阅读说明：本技术 一种具有抗菌活性的胆酸衍生物及其药用组合物 (Cholic acid derivative with antibacterial activity and medicinal composition thereof ) 是由 广兵 阳泰 董韧涵 刘进 覃传军 黄胜 谢建 赖永新 彭向阳 许庆 占伟 彭坚 于 2019-11-07 设计创作，主要内容包括：本发明提供了式(I)所示的化合物、或其立体异构体、或其盐、或其溶剂合物、或其前体药物、或其代谢产物。该类化合物能够有效抑制艰难梭菌菌体的生长,具有显著的抑菌活性,在制备预防和/或治疗艰难梭菌感染性疾病、艰难梭菌感染性疾病的复发、或艰难梭菌感染性疾病并发症的药物中具有非常好的应用前景。<Image he="391" wi="700" file="DDA0002264882660000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The present invention provides a compound represented by formula (I), or a stereoisomer thereof, or a salt thereof, or a solvate thereof, or a prodrug thereof, or a metabolite thereof. The compound can effectively inhibit the growth of the thalli of clostridium difficile, has obvious bacteriostatic activity, and has very good application prospect in preparing medicaments for preventing and/or treating clostridium difficile infectious diseases, relapse of clostridium difficile infectious diseases or complications of clostridium difficile infectious diseases.)

1. A compound represented by the formula (I), or a stereoisomer thereof, or a salt thereof, or a solvate thereof, or a prodrug thereof, or a metabolite thereof:

wherein m is 0 or 1, n is 0, 1 or 2;

R₇is selected from-NO₂Halogen, -OH, -NH₂、C_1-5Alkyl of (C)_1-5Alkoxy group of (a);

R₈selected from H, -NO₂Halogen, -OH, -NH₂、C_1-5Alkyl of (C)_1-5Alkoxy group of (a);

y is nothing, or Y is selected from the structures shown as the following Y-1 or Y-2:

wherein R is₆Is selected from-H, C_1-4Alkyl, -OH, -OCH₃、-COOCH₃；

R₉Selected from H, C_1-3Alkyl, benzyl, C_1-3Hydroxyalkyl of (C)_1-3Mercaptoalkyl groups of (a).

2. The compound according to claim 1, or a stereoisomer thereof, or a salt thereof, or a solvate thereof, or a prodrug thereof, or a metabolite thereof, wherein: the structure of the compound is shown as the formula (II):

wherein n is 0, 1, 2;

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl。

3. A compound according to claim 2, or a stereoisomer thereof, or a salt thereof, or a solvate thereof, or a prodrug thereof, or a metabolite thereof, wherein: the structure of the compound is shown as a formula (II-a):

wherein R is₅Is hydrogen or hydroxy;

n＝0、1、2；

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl。

4. The compound according to claim 1, or a stereoisomer thereof, or a salt thereof, or a solvate thereof, or a prodrug thereof, or a metabolite thereof, wherein: the structure of the compound is shown as the formula (III):

wherein n is,

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl；

R₉Is selected from-H, C_1-3Alkyl, benzyl, C substituted by hydroxy or mercapto_1-3Alkyl groups of (a);

preferably, the compound has the structure of formula (IIIa), formula (IIIb) or formula (IIIc):

wherein n is,

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl。

5. The compound according to claim 1, or a stereoisomer thereof, or a salt thereof, or a solvate thereof, or a prodrug thereof, or a metabolite thereof, wherein: the structure of the compound is shown as the formula (IV):

preferably, the structure of the compound is represented by formula (IVa) or formula (IVb):

wherein n is,

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl。

6. The compound according to any one of claims 1 to 5, or a stereoisomer thereof, or a salt thereof, or a solvate thereof, or a prodrug thereof, or a metabolite thereof, wherein: selected from the following structures:

7. use of a compound according to any one of claims 1 to 6 in the preparation of a bacteriostatic agent.

8. Use according to claim 7, characterized in that: the bacteriostatic agent is a medicament for inhibiting the thalli of clostridium difficile.

9. Use according to claim 8, characterized in that: the medicament can prevent and/or treat clostridium difficile infectious diseases, relapse of clostridium difficile infectious diseases, or complications of clostridium difficile infectious diseases.

10. Use according to claim 9, characterized in that: the clostridium difficile infectious disease, the recurrence of clostridium difficile infectious disease, or the complication of clostridium difficile infectious disease is caused by infection of clostridium difficile thallus; the clostridium difficile infectious disease complication is a digestive tract infection syndrome caused by clostridium difficile thallus infection.

11. Use according to claim 10, characterized in that: the digestive tract infection syndrome includes, but is not limited to, pseudomembranous enteritis, diverticulitis, antibiotic-associated diarrhea, incomplete or complete ileus.

12. A drug for inhibiting Clostridium difficile bacterial cells, which is characterized in that: the medicament is a preparation prepared by taking the compound of any one of claims 1-6 as an active ingredient and adding pharmaceutically acceptable auxiliary materials.

13. The pharmaceutical composition of claim 12, wherein: the pharmaceutically acceptable auxiliary materials are selected from any one or more than two of diluents, fillers, coloring agents, glidants, lubricants, binders, stabilizers, suspending agents or buffering agents.

14. The medicament according to claim 12 or 13, characterized in that: the preparation is an oral preparation; preferably, the oral formulation is selected from granules, capsules, tablets, pills.

15. The medicament according to any one of claims 12 to 14, characterized in that: the dosage of the active ingredient contained in the pharmaceutical unit preparation is 5-2500 mg.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a cholestane derivative with antibacterial activity and a medicinal composition thereof.

Background

Clostridium difficile (Clostridium difficile) is an obligate anaerobe of the genus Clostridium, is very sensitive to oxygen and is difficult to separate and culture, so it is named as being generally parasitic in human intestinal tract. Difficile infection is usually caused by overuse of certain antibiotics, which breaks the balance of intestinal flora, increases the growth rate of the flora of difficile and causes inflammation. Difficile produces exotoxins a and B and produces different effects at different times. Toxin a is enterotoxin, which binds to mucosal cells at first in the early stage, causing primary destruction, causing inflammation of the intestinal wall, cell infiltration, increased intestinal wall permeability, bleeding and necrosis. Toxin B is a cytotoxin, damages cytoskeleton, causes cell shrinkage necrosis, directly damages intestinal wall cells and causes diarrhea.

Difficile infectious disease is a disease caused by infection with the bacteria and/or spores of clostridium difficile. Pseudomembranous enteritis is a common clostridium difficile infectious disease, and is clinically manifested by diarrhea, abdominal pain and systemic poisoning symptoms. Symptoms begin suddenly and are accompanied by hypotension, usually with fever, leukocytosis and even death. In addition, infection with the bacteria and/or spores of clostridium difficile can cause complications of clostridium difficile infectious diseases, common complications including pyelonephritis, meningitis, abdominal and vaginal infections, bacteremia, and gas gangrene. In recent years, clostridium difficile has become an important pathogenic bacterium causing nosocomial infectious diseases, and is increasingly emphasized.

The current main means for treating clostridium difficile infectious diseases is to use bacteriostatic agents. Metronidazole and vancomycin are the two most commonly used bacteriostatic agents, but patients with clostridium difficile infectious diseases treated by metronidazole and vancomycin have the possibility of relapse, and the prognosis is very poor after relapse. More importantly, given that bacterial resistance under evolutionary pressure can in time constitute a major therapeutic challenge, and that the emergence of metronidazole-and vancomycin-resistant refractory strains has been observed clinically, the development of bacteriostatic agents with high activity that inhibit new mechanisms or structures of clostridium difficile is elusive.

The cholic acid compounds are tetracyclic condensed ring steroid compounds with similar structures and generally containing 24 carbon atoms, such as decarbonized (nor) cholic acid with 1 carbon atom degraded by side chain so that the total carbon number is 23, homotypic (homo) cholic acid with 25 carbon atoms increased by one carbon atom, and the cholic acid compounds or derivatives chemically modified on the basis of the structures of the cholic acid compounds have more biological activities, and ursodeoxycholic acid is an endogenous cholic acid compound which is approved to treat cholestatic cirrhosis at present, and other cholic acid derivatives have activities of inhibiting germination of clostridium difficile spores, such as a compound CamSA (named as 3- [ (3, 7 α,12 α -trihydroxy-24-oxo-5 β -cholan-24-yl) amino ] benzazenen acid) reported in published in journal of Infects 2013, 207:1498-504 and a compound named as Medxy-24-oxo-5-cholan-24-yl) amide ] chezon acid (named as 3, 7, α, 12-trihydroxy-6712-3, 6712-3.

Disclosure of Invention

The invention aims to provide a novel cholic acid compound which has high activity and can inhibit clostridium difficile thallus and a medicinal composition thereof.

The present invention provides a compound represented by formula (I), or a stereoisomer thereof, or a salt thereof, or a solvate thereof, or a prodrug thereof, or a metabolite thereof:

wherein m is 0 or 1, n is 0, 1 or 2;

represents a single bond or a double bond,

each independently selected from-H, -OH, -NH₂、＝O、＝NOH、C_1-5Alkyl groups of (a);

R₇is selected from-NO₂Halogen, -OH, -NH₂、C_1-5Alkyl of (C)_1-5Alkoxy group of (a);

R₈selected from H, -NO₂Halogen, -OH, -NH₂、C_1-5Alkyl of (C)_1-5Alkoxy group of (a);

y is nothing, or Y is selected from the structures shown as the following Y-1 or Y-2:

wherein R is₆Is selected from-H, C_1-4Alkyl, -OH, -OCH₃、-COOCH₃；

R₉Selected from H, C_1-3Alkyl, benzyl, C_1-3Hydroxyalkyl of (C)_1-3Mercaptoalkyl groups of (a).

Further, the structure of the compound is shown as the formula (II):

wherein n is 0, 1, 2;

each independently selected from-H, -OH, -NH₂、＝O、＝NOH、C_1-5Alkyl groups of (a);

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl。

Further, the structure of the compound is shown as the formula (II-a):

wherein R is₅Is hydrogen or hydroxy;

n＝0、1、2；

each independently selected from-H, -OH, -NH₂、＝O、＝NOH、 C_1-2Alkyl groups of (a);

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl。

Further, the structure of the compound is shown as the formula (III):

wherein n is,

As described above;

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl；

R₉Is selected from-H, C_1-3Alkyl, benzyl, C substituted by hydroxy or mercapto_1-3Alkyl groups of (a);

preferably, the compound has the structure of formula (IIIa), formula (IIIb) or formula (IIIc):

wherein n is,

As described above;

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl。

Further, the structure of the compound is shown as the formula (IV):

preferably, the structure of the compound is represented by formula (IVa) or formula (IVb):

wherein n is,

R₆As described above;

R₇is selected from-NO₂、Cl；R₈Selected from H, -NO₂、Cl。

Further, the compound is selected from the following structures:

the invention also provides application of the compound in preparing a bacteriostatic agent.

Furthermore, the bacteriostatic agent is a medicament for inhibiting the thalli of clostridium difficile.

Further, the medicament is capable of preventing and/or treating clostridium difficile infectious diseases, recurrence of clostridium difficile infectious diseases, or complications of clostridium difficile infectious diseases.

Further, the clostridium difficile infectious disease, recurrence of clostridium difficile infectious disease, or complication of clostridium difficile infectious disease is caused by infection with clostridium difficile bacteria; the clostridium difficile infectious disease complication is a digestive tract infection syndrome caused by clostridium difficile thallus infection.

Further, the syndromes of digestive tract infections include, but are not limited to, pseudomembranous enteritis, diverticulitis, antibiotic-associated diarrhea, incomplete or complete ileus.

The invention also provides a medicament for inhibiting the thalli of clostridium difficile, which is a preparation prepared by taking the compound as an active ingredient and adding pharmaceutically acceptable auxiliary materials.

Further, the pharmaceutically acceptable auxiliary materials are selected from any one or more than two of diluents, fillers, coloring agents, glidants, lubricants, binders, stabilizers, suspending agents or buffering agents.

Further, the formulation is an oral formulation; preferably, the oral formulation is selected from granules, capsules, tablets, pills.

Further, the active ingredient is contained in the pharmaceutical unit preparation in an amount of 5-2500 mg.

In the present invention, the "salt" is an acid and/or base salt of a compound or a stereoisomer thereof with an inorganic and/or organic acid and/or base, and also includes a zwitterionic salt (inner salt), and also includes a quaternary ammonium salt such as an alkylammonium salt. These salts can be obtained directly in the final isolation and purification of the compounds. The compound, or a stereoisomer thereof, may be obtained by appropriately (e.g., equivalentlymixing) a certain amount of an acid or a base. These salts may form precipitates in the solution which are collected by filtration, or they may be recovered after evaporation of the solvent, or they may be prepared by reaction in an aqueous medium followed by lyophilization. The salt in the invention can be hydrochloride, sulfate, citrate, benzene sulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

Prefix (C)_a-b) Alkyl of (a) indicates any alkyl group containing from "a" to "b" carbon atoms. E.g. C_1-5The alkyl group of (b) is a straight-chain or branched alkyl group having 1 to 5 carbon atoms.

Experiments prove that the compound provided by the invention can effectively inhibit the growth of clostridium difficile thalli, has obvious antibacterial activity, and has a very good application prospect in preparation of medicines for preventing and/or treating clostridium difficile infectious diseases, relapse of clostridium difficile infectious diseases or clostridium difficile infectious disease complications.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Detailed Description

The raw materials and equipment used in the invention are known products, and are obtained by purchasing commercially available products or preparation methods in reference publications.

The cholic acid compound is commercially available (HPLC purity is more than 98%), and comprises CA (cholic acid), CDCA (chenodeoxycholic acid), DCA (deoxycholic acid), LCA (lithocholic acid), UDCA (ursodeoxycholic acid), HCA (hyocholic acid), HDCA (hyodeoxycholic acid), OCA (octocholic acid), PCA (phosphonodeoxycholic acid), and 7-ketoLCA (7-carbonyllithocholic acid), DHCDCA (dehydrodeoxycholic acid), DHCA (dehydrocholic acid), 7-ketoDCA (7-carbonylcholic acid), 3-DHDCA (3-dehydrocholic acid) and 3-DHCA (3-dehydrocholic acid) which correspond to the oxo-carbonyl structure of bile acid of natural origin. The abbreviations for the compounds and the corresponding structures are as follows:

the decarbonized (nor) cholic acid raw material has the following structure and code, and is represented by nor-CA (decarbonized cholic acid), nor-CDCA (decarbonized chenodeoxycholic acid), nor-UDCA (decarbonized ursodeoxycholic acid), nor-7DHCA (decarbonized 7-dehydrocholic acid), 7-keto-norLCA (decarbonized 7-carbonyl lithocholic acid), and the synthesis method is carried out according to the method of Journal of Lipid Research,1988,29: 1387: