KR20130081319A - Amino acid prodrugs - Google Patents

Abstract

The present invention relates to a prodrug comprising an amino acid bound to a medicament or drug having a hydroxy, amino, carboxy or acylated derivative thereon. The prodrug has the same usefulness as the medicament in which the prodrug is made, but it has improved therapeutic properties. In fact, the prodrugs of the present invention improve two or more of the therapeutic qualities as defined herein. The present invention also relates to pharmaceutical compositions containing the same.

Description

Amino acid prodrugs

The present invention relates to amino acid derivatives of pharmaceutical compounds and methods of treating certain diseases that are ameliorated by the administration of such agents and pharmaceutical compositions containing such agents. The present invention encompasses many improvements in physicochemical, biopharmaceutical and clinical efficacy in a variety of agents using amino acids such as covalent bond carriers for such agents.

The development of chemical compounds for the treatment of disorders, chronic diseases and certain diseases is becoming more difficult and expensive. The probability of success in such developments is often pessimistically low. In addition, the development time could reach ten years or even over ten years, leaving numerous patients without treatment for a longer period of time.

Even when effective pharmaceutical compounds have been developed, there are often side effects associated with their administration. These side effects may include aesthetic and pharmacokinetic bafflers that affect the efficacy of some existing pharmaceutical compounds, such as an unpleasant taste or odor of the pharmaceutical compound or composition. This can be a significant barrier to patient compliance with the regimen. Unwanted solubility properties of pharmaceutical compounds can cause difficulties in the formulation of homogeneous compositions. Other disadvantages associated with known pharmaceutical compounds include the following: low absorption of orally administered formulations; Low bioavailability of the pharmaceutical compounds in oral formulations; Lack of dose proportionality; Low stability of the pharmaceutical compounds in vitro and in vivo; Low penetration of the blood / brain barrier; Excessive first-pass metabolism when pharmaceutical compounds pass through the liver; Excessive enterohepatic recirculation; Low absorption rate; Ineffective compound release at the site of action; Excessive stimulation such as gastro-intestinal irritability and / or ulceration; Injections that cause pain in the parenteral administered pharmaceutical compounds and compositions; Excessively high dosages required for some pharmaceutical compounds and compositions, and other undesirable features. Some pharmaceutical compounds are administered to the body to produce toxic by-products with deleterious effects.

In this technology, new chemical compounds for the treatment of a wide variety of diseases are constantly being studied, and properties are being improved to overcome the disadvantages of the known pharmaceutical compounds mentioned above.

The present invention overcomes many of the problems associated with currently available drugs by making prodrugs. The concept of prodrugs is well known, and many examples of such prodrugs are documented in the literature and include antiviral agents such as statins, ACE inhibitors, and Acyclovir. There are many prodrugs available on the market, including various groups such as the like. However, the present invention uses amino acids as a moiety to prepare prodrugs. The prodrugs of the present invention have numerous advantages. For example, when amino acid prodrugs are administered by numerous routes, such as oral, intravenous (IV), rectal or other such methods, these prodrugs are converted into active drug molecules. A significant advantage of amino acid prodrugs is that they are nontoxic and are thereby digested or safely excreted into the body. This is a highly toxic group of statin-based drugs, Enalapril, Benazapril and ACE inhibitors, which are toxic as the promoiety itself, and are therefore highly toxic and thereby therapeutic of the active agent. by many prodrugs available on the market, such as numerous antibiotics such as pivoxil, isopropyl, Axetil, Silexetil and similar groups with reduced index It's quite different from the features that appear. On the other hand, as shown herein below, the amino acid prodrugs of the present invention also provide numerous advantages.

The present invention relates to a pharmaceutically active prodrug having an amino acid covalently bound to a pharmaceutical compound, which forms the amino acid prodrug and is administered in this form to a subject, such as a mammal.

Amino acids are the ideal model to be used as prodrugs because they are able to form various types of linkages between themselves and drugs. By definition, amino acids have two or more functional groups on them, amino groups (NH ₂ ) and carboxyl groups (COOH). For example, alpha-amino acids have a well known structure,

는 아미노산의 주 사슬이다. 그러므로, 예를 들어, 주 사슬에서 아미노기와 카르복실기 옆에, 측쇄(side chain)가 그 위에 작용기들을 가질 수 있다. 아미노산과 약제 사이에서 일어나는 공유 결합을 가능하게 하게 하는 것이 아미노산 부분 상의 작용기이다.
Wherein Ro is an amino acid side group or chain. As defined herein

Is the main chain of amino acids. Thus, for example, beside the amino and carboxyl groups in the main chain, a side chain may have functional groups thereon. It is the functional group on the amino acid moiety that enables covalent linkages between amino acids and drugs.

Agents or medicaments useful in the present invention contain functional groups on the compounds that allow the medicament to react with amino acids to form covalent bonds. Examples of functional groups present in the medicaments include NH ₂ , OH, COOH, or acid derivatives thereof such as esters, amides and the like.

The form of binding between the pharmaceutical compound and the amino acid may be as follows:

1) ester bonds (-CO-O-) resulting from condensation or transesterification of carboxylic acids with alcohols or phenolic hydroxyl groups, for example:

a) when the pharmaceutical compound has an aliphatic or aromatic hydroxyl group, the ester bond may be formed under the ester reaction conditions with a main chain carboxylic acid group of amino acids; or

b) when the pharmaceutical compound has an aliphatic or aromatic hydroxyl group and the amino acid has a side chain carboxylic acid group, an ester bond may be formed between the two under esterification conditions; or

c) when the pharmaceutical compound has a carboxylic acid group and the amino acid has a branched chain aliphatic or aromatic hydroxyl group, an ester bond can be formed between the two under esterification conditions; or

d) Substituents of acyloxy (e.g., alkoxy- or arylalkoxy-, aryloxy carbonyl) substituted or unsubstituted pharmaceutical compounds Ester group having (compound-O-CO-substituent), and the amino acid has a main chain carboxylic acid group, an ester bond can be formed between the two via a transesterification reaction; or

e) the pharmaceutical compound has an ester group with substituted or unsubstituted acyloxy (eg alkoxy- or arylalkoxy-, aryloxy carbonyl-) substituents (compound-O-CO-substituents) and the amino acid is When having a side chain carboxylic acid group, an ester bond may be formed between the two via a transesterification reaction; or

f) the pharmaceutical compound has an ester group with substituted or unsubstituted acyloxy (eg alkoxy- or arylalkoxy-, aryloxy carbonyl-) substituents (compound-O-CO-substituents) and the amino acid is When having a branched aliphatic or aromatic hydroxyl group, an ester bond may be formed between the two via a transesterification reaction; or

g) Alcohol and carboxylic acid moieties can be on the same molecule so that they can form intrinsic esters. For example, some compounds, such as Gabapentin, can form endogenous esters under ester forming conditions, and are also within the scope of this invention.

2) amide bonds (-CO-NH-) resulting from condensation of carboxylic acids with amines, for example:

a) when the pharmaceutical compound has an amino group and the amino acid has a main chain carboxylic acid group, the amide can be formed under amide forming conditions; or

b) where the pharmaceutical compound has an amino group and the amino acid has a side chain carboxylic acid group, an amide bond may be formed between the two under amide forming conditions; or

c) where the pharmaceutical compound has a carboxylic acid group and the amino acid has a main chain amino group, an amide bond may be formed between the two under amide forming conditions; or

d) If the pharmaceutical compound has a carboxylic acid group and the amino acid has a side chain amino group, an amide bond may be formed between the two under amide forming conditions.

Thus, the present invention relates to prodrugs formed thereby. As shown below herein, the formed prodrugs have advantages that are not recognized in relation to a medicament lacking an amino acid bound thereto. For example, it can improve bioavailability, efficacy, reduce toxicity, show greater solubility in water, and / or improve the ability of a drug to penetrate the cell membrane or through the vascular brain barrier, and stimulate gastrointestinal tract It is possible to improve the therapeutic index while showing less side effects such as.

The present invention therefore relates to a method of improving the therapeutic quality of a medicament, wherein the improvement in therapeutic quality is due to improved efficacy, improved therapeutic index, increased solubility in mammalian body fluids, improved passage to the cell membrane, improvement to the vascular brain barrier. Selected from the group consisting of reduced side effects such as increased passage, significantly reduced irritation and / or ulceration, reduced toxicity, and improved uptake for the corresponding agent administered to the patient in a non-prodrug form, Includes reacting the amino acid with the medicament to form a covalent bond between them and administering the product (here after "prodrug") to the patient. Prodrugs of the present invention have one or more improved qualities. In fact, preferably the prodrugs exhibit at least two of the improved qualities mentioned herein. Other advantages of the prodrugs include the wide availability of amino acids and the reactions that occur readily. The reaction to form the amide is generally efficient and the yield is very high, possibly at least about 70%, more preferably at least 80%, and most preferably at least about 90%.

1 is L-serine ester (F1) of (±) ibuprofen (F1), L-threonine ester (F2) of (±) ibuprofen and L-hydroxyproline ester (F3) of (±) ibuprofen (±) Antagonistic Properties of Acetylcholine-induced Struggle in Albino Mice after 1 Hour of Efficacy of Ibuprofen (ie Racemic Mixture) and Ibuprofen (S) (+) It is a schematic comparison based on.
Figure 2 shows L-serine ester of (±) ibuprofen (F1), L-threonine ester (F2) of (±) ibuprofen and L-hydroxyproline ester (F3) of (±) ibuprofen (F3), (±) ibuprofen and ( The efficacy of S) (+) ibuprofen was compared graphically based on antagonistic properties of acetylcholine induced writhing in albino rats after 3 hours of administration.
Figure 3 graphically shows the response relationship of dosage to mean clotting time (MCT) in minutes for L-serine ester of acetylsalicylic acid (Formulation 1).
FIG. 4 graphically shows the response relationship of dosage to average coagulation time in minutes for L-hydroxyproline ester of acetylsalicylic acid (Formulation 2).
FIG. 5 graphically shows the response relationship of dose to average coagulation time in minutes for L-threonine ester of acetylsalicylic acid (Formulation 3).
6 graphically shows the response relationship of dose to mean coagulation time in minutes for the control group (acetylsalicylic acid).
7 shows L-serine (ester of acetylsalicylic acid) (F. 1), L-threonine ester of acetylsalicylic acid (F. 2), L-hydroxyproline ester of acetylsalicylic acid (F. 3), and acetylsalicylic acid ( The efficacy of PC) is compared graphically as a relationship of average clotting time in minutes.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the terms "drug", "medicament", and "pharmaceutical" are used interchangeably and are the active compound administered to a patient without the binding of amino acids to it. Say In addition, as used herein, the medicament may be NH ₂ , OH, COOH or acylating derivatives thereof (eg, esters, anhydrides, amides and the like) and the like. It contains a functional group that can react with an amino acid, such as on the drug. When the agent is bound to an amino acid, the terms "amino acid prodrug" or "prodrug" of the present invention or synonyms for that agent are used.

Among the amino acids useful as a promoietie (ie, reacting with the above agents), these are those containing free amino and / or carboxylic acid groups of all conventional amino acids. Among them, some preferred embodiments have a relatively high solubility in aqueous media, for example 25 ° C., unbuffered aqueous solution, at least 100 g / L in deionized water, more preferably at least 250 g / L, and even more preferably Amino acids having at least 500 g / L. For example, glycine and proline have a solubility of about 250 g / L and 1620 g / L in deionized water at 25 ° C., respectively.

Other amino acids useful as promoters are those containing basic amino acid chains such as lysine. For example, lysine has a solubility of about 700 g / L in deionized water at 25 ° C.

Other amino acids useful as a promoiety are those containing hydroxyl side chains such as hydroxyproline, serine and threonine. For example, threonine, hydroxyproline and serine have a solubility of about 100 g / L, 369 g / L and 420 g / L in 25 ° C., deionized water, respectively.

Other preferred embodiments include amino acids having relatively low solubility in an aqueous medium, for example at 25 ° C., up to 10 g / L in deionized water, or for example up to 2 g / L, or For example, 0.6 g / L maximum. For example, the solubility of tyrosine in deionized water at 25 ° C. is about 0.5 g / L. Such prodrugs could be used to produce formulations with extended release properties because of the limited solubility of the prodrugs.

Other amino acids useful as a promoter are those containing carboxylic acid side chains such as glutamic acid and aspartic acid. Other amino acids useful as promoters are non-essential amino acids and non-naturally occurring amino acids.

The following reaction schemes represent the reactions discussed herein with regard to the reaction of hydroxyl, carboxyl and amine containing agents with various amino acids. In the schemes below, R is any agent present with less functional groups OH, COOH or NH ₂ , and R ₁ is

Wherein Ro is the side chain of the amino acids listed herein below:

Reaction Scheme A: The hydroxyl groups of the drug react with the carboxyl groups of the amino acids to form ester prodrugs.

Pharmaceutical Amino Acid Amino Acid Ester Prodrugs

Reaction Scheme B: The carboxyl group of the drug reacts with the hydroxyl group of the hydroxy amino acid where the hydroxyl group is on the side chain to form an ester prodrug.

Pharmaceutical Hydroxy Amino Acid Amino Acid Ester Prodrugs

Reaction Scheme C: The amine group of the drug reacts with the carboxyl group of the amino acid to form an amide prodrug.

Pharmaceutical Amino Acid Amino Acid Amide Prodrug

Scheme D: The carboxyl group of the drug reacts with the carboxyl group of the amino acid to form an anhydride prodrug.

Pharmaceutical Amino Acid Amino Acid Hydride Prodrug

Reaction Scheme E: The amine group of the drug reacts with the amine group of the amino acid to form an azo prodrug derivative.

Pharmaceutical Amino Acid Amino Acid Azo Prodrug

Reaction Scheme F: The carboxyl group of the drug reacts with the amine group of the amino acid to form an amide prodrug.

Pharmaceutical Amino Acid Amino Acid Amide Prodrug

In the schemes A-F, preferred amino acids used are shown below:

As used herein, the term “amino acid” refers to an organic compound having a carboxyl group (COOH) and an amino group (NH ₂ ) or theirs therein. In solution, these two end groups ionize to form a double ionized material through a wholly neutral material identified as zwitterion. The amines donate electrons to the carboxyl groups and the ends of the ions are stabilized in aqueous solutions by polar water molecules.

It is the side groups that can separate the amino acids from each other. Some amino acids, such as lysine, have amino groups on the side chains; Other amino acids have side chains containing hydroxyl groups such as threonine, serine, hydroxyproline and tyrosine; Some amino acids have carboxyl groups on the side chain, such as glutamic acid or aspartic acid. The functional groups on the side chain can also form covalent bonds with the agent, such as esters, amides and the like. When these side groups are involved in these bonds, such as hydroxyl groups. The bond will be depicted as OAA, where AA is an amino acid residue having a side chain with a hydroxyl group, but no hydroxyl group. AA by this definition refers to an amino acid having no hydroxy side groups because it participates in the reaction to form esters. In addition, when an ester is formed between the hydroxyl group of an amino acid and the OH group of the agent. The hydroxyl group on the carboxyl group forms a by-product with the hydrogen of the hydroxy group, thus the resulting product does not have an OH group on the carboxyl group, but has an acyl moiety. When the bond is expressed as C (= 0) -NHAA, this means that the amino acid is formed as an amide bond between the carboxy group of the agent and the amino group of the amino acid. However, as described, since the NH group from the amide bond is from the amino acid, AA is an amino acid without the amino group.

Preferred amino acids are naturally occurring amino acids. More preferred are those wherein the amino acids are α-amino acids. Also preferred is that the amino acids are in an L-configuration. Preferred amino acids include 20 essential amino acids. Preferred amino acids are Lysine (Lys), Leucine (Leu), Isoleucine (Ile), Glycine (Gly), Aspartic Acid (Asp), Glutamic Acid (Glu), Methionine (Met), Alanine (Ala), Valine (Val), Proline (Pro), Histidine (His), Tyrosine (Tyrosine) Tyr, Serine (Ser), Norleucine (Nor), Arginine (Arg), Phenylalanine (Phe), Tryptophan (Trp), Hydroxyproline (Hydroxyproline) (Hyp), Homoserine (Hsr), Carnitine (Car), Ornithine (Ort), Canavanine (Cav), Asparagine (Asn), Glutamine (Gln), Carnosine (Can), Taurine (Tau), djenkolic Acid (Djk), gamma-aminobutyric Acid (GABA), cysteine ( Cysteine) (Cys), Cystine (Dcy), Sarcosine (Sar), Threonine (Trenine) (Thr) and the like. Even more preferred amino acids are 20 essential amino acids. Lys, Leu, Ile, Gly, Asp, Glu, Met, Ala, Val, Pro, His, Tyr, Thr, Arg, Phe, Trp, Gln, Asn, Cys and Ser.

Prodrugs are prepared from a medicament having its group capable of reacting with amino acids.

Preferred agents that react with amino acids according to various schemes are as follows:

Reaction plans

Pharmaceutical A B C D E F

Cyclosporins YES

Lopinavir YES YES YES

Ritonavir YES YES YES

Cefdinir YES YES YES YES YES

Zileuton YES YES YES

Nelfinavir YES YES YES

Flavoxate YES YES YES

Candisarten YES YES YES YES YES

Propofol YES

Nisoldipine YES YES YES YES YES

Amlodipine YES YES YES YES YES

Ciprofloxacin YES YES YES YES

Ofloxacin YES YES YES YES

Fosinopril YES YES YES

Enalapril YES YES YES

Ramipril YES YES YES

Benazepril YES YES YES

Moexipril YES YES YES

Trandoraapril YES YES YES

Cromolyn YES YES YES YES

Amoxicillin YES YES YES YES YES YES

Cefuroxime YES YES YES YES YES YES

Ceftazimide YES YES YES YES YES YES

Cefpodoxime YES YES YES YES YES YES

Atovaquone YES

Gancyclovir YES YES YES

Penciclovir YES YES YES

Famciclovir YES YES YES

Acyclovir YES YES YES

Niacin YES YES YES

Bexarotene YES YES YES

Propoxyphene YES

Salsalate YES YES YES YES

Acetaminophen YES

Ibuprofen YES YES YES

Lovastatin YES YES YES YES

Simavastatin YES YES YES YES

Atorvastatin YES YES YES YES

Pravastatin YES YES YES YES

Fluvastatin YES YES YES YES

Nadolol YES

Valsartan YES YES YES

Methylphenidate YES YES YES YES

Sulfa Drugs YES YES

Sulfasalazine YES

Methylprednisolone YES

(Methylprednisolone)

Medroxyprogesterone YES

(Medroxyprogesterone)

Estramustine YES

Miglitol YES

Mefloquine YES YES

Capacitabine YES

Danazol YES

Eprosartan YES YES YES

Divalproex YES YES YES

Fenofibrate YES YES YES

Gabapentin * YES YES YES YES YES

Omeprazole YES

Lansoprazole YES

Megestrol YES

Metformin YES

Tazorotene YES YES YES

Sumittriptan YES

Naratriptan YES

Zolmitriptan YES

Aspirin YES YES YES

Olmesartan YES YES YES

Sirolimus YES

Tacrolimus YES

Clopidogrel YES YES YES

Amphotericin B YES YES YES YES

Tenenofovir YES

Unoprostone YES YES YES

Fulvestrant YES

Cefditoren YES YES YES

Efavirenz YES

Eplerenone YES YES YES

Treprostinil YES YES YES YES

Adefovir YES

The prodrugs of the present invention contain amino groups, which are basic in nature. The amino groups have various inorganic and organic acids and can form pharmaceutically acceptable salts with a wide variety. These acids which can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are non-toxic acid addition salts, ie hydrochloride, hydrobromide, hydroiodide, nitride , Sulfate, bisulfate, phosphate, formate, acetate, citrate, tartate, lactate and the like To form salts containing pharmaceutically acceptable anions.

As shown herein, in one embodiment, the present invention relates to a prodrug, wherein the prodrug is cyclosporine, and MeBmt (xy = CH = CH) or dihydro MeBmt (xy = CH). ₂ CH ₂ ) moiety, such as an esterified amino acid. Amino acids are bound to cyclosporin and other agents by covalent bonds.

The compounds of the present invention are prepared by techniques already known in the art. For example, if the medicament contains an OH group, it is referred to as cyclosporine, which is an amino acid such as acid halide or its acylated derivatives, for example amino acid fluoride, amino acid chloride ( amino acid chloride), or amino acid alkyl esters containing 1 to 6 carbon atoms, which react with the carboxy group of the drug. For example, cyclosporin under esterification conditions. Preferably, the reaction is carried out in the presence of acids such as hydrochloric acid, hydrobromic acid, para-toluenesulfonic acid and the like. Alternatively, as described herein, if a medicament has an amino group on the medicament, then the amino acid will react with the medicament under amide forming conditions to form an amide as a covalent bond, or the medicament on the medicament If it has a carboxyl group or acylated derivative, it will react with the amino group of that amino acid under amide forming conditions to form an amide bond between that amino acid and the agent. Additionally, if a medicament has a carboxyl group in the medicament, the hydroxy group of the side chain of the amino acid will react with the carboxyl group or acylated derivative, and the reaction may be used to form an ester bond between the amino acid and the medicament, as described herein. Under esterification conditions.

If an amino acid has its group reactive under reaction conditions, it is protected by a protecting group well known in the art. After the reaction is completed, the protecting group is removed. Examples of protecting groups that can be used are described in a book entitled “Protective Group in Organic Synthesis” by Theodora W. Greene, John Wiley & Sons (1981), see Is cited.

For example, if amino acids having carboxylic acid groups in their side chains, such as aspartic acid and glutamic acid, are used in the aforementioned synthesis, they will generally require protection of the side chain carboxylic acids. Suitable protecting groups include esters (eg cyclohexyl esters, t-butyl esters, benzyl esters, allyl esters) or adamantyl groups (eg 9-fluorophenyl-methyl groups Or 1- or 2-adamantyl) which can be protected after the esterification is completed, using techniques known to those skilled in the art.

Amino acids having hydroxyl groups in the side chain, such as serine, threonine, hydroxyproline and the like, and amino acids having phenolic groups in the side chain, such as tyrosine and the like, are mentioned above. If used, the amino acids will preferably require protection of the chain hydroxyl or phenolic groups. Suitable as protecting groups for hydroxyl side chain groups may be ethers such as benzyl ether or t-butyl ether. Removal of benzyl ether can be effected by liquid hydrogen fluoride, while t-butyl ether can be removed by treatment with trifluoroacetic acid. Suitable as protecting groups for phenolic side chains are benzyl ether or t-butyl ether 2,6-dichlorobenzyl, 2-bromobenzyloxycarbonyl, 2,4-di The ethers mentioned above, including nitrophenyl (2,4-dintrophenyl) and the like.

In addition, the product is substantially pure in its state by techniques known to those skilled in the art, such as chromatography, such as high pressure liquid chromatography columns (HPLC), crystallization and the like. Can be purified to become. In practice, "pure" means that the product contains only about 10% of impurities, at most in them.

Prodrugs can be made of pharmaceutical compositions, including: Prodrugs, or pharmaceutically acceptable salts, pharmaceutically acceptable solvents, esters, enantiomers, diastereomers, N-Oxides, homogeneous Polymorphs, among them, together with a pharmaceutically acceptable carrier, and alternatively but preferably with pharmaceutically acceptable excipients, as described herein. In this technique, techniques known to those skilled in the art are used.

Prodrugs used in the methods of the invention were used in therapeutically effective amounts.

The physician will determine the most appropriate dosage of the prodrugs of the invention, which will vary depending on the dosage form and the particular compound selected, and further include the patient being treated or the age of the patient, the severity of the disease being treated and the like. It will vary depending upon the identification of the various factors and prodrugs administered, but not limited to one. Doctors generally want to start treatment at a dose that is actually less than the proper dose of the compound, and increase the dose from a small amount until a reasonable effect is reached in that situation. In general, when the composition is administered orally, higher amounts of active agent will be required to produce the same effect by the smaller amount given parenterally. The compounds are useful in the same manner as the medicaments corresponding to the non-prolong form and the dosage is subject to the same dosage rules as when generally used with these other therapeutic agents. When administered parenterally, compounds are generally administered at a dose of, for example, from about 0.001 to about 10,000 mg / kg / day, and also depending on the host and the severity of the disease being treated and the compound used. Is determined.

The compounds used in one preferred embodiment are administered at doses ranging from about 0.01 mg to about 1000 mg per kilogram of body weight per day, depending on the particular mammalian host being treated or the disease, more preferably body weight per day It is in the range of about 0.1 to 500 mg / kg sugar. This dosing regimen may be adjusted by the physician to provide the optimum therapeutic response. For example, several divided doses may be administered daily, or the dose may be indicated and proportionally reduced, depending on the urgent need of the therapeutic situation.

Prodrugs may be administered by convenient methods such as oral, intravenous, intramuscular or subcutaneous infusion routes.

Prodrugs can be administered orally, eg, using an inert diluent or a digestible edible carrier, sutured in hard or soft shell gelatin capsules, or in a tablet. It can be administered by pressing or by incorporating it directly into the food to be consumed. For oral therapeutic administration, prodrugs can be incorporated with excipients and consumed tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, Used in the form of wafers, and the like. Such compositions and formulations should contain at least 1% prodrug. The percentage of compositions and formulations may, of course, vary and may conveniently be between about 5% and about 80% of the unit weight. As such the amount of prodrug used in such therapeutic compositions can be obtained in appropriate dosages. According to the present invention preferred compositions or formulations contain from about 200 mg to about 4000 mg of prodrug. Tablets, troches, pills, capsules, and the like, also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; Excipients such as dicalcium phosphate; Disintegrating agents such as corn starch, potato starch, alginic acid and the like; Lubricants such as magnesium stearate; And sweetening agents such as sucrose, lactose or saccharin may be added or flavoring agents such as peppermint, oil of wintergreen, or cherry flavor. May be When the dosage unit form is a capsule, it may comprise a liquid carrier in addition to this type.

Various other kinds of materials may be present as coatings or otherwise alter the physical form of the dosage unit. For example, tablets, pills or capsules may be coated with shellac, sugar or both. Syrups or elixirs may contain active compounds, sugars as sweeteners, methyl and propylparabens as dyes, dyes and flavoring agents such as cherry or orange flavors. Of course, the materials used to prepare the dosage unit form must be pharmaceutically acceptable and practically nontoxic at the dosages used. In addition, the active compound will be incorporated into sustained-release preparations and formulations. For example, the active ingredient is templated, which is bound to an ion exchange resin, which can alternatively be coated with a diffusion barrier coating to alter the release properties of the resin, or present The prodrugs of the invention are associated with delayed release polymers well known in the art such as hydroxypropylmethylcellulose and the like.

Prodrugs may also be administered parenterally or intraperitoneally. It is particularly advantageous to formulate parenteral compositions in dosage units for ease of administration and uniformity of dosage. Dispersions can also be prepared in glycerol (eg glycerol such as PEG 100, PEG 200, PEG 300, PEG 400 and the like), liquid polyethylene glycols and mixtures thereof and on oils. Under conditions of normal storage and use, these formulations contain a preservative to prevent the growth of microorganisms.

Pharmaceutical forms suitable for injectable use include sterile aqueous solutions (if soluble in water) or dispersions and sterile powders for the temporary formulation of the sterile injection solution or dispersion. In all cases, the formulation is usually sterile and must be fluid to the extent that injectability exists. The formulation should be stable under the conditions of production and storage and usually should be preserved against the contaminating activity of microorganisms such as bacteria and fungi. Carriers include, for example, water, ethanol, polyols (eg, glycerol, propylene glycol, and one or more liquid polyethylene glycols as disclosed herein, and the like, for example), suitable mixtures thereof, and vegetable It may be a solvent or a dispersion medium containing oils. Proper fluidity can be maintained, for example by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. Prevention of microbial activity can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. It may be possible. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Delayed absorption of injectable compositions may be possible by use in compositions of agents that delay absorption such as aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the prodrug in the required dose in the appropriate solvent with the other contents listed above, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various bactericidal active ingredients into a sterile vehicle that contains a basic dispersion medium and the required other contents from those enumerated above. In the case of sterile powders, the solutions are vacuum dried or lyophilized as necessary.

Prodrugs can also be applied topically, such as, for example, via a patch, using techniques known to those of ordinary skill in the art. Prodrugs can be administered orally by making suitable formulations of the prodrugs of the invention and using processes well known to those of ordinary skill in the art. These formulations are prepared from suitable nontoxic pharmaceutically acceptable ingredients. These ingredients are known to those of ordinary skill in the art of making oral dosage forms. Some of these ingredients can be found in Remington's Pharmaceutical Sciences, 17th edition, 1985, which is a standard reference in this field. The choice of a suitable carrier depends largely on the exact nature of the desired oral dosage form, for example tablets, lozenges, gels, patches, and the like. All of these oral dosage forms are templated within the scope of the present invention and formulated by conventional methods.

Formulations of pharmaceutical compositions will be prepared using conventional methods using one or more physicochemical and / or pharmaceutically acceptable carriers or excipients. Thus, the compounds and their pharmaceutically acceptable salts and solvates may be subjected to inhalation or insufflation (through the mouth or through the nose) or oral, oral, parenteral, or rectal administration. Will be formulated for administration. For oral administration, the pharmaceutical compositions may contain binders (e.g., pregelatinized maize starch, polyvinylpyrrolidone, or hydroxypropylmethyl cellulose; fillers) ) (E.g. lactose, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g. magnesium stearate, talc, or silica) Pharmaceutically acceptable excipients such as disintegrants (e.g. potato starch, or sodium starch glycolate) or wetting agents (e.g. sodium lauryl sulfates); It will take formulations prepared by conventional methods, for example tablets or capsules, which will be coated by methods well known in the art. The.

Liquid formulations for oral administration will take formulations such as, for example, solutions, syrups, or suspensions, or the formulations may be presented as a dried product for systems using water or other suitable vehicle before use. . Such liquid formulations include suspending agents (e.g. sorbitol syrup, corn syrup, cellulose derivatives or hydrogenated edible oils and fats; emulsifying agents) (e.g. lecithin) Or acacia) non-aqueous vehicles (e.g. almond oil, oily esters, ethyl alcohol or fractionally distilled plant oils; and preservatives (e.g. methyl or propyl para-hydride) Pharmaceutically acceptable additives such as oxybenzoate or sorbic acid), and the formulations will also contain buffering salts, flavoring agents, coloring agents and sweetening agents as appropriate. The formulations will be formulated appropriately so that an effective prodrug can have controlled release.

The prodrugs of the present invention will be formulated for parenteral administration by injection, for example by bolus injection or continuous infusion. Formulations for use as injectables will be presented in unit dosage form, eg, with a preservative added in ampoules or in multidose containers, the compositions comprising suspensions, solutions Or formulations such as emulsions in oily or aqueous vehicles and will include formulations such as suspending agents, stabilizers and / or dispersing agents. Alternatively, the prodrug will be in powder form for systems using a suitable vehicle, eg, sterile, pyrogen-free water, before use.

The prodrugs of the present invention will also be formulated with rectal infusion compositions such as suppositories or retention enemas, for example conventional suppository bases such as cocoa butter or other glycerides. It contains.

In addition to the formulations previously described, the prodrugs of the invention will also be formulated as a depot preparation. Such long active formulations will be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, prodrugs are suitable polymeric or hydrophobic materials (e.g., emulsions of acceptable oils) or ion exchange resins or poorly soluble derivatives, for example in poorly soluble salts. Will be formulated.

Pharmaceutical compositions containing the prodrugs of the present invention, if desired, will be presented as a pack or distribution device containing one or more unit dosage forms containing the active ingredients. The pack comprises a metal or plastic foil, for example a transparent plastic pack. The pack or distribution device will be accompanied by instructions for administration.

In tablet form, it is desirable to include a lubricant that facilitates the process of preparing the dosage units; Lubricants will also optimize the erosion rate and drug dissolution. If lubricant is present, it will be present in the definition of 0.01 wt% to about 2 wt%, preferably about 0.01 wt% to 0.5 wt% of the dosage unit. Suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, sodium stearylfumarate, talc, hydrogenated vegetable oils and polyethylene glycols. However, as contemplated by those skilled in the art, changing the particle size of the components and the concentration of the units in the dosage unit has a similar effect—ie improved productivity, and the rate of erosion without addition of lubricant and It can provide optimization of drug flux.

Other ingredients may also be incorporated into the dosage unit as well. Such optional ingredients to be added include, for example, one or more disintegrants, diluents, binders, enhancers or the like. Examples of disintegrants that can be used are cross-linked polyvinylpyrrolidones, for example crospovidone (for example obtained from Polyplasdonet® XL, GAF), Intercrossed carboxymethylcelluloses, for example ascroscanmelose (for example Ac-di-sol®, available from FMC), alginic acid, and sodium carboxymethyl starch (Eg, available from Xplotab®, Edward Medell Co., Inc.), agar bentonite, and alginic acid. Suitable diluents are those typically used in pharmaceutical formulations using compression techniques, for example dicalcium phosphate dihydrate (for example available from Di-Tabs®, Stauffer). Sugars obtained by crystallization with dextrin (for example, co-crystallized sucrose and dextrin such as Di-Pak®, available from Amstar), calcium phosphate, cellulose, kaolin, Mannitol, sodium chloride, dried starch, powdered sugar and the like. If used, binders are those that enhance adhesion. Examples of such binders include, but are not limited to, starches, gelatin, and sugars such as sucrose, dextrose, molasses, and lactose. Permeation enhancers may also be included in new dosage units to increase the rate of passage of an effective agent through the oral mucosa. Examples of penetration enhancers include dimethelsulfoxide ("DMSO"), dimethyl formaldehyde ("DMF"), N, N-dimethylacetamide ("DMA"), decylmethylsulfoxide ("C ₁₀ MSO). "), Polyethylene glycolmonolaurate (" PEGML "), glycerol monolaurate, lecithin, 1-substituted azacycloheptan-2-ones, in particular 1-n 1-n-dodecylcyclazacycloheptan-2-one (available under the trademark Azone.RTM. Of Nelson Research & Development, Irvine, Calif.), Lower alkanols (E.g. ethanol), SEPA (commercially available from Macrochem, Lexington, Mass.), Bile acid (cholic acid), taurocholic acid, bile salt type enhancers and surfactants (e.g. , Tergitol®, Nonoxynol-9® and TWEEN-80®).

Flavoring agents may alternatively be included in the various pharmaceutical formulations. Suitable flavoring agents may be used, for example, artificial sweeteners such as mannitol, lactose or aspartame. Although such agents are not required, colorants can also be added. Examples of colorants include water soluble FD & C dyes, mixtures of the same, or pigments corresponding thereto.

Additionally, if desired, the dosage units of the present invention may be used in combination with one or more preservatives or bacteriostatic agents such as methyl hydroxybenzoate, propyl hydroxybenzoate, chlorocresol, benzalkonium chloride. (benzalkonium chloride) and the like.

As used herein, "pharmaceutically acceptable carrier" refers to absorption delay agents for any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and pharmaceutically effective materials well known in the art. Include them. Except insofar as any conventional media or agent is incompatible with the prodrug. The use of the carrier in a therapeutic composition is templated. Supplementary active ingredients can also be incorporated into the compositions.

Dosage unit form as used herein refers to physically discrete units adapted as unitary administration to the subjects to be treated; Each unit containing a predetermined amount of prodrug is associated with the required pharmaceutical carrier to yield the desired therapeutic effect.

As described herein, prodrugs are synthesized for convenient and effective administration with the appropriate pharmaceutically acceptable carrier in dosage unit form. Unit dosages contain the main active compound, for example, in doses ranging from about 10 mg, or as low as 1 mg (for small animals) to about 2000 mg, in humans. Once in solution, the concentration of the prodrugs preferably ranges from about 10 mg / mL to about 250 mg / mL. In the case of compositions containing supplementary active ingredients, the dosage is determined by reference to the amounts and methods of use of the ingredients already mentioned. In the case of oral administration, the prodrugs will preferably be in an oral unit dosage form in which the doses are in a range of about 10 to about 50 mg.

The prodrugs of the present invention are effective when treating a disease or condition in which the corresponding medicament (without the amino acid prodrug of the present invention) is normally used.

As used herein, the term “treating” refers to reversing, alleviating or inhibiting the development of a disease, dysfunction or disease, or the development of one or more symptoms of such a disease, dysfunction or disease. Say. In addition, as used herein, the term “treatment” refers to a disease, dysfunction, or disease that can occur in a mammal, as compared to an untreated control population, or to the same mammal prior to being treated. It can be said to reduce the probability or case that there is. For example, as used herein, the term “treatment” refers to preventing a disease, dysfunction or disease, and delaying or preventing the onset of a disease, dysfunction or disease, or a disease, function And delaying or preventing symptoms associated with the disorder or disease. In addition, as used herein, the term “treatment” may be directed to reducing a disease, dysfunction or disease, or symptoms associated with a disease, dysfunction or disease seen before a mammal suffers from the disease, dysfunction or disease. have. Preventing or reducing the severity of a disease, dysfunction or disease prior to such illness is directed to administering a composition of the present invention, as described herein, wherein at the time of administration blood is not suffering from the disease, dysfunction or disease. It is about a sample. In addition, as used herein, the term “treatment” may be directed to preventing a recurrence of a disease, dysfunction or disease or recurrence of one or more diseases, dysfunctions or symptoms associated with a disease. The terms "treatment" and "therapeutically" as used herein refer to the act of "treating" as defined above.

As used herein, the term “patient” or “subject” refers to a warm animal, preferably a mammal, and includes, for example, humans, including cats, dogs, horses, cattle, There are pigs, mice, rats and primates. Preferred patients are humans.

The prodrugs of the present invention show the same utility as the corresponding agents without amino acid bonds. The prodrugs exhibit improved treatment quality. It is that the prodrugs exhibit at least one or more, preferably at least two or more, improved treatment qualities compared to a medicament that has not been modified with the prodrug of the invention prior to administration. this is

a. Improved taste and smell

b. Desired octanol / water partition coefficient (ie, solubility in water / fat), including but not limited to.

Various amino acids have different solubilities in aqueous solutions. By selecting specific amino acids, the octanol / water partition coefficient can be influenced. For example, many of the drugs in the following list are very hydrophobic. Amino acids are very hydrophilic. For example, suppose Propofol is a drug and lysine is an amino acid. Propofol is completely insoluble in water, while lysine is soluble to show 700 mg / ml solubility. When these two different molecules are esterified via ester linkage, the resulting lysine ester of propofol has a solubility of more than 250 mg / ml in water.

On the other hand, cromolyn sodium is very water soluble. For all practical purposes it is not absorbed when administered orally. By affecting its solubility in water, the absorbency can be improved. In this case, one would look for conditions that are opposite to the solubility of propofol, that is to reduce the solubility in water. By selecting amino acids with moderately low solubility in water, such as tyrosine, a suitable hydrophilic / hydrophobic balance can be obtained.

c. Improved stability in vitro and in vivo

d. Improved penetration of the blood vessel-brain barrier

e. Elimination of the first-pass effect in the liver, ie the drug is not metabolized in the liver and thus more agent is present in the body circulatory system

f. Reduction of intestinal recirculation (this improves bioavailability)

g. Painless Injections in Parenteral Formulations

h. Improved bioavailability

i. Improved change in absorption (increase vs lack in absorption)

j. Reduced side effects

k. Dose proportionality

The need for proportionality of dosage requires that when the agent is administered with increasing dose, a proportionally rising dose of the active agent is delivered into the bloodstream. This is measured by administering the drug by a route other than intravenous and determining the area under the plasma concentration vs time curve obtained after the same measurement in plasma / blood. A simple mathematical procedure is as follows: For example, a medicament is administered orally to a patient with three different doses, 10, 100 and 1000 mg, with an area under the plasma concentration time curve (AUC). Is measured. Each total AUC is then divided by dose. And the result should be the same for all three doses. If so, there is a dose proportionality. Absence of dose proportionality indicates that one or more pharmacokinetic / pharmaceutical mechanisms have been saturated and includes the number of receptor sites available for absorption, metabolism or pharmacological response.

For example, suppose in the above study, AUC values of 100, 1000 and 10,000 were obtained. In this case, dose proportionality is inadequate. In the absence of dose proportionality, the drug in the plasma is more or less in quantity and depends on what mechanism may be saturated. What follows is the possibilities: saturable absorption. If this is the case, as the dose is increased, proportionally less drug is absorbed, so the overall AUC will decrease as the dose is increased.

Saturable metabolism of elimination. Thus, in that case, the drug will circulate more and more in the blood, and the AUC will increase with increased dosage.

Saturable Pharmacological Receptor Sites: In this case, any additional agent will not increase the response because all receptor sites will someday be occupied by the agent. Thus increasing doses will not result in increased response.

Dose proportionality is a good response profile because anyone can accurately predict the pharmacological response and therapeutic effect at all doses. Dose proportionality is the desired quality for all agents. Furthermore, achieving dose proportionality is also determined by the formulation and the fed / fasted differences.

l. Selective Hydrolysis of Prodrugs at the Site of Action

m. Controlled release properties

n. Targeted Drug Delivery

o. Reduced toxicity, thereby improving treatment rate

p. Reduced dosage

q. Alteration of metabolic pathways to deliver more agents at the site of action

r. Increased Solubility in Aqueous Solutions

p. Enhanced efficacy

Thus, various dosage forms have been made possible with amino acid prodrugs and they are prepared by conventional methods:

i. Oral liquid administration (including controlled release and instant release liquids and chewable tablets containing sugar and sugar free, dye and dye free, alcohol and alcohol free formulations)

Ii. Solid oral administration (controlled and immediate release tablets, capsules, caplets)

Iii. Intravenous (injectables, ready-to-use and lyophilized powders)

Iv. Intramuscular (injectables, ready-to-use and lyophilized powders)

v. Subcutaneous (injectables, ready-to-use and lyophilized powders)

피 .Transdermal (primarily a patch)

Ⅶ. Nasal (Sprays, formulations for treatments with nebulizer)

Ⅷ. Topical (creams, ointments)

Iii. Rectum (creams, ointments, suppositories)

x. Intravaginal (creams, ointments, and pessaries)

xi. Eye drops (Eye Drops and Eye Drops)

xii. Intraoral (chewable and currently chewable tablets)

Many of the agents discussed herein, particularly in the following table, are characteristically very hydrophobic so that they settle easily even with very little water, for example. This is the case when it comes into contact with the human body (eg gastric juice). Therefore, such drugs are extremely difficult to provide, for example. There are oral formulations that are acceptable to the patient in terms of form and taste, which are drugs that can be administered as a basic daily act to provide a stable, appropriate and controlled patient dosage on storage.

Presented liquid formulations, for example. Oral administration of many of the agents shown in the table heretofore has been based primarily on the use of ethanol and oils or similar excipients as carrier media. Thus, current commercially available drink-solutions of many drugs are for example. Ethanol and olive or corn oil are used as carrier media in conjunction with solvent systems comprising ethanol and LABRIFIL as the carrier medium and equivalent excipients. For example, commercially available cyclosporin drink solutions use ethanol and olive or corn oil as carrier media in connection with Labroid as a surfactant. See, for example, US Pat. No. 4,388, 307. The use of compositions similar to the drink solutions presented in this technique, however, involves various difficulties.

Furthermore, the good taste of known oil based systems has proved problematic. The taste of known drink-solutions of some drugs is particularly unpleasant. Mixing into a moderately flavored drink, such as a chocolate drink formulation, is generally done at high dilution prior to dosing to allow for normal treatment to an acceptable level. The use of oil-based systems also requires the use of high ethanol concentrations, which are themselves inherently undesirable, particularly where pediatric administration is expected. Also, evaporation of ethanol is for example. From capsules (used in many parts, to solve the problem of good taste, as discussed or in other formulations (eg when opened)), progress is made in drug settling. When such compositions are presented, for example, in soft gelatin encapsulation formulations, this particular difficulty requires the packaging of hermetic ingredients, for example hermetic transparent packs or aluminum foil blister packs, encapsulated products. This in turn makes the goods huge, producing more expensive production costs. The storage characteristics of the formulations mentioned above are also far from ideal yet.

The level of bioavailability obtained using existing oral administration systems for many of the agents described herein is also low, and vary widely at different times during treatment, between individuals, types of individual patients, and even in single individuals. Indicates. Reports in the literature show that current treatments using commercially available pharmaceutical drink solutions provide only about 10-30% mean abslolute bioavailability, among populations. There is a noticeable deviation of. For example, deviations between humans with liver (relatively low bioavailability) and bone marrow (relatively high bioavailability) transplants. Deviations in reported bioavailability among subjects vary from one or a few percent of some patients to 90% of other patients. As already stated, noticeable changes in individual bioavailability with time are frequently observed. Thus, there is a need for consistent and high bioavailability of many of the agents shown here in patients.

The use of such dosage forms is also characterized by extreme deviations in the dosage of the patient as needed. In order to carry out an effective treatment, the drug blood concentration or the serum serum concentration must be maintained within a certain range. This required range may vary in sequence, depending on the particular disease being treated, for example whether the therapy can prevent the pharmacological activity of one or more specific agents and alternative therapies may be used as ancillary to the principle therapy. It is time. Because of the wide variation in bioavailability obtained with existing dosage forms, the daily dosages required to achieve the required serum concentrations will vary considerably between individuals and even for a single individual. For this reason, it will be necessary to monitor the blood / serum concentrations of patients undergoing medication at regular and frequent intervals. Monitoring of blood / serum concentrations should be carried out on a regular basis. This is inevitably time consuming and inconvenient and actually adds to the overall cost of the therapy.

In addition, the blood / serum concentrations of many of the agents described herein obtained using applicable dosing systems may show extreme deviations between peak and trough concentrations. It is for each patient and the effective drug concentration in the blood varies widely between each administration during individual dosing.

There is also a need to provide many of the agents described herein, particularly beta-lactum antibiotics, cyclosporin, cephalosporins, steroids, and quinolone antibiotics. And cyclosporin in water-soluble formulation for injection. Cremaphore L (CreL), used in the current formulations of many of the agents described below, is a polyoxyethylated derivative of castor oil and a toxic carrier . There have been many instances of anaphylaxis because of the castor oil component. At present, there is no formulation that allows many of these agents to reach the required concentrations in aqueous solutions because of their low solubility in water.

Beyond all such very obvious practical difficulties, there is the occurrence of unwanted additional reactions already mentioned, which are observed using the applicable oral dosage forms.

Several proposals to address these various problems have been presented in the art, including oral dosage forms of both solid and liquid. However, the primary difficulty still remaining here is that some of the agents shown in the table below are insoluble in aqueous media inherently, which prevents the use of dosage forms that may contain agents in sufficiently high concentrations. The formulations are convenient to use but yet still meet the required criteria in terms of bioavailability, for example, to achieve effective absorption from the gastrointestinal or intestinal lumen consistent with adequate and adequately high blood / serum concentrations. Formulation.

Particular difficulties faced with oral administration of these agents are inevitably limiting the use of certain drug therapies to treat conditions of disease that may be relatively less severe or dangerous. For example, when taking cyclosporin as an experimental drug, a particular area of difficulty in this regard has been the introduction of cyclosporine therapy in the treatment of autoimmune diseases and other diseases affecting the skin. For example, in the treatment of atopic dermatitis and psoriasis, and as broadly suggested in the art, as a hair growth stimulating use, for example, in the treatment of alopecia caused by old age or disease.

Thus, oral cyclosporine therapy shows that the drug has significant potential benefits, for example, for patients suffering from psoriasis, while at the risk of side effects from oral therapy, preventing universal use. Various proposals have been made in the art for the application of cyclosporin, for example cyclosporin in topical formulations and many topical delivery systems have been described. However, several attempts at topical application have failed to provide noticeable effective therapies.

However, the present invention overcomes the problems described above herein. More specifically, the prodrug of the present invention significantly improves its solubility in aqueous solutions when compared to a pharmaceutical non-prodrug formulation. When administered in solution, there is no need to use a carrier such as ethanol or castor oil. In addition, the prodrugs of these agents do not exhibit the side effects of the formulations in the prior art, according to the present invention. Furthermore, it has now been found that many of the agents in the table below are administered in their prodrug form by the present invention, thereby improving oral absorption, thereby significantly improving its bioavailability and efficacy.

Preferred agents used in combination with amino acids form the prodrugs listed here below in the following table, and the advantages found are listed in the second column at the end of the table. In the table, the key points are:

a) improved taste, smell

b) desired octanol / water partition coefficient (ie, solubility in water / fat)

c) improved stability in vitro and in vivo

d) enhanced penetration of the blood-brain barrier

e) elimination of first-pass effects in the liver

f) reduction of intestinal recirculation

g) painless injections in parenteral formulations

h) improved bioavailability

i) increase in absorption

j) reduced side effects

k) proportionality of dosage

1) Selective Hydrolysis of Prodrugs at the Site of Action

m) controlled release properties

n) targeted drug delivery

o) reduced toxicity, thereby improving therapeutic rates

p) reduced dosage

q) alteration of metabolic pathways to deliver more agents to the site of action

In addition, the table indicates the usefulness of prodrugs. The utility of the prodrug is the same as the corresponding medicament (with no amino acid moiety bound). The usefulness is described in literature such as [Physicians Desk Reference, 2004 edition], the contents of which are incorporated by reference.

5.

The following non-limiting examples further illustrate the present invention:

Synthesis of Various Amino Acid Derivatives of Selected Agents

I. Propofol Derivatives

Propofol (2,6-diisopropylphenol is a low-molecular-weight phenol that is widely used as a central nervous system anesthetic and has a sedative and hypnotic effect. The drug is used intravenously to induce anesthesia and / or to maintain anesthesia in mammals. The main advantages of propofol are that the drug is quickly induced to anesthesia, has minimal side effects, and when the drug is discontinued, the patient quickly recovers without prolonged sedation.

Propofol has been shown to be quite fluctuating and somewhat surprising in many therapeutic applications, for example the drug has been shown to be an effective antioxidant, anti-vomiting agent, anti-pruritic agent, anti-epileptic drug, anti-inflammatory drug. And even appear to have anticancer properties.

Mechanism of action:

The mechanism of action of propofol has been extensively studied, and its central nervous system anesthesia has been shown to be related to the high affinity of the drug for a particular subclass of GABA receptors (Collins GGS, 1988, Br. J. Pharmacology. 542,225). -232). However, there are numerous other types of receptors in the brain that are substrates for propofol, whereby the activity of the agent changes.

Propofol also has important biological effects as an antioxidant. Because of this generalized action of propofol, the drug is theoretically useful in the treatment of numerous inflammatory processes in which oxidation is an important factor. For example, there is the synthesis of cyclooxygenase mediated prostaglandin, which causes inflammation. Propofol in the treatment of acid aspiration, adult / infant respiratory distress syndrome, airway obstructive diseases, asthma, cancer and many other similar pathological conditions by inhibiting oxidation in the respiratory pathway Can be used.

It is very common for tissue injury to occur due to oxidation, so propofol can cause Parkinson's disease, Alzheimer's disease, Fredrich's disease, Huntington's disease, and multiple sclerosis. It has been claimed that it may be useful for the treatment of sclerosis, amyotrophic lateral sclerosis, spinal cord injuries, and various other neurodegenerative diseases.

Propofol is currently marketed by Astra Zenaca under the brand name Diprivant in the United States. It is one of the most widely used short-acting central nervous system anesthetics currently on the market. The concentration of propofol is 10 mg / mL in pyrogen-free sterile emulsions and the formulation contains soybean oil, glycerol, egg lecithin, disodium edetate and sodium hydroxide.

An important disadvantage of propofol is the fact that the agent is completely insoluble in water. Even at very low concentrations of 10 mg / mL, the agent settles out of the aqueous solution at room temperature. Therefore, the producers of these formulations use bold methods to emulsify these products in water with very unusual complexes and toxic emulsifiers. For example, producers of intravenous formulations of the medicament use egg lecithin, Cremephore L®, castor oil, and other similar emulsifiers.

However, the use of such emulsifiers is associated with many problems. It is due to the fact that various types of cremapoel emulsifiers can promote allergic reactions. Egg lecithin and castor oil have been shown to cause anaphylactic shock in some patients. In addition, maintaining the stability of propofol in these emulsions leads to shorter and more expensive. In addition, the presence of egg lecithin and castor oil is made to facilitate the growth of fresh water in the emulsion. It would probably be possible to dissolve propofol in water by combining propofol with cyclodextrin, but cyclodextrin has not been approved by the FDA for use in intravenous therapy.

So far, no one has made a safe propofol prodrug. British Patents 1,102,011 and 1,160,468, and US Pat. No. 3,389,138 describe various phenolic esters of amino acids, where propofol is bound to many side chains that produce toxic effects when released into the human body.

US Pat. No. 6,451,854 describes many substituted alpha amino acetate esters of propofol, where propofol and the side chain were substituted with many different kinds of chemical groups. All N, N-disubstituted glycine esters of propofol have been shown to be non-toxic and many of the compounds described therein are derivatives of propofol. Thus, when esters are released into the body after cleavage by enzymes, many of the active agents are not propofol, whereby they do not have data indicating that they are toxic and have no known therapeutic efficacy in humans as a whole. Have new molecules.

Another published article on the water-soluble salts of the amino acid esters of propofol, an anesthetic, [Int. In J. Pharmaceutics, 175 [2]: 195-204, 1998, the authors synthesized many water soluble derivatives of propofol. However, when these prodrugs are cleaved by esterase enzymes, substituted unnatural amino acids with unknown toxicity profiles are released into the human body.

To date, no pharmaceutical formulation is available that can deliver propofol without any deleterious side effects. However, the present invention has produced many propofol derivatives that are water soluble and non-toxic, which make propofol without any harmful side effects and the need for toxic and expensive additives, solulizers and emulsifiers. Suitable for delivery into the human body without

Thus, in one aspect, the present invention relates to a class of prodrugs of propofol. The prodrug consists of a carboxyl group of amino acids esterified with free hydroxyl groups present on propofol molecules.

More specifically, one aspect of the present invention relates to compounds having the following formulae or pharmaceutically acceptable salts thereof,

Where AA is an amino acid and the carboxyl group of AA reacts with the carboxyl group of propofol.

In another aspect, the invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of the various propofol prodrugs above and their pharmaceutical carriers.

In another embodiment, the invention is directed to a method of treating a patient in need of propofol therapy, the method comprising administering to the patient an effective amount of propofol. .

In another embodiment, the present invention is directed to a method of improving the solubility of propofol in an aqueous solution comprising reacting with hydroxyl functional groups of propofol to separate their products.

In another embodiment, the present invention, when administered to a patient, selects to reduce or eliminate potential toxic side effects of current formulations containing toxic excipients and to form respective ester covalent bonds of the propofol molecule. The present invention relates to a substantially and therapeutically effective method comprising reacting a carboxyl group with a hydroxyl group of amino acids and separating their products to administer the product to the patient.

The present invention provides that when naturally occurring unsubstituted amino acids are esterified to propofol, the resulting prodrugs are very water soluble (> 200 mg / L in water), break down in the human body and release non-toxic amino acids and are toxic. Shows that none of the emulsifiers, additives and other excipients are required.

Furthermore, the present invention also produced drugs, which are the prodrugs of the propofol of the present invention and are very effective central nervous system anesthetics. Thus, amino acid prodrugs of the present invention are effective CNS anesthetics that release or do not release an effective parent drug.

The amino acid esters of the present invention are at least 10 times more soluble than propofol dissolved in water at room temperature. In particular, the glycine, proline and lysine esters of propofol are soluble in the range higher than 100 mg / ml and higher than 250 mg / mL for lysine.

The prodrugs of the present invention are not expected to have any antioxidant action due to the interference of phenolic groups leading to the following phenomena; However, the inventors have discovered that prodrugs of propofol are effective anesthetics with or without propofol. The prodrugs of propofol described above release propofol when administered in an in vivo experiment and the resulting agent retains its pharmacological and antioxidant properties.

The prodrugs of propofol of the present invention clearly offer many advantages over propofol, for example, all of the cleaved side chains in these prodrugs are naturally occurring essential amino acids and are nontoxic by the fact.

These facts lead to high therapeutic indices. Secondly, all prodrugs are easily broken into the body to release propofol. Furthermore, because of the high water solubility of the prodrugs, they can be easily administered, forming solutions in - situ , or infusion, just prior to intravenous injection with lyophilized sterile powder. To the prefilled syringes or bottles. Amino acid esters are more stable than propofol because the OH groups in propofol interfere with oxidation. Thus, the propofol prodrugs of the present invention are more effective than propofol itself, which is non-toxic and free of other pharmaceutical problems associated with currently available formulations.

Prodrugs of propofol of the present invention have anti-inflammatory, anti-oxidative, anti-cancer, anti-convulsive, anti-emetic, and anti-pruritic properties.

The prodrugs of propofol of the present invention are effective in the treatment of diseases or disorders in which propofol is normally used. The prodrugs disclosed herein enhance the therapeutic advantages of propofol by reducing or eliminating the biopharmaceutical and pharmacokinetic barriers that are modified in the human body to release the active compound and are associated with each of the prodrugs. However, it should be noted that these prodrugs themselves will have sufficient activity in mammals without releasing any active agent. Since the prodrugs are more soluble in water than propofol, they do not need to be associated with a carrier vehicle, such as alcohol or castor oil, which may be toxic or cause unwanted additional reactions. In addition, oral formulations containing the prodrugs of propofol are absorbed into the blood and are quite effective.

Thus, the prodrugs of the present invention enhance the therapeutic advantages by removing the biopharmaceutical and pharmacokinetic barriers of existing agents.

Furthermore, the prodrugs are easily synthesized in high yields easily and using currently available materials.

summary:

Glycine, L-proline of propofol. And the procedure for the synthesis of esters of L-lysine is shown here below. However, these esters are illustrative examples and their amino acid prodrugs can be prepared using the following methods. Complete process and analytical data are provided in the "experimental part". In general, as shown in the following scheme, propofol (10 g) has a catalytic capacity of 4- (N, N-dimethylamino) -pyridine (4- (N, N-dimethyamino) -pyridine) (DMAP). In presence, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide), combined with N-Boc protecting amino acid (1 equivalent) with hydrochloride (EDC) It became. EDC was extracted with water and removed. After drying with sodium sulphate, filtration and concentration, the amino acid esters of unprocessed and protected propofol were purified by flash chromatography to yield protected esters in 50-60% yield. The protecting groups were then removed by stirring the protected esters in diethyl ether liquid saturated with gaseous hydrochloride at room temperature. The yield of the deprotection step was generally 60-95%. After filtration and drying, the hydrochloride salts of glycine and L-proline esters of propofol did not require further purification. The hydrochloride salt of the L-lysine ester of propofol was crystallized from ethanol once to remove traces of the mono-protected L-lysine-propofol ester.

Synthesis order :

plan

Glycine, L-proline of propofol. And synthesis of esters of L-lysine: a) EDC, DMAP, CH ₂ C1 ₂ ; b) HC1 (g), Et ₂ 0.

Part of the experiment:

The synthesis of SPI0010, SPI0011 and SPI0013 was performed in one batch. In general, small scale experiments were performed initially and then as larger batches. Reagents mentioned in the experimental section were purchased with the highest purity available from Sigma, Aldrich, Acros or Bachem, except for solvents purchased from either Fisher Scientific or Mallinkrodt.

One) SPI0010

Propofol (9.98 g, 55.97 mmole) was dissolved in dichloromethane (200 mL) at room temperature under argon atmosphere. Nt-Butyloxocarbonyl-glycine (Nt-Butyloxocarbonyl-glycine) (11.2 g, 63.91 mmole) is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, hydrochloride (EDC, 11. lg, 57.9 mmole) and 4- (N, N-dimethylamino) -pyridine (DMAP, 1.5 g, 12.27 mmole). At room temperature, under argon atmosphere, after stirring for 21 hours, water (200 mL) was added and the layers separated. The dichloromethane layer was washed again with water (200 mL) and dried over sodium sulfate (5 g) for 1 hour. After filtration and concentration under reduced pressure, the remaining oil was purified by flash chromatography on silica gel (250 g), eluting with hexane / ethyl acetate (10: 1). . The process produced a glycine ester of protected N-BOC protected propofol as a white solid (11.34 g, 60% yield).

(tert-butoxycarbonylamino-acetic acid 2,6-diisoprosylphenyl ester):

¹ H NMR (300 MHz, CDC1 _{3 )} : δ = 7.25-7.13 (m, 3H), 5.18 (br s, 1H), 4.22 (d, 2H, J = 5.7 Hz), 2.89 (m, 2H), 1.46 (s, 9H), 1.18 (d, 12H, J = 6.9 Hz).

¹³ C NMR (75 MHz, CD1 ₃ ): δ = 169.35, 155.75, 145.22, 140.35, 126.90, 124.14, 80.32, 42.66, 28.54, 27.79, 23.57.

Propofol-Boc-gycine ester (11.28 g, 33.6 mmole) was dissolved in anhydrous diethyl ether (200 mL) at room temperature. Hydrochloride (gas) was penetrated into the solution and stirring continued for 45 minutes. The mixture was stirred for 48 hours at room temperature under argon atmosphere. After 48 hours hexanes (200 mL) were added and the precipitate was filtered off. The white solid was dried under high pressure vacuum at 88 ° C. for 5 hours. The experiment produced SPI0010 (8.73 g, 95% yield, using 99.9% purity HPLC) as a white solid.

Amino-acetic acid 2,6-diisopropyl-phenyl ester, hydrochloride:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 8.77 (br s, 3H), 7.20-7.08 (m, 3H), 4.14 (m, 2H), 2.87 (m, 2H), 1.11 (d, 12H, J = 7 Hz).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 166.42, 144.84, 140.42, 127.10, 124.06, 40.47, 27.61, 23.55.

CHN analysis: calc. : C 61.87, H 8.16, N 5.15; found: C 61.14, H 8.20, N 5.14.

2) SPI0011

Propofol (10.03 g, 56.23 mmole) was dissolved in dichloromethane (100 mL) at room temperature under argon atmosphere. Nt-butyloxocarbonyl-glycine (14.04 g, 65.22 mmole) is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, hydrochloride (EDC, 11.95 g, 62.33 mmole) and 4- (N, With N-dimethylamino) -pyridine (DMAP, 1.1 g, 9.0 mmole). At room temperature, under argon atmosphere, after stirring for 3 hours, water (100 mL) was added and the layers separated. The dichloromethane layer was washed again with water (100 mL) and dried over sodium sulfate (5 g) for 1 hour. After filtration and concentration under reduced pressure, the remaining oil was purified by flash chromatography on silica gel (250 g), eluting with hexane / ethyl acetate (10: 1). The process produced the proline ester of protected N-BOC protected propofol as stand up in the refrigerator to yield a solidified clear oil (11.34 g, 66% yield).

Pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester 2- (2,6-diisopropyl-phenyl) ester;

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.31-7.20 (m, 3H) 4.73 (m, 1H), 3.70-3.50 (m, 2H), 3.20-2.94 (m, 2H), 2.46-2.20 ( m, 2H), 2.20-2.0 (m, 2H), 1.55 (m, 9H), 1.25 (m, 12H).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 171.87, 171.01, 154.34, 153.93, 145.35, 145.23, 140.06, 140.21, 126.69, 126.53, 123.95, 80.28, 79.89, 59.14, 46.67, 46.42, 31.10, 30.17, 28.61 , 28.56, 28.56, 27.44, 27.18, 23.47.

Propofol-Boc-glycine ester (13.95 g, 37.14 mmole) was dissolved in anhydrous diethyl ether (100 mL) at room temperature. Hydrochloride (gas) was penetrated into the solution and stirring continued for 60 minutes. The mixture was stirred for 22 hours at room temperature under argon atmosphere. After 22 hours hexanes (50 mL) were added and the precipitate was filtered off. The white solid was dried under high pressure vacuum at 88 ° C. for 5 hours. The experiment produced SPI0011 (9.1 g, 81% yield, using 99.1% purity HPLC) as a white solid.

Pyrrolidine-2 (S) -carboxylic acid 2,6-diisopropyl-phenyl ester, hydrochloride:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 10.15 (br s, 2H), 7.27-7.14 (m, 3H), 4.78 (t, 1H, J = 7.8 Hz), 3.56 (m, 2H), 2.85 (m, 2H), 2.64 (m, 1H), 2.40 (m, 1H), 2.20 (m, 1H), 2.05 (m, 1H), 1.18 (m, 12H).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 168.30, 144.23, 139.74, 126.98, 123.96, 51.58, 38.21, 29.32, 26.64, 26.18, 23.71, 23.02, 21.67

CHN analysis: calc. : C 65.48, H 8.40, N 4.49; found: C 65.50, H 8.43, N 4.50.

3) SPI0013

Dicyclohexylamine salt (23.62 g, 0.0447 mole) of di-N-boc-L-lysine is diethyl ether (200 ml) and potassium hydrogen sulfate (potassium). hydrogen sulfate) (9.14 g) was added in chilled water (200 ml) in an ice / water bath. After stirring for 20 minutes, the layers separated. The ether layer was extracted three times with cold water (100 ml). The ether layer was dried over sodium sulfate (15 g) for 1 hour, filtered and concentrated under reduced pressure. The process produced free acid (15.5 g, 100% recovery) of N, N-di-boc-L-lysine.

Propofol (8.0 g, 45 mmole) was dissolved in dichloromethane (100 mL) at room temperature under argon atmosphere. N, N'-di-t-butyloxocarbonyl-glycine (15.5 g, 44.7 mmole) is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, hydrochloride (EDC, 8.62 g, 45 mmole) ) And 4- (N, N-dimethylamino) -pyridine (DMAP, 0.55 g, 4.5 mmole). At room temperature, under argon atmosphere, after stirring for 3 hours, water (100 mL) was added and the layers separated. The dichloromethane layer was washed again with water (100 mL) and dried over sodium sulfate (5 g) for 1 hour. After filtration and concentration under reduced pressure, the remaining oil was purified by flash chromatography on silica gel (250 g), eluting with hexane / ethyl acetate (9: 1). The process produced a proline ester of protected N-BOC protected propofol in a white foam (12.42 g, 55% yield).

2 (S), 6-Bis-t-butoxycarbonylamino-hexanoic acid 2,6-diisopropyl-phenyl ester:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.28-7.15 (m, 3H), 5.22 (d, 1H, J = 8.4 Hz), 4.70 (m, 1H), 4.59 (m, 1H), 3.17 ( m, 2H), 2.93 (m, 2H), 2.09 (m, 1H), 1.86 (m, 1H), 1.67-1.54 (m, 4H), 1.48 (s, 9H), 1.46 (s, 9H), 1.20 (m, 12 H).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 171.82, 156.10, 155.65, 145.25, 140.30, 126.80, 124.03, 80.14, 79.28, 53.76, 40.29, 32.09, 28.66, 28.54, 27.48, 23.91, 23.10.

Propofol-di-Boc-L-lycine ester (12.34 g, 24.37 mmole) was dissolved in anhydrous diethyl ether (250 mL) at room temperature. Hydrochloride (gas) was penetrated into the solution and stirring continued for 60 minutes. The mixture was stirred for 48 hours at room temperature under argon atmosphere. After 48 hours the precipitate was filtered off and crystallized from ethanol (100 ml). The white solid was dried under high pressure vacuum at 90 ° C. for 4 hours. The experiment produced SPI0013 (5.5 g, 60% yield, using 98.6% purity HPLC) as a white solid.

2 (S), 6-Diamino-hexanoic acid 2,6-diisopropyl-phenyl ester, dihydrochloride:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 9.05 (br s, 3H), 8.35 (br s, 3H), 7.26-7.13 (m, 3H), 4.43 (t, 1H, J = 6 Hz), 3.0-2.6 (m, 4H), 2.09 (m, 2H), 1.80-1.50 (m, 4H), 1.10 (d, 12H, J = 7 Hz).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 168.30, 144.23, 139.74, 126.98, 123.96, 51.58, 38.21, 29.32, 26.64, 23.71, 23.02, 21.67.

CHN analysis: calc. : C 56.99, H 8.50, N 7.38; found: C 56.48, H 8.56, N 7.30.

II . Nonsteroidal  Prodrugs of anti-inflammatory drugs NSAIDs )

Non-steroidal anti-inflammatory agents include structurally distinct classes of carboxylic acid moieties attached to planar aromatic functional groups, examples include: acetyl salicylic acid, salicylic acid, diflunisal, ibuprofen, pefe Nopropene, carpropene, flurbiprofen, ketoprofen, naproxen, sulindac, indomethacin, indodolac, tolmetin Ketorolac, diclofenac and meclofenamate. Nonsteroidal anti-inflammatory drugs have anti-inflammatory, analgesic, antipyretic and anti-clotting action.

Examples of the chemical structure of this unique class of compounds with broad pharmacological activity are shown below.

Nonsteroidal anti-inflammatory drugs are widely used as anticoagulant drugs for the treatment of acute and chronic pain, the management of tissue injury and edema from inflammatory joint diseases, and also for the treatment of myocardial infarction. Many of them are analgesic and anti-inflammatory in addition to antipyretic action, which is useful for reducing fever.

Several drugs in this group have also been prescribed for Rheumatoid Arthritis, Osteoarthritis, acute gout, ankylosing spondylitis, and dysmenorrhea.

Mechanism of action:

 It is a major mechanism for nonsteroidal anti-inflammatory drugs to produce their therapeutic efficacy through inhibition of prostaglandin synthesis. Specifically, nonsteroidal anti-inflammatory drugs inhibit cyclooxygenases such as COX-1 and COX-2 enzymes, in which case these two enzymes are responsible for the synthesis of prostaglandins. While COX-1 enzymes are important for the regulation of platelet aggregation, blood flow in the kidneys, and gastric acid secretion, COX-2 enzymes play an important role in pain and inflammatory processes. Nonsteroidal anti-inflammatory drugs can be used to prevent thromboembolism and myocardial infarction by significantly increasing clotting time.

All nonsteroidal anti-inflammatory drugs are relatively moderate to strong organic acids with pKa levels in the 3-6 range. Most of them are carboxylic acid derivatives. Acidic groups are essential in terms of COX inhibitory activity and physiological pH. All nonsteroidal anti-inflammatory drugs ionize. They all have a fairly variable hydrophilic / hydrophobic balance, and these agents have the functionality of their aryl groups, aromatic side chains and aliphatic side chains, and other heterocyclic variants in their structures.

Most nonsteroidal anti-inflammatory drugs bind so much with plasma proteins and often replace other drugs with competitive affinities for plasma proteins. Thereby, co-administration of nonsteroidal anti-inflammatory drugs with other therapeutic class of drugs must be carefully evaluated to prevent drug interactions. Most of the drug is metabolized in mammals through conjugation because of its acidic carboxyl group. The major route of metabolic clearance of many nonsteroidal anti-inflammatory drugs is glucuronidation following renal elimination.

The use of acetylsalicylic acid (aspirin) in coronary heart diseases is now well known and it has been proven to be a lifesaver for many patients with myocardial infarction. . Several additional uses have already been made in the literature on aspirin, for example, the fact that aspirin reduces the risk of stroke in patients with early warning symptoms of a potential ischemic heart attack. Reported in the journal Lancet (Vol 349, p 1641). Pre-eclampsia and fetal growth retardation are both due to the occlusion of blood vessels in the placenta, two of the most common complications of pregnancy-there are millions of cases of preeclampsia worldwide each year. . In an experiment involving more than 9,000 women in 16 countries, 60 mg of aspirin daily reduced the risk of preeclampsia by 13 percent. Aspirin has also been shown to be effective in several studies to prevent colon cancer, lung cancer and pancreatic cancer in postmenopausal women. Because aspirin can improve blood flow, its usefulness is becoming increasingly evident in the treatment of certain forms of dementia, such as diabetes and Alzheimer's disease.

Because of its unique pharmacological potential, nonsteroidal anti-inflammatory drugs have attracted considerable attention in the media. The primary area of clinical investigation of these drugs has been as nonsteroidal anti-inflammatory drugs, especially in patients suffering from pain, arthritis, (rheumatic and bone joints), other inflammatory reactions, and fever. The application of drugs and the prevention of coronary heart diseases. These drugs are also used to treat migraines, menstrual syndromes, back pain and gout.

Despite the very major contributions made by nonsteroidal anti-inflammatory drugs, the development of unwanted side effects reported has been reported-especially in severe gastric ulcer and duodenal ulcers, mucosal erythema, and edema, erosion, perforation, bloody stool, ulcer Colitis has obviously been a serious obstacle to their widespread use or application-as well as more effective and convenient methods of administration (eg galenic formulations, eg providing adequate bioavailability and providing appropriate and controlled Not only are the dosages available at an accelerated rate, but are also convenient and provide difficulty in providing oral dosage forms for patients). The dual injury theory includes NSAID mediated direct injury following the systemic effects of prostaglandin synthesis being inhibited. Local injury also occurs as a result of bile secretion of active liver metabolites and subsequent reflux of the duodenum stomach (Arthritis and Rheumatism 1995; 38 (1): 5-18); the effects are additional; Either local or systemic mechanism alone is sufficient to cause mucosal injury of the gastrointestinal duodenum.

In addition, the aforementioned nonsteroidal anti-inflammatory drugs are very hydrophobic and easily settle even where very small amounts of water are present. E.g. This is true even if it comes into contact with the human body (for example, gastric juice). Thus, for example. In terms of form and taste, it is very difficult to provide oral formulations that are easy to administer to a patient, are stable when stored, and that can be administered as usual habits to provide appropriate and patient controlled dosages.

The oral administration of the presented liquid formulations, eg, nonsteroidal anti-inflammatory drugs, is to date based in principle on the use of natural rubbers such as Xanthan, cellulose, citric acid, and lime flavors. See, for example, US Pat. No. 5,780, 046. Drink-solutions of non-steroidal anti-inflammatory drugs currently on the market are incompatible with orange pigments and strawberry flavors, citric acid, xanthan gum, polysorbate 80, gelatin pregelatinized starch, glycerin, sodium benzoate (sodium benzoate), and added artificial colors and flavors. The use of the drink solution and similar compositions as presented in this technique involves a number of difficulties.

Furthermore, the good taste of existing oil base systems has proved to be a problem. The taste of the existing drink-solutions is in particular unpleasant. Mixing into a moderately flavored drink is done, for example, in a chocolate drink formulation, usually at high dilution prior to dosing, to enable normal treatment to an acceptable level. The use of oil-based systems also requires the use of high ethanol concentrations, which are themselves inherently undesirable, particularly where pediatric administration is expected. In addition, evaporation of ethanol is for example. From capsules (used in many parts, to solve the problem of good taste, as discussed or as other formulations (eg when opened)), the development of nonsteroidal anti-inflammatory drug sedimentation is made. When such a composition is presented, for example, in a soft gelatin encapsulation formulation, this particular difficulty is in the packaging of an air-tight component, for example an airtight transparent pack or aluminum foil blister pack, an encapsulated product. need. This in turn makes the goods bulkier, producing more expensive production. The storage characteristics of the formulations mentioned above are, in addition, still far from ideal.

Gastrointestinal irritation of nonsteroidal anti-inflammatory drugs is a subject of great concern not only to patients but also to physicians. Acute use of aspirin, phenopropene, plubiprofen, indomethacin, ketodolac, meclofenamate, mefenamic acid, and pyroxicam cause serious gastrointestinal side effects. Even ibuprofen has been shown to cause severe gastrointestinal lesions with prolonged use. Gastrointestinal toxicity is the most frequently encountered side effect associated with nonsteroidal anti-inflammatory drugs and is of considerable concern. About half of all hospitalized patients with hemorrhagic ulcers are due to nonsteroidal anti-inflammatory drugs, aspirin, or a combination of both during the week prior to hospitalization [Faulkner G, Prichard P, Somerville K, et al. Aspirin and bleeding peptic ulcers in the elderly. Br Med J. 1988; 297: 1311-1313. Investigations by patients in Tennessee Medicaid who were hospitalized for gastrointestinal complications showed that patients using nonsteroidal anti-inflammatory drugs had gastrointestinal bleeding or peptic ulcer disease than patients who did not take nonsteroidal anti-inflammatory drugs. The risk was about four times greater [Griffin MR, Piper JM, Daugherty JR, et al. Nonsteroidal anti-inflammatory drug use and increased risk for peptic ulcer disease inelderly persons. Ann Intern Med. 1991; 114: 257-263. Severe gastrointestinal outbreaks occur 2% to 4% per year in patients undergoing continuous nonsteroidal anti-inflammatory medications for rheumatoid arthritis, according to the FDA. Gastrointestinal ulcers (4.725), duodenal ulcers (1.1 to 1.6), bleeding (3.8), perforation, and relative risk of death all use nonsteroidal anti-inflammatory drugs when compared to humans whose patients do not take these products Is increased by. In 1989, patients with rheumatoid arthritis were admitted approximately 20,000 times per year and an estimated cost of $ 10,000 per hospitalization was used [Fries JF, Miller SR, Spitz PW, et al. Toward an epidemiology of gastropathy associated with nonsteroidal anti-inflammatory drug use. J Gastroenterology. 1989; 96: 647-655.

There is also a need to provide some nonsteroidal anti-inflammatory drugs in water-soluble formulations for use as injections. It is well known that alcohols and tromethamine used to form salts in current formulations of ketodolak are toxic. At present there is no formulation which allows nonsteroidal anti-inflammatory drugs to be in an aqueous solution at those concentrations required due to the low water solubility of the drug.

Above all such very obvious practical difficulties, there is the occurrence of unwanted additional reactions already mentioned, which are observed using the applicable oral dosage forms.

Several visible suggestions for solving these various problems have been presented in this technology, including oral dosage forms of both solid and liquid. However, the primary difficulty still remaining here is that the nonsteroidal anti-inflammatory drugs shown in the table below are insoluble in aqueous media inherently, due to their nature, dosage forms that may contain non-steroidal anti-inflammatory drugs in sufficiently high concentrations. The use of is avoided. The formulation is convenient to use but yet still meets the required criteria in terms of bioavailability, for example, to achieve effective absorption from the gastrointestinal or intestinal lumen consistent with adequately high blood / serum concentrations. It is a formulation.

The prodrugs of the nonsteroidal anti-inflammatory drugs of the present invention overcome the problems mentioned above herein. More specifically, one embodiment of the present invention relates to a prodrug of a nonsteroidal anti-inflammatory drug, which significantly improves its solubility in aqueous solutions and therefore, when administered in solution, There is no need to use. In addition, the prodrugs of nonsteroidal anti-inflammatory drugs do not exhibit the side effects that have been preceded by this technology by the present invention. Furthermore, the prodrugs of the present invention almost completely avoid the gastrointestinal irritation seen upon oral administration and therefore significantly improve the therapeutic index and their efficacy of the tested prodrugs.

Thus, in one aspect, the invention relates to prodrugs of nonsteroidal anti-inflammatory drugs. Predrugs or pharmaceutically acceptable salts of the preferred nonsteroidal anti-inflammatory drugs have the formula

In the formula, Y is NH-AA or O-AA and AA is an amino acid, wherein either the amine group or hydroxyl group of AA reacts with the carboxylic acid group of the nonsteroidal anti-inflammatory drugs.

The present invention also relates to pharmaceutical compositions comprising a therapeutically effective amount of the various nonsteroidal anti-inflammatory agents and pharmaceutical carriers thereof described above.

In another embodiment, the present invention is directed to a method of treating a patient in need of nonsteroidal anti-inflammatory drug therapy, the method comprising administering to the patient described above an effective amount of non-steroidal anti-inflammatory drugs.

In another embodiment, the present invention is directed to a method of improving the solubility of nonsteroidal anti-inflammatory drugs in an aqueous solution, comprising reacting the carboxyl functional groups of individual nonsteroidal anti-inflammatory drugs to separate their products. do.

In another embodiment, the present invention is directed to a method in a practical and therapeutically effective manner, wherein when administered to a patient to reduce or eliminate injury to the gastrointestinal mucosa of nonsteroidal anti-inflammatory drugs, the individual The nonsteroidal anti-inflammatory drug molecule is reacted with one of two of the amine or hydroxyl functional groups of the selected amino acids to form one of two of the amide or ester covalent bonds, respectively, and to separate their products to administer the above product to the patient. It involves doing.

A. Synthesis of Ibuprofen Amino Acid Derivatives

summary:

The procedure for the synthesis of L-serine, L-threonine, and L-hydroxyproline esters of ibuprofen will be outlined in the Synthesis Sequence section. The complete process and analytical data will be provided in the experimental section. Also, these plans are illustrative examples. This scheme is applicable to other amino acids in the preparation of prodrugs of the nonsteroidal anti-inflammatory drugs of the invention. In general, (±)-ibuprofen (4-10 g, in batch) is 1- (3-dimethyl in the presence of a catalytic capacity of 4- (N, N-dimethylamino) -pyridine (DMAP) Aminopropyl) -3-ethylcarbodiimide, hydrochloride (EDC, 1 equiv) combined with N-benzyloxybenzyl ester protecting amino acids (1 equiv). After the reactions were completed, some excess EDC was extracted and removed by water, DMAP was removed by extraction with diluted acid, and ibuprofen was removed by extraction with sodium bicarbonate. After drying over sodium sulfate, filtration and concentration, the amino acid esters of (±) -ibuprofen, which have not yet been processed and protected, are used immediately or otherwise flash chromatography to produce protected esters in good yields (85-95%). Purified by chromatography. If a slight excess of ibuprofen and a coupling agent were used, column chromatography would not normally be necessary, and the entire extraction process was performed. The protecting groups were removed by hydrogenation (25-35 psi hydrogen) in the presence of 10% palladium in carbon and hydrochloric acid. The yield of the deprotection step was generally in the range of 70-90%. After filtration and drying, the hydrochloride salts of serine and threonine esters of (±) -ibuprofen were purified by crystallization reaction. No further purification was required. The hydrochloride salt of the L-hydroxyproline ester of ibuprofen was in the form of a gel that could be solidified or crystallized. In this case the hydrogenation reaction was repeated without the use of acid and the neutral compound was purified.

Since ibuprofen began with a mixture of enantiomers, the final product was delivered as a mixture of stereoisomers except for threonine esters. In the case of the threonine ester of ibuprofen, washing with water, acetone, or acetonitrile could easily separate the final stereoisomeric salts. Insoluble isomers (SPI0016A) were determined to be valid isomers by comparison with certified standards made from S (+)-ibuprofen. Serine and hydroxyproline esters of (±) -ibuprofen could not be easily separated during this process.

Synthetic sequence:

One. SPI0015

2. SPI0016A Wow SPI0016B

3. SPI0017

Synthesis of L-serine, L-threonine, and L-hydroxyproline esters of (±) -ibuprofen: a) EDC, DMAP, CH ₂ Cl ₂ ; b) HCl, 10% Pd / C, EtOH c) Acetone, d) 10% Pd / C, EtOH.

Part of the experiment:

Synthesis of SPI0015, SPI0016 and SPI0017 was performed in two or three batches. The materials mentioned in the experimental section were purchased with the highest purity that can be obtained from Sigma-Aldrich, Acros or Bachem, except for solvents purchased from either Fisher Scientific or Mallinkrodt.

1) Preparation of (±) -Ibuprofen-L-serine ester, hydrochloride (SPI0015).

(±) -Ibuprofen (5.04 g, 24.4 mmole), N-carbenzyloxy-L-serine benzyl ester (8.11 g, 24.6 mmole), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydro Chloride (EDC, 4.87 g, 25.4 mmole), and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.40 g, 3.27 mmole) were dissolved in dichloromethane (150 mL) at room temperature under argon atmosphere . At room temperature, under argon atmosphere, after stirring for 22 hours, water (100 mL) was added and the layers separated. The dichloromethane layer was washed again with water (100 mL) and dried over sodium sulfate (5 g) for 1 hour. After filtration and concentration under reduced pressure, the remaining oil was purified by flash chromatography on silica gel (250 g), eluting with hexane / ethyl acetate (3: 1). The procedure produced protected L-serine- (±) ibuprofen ester (SPI001501) as a colorless solid (11.4 g, 90% yield).

2 (S) -Benzyloxycarbonylamino-3- [2 (R, S)-(4-isobutyl-phenyl) -propionyloxyl-propionic acid benzyl ester:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.40-7.20 (m, 10H), 7.14-7.01 (m, 4H), 5.50 (d,% 1 / 2H, J = 8.4 Hz), 5.29 (d, 1 / 2H, J = 8.4 Hz), 5.11-5.02 (m, 2.5H), 4.90 (d, l / 2H, J = 12 Hz), 4.62 (m, 1H), 4.49-4.43 (m, 1H), 4.36-4.32 (m, 1H), 3.59 (m, 1H), 2.39-2.35 (m, 2H), 1.78 (m, 1H), 1.42-1.39 (m, 3H), 0.85 (d, 6H, J = 6.6 Hz).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 174.05, 169.19, 169.07, 155.68, 140.73, 137.20, 136.12, 135.05, 134.91, 129.44, 128.67, 128.65, 128.60, 128.41, 128.33, 128.30, 128.19, 127 1619 , 67.75, 67.32, 64.51, 64.32, 53.71, 45.16, 45.02, 30.35, 22.60, 18.27.

Protected ibuprofen-L-serine ester (22.50 g, 43.4 mmole) was dissolved in ethanol (200 mL) at room temperature, and under nitrogen atmosphere, pars containing 10% palladium (3.86 g, 50% wet) on carbon. Poured into a Parr bottle. Hydrochloric acid (30 mL of water to 10 mL of 37% HCl) was added and replaced with hydrogen gas (25 psi) under nitrogen atmosphere. After stirring for 4 hours, the palladium catalyst was removed by filtration through celite. The ethanol / water was removed under reduced pressure. The remaining white solids were washed with water (25 mL), acetone (20 mL) and then dried under high pressure vacuum conditions (4 h at 88 ° C.). This experiment produced (±) -ibuprofen-L-serine ester, hydrochloride (SPI0015) (11.3 g, 80% yield) as a colorless solid.

2 (S) -Amino-3- [2 (R, S)-(4-isobutylphenyl) -propionyloxy] -propionic acid, hydrochloride; ((R, S)-ibuprofen-L-serine ester, hydro Chloride):

¹ H NMR (300 MHz, DMSO): δ = 8.92 (br s, 3 H), 7.22 (t, 2H, J = 7.5 Hz), 7.10 (d, 2H, J = 7.5 Hz), 4.56 (m, 1H) , 4.37-4.20 (m, 2H), 3.83 (q, 1H, J = 6.9 Hz), 2.41 (d, 2H, J = 6.9 Hz), 1.80 (m, 1H), 1.41 (d, 3H, J = 6.9 Hz), 0.85 (d, 6H, J = 6.9 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 173.36, 173.32, 168.08, 168.04, 139.70, 128.96, 129.92, 127.20, 127.05, 62.47, 51.59, 51.49, 44.28, 44.00, 43.90, 29.68, 22.28, 18.70, 18.42

HPLC analysis: 99.13% purity; rt = 3.133 min; Luna C18 5u column (sn 167917-13); 4.6 x 250 mm; 254 nm; 50% ACN / 50% TFA buffer (0.1%); 35 C; 20 ul inj .; lml / min; 1 mg / mL sample size; Sample dissolved in mobile phase.

CHN analysis: calc. : C 58.27, H 7.33, N 4.25; found: C 58.44, H 7.46, N 4.25.

Melting Point: 169.5-170.5 ℃

2a) ( ± ) -Ibuprofen -L-threonine ester, hydrochloride ( SPI0016A And SPI0016B) preparation and separation method.

(±)-ibuprofen (4.15 g, 20.11 mmole), N-carbenzyloxy-L-threonine benzyl ester (6.90 g, 20.11 mmole), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, Hydrochloride (EDC, 3.95 g, 20.6 mmole), and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.25 g, 2.0 mmole) are dissolved in dichloromethane (50 mL) at room temperature under argon atmosphere It became. After stirring for 19 hours at room temperature, under argon atmosphere, the dichloromethane layer was again washed with water (50 mL), 5% hydrochloric acid (2x25 mL), water (25 mL), saturated sodium bicarbonate (2x25 mL), and water (50 mL). After drying for 1 hour over sodium sulfate (5 g), filtration and concentration under reduced pressure, the remaining oil was used as is without further purification. The process produced a protected L-threonine- (±)-ibuprofen ester (SPI001601) (10.2 g, 95.3% yield) as a pale yellow fat that solidified on standing.

2 (S) -benzyloxycarbonylamino-3- [2 (R, S)-(4-isobutyl-phenyl) -propionyloxy] -butyl acid benzyl ester:

¹ H NMR (300 MHz, CDCl ₃ ): = 7.40-7.15 (m, 10H), 7.14-7.01 (m, 4H), 5.48-5.25 (m, 2H), 5.11-5.01 (m, 3H), 4.90 ( d, 1 / 2H, J = 12 Hz), 4.68 (d, 1 / 2H, J = 12 Hz), 4.48 (m, 1H), 3.60-3.48 (m, 1H), 2.39 (m, 2H), 1.79 (m, 1H), 1.42-1.35 (m, 3H), 1.27 (d, 1.5H, J = 6.6 Hz), 1.17 (d, 1.5H, J = 6.6 Hz), 0.85 (m, 6H).

¹³ C NMR (75 MHz, CDCl ₃ ): δ = 173.32, 169.70, 169.30, 156.55, 140.75, 137.38, 137.22, 136.14, 135.07, 134.99, 129.45, 129.41, 128.65, 128.39, 128.22, 127.21, 127.14, 70.70, 70. , 67.81, 67.66, 67.53, 57.83, 45.19, 30.39, 22.61, 18.57, 18.30, 17.18, 16.87.

Protected ibuprofen-L-threonine ester (10.15 g, 19.0 mmole) was dissolved in warm ethanol (150 mL) at room temperature and contained 10% palladium (3.4 g, 50% wet) in carbon under nitrogen atmosphere. Pour into the bottle. Hydrochloric acid (6 mL 37% HCl in 20 mL water) was added and the nitrogen atmosphere was replaced with hydrogen gas (30 psi). After stirring for 3 hours, the palladium catalyst was removed by filtration through celite (30 g). The ethanol / water was removed under reduced pressure. This experiment produced (±) -ibuprofen-L-threonine ester, hydrochloride (SPI0016A and SPI0016B, 6.4 g, 97% crude yield) as a colorless solid.The original mixture of stereoisomers was acetone (200 mL). The solids (2.84 g, SPI0016A) were filtered off After 2 hours, the filtered product (SPI0016B, 3.0 g) was concentrated under reduced pressure.

1.) Purification of SPI0016A (active isomer):

After three batches of S-Ibuprofen-L-threonine ester (SPI0016A) were completed, the batches were mixed (8.78 g total) and then crystallized three times from DIUF water (100 mL). Each time, a small amount of zwitterion was produced. To regenerate the salts, the resulting solids (in each crystallization reaction) were dissolved in 1% hydrochloric acid solution in ethanol (3 mL 37% hydrochloric acid in 100 mL ethanol). The ethanol solution was then concentrated at room temperature under reduced pressure. After the third crystallization process and regeneration process, the salt (5.6 g) was stirred in acetonitrile (100 mL) for 44 hours at room temperature, under argon atmosphere. The salt was then filtered and then dried under high pressure vacuum conditions at 50-55 ° C. until its weight was constant (5.5 g).

2 (S) -Amino-3 (R)-[2 (S)-(4-isobutyl-phenyl) -propionyloxy] -butyl acid; (S-Ibuprofen-L-threonine ester, hydrochloride, active isomer):

¹ H NMR (300 MHz, DMSO): δ = 8.76 (br s, 3H), 7.19 (d, 2H, J = 8.1 Hz), 7.11 (d, 2H, J = 8.1 Hz), 5.28 (dq, 1H, J = 6.3, 3.6 Hz), 4.14 (q, 1H, J = 3.6 Hz), 3.80 (q, 1H, J = 7.2 Hz), 2.41 (d, 2H, J = 7.2 Hz), 1.80 (m, 1H) , 1.37 (d, 3H, J = 7.2 Hz), 1.21 (d, 3H, J = 6.3 Hz), 0.85 (d, 6H, J = 6.6 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 172.66, 168.24, 139.68, 137.24, 128.95, 126.97, 67.98, 55.35, 44.23, 43.83, 29.66, 22.24, 18.52, 16.47.

CHN analysis: calc .: C 59.38, H 7.62, N 4.07; found: C 59.17, H 7.63, N 4.04.

HPLC analysis: 98.28% purity; rt = 6.951 min .; 60% TFA (0.1%) / 40% acetonitrile; 1 mL / min; 37.5 C; Luna C18, 3u column (SN 167917-13), 4.6 × 250 mm; 22 ul injection.

Optical rotation: + 24.5 °

Melting point: 189-190 ℃

2) Purification of SPI0016B (Inactive Isomers):

After three batches of R-ibuprofen-L-threonine ester (SPI0016B) were completed, the batches were mixed (9.02 g total) and then crystallized from DIUF water (50 mL). During the crystallization reaction, a small amount of zwitterion was produced. To regenerate the salts, the resulting solids (in each crystallization reaction) were dissolved in 1% hydrochloric acid solution in ethanol (3 mL 37% hydrochloric acid in 100 mL ethanol). The ethanol solution was then concentrated at room temperature under reduced pressure. After the third crystallization reaction and regeneration process, the salt (5.93 g) was crystallized three times by adding a small amount in acetone (1 mL) from hot toluene (100 mL). The salt was then filtered and then dried under high pressure vacuum conditions at room temperature until its weight was constant (5.1 g).

2 (S) -Amino-3 (R)-[2 (R)-(4-isobutyl-phenyl) -propionyloxy] -butyl acid; (R-Ibuprofen-L-threonine ester, hydrochloride, inert isomers):

¹ H NMR (300 MHz, DMSO): δ = 8.82 (br s, 3H), 7.23 (d, 2H, J = 7.8 Hz), 7.10 (d, 2H, J = 7.8 Hz), 5.27 (m, 1H) , 4.18 (m, 1H), 3.80 (q, 1H, J = 7.2 Hz), 2.41 (d, 2H, J = 7.2 Hz), 1.81 (m, 1H), 1.41 (d, 3H, J = 6.9 Hz) , 1.34 (d, 3H, J = 6.3 Hz), 0.85 (d, 6H, J = 6.3 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 72.56, 168.08, 139.64, 136.98, 128.84, 127.14, 68.8, 55.29, 44.28, 29.69, 22.28, 18.24, 16.41.

CHN analysis: calc .: C 59.38, H 7.62, N 4.07; found: C 59.30, H 7.60, N 4.05.

HPLC analysis: 98.43% purity; rt = 6.19 min .; 60% TFA (0.1%) / 40% acetonitrile; 1 mL / min; 37.5 C; Luna C18, 3u column (SN 167917-13), 4.6 × 250 mm; 22 ul injection.

Optical rotation: + 10.4 °

Melting Point: 176-177 ° C

2b) S-(+)-Ibuprofen-L-threonine ester, hydrochloride Standard Preparation (SPI0016S).

S-(+)-Ibuprofen (2.0 g, 9.69 mmole), N-carbenzyloxy-L-threonine benzyl ester (3.25 g, 9.91 mmole), 1- (3-dimethylaminopropyl) -3-ethylcarbodii Meade, hydrochloride (EDC, 1.90 g, 9.91 mmole), and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.12 g, 1.0 mmole) were dichloromethane (25 mL) at room temperature under argon atmosphere. Dissolved in. After stirring for 4 hours, the dichloromethane layer was washed again with water (25 mL), 5% hydrochloric acid (25 mL), saturated sodium bicarbonate (2x25 mL), and water (25 mL). After drying for 1 hour over sodium sulfate (5 g), filtration and concentration under reduced pressure, the remaining oil was used as is without further purification. The process yielded a protected S-(+)-ibuprofen-L-threonine ester (SPI001601S) (5.01 g, 98% yield) as a preferentially solidified pale yellow oil.

2 (S) -benzyloxycarbonylamino-3- [2 (R, S)-(4-isobutyl-phenyl) -propionyloxy] -butyl acid benzyl ester:

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.35-7.23 (m, lOH), 7.10 (d, 2H, J = 7.8 Hz), 7.05 (d, 2H, J = 7.8 Hz), 5.48-5.25 ( m, 2H), 5.17-5.01 (m, 4H), 4.50 (dd, 1H, J = 9.6, 1.8 Hz), 3.50 (q, 1H, J = 7.2 Hz), 2.40 (d, 2H, J = 7.2 Hz ), 1.80 (m, 1H), 1.37 (d, 3H, J = 7.2 Hz), 1.17 (d, 3H, J = 6.3 Hz), 0.86 (d, 6H, J = 6.6 Hz).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 173.29, 169.69, 156.51, 140.68, 137.21, 136.08, 135.06, 129.40, 128.70, 128.66, 128.57, 128.38, 128.24, 127.14, 70.70, 67.80, 67.53, 57.87, 45.19 , 45.11, 30.39, 22.61, 18.57, 16.87.

Protected S-(+)-Ibuprofen-L-threonine ester (5.0 g, 9.40 mmole) was dissolved in warm ethanol (100 mL) at room temperature, and under nitrogen atmosphere, 10% palladium (1.0 g, 50) on carbon. poured into a Parr bottle containing% wet). Hydrochloric acid (1 mL of 37% HC1 in 10 mL of water) was added and the nitrogen atmosphere was replaced with hydrogen gas (32 psi). After stirring for 2 hours, the palladium catalyst was removed by filtration through celite (30 g). The ethanol / water was removed under reduced pressure. This experiment produced (±) -ibuprofen-L-threonine ester, hydrochloride (SPI0016S, 2.8 g, 85% raw yield) as a colorless solid. The original mixture of stereoisomers was stirred in acetone (50 mL) for 3 hours at room temperature under argon atmosphere. After 3 hours the solids (2.24 g, 69% yield) were filtered and then dried under high pressure vacuum conditions at room temperature until the weight was constant.

2 (S) -Amino-3 (R- [2 (S)-(4-isobutyl-phenyl) -propionyloxy] -butyl acid;

(S-Ibuprofen-L-threonine ester, hydrochloride, active isomer):

¹ H NMR (300 MHz, DMSO): δ = 8.76 (br s, 3H), 7.19 (d, 2H, J = 8.1 Hz), 7.11 (d, 2H, J = 8.1 Hz), 5.28 (dq, 1H, J = 6.3, 3.6 Hz), 4.14 (q, 1H, J = 3.6 Hz), 3.80 (q, 1H, J = 7.2 Hz), 2.41 (d, 2H, J = 7.2 Hz), 1.80 (m, 1H) , 1.37 (d, 3H, J = 7.2 Hz), 1.21 (d, 3H, J = 6.3 Hz), 0.85 (d, 6H, J = 6.6 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 172.66, 168.24, 139.68, 137.24, 128.95, 126.97, 67.98, 55.35, 44.23, 43.83, 29.66, 22.24, 18.52, 16.47.

HPLC analysis:

98.28% purity; rt = 6.951 m; 60% TFA (0.1%) / 40% acetonitrile; 1 ml / min; 37.5 C Luna C18, 3u column (SN 1679017-13), 4.6 × 250 mm; 22 ul injection

Optical rotation: + 26.5 °

Melting point: 189-190 ℃

 3) (±) -Ibuprofen-L- Hydroxyproline  Preparation of ester ( SPI0017 ).

(±) -Ibuprofen (5.10 g, 24.7 mmole), N-carbobenzyloxy-L-hydroxyproline benzyl ester (8.80 g, 24.7 mmole), 1- (3-dimethylaminopropyl) -3-ethylcarbodii Meade, hydrochloride (EDC, 5.10 g, 26.0 mmole), and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.30 g, 2.40 mmole) were dichloromethane (100 mL) at room temperature under argon. Dissolved in. At room temperature, under argon atmosphere, after stirring for 24 hours, water (100 mL) was added and the layers separated. The dichloromethane layer was washed again with water (100 mL) and 5% sodium bicarbonate (2x50 mL). After drying for 1 hour over sodium sulfate (5 g), filtration and concentration under reduced pressure, the remaining oil was used as is without further purification. The process produced a protected (±)-ibuprofen-L-hydroxyproline ester (SPI001701) (11.5 g, 85% yield) as pale yellow oil.

4 (R)-[2- (4-Isobutyl-phenyl) -propionyloxy] -pyrrolidine-2 (S) -carboxylic acid;

((R, S) -Ibuprofen-L-hydroxyproline ester):

¹ H NMR (300 MHz, CDC13): δ = 7.33-7.02 (m, 14H), 5.25-4.95 (m, 5H), 4.51-4.19 (m, 1H), 3.75-3.50 (m, 3H), 2.40 ( d, 2H, J = 6.9 Hz), 2.15 (m, 1H), 1.81 (m, 1H), 1.44 (d, 3H, J = 7.0 Hz), 0.87 (d, 6H, J = 6.6 Hz).

¹³ C NMR (75 MHz, CDCl ₃ ): S = 173.99, 171.93, 171.72, 154.68, 154.15, 140.70, 137.23, 137.04, 136.23, 135.44, 135.23, 129.41, 128.59, 128.47, 128.35, 128.19, 128.08, 127.89, 127.02 , 72.86, 72.16, 67.40, 67.18, 67.09, 58.12, 57.83, 52.66, 52.49, 52.13, 45.15, 36.63, 35.67, 32.07, 30.33, 29.23, 22.90, 22.58, 18.36.

Protected ibuprofen-L-hydroxyproline ester (11.40 g, 43.4 mmole) was dissolved in warm ethanol (150 mL) at room temperature and, under nitrogen atmosphere, 10% palladium (2.73 g, 50% wet) in carbon. It was poured into a par jar containing it. The nitrogen atmosphere was replaced with hydrogen gas (34 psi). After stirring for 5 hours, the palladium catalyst was removed by filtration through celite. The ethanol was removed under reduced pressure. The remaining white solids (6.60 g) were washed with DIUF water (50 mL), diethyl ether (50 mL) and then dried under high pressure vacuum conditions until the weight was constant. This experiment produced (±) -ibuprofen-L-hydroxyproline ester SPI0017 (5.64 g, 84% yield) as a colorless solid.

4 (R)-[2- (4-Isobutyl-phenyl) -propionyloxy] -pyrrolidine-2 (S) -carboxylic acid;

((R, S) -Ibuprofen-L-hydroxyproline ester):

¹ H NMR (300 MHz, CDC13): 6 = 7.22 (d, 2H, J = 7.2 Hz), 7.09 (d, 2H, J = 7.2 Hz), 5.27 (m, 1H), 4.40 (t, 0.5H, J = 7 Hz), 4.24 (t, 0.5 H, J = 9 Hz), 3.75 (m, 1H), 3.61 (m, 1H), 3.28 (d, 0.5H, J = 13 Hz), 3.15 (d, 0.5H, J = 13 Hz), 2.42-2.10 (m, 4H), 1.78 (m, 1H), 1.40 (br t, 3H, J = 6 Hz), 0.82 (d, 6H, J = 6 Hz). (Mixture of diastereomers)

¹³ C NMR (75 MHz, CDC13): δ = 173.28, 173.23, 168.98, 139.88, 137.33, 137.23, 129.12, 127.26, 127.17, 72.58, 57.60, 57.50, 50.24, 50.12, 44.34, 44.15, 29.31, 34.16, 34.16 22.34, 18.43, 18.23. (Mixture of isomers)

HPLC analysis: 100% purity; rt = 5.35, 5.22 min. ; 55% TFA (0.1%), 45% ACN; 1 mL / min; 32.3 C, Luna C18, serial # 188255-37; 20 ul inj ..

CHN analysis: calc .: C 67.69, H 7.89, N 4.39; found: C 67.47, H 7.87, N 4.30.

Melting Point: 198-199 ℃

Efficacy of the synthesis of (±) -Ibuprofen's L-serine, L-threonine, and L-hydroxyproline esters by using acetylcholine induced abdominal contraction in male albino mice (anti-invasive water potential):

The present study contemplates the antagonistic properties of acetylcholine-induced writhing as an index in Albino mice, thereby L-hydrin of L-serine, L-threonine, and (±) -ibuprofen. It was performed to evaluate the efficacy of the oxyproline esters. Ibuprofen (racemic mixture) and ibuprofen (S)-(+) served as reference controls.

Other new formulations of ibuprofen and the reference control, ie ibuprofen (racemic mixture) and ibuprofen (S)-(+), were 5% (v / v) in milli Q water as the vehicle. Using Tween 80, male albino rats (Swiss strain) were administered via feeding gavage. This study was performed at two doses, 50 mg and 100 mg / kg body weight, with vehicle control. Ten animals were used at each dose. All doses were expressed in molar equivalents of ibuprofen. The molar equivalents as well as the dosages used are given below.

Formulation: Molar equivalent Formulation Molar equivalent S-(+)-Ibuprofen-L-threonine ester 0.833 units is equivalent to 1 unit of ibuprofen. (±) -Ibuprofen-L-serine ester 1.6 units is equivalent to 1 unit of ibuprofen. (±) -Ibuprofen-L-hydroxyproline ester 1.55 units is equivalent to 1 unit of ibuprofen.

Test Item: Group: Dose (mg / kg): Equivalent Weight of Test Item Test Items group Dose (mg per kg) [in terms of ibuprofen] Equivalent Weight of Test Items [mg / kg] Vehicle Vehicle control 0.0 - S-(+)-Ibuprofen-L-threonine ester Trial group 1 50.0 41.65 Test group 2 100.0 83.30 (±) -Ibuprofen-L-serine ester
(Ibuprofen S) Test group 3 50.0 80.0 Test group 4 100.0 160.0 (±) -Ibuprofen-L-hydroxyproline ester Trial group 5 50.0 77.5 Test group 6 100.0 155.0 Ibuprofen (racemic mixture) Test group 7 50.0 50.0 Test group 8 100.0 100.0 Ibuprofen S + Test group 9 50.0 25.0 Test group 10 100.0 50.0

Efficacy in terms of antagonistic effects on one acetylcholine-induced writhing at three doses and two doses of 50.0 and 100.0 mg / kg for the reference control are presented below.

Test Item: Group: Dose (mg / kg): Number of animals showing no single struggle Test Items
group
Dose (mg per kg) [in terms of ibuprofen]
Number of animals showing no single struggle (number of animals per dose = 10) 1 hour after dosing 3 hours after dosing Vehicle Vehicle control 0.0 0 0 S-(+)-Ibuprofen-L-threonine ester
Low dose 50.0 One 0 High dose 100.0 3 0 (±) -Ibuprofen-L-serine ester
Low dose 50.0 4 2 High dose 100.0 6 4 (±) -Ibuprofen-L-hydroxypropolin ether Low dose 50.0 5 4 High dose 100.0 7 7 Ibuprofen (racemic mixture)
Low dose 50.0 4 2 High dose 100.0 6 6 Ibuprofen S + Low dose 50.0 5 One High dose 100.0 6 6

Statistical analysis using the Chi-square test procedure did not show any statistically significant differences between the new formulations when compared to the reference control, whereas the comparison with the number of animals There was no writhing in the groups, and the relative "p" was found to be greater than 0.05, which is a significant level.

From clinical observations based on that number of animals showing no writhing by administration of acetylcholine, (±) -Ibuprofen-L-hydroxyproline esters were formulated in different formulations and ibuprofen (racemic) and ibuprofen (S)-( It has been found to be more effective in antagonizing acetylcholine-induced writhing compared to +).

Summary of efficacy of L-serine, L-threonine, and L-hydroxyproline esters of (±) -ibuprofen, ibuprofen (racemic mixture) and ibuprofen (S)-(+) based on antagonism of acetylcholine induced writhing in albino rats Dose (mg per kg) [in terms of ibuprofen] Test Items
Number of animals showing no single struggle (number of animals per dose = 10) 1 hour after dosing 3 hours after dosing



50 mg / kg Vehicle control 0 0 S-(+)-Ibuprofen-L-threonine ester One 0 (±) -Ibuprofen-L-serine ester 4 2 (±) -Ibuprofen-L-hydroxypropolin ether 5 4 Ibuprofen (racemic mixture) 4 2 Ibuprofen (S)-(+) 5 One

100 mg / Kg Vehicle control 0 0 S-(+)-Ibuprofen-L-threonine ester 3 0 (±) -Ibuprofen-L-serine ester 6 4 (±) -Ibuprofen-L-hydroxypropolin ether 7 7 Ibuprofen (racemic mixture) 6 6 Ibuprofen (S)-(+) 6 6

Data was presented by statistical analysis using the chi-square test to assess the efficacy of the new formulations compared to the reference control. The experiment showed no statistically significant difference between the new formulations when compared to the reference control, whereas the comparison to the number of animals showed no writhing in each group, and the relative "p" Was found to be greater than 0.05, which can be said to be a significant level.

The data is also summarized in FIGS. 1 and 2. (±) -Ibuprofen-L-hydroxyproline ester from a bar diagram of efficacy that is comparable to clinical observations based on a number of animals that do not exhibit struggles due to the administration of acetylcholine (FIGS. 1 and 2) Has been found to be more effective in antagonizing acetylcholine-induced writhing compared to other formulations and ibuprofen (racemic) and ibuprofen (S)-(+).

conclusion

The study of the present invention was conducted to assess the relative efficacy of the new formulation of ibuprofen. For this reason, this antagonistic nature of new formulations in acetylcholine induced struggles was chosen as an index to determine the relative efficacy of the formulations. Ibuprofen (racemic mixture) and ibuprofen (S)-(+) served as reference controls. This study was performed at two doses, 50 and 100 mg / kg, with vehicle control.

Efficacy is shown below in terms of antagonistic effects on one acetylcholine-induced writhing at three doses and two doses 50.0 and 100.0 mg / kg for the reference control.

Test Item: Group: Dose (mg / kg): Number of animals showing absence of single struggle (out of 10) Test Items
group
Dose (mg per kg) [in terms of ibuprofen]
Number of animals showing no single struggle (number of animals per dose = 10) 1 hour after dosing 3 hours after dosing Vehicle Vehicle control 0.0 0 0 S-(+)-Ibuprofen-L-threonine ester
Low dose 50.0 One 0 High dose 100.0 3 0 (±) -Ibuprofen-L-serine ester
Low dose 50.0 4 2 High dose 100.0 6 4 (±) -Ibuprofen-L-hydroxypropolin ether Low dose 50.0 5 4 High dose 100.0 7 7 Ibuprofen (racemic mixture)
Low dose 50.0 4 2 High dose 100.0 6 6 Ibuprofen (S)-(+) Low dose 50.0 5 One High dose 100.0 6 6

Statistical analysis using the Chi-square test procedure did not show any statistically significant differences between the new formulations when compared to the reference control, whereas the comparison with the number of animals There was no writhing in the groups, and the relative "p" was found to be greater than 0.05, which is a significant level.

However, clinical observations based on that number of animals that do not show writhing by the administration of acetylcholine show that (±) -ibuprofen-L-hydroxyproline ester has different formulations and ibuprofen (racemic) and ibuprofen (S). It has been found to be more effective in antagonizing acetylcholine-induced writhing compared to-(+).

(±) - gastric mucosal irritation potential of the ibuprofen in the L- serine, L- threonine, and L- hydroxyproline ester fasting state of male albino rats of

summary

This study is responsible for gastrointestinal mucosal irritation / lesion in fasting male albino rats in new formulations of ibuprofen ((±) -Ibuprofen L-serine, L-threonine, and L-hydroxyproline esters). This was done to determine its relative potential. Ibuprofen (racemic mixture) and ibuprofen (S)-(+) served as reference controls.

New formulations of ibuprofen and other types of reference controls, namely ibuprofen (racemic mixture) and ibuprofen (S)-(+), were used in male Albino rats (Swiss) using 5% Tween 80 as a carrier in milli-Q water. strain was administered via a feeding gavage. The study was performed at two doses, with the vehicle control at 200 mg and 300 mg / kg body weight. Five animals were used at each dose. All doses were expressed in molar equivalents of ibuprofen. The molar equivalents as well as the dosages used are given below.

Formulation: Molar equivalent Formulation Molar equivalent S-(+)-Ibuprofen-L-threonine ester 0.833 units is equivalent to 1 unit of ibuprofen. (±) -Ibuprofen-L-serine ester 1.60 units is equivalent to 1 unit of ibuprofen. (±) -Ibuprofen-L-hydroxyproline ester 1.55 units is equivalent to 1 unit of ibuprofen.

The various groups used are labeled here:

Test Item: Group: Dose (mg / kg) Equivalent Weight Test Items group Dose (mg per kg) [in terms of ibuprofen] Equivalent Weight of Test Items [mg / kg] Vehicle Vehicle control 0.0 - S-(+)-Ibuprofen-L-threonine ester Trial group 1 200.0 0.0 Test group 2 300.0 166.6 (±) -Ibuprofen-L-serine ester
(Ibuprofen S) Trial group 1 200.0 249.9 Test group 2 300.0 320.0 (±) -Ibuprofen-L-hydroxyproline ether Trial group 1 200.0 480.0 Test group 2 300.0 310.0 Ibuprofen (racemic mixture) Trial group 1 200.0 465.0 Test group 2 300.0 300.0 Ibuprofen (S)-(+) Trial group 1 200.0 100.0 Test group 2 300.0 150.0

Mice are fasted on a cycle of 18 to 22 hours prior to administration. The test item was administered as a single dose via nutrient gavage. For three hours after drug administration, the animals were allowed to die humanly through inhalation of carbon dioxide gas. The stomach was removed and the following were observed.

 ● amount of mucosal exudate,

 ● redness and thickening of the stomach wall

 The location of the bleeding (local or sporadic), the nature of the bleeding according to its size (petechial or orecchymotic)

 ● perforation or any other disorder

The observations of gastrointestinal mucosa irritation in various groups of animals are summarized below.

Test Item: Group: Observation of Dose (mg / kg) Test Items group Dose mg per kg (for ibuprofen) observe Vehicle control Vehicle control 0.0 None of the animals showed any evidence of gastric mucosal irritation S-(+)-Ibuprofen-L-threonine ester Trial group 1 200.0 None of the animals showed any evidence of gastric mucosal irritation Test group 2 300.0 None of the animals showed any evidence of gastric mucosal irritation (±) -Ibuprofen-L-serine ester Trial group 1 200.0 None of the animals showed any evidence of gastric mucosal irritation Test group 2 300.0 None of the animals showed any evidence of gastric mucosal irritation (±) -Ibuprofen-L-hydroxyproline ether Trial group 1 200.0 None of the animals showed any evidence of gastric mucosal irritation Test group 2 300.0 None of the animals showed any evidence of gastric mucosal irritation Ibuprofen (racemic mixture) Trial group 1 200.0 Gastric mucosal irritation was observed in one of the five animals administered Test group 2 300.0 Gastric mucosal irritation was observed in two of the five animals administered. Ibuprofen (S)-(+) Trial group 1 200.0 Gastric mucosal irritation was observed in all five animals administered. Test group 2 300.0 Gastric mucosal irritation was observed in three of the five animals administered.

The results of this study showed that no formulations of ibuprofen were responsible for any evidence of irritation to the gastrointestinal mucosa in male fasting albino rats at the two doses tested (200 mg and 300 mg / kg body weight). . In contrast, both ibuprofen (racemic mixture) and ibuprofen (S)-(+) both caused irritation to the gastrointestinal mucosa at both doses tested. Furthermore, ibuprofen (S)-(+) was found to be more irritating to the gastrointestinal mucosa than ibuprofen (racemic mixture).

Ketoprofen  S (+) threonine ester synthesis overview:

The procedure for the synthesis of L-threonine esters of ketoprofen will be outlined in the Synthesis Sequence section. These schemes are illustrative examples and can be applied equally to other amino acids. The complete process and analytical data will be provided in the experimental section. In general, (±) -ketoprofen (5 g) is N-boc-L threonine t-butyl ester ¹ (in the presence of a catalytic capacity of 4- (N, N-dimethylamino) -pyridine (DMAP). 1 equivalent) and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide, hydrochloride (EDC). After the reactions were complete, any excess EDC was extracted and removed with water, DMAP was removed by extraction with diluted acid, and ketoprofen was removed by extraction with sodium bicarbonate. After drying over sodium sulfate, filtration and concentration, the raw, yet protected L-threonine- (±) -ketoprofen flash flash to produce protected L-threonine esters in good yield (98%). Purified to The protecting groups were removed with 2M hydrochloric acid in diethyl ether to split the boc group and then treated with trifluoroacetic acid to remove t-butyl ester. After the drying process, a mixture of L-threonine-R, S (±) -ketopropene esters was separated from the acetonitrile by crystallization reaction. Hydrochloride of L-threonine-S (+) ketopropene ester preferentially precipitates from acetonitrile. Visually pure samples were prepared for comparison and started with S (+)-ketoprofen. After drying and analysis, L-threonine-S (+) ketopropene ester, hydrochloride (1.75 g) was separated from the mixture.

Synthesis order :

SPI0018A

Synthesis of L-threonine esters of ketoprofen : a) EDC, DMAP, CH ₂ Cl ₂ ; b) HCl (2M); c) TFA; d) ACN (crystallization).

Part of the experiment:

Synthesis of SPI0018A was performed in a single batch. The agents mentioned in the experimental section were purchased with the highest purity available from Sigma, Aldrich, Acros or Bachem, except for solvents purchased from either Fisher Scientific or Mallinkrodt.

S (+) - ketoprofen -L- threonine ester, the method and isolation of the hydrochloride (SPI0018A).

(±) -ketoprofen (5.32 g, 20.92 mmol), Nt-butylcarbonyl-L-threonine t-butyl ester (Boc-Thr-OtBu, 5.17 g, 18.72 mmol, (made according to literature), 1 -(3-dimethylaminopropyl) -3-ethylcarbodiimide, hydrochloride (EDC, 4.0 g, 20.9 mmol), and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.22 g) at room temperature , Under argon atmosphere, was dissolved in dichloromethane (50 mL) After stirring for 5 h, the dichloromethane layer was again water (50 mL), 5% hydrochloric acid (2x25 mL), water (25 mL), saturated bicarbonate Washed with sodium (2x25 mL), and water (50 mL), dried over sodium sulfate (5 g) for 1 h, filtered and concentrated under reduced pressure, the remaining oil (10.3 g) was silica gel. (150 g), purified by column chromatography eluting with hexanes / ethyl acetate (2: 1) The product containing fractions were mixed, concentrated and After the high pressure after drying under vacuum conditions, the process is a protected L- threonine were produced for the ketoprofen ester as a clear oil (SPI001801) (9.42 g, 98% yield) - (±).

3- [2 (R, S)-(3-Benzoyl-phenyl) -propionyloxy] -2 (S) -tert-butoxycarbonylamino-butyl acid tert-butyl ester: (mixture of diastereomers)

¹ H NMR (300 MHz, CDC13): δ = 7.83-7.42 (m, 9H), 5.43 (dd, 1H, J = 13.2, 6.9 Hz), 5.10 (dd, 1H, J = 20.7, 9.3), 4.29 ( t, 1H, J = 11.7 Hz), 3.75 (q, 1H, J = 7.2 Hz), 1.50-1.42 (m, 19.5H), 1.30-1.18 (m, 4.5H).

¹³ C NMR (75 MHz, CDC13): δ = 196.18, 172.62, 172.55, 168.85, 168.58, 155.81, 140.33, 140.23, 137.86, 137.39, 132.46, 132.42, 131.54, 131.38, 130.00, 129.354, 129.13, 129.02 128.27, 82.50, 82.37, 80.05, 71.38, 71.22, 57.59, 57.52, 45.46, 45.31, 28.40, 27.98, 27.84, 18.54, 18.48, 17.19, 16.84.

Protected (R, S) -ketopropene-L-threonine ester (9.42 g, 18.41 mmol) was dissolved in dichloromethane (25 mL) at room temperature under argon atmosphere. Anhydrous hydrochloric acid (2M, 25 mL) in diethyl ether was added to the solution, and the mixture was stirred at room temperature for 17 hours. The mixture was concentrated under reduced pressure. The remaining foam (8.2 g) was dissolved in a mixture of dichloromethane (10 mL) and trifluoro acetic acid (20 mL). At room temperature, after stirring for 6.5 hours the solution was concentrated under reduced pressure. Toluene (25 mL) was added to the remaining oil and the mixture was concentrated a second time. Anhydrous hydrochloric acid (2M, 20 mL) in ethanol (20 mL) and diethyl ether was added to the solution, and the mixture was concentrated third. After drying under high pressure vacuum conditions at room temperature for 2 hours, this experiment was carried out yellowish (±) -ketoprofen-L-threonine ester, hydrochloride (mixture of stereoisomers, 7.11 g, 98% raw yield). It was produced as an off-white solid. A crude mixture of stereoisomers (7.0 g) was crystallized three times with acetonitrile (200 mL). After the third crystallization reaction, the remaining white solid was dried under high vacuum at 50 ° C. until its weight became constant (4 hours). This experiment produced L-threonine-S (+)-ketopropene ester, hydrochloride SPI0018A (2.2 g, 30% yield from SPI001801).

2 (S) -Amino-3 (R)-[2 (S)-(3-benzoyl-phenyl) -propionyloxy] -butyl acid, hydrochloride (L-threonine-S (+)-ketopropene ester Hydrochloride):

¹ H NMR (300 MHz, DMSO): δ = 14.08 (br s, 1H), 8.72 (br s, 3 H), 7.74-7.51 (m, 9H), 5.29 (t, 1H, J = 4.5 Hz), 4.16 (m, 1H), 3.97 (q, 1H, J = 6.3 Hz), 1.42 (d, 3H, J = 6.9 Hz), 1.23 (d, 3H, J = 6.3 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 195.34, 172.26, 168.21, 140.42, 137.05, 136.74, 132.66, 131.66, 129.48, 128.73, 128.49, 128.30, 68.23, 55.31, 44.00, 18.44, 16.45.

CHN analysis: calc .: C 61.30, H 5.66, N 3.57; found: C 61.02, H 5.58, N 3.58.

HPLC analysis: 98.28% purity; rt = 25.14 min. ; 55% DIUF water (0.1% TFA) / 45% Methanol; 1 mL / min; 36.4 C; Luna C18, 5u column (serial # 211739-42), 4.6 × 250 mm; 20 ul injection.

Optical rotation: + 27.0 ° (20 C, 174.4 mg / 10 mL ethanol, 589 nm); Melting point: 166-167 ℃

S-(+)- ketoprofen- L-threonine ester, hydrochloride The law of the Standard.

(+)-Ketoprofen (1.87 g, 7.74 mmol), Nt-butylcarbonyl-L-threonine t-butyl ester (Boc-Thr-OtBu, 2.25 g, 8.14 mmol, prepared by literature method), 1 -(3-dimethylaminopropyl) -3-ethylcarbodiimide, hydrochloride (EDC, 1.65 g, 8.60 mmol), and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.1 g) at room temperature Dissolved in dichloromethane (25 mL) under argon atmosphere. After stirring for 4 hours, the dichloromethane layer was washed again with water (25 mL). After drying for 1 hour over sodium sulfate (5 g), filtration and concentration under reduced pressure, the remaining oil was used as is without further purification. The process produced a protected L-threonine-(+)-ketopropene ester (4.01 g, -100% yield) as pale yellow oil.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.81-7.42 (m, 9H), 5.43 (m, 1H), 5.10 (d, 1H, J = 9.3), 4.29 (d, 1H, J = 9.6 Hz ), 3.75 (q, 1H, J = 7.2 Hz), 1.50-1.42 (m, 21H), 1.18 (d, 3H, J = 6.3 Hz).

¹³ C NMR (75 MHz, CDCl ₃ ): δ = 196.4, 172.79, 168.99, 155.94, 140.44, 137.99, 137.51, 132.59, 131.50, 130.13, 129.31, 129.25, 129.15, 128.66, 128.40, 82.68, 80.24, 71.37, 57.71, 57.71 , 45.43, 28.53, 28.10, 18.99, 16.96.

Protected (S) -ketopropene-L-threonine ester (3.92 g, 7.66 mmol) was dissolved in anhydrous hydrochloric acid (2M, 50 mL) in diethyl ether and then stirred at room temperature for 17 hours. The mixture was concentrated under reduced pressure. The remaining foam (3.4 g) was dissolved in a mixture of dichloromethane (20 mL) and trifluoro acetic acid (20 mL). At room temperature, after stirring for 6.5 hours the solution was concentrated under reduced pressure. Toluene (25 mL) was added to the remaining oil and the mixture was concentrated a second time. Anhydrous hydrochloric acid (2M, 20 mL) in ethanol (20 mL) and diethyl ether was added to the solution, and the mixture was concentrated third. After drying under high pressure vacuum conditions at room temperature for 2 hours, the experiment showed that (+)-ketoprofen-L-threonine ester, hydrochloride (3.05 g crude) was a yellowish white solid (off) -white solid). The raw materials were stirred with acetone (50 mL) at room temperature for 2 hours. The remaining white solid was filtered and then dried under high vacuum at 50 ° C. until its weight was constant (4 hours). This experiment produced L-threonine-S (+)-ketopropene ester, hydrochloride (2.04 g, 67% yield).

¹ H NMR (300 MHz, DMSO): b = 14.08 (br s, 1 H), 8.72 (br s, 3 H), 7.74-7.51 (m, 9 H), 5.29 (t, 1 H, J = 4.5 Hz), 4.16 (m, 1H), 3.97 (q, 1H, J = 6.3 Hz), 1.42 (d, 3H, J = 6.9 Hz), 1.23 (d, 3H, J = 6.3 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 195.34, 172.26, 168.21, 140.42, 137.05, 136. 74, 132.66, 131.66, 129.48, 128.73, 128.49, 128.30, 68.23, 55.31, 44.00, 18.44, 16.45.

HPLC analysis: 99.43% purity; rt = 25.14 min. ; 55% DIUF water (0.1% TFA) / 45% methanol; 1 mL / min; 36.4 C; Luna Cl 8, 5u column (serial # 211739-42), 4.6 × 250 mm; 20 ul injection.

Optical rotation: + 27.1 ° (20 ° C., 177.8 mg / 10 mL ethanol, 589 nm); Melting point: 166-167 ℃

 C. Amino Acid Derivatives of Aspirin

 summary:

 The procedure for the synthesis of L-serine, L-threonine, and L-hydroxyproline esters will be outlined in the Synthesis Sequence section, and these schemes are illustrative examples for other amino acids. The complete process and analytical data will be provided in the experimental section. Generally acetylsalicyloyl chloride (in batches, 10 g-25 g) bound with N-benzyloxy / benzyl ester protected amino acids in the presence of pyridine. Once the reactions were complete (at room temperature, 24 to 48 hours), the mixture was poured into ice cold 2N hydrochloric acid. The dichloromethane layer was then washed with sodium bicarbonate, water and brine. After drying over sodium sulfate, filtration and concentration, the amino acid esters of acetylsalicylic acid, which have not yet been processed and purified, were purified by flash chromatography on silica gel. This process yielded the amino acid esters of protected acetylsalicylic acid in the yield range of 68% to 95%. The protecting groups were removed by hydrogenation (20 psi hydrogen) in the presence of 10% palladium on carbon. The yield of the deprotection step was generally in the range of 70-90%. The amino acid esters of acetylsalicylic acid were extracted with water and the palladium catalyst was concentrated and dried. The final compounds were washed with solvent (water, dioxane, acetonitrile, and / or dichloromethane) until pure and dried under high pressure vacuum conditions until a constant weight was obtained.

Synthetic sequence:

One. SPIB00102

2. SPIB00101

3. SPIB00103

Synthesis of L-serine, L-threonine, and L -hydroxyproline esters of acetylsalicylic acid : a) pyridine, CH ₂ Cl ₂ ; b) 10% Pd / C, EtOH, EtOAc.

Part of the experiment:

Synthesis of SPIB00101, SPIB00102 and SPIB00103 was performed in single or two batches. Reagents mentioned in the experimental section were purchased with the highest purity available from Lancaster, Sigma, Aldrich, Acros or Bachem, except for solvents purchased from either Fisher Scientific or Mallinkrodt.

1) SPIB00102: 2-O-acetylsalicylic acid (2S, 3R)-(-)-threonine ester

A mixture of N-carbobenzyloxy-L-threonine benzyl ester (Z-Thr-OBzl, 21.77 g, 63.40 mmole) and pyridine (25 mL) in anhydrous dichloromethane (500 mL) in an ice water bath Cooled. Acetylsalicyloyl chloride (17.63 g, 88.76 mmole) was added and the mixture was then warmed to room temperature and stirred overnight. After 24 hours, the mixture was poured into ice cold 2N hydrochloric acid (400 mL). After mixing, the layers were separated and the dichloromethane layer was washed with water (500 mL), saturated sodium bicarbonate (500 mL), water (500 mL), brine (500 mL). Dry over sodium sulfate (25 g). After filtration and concentration under reduced pressure, dried under high pressure vacuum conditions, the remaining yellow oil (35.43 g) was dissolved in hexane / ethyl acetate (3: 3) on silica gel (300 g, 0.035-0.070 mm, 6 nm pore diameter). Purification by flash chromatography eluting with 1). After the product containing fractions were concentrated under reduced pressure conditions and then dried under high pressure vacuum conditions until a constant weight was obtained, the procedure was followed by the protected acetylsalicylic-L-threonine ester SPIB0010201 (28.1 g, 88%). Yield) was produced as a colorless oil.

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 7.74 (1H, d, J = 7.5 Hz), 7.51 (1H, dt, J = 7.5, 1.5 Hz), 7.34-7.17 (1H, m), 7.06 (1H, d, J = 7.2 Hz), 5.62 (2H, m), 5.13 (4H, m), 4.65 (1H, dd, J = 9.6, 2.4 Hz), 2.29 (3H, s), 1.38 (3H , d, J = 6.6 Hz).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 169.35, 169.22, 162.73, 156.26, 150.41, 135.79, 134.67, 133.77, 131.24, 128.35, 128.24, 128.08, 127.95, 125.78, 123.51, 122.61, 71.22, 67.26, 67.72 , 57.64, 20.98, 16.88.

Protected acetylsalicylic-L-threonine ester SPIB0010201 (14.50 g, 28.68 mmole) was dissolved in ethanol (100 mL) and ethyl acetate (100 mL) at room temperature and under nitrogen atmosphere, 10% palladium (3.0 g) on carbon , 50% wet) was poured into a Parr bottle. The nitrogen atmosphere was replaced with hydrogen gas (20 psi). After stirring for 20 hours, the palladium catalyst was removed by filtration through celite. The remaining white solids (palladium / celite and product) were washed with water (600x4 mL) until the product was removed. The ethanol and water fractions were concentrated under reduced pressure at room temperature. The remaining solids were washed with water (20 mL) and dioxane (20 mL) for 48 hours. After filtration, the remaining white solids were dried (16 h) at room temperature until constant weight was obtained under high pressure vacuum conditions. This experiment produced acetylsalicylic-L-threonine ester, SPIB00102 (4.40 g, 55% yield) as a white solid,

¹ H NMR (300 MHz, D ₂ O-DCl): δ = 8.00 (1H, dd, J = 7.8, 1.5 Hz), 7.74 (1H, dt, J = 7.8, 1.5 Hz), 7.47 (1H, dt, J = 7.8, 1.5 Hz), 7.27 (1H, dd, J = 7.8, 1.5 Hz), 5.76 (1H, dq, J = 6.9, 3.0 Hz), 4.49 (1H, d, J = 3.0 Hz), 2.39 ( 3H, s), 1.55 (3H, d, J = 6.9 Hz).

¹³ C NMR (75 MHz, D20-DCl): δ = 173.03, 168.84, 163.97, 149.56, 135.32, 131.26, 126.85, 123.48, 121.49, 69.16, 56.36, 20.45, 15.86.

HPLC analysis: 98.7% purity; rt = 6.233 min; Luna C18 5u column (sn 167917-13); 4.6 x 250 mm; 254 nm; 35% MeOH / 65% TFA (0.1%) pH = 1.95; 35 C; 20 ul inj .; 1 ml / min; The sample was dissolved in the mobile phase with one drop of phosphoric acid.

CHN analysis: calc .: C 55.51, H 5.38, N 4.98; found: C 55.37, H 5.40, N 5.03.

Melting Point: 153.5 ° C (dec.)

2) SPIB00101: 2-O-acetylsalicylic acid (2S)-(+)-serine ester

A mixture of N-carbobenzyloxy-L-serine benzyl ester (Z-Ser-OBzl, 23.17 g, 70.34 mmole) and pyridine (30 mL) in anhydrous dichloromethane (500 ml) in an ice water bath Cooled. Acetylsalicyloyl chloride (21.07 g, 106.1 mmole) was added and the mixture was then warmed to room temperature and stirred over 2 hours. After 48 hours, the mixture was poured into ice cold 2N hydrochloric acid (400 mL). After mixing, the layers were separated and the dichloromethane layer was washed with water (500 mL), saturated sodium bicarbonate solution (500 mL), water (500 mL), brine (500 mL). Dry over sodium sulfate (25 g). After filtration and concentration under reduced pressure, dried under high pressure vacuum conditions, the remaining yellow oil (47.19 g) was purified on hexane / ethyl acetate (200 g, 0.035-0.070 mm, 6 nm pore diameter) on silica gel. Purification by flash chromatography eluting with 3: 1). After the product containing fractions was concentrated under reduced pressure conditions and then dried under high pressure vacuum conditions until a constant weight was obtained, the procedure was followed by the protected acetylsalicylic-L-serine esters SPIB0010101 (32.97 g, 95 % Yield) was produced as a white solid.

¹ H NMR (300 MHz, CD13): δ = 7.74 (1H, d, J = 7.8 Hz), 7.55 (1H, dt, J = 7.8, 1.5 Hz), 7.33-7.21 (11H, m), 7.08 (1H) , d, J = 7.5 Hz), 5.68 (1H, d, J = 8.4 Hz), 5.20 (2H, s), 5.12 (2H, s), 4.77 (1H, m), 4.66 (1H, dd, J = 11.4, 3.3 Hz), 4.57 (1H, doublet of doublets, J = 11.4, 3.3 Hz), 2.30 (3H, s).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 169.45, 169.09, 163.68, 163.35, 155.57, 150.77, 135.87, 134.75, 134.07, 131.44, 128.50, 128.43, 128.27, 128.14, 128.04, 125.92, 123.71, 122.18, 122.18 , 67.27, 64.63, 53.55, 21.03.

Protected acetylsalicylic-L-serine esters SPIB0010101 (21.0 g, 42.7 mmole) were dissolved in ethanol (100 mL) and ethyl acetate (100 mL) at room temperature, and under nitrogen atmosphere, 10% palladium (4.20) on carbon. g, 50% wet) was poured into a Parr bottle. The nitrogen atmosphere was replaced with hydrogen gas (20 psi). After stirring for 5 hours, an additional 10% palladium catalyst (4.26 g) was added and its hydrogen atmosphere (20 psi) was reduced. After an additional 20 minutes of stirring at room temperature, the palladium catalyst was removed by filtration through celite. The remaining white solids (palladium / celite and product) were washed with water (1500x2 mL) until the product was removed. The ethanol and water fractions were concentrated under reduced pressure at room temperature. The remaining solids (7.17 g) were dissolved in DIUF water (4.3 L) and filtered through celite to remove insoluble matter. It was then concentrated under high pressure vacuum conditions at room temperature. The white solid was then washed overnight with 1,4-dioxane (100 mL) and DIUF water (50 mL). After 24 hours, the solids were filtered and dried under high pressure vacuum until constant weight was obtained (24 hours). This experiment produced acetylsalicylic-L-serine ester SPIB00101 (6.17 g, 54% yield) as a white solid.

¹ H NMR (300 MHz, D ₂ O-DCl): δ = 8.05 (1H, dd, J = 7.8, 1.5 Hz), 7.75 (1H, dt, J = 7.8, 1.5 Hz), 7.47 (1H, dt, J = 7.8, 0.9 Hz), 7.27 (1H, dd, J = 7.8, 0.9 Hz), 4.87 (1H, dd, J = 12.6, 4.2 Hz), 4.79 (1H, dd, J = 12. 6,3. 0 Hz), 4.62 (1H, doublet of doublets, J = 4.2,3.0 Hz), 2.39 (3H, s).

¹³ C NMR (75 MHz, D ₂ 0-DCl): δ = 173.01, 168.58, 164.54, 149.72, 135.39, 131.59, 126.87, 123.62, 121.15, 62.38, 52.05, 20.44.

HPLC analysis: 98.1% purity; rt = 5.839 min .; 65% TFA (0.1%) / 35% methanol; 1 mL / min; 35 C; Luna C18, 3u column (SN 184225-37), 4.6 × 250 mm; 22 ul injection; DAD1B, Sir = 240, 4Ref = 550,100.

CHN analysis: calc .: C 53.93, H 4.90, N 5.24; found: C 54.02, H 5.00, N 5.23.

Melting Point: 147.0 ° C (dec.)

3) SPIB00103: 2-O-acetylsalicylic acid (2S, 4R) -4-hydroxyproline ester

A mixture of N-carbobenzyloxy-L-hydroxyproline benzyl ester (Z-Ser-OBzl, 21.5 g, 60.5 mmole) ¹ and pyridine (25 mL) in anhydrous dichloromethane (500 mL) Cooled in a water bath. Acetylsalicyloyl chloride (13.2 g, 66.6 mmole) was added and the mixture was then warmed to room temperature and stirred overnight. After 24 hours, additional acetylsalicyloyl chloride (5.0 g, 25.2 mmole) was added and the mixture was allowed to stir overnight. After 48 hours, the mixture was poured into 1N hydrochloric acid (500 mL) cooled with ice. After mixing, the layers were separated and the dichloromethane layer was washed with water (500 mL), saturated sodium bicarbonate solution (500 mL), water (500 mL), brine (500 mL). Dry over sodium sulfate (25 g). After filtration and concentration under reduced pressure, dried under high pressure vacuum conditions, the remaining yellow oil (40.7 g) was purified on heptane / ethyl acetate (460 g, 0.035-0.070 mm, 6 nm pore diameter) on silica gel (460 g). Purification by flash chromatography eluting with 3: 1). After the product containing fractions were concentrated under reduced pressure conditions and then dried under high pressure vacuum conditions until a constant weight was obtained, the procedure was followed by the protected acetylsalicylic-L-hydroxyproline ester SPIB0010301 (21.31 g, 68% yield) as a colorless oil.

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 7.92 (1H, d, J = 7.8 Hz), 7.56 (1H, t, J = 7.8 Hz), 7.34-7.21 (1OH, m), 7.09 (1H, d, J = 7.8 Hz), 5.48 (1H, s), 5.21 (2H, m), 5.03 (2H, d, J = 15 Hz), 4.57 (1H, m), 3.85 (2H, m), 2.53 ( 1 H, m), 2.28 (4 H, m).

¹³ C NMR (75 MHz, CDCl ₃ ): δ = 171.72, 171.49, 169.25, 163.47, 163.30, 154.52, 153.93, 150.54, 136.05, 135.94, 135.21, 135.00, 134.17, 134.12, 128.43, 128.32, 128.28, 128.20, 128.20, 128. , 127.98, 127.94, 127.79, 125.89, 123.70, 122.46, 122.38, 73.24, 72.59, 67.33, 67.11, 66.97, 58.02, 57.69, 52.47, 52.15, 36.74, 35.65, 20.90.

Protected acetylsalicylic-L-hydroxyproline esters SPIB0010301 (10.6 g, 20.5 mmole) were dissolved in ethanol (75 mL) and ethyl acetate (75 mL) at room temperature, and under nitrogen atmosphere, 10% on carbon It was poured into a Parr bottle containing palladium (3.0 g, 50% wet). The nitrogen atmosphere was replaced with hydrogen gas (20 psi). After stirring at room temperature for 17 hours, the reaction mixture was washed with water (500 mL) for 2 hours. The organic acid layer (top) was removed through a pipette and the aqueous layer was filtered through celite. The water fraction was concentrated under reduced pressure at room temperature. The remaining solids (6.71 g) were then washed overnight with anhydrous dichloromethane (35 ml). After 24 hours, the solid was filtered and dried under high pressure vacuum conditions until constant weight was obtained (24 hours). This experiment produced acetylsalicylic-L-hydroxyproline ester, SPIB00301 (2.87 g, 47.7% yield) as a white solid.

¹ H NMR (300 MHz, D ₂ O-DCl): δ = 8. 09 (1H, d, J = 7.5 Hz), 7.75 (1H, t, J = 7.5 Hz), 7.48 (1H, t, J = 7.5 Hz), 7.28 (1H, d, J = 7.5 Hz), 5.69 (1H, m), 4.76 (1H, t, J = 7.5 Hz), 3.86 (1H, dd, J = 13.5, 3.9 Hz) , 3.74 (1H, d, J = 13.5 Hz), 2.81 (1H, dd, J = 15.0, 7.5 Hz), 2.60 (1H, m), 2.40 (3H, s).

¹³ C NMR (75 MHz, D ₂ O-DCl): δ = 173.13, 170.25, 164.31, 149.65, 135.36, 131.54, 126.87, 123.54, 121.37, 73.86, 58.34, 50.95, 34.38, 20.48.

HPLC analysis: 98.3% purity; rt = 7.201 min. ; 65% TFA (0.1%) / 35% methanol; 1 mL / min; 35 C; Luna C18, 3u column (SN 184225-37), 4.6 × 250 mm; 22 ul scan; DAD1B, Sig = 240, 4Ref = 550,100.

CHN analysis: calc .: C 57.34, H 5.16, N 4.78; found: C 57.09, H 5.23, N 4. 91.

Melting Point: 162 ℃ (dec.)

Gastrointestinal mucosal irritating potential of L-serine, L-threonine, and L -hydroxyproline esters of acetylsalicylic acid compared to acetylsalicylic acid :-

This study is responsible for gastrointestinal mucosal irritations / lesions in fasting male albino rats (Wistar strain) in new formulations of aspirin (L-serine, L-threonine, and L-hydroxyproline esters of acetylsalicylic acid). Was performed to determine its relative potential. Aspirin served as a reference control.

Aspirin and other types of new formulations of the reference control and aspirin in male albino rats using 0.5% (w / v) Carboxymethylcellulose (CMC) in phosphate buffer (pH 2.6) as the carrier. (Swiss strain) was administered via a feeding gavage. This study was performed at two doses, 100 mg and 200 mg / kg body weight, with vehicle control. Five animals were used at each dose. All doses were expressed in molar equivalents of aspirin. The molar equivalents as well as the dosages used are given below.

Formulation: molar equivalent Formulation Molar equivalent L-serine ester of acetylsalicylic acid 1.483 units is equivalent to 1 unit of aspirin. L-hydroxyproline ester of acetylsalicylic acid 1.628 units is equivalent to 1 unit of aspirin. L-threonine ester of acetylsalicylic acid 1.561 units is equivalent to 1 unit of ibuprofen.

Test item: Group: Dose (mg per kg) [For acetylsalicylic acid]: Equivalent weight of test item [mg] Test Items group Dose (mg per kg) [in terms of acetylsalicylic acid] Equivalent Weight of Test Item [mg] Vehicle control Vehicle control 0.0 - L-serine ester of acetylsalicylic acid Trial group 1 100.0 148.3 Test group 2 200.0 296.6 L-Haxyhydroxyproline Esters of Acetylsalicylic Acid Trial group 1 100.0 162.8 Test group 2 200.0 325.6 L-threonine ester of acetylsalicylic acid Trial group 1 100.0 156.1 Test group 2 200.0 312.2 Reference Control Acetylsalicylic Acid Trial group 1 100.0 100.0 Test group 2 200.0 200.0

Mice are fasted on a cycle of 18 to 22 hours prior to administration. The test item was administered as a single dose via nutrient gavage. For three hours after drug administration, the animals were allowed to die humanly through inhalation of carbon dioxide gas. The stomach was removed and the following were observed.

 ● amount of mucosal exudate,

 ● redness and thickening of the stomach wall

 The location of the bleeding (local or sporadic), the nature of the bleeding according to its size (petechial or orecchymotic)

 ● perforated

The observations of gastrointestinal mucosa irritation in various groups of animals are summarized below.

Test Item: Group: Dose mg / kg (as acetylsalicylic acid): Observation Test Items group Dose mg per kg (relative to acetylsalicylic acid) observe Vehicle control Vehicle control 0.0 None of the animals showed any evidence of gastric mucosal irritation L-serine ester of acetylsalicylic acid Trial group 1 100.0 None of the animals showed any evidence of gastric mucosal irritation Test group 2 200.0 None of the animals showed any evidence of gastric mucosal irritation -L-hydroxyproline ester of acetylsalicylic acid Trial group 1 100.0 None of the animals showed any evidence of gastric mucosal irritation Test group 2 200.0 None of the animals showed any evidence of gastric mucosal irritation L-threonine ester of acetylsalicylic acid Trial group 1 100.0 None of the animals showed any evidence of gastric mucosal irritation Test group 2 200.0 None of the animals showed any evidence of gastric mucosal irritation Reference control (acetylsalicylic acid) Trial group 1 100.0 None of the animals showed any evidence of gastric mucosal irritation Test group 2 200.0 5 animals administered
Both showed gastric mucosal irritation

As a result, it was observed that the L-serine, L-threonine, and L-hydroxyproline esters of acetylsalicylic acid induced some evidence of irritation to the gastrointestinal mucosa at two doses, 100 mg and 200 mg / kg body weight. . In contrast, aspirin (acetylsalicylic acid) caused irritation to the gastrointestinal mucosa in male fasting albino rats at a dose of 200 mg / kg. However, the 100 mg / kg dose of aspirin did not cause irritation to any gastrointestinal mucosa in rat males. None of the animals in the other experimental groups showed any clinical signs of toxicity during the three hour observation cycle.

Acetylsalicylic acid L- serine one of acetylsalicylic acid compared to the clotting time which is estimated at a time after administration in mice, L- threonine, and the effects of the L- hydroxyproline ester

 Observation of Blood Coagulation Time

Low, moderate and high dose groups with mean clotting time (MCT), vehicle control and positive for other formulations estimated at one hour after administration in animals. Positive control groups are presented below (Table 14):

Summary of mean clotting time (± S.D.) In trace-L-serine, L-threonine, and L-hydroxyproline esters of acetylsalicylic acid and aspirin (positive control): Low Dose: Medium Dose: High Dose Low dose Medium dose High dose Vehicle control 4.9 ± 1.10 L-serine ester of acetylsalicylic acid 5.7 ± 1.34 6.8 ± 1.48 6.9 ± 1.37 -L-hydroxyproline ester of acetylsalicylic acid 6.1 ± 1.10 5.7 ± 0.82 7.5 ± 1.18 L-threonine ester of acetylsalicylic acid 5.2 ± 1.14 5.6 ± 0.84 7.4 ± 0.97 Positive control (acetylsalicylic acid) 6.2 ± 1.40 8.1 ± 1.97 9.8 ± 1.32

3-6 show group mean data of animals regarding dose relationship + mean blood clotting time in minutes for L-series esters of aspirin and controls.

The statistical analysis showed a significant improvement with a significant level of 5% in efficacy for high and moderate doses when compared to the vehicle control (FIG. 7).

4 shows group mean data in animals. It provides a dose response relationship to the mean coagulation time (MCT) in minutes with respect to the L-hydroxyproline ester of asperin. The statistical analysis of FIG. 4 showed a significant improvement, with a significant level of 5% in efficacy for high and moderate doses when compared to the vehicle control (FIG. 6).

Figure 5 shows the dose response relationship to mean blood clotting time (MCT) in minutes of the L-threonine ester of acetylsalicylic acid. The statistical analysis showed a significant improvement, with a significant level of 5% in efficacy for high and moderate doses when compared to the vehicle control.

6 shows the dose response relationship to mean blood clotting time for acetylsalicylic acid. The statistical analysis showed a significant improvement, with a significant level of 5% in efficacy for moderate and high doses, when compared to the vehicle control. The dose response effect was statistically significant and was clearly apparent (FIG. 7).

conclusion

The present invention was performed to evaluate the efficacy of new formulations of aspirin using blood clotting time as an index in albino rats. Aspirin served as a positive control.

This study was conducted at three doses with the new formulations and the positive control along with the vehicle control.

Dose

Doses for the main study were selected based on the dose ranges obtained by finding experiments with acetylsalicylic acid. All doses were expressed in molar equivalents of aspirin and the doses used in the main experiments for the different formulations and the positive control are the same and are presented below.

Test Item: Low Dose (mg / kg): Medium Dose (mg / kg): High Dose (mg / kg) Test Items Low dose (mg / kg) Medium dose (mg / kg) High dose (mg / kg) L-serine ester of acetylsalicylic acid 1.0 4.0 10.0 -L-hydroxyproline ester of acetylsalicylic acid 1.0 4.0 10.0 L-threonine ester of acetylsalicylic acid 1.0 4.0 10.0 Aspirin (positive control) 1.0 4.0 10.0

Efficacy (blood coagulation time)

Efficacy in terms of time required for coagulation time at different dosages of acetylsalicylic acid and different dosages estimated at one hour after administration in animals and at different dosages of high doses Is presented in

Low Dose: Medium Dose: High Dose Low dose Medium dose High dose Vehicle control 4.9 ± 1.10 L-serine ester of acetylsalicylic acid 5.7 ± 1.34 6.8 ± 1.48 6.9 ± 1.37 -L-hydroxyproline ester of acetylsalicylic acid 6.1 ± 1.10 5.7 ± 0.82 7.5 ± 1.18 L-threonine ester of acetylsalicylic acid 5.2 ± 1.14 5.6 ± 0.84 7.4 ± 0.97 Positive control 6.2 ± 1.40 8.1 ± 1.97 9.8 ± 1.32

L-serine, L-threonine, and L-hydroxyproline esters of acetylsalicylic acid show significant levels of acetylsalicylic acid for the blood coagulation time observed after one hour, but are not gastrointestinal irritant at all concentrations when compared to acetylsalicylic acid. Excellent in terms of

After administration in mice of acetylsalicylic acid compared to acetylsalicylic acid on the blood coagulation time which is estimated at two times L- serine, L- threonine, and the effects of the L- hydroxyproline ester

The present invention utilizes the blood coagulation time estimated at two hours (± 10 minutes) after administration in albino rats to determine the efficacy of L-serine, L-threonine, and L-hydroxyproline esters of acetylsalicylic acid compared to acetylsalicylic acid. It was performed to evaluate as an index. Aspirin served as a positive control. Male albino rats were exposed to new formulations of aspirin and three aspirin at one dose level of 20 mg / kg body weight. No vehicle control was used. All doses were expressed in molar equivalents of aspirin and the doses used in the main experiments for the different formulations and the positive control are the same and are presented below.

Test item: Dose of acetylsalicylic acid (mg / kg) Test Items Dose of acetylsalicylic acid (mg / kg) L-serine ester of acetylsalicylic acid 20.0 -L-hydroxyproline ester of acetylsalicylic acid 20.0 L-threonine ester of acetylsalicylic acid 20.0 Aspirin (positive control) 20.0

Efficacy (blood coagulation time)

The efficacy in terms of time required for coagulation time at a concentration of 20 mg / kg body weight for different formulations and aspirin (positive control) is shown below.

Observation of Blood Coagulation Time

Data from the mean blood clotting time estimated at 2 hours (± 10 minutes) after administration in the animals, at concentrations of 20 mg / kg body weight, different formulations, vehicle control and positive controls groups are given below.

Summary of mean clotting time (± S.D.) in trace amounts of L-serine, L-threonine, and L-hydroxyproline of acetylsalicylic acid compared to acetylsalicylic acid Dose (20mg / kg) L-serine ester of acetylsalicylic acid 3.8 ± 0.92 -L-hydroxyproline ester of acetylsalicylic acid 4.2 ± 1.32 L-threonine ester of acetylsalicylic acid 5.3 ± 1.06 Positive control (acetylsalicylic acid) 5.4 ± 1.17

L-serine, L-threonine, and L-hydroxyproline esters of acetylsalicylic acid have been shown to be effective at blood coagulation times.

In conclusion, when estimated at two hours after dosing, based on the time required for blood to coagulate (blood coagulation time), the amino acid prodrugs were observed to be efficacious. However, it has been found that the L-threonine ester of acetylsalicylic acid has a relatively better efficacy than the other two formulations.

As shown in FIG. 7, statistical analysis showed that L-threonine, and L-hydroxyproline esters of acetylsalicylic acid are as effective as acetylsalicylic acid. With respect to the positive control for the mean coagulation time observed after two hours, there was no significant difference in the 5% significance level for the L-hydroxyproline ester of acetylsalicylic acid and the L-threonine ester of acetylsalicylic acid. However, when associated with gastrointestinal irritant potential, L-serine, L-threonine, and L-hydroxyproline esters of acetylsalicylic acid are much better.

There are many screening experiments for determining the use of the prodrugs invented in accordance with the disclosed methods. These experiments include screening methods for both in vitro and in vivo experiments.

In vitro methods include acid / base hydrolysis of prodrugs, hydrolysis in porcine pancreas, hydrolysis in rat intestinal fluid, hydrolysis in human gastric juice, hydrolysis in human intestinal fluid, and hydrolysis in human plasma. These tests are described in Simmons, DM, Chandran, VR and Portmann, GA, Danazol Amino Acid Prodrugs: In Vitro and In Situ Biopharmaceutical Evaluation, Drug Development and Industrial Pharmacy, Vol 21, Issue 6, Page 687,1995. The contents of which are incorporated by reference.

The compounds of the present invention are effective in treating diseases or disorders in which nonsteroidal anti-inflammatory drugs are normally used. The prodrugs disclosed herein enhance the therapeutic advantages of nonsteroidal anti-inflammatory drugs by reducing or eliminating the biopharmaceutical and pharmacokinetic barriers that are modified in the human body and associated with each of them to release the active compound. . However, it should be noted that these prodrugs themselves will have sufficient functionality without releasing any effective agent in mammals. Since the prodrugs are more soluble in water, they need to be associated with ibuprofen or other nonsteroidal anti-inflammatory drugs, carrier carriers such as alcohol or castor oil, which may be toxic or may produce unwanted additional reactions. There is no. In addition, oral formulations containing prodrugs of nonsteroidal anti-inflammatory drugs are absorbed into the blood and are quite effective.

Thus, the prodrugs of the present invention enhance the therapeutic advantages of nonsteroidal anti-inflammatory drugs by removing the biopharmaceutical and pharmacokinetic barriers of existing drugs.

Furthermore, the prodrugs are easily synthesized in high yields, using materials that are easy to use and are currently available.

IV . Of acetaminophen  Proline derivatives

summary:

The procedure for the synthesis of the L-proline ester of acetaminophen will be outlined in the Synthesis Sequence section. These syntheses are illustrative examples. The complete process and analytical data will be provided in the experimental section. Acetaminophen (10 g) was bound to Boc-L-proline by EDC in the presence of a catalytic dose of DMAP. Once the reactions were complete (3 hours at room temperature), the solution was washed with water. After drying over sodium sulfate, filtration and concentration, the amino acid esters of acetaminophen that have not yet been processed and purified were purified by flash chromatography on silica gel. The process produced 72% of the L-proline ester of protected acetaminophen. The protecting group was removed by dissolving the ester in dichloromethane and passing the hydrogen chloride through the solution at room temperature. After the filtration process, the final salt was stirred in tetrahydrofuran until it was pure. After filtration and drying at 90 ° C. for 4 hours under high vacuum, the yield for the deprotection step was 91.4%.

Compound sequence

Synthesis of L-proline ester of acetaminophen : a) EDC, DMAP, CH ₂ Cl ₂ ; b) HCl (g), CH ₂ Cl ₂ .

Part of the experiment:

Synthesis of SPI0014 was performed in a single batch. The materials mentioned in the experimental section were purchased with the highest purity available from Lancaster, Sigma-Aldrich or Acros, or Bachem, with the exception of solvents purchased from either Fisher Scientific or Mallinkrodt.

SPI0014 : Pyrrolidine-2 (S) -carboxylic acid 4-acetylamino-phenyl ester, hydrochloride

A mixture of Boc-L-proline (14.39 g, 68.80 mmole), acetaminophen (10.02 g, 66.28 mmole), EDC (12.9 g, 67.29 mmole) and DMAP (1.10 g, 9.0 mmole) in anhydrous dichloromethane (100 mL) After stirring for 3 hours at room temperature under silver argon atmosphere, water (120 mL) was added. After mixing for 5 minutes, the layers were separated and the dichloromethane layer was washed again with water (120 mL). Dry over sodium sulfate (5 g). After filtration, concentration under reduced pressure, and drying under high pressure vacuum conditions, the remaining oil (24.10 g) was deposited on silica gel (100 g, 0.035-0.070 mm, 6 nm pore diameter), Purification by column chromatography eluting with hexane / ethyl acetate (1: 2). The product containing fractions was concentrated under reduced pressure conditions and after drying under high pressure vacuum conditions until the weight was constant, the experiment showed that the L-proline ester of protected acetaminophen as a white solid (foam) was used as SPI001401 ( 16.71 g, 72.3% yield).

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 8.83 (1/2 H, s), 8.70 (1/2 H, s), 7.58 (1/2 H, d, J = 7.5 Hz), 7.46 ( 1/2 H, d, J = 7.5 Hz), 6.96 (2H, m), 4.47 (1H, m), 3.59-3. 45 (2H, m), 2.36 (1H, m), 2.17-1. 90 (6 H, m), 1.46 (9 H, m).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 171.91, 171.75, 169.02, 154.44, 153.78, 146.36, 146.21, 121.44, 121.23, 120.82, 80.41, 80.17, 59.16, 46.78, 46.55, 31.06, 30.11, 28.50, 24.57, 28.50 , 24.28, 23.78.

Protected acetaminophen-L-proline ester SPI001401 (16.60 g, 47.64 mmole) was dissolved in dichloromethane (400 mL) and hydrogen chloride was allowed to pass through the solution for 2 hours at room temperature. The remaining solids were left to stabilize (for 1 hour). The dichloromethane was carefully removed from the white precipitate. Tetrahydrofuran (200 mL) was added to the precipitate and the mixture was stirred for 2 hours under argon atmosphere. After the filtration process, the remaining white solid was dried under high vacuum at 90 ° C. until the weight of the product was constant (4 hours). This experiment produced the L-proline ester of acetaminophen, hydrochloride SPI0014 (12.4 g, 91.4% yield) as a white solid.

¹ H NMR (300 MHz, CDCl ₃ -DMSO): δ = 10.41 (1H, br s), 10.26 (1H, s), 9.55 (1H, br s), 7.70 (2H, d, J = 9 Hz), 7.12 (2H, d, J = 9 Hz), 4.66 (t, 1H, J = 8. 4 Hz), 3.33 (2H, m), 2.43 (1H, m), 2.28 (1H, m), 2. 08 (s, 3 H), 2.04 (2 H, m).

¹³ C NMR (75 MHz, CDCl ₃ -DMSO): δ = 168.08, 167.25, 144.55, 137.40, 121.12, 119.64, 58.53, 45.33, 27.74, 23.86, 23.08.

HPLC analysis: 99.45% purity; rt-5. 733 min; Luna C18 5u column (sn 167917-13); 4.6 x 250 mm; 254 nm; 15% MeOH / 85% hexanes sulfonate buffer (110mMol, pH = 6); 35 C; 20 ul inj .; lml / min; 5 mg / mL specimen size.

CHN analysis: calc .: C 54.84, H 6.02, N 9.84; found: C 54.66, H 5.98, N 9.65.

Melting Point: 221-222 ℃

V. Cyclosporin  Amino acid derivatives of A

 Macrocyclic immunosuppresants are structurally distinctive, cyclic, poly, N-methylated and uncapapted, and similar semi-synthetic macrolides. macrolide) and include a class that commonly possesses pharmacological, in particular immunosuppressive, anti-inflammatory and / or anti-parasitic activity. The first cyclosporine isolated was a naturally occurring fungal metabolite, cyclosporin or cyclosporine, also known as cyclosporin A, which has the formula

Where MeBmt represents the N-methyl- (4R) -4-but-2E-en-1-yl-4-methyl- (L) threonyl moiety of the formula:

Where -x-y- is CH = CH- (trans). Other similar products include sirolimus (b), tacrolimus (c), and pimecrolimus (d), which have the following structures:

Thus the class covered by cyclosporins is actually very large, for example, [Thr] ^2- , [Val] ^2- , [Nva] ² -and [Nva] ^2- [Nva] ⁵ -cyclosporine (also , Respectively known as cyclosporin C, D, G and M), [Dihydrop-MeBmt] ^1- [Val] ² -cyclosporin (also known as dihydro-cyclosporin D), [(D) Ser] ⁸ -cyclosporin, [MeIle] ¹¹ -cyclosporin, [(D) MeVal] ¹¹ -cyclosporin (also known as cyclosporin H), [MeAla] ⁶ -cyclosporin, [(D) Pro] ³ -cyclo Sporeins and the like.

According to the conventional nomenclature for cyclosporins, they are defined throughout the specification and claims referring to the structure of cyclosporin (ie cyclosporin A). This is different from the ionic cyclosporine (eg, "[(D) Pro] ³ ) present, to indicate that the subject cyclosporin has-(D) Pro- rather than -Sar- residue at the 3-position. The amino acid residues present are first indicated and then named by applying the term cyclosporin to characterize the remaining residues identical to those present in cyclosporin A.

As used herein, the term “cyclosporins” refers to various types of cyclosporins, where xy at the MeBmt residue has cis or trans CH═CH, wherein xy at positions 2-11 of cyclosporin A Also included are derivatives in which one or more amino acids in are substituted with other amino acids. However, it is preferable that at most two amino acids are substituted in the formula of cyclosporin and more preferably at most one amino acid is substituted with an amino acid.

Additionally, amino acid residues may be abbreviated, for example, with -Ala-,-MeVal- and -aAbu- and (L) -batch by conventional usage, and otherwise, for example, " -(D) Ala- "to be understood as in the case. The residue abbreviations preceded by "Me" as in the case of "-MeLeu-" refer to -N-methylated residues. The residues of the individual cyclosporine molecules are numbered clockwise in this technique, starting with the residues -MeBmt-, dihydro-MeBmt-, etc ... at position 1. The same sequence of numbers is used throughout the specification and claims.

Because of their unique pharmaceutical potential, macrocyclic immunosuppressants have attracted considerable media attention. The term "macrocyclic immunosuppressants" includes various natural derivatives and semi-synthetic derivatives of cyclosporine and other macrolides such as sirolimus, tacrolimus and pimecrolimus. The primary area of clinical investigation of these drugs was such as immunosuppressive drugs, in particular the application of the drugs to recipients of organ transplants, for example, heart, lung, combined heart-lung Liver, kidney, pancreas, bone marrow, skin and cornea transplants, and especially allogenic organ transplants.

Macrocyclic molecular immunosuppressants are also used for various autoimmune diseases and inflammatory diseases and especially arthritis (eg, rheumatoid arthritis, arthritis, and arthritis chronica progredient deformans). And inflammatory diseases with etiology involving autoimmune elements such as rheumatic diseases. Certain cyclosporins for which cyclosporine therapy has been proposed or applied include autologous hemologic dysfunctions (eg, hemolytic anemia, aplastic anemia, pure red cell anemia, and idiopathic platelets). Idiopathic thrombocytopaenia, systemic lupus erythematosus, polychondritis, sclerodeoma, Wegener granulamatosis, dermatomyositis, chronic active hepatitis , Myasthenia gravis, psoriasis, Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease-for example, ulcerative colitis and Crohn's disease and endocrine eye Endocrine opthalmopathy Graves disease, sarcoidosis, multiple sclerosis, primary biliary cirrhosis (pr imary billiary cirrhosis, juvenile diabetes (diabetes mellitus typeI), uveitis (anterior and posterior), keratoconjunctivitis sicca and vernal keratoconjunctivitis, mesenchymal pulmonary fibrosis ), Psoriatic arthritis, atopic dermatitis, and glomerulonephritis (with or without nephrotic syndrome, for example, idiopathic nephritic syndrome or minimal change) Nephropathy (minimal change nephropathy).

Furthermore, macromolecular immunosuppressants also have applicability as antiparasitic, especially as anti-protozoal agents and in malaria, coccidiomycosis and schistosomiasis. (schistomsomiasis) has been suggested to be useful. More recently, macrocyclic immunosuppressants have been known to be useful as agents to reverse or eliminate anti-neoplastic agent resistance contumors, and the like.

Despite the major contributions that macrocyclic immunosuppressants are making, reported unwanted additional reactions; In particular, neurotoxic reactions are not only clear and serious obstacles to the widespread use or application of the medicament, but also difficulties encountered while providing more effective and convenient methods of administration (eg galenic formulations, eg, Oral dosage forms for the patient, as well as providing adequate bioavailability and enabling dosing at an appropriate and controlled rate).

In addition, the macrocyclic molecular immunosuppressants mentioned above are characteristically very hydrophobic and easily settle even where very little water is present. E.g. This is true even if it comes into contact with the human body (for example, gastric juice). Thus, for example. In terms of form and taste, it is extremely difficult to provide oral formulations that are easy to administer to a patient, are stable when stored, and that can be administered as usual in order to provide appropriate and patient-controlled dosages.

Presented liquid formulations, for example. Those for oral administration of macrocyclic immunosuppressants, to date, have been based primarily on the use of ethanol and oils or similar excipients as carrier medium. Thus, currently commercially available drink-solutions of macrocyclic immunosuppressants use ethanol and olive oil or corn oil as carrier media, for example in connection with solvent systems comprising ethanol and LABRIFIL and equivalent excipients as carrier media. do. For example, commercially available macrocyclic molecular immunosuppressant drink solutions use ethanol and olive or corn oil as carrier media, in conjunction with Labroid as a surfactant. E.g. See US Pat. No. 4,388,307. The use of compositions similar to the drink solutions presented in this technique, however, involves various challenges.

Furthermore, the good taste of known oil based systems has proved problematic. The taste of known drink-solutions of some drugs is particularly unpleasant. Mixing with a moderately flavored drink is usually done immediately at high dilution prior to dosing, for example, to allow for normal treatment at a chocolate drink formulation, which is acceptable to all. The use of oil-based systems also requires the use of high ethanol concentrations, which are inherently undesirable by themselves, especially where pediatric administration is expected. Also, evaporation of ethanol is for example. From capsules (used in many parts, to solve the problem of good taste, as discussed or in other formulations (eg when opened)), progress is made in drug settling. When such a composition is presented, for example, in a soft gelatin encapsulation formulation, this particular difficulty requires the packaging of an airtight component, for example an airtight transparent pack or an aluminum foil blister pack, an encapsulated product. do. This in turn makes the goods bulkier, producing more expensive production. In addition, the storage characteristics of the aforementioned formulations are far from ideal yet.

The level of bioavailability obtained using existing oral administration systems for the macrocyclic immunosuppressive agents described herein is also low, and varying time between individuals during treatment, in individual patient types, and even in single individuals. Indicates a wide range of deviations. Reports in the literature show that current treatments using commercially available macromolecular immunosuppressant drink solutions provide only about 30% average absolute bioavailability, with marked deviations between populations. There is. For example, deviations between humans with liver (relatively low bioavailability) and bone marrow (relatively high bioavailability) transplants. Deviations in reported bioavailability among subjects vary from one or a few percent of some patients to 90% or more of other patients. As already stated, noticeable changes in individual bioavailability with time are frequently observed. Thus, there is a need for consistent and high bioavailability of many of the agents shown here in patients.

The use of such dosage forms is also characterized by extreme deviations in the dosage of the patient as needed. In order to carry out an effective treatment, the drug blood concentration or the serum serum concentration must be maintained within a certain range. This required range may in turn vary depending on the particular disease being treated, for example whether the therapy can prevent organ transplant rejection or whether it is for the control of an autoimmune disease or diseases thereof. And whether or not the alternative macromolecular immunosuppressant therapy will be used incidentally or not with any of the macromolecular immunosuppressants of the therapy of formula ad. Because of the wide variation in bioavailability obtained with existing dosage forms, the daily dosages necessary to obtain the required serum concentration will vary considerably between individuals and even for a single individual. For this reason, it will be necessary to monitor blood / serum concentrations at regular and frequent intervals in patients undergoing macrocyclic immunosuppressant treatment. Monitoring of blood / serum concentrations should be carried out on a regular basis. It is generally carried out using immunoassay techniques of RIA (radioimmunoassay) or equivalent effect, for example techniques based on monoclonal antibodies. This is inevitably time consuming and inconvenient and actually adds to the overall cost of the therapy.

In addition, the blood / serum concentrations of the macrocyclic immunosuppressants described herein obtained using applicable dosage systems may show extreme variations between peak and trough concentrations. It is for each patient and the effective macromolecular immunosuppressant concentration in the blood varies widely between each administration during individual dosing.

There is also a need to provide macrocyclic immunosuppressants in water-soluble formulations for injection dosing. It is well known that Cremaphore L (CreL), which is used in current formulations of macromolecular immunosuppressive agents, is a polyoxyethylated derivative of castor oil and a toxic transporter. There have been many instances of hypersensitivity due to the castor oil component. Currently, no formulations are available that allow these agents to reach the required concentrations in aqueous solutions because of the low solubility of macromolecular immunosuppressants in water.

Above all such very obvious practical difficulties, there is the occurrence of unwanted additional reactions already mentioned, which are observed using the applicable oral dosage forms.

Several proposals to address these various problems have been presented in the art, including oral dosage forms of both solidifying liquids. However, the primary difficulty still remaining is that macrocyclic immunosuppressants are inherently insoluble in aqueous media, which, by their nature, prevents the use of dosage forms that can contain macromolecular immunosuppressants in sufficiently high concentrations. Is a formulation that allows for convenient use but still achieves high absorption of blood / serum consistent with effective absorption from the lumen of the gastrointestinal or intestinal tract so as to meet the required criteria in terms of bioavailability. to be.

Certain difficulties encountered with oral administration of these macromolecular immunosuppressive agents inevitably limit the use of macromolecular immunosuppressive therapies to treat diseases of diseases that may be relatively less severe or dangerous. Is that. Part of the difficulty in this regard has been the introduction of macrocyclic molecular immunosuppressive therapies in the treatment of autoimmune diseases and other diseases affecting the skin, such as atopic dermatitis and psoriasis. As such, it is used for stimulating hair growth, for example, in the treatment of alopecia areata caused by old age or disease.

Thus oral macromolecular immunosuppressive therapies show that the medicament has significant potential benefits, for example, in patients suffering from psoriasis, while at the risk of side effects from oral therapy, preventing universal use. Various proposals have been made for the application of macrocyclic immunosuppressants, for example in this technique, in which cyclosporin and topical delivery systems in topical formulations have been described. However, several attempts at topical application have not provided significantly effective therapies.

However, the present invention overcomes the problems described above. More specifically, one embodiment of the present invention relates to a prodrug of a macrocyclic molecular immunosuppressive agent that significantly improves its solubility in aqueous solutions, whereby a carrier such as ethanol or castor oil is applied when administered in solution. There is no need to use it. In addition, the prodrugs of macrocyclic immunosuppressants do not exhibit the side effects of the formulations in this prior art, according to the present invention. Furthermore, the inventors have found that the macrocyclic immunosuppressive prodrugs of the present invention enhance oral uptake when administered to a patient in the form of a prodrug, thereby significantly improving its bioavailability and efficacy.

Thus, in one aspect, the invention relates to prodrugs of macrocyclic immunosuppressants. The prodrug includes an amino acid esterifying to one of the cyclohydroxysine, sirolimus, free hydroxyl groups present on the side chain of tacrolimus and hydroxyl groups of the pimecrolimus molecule.

For example, one aspect of the present invention relates to compounds of Formulas (a)-(d) or pharmaceutically acceptable salts thereof;

CYCLO in the formula shows residues at positions 2-11 of the cyclosporin molecule; -xy- is CH = CH or CH ₂ CH ₂ and AA is an amino acid or a dipeptide of the formula GLYAA. In the latter case, GLY is glycine and AA is any one amino acid. In the dipeptide structure, AA is attached to the agent via an OH group using glycine as a spacer. Glycine is esterified to cyclosporine, and then glycine is bound to AA by an amide bond using the amino group of glycine and the carboxylic acid group of AA.

The present invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of the above compounds of formula ad and a pharmaceutical carrier thereof.

In another embodiment, the present invention is directed to a method of treating a patient in need of a macrocyclic immunoreactive formulation therapy, the method comprising administering to a patient mentioned an effective amount of the compounds of formula ad.

In another embodiment, the invention provides a MeBmt at position 1 of a cyclosporin molecule as well as hydroxyl functionalities specified in formula bd having an acylating derivative of the agent under one amino acid or ester forming conditions. Macrocyclic molecular immunity in an aqueous solution comprising reacting with a hydroxy functional group of a moiety, or using a dipeptide structure that attaches a simple amino acid or AA to the agent using glycine as a spacer and separates the product of the drug. It relates to a method of improving the solubility of an inhibitor.

In another embodiment, the invention provides MeBmt at position 1 of the cyclosporin molecule as well as the hydroxyl functionalities specified in formula bd having an acylating derivative of the agent under amino acid or ester forming conditions. Reacting with the hydroxy functional group of the moiety, or attaching a simple amino acid or AA to the medicament using glycine as a spacer and isolating the product of the medicament, administering the mentioned product to the patient, and using a dipeptide structure. Including the present invention relates to a method for improving the bioavailability of macrocyclic immunosuppressive agents when administered to a patient.

summary:

The procedure for the synthesis of N- (L-proline) -glycine and N- (L-lysine) -glycine esters of cyclosporin A will be outlined in the Synthesis Sequence section. These schemes are illustrative examples and can be applied equally to other amino acids. The complete process and analytical data will be provided in the experimental section. Cyclosporin A (15 g) was combined with chloroacetic anhydride (4 equiv) in anhydrous pyridine. The experiment produced chloroacetate esters of cyclosporin A in good yield (SPI001201, 14 g, 88% yield). Chloroacetate ester (10.1 g) was then treated with sodium azide in DMF to produce azidoacetate ester of cyclosporin A (SPI001202, 9.9 g, 97% yield). Azidoacetate (9.8 g) was reduced to tin chloride (9 g) to prepare the glycine ester of cyclosporin A (8.54 g, 89% yield). The glycine ester of cyclosporin A (SPI001203) was then bound in an amount greater than twice that of either boc-L-proline or Boc-L-lysine using EDC as the binding agent. After purification by column chromatography, the boc protecting groups were removed from the dipeptide esters of cyclosporin A at low temperature 5 ° C. by treatment with 2M hydrochloric acid in diethyl ether. The L-glycine ester salt of cyclosporin A did not require any further purification and was then dried. The L-proline glycine ester salt of cyclosporin A requires a purification process. The salt was converted to free-base with sodium bicarbonate and purified through silica gel (eluting with acetone). The salt was then formed at low temperature with dilute anhydrous hydrochloric acid and then dried.

Synthetic sequence:

Synthesis of N- (L-proline) -glycine and N- (L-lysine) -glycine esters of cyclosporin A : a) pyridine; b) NaN ₃ , DMF; c) SnCl ₂ , methanol; d) boc-L-lysine, EDC; e) boc-L-proline, EDC; f) HCl, Et ₂ 0.

Part of the experiment:

SPI0022 and SPI0023 were performed in batches. In general, small scale experiments were performed initially and then as larger batches. Reagents mentioned in the experimental sections were purchased with the highest purity available from Aldrich, Acros or Bachem, with the exception of solvents purchased from either Fisher Scientific or Mallinkrodt. Cyclosporin A (USP grade) used in these procedures was provided by Signature Pharmaceuticals, Inc.

One) SPI001201

Cyclosporin A (15.01 g, 0.0124 moles) was dissolved in anhydrous pyridine (35 mL) at room temperature under argon atmosphere. The solution was cooled to 5 ° C. in an ice / water batch and chloroacetic anhydride (9.10 g, 0.053 moles) was added. After stirring for 10 minutes, the ice bath was removed and the solution was stirred for 17 hours at room temperature under argon atmosphere. After 17 hours, diethyl ether (200 mL) was added. The ether was washed with water (2x100 mL) and dried over sodium sulfate (10 g) for 1 hour. After filtration and concentration under reduced pressure, the remaining yellow foam was dried under high pressure vacuum conditions (1 hour at room temperature) and flash chromatography eluting with heptane / ethyl acetate (2: 1) on silica gel (200 g). Purification with After mixing and concentrating the product containing fractions, the remaining pale yellow foam (14.8 g) was finally purified through a crystallization reaction with heated diethyl ether (140 mL). After cooling (-10 ° C., 2 hours), filtration, and drying under high pressure vacuum, the process produced chloroacetate ester SPI001201 (14.0 g, 88.3% yield) of cyclosporin A as a white solid.

Cyclosporine A Chloroacetate Ester:

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 8.50 (d, 1H, J = 9.6 Hz), 7.95 (d, 1H, J = 6.6 Hz), 7.46 (d, 1H, J = 9.0 Hz), 7.40 (d, 1H, J = 7.8 Hz), 5.35-4.52 (m, 15H), 4.37 (t, 1H, J = 7.2 Hz), 4.12 (d, 1H, J = 14.7 Hz), 3.89 (d, 1H , J = 14.7 Hz), 3.45-3.0 (m, 15 H), 2.8-2.5 (m, 6H), 2.5-1.5 (m, 16H), 1.5-0.7 (m, 53 H).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 173.78, 173.37, 172.86, 172.61, 171.28, 171.18, 170.91, 170.79, 168.78, 167.64, 167.18, 128.77, 126.68, 75.46, 65.95, 58.89, 57.47, 55.31. , 54.86, 54.34, 50.19, 48.91, 48.35, 48.02, 44.80, 40.96, 39.44, 37.07, 35.93, 33.85, 33.25, 32.40, 31.74, 31.50, 30.38, 30.12, 29.82, 29.53, 25.13, 24.92, 24.78, 24.40, 24.40, 24.40 , 23.75, 22.85, 21.94, 21.41, 21.25, 20.84, 19.85, 18.79, 18.32, 17.89, 17.82, 15.46, 15.24, 10.08.

2) SPI001202

Chloroacetate ester SPI001201 (10.10 g, 7.89 mmole) of cyclosporin A was dissolved in anhydrous N, N-dimethylformamide (30 mL) at room temperature. Sodium azide (2.15 g, 33.0 mmole) was added. The mixture was stirred for 24 hours in the dark, at room temperature, under argon atmosphere. After 24 hours, diethyl ether (150 mL) was added and the precipitate was filtered off. The ether was washed with water (2x100 mL) and dried over sodium sulfate (15 g) for 30 minutes. After filtration and concentration under reduced pressure, the remaining white solid was dried at room temperature under high pressure vacuum for 1 hour. The experiment produced azidoacetate ester SPI001202 (9.90 g, 97% yield) of cyclosporin A as a white solid and was used without further purification.

Azidoacetate Ester of Cyclosporine A:

¹ H NMR (300 MHz, CDC13): δ = 8.48 (d, 1H, J = 9.3 Hz), 7.95 (d, 1H, J = 6.9 Hz), 7.45 (d, 1H, J = 9.0 Hz), 7.39 ( d, 1H, J = 7.8 Hz), 5.5-4.5 (m, 15 H), 4.31 (t, 1H, J = 6.6 Hz), 4.04 (d, 1H, J = 17.3 Hz), 3.53 (d, 1H, J = 17.3 Hz), 3.45-3.0 (m, 15 H), 2.8-2.5 (m, 6H), 2.5-1.5 (m, 16H), 1.5-0.7 (m, 53 H).

¹³ C NMR (75 MHz, CDCl ₃ ): δ = 173.76, 173.32, 172.82, 172.53, 171.13, 170.89, 170.76, 170.69, 169.70, 168.20, 167.49, 128.63, 126.61, 74.96, 58.91, 57.39, 55.56,55.21, 54.80 , 54.23, 50.14, 48.99, 48.23, 48.24, 47.93, 44.71, 40.89, 39.33, 39.22, 37.02, 35.83, 33.81, 32.96, 32.31, 31.67, 31.42, 30.31, 30.09, 29.76, 29.47, 25.08, 24.92, 24.84, 24.84, 24.84 , 24.51, 24.40, 23.94, 23.82, 23.71, 21.85, 21.33, 21.25, 20.82, 19.79, 18.71, 18.25, 17.92, 17.81, 15.17, 10.03.

3) SPI001203

The azidoacetate ester of cyclosporin A SPI001202 (9.80 g, 7.62 mmole) was dissolved in methanol (250 mL) at room temperature. Water (40 mL) was added followed by tin (II) chloride (5 g, 26.3 mmole). When tin (II) chloride (4 g, 21.0 mmole) was added in an additional amount, the solution was allowed to stir at room temperature for 1 hour. The solution was allowed to stir additionally for 2 hours at room temperature. Water (200 ml) containing ammonium hydroxide (40 mL, 29%) was added. After filtration, the solution was concentrated under reduced pressure (until 200 mL). The remaining aqueous solution was extracted with ethyl acetate (2x200 mL). Ethyl acetate fractions were combined, dried over sodium sulfate (20 g), filtered under reduced pressure and concentrated. The remaining clear foam was purified by filtration over silica gel (150 g), eluting with dichloromethane / methanol (20: 1). The process produced glycine ester of cyclosporin A (8.54 g, 89% yield) in a clear, solid foam.

Glycine esters of cyclosporin A:

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 8.60 (d, 1H, J = 9.6 Hz), 8.06 (d, 1H, J = 6.9 Hz), 7.53 (d, 1H, J = 8.4 Hz), 7.51 (d, 1H, J = 6.6 Hz), 5.7-4.52 (m, 15 H), 4.41 (t, 1H, J = 6.9 Hz), 3.5-3.0 (m, 17 H), 2.82-2.5 (m, 8H ), 2.5-1.5 (m, 16H), 1.5-0.7 (m, 53H).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 174.10, 173.67, 173.23, 172.72, 172.55, 171.18, 171.10, 170.73, 170.61, 169.68, 167.77, 128.82, 126.42, 73.83, 58.57, 57.32, 55.99, 55.20, 54.54, 54.54 , 54.31, 50.08, 48.82, 48.28, 47.90, 44.70, 43.81, 40.74, 39.33, 39.24, 37.02, 35.84, 33.72, 33.07, 32.39, 31.72, 31.41, 30.25, 29.98, 29.74, 29.51, 25.05, 24.81, 24.73, 24.73, 24.73 , 24.31, 23.91, 23.78, 23.68, 21.86, 21.33, 21.25, 20.68, 19.76, 18.74, 18.24, 17.94, 17.79, 15.18, 10.03.

4) SPI0022

Glycine esters of cyclosporin A (SPI001203, 2.0 g, 1.59 mmole) were combined with Boc-L-lysine (1.31 g, 3.78 mmole) and EDC (0.75 g, 3.9 mmole) at room temperature under argon atmosphere at room temperature with dichloromethane (25 mL). Boc-L-lysine was prepared from dicyclohexylamine salt (2.0 g in 50 mL ether) by extraction with cold potassium hydrogen sulfate solution (1 g in 50 mL water) and then Extracted with chilled water (2x50 mL). The ether containing Boc-L-lysine was dried over sodium sulfate (5 g), filtered, concentrated under reduced pressure, and then dried under high pressure vacuum at room temperature for 1 hour. Several crystals of DMAP were added to the mixture of EDC, Boc-L-lysine, and the glycine ester of cyclosporin A and the solution was allowed to stir at room temperature for 4 hours. Dichloromethane solution was extracted with DIUF water (50 ml), 5% sodium bicarbonate solution (50 ml), and DIUF water (50 ml). After drying over sodium sulfate (10 g), the dichloromethane solution was filtered and concentrated under reduced pressure. The remaining white foam (3.01 g) was purified by flash chromatography using silica gel (50 g) eluting with heptane / acetone (2: 1). The product containing fractions were combined and concentrated under reduced pressure and then dried under high pressure vacuum conditions. The purified and protected intermediate (2.34 g, white solid, 92.8% yield) was left into the flask under argon atmosphere and cooled in an ice water bath. Cold anhydrous 2M hydrochloric acid was added to diethyl ether (20 ml) and the solution was then stirred at 5 ° C. for 8 hours. The mixture was allowed to slowly warm up to room temperature overnight. After stirring for a total of 20 hours, the flask was cooled again in an ice water bath for 30 minutes. The product was filtered and dried at room temperature, high pressure vacuum conditions for 1 hour and then at 50 ° C. for 4 hours. The experiment produced N- (L-lysine) -glycine ester, dihydrochloride trihydrat (SPI0022,1.59 g, 73.9% yield) of cyclosporin A as a white solid.

¹ H NMR (300 MHz, CDCl ₃ , NMR data is free base): δ = 8.58 (d, 1H, J = 9.3 Hz), 8.04 (d, 1H, J = 6 Hz), 7.80 ( d, 1H, J = 6 Hz), 7.49 (d, 2H, J = 8.4 Hz), 5.70-4.6 (m, 17H), 4.41 (m, 1H), 4.28 (dd, 1H, J = 17, 7.2 Hz ), 3.67 (d, 1H, J = 17 Hz), 3.46 (s 3H), 3.4-2.8 (m, 16 H), 2.8-2.5 (m, 8H), 2.5-1.35 (m, 24H), 1.5- 0.7 (m, 50 H).

¹³ C NMR (75 MHz, CDCl ₃ , NMR data is free base): δ = 175.23, 173.77, 173.34, 172.75, 172.63, 171.34, 171.22, 170.94, 170.84, 170.91, 169.89, 169.70, 128.74, 126.67, 74.41, 58.82, 57.43, 55.91, 55.21, 54.81, 54.42, 50.17, 48.89, 48.31, 47.98, 44.78, 41.92, 40.82, 40.69, 39.44, 39.32, 27.19, 35.91, 34.88, 33.71, 33.25, 33.12, 33.12, 32.12 31.83, 31.50, 30.38, 30.06, 29.81, 29.55, 25.14, 24.90, 24.52, 24.43, 24.00, 23.76, 21.93, 21.42, 21.29, 20.81, 19.84, 18.82, 18.32, 17.96, 17.86, 15.21, 10.10.

CHN analysis:

Calculated for C ₇₀ H ₁₂₈ Cl ₂ N ₁₄ O ₁₅ -3H ₂ O: C 55.50, H 8.92 and N 12.74; found: C 58.28, H 8.98 and N 13.16.

HPLC analysis:

99.60% purity; rt = 14,763 min .; 80% acetonitrile / 20% Tris base in DIUF water; 1 mL / min; 60 C; Synergi Hydro RP, 4u column (serial # 163383-7), 4.6 × 250 mm; 20 ul; UV = 210 nm.

Melting Point: 196.0-198 ℃ (Uncalibrated)

5) SPI0023

Glycine esters of cyclosporin A (SPI001203, 7.50 g, 5.95 mmole) were combined with Boc-L-lysine (2.56 g, 11.90 mmole) and EDC (2.28 g, 11.9 mmole) anhydrous dichloromethane (50) at room temperature under argon atmosphere. mL). Several crystals of DMAP were added to the mixture of EDC, Boc-L-lysine, and the glycine ester of cyclosporin A and the solution was allowed to stir at room temperature for 3 hours. The dichloromethane solution was extracted with DIUF water (50 ml), 5% sodium bicarbonate solution (2x50 mL), and DIUF water (50 ml). After drying over sodium sulfate (10 g), the dichloromethane solution was filtered and concentrated under reduced pressure. The remaining white foam (9.50 g) was purified by flash chromatography using silica gel (150 g), eluting with heptane / acetone (2: 1 followed by 1: 1). The product containing fractions were combined and concentrated under reduced pressure, then dried at room temperature under high pressure vacuum for 10 minutes (7.94 g white solid, 91.7% yield). The purified and protected intermediate (6.46 g) was left into the flask under argon atmosphere and cooled in an ice water bath. Cold anhydrous 2M hydrochloric acid was added to diethyl ether (150 ml) and the solution was then stirred at 5 ° C. for 8 hours. The mixture was allowed to slowly warm up to room temperature overnight. After stirring for a total of 20 hours, the flask was cooled again in an ice water bath for 30 minutes. The product was filtered off and dried under high pressure vacuum conditions at room temperature for 30 minutes. N- (L-proline) -glycine ester of cyclosporin A, hydrochloride (5.17 g, 84.6% yield, and 90% purity using HPLC) was DIUF water (25 ml) containing sodium bicarbonate (1 g) It was converted to the free base by dissolving the salt in. The free base was extracted with dichloromethane (3x25 mL), after drying over sodium sulfate (5 g), filtered and concentrated. The remaining yellowish white solid (5 g) was purified by filtration through silica gel (100 g), eluting with acetone. The product containing fractions were combined and concentrated under reduced pressure, then dried at high pressure vacuum for 30 minutes at room temperature. The hydrochloride was regenerated by dissolving the free base (3.8 g) in diethyl ether (25 mL) and adding the solution 2M hydrochloric acid (5 mL) in heptane (50 mL) to the solution and cooling in an ice water bath. After 20 minutes at 5 ° C., the white solid was filtered and dried under high pressure vacuum conditions at room temperature for 6 hours. The experiment produced N- (L-proline) -glycine ester of cyclosporin A, hydrochloride (SPI0023, 3.8 g) as a white solid.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 14.20 (br s, 2H), 8.62 (d, 1H, J = 10 Hz), 8.06 (d, 1H, J = 6.9 Hz), 7.61 (d, 1H, J = 8.1 Hz), 7.48 (d, 1H, J = 9 Hz), 5.70-5.50 (m, 3H), 5.40-4.60 (m, 12H), 4.37 (m, 1H), 4.20 (d, 1H , J = 18 Hz), 3.97 (d, 1H, J = 18 Hz), 3.70 (m, 1H), 3.45 (s, 3H), 3.23-3.08 (m, 12H), 2.66 (s, 3H), 2.60 (s, 3H), 2.50-1.80 (m, 15H), 1.78-1.20 (m, 15H), 1.15-0.66 (m, 46H).

¹³ C NMR (75 MHz, CDCl ₃ ): a = 174.15, 173.49, 172.67, 172.59, 171.86, 171.20, 171.13, 171.02, 170.83, 169.68, 168.77, 167.55, 128.30, 127.10, 80.09, 75.58, 62.65, 59.35. , 55.53, 55.30, 54.78, 54.35, 53.60, 50.25, 50.09, 48.92, 48.18, 48.12, 44.62, 40.59, 40.02, 39.43, 39.30, 37.13, 35.88, 33.74, 33.07, 32.19, 32.01, 31.86, 31.50, 31.43, 43.43 , 29.93, 29.72, 29.30, 29.16, 27.56, 26.04, 25.00, 24.86, 24.74, 24.39, 20.96, 19.81, 18.71, 18.26, 18.09, 17.85, 17.79, 15.09, 14.30, 10.00.

CHN analysis: calculated for C ₆₉ H ₁₂₂ ClN ₁₃ 0 ₁₄ : C 59.48, H 8.83 and N 13.07; found: C 59.84, H 9.02 and N 12.65.

HPLC analysis: 99.59% purity; rt = 10.613 min .; 85% acetonitrile / 15% Tri base in DIUF water; 1.2 mL / min; 60C; Synergi Hydro RP, 4u column (serial # 163383-7), 4.6 × 250 mm; 20 ul; UV = 210 nm.

Melting Point: 197.0-199 ℃ (Uncalibrated)

Prodrugs of macrocyclic immunosuppressants of the present invention are effective in the treatment of diseases or disorders in which macrocyclic immunosuppressants are normally used. The prodrugs disclosed herein enhance the therapeutic advantages of macrocyclic immunosuppressants by reducing or eliminating the biopharmaceutical and pharmacokinetic barriers that are modified in the human body to release the active compound and are associated with each of the prodrugs. Let's do it. However, it should be noted that these prodrugs themselves will have sufficient activity in mammals without releasing any active agent. Since the prodrugs are more soluble in water than cyclosporine or other macromolecular immunosuppressants, they should be associated with carrier carriers such as alcohol or castor oil, which may be toxic or cause unwanted additional reactions. no need. In addition, oral formulations containing prodrugs of macrocyclic immunosuppressants are absorbed into the blood and are quite effective.

Thus, the prodrugs of the cyclosporine of the present invention enhance the therapeutic advantages by removing the biopharmaceutical and pharmacokinetic barriers of existing agents.

Furthermore, the prodrugs are easily synthesized in high yields easily and using currently available materials.

VI . Valproic acid ( Valproic Acid ) Esters

Valproic acid (2-Propylpentanoic acid) is a drug widely used as an anticonvulsant drug. It is a low molecular weight carboxylic acid derivative useful for epilepsy and is also used for brain mitigation. Vasodilator effect in The drug is administered orally to control epilepsy in humans and also relieves severe pain associated with migraine headaches.

Valproic acid has been shown to have a very large number of therapeutic applications, which is quite variable and somewhat surprising, for example, in addition to its efficacy in the treatment of epilepsy and migraine headaches, which is effective in the treatment of any psychiatric disorders. Such as bipolar disorder, emotional stabilization, aggressive control, impulsiveness in personality disorders, fluctuations in dementia, and in the treatment of post traumatic stress disorder (PTSD). It has been used as adjuvant therapy.

Mechanism of action:

Although it has been used for the treatment of epilepsy for many years, the exact mechanism of action of valproic acid is still unknown. Valproic acid has been claimed to exert its action by increasing the concentration of gamma-amino butyric acid (GABA) in the brain. Gamma-amino butyric acid is a neurotransmitter, a chemical used by nerves to communicate with other nerves.

Valproic acid is used for myoclonic epilepsy, with or without generalized tonic-clonic seizures, including pediatric myoclonic epilepsy of Janz, beginning in adolescence or early adulthood. It is the drug of choice in treatment. Photosensitive myoclonus is usually easily controlled. Valproate is also effective in the treatment of benign myoclonic epilepsy and postanoxic myoclonus, and, along with clonazepam, by strain-tolerant seizures. It is also effective against characterized progressive developmental myoclonic epilepsy. It would also be desirable in any stimulus-sensitive (reflection, surprise) epilepsy.

Valproate may be effective in infantile spasms; Convulsions are relatively contradictory in children who have been caused by hyperglycemia or other metabolic (mitochondrial) abnormalities. In general, it is difficult to control atonic and akinetic seizures in patients with Lennox-Gastaut syndrome, but Valproate can be selected for treatment of mixed seizure types. It is a drug. Since this drug has been useful in some patients resistant to all other antiepileptic drugs, it may warrant trials in almost all unresponsive patients regardless of the type of seizure.

Despite the usefulness of Valproate, hepatotoxicity can be fatal, but it is specific and can be prevented by routinely monitoring liver enzymes. Hepatotoxicity occurs in very young children and is most common in humans receiving multiple anticonvulsants. Valproate-induced cytopenias will be dose-related and will ensure monitoring of complete blood cell counts during treatment.

Encephalopathy with hyperammonemia can occur without abnormal liver function tests. Women in their first months of pregnancy are at risk for neural tube defects.

Valproic acid is a low molecular weight liquid with a characteristic odor. When taken orally, the drug has an unpleasant taste and can be very irritating to the mouth and throat. In order to convert Valproic acid into a solid dosage form convenient for oral administration, many derivatives with covalent and ionic bonds with carboxylic acids have been made. The simple sodium salt of valproic acid becomes the valproate sodium available as a solid. However, a stable coordination complex known as Divalproex sodium was formed by partial neutralization of two molecules of valproic acid with one atom of sodium. This product is the most widely available Valproic acid hemi salt marketed by Abbott Laboratories in the United States under the brand name Depakote®. Depakote® is also available in delayed release formulations for oral administration.

A significant disadvantage of Valproic acid is that it is difficult to administer in pharmaceutical formulations. Furthermore, administration of Valproic acid to other formulations may not produce consistently desired bioavailability. For example, the overall bioavailability of delayed release formulations of Valproate, its sodium salt, Divalproex®, and their agents from Valproic acid is not quite complementary. Since it is essential to continuously monitor the plasma profile of valproic acid, any change in plasma concentrations, due to changes in the formulation, may have a counterproductive effect on the overall treatment outcome.

In order to improve the therapeutic effect, to make the blood profile consistent, to develop pharmaceutically ordered formulations, and to reduce first-pass metabolism, the present invention overcomes some of the difficulties described above with prodrugs of valproic acid. Discuss them.

To date, there have been no commercially available pharmaceutical formulations that can deliver Valproic acid without harmful side effects. The present invention, however, has produced a number of water-soluble and non-toxic derivatives of valproic acid that are suitable for delivering valproic acid continuously in the human body without the need for harmful side effects, without the need for expensive additives, and without the need for excipients.

Thus, the present invention relates to one class of prodrugs of Valproic acid. The prodrug includes hydroxyl groups of amino acids esterified with free carboxyl groups present on the Valproic acid molecules. In another embodiment, the amine group of the amino acid reacts with the COOH group to form an amide bond.

More specifically, one embodiment of the present invention relates to compounds of the formula below or pharmaceutically acceptable salts thereof;

Wherein R in the formula is either NH-AA or O-AA, AA is an amino acid, and either an amine group or hydroxyl group reacts with the carboxylic acid group of valproic acid.

The invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a variety of the above Valproic acid prodrugs and pharmaceutical carriers thereof.

In another embodiment, the invention relates to a method of treating a patient in need of Valproic acid therapy, the method comprising administering to the patient an effective amount of Valproic acid. .

In another embodiment, the present invention relates to a method for converting liquid valproic acid into a solid powder by reacting one of the amino or hydroxyl functional groups of an amino acid with a carboxyl functional group of valproic acid and separating their products. .

In another embodiment, the present invention is directed to a method that is a practically and therapeutically effective means. To reduce or eliminate the potential first-pass metabolism and thereby improve the sustained therapeutic effect by administering the prodrug to the patient, the prodrug being Valproic acid to form an ester or amide covalent bond, respectively Carboxylic acid functional groups of the molecule include reacting an amine functional group or a hydroxyl functional group of selected amino acids and isolating their products to administer the product to the patient.

When naturally occurring unsubstituted amino acids are esterified to Valproic acid, the resulting prodrugs are pharmaceutically ordered free flowing powders, rapidly absorbed into the body, and non-toxic as they split in the body. It has been found that it releases amino acids and requires none of the toxic emulsifiers, additives and other excipients.

Furthermore, the present invention also produced drugs, which are prodrugs of valproic acid of the present invention; The drugs are very effective antiepileptic drugs and have shown to remain so intact. Current amino acid prodrugs are therefore effective anti-epileptic drugs and are useful in the treatment of numerous psychiatric disorders and exhibit such potential without releasing an effective parent drug.

The bulk density of the Valproic acid prodrug is much higher than the corresponding sodium salts, and the agents are suitable for compressing large weight tablets and capsules. Furthermore, they do not have a bitter taste or unusual smell of Valproic acid.

The prodrugs of the present invention do not have to have such action due to the blocking of the carboxylic acid groups responsible for possessing acidic action, while the prodrugs are effective anti-epileptic drugs with or without the release of valproic acid. It is being seen. However, all the Valproic acid prodrugs described above are released as in vivo experiments as effective agents with the pharmacological and psychoactive properties of all of the agents.

Valproic acid prodrugs obviously have many advantages of valproic acid, such as all the side chains cleaved from such prodrugs are naturally occurring essential amino acids and thereby non-toxic. This leads to a high therapeutic index. Secondly, all of its prodrugs are easily broken into the body to release valproic acid. Furthermore, because of the high water solubility of the prodrugs they can be easily administered, forming an in - situ solution just prior to administration by intravenous injection with lyophilized sterile powder, or for infusion. Administration may be by providing a solution in prefilled syringes or bottles. Amino acid esters are more stable than Valproic acid because the COOH group in Valproic acid is blocking the reaction with bases. Thus, the prodrugs of Valproic acid of the present invention are more effective than Valproic acid itself, which is non-toxic and without the other pharmaceutical problems associated with currently marketed formulations.

The procedure for the synthesis of L-serine, L-threonine, and L-hydroxyproline esters of valproic acid (2-propylpentanoic acid) will be outlined in the Synthesis Sequence section. These schemes are illustrative examples and can be applied equally to other amino acids. The complete process and analytical data will be provided in the experimental section. In general, Valproic acid (2-8 g in batches) was combined with N-benzyloxy / benzyl ester protecting amino acids using EDC in the presence of a catalytic capacity of DMAP. After the reactions were complete (20 hours at room temperature), the mixture was extracted with DIUF water, dried over sodium sulfate and concentrated under reduced pressure. The raw material was immediately used for the deprotection step or otherwise purified by column chromatography. The process produced protected esters of valproic acid in yields ranging from 72% to 92%. The protecting groups were removed by hydrogenation (30 psi hydrogen) in the presence of 10% palladium on carbon. The amino acid esters of valproic acid were extracted from the palladium catalyst with ethanol, concentrated and then dried. The final salt was formed by acidification of hydrochloric acid. The raw salts (from 57% to 92% yield) were then purified by the methods described in the experimental section.

Synthetic sequence:

One. SPIC001

2. SPIC002

3. SPIC003

Synthesis of L-serine, L-threonine, and L-hydroxyproline esters of Valproic acid: a) EDC, DMAP, CH ₂ Cl ₂ ; b) H ₂ , 10% Pd / C, EtOH, EtOAc; c) HC1.

Part of the experiment:

The synthesis of SPICOOI, SPIC002 and SPIC003 was performed in batches. In general, small scale experiments were performed initially and then as larger batches. The materials mentioned in the experimental section were purchased with the highest purity available from Lancaster, Sigma-Aldrich, Acros, or Bachem, with the exception of solvents purchased from either Fisher Scientific or Mallinkrodt.

1) SPIC001 : 2-propylpentanoic acid 2 (S) -amino-2-carboxy-ethyl ester, hydrochloride (L- serine- valproic acid ester, hydrochloride)

2-propylpentanoic acid (valproic acid, 6.48 g, 44.93 mmole) in anhydrous dichloromethane (50 mL), N-carbenzyloxy-L-serine benzyl ester (Z-Ser-OBzl, 14.80 g, 44.93 mmole), EDC (8.61 g, 44.91 mmole) and DMAP (549 mg, 4.49 mmole) were stirred at room temperature under argon atmosphere for 20 hours. After 20 hours, the dichloromethane layer was washed again with water (3x50 mL), dried over magnesium sulfate (5 g), then under reduced pressure, filtered and concentrated. The remaining colorless oil (20.87 g) was purified by flash chromatography on silica gel (150 g, 0.035-0.070 mm, 6 nm pore diameter), eluting with hexane / ethyl acetate (3: 1). After concentration of the product containing fractions under reduced pressure and drying under high pressure vacuum until its weight became constant, the experiment produced L-serine ester of Valproate as colorless oil (18.9 g, 92% yield).

¹ H NMR (300 MHz, DMSO): δ = 7.96 (1H, d, J = 8. 1 Hz), 7.35 (1OH, m), 5.14 (2H, s), 5.05 (2H, s), 4.51 (1H , m), 4.29 (2H, m), 2.29 (1H, m), 1.50-1. 25 (4H, m), 1. 25-1. 10 (4H, m), 0.80 (6H, t, J = 6.6 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 174.88, 169.15, 155.85, 136.58, 135.45, 128.26, 128.18, 127.47, 127.71, 127.57, 66.32, 65.66, 62.47, 53.09, 44.20, 33.86, 33.79, 19.95, 13.85.

Protected L-serine-valproate ester SPIC00101 (18.9 g, 41.48 mmole) was dissolved in ethanol (60 mL) and ethyl acetate (60 mL) at room temperature, and under nitrogen atmosphere, 10% palladium (3.0 g, Poured into a Parr jar (500 mL) containing 50% wet). Nitrogen atmosphere was replaced with hydrogen gas (30 psi). After stirring for 4 hours, additional palladium catalyst (1.0 g) was added to ethanol and ethyl acetate (1: 1,100 mL) and the reaction mixture was stirred overnight at room temperature in hydrogen gas (30 psi). The ethanol and water fractions were concentrated under reduced pressure at room temperature. After drying under high vacuum, the remaining white solids were acidified with hydrochloric acid in diethyl ether (2M, 24.6 mL). The mixture was stored in the refrigerator before filtration and washing with additional cold diethyl ether (10 mL). After the filtration process, the remaining solids were dried (24 hours) at room temperature until constant weight was obtained under high pressure vacuum conditions. This experiment produced L-serine ester of Valproic acid, hydrochloride SPICOO1 (6.34 g, 57% yield) as a white solid,

^I H NMR (300 MHz, DMSO): δ = 8. 73 (br s, 3H), 4.47 (dd, 1H, J = 12. 9, 4.5 Hz), 4.31 (dd, 2H, J = 12. 9,3. 6 Hz), 2.36 (m, 1H), 1.50 (m, 2H), 1.39 (m, 2H), 1. 20 (m, 4H), 0.84 (t, 6H, J = 7 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 174.67, 168.19, 61.84, 51.16, 44.12, 33.76, 33.58, 20.07, 19.92, 13.97, 13.89.

HPLC analysis: 98.49% purity; rt = 4.767 min; Luna C18 5u column (sn 167917-13); 4.6 x 250 mm; 254 nm; 33% ACN / 66% DIUF number; 35 C; 20 ul inj. 1 ml / min; 20 mg / mL sample size; Sample dissolved in mobile phase.

CHN analysis: calc .: C 49.34, H 8.28, N 5.23; found: C 49.22, H 8.35, N 5.24.

Melting point: 159-160 ℃

2) SPIC002 : 4 (R)-(2-)-propyl-pentanoyloxy) -pyrrolidine-2 (S) -carboxylic acid (L-hydroxyproline-valproic acid ester)

2-propylpentanoic acid (valproic acid, 4.32 g, 30 mmole), N-carbenzyloxy-L-hydroxyproline benzyl ester (Z-Hyp-OBzl, 10.66 g, 30 mmole) in anhydrous dichloromethane (30 ml) ), ¹ , EDC (5.74 g, 30 mmole), and a mixture of DMAP (366 mg, 3 mmole) were stirred at room temperature under argon atmosphere for 20 hours. After 20 hours, the dichloromethane layer was washed again with water (3x30 mL) and dried over magnesium sulfate (5 g), then under reduced pressure, filtered and concentrated. The remaining colorless oil was used without purification. SPIC00201 (11.95 g, 24.7 mmole, 82.4% yield).

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.29 (1OH, m), 5.28-5.00 (5H, m), 4.55 (1 / 2H, t, J = 8 Hz), 4.46 (1 / 2H, t , J = 8 Hz), 3.80-3.60 (2H, m), 2.43-2.16 (3H, m), 1.60-1.45 (2H, m), 1.40-1.32 (2H, m), 1.28-1.20 (4H, m ), 0. 86 (6H, m).

¹³ C NMR (75 MHz, DMSO): δ = 174.74, 171.40, 171.05, 153.79, 153.31, 136.34, 136.20, 135.57, 135.38, 128.24, 128.13, 127.95, 127.87, 127.67, 127.52, 127.28, 127.10, 72.29 66.34, 66.10, 57.66, 57.19, 52.27, 51.89, 44.13, 40.33, 35.78, 34.79, 34.04, 33.92, 33.35, 20.00, 19.91, 13.79, 13.73.

Protected L-hydroxyproline-valproate ester SPIC00201 (17.24 g, 35.79 mmole) was dissolved in ethanol (50 mL) and ethyl acetate (100 mL) at room temperature and under nitrogen atmosphere, 10% palladium (3.5) g, 50% wet), was poured into a Parr bottle (500 mL). Nitrogen atmosphere was replaced with hydrogen gas (30 psi). After stirring for 15 hours, the catalyst was removed by a thin layer of celite and filtration through activated carbon. The ethanol and ethyl acetate mixture was concentrated under reduced pressure at room temperature. After drying under high pressure vacuum at room temperature overnight, the experiment yielded L-hydroxyproline-valproic acid ester SPIC002 (9.2 g, 99.8% yield) as a white solid. To remove trace impurities, amphoteric ions were purified using reverse-phase column chromatography (50 g ODS silica gel) in two batches. The zwitterion was placed in a column in DIUF water and eluted with DIUF water / methanol (2: 1, 1: 1, 1: 2, 100% methanol). The product containing fractions were mixed and concentrated at 20 ° C. (or below) under reduced pressure, then dried under high pressure vacuum at room temperature until constant weight was obtained (24 h, 6.4 g, white). Solid recovery).

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 12.40 (br s, 1H), 8.32 (br s, 1H), 5.28 (m, 1H), 4.11 (t, 1H, J = 7.2 Hz), 3.59 ( m, 1H), 3.34 (br d, 1H, J = 0.5 Hz), 2.50-2.22 (m, 3H), 1.62-1.50 (m, 2H), 1.50-1.32 (m, 2H), 1.32-1.19 (m, 4H), 0.88 (t, 6H, J = 7.2 Hz).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 175.99, 173.35, 71.83, 59.56, 49.77, 45.08, 36.19, 34.51, 20.87, 14.31.

HPLC analysis: 99.20% purity; rt = 7.228 min .; 70% DIUF water / 30% acetonitrile; 1 mL / min; 36.8C; Luna C18, 5u column (serial # 167917-13), 4.6x250 mm; 22 ul injection; Sample dissolved in mobile phase.

CHN analysis: calc .: C 60.68, H 9.01, N 5.44; found: C 60.58, H 9.12, N 5.48.

Melting Point: 179.0-180.0 ℃

3) SPIC003: 2-propyl-pentanoic acid 2 (S) -amino-2-carboxy-l (R) -methyl-ethyl ester, hydrochloride

(L-Threonine-Valproic Acid Ester, Hydrochloride)

2-propylpentanoic acid (valproic acid, 4.32 g, 30 mmole), N-carbobenzyloxy-L-threonine benzyl ester (Z-Thr-OBzl, 10.30 g, 30 mmole) in anhydrous dichloromethane (30 mL) , A mixture of EDC (5.74 g, 30 mmole), and DMAP (366 mg, 3.0 mmole) was stirred at room temperature under argon atmosphere for 20 hours. After 20 hours, the dichloromethane layer was washed again with water (3x30 mL) and dried over magnesium sulfate (5 g), then under reduced pressure, filtered and concentrated. The remaining colorless oil (13.44 g) was purified by column chromatography on silica gel (100 g, 0.035-0.070 mm, 6 nm pore diameter), eluting with hexane / ethyl acetate (4: 1). After concentration of the product containing fractions under reduced pressure and drying under high pressure vacuum conditions until the weight became constant, the experiment yielded protected L-threonine-valproate ester SPIC00301 (12.65 g, 89.8% yield). Created as a colorless oil

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.40-7.05 (11H, m), 5.45 (1H, m), 5.17-5.02 (4H, m), 4.53 (1H, d, J = 9.6 Hz), 2.24 (1H, m), 1.58-1.40 (2H, m), 1.40-1.15 (9H, m), 0.86 (6H, m).

¹³ C NMR (75 MHz, DMSO): δ = 174.24, 169.29, 156.48, 136.61, 135.34, 128.26, 128.20, 127.74, 127.67, 127.58, 69.04, 66.33, 65.78, 57.62, 44.50, 33.89, 33.80, 20.03, 19.91 16.40, 13.87.

Protected L-threonine-valproate ester SPIC00301 (12.65 g, 26.9 mmole) was dissolved in ethanol (50 mL) and ethyl acetate (50 mL) at room temperature, and under nitrogen atmosphere, 10% palladium (2.53 g, Poured into a Parr jar (500 mL) containing 50% wet). Nitrogen atmosphere was replaced with hydrogen gas (30 psi). After 20 hours, the catalyst was removed by filtration through a thin layer of activated carbon and washed with ethanol (25 mL). The ethanol and ethyl acetate mixture was concentrated under reduced pressure at room temperature. After drying under high vacuum, the remaining white solids (6.13 g) were acidified with hydrochloric acid (3.1 mL conc.) In DIUF water (50 mL). The solution was filtered through a second thin layer of activated carbon and dried overnight in a freeze-dryer. This experiment produced L-threonine-valproic acid ester, hydrochloride SPIC003 (6.52 g, 86.0% yield) as a white solid,

Mixed batches of L-threonine-valproic acid ester, hydrochloride SPIC003 (8.8 g) were purified by crystallization reaction from acetonitrile. After dissolving in heated acetonitrile (225 mL), the material was treated with activated carbon, filtered and then left at 5 ° C. in a refrigerator overnight. The white solids were filtered after 18 hours, washed with cold acetonitrile (10 mL) and then dried under high pressure vacuum conditions until the weight became constant (24 hours). The procedure recovered L-threonine-valproic acid ester, hydrochloride SPIC003 (6.82 g, 77.5% recovery) as a white solid.

¹ H NMR (300 MHz, DMSO): δ = 8.71 (br s, 3H), 5.28 (m, 1H), 4.16 (d, 1H, J = 2.7 Hz), 2.33 (m, 1H), 1.56-1.40 ( m, 2H), 1.37-1.27 (m, 5H), 1.21-1.13 (m, 4H), 0.84 (t, 6H, J = 6.6 Hz).

¹³ C NMR (75 MHz, DMSO): δ = 173. 97,168. 19,67. 69,55. 42,44. 43,33. 95,33. 78, 20.07, 19.95, 16.54, 13.94.

HPLC analysis: 98.88% purity; rt = 4.864 min .; 70% DIUF water / 30% acetonitrile; 1 mL / min; 40C; Luna C18, 5u column (serial # 211739-42), 4.6 × 250 mm; 20 ul injection; Sample dissolved in mobile phase.

CHN analysis: calc .: C 51.15, H 8.59, N 4.97; found: C 51.29, H 8.59, N 4.98.

Melting Point: 144 ℃

The solubility of the esters was determined in water at room temperature by dissolving excess of each agent to settle for several hours. The resulting solutions were centrifuged at 1500 rpm for 3 minutes and the supernatant was analyzed. These esters were shown to have solubility in water in excess of 50 mg / mL.

There are many screening experiments for determining the use of the prodrugs invented in accordance with the disclosed methods. These experiments include screening methods for both in vitro and in vivo experiments.

In vitro methods include acid / base hydrolysis of prodrugs, hydrolysis in porcine pancreas, hydrolysis in rat intestinal fluid, hydrolysis in human gastric juice, hydrolysis in human intestinal fluid, and hydrolysis in human plasma. These tests are described in Simmons, DM, Chandran, VR and Portmann, GA, Danazol amino Acid Prodrugs: In Vitro and In Situ Biopharmaceutical Evaluation, Drug Development and Industrial Pharmacy, Vol 21, Issue 6, Page 687,1995, all of which are incorporated by reference.

The compounds of the present invention are effective in treating diseases or disorders in which prodrugs of valproic acid are normally used. The prodrugs disclosed herein enhance the therapeutic benefits of valproic acid by reducing or eliminating the biopharmaceutical and pharmacokinetic barriers that are modified in the human body and associated with their individual to release the active compound. However, it should be noted that these prodrugs themselves will have sufficient functionality without releasing any effective agent in mammals.

Thus, the prodrug of the present invention enhances its therapeutic advantages by eliminating the biopharmaceutical and pharmacokinetic barriers of existing agents.

Furthermore, the prodrugs are easily synthesized in high yields, using materials that are easy to use and are currently available.

VII Fibric acid ( FIBRIC ACID Water solubility of derivatives Prodrugs

Fibric acid derivatives are useful for the treatment of hyperlipidemia in mammals whose symptoms are elevated triglycerides, low HD (high density lipoproteins or "good" cholesterol), and elevated cholesterol. Hyperlipidemic drugs. Fibric acid derivatives are also useful for reducing LDL (Low density lipoproteins, or "bad" cholesterol). The general structure of fibric acid analogs is shown below, where X is a variety of mixed aliphatic and aromatic functional groups. Specific derivatives included in this formula are clofibric acid, fenofibric acid, ciprofibrate and gemfibrozil and the like.

The chemical moiety X in the above formula is shown below.

The fibric acid analogs shown in this structure have been shown to have a large number of therapeutic applications that are quite variable and somewhat surprising. In general, these derivatives are useful in the treatment of dyslipidemia and dyslipoproteinemia. Dyslipidemia and dyslipidemia are defined here to include groups to be selected from the following; Hypercholesterolemia, abnormally elevated cholesterol levels, abnormally elevated LDL cholesterol levels, abnormally elevated total cholesterol levels, abnormally elevated plasma cholesterol levels, abnormally elevated triglyceride levels, abnormal lipoproteins Concentrations, and abnormally elevated low density lipoproteins (LDLs), abnormally elevated very low medium density lipoproteins, abnormally high density lipoprotein concentrations, hyperlipidemia, hyperchylomicronemia, abnormal levels of yeast particles Dysfunctions, and combinations thereof, which are described in The ILIB Lipid Handbook for Clinical Practice, Blood Lipids and Coronary Heart Disease, Second Edition, AM Gotto et al, International Lipid Information Bureau, New York, NY, 2000. The contents are described herein. Reference was quoted as.

Mechanism of action:

The mechanism of action of the fibrinic acid derivatives seen in clinical practice shows that activation of receptor alpha (PPAR-alpha) activated by in vivo experiments and peroxisome proliferator in transgenic mice. Has been described in in vitro experiments of human hepatocyte cultures. Through this mechanism, fibric acid derivatives are derived from plasma by activating lipolysis and lipoprotein lipase and reducing the production of apoprotein C-III (inhibitor of lipoprotein lipase activity). Increases the removal of triglyceride-rich particles.

The resulting breakdown in triglycerides produces a change in size and composition of LDL from small, dense particles (the particles are considered to be atheroogenic because of their susceptibility to oxidation) to large buoyant particles. These larger particles have greater affinity for cholesterol receptors and catalyze rapidly. Activation of PPAR-alpha also leads to an increase in the synthesis of apoproteins AI, A-II, and HDL cholesterol.

Fibric acid derivatives are also useful for the treatment of gout by decreasing serum uric acid concentrations in hyperurecemic patients.

Hyperlipidemia types include Type I, Type IIa, Type IIb, Type III, Type IV, and Type V. These types can be characterized by relative concentrations of the lipids described above (cholesterol and triglycerides) and normal concentrations of lipoproteins. Other classification methods are derived from Drug Facts and Comparisons, 52nd Edition (1998) page 1066, the contents of which are incorporated herein by reference.

Many of the fibric acid derivatives, when administered orally, do not have sufficient bioavailability and are variable, absorptive and food dependent. Indeed, the absolute bioavailability of many fibric acid derivatives is not possible because the commercially available prodrugs of fibric acid are insoluble in water, whereby parenteral forms are difficult or unavailable. Furthermore, since these agents are usually administered as esters, they are actually prodrugs. These prodrugs must be metabolized in the human body to release the active agent, which is fibric acid. However, because of the ester form of these agents, they are quite insoluble in water and are therefore difficult to formulate, nor are they easily degraded in the human body to release the active agent.

Many of the fibric acid derivatives are medium molecular weight solids with a characteristic odor. When taken orally, the drug has an unpleasant taste and can be very irritating to the mouth and throat. Taking it with food will provide higher blood levels compared to fasting. This difference in post-prandial / fasting bioavailability is more pronounced when the fibric acid derivatives are compared to their corresponding prodrug derivatives. Overall bioavailability has been reported between 40-60 and is quite variable between patients.

One of the major problems associated with commercially available fibric acid derivatives is that when the prodrugs break down in the human body, they release the prodrug portion, and the drugs themselves are very toxic. For example, in the case of fenofibrate and gemfibrozil isopropyl alcohol, an esterase enzyme is released by cleaving the promoiety from fenofibric acid. It is well known that isopropanol is highly toxic when released into mammalian tissue.

In order to improve its therapeutic efficacy, to improve blood profile, develop pharmaceutical ordered formulations, and improve the concentration of the medicament in water, the present invention overcomes some of the difficulties described above. Discusses prodrugs of alternative fibric acid derivatives.

Thus, in one aspect, the present invention relates to one alternative class of prodrugs of fibric acid derivatives. The prodrug includes hydroxyl groups of amino acids esterified with free carboxyl groups present on the fibric acid derivative molecules. In another embodiment, the amine group of the amino acid reacts with the COOH group of fibric acid to form an amide bond.

More specifically, one aspect of the present invention relates to compounds of the formula below, or a pharmaceutically acceptable salt thereof,

Where x is defined as above,

In the above formula, R is one of NH-AA or O-AA, AA is an amino acid, and one of the amine group or hydroxyl group reacts with the carboxylic acid group of the fibric acid derivative.

The invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of various of the above fibric acid derivative prodrugs and pharmaceutical carriers thereof.

In another embodiment, the invention relates to a method of treating a patient in need of fibric acid derivative therapy, the method comprising administering to the patient an effective amount of the fibric acid derivative. .

In another embodiment, the present invention relates to a method for converting a fibric acid derivative of a liquid into a solid powder by reacting one of an amino acid group or a hydroxyl functional group of an amino acid with a carboxyl functional group of a fibric acid derivative and separating their products. will be.

In another embodiment, the present invention is directed to a method that is a substantially and therapeutically effective means, comprising the preparation of derivatives that are readily absorbed upon oral administration and the continuous treatment by administering the prodrug to the patient thereby To improve the effect, and the prodrug reacts the carboxylic acid functional groups of the fibric acid derivative molecule with the amine or hydroxyl functional groups of the selected amino acids and separates their products to form ester or amide covalent bonds, respectively. And administering said product to said patient.

When naturally occurring unsubstituted amino acids are esterified to fibric acid derivatives, the resulting prodrugs are pharmaceutically ordered free flowing powders, rapidly absorbed into the body, and non-toxic as they split in the body. It has been found that it releases amino acids and requires none of the toxic emulsifiers, additives and other excipients.

Furthermore, the present invention also produced drugs, which are prodrugs of the fibric acid derivatives of the present invention; The drugs are very effective anti-hyperlipidemic drugs and have been shown to be so intact. Current amino acid prodrugs are therefore effective anti-hyperlipidemic agents and useful for the treatment of numerous high cholesterol related diseases and exhibit such potential with or without releasing effective maternal agents.

Whereas the prodrugs of the fibric acid of the present invention do not have acidic activity because of the blocking of the carboxylic acid groups, on the other hand, the prodrugs of the fibric acid are effective anti-hyperlipidemic agents with or without the release of the fibric acid derivatives. It was shown. However, all the fibric acid derivative prodrugs described above are released as in vivo experiments as effective agents having all of their pharmaceutical and cholesterol lowering properties.

Fibric acid derivative prodrugs obviously have many advantages of fibric acid derivatives, for example, all side chains cleaved from these prodrugs are naturally occurring essential amino acids and are therefore nontoxic. This leads to a high therapeutic index. Secondly, all of its prodrugs readily split into the human body to release the fibric acid derivatives. Furthermore, because of the high water solubility of the prodrugs they can be easily administered, forming an in - situ solution just prior to administration by intravenous injection with lyophilized sterile powder, or for infusion. Administration may be by providing a solution in prefilled syringes or bottles. Amino acid esters are more stable than fibric acid derivatives because the COOH groups in the fibric acid derivatives are blocking the reaction with the bases. Thus, the prodrugs of the fibric acid derivatives of the invention herein are more effective than the fibric acid derivatives themselves, which are not toxic and have no other pharmaceutical problems associated with currently marketed formulations.

The prodrugs of the present invention are useful for the treatment of hyperlipidemia in mammals whose symptoms are elevated triglycerides, low HDL (high density lipoproteins or "good" cholesterol), and elevated cholesterol. Anti-hyperlipidemic drugs. Fibric acid derivatives are also useful for reducing LDL (Low density lipoproteins, or "bad" cholesterol).

Typical examples of the synthesis of L-threonine, L-hydroxyproline and L-serine esters of fibric acid derivatives are shown in the synthetic procedures outlined below. These procedures are of course applicable to all other compounds of the fibric acid derivative classes.

Fibric acid  Derivatives Prodrugs  synthesis

The procedure for the synthesis of L-serine, L-threonine, and L-hydroxyproline esters of fenofibric acid will be outlined in the Synthesis Sequence section. These schemes are illustrative examples and can be applied equally to other amino acids. The complete process and analytical data will be provided in the experimental section. In general, fenofibric acid (100 g batches) was prepared from 4-chloro-4'-hydroxybenzophenone to correspond with the procedures in the literature. Phenofibric acid was bound to t-butyl esters of N-Boc protected amino acids (L-serine, L-threonine and L-hydroxyproline) using EDC as the binding agent and a catalytic dose of DMAP. The protecting groups were removed at low temperature with a mixture of hydrochloric acid in acetic acid (1M) with dichloromethane (5 ° C., 3-6 days). The fenofibric acid ester salts were purified by crystallization reaction from ethyl acetate and dried under high pressure vacuum conditions.

Synthetic sequence:

Synthesis of L-serine, L-threonine, and L -hydroxyproline esters of fenofibric acid : a) Boc-Ser-OtBu, EDC, DMAP, CH ₂ Cl ₂ ; b) Boc-Thr-OtBu, EDC, DMAP, CH ₂ C1 ₂ ; c) Boc-Hyp-OtBu, EDC, DMAP, CH ₂ Cl ₂ ; d) HCl, AcOH, CH ₂ Cl ₂ .

Part of the experiment:

Synthesis of SPIB00201, SPIB00202 and SPIB00203 was performed in single or two batches. Reagents mentioned in the experimental section were purchased with the highest purity available from Lancaster, Sigma, Aldrich, Acros, or Bachem, with the exception of solvents purchased from either Fisher Scientific or Mallinkrodt.

One) Fenofibric acid  synthesis

A mixture of 4-chloro-4'-hydroxybenzophenone (116 g, 0.500 mole) and sodium hydroxide (120 g, 3.00 mole) in acetone (1 L) was heated to reflux for 2 hours. The heating was stopped and the heating source was removed. A mixture of chloroform (179 g, 1.50 mole) in acetone (300 mL) was added dropwise. The reaction mixture was stirred overnight without heating. The mixture was heated to reflux for 8 hours and then allowed to cool at room temperature. The precipitate was removed by filtration and washed with acetone (100 mL). The filtrate was concentrated under reduced pressure to yield a brown oil. Water (200 mL) was added to the brown oil and acidified with IN hydrochloric acid (to pH = l). The precipitate was formed, filtered and then dried under high pressure vacuum conditions. The remaining yellow solid (268 g) crystallized from toluene in four batches (400 mL toluene each). After filtration and drying under high pressure vacuum, the experiment produced fenofibric acid as a pale yellow solid (116 g, 73% yield).

¹ H NMR (300 MHz, DMSO-d ₆ ): δ = 13.22 (1H, s, br), 7.72 (4H, d, J = 8.4 Hz), 7.61 (2H, d, J = 7.8 Hz), 6.93 (2H, d, J = 7.8 Hz), 1.60 (6H, s).

¹³ C NMR (75 MHz, DMSO-d ₆ ): δ = 192.96, 174.18, 159.35, 136.84, 136.12, 131.67, 131.02, 129.12, 128.43, 116.91, 78.87, 25.13.

2) SPIB00201  L-serine Fenofibric acid  ester

Fenofibric acid (11.6 g, 36.3 mmol), N-carbobenzyloxy-L-serine t-butyl ester (Boc-Ser-OtBu, 8.62 g, 33.0 mmol), EDC (7.59 g, 39.6 mmol), and ice Anhydrous dichloromethane (150 mL) was added dropwise to a mixture of DMAP (484 mg, 3.96 mmol) cooled in water. After the addition was completed, the ice bath was removed and the reaction mixture was stirred for 20 hours at room temperature under argon atmosphere. After 20 hours, additional dichloromethane (200 mL) was added and the solution was washed with water (2x200 mL) and brine (200 mL). It was dried over sodium sulfate, filtered and concentrated under reduced pressure. The remaining yellow oil (21.2 g) was purified by column chromatography using silica gel (400 g, 0.035-0.070 mm, 6 nm pore diameter) eluting with heptane / ethyl acetate (3: 1). After concentration under reduced pressure and drying under high pressure vacuum until the weight of the product comprising fractions became constant, the experiment was carried out with L-serine-fenofibric acid ester SPIB0020101 (16.2 g, 87 protected as a pale yellow oil). % Yield).

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 7.75 (2H, d, J = 9. 0 Hz), 7.72 (2H, d, J = 9. 0 Hz), 7.45 (2H, d, J = 8 7 Hz), 6.86 (2H, d, J = 8. 7 Hz), 5.04 (1H, d, J = 6. 9 Hz), 4.55-4.42 (3H, m), 1.66 (3H, s), 1.65 (3H, s), 1.43 (9H, s), 1.39 (9H, s).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 193.92, 172.99, 168.07, 159.24, 154.87, 138.24, 136.19, 131.94, 131.06, 130.40, 128.41, 117.26, 82.88, 80.13, 79.24, 65.44, 53.44, 28. , 25.70, 25.30.

A solution of stirred L-serine-fenofibric acid esters SPIB0020101 (16.2 g, 28.8 mmol) in anhydrous dichloromethane (100 mL) cooled to 5 ° C. under argon atmosphere was extracted with acetic acid (400 mL, 1M, 400 mmol). Hydrogen chloride solution was added dropwise. The reaction mixture was stirred at 5 ° C. for 3 days. After 3 days, the mixture was concentrated under reduced pressure and dried under high pressure vacuum to remove acetic acid. Ethyl acetate (100 mL) was added to the remaining pale yellow oil (24.7 g). The solution was concentrated and dried a second time. Ethyl acetate (65 mL) was added to the remaining pale yellow oil (17.0 g). The mixture was heated to reflux for 5 minutes and then cooled at room temperature. The precipitate was removed by filtration, dried at high pressure vacuum overnight at room temperature and then at 43 ° C. for 1 hour. The experiment produced protected L-serine-fenofibric acid ester, hydrochloride SPIB00201 (7.66 g, 60% yield) as a white solid.

¹ H NMR (300 MHz, DMSO-d ₆ ): δ = 14. 12 (1H, s, br), 8.77 (3H, s, br), 7.72 (4H, m), 7.62 (2H, d, J = 8.4 Hz), 6.92 (2H, d, J = 9.0 Hz), 4.62 (1H, dd, J = 12.0, 4.2 Hz), 4.50 (1H, dd, J = 12.0, 2.4 Hz), 4.41 (1H, m) , 1.64 (3H, s), 1.63 (3H, s).

¹³ C NMR (75 MHz, DMSO-d ₆ ): δ = 193.06, 171.70, 168.06, 158.72, 136.93, 136.06, 131.73, 131.09, 129.62, 128.49, 117.64, 79.02, 62.99, 51.11, 25.04, 24.94.

HPLC analysis: 100% purity; rt = 4.361 min .; 55% TFA (0.1%), 45% ACN; 1 mL / min; 32.3 C, Luna C18, serial # 167917-13; 20 ul inj. , NB275-49.

CHN analysis: calc. : C 54.31, H 4.79, N 3.17; found: C 54.37, H 4.78, N 3.12.

Melting Point: 151 ° C (dec.)

3) SPIB00202: L-Threonine-Phenofibric Acid Ester

  Fenofibric acid (25.5 g, 79.9 mmol), N-carbobenzyloxy-L-threonine t-butyl ester (Boc-Thr-OtBu, 20.0 g, 72.6 mmol, prepared by the method of literature), EDC (16.7 g, 87.1 mmol) and anhydrous dichloromethane (200 mL) were added dropwise to a mixture of DMAP (1.06 g, 8.71 mmol) cooled in ice water. After the addition was completed, the ice bath was removed and the reaction mixture was stirred for 20 hours at room temperature under argon atmosphere. After 20 hours, additional EDC (1.39 g, 7.26 mmol) was added and the experiment was stirred under argon atmosphere at room temperature during the day. After 4 days, additional dichloromethane (300 mL) was added and the solution was washed with water (300 mL) and brine (300 mL). It was dried over sodium sulfate, filtered and concentrated under reduced pressure. The remaining yellow oil (53.5 g) was purified by column chromatography using silica gel (500 g, 0.035-0.070 mm, 6 nm pore diameter) eluting with heptane / ethyl acetate (3: 1). After concentration under reduced pressure and drying under high pressure vacuum until the weight of the product comprising fractions became constant, the experiment was carried out with L-threonine-fenofibric acid ester SPIB0020201 (34.1 g, 82%) protected as a white foam. Yield).

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 7.74 (2H, d, J = 8.4 Hz), 7.72 (2H, d, J = 8.4 Hz), 7.45 (2H, d, J = 8.4 Hz), 6.87 (2H, d, J = 8.4 Hz), 5.47 (1H, m), 4.98 (1H, d, J = 9.9 Hz), 4,31 (1H, d, J = 9.9 Hz), 1.65 (3H, s) , 1.64 (3H, s), 1.45 (9H, s), 1.42 (9H, s), 1.22 (3H, d, J = 6.3 Hz).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 193.94, 172.14, 168.70, 159.26, 155.62, 138.28, 136.18, 131.90, 131.08, 130.37, 128.43, 117.40, 82.70, 80.17, 79.38, 72.02, 57.46, 28.30 , 26.44, 24.79, 16.90.

A solution of protected L-threonine-fenofibric acid ester SPIB0020201 (34.1 g, 59.2 mmol) in anhydrous dichloromethane (100 mL) cooled to 5 ° C. under argon atmosphere was diluted with acetic acid (600 mL, 1M, 600 mmol). Hydrogen chloride solution was added dropwise. The reaction mixture was stirred at 5 ° C. for 6 days. The mixture was concentrated under reduced pressure and dried under high pressure vacuum to remove acetic acid. Ethyl acetate (500 mL) was added to the remaining white solid (45.8 g). The mixture was heated to reflux for 10 minutes and then cooled at room temperature. The precipitate was removed by filtration and dried under high pressure vacuum conditions overnight at room temperature. The experiment produced a protected L-threonine-fenofibric acid ester, hydrochloride SPIB00202 (26.3 g, 97% yield) as a white solid.

¹ H NMR (300 MHz, DMSO-d ₆ ): δ = 14.10 (1H, s, br), 8.84 (3H, s, br), 7.73 (4H, m), 7.63 (2H, d, J = 8.1 Hz ), 6.89 (2H, d, J = 8.7 Hz), 5.44 (1H, m), 4.31 (1H, s), 1.64 (3H, s), 1.62 (3H, s), 1.38 (3H, d, J = 6.3 Hz).

¹³ C NMR (75 MHz, DMSO-d ₆ ): δ = 193.04, 171.00, 168.13, 158.76, 136.90, 136.08, 131.70, 131.06, 129.49, 128.48, 117.41, 78.99, 69.40, 55.21, 25.59, 24.22, 16.06.

HPLC analysis: 98.59% purity; rt = 4.687 min .; 55% TFA (0.1%), 45% ACN; 1 mL / min; 32.3 C, Luna C18, serial # 167917-13; 20 ul inj., NB275-49, DAD1 B, Sig = 210.4, Ref = 550,100.

CHN analysis: calc .: C 55. 27, H 5. 08, N 3.07; found: C 54.98, H 5.13, N 3.03.

Melting Point: 160.5 ℃ (dec.)

4) SPIB00203: L-hydroxyproline-fenofibric acid ester

Fenofibric acid (24.9 g, 78.1 mmol), N-carbonylbenzyloxy-L-hydroxyproline t-butyl ester (Boc-Hyp-OtBu, 20.4 g, 71.0 mmole, prepared according to the method of literature), EDC (16.3 g, 85.2 mmol), and anhydrous dichloromethane (200 mL) were added dropwise to a mixture of DMAP (1.04 g, 8.52 mmol) cooled in ice water. After the addition was completed, the ice bath was removed and the reaction mixture was stirred for 20 hours at room temperature under argon atmosphere. After 20 hours, additional EDC (1.63 g, 8.52 mmol) was added and the experiment was stirred under argon atmosphere at room temperature during the day. After 4 days, the solution was washed with water (200 mL) and brine (200 mL). It was dried over sodium sulfate, filtered and concentrated under reduced pressure. The remaining yellow oil (49.4 g) was purified by column chromatography using silica gel (500 g, 0.035-0.070 mm, 6 nm pore diameter) eluting with heptane / ethyl acetate (2: 1). After concentration under reduced pressure and drying under high pressure vacuum until the weight of the product comprising fractions became constant, the experiment was carried out with the L-hydroxyproline-phenofibrine ester SPIB0020301 (26.4 g, 63% yield).

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 7.76 (2H, d, J = 8.1 Hz), 7.73 (2H, d, J = 8.1 Hz), 7.46 (2H, d, J = 8.1 Hz), 6 . 84 (2H, d, J = 8.1 Hz), 5.32 (1H, m), 4.13 (0.38H, t, J = 7.8 Hz), 4.00 (0.62H, t, J = 7.8 Hz), 3.67 (1.62H, m), 3.46 (0.38H, d, J = 12.6 Hz), 2.29 (1H, m), 2.15 (1H, m), 1.68 (3H, s), 1.66 (3H, s), 1.44-1.38 (18H, m).

¹³ C NMR (75 MHz, CDC1 ₃ ): δ = 193.88, 172.98, 171.14, 159.25, 153.48, 138.23, 136.16, 131.99, 131.08, 130.36, 128.44, 117.03, 116.91, 81.48, 80.32, 80.20, 79.19, 74.03. , 58.23, 51.88, 51.58, 36.33, 35.31, 31.92, 28.29, 28.00, 25.89, 24.95.

Acetic acid (450 mL, 1M, 450) in a stirred solution of protected L-hydroxyproline-fenofibric acid ester SPIB0020301 (26.0 g, 44.2 mmol) in anhydrous dichloromethane (100 mL) cooled to 5 ° C. under argon atmosphere. hydrogen chloride solution was added dropwise. The reaction mixture was stirred at 5 ° C. for 4 days. After 4 days, the mixture was concentrated under reduced pressure and dried under high pressure vacuum to remove acetic acid. To the remaining yellow oil (31.5 g) was added ethyl acetate (200 mL). The mixture was sonicated, then concentrated under reduced pressure and dried under high pressure vacuum conditions. To the remaining white solid (23.2 g) was added ethyl acetate (300 mL). The ethyl acetate mixture was heated to reflux for 10 minutes and then cooled at room temperature. The precipitate was removed by filtration and dried under high pressure vacuum conditions overnight at room temperature. The experiment produced L-hydroxyproline-fenofibric acid ester, hydrochloride SPIB00203 (15.8 g, 76% yield) as a white solid.

¹ H NMR (300 MHz, DMSO-d ₆ ): δ = 14.07 (1H, s, br), 10.75 (1H, s, br), 9.40 (1H, s, br), 7.71 (4H, d, J = 8.1 Hz), 7.60 (2H, d, J = 8.1 Hz), 6.96 (2H, d, J = 8.1 Hz), 5.42 (1H, m), 4.24 (1H, t, J = 9.0 Hz), 3.61 (1H dd, J = 13.2, 4.2 Hz, 3.28 (1H, d, J = 13.2 Hz), 2.35 (2H, m), 1.66 (3H, s), 1.64 (3H, s).

¹³ C NMR (75 MHz, DMSO-d ₆ ): δ = 193.00, 171.52, 169.14, 158.81, 136.87, 136.09, 131.81, 131.05, 129.48, 128.46, 117.28, 78.99, 73.79, 57.54, 50.23, 34.13, 25.69, 24.69, 24.69, 24.69 .

HPLC analysis: 100% purity; rt = 8.369 min. ; 60% DIUF water (0.1% TFA) / 40% acetonitrile; 1 mL / min; 36.4 C; Luna Cl 8, 5u column (serial # 191070-3), 4.6x250 mm; 20 ul injection; DAD1 A, Sig = 210.4, Ref = 550,100.

HPLC-MS (ESI): calculated: M ⁺ = 431; found M + H = 432.3

Melting Point: 187.5 ℃ (dec.)

The solubility of the ester was determined in water by dissolving excess of each agent at room temperature and sinking for several hours. The resulting solutions were centrifuged at 1500 rpm for 3 minutes and the supernatant was analyzed. These esters have been shown to have solubility in water in excess of 50 mg / mL.

Experiment

Rats were checked for concentration of triglycerides in the blood over time zero. The mice then began to consume sugar, such as 30% sucrose, in water for a week. Then at the end of the week, mice were tested for triglycerides and had a normal intake. From day 7-14 mice received either experimental or control agent and tested triglycerides again on day 14 in rat blood.

In comparison of fenofibrate (control) to L-serine ester of fenofibric acid (experimental drug), three rats were tested at equivalent doses of 50, 100 and 200 mg / kg for the experimental and control groups, respectively.

The results are shown below.

Summary - studies the dosage range ferret - low dyslipidemia properties (HYPOLIPIDEMIC PROPERTY ) -Phenopy Brate and its formulation

Test substance: L-serine ester of fenofibric acid

Carrier: 1% Tween 80 in milli-Q-number

Experiment list Dosing
(Mg / Kg) Animal Number Triglycerides (mg / dl)
Day 0 7th day Day 14
Vehicle
0

One 81 168 121 2 88 171 222 3 114 133 162


Reference control fenofibrate



50

4 95 157 101 5 92 228 76 6 80 150 73 100

7 110 204 62 8 115 195 69 9 96 167 93 200

10 144 90 48 11 56 106 51 12 58 125 38


L-serine ester of fenofibric acid


50

13 88 148 86 14 94 145 86 15 100 127 73 100

16 109 - 46 17 129 100 69 18 71 183 47 200

19 74 240 83 20 81 158 61 21 42 77 46

From the above results, it can be concluded that highly water-soluble serine esters function effectively.

There are many screening experiments for determining the use of the prodrugs invented in accordance with the disclosed methods. These experiments include screening methods for both in vitro and in vivo experiments .

In vitro methods include acid / base hydrolysis of prodrugs, hydrolysis in porcine pancreas, hydrolysis in rat intestinal fluid, hydrolysis in human gastric juice, hydrolysis in human intestinal fluid, and hydrolysis in human plasma. These tests are described in Simmons, DM, Chandran, VR and Portmann, GA, Danazol amino Acid Prodrugs: In Vitro and In Situ Biopharmaceutical Evaluation, Drug Development and Industrial Pharmacy, Vol 21, Issue 6, Page 687,1995. All of which are incorporated by reference.

The prodrugs of fibric acid of the present invention are effective in treating diseases or disorders in which fibric acid derivatives are normally used. The prodrugs disclosed herein are modified in the body to release their active compounds and enhance the therapeutic advantages of fibric acid derivatives by reducing or eliminating the biopharmaceutical and pharmacokinetic barriers associated with each of them. It should be noted, however, that such prodrugs themselves will have sufficient activity without releasing any active agent in mammals.

Thus, the prodrug of the present invention enhances its therapeutic advantages by removing the biopharmaceutical and pharmacokinetic barriers of existing agents.

Furthermore, the prodrugs are easily synthesized in high yield using easy and commercially available materials.

The prodrugs of fibric acid of the present invention are effective in treating diseases or disorders in which fibric acid derivatives are normally used. The prodrugs disclosed herein are modified in the body to release their active compounds and enhance the therapeutic advantages of fibric acid derivatives by reducing or eliminating the biopharmaceutical and pharmacokinetic barriers associated with each of them. It should be noted, however, that such prodrugs themselves will have sufficient activity without releasing any active agent in mammals.

Thus, the prodrug of fibric acid of the present invention enhances its therapeutic advantages by eliminating the biopharmaceutical and pharmacokinetic barriers of existing agents. Furthermore, the prodrugs are easily synthesized in high yield using easy and commercially available materials.

It is to be understood here that in the above-mentioned formula and in the claims AA has the following definitions in the following.

AA in this definition refers to an amino acid residue having no amino groups in the main chain or side chain phase.

AA in this definition is an amino acid residue that is not a hydroxyl group on the side chain.

AA refers to an amino acid group having no carboxyl group in the main chain or side chain.

4) OAA- This is an ester bond between the hydroxy group of the agent and the carboxyl group of amino acids in the main chain or side chain. Thus, the OAA described,

Ro in this formula is a branched chain amino acid, as defined herein above.

Alternatively, it may be an ester bond between the carboxyl group of the agent and the hydroxy group on the side chain of amino acids having hydroxy groups such as threonine, serine, hydroxyproline, tyrosine and the like. The hydroxy group forms part of the ester bond represented by O. Thus, as described, AA refers to an amino acid having a hydroxy group on the side chain, but as with OAA indicated above, AA does not have a hydroxy group because the oxygen atom is represented in the formula.

While the invention has been described in connection with the description thereof, it should be understood that the foregoing description is intended to be illustrative of the scope of the invention and not of limitation, but defined by the scope of the appended claims. Other aspects, advantages and modifications are within the scope of the following claims.

Claims (1)

A pharmaceutical composition comprising a therapeutically effective amount of the formula or a pharmaceutically acceptable salt thereof:

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