Application of long-chain fatty acid 12-HHT in sleep apnea complicated with hypertension

Abstract

This invention discloses the application of the long-chain fatty acid 12-HHT in sleep apnea complicated with hypertension, belonging to the field of biomedicine. Specifically, 12-HHT is used as a diagnostic or drug target to prepare drugs or reagents for the diagnosis or treatment of sleep apnea complicated with hypertension. The molecular formula of 12-HHT is C2. 17 H 28 O3, with the structural formula shown, refers to obstructive sleep apnea-hypopnea syndrome combined with hypertension. The drug can be a targeted therapy, with 12-HHT as the target. This invention, using an intermittent hypoxic endothelial cell model, found that 12-HHT can inhibit excessive proliferation and migration of endothelial cells induced by intermittent hypoxia, thus exerting a therapeutic effect. This demonstrates its effectiveness as an intervention target for OSAHS combined with hypertension, providing a novel potential therapeutic drug for this type of patient.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to the application of long-chain fatty acid 12-HHT in sleep apnea complicated with hypertension. Background Technology

[0002] Obstructive sleep apnea-hypopnea syndrome (OSAHS) is a systemic disease in which the upper airway is partially or completely blocked during sleep at night, causing repeated episodes of apnea and hypopnea, which can eventually lead to cardiovascular and cerebrovascular complications.

[0003] OSAHS (Obesory-Spoken Hypoxia) is one of the most common causes of secondary hypertension, and the severity of OSAHS is often positively correlated with the degree of blood pressure elevation. Currently, for patients with both sleep apnea and hypertension, medication alone is less effective in lowering blood pressure, and non-invasive positive airway pressure (NPAP) has a limited impact on blood pressure levels in these patients. Even with the addition of one or more antihypertensive drugs to this treatment, cardiovascular events and death cannot be completely prevented. Therefore, addressing the poor treatment outcomes of sleep apnea combined with hypertension requires early diagnosis and optimization of existing drug treatment methods. Summary of the Invention

[0004] To overcome the problem of poor antihypertensive efficacy in the treatment of OSAHS combined with hypertension in existing technologies, the purpose of this invention is to provide the application of long-chain fatty acid 12-HHT as a diagnostic or drug target in the preparation of drugs or reagents for the diagnosis or treatment of sleep apnea combined with hypertension. Long-chain fatty acid 12-HHT is a metabolite of arachidonic acid via the cyclooxygenase (COX) pathway, a low-affinity ligand of BLT2, with CAS number 100679-07-0 and molecular formula C2. 17 H 28 O3, with a molecular weight of 280.40, has the following structural formula:

[0005] Preferably, the sleep apnea combined with hypertension refers to obstructive sleep apnea-hypopnea syndrome combined with hypertension.

[0006] The present invention also provides a pharmaceutical composition, which is a targeted drug with long-chain fatty acid 12-HHT as the target.

[0007] This invention also provides the use of long-chain fatty acid 12-HHT or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating sleep apnea complicated with hypertension, wherein the molecular formula of long-chain fatty acid 12-HHT is C 17 H 28 O3, structural formula is

[0008] Preferably, the sleep apnea combined with hypertension refers to obstructive sleep apnea-hypopnea syndrome combined with hypertension.

[0009] Preferably, the pharmaceutically acceptable salt of the long-chain fatty acid 12-HHT is selected from one or more combinations of hydrochloride, hydrobromide, sulfate, acetate, lactate, tartrate, tannate, citrate, trifluoroacetate, malate, maleate, succinate, p-toluenesulfonic acid, or benzenesulfonate.

[0010] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of long-chain fatty acid 12-HHT and / or a pharmaceutically acceptable salt thereof, wherein the molecular formula of long-chain fatty acid 12-HHT is C 17 H 28 O3, structural formula is

[0011] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient, and can be prepared as an injection, emulsion, tablet, powder, granule, ointment, liposome, or oral liquid.

[0012] This invention also provides a small molecule injectable formulation containing a therapeutically effective amount of the long-chain fatty acid 12-HHT with the molecular formula C 17 H 28 O3, structural formula is The concentration of the long-chain fatty acid 12-HHT is 0.1-100 μmol / L, preferably 3 μmol / L.

[0013] Previous literature has reported that 12-HHT has functions such as regulating allergic reactions and repairing the epithelial barrier. This invention demonstrates, through serum metabolomics analysis in humans and rats, that 12-HHT is correlated with obstructive sleep apnea-hypoxia (OSAHS) combined with hypertension. Furthermore, experiments using an intermittent hypoxic endothelial cell model revealed that 12-HHT can exert a therapeutic effect on this cell model through anti-excessive migration, suggesting it could be used as an effective intervention target for OSAHS combined with hypertension in the preparation of drugs for treating sleep apnea-hypoxia with hypertension, optimizing existing drug treatments and providing novel potential therapeutic agents for these patients. In addition, 12-HHT is an endogenous fatty acid with good drug safety. Attached Figure Description

[0014] Figure 1 The relative abundance of 12-HHT in the serum of patients, healthy individuals, and rats in each group of examples is shown.

[0015] Figure 2The following is an example of the effect of 12-HHT on the proliferation phenotype of intermittent hypoxic endothelial cells: A: EdU detection results of human primary umbilical vein endothelial cells (HUVECs) in each group; B: Statistical graph of EdU-positive cells in HUVECs in each group.

[0016] Figure 3 The following is an example of the effect of 12-HHT on the migration phenotype of intermittent hypoxic endothelial cells (scratch assay); A: Scratch assay results of HUVECs in each group; B: Statistical graph of healing rate of HUVECs in each group.

[0017] Figure 4 The following is an example of the effect of 12-HHT on the migration phenotype of intermittent hypoxic endothelial cells (transwell assay); A: Transwell staining results of HUVECs in each group; B: Statistical graph of the number of migrating HUVECs in each group. Detailed Implementation

[0018] The present invention will be further described in detail below with reference to embodiments, but the present invention is not limited to these embodiments.

[0019] Example 1

[0020] Newly diagnosed hypertensive patients (HTN group), OSAHS patients (OSAHS group), and OSAHS combined with hypertension (OSAHS+HTN group) who visited the cardiology and respiratory medicine outpatient clinics of Zhongshan Hospital affiliated with Fudan University between January 2021 and October 2022 were selected. Additionally, a clinically matched population who visited the health check-up center during the same period were included as a healthy control group (CON group). The control group was excluded if hypertension and OSAHS were present. Serum non-targeted metabolomics were performed on all subjects. Simultaneously, rat models of intermittent hypoxia (Osahs group), intermittent hypoxia combined with hypertension (Osahs+Htn group), and a control group (Con group) were established, benchmarked against clinical samples (to distinguish them from clinical samples, rat grouping in the first part of the study used "uppercase + lowercase" format). Serum non-targeted metabolomics of the rats was also measured and compared with the results of clinical patients. Specific procedures and results are as follows:

[0021] (1) Relative abundance of serum 12-HHT in each group of patients and healthy individuals: A total of 125 patients who visited the Department of Cardiology and Department of Respiratory Medicine at Zhongshan Hospital affiliated with Fudan University from January 2021 to October 2022 were included, including 40 patients in the HTN group, 30 patients in the OSAHS group, 55 patients in the OSAHS+HTN group, and 38 patients in the CON group from the health check-up center. Serum metabolomics tests were performed on the above populations, and statistically significant differences were found in the pairwise comparisons of the OSAHS group, the OSAHS+HTN group, and the CON group.

[0022] (2) Relative abundance of serum 12-HHT in each group of rats: Male Wistar rats were divided into three groups: Osahs experimental group (n=5, 8 weeks old), Osahs+Htn experimental group (n=4, 8 weeks old), and Con group (n=5, 8 weeks old). During the daytime (9 am - 7 pm, 10 h) when the rats' sleep phase was dominant, the experimental groups were subjected to continuous intermittent hypoxia stimulation. The oxygen partial pressure was set within a range of approximately 6.5%-21%, with one cycle lasting 220 seconds. Starting with the hypoxic period, oxygen was introduced into the animal chamber for 25 seconds, causing the oxygen concentration to slowly rise to 21%, and maintained at 21% for 90 seconds. Then, nitrogen was introduced into the animal chamber for 45 seconds, causing the oxygen concentration to slowly decrease to 6.5%, and maintained at 6.5% for 60 seconds. This cycle was repeated. During the plateau phase of normoxic or hypoxic conditions, the programmable controller automatically replenished the oxygen consumed by the rats' vital activities to ensure that the oxygen concentration was maintained stably at approximately 21% or 6.5%. The intermittent hypoxic stimulation time was 4 hours on day 1 of modeling, increased to 7 hours on day 2, and 10 hours from day 3 onwards until the end of modeling. If carbon dioxide levels exceeded 1% in the hyperoxic and hypoxic chambers during the experiment, soda lime was added for absorption. The Osahs experimental group received intermittent hypoxic stimulation for 2 weeks, and the Osahs+Htn group received intermittent hypoxic stimulation for 8 weeks. The Con group maintained a normoxic partial pressure (21%) throughout the experiment, with other conditions unchanged for a total of 8 weeks. Serum metabolomics analysis of the rats in each group revealed statistically significant differences in 12-HHT levels between all groups.

[0023] Metabolomics results in humans and rats are as follows Figure 1 As shown, serum levels of the long-chain fatty acid 12-HHT in both humans and rats exhibited significant and consistent changes in both the OSAHS group and the OSAHS combined with hypertension group. Furthermore, metabolomics results from the patients revealed no significant difference in 12-HHT levels between the CON and HTN groups. This demonstrates that 12-HHT is significantly associated with OSAHS combined with hypertension and could serve as a potential biomarker for this condition.

[0024] Example 2

[0025] This embodiment constructs an intermittent hypoxic endothelial cell model to verify the effects of 12-HHT on the proliferation and migration phenotypes of intermittent hypoxic endothelial cells. The specific operation and results are as follows:

[0026] Human primary umbilical vein endothelial cells (HUVECs) were divided into four groups: normotrophic group (Nx group), normoxic + 12-HHT group (Nx + 12-HHT group), intermittent hypoxia group (IH group), and intermittent hypoxia + 12-HHT group (IH + 12-HHT group). Cell proliferation was analyzed using the EdU assay, and cell migration was evaluated using scratch assay and transwell assay. The intermittent hypoxia model was as follows: a 50-minute cycle was performed, with binary gas introduced into the cell culture chamber for 2 minutes, allowing the oxygen concentration to rise slowly and then decrease to approximately 1%, maintaining a 1%-2% oxygen concentration for 15 minutes. Subsequently, a ternary gas mixture was introduced into the cell culture chamber for 3 minutes, allowing the oxygen concentration to rise slowly to approximately 21%, maintaining a 20%-21% oxygen concentration for 30 minutes. This cycle was repeated for 24 hours to establish the intermittent hypoxia model. The normoxic control group maintained a gas environment of 21% O2, 5% CO2, and the remainder N2 during the experiment, with all other conditions identical to the intermittent hypoxia group.

[0027] The results are as follows Figures 2 to 4 As shown, the addition of 3 μmol / L 12-HHT to an intermittent hypoxic endothelial cell model revealed that 12-HHT could inhibit the excessive proliferation and migration of HUVECs induced by intermittent hypoxia. This demonstrates that 12-HHT can serve as an effective intervention target for OSAHS combined with hypertension, providing a novel potential therapeutic drug for such patients and possessing significant clinical application value.

[0028] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The use of long-chain fatty acid 12-HHT in the preparation of drugs for treating sleep apnea complicated with hypertension, wherein the molecular formula of long-chain fatty acid 12-HHT is C 17 H 28 O3, structural formula is The sleep apnea combined with hypertension refers to obstructive sleep apnea-hypopnea syndrome combined with hypertension.

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