Use of mevalonate in the manufacture of drugs that improve early embryonic development of senescent oocytes

Abstract

The present invention provides a use of mevalonic acid in the manufacture of a drug for improving early embryonic development of aged oocytes, which belongs to the field of biomedicine technology. Through experimental research, the present invention has found that mevalonic acid can be used to improve the fertilization rate, two-cell formation rate and good-quality blastocyst formation rate of aged oocytes, thereby improving the development process of early embryos, providing a new idea and method for infertility treatment.

Description

Technical Field

[0001] The present invention belongs to the field of biopharmaceutical technology, and specifically relates to the use of mevalonic acid in the manufacture of drugs for improving early embryo development of aged oocytes.

Background Art

[0002] The medical definition of infertility is the failure to achieve pregnancy despite having normal sexual intercourse without any contraceptive measures for over one year, and it is mainly divided into primary infertility and secondary infertility. Primary infertility means never having been pregnant, and secondary infertility means becoming infertile after having been pregnant once. Infertility is a common problem and is said to affect at least 10 - 15% of couples of childbearing age. The main causes of infertility are, in order, ovulation disorders, semen abnormalities, fallopian tube abnormalities, unexplained infertility, endometriosis, and other immunological infertility. Other factors include cervical factors such as cervical stenosis, which account for more than 5% of all cervical factors. The main causes of female infertility are ovulation disorders, fallopian tube factors, and endometrial receptivity abnormalities. The main causes of male infertility are spermatogenesis abnormalities and ejaculatory disorders. Compared with young women, women over 35 years old have a decline in the quality of oocytes, and the risks of infertility, miscarriage, and stillbirth increase. Especially women over 40 years old have a clinical pregnancy rate of less than 20% in in vitro fertilization (IVF). The decline in the quality of oocytes, fertilization rate, and good-quality blastocyst formation rate in older women are the main causes of the decline in pregnancy rate and the increase in miscarriage. Currently, there is a lack of effective means to improve the fertilization rate and good-quality blastocyst formation rate of oocytes in older patients.

[0003] The mevalonate pathway is a metabolic pathway that synthesizes cholesterol and isoprenoids using acetyl-A as a raw material, and plays a crucial role in human health and disease. Cholesterol itself not only controls cell membrane fluidity and regulates gene transcription, but can also synthesize steroid hormones as a precursor molecule. The mevalonate pathway also contains a series of important metabolic intermediates, such as farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP), which modify proteins through isoprenemination and exert important regulatory effects on cellular function. Furthermore, recent research has shown that mevalonic acid (MVA), an important metabolic intermediate of the mevalonate pathway, not only acts as a precursor for the synthesis of cholesterol and isoprenoids, but also activates the IGF1-R and mTOR signaling pathways and performs epigenetic modifications via histone function. Patent CN202110273233.9 discloses the use of geranylgeraniol (GGOH) or farnesol (FOH) in improving the quality and meiotic progression of aging ovarian oocytes. While this can improve the meiotic progression of aging oocytes, GGOH needs to be dissolved in an organic solvent, which may increase the risk when added to in vitro culture media.

[0004] Mevalonic acid supplementation can improve intracellular cholesterol content, increase protein isoprene modification, and enhance epigenetic modifications; however, there have been no reports on the effects of mevalonic acid on the fertilization or embryonic development processes of aging oocytes. [Overview of the Initiative] [Problems that the invention aims to solve]

[0005] To address the aforementioned shortcomings, the present invention provides the use of mevalonic acid (MVA) in the manufacture of a drug that improves early embryonic development of senescent oocytes. Through experiments, the present invention has discovered that mevalonic acid improves the fertilization rate, bicellular formation rate, and high-quality blastocyst formation rate of senescent oocytes, thereby improving the developmental process of early embryos and proposing a new approach and method in infertility treatment. [Means for solving the problem]

[0006] To achieve the objectives of the above invention, the following technical solutions of the present invention are proposed.

[0007] According to one embodiment, the present invention provides the use of mevalonic acid in the manufacture of a drug that improves the early embryonic development of senescent oocytes.

[0008] Specifically, the chemical formula of the aforementioned mevalonic acid is: As shown in JPEG0007858073000001.jpg3159.

[0009] In another aspect, the present invention provides the use of mevalonic acid in the manufacture of a drug that improves the fertilization rate, bicellular formation rate, and / or high-quality blastocyst formation rate of senescent oocytes.

[0010] Specifically, the chemical formula of the aforementioned mevalonic acid is: As shown in JPEG0007858073000002.jpg3159.

[0011] In another embodiment, the present invention provides a drug that improves the early embryonic development of senescent oocytes, the drug comprising mevalonic acid.

[0012] Specifically, the chemical formula of the aforementioned mevalonic acid is: As shown in JPEG0007858073000003.jpg3159.

[0013] Specifically, the concentration of the mevalonic acid is 10 to 100 μmol / L, preferably 50 μmol / L.

[0014] Specifically, the drug further comprises an optional, pharmaceutically acceptable carrier.

[0015] More specifically, the pharmaceutically acceptable carriers include, but are not limited to, diluents, excipients, fillers, wetting agents, disintegrants, flavoring agents, and adhesives. [Effects of the Invention]

[0016] Compared to conventional technologies, the positive and beneficial effects of the present invention are as follows:

[0017] 1. Through experimental research, the present invention has discovered that mevalonic acid can be used to improve the developmental process of early embryos by improving the fertilization rate, bicellular formation rate, and high-quality blastocyst formation rate of aging oocytes, thereby providing a new concept and method for infertility treatment. [Brief explanation of the drawing]

[0018] [Figure 1] This figure shows the results of fertilization medium supplemented with MVA, which improves the rate of bicellular formation in senescent oocytes. [Figure 2] This figure shows the results of embryonic medium supplemented with MVA, which improves the rate of high-quality blastocyst formation from senescent oocytes. [Figure 3] This is a diagram showing the safety evaluation results for offspring mice after MVA-treated blastocyst transfer. [Modes for carrying out the invention]

[0019] The present invention will be described in more detail below with reference to specific examples. The following examples are not intended to limit the present invention, but are used as examples to illustrate it. Unless otherwise specified, the experimental methods used in the following examples generally follow normal conditions, and unless otherwise specified, the materials, reagents, etc. used in the following examples are all commercially available.

[0020] Example 1. Improvement of fertilization rate, two-cell, and high-quality blastocyst formation rate of aged oocytes by mevalonic acid (MVA) 1) According to the following composition, a fertilization medium containing 50 μmol / L MVA and an embryo medium containing 50 μmol / L MVA were prepared under aseptic conditions.

[0021] Table 1 JPEG0007858073000004.jpg56156

[0022] Table 2 JPEG0007858073000005.jpg55156

[0023] 2) The epididymides of 8-week-old C57BL / 6 male mice were collected and transferred to a fertilization drop for acquiring fertilization ability (G-IVF Plus). Sperm were released with microsissors, and the fertilization ability was acquired for 1 hour.

[0024] 3) 10 IU of PMSG was injected intraperitoneally into 10-month-old C57BL / 6 female mice. After 48 h, 10 IU of HCG was injected intraperitoneally. After 13 h, ovaries and oviducts were separated from the M2 medium, and the ampulla of the oviduct was broken with microscopic forceps to release the cumulus mucous mass. The mucous mass was transferred to a fertilization medium containing 50 μmol / L MVA covered with mineral oil, sperm were added, and the culture was carried out in a 37 °C, 5% CO2 incubator.

[0025] 4) Four hours after fertilization in the fertilization medium containing 50 μmol / L MVA, sperm around the fertilized eggs were washed using a mouse pipette and then transferred to an embryo medium containing 50 μmol / L MVA. After 2 h, the fertilization rate was counted, and after 14 h, the two-cell formation rate was counted. The results were as shown in Figure 1.

[0026] 5) The development status of embryos was observed, and after 84 h of culture, the high-quality blastocyst formation rate was counted. The results were as shown in Figure 2.

[0027] 6) After selecting and transplanting high-quality blastocysts, a safety evaluation was performed. The results were as shown in Figure 3.

[0028] As can be seen from Figures 1 and 2, the addition of mevalonic acid (MVA) to the fertilization medium and embryo medium significantly improved the rate of two-cell formation and high-quality blastocyst formation from senescent oocytes compared to the control group. As can be seen from Figure 3, MVA treatment did not affect offspring safety. Therefore, MVA can be used in the production of drugs that improve the rate of two-cell formation from senescent oocytes and the rate of high-quality blastocyst formation.

[0029] The above-described embodiments merely represent several embodiments of the present invention, and while the descriptions are relatively specific and detailed, they should not be understood as limiting the scope of the patent for the present invention. It should be noted that, for those skilled in the art, some modifications and improvements are possible without departing from the concept of the present invention, and all of these fall within the scope of protection of the present invention. Therefore, the scope of patent protection for the present invention should be in accordance with the appended claims.

Claims

1. The use of mevalonic acid in the manufacture of drugs to improve early embryonic development of senescent oocytes, The chemical formula of the aforementioned mevalonic acid is, As shown, The mevalonic acid is used on the aging oocytes during and after fertilization. The use of mevalonic acid is characterized in that the drug is used to prepare a culture medium containing 50 μmol / L mevalonic acid.

2. The use of mevalonic acid in the manufacture of drugs for improving the fertilization rate, bicellular formation rate and / or high-quality blastocyst formation rate of senescent oocytes, The chemical formula of the aforementioned mevalonic acid is, As shown, The mevalonic acid is used on the aging oocytes during and after fertilization. The use of mevalonic acid is characterized in that the drug is used to prepare a culture medium containing 50 μmol / L mevalonic acid.

3. The use of mevalonic acid according to claim 1 or 2, characterized in that the drug further comprises an optional pharmaceutically acceptable carrier.

4. The use of mevalonic acid according to claim 3, characterized in that the pharmaceutically acceptable carrier includes, but is not limited to, diluents, excipients, fillers, wetting agents, disintegrants, flavoring agents, and adhesives.

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