Application of indirubin in preparation of medicine for treating oligospermia and asthenospermia

Abstract

The invention relates to the technical field of biological medicines, in particular to application of indirubin in preparation of a medicine for treating oligoasthenospermia. The indirubin disclosed by the invention is a natural product or an artificially synthesized product, is high in safety and small in toxic and side effects, and has a good effect of treating oligoasthenospermia.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to the application of indirubin in the preparation of drugs for treating oligospermia and asthenospermia. Background Technology

[0002] Indirubin (IND), an indigo dimer derivative, is also extracted from the traditional Chinese medicine Qingdai, which is made from the fermented leaves of the plant *Indigofera tinctoria*. Indirubin possesses various biological functions, including hemostatic, antipyretic, anti-inflammatory, antibacterial, and antiviral properties. Currently, indirubin has undergone extensive pharmacological research and is primarily used in the clinical treatment of chronic myeloid leukemia, exhibiting advantages such as low toxicity and few side effects. Indirubin is currently a high-value raw material for drug development and a strictly prescribed drug ingredient. Oral administration trials in mice have confirmed its high safety profile. Due to its numerous highly effective physiological activities and high safety, indirubin can be used as a therapeutic component for oligospermia and asthenospermia. Our previous studies have shown that indirubin can improve male reproductive system diseases.

[0003] However, there are still challenges in the treatment of oligospermia and asthenospermia. Existing Western medicines have single targets and limited efficacy. Natural substances derived from traditional Chinese medicine are edible and non-toxic, making the development of drugs for the treatment of oligospermia and asthenospermia of such natural products of great significance. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide an application of indirubin in the preparation of a drug for treating oligoasthenospermia. Indirubin is a natural product or a synthetic product, which has high safety, low toxicity and side effects, and has a good therapeutic effect on oligoasthenospermia.

[0005] To solve the above-mentioned technical problems, embodiments of the present invention provide an indigo red, the molecular formula of which is shown in Formula I:

[0006] Formula I,

[0007] The indigo red is a purple powder, mp: 356-358℃; EI-MS m / Z: 263.261.

[0008] This invention also provides the application of indirubin in the preparation of drugs for treating oligospermia and asthenospermia.

[0009] Preferably, the indirubin is used to prepare a drug for treating oligospermia and asthenospermia, with an effective dose ranging from 25 mg / mL to 100 mg / mL.

[0010] Preferably, the indirubin used as a drug for treating oligoasthenospermia is in the form of oral preparations, capsules, granules, powders, pills, targeted preparations, sustained-release preparations, controlled-release preparations, injections, or other routes of administration.

[0011] The beneficial effects of the above technical solution of the present invention are as follows:

[0012] The present invention relates to the application of indirubin in the preparation of drugs for treating oligoasthenospermia. Indirubin is a natural or synthetic product with high safety, low toxicity and side effects, and has a good therapeutic effect on oligoasthenospermia. Attached Figure Description

[0013] Figure 1 The hydrogen spectrum of indirubin;

[0014] Figure 2 The carbon spectrum of indigo red;

[0015] Figure 3 HE staining images of testicular tissue from the normal group, model group, and low, medium, and high concentration groups of indirubin in Example 2;

[0016] Figure 4 The images show the epididymal sperm content staining in the normal group, model group, and low, medium, and high concentration groups of indirubin in Example 2.

[0017] Figure 5 These are images of the testes in the normal group, model group, and low, medium, and high concentration groups of indigo red in Example 2.

[0018] Figure 6 HE staining images of testicular tissue from the normal group, oligoasthenospermia group, and indirubin treatment group in Example 3;

[0019] Figure 7 The image shows the sperm count results for the normal group, the oligoasthenospermia group, and the indirubin treatment group in Example 3. Detailed Implementation

[0020] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with the accompanying drawings and specific embodiments.

[0021] An embodiment of the present invention provides an indigo red, the molecular formula of which is shown in Formula I:

[0022] Formula I,

[0023] The indigo red is a purple powder, mp: 356-358℃; EI-MS m / Z: 263.261.

[0024] This invention also provides the use of indirubin in the preparation of a drug for treating oligoasthenospermia. The indirubin is used to prepare the drug for treating oligoasthenospermia, with an effective dose ranging from 25 mg / mL to 100 mg / mL.

[0025] The indirubin used as a drug for treating oligoasthenospermia is available in oral formulations, capsules, granules, powders, pills, targeted formulations, sustained-release formulations, controlled-release formulations, injections, and other routes of administration.

[0026] The technical solution of the present invention will be further described below with reference to specific embodiments.

[0027] Example 1: Preparation of Indigo Red

[0028] Indorubicin (0.01 mol) was dissolved in 50 mL of methanol and placed in a 150 mL round-bottom flask equipped with a reflux condenser and a stir bar. After reflux, KBH4 (0.27 g, 0.005 mol) was gradually added. The reaction mixture was stirred under reflux for 1 hour. During heating, the stopper of the round-bottom flask was removed three times, and the flask was shaken to ensure full contact between the reaction mixture and air. After 1 hour, the solution was cooled to room temperature, and indorubicin precipitated from the solution. The solid was separated by filtration and recrystallized from 95% ethanol to give 1.1 g (84%) of a purple-red solid. The preparation reaction formula of indorubicin is as follows:

[0029] .

[0030] Figure 1 This is the hydrogen spectrum of indigo red. 1 H NMR (600 MHz, DMSO-d6): δ 11.02 (s, 1H), 10.92 (s, 1H), 8.77 (d, J =7.8 Hz, 1H), 7.65 (d, J = 7.2 Hz, 1H), 7.58 (t, J = 4.8Hz, 1H), 7.42 (d, J =8.4 Hz, 1H), 7.26 (t, J =7.8 Hz, 1H), 7.02 (t, J =7.2 Hz, 2H), 6.91 (d, J = 7.8 Hz, 1H).

[0031] Figure 2 This is the carbon spectrum of indigo red. 13 C NMR (150 MHz, DMSO-d6) d(ppm): 189.12, 171.42,152.99, 141.39, 138.82, 137.57, 129.74, 125.16, 124.83, 121.94, 121.74,119.51, 113.92, 110.06, 107.05.

[0032] Example 2: The effect of indigo carmine on improving oligospermia and asthenospermia

[0033] The experimental steps for using indigo red to improve oligospermia and asthenospermia are as follows:

[0034] Fifty healthy male ICR mice, 7 weeks old, were housed in standard polycarbonate cages with corn cob padding, under standard environmental conditions (12:12h light / dark cycle, temperature 24°C, relative humidity 55%). Standard rodent feed and water were provided freely.

[0035] Before the experiment, all mice were acclimatized to the new animal facility for at least one week. Subsequently, the mice were randomly divided into five groups: a normal group, an oligoasthenospermia group (model group), and low, medium, and high concentration groups of indirubin (25 mg / kg, 50 mg / kg, and 100 mg / kg), with 10 mice in each group. Except for the normal group, all other groups were administered 50 mg / kg bw bisphenol A (BPA) to induce the model (BPA solution was prepared using corn oil as a solvent). After 28 days, the BPA-induced oligoasthenospermia model was successfully established.

[0036] Subsequently, the control group was administered an equal volume of corn oil by gavage, while the treatment group received indirubin at concentrations of 25, 50, and 100 mg / kg bw. Treatment was administered by gavage for 28 consecutive days. After the experiment, the patients were anesthetized and euthanized, and testicular sections were stained with hematoxylin and eosin to observe the drug efficacy.

[0037] Figure 3 , Figure 4 , Figure 5 The results of the experiment on the promoting effect of indirubin on oligoasthenospermia in mice (high concentration of indirubin can significantly promote the recovery of sperm concentration and improve testicular tissue morphology in oligospermia model mice).

[0038] Figure 3 HE staining images of testicular tissue from the normal group, model group, and low, medium, and high concentrations (25 mg / kg, 50 mg / kg, and 100 mg / kg) of indirubin. Figure 3 The results showed that the testicular tissue in the model group exhibited significantly more vacuolated characteristics compared to the normal group. Low, medium, and high concentrations of indirubin (25 mg / kg, 50 mg / kg, and 100 mg / kg) all showed concentration-dependent improvement in oligoasthenospermia lesions in mice, with the high concentration (100 mg / kg) group showing the best efficacy.

[0039] Figure 4 The images show the epididymal sperm content in the normal group, model group, and low, medium, and high concentrations (25 mg / kg, 50 mg / kg, and 100 mg / kg) of indirubin. Figure 4 The results showed that the epididymal sperm count in the model group was significantly lower than that in the normal group, while the high concentration (100 mg / kg) of indirubin significantly increased the epididymal sperm count after administration.

[0040] Figure 5 The images show the appearance of the testes in the normal group, the model group, and the low, medium, and high concentration (25 mg / kg, 50 mg / kg, and 100 mg / kg) groups of indirubin. Figure 5 The results showed that the testes in the model group were significantly smaller than those in the normal group. The high concentration of indirubin (100 mg / kg) significantly improved the appearance of the testes and restored them to normal.

[0041] Example 3: The effect of indirubin in improving oligospermia and asthenospermia caused by chemotherapy drugs (busulfan)

[0042] The experimental steps for using indirubin to improve oligospermia and asthenospermia induced by chemotherapy drugs (busulfan) are as follows:

[0043] Fifty healthy male ICR mice, 7 weeks old, were housed in standard polycarbonate cages with corncob bedding, five mice per cage, under standard environmental conditions (12:12h light / dark cycle, temperature 24°C, relative humidity 55%). Standard rodent feed and water were provided freely. All mice were acclimatized to the new animal facility for at least one week before the experiment. Subsequently, the mice were randomly divided into 5 groups: a normal control group, an oligoasthenospermia group, and a 100 mg / kg indirubin treatment group, with 10 mice in each group. Except for the normal control group, all other groups were given a single intraperitoneal injection of 30 mg / kg bw busulfan (busulfan solution prepared using an aqueous solution containing 5% dimethyl sulfoxide as the solvent) to induce the model.

[0044] Fourteen days later, a busulfan-induced oligoasthenospermia model was successfully established. Subsequently, the normal control group was administered an equal volume of distilled water by gavage, while the indirubin treatment group received indirubin at a concentration of 100 mg / kg bw. Treatment was administered by gavage for 28 consecutive days. After the experiment, the animals were anesthetized and euthanized, and testicular sections were stained with hematoxylin and eosin to observe the drug efficacy.

[0045] Figure 6 HE staining images of testicular tissue from the normal group, the oligoasthenospermia group, and the indirubin treatment group. Figure 6 The results showed that, compared with the normal group, the testicular tissue morphology of the oligoasthenospermia group was significantly damaged, the spermatogenesis process was disordered, and spermatogenic cells were lost; compared with the oligoasthenospermia group, the testicular tissue morphology, spermatogenesis process, and spermatogenic cells of mice in the indirubin-treated group were significantly restored.

[0046] Figure 7 The graph shows the sperm count results for the normal group, the oligoasthenospermia group, and the indirubin treatment group. Figure 7 The results showed that compared with the normal group and the oligoasthenospermia group, the sperm count in the oligoasthenospermia group was significantly lower (P<0.05), while the sperm count in the indirubin treatment group was significantly higher than that in the oligoasthenospermia group, and the difference was statistically significant (P<0.05).

[0047] The above experiments show that indirubin has a significant therapeutic effect on the oligoasthenospermia model induced by busulfan in mice. After continuous administration for 4 weeks, the sperm count of oligospermia mice was significantly increased, and the testicular lesions and appearance were significantly improved. Indirubin has a concentration-dependent improvement effect on oligoasthenospermia mice induced by busulfan.

[0048] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An indigo red, characterized in that, Its molecular formula is as shown in Formula I: Equation I.

2. The indigo red according to claim 1, characterized in that, The indigo red is a purple powder, mp: 356-358℃; EI-MS m / Z: 263.

261.

3. The use of indirubin as described in any one of claims 1 to 2 in the preparation of a drug for treating oligoasthenospermia.

4. The application according to claim 3, characterized in that, The indirubin is used to prepare a drug for treating oligospermia and asthenospermia, with an effective dose ranging from 25 mg / mL to 100 mg / mL.

5. The application according to claim 3, characterized in that, The indirubin used as a drug for treating oligoasthenospermia is available in oral formulations, capsules, granules, powders, pills, targeted formulations, sustained-release formulations, controlled-release formulations, injections, and other routes of administration.

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