KR100472303B1 - Corticosteroid-21-sulfate sodium derivatives as colon-specific prodrugs - Google Patents

Abstract

The present invention relates to a corticosteroid 21-sulfate sodium derivative of Formula 1 useful as a colon-specific prodrug of a corticosteroid, a method for preparing the same, and an inflammatory bowel disease containing the same (Inflammatory) Bowel Diseases (IBD) relates to a pharmaceutical composition for the prevention or treatment of:

Where , R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined in the specification.

Description

Corticosteroid-21-sulfate sodium derivatives as colon-specific prodrugs

The present invention relates to a corticosteroid 21-sulfate sodium derivative of Formula 1 useful as a colon-specific prodrug of a corticosteroid, a method for preparing the same, and an inflammatory bowel disease containing the same (Inflammatory) Bowel Diseases (IBD) relates to a pharmaceutical composition for the prevention or treatment of:

[Formula 1]

Where

Represents a single or double bond,

R ¹ represents hydrogen or CH ₃ ,

R ² represents hydrogen or F,

R ³ represents ═O or OH,

R ⁴ represents hydrogen, OH or CH ₃ ,

R ⁵ represents OH, or

R ⁴ and R ⁵ together Indicates.

Inflammatory bowel disease, which is classified as ulcerative colitis and Crohn's disease, is a refractory disease and has a high recurrence rate after treatment. Up to now, it has been recognized mainly in the West, but since late 1980, it has been increasing in Asia including Korea. Until now, their exact etiology is not known, and when symptomatic treatment is very severe, salicylate derivatives are used as a maintenance therapy to relieve symptoms and prevent relapse by administering large amounts of corticosteroids. Corticosteroids have strong anti-inflammatory properties and are effective in attack therapy of ulcerative colitis. However, since it is absorbed well in the upper digestive tract, it does not reach the end of the small intestine or the colon, so that only a part of the dosage reaches the affected part, which requires a large amount of administration. In addition, long-term use may cause serious side effects such as osteoporosis, hypertension, decreased immunity, diabetes, etc., due to the systemic absorption of the upper digestive tract. Therefore, in order to treat a disease occurring in the colon, a drug-targeted drug delivery method that selectively delivers a drug to a focal colon is an essential task to increase the treatment effect and reduce systemic side effects.

Colon targeting drug delivery can be roughly divided into those using a pharmaceutical formulation and those using a prodrug. In the former case, the gastric emptying time is not constant, and the pH of the large intestine and the small intestine does not differ so much that selective delivery is difficult. Therefore, recently, a method for preparing a drug in the form of a prodrug to safely reach the colon area has been extensively studied.

The method of using a prodrug is basically a method of binding a drug to a polymer or a hard-absorbing water-soluble compound as a carrier so that the prodrug is not absorbed from the upper digestive tract. In this case, the binding between the carrier and the drug must be chemically or biologically stable while passing through the upper digestive tract, so that it can be selectively delivered to the large intestine, and must be delivered after being delivered to the large intestine so that the effective drug can be released. Therefore, as a result of recognizing that enzymes secreted by microorganisms living in human colon can activate prodrugs, research on colonic target drug delivery controlled by microorganisms is being actively conducted.

To date, little is known about the colon-targeting corticosteroids used in the treatment of inflammatory bowel disease, and dextran or the hydrophilic small molecule glucuronic acid, which has recently been characterized as a colon-targeting carrier. Or prodrugs that combine drugs with glucose or the like has been reported. Colonic target prodrugs of corticosteroids should release the active drug, corticosteroid, from the follicle's colon and directly act on the affected area, and in the upper digestive tract to minimize side effects caused by systemic absorption of the drug. Should have a low absorption. Dexamethasone-dextran is a compound that has been reported as a colon-targeting prodrug of corticosteroids, which has a disadvantage in that it is not activated in the large intestine when the degree of substitution of the drug that binds to dextran is large. The synthesis process is very complex, and it is currently unknown whether they have reached clinical stage of use. In conclusion, the development of colon-targeted corticosteroid prodrugs that can effectively treat inflammatory bowel disease that requires long-term administration without side effects is urgently needed.

The present inventors attempted to develop a sodium target of sulfate ester on the hydroxyl position 21 of the corticosteroid to develop a colon target prodrug of the corticosteroid. Corticosteroid 21-sulfate sodium was expected to inhibit absorption in the digestive tract due to the introduction of a polar sulphate sodium group to increase the polarity of the compound, and sulfate ester groups are stable to enzymes in the upper digestive tract. Absorption was suppressed and stable throughout the upper digestive tract, so most of the dose was expected to be delivered to the affected colon. In addition, the presence of sulfatase, an enzyme secreted from the intestinal microorganisms, breaks down the sulfate ester group, so when corticosteroid 21-sulfate sodium reaches the colon, it is hydrolyzed to produce corticosteroids, which act directly on the inflammation of the colon. It was expected to have an excellent therapeutic effect.

The present inventors synthesized sulfonic acid ester sodium salts using various corticosteroids as starting materials, and tested their effects as colon target prodrugs. As a result, it was confirmed that the corticosteroid 21-sulfate sodium salt of Formula 1 can effectively achieve the above object, and completed the present invention.

First, the present invention relates to corticosteroid 21-sulfate sodium derivatives of formula (I):

[Formula 1]

Where

Represents a single or double bond,

R ¹ represents hydrogen or CH ₃ ,

R ² represents hydrogen or F,

R ³ represents ═O or OH,

R ⁴ represents hydrogen, OH or CH ₃ ,

R ⁵ represents OH, or

R ⁴ and R ⁵ together Indicates.

Secondly, the present invention comprises the step of reacting the corticosteroid of formula (2) with sulfur trioxide triethylamine of formula (3) to obtain a corticosteroid 21-sulfate triethylammonium salt of formula (4), and then reacting it with a saturated saline solution. A method for preparing a corticosteroid 21-sulfate sodium derivative of formula (I):

[Formula 2]

[Formula 3]

_{SO 3 · N (C 2 H} 5) 3

[Formula 4]

Where , R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined above.

Third, the present invention relates to a pharmaceutical composition for preventing or treating inflammatory bowel disease, which contains a corticosteroid 21-sulfate sodium derivative of formula (1) as an active ingredient.

Hereinafter, the present invention will be described in detail.

In the present invention, more preferred compounds are Corticosteroid 21-sulfate of formula (I) which represents this double bond, R ¹ is H, R ² is H or F, R ³ is OH, R ⁴ is H or α-CH ₃ , and R ⁵ is OH Sodium derivative (ie dexamethasone 21-sulfate sodium or prednisolone 21-sulfate sodium).

Embodiments of the compounds according to the invention are as follows:

Dexamethasone 21-sulfate sodium: Dexa-S: C ¹ = C 2, R ¹ ; H, R ² ; F, R ³ ; OH, R ⁴ ; α-CH ₃ , R ⁵ ; OH);

Prednisolone 21-sulfate sodium (PD-S): C ¹ = C ² , R ¹ , R ² and R ⁴ ; H, R ³ and R ⁵ ; OH;

Methylprednisolone-21-sulfate sodium (MP-S): C ¹ = C 2, R ¹ ; CH ₃ , R ³ ; OH, R ² and R ⁴ ; H, R ⁵ ; OH;

Hydrocortisone-21-sulfate sodium (HC-S): C1-C2, R ¹ , R ² and R ⁴ ; H, R ³ and R ⁵ ; OH;

Cortisone 21-sulfate sodium (CR-S): C1-C2, R ¹ , R ² and R ⁴ ; H, R ³ ; = 0, R ⁵ ; OH;

Betamethasone-21-sulfate sodium: Beta-S: C1 = C2, R ¹ ; H, R ² ; F, R ³ ; OH, R ⁴ ; β-CH ₃ , R ⁵ ; OH;

Triamcinolone 21-sulfate sodium (TA-S): C ¹ = C 2, R ¹ ; H, R ² ; F, R ³ ; OH, R ⁴ ; β-OH, R ⁵ ; OH;

Fludrocortisone-21-sulfate sodium (FC-S): C1-C2, R ¹ and R ⁴ ; H, R ² ; F, R ³ and R ⁵ ; OH,

Budesonide-21-sulfate sodium (BD-S): C ¹ = C ² , R ¹ and R ² ; H, R ³ ; OH, R ⁴ and R ⁵ ; In combination .

In the present invention, the physicochemical properties, that is, the properties of solubility, chemical stability in the upper digestive tract, biochemical stability, refractory absorption, etc., and most of the dosage is delivered to the colon and at the same time reach the affected colon, the active drug In consideration of the conversion rate and the like, dexamethasone 21-sulfate sodium (Dexa-S) is most preferred.

The corticosteroid 21-sulfate sodium derivative of formula 1 according to the present invention is obtained by reacting the corticosteroid of formula 2 with sulfur trioxide triethylamine of formula 3 to obtain a corticosteroid 21-sulfate triethylammonium salt of formula 4, It can be obtained by reaction with a saturated saline solution:

[Formula 2]

[Formula 3]

_{SO 3 · N (C 2 H} 5) 3

[Formula 4]

Where

, R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined above.

The preparation method can be represented by the following scheme 1.

As shown in Scheme 1, in the process of the present invention, the corticosteroid of Formula 2 is reacted with sulfur trioxide triethylamine of Formula 3 in a first step to produce an intermediate of Formula 4. Sulfur trioxide pyridine may be used in place of sulfur trioxide triethylamine in the first stage reaction. However, sulfur trioxide triethylamine may be used in consideration of the economical efficiency, yield, and side reaction of the reaction. Particular preference is given to using.

In addition, the reaction may preferably be carried out in an organic solvent that does not adversely affect the reaction. Examples of organic solvents that can be used for this purpose include benzene, pyridine, tetrahydrofuran, dichloromethane, toluene, and the like, and preferably, the reaction is carried out in benzene or a mixed solvent of benzene and pyridine. The reaction is also preferably carried out under anhydrous conditions.

The reaction temperature and time are not particularly limited, and in general, the reaction may be performed for several minutes to several hours at room temperature to warm. Preferably the reaction is carried out at 50 to 60 ° C. for several minutes to several hours.

The compound of formula 4 produced by the first stage reaction is then converted to the sodium salt in the second stage to produce the compound of formula 1. In order to convert the compound of Formula 4 to sodium salt, an aqueous solution of the compound of Formula 4 is generally added to 10% saline to form a precipitate. In this case, it is preferable to use saturated saline to increase the yield.

After completion of the reaction according to the invention as described above, the product can be separated and purified by conventional workup methods such as chromatography, recrystallization and the like.

The corticosteroid 21-sulfate sodium derivative of formula 1 according to the invention is useful as a colon target corticosteroid prodrug. For example, dexamethasone 21-sulfate sodium is rarely absorbed in the upper digestive tract and is delivered to the large intestine and converted to the active drug dexamethasone by an enzyme secreted from the intestinal microorganisms, more than 90%. Generally, prodrugs are decomposed into active drugs and carriers after reaching the desired site of action. Therefore, the compounds used as carriers need to be safe in the body. However, dexamethasone 21-sulfate is suitable for the toxicological safety of carrier compounds. Since it is identical in structure to the normal metabolite of dexamethasone in the liver, it is not a toxicological problem at all. In the present invention, dexamethasone 21-sulfate sodium, in combination with its in vitro and in vivo experimental results, significantly lowers the degree of absorption in the upper digestive tract compared to the existing dexamethasone to reduce side effects and the amount of prodrugs that reach the large intestine It is an excellent colon target prodrug of dexamethasone that can increase the local concentration of dexamethasone by increasing the therapeutic effect.

Accordingly, the present invention also provides a pharmaceutical composition for the prophylaxis or treatment of inflammatory bowel disease such as ulcerative colitis and Crohn's disease, which contains a corticosteroid 21-sulfate sodium derivative of Formula 1, including dexamethasone 21-sulfate sodium derivative as an active ingredient. It is about.

When administering a compound of the present invention for clinical purposes, the total daily dose to be administered in the host in single or discrete doses is preferably in the range of 0.02 mg to 0.1 mg / kg body weight as a corticosteroid. However, the specific dose level for a particular patient may vary depending on the specific compound to be used, the weight of the individual patient, sex, health condition, diet, time of administration, method of administration, rate of excretion, drug mixing and the severity of the disease.

Compositions containing a compound of the present invention as an active ingredient may be combined with a conventional carrier in the pharmaceutical field for clinical use, such as tablets, capsules, troches, solutions, suspensions, etc. It can be formulated into various types of oral preparations. Carriers that can be used in the compositions of the present invention are conventional in the pharmaceutical field, for example in the case of oral preparations, binders, lubricants, disintegrants, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavorings Etc. The pharmaceutical preparations thus prepared may be formulated and administered in various forms for oral administration.

The present invention is explained in more detail by the following examples and experimental examples, but the scope of the present invention is not limited in any way by these.

Example 1 Synthesis of Dexamethasone 21-Sulfate Triethylammonium

3.92 g (10 mmol) of dexamethasone was added to anhydrous pyridine solution, and a small amount of sulfur trioxide triethylamine (5.58 g, 25 mmol) was added thereto and reacted at 56 to 60 ° C. for 2 hours with mechanical stirring. Thereafter, the reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain the title compound.

¹ H-NMR (D ₂ O): 1.00 (s, 3H, C-18), 1.21 (t, 3H), 1.55 (s, 3H, C-19), 3.13 (q, 2H), 4.90 to 5.21 ( AB q, 2H, C-21), 6.21 (s, 1H, C-4), 6.37 (d, 1H, C-1), 7.35 (d, 1H, C-2)

Example 2: Synthesis of Dexamethasone 21-Sulfate Sodium

The dexamethasone 21-sulfate triethylammonium obtained in Example 1 was dissolved in a minimum amount of distilled water and stirred while adding to a 10% sodium chloride solution. The resulting precipitate was separated by suction filtration, washed with cold ethanal and dried to give 4.2 g (84% yield) of the title compound.

IR (cm ^-1 , Newsol): 1719 (C = O), 1662, 1611 (C = C), 1259 (S = O), 1034 (S = O)

¹ H-NMR (D ₂ O): 1.01 (s, 3H, C-18), 1.55 (s, 3H, C-19), 4.90 to 5.05 (AB q, 2H, C-21), 6.23 (s, 1H, C-4), 6.43 (d, 1H, C-1), 7.54 (d, 1H, C-2)

Elemental Analysis (C ₂₂ H ₂₈ FSO ₈ Na): Calculated (C; 53.44, H; 5.67, S; 6.48), Found (C; 53.58, H; 5.10, S; 6.65)

Example 3: Synthesis of Prednisolone 21-Sulfate Triethylammonium Salt

3.6 g (10 mmol) of prednisolone was added to 64 mL of anhydrous benzene and pyridine mixed solvent (1/1), and 3.8 g (17 mmol) of sulfur trioxide triethylamine was added little by little with stirring to react for 20 minutes at 56 to 60 ° C. The solvent was then concentrated by evaporation under reduced pressure to afford the title compound.

¹ H-NMR (D ₂ O): 1.00 (s, 3H, C-18), 1.21 (t, 3H), 1.55 (s, 3H, C-19), 3.13 (q, 2H), 4.90, 5.21 ( AB q, 2H, C-21), 6.23 (s, 1H, C-4), 6.43 (d, 1H, C-1), 7.54 (d, 1H, C-2).

Example 4 Synthesis of Prednisolone 21-Sulfate Sodium

The prednisolone 21-sulfate triethylammonium salt obtained in Example 3 was stirred while added to a 10% sodium chloride solution. The precipitate thus formed was filtered and the filtrate was purified on an open silica gel column using a chloroform / methanol (7/3) solution as an eluting solvent to give 4.9 g (yield 71%) of the title compound.

IR (cm ^-1 , Newsol): 1675 (C = O), 1640 (C = C), 1257 (S = O), 1035 (S = O)

¹ H-NMR (D ₂ O): 0.78 (s, 3H, C-18), 1.32 (s, 3H, C-19), 4.80 to 4.98 (AB q, 2H, C-21), 5.96 (s, 1H, C-4), 6.22 (d, 1H, C-1), 7.42 (d, 1H, C-2)

Elemental Analysis (C ₂₁ H ₂₇ SO ₈ Na): Calculated (C; 54.55, H; 5.84, S; 6.93), Found (C; 54.27, H; 6.13, S; 6.64)

Example 5 Synthesis of Methylprednisolone 21-Sulfate Sodium

0.37 g (1.0 mmol) of methylprednisolone was added to 64 mL of anhydrous benzene and pyridine mixed solvent (1/1), and 0.38 g (1.7 mmol) of sulfur trioxide triethylamine was added little by little with stirring to react at 56 to 60 ° C for 10 minutes. . The solvent was then concentrated by evaporation under reduced pressure to afford methylprednisolone 21-sulfate triethylammonium salt. The obtained methylprednisolone 21-sulfate triethylammonium salt was added to the 10% sodium chloride solution while stirring. The resulting precipitate was filtered and the filtrate was purified by silica gel open column using chloroform / methanol (7/3) solution as eluent to yield 0.32 g (yield 67%) of the title compound.

IR (cm ^-1 , Newsol): 1670 (C = O), 1634 (C = C), 1255 (S = O), 1038 (S = O), ¹ H-NMR (D ₂ O): 0.76 (s , 3H, C-18), 1.37 (s, 3H, C-19), 4.85-4.92 (AB q, 2H, C-21), 5.84 (s, 1H, C-4), 6.27 (d, 1H, C-1), 7.32 (d, 1H, C-2)

Elemental Analysis (C ₂₂ H ₂₉ SO ₈ Na): Calculated (C; 55.46, H; 6.09, S; 6.72), Found (C; 55.27, H; 6.13, S; 6.34)

Example 6 Synthesis of Hydrocortisone 21-Sulfate Sodium

0.36 g (1.0 mmol) of hydrocortisone was added to 64 mL of anhydrous benzene and pyridine mixed solvent (1/1), and 0.38 g (1.7 mmol) of sulfur trioxide triethylamine was added little by little with stirring to react at 56 to 60 DEG C for 10 minutes. . Then, hydrocortisone 21-sulfate triethylammonium salt obtained by evaporating the solvent under reduced pressure was obtained. The obtained hydrocortisone 21-sulfate triethylammonium salt was added to the 10% sodium chloride solution while stirring. The resulting precipitate was filtered and the filtrate was purified on an open silica gel column using a chloroform / methanol (7/3) solution as an eluting solvent to give 0.32 g (yield 69%) of the title compound.

IR (cm ^-1 , Newsol): 1681 (C = O), 1250 (S = O), 1032 (S = O)

¹ H-NMR (D ₂ O): 0.80 (s, 3H, C-18), 1.40 (s, 3H, C-19), 4.54-4.60 (AB q, 2H, C-21), 5.59 (s, 1H, C-4)

Elemental Analysis (C ₂₁ H ₂₉ SO ₈ Na): Calculated (C; 54.31, H; 6.25, S; 6.90), Found (C; 54.07, H; 6.13, S; 6.54)

Example 7: Synthesis of Fludrocortisone 21-Sulfate Sodium

0.38 g (1.0 mmol) of fludrocortisone was added to 20 ml of anhydrous benzene and pyridine mixed solvent (1/1), and 0.38 g (1.7 mmol) of sulfur trioxide triethylamine was added little by little with stirring to react for 10 minutes at 56 to 60 ° C. I was. The solvent was then concentrated by evaporation under reduced pressure to give the fludrocortisone 21-sulfate triethylammonium salt. The obtained hydrocortisone 21-sulfate triethylammonium salt was added to the 10% sodium chloride solution while stirring. The resulting precipitate was filtered and the filtrate was purified by silica gel open column using chloroform / methanol (7/3) solution as eluent to yield 0.34 g (yield 70%) of the title compound.

IR (cm ^-1 , Newsol): 1721, 1674 (C = O), 1260 (S = O), 1035 (S = O)

¹ H-NMR (D ₂ O): 0.78 (s, 3H, C-18), 1.49 (s, 3H, C-19), 4.98-5.10 (AB q, 2H, C-21), 5.74 (s, 1H, C-4)

Elemental Analysis (C ₂₁ H ₂₈ FSO ₈ Na): Calculated (C; 52.39, H; 5.82, S; 6.65), Found (C; 52.07, H; 6.03, S; 6.31)

Example 8 Synthesis of Cortisone 21-Sulfate Sodium

0.36 g (1.0 mmol) of cortisone was added to 20 ml of anhydrous benzene and pyridine mixed solvent (1/1), and 0.38 g (1.7 mmol) of sulfur trioxide triethylamine was added little by little with stirring to react at 56 to 60 ° C for 10 minutes. The solvent was then concentrated by evaporation under reduced pressure to afford cortisone 21-sulfate triethylammonium salt. The resulting cortisone 21-sulfate triethylammonium salt was stirred with 10% sodium chloride solution. The resulting precipitate was filtered and the filtrate was purified by silica gel open column using chloroform / methanol (7/3) solution as eluent to yield 0.34 g (yield 73%) of the title compound.

IR (cm ^-1 , Newsol): 1725, 1674 (C = O), 1260 (S = O), 1035 (S = O)

¹ H-NMR (D ₂ O): 0.48 (s, 3H, C-18), 1.31 (s, 3H, C-19), 4.89-4.95 (AB q, 2H, C-21), 5.64 (d, 1H, C-4)

Elemental Analysis (C ₂₁ H ₂₈ SO ₈ Na): Calculated (C; 54.43, H; 6.05, S; 6.91), Found (C; 54.25, H; 6.23, S; 6.65)

Example 9 Synthesis of Triamcinolone 21-Sulfate Sodium

0.39 g (1.0 mmol) of triamcinolone was added to 24 ml of anhydrous benzene and pyridine mixed solvent (1/1), and 0.38 g (1.7 mmol) of sulfur trioxide triethylamine was added little by little with stirring to react for 10 minutes at 56 to 60 ° C. Thereafter, the solvent was evaporated under reduced pressure to give triamcinolone 21-sulfate triethylammonium salt. The obtained triamcinolone 21-sulfate triethylammonium salt was added to the 10% sodium chloride solution while stirring. The precipitate thus formed was filtered and the filtrate was purified by silica gel open column using a chloroform / methanol (7/3) solution as an eluting solvent to give 0.32 g (yield 62%) of the title compound.

IR (cm ^-1 , Newsol): 1719 (C = O), 1662, 1611 (C = C), 1257 (S = O), 1036 (S = O)

¹ H-NMR (D ₂ O): 0.84 (s, 3H, C-18), 1.49 (s, 3H, C-19), 4.89 to 5.02 (AB q, 2H, C-21), 6.02 (s, 1H, C-4), 6.23 (d, 1H, C-1), 7.27 (d, 1H, C-2)

Elemental Analysis (C ₂₁ H ₂₆ FSO ₉ Na): Calculated (C; 50.91, H; 5.25, S; 6.46), Found (C; 51.08, H; 5.10, S; 6.60)

Example 10 Synthesis of Betamethasone 21-Sulfate Sodium

0.39 g (1.0 mmol) of betamethasone was added to 24 ml of anhydrous benzene and pyridine mixed solvent (1/1), and 0.38 g (1.7 mmol) of sulfur trioxide triethylamine was added little by little with stirring to react at 56 to 60 ° C for 10 minutes. Thereafter, the betamethasone 21-sulfate triethylammonium salt obtained by evaporating the solvent under reduced pressure was added to a 10% sodium chloride solution, followed by stirring. At this time, the resulting precipitate was filtered and the filtrate was purified on an open silica gel column using a chloroform / methanol (7/3) solution as an eluting solvent to obtain 0.4 g (yield 81%) of the title compound.

IR (cm ^-1 , Newsol): 1719 (C = O), 1662, 1611 (C = C), 1257 (S = O), 1036 (S = O)

¹ H-NMR (D ₂ O): 0.84 (s, 3H, C-18), 1.49 (s, 3H, C-19), 4.89 to 5.02 (AB q, 2H, C-21), 6.02 (s, 1H, C-4), 6.23 (d, 1H, C-1), 7.27 (d, 1H, C-2)

Elemental Analysis (C ₂₂ H ₂₈ FSO ₈ Na): Calculated (C; 53.44, H; 5.67, S; 6.48), Found (C; 53.58, H; 5.10, S; 6.65)

Example 11: Synthesis of Budesonide 21-Sulfate Sodium

0.43 g (1.0 mmol) of budesonide was added to 25 ml of anhydrous benzene and pyridine mixed solvent (1/1), and 0.38 g (1.7 mmol) of sulfur trioxide triethylamine was added little by little with stirring to react for 10 minutes at 56 to 60 ° C. I was. The solvent was evaporated under reduced pressure to give budesonide 21-sulfate triethylammonium salt. The resulting budesonide 21-sulfate triethylammonium salt was added to the 10% sodium chloride solution while stirring. The resulting precipitate was filtered and the filtrate was purified by silica gel open column using chloroform / methanol (7/3) solution as eluent to yield 0.17 g (31% yield) of the title compound.

IR (cm ^-1 , Newsol): 1675 (C = O), 1640 (C = C), 1257 (S = O), 1035 (S = O)

¹ H-NMR (D ₂ O): 0.79 (s, 3H, C-18), 1.36 (s, 3H, C-19), 4.80 to 4.90 (AB q, 2H, C-21), 5.92 (s, 1H, C-4), 6.15 (d, 1H, C-1), 7.34 (d, 1H, C-2)

Elemental Analysis (C ₂₅ H ₃₃ SO ₉ Na): Calculated (C; 56.39, H; 6.20, S; 6.02), Found (C; 56.58, H; 6.10, S; 6.25)

Experimental Example 1: Measurement of apparent partition coefficient of corticosteroid 21-sulfate sodium derivative

To 10 ml of 0.1 M isotonic phosphate buffer (pH 6.8) in which a corticosteroid 21-sulfate sodium derivative (100 µg / ml each is a corticosteroid) was dissolved, 10 ml of 1-octanol previously saturated with the same buffer was added at 37 ° C. It was left for 24 hours with shaking. The amount of corticosteroid 21-sulfate sodium derivative remaining in the aqueous layer was then purified by HPLC [mobile phase: acetonitrile / 0.067 M, pH 4.5 phosphate buffer (4/6); Flow rate: 1.5 ml / min; Column: Waters (Waters μ bondapak C ₁₈ , 4.5 × 250, 5 μm); Detector: UV 248 nm]. The apparent partition coefficient was calculated by applying the formula (C ₀ -C _W ) / C _W (where C _O and C _W are the drug concentrations in the aqueous layer at initial and equilibrium, respectively). The measured results are shown in Table 1 below.

Sample data Dexa-s DS PD-S PD PC ^a 0.27 52.5 0.11 21.9 Solubility ^b (mg / ml) 14.4 - 46.2 -

A) ^a : 37 ° C., 1-octanol and pH 6.8 phosphate buffer as solvent system.

^b : Measured at 37 ° C., pH 6.8 phosphate buffer.

From the results shown in Table 1, all of the corticosteroid prodrugs of Formula 1 according to the present invention exhibited values of about 1/200 lower than those of corticosteroids, so their absorption in the intestine is estimated to be lower than that of corticosteroids. It became.

Experimental Example 2: Experiment of Prodrug Activation by Intestinal Contents in vitro )

Stomach, proximal small intestine (PSI), and distal small (DSI) of rats diluted 10-fold with Dexa-S or PD-S (100 μg equivalents as DS or PD, respectively) in isotonic phosphate buffer (pH 6.8), respectively Intestine (low intestine), cecum (cecum) and colon (colon) contents were mixed with 1.0 ml and incubated at 37 ℃, nitrogen. Take 100 μl of the culture at regular intervals, add 900 μl of methanol, centrifuge at 10,000 g, and then take 20 μl of the supernatant, using HPLC [mobile phase: acetonitrile / 0.067 M, pH 4.5 phosphate buffer (4/6); Flow rate: 1.5 ml / min; Column: Waters (Waters μ bondapak C ₁₈ , 4.5 × 250, 5 μm); Detector: UV 248 nm] to determine the release rate of DS or PD over time. The measured results are shown in Figs. 1, 2 and 3, respectively (in each case, the data shows the mean ± standard error of five experiments).

As shown in FIG. 1, after 8 and 10 hours of incubation with the cecal contents, the DS (●) released from Dexa-S reached 70% and 82% of the dose, and the amount of DS produced was This was consistent with the decreased amount of Dexa-S. When incubated with colon contents, DS (▼) liberated from Dexa-S at 8 and 10 hours elapsed to 19% and 26% of the dose, and the stomach, upper or lower intestine contents (■). DS was not liberated when incubated.

As shown in FIG. 2, the amount of PD generated after 6 hours of incubation with the cecal contents reached 29% of the amount of PD-S used for the culture, but after 8 hours after the amount of PD began to decrease. It was about 20%. In FIG. 2, ▲ indicates the amount of remaining PD-S, ▼ indicates the amount of reduced PD-S therefrom, and indicates the amount of PD measured. The amount of PD measured was less than the amount of reduced PD-S, which appeared to be greater with time. The decrease in PD was presumed to be due to the metabolism of PD in the culture, and the decrease in PD was observed when the PD was incubated with the cecal contents. That is, PD-S is hydrolyzed by sulfatase secreted from intestinal microorganisms in the intestine to produce PD, but the amount of PD produced by the metabolic process of reducing the steroid backbone of PD or PD-S It is estimated that it is less than the amount of reduced PD-S. On the other hand, PD was not detected from the culture medium with the stomach, upper small intestine, or the small intestine contents (■).

FIG. 3 shows dexamethasone (●) with time when cultured at 1.0 ° C. with 1.0 ml of cecal contents diluted 10-fold with Dexa-S or PD-S (100 μg equivalents of dexamethasone or prednisolone, respectively) with pH 6.8 isotonic phosphate buffer. Or graph comparing the release rate of prednisolone (■). Unlike PD-S, Dexa-S was converted to DS almost all after 12 hours.

When Dexa-S and PD-S were incubated with PSI or DSI contents, no DS or PD were released, so the corticosteroid 21-sulfate sodium derivatives of Formula 1 of the present invention were stable in the upper digestive tract and It was confirmed that it is mainly activated. Accordingly, the prodrugs of the present invention were expected to be activated at the colon site of human, which is the site corresponding to the cecum of the rat, to show efficacy.

Experimental Example 3: Measurement of the activation level of prodrugs by each site of the organ after oral administration of Dexa-S ( in vivo )

Dexa-S (1 mg equivalent / kg as DS) was orally administered to Sprague-Dawley rats weighing 250 to 280 g, and the animals were sacrificed at regular intervals before the contents of each organ. Dexa-S and DS from the amount was measured using HPLC in the same manner as in Experimental Example 2. The results are shown in FIGS. 4 and 5 (in each case, the data are shown as mean ± standard error of 4 experiments).

Figure 4 is a graph showing the amount of Dexa-S recovered from the contents of each organ part of the rat orally administered Dexa-S over time. 4, Dexa-S reached the highest concentration in the small intestine (◆) within 30 minutes and reached the highest concentration in the lower intestine (■) after 1.5 hours, and cecum (●) and colon (▼) with time. It can be seen that the drug is delivered.

Figure 5 is a graph showing the amount of DS recovered from the contents of each organ part of the rat orally administered Dexa-S over time. As shown in FIG. 5, almost no DS was detected in the upper and lower small intestine (■) over the entire time zone, indicating that Dexa-S is not activated and stable in the upper digestive tract. On the other hand, about 3 hours after drug administration, the amount of DS produced from Dexa-S began to increase in the cecum (●) and colon contents (▼), reaching a peak concentration at 6-8 hours, and then decreasing. Rarely was detected. It was then estimated to be excreted via the distal end of the colon.

Experimental Example 4: Determination of the concentration of DS recovered from the contents of each intestine after oral administration of DS ( in vivo )

DS (1 mg / kg) was orally administered to Sprague-Dawley rats weighing 250 to 280 g, and animals were sacrificed at regular time intervals. It was measured using HPLC by the method. The results are shown in FIG. 6 (in each case, the data are shown as mean ± standard error of 4 experiments).

As shown in Figure 6, DS reached the highest concentration in the upper small intestine (◆) after about 1.5 hours and reached the highest concentration in the lower intestine (■) at 4.5 time. However, in the cecum (●) and colon (▼), the recovery rate of drugs was very low compared to the small intestine. These results show that only a few reach the cecum or colon because DS is absorbed mostly in the upper digestive tract.

Experimental Example 5: Determination of cumulative concentration over time of DS recovered from small intestine or colon contents after oral administration of DS and Dexa-S ( in vivo )

After oral administration of DS and Dexa-S (1 mg / kg as DS) to rats, the animals were sacrificed at regular intervals and the cumulative concentrations of DS recovered from the small and large intestine contents were measured at different times. The results are shown in Fig. 7 (small intestine) and Fig. 8 (large intestine). 7 and 8 shows that DS is recovered only in the small intestine (●) and not detected in the large intestine, but when Dexa-S is administered (■), DS is not detected in the small intestine and DS is recovered only in the large intestine. Can be. This is because the amount of drug delivered to the large intestine is extremely limited because most of the drug is absorbed by the upper digestive tract when DS is administered, whereas intestinal microorganisms are mostly delivered to the large intestine because Dexa-S is stable and does not occur in the upper digestive tract. Hydrolysis by sulfatase secreted from means frees DS. Therefore, administration of the colon-targeting prodrug Dexa-S can be expected to be much more effective because the effective drug is mostly released from the affected colon.

Experimental Example 6: Measurement of the activation level of prodrugs by intestine after oral administration of PD-S in vivo )

PD-S (50 mg equivalent / kg) was administered orally to Sprague-Dawley rats weighing 250-280 g, and the animals were sacrificed at regular intervals before the amount of PD-S and PD from the contents of each organ. Was measured using HPLC in the same manner as in Experimental Example 2. The amounts of PD-S (●) and PD (■) recovered from the contents of the small intestine and the contents of the large intestine are shown in Figs. 9 (small intestine) and 10 (large intestine), respectively. Standard error).

As shown in FIG. 9, after oral administration of PD-S, the concentration of PD-S in the small intestine reached a maximum after 1.5 to 2 hours, and then rapidly decreased to be hardly detected after 6 hours. It is shown. No PD was detected from the contents of the small intestine over the entire time zone, indicating that the prodrug PD-S is stable in the small intestine.

As shown in FIG. 10, the concentration of PD-S in the colon contents reached a maximum between 2 and 4 hours, and then decreased slightly, but was present in a significant amount even about 6 hours. After 2 hours, PD was detected at a very low concentration and reached a peak at about 5 hours, but then decreased again. The concentration of recovered PD was very low over the entire time period. These results indicate that PD-S undergoes other metabolic processes in addition to hydrolysis in the intestine, as mentioned in Experimental Example 2.

Experimental Example 7 Measurement of Blood Levels of DS after Oral Administration of DS or Dexa-S

DS and Dexa-S (1 mg / kg as DS) were orally administered to Sprague-Dawley rats weighing 250 to 280 g, and then blood DS concentrations were measured at regular intervals. The results are shown in FIG. As shown in FIG. 11, when oral administration of DS (●), the blood concentration increased within 30 minutes, reached a maximum between 3 and 4 hours, and then rapidly decreased and hardly detected after 6 hours. In the case of oral administration of Dexa-S (■), neither DS nor Dexa-S were detected in the blood. This indicates that Dexa-S is not well absorbed from the digestive tract, and the amount of DS released from Dexa-S in the large intestine is below the detection limit over the entire time frame. Therefore, when DS is administered, the amount of DS absorbed is high, so side effects are concerned. However, administration of the colon-targeting prodrug Dexa-S does not need to worry about side effects.

Experimental Example 8: Determination of the concentration of DS or Dexa-S recovered from feces and urine after oral administration of DS or Dexa-S

Colonic target prodrugs intended for topical action are of great importance in reaching the colon without the drug being absorbed from the upper digestive tract. However, fast excretion drugs are difficult to measure the extent of absorption based on blood concentration alone. Therefore, in this experiment, DS or Dexa-S (1 mg equivalent / kg as DS) was orally administered to 250-280 g of Sprague-Dawley rats, which were fasted with water for 12 hours before the start of the experiment. The concentrations of Dexa-S and DS recovered from feces and urine collected for 24 hours were measured using HPLC in the same manner as in Experimental Example 2. The results are shown in FIG. 12 (in each case, the data are shown as mean ± standard error of 4 experiments).

As shown in FIG. 12, the DS administration group (■) had no DS recovered from feces and the amount DS recovered from urine was 260 μg. This result suggests that since DS is mostly absorbed in the upper digestive tract, it is not detected in feces and the absorbed DS is excreted mainly in urine. In the Dexa-S administration group (□), DS recovered from feces was 12 µg, and urine recovered 35 µg. Dexa-S was not detected at all in urine or feces. This result suggests that Dexa-S is not absorbed by the digestive tract and is converted almost entirely into DS until it is excreted into the feces. Considering the recovery rate in feces and urine as described above, Dexa-S reaches the large intestine without being absorbed in the upper digestive tract and liberates DS by the microorganisms, and it is applied to the large intestine area compared to the case where an equivalent amount of DS is administered. The high drug concentration, while the systemic absorption of DS is extremely limited, it can be seen that it is an excellent colon target prodrug that can minimize the systemic side effects caused by DS.

The corticosteroid 21-sulfate sodium derivative according to the present invention can effectively treat inflammatory bowel disease without side effects as a colon target prodrug of corticosteroids.

FIG. 1 shows the cecum (●), colon (▲), small intestine (PSI) or rats diluted 10-fold with dexamethasone 21-sulfate sodium (Dexa-S) (100 μg equivalent as dexamethasone) in pH 6.8 isotonic phosphate buffer. It is a graph showing the release rate of dexamethasone over time when dissolved in 1.0 ml of the small intestine (DSI) contents and cultured at 37 ℃;

FIG. 2 shows the cecum ()), small intestine (PSI), or small intestine (DSI) of rats diluted 10-fold with prednisolone 21-sulfate sodium (PD-S) (100 μg equivalent as prednisolone) with pH 6.8 isotonic phosphate buffer. It is a graph showing the release rate of prednisolone over time when incubated at 1.0 ml and 37 ℃ of the contents of [■; Amount of PD-S remaining in culture with cecal contents, ▲; Reduced amount of PD-S];

FIG. 3 shows the time-lapse when dexamethasone 21-sulfate sodium or prednisolone 21-sulfate sodium (100 μg equivalent, respectively, as dexamethasone or prednisolone) was diluted 10-fold with pH 6.8 isotonic phosphate buffer and 37 ml of cecal contents. Is a graph comparing the release rate of dexamethasone (●) or prednisolone (■) according to;

4 is a graph showing the concentration of dexamethasone 21-sulfate sodium recovered from the contents of each part of the digestive tract after sacrificing at regular intervals after oral administration of dexamethasone 21-sulfate sodium (1 mg equivalent / kg as dexamethasone) to rats. (◆; upper intestine, ■; lower intestine, ●; cecum, ▼; colon);

Figure 5 is a graph showing the concentration of dexamethasone recovered from the contents of each part of the digestive tract after sacrificing at regular time intervals after oral administration of dexamethasone 21-sulfate sodium (1 mg equivalent / kg as dexamethasone) to rats (■; small intestine) Upper or lower intestine, caecum, ▼ colon;

Figure 6 is a graph showing the concentration of dexamethasone recovered from the contents of each part of the digestive tract after sacrifice oral administration of dexamethasone (1 mg / kg) to rats (◆; small intestine, ■; small intestine, Cecum, ▼ colon);

FIG. 7 shows the cumulative concentrations of dexamethasone with oral administration of dexamethasone (•) or dexamethasone 21-sulfate sodium (■) at regular time intervals after sacrifice at regular time intervals (1 mg equivalent / kg as dexamethasone). Graph shown;

Figure 8 shows the cumulative concentrations of dexamethasone (*) or dexamethasone 21-sulfate sodium (■) to rats after oral administration (1 mg equivalent / kg as dexamethasone) at regular time intervals and recovered from the large intestine. Graph shown;

Figure 9 shows the concentrations of prednisolone (■) or prednisolone 21-sulfate sodium (-) recovered from the small intestine after sacrifice at regular intervals after oral administration of prednisolone 21-sulfate sodium (50 mg equivalent / kg as prednisolone) to rats. Is a graph showing;

Figure 10 shows the concentrations of prednisolone 21 or sulfate prednisolone (-) or prednisolone 21-sulfate (-) recovered from colon contents after oral administration of prednisolone 21-sulfate sodium (50 mg equivalent / kg as prednisolone) to rats at regular intervals. Is a graph showing;

11 is a graph showing the blood concentration of dexamethasone over time after oral administration of dexamethasone (•) or dexamethasone 21-sulfate sodium (■) to rats (1 mg equivalent / kg as dexamethasone);

Figure 12 shows the amount of dexamethasone recovered from urine and feces collected orally with dexamethasone (■) or dexamethasone 21-sulfate sodium (□) for 5 days orally (1 mg equivalent / kg / day as dexamethasone). This is a graph.

Claims (5)

Corticosteroid 21-sulfate sodium derivative of formula 1: [Formula 1] Where Represents a single or double bond, R ¹ represents hydrogen or CH ₃ , R ² represents hydrogen or F, R ³ represents ═O or OH, R ⁴ represents hydrogen, OH or CH ₃ , R ⁵ represents OH, or R ⁴ and R ⁵ together Indicates Except for hydrocortisone 21-sulfate sodium and cortisone 21-sulfate sodium. The method of claim 1, Is a double bond, R ¹ is H, R ² is H or F, R ³ is OH, R ⁴ is H or α-CH ₃ , and R ⁵ is OH derivative. a) reacting a corticosteroid of formula (2) with sulfur trioxide triethylamine of formula (3) to obtain a corticosteroid 21-sulfate triethylammonium salt of formula (4), and then reacting it with a saturated saline solution Method for preparing corticosteroid 21-sulfate sodium derivative: [Formula 2] [Formula 3] _{SO 3 · N (C 2 H} 5) 3 [Formula 4] [Formula 1] Where , R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined in claim 1. Pharmaceutical composition for the prevention or treatment of inflammatory bowel disease containing corticosteroid 21-sulfate sodium derivative of formula 1 as an active ingredient: [Formula 1] Where Represents a single or double bond, R ¹ represents hydrogen or CH ₃ , R ² represents hydrogen or F, R ³ represents ═O or OH, R ⁴ represents hydrogen, OH or CH ₃ , R ⁵ represents OH, or R ⁴ and R ⁵ together Indicates. The composition according to claim 4, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease.