JP2013035863A - Amino acid prodrug - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a prodrug comprised of an amino acid bonded to a medicament or drug having a hydroxy, amino, carboxy or acylating derivative thereon.SOLUTION: The invention is directed to the prodrug having the same utility as the drug from which it is made, but having enhanced therapeutic properties. The drug or medicament useful contains functional groups thereon that permit the drug to react with and form a covalent bond with the amino acid. Examples of functional groups present on the drugs include NH2, OH, COOH or acid derivatives thereof, such as esters, amides and the like.

Description

The present invention relates to amino acid derivatives of pharmaceutical compounds, methods of treating specific diseases, which are improved by the administration of these drugs and pharmaceutical compositions containing these drugs. The present invention encompasses many improved physiochemical, biological and clinical effects of various agents using amino acids as covalent carriers.

The development of compounds to treat disorders, diseases and illness is becoming increasingly difficult and expensive. The probability of successful development is low enough to be discouraged. Furthermore, the development time can be close to 10 years or more, and many patients are left untreated for long periods of time.

Even when effective pharmaceutical compounds are developed, there are a number of drawbacks associated with their administration. Such drawbacks include sensory and pharmacokinetic disorders that adversely affect the effects of existing pharmaceutical compounds. For example, an unpleasant taste or odor of a pharmaceutical compound or composition can be a serious obstacle to patient compliance in a dosing regimen. Problems with the solubility of pharmaceutical compounds can make it difficult to prepare a uniform composition. Other drawbacks associated with known pharmaceutical compounds include: poor absorption of oral dosage forms; low bioavailability of pharmaceutical compounds in oral dosage forms; low stability of pharmaceutical compounds in vitro and in vivo; dose proportionality Lack of blood-brain barrier; excessive first-pass metabolism of pharmaceutical compounds when passing through the liver; excessive enterohepatic recirculation; low absorption rate; inefficient compound release at the site of action; For example, gastrointestinal non-irritant and / or ulceration; pain upon infusion of parenterally administered pharmaceutical compounds and compositions; some pharmaceutical compounds and compositions require excessive dosages, and other undesirable Characteristics. Some pharmaceutical compounds produce toxic by-products that are processed in the body with harmful effects.

The art continues to seek new compounds with improved properties that overcome the shortcomings of the known pharmaceutical compounds described above for treating a variety of disorders.

The present invention overcomes many of the problems associated with currently marketed drugs by making prodrugs. The concept of prodrugs is well known, and the literature lists many examples of such prodrugs, including many groups including various groups such as statins, ACE inhibitors, and antiviral drugs such as acyclovir. Prodrugs are commercially available. However, the present invention uses amino acids as a part for producing a prodrug. The prodrugs of the present invention have many advantages. For example, amino acid prodrugs are administered by various routes such as oral, IV (intravenous), rectal, or other methods, and these prodrugs are converted to active drug molecules. A major advantage of amino acid prodrugs is that they are non-toxic and are therefore absorbed into the body or excreted safely. This is due to the toxicity of the promoty itself, as is the case with the highly toxic statin drugs, ACE inhibitors such as enalapril and benazapril, and many antibiotics such as pivoxil, isopropyl, axetyl and cilexetil. This is a feature not found in commercially available prodrugs that reduce the therapeutic index of the active agent.
On the other hand, the amino acid prodrug of the present invention also provides many advantages as shown below.

SUMMARY OF THE INVENTION The present invention is a pharmaceutically active prodrug having an amino acid covalently linked to a pharmaceutical compound that forms the acidic prodrug, and is in a form such as a mammal. Relates to a prodrug administered in

式中Ｒ０はアミノ酸の側基または側鎖である。本明細書に定義される

は、アミノ酸の主鎖である。従って、例えば、主鎖上のアミノ基およびカルボキシル基以外に、側鎖はその中に官能基を有してよい。それは、アミノ酸と薬剤との間の供給結合を可能にする、アミノ酸部分中の官能基である。 Amino acids are ideal models used as prodrugs because they can make various types of bonds between themselves and drugs. By definition, an amino acid has at least two functionalities therein: an amino group (NH ₂ ) and a carboxyl group (COOH). For example, α-amino acid has the following known structure:

In the formula, R0 is a side group or side chain of an amino acid. Defined herein

Is the main chain of the amino acid. Thus, for example, in addition to amino and carboxyl groups on the main chain, the side chain may have functional groups therein. It is a functional group in the amino acid moiety that allows a supply bond between the amino acid and the drug.

Agents or medicaments useful in the present invention contain functional groups therein that allow the agent to react with an amino acid to form a covalent bond. Examples of functional groups present in the _drug, NH 2, OH, COOH or esters, their acid derivatives amide.

Addition forms of pharmaceutical compounds and amino acids can be realized through the following bonds.
1) an ester bond (—CO—O—) by condensation of a carboxylic acid with an alcohol or phenol hydroxyl group or through transesterification, for example
a) If the pharmaceutical compound has an aliphatic or aromatic hydroxyl group, an ester bond can be formed with the backbone carboxylic acid group of the amino acid under esterification conditions.
b) If the pharmaceutical compound has an aliphatic or aromatic hydroxyl group and the amino acid has a side chain carboxylic acid group, ester bonds can be formed between them under esterification conditions.
c) If the pharmaceutical compound has a carboxylic acid group and the amino acid has a side chain aliphatic or aromatic hydroxyl group, ester bonds can be formed between them under esterification conditions.
d) the pharmaceutical compound has an ester group with a substituted or unsubstituted acyloxy (eg alkoxy- or arylalkoxy-, aryloxycarbonyl) substituent (compound-O-CO-substituent) and the amino acid is a backbone carboxylic acid If they have groups, ester bonds can be formed between them via transesterification.
e) The pharmaceutical compound has an ester group with a substituted or unsubstituted acyloxy (eg alkoxy- or arylalkoxy-, aryloxycarbonyl) substituent (compound-O-CO-substituent), and the amino acid is a side chain carboxylic In the case of having acid groups, ester bonds can be formed between them via transesterification.
f) the pharmaceutical compound has an ester group with a substituted or unsubstituted alkoxy or arylalkoxy or aryloxycarbonyl substituent (compound-CO-O-substituent) and the amino acid is a side chain aliphatic or aromatic hydroxyl group Ester bonds can be formed between them via transesterification.
g) The alcohol and carboxylic acid moieties may be on the same molecule so that they can form internal esters. For example, compounds capable of forming an internal ester under ester forming conditions such as gabapentin are also included in the scope of the present invention.
2) Amide bond (-CO-NH-) resulting from condensation of carboxylic acid and amine, for example
a) If the pharmaceutical compound has an amino group and the amino acid has a backbone carboxylic acid group, an amide can form under amide forming conditions.
b) If the pharmaceutical compound has an amino group and the amino acid has a side chain carboxylic acid group, an amide bond can form between them under amide forming conditions.
c) If the pharmaceutical compound has a carboxylic acid group and the amino acid has a backbone amino group, an amide bond can form between them under amide forming conditions.
d) If the pharmaceutical compound has a carboxylic acid group and the amino acid has a side chain amino group, an amide bond can form between them under amide forming conditions.

Accordingly, the present invention is directed to prodrugs thus formed. As shown below, the prodrugs formed in this way have advantages that cannot be achieved with drugs that have no added amino acids. For example, it can improve bioavailability, efficacy, lower toxicity, show higher water solubility, and / or increase the ability of the drug to cross the cell membrane or blood brain barrier and have fewer side effects such as gastrointestinal irritation , Increase the therapeutic index, etc.

Thus, the present invention is directed to a method for improving the quality of drug treatment. Here, the improvement in the quality of treatment is improved efficacy, increased therapeutic index, increased solubility in mammalian body fluids, cell membrane permeability compared to the corresponding drug administered to the patient in the form of a non-prodrug. Selected from the group consisting of an improvement in blood pressure, an improvement in blood-brain barrier permeability, a decrease in side effects such as irritation and / or a significant decrease in ulceration, a decrease in toxicity, an increase in absorption rate, etc. And a covalent bond is formed between them, and the product (hereinafter referred to as “prodrug”) is administered to the patient. The prodrugs of the present invention have at least one improved characteristic. In practice, it is preferred that they have at least the two improved characteristics described above. Other advantages of prodrugs include the wide availability of amino acids and ease of reaction. Reactions that form amides are generally efficient and their yields are extremely high, greater than about 70%, more preferably greater than about 80%, and most preferably greater than about 90%.

DETAILED DESCRIPTION OF THE INVENTION As used herein, the terms "drug", "pharmaceutical" and "pharmaceutical" are used interchangeably and are activities administered to a patient without the addition of amino acids. Represents a compound. Furthermore, as used herein, the agent includes can react with amino acids thereon, NH 2, _OH, COOH, or acylated derivatives thereof (such as esters, anhydrides, amides) a functional group such as. Where the drug is linked to an amino acid, the term “amino acid prodrug” or “prodrug of the invention” or a synonym thereof is used.

Some amino acids useful as promoties (ie, react with drugs) include the free amino and / or carboxylic acid groups of all common amino acids. Among them, some preferred embodiments are at least 100 g / L, more preferably at least 250 g / L, even more preferably at least 500 g / L for deionized water in an unbuffered aqueous solution at 25 ° C. Includes amino acids with relatively high solubility of 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 a promoiety are amino acids containing basic amino side chains such as lysine. For example, lysine has a solubility of about 700 g / L in deionized water at 25 ° C.

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

Another preferred embodiment is at most 10 g / L, or at most 2 g / L, or at most 0.6 g / L in deionized water at 25 ° C., where the solubility of the aqueous medium is relatively low. Includes certain amino acids. For example, the solubility of tyrosine in deionized water at 25 ° C. is about 0.5 g / L. Such prodrugs can be used to produce formulations with long-term release characteristics due to the limited solubility of prodrugs.

Among the other amino acids useful as promoies are amino acids containing carboxylic acid side chains such as glutamic acid and aspartic acid. Other amino acids useful as promoies are non-essential amino acids and non-natural amino acids.

またＲ_１は、この場合Ｒ_０が以下列挙するアミノ酸の側鎖である。 The following reaction scheme represents the reaction described above for the reaction of drugs including hydroxyl, carboxyl, and amine with various amino acids. In the scheme below, R is a drug with few functional groups, whether any of OH, COOH or NH ₂ functional groups are present,

In addition, R ₁ is a side chain of the amino acid R ₀ listed below in this case.

Reaction scheme A: A scheme in which a hydroxyl group of a drug reacts with a carboxyl group of an amino acid to form an ester prodrug.

Reaction Scheme B: A scheme in which a carboxyl group of a drug reacts with a hydroxyl group of a hydroxy amino acid having a hydroxy group on its side chain to form an ester prodrug.

Reaction scheme C: A scheme in which an amino group of a drug reacts with a carboxyl group of an amino acid to form an amide prodrug.

Reaction scheme D: A scheme in which a carboxyl group of a drug reacts with a carboxyl group of an amino acid to form an anhydride prodrug.

Reaction scheme E: A scheme in which an amine group of a drug reacts with an amine group of an amino acid to form an azoprodrug derivative.

Reaction scheme F: A scheme in which a carboxyl group of a drug reacts with an amine group of an amino acid to form an amide prodrug.

In the above schemes A to 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 salt thereof therein. In solution, these two end groups ionize and double ionize through a neutral state that is recognized as a zwitterion. The amine donates electrons to the carboxyl group and its ionic end group is stabilized in aqueous solution by polar water molecules.

It is the side group that distinguishes amino acids from each other. Some amino acids such as lysine have an amino group on the side chain. In addition, amino acids such as threonine, serine, hydroxyproline and tyrosine have a side chain containing a hydroxy group. Some amino acids such as glutamic acid or aspartic acid have a carboxy group on the side chain. These functional groups on the side chains can also form covalent bonds with drugs such as esters and amides. When these side groups, such as hydroxy groups, are included in such linkages, the bond is represented as OAA, where AA has a side chain with a hydroxy group, but the hydroxy group It is an amino acid residue that does not have. That is, AA according to this definition is an amino acid having no hydroxy side group because the hydroxy group participates in the ester formation reaction. Furthermore, when an ester is formed between the hydroxy group of an amino acid and the OH group of the drug, the hydroxy group on the carboxy group forms a byproduct with the hydrogen of the hydroxy group, so that the resulting product is a carboxy group. Does not have OH in the group, but only has an acyl moiety. When the bond is represented by C (= O) -NHAA, it means that the amino acid is formed as an amide bond between the carboxy group of the drug and the amino group of the amino acid. However, as described, amide bond NH is derived from an amino acid, so AA is an amino acid having no amino group.

A preferred amino acid is a natural amino acid. The amino acid is more preferably an α amino acid. It is also preferred that the amino acid is in the L-configuration. Preferred amino acids include 20 essential amino acids. Preferred amino acids include lysine (Lys), leucine (Leu), isoleucine (Ile), glycine (Gly), aspartic acid (Asp), glutamic acid (Glu), methionine (Met), alanine (Ala), and valine (Val). , Proline (Pro), histidine (His), tyrosine (Tyr), serine (Ser), norleucine (Nor), arginine (Arg), phenylalanine (Phe), tryptophan (Trp), hydroxyproline (Hyp), homoserine (Hsr) ), Carnitine (Car), ornithine (Ort), canavanine (Cav), asparagine (Asn), glutamine (Gln), carnosine (Can), taurine (Tau), diencoric acid (Dik), γ-aminobutyric acid (GABA) , Cysteine Cys), cystine (Dcy), sarcosine (Sar), include threonine (Thr) and the like. Further preferred amino acids are the 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. It is.

Prodrugs are prepared from drugs that have groups on them that can react with amino acids.

Preferred agents for reacting with amino groups according to various schemes are as follows:

The prodrugs of the present invention contain amino groups, which are naturally basic. They can form a wide variety of pharmaceutically acceptable salts with various inorganic and organic acids. These acids which can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are non-toxic acid addition salts, ie hydrochlorides, hydrobromides, hydroiodic acids. Form salts containing pharmaceutically acceptable anions such as salts, nitrides, sulfates, bisulfates, phosphates, formates, acetates, citrates, tartrate, lactates and the like.

As shown herein, in one embodiment, the present invention provides for esterification of, for example, cyclosporine and MeBmt (xy = CH═CH), or dihydro MeBmt moieties (xy = CH ₂ CH ₂ ). Prodrugs containing the amino acids prepared are intended. Amino acids are added to cyclosporine and other drugs by covalent bonds.

The compounds of the present invention are prepared by techniques recognized in the art. For example, when the drug contains an OH group like cyclosporine, an amino acid or an acylated derivative thereof, for example, an acid halide such as amino acid fluoride or amino acid chloride, or an alkyl group contains 1 to 6 carbon atoms An amino acid alkyl ester or the like reacts with a carboxy group of a drug such as cyclosporine under the conditions of esterification. Preferably, the reaction is carried out in the presence of an acid such as hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid and the like. Alternatively, as described above, when the drug has an amino group, the amino acid reacts with the drug under conditions of amide formation to form an amide as a covalent bond. Alternatively, if the drug has a carboxy group or an acylated derivative, the drug reacts with the amino group of the amino acid to form an amide under amide forming conditions, forming an amide bond between the amino acid and the drug. In addition, if the drug has a carboxy group therein, the side chain hydroxy group of the amino acid reacts with the carboxy group or the acylated derivative in which, under esterification conditions, between the amino acid and the drug as described above. An ester linkage can be formed.

If the amino acid has a group that is reactive under the reaction conditions, the group is protected by protecting groups known in the art. After the reaction is complete, the protecting group is removed. Examples of protecting groups that can be used include Theodora W. et al. It is described in a book entitled “Protective Group in Organic Synthesis” John Wiley & Sons, 1981 by Greene, the contents of which are incorporated by reference.

For example, when an amino acid having a carboxyl group in its side chain, such as aspartic acid and glutamic acid, is used for the above synthesis, it is generally necessary to protect the side chain carboxyl group. Suitable protecting groups are cyclohexyl esters, t-butyl esters, benzyl esters, allyl esters and the like, 9-fluorophenyl-methyl groups, or 1 which can be protected after completion of the esterification reaction using techniques known to those skilled in the art. -Or adamantyl group such as 2-adamantyl.

For example, when amino acids having a hydroxyl group in its side chain, such as serine, threonine, hydroxyproline, and the like, and amino acids having a phenol group in its side chain, such as tyrosine, are used in the above esterification and reaction, preferably These side chain hydroxyl groups or side chain phenol groups need to be protected. A suitable protecting group for the hydroxyl side group is an ether such as benzyl ether or t-butyl ether. Benzyl ether can be removed using liquid hydrogen fluoride, while t-butyl can be removed by treatment with trifluoroacetic acid. Suitable protecting groups for the phenol side groups may be ethers as described above including benzyl or t-butyl ether, or 2,6-dichlorobenzyl, 2-bromobenzyloxycarbonyl, 2,4-dinitrophenyl, and the like.

Further, the product can be purified and substantially purified using techniques known to those skilled in the art, such as chromatography such as HPLC, crystallization and the like. Substantially “pure” means that the product contains no more than about 10% impurities.

Prodrugs can be prepared using pharmaceutically acceptable solvents, esters, enantiomers, diastereomers, N-oxides, polymorphic prodrugs or pharmaceuticals thereof using one of the techniques known to those skilled in the art. Pharmaceutical compositions can be made that contain a pharmaceutically acceptable carrier, and optionally but desirably a pharmaceutically acceptable excipient, together with a pharmaceutically acceptable salt.

The prodrug utilized in the method is used in a therapeutically effective amount.

The physician will determine the optimal dose of the prodrug of the present invention, which will vary depending on the selected mode of administration and the particular compound, and further includes, but is not limited to, the patient being treated, It will vary depending on various factors, including the age of the patient, the severity of the condition being treated, etc., and the uniqueness of the prodrug administered. The physician generally desires to begin treatment at a dose significantly less than the optimal dose of the compound and gradually increase the dosage until the optimal effect is obtained in that environment. In general, it can be seen that when the composition is administered orally, a larger amount of active agent is required to achieve the same effect as compared to a smaller amount given parenterally. The compounds are as useful as the non-persistent counterparts and dosage levels are comparable to those commonly used with other therapeutic agents. When given parenterally, the compound is usually administered in an amount of about 0.001 to about 10,000 mg / kg body weight, depending on the host and the severity of the condition being treated and the compound used. It depends.

In a preferred embodiment, the compound used is from about 0.01 mg / kg to about 1000 mg / kg body weight, more preferably from about 0.1 to about 1000 / kg body weight, depending on the particular mammalian host or the disease being treated. An amount in the range of 500 mg / day is administered orally. This dosage regimen may be adjusted by the physician to provide the optimal therapeutic response. For example, the dose can be divided into multiple doses administered daily, or the dose can be reduced depending on the urgency of the treatment condition.

Prodrugs may be administered by any conventional method, such as oral, intravenous, intramuscular or subcutaneous routes.

The prodrug may be administered, for example, with an inert diluent, or an anabolic edible carrier, or enclosed in a hard or soft shell gelatin capsule, compressed into tablets, or a meal It may be incorporated directly into the food. For oral therapeutic administration, prodrugs may incorporate excipients and may be used in the form of ingestible tablets, buccals, troches, capsules, elixirs, suspensions, syrups, cachets and the like. Such compositions and preparations should contain at least 1% prodrug. The proportions of the compositions and preparations can of course vary and usually will be between about 5 and about 80% of the unit weight. The amount of prodrug used in such therapeutic compositions is such that a suitable dosage will be obtained. Preferred compositions or preparations of the present invention contain about 200 mg to about 4000 mg of prodrug. Tablets, troches, pills, capsules and the like may include: Binders such as tragacanth gum, acacia, corn starch or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch, potato starch, alginic acid, lubricants such as magnesium stearic acid, and sucrose, lactose or saccharin Sweeteners or flavorings such as peppermint, wintergreen oil, or cherry flavor may be added. When the dosage unit form is a capsule, it may contain a liquid carrier in addition to the types of materials described above.

It may be coated with various other materials, or the physical shape of the dosage unit may be altered. For instance, tablets, pills, or capsules may be coated with shellac, sugar or both. A syrup or cachet may contain the active compound, sucrose as a sweetening agent, methyl or propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, the materials used in any dosage unit form preparation must be pharmaceutically pure and substantially non-toxic in the amounts used. In addition, the active compound can be incorporated into sustained-release preparations or formulations. For example, as a sustained release dosage form, the active ingredient is optionally bound to an ion exchange resin, which can optionally be provided with a diffusion barrier coating to alter the release characteristics, or the prodrug of the present invention. A form associated with a sustained-release polymer known in the art, such as hydroxypropylmethylcellulose, can be considered.

Prodrugs can also be administered parenterally or intraperitoneally. It is particularly advantageous to prepare parenteral compositions in a uniform dosage form that is easy to administer. Dispersants can also be prepared in glycerol, eg polyethylene glycol solutions such as PEG 100, PEG 200, PEG 300, PEG 400, and mixtures thereof, and oils. Under normal storage and use conditions, these preparations contain a preservative to prevent the growth of microorganisms.

Pharmaceutical forms suitable for use by injection include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectables or dispersions. In either case, the shape is usually sterile and requires fluidity to be injected. It needs to be stable in the state of manufacture and storage and must usually be protected from the contaminating action of microorganisms such as bacteria and fungi. The carrier can be, for example, water, ethanol, polyol (for example, glycerol, propylene, propylene glycol, and one or more liquid polyethylene glycols, for example, as disclosed herein), suitable mixtures thereof, and vegetable oils It may be a solvent or dispersion medium containing The proper fluidity can be maintained, for example, by applying a coating such as lecithin, maintaining the required particle size in the case of a dispersant, and using a surfactant. Microbial activity can be prevented by various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be desirable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by using an absorption delaying agent, for example, aluminum monostearate, in the composition.

A sterile injectable solution is prepared by incorporating a necessary amount of a prodrug in an appropriate solvent together with various other components as necessary, followed by filter sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle that contains the basic dispersion medium and the required of the other ingredients mentioned above. In the case of a sterile powder, the above solution is vacuum dried or lyophilized as required.

Prodrugs can also be applied topically, eg through patches, using techniques known to those skilled in the art. Prodrugs can be administered orally using methods known to those skilled in the art after preparing a suitable formulation of the prodrug of the present invention. These formulations are prepared with suitable non-toxic pharmaceutically acceptable ingredients. These ingredients are known to those skilled in the preparation of buccal dosage forms. Some of these components can be found in Remington's Pharmaceutical Sciences, 17th edition (1985), a standard reference material in this field. The selection of a suitable carrier is highly dependent on the exact nature of the desired buccal dosage form, such as tablets, lozenges, gels, patches, and the like. All these buccal dosage forms are considered to be within the scope of the present invention and they are prepared in a conventional manner.

The formulation of the pharmaceutical composition may be prepared by conventional methods using one or more physiologically and / or pharmaceutically acceptable carriers or excipients. Thus, the compounds and their pharmaceutically acceptable salts and solvents may be prepared for administration by inhalation or insufflation (via the mouth or nose) or oral, buccal, non-oral or rectal administration. For oral administration, the pharmaceutical composition comprises, for example, a binder (eg, pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropylmethylcellulose), a filler (eg, lactose, microcrystalline cellulose, or calcium hydrogen phosphate), Pharmaceutically acceptable excipients such as lubricants (eg magnesium stearate, talc or silica), disintegrants (eg potato starch, or sodium starch glycolate), or wetting agents (eg sodium lauryl sulfate) May be used in the form of tablets or capsules prepared by conventional methods. The tablets may be coated by methods known in the art.

Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or may be provided as a dry product for constitution with water or other suitable vehicle before use. Such liquid formulations include suspensions (eg, sorbitol syrup, corn syrup, cellulose derivatives or edible hardened oils and fats), emulsifiers (eg, lecithin or acacia), non-aqueous vehicles (eg, almond oil, oily esters, Ethyl alcohol or fractionated vegetable oils) and pharmaceutically acceptable additives such as preservatives (eg, methyl or propyl p-hydroxybenzoate or sorbic acid) may be prepared by conventional means. The preparation may contain buffer salts, flavoring agents, coloring agents and sweeteners as required. Formulations for oral administration can be formulated appropriately and given active prodrugs in a controlled manner.

The prodrugs of the present invention may be formulated for parenteral administration, eg, by injection such as bolus injection or continuous infusion. Injectable preparations may be presented in unit dosage forms, such as capsules with preservatives, or multidose containers. The compositions may take the form of oily or aqueous vehicle suspensions, solutions or emulsions and may contain formulations such as suspending, stabilizing and / or dispersing agents. Alternatively, the prodrug may take the form of a powder for constitution with a suitable vehicle, such as sterile pyrogen-free water, before use.

The prodrugs of the present invention may be formulated in rectal compositions such as suppositories or indwelling enemas, eg, containing conventional suppository bases such as cocoa butter or other glycerides.

In addition to the formulations described above, the prodrugs of the present invention may be formulated as a depot preparation. Such long acting formulations can be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, prodrugs may be formulated with suitable polymers or hydrophobic materials (eg, as acceptable oil emulsions) or ion exchange resins, or as poorly soluble derivatives, such as, for example, poorly soluble salts. .

Pharmaceutical compositions containing the prodrugs of the present invention may be given in packs or dispenser devices containing one or more unit dosage forms containing the active ingredients, as appropriate. The pack may wrap, for example, a metal or plastic foil such as a blister pack. The pack or dispenser device is accompanied by instructions for administration.

In tablet form, it is desirable to include a lubricant that facilitates the manufacturing process of the dosage unit. Lubricants also optimize dissolution rate and drug flow. When present, the lubricant is present from about 0.01% to about 2%, preferably from about 0.01% to about 0.5% by weight of the dosage unit. Suitable lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, stearyl fumaric acid, talc, hydrogenated vegetable oil, and polyethylene glycol. As will be apparent to those skilled in the art, changes in the particle size and / or unit density of the components within the dosage unit may have similar effects--improved manufacturability and dissolution rate and drug flow without the addition of a lubricant. Gender optimization.

Optionally other ingredients may be incorporated into the dosage unit. Such additional optional ingredients include, for example, one or more disintegrants, diluents, binders, enhancers, and the like. Examples of disintegrants that can be used include cross-linked polyvinyl pyrrolidone such as crospovidone (eg, polyplastidone (registered trademark) XL available from GAF), croscarmellose (eg, Ac-Di-sol (available from FMC) Cross-linked carboxymethylcellulose, alginic acid and sodium carboxymethyl starch (eg, Explotab® available from Edward Medcell Co., Inc.), agar bentonite and alginic acid such as, but not limited to. Suitable diluents include those generally useful for pharmaceutical formulations prepared using compression techniques, such as: dicalcium phosphate dihydrate (eg, available from Stauffer) i-Tab (registered trademark)), sugar crystallized with dextrin (for example, co-crystal of sucrose and dextrin such as Di-Pak (registered trademark) available from Amstar), calcium phosphate, cellulose, kaolin , Mannitol, sodium chloride, dry starch, powdered sugar, etc. Binders, when used, enhance adhesion, such as starch, gelatin, and sucrose, dextrose, Examples include, but are not limited to, sugars such as molasses and lactose New dosage units may contain permeation enhancers to increase the rate of passage of the active ingredient into the oral mucosa. Examples include dimethyl sulfoxide (“DMSO”), dimethylformamide (“DMF”), N, N-dimethylacetate. Amide ( "DMA"), decylmethylsulfoxide ( _"C 10 MSO"), polyethylene glycol monolaurate ( "PEGML"), glycerol monolaurate, lecithin, the 1-substituted azacycloheptan-2-ones, particularly 1 N-dodecylcyclazacycloheptan-2-one (available from Nelson Research & Development Co., Irvine (California) under the trademark Azone.RTM.), Lower alkanols (eg ethanol), SEPA® (Macrochem) Co., Lexington (Massachusetts)), cholic acid, taurocholic acid, bile salt type enhancer, Tergitol (R), Nonoxynol-9 (R) and TW Surfactants such as EEN-80 (registered trademark) can be mentioned, but are not limited thereto.

Optionally, flavoring agents may be included in various pharmaceutical formulations. Any suitable flavoring agent may be used, such as artificial sweeteners such as mannitol, lactose or aspartame. Coloring agents may be added, but such agents are not necessary. Examples of colorants include any water soluble FD & C dye, mixtures thereof, or their corresponding lakes.

In addition, if necessary, the dosage unit can be formulated with one or more preservatives, or bacteriostatic agents such as methylhydroxybenzoate, propylhydroxybenzoate, chlorocresol, benzalkonium chloride, and the like.

As used herein, “pharmaceutically acceptable carrier” refers to any solvent, dispersion medium, coating agent, antibacterial and antifungal agent, etc. for pharmaceutically active substances known in the art, etc. Contains tonicity as well as absorption retardants. Unless conventional media or agents are incompatible with the prodrug, they are contemplated for use in therapeutic compositions. Supplementary active ingredients can also be incorporated into the compositions.

As used herein, dosage unit form means a physically discrete unit suitable as a unit dose for the subject to be treated. Each unit contains a predetermined amount of a prodrug, calculated to produce the desired therapeutic effect, with the required pharmaceutical carrier.

Prodrugs are prepared in the dosage unit form described above so that an effective amount can be conveniently and effectively administered with a suitable pharmaceutically acceptable carrier. The amount of principal active compound contained in a unit dose ranges, for example, from about 10 mg in humans, or as little as 1 mg (for small animals) to about 2000 mg. When in solution, the concentration of the prodrug is preferably in the range of about 10 mg / mL to about 250 mg / mL. In the case of a composition containing supplementary active ingredients, the dosage is determined with reference to the usual dose and the mode of administration of said ingredients. For buccal administration, the prodrug is preferably present in an oral unit dosage form in an amount ranging from about 10 to about 50 mg.

The prodrugs of the present invention are effective in the treatment of diseases or conditions in which the corresponding drug (not including the amino acid prodrug of the present invention) is commonly used.

As used herein, the term “treating” reverses, alleviates or prevents the progression of the disease, disorder or condition in which the term is used, or one or more symptoms of such disease, disorder or condition. Means that. As used herein, “treating” also compares the likelihood or incidence of a mammal's disease, disorder, or condition relative to an untreated control population or to the same mammal prior to treatment. It also means lowering. For example, as used herein, “treating” may mean preventing a disease, disorder or condition, delaying or preventing the occurrence of the disease, disorder or condition, or disease. It may include delaying or preventing the onset of symptoms associated with a disorder or condition. As used herein, “treating” further reduces the severity of the disease, disorder or condition, or symptoms associated with the disease, disorder or condition before the mammal suffers from the disease, disorder or condition. It also means. Preventing or reducing the severity of a disease, disorder, or condition before such distress occurs can cause a subject who is not suffering from the disease, disorder, or condition at the time of administration to a subject of the invention described herein. It relates to administering the composition. As used herein, “treating” also means preventing the recurrence of a disease, disorder or condition, or the recurrence of one or more symptoms associated with such a disease, disorder or condition. The terms “treatment” and “therapeutically” refer to the act of treating as “treating” is defined above.

As used herein, the term “patient” or “subject” refers to warm-blooded animals, preferably mammals such as primates, including, for example, cats, dogs, horses, cows, pigs, mice, rats and humans. means. The preferred patient is a human.

The prodrugs of the present invention show similar utility as the corresponding drugs without amino acid linkages. Prodrugs exhibit high therapeutic properties. That is, they exhibit at least one, and more preferably at least two, high therapeutic properties compared to drugs that were not converted to the prodrugs of the invention prior to administration. Such therapeutic properties include, but are not limited to, the following:
a. Improved taste, smell b. Desirable octanol / water partition coefficient (ie solubility in water / fat)

The solubility of various amino acids in aqueous solutions varies. By selecting specific amino acids, the octanol water partition coefficient is affected. For example, many drugs in the list below have high hydrophobicity. Amino acids are also highly hydrophobic. For example, assume that the drug is propofol and the amino acid is lysine. Propofol is completely insoluble in water, while ricin has a solubility of up to 700 mg / ml. When these two different molecules are esterified with an ester bond, the resulting lysine ester of propofol exhibits a solubility in water of greater than 250 mg / ml.

On the other hand, cromolyn sodium has high water solubility. For any practical use, cromolyn sodium is not absorbed when administered orally. Absorbency can be improved by changing its water solubility. In this case, a state opposite to that of propofol is obtained. That is, the ultimate goal is to reduce water solubility. A suitable hydrophilic / lipophilic balance can be obtained by selecting an appropriate amino acid having low water solubility such as tyrosine.
c. Improving stability in vitro and in vivo d. Increased blood-brain barrier passage e. Removal of the first-pass effect in the liver, i.e. the drug is not metabolized in the liver, so that more drug circulates in the line f. Reduction of enterohepatic recirculation (this improves bioavailability)
g. Painless injection of parenteral preparations h. Improved bioavailability i. Improved change in absorption rate (its increase vs. deficiency)
j. Reduction of side effects k. Dose proportionality

Dose proportionality requires that a proportionally increased amount of active agent be delivered into the blood stream when administered in increasing doses of the drug. This is measured by determining the area under the plasma concentration versus time curve obtained after administration of the drug via a route other than the IV route and measuring by measuring the drug in plasma / blood. A simple mathematical operation is as follows. For example, the drug is orally administered to a patient, for example, at three different doses, 10 mg, 100 mg and 1000 mg, and the area under the plasma concentration time curve (AUC) is measured. Each total AUC is then divided by the dose and the result must be the same for all three doses. In that case there is a dose proportionality. The lack of dose proportionality indicates that one or more pharmacokinetic / pharmacological mechanisms are saturated, including the number of receptor sites available for absorption, metabolism, or pharmacological response.

For example, in the above study, if the AUC values are 100,000 and 10,000, the dose proportionality is inappropriate in this example. In the absence of dose proportionality, the amount of drug in the plasma is either higher or lower, depending on the saturation mechanism. Here are some possibilities: Saturable absorption. In this case, as the dose is increased, the amount of drug absorbed decreases proportionally, and thus the overall AUC decreases with increasing dose.

Saturation of eliminated metabolism. In this case, more drug circulates in the blood and AUC increases with increasing dose.

Saturation of the pharmacological receptor site. In this case, all receptor sites are eventually occupied by the drug, so adding the drug does not increase the response. Thus, increasing the dose does not result in an increased response.

Dose proportionality is an excellent response profile because it can accurately predict pharmacological response and healing power for all doses. Thus, dose proportionality is a desirable property for any drug. Furthermore, achieving dose proportionality also depends on the formulation and the difference in feeding / fasting.

l. Selective hydrolysis of prodrugs at the site of action m. Controlled release characteristics n. Targeted drug delivery o. Reduce toxicity, ie improve treatment rate p. Dose reduction q. Changes in metabolic pathways that deliver larger amounts of drug to the site of action r. Increased aqueous solubility s. Increased efficacy

Thus, there are a variety of possible dosage forms for amino acid prodrugs, which are prepared by the following conventional methods.
i. Oral liquid dosing (including chewable tablets, sugar-containing and sugar-free, color-containing and non-colored, controlled-release and immediate-release solutions containing alcohol-containing and alcohol-free preparations)
ii. Oral solid dosage (controlled release and immediate release tablets, capsules and caplets)
iii. Intravenous (both injection, prepared, and lyophilized powder)
iv. Intramuscular (both injection, prepared, and lyophilized powder)
v. Subcutaneous (both injection, prepared, and lyophilized powder)
vi. Transdermal (mainly patches)
vii. In the nose (spray, formulation for nebulizer treatment)
viii. Topical (cream, ointment)
ix. Rectum (creams, ointments and suppositories)
x. In the vagina (creams, ointments and pessaries)
xi. Ophthalmology (drops and ointments)
xii. Oral (chewable and now chewable tablets)

Many of the drugs discussed herein, particularly those listed below, exhibit characteristic strong hydrophobicity and are easily present, for example, in contact with a body part (eg, gastric fluid), for example, in the presence of very small amounts of water. Precipitate. Thus, for example, it is extremely difficult to provide an oral formulation that is acceptable to a patient and is stable in storage, stable in storage, suitable for regular administration, and capable of controlled patient administration.

Proposed liquid preparations, for example liquid preparations for oral administration of many drugs shown in the tables of the present specification, have so far assumed that ethanol, oil, or similar excipients have mainly been used as carrier media. As Thus, many commercially available drug drinks use ethanol and olive oil or corn oil as a carrier medium, for example, with ethanol and LAB RIFIL, and solvent systems containing equivalent excipients as the carrier medium. Yes. For example, commercially available cyclosporin drinks are used with surfactant labroids with ethanol and olive oil or corn oil as carrier media. See, for example, U.S. Pat. No. 4,388,307. However, the use of drinks and similar compositions proposed in the art is associated with various difficulties.

Furthermore, it has been clarified that known oil-based systems have problems in palatability. The taste of known drinks of some drugs is particularly unpleasant. Appropriate flavored beverages, such as chocolate beverage preparations mixed at high dilution just prior to consumption, have been commonly used so that they are all acceptable under normal treatment. Employing an oily system also requires the use of high concentrations of ethanol, but the use of ethanol itself is essentially undesirable, especially if it is anticipated for administration to children. In addition, evaporation of ethanol from, for example, capsules (as widely discussed to address palatability issues as discussed) or other forms (eg, open) causes drug precipitation. When such compositions are provided, for example, in the form of encapsulated in soft gelatin, the particular difficulty creates the need for the encapsulated product to be airtight components, such as airtight blisters or aluminum foil blister packaging. . In other words, this makes the product bulky and raises manufacturing costs. Moreover, the storage characteristics of the formulation are far from ideal.

For many of the drugs described herein, the level of bioavailability obtained using existing oral dosing systems is also low, greatly dependent on time between treatments, between individuals, individual patient types, and even in the same individual. Show significant fluctuations. Reports in the literature show that currently available treatments with commercially available drink drugs have an average absolute bioavailability of only about 10-30%, between populations such as the liver (relatively low bioavailability) and There is significant variability between transplanted patient groups of bone marrow (relatively high bioavailability). The reported variation in bioavailability among subjects is between 1 or several percent in some patients but reaches up to 90% or more in other patients. As already mentioned, it is often observed that an individual's bioavailability varies significantly over time. Therefore, there is a need for uniform and high bioavailability to patients for many of the drugs shown herein.

The use of such dosage forms is also characterized by extreme variations in required patient dosing. In order to achieve effective treatment, the blood or serum level of the drug must be maintained within a certain range. In other words, this need is based on the specific condition being treated, such as whether the treatment will block one or more pharmacological effects of a particular drug, and in conjunction with the main treatment. Depending on the case, it will fluctuate greatly. Because the bioavailability obtained with conventional dosage forms varies widely, the daily dose required to obtain the required serum levels also varies greatly between individuals or even within the same individual. This may require periodic and frequent monitoring of blood / serum levels in patients undergoing drug treatment. Blood / serum level monitoring must be performed regularly. This monitoring is necessarily time consuming, inconvenient and greatly increases the overall cost of treatment.

The blood / serum levels of many of the agents described herein, obtained using available dosing systems, may show extreme variation between peaks and valleys. That is, for each patient, the effective drug level in the blood varies greatly between individual medications.

Many of the drugs described herein, particularly beta-lactam antibiotics, cyclosporine, cephalosporin, steroids, quinolone antibiotics and cyclosporine, are also required to be in water-soluble form for injection. Cremaphor L® (CreL), used in current formulations of many drugs, described below, is a polyoxyethylenated derivative of castor oil and is known to be a toxic vehicle. To date, many anaphylactic accidents caused by castor oil components have occurred. At present, there are no preparations for many of these drugs that can be made into an aqueous solution having a concentration required for their low water solubility.

In addition to all these very obvious practical problems, there is the problem that undesired side effects are observed when using available oral dosage forms, as already whispered.

In the art, proposals have been made to address these various issues, including solid and liquid oral dosage forms. However, an unresolved critical problem is that some of the drugs listed below are essentially insoluble in aqueous media, thus allowing convenient use while at the same time effective absorption from the stomach or gastrointestinal tract, for example. And the use of dosage forms that can contain sufficiently high concentrations of the drug to meet the criteria required for bioavailability so that consistently appropriate high blood / serum levels can be achieved.

A particular difficulty facing oral administration of these drugs is that their use in certain drug therapies intended to treat disease states of relatively low severity or risk is necessarily limited . For example, the administration of cyclosporine as a test agent is a particular area of difficulty in this regard, for the treatment of autoimmune diseases and other skin conditions, such as the treatment of atopic dermatitis and psoriasis, and the art. It has been proposed to employ cyclosporine therapy in the treatment of stimulated growth of hair, such as aging and alopecia due to disease, as proposed.

Thus, while oral cyclosporine treatment has shown great potential benefits for the drug, for example, in patients suffering from psoriasis, the risk of side reactions from oral treatment precludes general use. Various proposals have been made in the art for the application of cyclosporine, for example, a description of topical forms of cyclosporine and various topical delivery systems. However, topical application attempts have failed to provide a clear and effective treatment.

However, the present invention overcomes the aforementioned problems. More specifically, the prodrug of the present invention significantly increases the solubility in aqueous solution compared to the non-prodrug form of the drug, thereby providing a carrier such as ethanol or castor oil when administered as a solution. Use can be avoided. In addition, the prodrugs of these drugs according to the present invention do not show the side effects of conventional formulations. Furthermore, many of the drugs in the table below have been found to facilitate oral absorption when administered in the form of a prodrug according to the present invention, thereby significantly increasing its bioavailability and its efficacy.

Preferred agents for use in combination with amino acids that form prodrugs are listed in the table below, and the benefits found are listed in the penultimate column of the table. The important items in the table are as follows:
a) Improved flavor b) Suitable octanol / water partition coefficient (ie water solubility)
c) Improved in vitro and in vivo stability d) Blood brain barrier passage e) Elimination of first-pass effect in liver f) Reduction of enterohepatic recirculation g) Painless injection of parenteral formulation h) Bioavailability Improvement i) Increased absorption rate j) Reduced side effects k) Dose proportionality l) Selective prodrug hydrolysis at the site of action m) Controlled release characteristics n) Targeted drug delivery o) Reduced toxicity, thereby therapeutic ratio Improved p) Reduced dose q) Altered metabolic pathway to deliver more drug to the site of action

In addition, the table also shows the usefulness of prodrugs. The usefulness of a prodrug is the same as the corresponding drug (without the amino acid moiety attached). Usefulness is described in documents such as the Physicians Desk Reference 2004 edition, the contents of which are incorporated by reference.

The following non-limiting examples illustrate the invention in detail.

Synthesis of various amino acid derivatives of selected drugs. The propofol derivative propofol (2,6-diisopropylphenol) is a low molecular weight phenol widely used as a central nervous system anesthetic and has a sedative and hypnotic action. It is administered intravenously during induction and maintenance of mammalian anesthesia and / or sedation. The main advantages of propofol are that it can introduce anesthesia quickly, has minimal side effects, and recovers immediately upon withdrawal without prolonged sedation.

Propofol has been shown to have numerous therapeutic uses, which are extremely diverse and somewhat surprising. For example, it has been shown to be antioxidants, antiemetics, antipruritics, antiepileptics, anti-inflammatory agents, and even thought to have anti-cancer properties.

Mechanism of action:
The mechanism of action of propofol has been extensively studied. Its central nervous system anesthetic action has been shown to be related to its high affinity for specific GABA receptor subclasses (Collins GS, 1988, Br. J. Pharmacology. 542, 225- 232). However, there are various receptors in the brain that are substrates for propofol, and therefore their activities are diverse.

Propofol also has a significant biological effect as an antioxidant. Due to this general activity of propofol, it is theoretically useful for the treatment of many inflammatory processes where oxidation is an important factor. For example, synthesis of prostaglandins via cyclooxygenase causes inflammation. By inhibiting airway oxidation, propofol can be used to treat acid aspiration, adult / childhood dyspnea syndrome, airway obstruction disease, asthma, cancer, and many other similar conditions.

Propofol is associated with Parkinson's disease, Alzheimer's disease, Friedrich's disease, Huntington's disease, multiple sclerosis, amyotrophic lateral sclerosis, spinal cord injury, and various other neurodegenerative diseases because oxidative tissue damage is quite common. It has been said to be useful in the treatment of.

Propofol is currently sold in the US market as an emulsion for intravenous injection under the trade name Diprivan (registered trademark) by Astra Zeneca. This is one of the most widely used commercial short-acting central nervous system anesthetics. The concentration of propofol is 10 mg / mL in a non-pyrogenic sterile emulsion and the formula includes soybean oil, glycerol, egg lecithin, disodium edetate and sodium hydroxide.

A critical drawback of propofol is that it does not dissolve in water at all. Even at very low concentrations of 10 mg / mL, this drug precipitates out of aqueous solution at room temperature. Therefore, the manufacturer of this formulation has taken a bold approach of emulsifying the product in water using extremely complex and harmful emulsifiers. For example, manufacturers of IV formulations use egg lecithin, Cremaphor L®, castor oil, and other similar emulsifiers.

However, the use of such emulsifiers is associated with various problems. Various types of Cremaphor L® emulsifiers are known to cause allergic reactions. Egg lecithin and castor oil are known to cause anaphylactic shock in some patients. Furthermore, it is short-term and more expensive for propofol to remain stable in these emulsions. Furthermore, the presence of egg lecithin and castor oil facilitates the growth of microorganisms in the emulsion. Propofol can be dissolved in water by synthesis with cyclodextrin, but cyclodextrin is not approved by the FDA for intravenous therapy.

To date, there has been no example of producing a safe prodrug of propofol. British Patent Nos. 1,102,011 and 1,160,468 and U.S. Pat. No. 3,389,138 describe phenolic esters of various amino acids, where propofol has a number of side chains. , But produces harmful effects when released into the body.

US Pat. No. 6,451,854 describes many substituted α-aminoacetates of propofol, where propofol and side chains are substituted by a number of different chemical groups. Not all of the N, N-2 substituted glycine esters of propofol have been shown to be non-toxic, or many of the compounds described therein are derivatives of propofol. From this, when the ester is cleaved by the enzyme and then released into the body, many of the released active agents are not propofol, so they do not have toxicity data and are completely novel with unknown therapeutic effects on humans. Is a molecule.

In yet another published article on the water-soluble amino acid ester salt of the anesthetic propofol (Int. J. Pharmaceuticals, 175 [2]: 195-204, 1998) the authors synthesized a number of water-soluble propofol derivatives. ing. However, when these prodrugs are cleaved by esterase enzymes, substituted unnatural amino acids with an unknown toxicity profile are released into the body.

To date, no pharmaceutical formulation is commercially available that can deliver propofol without dangerous side effects. However, the present invention has created a number of water-soluble, non-toxic propofol derivatives that are suitable for delivering propofol into the body, without dangerous side effects, without the need for toxic and expensive additives, solubilizers and emulsifiers. .

Accordingly, in one embodiment, the present invention is directed to propofol prodrugs. A prodrug consists of a carboxyl group of an amino acid esterified with a free hydroxyl group present on the propofol molecule.

または薬学的に許容されるその塩であり、式中、ＡＡはアミノ酸であり、ＡＡのカルボキシル基はプロポフォールのヒドロキシル基と反応する。 More specifically, one embodiment of the present invention is a compound of the formula

Or a pharmaceutically acceptable salt thereof, wherein AA is an amino acid and the carboxyl group of AA reacts with the hydroxyl group of propofol.

In another embodiment, the present invention is directed to a pharmaceutical composition comprising a therapeutically effective amount of the various propofol prodrugs described above and a pharmaceutical carrier thereof.

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

In a further embodiment, the present invention is directed to a method of enhancing the solubility of propofol in an aqueous solution comprising reacting the hydroxyl functionality of propofol and isolating the product.

In yet another embodiment, the present invention significantly reduces the possible adverse side effects of current formulations containing toxic excipients when administered to a patient, significantly and therapeutically, or A method of eliminating reacting the hydroxyl functionality of a propofol molecule with the carboxyl functionality of a selected amino acid to form an ester covalent bond, respectively, and isolating the product, and said production It is directed to a method comprising administering an object to a patient.

The present invention shows that when an unsubstituted natural amino acid is esterified to propofol, the resulting prodrug is highly water soluble (> 200 mg / L to water) and releases non-toxic amino acids when cleaved in the body. And no need for toxic emulsifiers, additives and other excipients.

Furthermore, although the present invention also shows that drugs are produced, they have also been shown to be prodrugs of the propofol of the present invention and highly effective central nervous system anesthetics. Thus, the amino acid prodrug is an effective central nervous system anesthetic with or without the release of the active parent drug.

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

The prodrugs of the present invention are believed to have no antioxidant activity because the phenolic group responsible for antioxidant activity is blocked; however, the present invention is associated with propofol prodrug release of propofol It has been found to be an effective anesthetic with or without. The described propofol prodrug releases propofol when administered in vivo, and the resulting drug retains its pharmacological properties and antioxidant properties.

The prodrugs of the propofol of the present invention clearly provide many advantages over propofol, for example, all of the side chains cleaved from these prodrugs are natural amino acids and are therefore non-toxic. As a result, a high therapeutic index is obtained. Second, all prodrugs are easily cleaved in the body to release propofol. In addition, due to their high water solubility, they make in-situ solutions or provide pre-filled medicinal solutions in syringes or bottles for injection, just prior to IV administration using lyophilized sterile powders Can be easily administered. Amino acid esters are more stable than propofol because the OH group in propofol is blocked against oxidation. Therefore, the propofol prodrugs of the present invention do not have the toxicity and other pharmaceutical problems found in currently marketed formulations and are more effective than profolol itself.

Propofol prodrugs of the present invention have anti-inflammatory, antioxidant, anti-cancer, anti-convulsant, antiemetic and anti-pruritic properties.

These propofol prodrugs of the present invention are effective in the treatment of diseases or conditions in which propofol is commonly used. The prodrugs disclosed herein deform in the body, release the active compound, and reduce or eliminate the biopharmaceutical and pharmacokinetic obstacles associated with each prodrug, thereby propofol therapeutic benefits To increase. However, it should be noted that these prodrugs themselves have sufficient activity in mammals without releasing the active drug. Because the prodrug is more water soluble than propofol, it need not be associated with a carrier vehicle that may be toxic or cause unwanted side effects, such as alcohol or castor oil. Furthermore, oral formulations containing propofol prodrugs are absorbed into the blood and are extremely effective.

Thus, the prodrugs of the present invention enhance the therapeutic benefit by removing the biopharmaceutical and pharmacokinetic obstacles of existing drugs.

Furthermore, prodrugs can be easily synthesized in high yield using commercially available reagents that are readily available.

Overview:
The synthetic sequence of propofol glycine, L-proline, and L-lysine ester is described below. However, these are exemplary and any of the amino acid prodrugs can be prepared using the following methodology. Complete procedures and analytical data are given in the experimental section. In general, propofol (10 g) is prepared in the presence of a catalytic amount of 4- (N, N-dimethylamino) -pyridine (DMAP) as shown in the scheme below. Coupling with N-Boc protected amino acid (1 equivalent) using 3-ethylcarbodiimide hydrochloride (EDC). EDC was removed by extraction with water. After drying over sodium sulfate, filtration and concentration, the propofol crude protected amino acid ester was purified by flash chromatography to yield the protected ester in 50-60% yield. The protected ester was then stirred at room temperature in diethyl ether saturated with hydrochloric acid (gas) to remove the protecting group. The yield of the deprotection step was generally 60 to 95%. After filtration and drying, there was no need for further purification of the propofol glycine and L-proline ester hydrochloride. The hydrochloride salt of L-lysine-propofol ester was crystallized once from ethanol to remove traces of mono-protected L-lysine-propofol ester.

Composition order

Overview Synthesis of glycine, L-proline and L-lysine esters of propofol: a) EDC, DMAP, CH ₂ Cl ₂ : b) HCl (g), Et ₂ O

The synthesis of the experimental chapters SPI0010, SPI0011 and SPI0013 was carried out in batches. In general, small scale experiments were performed first, followed by larger batches. Reagents listed in the experimental section were purchased from Aldrich, Across or Bachem with the highest purity available except for solvents purchased from Fischer Scientific or Mallinkrodt.

ｔ−ブトキシカルボニルアミノ−酢酸２，６−ジイソプロピルフェニルエステル：

^１Ｈ 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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_３） ： δ＝ １６９．３５， １５５．７５， １４５．２２， １４０．３５， １２６．９０， １２４．１４， ８０．３２， ４２．６６， ２８．５４， ２７．７９， ２３．５７． 1) SPI0010
Propofol (9.98 g, 55.97 mmol) was dissolved in dichloromethane (200 ml) at room temperature under an argon atmosphere. N-t-butyloxocarbonyl-glycine (11.2 g, 63.91 mmol) to 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC, 11.1 g, 57.9 mmol) and 4- (N, N-dimethylamino) -pyridine (DMAP, 1.5 g, 12.27 mmol) was added together. After stirring for 21 hours at room temperature under an argon atmosphere, water (200 mL) was added and the layers were separated. The dichloromethane layer was washed once more 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 using silica gel (250 g) eluting with hexanes / ethyl acetate (10: 1). This procedure gave the protected propofol N-BOC protected glycine ester (11.34 g, 60% yield) as a white solid.

t-Butoxycarbonylamino-acetic acid 2,6-diisopropylphenyl 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, CDC1 ₃ ): δ = 169.35, 155.75, 145.22, 140.35, 126.90, 124.14, 80.32, 42.66, 28.54, 27 79, 23.57.

アミノ−酢酸２，６−ジイソプロピル−フェニルエステル・塩酸塩：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_３） ： δ ＝ １６６．４２， １４４．８４， １４０．４２， １２７．１０， １２４．０６， ４０．４７， ２７．６１， ２３．５５．

ＣＨＮ分析：
ｃａｌｃ． ： Ｃ ６１．８７， Ｈ ８．１６， Ｎ ５．１５ ； ｆｏｕｎｄ： Ｃ ６１．１４， Ｈ ８．２０， Ｎ ５．１４．
Propofol-Boc-glycine ester (11.28 g, 33.6 mmol) was dissolved in anhydrous diethyl ether (200 mL) at room temperature. Hydrochloric acid (gas) was passed through the solution with stirring for 45 minutes. The mixture was kept stirring at room temperature for 48 hours under an argon atmosphere. After 48 hours, hexanes (200 mL) were added and the precipitate was filtered. The white solid was dried at 88 ° C. under high vacuum for 5 hours. The experiment yielded SPI0010 (8.73 g, 95% yield, 99.9% purity by HPLC) as a white solid.

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

¹ H NMR (300 MHz, CDC1 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.

ピロリジン−１，２−ジカルボン酸１−ｔｅｒｍ−ブチルエステル２−（２，６−ジイソプロピル−フェニル）エステル：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_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）．

^１３Ｃ 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． 2) SPI0011
Propofol (10.03 g, 56.23 mmol) was dissolved in dichloromethane (100 ml) at room temperature under an argon atmosphere. Nt-butyloxocarbonyl-L-proline (14.04 g, 65.22 mmol) was converted to 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC, 11.95 g, 62.33 mmol). ) And 4- (N, N-dimethylamino) -pyridine (DMAP, 1.1 g, 9.0 mmol). After stirring at room temperature for 3 hours under an argon atmosphere, water (100 mL) was added and the layers were 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 hexanes / ethyl acetate (10: 1). By this procedure, N-BOC-protected L-proline ester of protected propofol (11.34 g, 66% yield) was obtained as a transparent oil that solidified upon storage in the refrigerator.

Pyrrolidine-1,2-dicarboxylic acid 1-term-butyl ester 2- (2,6-diisopropyl-phenyl) ester:

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 7.31-7.20 (m, 3H), 4.73 (m, 1H), 3.70-3.50 (m, 2H), 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, CDC 1 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.

ピロリジン−２（Ｓ）−カルボン酸−２，６−ジイソプロピル−フェニルエステル・塩酸塩：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_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）．

^１３Ｃ 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分析：
ｃａｌｅ． ： Ｃ ６５．４８， Ｈ ８．４０， Ｎ ４．４９ ； ｆｏｕｎｄ： Ｃ ６５．５０， Ｈ ８．４３， Ｎ ４．５０． Propofol-Boc-L-proline ester (13.95 g, 37.14 mmol) was dissolved in anhydrous diethyl ether (100 mL) at room temperature. Hydrochloric acid (gas) was passed through the solution with stirring for 60 minutes. The mixture was kept stirring at room temperature for 22 hours under an argon atmosphere. After 22 hours, hexanes (50 mL) were added and the precipitate was filtered. The white solid was dried at 88 ° C. under high vacuum for 5 hours. From the experiment, SPI0011 (9.1 g, yield 81%, purity 99.1% by HPLC) was obtained as a white solid.

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

¹ H NMR (300 MHz, CDC1 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:
call. : C 65.48, H 8.40, N 4.49; found: C 65.50, H 8.43, N 4.50.

3) SPI0013
Di-N-boc-L-lysine dicyclohexylamine salt (23.62 g, 0.0447 mmol) was cooled in an ice / water bath with diethyl ether (200 mL) and sodium hydrogen sulfate (9.14 g) in water (200 mL). ). After stirring for 20 minutes, the layers were separated. The ether layer was extracted 3 times with cold water (100 mL). The ether layer was then dried over sodium sulfate (15 g) for 1 hour, filtered and concentrated under reduced pressure. This procedure yielded the free acid of N, N′-di-boc-L-lysine (15.5 g, 100% recovery).

２（Ｓ），６−ビス−ｔ−ブトキシカルボニルアミノ−ヘキサン酸２，６−ジイソプロピル−フェニルエステル

^１Ｈ 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， １２Ｈ）．

^１３Ｃ 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 (8.0 g, 45 mmol) was dissolved in dichloromethane (100 ml) at room temperature under an argon atmosphere. N, N′-di-t-butyloxocarbonyl-L-lysine (15.5 g, 44.7 mmol) was converted to 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC, 8.62 g). 45 mmol) and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.55 g, 4.5 mmol). After stirring at room temperature for 3 hours under an argon atmosphere, water (100 mL) was added and the layers were 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 hexanes / ethyl acetate (9: 1). This procedure gave the protected propofol N-BOC protected L-lysine ester (12.42 g, 55% yield) as a white foam.

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, 12H).

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.

２（Ｓ），６−ジアミノ−ヘキサン酸２，６−ジイソプロピル−フェニルエステル・二塩酸塩；

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣｌ_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分析：
ｃａｌｃ． ： Ｃ ５６．９９， Ｈ ８．５０， Ｎ ７．３８ ； ｆｏｕｎｄ： Ｃ ５６．４８， Ｈ ８．５６， Ｎ ７．３０． Propofol-di-Boc-L-lysine ester (12.34 g, 24.37 mmol) was dissolved in anhydrous diethyl ether (250 mL) at room temperature. Hydrochloric acid (gas) was passed through the solution for 60 minutes with stirring and cooling in an ice / water bath. The mixture was kept stirring at room temperature for 48 hours under an argon atmosphere. After 48 hours, the precipitate was filtered and crystallized from ethanol (100 mL). The white solid was dried at 90 ° C. under high vacuum for 4 hours. From the experiment, SPI0013 (5.5 g, yield 60%, purity by HPLC 98.6%) was obtained as a white solid.

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

¹ H NMR (300 MHz, CDC1 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, CDCl 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. Non-steroidal anti-inflammatory drugs (NSAIDs) prodrugs NSAIDs include a group with a characteristic structure in which a carboxylic acid moiety is added to an aromatic functional surface, for example: acetylsalicylic acid, salicylic acid, diflunisal , Ibuprofen, fenoprofen, carprofen, flubiprofen, ketoprofen, naproxen, sulindac, indomethacin, etodolac, tolmethine, ketorolac, diclofenac and meclofenamate. NSADI has anti-inflammatory, analgesic, antipyretic and anticoagulant activities.

ＮＳＡＩＤは、急性および慢性の疼痛の治療、浮腫、炎症性関節疾患から生じた組織損傷の管理等に広く用いられており、心筋梗塞の治療に於いては効果的な抗凝固剤である。多くの薬剤は、鎮静および抗炎症作用に加えて解熱活性も有しており、熱を下げるのに有効である。 An example of the chemical structure of this unique compound class that exhibits a wide range of pharmacological activities is shown below.

NSAIDs are widely used for the treatment of acute and chronic pain, the management of tissue damage resulting from edema, inflammatory joint diseases, and the like, and are effective anticoagulants in the treatment of myocardial infarction. Many drugs have antipyretic activity in addition to sedative and anti-inflammatory effects and are effective in lowering fever.

Several drugs from the above group are also prescribed for rheumatoid arthritis, osteoarthritis, acute gout, ankylosing spondylitis, and dysmenorrhea.

Mechanism of action:
The primary mechanism by which NSAIDs produce their therapeutic effect is through inhibition of prostaglandin synthesis. Specifically, NSAIDs inhibit cyclooxygenases such as COX-1 and COX-2 enzymes, but these two types of enzymes are responsible for the synthesis of prostaglandins. The OCX-1 enzyme is important for the regulation of platelet aggregation, the regulation of renal and gastric blood flow, and the regulation of gastric acid secretion, whereas the COX-2 enzyme plays an important role in pain and inflammatory processes . NSAIDs significantly extend clotting time and can be used to prevent thromboembolism and myocardial infarction.

All NSAIDs are relatively medium to strong organic acids with a pKa of 3-6. Most of them are carboxylic acid derivatives. Acidic groups are essential for COX inhibitory activity and at physiological pH all NSAIDs are ionized. They all have a very diverse hydrophilic-lipophilic balance and they function by taking their aryl, aromatic and aliphatic side chains, and various other heterocyclic structures. Most NSAIDs bind strongly to plasma proteins and often competitively replace other drugs with similar affinity for plasma proteins. Therefore, co-administration of NSAIDs with other therapeutic categories must be carefully exercised so that there is no drug interaction. Most drugs are metabolized by conjugation in mammals due to acidic carboxyl groups. The major pathway of metabolic clearance of many NSAIDs is glucuronidation followed by elimination from the kidney.

The use of acetylsalicylic acid (aspirin) to prevent coronary heart disease is well known today and has been shown to have saved many myocardial infarction patients. Several other uses of aspirin have been reported, for example, in the medical journal Lancet (Vol 349, p1641), in patients with aspirin showing early signs of a transient ischemic heart attack It has been reported to reduce risk. Preeclampsia and fetal dysgenesis, both due to placental vascular blockage, are the most common complications of pregnancy-preeclampsia occurs millions of cases worldwide each year. In a study involving more than 9000 women in 16 countries, a daily dose of 60 mg aspirin reduced the risk of pre-eclampsia by 13 percent (Aspirin Foundation website). Aspirin has also been shown in several studies to be effective in preventing colorectal, lung, and pancreatic cancer in postmenopausal women. Since aspirin can improve blood flow, it is becoming clear that it is useful for treating certain types of dementia such as diabetes and Alzheimer's disease.

Due to their unique pharmaceutical potential, NSAIDs have received much attention in the press. The main clinical research areas related to the above drugs are related to nonsteroidal anti-inflammatory drugs, especially pain, arthritis (rheumatic and bone), other inflammatory reactions, application to patients with fever, prevention of coronary heart disease. These drugs are also used to treat migraine, menstrual syndrome, back pain, and gout.

NSAIDs offer a great contribution, but more effective and convenient means of administration (e.g. herbal formulations, e.g. oral dosage forms, they are both convenient and at the same time provide appropriate bioavailability for the patient, Have been reported to cause undesirable side reactions; specifically, severe gastroduodenal ulcers, Mucosal erythema and edema, erosion, perforation, bloody stool, ulcerative colitis are major obstacles to their wider use or application. Double injury theory involves direct damage mediated by NSAIDs, followed by systemic effects in which prostaglandin synthesis is inhibited. Local injury may occur as a result of the excretion of active liver metabolites in the bile and subsequent gastroduodenal reflux (Arthritis and Rhematism 1995; 38 (1): 5-18). The effect is additive; either local or systemic mechanisms are sufficient to damage the gastroduodenal mucosa.

Furthermore, the NSAID is very hydrophobic as a characteristic, and only in the presence of a very small amount of water, for example, forms a precipitate easily only by contact with the body (for example, gastric juice). Thus, for example, to provide an oral formulation that can be accepted by the patient, for example in terms of form and taste, can be stably stored, can be administered regularly, can be properly administered to the patient, and can be controlled Is extremely difficult.

Therefore, the proposed liquid preparations, for example liquid preparations for oral administration of NSAID, are basically based on the use of natural gums such as xanthan, cellulose, citric acid and lime flavor. See, for example, US Pat. No. 5,780,046. Commercially available NSAID drinks use incompatible orange and berry flavors, citric acid, xanthan gum, polysorbate 80, pregelatinized starch, glycerin, sodium benzoate, and other artificial colors and flavors. However, the use of drink liquids and similar compositions as proposed in the art is associated with various difficulties.

Furthermore, known oil-based systems have been found to have problems with palatability. The taste of the known drink liquid is particularly unpopular. In normal treatment, it is generally mixed with a beverage with a suitable flavor just before ingestion, such as a chocolate beverage, at a high dilution rate in order to be accepted. Employing an oil-based system also requires the use of higher concentrations of ethanol, and alcohol itself is not desirable, especially if it is anticipated for administration to children. In addition, evaporation of ethanol from, for example, capsules (as widely discussed to address palatability issues, as discussed) or other shapes (eg, open) results in drug precipitation. When such products are encapsulated in, for example, soft gelatin, it is particularly difficult to package the encapsulated products in hermetic components, such as hermetic blister or aluminum foil blister packaging. This makes the product bulky and increases manufacturing costs. Moreover, the storage characteristics of the formulation are far from ideal.

NSAID irritation to the stomach is a matter of great interest not only to the attending physician but also to the patient. Aspirin, fenoprofen, flurbiprofen, indomethacin, ketorolac, meclofenamate, mefenamic acid (and the rapid use of piroxicam cause serious GI side effects. Even ibuprofen is severe when used for long periods of time. Gastrointestinal toxicity is the most frequently experienced side effect associated with NSAIDs and is a major concern.Almost half of all hospitalized cases with hemorrhagic ulcers NSAIDs during the week before hospitalization, use of aspirin, or combined use (Faulkner G, Prichard P, Somerville K, et al. Aspirin and bleeding petit cercers in the der. 297: 1311-1313) According to a survey of Tennessee Medicaid patients hospitalized for GI complications, patients using NSAIDs develop GI bleeding or peptic ulcer disease compared to patients who did not take NSAIDs The risk is four times higher (Griffin MR, Piper JM, Daugherty JR, et al. Nonsteroidal anti-inflammatory drug use and increaked risk for peptic ulcer. According to 2% to 4% of patients receiving continuous NASID treatment for rheumatoid arthritis, severe GI injury every year Gastric ulcers (4.725), duodenal ulcers (1.1 to 1.6), bleeding (3.8) when compared with patients not using NSAIDs and patients using NSAIDs The relative risk of perforation, and death all increase with the use of NSAIDs.In 1989, the total number of hospitalizations for patients with rheumatoid arthritis is approximately 20,000 hospitalizations / year, with an estimated cost per hospitalization of $ 10 (Fries JF, Miller SR, Spitz PW, et al. Howard an epidemiology of gastropathy associated with non-inflammatory energy. J Gastroenterology. 1989; 96: 647-655).

There is also a need to provide NSAIDs in water-soluble form for injection. It is well known that the high concentrations of alcohol and tromethamine used to form the salts of current catorolac formulations are harmful. At present, there is no preparation that can make NSAID into an aqueous solution having a necessary concentration because the drug has low water solubility.

It is clearly much more difficult in practice to have the undesirable side effects observed when using available oral dosage forms, as already mentioned, than all of these.

Several proposals have been made in the art to address these various problems, including both solid and liquid oral dosage forms. However, NSAIDs are still inherently insoluble in aqueous media, and therefore can be conveniently used, and can be effectively absorbed, for example, from the stomach and stomach cavity, and are stable and of appropriate height. Preventing the use of dosage forms that can contain sufficient concentrations of NSAIDs that meet the required criteria in terms of bioavailability / to achieve serum levels remains a challenge to overcome.

This prodrug of NSAID overcomes the above problems. More specifically, embodiments of the present invention provide a NSAID prodrug that greatly enhances its solubility in aqueous solution, thereby eliminating the need to use a carrier such as ethanol or castor oil as a solution. It is aimed. Furthermore, the NSAID prodrugs according to the present invention do not show the side effects of the prior art formulations. Furthermore, the prodrugs of the present invention have almost no stomach irritation upon oral administration, thereby greatly increasing the therapeutic index of the prodrugs tested and their efficacy.

または、薬学的に許容されるその塩を有する；式中ＹはＮＨ−ＡＡまたはＯ−ＡＡであり、ＡＡはアミノ酸であって、ＡＡのアミノ基またはヒドロキシル基がＮＳＡＩＤのカルボキシル酸基と反応する。 Accordingly, in one aspect, the present invention is directed to prodrugs of NSAIDs. Preferred prodrugs of NSAID have the following formula:

Or having a pharmaceutically acceptable salt thereof, wherein Y is NH-AA or O-AA, AA is an amino acid, and the amino group or hydroxyl group of AA reacts with the carboxylic acid group of NSAID .

The present invention is also directed to pharmaceutical compositions comprising a therapeutically effective amount of the various NSAIDs described above and pharmaceutically acceptable carriers therefor.

In another embodiment, the present invention is directed to a method of treating a patient in need of NSAID treatment comprising administering to said patient an effective amount of NSAID.

In a further embodiment, the present invention is directed to a method for increasing the solubility of an NSAID in an aqueous solution comprising reacting the carboxyl functionality of each NSAID and isolating the product.

In yet another embodiment, the present invention comprises reacting the carboxyl functionality of each NSAID molecule with the amine or hydroxyl function of a selected amino acid to form an amide or ester covalent bond, respectively, and the product simply. It is aimed at a method of reducing or eliminating gastric mucosal damage due to NSAIDs upon patient administration in a substantially and therapeutically effective manner, including releasing and administering the product to the patient.

A. Synthetic overview of ibuprofen amino acid derivatives:
Methods for the synthesis of L-serine, L-threonine and L-hydroxyproline esters of ibuprofen are outlined in the synthetic order section. Complete operating procedures and analytical data are given in the experimental section. Again, these synthesis summaries are exemplary. This summary can be applied to other amino acids in the preparation of the NSAID prodrugs of the present invention. In general, (±) -ibuprofen (4-10 g, batch) is prepared in the presence of a catalytic amount of 4- (N, N-dimethylamino) -pyridine (DMAP) in the presence of 1- (3-dimethylaminopropyl) -3- Ethylcarbodiimide hydrochloride (EDC, 1 eq) was used to couple with the N-benzyloxy / benzyl ester protected amino acid (1 eq). When the reaction was complete, excess EDC was removed by extraction with water, DMAP was removed by extraction with dilute acid, and ibuprofen was removed by extraction with sodium bicarbonate. After drying over sodium sulfate, filtration and concentration, the crude protected amino acid ester of (±) -ibuprofen is used as is or purified by flash chromatography on silica gel to give good yield (85-95% ) Produced a protected ester. If a slight excess of ibuprofen and coupling agent was used, column chromatography was generally not necessary and total extraction was performed. The protecting group was removed by hydrogenation (25-35 psiH ₂ ) in the presence of 10% palladium-carbon and hydrochloric acid. The yield of the deprotection step was 70 to 90%. After filtration and drying, the hydrochloride salt of the serine and threonine ester of (±) -ibuprofen was purified by crystallization. The hydrochloride salt of L-hydroxyproline-ibuprofen ester was a gel and did not solidify / crystallize. In this case, hydrogenation was repeated without using an acid to purify the neutral compound.

Since ibuprofen starts as a mixture of enantiomers, the final product is produced as a mixture of diastereomers, except for the threonine ester. In the case of the threonine ester of ibuprofen, the final diastereomeric salt can be easily separated by washing with water, acetone or acetonitrile. The insoluble isomer (SPI0016A) was determined to be the active isomer by comparison with an authentic standard prepared from S-(+)-ibuprofen. Serine and hydroxyproline esters of (±) -ibuprofen could not be easily separated by this method.

Synthesis order:

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

Experiment chapter:
The synthesis of SPI0015, SPI0016 and SPI0017 was performed in 2 or 3 batches. The reagents listed in the experimental section were purchased with the highest purity available from Sigma-Aldrich, Acros or Bachem, except for solvents purchased from Fischer Scientific or Mallinkrodt.

２（Ｓ）−ベンジルオキシカルボニルアミノ−［２（Ｒ，Ｓ）−（４−イソブチル−フェニル）−プロピオニルオキシ］−プロピオン酸ベンジルエステル：

^１Ｈ 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）．

^１３Ｃ 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， １２７．１９， １２７．１６， ６７．７５， ６７．３２， ６４．５１， ６４．３２， ５３．７１， ４５．１６， ４５．０２， ３０．３５， ２２．６０， １８．２７． 1) Preparation of (±) -ibuprofen-L-serine ester hydrochloride (SPI10015) (±) -ibuprofen (5.04 g, 24.4 mmol), N-carbobenzyloxy-L-serine benzyl ester (8. 11 g, 24.6 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC, 4.87 g, 25.4 mmol) and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.40 g, 3.27 mmol) was dissolved in dichloromethane (150 ml) at room temperature under argon atmosphere. After stirring at room temperature for 22 hours under an argon atmosphere, water (100 mL) was added and the layers were separated. The dichloromethane layer was washed once more 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 using silica gel (250 g) eluting with hexanes / ethyl acetate (3: 1). This procedure afforded protected L-serine- (±) -ibuprofen ester SPI001501 (11.4 g, 90% yield) as a colorless solid.

2 (S) -Benzyloxycarbonylamino- [2 (R, S)-(4-isobutyl-phenyl) -propionyloxy] -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, 127.19, 127.16, 67.75, 67.32, 64.51 64.32, 53.71, 45.16, 45.02, 30.35, 22.60, 18.27.

２（Ｓ）−アミノ−３−［２（Ｒ，Ｓ）−（４−イソブチルフェニル）−プロピオニルオキシ］−プロピオン酸・塩酸塩；（（Ｒ，Ｓ）−イブプロフェン−Ｌ−セリンエステル・塩酸塩）：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＤＭＳＯ）： δ ＝ ８．９２ （ｂｒ ｓ， ３Ｈ）， 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）．

^１３Ｃ 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分析 ：
９９．１３％ ｐｕｒｉｔｙ； ｒｔ ＝ ３．１３３ ｍｉｎ ； Ｌｕｎａ Ｃ１８ ５ｕ ｃｏｌｕｍｎ （ｓｎ １６７９１７−１３）； ４．６ｘ２５０ ｍｍ； ２５４ ｎｍ ； ５０％ ＡＣＮ／５０％ ＴＦＡ ｂｕｆｆｅｒ （０．１％） ； ３５ Ｃ； ２０ ｕｌ ｉｎｊ．； １ ｍｌ／ｍｉｎ ； １ ｍｇ／ｍＬ ｓａｍｐｌｅ ｓｉｚｅ； ｓａｍｐｌｅ ｄｉｓｓｏｌｖｅｄ ｉｎ ｍｏｂｉｌｅ ｐｈａｓｅ．

ＣＨＮ分析：
ｃａｌｃ． ： Ｃ ５８．２７， Ｈ ７．３３， Ｎ ４．２５ ； ｆｏｕｎｄ： Ｃ ５８．４４， Ｈ ７．４６， Ｎ ４．２５．

融点： １６９．５−１７０．５ ℃ Protected ibuprofen-L-serine ester (22.50 g, 43.4 mmol) is dissolved in ethanol (200 mL) at room temperature and charged to a Parr bottle containing 10% palladium-carbon (3.86 g, 50% wet rate) with nitrogen. Added in the atmosphere. Hydrochloric acid (10 mL of 37% HCl in 30 mL of water) was added and the nitrogen atmosphere was replaced with hydrogen gas (25 psi). After shaking for 4 hours, the palladium catalyst was removed by filtration through Celite. Ethanol / water was removed under reduced pressure. The remaining white solid was washed with water (25 mL), acetone (20 mL) and dried under high vacuum (4 hours, 88 ° C.). From this experiment, (±) -ibuprofen-L-serine ester / hydrochloride SPI0015 (11.3 g, yield 80%) was obtained as a colorless solid.

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

¹ H NMR (300 MHz, DMSO): δ = 8.92 (br s, 3H), 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 × 250 mm; 254 nm; 50% ACN / 50% TFA buffer (0.1%); 35 C; 20 ul inj. 1 ml / 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 ° C

２（Ｓ）−ベンジルオキシカルボニルアミノ−３−［２（Ｒ，Ｓ）−（４−イソブチル−フェニル）−プロピオニルオキシ］−酪酸ベンジルエステル：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_３） ： δ ＝ 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， ｌ／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， １．５ Ｈ， Ｊ＝ ６．６ Ｈｚ）， １．１７ （ｄ， １．５ Ｈ， Ｊ＝ ６．６ Ｈｚ）， ０． ８５ （ｍ， ６ Ｈ）．
^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_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， ７０．９７， ７０．７０， ６７．８１， ６７．６６， ６７．５３， ５７．８３， ４５．１９， ３０．３９， ２２．６１， １８．５７， １８．３０， １７．１８， １６．８７． 2a) Preparation and separation of (±) -isoprofen-L-threonine ester hydrochloride (SPI0016A and SPI0016B) (±) -isoprofen (4.15 g, 20.11 mmol) N-carbobenzyloxy-L-threonine Benzyl ester (6.90 g, 20.11 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC, 3.95 g, 20.6 mmol) and 4- (N, N- Dimethylamino) -pyridine (DMAP, 0.25 g, 2.0 mmol) was dissolved in dichloromethane (50 mL) at room temperature under an argon atmosphere. After stirring for 19 hours, the dichloromethane layer was washed with water (50 mL), 5% hydrochloric acid (2 × 25 mL), water (25 mL), saturated sodium bicarbonate (2 × 25 mL) and water (50 mL). After drying over sodium sulfate (5 g) for 1 hour, filtration, and concentration under reduced pressure, the remaining oil was used without further purification. This procedure gave the protected L-threonine- (±) -ibuprofen ester SPI001601 (10.2 g, 95.3% yield) as a pale yellow oil, which was allowed to solidify.

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

¹ H NMR (300 MHz, CDC 1 ₃ ): δ = 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, l / 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, 1.5 H, J = 6.6 Hz), 1.17 (d, 1.5 H, J = 6.6 Hz), 0. 85 (m, 6 H).
¹³ C NMR (75 MHz, CDC 1 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, 70.97, 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 mmol) is dissolved in warm ethanol (150 mL) and charged to a Parr bottle containing 10% palladium-carbon (3.4 g, 50% wetness) with nitrogen. Added in the atmosphere. Hydrochloric acid (6 mL of 37% HCl in 20 mL of water) was added and the nitrogen atmosphere was replaced with hydrogen gas (30 psi). After shaking for 3 hours, the palladium catalyst was removed by filtration through Celite (30 g). Ethanol / water was removed under reduced pressure. From this experiment, (±) -ibuprofen-L-threonine ester / hydrochloride (SPI0016A and SPI0016B, 6.4 g, crude raw material yield 95%) was obtained as a colorless solid. The crude mixture of diastereomers was stirred in acetone (200 mL) for 2 hours at room temperature under an argon atmosphere. After 2 hours, the solid (2.84 g, SPI0016A) was filtered. The filtrate (SPI0016B, 3.0 g) was concentrated under reduced pressure.

２（Ｓ）−アミノ−３（Ｒ）−［２（Ｓ）−（４−イソブチル−フェニル）−プロピオニルオキシ］−酪酸；（Ｓ−イブプロフェン−Ｌ−スレオニンエステル・塩酸塩，活性異性体）：

^１Ｈ 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）．

^１３Ｃ 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分析：
ｃａｌｃ．： Ｃ ５９．３８， Ｈ ７．６２， Ｎ ４．０７ ； ｆｏｕｎｄ： Ｃ ５９．１７， Ｈ ７．６３， Ｎ ４．０４．

ＨＰＬＣ分析：
９８．２８％ ｐｕｒｉｔｙ； ｒ． ｔ． ＝ ６．９５１ ｍｉｎ．； ６０％ ＴＦＡ （０． １％）／４０％ ａｃｅｔｏｎｉｔｒｉｌｅ； １ ｍＬ／ｍｉｎ； ３７．５ Ｃ； Ｌｕｎａ Ｃ１８， ３ｕ ｃｏｌｕｍｎ （ＳＮ １６７９１７−１３）， ４．６ｘ２５０ ｍｍ； ２２ ｕｌ ｉｎｊｅｃｔｉｏｎ．

旋光度： ＋ ２４．５ °

融点： １８９−１９０ ℃ 1) Purification of SPI0016A (active isomer) After three batches of S-ibuprofen-L-threonine ester (SPI0016A) were completed, the batches were combined (total 8.78 g), 3 times from DIUF water (100 mL) Crystallized. A small amount of zwitterion was generated each time. To regenerate the salt, the resulting solid (at each crystallization) was dissolved in 1% hydrochloric acid ethanol (3% 37% hydrochloric acid in 100 mL ethanol). The ethanol solution was then concentrated at room temperature under reduced pressure. After the third crystallization and regeneration procedure, the salt (5.6 g) was stirred in acetonitrile (100 mL) for 44 hours at room temperature under an argon atmosphere. The salt was then filtered and dried at 50-55 ° under high vacuum until the weight was constant (5.5 g).

2 (S) -amino-3 (R)-[2 (S)-(4-isobutyl-phenyl) -propionyloxy] -butyric 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; r. t. = 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 ° C

２（Ｓ）−アミノ−３（Ｒ）−［２（Ｒ）−（４−イソブチル−フェニル）−プロピオニルオキシ］−酪酸；（Ｒ−イブプロフェン−Ｌ−スレオニンエステル・塩酸塩・活性異性体）：

^１Ｈ 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）．

^１３Ｃ 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分析：
ｃａｌｃ． ： Ｃ ５９．３８， Ｈ ７．６２， Ｎ ４．０７ ； ｆｏｕｎｄ： Ｃ ５９．３０， Ｈ ７．６０， Ｎ ４．０５．

ＨＰＬＣ分析：
９８．４３％ ｐｕｒｉｔｙ ； ｒ． ｔ． ＝ ６．１９ ｍｉｎ．； ６０％ ＴＦＡ （０．１％）／４０％ ａｃｅｔｏｎｉｔｒｉｌｅ； １ ｍＬ／ｍｉｎ； ３７．５ Ｃ； Ｌｕｎａ Ｃ１８， ３ｕ ｃｏｌｕｍｎ （ＳＮ １６７９１７−１３）， ４．６ｘ２５０ ｍｍ； ２２ ｕｌ ｉｎｊｅｃｔｉｏｎ．

旋光度： ＋ １０．４ °

融点： １７６−１７７ ℃ 2) Purification of SPI0016B (inactive isomer):
After three batches of S-ibuprofen-L-threonine ester (SPI0016B) were completed, the batches were combined (9.02 g total) and crystallized three times from DIUF water (50 mL). A small amount of zwitterion was generated during crystallization. In order to regenerate the salt, the resulting solid was dissolved in 1% ethanolic ethanol (37 mL of 37% hydrochloric acid in 100 mL of ethanol). The ethanol solution was then concentrated at room temperature under reduced pressure. The remaining salt (5.93 g) was crystallized a third time from hot toluene (100 mL) with the addition of a small amount of acetone (1 mL). The salt was then filtered and dried at room temperature under high vacuum until the weight was constant (5.1 g).

2 (S) -amino-3 (R)-[2 (R)-(4-isobutyl-phenyl) -propionyloxy] -butyric acid; (R-ibuprofen-L-threonine ester / hydrochloride / active isomer):

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; r. t. = 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

２（Ｓ）−ベンジルオキシカルボニルアミノ−３−［２（Ｒ，Ｓ）−（４−イソブチル−フェニル）−プロピオニルオキシ］−酪酸ベンジルエステル：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_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， ３ Ｈ， Ｊ＝ ６．３ Ｈｚ）， ０．８６ （ｄ， ６ Ｈ， Ｊ＝ ６．６ Ｈｚ）．

^１３Ｃ 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． 2b) Preparation of S-(+)-Ibuprofen-L-threonine ester hydrochloride standard (SPI0016S) S-(+)-Isoprofen (2.0 g, 9.69 mmol), N-carbobenzyloxy-L Threonine benzyl ester (3.25 g, 9.91 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC, 1.90 g, 9.91 mmol) and 4- (N, N-dimethylamino) -pyridine (DMAP, 0.12 g, 1.0 mmol) was dissolved in dichloromethane (25 mL) at room temperature under an argon atmosphere. After stirring for 4 hours, the dichloromethane layer was washed with water (25 mL), 5% hydrochloric acid (25 mL), saturated sodium bicarbonate (2 × 25 mL) and water (25 mL). After drying over sodium sulfate (5 g) for 1 hour, filtration, and concentration under reduced pressure, the remaining oil was used without further purification. This procedure gave the protected S-(+)-ibuprofen-L-threonine ester SPI001601S (5.01 g, 98% yield) as a pale yellow oil, which was allowed to solidify.

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

¹ H NMR (300 MHz, CDC 1 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, 3 H, J = 6.3 Hz), 0.86 (d, 6 H, J = 6.6 Hz).

¹³ C NMR (75 MHz, CDC 1 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.

２（Ｓ）−アミノ−３（Ｒ）−［２（Ｓ）−（４−イソブチル−フェニル）−プロピオニルオキシ］−酪酸；（Ｓ−イブプロフェン−Ｌ−スレオニンエステル・塩酸塩・活性異性体）：

^１Ｈ 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）．

^１３Ｃ 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分析：
９８．２８％ ｐｕｒｉｔｙ； ｒ． ｔ． ＝ ６．９５１ ｍｉｎ．； ６０％ ＴＦＡ （０．１％）／４０％ ａｃｅｔｏｎｉｔｒｉｌｅ； １ ｍＬ／ｍｉｎ； ３７．５ Ｃ； Ｌｕｎａ Ｃ１８， ３ｕ ｃｏｌｕｍｎ （ＳＮ １６７９１７−１３）， ４．６ｘ２５０ ｍｍ； ２２ ｕｌ ｉｎｊｅｃｔｉｏｎ．

旋光度： ＋ ２６．５ °

融点： １８９−１９０ ℃ Protected S-(+)-ibuprofen-L-threonine ester (5.0 g, 9.40 mmol) was dissolved in warm ethanol (100 mL) and 10% palladium-carbon (1.0 g, 50% wetness) was added. To the containing Parr bottle was added in a nitrogen atmosphere. Hydrochloric acid (1 mL of 37% HCl in 10 mL of water) was added and the nitrogen atmosphere was replaced with hydrogen gas (32 psi). After shaking for 2 hours, the palladium catalyst was removed by filtration through Celite (30 g). Ethanol / water was removed under reduced pressure. From this experiment, S-(+)-ibuprofen-L-threonine ester / hydrochloride SPI0016S (2.8 g, crude raw material yield 85%) was obtained as a colorless solid. The salt was stirred in acetone (50 mL) for 3 hours at room temperature under an argon atmosphere. After 3 hours, the solid (2.24 g, 69% yield) was filtered and dried at high vacuum at room temperature until the weight was constant.

2 (S) -amino-3 (R)-[2 (S)-(4-isobutyl-phenyl) -propionyloxy] -butyric 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; r. t. = 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: + 26.5 °

Melting point: 189-190 ° C

４（Ｒ）−［２−（４−イソブチル−フェニル）−プロピオニルオキシ］−ピロリジン−２（Ｓ）−カルボン酸；（（Ｒ，Ｓ）−イブプロフェン−Ｌ−ヒドロキシプロリンエステル）：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_３） ： 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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_３） ： δ ＝ 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． 3) Preparation of (±) ibuprofen-L-hydroxyproline ester (SPI0017) (±) -ibuprofen (5.10 g, 24.7 mmol), N-carbobenzyloxy-L-hydroxyproline benzyl ester (8.80 g, 24.7 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC, 5.10 g, 26.0 mmol) and 4- (N, N-dimethylamino) -pyridine (DMAP) 0.30 g, 2.40 mmol) was dissolved in dichloromethane (100 mL) at room temperature under an argon atmosphere. After stirring for 24 hours, water (100 mL) was added at room temperature under an argon atmosphere, and the layers were separated. The dichloromethane layer was washed with water (100 mL), 5% saturated sodium bicarbonate (2 × 50 mL) and dried over sodium sulfate (5 g) for 1 hour. After filtration and concentration under reduced pressure, the remaining oil was used without further purification. From this procedure, protected (±) -ibuprofen-L-hydroxyproline ester SPI001701 (11.5 g, 85% yield) was obtained as a 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, CDC1 ₃ ): 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, CDC1 ₃ ): δ = 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.

４（Ｒ）−［２−（４−イソブチル−フェニル）−プロピオニルオキシ］−ピロリジン−２（Ｓ）−カルボン酸；（（Ｒ，Ｓ）−イソプロフェン−Ｌ−ヒドロキシプロリンエステル）：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_３） ： δ ＝ 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）． （ジアステレオマー混合物）

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_３） ： 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， ３４．３１， ３４．１６， ２９．７７， ２２．３４， １８．４３， １８．２３． （ジアステレオマー混合物）

ＨＰＬＣ分析：
１００％ ｐｕｒｉｔｙ； ｒ． ｔ． ＝ 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分析：
ｃａｌｃ．： Ｃ ６７．６９， Ｈ ７．８９， Ｎ ４．３９ ； ｆｏｕｎｄ： Ｃ ６７．４７， Ｈ ７．８７， Ｎ ４．３０．

融点： １９８−１９９ ℃ Protected ibuprofen-L-hydroxyproline ester (11.40 g, 43.3 mmol) is dissolved in ethanol (150 mL) at room temperature and added to a Parr bottle containing 10% palladium-carbon (2.73 g, 50% wetness). In a nitrogen atmosphere. The nitrogen atmosphere was replaced with hydrogen gas (34 psi). After shaking for 5 hours, the palladium catalyst was removed by filtration through Celite. Ethanol was removed under reduced pressure. The remaining white solid (6.60 g) was washed with DIUF water (50 mL), diethyl ether (50 mL) and dried under high vacuum until the weight was constant. From this experiment, protected (±) -ibuprofen-L-hydroxyproline ester SPI0017 (5.64 g, 84% yield) was obtained as a colorless solid.

4 (R)-[2- (4-Isobutyl-phenyl) -propionyloxy] -pyrrolidine-2 (S) -carboxylic acid; ((R, S) -isoprofen-L-hydroxyproline ester):

¹ H NMR (300 MHz, CDC1 ₃ ): δ = 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). (Diastereomer mixture)

¹³ C NMR (75 MHz, CDC1 ₃ ): 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, 34.31, 34.16, 29.77, 22.34, 18.43, 18.23 . (Diastereomer mixture)

HPLC analysis:
100% purity; r. t. = 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 ° C

Efficacy of the synthesis of (±) -ibuprofen L-serine, L-threonine and L-hydroxyproline ester by acetylcholine-induced abdominal contraction in male albino mice (anti-nociceptive):

This test was conducted to evaluate the efficacy of (±) -ibuprofen L-serine, L-threonine and L-hydroxyproline ester as an index in albino mice, considering acetylcholine-induced antagonistic torsion. It was.

Various novel formulations of ibuprofen and reference controls, ibuprofen (racemic mixture) and ibuprofen (S)-(+), 5% (v / v) Tween 80 milliQ water in the vehicle, gavage Were administered to male albino mice (Swiss strain). The study was conducted with the vehicle control group at two dosage levels, 50 mg and 100 mg / kg body weight. Ten animals were used for each dosage level. All doses were expressed as ibuprofen molar equivalents. The doses used and the molar equivalents are shown below.

The efficacy of the two dosage levels—50.0 and 100.0 mg / kg, and the reference control for the three formulations, as seen from the antagonism of acetylcholine-induced single body torsion, is shown below.

Statistical analysis using the chi-square test did not show any statistically significant differences for the formulation in comparison to the reference control, but for each group, “P” was found to be greater than the significance level of 0.05.

From clinical observations based on the number of animals that did not show torsion due to acetylcholine administration, (±) -ibuprofen-L-hydroxyproline ester was found in other formulations and ibuprofen (racemic) and ibuprofen (S)- Compared to (+), it was found to be more effective at antagonizing acetylcholine-induced twist.

The data was subjected to statistical analysis using the chi-square test to evaluate the efficacy of the new formulation compared to the reference control. The test showed no statistically significant difference for the formulation compared to the reference control, whereas in the comparison of the number of animals that did not show twist in each group, each “P” was a significant level of 0.05. It was found to be larger.

The data is also summarized as FIG. 1 and FIG. From the clinical observation and efficacy comparison bar graphs based on the number of animals showing no twist due to acetylcholine administration (FIGS. 1 and 2), (±) -ibuprofen-L-hydroxyproline ester was found in other formulations and ibuprofen (racemic) and It was found to be more effective at antagonizing acetylcholine-induced torsion than ibuprofen (S)-(+).

Conclusion This study was conducted to evaluate the overall efficacy of a new formulation of ibuprofen. For this purpose, the antagonistic properties of the new preparations on acetylcholine torsion were used as an index for determining the relative efficacy of the preparations. Ibuprofen (racemic mixture and ibuprofen (S)-(+)) served as a reference control. The study was conducted at two dose levels (50.0 mg and 100.0 mg / kg) with the vehicle control group.

The efficacy of the three formulations in terms of antagonistic effects on the two dosage levels—50.0 and 100.0 mg / kg, and the acetylcholine-induced single twist of the reference control is shown below.

Statistical analysis using the chi-square test did not show a statistically significant difference for the formulation in comparison to the reference control, but in the comparison of the number of animals that did not show torsion in each group, each “p "Is found to be greater than the significance level of 0.05.

However, from clinical observations based on the number of animals that did not show torsion due to acetylcholine administration, (±) -ibuprofen-L-hydroxyproline ester was found in other formulations and ibuprofen (racemic) and ibuprofen (S)- Compared to (+), it was found to be more effective at antagonizing acetylcholine-induced twist.

Gastric mucosal irritation potential of L-serine, L-threonine and L-hydroxyproline esters of (±) -ibuprofen in fasted male albino rats

Overview This study shows the relative ability of a new formulation of ibuprofen ((±) -ibuprofen L-serine, L-threonine and L-hydroxyproline ester) to cause gastric mucosal irritation / lesion in fasted male albino rats. Carried out to determine. Ibuprofen (racemic mixture) and ibuprofen (S)-(+) were used as reference controls.

Various new formulations of ibuprofen and ibuprofen (racemic mixture) and ibuprofen (S)-(+) were fasted by gavage using 5% Tween 80 milliQ aqueous solution in the vehicle (F). (Wistar strain). The study was conducted with the vehicle control group at two dosage levels, 200 mg and 300 mg / kg body weight. Five animals were used for each dosage level. All doses were expressed as ibuprofen (racemic mixture) molar equivalents. The doses used and the molar equivalents are shown below.

The various groups used are summarized in the table below.

Rats were fasted for 18-2 hours prior to dosing. The test article was administered as a single dose by gavage. Three hours after drug administration, the animals were sacrificed by CO ₂ gas inhalation. The stomach was excised and observed for the following: amount of mucus exudate, degree of hyperemia and thickening of the stomach wall, bleeding spots (local or pervasive), bleeding characteristics (spotted or spotted bleeding) and size・ Perforation or other lesions

The observation results regarding gastric mucosal irritation of animals in each group are summarized below.

The results of this study showed that none of the ibuprofen formulations provided evidence of irritation to the gastric mucosa of male fasted male albino rats for the two dose levels tested (200 mg and 300 mg / kg body weight). . In contrast, ibuprofen (racemic mixture) and ibuprofen (S)-(+) stimulated the gastric mucosa at the two dose levels tested. Furthermore, ibuprofen (S)-(+) was found to be more irritating to the gastric mucosa than ibuprofen (racemic mixture).

Overview of ketoprofen S (+) threonine ester synthesis:
The procedure for the synthesis of the L-threonine ester of ketoprofen was outlined in the synthetic order section. This synthesis is exemplary and is equally applicable to other amino acids. Complete procedures and analytical data are given in the experimental section. In general, (±) -ketoprofen (5 g) is dissolved in the presence of a catalytic amount of 4- (N, N-dimethylamino) -pyridine (DMAP) in the presence of 1- (3-dimethylaminopropyl) -3- Ethyl carbodiimide hydrochloride (EDC, 1 eq) was used to couple with N-boc-L-threonine t-butyl ester 1 (1 eq). When the reaction was complete, excess EDC was removed by extraction with water, DMAP was removed by extraction with dilute acid, and ketoprofen was removed by extraction with sodium bicarbonate. After drying over sodium sulfate, filtration, and concentration, the crude protected L-threonine- (±) -ketoprofen was purified by flash chromatography on silica gel to give the protected L-threonine ester in high yield (98%). Was generated. The protecting group was removed by treatment with 2M diethyl hydrochloride solution to cleave the boc group followed by treatment with trifluoroacetic acid to remove the t-butyl ester. After drying, the mixture of L-threonine-R, S (±) -ketoprofen ester was separated by crystallization from acetonitrile. The hydrochloride salt of L-threonine-S (±) -ketoprofen ester was preferentially precipitated from acetonitrile. For comparison, an optically pure reference sample was prepared starting from S (±) -ketoprofen. After drying and analysis, a sample of L-threonine-S (±) -ketoprofen ester · hydrochloride (1.75 g) was separated from the mixture.

（±）−ケトプロフェンのＬ−スレオニンの合成：ａ）ＥＤＣ、ＤＭＡＰ、ＣＨ_２Ｃｌ_２；ｂ）ＨＣｌ（２Ｍ）；ｃ）ＴＦＡ；ｄ）ＡＣＮ（結晶化）。 Synthesis order:

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

Experiment chapter:
SP10018A was synthesized in a single batch. Reagents mentioned in the experimental section were purchased from Sigma-Aldrich, Acros or Bachem with the highest purity available except for solvents purchased from Fischer Scientific or Mallinkrodt.

３−［２（Ｒ，Ｓ）−（３−ベンジル−フェニル）−プロピオニルオキシ］−２（Ｓ）−ｔ−ブトキシカルボニルアミノ−酪酸ｔｅｒｔ−ブチルエステル：（ジアステレオマーの混合）

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_３） ： 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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_３） ： 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， １２９．３１， １２９．１３， １２９．０２， １２８．５４， 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． (±) -Ketoprofen (5.32 g, 20.92 mmol), Nt-butylcarbonyl-L-threonine t-butyl ester (Boc-Thr-OtBu, 5.17 g, 18.72 mmol) (prepared 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) ) Was dissolved in dichloromethane (50 mL) at room temperature under an argon atmosphere. After stirring for 5 hours, the dichloromethane layer was washed with water (50 mL), 5% hydrochloric acid (2 × 25 mL), water (25 mL), saturated sodium bicarbonate (2 × 25 mL) and water (50 mL). After drying over sodium sulfate (5 g) for 1 hour, filtering and concentrating under reduced pressure, eluting residual oil (10.3 g) with hexanes / ethyl acetate (2: 1) using silica gel (150 g). Purified by chromatography. After combining the fraction-containing products, it was concentrated to dryness under high vacuum and by this procedure the protected L-threonine (±) -ketoprofen ester SPI001801 (9.42 g, 98% yield) was obtained as a clear oil. Was generated.

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

¹ H NMR (300 MHz, CDC1 ₃ ): 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, CDC1 ₃ ): 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, 129.31, 129.13, 129.02, 128.54, 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.

２（Ｓ）−アミノ−３（Ｒ）−［２（Ｓ）−（３−ベンゾイル−フェニル）−プロピオニルオキシ］−酪酸・塩酸塩（Ｌ−スレオニン−Ｓ（±）−ケトプロフェンエステル・塩酸塩）：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＤＭＳＯ）： δ ＝ １４．０８ （ｂｒ ｓ， １Ｈ）， ８．７２ （ｂｒ ｓ， ３Ｈ）， 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）．

^１３Ｃ 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分析：
ｃａｌｃ．： Ｃ ６１．３０， Ｈ ５．６６， Ｎ ３．５７ ； ｆｏｕｎｄ： Ｃ ６１．０２， Ｈ ５．５８， Ｎ ３．５８．

ＨＰＬＣ分析：
９８．２８％ ｐｕｒｉｔｙ； ｒ． ｔ． ＝ ２５．１４ｍｉｎ．； ５５％ ＤＩＵＦ ｗａｔｅｒ （０．１％ ＴＦＡ）／４５％ ｍｅｔｈａｎｏｌ； １ ｍＬ／ｍｉｎ； ３６．４ Ｃ； Ｌｕｎａ Ｃ１８， ５ｕ ｃｏｌｕｍｎ （ｓｅｒｉａｌ ＃ ２１１７３９−４２）， ４．６ｘ２５０ ｍｍ； ２０ ｕｌ ｉｎｊｅｃｔｉｏｎ．

旋光度： ＋ ２７．０°（２０ Ｃ， １７４．４ ｍｇ／１０ ｍＬ ｅｔｈａｎｏｌ， ５８９ ｎｍ） ； 融点： １６６−１６７ ℃ The protected (R, S) -ketoprofen-L-threonine ester (9.42 g, 18.41 mmol) was dissolved in dichloromethane (25 mL) at room temperature under an argon atmosphere. To this solution was added anhydrous diethyl ether solution (2M, 25 mL) and the mixture continued to stir at room temperature for 17 hours. The mixture was concentrated under reduced pressure. The residual foam (8.2 g) was dissolved in a mixture of dichloromethane (10 mL) and trifluoroacetic acid (20 mL). After stirring at room temperature for 6.5 hours, the solution was concentrated under reduced pressure. Toluene (25 mL) was added to the residual oil and a second mixture concentration was performed. A mixture of ethanol (20 mL) and anhydrous diethyl ether solution (2M, 20 mL) was added, and the solution was concentrated a third time. After drying under high vacuum for 2 hours at room temperature, the experiment was performed as an off-white solid (±) -ketoprofen-L-threonine ester hydrochloride (diastereomer mixture, 7.11 g, crude yield 98%) Was generated. This diastereomeric crude mixture (7.0 g) was crystallized three times from acetonitrile (200 mL). After the third crystallization, the residual solid was dried at 50 ° C. under high vacuum (4 hours) until the weight was constant. The experiment produced L-threonine-S (±) -ketoprofen ester hydrochloride hydrochloride SPI0018A (2.2 g, 30% yield from SPI00181).

2 (S) -amino-3 (R)-[2 (S)-(3-benzoyl-phenyl) -propionyloxy] -butyric acid hydrochloride (L-threonine-S (±) -ketoprofen ester hydrochloride) :

¹ H NMR (300 MHz, DMSO): δ = 14.08 (br s, 1H), 8.72 (br s, 3H), 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; r. t. = 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 ° C.

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_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， ４５．４３， ２８．５３， ２８．１０， １８．９９， １６．９６． Preparation of S- (±) -ketoprofen-L-threonine ester hydrochloride reference material (±) -ketoprofen (1.87 g, 7.74 mmol), Nt-butylcarbonyl-L-threonine t-butyl ester ( Boc-Thr-OtBu, 2.25 g, 8.14 mmol, prepared according to 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) were dissolved in dichloromethane (25 mL) at room temperature under an argon atmosphere. After stirring for 4 hours, the dichloromethane layer was washed with water (25 mL). After drying over sodium sulfate (5 g) for 1 hour, filtration and concentration under reduced pressure, the residual oil was used without purification. This procedure gave protected L-threonine (±) -ketoprofen ester (4.01 g, yield to 100%) as a clear oil.

¹ H NMR (300 MHz, CDC1 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, CDC1 ₃ ): δ = 196.4, 172.79, 168.999, 155.94, 140.44, 137.999, 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, 45.43, 28.53, 28.10 , 18.99, 16.96.

The protected (S) -ketoprofen-L-threonine ester (3.92 g, 7.66 mmol) was dissolved in anhydrous diethyl ether solution (2M, 50 mL) and stirred at room temperature for 17 hours. The mixture was concentrated under reduced pressure. The residual foam (3.4 g) was dissolved in a mixture of dichloromethane (20 mL) and trifluoroacetic acid (20 mL). After stirring at room temperature for 6.5 hours, the solution was concentrated under reduced pressure. Toluene (25 mL) was added to the residual oil and a second mixture concentration was performed. A mixture of ethanol (20 mL) and anhydrous diethyl ether solution (2M, 20 mL) was added, and the solution was concentrated a third time. After drying at room temperature under high vacuum for 2 hours, the experiment produced S (+)-ketoprofen-L-threonine ester hydrochloride (3.05 g, crude material) as an off-white solid. This crude material was stirred with acetone (50 mL) for 2 hours at room temperature under an argon atmosphere. The white residual solid was filtered and dried at 50 ° C. under high vacuum (4 hours) until the weight was constant. The experiment produced L-threonine-S (+)-ketoprofen ester hydrochloride (2.04 g, 67% yield).

¹ H NMR (300 MHz, DMSO): δ = 14.08 (br s, 1H), 8.72 (br s, 3H), 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.

HPLC analysis:
99.43% purity; r. t. = 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.

C. Amino acid derivatives of aspirin
Overview:
Synthetic procedures for L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid are outlined in the synthetic order section and are also exemplary for other amino acids. Complete procedures and analytical data are given in the experimental section. In general, acetylsalicyloyl chloride (10-25 g) was coupled with an N-benzyloxy / benzyl ester protected amino acid in the presence of pyridine. After completion of the reaction (24-48 hours at room temperature), the mixture is poured into ice-cold 2N hydrochloric acid. The dichloromethane fraction was then washed with sodium bicarbonate, water and brine. After drying over sodium sulfate, filtration and concentration, the crude protected amino acid ester of acetylsalicylic acid was purified by flash chromatography on silica gel. This procedure produced protected amino acid esters of acetylsalicylic acid with yields ranging from 68% to 95%. The protecting group was removed by hydrogenation (20 psi H ₂ ) in the presence of 10% palladium on carbon. The amino acid ester of acetylsalicylic acid was extracted from the palladium catalyst with water, concentrated and dried. The final compound was washed with solvent (water, dioxane, acetonitrile, and / or dichloromethane) until pure and dried under high vacuum until the weight was constant.

アセチルサリチル酸のＬ−セリン、Ｌ−スレオニン、およびＬ−ヒドロキシプロリンエステルの合成：ａ）ピリジン、ＣＨ_２Ｃｌ_２；ｂ）１０％Ｐｄ／Ｃ、ＥｔＯＨ、ＥｔＯＡｃ。 Synthesis order:

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.

Experiment chapter:
The synthesis of SPIB00101, SPIB00102 and SPIB00103 was performed in one or two batches. The reagents described in the experimental section were purchased from Lacmaster, Sigma-Aldrich, Acros or Bachem, with the highest purity available, except for solvents purchased from Fischer Scientific or Mallinkrodt.

^１Ｈ 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 （１１Ｈ， 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）．

^１３Ｃ 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， ６７．７２， ６７．２６， ５７．６４， ２０．９８， １６．８８． 1) SPIB00102: 2-O-acetylsalicylic acid (2S, 3R)-(−)-threonine ester N-carbobenzyloxy-L-threonine benzyl ester (Z-Thr-OBzl, 21.77 g) in anhydrous dichloromethane (500 mL) 63.40 mmol) and pyridine (25 mL) were cooled in an ice bath under a nitrogen atmosphere. Acetylsalicyloyl chloride (17.63 g, 88.76 mmol) was added and the mixture was 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 fraction was washed with water (500 mL), saturated sodium bicarbonate solution (500 mL), water (500 mL), brine (500 mL) and then dried over sodium sulfate (25 g). After filtration, it was concentrated under reduced pressure, dried under high vacuum, and the yellow residual oil (35.43 g) was purified using hexanes / acetic acid using silica gel (300 g, 0.035 to 0.070 mm, pore diameter 6 nm). Purified using flash chromatography eluting with ethyl (3: 1). The fraction-containing product was concentrated under reduced pressure and dried under high vacuum until the weight was constant, and the experiment showed that the protected acetylsalicyl-L-threonine ester SPIB0010201 (28.1 g, yield 88) was obtained 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 (11H, 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, CDC 1 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, 67.72, 67.26, 57.64, 20.98, 16.88 .

^１Ｈ ＮＭＲ （３００ ＭＨｚ， Ｄ_２Ｏ−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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， Ｄ_２Ｏ−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分析：
９８．７％ ｐｕｒｉｔｙ； ｒｔ＝ ６．２３３ ｍｉｎ； Ｌｕｎａ Ｃ１８ ５ｕ ｃｏｌｕｍｎ （ｓｎ １６７９１７−１３）； ４．６ｘ２５０ ｍｍ； ２５４ ｎｍ； ３５％ ＭｅＯＨ／６５％ ＴＦＡ （０．１％） ｐＨ＝ １．９５； ３５ Ｃ； ２０ ｕｌ ｉｎｊ．； １ ｍｌ／ｍｉｎ ； ｓａｍｐｌｅ ｄｉｓｓｏｌｖｅｄ ｉｎ ｍｏｂｉｌｅ ｐｈａｓｅ ｗｉｔｈ １ ｄｒｏｐ ｐｈｏｓｐｈｏｒｉｃ ａｃｉｄ．

ＣＨＮ分析：
ｃａｌｃ．： Ｃ ５５．５１， Ｈ ５．３８， Ｎ ４．９８ ； ｆｏｕｎｄ： Ｃ ５５．３７， Ｈ ５．４０， Ｎ ５．０３．

融点： １５３．５ ℃ （ｄｅｃ．） The protected acetylsalicyl-L-threonine ester SPIB0010201 (14.50 g, 28.68 mmol) was dissolved in ethanol (100 mL) and ethyl acetate (100 mL) at room temperature and 10% palladium on carbon (3.0 g, wet rate) To Parr bottle containing 50%) under a nitrogen atmosphere. This nitrogen atmosphere was replaced with hydrogen gas (20 psi). After shaking for 20 hours, the palladium catalyst was removed by filtration through Celite (30 g). The residual solid (palladium / celite and product) was washed with water (600 × 4 mL) until the product was removed. The ethanol fraction and water fraction were concentrated at room temperature under reduced pressure. The residual solid was washed with water (20 mL) and dioxane (20 mL) for 48 hours. After filtration, the white residual solid was dried at room temperature under high vacuum (4 hours, 88 ° C.) until the weight of the product was constant (16 hours). From this experiment, acetylsalicyl-L-threonine ester SPIB00102 (4.40 g, 55% yield) was obtained 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, D ₂ O-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 × 250 mm; 254 nm; 35% MeOH / 65% TFA (0.1%) pH = 1.95 35 C; 20 ul inj. 1 ml / min; sample dissolved in mobile phase with 1 drop 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.)

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_３） ： 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， ｄｄ， Ｊ＝ １１．４， ３．３ Ｈｚ）， ２．３０ （３Ｈ， ｓ）．

^１３Ｃ 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， ６７．８３， ６７．２７， ６４．６３， ５３．５５， ２１．０３． 2) SPIB00101: 2-O-acetylsalicylic acid (2S)-(+)-serine ester N-carbobenzyloxy-L-serine benzyl ester (Z-Ser-OBzl, 23.17 g, 70.34 mmol) and pyridine ( 30 mL) of anhydrous dichloromethane (500 mL) was cooled in an ice bath under a nitrogen atmosphere. Acetylsalicyloyl chloride (21.07 g, 106.1 mmol) was added and the mixture was warmed to room temperature and stirred for 2 days. After 48 hours, the mixture was poured into ice-cold 2N hydrochloric acid (400 mL). After mixing, the layers were separated and the dichloromethane fraction was washed with water (500 mL), saturated sodium bicarbonate solution (500 mL), water (500 mL), brine (500 mL) and then dried over sodium sulfate (25 g). After filtration, it was concentrated under reduced pressure, dried under high vacuum, and a brown residual solid (47.19 g) was obtained using hexanes / ethyl acetate using silica gel (200 g, 0.035 to 0.070 mm, pore size 6 nm). Purified using flash chromatography eluting with (3: 1). The fraction-containing product was concentrated under reduced pressure and dried under high vacuum until the weight was constant, and the experiment showed that the protected acetylsalicyl-L-serine ester SPIB0010101 (32.97 g, yield 95) was obtained as a white solid. %).

¹ H NMR (300 MHz, CDC1 ₃ ): 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, dd, 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, 67.83, 67.27, 64.63, 53.55 , 21.03.

^１Ｈ ＮＭＲ （３００ ＭＨｚ， Ｄ_２Ｏ−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， ｄｄ， Ｊ＝ ４．２， ３．０ Ｈｚ）， ２．３９ （３Ｈ， ｓ）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， Ｄ_２Ｏ−ＤＣ１）： 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分析：
９８．１％ ｐｕｒｉｔｙ； ｒ． ｔ． ＝ ５．８３９ ｍｉｎ．； ６５％ ＴＦＡ （０．１％）／３５％ ｍｅｔｈａｎｏｌ； １ ｍＬ／ｍｉｎ ； ３５ Ｃ； Ｌｕｎａ Ｃ１８， ３ｕ ｃｏｌｕｍｎ （ＳＮ １８４２２５−３７）， ４．６ｘ２５０ ｍｍ； ２２ ｕｌ ｉｎｊｅｃｔｉｏｎ； ＤＡＤ１Ｂ， Ｓｉｇ＝ ２４０， ４ Ｒｅｆ＝ ５５０，１００．

ＣＨＮ分析 ：
ｃａｌｃ．： Ｃ ５３．９３， Ｈ ４．９０， Ｎ ５．２４ ； ｆｏｕｎｄ： Ｃ ５４．０２， Ｈ ５．００， Ｎ ５．２３．

融点： １４７．０ ℃ （ｄｅｃ．） The protected acetylsalicyl-L-serine ester SPIB0010101 (21.0 g, 42.7 mmol) was dissolved in ethanol (100 mL) and ethyl acetate (100 mL) at room temperature and 10% palladium on carbon (4.20 g, wet rate). To a Parr bottle containing 50%) in a nitrogen atmosphere. The nitrogen atmosphere was replaced with hydrogen gas (20 psi). After 5 hours, additional 10% palladium catalyst (4.26 g) was added and the hydrogen atmosphere was returned (2-psi). After further shaking for 20 hours at room temperature, the palladium catalyst was removed by filtration through celite. The residual solid (palladium / celite and product) was washed with water (1500 x 2 mL) until the product was removed. The ethanol fraction and water fraction were concentrated at room temperature under reduced pressure. The residual solid (7.17 g) was dissolved in DIUF water (4.3 L), filtered through celite to remove insoluble material, and then concentrated at room temperature under high vacuum. The white solid was then washed overnight with 1,4-dioxane (100 mL) and DIUF water (50 mL). After 24 hours, the solid was filtered and dried under high vacuum until the weight was constant (24 hours). From this experiment, acetylsalicyl-L-serine ester SPIB00101 (6.17 g, yield 54%) was obtained 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, dd, J = 4.2) 3.0 Hz), 2.39 (3H, s).

¹³ C NMR (75 MHz, D ₂ O-DC1): 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; r. t. = 5.839 min. 65% TFA (0.1%) / 35% methanol; 1 mL / min; 35 C; Luna C18, 3 u column (SN 184225-37), 4.6 × 250 mm; 22 ul injection; DAD1B, Sig = 240, 4 Ref = 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.)

^１Ｈ 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 （１Ｈ， ｍ）， ２．２８ （４Ｈ， ｍ）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_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， １２８．０５， 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． 3) SPIB00103: 2-O- acetylsalicylic acid (2S, 4R) -4- hydroxyproline ester N- carbobenzyloxy -L- hydroxyproline ester (Z-Ser-OBzl, 21.5g , 60.5 ^{mmol) 1} and A mixture of pyridine (25 mL) in anhydrous dichloromethane (500 mL) was cooled in an ice bath under a nitrogen atmosphere. Acetylsalicyloyl chloride (13.2 g, 66.6 mmol) was added and the mixture was warmed to room temperature and stirred overnight. After 24 hours, additional acetylsalicyloyl chloride (5.0 g, 25.2 mmol) was added and stirred overnight. After 48 hours, the mixture was poured into ice-cold 1N hydrochloric acid (500 mL). After mixing, the layers were separated and the dichloromethane fraction was washed with water (500 mL), saturated sodium bicarbonate solution (500 mL), water (500 mL), brine (500 mL) and then dried over sodium sulfate (25 g). After filtration, it was concentrated under reduced pressure, dried under high vacuum, and a yellow residual oil (40.7 g) was added to hexanes / acetic acid using silica gel (460 g, 0.035 to 0.070 mm, pore size 6 nm). Purified using flash chromatography eluting with ethyl (3: 1). The fraction-containing product was concentrated under reduced pressure and dried under high vacuum until the weight was constant, and from the experiment, the protected acetylsalicyl-L-hydroxyproline ester SPIB0010301 (21.31 g, yield) was obtained as a colorless oil. 68%).

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 (1H, m), 2.28 (4H, m).

¹³ C NMR (75 MHz, CDC1 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, 128.05, 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.

^１Ｈ ＮＭＲ （３００ ＭＨｚ， Ｄ_２Ｏ−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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， Ｄ_２Ｏ−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分析：
９８．３％ ｐｕｒｉｔｙ； ｒ． ｔ． ＝ ７．２０１ ｍｉｎ．； ６５％ ＴＦＡ （０．１％）／３５％ ｍｅｔｈａｎｏｌ； １ ｍＬ／ｍｉｎ； ３５ Ｃ ； Ｌｕｎａ Ｃ１８， ３ｕ ｃｏｌｕｍｎ （ＳＮ １８４２２５−３７）， ４．６ｘ２５０ ｍｍ； ２２ ｕｌ ｉｎｊｅｃｔｉｏｎ； ＤＡＤ１Ｂ， Ｓｉｇ＝ ２４０， ４ Ｒｅｆ＝ ５５０，１００．

ＣＨＮ分析：
ｃａｌｃ．： Ｃ ５７．３４， Ｈ ５．１６， Ｎ ４．７８ ； ｆｏｕｎｄ： Ｃ ５７．０９， Ｈ ５．２３， Ｎ ４．９１．

融点： １６２ ℃ （ｄｅｃ．） The protected acetylsalicyl-L-hydroxyproline ester SPIB0010301 (10.6 g, 20.5 mmol) was dissolved in ethanol (75 mL) and ethyl acetate (75 mL) at room temperature and 10% palladium on carbon (3.0 g, wet) To a Parr bottle containing 50%) under a nitrogen atmosphere. The nitrogen atmosphere was replaced with hydrogen gas (20 psi). After shaking for 17 hours at room temperature, the reaction mixture was washed with water (500 mL) for 2 hours. The organic layer (top) was removed using a pipette and the aqueous layer was filtered through celite. The water fraction was concentrated at room temperature under reduced pressure. The residual solid (6.71 g) was then washed overnight with anhydrous dichloromethane (35 mL). After 24 hours, the solid was filtered and dried under high vacuum until the weight was constant (24 hours). From this experiment, acetylsalicyl-L-hydroxyproline ester SPIB00301 (2.87 g, yield 47.7%) was obtained 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; r. t. = 7.201 min. 65% TFA (0.1%) / 35% methanol; 1 mL / min; 35 C; Luna C18, 3 u column (SN 184225-37), 4.6 × 250 mm; 22 ul injection; DAD1B, Sig = 240, 4 Ref = 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 ° C. (dec.)

Gastric mucosal irritation ability of L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid as compared to acetylsalicylic acid:
This study is to determine the relative potential that a novel formulation of aspirin (L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid) causes gastric mucosal irritation / injury in fasted male albino rats Implemented. Aspirin was useful for reference control.

A variety of new aspirin formulations and aspirin were prepared by fasting male albino rats by gavage using 0.5% (w / v) carboxymethylcellulose (CMC) phosphate buffer (pH 2.6) as a vehicle. (Wistar strain). This study was conducted with the vehicle control group at two dose levels: 100 mg and 200 mg / kg body weight. Five animals were used for each dose level. All doses were expressed as aspirin molar equivalents. The doses used as well as the molar equivalents are presented below.

Rats were fasted for 18-22 hours prior to dosing. The test article was administered as a single dose by gavage. Three hours after drug administration, the animals were sacrificed by CO ₂ gas inhalation. The stomach was excised and observed for the following: amount of mucus exudate, degree of hyperemia and thickening of the stomach wall, bleeding spots (local or widespread), bleeding characteristics (spot or spotted bleeding) and size・ Perforation

The observations regarding gastric mucosal irritation in various groups of animals are summarized below.

In conclusion, acetylsalicylic acid L-serine, L-threonine and L-hydroxyproline ester showed no evidence of irritation of the gastric mucosa at the two doses tested, namely 100 and 200 mg / kg body weight. In contrast, aspirin (acetylsalicylic acid) stimulated the gastric mucosa of all fasted male albino rats at a dose level of 200 mg / kg. However, aspirin showed no evidence of irritation to the gastric mucosa of male rats at a dose level of 100 mg / kg.

Furthermore, none of the animals in the various test groups showed clinical signs of toxicity throughout the 3 hour observation period.

Efficacy of L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid on clotting time in rats as measured 1 hour after dosing compared to acetylsalicylic acid

Observation of blood clotting time The mean clotting time (MCT) of the animals in the low, medium and high dose groups, vehicle control group and positive control group of various formulations measured 1 hour after dosing is presented below (Table 14):

Figures 3-6 illustrate animal group mean data for mean clotting time expressed as dose relationship + minutes for L-serine ester of aspirin and control.

Statistical analysis showed a significant improvement in efficacy at the 5% significant level when compared to the vehicle control group, with the high and medium doses (FIG. 7).

FIG. 4 shows group average data for animals. Figure 2 shows the dose-response relationship of mean clotting time (MCT) in minutes for L-hydroxyproline ester of aspirin. The statistical analysis in FIG. 4 showed a significant improvement of 5% for the high and low dose efficacy when compared to the vehicle control group (FIG. 6).

FIG. 5 shows the dose response relationship of mean clotting time (MCT) in minutes for the L-threonine ester of acetylsalicylic acid. Statistical analysis showed a significant improvement of 5% significant level for high dose efficacy when compared to vehicle control.

FIG. 6 shows the dose-response relationship of the average clotting time of acetylsalicylic acid. Statistical analysis showed a significant improvement of 5% significant level for medium and high dose efficacy when compared to vehicle control (Figure 7). The dose-response efficacy was statistically significant and apparent without question.

CONCLUSION This study was conducted to evaluate the efficacy of a new formulation of aspirin using blood clotting time as an index in albino rats. Aspirin served as a positive control. This study was conducted for 3 dose levels using the new formulation and positive control with the vehicle control group.

Dose The main study dose was selected based on a dose range search experiment with acetylsalicylic acid. All doses were expressed as aspirin molar equivalents. The doses used in the main experiments for the various formulations and positive controls were the same and are presented below.

Efficacy (blood clotting time)
The efficacy as seen from the time required for blood clotting time at various dosage levels and various dosage levels of acetylsalicylic acid-low, medium and high doses is presented below.

L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid are as significant as acetylsalicylic acid in terms of coagulation time observed after 1 hour, but are less irritating to the stomach than acetylsalicylic acid As for any level, it was much better.

Efficacy of L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid on clotting time in rats as measured 2 hours after dosing compared to acetylsalicylic acid. Evaluation of the efficacy of L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid when blood coagulation time measured at ± 10 minutes was used as an indicator in albino rats compared to acetylsalicylic acid Went for. Aspirin served as a positive control. Male albino rats were exposed to aspirin and three new aspirin formulations at a single dose level of 20 mg / kg body weight. The vehicle control group was not used. The dose was expressed as aspirin molar equivalents. The doses used in the main experiments for the various formulations and positive controls are as follows.

Efficacy (blood clotting time)
The efficacy of various formulations and aspirin (positive control) as seen from the time required for blood clotting time at a dose level of 20 mg / kg body weight is presented below.

Observation of blood clotting time The data regarding the mean clotting time (MCT) of the animals, measured at 2 hours after dosing for various formulations, vehicle control and positive control at a dose level of 20 mg / kg body weight are presented below.

L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid have been found to be effective in clotting time.

In conclusion, based on the time required for blood to clot (coagulation time) measured 2 hours after dosing, the amino acid prodrug was observed to have efficacy. However, the L-threonine ester of acetylsalicylic acid was found to have a higher relative potency compared to the other two formulations.

As shown in FIG. 7, statistical analysis shows that L-threonine and L-hydroxyproline esters of acetylsalicylic acid are as effective as acetylsalicylic acid, and that L-hydroxyproline ester of acetylsalicylic acid and L-threonine ester of acetylsalicylic acid are The mean blood clotting time observed after 2 hours shows no significant difference of 5% from the positive control. However, when combined with gastric irritation, the L-serine, L-threonine and L-hydroxyproline esters of acetylsalicylic acid are far superior.

There are a variety of screening tests that determine the usefulness of prodrugs made according to the disclosed methods. They include both in vitro and in vivo screening methods.

In vitro methods include acid / base hydrolysis of prodrugs, hydrolysis in porcine pancreas, hydrolysis in rat intestinal fluid, hydrolysis in human gastric fluid, hydrolysis in human intestinal fluid and hydrolysis in human plasma. Can be mentioned. These assays include: Simmons, DM, Chandran, VR and Portmann, GA, Danazol Amino Acid Prodrugs, 68: In Vitro and In Situ Biopharmaceutical Evaluation, Drug Development. The entire contents of which are incorporated by reference.

The compounds of the present invention are effective in the treatment of diseases or conditions for which NSAIDs are usually used. The prodrugs disclosed herein enhance the therapeutic benefits of NSAIDs by deforming in the body to release the active compound and reduce or eliminate the biopharmaceutical and pharmacokinetic disorders associated with each NSAID. However, it will be necessary to note that these prodrugs have sufficient activity per se in mammals without releasing the active agent. Prodrugs are more water soluble than ibuprofen or other NSAIDs, so they do not need to be associated with a carrier vehicle that can cause harmful or unwanted side effects such as alcohol or castor oil. Furthermore, oral formulations containing NSAID prodrugs are absorbed by blood and are extremely effective.

Thus, the prodrugs of the present invention enhance the therapeutic benefit by removing the biopharmaceutical and pharmacokinetic obstacles of existing drugs.

Furthermore, prodrugs are easily synthesized in high yield using commercially available reagents.

IV. Summary of proline derivatives of acetaminophen:
The procedure for the synthesis of L-proline ester of acetaminophen is outlined in the synthetic order section. The synthesis is exemplary. Complete procedures and analytical data are given in the experimental section. Acetaminophen (10 g) was coupled with Boc-L-proline using EDC in the presence of a catalytic amount of DMAP. After the reaction was complete (3 hours at room temperature), the solution was washed with water. After drying, filtering and concentrating through sodium ester sulfate, the crude amino acid ester of acetaminophen was purified by flash chromatography on silica gel. This procedure produced a protected L-proline ester of acetaminophen at a rate of 72%. The protecting group was removed by dissolving the ester in dichloromethane and passing hydrogen chloride through the solution at room temperature. After filtration, the final salt was stirred in tetrahydrofuran until pure. The yield of the deprotection step was 91.4% after filtration and drying under high vacuum at 90 ° C. for 4 hours.

アセトアミノフェンのＬ−プロリンエステルの合成：ａ）ＥＤＣ、ＤＭＡＰ、ＣＨ_２Ｃｌ_２；ｂ）ＨＣｌ（ｇ）、ＣＨ_２Ｃｌ_２。 Synthesis order:

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

Experiment chapter:
The synthesis of SPI0014 was performed in one batch. The reagents described in the experimental section were purchased from Lancaster, Sigma-Aldrich or Acros with the highest purity available, except for solvents purchased from Fisher Scientific or Mallinkrodt.

^１Ｈ 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 （２ Ｈ， ｍ）， ４．４７ （１ Ｈ， ｍ）， ３．５９−３．４５ （２Ｈ， ｍ）， ２．３６ （１ Ｈ， ｍ）， ２．１７−１．９０ （６ Ｈ， ｍ）， １．４６ （９ Ｈ， ｍ）．

^１３Ｃ 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， ２４．２８， ２３．７８． SPI0014: pyrrolidine-2 (S) -carboxylic acid 4-acetylamino-phenyl ester / hydrochloride Boc-L-proline (14.39 g, 68.80 mmol), acetaminophen (10.02 g, 66.28 mmol) , EDC (12.9 g, 67.29 mmol) and DMAP (1.10 g, 9.0 mmol) in anhydrous dichloromethane (100 mL) were stirred at room temperature under an argon atmosphere for 3 hours. After 3 hours, water (120 mL) was added. After mixing for 5 minutes, the layers were separated and the dichloromethane fraction was washed with water (120 mL) and dried over sodium sulfate (5 g). After filtration, concentration under reduced pressure, and drying under high vacuum, the residual oil (24.10 g) was used on silica gel (100 g, 0.035 to 0.070 mm, pore diameter 6 nm) using hexanes / ethyl acetate (1: 2). And purified by flash chromatography eluting with The fraction-containing product was concentrated under reduced pressure and dried under high vacuum until the weight was constant. From the experiment, the protected acetaminophen-L-proline ester SPI001401 (16.71 g) was obtained as a white solid (foam). Yield 72.3%).

¹ H NMR (300 MHz, CDC 1 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 (2 H, m), 4.47 (1 H, m), 3.59-3 .45 (2H, m), 2.36 (1 H, 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, 24.28, 23.78.

The protected acetaminophen-L-proline ester SPI001401 (16.60 g, 47.64 mmol) was dissolved in dichloromethane (400 mL) and hydrogen chloride gas was passed through the solution for 2 hours at room temperature. The residual solid was sunk (1 hour). From the white precipitate, dichloromethane was carefully decanted. Tetrahydrofuran (200 mL) was added to the precipitate and the mixture was stirred for 2 hours under an argon atmosphere. After filtration, the white residual solid was dried under high vacuum at 90 ° C. until the product weight was constant (4 hours). From this experiment, acetaminophen-L-proline ester / hydrochloride SPI00140 (12.4 g, yield 91.4%) was obtained as a white solid.

¹ H NMR (300 MHz, CDCL ₃ -DMSO): δ = 10.41 (1H, brs), 10.26 (1H, s), 9.55 (1H, brs), 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, 3H), 2.04 (2H, 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 × 250 mm; 254 nm; 15% MeOH / 85% hexane sulfate buffer (110 mMol, pH = 6 mMol, pH = 35 mM) C; 20 ul inj. 1 ml / min; 5 mg / mL sample 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 ° C

式中ＭｅＢｍｔは、次式のＮ−メチル−（４Ｒ）−４−ブト−２Ｅ−エン−１−イル−４−メチル−（Ｌ）スレオニル残基である

（式中、−ｘ−ｙ−はＣＨ＝ＣＨ−（トランス）である）。その他類似の生成物としては、シロリムス（ｂ）、タクロリムス（ｃ）およびピメクロリムス（ｄ）が挙げられ、それぞれ次の構造を有する：

V. Amino acid derivatives of cyclosporin A Macrocyclic immunosuppressive agents are structurally characteristic, a class of cyclic poly N-methylated undecaptides, and generally have pharmacological, especially immunosuppressive, anti-inflammatory and / or antiparasitic activity Having similar semi-synthetic macrolide structures. The first isolated cyclosporin is the naturally occurring fungal metabolite cyclosporin (Ciclosporin or Cyclosporine), also known as cyclosporin A, which has the following chemical formula:

Where MeBmt is an N-methyl- (4R) -4-but-2E-en-1-yl-4-methyl- (L) threonyl residue of the formula

(Wherein -x-y- is CH = CH- (trans)). Other similar products include sirolimus (b), tacrolimus (c) and pimecrolimus (d), each having the following structure:

Therefore, this class of cyclosporines today is very large, eg [Thr] ² −, [Val] ² −, [Nva] ² −, and [Nva] ² [Nva] ⁵ -cyclosporin (respectively cyclosporin C , D, G and M), [Dihydrop-MeBmt] ^1- [Val] ² -Cyclosporin (also known as Dihydro-Cyclosporin D), [(D) Ser] ⁸ -Cyclosporine, [MeIle] ¹¹ -Cyclosporine, [(D) MeVal] ¹¹ -cyclosporin (also known as cyclosporin H), [MeAla] ⁶ -cyclosporin, [(D) Pro] ³ -cyclosporine, and the like.

In accordance with conventional cyclosporine nomenclature, they are defined with reference to the structure of cyclosporine (ie, cyclosporin A) throughout the specification and claims. This is the first indication of an amino acid residue that is different from the amino acid residue present in the cyclosporine (eg, “[(D) Pro] ³ ” indicates that the cyclosporin in question is in the 3-position-Sar-residue. Without-(D) Pro-), then the term cyclosporin is used to characterize the remaining residues that are identical to those present in cyclosporin A.

As used herein, the term “cyclosporine” refers to various types of cyclosporine, where xy in the MeBmt residue has cis or trans CH═CH, or in which Are also included in derivatives in which one or more amino acids at positions 2-11 of cyclosporin A are replaced with other amino acids. However, in the formula of cyclosporin A, the number of amino acids replaced is preferably 2 or less, and more preferably 1 or less amino acids are replaced.

In addition, the amino acid residues represented by abbreviations, such as -Ala-, -MeVal- and aAbu, are not specifically indicated according to conventional practice, for example as in "-(D) Ala-". As long as it has (L) -type. In the case of abbreviations preceded by “Me”, as in “-MeLeu-”, they represent α-N-methylated residues. Individual residues of the cyclosporine molecule are numbered clockwise, starting at position 1, such as residues -MeBmt-, dihydro-MeBmt-, etc., according to convention in the art. Throughout the specification and claims, the same numerical sequence is used.

Due to its unique medicinal properties, macrocyclic immunosuppressants have attracted much attention in the media. The term “macrocyclic immunosuppressant” includes various natural and semi-synthetic derivatives of cyclosporine and other macrolides such as sirolimus, tacrolimus and pimecrolimus. The main areas of clinical research of the above drugs are as immunosuppressants, especially organ transplants such as heart, lung, heart-lung combinations, liver, kidney, pancreas, bone marrow, skin and corneal transplants, especially heterogeneous genotype organs Related to recipient application in transplantation. These agents are also used to treat psoriasis, atopic dermatitis, rheumatoid arthritis and nephritic syndrome.

Macrocyclic immunosuppressants further include inflammation with a variety of autoimmune diseases and inflammatory conditions, in particular pathogenesis including autoimmune components such as arthritis (eg rheumatoid arthritis, progressive chronic arthritis and osteoarthritis) and rheumatic diseases. Very useful in treating the condition. Specific autoimmune diseases for which cyclosporine treatment has been proposed or applied include autoimmune blood disorders (eg hemolytic anemia, aplastic anemia, true erythrocytic anemia and idiopathic thrombocytopenia), Systemic lupus erythematosus, polychondritis, scleroderma, Wagner granulomas, dermatomyositis, chronic active hepatitis, myasthenia gravis, psoriasis, Steven Johnson syndrome, idiopathic sprue, including ulcerative colitis and Crohn's disease Autoimmune inflammatory bowel disease, endocrine eye disease, Graves' disease, sarcoidosis, multiple sclerosis, primary biliary cirrhosis, juvenile diabetes (type I diabetes), uveitis (anterior and posterior), dry horn Conjunctivitis and spring keratoconjunctivitis, interstitial pulmonary fibrosis, psoriatic arthritis, atopic dermatitis, and nephritis (eg, sudden nephrotic syndrome) Others include those with nephrotic syndrome, and without one) containing minimal change disease.

Furthermore, macrocyclic immunosuppressive agents are also applicable as antiparasitic agents, particularly antiprotozoal agents, and have been suggested to be useful in the treatment of malaria, coccidioidomycosis and schistosomiasis. Recently, they have been taught to be useful as agents such as for the regression or invalidation of antineoplastic resistant tumors.

Although macrocyclic immunosuppressants make a significant contribution, there are difficulties in providing a more effective and convenient means of administration (eg, gallen formulations, eg, convenient for the patient while at the same time providing adequate bioavailability Oral dosage form that can be administered at an appropriate and controlled dosage rate), as well as the reported undesirable side effects: especially the occurrence of nephrotoxic reactions has become a major obstacle to wider use or application .

Furthermore, the macrocyclic immunosuppressive agent described above exhibits strong hydrophobicity as a characteristic and easily precipitates even when a very small amount of water is present, for example, just by touching the body (for example, gastric juice). Therefore, it is extremely difficult to provide an oral formulation that can be tolerated by patients, for example in terms of shape and taste, can be stored, can be administered on a regular basis, and can be administered appropriately and managed by the patient.

Therefore, the proposed liquid formulations, for example for oral administration of macrocyclic immunosuppressants, are here mainly based on similar excipients such as ethanol and oil or carrier medium. Therefore, commercially available macrocyclic immunosuppressive beverages use ethanol and olive oil or corn oil as a carrier medium, eg, in combination with a solvent system containing ethanol and LABRIFIL and equivalent excipients as a carrier medium. . Accordingly, commercially available macrocyclic immunosuppressive beverages use ethanol and olive oil or corn oil in combination as a carrier medium and Labrifil as a surfactant. See, for example, US Pat. No. 4,388,307. However, the use of beverages and similar compositions proposed in the art is associated with various difficulties.

Furthermore, it has been found that there is a problem with the palatability of known oil-based systems. The taste of known beverages is particularly unpleasant. It is widely practiced to dilute large amounts of a suitable flavored beverage, such as a chocolate beverage, just prior to ingestion in order to provide all the usual therapy that is acceptable. Employing an oil-based system also requires the use of high concentrations of ethanol, and alcohol itself is not desirable, especially if it is anticipated for administration to children. In addition to this, evaporation of ethanol from, for example, capsules (as widely discussed to address palatability issues, as discussed) or other shapes (eg, open-type) can lead to macrocyclic immunosuppressive agents. This causes precipitation to proceed. An additional problem arises when such compositions are encapsulated in, for example, soft gelatin. Packaging encapsulated products in hermetic parts, such as hermetic blisters or aluminum foil blister packaging, is particularly difficult. In other words, this makes the product bulky and increases manufacturing costs. Moreover, the storage characteristics of the formulation are far from ideal.

The level of bioavailability obtained when using existing oral macrocyclic immunosuppressive drug delivery systems is also low, showing significant variation at different times during the therapy period in individuals, individual patient types, and even single individuals. The literature reports that the average absolute bioavailability is only about 30% with currently available treatments using currently commercially available macrocyclic immunosuppressive beverages, and between individual groups, for example the liver It shows significant variation between transplant recipient groups (relatively low bioavailability) and bone marrow (relatively high bioavailability). The reported variation in bioavailability among subjects is between 1 or several percent in some patients but reaches up to 90% or more in other patients. As already mentioned, it is often observed that individual availability varies greatly with time. Accordingly, there is a need for uniform and high bioavailability of macrocyclic immunosuppressants in patients.

The use of such dosage forms is also characterized by extremely variable patient doses required. In order to achieve effective immunosuppressive therapy, the blood or serum levels combined from cyclosporine must remain constant for a period of time. This need also depends on the specific condition being treated, for example whether the treatment is for prevention of transplant rejection or for the management of an autoimmune disease or condition, and the immunity of formulas a to d. It varies greatly depending on whether alternative immunosuppressive therapies are accompanied by any of the inhibitors. With normal dosage forms, the resulting bioavailability levels vary widely, so the daily dosage required to obtain the required serum levels will also vary widely between individuals and even within the same individual. . This requires regular and frequent monitoring of blood / serum levels in patients undergoing macrocyclic immunosuppressant therapy. Blood / serum level monitoring is generally performed by RIA or equivalent immunoassay techniques, for example using techniques based on monoclonal antibodies, and must be performed periodically. Inevitably, monitoring is time consuming, inconvenient and greatly increases the overall cost of treatment.

The blood / serum levels of macrocyclic immunosuppressants obtained when using available dosing systems show very large variations in levels between peaks and valleys. That is, in each patient, the level of effective macrocyclic immunosuppressant in the blood varies greatly between administrations of individual medications.

Macrocyclic immunosuppressants are also required to be in water-soluble form for injection. Cremephore L, which is currently used in macrocyclic immunosuppressants, is a polyoxyethylated derivative of castor oil and is well known to be a toxic vehicle. There are many anaphylactic accidents caused by castor oil components. Due to the low water solubility of macrocyclic immunosuppressants, there are currently no formulations that allow macrocyclic immunosuppressants to be present in aqueous solutions at the required concentration.

Above all, a very clear practical difficulty is that the occurrence of undesirable side effects is observed when using the previously mentioned oral dosage forms available.

There have been several proposals in the art to address these various problems, including solid and liquid oral dosage forms. However, macrocyclic immunosuppressive agents are inherently insoluble in aqueous media and can therefore be used conveniently, while at the same time meeting the required criteria for bioavailability, allowing for effective reabsorption from the stomach or gastrointestinal lumen, for example. The most important problems remain unresolved: the use of dosage forms that can contain macrocyclic immunosuppressants in sufficiently high concentrations, which can achieve adequate high blood / serum levels without fluctuations, ing.

A particular difficulty experienced in connection with oral administration of macrocyclic immunosuppressive agents is that the use of macrocyclic immunosuppressive agents is inevitably limited for the treatment of disease states of relatively low severity or risk. is there. Specific areas of difficulty in this aspect are treatment of autoimmune diseases and other conditions that affect the skin, such as treatment of atopic dermatitis and psoriasis, and hair growth as widely proposed in the art. For example, macrocyclic immunosuppressant treatment is used to promote alopecia due to aging or disease.

For example, although oral macrocyclic immunosuppressant treatment has been shown to have great potential benefits for the drug, for example, in patients suffering from psoriasis, the risk of side effects following oral treatment precludes general use. Various proposals have been made in the art for the application of macrocyclic immunosuppressive agents, for example, local forms and various local delivery systems are described. However, topical application attempts have failed to provide a clear and effective treatment.

However, the present invention overcomes the above problems. More specifically, embodiments of the present invention significantly enhance solubility in aqueous solutions, thereby eliminating the need for using carriers such as ethanol or castor oil when administered as a solution. Is a prodrug of the drug. Furthermore, the prodrug of the macrocyclic immunosuppressant of the present invention does not show the side effects of conventional preparations. Still further, the present inventors have shown that the macrocyclic immunosuppressant prodrug of the present invention enhances its absorption when administered to a patient in the form of a prodrug, thereby greatly enhancing its bioavailability and its efficacy. I understood that.

Accordingly, in one embodiment, the present invention is directed to prodrugs of macrocyclic immunosuppressive agents. Prodrugs consist of amino acids esterified to any one hydroxyl group of cyclosporine, sirolimus, free hydroxy groups on the side chain of tacrolimus, and pimecrolimus.

またはその薬学的に許容される塩を目的とし、
式中ＣＹＣＬＯはシクロスポリン分子の位置２乃至１１にある残基を表し、ｘ−ｙはＣＨ＝ＣＨまたはＣＨ_２ＣＨ_２であり、ＡＡはアミノ酸もしくは式ＧＬＹ−ＡＡのジペプチドを表す。後者の場合、ＧＬＹはグリシンを、ＡＡはいずれかのα−アミノ酸である。ジペプチド構造では、ＡＡはグリシンをスペーサとして用い、ＯＨ基を介して薬剤に付加している。グリシンをシクロスポリンにエステル化してから、グリシンのアミノ基とＡＡのカルボキシル酸基を用いたアミド連結によりグリシンをＡＡに結合する。 For example, one aspect of the present invention is a compound of the formula

Or for a pharmaceutically acceptable salt thereof,
CYCLO wherein represents a residue located 2 to 11 of the cyclosporine molecule, x-y is CH = CH or _CH 2 CH _2, AA represents a dipeptide amino acid or formula GLY-AA. In the latter case, GLY is glycine and AA is any α-amino acid. In the dipeptide structure, AA uses glycine as a spacer and is attached to the drug via the OH group. After glycine is esterified to cyclosporine, glycine is bound to AA by amide linkage using the amino group of glycine and the carboxylic acid group of AA.

The present invention is further directed to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formulas ad above and a pharmaceutical carrier therefor.

In another embodiment, the present invention is directed to a method of treating a patient in need of macrocyclic immunosuppressant treatment, the method comprising administering to said patient an effective amount of a compound of formula ad. Including.

In a further embodiment, the present invention provides the hydroxy functionality in the MeBmt moiety at position 1 of the cyclosporine molecule as well as the specific hydroxyl group of formulas b-d and the amino acid or acylated derivative thereof under ester forming conditions. Macrocyclic immunity to aqueous solutions comprising reacting or reacting with a dipeptide structure using a simple amino acid or glycine as a spacer and adding AA to the drug, and isolating the product It aims at the method of improving the solubility of an inhibitor.

In an even further embodiment, the present invention provides for reacting the hydroxy functionality of the MeBmt moiety at the cyclosporin molecule position with an amino acid or acylated derivative thereof under ester-forming conditions, as well as certain hydroxyls of formula bd Reaction of amino acids with amino acids or acylated derivatives thereof under ester-forming conditions, or reaction with a dipeptide structure in which AA is added to the drug using a simple amino acid or glycine as a spacer, formation thereof It is aimed at a method for increasing the bioavailability of a macrocyclic immunosuppressant administered to a patient comprising isolating the product and administering the product to the patient.

Overview:
The procedure for the synthesis of N- (L-proline) -glycine and N- (L-lysine) -glycine ester of cyclosporin A is outlined in the synthetic order section. These examples are illustrative of synthetic schemes using amino acids. Complete procedures and analytical data are given in the experimental section. Cyclosporine A (15 g) was coupled with chloroacetic anhydride (4 eq) in anhydrous pyridine. From the experiment, cyclosporin A chloroacetate SPI001201 (14 g, yield 88%) was obtained in high yield. The chloroacetic acid ester (10.1 g) was then treated with sodium azide in DMF to produce cyclosporin A azidoacetate ester SPI001202 (9.9 g, 97% yield). Next, azidoacetate (9.8 g) was reduced with tin chloride (9 g) to prepare cyclosporin A glycine ester (8.54 g, yield 89%). The glycine ester SPI001203 of cyclosporin A was then coupled with a 2-fold excess of boc-L-proline or Boc-L-lysine using EDC as the coupling agent. After purification by column chromatography, the boc protecting group was removed from the dipeptide ester of cyclosporin A by treatment with 2M hydrochloric acid in diethyl ether at low temperature (5 ° C.). The L-lysine-glycine ester salt of cyclosporin A did not require further purification and was dried. The L-proline-glycine ester salt of cyclosporin A required purification. The salt was converted to the free base using sodium bicarbonate and then filtered through silica gel (acetone elution) and purified. The salt was then formed using anhydrous hydrochloric acid diluted at low temperature and dried.

シクロスポリンＡのＮ−（Ｌ−プロリン）−グリシンおよびＮ−（Ｌ−リジン）−グリシンエステルの合成：ａ）ピリジン；ｂ）ＮａＮ_３、ＤＭＦ；ｃ）ＳｎＣｌ_２、メタノール；ｄ）ｂｏｃ−Ｌ−リジン、ＥＤＣ；ｅ）ｂｏｃ−Ｌ−プロリン、ＥＤＣ；ｆ）ＨＣｌ、Ｅｔ_２Ｏ。 Synthesis order:

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

Experiment chapter:
The synthesis of SPI0022 and SPI0023 was performed in batches. In general, small scale experiments were performed first, followed by larger batches. The reagents described in the experimental section were purchased from Aldrich, Across or Bachem with the highest purity available except for solvents purchased from Fischer Scientific or Mallinckrodt. Cyclosporin A (USP grade) used in these procedures is available from Signature Pharmaceuticals, Inc. Offered by

シクロスポリンＡ（１５．０１ｇ、０．０１２４ミリモル）を、室温、アルゴン雰囲気下で無水ピリジン（３５ｍｌ）に溶解した。溶液を、氷／水槽内で５℃まで冷却してから、無水クロロ酢酸（９．１０ｇ、０．０５３ミリモル）を加えた。１０分間攪拌した後、氷槽を取り除き、溶液を、窒素雰囲気下、室温で１７時間攪拌し続けた。１７時間後、ジエチルエーテル（２００ｍＬ）を加えた。エーテルを水（２×１００ｍＬ）で洗浄してから、硫酸ナトリウム（１０ｇ）を通して１時間乾燥した。濾過および減圧下で濃縮した後、黄色の残留発泡体を高真空下に乾燥し（室温で１時間）、シリカゲル（２００ｇ）を用い、ヘプタン／アセトン（２：１）で溶出するフラッシュクロマトグラフィーにより精製した。留分含有生成物を組み合わせてから濃縮し、最後に、黄色の残留発泡体（１４．８ｇ）を高温のジエチルエーテル（１４０ｍＬ）から結晶化して精製した。冷却（−１０℃、２時間）、濾過および高真空下で乾燥したところ、この手順から白色の固体として、シクロスポリンＡのクロロ酢酸エステルＳＰＩ００１２０１（１４．０ｇ、収率８８．３％）を得た。
シクロスポリンＡクロロ酢酸エステル：

^１Ｈ 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， １５ Ｈ）， 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）．

^１３Ｃ 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， ５５．８０， 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， ２３．９９， 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． 1) SPI001201

Cyclosporine A (15.01 g, 0.0124 mmol) was dissolved in anhydrous pyridine (35 ml) at room temperature under an argon atmosphere. The solution was cooled to 5 ° C. in an ice / water bath before chloroacetic anhydride (9.10 g, 0.053 mmol) was added. After stirring for 10 minutes, the ice bath was removed and the solution was kept stirring at room temperature for 17 hours under a nitrogen atmosphere. After 17 hours, diethyl ether (200 mL) was added. The ether was washed with water (2 × 100 mL) and then dried over sodium sulfate (10 g) for 1 hour. After filtration and concentration under reduced pressure, the yellow residual foam is dried under high vacuum (1 hour at room temperature) and purified by flash chromatography using silica gel (200 g) eluting with heptane / acetone (2: 1). Purified. The fraction-containing products were combined and concentrated, and finally the yellow residual foam (14.8 g) was purified by crystallization from hot diethyl ether (140 mL). Cooling (−10 ° C., 2 hours), filtration and drying under high vacuum yielded chloroacetic acid ester SPI001201 (14.0 g, 88.3% yield) of cyclosporin A as a white solid from this procedure. .
Cyclosporine A chloroacetate:

¹ H NMR (300 MHz, CDC 1 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, 15 H), 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, 53H).

¹³ C NMR (75 MHz, CDC 1 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, 55.80, 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, 23.99, 23.75, 22.85, 21.94, 21.41, 21.25, 20. 84, 1 .85, 18.79, 18.32, 17.89, 17.82, 15.46, 15.24, 10.08.

シクロスポリンＡのクロロ酢酸エステルＳＰＩ００１２０１（１０．０１ｇ、７．８９ミリモル）を、無水Ｎ，Ｎ−ジメチルホルムアミド（３０ｍＬ）に、室温で溶解した。アジ化ナトリウム（２．１５ｇ、３３．０ミリモル）を加えた。混合物を室温で２４時間、アルゴン雰囲気下で暗所で攪拌し続けた。２４時間後、ジエチルエーテル（１５０ｍＬ）を加え、沈殿物を濾過した。エーテルを水（２×１００ｍＬ）で洗浄し、硫酸ナトリウム（１５ｇ）を通して３０分間乾燥、濾過してから、減圧下で濃縮した。残留白色の固体を高真空下で１時間、室温で乾燥した。この実験から、白色の固体としてシクロスポリンＡのアジドアセテートエステルＳＰＩ００１２０２（９．９０ｇ、収率９７％）が生成され、それをそれ以上精製せずに使用した。
シクロスポリンＡアジドアセテートエステル：

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_３）：
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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_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， ２４．６７， 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． 2) SPI001202

Cyclosporin A chloroacetate ester SPI001201 (10.01 g, 7.89 mmol) was dissolved in anhydrous N, N-dimethylformamide (30 mL) at room temperature. Sodium azide (2.15 g, 33.0 mmol) was added. The mixture was kept stirring in the dark under an argon atmosphere for 24 hours at room temperature. After 24 hours, diethyl ether (150 mL) was added and the precipitate was filtered. The ether was washed with water (2 × 100 mL), dried over sodium sulfate (15 g) for 30 minutes, filtered, and concentrated under reduced pressure. The residual white solid was dried at room temperature under high vacuum for 1 hour. This experiment produced the azidoacetate ester SPI001202 (9.90 g, 97% yield) of cyclosporin A as a white solid, which was used without further purification.
Cyclosporine A azide acetate ester:

¹ H NMR (300 MHz, CDC 1 ₃ ):
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, 53H).

¹³ C NMR (75 MHz, CDC 1 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, 24.67, 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.

シクロスポリンＡのアジドアセテートエステルＳＰＩ００１２０２（９．８０ｇ、７．６２ミリモル）を、メタノール（２５０ｍＬ）に室温で溶解した。水（４０ｍＬ）を加え、続いて塩化第一スズ（５ｇ、２６．３ミリモル）を加えた。追加量（４ｇ、２１．０ミリモル）の塩化第一スズ溶液を加え、１時間、室温で攪拌し続けた。溶液をさらに２時間、室温で攪拌した。水酸化アンモニウム（４０ｍＬ、２９％）を含有する水（２００ｍＬ）を加えた。濾過後、溶液を減圧下に濃縮した（２００ｍＬまで）。残留水溶液を酢酸エチル（２×２００ｍＬ）で抽出した。酢酸エチル留分を組み合わせ、硫酸ナトリウム（２０ｇ）を通して乾燥、濾過してから減圧下で濃縮した。透明な残留発泡体を、シリカゲル（１５０ｇ）で濾過し、ジクロロメタン／メタノール（２：１）で溶出して精製した。この手順から、透明な固体の発泡体として、シクロスポリンＡのグリシンエステル（８．５４ｇ、収率８９％）を得た。
シクロスポリンＡのグリシンエステル：

^１Ｈ 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， ５３ Ｈ）．

^１３Ｃ 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， ５４．７４， 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， ２４．５４， 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． 3) SPI001203

Cyclosporine A azidoacetate ester SPI001202 (9.80 g, 7.62 mmol) was dissolved in methanol (250 mL) at room temperature. Water (40 mL) was added followed by stannous chloride (5 g, 26.3 mmol). An additional amount (4 g, 21.0 mmol) of stannous chloride solution was added and stirring was continued for 1 hour at room temperature. The solution was stirred for an additional 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 (to 200 mL). The residual aqueous solution was extracted with ethyl acetate (2 × 200 mL). The ethyl acetate fractions were combined, dried over sodium sulfate (20 g), filtered and concentrated under reduced pressure. The clear residual foam was purified by filtration through silica gel (150 g) and eluting with dichloromethane / methanol (2: 1). From this procedure, glycine ester of cyclosporin A (8.54 g, 89% yield) was obtained as a transparent solid foam.
Glycine ester of cyclosporin A:

¹ H NMR (300 MHz, CDC 1 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, 53 H).

¹³ C NMR (75 MHz, CDC 1 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, 54.74, 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, 24.54, 24.31, 23.91, 23.78, 23.68, 21.86, 21. 33, 2 .25, 20.68, 19.76, 18.74, 18.24, 17.94, 17.79, 15.18, 10.03.

シクロスポリンＡのグリシンエステルＳＰＩ００１２０３（２．０ｇ、１．５９ミリモル）を、ｂｏｃ−Ｌ−リジン（１．３１ｇ、３．７８ミリモル）およびＥＤＣ（０．７５ｇ、３．９ミリモル）と共に、窒素雰囲気下、室温で無水ジクロロメタン（２５ｍＬ）に溶解した。ｂｏｃ−Ｌ−リジンを、冷えた硫化水素カリウム溶液（５０ｍＬの水に１ｇ）で抽出してからさらに冷水（２×５０ｍＬ）で抽出することによって、ジシクロロヘキシルアミン塩（５０ｍＬのエーテル中に２．０ｇ）から調製した。ｂｏｃ−Ｌ−リジンを含有するエーテルを、硫酸ナトリウム（５ｇ）を通して乾燥し、濾過、濃縮して、高真空下、１時間、室温で乾燥した。ＥＤＣ、ｂｏｃ−Ｌ−リジンおよびシクロスポリンＡのグリシンエステルの混合物に、いくつかのＤＭＡＰの結晶を加え、溶液を４時間、室温で攪拌し続けた。ジクロロメタン溶液をＤＩＵＦ水（５０ｍＬ）、５％重炭酸ナトリウム溶液（５０ｍＬ）、およびＤＩＵＦ水（５０ｍＬ）で抽出した。硫酸ナトリウム（１０ｇ）を通して乾燥した後、ジクロロメタン溶液を濾過し、減圧下で濃縮した。白色の残留発泡体（３．０１ｇ）を、シリカゲル（５０ｇ）を用い、ヘプタン／アセトン（２：１）で溶出するフラッシュカラムクロマトグラフィーにて精製した。留分含有生成物を組み合わせ、減圧下で濃縮し、高真空下に乾燥した。精製した保護中間体（白色固体２．３４ｇ、収率９２．８％）を、アルゴン雰囲気下のフラスコに入れ、これを氷水槽の中で冷却した。冷した無水の２Ｍ塩酸ジエチルエーテル（２０ｍＬ）溶液を加え、溶液を８時間（５℃で）攪拌した。混合物をゆっくり、一晩かけて室温に暖めた。合計２０時間攪拌した後、フラスコを再度氷水槽で３０分間冷却した。生成物を濾過し、高真空下、１時間、室温で乾燥し、さらに５０℃で４時間乾燥した。この実験から、白色の固体として、シクロスポリンＡのＮ−（ｌ−リジン）−グリシンエステル、二塩酸塩三水和物ＳＰＩ００２２（１５．９ｇ、収率７３．９％）を得た。

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣ１_３， ＮＭＲ ｄａｔａ ｉｓ ｆｏｒ ｔｈｅ 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， １７ Ｈ）， 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）．

^１３Ｃ ＮＭＲ （７５ ＭＨｚ， ＣＤＣ１_３， ＮＭＲ ｄａｔａ ｉｓ ｆｏｒ ｔｈｅ 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， ３２．４４， 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分析：
Ｃａｌｃｕｌａｔｅｄ ｆｏｒ Ｃ_７０Ｈ_１２８Ｃｌ_２Ｎ_１４Ｏ_１５−３Ｈ_２Ｏ ： Ｃ ５５．５０， Ｈ ８．９２， ａｎｄ Ｎ １２．７４ ； ｆｏｕｎｄ： Ｃ ５８．２８， Ｈ ８．９８， ａｎｄ Ｎ １３．１６．

ＨＰＬＣ分析：
９９．６０％ ｐｕｒｉｔｙ ； ｒ． ｔ． ＝ １４．７６３ ｍｉｎ．； ８０％ ａｃｅｔｏｎｉｔｒｉｌｅ／２０％ Ｔｒｉｓ ｂａｓｅ ｉｎ ＤＩＵＦ ｗａｔｅｒ； １ ｍＬ／ｍｉｎ ； ６０Ｃ； Ｓｙｎｅｒｇｉ Ｈｙｄｒｏ ＲＰ， ４ｕ ｃｏｌｕｍｎ （ｓｅｒｉａｌ ＃ １６３３８３−７）， ４．６ｘ２５０ ｍｍ ； ２０ ｕｌ； ＵＶ＝２１０ｎｍ．

融点： １９６．０−１９８ ℃ （ｕｎｃｏｒｒｅｃｔｅｄ） 4) SPI0022

Cyclosporin A glycine ester SPI001203 (2.0 g, 1.59 mmol) together with boc-L-lysine (1.31 g, 3.78 mmol) and EDC (0.75 g, 3.9 mmol) under nitrogen atmosphere , Dissolved in anhydrous dichloromethane (25 mL) at room temperature. Boc-L-lysine was extracted with cold potassium hydrogen sulfide solution (1 g in 50 mL water) and then with cold water (2 × 50 mL) to give dicyclohexylamine salt (2 in 50 mL ether). 0.0 g). The ether containing boc-L-lysine was dried over sodium sulfate (5 g), filtered, concentrated and dried under high vacuum for 1 hour at room temperature. To a mixture of EDC, boc-L-lysine and glycine ester of cyclosporin A, several crystals of DMAP were added and the solution was kept stirring at room temperature for 4 hours. The 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 white residual foam (3.01 g) was purified by flash column chromatography using silica gel (50 g) eluting with heptane / acetone (2: 1). The fraction-containing products were combined, concentrated under reduced pressure and dried under high vacuum. The purified protected intermediate (2.34 g of white solid, 92.8% yield) was placed in a flask under an argon atmosphere and cooled in an ice water bath. A cooled anhydrous 2M diethyl ether hydrochloride (20 mL) solution was added and the solution was stirred for 8 hours (at 5 ° C.). The mixture was slowly warmed to room temperature overnight. After stirring for a total of 20 hours, the flask was again cooled in an ice water bath for 30 minutes. The product was filtered, dried under high vacuum for 1 hour at room temperature, and further dried at 50 ° C. for 4 hours. From this experiment, N- (1-lysine) -glycine ester of cyclosporin A, dihydrochloride trihydrate SPI0022 (15.9 g, yield 73.9%) was obtained as a white solid.

¹ H NMR (300 MHz, CDC 1 ₃ , NMR data for the 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, 17 H), 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, 50H).

¹³ C NMR (75 MHz, CDC 1 ₃ , NMR data for the 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, 32.44, 31.83, 31.50, 30.38, 30.06, 29.81, 29.55, 25.14, 24.90, 24.52, 24.43, 24.00, 23. 76, 1.93, 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; r. t. = 14.763 min. 80% acetonitrile / 20% Tris base in DIUF water; 1 mL / min; 60C; Synergy Hydro RP, 4u column (serial # 163383-7), 4.6 × 250 mm;

Melting point: 196.0-198 ° C. (uncorrected)

シクロスポリンＡのグリシンエステル（ＳＰＩ００１２０３、７．５０ｇ、５．９５ミリモル）を、ｂｏｃ−Ｌ−プロリン（２．５６ｇ、１１．９０ミリモル）およびＥＤＣ（２．２８ｇ、１１．９０ミリモル）と共に、アルゴン雰囲気下、室温で無水ジクロロメタン（５０ｍＬ）に溶解した。ＥＤＣ、ｂｏｃ−Ｌ−プロリンおよびシクロスポリンＡのグリシンエステルの混合液に、ＤＭＡＰの結晶数個を加え、溶液を３時間、室温で攪拌し続けた。ジクロロメタン溶液をＤＩＵＦ水（５０ｍＬ）および５％重炭酸ナトリウム溶液（２ ｘ ５０ｍＬ ）で抽出した。硫酸ナトリウム（１０ｇ）上で乾燥した後、ジクロロメタン溶液を濾過し、減圧下に濃縮した。残った白色の泡沫（９．５０ｇ）を、シリカゲル（１５０ｇ）を用いて、ヘプタン／アセトン（２：１、続いて１：１）で溶出するフラッシュカラムクロマトグラフィーにて精製した。生成物含有分画を一つにまとめ、減圧下に濃縮し、高真空下で１０分間、室温で乾燥した（白色固体７．９４ｇ、収率９１．７％）。 5) SPI0023

Cyclosporine A glycine ester (SPI001203, 7.50 g, 5.95 mmol) together with boc-L-proline (2.56 g, 11.90 mmol) and EDC (2.28 g, 11.90 mmol) in an argon atmosphere The solution was dissolved in anhydrous dichloromethane (50 mL) at room temperature. To a mixture of EDC, boc-L-proline and glycine ester of cyclosporin A, several crystals of DMAP were added and the solution was kept stirring at room temperature for 3 hours. The dichloromethane solution was extracted with DIUF water (50 mL) and 5% sodium bicarbonate solution (2 × 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 column chromatography using silica gel (150 g) eluting with heptane / acetone (2: 1 followed by 1: 1). Product-containing fractions were combined, concentrated under reduced pressure, and dried under high vacuum for 10 minutes at room temperature (7.94 g, white solid, yield 91.7%).

The purified protected intermediate (6.46 g) was placed in a flask under an argon atmosphere and cooled in an ice water bath. A cooled anhydrous 2M diethyl ether solution (150 mL) was added and the solution was stirred for 8 hours (at 5 ° C.). The mixture was slowly warmed to room temperature overnight. After stirring for a total of 20 hours, the flask was again cooled in an ice water bath for 30 minutes. The product was filtered and dried at room temperature under high vacuum for 30 minutes. N- (L-proline) -glycine ester hydrochloride of cyclosporin A (5.17 g, yield 84.6%, purity 90% by HPLC) was added to DIUF water (25 mL) containing sodium bicarbonate (1 g). And converted to the free base. The free base was extracted with dichloromethane (3 × 25 mL), which was dried over sodium sulfate (5 g), filtered and concentrated. The remaining grayish white solid (5 g) was purified by filtration through silica gel (100 g), eluting with acetone. Product containing fractions were combined, concentrated under reduced pressure and dried under high vacuum at room temperature for 30 minutes. While cooling in an ice bath, the free base (3.8 g) was dissolved in diethyl ether (25 mL) and added to anhydrous 2M hydrochloric acid (5 mL) heptane (50 mL) solution to regenerate the hydrochloride salt. After 20 minutes at 5 ° C., the white solid was filtered and dried under high vacuum for 6 hours at room temperature. From this experiment, N- (1-proline) -glycine ester hydrochloride (SPI0023, 3.8 g) of cyclosporin A was obtained as a white solid.

¹ H NMR (300 MHz, CDC 1 ₃ ):
δ = 14.20 (br s, 2H), 8.62 (d, 1 H, 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), 78-1.20 (m, 15H), 1.15-0.66 (m, 46H).

¹³ C NMR (75 MHz, CDC 1 ₃ ):
δ = 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, 57.36, 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, 30.43, 29.93, 29.72, 29.30, 29.16, 27.56, 26. 04, 5.00, 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} O 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; r. t. = 10.613 min. 85% acetonitrile / 15% Tris base in DIUF water; 1.2 mL / min; 60C; Synergy Hydro RP, 4u column (serial # 163383-7), 4.6 × 250 mm;

Melting point: 197.0-199 ° C. (uncorrected)

These cyclosporine prodrugs of the present invention are effective in the treatment of diseases or conditions for which macrocyclic immunosuppressants are generally used. These prodrugs are converted in the body to release the active compound, thereby treating macrocyclic immunosuppressive agents by reducing or eliminating the biopharmaceutical and pharmacokinetic disturbances associated with each macrocyclic immunosuppressive agent. Increase benefits. However, it is also noted that these prodrugs themselves have sufficient activity in mammals without releasing the active agent. Prodrugs are more water soluble than cyclosporine or other macrocyclic immunosuppressants, and must be accompanied by a carrier vehicle that can be toxic or cause unwanted side effects, such as alcohol or castor oil. Absent. Furthermore, oral preparations containing prodrugs are absorbed by the blood and are extremely effective.

Thus, the prodrugs of cyclosporine of the present invention enhance therapeutic benefit by eliminating biopharmaceutical and pharmacokinetic obstacles.

Furthermore, the prodrug is easily synthesized in high yield using commercially available reagents.

VI. Valproic acid ester valproic acid (2-propylpentanoic acid) is a low molecular weight carboxylic acid derivative and is widely used as an anticonvulsant and is useful for the treatment of epilepsy. In addition, it has vasodilator activity in the brain, reducing migraine. Valproic acid (2-propylpentanoic acid) is administered orally to suppress epilepsy symptoms in humans and relieve severe pain associated with migraine.

Valproic acid is known to have many therapeutic applications, which are extremely diverse and somewhat surprising. For example, in addition to its efficacy in treating epilepsy and migraine, it is effective in treating certain types of psychosis such as bipolar disorder, mood stabilization, aggression suppression, personality disorder impulses, and dementia agitation And has also been used as an adjuvant treatment for the treatment of post-traumatic stress disorder (PTSD).

Mechanism of action:
Despite its long use in the treatment of epilepsy, the exact mechanism of action of valproic acid is not yet known. It is hypothesized that its effect is exerted by increasing the concentration of gamma-aminobutyric acid (GABA) in the brain. Gamma-aminobutyric acid is a chemical, neurotransmitter, that nerves use to communicate with each other.

Valproate is a selective drug for myoclonic epilepsy with or without generalized tonic-clonic seizures, including Janz's juvenile myoclonic epilepsy, which occurs in adolescents and young adults. Photosensitive myoclonus is usually easy to suppress. Valproate is effective in the treatment of benign myoclonic epilepsy and post-hypoxic myoclonus, and is also used with clonazepam in the treatment of severe progressive myoclonic epilepsy characterized by tonic-clonic seizures. It can also be used for certain types of stimulus-sensitive epilepsy (reflective epilepsy, startle epilepsy).

Valproate is also effective in childhood spasms, but is relatively contraindicated in children whose spasm is caused by hyperglycinemia or other metabolic (mitochondrial) abnormal underlying diseases. In general, atonic and ataxic seizures in patients with Lennox-Gastaut syndrome are difficult to suppress, but valproate is a treatment of choice for mixed seizures. Because this drug is useful in some patients who are refractory to all other antiepileptic drugs, it may be tried in almost all non-responsive patients, regardless of the type of seizure.

Despite its usefulness, hepatotoxicity can be fatal and specific and cannot be prevented by normal liver enzyme monitoring. Hepatotoxicity occurs in very small children and is most common when using multiple anticonvulsants. Valproate-induced cytopenia is dose-dependent and requires monitoring of the total blood count during treatment. Encephalopathy with hyperammonemia that does not show abnormal liver function tests may occur. In the first month of pregnancy, there is a risk of neural tube defects.

Valproic acid is a low molecular weight liquid with a specific odor. When taken orally, it tastes unpleasant and strongly stimulates the mouth and throat. In order to convert valproic acid into a solid dosage form convenient for oral administration, several derivatives of covalently and ionically bound carboxylic acids have been made. A simple sodium salt of valproic acid, which is sodium valproate, is available as a solid. However, a stable coordination complex, known as divalproex sodium, was formed by partially neutralizing two molecules of valproic acid with one atom of sodium. This product is the most widely available valproic acid half salt and is sold by Abbott Laboratories, USA under the trade name Depakote®. Depakote® also has sustained release formulations for oral administration.

A major drawback of valproic acid is that it is difficult to administer when in liquid form. Furthermore, administration of valproic acid in different forms does not exhibit the desired bioavailability uniformly. For example, the total bioavailability of valproate from valproic acid, its sodium salt, divalprox (Divalproex®) and their sustained release formulations is not interchangeable. Because continuous monitoring of the plasma profile of valproic acid is essential, changes in plasma concentration due to formulation changes adversely affect overall treatment outcome.

Prodrugs of valproic acid that overcome some of the difficulties described above to improve therapeutic efficacy, homogenize blood profiles, develop pharmaceutically sophisticated formulations, and reduce first pass metabolism Describe.

To date, there are no commercially available pharmaceutical preparations that can deliver valproic acid without harmful side effects. However, the present invention provides a number of water-soluble and non-toxic valproic acid derivatives that are suitable for stable delivery of valproic acid in the body without harmful side effects and without the need for expensive additives and excipients. Produced.

Accordingly, in one aspect, the present invention is directed to prodrugs of valproic acid. A prodrug consists of the hydroxyl group of an amino acid esterified to a free carboxyl group present on the valproic acid molecule. In another aspect, the amine group of the amino acid reacts with the COOH group to form an amide bond.

または、その薬学的に許容される塩を対象とする。式中のＲはＮＨ−ＡＡまたはＯ−ＡＡであり、ＡＡはアミノ酸であって、アミン基またはヒドロキシル基のいずれかがバルプロ酸のカルボン酸基と反応する。 More specifically, embodiments of the present invention include compounds of the formula

Alternatively, a pharmaceutically acceptable salt thereof is targeted. R in the formula is NH-AA or O-AA, and AA is an amino acid, and either an amine group or a hydroxyl group reacts with a carboxylic acid group of valproic acid.

The present invention is also directed to a pharmaceutical composition comprising a therapeutically effective amount of the various valproic acid prodrugs described above and a pharmaceutical carrier therefor.

In another aspect, the invention is directed to a method of treating a patient in need of valproic acid treatment, the method of treatment comprising administering to the patient an effective amount of valproic acid.

In a further aspect, the present invention converts liquid valproic acid to a solid powder by reacting the carboxyl functionality of valproic acid with the amine functionality or hydroxyl functionality of an amino acid and isolating the product. Target method.

In yet another aspect, the present invention is directed to a method of reducing or eliminating potential first-pass metabolism in a substantially and therapeutically effective manner, thereby administering a prodrug to a patient. Improve the stability of the therapeutic effect. The method involves reacting the COOH functionality of a valproic acid molecule with the NH2 functionality or OH functionality of a selected amino acid, forming an ester covalent bond or an amide covalent bond, respectively, and isolating the product. And administering a prodrug to the patient.

When an unsubstituted natural amino acid is esterified to valproic acid, the resulting prodrug becomes a pharmaceutically refined free-flowing powder that is rapidly absorbed by the body and cleaved in the body to remove non-toxic amino acids. It has been discovered that it does not require release as well as emulsifiers, additives and other excipients.

Furthermore, it has been found that the present invention also produces drugs, which are prodrugs of valproic acid, are highly effective antiepileptic agents, and show their effects in a complete form. Thus, the amino acid prodrugs are effective antiepileptic agents, are useful in the treatment of many psychoses, and exhibit such efficacy whether or not they release the active parent drug.

Because the bulk density of prodrugs of parproic acid is higher than the corresponding sodium salt, they are suitable for miniaturizing large weight tablets and capsules. Furthermore, the prodrug of palproic acid is free from the bitterness of valproic acid and the unpleasant odor.

The prodrug of the present invention is considered to have no acid activity because the carboxyl group that causes acid activity is blocked. Regardless of whether the prodrug releases valproic acid or not. It has been shown to be an effective antiepileptic agent. However, all described valproic acid prodrugs release active agents with all pharmacological and psychoactive properties in vivo.

Prodrugs of valproic acid clearly offer many advantages over valproic acid. For example, the side chains cleaved from these prodrugs are all natural essential amino acids and are therefore non-toxic. As a result, it shows a high therapeutic index. Second, all prodrugs are easily cleaved in the body to release valproic acid. Furthermore, because it is highly water soluble, it can be used to make in situ solutions just prior to IV administration using sterile lyophilized powder, or to provide drugs in the form of syringes or bottles filled with medicinal solutions for injection. By doing so, it can be administered easily. Amino acid esters are more stable than valproic acid because the COOH group in valproic acid is blocked from reaction with the base. As a result, the prodrugs of valproic acid of the present invention are more effective than valproic acid itself, without the toxicity and other pharmaceutical problems associated with currently marketed formulations.

An overview of the synthetic procedure for L-serine, L-threonine and L-hydroxyproline esters of valproic acid (2-propylpentanoic acid) is given in the Synthetic Sequence section, which is illustrative for the preparation of various prodrugs of the present invention. . Complete procedures and analytical data are given in the experimental section. In general, valproic acid (2-8 g, batch) was coupled with N-benzyloxy / benzyl ester protected amino acid using EDC in the presence of a catalytic amount of DMAP. After completion of the reaction (20 hours at room temperature), the mixture was extracted with DIUF water, dried over sodium sulfate, and concentrated under reduced pressure. The crude material was used directly in the deprotection step or purified by column chromatography. From the procedure, protected amino acid esters of valproic acid were obtained in yields ranging from 72% to 92%. The protecting group was removed by hydrogenation (30 psi H ₂ ) in the presence of 10% palladium-carbon. The amino acid ester of valproic acid was extracted using ethanol to remove the palladium catalyst, concentrated, and dried. Finally, it was acidified with hydrochloric acid to make a salt. The crude salt (57% to 92% yield) was then purified by the method described in the experimental section.

バルプロ酸のＬ−セリン、Ｌ−スレオニンおよびＬ−ヒドロキシプロリンエステルの合成：ａ）ＥＤＣ、ＤＭＡＰ、ＣＨ_２Ｃｌ_２；ｂ）Ｈ_２、１０％Ｐｄ／Ｃ、ＥｔＯＨ、ＥｔＯＡｃ；ｃ）ＨＣｌ Synthesis order:

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) HCl

Experiment chapter:
SPIC001, SPIC002 and SPIC003 were synthesized in 1 or 2 batches. Reagents listed in the experimental section were purchased from Lancaster, Sigma-Aldrich, Acros or Bachem with the highest purity available, except for solvents purchased from Fischer Scientific or Mallinckrodt.

^１Ｈ 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）．

^１３Ｃ 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． 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 mmol), N-carbobenzyloxy-L-serine benzyl ester (Z-Ser-OBzl, 14.80 g, 44.93 mmol), EDC (8 .61 g, 44.91 mmol) and DMAP (549 mg, 4.49 mmol) in anhydrous dichloromethane (50 mL) were stirred at room temperature for 20 hours under an argon atmosphere. After 20 hours, the dichloromethane was washed with water (3 × 50 mL), dried over magnesium sulfate (5 g), filtered and concentrated under reduced pressure. The remaining colorless oil (20.87 g) was purified by column chromatography using silica gel (150 g, 0.035 to 0.070 mm, pore size 6 nm) eluting with hexanes / ethyl acetate (3: 1). The product-containing fractions were concentrated under reduced pressure and dried under high vacuum until the weight was constant. From experiments, the protected L-serine-valproic acid ester SPIC00101 (18.9 g, yield 92) was obtained as a colorless oil. %).

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.

^１Ｈ 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）．

^１３Ｃ 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分析：
９８．４９％ ｐｕｒｉｔｙ； ｒｔ＝ ４．７６７ ｍｉｎ； Ｌｕｎａ Ｃ１８ ５ｕ ｃｏｌｕｍｎ （ｓｎ １６７９１７−１３）； ４．６ｘ２５０ ｍｍ； ２５４ ｎｍ； ３３％ ＡＣＮ／６６％ ＤＩＵＦ ｗａｔｅｒ； ３５ Ｃ； ２０ ｕｌ ｉｎｊ． ； １ ｍｌ／ｍｉｎ ； ２０ ｍｇ／ｍＬ ｓａｍｐｌｅ ｓｉｚｅ； ｓａｍｐｌｅ ｄｉｓｓｏｌｖｅｄ ｉｎ ｍｏｂｉｌｅ ｐｈａｓｅ．

ＣＨＮ分析：
ｃａｌｃ．： Ｃ ４９．３４， Ｈ ８．２８， Ｎ ５．２３ ； ｆｏｕｎｄ： Ｃ ４９．２２， Ｈ ８．３５， Ｎ ５．２４．

融点： １５９−１６０ ℃ Protected L-serine-valproic acid ester SPIC00101 (18.9 g, 41.48 mmol) was dissolved in ethanol (60 mL) and ethyl acetate (60 mL) at room temperature and 10% palladium-carbon (3.0 g) under nitrogen atmosphere. , 50% wet rate) in a Parr bottle (500 mL). The nitrogen atmosphere was exchanged with hydrogen gas (30 psi). After shaking for 4 hours, a solution of palladium catalyst (1.0 g) in ethanol / ethyl acetate (1: 1, 100 mL) was added and the reaction mixture was shaken overnight at room temperature under hydrogen gas (30 psi). After 24 hours, the catalyst was filtered through a thin layer of activated carbon. Ethanol and ethyl acetate were concentrated at room temperature under reduced pressure. After drying under high vacuum, the remaining solid was acidified with hydrochloric acid in diethyl ether (2M, 24.6 mL). The mixture was stored in the refrigerator for 2 hours, then filtered and washed with additional cold diethyl ether (10 mL). After filtration, the remaining white solid was dried under high vacuum at room temperature until the weight was constant (24 hours). From the experiment, L-serine-valproate ester / hydrochloride SPIC001 (6.34 g, 57% yield) was obtained as a white solid.

¹ 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:
Rt = 4.767 min; Luna C18 5u column (sn 167917-13); 4.6 × 250 mm; 254 nm; 33% ACN / 66% DIUF water; 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 ° C

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＣＤＣｌ_３）： δ ＝ ７．２９ （１０Ｈ， 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）．

^１３Ｃ 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， ７１．５３， 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． 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 mmol), N-carbobenzyloxy-L-hydroxyproline benzyl ester (Z-Hyp-OBzl, 10.66 g, 30 mmol) ¹ , EDC (5.74 g) , 30 mmol) and DMAP (366 mg, 3 mmol) in anhydrous dichloromethane (30 mL) were stirred at room temperature under an argon atmosphere for 20 hours. After 20 hours, the dichloromethane was washed with water (3 × 30 mL), dried over magnesium sulfate (5 g), filtered and concentrated under reduced pressure. The remaining colorless oil SPIC00201 (11.95 g, 24.7 mmol, yield 82.4%) was used as such without purification.

¹ H NMR (300 MHz, CDCl ₃ ): δ = 7.29 (10H, 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, 71.53, 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 mmol) was dissolved in ethanol (50 mL) and ethyl acetate (100 mL) at room temperature and then 10% palladium-carbon (3. 5 g, 50% wet rate) in a Parr bottle (500 mL). The nitrogen atmosphere was exchanged with hydrogen gas (30 psi). After shaking for 15 hours, the catalyst was filtered off through a thin layer of celite and activated carbon. The ethanol and ethyl acetate mixture was concentrated under reduced pressure at room temperature. When dried overnight at room temperature under high vacuum, L-hydroxyproline-valproic acid ester SPIC002 (9.2 g, 99.8% yield) was obtained as a white solid from the experiment. To remove trace amounts of impurities, zwitterions were purified in two batches using reverse phase column chromatography (50 g ODS silica gel). Zwitterions were applied to a column equilibrated with DIUF water and eluted with a mixture of DIUF water / methanol (2: 1, 1: 1, 1: 2, 100% methanol).

^１Ｈ 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＝ １０．５ 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）．

^１３Ｃ 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分析：
９９．２０％ ｐｕｒｉｔｙ ； ｒ． ｔ． ＝ 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分析：
ｃａｌｃ．： Ｃ ６０．６８， Ｈ ９．０１， Ｎ ５．４４ ； ｆｏｕｎｄ： Ｃ ６０．５８， Ｈ ９．１２， Ｎ ５．４８．

融点： １７９．０−１８０．０ ℃ The product-containing fractions were combined and concentrated under reduced pressure at 20 ° C. (below) and then dried at room temperature under high vacuum until the weight was constant (24 hours, recovering 6.4 g of white solid) ).

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 (brd, 1H, J = 10.5 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, CDC 1 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; r. t. = 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;

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 ° C

^１Ｈ 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）．

^１３Ｃ 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． 3) SPIC003: 2-propyl-pentanoic acid 2 (S) -amino-2-carboxy-1 (R) -methyl-ethyl ester / hydrochloride (L-threonine-valproic acid ester / hydrochloride)
2-propylpentanoic acid (valproic acid, 4.32 g, 30 mmol), N-carbobenzyloxy-L-threonine benzyl ester (Z-Thr-OBzl, 10.30 g, 30 mmol), EDC (5.74 g, 30 mmol) Mmol) and DMAP (366 mg, 3.0 mmol) in anhydrous dichloromethane (30 mL) were stirred at room temperature for 20 hours under an argon atmosphere. After 20 hours, the dichloromethane was washed with water (3 × 30 mL), dried over magnesium sulfate (5 g), filtered and concentrated under reduced pressure. The remaining colorless oil (13.44 g) was purified by column chromatography using silica gel (100 g, 0.035 to 0.070 mm, pore size 6 nm) eluting with hexanes / ethyl acetate (4: 1). The product-containing fractions were concentrated under reduced pressure and dried under high vacuum until the weight was constant. From experiments, the protected L-threonine-valproic acid ester SPIC00301 (12.65 g, yield 89.89) was obtained as a colorless oil. 8%).

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-valproic acid ester SPIC00301 (12.65 g, 26.9 mmol) was dissolved in ethanol (50 mL) and ethyl acetate (50 mL) at room temperature and 10% palladium-carbon (2.53 g) under nitrogen atmosphere. , 50% wet rate) in a Parr bottle (500 mL). The nitrogen atmosphere was exchanged with hydrogen gas (30 psi). After 20 hours, the catalyst was filtered through a thin layer of activated carbon and washed away with ethanol (25 mL). Ethanol and ethyl acetate were concentrated at room temperature under reduced pressure. After drying under high vacuum, the remaining solid (6.13 g) was acidified with a DIUF aqueous solution (50 mL) of hydrochloric acid (3.1 mL concentrated hydrochloric acid). The solution was subjected to a second filtration through activated charcoal and dried overnight in a lyophilizer. From the experiment, L-threonine-valproic acid ester / hydrochloride SPIC003 (6.52 g, yield 86.0%) was obtained as a white solid.

^１Ｈ ＮＭＲ （３００ ＭＨｚ， ＤＭＳＯ）： δ ＝ ８．７１ （ｂｒ ｓ， ３Ｈ）， ５．２８ （ｍ， １Ｈ）， ４．１６ （ｄ， １Ｈ， Ｊ＝ ２．７Ｈｚ）， ２．３３ （ｍ， １Ｈ）， １．５６−１．４０ （ｍ， ２ Ｈ）， １．３７−１．２７ （ｍ， ５Ｈ）， １．２１−１．１３ （ｍ， ４Ｈ）， ０．８４ （ｔ， ６Ｈ， Ｊ＝ ６．６ Ｈｚ）．

^１３Ｃ 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分析：
９８．８８％ ｐｕｒｉｔｙ； ｒ． ｔ． ＝ ４．８６４ ｍｉｎ．； ７０％ ＤＩＵＦ ｗａｔｅｒ／３０％ ａｃｅｔｏｎｉｔｒｉｌｅ ； １ ｍＬ／ｍｉｎ； ４０Ｃ； Ｌｕｎａ Ｃ１８， ５ｕ ｃｏｌｕｍｎ （ｓｅｒｉａｌ ＃ ２１１７３９−４２）， ４．６ｘ２５０ ｍｍ； ２０ ｕｌ ｉｎｊｅｃｔｉｏｎ； ｓａｍｐｌｅ ｄｉｓｓｏｌｖｅｄ ｉｎ ｍｏｂｉｌｅ ｐｈａｓｅ．

ＣＨＮ分析：
ｃａｌｃ．： Ｃ ５１．１５， Ｈ ８．５９， Ｎ ４．９７ ； ｆｏｕｎｄ： Ｃ ５１．２９， Ｈ ８．５９， Ｎ ４．９８．

融点： １４４ ℃ A batch of L-threonine-valproate ester hydrochloride SPIC003 (8.8 g) combined was purified by crystallization from acetonitrile. After the salt was dissolved in warm acetonitrile (225 mL), the material was treated with activated carbon, filtered and placed in a 5 ° C. refrigerator overnight. After 18 hours, the white solid was filtered, washed with cold acetonitrile (10 mL) and then dried at room temperature under high vacuum until the product weight was constant (24 hours). From this step, L-threonine-valproic acid ester / hydrochloride SPIC003 (6.82 g, yield 77.5%) was recovered 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; r. t. = 4.864 min. 70% DIUF water / 30% acetonitrile; 1 mL / min; 40C; Luna C18, 5u column (serial # 211739-42), 4.6x250 mm; 20 ul insole sorbed sample;

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 ° C

The solubility of the ester was determined by precipitating suspended matter for several hours after dissolving an excess amount of each drug in water at room temperature. The resulting solution was centrifuged at 1500 rpm for 3 minutes, and the supernatant was analyzed. These esters were found to have a solubility in water exceeding 50 mg / mL.

There are a number of screening tests that determine the usefulness of prodrugs made according to the disclosed methods. Such methods also include in vitro and in vivo screening methods.

In vitro methods include acid / base hydrolysis of prodrugs, hydrolysis in porcine pancreas, hydrolysis in rat intestinal fluid, hydrolysis in human gastric fluid, hydrolysis in human intestinal fluid and hydrolysis in human plasma Is included. These assays are described in Simmons, DM, Chandran, VR and Portmann, GA, Danazol, Amino Acid Prodrugs, In Vitro and In Situ Biopharmaceutical Evaluation, Drug Development. The entire contents of which are incorporated by reference.

The prodrugs of valproic acid of the present invention are effective in treating diseases or conditions for which valproic acid is usually used. The prodrugs disclosed herein are converted in the body to release the active compound, thereby reducing the therapeutic benefit of valproic acid by reducing or eliminating the biopharmaceutical and pharmacokinetic disorders associated with valproic acid. Strengthen. However, it should be noted that these prodrugs themselves have sufficient activity in mammals without releasing the active agent.

Thus, the prodrugs of the present invention enhance the therapeutic benefit by removing the biopharmaceutical and pharmacokinetic obstacles of existing drugs.
Furthermore, prodrugs are easily synthesized in high yields using commercially available reagents that can be purchased easily.

VII Water-soluble prodrugs of fibric acid derivatives Fibric acid derivatives are useful for the treatment of hyperlipidemia in mammals with symptoms of high triglycerides, low HDL (high density lipoprotein or “good” cholesterol), and high cholesterol. It is a useful antihyperlipidemic drug. Fibric acid derivatives are also useful in reducing LDL (low density lipoprotein, or “bad” cholesterol). The general structure of the fibric acid analog is shown below, where X is various mixed aliphatic and aromatic functional groups. Specific derivatives included in this formula are clofibric acid, fenofibric acid, ciprofibrate, gemfibrozil and the like.

A typical example of the chemical moiety X in the above structure is shown below.

The fibric acid analogs represented by the above structural formula have been found to be applicable to many treatments, and the therapeutic application examples are extremely diverse and a little surprising. These derivatives are widely useful for the treatment of dyslipidemia and dyslipoproteinemia. In the present specification, the definitions of dyslipidemia and dyslipoproteinemia include hypercholesterolemia, abnormal value and high value of cholesterol, abnormal value and high value of LDL cholesterol, abnormal value and high value of total cholesterol Abnormalities and highs of plasma cholesterol, abnormalities and highs of triglycerides, hypertriglyceridemia, abnormalities of lipoproteins, abnormalities and highs of low density lipoprotein (LDL), abnormalities and highs of very low density lipoprotein Abnormalities and highs of very low density / intermediate density lipoproteins, abnormalities of high density lipoproteins, hyperlipidemia, hyperchylomicronemia, abnormalities of chylomicrons, related disorders, and combinations thereof. These include: ILIB Lipid Handbook for Clinical Practice, Blood Lipids and Coronary Heart Disease, Second Edition, A.M. M.M. Goto et al. , International Lipid Information Bureau, New York, N .; Y. , 2000, incorporated herein by reference.

Mechanism of action:
The mechanism of action of fibric acid derivatives found in clinical practice is in vivo by transgenic mice and in vitro in human hepatocyte cultures by activation of peroxisome proliferator-activated receptor alpha (PPAR-alpha). Explained. Through this mechanism, fibric acid derivatives increase lipolysis by activating lipoprotein lipase and at the same time reducing the production of apoprotein C-III (inhibitor of lipoprotein lipase activity), and particles rich in triglycerides from plasma Eliminate.

The resulting reduction in triglycerides results in changes in the size and composition of LDL, from small, dense particles (which are thought to be atherogenic because they are susceptible to oxidation) to large, floating particles Changes to. Larger particles have a higher affinity for cholesterol receptors and are rapidly degraded. Activation of PPAR-alpha also induces increased synthesis of apoproteins AI, A-II and HDL cholesterol.

Fibric acid derivatives are useful for the treatment of gout because they lower serum uric acid levels in hyperuricemia patients.

Types of hyperlipidemia include type I, type IIa, type IIb, type III, type IV and type V. These types can be characterized according to the level of lipids (cholesterol and triglycerides) and lipoproteins above normal values. Another classification is described in Drug Facts and Comarisons, 52nd Edition (1988) page 1066, which is incorporated herein by reference.

Many fibric acid derivatives do not provide sufficient bioavailability when administered orally, absorption is variable and unstable, and is food dependent. In fact, the absolute bioavailability of many fibric acid derivatives is not possible because the fibric acid prodrugs currently on the market are insoluble in water and parenteral formulations are difficult or unavailable. In addition, these drugs are not complete prodrugs because they are usually administered in ester form. These prodrugs need to be metabolized in the body to release the active drug, which is fibric acid. However, these drugs are extremely insoluble in water due to their ester formation, and are therefore difficult to formulate and are not easily degraded in the body for active drug release.

Many fibric acid derivatives are low to medium molecular weight solids with a characteristic odor. When taken orally, it tastes unpleasant and may strongly irritate the mouth and throat. When taken with food, blood levels are higher than when fasting. This bioavailability fed / fasted difference is more pronounced when fibric acid derivatives are compared to the corresponding prodrug derivatives. The overall bioavailability has been reported to be between 40 and 60, but varies greatly between patients.

One of the important problems associated with currently marketed fibric acid derivatives is that when these prodrugs are cleaved in the body, they release the prodrug moiety but are themselves highly toxic. For example, in the case of fenofibrate and gemfibrozil, isopropyl alcohol is released when the esterase enzyme cleaves the promoiety from fenofibric acid. It is well known that isopropanol is highly toxic when released into mammalian tissues.

In order to enhance the therapeutic effect, homogenize blood profile, develop pharmaceutically sophisticated formulations and improve the water solubility of the drug, the present invention replaces fibric acid derivatives that overcome many of the above difficulties. Discuss drugs.

Accordingly, in one aspect, the invention is directed to another class of prodrugs of fibric acid derivatives. Prodrugs consist of the hydroxyl group of an amino acid esterified to a free carboxyl group present on the fibric acid derivative molecule. In another embodiment, the amine group of the amino acid is reacted with COOH of fibric acid to form an amino bond.

（式中のｘは上記定義の通りであり）
または、その薬学的に許容される塩を対象とし、式中のＲはＮＨ−ＡＡまたはＯ−ＡＡであり、ＡＡはアミノ酸であって、このアミノ酸中のアミン基またはヒドロキシル基のいずれかをフィブリン酸誘導体のカルボン酸基と反応させる。 More specifically, in one embodiment of the present invention, a compound of the formula

(X in the formula is as defined above)
Or a pharmaceutically acceptable salt thereof, wherein R is NH-AA or O-AA, AA is an amino acid, and either an amine group or a hydroxyl group in the amino acid is fibrin; React with the carboxylic acid group of the acid derivative.

The present invention is also directed, in one aspect, to a pharmaceutical composition comprising a therapeutically effective amount of the various fibric acid derivative prodrugs described above and a pharmaceutical carrier therefor.

In another aspect, the invention is directed to a method of treating a patient in need of fibric acid derivative treatment, the method comprising administering to the patient a therapeutically effective amount of a fibric acid derivative.

In yet another aspect, the present invention relates to the conversion of a liquid fibric acid derivative into a solid powder by reacting the carboxyl functionality of the fibric acid derivative with the amine functionality or hydroxyl functionality of an amino acid and isolating the product. Target the method of conversion.

In yet another aspect, the present invention is directed to a method that allows a derivative to be readily absorbed by oral administration in a substantially and therapeutically effective manner, thereby stabilizing treatment by administering a prodrug to the patient. Improve the effect. The method comprises reacting the COOH functionality of a fibric acid derivative molecule with either the NH ₂ or OH functionality of a selected amino acid to form an ester or amide covalent bond, respectively, isolating the product, And administering the product to a patient.

When an unsubstituted natural amino acid is esterified to a fibric acid derivative, the resulting prodrug is a pharmaceutically refined and fluid powder, is absorbed quickly into the body, and is cleaved in the body to remove non-toxic amino acids. It has become certain that it does not release and does not require emulsifiers, additives and other excipients.

Furthermore, although the present invention also produces drugs, they are prodrugs of fibric acid derivatives, which are highly effective antihyperlipidemic drugs and show their effects in their full form. Thus, the amino acid prodrug is an effective antihyperlipidemic agent, useful for the treatment of many high cholesterol-related diseases, and whether such active parent drug is released or not. Show the effect.

Although the fibric acid prodrug of the present invention is considered not to have any acid activity because the carboxylic acid group responsible for acid activity is blocked, does the fibric acid prodrug release fibric acid derivative release? Regardless, it has been shown to be an effective antihyperlipidemic agent. However, all of the described fibric acid derivative prodrugs release active agents that have all of their pharmacological cholesterol-lowering properties in vivo.

The present invention clearly provides many advantages over fibric acid derivatives, for example, the side chains cleaved from these prodrugs are all naturally essential amino acids and are therefore non-toxic. This gives a high therapeutic index. Second, all prodrugs are easily cleaved in the body to release the fibric acid derivative. Furthermore, because of their high water solubility, they can be used to lyse the drug in lyophilized powder, by making an in situ solution just prior to IV administration, or in solution in advance in a syringe or bottle for injection. By providing a filled one, it can be easily administered. Amino acid esters are more stable than fibric acid derivatives because the COOH group of the fibric acid derivative is blocked from reaction with the base. Accordingly, the fibric acid derivative prodrugs described herein are more effective than fibric acid derivatives without the toxicity and other pharmaceutical problems associated with currently marketed formulations.

The prodrug of the present invention is a therapeutic agent for hyperlipidemia useful for the treatment of hyperlipidemia in mammals, whose symptoms are high triglycerides, low HDL (high density lipoprotein or “good cholesterol”) and high cholesterol. is there. Fibric acid derivatives are also useful in reducing LDL (low density lipoprotein, or “bad” cholesterol).

Representative examples of the synthesis of L-threonine, L-hydroxyproline and L-serine esters of fibric acid derivatives are shown schematically in the following synthetic steps. These procedures are equally applicable to all other compounds belonging to the fibric acid derivative class.

Synthesis of Fibric Acid Derivative Prodrugs The procedure for synthesizing L-threonine, L-hydroxyproline and L-serine esters of fibric acid derivatives is outlined in the synthetic order section, which is exemplary. Complete procedures and analytical data are given in the experimental section. In general, fenofibric acid (100 g batch) was prepared from 4-chloro-4′-hydroxybenzophenone according to literature procedures. Fenofibric acid was coupled with the t-butyl ester of N-Boc protected amino acids (L-serine, L-threonine and L-hydroxyproline) using EDC as a coupling agent and a catalytic amount of DMAP. The protecting group was removed with a mixture of hydrochloric acid in acetic acid (1M) and dichloromethane at low temperature (5 ° C., 3-6 days). The amino acid ester of fenofibric acid was purified by crystallization from ethyl acetate and dried under high vacuum.

フィブリン酸のＬ−セリン、Ｌ−スレオニンおよびＬ−ヒドロキシプロリンエステルの合成：ａ）Ｂｏｃ−Ｓｅｒ−ＯｔＢｕ、ＥＤＣ、ＤＭＡＰ、ＣＨ_２Ｃｌ_２；ｂ） Ｂｏｃ−Ｔｈｒ−ＯｔＢｕ、ＥＤＣ、ＤＭＡＰ、ＣＨ_２Ｃｌ_２；ｃ）Ｂｏｃ−Ｈｙｐ−ＯｔＢｕ、ＥＤＣ、ＤＭＡＰ、ＣＨ_２Ｃｌ_２；ｄ） ＨＣｌ、ＡｃＯＨ、ＣＨ_２Ｃｌ_２ Synthesis order:

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

Experiment chapter:
The synthesis of SPIB00201, SPIB00202 and SPIB00203 was performed in one or two batches. Reagents listed in the experimental section were purchased from Lancaster, Sigma-Aldrich, Acros or Bachem with the highest purity available, except for solvents purchased from Fischer Scientific or Mallinckrodt.

４−クロロ−４’−ヒドロキシベンゾフェノン（１１６ｇ、０．５００ｍｏｌｅ）および水酸化ナトリウム（１２０ｇ、３．００ｍｏｌｅ）のアセトン（１Ｌ）混合液を加熱して、２時間還流した。加熱を止め、熱源を取り外した。クロロホルム（１７９ｇ、１．５０ｍｏｌｅ）のアセトン（３００ｍＬ）溶液を、滴下して加えた。反応混合液を一晩、加熱せずに攪拌した。混合液を加熱しながら８時間還流し、次に室温まで冷やした。濾過して沈殿を取り除き、アセトン（１００ｍＬ）で洗った。濾過液を減圧下に濃縮して、茶色のオイルを得た。水（２００ｍＬ）を茶色のオイルに加え、１Ｎの塩酸で（ｐＨ＝１まで）酸性化した。形成した沈殿を濾過し、高真空下に乾燥した。残った黄色の固体（２６８ｇ）を４バッチに分けてトルエン（それぞれ４００ｍＬトルエン）から再結晶化した。濾過して高真空下に乾燥したところ、実験から淡黄色の固体として、フェノフィブリン酸（１１６ｇ、収率７３％）を得た。

^１Ｈ 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）．

^１３Ｃ 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． 1) Synthesis of fenofibric acid:

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. Heating was stopped and the heat source was removed. A solution 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 cooled to room temperature. The precipitate was removed by filtration and washed with acetone (100 mL). The filtrate was concentrated under reduced pressure to give a brown oil. Water (200 mL) was added to the brown oil and acidified with 1N hydrochloric acid (until pH = 1). The formed precipitate was filtered and dried under high vacuum. The remaining yellow solid (268 g) was recrystallized from toluene (400 mL toluene each) in 4 batches. Filtration and drying under high vacuum yielded fenofibric acid (116 g, 73% yield) as a pale yellow solid from the experiment.

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.

^１Ｈ 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）．

^１３Ｃ 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， ２８．２７， ２７．９２， ２５．７０， ２５．３０． 2) SPIB00201: L-serine-phenofibric acid ester Fenofibric acid (11.6 g, 36.3 mmol) cooled in an ice water bath, N-carbobenzyloxy-L-serine t-butyl ester (Boc-Ser- To a mixture of OtBu, 8.62 g, 33.0 mmol), EDC (7.59 g, 39.6 mmol) and DMAP (484 mg, 3.96 mmol) was added anhydrous dichloromethane (150 mL) dropwise. After completion of the addition, the ice water bath was removed, and the reaction mixture was stirred at room temperature for 20 hours under an argon atmosphere. After 20 hours, additional dichloromethane (200 mL) was added and the solution was washed with water (2 × 200 mL) and brine (200 mL). After drying over sodium sulfate and filtration, the solution was concentrated under reduced pressure. The remaining yellow oil (21.2 g) was purified by column chromatography using silica gel (400 g, 0.035 to 0.070 mm, 6 nm pore size) eluting with heptane / ethyl acetate (3: 1). The product-containing fractions were concentrated under reduced pressure and dried under high vacuum until the weight was constant, and the experiment showed that the protected L-serine-fenofibric acid ester SPIB0020101 (16.2 g, collected as a pale yellow oil). Rate 87%).

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, 28.27, 27.92, 25.70, 25.30.

^１Ｈ 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）．

^１３Ｃ 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分析：
１００％ ｐｕｒｉｔｙ； ｒ．ｔ． ＝ 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分析：
ｃａｌｃ． ： Ｃ ５４．３１， Ｈ ４．７９， Ｎ ３．１７ ； ｆｏｕｎｄ： Ｃ ５４．３７， Ｈ ４．７８， Ｎ ３．１２．

融点： １５１ ℃ （ｄｅｃ． ） A solution of protected L-serine-fenofibric acid ester SPIB0020101 (16.2 g, 28.8 mmol) in anhydrous dichloromethane (100 mL), cooled to 5 ° C. and stirred, under an argon atmosphere with an acetic acid solution of hydrogen chloride ( 400 mL, 1M, 400 mmol) 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 vacuum to remove acetic acid. Ethyl acetate (100 mL) was added to the remaining pale yellow oil (24.7 g). The solution was subjected to a second concentration and drying. 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 to room temperature. The precipitate was filtered off and dried under high vacuum at room temperature overnight and then at 43 ° C. for 1 hour. From the experiment, L-serine-fenofibric acid ester / hydrochloride SPIB00201 (7.66 g, yield 60%) was obtained 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; r. t. = 4.361 min. 55% TFA (0.1%), 45% ACN; 1 mL / min; 32.3 C, Luna C18, serial # 167717-13; 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.)

^１Ｈ 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）．

^１３Ｃ 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， ２７．９９， ２６．４４， ２４．７９， １６．９０． 3) SPIB00202: L-threonine-fenofibric acid ester Fenofibric acid (25.5 g, 79.9 mmol) cooled in an ice water bath, N-carbobenzyloxy-L-threonine t-butyl ester (Boc-Thr- OtBu, 20.0 g, 72.6 mmol, prepared by literature methods, to a mixture of EDC (16.7 g, 87.1 mmol) and DMAP (1.06 g, 8.71 mmol) in anhydrous dichloromethane (200 mL) After the addition was complete, the ice water bath was removed and the reaction mixture was stirred at room temperature under an argon atmosphere for 20 hours, after which additional EDC (1.39 g, 7.26 mmol) was added. In addition, the experimental solution was kept under stirring at room temperature under an argon atmosphere over the weekend, after 4 days additional dichloromethane (300 L) was added and the solution was washed with water (300 mL) and brine (300 mL) After drying over sodium sulfate and filtration, the solution was dried under reduced pressure, and the remaining yellow oil (53.5 g) was added. Purified by column chromatography using silica gel (500 g, 0.035 to 0.070 mm, 6 nm pore size) eluting with heptane / ethyl acetate (3: 1) .The product containing fractions were concentrated under reduced pressure, When dried under high vacuum until the weight was constant, the experiment gave protected L-threonine-fenofibric acid ester SPIB0020201 (34.1 g, 82% yield) as a white foam.

¹ H NMR (300 MHz, CDC 1 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, CDC 1 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, 27.99, 26.44, 24.79, 16.90 .

^１Ｈ 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）．

^１３Ｃ 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分析：
９８．５９％ ｐｕｒｉｔｙ； ｒ．ｔ． ＝ 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分析：
ｃａｌｃ．： Ｃ ５５．２７， Ｈ ５．０８， Ｎ ３．０７ ； ｆｏｕｎｄ： Ｃ ５４．９８， Ｈ ５．１３， Ｎ ３．０３．

融点： １６０．５ ℃ （ｄｅｃ． ） To 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. and stirred, under an argon atmosphere, an acetic acid solution of hydrogen chloride ( 600 mL, 1M, 600 mmol) was added dropwise. The reaction mixture was placed at 5 ° C. for 6 days. The mixture was concentrated under reduced pressure and dried under high vacuum to remove acetic acid. To the remaining white solid (45.8 g) was added ethyl acetate (500 mL). The mixture was heated to reflux for 10 minutes and then cooled to room temperature. The precipitate was filtered off and dried overnight at room temperature under high vacuum. From the experiment, L-threonine-fenofibric acid ester / hydrochloride SPIB00202 (26.3 g, yield 97%) was obtained 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, 79.99, 69.40, 55.21, 25.59, 24.22, 16.06.

HPLC analysis:
98.59% purity; r. t. = 4.687 min. 55% TFA (0.1%), 45% ACN; 1 mL / min; 32.3 C, Luna C18, serial # 167717-13; 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 ° C. (dec.)

^１Ｈ 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）．

^１３Ｃ 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， ７３．２６， ５８．２３， ５１．８８， ５１．５８， ３６．３３， ３５．３１， ３１．９２， ２８．２９， ２８．００， ２５．８９， ２４．９５． 4) SPIB00203: L-hydroxyproline-fenofibric acid ester Fenofibric acid (24.9 g, 78.1 mmol) cooled in an ice water bath, N-carbobenzyloxy-L-hydroxyproline t-butyl ester (Boc- Hyp-OtBu, 20.4 g, 71.0 mmol, prepared according to literature procedures, a mixture of EDC (16.3 g, 85.2 mmol) and DMAP (1.04 g, 8.52 mmol) in anhydrous dichloromethane ( After the addition was complete, the ice water bath was removed and the reaction mixture was stirred at room temperature for 20 hours under an argon atmosphere, after which additional EDC (1.63 g, 8.52 mmol) was added. The experimental solution was kept under stirring at room temperature under an argon atmosphere over the weekend. And then washed with sodium sulfate (200 mL), dried over sodium sulfate and filtered, then the solution was dried under reduced pressure, the remaining yellow oil (49.4 g) was added to silica gel (500 g, 0.035 to Purified by column chromatography eluting with heptane / ethyl acetate (2: 1) using 0.070 mm, 6 nm pore size) The product-containing fractions were concentrated under reduced pressure and kept under high vacuum until the weight was constant. As a colorless oil from the experiment, protected L-hydroxyproline-fenofibric acid ester SPIB0020301 (26.4 g, 63% yield) was obtained.

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 (0.62 H, t, J = 7.8 Hz), 3.67 (1.62 H, m), 3.46 (0.38 H, 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, CDC 1 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, 73.26, 58.23, 51.88, 51.58 , 36.33, 35.31, 31.92, 28.29, 28.00, 25.89, 24.95.

^１Ｈ 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）．

^１３Ｃ 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， ２４．４９．

ＨＰＬＣ分析：
１００％ ｐｕｒｉｔｙ ； ｒ．ｔ． ＝ ８．３６９ ｍｉｎ．； ６０％ ＤＩＵＦ ｗａｔｅｒ （０．１％ ＴＦＡ）／４０％ ａｃｅｔｏｎｉｔｒｉｌｅ； １ ｍＬ／ｍｉｎ； ３６．４ Ｃ； Ｌｕｎａ Ｃ１８， 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

融点： １８７．５ ℃ （ｄｅｃ．） To a stirred solution of protected L-hydroxyproline-fenofibric acid ester SPIB0020301 (26.0 g, 44.2 mmol) in anhydrous dichloromethane (100 mL) at 5 ° C. under an argon atmosphere with hydrogen chloride in acetic acid (100 mL). 450 mL, 1 M, 450 mmol) solution was added dropwise. The reaction mixture was stirred for 4 days at 5 ° C. After 4 days, the mixture was concentrated under reduced pressure and dried under high vacuum to remove acetic acid. To the remaining yellow oil (31.5 g), ethyl acetate (200 mL) was added. The mixture was sonicated and then concentrated under reduced pressure and dried under high vacuum. To the remaining white solid (23.2 g), ethyl acetate (300 mL) was added. The ethyl acetate mixture was refluxed for 10 minutes with heating and then cooled to room temperature. The precipitate was filtered off and dried overnight at room temperature under high vacuum. From the experiment, L-hydroxyproline-fenofibric acid ester / hydrochloride SPIB00203 (15.8 g, yield 76%) was obtained 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, 78.9, 73.79, 57.54, 50.23, 34.13, 25.69, 24.49.

HPLC analysis:
100% purity; r. t. = 8.369 min. 60% DIUF water (0.1% TFA) / 40% acetonitrile; 1 mL / min; 36.4 C; Luna C18, 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 ° C. (dec.)

The solubility of the ester was determined by excessively dissolving each drug in water at room temperature and then precipitating them for several hours. The resulting solution was centrifuged at 1500 rpm for 3 minutes, and the supernatant was analyzed. It has been found that these esters have a solubility in water of more than 50 mg / mL.

Experimental rats were examined for blood triglyceride levels at zero. The rats were then raised for 1 week on a high sugar-containing diet such as 30% sucrose water. Thereafter, at the end of the first week, the rats were examined for triglycerides and returned to normal diet. From day 7 to day 14, rats were administered test and control drugs. On day 14, the triglycerides in the rat blood were again examined.

In comparison of fenofibrate (control) and fenofibric acid L-serine ester (test drug), each of three rats was tested for equivalent doses of drug, control and 50, 100 and 200 mg / kg.

The results are shown below.

上記の結果より、高い水溶性のセリンエステルは、効果的に機能していると結論できる。 Summary-Dose Range Exploration Study-Hypolipoproteinemia Properties-Fenofibrate and its Formulation Test Material: L-Serine Ester Vehicle of Fenofibric Acid: 1% Tween 80 MilliQ Aqueous Solution

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

There are many screening tests that determine the usefulness of prodrugs created according to the disclosed methods, including both in vitro and in vivo screening methods.

In vitro methods include acid / base hydrolysis of prodrugs, hydrolysis in porcine pancreas, hydrolysis in rat intestinal fluid, hydrolysis in human gastric fluid, hydrolysis in human intestinal fluid and hydrolysis in human plasma Is mentioned. These assays are described in Simmons, DM, Chandran, VR and Portmann, GA, Danazol Amino Acid Prodrugs, In Vitro and In Situ Biopharmaceutical Evaluation, Drug Evelopment, Drug Development. The entire contents of which are incorporated by reference.

The prodrugs of fibric acid of the present invention are effective in the treatment of diseases or conditions where fibric acid derivatives are commonly used. The prodrugs disclosed herein can be converted in the body to release the active ingredient, thereby reducing or eliminating the biopharmaceutical and pharmacokinetic obstacles associated with fibric acid derivatives, thereby providing a therapeutic benefit for fibric acid. To strengthen. However, it should be noted that these prodrugs themselves have sufficient activity in mammals without releasing the active agent.

Thus, the prodrugs of the present invention enhance the therapeutic benefit by removing the biopharmaceutical and pharmacokinetic obstacles of existing drugs.

Furthermore, prodrugs are easily synthesized in high yields using readily available commercial reagents.

The prodrugs of fibric acid of the present invention are effective in the treatment of diseases or conditions where fibric acid derivatives are commonly used. The prodrugs disclosed herein can be converted in the body to release the active ingredient, thereby reducing or eliminating the biopharmaceutical and pharmacokinetic obstacles associated with fibric acid derivatives, thereby providing a therapeutic benefit for fibric acid. To strengthen. However, it should be noted that these prodrugs themselves have sufficient activity in mammals without releasing the active agent.

Thus, the fibric acid prodrug of the present invention enhances therapeutic benefits by removing the biopharmaceutical and pharmacokinetic obstacles of existing drugs. Furthermore, prodrugs are easily synthesized in high yields using readily available commercial reagents.

In the foregoing formulas and claims, AA shall be construed as defined below with the following content.

この定義のＡＡは、主鎖または側鎖上のいずれにもアミノ基を持たないアミノ酸残基を表す。

AA in this definition represents an amino acid residue having no amino group on either the main chain or the side chain.

この定義のＡＡは、側鎖上にヒドロキシ基を持たないアミノ酸残基を表す。

AA in this definition represents an amino acid residue having no hydroxy group on the side chain.

ＡＡは、主鎖または側鎖のいずれにもカルボキシ基を持たないアミノ酸基を表す。

AA represents an amino acid group having no carboxy group in either the main chain or the side chain.

であり、式中のＲｏは上記定義した側鎖アミノ酸である。 4) OAA—This is an ester bond between the hydroxy group of the drug and the carboxy group of an amino acid on the main chain or side chain. Therefore, when expressed in a formula, OAA is

Where Ro is a side chain amino acid as defined above.

Alternatively, it may represent an ester bond between the carboxy group of the drug and the hydroxy group on the side chain of an amino acid having a hydroxy group thereon, such as threonine, serine, hydroxyproline, tyrosine. The hydroxy group forms part of the ester bond represented above with O. Therefore, when expressed in the formula, AA represents an amino acid having a hydroxy group on the side chain. However, when expressed as OAA, AA does not have a hydroxy group because an oxygen atom is drawn in the formula.

Although the invention has been described in conjunction with the detailed description thereof, it is to be understood that the above description is intended to illustrate the invention and not to limit its scope. The scope of the invention is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Based on acetylcholine antagonism induced in albino mice, (±) ibuprofen L-serine ester (F1), L-threonine ester (±) ibuprofen (F2) and (±) ibuprofen L- It is a comparison figure of the effect of hydroxyproline ester (F3), (±) ibuprofen (that is, racemic mixture) and ibuprofen (S) (+). Based on acetylcholine antagonism induced in albino mice (±) ibuprofen L-serine ester (F1), (I) ibuprofen L-threonine ester (F2), (±) ibuprofen L -Comparison of potency with hydroxyproline ester (F3), ± ibuprofen and S (+) ibuprofen. It is the figure which represented the relationship of the dose response with respect to the mean coagulation time (MCT) of the L- serine ester of acetylsalicylic acid (formulation 1) in minutes. It is the figure which represented the relationship of the dose response with respect to the mean coagulation time (MCT) of the L-hydroxy proline ester of acetylsalicylic acid (formulation 2) in minutes. It is the figure which represented the relationship of the dose response with respect to the average coagulation time (MCT) of the L-threonine ester of acetylsalicylic acid (formulation 3) in minutes. It is the figure which represented the relationship of the dose response with respect to the average coagulation time (MCT) of a control (acetylsalicylic acid) in minutes. The relative efficacy of 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 (PC), FIG. 6 compares as a function of average clotting time expressed in minutes.

Claims (18)

A method for enhancing at least two therapeutic properties of an agent having a functional group selected from the group consisting of hydroxy, amino, or carboxy, wherein the improved therapeutic properties are:
(A) improved taste or aroma;
(B) desirable octanol / water partition coefficient;
(C) improved stability;
(D) increased blood-brain barrier permeability;
(E) elimination of the first-pass effect in the liver;
(F) reduced intrahepatic circulation;
(G) painless injection of parenteral formulations;
(H) improved bioavailability;
(I) improved absorption rate change;
(J) reduced side effects;
(K) dose proportionality;
(L) selective hydrolysis of the prodrug at the site of action;
(M) controlled release characteristics;
(N) targeted drug delivery;
(O) reduced toxicity;
(P) dose reduction;
(Q) alteration of metabolic pathways to increase the drug delivered to the site of action (r) increased solubility in aqueous solution; and (s) enhanced potency;
Wherein the method comprises (a) reacting the drug with an amino acid to form a covalent bond between the drug and the amino acid, wherein the drug is acetaminophen, adefovir, amlodipine , Amoxiline, Amphotericin B, Aspirin, Atorvastatin, Atobacon, Baclofen, Benazepril, Bexarotene, Candesarten, Capacitabine, Captopril, Carprofen, Carbidopa, Carboprost, Cefdinir, Cefditoren, Cefodiclozole, Cefpodoxime, Cefpodoxime , Diclofenac, didanosine, divalprox, diflunisal, 5-aminosalicylic acid, etodolac, efavirenz, enalapril, ep Lenone, eprosartan, estramustine, etodolac, ethosuximide, etotoin, etidocaine, etoposide, ezetimibe, fenofibrate, fenoprofen, fibric acid derivative, flavoxate, flurbiprofen, fluvastatin, focinopril, flovatriptan, flu Bestland, glimepiride, goserelin, gemfibrozil, ibuprofen, indomethacin, ketoprofen, ketorolac, lansoprazole, lopinavir, lovastatin, medroxyprogesterone, mefloquine, megestrol, metformin, methylphenidate, methylprednisolone, mexiletine, miglitol, moexril Naproxen, Naratriptan, Nelfinavir, Niacin, Nisolji , Norgestimate, octreotide, ofloxacin, olmesartan, omeprazole, pravastatin, perindopril, propofol, propoxyphene, ramipril, ritonavir, rosaprostol, salicylic acid, salmeterol, salsalate, salazine, sulindac, sertraline, simvastatin, sulphazine, Selected from the group consisting of sumitriptan, tolmetine, tazarotene, tenofovir, tolmetine, trandolapril, treprostinil, unoprostone, valproic acid, valsartan, zileuton, and zolmitriptan, or any pharmaceutically acceptable salt of said drug The amino acid is threonine or L-threonine.   The drug is cyclosporine, lopinavir, ritonavir, cefdinir, zileuton, nelfinavir, flavoxate, candesarten (Candesarten), propofol, nisoldipine, amlodipine, ofloxacin, fosinopril, enalapril, ramipril, benazepril, prindomol pril, prindopril p , Amoxiline, cefuroxime, ceftazidime, cefpodoxime, atovacon, niacin, bexarotene, propoxyphene, salsalate, acetaminophen, ibuprofen, lovastatin, simvastatin, atorvastatin, pravastatin, fluvastatin, nadolol, valsartan, methylphenidate, sulfa drug, 5- amino Lithic acid, sulfasalazine, methylprednisolone, medroxyprogesterone, estramustine, miglitol, mefloquine, capacitabine, danazol, eprosartan, valproic acid, omeprazole, lansoprazole, megestrol, metformin, tazarotene, sumitriptan , Zolmitriptan, aspirin, olmesartan, clopidogrel, amphotericin, tenofovir, unoprostone, fulvestrant, cefditorene, efavirenz, eplerenone, treprostinil, adefovir, salicylic acid, diflunisal, fenoprofen, carprofen, flurprofen Etodolac, sulindac, indomethacin, Rumechin, ketorolac, Sarajin or gemfibrozil, or method according to claim 1, characterized in that any pharmaceutically acceptable salt of said agent,.   Reaction of the functional group with the amino acid or a pharmaceutically acceptable salt thereof by a reaction between a drug having a functional group selected from the group consisting of hydroxy, amino, and carboxy and an L-α-amino acid Wherein the drug is a cyclosporine, lopinavir, ritonavir, cefdinir, zileuton, nelfinavir, flavoxate, candesarten, propofol, nisoldipine, amlodipine, ofloxacin, focinopril, enalapril, ramipril, peripril , Trandolapril, cromolyn, amoxiline, cefuroxime, ceftazidime, cefpodoxime, atobacon, niacin, bexarotene, propoxyphene, salsalate, acetaminophen Ibuprofen, lovastatin, simvastatin, atorvastatin, pravastatin, fluvastatin, nadolol, valsartan, methylphenidate, sulfa drugs, 5-aminosalicylic acid, sulfasalazine, methylprednisolone, medroxyprogesterone, estramustine, miglitol, mefloquine, capacitabine, danazol, ep Losartan, valproic acid, omeprazole, lansoprazole, megestrol, metformin, tazarotene, sumitriptan, naratriptan, zolmitriptan, aspirin, olmesartan, clopidogrel, amphotericin, tenofovir, unoprostone, fulvestrant, cefditoren , Adefovir, salicylic acid, diflu Nisal, fenoprofen, carprofen, flurbiprofen, ketoprofen, naproxen, etodolac, sulindac, indomethacin, tolmetin, ketorolac, salazine, or genfibrozil, or any pharmaceutically acceptable salt of the drug, A product characterized in that the amino acid is threonine or L-threonine.   The drug is cyclosporine, lopinavir, cefdinir, zileuton, nelfinavir, flavoxate, candesarten, propofol, nisoldipine, amlodipine, ofloxacin, enalapril, ramipril, focinopril, benazepril, perindopril, moexipril, osmolin p Cefuroxime, ceftazidime, cefpodoxime, atovacon, niacin, bexarotene, propoxyphene, salsalate, acetaminophen, ibuprofen, lovastatin, simvastatin, atorvastatin, pravastatin, fluvastatin, nadolol, valsartan, methylphenidate, sulfa drug, sulfasalazine, methylprednisolone Droxyprogesterone, estramustine, miglitol, mefloquine, capacitabine, danazol, eprosartan, valproic acid, gabapentin, omeprazole, lansoprazole, megestrol, metformin, tazarotene, sumitriptan, naratriptan, zolmitriptan, olmesartan, aspirin, Clopidogrel, amphotericin, tenofovir, unoprostone, fulvestrant, cefditorene, efavirenz, eplerenone, salicylic acid, 5-aminosalicylic acid, diflunisal, fenoprofen, carprofen, flurbiprofen, ketoprofen, naproxen, etodolactin, indolmethine, indolmethine tin Ketorolac, genfibrozil, or salazine, or the above A product according to claim 3, characterized in that any pharmaceutically acceptable salt of the agent. formula:

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Sulfo drug azo derivative R ₈ N═N—AA ₅ —COOH;
Sulfur drug derivative _{R 8 NH CO-AA 5 NH} 2; or

;
Or a pharmaceutically acceptable salt thereof;
Product of;
AA is an amino acid having no hydroxyl group on the carboxy group, and is bonded via the acyl group;
R is AA ₃ or OAA ₆ ;
R ₁ is an amino acid without an amino group and a carboxy group;
CYCLO represents a residue at positions 2 to 11 of the cyclosporine molecule;
x-y is located in the CH = CH or _{CH 2} = _CH 2;
R ² is OAA or OGlyAA;
AA ₁ is an amino acid having no hydroxy group on the carboxy group, and is bonded via the acyl group;
AA ₆ is an amino acid residue having fewer side chain hydroxy groups than the hydroxyl group, wherein AA ₆ is linked by an ester linked through the side chain hydroxy group;
R ₂ is:

;

Or

;
Is;
R ₅ is AA ₂ ;
AA ₂ is an amino acid residue having fewer hydroxy groups than on the carboxy group, and is bonded via the acyl group;
AA ₃ is an amino acid having fewer hydrogen atoms than on the amino group, and is bonded via the amino group;
R ₆ is AA ₁ ;
R ₇ is O-AA ₁ ;
AA ₅ is an amino acid without an amino group and a carboxy group;
R ₈ is a sulfanilamide component of a general class of sulfa drugs;
R ₉ is AA ₃ ;
AA, AA ₁ , AA ₂ , AA ₃ , AA ₅ , and AA ₆ are threonine or L-threonine;
A product characterized by that. The compound has the formula:

The product according to claim 5, wherein   A pharmaceutical composition comprising a therapeutically effective amount of the product of any one of claims 5 or 6 and a pharmaceutically acceptable salt thereof.   A method for enhancing the bioavailability of a drug having a functional group selected from the group consisting of hydroxy, amino, or carboxy, wherein the drug is cyclosporine, lopinavir, cefdinir, zileuton, nelfinavir, flavoxate, candesarten, propofol , Nisoldipine, amlodipine, ofloxacin, fosinopril, enalapril, ramipril, benazepril, moexipril, trandolapril, cromolyn, amoxiline, cefuroxime, ceftazidime, cefpodoxime, atovacon, ganciclovir, penciclovir, famciclovir, acyclobecin Salsalate, acetaminophen, ibuprofen, lovastatin, Nvastatin, atorvastatin, pravastatin, fluvastatin, nadolol, valsartan, methylphenidate, methylprednisone, sulfa drugs, sulfasalazine, medroxyprogesterone, estramustine, 5-aminosalicylic acid, miglitol, mefloquine, danazol, eprosartan, dival Prox, fenofibrate, gabapentin, omeprazole, lansoprazole, megestrol, metformin, tazarotene, sumatriptan, naratriptan, zolmitriptan, aspirin, olmesartan, sirolimus, tacrolimus, pimecrolimus, clopidogrel, amphotericin, tenofovir, tenofovir Runt, Cefditoren, Efavirenz, Eplerenone, Trepros Selected from the group consisting of nil or adefovir, wherein the method comprises reacting the drug and an amino acid to form a covalent bond between the drug and the amino acid, wherein the amino acid is threonine or L- A method characterized by being threonine.   A method for enhancing the solubility in aqueous solution of a drug having a functional group selected from the group consisting of hydroxy, amino, or carboxy, wherein the drug is lopinavir, cefdinir, amlodipine, nisoldipine, zileuton, nelfinavir, flavoxate, candesartetane, cromolyn ( Sodium), atobacon, niacin, bexarotene, propoxyphene, salsalate, acetaminophen, ibuprofen, valsartan, methylphenidate, methylprednisolone, medroxyprogesterone, cephalosporins, estramustine, miglitol, mefloquine, danazol , Eprosartan, divalplex, fenofibrate, gabapentin, megestrol, metformin, tazarotene, aspirin, Selected from the group consisting of lumesartan, clopidogrel, amphotericin, tenofovir, unoprostone, fulvestrant, cefditoren, efavirenz, eplerenone, treprostinil, adefovir and salazine, and the method comprises the drug and the amino acid, Reacting to form a covalent bond therebetween, wherein the amino acid is threonine or L-threonine.   A method for enhancing safety properties having a longer anesthetic action in an aqueous solution of propofol, wherein the hydroxy functionality of the propofol molecule and an amino acid are reacted to form a covalent bond between the propofol and the amino acid Comprising the step, wherein the amino acid is threonine or L-threonine.   A method for increasing the solubility of a quinolone antibiotic having a carboxy group in an aqueous solution, wherein the carboxylic acid functional group of the quinolone antibiotic and an amino acid are covalently bonded between the amino acid and the antibiotic. Reacting to form, wherein the amino acid is threonine or L-threonine.   A method for increasing the solubility of an ACE inhibitor having a carboxy group or an acylating group in an aqueous solution, wherein a carboxylic acid functional group of the ACE inhibitor molecule and an amino acid are covalently bonded between the ACE inhibitor and the amino acid. And the amino acid is threonine or L-threonine.   A method for increasing the solubility of a proton pump inhibitor in an aqueous solution, comprising the step of reacting an amine functional group of the proton pump inhibitor with a carboxylic acid moiety of an amino acid so as to form an amide bond. A method characterized by being threonine or L-threonine.   A method for increasing the solubility of a selective agonist of 5-HT receptor in an aqueous solution, wherein an amine function of the selective agonist molecule of 5-HT receptor and a carboxylic acid moiety of an amino acid form an amide. The amino acid is threonine or L-threonine.   A method for increasing the solubility of an immunosuppressive agent in an aqueous solution, comprising the step of reacting the hydroxy functional group of the immunosuppressive agent with an amino acid so as to form an ester, wherein the amino acid is threonine or L-threonine. A method characterized in that   A method for enhancing the solubility of a sulfa drug in an aqueous solution, comprising the step of reacting an amine of the sulfa drug with an amino acid so as to form an amide, wherein the amino acid is threonine or L-threonine. Feature method.   A method for providing a transparent water-soluble intravenous injection formulation containing a sulfa drug, wherein the amine of the sulfa drug and an amino acid are reacted so as to form a covalent bond between the sulfa drug and the amino acid. And the amino acid is threonine or L-threonine.   A method for increasing the solubility of an OH functional group-containing nucleoside analog in an aqueous solution, wherein the hydroxyl functional group of the nucleoside analog and an amino acid form an ester between the nucleoside analog and the amino acid. And the amino acid is threonine or L-threonine.

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