended, by CLAP). The light orange reaction mixture was cooled to 40 ° C. The dioxane was removed in vacuo. The brown residue was taken up in 2N sodium hydroxide, extracted with ethyl acetate (2 x 100 ml), and acidified. Extraction with ethyl acetate (3 x 100 ml), decolorizing with charcoal, drying over sodium sulfate, and concentration in vacuo gave a brown oil. Trituration with hot water (2 times) gave a dark brown solid which was recrystallized twice from ethanol / water to give 2.48 g of the product as a dark brown solid.

Compounds 106 and 108 were also prepared by this method using the appropriate starting materials.

Method U. Preparation of compound 72. A suspension of 4.11 g (25.3 mmol) of 4-hydroxycoumarin, 4.54 g (25.3 mmol) of 4- (4-aminophenyl) butyric acid, and 20 ml of acetic acid was heated to reflux for 7 days. The reaction mixture was cooled to 25 ° C, causing a matt white solid to form, which was collected by filtration. The filtrate was diluted with 50 ml of water, causing a second solid to form, which was also collected by filtration. The two solids were combined and recrystallized from 65% ethanol / water to give 0.62 g of the product.

Method V. Preparation of compound 85. A suspension of 5.00 g of 10-aminodecanoic acid (26.7 mmol) in 70 ml of methylene chloride was treated with 6.78 ml of chlorotrimethylsilane (5.80 g, 53.5 mmol) and allowed to reflux for 140 hours. minutes The reaction mixture was collected at 0 ° C and then treated with 5.58 ml of triethylamine (4.1 g, 40.1 mmol). After this mixture was stirred for approximately 20 minutes, a solution of 3.91 ml of o-fluorobenzoyl chloride (4.24 g, 26.7 mmol) in 10 ml of methylene chloride was added dropwise to the reaction mixture over a period of time. 15 minutes. The reaction mixture was allowed to stir for 30 minutes, at 0 ° C and then for 18 hours at 25 ° C. The methylene chloride was removed in vacuo and 100 ml of NaOH (2N) solution was added to the residue. This mixture was allowed to stir for 1 hour before the mixture was acidified to pH = 1 with hydrochloric acid solution (2M). The acidified mixture was then extracted with ethyl acetate (2 X 100 ml), decolorized with activated charcoal, dried over sodium sulfate and concentrated in vacuo. The resulting white solid was recrystallized from a 50% ethanol / water mixture yielding a white solid, which was allowed to dry for 24 hours in vacuo at 25 ° C. The yield of the product was 6.57 g (79.5%), mp = 85-86 ° C. Compounds 86-101 were also prepared by this method by reacting the appropriate amino acid with the appropriate acid chloride.

Method W. Preparation of compound 102. A suspension of 20.72 g of glycine (0.276 mol) in 150 ml of methylene chloride was treated with 70.06 ml of chlorotrimethylsilane (59.97 g, 0.552 mol) and left to reflux for 2 hours. The reaction mixture was cooled to 0 ° C and then treated with 115.41 ml of triethylamine (83.79 g, 0.828 mol). After this mixture was stirred for approximately 20 minutes, a solution of 20.72 g (0.276 mol) of 4-methoxy-2-acetylbenzoyl chloride (58.70 g, 0.276 mol) in 75 ml of methylene chloride was added dropwise to the mixture. the reaction mixture for a period of 15 minutes. The reaction mixture was allowed to stir for 30 minutes at 0 ° C and then for 18 hours at 25 ° C. The methylene chloride was removed in vacuo and 200 ml of NaOH solution (2N) was added to the residue. This mixture was allowed to stir for several hours before the mixture was acidified to pH = 3 with hydrochloric acid solution (2M). The resulting solids were filtered and dried in vacuo at 40 ° C. The solids were recrystallized from water / ethanol (3/1) to yield a solid, which was allowed to dry for 24 hours in a row at 25 ° C. The yield of the product was 27.35 g (44%) mp = 185.5-189 ° C. The sodium salt of the above solid was produced by dissolving in 150 ml of ethanol with heating. Sodium hydroxide (4.95 g in 14.5 mL of water) was added to the ethanol solution and cooled to room temperature. The resulting solid was filtered using heptane to aid filtration and wash the solids. After drying, a dark brown solid (27.73, 92.37%) pf > 230 ° C. CHN calculated for C? 0H10? O5Na-0. 0H2O: C, 47.21; H, 4.28; N, 5.51; Na, 9.04; Found C, 47.14; H, 4.32; N, 5.36; Na, 8.45 and 2.83% water.Method X. Preparation of compound 103. A suspension of 25.0 g of β-alanine (0.281 mol) in 300 ml of methylene chloride was treated with 71.33 ml of chlorotrimethylsilane (61.06 g, 0.562 mol) and left to reflux for 1.5 hours. . The reaction mixture was cooled to 0 ° C and then treated with 117.50 ml of triethylamine (85.30 g, 0.843 mol). After this mixture was stirred for approximately 20 minutes, a solution of acetylsalicyloyl chloride (55.73g, 0.281 mol) in 150 ml of methylene chloride was added dropwise to the reaction mixture over a period of 15 minutes. The reaction mixture was allowed to stir for 30 minutes at 0 ° C and then for 18 hours at 25 ° C. The methylene chloride was removed in vacuo and 200 ml of NaOH solution (2N) was added to the residue. This mixture was allowed to stir for 1 hour before the mixture was acidified to pH = 1 with sulfuric acid (3M). The resulting oil was extracted with ethyl acetate (3 x 200 ml), dried over sodium sulfate and the solvent removed in vacuo. The solids were recrystallized from ethyl acetate / hexanes (1/1) to yield a solid, which was allowed to stir for 24 hours in vacuo at 25 ° C. The yield of the product was 9.20 g (16%). CHN calculated for C10H N? 04: C, 57.031; H, 5.27; N, 6.67 found C, 57.41; H, 5.30; N, 6.69. The sodium salt of the above solid was made by dissolving in 50 ml of ethanol with heating. Sodium hydroxide (1.79 g in 5.25 mL of water) was added to the ethanol solution and cooled to room temperature. The resulting solid was filtered. After drying, a solid (5.80 g) was obtained, mp 231-235 ° C. CHN calculated for C? 0H? 0N? O4Na-0.35H2O: C, 50.56; H, 4.54; N, 5.90; Na, 9.68; Found C, 50.30; H, 4.37; N, 5.72; Na, 9.55 and 2.68% water.

Method Y Preparation of compound 104. 4-Methoxysalicylic acid (98.74 g, 0.59 mol) in methylene chloride (500 ml) was stirred while in an ice bath.

Triethylamine (123.4 ml, 1.5eq) and acetyl chloride (46.2 ml, l.leq) were added dropwise, respectively. The solution was removed from the ice bath and stirred at room temperature for two nights. The reaction was followed by CLAP. The reaction mixture was washed with 0.5 N HCl (2x200 ml) and water (2x200 ml). The organic layer was dried over sodium sulfate and reduced in vacuo. The resulting solid was recrystallized from methylene chloride / hexanes.

The yield was 61.72 g of 4-methoxy-2-acetyl benzoic acid. The structure was confirmed by XH NMR. To the mixture of 4-methoxy-2-acetyl benzoic acid (20 g, 0.195 mol) and methylene chloride (100 ml), thionyl chloride (13.8 ml, 2 eq) and 1 drop of DMF were added. The mixture was refluxed for 1.5 hours, then cooled to room temperature and reduced in vacuo to give an oil. The 4-methoxy-2-acetyl benzoyl chloride was used without purification. A suspension of 4- (4-aminophenyl) butyric acid (7.87 g, 0.044 mol) in methylene chloride (100 ml) was treated with TMS chloride (11.2 ml) and allowed to reflux for 1.5 hours. The reaction mixture was cooled to 0 ° C and then treated with triturated triethylamine (18.4 ml). Subsequently, this mixture was stirred for approximately 20 minutes, a solution of the 4-methoxy-2-acetyl benzoyl chloride (10 g) prepared above in methylene chloride (10 ml) was added dropwise to the reaction mixture over a period of time. 15 minute period. The ice bath was removed and the mixture stirred at room temperature overnight. The methylene chloride was removed in vacuo and 2N NaOH solution (100 ml) was added to the residue. This mixture was allowed to stir for 1 hour before the mixture was acidified to pH = 1 with 2M HCl. The acidified mixture was then extracted with ethyl acetate (2x100 ml), decolorized with activated charcoal, dried over sodium sulfate, and concentrated in vacuo. The resulting white solid was crystallized from a 50% ethanol / water mixture yielding a white solid, which was allowed to dry for 24 hours in vacuo at 25 ° C. CHN calculated for C18H19N05: C, 65.54; H, 5.81; N, 4.25; Found C, 65.49; H, 5.84; N, 4.23. The yield was 7.76 g (53.6%), mp = 177-182 ° C.

Method Z Preparation of compound 105. 4-Bromobutyric acid (26.17 g, 0.16 mol) was added to methanol (150 ml) and several drops of sulfuric acid were added. This solution was refluxed for 3 1/4 hours. The TLC (1: 1 EtOAc / Hex) was performed showing the completion of ester formation. The mixture was reduced in vacuo to an oil. The oil was dissolved in methylene chloride and washed with water, saturated with sodium bicarbonate, and brine. The organic layer was dried over sodium sulfate and reduced in vacuo. The structure was confirmed by 1 H NMR and yielded 20.26 g of methyl (4-bromo) butanoate. 6-Chlorocarsalam (12.4 g, 1.12eq), methyl (4-bromo) butanoate (10.13 g, 1.0 eq) and 10.13 g of sodium carbonate (10.13 g, 1.12 eq) in 50 ml of DMA were stirred. The solution was refluxed for 4.5 hours, and then cooled to room temperature overnight. The solids were filtered and washed with ethanol, water and 2N NaOH were added to the filtrate. The mixture was heated for 2.5 hours. A CLAP was performed showing the conclusion of the hydrolysis. The solution was acidified with concentrated HCl to a pH of about 1. The resulting white solid was filtered, placed over P2O5 in vacuo overnight. The solid was recrystallized from methanol / water, filtered and dried yielding 7.35 g of the compound. CHN calculated. For CnH? 2N04Cl: C, 51.28; H, 4.69; N, 5.44; Found C, 50.92; H, 4.59; N, 5.46. Melting point of 136-140 ° C. Note that the melting points of the other batches of this compound made by the same method, showing the purest product with the elemental analysis, were in the range of 153-155 ° C.

AA method. Preparation of compound 118. Sodium carbonate (5.37 g, 0.0506 mol) was added to a 3-neck spherical bottom flask containing 6-chlorocarsalam (10. Og, 0.0506 mol) and dimethylacetamide (50 ml). Ethyl-5-bromoheptanoate (10.91g, 0.0460 mol) was added in one portion to the reaction mixture with stirring, and the reaction mixture was heated. The reaction temperature was maintained at 80 ° C and allowed to warm for 16 hours. The heating was discontinued, and the reaction mixture was allowed to cool to room temperature. The reaction mixture was filtered under vacuum and the filter cake was washed with two 20 ml portions of ethyl alcohol. Water was added to the filtrate until it was noted that an orange-brown solid had precipitated. This solid was isolated by vacuum filtration and washed first with 20 ml of ethyl alcohol and then with 20 ml of heptane. This solid was transferred to a spherical bottom flask and 200 ml of 2N NaOH was added thereto. Heating to reflux was started and continued for 1 hour. The reaction was then cooled to 25 ° C and the reaction mixture was acidified with 2N HCl solution. The white solid, which precipitated, was isolated by filtration, recrystallized from ethyl alcohol: water 30:70 and allowed to dry in vacuo overnight. 9.55g (63.0%) of the product were isolated. Melting point: 115-116 ° C. Combustion analysis:% C: 56.09 (calculated), 55.93 (found); % of H: 6.01 (calculated), 6.09 (found); % of N: 4.67 (calculated), 4.64 (found). 1U NMR analysis; (d6-DMSO): d 12.7, s, 1H (COOH); d 12.0, s, 1H (OH); d 8.88, t, 1H (NH); d 7.94, d, 1H (H ortho to the amide); d 7.42, dd, 1H (H for the amide); d 6.92, d, 1H (H ortho hydroxide); d 3.27, c, 2H (CH2 alpha to the amide); d 2.20, t, 2H (CH2 alpha to COOH); d 1.40, m, 4H (CH2 beta to the amide, CH2 beta to the COOH); d 1.30, m, 4H (remaining aliphatic CH2).

Compounds 114, 116 and 117 were also prepared by this method using the appropriate starting materials.

Method BB: Preparation of compound 121. A suspension of 2-amino-4-chlorophenol (17.88 g, 124.5 mmol), 8-ethoxy-8-oxooctanoic acid (25.19 g, 124.5 mmol), boric acid (0.385 g, 6.23 mmol) ), and 2-amino-5-methylpyridine (0.675g, 6.23 mmol) in 160 mL of dry toluene was heated to reflux (110 ° C) under nitrogen for 4 hours during which the water (2.5 mL) produced in the reaction was removed by azeotropic distillation in a separation unit. of Dean-Stark. Thin layer chromatography on silica gel (eluent: EtOAc / heptane: 1/1) revealed a complete reaction. A 2N aqueous solution of NaOH (125 mL) was added to the cold reaction mixture.250 mmol). The reaction mixture was heated to reflux for 4 hours, then cooled. The cold reaction mixture was diluted with ethyl acetate (300 mL) and water (150 mL). The aqueous layer was washed with two portions of ethyl acetate (250mL). After careful separation, the aqueous layer was cooled and acidified with 10% hydrochloric acid solution (86.20 mL, 250 mmol) to give a solid which was filtered, washed with hexane, and dried under vacuum. Trituration with dichloromethane gave the desired acid (8- (5-chloro-2-hydroxyanilino) -8-oxooctanoic acid) (22.39g, 60%) as a white matte solid: CLAP (Column: Higgins Kromasil 100 C18, water / acetonitrile / acetic acid: 950/50/1, 3mL / min, 220nm) Rt 5.38 min.; mp 123-124 ° C; X H NMR (DMSO d 6, 300 MHz) d: 1.28 (m, 4 H), 1.51 (m, 4 H), 2.19 (t, 2 H), 2.39 (t, 2 H), 6.83 (d, 1 H), 6.93 (dd) , 1H), 7.95 (d, 1H), 9.20 (s, 1H), 10.10 (s, 1H), 12.00 s broad, 1H); 13 C NMR (DMSO d6, 75 MHz) d: 24.29, 24.90, 28, 20, 33.58, 35.90, 116.50, 121.02, 122.20, 123.41, 127.74, 148.23, 171.93, 174.26. MS m / z 300 (M + 1) +. Analysis calculated for C? 4H18ClN04: C, 56.10; H, 6.05; Cl, 11.83, N, 4.67. Found: C, 56.07; H, 6.11; Cl, 11.98, N, 4.64. Compound 119 was prepared by the same method using the appropriate starting materials. The compounds 124-130 can also be prepared using this method with the appropriate starting materials.

Example 2 - Liberation of Parathyroid Hormone Dosing solutions were prepared for oral (PO) or intracolonic (IC) intake of the release agent compound and residues 1-34 of parathyroid hormone (PTH) (residues 1-38) for the solution with compound 103). A solution of the compound was made with the sodium salt of the compound or by converting the free acid to its solid salt by making a solution of the compound, stirring, adding one equivalent of sodium hydroxide (1.0 N), and diluting with water (for the solutions PO) or propylene 25% aqueous glycol (for CI). The final dosage solutions were prepared by mixing the compound solution with PTH standard solution (which typically had a concentration of 5 mg PTH / ml) and diluting to the desired volume (usually 3.0 ml). The amounts of the PTH compound doses are listed in Table 1 below. Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and given ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) 15 minutes before dosing. The rats were administered one of the dosing solutions at 1 ml / kg for the PO or 0.5 ml / kg for the IC. Serial blood samples were collected from the tail artery for serum determination of the PTH concentration. The serum PTH concentration was quantified by a PTH radioimmunoassay kit (Team # RIK 6101 from Peninsula Laboratories, Inc., San Carlos, CA). The results are illustrated in Table 1, below.

Example 3 - Heparin Release The intracolonic dosage (IC) of compositions containing a USP heparin and release agent compound in 25% aqueous propylene glycol were prepared by mixing. Any of the sodium salt of the compound was used or the free acid was converted to the sodium salt with one equivalent of sodium hydroxide (1.0 N). Typically, the compound and the heparin powders were mixed, 25% aqueous propylene glycol was added, NaOH solution was added, the content was sonic, then diluted to a volume of 3.0. The pH was checked and adjusted if necessary to pH = 7- 8. The amounts of the final compound and heparin doses are listed in Table 2 below. Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and given ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) 15 minutes before dosing. The dosing solutions were administered to rats fasting at a W dose volume of 1 ml / kg. Blood samples were collected by cardiac puncture after administration of ketamine (44 mg / kg). The activity of heparin was determined using activated partial thromboplastin time (APTT) according to the method of Henry, J.B., Clinical Diagnosis and Management by Laboratory Methods; Philadelphia, PA; W.B. Saunders (1979). The results are given in Table 2, below. twenty Example 4: Oral Examination of the Recombinant Human Growth Hormone (rhGH) Dosing solutions were prepared in oral form (PO) of the release agent compound and rhGH in phosphate buffer. A solution of the compound was made either with the sodium salt or the compound or by converting the free acid to its sodium salt by making a solution of the compound, stirring, adding one equivalent of sodium hydroxide (1.0 N), and diluting with phosphate buffer . The final dosage solutions were prepared by mixing the compound solution with a standard solution of rhGH (typically having a concentration of 15 mg rhGH / ml) and diluting to the desired volume (usually 3.0 ml). The amounts of the rhGH compound doses are listed in Table 3 below.

Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and were given ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) 15 minutes before dosing. The rats were administered 1 ml / kg of the dosing solution. Serial blood samples were collected from the tail artery for the determination of rhGH concentrations in serum. Serum rhGH concentrations were quantified by the rhGH immunoassay test kit (kit # K1F4015 from Genzyme Corporation Inc., Cambridge, MA). The results are presented in Table 3 below.

EXAMPLE 5 Release of Salmon Calcitonin (sCT) Oral dosage (PO) compositions of the salmon calcitonin release agent (sCT) compound in water were prepared by mixing. The amounts are listed in Table 4. 450 mg of the compound was added to 2.0 ml of water. Any of the sodium salt of the compound was used or the free acid was converted to the sodium salt by stirring the resulting solution and adding one equivalent of sodium hydroxide (1.0 N) and diluting with water. 90 μg of sCT was added to the solution. Then, water was added to bring the total volume to 3.0 ml. The solution had a final compound concentration of 150 mg / ml. (For compounds 118 and 123, the solutions were diluted to 6.0 ml and the dosage volume doubled). The total sCT concentration was 30 μg / ml. (For compound 123, a different amount of sCT was used to result in a final sCT dose of 100 μg / kg when dosed 2.0 ml / kg). Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and given ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) 15 minutes before dosing. The rats were administered 1 ml / kg of dosing solution (2 ml / kg) for compounds 118 and 123). Serial blood samples were collected from the tail artery. Serum sCT was determined by testing with the EIA equipment (Team # EIAS-6003 from Peninsula Laboratories, Inc., San Carlos, CA). The standard protocol of the kit was modified for compounds 104 and 105 as follows: they were incubated with 50 μl of peptide antibodies for 2 hours with shaking in the dark, the plate was washed, serum and biotinylated peptide was added and diluted with 4 ml of buffer, and it was stored overnight in the dark. The results are listed in Table 4, below.

The patents, applications, test methods and publications mentioned above are therefore incorporated herein by reference in their entirety. Many variations of the present invention will be suggested by themselves to those skilled in the art in light of the above detailed description. All those obvious variations are within the full scope of the appended claims. It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (16)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property. A compound, characterized in that it is selected from the group consisting of compounds 1-135 and salts thereof. 2. A composition, characterized in that it comprises: (A) an active agent; and (B) a compound selected from the group consisting of compounds

1-135, salts thereof, and mixtures thereof. 3. The composition according to claim 2, characterized in that the biologically active agent is selected from the group consisting of a biologically active agent, a chemically active agent, and a combination thereof. 4. The composition according to claim 3, characterized in that the biologically active agent comprises at least one protein, polypeptide, peptide, hormone, polysaccharide, mucopolysaccharide, carbohydrate or lipid. The composition according to claim 3, characterized in that the biologically active agent is selected from the group consisting of: growth hormones, human growth hormones (hGH), recombinant human growth hormones (rhGH), bovine growth hormones , porcine growth hormones, growth hormone releasing hormones, interferons, α-interferon, β-interferon, β-interferon, interleukin-1, interleukin-2, insulin, porcine insulin, bovine insulin, human insulin, and recombinant insulin human, insulin-like growth factor (IGF), IGF-1, heparin, unfractionated heparin, heparinoids, dermantanes, chondroitins, low molecular weight heparin, very low molecular weight heparin, ultra low molecular weight heparin, calcitonin, salmon calcitonin, eel calcitonin, human calcitonin; erythropoietin (EPO), atrial naturético factor; antigens; monoclonal antibodies; somatostatin; protease inhibitors; adrenocorticotropin, gonadotropin-releasing hormone; oxytocin; luteinizing hormone-releasing hormone; follicle stimulating hormone; glucocerebrosidase; thrombopoietin; filgrastim; prostaglandins; cyclosporine, vasopressin; cromolyn sodium, cromolyn sodium, cromolyn disodium; vancomycin; desferrioxamine (DFO), parathyroid hormone (PTH), fragments of PTH, antimicrobial agents, antifungal agents; analogs, fragments, mimetics and derivatives modified with polyethylene glycol "(PEG) of these compounds, and any combination thereof 6. The composition according to claim 3, characterized in that the biologically active agent comprises insulin, unfractionated heparin, heparin of low molecular weight, very low molecular weight heparin, ultra low molecular weight heparin, calcitonin, parathyroid hormone, erythropoietin, human growth hormones, or combinations thereof 7. A unit dosage form, characterized in that it comprises: ) the composition according to claim 2, and (B) (a) an excipient, (b) a diluent, (c) a disintegrant, (d) a lubricant, (e) a plasticizer, (f) a dye, (g) a dosing vehicle, or (h) any combination thereof 8. The unit dosage form according to claim 7, characterized in that the biologic agent The active compound is selected from the group consisting of a biologically active agent, a chemically active agent, and a combination thereof. 9. The unit dosage form according to claim 8, characterized in that the biologically active agent comprises at least one protein, polypeptide, peptide, hormone, polysaccharide, mucopolysaccharide, carbohydrate or lipid. 10. The dosage unit form according to claim 8, characterized in that the biologically active agent is selected from the group consisting of: growth hormones, human growth hormones (hGH), recombinant human growth hormones (rhGH), bovine growth hormones, porcine growth hormones, growth hormone releasing hormones, interferons, α-interferon, β-interferon, β-interferon, interleukin-1, interleukin-2, insulin, porcine insulin, bovine insulin, human insulin, and human recombinant insulin, insulin-like growth factor (IGF), IGF-1, heparin, unfractionated heparin, heparinoids, dermantanes, chondroitins, low molecular weight heparin, heparin of very low molecular weight, ultra-low molecular weight heparin, calcitonin, salmon calcitonin, eel calcitonin, human calcitonin; erythropoietin (EPO), atrial naturético factor; antigens; monoclonal antibodies; somatostatin; protease inhibitors; adrenocorticotropin, gonadotropin-releasing hormone; oxytocin; luteinizing hormone-releasing hormone; follicle stimulating hormone; glucocerebrosidase; thrombopoietin; filgrastim; prostaglandins; cyclosporine, vasopressin; cromolyn sodium, cromolyn sodium, cromolyn disodium; vancomycin; desferrioxamine (DFO), parathyroid hormone (PTH), fragments of PTH, antimicrobial agents, antifungal agents; analogs, fragments, mimetics and derivatives modified with polyethylene glycol (PEG) of these compounds; and any combination thereof. The unit dosage form according to claim 8, characterized in that the biologically active agent comprises insulin, unfractionated heparin, low molecular weight heparin, very low molecular weight heparin, ultra low molecular weight heparin, calcitonin, parathyroid hormone , erythropoietin, human growth hormones, or combinations thereof. 12. The unit dosage form according to claim 7, characterized in that the unit dosage form comprises a dosage vehicle comprising a tablet, a capsule, a powder, or a liquid. 13. The unit dosage form according to claim 7, characterized in that the dosing vehicle is a liquid selected from the group consisting of water, 1,

2-propanediol, ethanol, and any combinations thereof. 14. A method for administering an active agent to an animal in need of the agent, the method is characterized in that it comprises orally administering to the animal the composition according to claim 2. 15. A method for preparing a composition, characterized in that it comprises mixing: (A) at least one active agent; (B) the compound according to claim 1; and (C) optionally, a dosing vehicle. 16. A compound, characterized in that it comprises compound 105 and salts thereof. 17. A composition, characterized in that it comprises: (A) an active agent; and (B) the compound of claim 16.