WO2020248630A1

WO2020248630A1 - Use of compound in preparation of drug

Info

Publication number: WO2020248630A1
Application number: PCT/CN2020/077426
Authority: WO
Inventors: 刘兴国; 杨亮; 林晓冰
Original assignee: 中国科学院广州生物医药与健康研究院
Priority date: 2019-06-13
Filing date: 2020-03-02
Publication date: 2020-12-17
Also published as: CN111617093A

Abstract

The present invention relates to the use of a compound in the preparation of a drug, specifically proposing the use of the compound of formula I or a stereoisomer, nitrogen oxide, solvate, metabolite, or pharmaceutically acceptable salt of the compound of formula I, or a prodrug or derivative thereof in the preparation of a drug, the drug being used for the treatment or prevention of female infertility.

Description

Use of compound in preparing medicine

Technical field

The present invention relates to the field of biomedicine. In particular, the present invention relates to the use of compounds in the preparation of medicines, the use of the compounds in the preparation of kits, the use of the composition in the preparation of pharmaceutical compositions, and the use of the composition in the preparation of health products. use.

Background technique

In recent years, the phenomenon of infertility and low fertility has increased. The most likely explanation is the rapid changes in the environment. At present, more and more people believe that mitochondria play an important role in infertility caused by age and environment. Through autologous mitochondrial injection therapy, mitochondrial transplantation has been used to improve the fertility of women with poor congenital fertility. However, this method is complicated and costly, and cannot play a preventive role.

Therefore, there is an urgent need for drugs to prevent and treat female infertility or fertility difficulties on the market.

Summary of the invention

This application is based on the inventor's discovery and understanding of the following facts and problems:

The inventor found through mouse experiments that the compound represented by formula I can significantly improve the fertility of infertile or fertile mice, especially the fertility of infertile or fertile mice caused by mtDNA mutations, showing that formula I The indicated compounds have great application value in the field of reproduction.

For this reason, in the first aspect of the present invention, the present invention proposes a compound represented by formula I or a stereoisomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt, and its Use of the prodrugs or derivatives thereof in the preparation of medicaments for the treatment or prevention of female infertility,

among them:

Each R is independently H, D, F, Cl, Br, I, OH, NH ₂ , NO ₂ , CN, N ₃ or C _1-6 alkyl;

n is 0, 1, 2, 3, or 4.

It should be noted that the treatment or prevention of "female infertility" needs to be understood in a broad sense, not only refers to the treatment or prevention of women who cannot become pregnant or infertile, but also refers to the treatment or prevention of women who have difficulty in pregnancy or childbirth. In addition, senescence is not directly related to fertility. Senescence is only a cause of infertility and not a necessary condition. Infertility may be caused by aging or other environmental factors. Therefore, whether anti-aging drugs can treat or prevent infertility is completely unknown. The inventor surprisingly discovered that the compounds according to the embodiments of the present invention can effectively treat or prevent female infertility.

According to the embodiment of the present invention, the above-mentioned use may further include at least one of the following additional technical features:

According to an embodiment of the present invention, each R is independently H, D, F, Cl, Br, I, OH, NH ₂ , NO ₂ , CN, N ₃ or C _1-4 alkyl.

According to an embodiment of the present invention, each R is independently H, D, F, Cl, Br, I, OH, NH ₂ , NO ₂ , CN, N ₃ , methyl, ethyl, n-propyl, isopropyl , N-butyl, isobutyl or tert-butyl.

According to an embodiment of the present invention, the compound has the following structure:

According to an embodiment of the present invention, the derivative of the compound is NADH or NAD ⁺ .

According to an embodiment of the present invention, the female infertility is caused by mtDNA mutations. Wherein, the mtDNA mutations include point mutations and deletion mutations. It should be noted that the mtDNA mutation may be caused by aging or other environmental factors. The inventors discovered for the first time that mitochondrial DNA mutations can cause a significant decrease in fertility. At the same time, the inventors were surprised to find that NMN can reverse the decline in fertility caused by mitochondrial DNA mutations and play a significant improvement effect. Therefore, the compounds according to the embodiments of the present invention have a significant effect on female infertility caused by mtDNA mutations.

According to an embodiment of the present invention, the female infertility is caused by point mutations in mtDNA of egg cells. The inventors found that the compounds according to the embodiments of the present invention can effectively treat or prevent female infertility caused by mtDNA point mutations in egg cells.

According to an embodiment of the present invention, the female infertility is caused by an increase in mtDNA point mutations in egg cells. The inventors found that the compounds according to the embodiments of the present invention can effectively treat or prevent female infertility caused by elevated mtDNA point mutations in egg cells.

In the second aspect of the present invention, the present invention proposes the compound represented by formula I or the stereoisomers, nitrogen oxides, solvates, metabolites, pharmaceutically acceptable salts, and prodrugs of the compounds represented by formula I Or the use of derivatives thereof in the preparation of kits for restoring mtDNA mutations or reducing the rate of mtDNA point mutations,

among them:

n is 0, 1, 2, 3, or 4.

The inventor found that the compounds according to the embodiments of the present invention can effectively restore mtDNA mutations or reduce the rate of mtDNA point mutations. Furthermore, the kits prepared according to the compounds of the embodiments of the present invention can effectively restore mtDNA mutations or reduce the rate of mtDNA point mutations, so that For scientific research.

In the third aspect of the present invention, the present invention proposes the use of the composition in the preparation of a pharmaceutical composition for the treatment or prevention of female infertility, and the composition includes the aforementioned compound.

According to an embodiment of the present invention, the composition further includes: a pharmaceutically acceptable excipient, carrier, adjuvant, vehicle, or a combination thereof.

According to an embodiment of the present invention, the composition further includes: other drugs for treating or preventing female infertility.

In the fourth aspect of the present invention, the present invention proposes the use of the composition in the preparation of health products for the prevention of female infertility, and the composition includes the compounds described above.

Description of the drawings

Figure 1 is a schematic diagram of the fertility test results of POLG mutant mice according to an embodiment of the present invention;

Fig. 2 is a schematic diagram of the detection result of NMN improving the fertility of POLG mutant mice according to an embodiment of the present invention.

Detailed description of the invention

The embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary, and are intended to explain the present invention, but should not be construed as limiting the present invention.

It should be noted that, in each part of this specification, the substituents of the compounds disclosed in the present invention are disclosed according to group types or ranges. In particular, the present invention includes each independent sub-combination of each member of these group types and ranges. For example, the term "C _1-6 alkyl" refers particularly to the disclosure independently methyl, ethyl, C ₃ alkyl, C ₄ alkyl, C ₅ alkyl, and C ₆ alkyl.

Unless explicitly pointed out in other ways, the description methods used in the present invention are interchangeable with "... each independently being" and "... independently being", and should be understood in a broad sense. It means that in different groups, the specific options expressed between the same symbols do not affect each other. Taking the compound represented by formula I as an example, when n is 2, the specific options of the two Rs are not affected by each other.

As described in the present invention, the ring system (such as the compound shown in formula I) formed by attaching a substituent to the central ring by drawing a bond represents that the substituent can be substituted at any substitutable position on the ring. For example, the compound represented by formula I represents that the substituent R can be mono-substituted or multi-substituted at any position on the ring directly connected to the substituent that may be substituted, as shown in formula 1-7.

The term "prodrug" as used in the present invention represents the conversion of a compound into a compound represented by formula (I) in vivo. Such conversion is affected by the hydrolysis of the prodrug in the blood or the enzymatic conversion of the prodrug into the maternal structure in the blood or tissue. The prodrug compounds of the present invention may be esters. In the existing invention, esters may be used as prodrugs including phenyl esters, aliphatic (C ₁ -C ₂₄ ) esters, acyloxymethyl esters, and carbonic acid. Esters, carbamates and amino acid esters. For example, a compound in the present invention contains a hydroxyl group, which can be acylated to obtain a compound in the form of a prodrug. Other prodrug forms include phosphate esters. For example, these phosphate ester compounds are obtained by phosphorylation of the parent hydroxyl group. For a complete discussion of prodrugs, you can refer to the following literature: T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the ACSSymposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, J. Rautio et al., Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270, and SJ Hecker et al., Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry , 2008, 51,2328-2345.

"Metabolite" refers to the product obtained by the metabolism of a specific compound or its salt in the body. The metabolites of a compound can be identified by techniques well known in the art, and its activity can be characterized by experimental methods as described in the present invention. Such products can be obtained by oxidizing, reducing, hydrolyzing, acylating, deamidating, esterifying, degreasing, enzymatic cleavage and the like of the administered compound. Correspondingly, the present invention includes the metabolites of the compound, including the metabolites produced by fully contacting the compound of the present invention with a mammal for a period of time.

The "pharmaceutically acceptable salt" used in the present invention refers to the organic and inorganic salts of the compound of the present invention. Pharmaceutically acceptable salts are well-known in the field, as described in the literature: SMBerge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66:1-19. Pharmaceutically acceptable non-toxic acid salts include, but are not limited to, inorganic acid salts formed by reaction with amino groups include hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, And organic acid salts such as acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, or other methods described in books and literature such as ion exchange These salts. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphoric acid Salt, camphor sulfonate, cyclopentyl propionate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate Salt, gluconate, hemisulfate, heptanoate, caproate, hydroiodide, 2-hydroxy-ethanesulfonate, lacturonate, lactate, laurate, lauryl sulfate, Malate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionic acid Salt, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts obtained with appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. The present invention also contemplates the quaternary ammonium salt formed by any compound containing the N group. Water-soluble or oil-soluble or dispersed products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include appropriate, non-toxic ammonium, quaternary ammonium salts, and amine cations that resist counterion formation, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, and C _{1 -8} Sulfonates and aromatic sulfonates.

The "solvate" of the present invention refers to an association formed by one or more solvent molecules and the compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to the association formed by the solvent molecule being water.

When the solvent is water, the term "hydrate" may be used. In some embodiments, a compound molecule of the present invention may be combined with a water molecule, such as a monohydrate; in other embodiments, a compound molecule of the present invention may be combined with more than one water molecule, such as a dihydrate In some embodiments, one compound molecule of the present invention can be combined with less than one water molecule, such as a hemihydrate. It should be noted that the hydrate of the present invention retains the bioavailability of the compound in a non-hydrated form.

The term "treating" any disease or condition as used in the present invention refers to all that can slow, interrupt, prevent, control or stop the progression of the disease or condition, but does not necessarily mean that all symptoms of the disease or condition disappear, and it also includes Preventive treatment of the symptoms, especially in patients who are susceptible to such diseases or disorders. In some of these embodiments, it refers to ameliorating a disease or condition (ie, slowing down or preventing or reducing the development of the disease or at least one clinical symptom thereof). In other embodiments, "treatment" refers to alleviating or improving at least one physical parameter, including physical parameters that may not be perceived by the patient. In other embodiments, "treatment" refers to the regulation of the disease or condition physically (for example, stabilizing the perceptible symptoms) or physiologically (for example, stabilizing the parameters of the body) or both. In other embodiments, "treating" refers to preventing or delaying the onset, occurrence, or worsening of a disease or condition.

The term "composition" as used in the present invention refers to a product containing a prescribed amount of prescribed ingredients, and any product directly or indirectly produced by a combination of prescribed amounts of prescribed ingredients. The meaning of this term in relation to pharmaceutical compositions includes products containing active ingredients (single or multiple) and inert ingredients (single or multiple) constituting the carrier, and products composed of any two or more ingredients mixed, compounded or aggregated , Or any product produced directly or indirectly by the decomposition of one or more components, or by other types of reactions or interactions of one or more components. Therefore, the pharmaceutical composition of the present invention includes any composition prepared by mixing the compound of the present invention with a pharmaceutically acceptable carrier.

The invention provides a pharmaceutical composition for preventing and treating infertility or fertility difficulties caused by random mutations of mtDNA. The pharmaceutical composition comprises: the compound represented by formula I and its precursors and derivatives such as NADH or NAD ⁺ or a pharmaceutically acceptable salt thereof as active ingredients; and pharmaceutically acceptable excipients. The medicine is used to prevent and treat female infertility or improve female fertility.

In some embodiments, the pharmaceutical composition is in a dosage form of at least one of tablets, capsules, granules or injections.

In some embodiments, the pharmaceutical composition further comprises auxiliary substances, optionally, the auxiliary substances are wetting agents, emulsifiers, preservatives or buffers.

In some embodiments, the sterility or fertility difficulties caused by the random mutation of mtDNA is at least one selected from the group consisting of: elderly women who have accumulated random mutations of mtDNA, and female infertility caused by mitochondrial dysfunction caused by random mutations of mtDNA. . Among them, the "random mutation of mtDNA" in the "random mutation of mtDNA leading to mitochondrial dysfunction and thus female infertility" is caused by non-age factors, such as environmental factors.

The present invention will be further explained below in conjunction with specific embodiments.

The reagents and materials used in the following examples are all commercially available. If not clearly stated, the methods and conditions used are also processed in accordance with known methods and conditions.

Example 1 Detection of fertility in POLG mutant mice

Mitochondrial DNA polymerase (POLG) plays a very important role in the replication and repair of mitochondrial DNA. The loss of POLG function leads to mitochondrial dysfunction, which will cause the balance of ROS production and clearance to be broken, which may affect the function of adult stem cells. , The tissues and organs cannot be renewed and repaired, which makes the body appear aging. POLG mutant mice have a D257A mutation in the POLG exonuclease position, which causes the exonuclease activity to be lost, leading to correction defects in the mtDNA replication process and causing mtDNA mutations. The mice will accumulate a large amount of randomness during development. Mutations: including point mutations and deletion mutations, and have developed a variety of characteristics of premature aging. It is a good model for studying mitochondrial DNA mutations. The inventors selected 6 pairs of 2-month-old POLG mutant homozygous female and male mice (Mut), and they were caged separately to test the fertility of each pair of mice; at the same time, 6 pairs of 2-month-old wild-type female and male mice were selected. Male mice (WT) were caged together, and the fertility of each pair of mice was tested as a control. The inventor found that the fertility of the POLG mutant mice is very low, the average number of litters per pair of mice is only 1±0.4 in the first litter, and 0 in the second and third litters. The average number of litters per pair of wild-type mice in the first litter is 9.3±0.2, the second litter is 9.6±0.3, and the third litter is 8.5±0.2, as shown in Figure 1. These results indicate that mtDNA mutations can significantly reduce the fertility of mice, and make the fertility of mice basically lost.

Example 2 NMN improves the fertility of POLG mutant mice

Next, the inventor used the POLG mutant mouse as a model, and the inventor conducted a drug screening that can improve the fertility of mice. In the end, the inventors added 6-week-old POLG mutant female mice to 900 mg/kg/day drinking water with NMN, and water without NMN was used as a control. And after adding NMN or control water to raise the mice for 2 weeks, that is, after the mice were crossed with wild male mice at 8 weeks, the fertility of POLG mutant female mice was tested. The results are shown in Figure 2. The number of litters in the first litter of POLG mutant females treated with NMN (n=6) is greater than that of POLG mutant female mice treated with water (n=6). The results show that NMN can significantly improve the fertility of POLG mutant female mice.

In the description of this specification, descriptions with reference to the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" etc. mean specific features described in conjunction with the embodiment or example , Structure, materials or features are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the described specific features, structures, materials or characteristics can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine the different embodiments or examples and the characteristics of the different embodiments or examples described in this specification without contradicting each other.

Although the embodiments of the present invention have been shown and described above, it can be understood that the above-mentioned embodiments are exemplary and should not be construed as limiting the present invention. Those of ordinary skill in the art can comment on the foregoing within the scope of the present invention. The embodiment undergoes changes, modifications, substitutions and modifications.

Claims

The use of the compound represented by formula I or the stereoisomers, nitrogen oxides, solvates, metabolites, pharmaceutically acceptable salts, prodrugs or derivatives thereof in the preparation of medicines, Said drugs are used to treat or prevent female infertility,

among them:

Each R is independently H, D, F, Cl, Br, I, OH, NH 2 , NO 2 , CN, N 3 or C 1-6 alkyl;

n is 0, 1, 2, 3, or 4.
The use according to claim 1, characterized in that each R is independently H, D, F, Cl, Br, I, OH, NH 2 , NO 2 , CN, N 3 or C 1-4 alkyl;

Optionally, each R is independently H, D, F, Cl, Br, I, OH, NH 2 , NO 2 , CN, N 3 , methyl, ethyl, n-propyl, isopropyl, n-butyl Base, isobutyl or tert-butyl;

Optionally, the compound has the following structure:

Optionally, the derivative of the compound is NADH or NAD + .
The use according to claim 1, wherein the female infertility is caused by mtDNA mutations in egg cells;

Optionally, the female infertility is caused by a point mutation in mtDNA of the egg cell.
Use of the compound represented by formula I or the stereoisomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt, prodrug or derivative thereof of the compound represented by formula I in the preparation of a kit, The kit is used to restore mtDNA mutations,

among them:

Each R is independently H, D, F, Cl, Br, I, OH, NH 2 , NO 2 , CN, N 3 or C 1-6 alkyl;

n is 0, 1, 2, 3, or 4.
The use according to claim 4, characterized in that each R is independently H, D, F, Cl, Br, I, OH, NH 2 , NO 2 , CN, N 3 or C 1-4 alkyl;

Optionally, each R is independently H, D, F, Cl, Br, I, OH, NH 2 , NO 2 , CN, N 3 , methyl, ethyl, n-propyl, isopropyl, n-butyl Base, isobutyl or tert-butyl;

Optionally, the compound has the following structure:

Optionally, the derivative of the compound is NADH or NAD + .
Use of the composition in the preparation of a pharmaceutical composition for the treatment or prevention of female infertility, the composition comprising the compound defined in any one of claims 1 to 2.
The use according to claim 6, wherein the composition further comprises: a pharmaceutically acceptable excipient, carrier, adjuvant, vehicle, or a combination thereof.
The use according to claim 6, characterized in that the composition further comprises: other drugs for treating or preventing female infertility.
Use of the composition in the preparation of health care products for the prevention of female infertility, and the composition includes the compound defined in any one of claims 1 to 2.