Application of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives in the preparation of drugs for the prevention and treatment of polycystic ovary syndrome

Abstract

The present invention belongs to the field of biomedicine, and particularly relates to the use of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for preventing and / or treating polycystic ovary syndrome or their lead compounds. The N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives or pharmaceutically acceptable salts thereof provided by the present invention have the effects of improving polycystic ovary syndrome lesions, normalizing disordered estrous cycles, effectively reducing serum androgen levels, and reducing insulin resistance index, have good bioavailability, and do not have potential cardiotoxicity, and can be used in the development and preparation of new drugs for preventing and / or treating PCOS or their lead compounds.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to the use of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for preventing and / or treating polycystic ovary syndrome or their lead compounds. Background Art

[0002] Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women, characterized by hyperandrogenism, abnormal ovulation, and polycystic ovaries, often accompanied by insulin resistance, glucose intolerance, and obesity. PCOS seriously endangers the reproductive health of women of childbearing age, and also affects the health of offspring during the fetal period, childhood, childbearing period, and even middle-aged and elderly years. However, due to the clinical heterogeneity of PCOS and the complexity of the syndrome, its etiology and pathophysiological mechanism have not yet been elucidated, and there are currently no therapeutic drugs for PCOS at home and abroad. At present, clinical drug treatment methods and their limitations mainly include: 1. For patients in adolescence and childbearing age who have no fertility requirements, oral contraceptives are used to adjust the menstrual cycle, but this method can only cause artificial cycle "menstruation", protect the endometrium stimulated by single estrogen caused by long-term anovulation, and reduce acne and hirsutism caused by hyperandrogenism, but it cannot treat PCOS. Menstrual irregularities and amenorrhea can still occur after stopping the drug; 2. For anti-hyperandrogen treatment with poor contraceptive treatment, spironolactone is often used, but the drug treatment cycle is long, and long-term use of large doses often causes hypotension and hyperkalemia, as well as side effects such as headache or polyuria; 3. For patients with obesity and lipid metabolism disorders, the diagnosis and treatment guidelines recommend the use of metformin or insulin sensitizers for treatment. Long-term use often causes gastrointestinal side effects and vitamin B12 deficiency, as well as aggravated liver and kidney damage and other side effects. In summary, each type of existing drug is for single symptom treatment, hormone drugs have more complications, and other drugs have poor efficacy. Therefore, there is currently a lack of comprehensive drugs that effectively treat PCOS. Summary of the invention

[0003] In order to overcome the above-mentioned shortcomings and deficiencies of the prior art, the primary object of the present invention is to provide an N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof for use in the preparation of a drug or a lead compound thereof for preventing and / or treating polycystic ovary syndrome (PCOS).

[0004] The N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof provided by the present invention has the effects of improving polycystic ovary syndrome (PCOS) lesions, normalizing the disordered estrous cycle, effectively reducing the androgen level in serum, and reducing the insulin resistance index, and can be used to develop and prepare a new drug for treating PCOS.

[0005] Another object of the present invention is to provide a pharmaceutical composition for treating PCOS.

[0006] The purpose of the present invention is achieved through the following solutions:

[0007] Use of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for preventing and / or treating polycystic ovary syndrome (PCOS) or their lead compounds.

[0008] The N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative has a structure as shown in the following formula I:

[0009]

[0010] Wherein, R0 is independently selected from: Halogen, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, -OH, -NO2, -CN; wherein the substitution is one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl and phenyl;

[0011] And at least one R0 is

[0012] n is a positive integer from 1 to 4;

[0013] R1 is selected from: substituted or unsubstituted C1-C10 alkyl; substituted or unsubstituted C3-C10 cycloalkyl; ZY- and R5(R6)N-; wherein the substitution is one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl;

[0014] R5 and R6 are independently selected from hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C1-C10 alkyl-carbonyl, substituted or unsubstituted C1-C10 alkoxy-carbonyl, or substituted or unsubstituted C5-C20 aryl; wherein the substitution is with one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, carboxyl, -OH, -NH2 and phenyl;

[0015] Z is selected from: substituted or unsubstituted C5-C20 aryl, substituted or unsubstituted C3-C20 heteroaryl, and substituted or unsubstituted 5-20 membered heterocyclic group; wherein the substitution is one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl and phenyl;

[0016] Y is a C1-C10 alkylene group which is absent, substituted or unsubstituted, wherein the substitution is a substituent having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, and C2-C4 alkenyl;

[0017] R2 is selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C3-C8 cycloalkyl, -NO2, -CN, -OH, substituted or unsubstituted C1-C6 alkyl-carbonyl, or substituted or unsubstituted C1-C6 alkoxycarbonyl; wherein substituted refers to having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C2-C4 alkenyl and phenyl;

[0018] R3 is selected from the group consisting of substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C5-C20 aryl, R9-C(O)-, cyano, formamide and tetrazole; wherein substitution refers to having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, carboxyl, -OH, -NH2 and phenyl;

[0019] R4 is independently selected from: halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C5-C20 aryl, -NO2, -CN, -OH, R9-C(O)-, R 10 (R 11 )N- or C5-C20 aryl-C(O)-N(R 10 )R 11 -; wherein substituted refers to having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, carboxyl, -OH, -NH2 and phenyl;

[0020] R9 group: hydroxyl, mercapto, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy and R 10 (R 11 )N-; wherein substituted means having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C2-C4 alkenyl and phenyl;

[0021] R 10 and R 11 Each is independently hydrogen, C1-C10 alkyl or C1-C10 haloalkyl;

[0022] m is an integer from 0 to 5.

[0023] Furthermore, the drugs are the same or different and include a therapeutically effective amount of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof.

[0024] Furthermore, the drugs that are the same or different can be made into various pharmaceutical dosage forms by conventional methods, including: tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, hard capsules, soft capsules, oral liquids, lozenges, granules, granules, pills, pills, suspensions, wine preparations, tinctures, drops and other oral dosage forms and injections and other dosage forms other than oral administration, such as injections.

[0025] Furthermore, the drugs, whether the same or different, may also contain one or more pharmaceutically acceptable carriers or excipients.

[0026] Furthermore, the carrier or auxiliary material may include a diluent, a binder, a surfactant, a wetting agent, an adsorption carrier, a lubricant, a filler, a disintegrant, a preservative, and the like.

[0027] The present invention also provides a pharmaceutical composition for treating PCOS, comprising the N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof or other pharmaceutically acceptable components thereof.

[0028] The present invention also provides the use of the above-mentioned N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof in the preparation of a drug for reducing androgen levels.

[0029] The N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof provided by the present invention has the effects of improving polycystic ovary syndrome (PCOS) lesions, normalizing the disordered estrous cycle, effectively reducing the androgen level in serum, and reducing the insulin resistance index, has good bioavailability, and has no potential cardiac toxicity, and can be used in the development and preparation of novel drugs for preventing and / or treating PCOS or their lead compounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0031] Figure 1 This is the result of H&E staining of ovarian morphology observation;

[0032] Figure 2This is the result of ultrasound ovarian morphology observation;

[0033] Figure 3 is the ultrasound ovarian diameter;

[0034] Figure 4 is the ultrasound ovarian area;

[0035] Figure 5 The results of microscopic observation of vaginal secretions of mice;

[0036] Figure 6 This is a statistical chart of the mouse estrous cycle;

[0037] Figure 7 is the serum testosterone level of mice;

[0038] Figure 8 is the insulin resistance level of mice;

[0039] Fig. 9 This is the plasma drug-dose curve of VSP-51-2 injected intravenously into rats;

[0040] Fig.10 This is the plasma drug-dose curve of VSP-51-2 after oral administration to rats. DETAILED DESCRIPTION

[0041] The present invention is further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto. The materials involved in the following examples can be obtained from commercial channels unless otherwise specified. The methods described are conventional methods unless otherwise specified.

[0042] In one embodiment, an N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof is used in the preparation of a drug or a lead compound thereof for preventing and / or treating polycystic ovary syndrome (PCOS).

[0043] In one embodiment, the N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative has a structure as shown in the following formula I:

[0044]

[0045] Wherein, R0 is independently selected from: Halogen, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, -OH, -NO2, -CN; wherein the substitution is one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl and phenyl;

[0046] And at least one R0 is

[0047] n is a positive integer from 1 to 4;

[0048] R1 is selected from: substituted or unsubstituted C1-C10 alkyl; substituted or unsubstituted C3-C10 cycloalkyl; ZY- and R5(R6)N-; wherein the substitution is one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl;

[0049] R5 and R6 are independently selected from hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C1-C10 alkyl-carbonyl, substituted or unsubstituted C1-C10 alkoxy-carbonyl, or substituted or unsubstituted C5-C20 aryl; wherein the substitution is with one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, carboxyl, -OH, -NH2 and phenyl;

[0050] Z is selected from: substituted or unsubstituted C5-C20 aryl, substituted or unsubstituted C3-C20 heteroaryl, and substituted or unsubstituted 5-20 membered heterocyclic group; wherein the substitution is one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl and phenyl;

[0051] Y is a C1-C10 alkylene group which is absent, substituted or unsubstituted, wherein the substitution is a substituent having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, and C2-C4 alkenyl;

[0052] R2 is selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C3-C8 cycloalkyl, -NO2, -CN, -OH, substituted or unsubstituted C1-C6 alkyl-carbonyl, or substituted or unsubstituted C1-C6 alkoxycarbonyl; wherein substituted refers to having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C2-C4 alkenyl and phenyl;

[0053] R3 is selected from the group consisting of substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C5-C20 aryl, R9-C(O)-, cyano, formamide and tetrazole; wherein substitution refers to having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, carboxyl, -OH, -NH2 and phenyl;

[0054] R4 is independently selected from: halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C5-C20 aryl, -NO2, -CN, -OH, R9-C(O)-, R 10 (R 11 )N- or C5-C20 aryl-C(O)-N(R 10 )R 11 -; wherein substituted refers to having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 haloalkyl, carboxyl, -OH, -NH2 and phenyl;

[0055] R9 group: hydroxyl, mercapto, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy and R 10 (R 11 )N-; wherein substituted means having one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, C2-C4 alkenyl and phenyl;

[0056] R 10 and R 11 Each is independently hydrogen, C1-C10 alkyl or C1-C10 haloalkyl;

[0057] m is an integer from 0 to 5.

[0058] In one embodiment, the drugs are the same or different and include a therapeutically effective amount of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof.

[0059] The dosage of the drug or pharmaceutical composition of the present invention can be prescribed in a variety of ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, morbidity, diet, administration time, administration route, excretion rate and reaction sensitivity. A skilled physician can usually easily determine the prescription and the dosage that is effective for the desired treatment or prevention.

[0060] The therapeutically effective amount refers to an amount that can produce a therapeutic effect on humans and / or animals and can be accepted by humans and / or animals. For example, a therapeutically effective amount refers to the amount of the drug required to produce the desired therapeutic effect, and the therapeutic effect can be reflected by the results of clinical trials, model animal studies and / or in vitro studies. The pharmaceutically effective amount depends on several factors, including but not limited to: characteristic factors of the subject to be treated (such as the subject's height, weight, gender, age and medication history, etc.), and the severity of the disease.

[0061] In a specific embodiment of the present invention, the therapeutically effective amount refers to an amount that can produce a therapeutic effect on patients with polycystic ovary syndrome and can be accepted by the patients.

[0062] In a specific embodiment of the present invention, the administration of the drug or pharmaceutical composition includes, but is not limited to, oral administration, parenteral administration, administration by inhalation spray, topical administration, rectal administration, nasal administration, buccal administration, vaginal administration, or administration by implanted drug storage device. Oral administration or injection is preferred.

[0063] In a specific embodiment of the present invention, any orally acceptable dosage form may be used, including but not limited to capsules, tablets, aqueous suspensions or solutions.

[0064] In a specific embodiment of the invention, liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.

[0065] In a specific embodiment of the invention, solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.

[0066] In a specific embodiment of the present invention, the drug or pharmaceutical composition also includes a pharmaceutically acceptable carrier or adjuvant. The pharmaceutically acceptable carrier or adjuvant may contain an inert component that does not unduly inhibit the biological activity of the compound. Pharmaceutically acceptable carriers or adjuvants should be biocompatible, for example, non-toxic, non-inflammatory, non-immunogenic or no other undesirable reactions or side effects when administered to a subject. Standard pharmaceutical preparation techniques can be used.

[0067] Pharmaceutically acceptable carriers or excipients include, but are not limited to, diluents, adhesives, surfactants, wetting agents, adsorption carriers, lubricants, fillers, disintegrants, preservatives, etc. These substances are used to help the stability of the formulation or to help improve the activity or its biological effectiveness or to produce an acceptable taste or smell in the case of oral administration. The preparations that can be used in such pharmaceutical compositions can be in the form of the original compound itself or optionally in the form of a pharmaceutically acceptable salt thereof. The pharmaceutical composition thus formulated can be administered by any appropriate method known to those skilled in the art as needed.

[0068] Among them, the diluent includes but is not limited to lactose, sodium chloride, glucose, urea, starch, and water.

[0069] Binders include, but are not limited to, starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, ethylcellulose, polyvinyl alcohol, polyethylene glycol, polyvinylpyrrolidone, alginic acid and alginates, xanthan gum, hydroxypropyl cellulose and hydroxypropyl methylcellulose.

[0070] Surfactants include, but are not limited to, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, and cetyl alcohol.

[0071] Wetting agents include, but are not limited to, glycerin and starch.

[0072] Adsorption carriers include, but are not limited to, starch, lactose, bentonite, silica gel, kaolin, and bentonite.

[0073] Lubricants include, but are not limited to, zinc stearate, glyceryl monostearate, polyethylene glycol, talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearyl fumarate, polyoxyethylene monostearate, monolauric sucrose ester, sodium lauryl sulfate, magnesium lauryl sulfate, and magnesium dodecyl sulfate.

[0074] Fillers include, but are not limited to, mannitol (granular or powdered), xylitol, sorbitol, maltose, erythrose, microcrystalline cellulose, polymeric sugars, coupling sugars, glucose, lactose, sucrose, dextrin, starch, sodium alginate, laminarin powder, agar powder, calcium carbonate, and sodium bicarbonate.

[0075] Disintegrants include, but are not limited to, cross-linked vinyl pyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropyl methyl, cross-linked sodium carboxymethyl cellulose, and soybean polysaccharides.

[0076] In a specific embodiment of the present invention, the drug or pharmaceutical composition also includes other drugs for treating polycystic ovary syndrome, that is, the N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative can be used in combination with other compounds that can be used to treat polycystic ovary syndrome, and the other drugs for treating polycystic ovary syndrome include but are not limited to clomiphene, letrozole, gonadotropin, etc.

[0077] Example 1: Preparation of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives

[0078] The preparation of 5-(benzylcarbamoyl)-1-(4-chloro-3-fluorobenzyl)-1H-indole-2-carboxylic acid (named VSP-51-2) can refer to CN 105175309A; the specific structural formula is as follows:

[0079]

[0080] Example 2: Anti-polycystic ovary syndrome (PCOS) activity of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives

[0081] (1) Establishment of PCOS model

[0082] C57 mice were raised in the clean-grade experimental animal room of Guangzhou Medical University;

[0083] Letrozole is an aromatase inhibitor that can inhibit the conversion of ovarian androgens to estrogens, thereby causing hyperandrogenism. At the same time, high-fat feeding can cause insulin resistance. Four-week-old C57 mice were fed a high-fat diet and given letrozole (6 mg / kg / day) by gavage for 21 days to create a PCOS model of hyperandrogenism and insulin resistance.

[0084] (2) Experimental grouping and drug administration: This experiment was divided into a normal control group (Control), a model group (PCOS), a positive control metformin group (Metformine), and a VSP-51-2 group; each group was set up with 3 replicates, and each replicate had 5 mice. After successful modeling, the Control and PCOS groups were given an equal amount of PEG300 by gavage every day, the metformin group was given 50 mg / kg metformin (dissolved in 0.2 mL PEG300) by gavage every day, and VSP-51-2 was given 50 mg / kg (dissolved in 0.2 mL PEG300) by gavage every day for 28 days.

[0085] (3) Experimental results testing and data collection

[0086] Starting from the 10th day of administration, vaginal smears were taken and stained daily to observe the estrous cycle, blood glucose was tested every 7 days after fasting, and ultrasound was used to observe the ovarian morphology before sampling on the 28th day after administration. Ovarian tissue was stained with HE by paraffin-embedded sections to observe ovarian morphological changes; serum samples were tested for testosterone and insulin levels by ELISA.

[0087] (4) Results analysis

[0088] ①VSP-51-2 significantly improves polycystic ovarian lesions in PCOS mice

[0089] The core feature of reproductive dysfunction in PCOS patients is follicular development disorder, which is manifested by excessive follicle recruitment, obstructed follicle selection and dominance, multicystic ovarian changes, different degrees of follicles and atretic follicles under the tunica albuginea, vacuolar changes, and follicular development without ovulation.

[0090] Figure 1 The morphology of the ovaries in each group was observed under the H&E staining microscope. Figure 1As shown, multiple follicles and corpora lutea at different developmental stages were observed in the ovaries of mice in the normal control group, and the granulosa cells were complete in morphology and neatly arranged; the ovaries of mice in the PCOS model group were increased in atresia and cystic expansion, no oocytes were seen in the mature follicles, the granulosa cells were loosely arranged, the layers became thinner, some of them fell off, the theca cells proliferated, the oocytes were small, and the corpora lutea were less formed; while in the VSP-51-2 group of the present invention, follicles at different developmental stages were observed, the granulosa cells were tightly arranged, the layers increased, the theca cells were thinner, which was similar to the metformin positive control group, and the morphology was close to the normal control group.

[0091] Figure 2 This is the result of ultrasound ovarian morphology observation; Figure 3 is the ultrasound ovarian diameter; Figure 4 is the ultrasound ovarian area. Figure 2 As shown, a large number of antral follicles can be seen around the central stroma of the ovaries in the PCOS model group compared with the control group, and the area and diameter of the ovarian two-dimensional grayscale image are significantly increased; while the area and diameter of the ovarian two-dimensional grayscale image of the VSP-51-2 group of the present invention are significantly reduced compared with the PCOS group, and are better than the metformin positive control group. Figure 3-Figure 4 It can be seen that the ovarian diameter (1.3-1.9mm) and area (1.5-2.0mm) of the PCOS group were 2 ) were significantly higher than those in the normal control group; while the diameter and area of ​​the VSP-51-2 group of the present invention were significantly decreased, and were better than those of the metformin positive control group and close to those of the normal control group.

[0092] ②VSP-51-2 significantly improves the disordered estrous cycle of PCOS mice

[0093] Figure 5 These are the results of microscopic observation of mouse vaginal secretions. Figure 6 This is a statistical chart of the mouse estrous cycle.

[0094] Depend on Figure 5 It can be seen that in the normal control group (proestrus), spindle-shaped epithelial cells were distributed among the white blood cells, indicating that the estrus period was about to begin. In the PCOS group (during estrus), only a large number of white blood cells were found, and no other cells were found; in the metformin positive control group (metestrus), white blood cells, anucleated epithelial cells, and spindle-shaped epithelial cells were found; and in the VSP-51-2 group of the present invention (estrus), a large number of squamous epithelial cells, anucleated cells, and no white blood cells were found.

[0095] Depend on Figure 6It can be seen that the observation results from the 10th day of drug administration after the successful PCOS modeling showed that the normal control group showed a regular 4-5 day estrous cycle (Proestrus: proestrus, Estrus: estrus, Metestrus: metestrus, Diestrus: diestrus); while the estrous cycle of the PCOS model group was disordered and remained in the diestrus; the metformin positive control group had a recovery cycle compared with the PCOS group; and the estrous cycle of the VSP-51-2 group of the present invention recovered to a normal level, which was significantly better than the metformin positive control group.

[0096] ③VSP-51-2 significantly reduces hyperandrogenism in PCOS mice

[0097] Hyperandrogenism is the main clinical manifestation and detection index of PCOS. The present application used ELISA method to detect serum testosterone levels in each group according to the kit instructions (Finetest, Wuhan). Figure 7 is the serum testosterone level of mice.

[0098] As shown in the figure, the testosterone content in the serum of the normal control group is 2.1-5.0ng / mL; the serum testosterone content of the PCOS group is 10.1-19.0ng / mL, which is significantly higher than that of the normal control group; the serum testosterone concentration of the metformin positive control group is 3.7-7.7ng / mL, and the concentration level is significantly lower than that of the PCOS group; and the serum testosterone content of the VSP-51-2 group of the present invention is 3.0-5.0ng / mL, which is lower than that of the metformin positive control group and closer to the level of the normal control group. Therefore, the VSP-51-2 of the present invention significantly reduces hyperandrogenemia in PCOS mice, and the effect is significantly better than that of the positive control drug metformin.

[0099] ④VSP-51-2 significantly reduces insulin resistance in PCOS mice

[0100] Insulin resistance is another major clinical manifestation and detection index of PCOS. Clinically, the degree of insulin resistance is mainly determined by calculating the Homeostatic Model Assessment for Insulin Resistance (HOMA-IR). The specific calculation formula is: HOMA-IR = fasting blood glucose (mmol / L) × fasting insulin (mIU / L) / 22.5. Mice in different groups began to fast and drink water on the evening of the 27th day of treatment. Blood was collected from the tip of the tail after 12 hours, and fasting blood glucose was tested using blood glucose. The serum insulin level was detected by ELISA, and the specific operation was carried out according to the instructions of the ELISA kit (Shanghai ELISA Biological, Shanghai). Figure 8 is the insulin resistance level of mice.

[0101] As can be seen from the figure, the HOMA-IR of the normal control group is 0.20-0.26; the PCOS group significantly increased to 0.28-0.36; while the HOMA-IR of the VSP-51-2 group of the present invention and the positive control metformin group were significantly reduced, close to the normal control group. This shows that the VSP-51-2 of the present invention can significantly reduce the insulin resistance index of PCOS mice, and its effect is equivalent to that of the positive control drug metformin.

[0102] Example 3: Pharmacokinetic properties of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives

[0103] The pharmacokinetic properties of drugs are important indicators for evaluating drug potential and safety in the development of new drugs. The present invention studies the pharmacokinetic parameters of VSP-51-2 by intravenous injection and oral gavage in rats. SD rats (about 200g) were fasted for 12 hours and randomly divided into two groups (one group of 4 and one group of 3), of which 4 were given 20 mg / kg of VSP-51-2 by oral gavage, and the remaining 3 were given 5 mg / kg of VSP-51-2 by intravenous injection. After administration, 0.5 mL of blood was collected from the fundus vein at 0.1, 0.5, 1, 2, 4, 6, and 8 hours respectively. The blood samples were centrifuged at 8000 r / min for 5 minutes, and the separated plasma samples were stored at -20°C for testing. Take 0.2 mL of plasma, add 0.6 mL of acetonitrile, vortex mix for 1 minute, centrifuge at 12000 r / min for 10 minutes, and take 20 μL of the supernatant for HPLC analysis. The results are shown in Tables 1-2 and Figure 9-10 .

[0104] Table 1 is a statistical table of pharmacokinetic parameters of VSP-51-2 injected intravenously into rats; Table 2 is a statistical table of pharmacokinetic and blood parameters of VSP-51-2 injected intravenously into rats; Fig. 9 This is the plasma drug-dose curve of VSP-51-2 injected intravenously into rats; Fig.10 This is the plasma drug-dose curve of VSP-51-2 after oral administration to rats.

[0105] Table 1

[0106]

[0107] Table 2

[0108]

[0109] The results showed that after oral administration of VSP-51-2, the blood drug concentration reached a peak value (7937±2110ng / mL) at 0.25h, the drug half-life was 1.37±0.228h, the average residence time was 1.8±0.357h, and the absolute bioavailability was about 21%. The above data show that after oral administration, VSP-51-2 of the present invention can be rapidly absorbed and enter the circulation, and the absolute bioavailability of oral administration is greater than 20%, reaching an acceptable bioavailability.

[0110] Example 4: hERG Inhibitory Effects of N-Benzyl-5 / 6-Formylaminoindole-2-Carboxylic Acid Derivatives

[0111] Since some non-cardiovascular drugs have been found to be cardiotoxic, the cardiotoxicity of drugs is one of the key indicators of drug safety evaluation. In myocardial cells, the inhibition of potassium ion channels encoded by hERG (human Ether-a-go-go Related gene) is an important mechanism leading to cardiotoxicity. Therefore, the present invention detects the inhibitory effect of compounds on hERG potassium channels.

[0112] Different concentrations (0.0128, 0.064, 0.32, 1.6, 8, 40 μM) of VSP-51-2 compounds were added to stably transfected hERG-HEK293 cells, and Cisapride was used as a positive control. The hERG K+ channel current was recorded using the Nanion Patchliner 8-channel fully automated patch clamp technique (patch clamp settings: Patchmaster software was used to collect and store hERG potassium channel current (Ikr) data on a computer through an EPC-10 amplifier), and the hERG current inhibition under different concentrations was calculated.

[0113] The hERG inhibition rate (IC 50 ) was greater than 40 μM, and the hERG inhibition rate of VSP-51-2 at 40 μM was less than 20%, indicating that VSP-51-2 had no potential cardiotoxicity.

[0114] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Use of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives or pharmaceutically acceptable salts thereof in the preparation of a drug for preventing and / or treating polycystic ovary syndrome, characterized in that The N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative has a structure as shown in the following formula I: 。 2. The use according to claim 1, characterized in that: The invention relates to the use of the N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof in the preparation of a drug for reducing androgen levels.

3. Use of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives or pharmaceutically acceptable salts thereof in the preparation of a drug for improving PCOS polycystic ovary syndrome, characterized in that The N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative has a structure as shown in the following formula I: 。 4. Use of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives or pharmaceutically acceptable salts thereof in the preparation of a drug for improving PCOS insulin resistance, characterized in that The N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative has a structure as shown in the following formula I: 。 5. Use of N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivatives or pharmaceutically acceptable salts thereof in the preparation of a drug for improving PCOS physiological cycle disorders, characterized in that The N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative has a structure as shown in the following formula I: 。 6. The use according to any one of claims 1 to 5, characterized in that: The drug comprises a therapeutically effective amount of an N-benzyl-5 / 6-formylaminoindole-2-carboxylic acid derivative or a pharmaceutically acceptable salt thereof.

7. The use according to any one of claims 1 to 5, characterized in that: The medicine is prepared into various pharmaceutical dosage forms by conventional methods, and these dosage forms include tablets, capsules, oral liquids, lozenges, granules, granules, pills, pills, suspensions, wine preparations, tinctures, drops and injections.

8. The use according to any one of claims 1 to 5, characterized in that: The medicine also contains one or more pharmaceutically acceptable carriers or excipients.

9. The use according to claim 8, characterized in that: The carrier or auxiliary material includes at least one of a diluent, a binder, a surfactant, a wetting agent, an adsorption carrier, a lubricant, a disintegrant, and a preservative.

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