Application of FGF10 (fibroblast growth factor 10) in preparation of medicine for treating hyponatremia

Abstract

The invention discloses an application of FGF10 (fibroblast growth factor 10) in preparation of a medicine for treating hyponatremia, and experimental results show that the FGF10 is used for reducing the concentration of septidin in serum so as to increase the concentration of blood sodium in the serum and can be used for treating the hyponatremia.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to an application of FGF10 (fibroblast growth factor 10) in preparing a medicine for treating hyponatremia. Background Art

[0002] Hyponatremia (serum sodium <135mmol / L) is one of the most common electrolyte disorders and often occurs as a symptom of other diseases. Its prevalence is as high as 30% in acute and chronic hospitalized patients, and even higher in intensive care units. Studies have shown that hyponatremia is associated with patient readmission rates, short-term adverse events, and poor prognosis. In addition, hyponatremia is a recognized predictor of patient mortality. The mortality rate of patients with severe hyponatremia (serum sodium <120mmol / L) is as high as 20% (Adrogué HJ, et al. N Engl J Med, 2000, 342(21):1581-9; Adrogué HJ, et al. JAMA, 2022, 328(3):280-291.).

[0003] The most common cause of hyponatremia is the syndrome of inappropriate antidiuretic hormone secretion (SIAD). SIAD refers to the irregular release of arginine vasopressin (AVP) by the central nervous system under non-osmotic stimulation, which binds to AVPR2 to regulate glomerular filtration rate and sodium metabolism, ultimately leading to water retention and decreased blood sodium (Warren AM, et al. Endocr Rev, 2023, 44(5):819-861; Adrogué HJ, et al. N Engl J Med. 2023, 389(16):1499-1509.).

[0004] Currently, the main treatment options for patients with SIAD-related hyponatremia include water restriction, sodium supplementation, selective AVP type II receptor (V2) antagonists, urea, furosemide, etc. However, patients often find it difficult to tolerate the symptoms of thirst during water restriction treatment, resulting in poor compliance and cooperation. V2 receptor antagonists are not organ-specific and can cause certain adverse reactions to patients, such as hyperglycemia. A new and effective strategy to regulate AVP is urgently needed to further alleviate the adverse consequences of hyponatremia (Spasovski G, et al. Intensive Care Med, 2014, 40(3):320-31; Gross PA, et al. Kidney Int, 2011, 80(6):594-600; Ellison DH, et al. NEngl J Med, 2007, 356(20):2064-72.).

[0005] Previous literature reports clearly show the relationship between abnormal increase in AVP and hyponatremia (Berl TN Engl J Med, 2015, 372(23):2207-16; Hoorn EJ, et al. J Am Soc Nephrol, 2017, 28(5):1340-1349.). However, it is difficult to construct a SIAD model. Therefore, the present invention uses a bilateral adrenalectomy model to simulate abnormal AVP secretion in vivo to further intervene in hyponatremia caused by abnormal AVP secretion (Yamagata S, et al. Endocrinology, 2023, 164(8):bqad109.). Because AVP has a half-life of only 3 minutes in the blood and is difficult to detect, the present invention uses AVP precursor copeptin to replace the serum level of AVP for detection (Christ-Crain M, et al. Nat Rev Endocrinol, 2016, 12(3):168-76).

[0006] Fibroblast growth factor 10 (FGF-10) is a heparin-binding growth factor that belongs to the FGF family. Proteins in this family play a central role in the prenatal development, postnatal growth, and regeneration of various tissues by promoting cell proliferation and differentiation. Currently, there are no reports that FGF-10 has a therapeutic effect on hyponatremia. Summary of the Invention

[0007] In order to solve the above problems, the present invention provides a use of FGF10 in the preparation of a drug for treating hyponatremia, which can regulate hyponatremia caused by abnormal secretion of AVP.

[0008] The technical solutions of the present invention are as follows:

[0009] A use of FGF10 in preparing a medicine for treating hyponatremia.

[0010] Preferably, the hyponatremia is hyponatremia caused by excessive release of arginine vasopressin in the circulation system.

[0011] Preferably, the FGF10 is used to reduce the concentration of copeptin in serum, thereby increasing the blood sodium concentration in serum.

[0012] Preferably, the FGF10 is used to regulate the abnormal increase of arginine vasopressin.

[0013] Preferably, the drug for treating hyponatremia is a pharmaceutical preparation containing FGF10 or a pharmaceutical preparation containing an FGF10 promoter.

[0014] Compared with the prior art, the beneficial effects of the present invention are embodied in:

[0015] The present invention discovered for the first time that FGF10 has a clear therapeutic effect on hyponatremia caused by abnormal AVP secretion, and therefore can be applied to the treatment of AVP-related hyponatremia. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments.

[0017] Figure 1 In Example 1, bilateral adrenalectomy resulted in an abnormal increase in serum copeptin in mice, causing the mice to develop hyponatremia.

[0018] Figure 2 As in Example 2, when FGF10 was administered through the nasal cavity, the copeptin level of mice decreased and the blood sodium level increased.

[0019] Figure 3 In Example 3, there was no significant change in the body weight of mice after administration of FGF10.

[0020] Figure 4 In Example 4, FGF10 was administered to AVP-positive neurons, and the firing frequency of the neurons decreased. DETAILED DESCRIPTION

[0021] The present invention provides the use of FGF10 in the treatment of AVP-related hyponatremia. The active ingredient in the treatment of AVP-related hyponatremia described in the present invention is FGF10. The FGF10 used in this invention is provided by the Growth Factor Research Institute of Wenzhou Medical University, patent number: CN201910550314.1. This invention utilizes FGF10 to regulate AVP levels, thereby alleviating or treating hyponatremia. Furthermore, FGF10 has no significant toxic side effects upon administration, demonstrating excellent drug safety.

[0022] Example 1

[0023] Experimental process:

[0024] (1) Animal source: The experimental animals were 12 clean-grade male C57BL / 6J mice (7 weeks old, 20±4 g / mouse, purchased from Vital River Company).

[0025] Breeding environment requirements: good ventilation and air filtration system, quiet environment, room temperature maintained at 20℃, humidity controlled at 4-5%, ensure the litter is clean, and free access to water and food.

[0026] (2) Experimental groups: The mice were divided into two groups: a sham operation group and a bilateral adrenalectomy group (ADX), with 6 mice in each group.

[0027] (3) Model establishment:

[0028] ADX group: After the mice were anesthetized with isoflurane, a surgical incision was made parallel to the midline of the back to fully expose the adrenal glands. The adrenal glands and surrounding adipose tissue were removed with surgical forceps, and the surgical incisions were sutured layer by layer. Penicillin was administered daily after surgery to prevent infection.

[0029] Sham operation group: mice were not treated with any treatment after adrenal gland exposure, and the rest of the steps were the same as those of the ADX group.

[0030] (4) Serum collection: After surgery, mice in the sham and ADX groups recovered in clean cages for 3 days. They were lightly anesthetized with isoflurane, and 0.5 mL of blood was collected from the ophthalmic artery. The blood was kept at 4°C overnight and centrifuged at 2000 g for 15 minutes, and the supernatant was collected.

[0031] (5) Serum copeptin detection: A purchased ELISA kit (#LS-F7101, Lifespan, USA) was used for the detection of copeptin.

[0032] (6) Serum sodium test: Serum sodium was tested using an automatic biochemical analyzer (IDEXX, Catalyst One, USA).

[0033] Experimental results

[0034] like Figure 1 As shown in the figure, compared with the sham-operated mice, the level of copeptin in the serum of mice was significantly increased (P<0.05) and the level of blood sodium was significantly decreased (P<0.005) after bilateral adrenalectomy.

[0035] Figure 1 Middle: A is the serum copeptin level of mice in each group; B is the serum sodium level of mice in each group. **P<0.005, *P<0.05, n=12.

[0036] Example 2

[0037] Experimental process:

[0038] (1) Animal source: The experimental animals were 10 clean-grade male C57BL / 6J mice (7 weeks old, 20±4 g / mouse, purchased from Vital River Company).

[0039] Breeding environment requirements: good ventilation and air filtration system, quiet environment, room temperature maintained at 20℃, humidity controlled at 4-5%, ensure the litter is clean, and free access to water and food.

[0040] (2) Experimental grouping: The mice were divided into two groups: a control group and an FGF10-treated group, with 5 mice in each group.

[0041] (3) Model establishment:

[0042] FGF10-administered group: After mice were anesthetized with isoflurane, a surgical incision was made parallel to the midline of the back to fully expose the adrenal glands. The adrenal glands and surrounding adipose tissue were removed with surgical forceps, and the surgical incisions were sutured layer by layer. Penicillin was administered daily after surgery to prevent infection. On the fourth day after surgery, 10 μl of FGF10 solution (concentration: 1 mg / ml, solvent: sterile PBS, prepared using lyophilized powder of 201910550314.1) was administered through the nasal cavity once every 4 hours for three consecutive times.

[0043] Control group: mice were given 10 μl of PBS solution, and the rest of the steps were the same as those in the FGF10-administered group.

[0044] (4) Serum collection: After surgery and administration, mice in the control group and the FGF10-treated group were lightly anesthetized with isoflurane anesthesia. 0.5 mL of blood was collected from the ophthalmic artery, placed at 4°C overnight, and centrifuged at 2000 g for 15 minutes, and the supernatant was collected.

[0045] (5) Serum copeptin detection: A purchased ELISA kit (#LS-F7101, Lifespan, USA) was used for the detection of copeptin.

[0046] (6) Serum sodium test: Serum sodium was tested using an automatic biochemical analyzer (IDEXX, Catalyst One, USA).

[0047] Experimental results

[0048] like Figure 2 As shown, compared with the control group mice, after administration of FGF10, the level of copeptin in the mouse serum was significantly decreased (P<0.005), and the blood sodium was significantly increased (P<0.05).

[0049] Figure 2 Middle: A is the serum copeptin level of mice in each group; B is the serum sodium level of mice in each group. **P<0.005, *P<0.05, n=10.

[0050] Example 3

[0051] Safety Assessment

[0052] Experimental process

[0053] (1) Experimental animals: The same procedures as in Example 2 were performed on the mice, and the mice were kept for 1 month.

[0054] (2) Body weight detection: Measure the body weight of mice.

[0055] Experimental results

[0056] like Figure 3 As shown, the mice did not experience weight loss after administration (P>0.05).

[0057] Figure 3 Chinese: n = 12.

[0058] Example 4

[0059] Experimental process (AVP is mainly produced by the central nervous system and distributed mainly in the paraventricular hypothalamic nucleus (PVH) and the supraoptic nucleus (SON) of the hypothalamus. It was previously found that administering FGF10 into the nasal olfactory cavity can reduce AVP, so it is speculated that FGF10 acts on AVP-positive neurons, resulting in a decrease in the release of AVP by AVP-positive neurons. Therefore, electrophysiological experiments were used to verify the effect of FGF10 on AVP-positive neurons).

[0060] (1) Animal source: The experimental animals were 10 clean-grade male C57BL / 6J mice (7 weeks old, 20±4 g / mouse, purchased from Vital River Company).

[0061] Breeding environment requirements: good ventilation and air filtration system, quiet environment, room temperature maintained at 20℃, humidity controlled at 4-5%, ensure the litter is clean, and free access to water and food.

[0062] (2) Experimental grouping: The firing frequency of magnocellular neurons (AVP-positive neurons) in the PVH brain region was recorded, and then the firing frequency of neurons after administration of FGF10 was recorded, and the results were obtained through self-control.

[0063] (3) Experimental procedures: a. Brain slice preparation: 8-10 week old WT mice were perfused with cold oxygenated high-glucose slicing solution (234 mM sucrose, 11 mM glucose, 26 mM NaHCO3, 2.5 mM KCl, 1.25 mM NaH2PO4, 10 mM MgSO4, and 0.5 mM CaCl2) under isoflurane anesthesia and aerated with 95% O2 and 5% CO2. The mice were sacrificed by cervical dislocation and the brains were quickly removed and placed in the same slicing solution (0-4°C) for 2-3 minutes. Slices containing the PVH brain region were cut using a microtome (DTK-1000; dosaka, Kyoto, Japan) and incubated in artificial cerebrospinal fluid (aCSF) solution (37°C, osmolarity of ∼310 mOsm / L, aerated with 95% O2 and 5% CO2) for 60 minutes. b. Electrophysiological recordings: Brain slices were transferred to a recording chamber and observed using an upright microscope (Zeiss, Germany) and a 40x water-immersion objective (Olympus, Japan). During recordings, slices were continuously oxygenated with aCSF at a rate of 2-3 mL / min. Glass microelectrodes were pulled from borosilicate glass (Sutter Instruments) using a microelectrode puller (model P-87, Sutter Instrument Co.) with a resistance of 5-6 MΩ. Upon approaching the cell, positive pressure was applied to the glass microelectrode. The microelectrode clamped the cell, forming a high-resistance seal (10 MΩ to 5 GΩ). Cell membrane recordings were used to investigate the effects of FGF10 on the activity of AVP-positive neurons. During recordings of spontaneous neuronal firing, the control aCSF solution was replaced with aCSF solution containing 5 mg / mL FGF10, and observation was continued for approximately 5 minutes. Cellular recordings and analysis were performed using an Axopatch 200B amplifier (Molecular Devices, USA) and Igor software.

[0064] Experimental results

[0065] like Figure 4 As shown in Figure 3, the firing frequency of AVP-positive cells decreased significantly after administration of FGF10 (P<0.001).

[0066] Figure 4In the figure, A is a representative graph of the discharge frequency of AVP-positive cells recorded by patch clamp, and B is the discharge frequency level of AVP-positive neurons at baseline and after administration of FGF10.

[0067] Figure 4 Middle, n = 13, 13 neurons from 10 mice.

Claims

1. An application of FGF10 in the preparation of a drug for treating hyponatremia.

2. The use according to claim 1, characterized in that: The hyponatremia is caused by excessive release of arginine vasopressin in the circulation system.

3. The use according to claim 1, characterized in that: The FGF10 is used to reduce the concentration of serum copeptin, thereby increasing the blood sodium concentration in the serum.

4. The use according to claim 1, characterized in that: The FGF10 is used to regulate the abnormal increase of arginine vasopressin.

5. The use according to claim 1, characterized in that: The drug for treating hyponatremia is a drug preparation containing FGF10 or a drug preparation containing an FGF10 promoter.

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