Use of Furfurman in the preparation of an anti-radiation drug for the ovary or a drug for treating ovarian ionizing radiation injury

Abstract

The present invention relates to the use of Furfurman in the preparation of an anti-radiation drug for the ovary or a drug for treating ovarian ionizing radiation injury. A potential ovarian radiation protector, Furfurman, targeting the Dectin-2 receptor obtained in the present invention was used in animal model experiments. The results showed that it could improve the estrous cycle disorder in irradiated mice, increase the ovarian organ coefficient in irradiated mice, and improve the disorder of ovarian hormone levels in mice caused by irradiation, confirming that Furfurman can reduce radiation-induced ovarian tissue damage and bringing new hope for protecting against ovarian radiation injury.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and more particularly to the use of Furfurman in the preparation of drugs for anti-radiation of ovaries or for treating ovarian ionizing radiation injury. Background Art

[0002] At present, only a few young cancer patients receive genetic counseling before or during treatment. Fertility preservation has become the second most important issue to consider for this group of people after survival. "Oncofertility" has gradually developed in this context and has gradually become a relatively popular topic in recent years, attracting the attention of more and more clinical oncology doctors and fertility experts, and enabling more and more scholars to be committed to the research on fertility preservation measures for cancer patients.

[0003] In the specific practice of "Oncofertility", it mainly includes the following three aspects of measures: physical protection, biological protection, and drug protection. Physical protection mainly reduces the damage caused by radiation through a barrier effect, but it cannot effectively avoid the situation where, for example, cranial radiotherapy affects the hypothalamic-pituitary-gonadal axis, thereby leading to ovarian dysfunction, and the effect is not very satisfactory. In recent years, the development of ovarian transposition has provided a new option for the preservation of ovarian function in premenopausal female cancer patients who need pelvic radiotherapy, but it also faces some limitations, such as poor ovarian reserve capacity, high risk of ovarian metastasis, and only undergoing chemotherapy alone. Biological protection, including embryo cryopreservation and transplantation, oocyte cryopreservation and transplantation, ovarian tissue cryopreservation and transplantation, and ovarian transplantation, etc., is the most widely carried out project in the practice of fertility preservation for premenopausal female cancer patients at present. However, this type of protection method also faces many challenges and limitations. For example, the technical operation in the specific practice of this type of project is relatively difficult, there are few medical institutions that carry out such technologies, social and family economic factors, and related medical contraindications, etc. Drug protection measures have received the attention of many scholars due to their relatively simple operation process and little conflict with cancer treatment, and certain progress has been made.

[0004] In terms of drug protection, the currently best drug internationally is WR-2721 approved by the FDA. However, it has been proven to be highly toxic and have many side effects during practice, which to a certain extent limits its use. The currently available drugs for preventing and treating radiation damage in China include estrogen and Rubia cordifolia extract, etc., and their efficacy has been proven to be poor during practice. In recent years, Chinese scholars have carried out a number of basic research in the field of traditional Chinese medicine, and the results suggest that various traditional Chinese medicines or decoctions can reduce radiation-induced ovarian tissue damage, bringing new hope for reducing ovarian radiation damage protection. However, they all face multiple challenges such as unclear effects, unclear specific mechanisms, and relatively large toxic and side effects. Therefore, there is an urgent need to develop drugs with high efficiency and low toxicity that target new targets of ionizing radiation.

[0005] In recent years, it has been reported and confirmed that pattern recognition receptors (PRRs) play an important role in anti-radiation damage [1-3]. Therefore, we screened relevant PRRs and their ligands, and obtained a potential ovarian radiation protector Furfurman that targets the Dectin-2 receptor, which is mainly derived from the cell wall of Malassezia furfur. After Furfurman binds to the Dectin-2 receptor, Dectin-2 can be coupled with the Fc receptor gamma chain, and signal through the kinase Syk and the adaptor CARD9 / Bcl10 / MALT1, triggering the activation of the NF-κB pathway and the production of downstream related products, thereby exerting its effect.

[0006] At present, there is no report on the use of the Furfurman of the present invention in the preparation of drugs for ovarian anti-radiation or treating ovarian ionizing radiation damage. Summary of the Invention

[0007] The first object of the present invention is to provide a use of Furfurman in view of the deficiencies in the prior art.

[0008] The second object of the present invention is to provide a pharmaceutical composition.

[0009] To achieve the above first object, the technical solution adopted by the present invention is:

[0010] The application of Furfurman in the preparation of drugs for ovarian anti-radiation or treating ovarian ionizing radiation damage.

[0011] As a preferred example, the radiation is γ-ray or X-ray radiation.

[0012] More preferably, the Furfurman targets the Dectin-2 receptor, which is coupled to the Fc receptor gamma chain and signals through the kinase Syk and the adaptors CARD9 / Bcl10 / MALT1 to trigger the activation of the NF-κB pathway and the production of downstream related products to exert its effect.

[0013] To achieve the second above-mentioned object, the technical solution adopted by the present invention is:

[0014] A pharmaceutical composition for anti-radiation of the ovary or treatment of ovarian ionizing radiation injury, wherein the drug comprises Furfurman and one or more pharmaceutically acceptable carriers or excipients.

[0015] As a preferred example, the pharmaceutically acceptable carrier or excipient is selected from one or several of solvents, solubilizers, cosolvents, emulsifiers, flavoring agents, odor correctors, coloring agents, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, pH regulators, stabilizers, surfactants or preservatives.

[0016] More preferably, the dosage form is a solid preparation, a semi-solid preparation or a liquid preparation.

[0017] More preferably, the solid preparation includes tablets, capsules, granules and / or pills; the semi-solid preparation includes gels, suppositories and / or ointments; the liquid preparation includes emulsions, mixtures, suspensions and / or solutions.

[0018] The advantages of the present invention are:

[0019] It brings new hope for reducing the radiation injury protection of the ovary and can be used to develop drugs with high efficiency and low toxicity and targeting new targets of ionizing radiation. Description of the Drawings

[0020] Appendix Figure 1 Changes in the organ coefficient of the ovarian tissue of mice after irradiation (IR+NS: simple irradiation group;

[0021] IR+FFM: drug-administered irradiation group) (*: P < 0.05).

[0022] Appendix Figure 2Effect of Furfurman on the disorder of ovarian hormone levels in irradiated mice. (A) Protective effect of Furfurman on anti-Müllerian hormone (AMH) in mice; (B) Protective effect of Furfurman on estrogen (E2) in mice; (C) Protective effect of Furfurman on follicle-stimulating hormone (FSH) in mice; (D) Protective effect of Furfurman on luteinizing hormone (LH) in mice. (IR + NS: simple irradiation group; IR + FFM: drug-administered irradiation group) (**: P < 0.01, ***: P < 0.001).

[0023] Appendix Figure 3 Follicle counts of mice in each group. (A) Primordial follicle counts of mice in each group; (B) Primary follicle counts of mice in each group; (C) Preantral follicle counts of mice in each group; (D) Antral follicle counts of mice in each group; (E) Atretic follicle counts of mice in each group; (F) Total follicle counts of mice in each group. (IR + NS: simple irradiation group; IR + FFM: drug-administered irradiation group) (*: P < 0.05, **: P < 0.01, ***: P < 0.001). Specific implementation manners

[0024] The present invention will be further described below in conjunction with specific implementation manners. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. In addition, it should be understood that after reading the content described in the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

[0025] Example 1

[0026] 1. Experimental materials

[0027] Experimental animals: The genetic background of the mice selected in this experiment is C57BL / 6J, purchased from Shanghai Jihui Experimental Animal Breeding Co., Ltd., with a weekly age of 6 - 8 weeks, and raised in an animal room with day-night alternating lighting and a constant temperature of 20°C.

[0028] Main reagents: Furfurman (InvivoGen); ELISA detection reagents for mouse estrogen (E2), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and anti-Müllerian hormone (AMH): Shanghai Zhuocai Biotechnology Co., Ltd.; hematoxylin dye, eosin dye, Giemsa dye (Zhuhai Baisuo Biotechnology Co., Ltd.).

[0029] 2. Experimental methods:

[0030] (a) Animal model establishment: Female mice were divided into a simple irradiation group and a drug-administered irradiation group. All female mice in the drug-administered irradiation group were intraperitoneally injected with Furfurman (drug concentration: 50 mg / Kg) 12 hours and 2 hours before irradiation. Female mice in the simple irradiation group were intraperitoneally injected with the same volume of normal saline. All female mice received a single pelvic local radiotherapy of 2 Gy after the drug administration was completed.

[0031] (b) Experimental arrangement: 16 female mice were divided into two groups. After drug administration and irradiation, subsequent experiments such as organ coefficient measurement, follicle counting, estrous cycle determination, and ovarian hormone level detection were carried out.

[0032] (c) Estrous cycle: Starting 24 hours after the irradiation of the mice, at 08:00 and 20:00 every day, the morphological changes of vaginal exfoliated cells of each group of mice were detected to determine the specific stage of the estrous cycle of the mice. The entire 21 days were fully evaluated and statistical analysis was performed.

[0033] (d) Hormone levels: After the estrous cycle assessment experiment was completed, the mice were anesthetized, and blood was taken from the eyeballs to obtain serum. The levels of AMH, E2, FSH, and LH in the mice were detected by ELISA method.

[0034] (e) Organ coefficient: After the anesthesia ended, the weights of each group of mice were measured. After the blood was taken from the eyeballs, the bilateral ovarian tissues of the mice were dissected and weighed, and the organ coefficient was calculated and analyzed.

[0035] (f) Follicle counting: The ovarian tissues of the mice were used to make paraffin specimens, and the specimens were subjected to follicle counting to evaluate the changes in the number of follicles at all levels and the total number of follicles.

[0036] 3. Statistical analysis

[0037] Analysis was performed using GraphPad Prism 9.0 and SPSS 22.0 software. All data were expressed as mean ± standard deviation (Means ± SD). Unpaired t-tests were used for the differential analysis between measurement data. P < 0.05 was considered to be statistically significantly different.

[0038] 4. Experimental results:

[0039] 4.1. Furfurman administration can improve the estrous cycle disorder in irradiated mice

[0040] During the three batches of experiments, some mice in both the irradiation group and the irradiation + drug administration group showed estrous cycle disorders. Compared with the simple irradiation group, 72.22 ± 9.62% of the mice in the simple irradiation group had estrous cycle disorders, and the estrous cycle disorder rate in the irradiation + drug administration group was 55.56 ± 9.62%. There was a statistical difference between the two groups, as shown in Table 1.

[0041] Table 1 Estrus cycle disorder rate of mice in each group

[0042]

[0043] Note: (IR + NS: simple irradiation group; IR + FFM: drug-administered irradiation group) (*: P < 0.05)

[0044] 4.2 Furfurman can increase the ovarian organ coefficient of irradiated mice

[0045] In terms of the ovarian organ coefficient, compared with the control group, the ovarian organ coefficient of mice in the simple irradiation group was significantly reduced. Intraperitoneal injection of Furfurman to mice before irradiation would reverse this trend to a certain extent. There was a significant difference between the two groups statistically compared with the simple irradiation group, as shown in Figure 1 .

[0046] 4.3 Furfurman can effectively improve the disorder of ovarian hormone levels in irradiated mice

[0047] Compared with the simple irradiation group, application of Furfurman to mice before irradiation would significantly increase the levels of anti-Müllerian hormone ( Figure 2 A) and estrogen ( Figure 2 B) in irradiated mice. At the same time, it could significantly reduce the levels of follicle-stimulating hormone ( Figure 2 C) and luteinizing hormone ( Figure 2 D) in irradiated mice, as shown in Figure 2 .

[0048] 4.4 Effects of Furfurman on follicles in irradiated mice

[0049] Furfurman can increase the number of follicles at all levels of effective follicles ( Figure 3 A / B / C / D), reduce the number of atretic follicles ( Figure 3 E), and increase the total number of follicles ( Figure 3 F). There was a statistical difference between the two, as shown in Figure 3 .

[0050] References

[0051] [1] GAO F, ZHANG C, ZHOU C, et al. A critical role of toll-like receptor 2 (TLR2) and its 'in vivo ligands in radio-resistance[J]. Sci Rep, 2015, 5(13004.

[0052] [2]LIU C, ZHANG C, MITCHEL R E, et al. A critical role of toll-like receptor 4(TLR4) and its 'in vivo ligands in basal radio-resistance[J]. Cell death & disease, 2013, 4(e649.

[0053] [3]LIU L, QU H, QIN H, et al. NOD2 agonist murabutide alleviates radiation-induced injury through DNA damage response pathway mediated by ATR[J]. Journal of cellular physiology, 2019, 234(11): 21294 - 306.

[0054] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the method of the present invention, several improvements and supplements can be made, and these improvements and supplements should also be regarded as the protection scope of the present invention.

Claims

1. Use of Furfurman in the preparation of an anti-radiation drug for the ovary or a drug for treating ovarian ionizing radiation injury.

2. The application according to claim 1, wherein The radiation is γ-ray or X-ray radiation.

3. The application according to claim 1, wherein The Furfurman targets the Dectin-2 receptor, and the Dectin-2 is coupled with the Fc receptor gamma chain and signals through the kinases Syk and the adaptors CARD9 / Bcl10 / MALT1 to trigger the activation of the NF-κB pathway and the production of downstream related products to exert effects.

4. Use of a pharmaceutical composition for anti-radiation of ovary or treating ovarian ionizing radiation injury in preparing an anti-radiation drug for ovary or a drug for treating ovarian ionizing radiation injury, characterized in that, The drug includes Furfurman and one or more pharmaceutically acceptable carriers or excipients.

5. The application according to claim 4, characterized in that The pharmaceutically acceptable carrier or excipient is selected from one or several of solvents, solubilizers, cosolvents, emulsifiers, flavoring agents, odor-correcting agents, coloring agents, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, pH regulators, stabilizers, surfactants or preservatives.

6. The application according to claim 5, characterized in that, The dosage form of the drug is a solid preparation, semi-solid preparation or liquid preparation.

7. The application according to claim 6, wherein The solid preparation includes tablets, capsules, granules and / or pills; the semi-solid preparation includes gels, suppositories and / or ointments; the liquid preparation includes emulsions, mixtures, suspensions and / or solutions.

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