CN106928308B - Prodrug of auriculatone and application thereof - Google Patents

Abstract

The invention discloses derivatives of compounds of general formula (I), wherein R1 is amino acid ester, phosphate, sulfate, sulfonate, carboxylate, or pharmaceutically acceptable salts of these esters; r2 is an oxygen atom or a group of formula II, wherein A is a hydroxyl group or a group of formula III; r3 is proton, alkyl, phenyl or aralkyl; r4 is proton, alkyl, phenyl or aralkyl, carboxy or substituted carboxy, phosphate, sulfate, sulfonate or pharmaceutically acceptable salt thereof; wherein R is₁、R₂、R₃、R₄The prodrug derived from the general formula (I) containing the derivative can be used for preventing and treating liver injury.

Description

Prodrug of auriculatone and application thereof

Technical Field

The invention relates to a novel water-soluble medicine for preventing and treating liver injury and suitable for oral administration or injection administration, in particular to a prodrug obtained by derivatization of hydroxyl at 3 position or carbonyl at 16 position of auriculatone.

Background

Auriculatone is well known in the existing research as a medicine for preventing, protecting and treating liver injury. Of the various types known, one particularly effective type is the hydroxyl-containing group at position 3 and the ketocarbonyl-containing group at position 16. For example, the patent numbers of the disclosed Chinese invention patents are: 201210065753.1, the name is: an application of auriculatone in preparing a medicine for preventing and treating liver injury, which discloses that the medicine has the functions of preventing, protecting and treating liver injury with the chemical name of 3-hydroxy-17 alpha-H, 18 beta-H-28-norolean-12-eme-16-one.

However, the application of this type of drug is limited by its low water solubility, which prevents its development into suitable dosage forms for oral gastrointestinal administration, injection, etc.

Disclosure of Invention

In view of the above deficiencies of the prior art, a novel water-soluble drug or a pharmaceutically acceptable salt thereof for preventing and treating liver damage and suitable for oral or injectable administration is provided.

Specifically, the invention provides a compound shown in the following general formula I, a pharmaceutical composition for preventing, protecting and treating liver injury, which comprises a therapeutically effective amount of the compound shown in the general formula I and a pharmaceutically acceptable carrier, and the compound provides more drug selection ways in the aspects of preventing, protecting and treating liver injury.

The novel water-soluble drugs for preventing and treating liver damage according to the present invention and suitable for oral or injectable administration, or pharmaceutically acceptable salts thereof, are represented by the following general formula (I):

wherein R1 is an amino acid ester, phosphate, sulfate, sulfonate, carboxylate, or a pharmaceutically acceptable salt of such an ester; r2 is an oxygen atom or a group of formula II, wherein A is a hydroxyl group or a group of formula III; r3 is proton, alkyl, phenyl or aralkyl; r4 is proton, alkyl, phenyl or aralkyl, carboxyl or substituted carboxyl, phosphate, sulfate, sulfonate or pharmaceutically acceptable salt thereof

Preferably, wherein R1 and R4 are pharmaceutically acceptable salts of sodium, potassium, calcium, ammonium of amino acid esters, phosphate esters, sulfate esters, sulfonate esters, or carboxylate esters.

Preferably, wherein R1 and R4 are amino acid ester moieties of the general formula (IV) wherein amino acids include aliphatic amino acids, aromatic amino acids, heterocyclic amino acids and heterocyclic imino acids and derivatives thereof;

preferably, the aliphatic amino acid comprises alanine, valine, leucine, isoleucine, methionine, aspartic acid, glutamic acid, lysine, arginine, glycine, serine, threonine, cysteine, asparagine, or glutamine; the aromatic amino acid comprises phenylalanine or tyrosine; the heterocyclic amino acid comprises histidine or tryptophan; the heterocyclic imino acid comprises proline.

Preferably, wherein R1 and R4 are partial structures of sulfuric acid esters of the general formula (V) wherein R7 may be a proton, alkyl, phenyl or aralkyl group;

preferably, R1 and R4 are partial structures of phosphates of the general formula (VI) wherein R8, R9 may be protons or alkyl, phenyl, aralkyl;

preferably, R1 and R4 are part structures of a sulfonate of the general formula (VII) wherein R10 is hydrogen, alkyl, phenyl or aralkyl;

preferably, R1 and R4 are partial structures of a carboxylic ester of the following general formula (VIII), wherein R11 is hydrogen, alkyl, phenyl or aralkyl, carboxylic ester containing a carboxyl group;

alkyl as used herein refers to straight or branched chain saturated aliphatic groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl and the like; the phenyl group refers to a group in which one or more hydrogens on the phenyl ring are replaced by a methyl group, an ethyl group, an n-propyl group, an isopropyl group, such as an o-methylphenyl group, a p-methylphenyl group, a m-methylphenyl group, a 2, 3-dimethylphenyl group, etc.; the aralkyl group means a group in which a phenyl group replaces a hydrogen on a straight or branched saturated aliphatic group, such as a benzyl group, a phenethyl group and the like.

The term "pharmaceutically acceptable salts" as used herein includes ammonium phosphates, metal salts, salts with amino acids, salts with amines and salts with other bases such as piperidine or morpholine. The term "pharmaceutically acceptable salts" includes mono-and di-salts. Specific embodiments include ammonium, sodium, calcium, magnesium, cesium, lithium, potassium, barium, zinc, aluminum, lysine, arginine, histidine, methylamine, ethylamine, tert-butylamine, cyclohexylamine, N-methylglucamine, ethylenediamine, glycine, procaine, benzathene, diethanolamine, triethanolamine, piperidine, and morpholine.

Compared with the prior art, the invention has the beneficial technical effects that: 1. the pro-drug shows significantly improved water solubility over the parent compound, making it useful for injection and oral administration, the compound being stable in solution and converted to the parent drug in vivo; 2. the medicaments of the invention may be used alone or formulated in pharmaceutical compositions comprising, in addition to the active ingredient, a pharmaceutically acceptable carrier, adjuvant, slow-release or diluent. The compounds may be administered by various methods, for example by the oral or injectable routes. The pharmaceutical compositions may be in solid form such as capsules, tablets, powders and the like, or in liquid form such as solutions, suspensions or emulsions. Compositions for injection may be presented in unit dosage form in ampoules or in multi-dose containers, and may contain additives such as suspending, stabilizing and dispersing agents. The compositions may be in a ready-to-use form or in powder form for reconstitution with a suitable excipient, e.g. sterile water, at the time of administration; 3. the drug of the present invention has pharmacological activity in animals, including mammals, and particularly humans, and is useful for the prevention, protection, and treatment of liver damage. The medicine greatly increases the solubility of parent medicines for preventing and protecting hepatic injury, and is proved to release the parent compounds with biological activity (namely playing the role of prodrugs) in mouse experiments.

Detailed Description

The technical solutions in the embodiments of the present invention are clearly and completely described below. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. Some abbreviations used in the detailed description of the invention are as follows:

h is h; PEG ═ polyethylene glycol; mg/ml-mg per ml; mg/kg-mg/kg; APAP ═ acetaminophen; NAC-N-acetylcysteine; rpm is equal to revolutions per minute; min is minutes; ALT ═ glutamic-pyruvic transaminase; AST ═ glutamic-oxalacetic transaminase; IU/L is international units per liter; g is gram; degree centigrade. In the following examples, all temperatures are in degrees Celsius.

Example 1 safety testing of prodrug auriculatone sulfate (B)

1.1 safety study test of intraperitoneal injection

Male mice of the Kunming species were randomly divided into 3 groups of 5 mice each. The state of the mice was observed after administration of 50, 100, 200mg/kg B by intraperitoneal injection, respectively.

As a result: no mouse dies in 3 days, and the state of the mouse is normal, which shows that the auriculatone sulfate does not have obvious toxic or side effect at the given dosage.

1.2 safety study test for intravenous injection

Male mice of the Kunming species were randomly divided into 3 groups of 5 mice each. The status of the mice was observed after administration of 25, 5, 1mg/kg B by intravenous injection, respectively.

As a result: no mouse dies in 3 days, and the state of the mouse is normal, which shows that the auriculatone sulfate does not have obvious toxic or side effect at the given dosage.

Example 2 prevention and protection test of prodrug auriculatone sulfate on acute liver injury

2.1 pretreatment of drugs

(1) B: dissolved in 3% Tween 80 to prepare a 2.5mg/ml solution.

(2) APAP: dissolving in 30% PEG400, and making into 15mg/ml solution

2.2 animal treatment

40 Kunming male mice each weighing about 30g were randomly divided into 5 groups of 8 mice each.

(1) Blank group: no treatment is done.

(2) Model group: APAP at 300 mg/kg.

(3) Control group B: 50mg/kg of B.

(4) Group B: the B pretreatment was given first at mg/kg once a day for three consecutive days and APAP at 300mg/kg 2h after the last dose.

(5) APAP is administered at 300mg/kg 2h after 1000mg/kg NAC.

Each group was administered by intraperitoneal injection. Blood is taken through eyeballs 24h after molding, then the neck is removed, the animal is killed, and the liver is taken out for later use. The blood was centrifuged at 3000rpm for 10min at 4 ℃ and then serum was separated. One part of liver is soaked in 4% formaldehyde for pathological examination, and the other part is made into 10% liver homogenate. The kit is used for detecting indexes ALT and AST of liver lesion. The liver is soaked and fixed in 4% formaldehyde solution and then used for pathological tissue examination.

2.3 test results

As can be seen from the above table, the serum levels of ALT and AST increased significantly (p < 0.05) 24h after APAP administration at 300mg/kg, and the pathological section showed that the structure of the liver tissue of the model group (B) had been destroyed, indicating that the liver injury model was successfully replicated. pre-NAC or B treatment was able to counter the ALT and AST increase caused by APAP (p < 0.05), and the administered group was not significantly different from the normal control group (p > 0.05); pathological sections show that the liver structure of NAC or B administration group is basically normal, and has no obvious difference from a normal control group. The auriculatone sulfate can completely protect mice from APAP-induced acute liver injury.

Example 3 treatment and prevention of acute liver injury with the prodrug auriculatone sulfate

3.1 pretreatment of drugs

(1) B: dissolved in 3% Tween 80 to prepare a 2.5mg/ml solution.

(2) APAP: dissolving in 30% PEG400, and making into 15mg/ml solution

2.2 animal treatment

Male Kunming mice 32, each weighing around 30g, were randomly divided into 4 groups of 8 mice each.

(1) Blank group: no treatment is done.

(2) Model group: APAP at 300 mg/kg.

(3) The NAC group was administered 300mg/kg APAP for 1 hour for molding and 1000mg/kg NAC for molding.

(4) Group B was administered with 300mg/kg APAP for 1 hour for molding and 50mg/kg B.

Each group was injected intraperitoneally. The animals were treated 24h after molding and then decapped to sacrifice the animals and remove the liver for use. The blood was centrifuged at 3000rpm for 10min at 4 ℃ and then serum was separated. One part of liver is soaked in 4% formaldehyde for pathological examination, and the other part is made into 10% liver homogenate. The kit is used for detecting indexes ALT and AST of liver lesion.

2.3 test results

From the above test results, it can be seen that the ALT and AST levels in serum significantly increased (p < 0.05) after the APAP of 300mg/kg was administered for modeling, and the pathological section shows that the structure of the liver tissue of the model group (APAP) has been destroyed, indicating that the liver injury model is successfully replicated. In view of the fact that the liver injury is already caused after 1h of APAP modeling, the ALT and AST levels in serum can be remarkably reduced to normal levels (p is less than 0.05) by administering 1000mg/kg of NAC or 50mg/kg of B, which shows that the ALT and AST increases (p is less than 0.05) in serum caused by APAP acute liver injury, the continuous damage of large-dose APAP to the liver is prevented, the protective effect is achieved, and the therapeutic effect on APAP-induced acute liver injury can be achieved.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims (4)

1. A compound of the following general formula (I):

wherein R1 is partial structure of sulfate of the following general formula (V), R7 can be hydrogen or alkyl; r2 is an oxygen atom;

2. the compound of claim 1, wherein the pharmaceutically acceptable salt is a pharmaceutically acceptable salt of the sodium, potassium, calcium, ammonium sulfate ester.

3. A pharmaceutical composition for preventing, protecting and treating liver damage comprising a therapeutically effective amount of a compound of any one of claims 1-2 and a pharmaceutically acceptable carrier.

4. Use of a compound or pharmaceutical composition according to any one of claims 1 to 3 for the preparation of a medicament for the prevention, protection and treatment of liver damage.