A method for preparing an anti-aging or longevity-extending drug

Abstract

Omeprazole, as a widely used proton pump inhibitor, is mainly used for the treatment of various diseases caused by excessive gastric acid, such as gastric ulcer, duodenal ulcer and reflux esophagitis. The present application relates to a new use of a commercially available drug omeprazole, i.e. it has significant anti-aging activity. The present application found that omeprazole can significantly prolong the life of nematodes through research using model organisms Caenorhabditis elegans. Specifically, when the nematodes are cultured in a medium containing 1.6 mg / mL and 3.2 mg / mL omeprazole, their life is prolonged by 47.80% and 61.51% respectively, which is extremely rare in current drug research.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to a novel method for preparing anti-aging or life-extending drugs using proton pump inhibitors. Background Technology

[0002] Omeprazole (chemical name: 5-methoxy-2-[[(4-methoxy-3,5-dimethyl-2-pyridyl)methyl]sulfinyl]-1H-benzimidazole, generic name: Omeprazole) is a proton pump inhibitor, a lipid-soluble weakly basic drug. It readily concentrates in acidic environments and specifically acts on the secretory microtubules formed by the apical membrane of gastric parietal cells and the tubular vesicles in the cytoplasm, the location of the gastric parietal cell proton pump (H+,K+-ATPase). It is converted to its active form, sulfinamide, and irreversibly binds to the sulfhydryl group of the proton pump via disulfide bonds, thereby inhibiting the activity of H+,K+-ATPase, blocking the final step of gastric acid secretion, preventing H+ from being transported from the parietal cells into the gastric lumen, and greatly reducing the acid content in gastric juice. It has a strong inhibitory effect on both basal gastric acid and irritated gastric acid secretion. Omeprazole significantly inhibits gastric acid secretion induced by histamine, pentagastrin, and vagal nerve stimulation. It also exhibits a strong and sustained inhibitory effect on gastric acid secretion induced by butylated cyclic adenosine monophosphate (bcAMP), which cannot be inhibited by H2 receptor antagonists. Following administration, gastric pH rapidly increases with a significant decrease in gastric acid secretion; it also provides rapid relief from heartburn and pain. Furthermore, it has a high cure rate for duodenal ulcers and a low recurrence rate.

[0003] Acute toxicity studies showed that the LD50 of omeprazole in rats was 2210 mg / kg orally and >50 mg / kg intravenously; in mice, it was >4 mg / kg orally and 82.8 mg / kg intravenously. Developmental or reproductive toxicity studies showed that omeprazole at 345 times the human dose had no teratogenic effect in pregnant rats. Genotoxicity studies showed no evidence of mutagenicity with omeprazole; bacterial reverse mutation assays, in vitro mouse lymphoma cell assays, in vivo assays, and rat liver DNA damage tests were negative. Mouse micronucleus assays and chromosomal aberration assays yielded indeterminate results; however, other reports indicate a negative mouse micronucleus assay. These experimental data demonstrate that omeprazole has good safety and extremely low acute toxicity and genotoxicity.

[0004] Developing new indications for known drugs is a crucial aspect of drug development. Given omeprazole's good safety profile and extremely low acute and genotoxic effects, it provides a solid foundation for researching and developing other indications. European patent EP0197013A1 discloses a method for treating inflammatory bowel diseases using omeprazole and its analogues; PCT patent WO8903829A1 discloses a method for treating osteoporosis, primary and secondary hyperparathyroidism, and related diseases using omeprazole; PCT patent WO9518612A1 discloses a method for treating psoriasis using omeprazole and its analogues; PCT patent WO2006118534A1 discloses a method for treating sleep disorders caused by silent gastroesophageal reflux using proton pump inhibitors, including omeprazole; and PCT patent WO2005077365A2 discloses a method for treating or preventing cancer using proton pump inhibitors. The above-mentioned prior art disclosures demonstrate the broad application prospects of proton pump inhibitors such as omeprazole in the development of new indications.

[0005] In the fields of drug research and anti-aging, people have been searching for effective ways to slow down the aging process. Although many drugs and treatments have been studied, most suffer from limited efficacy, significant side effects, or high costs. Omeprazole, a widely used proton pump inhibitor, is primarily used to treat conditions caused by excessive stomach acid. However, its potential role in anti-aging has not yet been thoroughly investigated. Summary of the Invention

[0006] The main objective of this invention is to overcome the shortcomings of the prior art and provide a new use or indication for the commercially available drug omeprazole. This new use or indication has not been disclosed in the prior art, thus further expanding the application field of omeprazole.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] The present invention provides the use of a proton pump inhibitor in the preparation of an anti-biological aging drug, wherein the proton pump inhibitor is preferably selected from omeprazole or a pharmaceutically acceptable salt thereof.

[0009] The use of a proton pump inhibitor in the preparation of a biological life-extending drug is provided, wherein the proton pump inhibitor is preferably selected from omeprazole or a pharmaceutically acceptable salt thereof.

[0010] Furthermore, the present invention provides the use of a proton pump inhibitor in the preparation of anti-aging drugs or life-extending drugs, wherein the organism is selected from animals.

[0011] Furthermore, the present invention provides the use of a proton pump inhibitor in the preparation of an anti-aging drug or a life-extending drug, wherein the organism is selected from mammals, preferably humans.

[0012] Furthermore, the present invention provides the use of a proton pump inhibitor in the preparation of anti-aging drugs or life-extending drugs, wherein the organism is selected from nematodes, preferably from Caenorhabditis elegans (C. elegans).

[0013] Furthermore, the present invention provides the use of a proton pump inhibitor in the preparation of anti-aging drugs or life-extending drugs, wherein the concentration of the proton pump inhibitor is greater than 1 mg / mL, preferably greater than 1.2 mg / mL, and more preferably greater than 1.6 mg / mL.

[0014] Furthermore, the present invention provides the use of a proton pump inhibitor in the preparation of anti-aging drugs or life-extending drugs, wherein the concentration of the proton pump inhibitor is 1.6–3.2 mg / ml.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] This invention, through research using the model organism *C. elegans*, has found that omeprazole can significantly prolong the lifespan of nematodes, an effect that is extremely rare in current drug research.

[0017] This invention opens up new application areas for known drugs and lays a good foundation for the development of anti-aging drugs. Attached Figure Description

[0018] Figure 1 This study shows the results of a study on the lifespan extension of wild-type nematode N2 by using 0.8 mg / mL omeprazole.

[0019] Figure 2 This study shows the results of a study on the lifespan extension of wild-type nematode N2 by 1.6 mg / mL omeprazole.

[0020] Figure 3 This study shows the results of a study on the lifespan extension of wild-type nematode N2 by 3.2 mg / mL omeprazole.

[0021] Figure 4 The results of a comprehensive study showed that omeprazole extended the lifespan of wild-type nematodes, with the positive control group being 50 mM metformin [1].

[0022] Figure 5 Omeprazole reduces lipofuscin levels in wild-type nematodes.

[0023] Figure 6 Omeprazole prolongs the lifespan of wild-type nematodes without relying on the insulin signaling pathway.

[0024] Figure 7 Omeprazole prolongs the lifespan of wild-type nematodes without relying on dietary restriction signaling pathways.

[0025] Figure 8 Omeprazole prolongs the lifespan of wild-type nematode Nematode I in mitochondrial respiratory chain complex I.

[0026] Figure 9 Omeprazole prolongs the lifespan of wild-type nematodes without dependence on mitochondrial respiratory chain complex II.

[0027] Figure 10 Omeprazole prolongs the lifespan of wild-type nematodes without dependence on mitochondrial respiratory chain complex III.

[0028] Figure 11 Omeprazole reduces ROS levels in wild-type nematodes. Detailed Implementation

[0029] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below in conjunction with specific embodiments.

[0030] Caenorhabditis elegans (C. elegans) is a non-toxic, harmless nematode that can survive independently. The signaling pathways that regulate aging in nematodes, namely the insulin / insulin-like growth factor 1 (IGF-1) signaling pathway (IIS) and the target of rapamycin (TOR) signaling pathway, are highly conserved in mammals [2-8]. The ability of insulin receptor mutations to prolong lifespan was first discovered in nematodes [9], and dietary restriction in C. elegans also prolongs the lifespan of nematodes

[10] . In addition, sirtuin signaling, JNK signaling pathway, and TUBBY signaling pathway are also involved in the regulation of nematode lifespan. The conservatism of these signaling pathways makes nematodes an ideal model for anti-aging research. Since 1965, scientist Sydney Brenner has used nematodes as a model organism in the fields of molecular biology and developmental biology.

[0031] This invention uses *Caenorhabditis elegans* as a model organism to study the anti-aging effects of omeprazole.

[0032] The omeprazole used in this study was purchased from Yuekang Pharmaceutical Group Co., Ltd., under the brand name Yuekang Liweike.

[0033] Example 1

[0034] (1) Drug information:

[0035] Name: Omeprazole Enteric-coated Capsules - Yuekang Liweike

[0036] Manufacturer: Yuekang Pharmaceutical Group Co., Ltd.

[0037] Product batch number: 143240732

[0038] Production date: July 29, 2024

[0039] Valid until: July 28, 2026

[0040] (2) Administration method:

[0041] 1) Drug preparation:

[0042] 8 mg, 16 mg, and 32 mg of the drug were taken respectively, and 1 mL of dd H2O was added to each to obtain drug concentrations of 8 mg / mL, 16 mg / mL, and 32 mg / mL. During the experiment, the drug was injected at one-tenth of the total volume of the culture medium, and the final concentrations of the drug for the lifetime experiment were 0.8 mg / mL, 1.6 mg / mL, and 3.2 mg / mL, respectively.

[0043] 2) Culture medium preparation:

[0044] Inside the laminar flow hood, add the sterilized NGM medium to a 6cm petri dish;

[0045] 3) Nematode culture:

[0046] In the drug group, 432 μL of NGM mix medium and 48 μL of the drug at concentrations of 8 mg / mL, 16 mg / mL, or 32 mg / mL were added to 24-well plates, respectively. In the negative control group, the drug was replaced with 48 μL of dd H2O, and in the positive control group, 48 μL of 500 mM metformin was added. [1] Each group was run in 12 replicates. 10-12 young N2 wild-type adults were picked into each replicate, sealed with sealing film, and placed in an incubator at 20℃. The lifespan of the nematodes was recorded daily; the mortality rate of the nematodes was recorded every two days, which is the lifespan experiment of the nematodes.

[0047] 4) Observe and record the effect of omeprazole on the lifespan of nematodes:

[0048] The mortality rate of nematodes was counted every two days to obtain their lifespan. Experimental results showed that omeprazole significantly prolonged the lifespan of *C. elegans*. Nematodes cultured at concentrations of 0.8 mg / mL, 1.6 mg / mL, and 3.2 mg / mL showed that nematodes cultured at concentrations of 1.6 mg / mL and 3.2 mg / mL experienced lifespan extensions of 47.80% and 61.51%, respectively, as detailed in Table 1.

[0049] Table 1. Omeprazole prolongs the lifespan of wild-type nematode N2.

[0050]

[0051] Omeprazole reduces lipofuscin levels in wild-type nematodes.

[0052] Wild-type nematodes were cultured for 10 days in omeprazole at concentrations of 0.8 mg / mL, 1.6 mg / mL, or 3.2 mg / mL. The lipofuscin fluorescence level of the nematodes was detected at an excitation wavelength of 340 nm. The results showed that the lipofuscin levels in the nematodes cultured at all three concentrations were significantly lower than those in the control group. Figure 5 As shown.

[0053] Omeprazole prolongs the lifespan of wild-type nematode Nematode without relying on the insulin signaling pathway.

[0054] Daf-2 is a homolog of the insulin / IGF-1 receptor in nematodes, and the CB1370 strain is... daf-2 A mutant strain with a mutated gene. When this mutant was cultured with 1.6 mg / mL omeprazole, its lifespan was tested. The results showed that the CB1370 nematode strain cultured with 1.6 mg / mL omeprazole had a significantly longer average lifespan compared to the control group. Figure 6 As shown, epistatic analysis using genetic methods concludes that the anti-aging effects of omeprazole may not depend on the insulin signaling pathway.

[0055] Omeprazole prolongs the lifespan of wild-type nematode Nematode without relying on dietary restriction signaling pathways.

[0056] nematode strain DA465 is eat-2 This gene mutant, exhibiting impaired pharyngeal function and thus reduced food intake, is a recognized nematode model in dietary restriction research. We cultured DA465 nematodes in medium containing 1.6 mg / mL omeprazole and determined their lifespan. The results showed that omeprazole treatment significantly prolonged the lifespan of the DA465 strain of nematodes; detailed data can be found in [link to data]. Figure 7 This finding suggests that the anti-aging effect of omeprazole on nematodes is not dependent on the dietary restriction signaling pathway.

[0057] Omeprazole prolongs the lifespan of wild-type *C. elegans* in mitochondrial respiratory chain complex I.

[0058] MQ1333 is a mutant of the nuo-6 gene in nematodes, which impairs the function of mitochondrial respiratory chain complex I. Treatment of this mutant with 1.6 mg / mL omeprazole and subsequent lifespan testing revealed that the average survival of the MQ1333 nematode strain treated with 1.6 mg / mL omeprazole was significantly shorter than that of the control group. Figure 8 As shown in the figure. This finding suggests that the anti-aging effect of omeprazole may depend on the function of this mitochondrial respiratory chain complex I.

[0059] Omeprazole prolongs the lifespan of wild-type *C. elegans* independently of mitochondrial respiratory chain complex II.

[0060] TK22 is a mutant of the mev-1 gene, which encodes the cytochrome b subunit of succinate dehydrogenase (SDHC), a key component of mitochondrial respiratory chain complex II.

[27] MEV-1(kn1) mutations lead to impaired function of complex II, hindered electron transport, and consequently, excessive production of superoxide anions (O2⁻) in mitochondria, exacerbating oxidative stress.

[29] .

[0061] TK22 mutants were cultured with 1.6 mg / mL omeprazole, and lifespan experiments were conducted. The results showed that the TK22 nematode strain treated with 1.6 mg / mL omeprazole had a significantly longer average lifespan compared to the control group. Figure 9 As shown in the figure. This result suggests that the anti-aging effect of omeprazole may not depend on the function of mitochondrial respiratory chain complex II.

[0062] Omeprazole prolongs the lifespan of wild-type *C. elegans* independently of mitochondrial respiratory chain complex III.

[0063] MQ887 is a mutant of the isp-1 gene, which encodes the Rieske iron-sulfur protein in mitochondrial respiratory chain complex III. The mutation affects the function of mitochondrial respiratory chain complex III.

[0064] The MQ887 mutant strain was cultured with 1.6 mg / mL omeprazole, and a lifespan test was conducted. The results showed that 1.6 mg / mL omeprazole significantly prolonged the lifespan of the MQ887 mutant. Detailed results can be found in [link to results]. Figure 10 This result suggests that the anti-aging effects of omeprazole may not depend on the function of mitochondrial respiratory chain complex III.

[0065] Omeprazole reduces ROS levels in wild-type nematodes.

[0066] Wild-type nematodes were cultured with 0.8 mg / mL omeprazole for 72 hours, then treated with 10 mM potassium dichromate oxidative stress for 48 hours. ROS levels were then detected using a DCFDA fluorescent probe.

[32] It was found that 0.8 mg / mL omeprazole significantly reduced the fluorescence intensity of nematodes, suggesting that omeprazole may enhance antioxidant and anti-aging capabilities by reducing ROS levels. (See results below.) Figure 11 .

[0067] Some references to this invention:

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[0085] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. Use of a proton pump inhibitor in the preparation of a drug for prolonging the lifespan of an organism, wherein the proton pump inhibitor is omeprazole or a pharmaceutically acceptable salt thereof, and the organism is wild-type Caenorhabditis elegans; the concentration of the proton pump inhibitor is 1.6–16 mg / ml.

2. The use according to claim 1, characterized in that... The concentration of the proton pump inhibitor is 1.6–3.2 mg / ml.

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