CN117417303B - Medicine for treating endometriosis and preparation method thereof - Google Patents

Abstract

The invention relates to a medicine for treating endometriosis and a preparation method thereof. The medicine has a structure shown in a formula (I). The medicine has good endometriosis treatment effect, is superior to traditional Chinese medicine active monomers such as 6-shogaol, and can be expected to be developed into an effective endometriosis treatment medicine.

Description

Medicine for treating endometriosis and preparation method thereof

Technical Field

The invention relates to the field of medicines, in particular to a medicine for treating endometriosis, and also relates to a preparation method of the medicine.

Background

Endometriosis (abbreviated as "endo-dysbiosis") is a gynecological disease, which is more common in women of childbearing age, and is a benign disease, but has similarities to tumors. The histological features are that the endometrium-like gland and matrix exist and grow in other parts outside the uterine cavity, the common parts are the ovary, the ligament, the abdominal cavity and the like, and serious patients can be ectopic to intestinal tracts, urinary bladder and even other viscera. Endometriosis is classified into ovarian endometriosis, superficial peritoneal endometriosis, deep invasive endometriosis and the like according to the position of the lesions. Endometriosis can periodically proliferate and destroy, much like endometrium. In most cases, endometriosis is more closely related to dysmenorrhea, dyspareunia and pelvic pain and can severely affect the physical and mental health of the affected female. Meanwhile, the internal heteropathy also increases the probability of infertility of the sick female. The occurrence of internal complications can seriously affect the physical and mental health of the affected female, and cause great burden and pressure to the individuals, families and society.

Surgical treatment has been considered as the final treatment for endometriosis, but recent studies have shown that surgical treatment is not effective and patients require long-term treatment. Surgical treatment requires consideration of the risk of occurrence of complications of the urinary system, intestinal tract, blood vessels, and nervous system, and pain may recur or persist in cases where the excision of lesions is incomplete. Common drugs for endometriosis include oral contraceptive drugs, progestogens, progestogen receptor antagonists, androgen derivatives, gonadotrophin releasing hormone agonists (GnRHa), gonadotrophin releasing hormone antagonists and the like, wherein the purpose of the contraceptive is to reduce pituitary gonadotrophin levels and act directly on the endometrium and ectopic endometrium resulting in endometrium atrophy and reduced menstrual flow; progestogen causes "amenorrhea" and "pseudopregnancy" by a series of actions, and progestogen receptor antagonists have a potent antiprogestogenic action, causing amenorrhea to atrophy the lesions. GnRH-a causes a hypoestrogenic state in the body by reducing the sex hormones secreted by the ovaries, and temporary amenorrhea occurs. However, these drugs have potential adverse reactions when used for a long period of time, and particularly affect the estrogenic environment of women of childbearing age. The key of the aim of long-time control of endometriosis is to relieve and eliminate pain, protect fertility, reduce recurrence rate, discover malignant lesions as early as possible and realize the life value of patients. Therefore, a medicine with more definite curative effect, good tolerance and higher cost performance is sought, and the medicine is a development route for comprehensively and individually treating endometriosis.

Traditional Chinese medicine is an effective means for treating endometriosis, but the previous research is focused on traditional Chinese medicine compounds. With the development of modern pharmacological research and clinical research, a number of traditional Chinese medicine monomer components capable of effectively relieving and treating endometriosis have been found, such as epimedium herb, danshensu, salvianolic acid, luteolin, apigenin, nobiletin, baicalein, genistein, curcumin, sinomenine, camptothecine, vinblastine, dioscin, ginsenoside, imperatorin, andrographolide, parthenolide, costunolide, ligustrazine, glycyrrhizic acid, 6-gingerol, garlic derivatives and the like. However, these traditional Chinese medicine components have the problem of insufficient curative effect, but by taking them as parent compounds, more endometriosis therapeutic drugs with better curative effect can be developed.

Disclosure of Invention

Aiming at the problems existing in the prior art, the invention provides an endometriosis treatment drug with better curative effect, which starts from known traditional Chinese medicine active monomers with endometriosis treatment activity.

In one aspect, the present invention provides a compound of formula (I), a pharmaceutically acceptable salt, stereoisomer, prodrug thereof:

In formula (I), R ₁、R₂, which are identical or different, each independently represent hydrogen, substituted or unsubstituted (C1-6) alkyl;

r ₃、R₄、R₅ are identical or different and each independently represents hydrogen, hydroxy, mercapto, amino, substituted or unsubstituted (C1-6) alkoxy.

In a preferred embodiment, said "substituted or unsubstituted" means unsubstituted or mono-or polysubstituted with groups selected from hydroxy, (C1-4) alkoxy, (C1-6) alkyl, halogen.

In a preferred embodiment, R ₁、R₂ are the same or different and each independently represents hydrogen, methyl.

In a preferred embodiment, R ₁ represents methyl and R ₂ represents hydrogen.

In a preferred embodiment, R ₃、R₄、R₅, which are the same or different, each independently represent hydrogen, hydroxy, methoxy, and at least one is selected from hydroxy, methoxy.

In a preferred embodiment, R ₄ represents hydroxy, methoxy, R ₃、R₅ are the same or different and each independently represents hydrogen, hydroxy, methoxy.

In a preferred embodiment, R ₃、R₄ are the same or different and each independently represent hydroxy, methoxy, R ₅ represents hydrogen, hydroxy, methoxy.

In a preferred embodiment, R ₃、R₄、R₅ represents hydroxy, methoxy.

In a more preferred embodiment, R ₃、R₄、R₅ represents hydroxy.

In a preferred embodiment, the compound of formula (I) is selected from:

in the present invention, halogen means fluorine, chlorine, bromine or iodine.

In the present invention, alkyl represents a straight-chain or branched saturated hydrocarbon group, preferably having 1 to 10 carbon atoms, more preferably having 1 to 6 or 1 to 4 carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, and the like.

Pharmaceutically acceptable salts of the compounds of formula (I) according to the invention are salts derived from inorganic or organic acids. Such acid addition salts include the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorite, cyclopentane propionate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, hemisulfate, caproate, hydrochloride, hydrobromide, hydroiodide, lactate, maleate, methanesulfonate, nicotinate, oxalate, persulfate, picrate, pivalate, propionate, succinate, tartrate, tosylate, undecanoate, and the like, but are not limited thereto.

In the present invention, the term "prodrug" means a compound that is capable of being converted in vivo into a compound of formula (I) according to the present invention. Such conversion is effected by hydrolysis of the prodrug in the blood or enzymatic conversion to the parent structure in the blood or tissue. The prodrug of the invention can be ester, and in the prior invention, the ester can be phenyl ester, aliphatic ester, acyloxymethyl ester, carbonic ester, carbamate and amino acid ester as the prodrug.

The present invention provides pharmaceutical compositions suitable for pharmaceutical use comprising at least one compound of formula (I) as described herein, pharmaceutically acceptable salts, stereoisomers, prodrugs thereof. The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle, or combination thereof.

The present invention may be used to administer the compounds and compositions in any convenient form of administration, e.g., tablets, powders, capsules, solutions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches and the like. Such compositions may contain the usual components of pharmaceutical preparations, such as carriers, diluents, excipients, pH modifying agents, sweeteners, fillers and other active agents.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), or with the following ingredients: (a) Fillers or compatibilizers, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) Binders, for example, hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) Disintegrants, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (d) Wetting agents, such as cetyl alcohol and glycerol monostearate; and (f) a lubricant, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents. Formulations for parenteral injection may comprise physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

For adult patients, the compounds of the present invention can be administered orally or parenterally in an amount of 0.001 to 500mg as 1 administration, 1 time per day or divided into several times. The administration amount may be appropriately increased or decreased depending on the type of disease to be treated, the age, weight, symptoms, and the like of the patient.

The pharmaceutical compositions of the compounds of formula (I), pharmaceutically acceptable salts, stereoisomers, prodrugs thereof, described herein, may be used alone or in combination with other therapeutic agents. Combination therapy may provide additive or synergistic effects, advantageously to provide synergistic effects.

Preferably, the other therapeutic agent is selected from other drugs useful for the treatment of endometriosis, such as contraceptive drugs, progestins, progestin receptor antagonists, androgen derivatives, gonadotrophin releasing hormone agonists (GnRHa), gonadotrophin releasing hormone antagonists and the like, especially GnRHa.

The combination therapy may be administered in a simultaneous or sequential regimen. When administered sequentially, the combination may be administered in two or more applications. The compounds may be administered together in a single pharmaceutical combination, or separately, and when administered separately, may be performed simultaneously or sequentially in any order.

Accordingly, the present invention also provides a combination comprising at least one compound of formula (I) according to the invention, pharmaceutically acceptable salts, stereoisomers, prodrugs thereof and other medicaments useful for the treatment of endometriosis.

In a further aspect, the present invention provides the use of a compound of formula (I), a pharmaceutically acceptable salt, stereoisomer, prodrug thereof, or a pharmaceutical composition according to the invention, or a combination according to the invention, in the manufacture of a medicament for the treatment of endometriosis.

Preferably, the endometriosis comprises ovarian endometriosis, peritoneal endometriosis, deep invasive endometriosis, preferably peritoneal endometriosis.

In a further aspect, the present invention provides a process for the preparation of a compound of formula (I), comprising the steps of:

Wherein R ₁-R₅ is as described herein before;

X is selected from halogen, preferably chlorine, bromine, iodine;

step 1: condensing a compound of formula a and a compound of formula b to form a compound of formula c;

Step 2: the compound of formula c is hydro-converted to the compound of formula (I).

Preferably, step 1 is performed in the presence of an organic base, a phosphine ligand, and a palladium catalyst, wherein the organic base is at least one selected from triethylamine and pyridine; the phosphine ligand is selected from at least one of triphenylphosphine and tri-tert-butylphosphine; the palladium catalyst is at least one selected from palladium acetate and palladium chloride.

Preferably, the hydrogenation of step 2 comprises any double bond hydrogenation process known in the art, such as catalytic hydrogenation in the presence of metals such as nickel, palladium, platinum, etc. More preferably, the hydrogenation catalyst is palladium on carbon.

Advantageous effects

The invention provides a medicine for treating endometriosis and a preparation method thereof. The medicine of the invention has good endometriosis treatment effect and is superior to traditional Chinese medicine active monomers such as 6-shogaol, and can be expected to be developed into an effective endometriosis treatment medicine.

Detailed Description

The present invention is described in more detail below to facilitate an understanding of the present invention.

The experimental methods in the following examples are conventional methods unless otherwise specified. The specific techniques or conditions are not identified in the examples and are described in the literature in this field or are carried out in accordance with the product specifications.

Example 1:

under the protection of nitrogen, the raw material compounds 1-a (20 mmol,4.10 g) and 1-b (20 mmol,2.96 g) are dissolved in acetonitrile (100 ml), stirred to be completely dissolved, and then triethylamine (3 ml), triphenylphosphine (6 mmol,1.57 g) and palladium acetate (0.22 g) are added and heated to reflux for 6h; after the reaction was completed, the reaction mixture was cooled to room temperature naturally, then, a saturated aqueous sodium hydrogencarbonate solution (80 ml) was added, after thorough mixing, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, concentrated to dryness, and the crude product was recrystallized from anhydrous ethanol to give off-white intermediate 1-c (4.85 g,17.8 mmol) in 89% yield. ESI-MS:273.2[ M+H ] ⁺

Intermediate 1-c (2.72 g,10 mmol) was dissolved in anhydrous diethyl ether (50 ml), 5% palladium on carbon catalyst (0.2 g) was added, and hydrogen was introduced and stirred at room temperature and pressure for 12h. Palladium on carbon was removed by filtration and the reaction was concentrated to dryness and the crude product was purified by a volume ratio of 2:1, to give 2.47g of a white solid compound 1 in a yield of 90%. ESI-MS:275.3[ M+H ] ⁺; elemental analysis: theoretical values C ₁₄H₁₄N₂O₄, C,61.31; h,5.15; n,10.21; o,23.33; actual measurement value ,C,61.21;H,5.16;N,10.23;O,23.34.¹H NMR(400MHz,DMSO-d₆)δ10.01(s,1H),9.78(s,1H),7.80(d,J＝8.9Hz,2H),7.23(s,1H),6.90(d,J＝8.9Hz,2H),4.07(s,3H),3.23(d,J＝8.4Hz,2H),3.06(t,J＝8.4Hz,2H).¹³C NMR(100MHz,DMSO-d₆)δ199.83,161.41,150.42,148.51,147.34,133.55,130.38,130.38,129.84,117.90,117.90,54.57,38.18,30.67.

Example 2

Under the protection of nitrogen, the raw material compounds 1-a (20 mmol,4.10 g) and 2-b (20 mmol,3.28 g) are dissolved in acetonitrile (100 ml), stirred to be completely dissolved, and then triethylamine (3 ml), triphenylphosphine (6 mmol,1.57 g) and palladium acetate (0.22 g) are added and heated to reflux reaction for 5h; after the reaction, the mixture was cooled to room temperature naturally, then saturated aqueous sodium hydrogencarbonate (80 ml) was added, and after thorough mixing, the mixture was extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, concentrated to dryness, and the crude product was recrystallized from anhydrous ethanol to give off-white intermediate 2-c (4.90 g,17.0 mmol) in 85% yield. ESI-MS:289.1[ M+H ] ⁺

Intermediate 2-c (2.88 g,10 mmol) was dissolved in anhydrous diethyl ether (50 ml), 5% palladium on carbon catalyst (0.2 g) was added, and hydrogen was introduced and stirred at room temperature and pressure for 12h. Palladium on carbon was removed by filtration and the reaction was concentrated to dryness, the crude product was purified by a volume ratio of 3:1 to obtain 2.67g of off-white solid compound 2 in 93% yield. ESI-MS:291.2[ M+H ] ⁺; elemental analysis: theoretical values C ₁₄H₁₄N₂O₅, C,57.93; h,4.86; n,9.65; o,27.56; actual measurement value ,C,57.90;H,4.87;N,9.68;O,27.52.¹H NMR(400MHz,DMSO-d₆)δ9.78(s,1H),9.39(s,1H),9.23(s,1H),7.42(d,J＝9.2,1H),7.35(s,1H),7.23(s,1H),6.88(d,J＝9.2Hz,1H),4.07(s,3H),3.24(d,J＝8.4Hz,2H),3.06(t,J＝8.4Hz,2H).¹³C NMR(100MHz,DMSO-d₆)δ199.02,150.60,150.42,148.51,147.34,145.89,133.55,129.86,122.28,115.79,114.94,54.57,37.86,30.70.

Example 3

Under the protection of nitrogen, the raw material compounds 1-a (20 mmol,4.10 g) and 3-b (20 mmol,3.60 g) are dissolved in acetonitrile (100 ml), stirred to be completely dissolved, and then triethylamine (3 ml), triphenylphosphine (6 mmol,1.57 g) and palladium acetate (0.22 g) are added and heated to reflux reaction for 4.5h; after the reaction was completed, the reaction mixture was cooled to room temperature naturally, then, a saturated aqueous sodium hydrogencarbonate solution (80 ml) was added, after thorough mixing, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, concentrated to dryness, and the crude product was recrystallized from anhydrous methanol to give pale yellow intermediate 3-c (5.05 g,16.6 mmol) in 83% yield. ESI-MS:305.3[ M+H ] ⁺

Intermediate 3-c (3.04 g,10 mmol) was dissolved in anhydrous diethyl ether (50 ml), 5% palladium on carbon catalyst (0.2 g) was added, and hydrogen was introduced and stirred at room temperature and pressure for 12h. Palladium on carbon was removed by filtration and the reaction was concentrated to dryness and the crude product was purified by a volume ratio of 2:1 in anhydrous methanol/hexane, 2.88g of a pale yellow solid compound 3 was obtained in a yield of 95%. ESI-MS:307.1[ M+H ] ⁺; elemental analysis: theoretical values C ₁₄H₁₄N₂O₆, C,54.90; h,4.61; n,9.15; o,31.34; actual measurement value ,C,54.92;H,4.63;N,9.10;O,31.33.¹H NMR(400MHz,DMSO-d₆)δ9.78(s,1H),8.74(s,2H),8.39(s,1H),7.23(s,1H),6.97(s,2H),4.07(s,3H),3.23(d,J＝8.4Hz,2H),3.06(t,J＝8.4Hz,2H).¹³C NMR(100MHz,DMSO-d₆)δ199.75,150.42,148.51,147.38,147.34,147.34,138.88,133.55,130.05,107.80,107.80,54.57,37.85,30.67.

Pharmacologically active examples:

Female SD rats with sexual maturity were taken, 40 rats with ages of about 6-8 weeks, and body weight of 200+ -20 g (purchased from Shenyang Ai Kesai Sichuan Biotech Co.). The feeding conditions are as follows: standard illumination (12 h illumination, 12h night environment), the temperature in the feeding room is controlled at 18-22 ℃, the relative humidity is controlled at 40-50%, and standard feed and drinking water are freely eaten for 1 week.

An autograft method is adopted to build a rat endometriosis model. The rats were given intramuscular estradiol at 0.1g/g 1 day before modeling. Daily spot observations were made to determine estrus cycle. The estrus cycle of rats was established based on keratinocyte count in vaginal shed cells, i.e. keratinocytes in vaginal smears were observed under a microscope, estrus when the number of synucleless keratinocytes reached 80%. Modeling surgery was performed after at least 4 estrus cycles of continuous observation and detection.

During operation modeling, 2% ketamine hydrochloride (0.4 ml/100 g) is used for abdominal anesthesia, the incision in the middle of the lower abdomen is selected after shearing and disinfection, the skin is cut at a position 1cm above the pubic symphysis of the rat, and the incision is about 3cm long. The left uterus of the rat was probed and isolated with a separating forceps, and the periuterine vessels were separated and the silk ligature was performed. Then ligate the two ends of the uterine tissue to be excised with silk thread, and cut the left molecular uterine tissue about 1cm long with scissors. It is rapidly placed in physiological saline, the uterus is incised by the wale, the endometrium and the myometrium are separated, and the serous myometrium is separated from the endometrium in an effort. The separated endometrial tissue was then sheared into 0.5cm x 0.5cm sized pieces. The suture lines of 5/0 are used for fixing 1 needle at each of the 4 corners of the endometrium segment on the right side abdominal wall of the incision, and the abdominal cavity is filled with 0.2ml of gentamicin, and the abdominal wall and the skin are sutured layer by layer.

Rats were given a total of 3 injections of estradiol benzoate 100 μg/kg, 1 time 4 days, beginning on day 2 post-surgery. After modeling for 4 weeks, nodes with different sizes are touched on the abdominal wall of the rat, and then the abdominal opening is carried out again according to the first operation method, so that the transparent cystic tumor with circular or oval endometriosis focus can be seen, and the liquid in the capsule is clear and light yellow or light red. The length, width and height values of the endometriosis focus are accurately measured, the endometriosis cyst volume is calculated, and the calculation formula is as follows: length x width x height x 0.5. 2 rats died during the modeling process.

SD rats successfully modeled were randomly divided into 4 groups: model group (7), compound 1 group (8), compound 2 group (8), compound 3 group (8), positive control group (7), and the rats of the administration group are respectively administered with compound 1-3 by stomach infusion every day, and the dosage is 50mg/kg; the positive control group is irrigated with 6-shogaol with the dosage of 50mg/kg; the model group was given an equal amount of double distilled water. The experimental animals were sacrificed within 48 hours after the last administration, the length, width, and height values of the endometriotic lesions were measured again, and the endometriotic cyst volume was calculated.

Statistical analysis adopts SPSS22.0 software to carry out statistical analysis, and data adopts mean value +/-standard deviation ± standard deviation%) Indicating that there is a significant difference when P < 0.05. The results are shown in Table 1 below.

Table 1: therapeutic effects of different drugs on rat endometriosis

From the experimental results, it can be seen that the size of the ectopic focus of the rat model of endometriosis before administration was similar, and the difference between the groups was not statistically significant (P > 0.05). And after the compound 1-3 is given, the ectopic focus volume of the rat in the administration group is obviously reduced, and the difference from the model group has statistical significance (P < 0.05), which indicates that the compound has good treatment effect on endometriosis. Meanwhile, compared with a positive control group to which 6-shogaol is administered, the compound of the invention can better reduce the volume of ectopic foci, and the difference has statistical significance (P < 0.05).

The foregoing is merely a preferred embodiment of the present invention, and it should be noted that modifications and additions may be made to those skilled in the art without departing from the method of the present invention, which modifications and additions are also to be considered as within the scope of the present invention.

Claims (9)

1. A compound of formula (I), a pharmaceutically acceptable salt thereof:

In formula (I), R ₁ represents methyl, R ₂ represents hydrogen;

R ₄ represents a hydroxyl group, and R ₃、R₅ are the same or different and each independently represents hydrogen or a hydroxyl group.

2. The compound of formula (I), pharmaceutically acceptable salts thereof, according to claim 1, wherein the compound of formula (I) is selected from:

3. A pharmaceutical composition comprising at least one compound of formula (I), a pharmaceutically acceptable salt thereof, according to claim 1, and a pharmaceutically acceptable carrier.

4. A pharmaceutical composition comprising at least one compound of formula (I), a pharmaceutically acceptable salt thereof, according to claim 1, and a pharmaceutically acceptable excipient.

5. A combination comprising at least one compound of formula (I), a pharmaceutically acceptable salt thereof, and a further medicament useful for the treatment of endometriosis.

6. Use of a compound of formula (I), a pharmaceutically acceptable salt thereof, according to claim 1, or a pharmaceutical composition according to claim 3 or 4, or a combination medicament according to claim 5, for the manufacture of a medicament for the treatment of endometriosis.

7. The use according to claim 6, wherein the endometriosis comprises ovarian endometriosis, peritoneal endometriosis, deep invasive endometriosis.

8. A process for preparing a compound of formula (I) as claimed in claim 1, comprising the steps of:

wherein R ₁-R₅ is as described in claim 1;

X is selected from halogen;

step 1: condensing a compound of formula a and a compound of formula b to form a compound of formula c;

Step 2: the compound of formula c is hydro-converted to the compound of formula (I).

9. The method of claim 8, wherein X is selected from the group consisting of chlorine, bromine, and iodine.