TW202003045A - Lactulose glycoside compound, preparation method thereof and medicinal use thereof for providing the function of colon-localized drug release to prevent or treat the intestinal diseases - Google Patents

Abstract

The present invention relates to the field of medicinal chemistry and, in particular, to a compound represented by formula (I), a preparation method thereof, and a medicinal use thereof. In the compound represented by the formula (I), the lactulose group is connected to a heteroatom in aglycon (G) through a glycoside bond. The aglycon (G) is a group formed by removing a hydrogen atom from a heteroatom of an active drug molecule and, as represented by the zigzag-like symbol, the lactulose group is connected to a heteroatom in the aglycon (G) through an <alpha>-glycoside bond or a <beta>-glycoside bond. The pharmacokinetic test proves that the lactulose glycoside compound according to the present invention may pass through the mammalian gastrointestinal tract without being significantly absorbed by the gastrointestinal tract and without being significantly hydrolyzed by endogenous enzymes of the mammalian host, so that the lactulose glycoside compound may reach the mammalian colon portion to release the active drug in the colon under the action of the colonic flora. The lactulose glycoside compound has the function of colon-localized drug release for preventing or treating the intestinal diseases.

Description

Lactulose glycoside compound, its preparation method and use

The invention relates to the field of medicinal chemistry, in particular to a lactulose glycoside compound with colonic localized delivery and release, its preparation method and its medical use.

Most oral drugs are absorbed through the upper digestive tract (stomach, small intestine) and enter the systemic circulation to play a therapeutic role. For diseases existing in the lower end of the digestive tract (cecum, colon, rectum), it is difficult for the drug to directly reach the lesion site, but only depends on the concentration of the drug circulating in the system to exert the effect. Such colon diseases include inflammatory bowel disease (ulcerative colitis, Klein disease, etc.), intestinal infection (amebic bowel disease), bowel cancer (colon cancer, rectal cancer), irritable bowel syndrome, chronic Constipation, etc. For the treatment of these diseases, most commonly used drugs such as glucocorticoids, non-steroidal anti-inflammatory drugs, immunomodulators, monoclonal antibodies, broad-spectrum cytotoxic drugs, etc. are mostly administered orally or by injection, and are exposed through the systemic system of drugs Amount to treat local disease. The obvious shortcomings are the unnecessary systemic side effects when the drug concentration is reached locally, such as the treatment of ulcerative colitis, the effect of glucocorticoid treatment on the hypothalamus-pituitary-adrenal gland, and the use of immunomodulators It leads to abnormal and disordered systemic immune function. The most ideal way of administration is local administration in the colon, which can increase the local drug concentration, avoid or reduce the systemic exposure of the drug, and thus reduce systemic side effects.

In order to solve the above problems, the researchers provided different colon-targeted drug solutions. For example, balsalazide releases the active drug 5-aminosalicylic acid through the colon microflora azo reductase to exert an anti-inflammatory effect. Mesalazine enema and budesonide enema are directly used for enema treatment at the end of the digestive tract (rectal and sigmoid colon). Colon-targeted preparations prepared with various complex preparation techniques are also frequently reported. In addition, there are reports of using prodrug technology to connect the active drug with macromolecular carriers (dextran, xylan, cyclodextrin, etc.), hoping to release the active drug after delivering the drug to the colon. For details, please refer to the following documents: Chinese Pharmacology Bulletin, (2002), 18(3), 328-330; PLA Pharmaceutical Journal, (2001), 17(2), 62-64; Carbohydrate Polymers (2017), 157, 1442-1450; and Journal of Pharmaceutical Sciences (2001), 90(12), 2103-2112.

Therefore, there is a great need to develop a simple small molecule prodrug, which has a single molecular structure and precise composition, and this prodrug is an active drug connected with a small molecule carrier, which is not absorbed by the upper digestive tract or absorbed little, and is not endogenous Sexual enzymes are degraded, and most drugs can be positioned and delivered to the colon, and then the active drugs are quickly released in the colon to play a therapeutic effect.

The present invention has been implemented for the above purposes, and in particular, the present invention discloses a class of lactulose glycoside compounds that pass through the gastrointestinal tract of mammals without significant absorption by the gastrointestinal tract and are not significantly hydrolyzed by endogenous enzymes of the mammalian host. Lactulose glycoside compounds can reach the colon of mammals and release active drugs in the colon under the action of colonic flora. It has the function of colon-specific drug release and is used for the prevention or treatment of intestinal diseases.

Lactulose, also known as lactulose, lactulose, and galactosyl fructoside, is a synthetic disaccharide composed of galactose and fructose bonded by β-1,4 glycosidic bonds. Lactulose has a good bifidobacterium proliferation function, has the effect of regulating the intestinal microecological balance, and has a wide range of applications in medicine, food and animal feeding. Lactulose can reduce blood ammonia, catharsis, treat constipation, and is also used for the prevention and treatment of various liver diseases. It can also be used as a low-calorie sweetener and functional food additive in food. It is widely used. However, as of now, there has been no report on the use of lactulose as a small molecule carrier to locate and deliver active drugs.

其中，乳果糖基通過糖苷鍵與苷元(G)中的雜原子連接，所述苷元(G)為從活性藥物分子的雜原子除去一個氫原子所形成的基團，並且「

」表示乳果糖基通 過α糖苷鍵或β糖苷鍵與苷元(G)中的雜原子連接。 According to one aspect of the present invention, the present invention discloses a lactulose glycoside prodrug compound, which is represented by the following formula (I):

Among them, the lactulose group is connected to the heteroatom in the aglycon (G) through a glycosidic bond, which is a group formed by removing one hydrogen atom from the heteroatom of the active drug molecule, and "

"Indicates that the lactulose group is connected to the heteroatom in the aglycon (G) through an alpha glycosidic bond or a beta glycosidic bond.

」表示乳果糖基通過α糖苷鍵或β糖苷鍵與苷元(G)中的雜原子連接。即，所形成的乳果糖糖苷前藥化合物具有由下式(III)或(IV)表示的結構：

in the text,"

"Indicates that the lactulose group is connected to the heteroatom in the aglycon (G) through an alpha glycosidic bond or a beta glycosidic bond. That is, the formed lactulose glycoside prodrug compound has a structure represented by the following formula (III) or (IV):

The lactulose glycoside prodrug compound according to the present invention may be an alpha glycosidic bond compound, a beta glycosidic bond compound, or a mixture of the two.

According to the present invention, the term "aglycon" has a common meaning in the art, that is, a non-sugar portion condensed with a sugar in a glycoside compound.

The active drug according to the present invention is a chemical component that has a certain pharmacological activity and is used for the prevention or treatment of human diseases. The active drug may be a drug approved for marketing in various countries, or may be a candidate drug in the stage of human clinical trials.

According to the technical solution of the present invention, the heteroatom system is selected from any one of an oxygen atom, a nitrogen atom and a sulfur atom. And the aglycon (G) is a group formed by removing one hydrogen atom from the alcoholic hydroxyl group, phenolic hydroxyl group, amino group, amide group or heterocyclic ring NH in the active drug molecule.

According to the technical content of the present invention, the case in which two or more lactulose groups are linked by aglycon is also within the scope of the present invention.

According to some embodiments of the present invention, the active drug molecule system is selected from the group consisting of glucocorticoids, Apremilast, Prucalopride, Mesalamine, Metronidazole (Metronidazole), azathioprine (Azathioprine), 6-mercaptopurine (6-Mercaptopurine), 5-fluorouracil (5-Fluorouracil) and tofacitinib (Tofacitinib) any one.

The glucocorticoid drugs refer to a class of synthetic drugs that have similar effects to the glucocorticoids secreted by the adrenal cortex in the human body. They have the role of regulating the biosynthesis and metabolism of sugar, fat and protein, and also have anti-inflammatory properties. effect. Glucocorticoid drugs include not only endogenous substances with the above characteristics and activities, but also many synthetic drugs with similar structures and activities that have been optimized in structure. Examples of glucocorticoids include, but are not limited to: cortisone acetate, prednisone, prednisone acetate, hydrocortisone, hydrocortisone acetate, hydrocortisone butyrate, hydrocortisone succinic acid Sodium, hydrocortisone sodium phosphate, fludrocortisone, fludrocortisone acetate, prednisolone, prednisolone acetate, prednisolone sodium succinate, prednisolone sodium phosphate, beta Betamethasone, Betamethasone Acetate, Betamethasone Dipropionate, Clobetasol Butyrate, Betamethasone Valerate, Betamethasone Sodium Phosphate, Beclomethasone Dipropionate, Mometasone Furoate, Clobetasol Propionate Cable, dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, p-fluoromethasone, fluticasone propionate, methylprednisolone, methylprednisolone acetate, flumiron, triamcinolone, triamcinolone, triamcinolone acetonide Annide, Flusinide, Ansinide, Hasinaide, Sononide, Budesonide, and Decote.

The glucocorticoid drugs are preferably selected from Hydrocortisone (Hydrocortisone), Prednisolone (Prednisolone), Methylprednisolone (Methylprednisolone), Dexamethasone (Dexamethasone), Betamethasone (Betamethasone), Qu Ansiron (Triamcinolone) and budesonide (Budesonide).

The active drug is preferably selected from the group consisting of prednisolone, dexamethasone, budesonide, aprost, procapril and mesalazine.

其中，R₁為H或F；R₂為H、F或-CH₃；R₃為-OH；並且R₄為H、-OH或-CH₃； 或者R₃和R₄連接成基團

、

或

。 In the formula (I) shown above, the aglycon (G) is a group represented by the following formula (II):

Where R ₁ is H or F; R ₂ is H, F or -CH ₃ ; R ₃ is -OH; and R ₄ is H, -OH or -CH ₃ ; or R ₃ and R _{4 are} connected to form a group

,

or

.

其中Me表示-CH₃。 Preferably, the aglycone (G) is selected from any one of the following:

Where Me represents -CH ₃ .

其中Me表示-CH₃。 Preferably, the aglycone (G) is selected from any one of the following:

Where Me represents -CH ₃ .

其中Me表示-CH₃。 More preferably, the aglycon (G) is selected from any one of the following:

Where Me represents -CH ₃ .

More specifically, the compound is any one selected from the group consisting of (11β)-11,17-dihydroxy-21-{[4-O-(β-D-galactopyranosyl)-D -Fructofuranosyl]oxy}pregnant-1,4-diene-3,20-dione; (11β)-11,17-dihydroxy-21-{[4-O-(β-D-pyran Galactosyl)-D-fructofuranosyl]oxy}pregnant-4-ene-3,20-dione; (6α,11β)-11,17-dihydroxy-6-methyl-21-{[4 -O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1,4-diene-3,20-dione; (11β,16α)-9-fluoro- 11,17-Dihydroxy-16-methyl-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1,4-diene- 3,20-diketone; (11β,16β)-9-fluoro-11,17-dihydroxy-16-methyl-21-{[4-O-(β-D-galactopyranosyl)-D -Fructofuranosyl]oxy}pregnant-1,4-diene-3,20-dione; (11β,16α)-9-fluoro-11,16,17-trihydroxy-21-{[4-O -(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}progesterone-1,4-diene-3,20-dione; (11β,16α)-11-hydroxy-16, 17-(butylene dioxy)-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1,4-diene-3,20 -Diketone; 2-methyl-5-nitro-1-{2-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}ethyl}imidazole; 6-[(1-methyl-4-nitro-1 H -imidazol-5-yl)sulfur]-9-[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl ]-9 H -purine; 6-mercapto-9-[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]-9 H -purine; 5-fluoro-1-[4 -O-(β-D-galactopyranosyl)-D-fructofuranosyl]pyrimidine-2,4(1H,3H)-dione; N -{2-[(S)-1-(3- Ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl]-1,3-dioxo-2,3-dihydro-1 H -isoindole-4- Group}- N' -[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]acetamide; 4-amino-5-chloro-2,3-dihydro- N- [1-(3-Methoxypropyl)-4-pyridinyl]- N' -[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]-7-benzene Pyrofurancarboxamide; 5-chloro-2,3-dihydro-4-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]amino}- N -[1 -(3-methoxypropyl) -4-pyridyl]-7-benzofurancarboxamide; 5-amino-2-{[4-O-(β-D-galactopyranosyl)-D-fruranose]oxy}benzene Formic acid; 2-hydroxy-5-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]amino}benzoic acid; 17-hydroxy-21-{[4-O-( β-D-galactopyranosyl)-D-fructofuranosyl]oxy}progesterone-1,4-diene-3,11,20-trione; and 4-{ N -methyl- N'- [(3R,4R)-4-methyl-1-(2-cyanoacetamide)pyridin-3-yl]amino}7-[4-O-(β-D-galactopyranosyl) -D-fructofuranosyl]-7 H -pyrrolo[2,3- d ]pyrimidine.

According to a second aspect of the present invention, there is provided a pharmaceutical composition containing the lactulose glycoside prodrug compound as described above and a pharmaceutically acceptable carrier.

According to the third party of the present invention, the use of the compound as described above for the preparation of a medicament for preventing or treating intestinal diseases, wherein the intestinal diseases are selected from ulcerative colitis, Crohn's disease, infectious enteritis , Irritable bowel syndrome, chronic constipation, amoebic bowel disease, colon cancer and rectal cancer, and more preferably, the intestinal disease is selected from ulcerative colitis, Crohn's disease and chronic constipation.

As demonstrated in the appended examples, the lactulose glycoside compound according to the present invention can pass through the mammalian gastrointestinal tract without significant absorption by the gastrointestinal tract and is not significantly hydrolyzed by endogenous enzymes of the mammalian host, so the lactulose glycoside compound can Reaching the site of the mammal's colon, the active drug is released in the colon under the action of the colon flora. The lactulose glycoside compound has the function of colon-specific drug release, and can be used for the prevention or treatment of intestinal diseases.

The embodiments of the present invention will be further described below with reference to the drawings. The examples listed below are used to clarify the inventive features and applications of the present invention, rather than to limit the scope of the present invention. Anyone who is familiar with this skill will not deviate from it. Within the spirit and scope of the present invention, some changes and modifications can be made, so the scope of protection of the present invention shall be subject to the scope defined in the appended patent application.

The following drawings are provided to describe in detail the embodiments according to the present invention, in which: FIG. 1 shows the change in the concentration of prednisolone in the contents of the first 1/3 segment of the colon (including the cecum) of rats after intragastric administration; The concentration change of prednisolone in the content of 2/3 of the colon of rats after intragastric administration; Figure 3 shows the blood concentration-time curve of prednisolone in different groups of rats after intragastric administration; Figure 4 shows the weight change curve of rats in different groups after intragastric administration; Figure 5 shows the comparison of body weight changes of different groups of rats on day 8 of modeling; Figure 6 shows the different groups of rats on day 8 of modeling Comparison of the colonic coefficients; and Figure 7 shows the comparison of histopathological score results of different groups of rats on the 8th day of modeling.

Specifically, the example compounds provided by the present invention are selected from the structures shown in Table 1 below.

Next, a method for preparing the compound of the present invention will be described.

The compound of the present invention can be prepared, for example, by the following preparation method.

(Preparation method A)

In pyridine (Py) solvent, in the presence of dichloromethane (CH ₂ Cl ₂ ), lactulose is reacted with benzoyl chloride (BzCl) to obtain octyl benzoyl lactulose (A). Then, under the action of trimethylsilyl triflate (abbreviated as TMSOTf), it undergoes glycosylation condensation reaction with the primary hydroxyl group at position 21 of the glucocorticoid drug (B), and is purified by column chromatography to obtain an intermediate ( C) Finally, the benzoyl group on the sugar group is removed by ammonia methanol solution to obtain the target product (D), and its typical examples are compounds 1 to 7 shown in the above table. Similarly, the octanoyl lactulose (A) and metronidazole are glycosylated, and then the benzoyl group is removed to obtain the target compound 8. Drug molecules with primary hydroxyl groups can be used to prepare corresponding target compounds using the above method.

(Preparation method B)

Lactulose is acetyl anhydride/pyridine acetylated to obtain all octyl acetyl lactulose (Compound F).

In acetonitrile solvent, azathioprine, 6-mercaptopurine, 5-fluorouracil (5-Fu) or Alpster are silanized with N,O-bis(trimethylsilyl)acetamide (abbreviated as BSA) , Under the action of trimethylsilyl trifluoromethanesulfonate (TMSOTf), and glycoside condensation reaction with octadecyl lactulose (F), purified by column chromatography to obtain intermediate (G), and then Alkali treatment removes the acetyl group on the sugar group and prepares and purifies to obtain the target product (H) (compounds 9, 10, 11, 12).

Tofacitinib (Tofacitinib) can be used to obtain the corresponding lactulose glycoside (Compound 18) using a synthetic method similar to the above.

(Preparation method C)

Octabenyl lactulose (A) is converted to chlorinated compound (K) by hydrogen chloride or hydrogen bromide. Procabulide reacts with chlorinated compound (K) to form intermediate (M). The crude product is purified by column chromatography , And then subjected to alkali treatment to remove the benzoyl group on the sugar group and preparation and purification to obtain compound 13.

Prucalopride is silanized with N,O-bis(trimethylsilyl)acetamide (abbreviated as BSA) under the action of tert-butyldimethylsilyl trifluoromethanesulfonate (TBSOTf) , Glycosylation condensation reaction with octanoyl lactulose (A), purified by column chromatography to obtain intermediate (N), and then treated with alkali to remove the benzoyl group on the sugar group and preparation and purification to obtain the compound 14.

(Preparation method D)

5-Aminosalicylic acid (mesalazine) is methylated to obtain mesalamine methyl ester (P), and the compound (Q) is obtained by protecting the amino group with benzyl chloroformate. Glycosylation reaction of the phenolic hydroxyl group of compound (Q) with chlorinated sugar (K) to obtain mesalazine O-glycoside compound (R), and finally the benzyloxycarbonyl group is removed by hydrogenation, the methyl ester and benzoic acid are removed by alkaline hydrolysis Ester to give compound 15.

After mesalazine methyl ester (P) is protected by di-tert-butyl dicarbonate (Boc ₂ O), the amino group is reacted with benzyl chloride to protect the phenolic hydroxyl group, and then the Boc protecting group is removed to obtain compound (S). The amino group of compound (S) then undergoes glycosylation reaction with chlorosugar (K) to obtain mesalazine N-glycoside compound (T), and finally the benzyl group is removed by hydrogenation, and the methyl ester and benzoate are removed by alkaline hydrolysis , To give compound 16.

The compound of formula (I) of the present invention can be mixed with a pharmaceutically acceptable carrier to prepare various oral pharmaceutical compositions. After the compound of formula (I) is mixed with acceptable auxiliary additives such as disintegrants, excipients, lubricants, binders, fillers, etc., it can be made into tablets, pills and capsules according to conventional methods Agent or a variety of corresponding sustained-release agents, controlled-release agents and other solid dosage forms of oral drugs; with commonly used solubilizers, emulsifiers, wetting agents, foaming or defoaming agents and other surfactants, diluents, preservatives, stabilizers Agents, flavoring agents, thickeners, etc. are mixed, and can be made into oral preparations such as liquid preparations, syrups and other liquid preparations according to the corresponding conventional methods.

The invention also provides in vitro stability tests of the compounds of the examples and methods of drug metabolism and pharmacodynamic tests. The results show that the compound of the present invention has good colon-specific drug release and the advantages of treating intestinal diseases, and is particularly suitable for the drug treatment of the cecum and colon.

Therefore, the present invention also provides the use of a compound of formula (I) in the preparation of a pharmaceutical composition for the prevention or treatment of intestinal diseases or disorders selected from ulcerative colitis and Crohn Disease, infectious enteritis, irritable bowel syndrome, chronic constipation, amoebic bowel disease, colon cancer, rectal cancer.

The intestinal disease or condition is preferably selected from ulcerative colitis, Crohn's disease, chronic constipation.

Examples

The preparation method of the compound according to the present invention is specifically exemplified below through examples. It should be noted that the technical solution according to the present invention is not limited to the embodiments.

In the following examples, lactulose was purchased from Bellingway Technology (content 99.0%). Prednisone (content 99.0%), prednisolone (content 99%), hydrocortisone (content 98.8%), dexamethasone (content 98.8%), betamethasone (content 99.8%), triamcinolone (Content 99.0%), budesonide (content 98.5%) were purchased from Shandong Taihua Biotechnology Co., Ltd. Methylprednisolone (content 98%) was purchased from Wuhan Dongkangyuan Technology Co., Ltd., and metronidazole (content 98.0%) was purchased from Saen Chemical Technology Co., Ltd. Azathioprine (content 98%) was purchased from Aladdin reagent. 6-Mercaptopurine (content 98.0%) and 5-fluorouracil (98.0%) were purchased from Shanghai Bide Pharmaceutical Technology Co., Ltd. Apster (98.0% content) was purchased from Adamas Reagent Co., Ltd. Procabride (content 99.2%) was purchased from Zhiwei Chemical Technology Co., Ltd. Mesalazine (content 99.7%) was purchased from Hubei Xingyinhe Chemical Co., Ltd.

In this application, unless otherwise specified, other reagents used are commercially available.

In addition, in this specification, unless otherwise specified, "%" means% by weight.

Preparation Example 1 (11β)-11,17-dihydroxy-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1,4-diene-3 ,20-diketone (Compound 1)

step one:

Dissolve lactulose (10g, 0.029mol) in 100ml pyridine, add 60ml dichloromethane, stir and cool to 0°C, dropwise add benzoyl chloride (61.1g, 0.435mol), after the dropwise addition, raise to room temperature React for 16 hours.

The reaction solution was concentrated to dryness under reduced pressure, dissolved by adding 200ml of dichloromethane, washed with citric acid solution (pH>4) (120ml*3 times), saturated sodium bicarbonate (120ml*1 time), saturated sodium chloride The solution was washed (20 ml*1 time), the organic phase was dried with anhydrous sodium sulfate for 30 minutes, filtered, concentrated under reduced pressure and purified by column chromatography to obtain 30.0 g of white solid as the product of step 1 (compound A), yield 87.2 %. ESI(+) m/z: 1192.3 [M+NH ₄ ] ⁺ 1197.2 [M+Na] ⁺ .

Step two:

Under the protection of argon, the product of the first step (10.1g, 8.59mmol) and prednisolone (2.6g, 7.16mmol) were dissolved in 400ml of dichloromethane, 5g of 4A molecular sieve was added, stirring was started and the temperature was reduced to- At 35°C, TMSOTf (2.7g, 12.2mmol) was added dropwise. After maintaining the temperature for 2 hours, the temperature was raised to -15°C and the reaction was performed for 2-3 hours.

Trimethylpyridine (2.2g, 17.9mmol) was added dropwise to the reaction mixture, and the temperature was raised to 25°C and stirring was continued for 1 hour. Then the reaction solution was washed successively with citric acid solution (pH>4) (200ml*3 times), saturated sodium carbonate (200ml*1 time), saturated sodium chloride solution (200ml*1 time), and the organic phase was used Dry with anhydrous sodium sulfate for 30 minutes and filter. The filtrate was concentrated and purified by column chromatography to obtain 7.5g of white solid as the product of step two, with a yield of 61.5%. ESI(+) m/z: 1435.2 [M+Na] ⁺ .

Step 3: Preparation of Compound 1

The product of the second step (5.4g, 3.8mmol) was dissolved in 10ml of tetrahydrofuran, 40ml of ammonia methanol (10M) was added, and stirred at room temperature for 72 hours.

The reaction solution was concentrated to dryness under reduced pressure, dissolved in water, and then concentrated to dryness, and repeated three times. To the residue were added 30 ml of water and 10 ml of methyl tert-butyl ether. After dissolution, the liquid was separated. The aqueous phase was concentrated under reduced pressure, and the remaining water was removed with absolute ethanol. The resulting solid was suspended in 30 ml of acetone and stirred for 50 minutes, and filtered. The filter cake was rinsed with a small amount of acetone and drained. The filter cake was recrystallized and purified with methanol/water (v/v)=1/1 to obtain 1.5 g of a white solid as the title compound (Compound 1), with a yield of 57.7%. . ESI(+) m/z: 707.4 [M+Na] ⁺ .

¹ H-NMR (DMSO-d6,500M, BRUKER AV-500): δ (ppm) 7.32 (1H, d, =CH), 6.19 (1H, dd, J ₁ =1.5Hz, J ₂ =10Hz, =CH ), 5.91 (1H, s, =CH), 5.23 (1H, s, CH(O) ₂ -), 5.09 (1H, d, O-CH ₂ -CO-), 4.82-4.79 (1H, t), 4.74-4.71(2H,m), 4.69(1H,d,O-CH ₂ -CO-), 4.62(1H,s), 4.53(1H,d), 4.38(1H,d), 4.29-4.23(3H , m), 4.14-4.10 (2H, m), 3.84-3.81 (1H, m), 3.78-3.75 (1H, m), 3.61-3.59 (2H, m), 3.56-3.48 (3H, m), 3.46 -3.42(1H,m), 3.40-3.36(2H,m), 3.32-3.30(2H,m), 2.56-2.53(2H,m), 2.30-2.28(1H,m), 2.04-2.02(2H, m), 1.89-1.85 (1H, dd, J ₁ = 3Hz, J ₂ = 13.5Hz), 1.66-1.60 (3H, m), 1.41-1.39 (1H, m), 1.38 (3H, s, -CH ₃ ), 1.30-1.23(1H,m), 1.02-0.99(1H,m), 0.91-0.88(1H,dd,J ₁ =3Hz,J ₂ =10.5Hz), 0.779(3H,s,-CH ₃ ) .

Preparation Example 2 (11β)-11,17-dihydroxy-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-4-ene-3,20- Diketone (Compound 2)

Preparation Example 2 was carried out in a similar manner to Preparation Example 1, except that prednisolone was replaced with hydrocortisone to obtain 0.45 g of the title compound. ESI(+) m/z: 709.3 [M+Na] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 5.56 (1H, s, =CH), 5.29 (1H, s), 5.16 (1H, d), 4.87-4.78 (5H , m), 4.68-4.63 (1H, m), 4.46 (1H, m), 4.35-4.34 (1H, m), 4.29-4.22 (3H, m), 4.18-4.11 (2H, m), 3.82-3.80 (1H,m), 3.77-3.74 (1H,m), 3.59 (2H,m), 3.56-3.42 (4H,m), 3.40-3.34 (2H,m), 3.30 (2H,m), 2.57-2.54 (1H,m), 2.45-2.34(2H,m), 2.21-2.17(2H,m), 2.10-2.07(1H,m), 1.93-1.88(3H,m), 1.81-1.75(1H,m) 、1.66-1.55(3H,m)、1.42-1.40(1H,m)、1.36(3H,s,-CH ₃ )、1.28-1.24(1H,m)、1.06-0.98(1H,m)、0.88- 0.85(1H,d), 0.75(3H,s,-CH ₃ ).

Preparation Example 3 (6α,11β)-11,17-dihydroxy-6-methyl-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1 ,4-diene-3,20-dione (Compound 3)

Preparation Example 3 was carried out in a similar manner to Preparation Example 1, except that prednisolone was replaced with methylprednisolone to obtain 0.73 g of the title compound. ESI(+) m/z: 721.3 [M+Na] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ(ppm) 7.33(1H,d,=CH), 6.19(1H,d), 5.82(1H,s,=CH), 5.28( 1H,s), 5.15(1H,d), 4.89-4.86(1H,m), 4.83-4.78(3H,m), 4.67-4.62(1H,m,), 4.59(1H,s), 4.46(1H , d), 4.36-4.33 (1H, m), 4.27-4.22 (2H, m), 4.14-4.10 (2H, m), 3.81 (1H, m), 3.77 (1H, m), 3.61-3.59 (2H , m), 3.56-3.46 (3H, m), 3.43 (1H, m), 3.39-3.36 (2H, m), 3.29-3.25 (2H, m), 2.66-2.65 (1H, m), 2.57-2.53 (1H,m), 2.11-2.02(2H,m), 1.62-1.59(3H,m), 1.42(1H,m), 1.38(3H,s,-CH ₃ ), 1.31-1.29(1H,m) , 1.05-1.04 (3H, d, -CH ₃ ), 0.87-0.84 (1H, m), 0.77 (3H, s, -CH ₃ ), 0.72-0.66 (1H, m).

Preparation Example 4 (11β,16α)-9-fluoro-11,17-dihydroxy-16-methyl-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxygen} Progesterone-1,4-diene-3,20-dione (Compound 4)

Preparation Example 4 was carried out in a similar manner to Preparation Example 1, except that prednisolone was replaced with dexamethasone to obtain 0.95 g of the title compound. ESI(+) m/z: 739.3 [M+Na] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 7.30 (1H,d,=CH), 6.24 (1H,d,=CH), 6.01 (1H,s,=CH) , 5.32-5.27 (1H, m), 5.16 (1H, d), 5.08 (1H, s), 4.88-4.80 (4H, m), 4.72-4.64 (2H, m), 4.47 (1H, s), 4.36 (1H,m), 4.27-4.22(1H,m), 4.15-4.13(3H,m), 3.83-3.79(2H,m), 3.60-3.53(5H,m), 3.44(2H,m), 3.29 (2H,m), 2.92 (1H,m), 2.67-2.59 (1H,m), 2.39-2.31 (2H,m), 2.16-2.06 (2H,m), 1.76 (1H,m), 1.66-1.58 (1H,m), 1.52(1H,m), 1.49(3H,s,-CH ₃ ),1.36-1.34(1H,m), 1.06(1H,m), 0.87(3H,s,-CH ₃ ) , 0.79 (3H, d, -CH ₃ ).

Preparation Example 5 (11β,16β)-9-fluoro-11,17-dihydroxy-16-methyl-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxygen} Progesterone-1,4-diene-3,20-dione (Compound 5)

Preparation Example 5 was carried out in a similar manner to Preparation Example 1, except that prednisolone was replaced with betamethasone to obtain 0.58 g of the title compound. ESI(+) m/z: 739.3 [M+Na] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 7.30 (1H,d,=CH), 6.23 (1H,d,=CH), 6.01 (1H,s,=CH) , 5.24 (1H, s), 5.20 (1H, d), 5.14-5.12 (1H, d), 4.88-4.77 (4H, m), 4.60-4.55 (1H, m), 4.47 (1H, d), 4.40 -4.35(1H,m), 4.29-4.24(1H,m), 4.14-4.10(3H,m), 3.85-3.83(1H,m), 3.79-3.77(1H,m), 3.60-3.46(5H, m), 3.45-3.42 (1H, m), 3.40-3.36 (2H, m), 3.30 (2H, m), 2.66-2.60 (1H, m), 2.46-2.32 (2H, m), 2.09-2.07 ( 2H,m), 1.98-1.82(3H,m), 1.49(3H,s,-CH ₃ ),1.40-1.34(2H,m), 1.04-1.02(2H,m), 0.95(3H,s,- CH ₃ ).

Preparation Example 6 (11β,16α)-9-fluoro-11,16,17-trihydroxy-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}progesterone- 1,4-diene-3,20-dione (Compound 6)

Preparation Example 6 was carried out in a similar manner to Preparation Example 1, except that triamcinolone was replaced with prednisolone to obtain 0.38 g of the title compound. ESI(+) m/z: 741.3 [M+Na] ⁺ .

Preparation Example 7 (11β,16α)-11-hydroxy-16,17-(butylene dioxy)-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnancy Steroid-1,4-diene-3,20-dione (Compound 7)

Preparation Example 7 was carried out in a similar manner to Preparation Example 1, except that budesonide was substituted for prednisolone to obtain 1.2 g of the title compound. ESI(+) m/z: 777.3 [M+Na] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 7.32 (1H,d,=CH), 6.18 (1H,d,=CH), 5.92 (1H,s,=CH) , 5.19-5.17 (1H, m), 5.14 (1H, m), 5.05 (1H, d), 4.81-4.75 (4H, m), 4.71 (1H, m), 4.64-4.62 (1H, d), 4.59 -4.57(1H,m), 4.45(1H,m), 4.35(2H,m), 4.31-4.24(2H,m), 4.15-4.13(2H,m), 3.82-3.75(2H,m), 3.60 (3H,m), 3.57-3.53 (3H,m), 3.45-3.43 (3H,m), 3.40-3.37 (2H,m), 3.29 (2H,m), 2.77-2.67 (1H,m), 2.31 (1H,m), 2.09 (1H,s), 1.98 (2H,m), 1.79-1.72 (3H,m), 1.61-1.52 (4H,m), 1.42 (2H,m), 1.38 (3H,s ,-CH ₃ ), 1.35-1.23(2H,m), 1.15-1.11(1H,m), 1.07-1.04(3H,t,-CH3), 1.01-0.93(2H,m), 0.88((1H,m m), 0.86 (3H, s, -CH ₃ ), 0.81 (1H, s).

Preparation Example 8 2-methyl-5-nitro-1-{2-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}ethyl}imidazole (Compound 8)

Preparation Example 8 was carried out in a similar manner to Preparation Example 1, except that prednisolone was replaced with metronidazole to obtain 1.9 g of the title compound. ESI(+) m/z: 518.2 [M+Na] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 8.03 (1H, s, Ar-H), 5.06-5.04 (1H, d), 4.87-4.85 (1H, d), 4.82-4.79(1H,d), 4.76(2H,m), 4.47-4.43(3H,m), 4.35-4.32(1H,t), 4.09-4.07(1H,d), 3.94-3.91(1H,t ), 3.81-3.79(2H,t), 3.72-3.69(1H,m), 3.60(1H,m), 3.54-3.49(2H,m), 3.48-3.39(5H,m),3.34-3.27(3H ,m), 2.48(3H,s,-CH ₃ ).

Preparation Example 9 6-[(1-methyl-4-nitro-1H-imidazol-5-yl)sulfur]-9-[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl] -9H-purine (Compound 9)

step one:

At room temperature, add 10.0 g of lactulose (0.029 mol) to the reaction bottle, and then add 45.0 g of acetic anhydride (0.435 mol) and 34.4 g of pyridine (0.43 mol). After stirring for about 1 h, dissolve it. Stir at room temperature for 14 h. The acetic acid, pyridine and acetic anhydride were distilled off under reduced pressure, and the residue was purified by petroleum ether/ethyl acetate column chromatography to obtain octaacetyl lactulose (compound F), weighing 16.5 g of product, with a yield of 83.2%.

Step two:

Under argon atmosphere, mix azathioprine (1.5g, 5.4mmol), N, O-bis (trimethylsilyl) acetamide (BSA) (1.3g, 6.48mmol), acetonitrile (50ml) After heating to reflux for 3 hours, the temperature was lowered to 0°C, the above compound F (5.5g, 8.1mmol) was added, and then TMSOTf (2.2g, 10mmol) was added dropwise, and the reaction was completed and the temperature was raised to room temperature for 4 hours.

To the reaction solution, 130 ml of saturated sodium bicarbonate solution and 100 ml of dichloromethane were added. The layers were separated. The aqueous phase was extracted with dichloromethane (50 ml*2 times). The organic phases were combined, dried and purified by column chromatography to obtain compound 2.24 g. With a yield of 46.3%. ESI(+) m/z: 918.3 [M+Na] ⁺ .

Step 3: Preparation of compound 9

The product of the above step two (2.0g, 2.2mmol) was dissolved in 20ml of methanol, 10ml of lithium hydroxide aqueous solution (1M) was added, and stirred at room temperature overnight.

The reaction solution was neutralized to neutrality and concentrated to dryness under reduced pressure. To the residue were added 30 ml of water and 10 ml of methyl tert-butyl ether. After dissolution, the solution was separated and the aqueous product was prepared by reverse phase C18 to obtain 0.51 g of solid. This is the title compound (Compound 9) with a yield of 38.0%. ESI (+) m/z: 602.2 [M+H] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 8.60-8.55 (2H, Ar-H), 8.25 (1H, s, Ar-H), 5.05 (1H, d), 4.86(1H,d), 4.83-4.76(3H,m), 4.50-4.32(2H,m), 4.08(1H,d), 3.92(1H,t), 3.71-3.69(4H,m), 3.60- 3.50(3H,m), 3.50-3.25(8H,m).

Preparation Example 10 6-mercapto-9-[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]-9H-purine (Compound 10)

Preparation Example 10 was carried out in a similar manner to Preparation Example 9, except that 6-mercaptopurine was substituted for azathioprine to obtain 1.1 g of the title compound. ESI(+) m/z: 477.2 [M+H] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 8.38 (1H, s, Ar-H), 8.18 (1H, s, Ar-H), 5.04 (1H, d), 4.85(1H,d), 4.85-4.75(3H,m), 4.50-4.30(2H,m), 4.08(1H,d), 3.90(1H,t), 3.72(1H,m), 3.60-3.50( 3H, m), 3.50-3.25 (8H, m).

Preparation Example 11 5-fluoro-1-[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]pyrimidine-2,4(1H,3H)-dione (Compound 11)

Preparation Example 11 was carried out in a similar manner to Preparation Example 9, except that azathioprine was replaced with 5-fluorouracil to obtain 0.65 g of the title compound. ESI (-) m/z: 453.1 [MH] ^- .

Preparation Example 12 N-{2-[(S)-1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl]-1,3-dioxo-2 ,3-dihydro-1H-isoindol-4-yl}-N'-[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]acetamide (Compound 12)

Preparation Example 12 was carried out in a similar manner to Preparation Example 9.

step one:

Same as Step 1 of Preparation Example 12.

Step two:

To the reaction flask, add 0.67g of Aprost (1.45mmol), BSA (2.18mmol) and 3ml of acetonitrile at room temperature, heat to reflux for 3h, then cool to room temperature, and add 1.0g of octoacetyl lactulose (compound F, the product of step 1 of Example 9) (1.45 mmol), after stirring for 5 minutes, the solution was dissolved, 0.55 g of TMSOTf (2.47 mmol) was slowly added dropwise, and the mixture was stirred at room temperature for 14 hours.

At 0°C, add 10 ml of saturated sodium bicarbonate solution and 20 ml of dichloromethane to the reaction solution, separate the layers, extract the aqueous phase with dichloromethane (10 ml*2 times), combine the organic phases, dry and concentrate to dryness under reduced pressure. The residue was purified by column chromatography with a dichloromethane/methanol system to obtain a solid product which was heptoacetyl lactulose-aplast glycoside, weighing 263 mg, and a yield of 16.7%. ESI(+) m/z: 1101.4 [M+Na] ⁺ .

Step 3: Preparation of compound 12

100mg of the second product in the above step was dissolved in 5ml of methanol, then 5ml of magnesium methoxide methanol solution (7-8%) was added, stirred at room temperature for 5h, the reaction solution was neutralized with saturated ammonium chloride solution, concentrated to dryness, dissolved in 5ml of water, and filtered The insoluble material was removed, and the filtrate was purified by C18 reverse phase preparation (methanol-water elution). The eluent was concentrated to dryness under reduced pressure to obtain 45 mg of solid as the title compound (Compound 9) in 61.8% yield. ESI(+) m/z: 807.3 [M+Na] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 8.0-7.8 (3H, m Ar-H), 7.2-6.9 (3H, m, Ar-H), 5.79 (1H, m), 5.0-4.53 (5H, m), 4.50-4.20 (4H, m), 4.20-3.83 (6H, m), 3.74 (3H, s), 3.67-3.10 (10H, m), 3.0 (3H, m), 2.51 (3H, s), 1.32 (3H, t).

Preparation Example 13 17-hydroxy-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1,4-diene-3,11,20-trione (Compound 17)

Preparation Example 13 was carried out in a similar manner to Preparation Example 1, except that prednisolone was replaced with prednisone to obtain 1.5 g of the title compound. ESI(+) m/z: 705.3 [M+Na] ⁺ .

Preparation Example 14 4-amino-5-chloro-2,3-dihydro- N- [1-(3-methoxypropyl)-4-pyridinyl] -N' -[4-O-(β-D- (Galactopyranosyl)-D-fructofuranosyl]-7-benzofurancarboxamide (Compound 13)

Step 1: Preparation of Compound K

Octacryl lactulose (Compound A, Preparation Example 1) 2.0g was dissolved in 2.5M hydrogen chloride glacial acetic acid solution (10ml), the reaction was stirred at room temperature for 4h, TLC monitored the disappearance of the raw materials, concentrated at 50°C under reduced pressure to evaporate the solvent The residue was slurried with ether and filtered to obtain crude compound (K), weighing 1.8g, which was directly used in the next reaction.

Step 2: Preparation of Compound M

At room temperature, 1.8 g of crude compound K, 0.54 g (1.47 mmol) of procabili and 10 ml of toluene were mixed and stirred, 3 drops of triethylamine were added dropwise, and the reaction was stirred at 70° C. for 8 h. The reaction solution was concentrated to dryness under reduced pressure and purified by silica gel column chromatography (dichloromethane/methanol) to obtain 0.7 g of product (Compound M) in a yield of 33.6%. ESI(+) m/z: 1442.5 [M+Na] ⁺ .

Step 3: Preparation of compound 13

At room temperature, 0.68 g of compound M (0.48 mmol) was dissolved in 10 ml of tetrahydrofuran, 20 mg (0.37 mmol) of sodium methoxide was added, and the reaction was stirred at room temperature for 4 h. After the reaction was completed, the reaction solution was concentrated and evaporated to dryness under reduced pressure at 45°C, dissolved in 5 ml of water, washed with 5 ml of dichloromethane*3, the aqueous phase was adjusted to pH=8 with dilute acetic acid, and then washed with 5 ml of dichloromethane*2. The resulting product aqueous solution was purified by C18 reverse phase preparation (methanol-water elution), and the collected fractions were concentrated and evaporated to dryness under reduced pressure at 50°C to obtain 0.18 g of a white solid as the title compound (Compound 13) in a yield of 54.2%. ESI (+) m/z: 692.3 [M+H] ⁺ . Chemical purity: 95.0% (HPLC method).

HPLC method:

Chromatography column: octadecylsilane bonded silica gel column; column temperature: 35℃; detection wavelength: 240nm

Mobile phase: A MeOH: CH ₃ CN (2:1)

B 10mmol/L dipotassium hydrogen phosphate (phosphoric acid to adjust pH to 7.1)

Gradient program: 0→20min (B: 70%→45%); 20→25min (B: 45%→70%); 25→29min (B: 70%→70%)

Retention time: 24.1min

Preparation Example 15 5-chloro-2,3-dihydro-4-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]amino}- N -[1-(3-methoxy Propyl)-4-pyridinyl]-7-benzofurancarboxamide (Compound 14)

Step 1: Preparation of Compound N

At room temperature, 20.0 g of octanilide lactulose (16.3 mmol, compound A, Preparation Example 1), 5.0 g of procabulide (13.6 mmol), 40 g of anhydrous sodium sulfate, and 100 ml of acetonitrile were sequentially added to the reaction flask. Stir for 1h under the protection of argon, add tert-butyldimethylsilyl trifluoromethanesulfonate (TBSOTf, 7.1g, 27.2mmol) dropwise, and complete the reaction at room temperature for 3h. Dilute 4.1g of trimethylpyridine with 10ml of dichloromethane and slowly add dropwise to the reaction system to quench the reaction, stir for 30min, filter, wash with dichloromethane, concentrate the filtrate at 45°C under reduced pressure and evaporate to dryness, silica gel column chromatography Purification (dichloromethane/methanol) gave 7.5 g of product (Compound N) in 38.8% yield. ESI(+) m/z: 1442.5 [M+Na] ⁺ .

Step 2: Preparation of compound 14

The product from the above step (Compound N, 1.8 g, 1.27 mmol) was dissolved in 15 ml of tetrahydrofuran at room temperature, 30 mg (0.56 mmol) of sodium methoxide was added, and the reaction was stirred at room temperature for 4 h. After the reaction was completed, the reaction solution was concentrated and evaporated to dryness under reduced pressure at 45°C, dissolved in 5 ml of water, washed with 5 ml of dichloromethane*3, the aqueous phase was adjusted to pH=8 with dilute acetic acid, and then washed with 5 ml of dichloromethane*2. The resulting product aqueous solution was purified by C18 reverse phase preparation (methanol-water elution), and the collected fractions were concentrated and evaporated to dryness under reduced pressure at 50°C to obtain 0.36 g of white solid as the title compound (Compound 14) in a yield of 40.9%. Chemical purity 96.0% (HPLC method). ESI (+) m/z: 692.3 [M+H] ⁺ .

¹ H-NMR (DMSO-d6,400M, BRUKER AV-400): δ (ppm) 7.57-7.45 (2H, m), 5.66 (1H, s), 5.41 (1H, s), 4.75-4.60 (4H, m), 4.34 (1H, m), 4.08 (1H, m), 4.0-3.82 (3H, m), 3.80-3.70 (2H, m), 3.70-3.62 (2H, m), 3.62-3.30 (12H, m), 3.24 (2H, m), 3.21 (3H, s), 2.70 (2H, m), 2.30 (2H, m), 2.04 (2H, m), 1.79 (2H, m), 1.64 (2H, m ), 1.45 (2H, m).

The HPLC method is the same as that in Preparation Example 14, retention time: 13.9min.

Preparation Example 16 4-{ N -methyl- N '-[(3R,4R)-4-methyl-1-(2-cyanoethanoyl)pyridin-3-yl]amino}7-[4-O- (β-D-galactopyranosyl)-D-fructofuranosyl]-7 H -pyrrolo[2,3- d ]pyrimidine (Compound 18)

Preparation Example 16 was carried out in a similar manner to Preparation Example 9, except that azathioprine was replaced with tofacitinib to obtain 0.25 g of the title compound. ESI(+)m/z: 637.5[M+H] ⁺ .

In the following, various formulations (for example, tablets, enteric granules, capsules, etc.) are formulated using the compound represented by formula (I) according to the present invention. For the formulation and specific preparation method, see the following examples.

Formulation Example 1

Using Compound 1 prepared according to the present invention as a pharmaceutical active ingredient, tablets were prepared according to the formulations shown in Table 2 below.

Among them, in the preparation of tablet formulations 1, 3, and 4, all the components of the tablet core are mixed and then compressed.

In the preparation of tablet formulation 2, in addition to magnesium stearate and microcrystalline cellulose, an aqueous solution (5%) of hydroxypropylcellulose is added, wet granulation, drying, and finally adding magnesium stearate for total mixing and tabletting .

In the preparation of tablet formulation 3, the coating was coated with an aqueous dispersion of Opadry (a gastric-coated film coating powder, Shanghai Kale Kang Company). A conventional high-efficiency coating pan is sufficient.

In the preparation of the tablet formulation 4, regarding the preparation of the coating, 20 g of Utech L30D-55 (Evonik_Rhom Rom Resin), 1.8 g of talc, and 0.5 g of triethyl citrate are added in appropriate amounts Dilute with water, stir evenly, and prepare according to the conventional coating process parameters in the efficient coating pot.

Formulation Example 2

Using Compound 1 prepared according to the present invention as a pharmaceutical active ingredient, enteric-coated granules were prepared according to the formulation shown in Table 3 below.

Take compound 1, hypromellose, and talc in the drug formulation to dissolve and disperse in water as a drug solution; place the pellet core in a fluidized bed (WBF-1G Chongqing Yingge Granulation Coating Technology Co., Ltd.) Medium fluidization, setting parameters (air inlet 50m ³ /h, temperature 40~50℃, atomizing pressure 1.0bar), spraying medicine solution at the bottom, flow rate 2~3g/min, controlling material temperature 30±3℃. After the application of the drug, the enteric layer is coated.

The formula of the enteric layer is uniformly mixed and coated according to the above technological parameters to obtain enteric-coated particles.

Formulation Example 3

Using Compound 7 prepared according to the present invention as a pharmaceutical active ingredient, capsules were prepared according to the formulations shown in Table 4 below.

In the preparation of the medicine, after the above contents are mixed evenly, the capsule shell is filled to obtain a capsule.

Formulation Example 4

Using Compound 12 prepared according to the present invention as a pharmaceutical active ingredient, tablets were prepared according to the formulations shown in Table 5 below.

In the process of drug preparation, tablets are all wet granulated. Among them, the tablet core components are mixed first, and then the aqueous solution of the binder is added to wet granulation, after drying, the disintegrant and lubricant are added, the total mixing, and finally the tablet.

The coating adopts the conventional coating process.

Compound performance testing

In the present application, the compounds represented by formula (I) according to the present invention were tested in terms of in vitro stability and systemic pharmacokinetics and local pharmacokinetic characteristics of the gastrointestinal tract in animals.

Stability test of in vitro media

At 37°C, compound 17 was incubated in different media (buffer solutions with different pH, simulated intestinal fluid containing pancreatin and simulated gastric fluid containing pepsin). Samples were taken at intervals as shown in Table 6 below, and the percentage of compound 17 and its degradation products (ie, original form, prednisone) was detected by HPLC method, in which the content of each component was calculated by the normalization method. In the analysis results, the higher the ratio of compound 17 and the lower the ratio of degradation products, the more stable compound 17 is. The results in Table 6 indicate that at a pH above 2.0, within 1.5 hours, the residual percentage of compound 17 is greater than 80%, the degradation product is less than 20%, and has a greater ability to tolerate gastric acid.

Stability in the presence of colonic flora

A phosphate buffer solution with a pH of 7.0 was prepared, and CO ₂ was passed into it to adjust its pH value to 6.8. Then, the cecum contents of male SD rats (weight 180-220g) were taken, added to a pH 6.8 phosphate buffer solution prepared above under N ₂ protection in a water bath at 37°C, so that the concentration was 20% (w /v). Maintain the buffer in an anaerobic environment, then add test compound 17 and stir it evenly.

As shown in Table 7 below, periodically take 0.5 ml of sample solution, add 1.0 ml of acetone to precipitate protein, mix well, let stand for 2 min, and then centrifuge (12000 rpm, 5 min). The supernatant was taken for HPLC analysis, and the content of each component was calculated by the normalization method. As shown in Table 7, Compound 17 was easily degraded in phosphate buffer at pH 6.8, with only 11.3% remaining at 3 hours, which was about 90% degraded. The above results indicate that compound 17 can rapidly degrade and remove glycosidic bonds under the environment of colonic flora to produce an active drug and exert its efficacy in time.

The stability of other typical example compounds was tested according to the above method, and the test results are shown in Table 8 below.

Systemic pharmacokinetics and local pharmacokinetic characteristics of gastrointestinal tract after oral administration of Compound 1 in SD rats

Systemic pharmacokinetics and gastrointestinal tract localization after administration of compound 1 (Preparation Example 1), enteric-coated granules of Compound 1 (Formulation Example 2) and its active ingredient (prednisolone) to SD rats by intragastric administration A pharmacokinetic comparison study to further evaluate the absorption, conversion process, and conversion and release of active ingredients of compound 1 in rats, and to explore whether the application of enteric preparation technology can overcome the pH of compound 1 shown in in vitro tests. The defect of partial degradation in the lower acid environment

According to the experiment, 112 healthy SD male rats (weight 180-220 grams) were selected and divided into 4 groups (prednisolone group, compound 1 group, compound 1 enteric-coated granule group and blank control group), of which prednisone Songlong group, compound 1 group and compound 1 enteric-coated granule group had 36 animals in each group, and 4 animals in blank control group. See Table 9 below for detailed dosing schedule.

For the experimental animals in Group 1, Group 2 and Group 3, at the time points 30min, 1h, 2h, 3h, 4h, 5h, 6h, 8h and 10h after administration, 4 animals were randomly selected at each time point The blood samples were taken and the stomach and various intestines (the first half of the small intestine, the second half of the small intestine, the first 1/3 of the colon (including the cecum) and the second 2/3 of the colon) were taken and weighed. The concentration of prednisolone and compound 1 in plasma and contents was determined by LC-MS/MS method, and the content of prednisolone and compound 1 in each section of the digestive tract was calculated according to the weight of the contents. Four animals in group 4 were sacrificed at the 10-hour time point, and blood and the contents of the gastrointestinal tract were used for blank control.

The distribution of prednisolone in various parts of the gastrointestinal tract of rats at different time points after administration in the prednisolone group is shown in Table 10 below. From the results in Table 10, it can be seen that after prednisolone is given by gavage in rats, the prednisolone in the gastrointestinal tract is mainly distributed in the stomach and small intestine, with the highest concentration in the stomach; and the contents of the cecum and colon The concentration of nisone is very low.

The distribution of prednisolone in the contents of various parts of the gastrointestinal tract of rats in the compound 1 and compound 1 enteric-coated granule groups after intragastric administration is shown in Table 11 and Table 12, respectively. Compared with the prednisolone group, the gastrointestinal distribution of the compound 1 group and the compound 1 enteric-coated granule group was significantly reduced, while the local maximum concentration of prednisolone (Cmax) in the cecum and colon increased by 65-360 times (result (See Table 13), the total exposure (AUC) is also much higher than the prednisolone prototype drug group (see Figure 1 and Figure 2 for drug-time curves). Enteral-coated granules were used to avoid the partial release of compound 1 in the acidic environment of the stomach, and the compound 1 enteric-coated granule group basically did not release prednisolone in the stomach and anterior part of the small intestine. Detected), while the local prednisolone concentration in the colon was slightly higher than the compound 1 group. The above results indicate that the main release site of oral compound 1 enteric-coated granules in the body is the cecum and colon, followed by the posterior small intestine site close to the cecum, which is consistent with the purpose of the compound of the present invention.

Figure 3 shows the change curve of plasma prednisolone concentration in the prednisolone group, compound 1 group and compound 1 enteric-coated granule group after intragastric administration. From this drug-time curve, it can be seen that the Cmax value of the compound 1 group and the compound 1 enteric-coated granule group is very low compared with the free prednisolone group, and the time to reach the peak is also significantly delayed.

Compound 1 rats had a lower concentration of compound 1 (10.4 ng/ml) at a time point of only 0.5 hours in the plasma. After administration of compound 1 enteric-coated particles, compound 1 was not detected at all time points. The results of this study indicate that Compound 1 is only slightly absorbed in the stomach or upper small intestine of rats. The use of enteric-coated preparation technology to avoid gastric release can effectively avoid the absorption of Compound 1.

Pharmacodynamic effect of compound 1 by intragastric administration on trinitrobenzenesulfonic acid (TNBS) enema-induced Wistar rat ulcerative colitis model

By conducting intragastric administration of Compound 1 and the prednisolone marketed preparation (prednisolone tablets, produced by Wuhan Yuancheng Co-Creation Technology Co., Ltd.), a comparative study on the therapeutic effect of Wistar rats with ulcerative colitis was further studied. Evaluate the improvement of the main therapeutic index of compound 1 intragastric therapy on ulcerative colitis and make a systematic comparison with the treatment group of prednisolone, etc., to provide pharmacodynamics for the further development of compound 1 support.

According to this experiment, 40 male Wistar rats of 180-220 grams were selected and randomly divided into four groups (saline control group, model control group, prednisolone group and compound 1 group), 10 animals in each group. Except for the physiological saline control group, the other three groups of experimental animals used the method of one-time enema of 3.2ml/kg 2.5% TNBS-50% ethanol mixed solution (containing 80mg/kg TNBS) to make animal models of ulcerative colitis. The normal saline control group had a one-time enema of 3.2ml/kg normal saline. Twenty-four hours after the administration of the model, the corresponding vehicle control or therapeutic drugs (qd×7d (administer once a day, continuous administration for 7 days)) are administered by gavage in groups according to the animals. For detailed grouping and administration information, please refer to Table 14 below .

The main observation indicators during the study include: animal death, clinical observation of the animal, body weight, colon coefficient, histopathological examination, etc. Each experimental group was sacrificed on the 8th day after modeling, and the colon tissue was taken to calculate the colon coefficient (colon coefficient = colon weight (g)/animal weight (g)*100%), and then histopathological examination and scoring (tissue Pathology scoring criteria are shown in Table 15 below).

During the course of the study, only one animal in the model control group died on the 6th day after the model was made, and no abnormal animal death was found in the other groups.

The clinical symptoms of model animals are mainly: perianal orange/yellow liquid, blood stains around the nasal cavity, blood stains at the corners of the eyes, bow hairs, loose stools, lack of energy, reduced activity, slow movements, yellow/orange loose stools , Brown loose stools, bloody loose stools and no stool etc.

As shown in Figure 4, throughout the test period, the body weight of the negative control group continued to increase; on the contrary, the model control, prednisolone, and compound 1 groups had an average of animals in the first 3 days after modeling. Body weight showed a significant downward trend, and the weight of compound 1 group began to rebound from the 4th day of modeling and the other two groups began from the 5th day of modeling. The body weight of compound 1 group recovered faster than the other two groups. Further, a one-way analysis of variance was used to statistically compare the weight change on the 8th day (weight change = weight on the 8th day (g)-weight on the 1st day (g)) of different experimental groups (see Figure 5, where * indicates p<0.05, compared with the negative control group; # means p<0.05, compared with the model control group), the results show that the weight gain of the three groups of animals modeled with TNBS are significantly lower than the negative control group with saline model; and the model Compared with the group, the body weight gain of the prednisolone group and the compound 1 group was higher than that of the model group, but only the difference between the compound 1 group and the model group was statistically significant. This result shows that the same mole number of compound 1 has a greater effect on TNBS The enema-induced ulcerative colitis rat model has better therapeutic effect than the prednisolone prototype.

The index of the colon coefficient was obtained after dissecting the animal on the 8th day after the model was constructed. The colon coefficient of each experimental group was compared using a one-way analysis of variance. The results are consistent with the analysis results of the change in body weight (see Figure 6, where * indicates p<0.05, compared with the negative control group; # means p<0.05, compared with the model control group). Compared with the negative control group, the colon coefficient of each model group was significantly increased (p<0.05), the model control group was the most obvious, the prednisolone group was the second, and the compound 1 group had the lowest increase; further prednisolone Compared with the compound 1 group and the model control group, the colon coefficient of the compound 1 group was significantly lower than that of the model control group (p<0.05), while the difference between the prednisolone group and the model control group did not reach statistical significance.

On the 8th day after TNBS modeling, the histopathological changes of the colon were mainly manifested as focal mucosal necrosis in the descending colon, infiltration of neutrophils in the mucosa and submucosa, edema, vasodilation, and hyperemia.

The results of single-factor analysis of variance of colon histopathology scores in each experimental animal group are shown in Figure 7 (where * indicates p<0.05, compared with the negative control group; # indicates p<0.05, compared with the model control group), as compared with the negative control Compared with the group, the histopathological scores of each model group were significantly increased (p<0.05), the model control group was the most obvious, the prednisolone group was the second, and the compound 1 group had the lowest increase; further prednisolone and Compared with the model control group, the histopathology of the compound 1 group was significantly lower than that of the model control group (p<0.05), while the difference between the prednisolone group and the model control group did not reach statistical significance. This result is consistent with the aforementioned analysis results of body weight and colon coefficient, further suggesting that the administration of an equal molar amount of Compound 1 to the rat model of ulcerative colitis is superior to prednisolone.

The above studies have been conducted on compound 1 in terms of systemic pharmacokinetics, local pharmacokinetics, and pharmacodynamics of the animal body system, and the results prove that the lactulose glycoside compound 1 according to the present invention can pass through the mammalian gastrointestinal tract It is not significantly absorbed by the gastrointestinal tract and is not significantly hydrolyzed by endogenous enzymes of the mammalian host, so the lactulose glycoside compound 1 can reach the mammalian colon and release the active drug in the colon under the action of the colonic flora. The lactulose glycoside compound 1 has a colon-specific drug release effect and can be used for the prevention or treatment of intestinal diseases.

It should be noted that the present invention also carries out the rest of the compounds (ie, compounds 2-17 shown in Table 1) in animal pharmacokinetics, local pharmacokinetics of the gastrointestinal tract, and pharmacodynamic effects, etc. Experimental research. The results prove that the lactulose glycoside compound 2-17 according to the present invention can also pass through the gastrointestinal tract of mammals without being significantly absorbed by the gastrointestinal tract and is not significantly hydrolyzed by endogenous enzymes of the mammalian host, so the lactulose glycoside compound 2-17 It can reach the colon of mammals and release active drugs in the colon under the action of colon flora. The lactulose glycoside compounds 2-17 have the effect of localized drug release in the colon, and can be used for the prevention or treatment of intestinal diseases.

Although the present invention has been described in detail above with reference to examples and comparative examples, it should be noted that the present invention is not limited to the examples and comparative examples, and that the present invention can be changed or modified without departing from the spirit of the present invention.

Claims (15)

A compound represented by formula (I),

Among them, the lactulose group is connected to the heteroatom in the aglycon (G) through a glycosidic bond, which is a group formed by removing one hydrogen atom from the heteroatom of the active drug molecule, and "

"Indicates that the lactulose group is connected to the heteroatom in the aglycon (G) through an alpha glycosidic bond or a beta glycosidic bond. The compound as described in item 1 of the patent application scope, wherein the active drug molecule is selected from the group consisting of glucocorticoid drugs, Apostat, procapril, mesalazine, metronidazole, azathioprine, Any one of 6-mercaptopurine, 5-fluorouracil, and tofacitinib. The compound according to item 1 of the patent application scope, wherein the heteroatom system is selected from any one of an oxygen atom, a nitrogen atom, and a sulfur atom. The compound as described in item 1 of the patent application, wherein the aglycon (G) is obtained by removing one hydrogen atom from the alcohol hydroxyl group, phenolic hydroxyl group, amino group, amide group or NH on the heterocyclic ring in the active drug molecule The formed group. The compound according to item 2 of the patent application scope, wherein the glucocorticoid drugs are selected from cortisone acetate, prednisone, prednisone acetate, hydrocortisone, hydrocortisone acetate, butyric acid Hydrocortisone, hydrocortisone sodium succinate, hydrocortisone sodium phosphate, fludrocortisone, fludrocortisone acetate, prednisolone, prednisolone acetate, prednisolone succinic acid Sodium, prednisolone sodium phosphate, betamethasone, betamethasone acetate, betamethasone dipropionate, clobetasol butyrate, betamethasone valerate, betamethasone sodium phosphate, beclomethasone dipropionate , Mometasone furoate, clobetasol propionate, dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, p-flumethasone, fluticasone propionate, methylprednisolone, methylprednisolone acetate, flumidon, Qu Any of Ansiron, Triamcinolone acetonide, Triamcinolone acetonide, Flucinide, Ancinide, Hasinaide, Ciclesonide, Budesonide, and Defecote. The compound according to item 2 of the patent application scope, wherein the glucocorticoid drugs are selected from hydrocortisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone and budeson Any of Ned. The compound as described in item 1 of the patent application, wherein the aglycon (G) is a group represented by the following formula (II):

Where R ₁ is H or F; R ₂ is H, F or -CH ₃ ; R ₃ is -OH; and R ₄ is H, -OH or -CH ₃ ; or R ₃ and R _{4 are} connected to form a group

,

or

. The compound according to item 7 of the patent application scope, wherein the aglycone (G) is selected from any one of the following:

Where Me represents -CH ₃ . The compound as described in item 1 of the patent application scope, wherein the aglycone (G) is selected from any one of the following:

Where Me represents -CH ₃ . The compound as described in item 1 of the patent application scope, wherein the aglycone (G) is selected from any one of the following:

Where Me represents -CH ₃ . The compound as described in Item 1 of the patent application scope, wherein the compound is any one selected from the following: (11β)-11,17-dihydroxy-21-{[4-O-(β-D -Galactopyranosyl)-D-fructofuranosyl]oxy}progesterone-1,4-diene-3,20-dione; (11β)-11,17-dihydroxy-21-{[4- O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-4-ene-3,20-dione; (6α,11β)-11,17-dihydroxy-6 -Methyl-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1,4-diene-3,20-dione; ( 11β,16α)-9-fluoro-11,17-dihydroxy-16-methyl-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnancy Steroid-1,4-diene-3,20-dione; (11β,16β)-9-fluoro-11,17-dihydroxy-16-methyl-21-{[4-O-(β-D -Galactopyranosyl)-D-fructofuranosyl]oxy}progesterone-1,4-diene-3,20-dione; (11β,16α)-9-fluoro-11,16,17-tri Hydroxy-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1,4-diene-3,20-dione; (11β, 16α)-11-hydroxy-16,17-(butylene dioxy)-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl Ioxy}pregnant-1 ,4-diene-3,20-dione; 2-methyl-5-nitro-1-{2-{[4-O-(β-D-galactopyranosyl)-D-fructose Yl]oxy}ethyl}imidazole; 6-[(1-methyl-4-nitro- 1H -imidazol-5-yl)sulfur]-9-[4-O-(β-D-pyran half Lactosyl)-D-fructofuranosyl]-9 H -purine; 6-mercapto-9-[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]-9 H -purine ; 5-fluoro-1-[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]pyrimidine-2,4(1H,3H)-dione; N -{2-[ (S)-1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl]-1,3-dioxo-2,3-dihydro- 1 H -isoindol-4-yl}- N' -[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]acetamide; 4-amino-5-chloro- 2,3-dihydro- N- [1-(3-methoxypropyl)-4-pyridinyl] -N' -[4-O-(β-D-galactopyranosyl)-D -Fructofuranosyl]-7-benzofurancarboxamide; 5-chloro-2,3-dihydro-4-{[4-O-(β-D-galactopyranosyl)-D-frufuranose Radical]amino}- N -[1-(3-methoxypropyl)-4-pyridinyl]-7-benzofurancarboxamide; 5-amino-2-{[4-O-(β-D-pyran half Lactosyl)-D-fructofuranosyl]oxy}benzoic acid; 2-hydroxy-5-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]amino}benzoic acid; 17-hydroxy-21-{[4-O-(β-D-galactopyranosyl)-D-fructofuranosyl]oxy}pregnant-1,4-diene-3,11,20-trione ; And 4-{ N -methyl- N '-[(3R,4R)-4-methyl-1-(2-cyanoacetoxy)pyridin-3-yl]amino}7-[4- O-(β-D-galactopyranosyl)-D-fructofuranosyl]-7 H -pyrrolo[2,3- d ]pyrimidine. A pharmaceutical composition comprising the compound as described in any one of claims 1 to 11 and a pharmaceutically acceptable carrier. A use of the compound as described in any one of patent application items 1 to 11 for the preparation of a medicament for preventing or treating intestinal diseases. The use as described in item 13 of the patent application scope, wherein the intestinal disease is selected from ulcerative colitis, Crohn's disease, infectious enteritis, irritable bowel syndrome, chronic constipation, amoebic bowel disease, Colon cancer and rectal cancer. The use according to item 13 of the patent application scope, wherein the intestinal disease is selected from ulcerative colitis, Crohn's disease and chronic constipation.

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