CN112168841A

CN112168841A - Method for increasing bioavailability of nitric oxide in human

Info

Publication number: CN112168841A
Application number: CN201910607631.2A
Authority: CN
Inventors: 卢序
Original assignee: Individual
Current assignee: Individual
Priority date: 2019-07-05
Filing date: 2019-07-05
Publication date: 2021-01-05

Abstract

The present invention provides the use of chlorite or polymer-supported chlorite for the manufacture of a medicament for increasing the bioavailability of nitric oxide.

Description

Method for increasing bioavailability of nitric oxide in human

Technical Field

The invention belongs to the field of medicines, and particularly relates to a medical application of chlorite or polymer-supported chlorite.

Background

Nitric oxide was only considered an air pollutant before 1987. Until three scientists, Furchgott, Ignarro and Murad, identified nitric oxide as an endothelium-derived diastolic factor, they did not recognize nitric oxide anew. A great deal of research shows that nitric oxide plays an important role in the blood vessel metabolism, nerve and immune processes of human bodies.

Nitric oxide is produced in humans primarily by the breakdown of arginine by nitric oxide synthase. Nitric oxide bioavailability refers to the production and utilization of nitric oxide endogenous to the human body. A reduction in nitric oxide bioavailability can lead to a range of diseases, such as cardiovascular diseases. Strategies by increasing endogenous nitric oxide have been applied to cardiovascular, antibacterial, antitumor, wound healing, and the like treatments.

Currently, nitric oxide based therapies can be divided into two categories: one is the ability to produce nitric oxide by a substance-altering bodily enzymes; another class is redox analogues that actively release nitric oxide or nitric oxide through a substance. The first category is inflammation and toxicity caused by avoiding excessive nitric oxide production, mainly by inhibiting the activities of Inducible Nitric Oxide Synthase (iNOS) and neurogenic nitric oxide synthase (nNOS). A substance that actively increases the activity of endothelial nitric oxide synthase (eNOS) without increasing or inhibiting the activity of iNOS and nNOS would have the desirable function of increasing the bioavailability of nitric oxide. The second category was first of all small molecules such as nitroglycerin, which dilate blood vessels by metabolizing nitric oxide. However, the rate of small molecule decomposition is too fast and there is no way to release nitric oxide for a long time. At present, nucleophilic NO donors N-diazeniumdiolate and S-nitrosothiols are predominant in polymer doped nitric oxide donor systems. In addition to producing nitric oxide donors, nitrate and nitrite are also reduced to nitric oxide in the human metabolism, and are a method for improving the bioavailability of nitric oxide in the human body. Since the donor itself may have some toxicity, it is a better approach to increase the bioavailability of nitric oxide by using endogenous biochemical reactions.

Improving the content and activity of endothelial nitric oxide synthase in human body is an important path for improving the bioavailability of nitric oxide by utilizing endogenous biochemical reaction. Searles lists factors that alter the transcriptional and post-transcriptional mRNA half-life of endothelial nitric oxide synthase, indicating that hydrogen peroxide and linoleic acid oxide can increase both the transcriptional and post-transcriptional mRNA half-life of nitric oxide synthase (Searles 2006). Endothelial nitric oxide synthase, in the presence of hydrogen peroxide, also changes its phosphorylation state, thereby increasing the activity of the enzyme over a period of time, and thus increasing the amount of nitric oxide produced (Thomas, Chen,

however, not all oxidizing agents have a function of increasing the amount of nitric oxide produced at any concentration, and oxidizing agents of different oxidizing abilities have greatly different effects on the amount of nitric oxide produced. It was found that too high hydrogen peroxide can increase the amount of nitric oxide production by eNOS in the short term, but can inhibit the activity of eNOS in the long term (Hu et al.2008). Hypochlorous acid reduces eNOS nitric oxide production (Stocker et al 2004). Thus, the oxidizing properties, concentration and treatment time of the oxidizing agent can significantly affect eNOS.

Consider human blood having a pH of about 7.4 and containing a certain amount of carbon dioxide. Since hypochlorite is almost absent in an aqueous solution containing carbon dioxide, only chlorous acid is considered. The standard electrode potentials for the partial reactions in which reactions are likely to occur in blood are thus listed in table 1. Since hypochlorous acid has a high potential and inhibits eNOS, the oxidant potential must be less than 1.482V. While the potential of hydrogen peroxide is 0.878V under alkaline conditions and the potential of oxygen is 0.401V under alkaline conditions. Since oxygen does not necessarily increase the amount of nitric oxide produced by eNOS as does hydrogen peroxide. Therefore, theoretically, an oxidizing agent with a standard potential in a certain interval between 0.401 and 0.878V should be capable of generating significantly more nitric oxide from eNOS as hydrogen peroxide. Although oxidants in the interval between 0.878V and 1.482V should also perform this function, it is contemplated that strong oxidants, and weaker oxidants, are more likely to damage cellular tissue. Therefore, the lower potential section is preferred.

TABLE 1 Standard electrode potentials

In the interval of 0.401-0.878V, chlorite and chlorate are two possible oxidants that may act to stimulate eNOS. Ali et al have made a review of the effects of sodium chlorite and sodium chlorate on human red blood cells (Ali, Ahmad, and Mahmood 2017; Ali and Mahmood 2017). The experimental data show that the function of chlorite for stimulating the generation of NO by red blood cells at the same concentration is far better than that of sodium chlorate. Since the red blood cells are anucleate and contain eNOS, the effect of sodium chlorite on increasing the generation of nitric oxide by eNOS can be indirectly shown. However, Ali et al simply believe that an increase in NO represents an increase in the overall reactive oxygen species level, indicating increased damage to the tissue.

Since the mainstream view now holds that the reduction of the bioavailability of nitric oxide is often caused by the inhibition or damage of eNOS by excessive active oxygen and synergistic factors, the mainstream solution is to take in a nitric oxide sustained release donor together with the intake of a reducing agent or an active oxygen related enzyme inhibitor. Most researchers in this field believe that the ingestion of oxidants only increases the production of reactive oxygen species, exacerbating the damage caused by excess reactive oxygen species. However, since the different active oxygen species are chemically different, the function cannot be generalized and the dosage is important. In fact, it is entirely possible to have a suitable dosage range which will improve the bioavailability in humans over a long period of time without damaging the cellular tissues.

Chlorite is a substance which is safe to human body. Chlorite is less oxidizing under acidic conditions than hydrogen peroxide. Since chlorite is extremely unstable under acidic conditions or chlorous acid, it is easily decomposed into water and chlorine dioxide. In addition, chlorite does not form hydroxyl radicals as hydrogen peroxide due to the Fenton reaction (Elstner 1991). Therefore, the oxidative stress injury that chlorite causes to humans at the same concentration should be less than the active oxygen species such as hydrogen peroxide (Youngman et al 1985). One of the common food additive ingredients for chlorite is at a level of no significant damaging effect at 3mg/kg per day (Benford et al 2009), which is already well above the physiologically effective concentration.

Patent publication nos. CN102423318B, CN106413721A disclose the use of chlorite for anti-inflammatory treatment of neurodegenerative diseases. Patent publication No. CN108135930A discloses the use of chlorite for the treatment of red blood cell diseases and indications mediated thereby. However, the above patents all use periodic large doses of chlorite in excess of 1000 μmol per single intravenous injection and do not consider that low doses of oral chlorite have a significant effect on nitric oxide bioavailability in humans.

Reference to the literature

Ali，Shaikh Nisar，Mir Kaisar Ahmad，and Riaz Mahmood.2017.“Sodium Chlorate，a Herbicide and Major Water Disinfectant Byproduct，Generates Reactive Oxygen Species and Induces Oxidative Damage in Human Erythrocytes.”Environmental Science and Pollution Research 24(2)：1898-1909.https：//doi.org/10.1007/s11356-016-7980-7.

Ali，Shaikh Nisar，and Riaz Mahmood.2017.“Sodium Chlorite Increases Production of Reactive Oxygen Species That Impair the Antioxidant System and Cause Morphological Changes in Human Erythrocytes.”Environmental Toxicology 32(4)：1343-53.https：//doi.org/10.1002/tox.22328.

Benford，D.J.，F.Hill，P.Jackson，J.C.Larsen，and J.-C.Leblanc.2009.“ACIDIFIED SODIUM CHLORITE.”In Safety Evaluation of Certain Food Additives and Contaminants，edited by Food and Agriculture Organization of the United Nations and World Health Organization，3-54.

Elstner，E F.1991.“Oxygen Radicals-Biochemical Basis for Their Efficacy.”Klin.Wochenschr.69：949-56.

Hu，Zhuangli，Juan Chen，Qin Wei，and Yong Xia.2008.“Bidirectional Actions of Hydrogen Peroxide on Endothelial Nitric-Oxide Synthase Phosphorylation and Function.”Journal of Biological Chemistry 283(37)：25256-63.

https：//doi.org/10.1074/jbc.m802455200.

Searles，C D.2006.“Transcriptional and Posttranscriptional Regulation of Endothelial Nitric Oxide Synthase Expression.”Am J Physiol Cell Physiol 291(5)：C803.

Stocker，Roland，Annong Huang，Erin Jeranian，Jing Yun Hou，Tina T.Wu，Shane R.Thomas，and John F.Keaney.2004.“Hypochlorous Acid Impairs Endothelium-Derived Nitric Oxide Bioactivity through a Superoxide-Dependent Mechanism.”Arteriosclerosis，Thrombosis，and Vascular Biology 24(11)：2028-33.

https：//doi.org/10.1161/01.ATV.0000143388.20994.fa.

Thomas，Shane R.，Kai Chen，and John F.Keaney.2002.“Hydrogen Peroxide Activates Endothelial Nitric-Oxide Synthase through Coordinated Phosphorylation and Dephosphorylation via a Phosphoinositide 3-Kinase-Dependent Signaling Pathway.”Journal of Biological Chemistry 277(8)：6017-24.https：//doi.org/10.1074/jbc.M109107200.

Petr.2015.“Electrochemical Series.”In CRC Handbook of Chemistry and Physics，edited by Williams M L.，96th ed.，5-80-5-89.CRC PRESS.

Youngman，R J，G.R.Wagner，E F Elstner，and F W Kühne.1985.“Biochemical Oxygen Activation as the Basis for the Physiological Action of Tetrachlorodecaoxide(TCDO).”Zeitschrift Fur Naturforschung-Section C Journal of Biosciences 40(5-6)：409-14.

https：//doi.org/10.1515/znc-1985-5-621.

Disclosure of Invention

The present inventors have found that chlorite or polymer-loaded chlorite can significantly increase the nitric oxide production rate of endothelial nitric oxide synthase in endothelial cells, red blood cells and epithelial cells.

Thus, according to the present invention there is provided the use of chlorite or a polymer-supported chlorite for the manufacture of a medicament for increasing the bioavailability of nitric oxide in a human.

Chlorite is sodium chlorite. Polymer supported chlorite is an anion exchange resin supported chlorite. The sodium chlorite is present in the drug or drug combination in the form of sodium chlorite crystals, aqueous sodium chlorite solution, acidified sodium chlorite preparation, stable chlorine dioxide solution. The mole ratio of chlorite to chlorate is greater than 100: 5, preferably greater than 100: 1, and most preferably greater than 200: 1. The reason for setting this ratio is that chlorate in concentrations of a few micromoles per litre is sufficient to affect the sulphur metabolism in the human body, which is not required in the process of increasing the bioavailability of nitric oxide.

The user population can be: characterized by a decreased rate of nitric oxide production by endothelial nitric oxide synthase in endothelial cells or red blood cells or epithelial cells of a body part; or may be characterized by a need for an increase in the rate of nitric oxide production by endothelial nitric oxide synthase in endothelial cells or red blood cells or epithelial cells of a body part.

Chlorite or polymer-loaded chlorite can be administered alone or in combination, or in any convenient pharmaceutical form. Can be in the form of aqueous solution, or solid, and can be administered orally, intravenously, by atomization, or by external application. However, oral administration is preferred. In this application, a suitable dosage of chlorite or polymer-supported chlorite part is 1-45. mu. mol/kg per day, preferably 3-23. mu. mol/kg per day. The dose may be administered in 1 and more divided doses or in a single controlled release formulation.

Based on the present invention, chlorite or polymer-loaded chlorite can be used to increase nitric oxide bioavailability in humans. Based on example 2, ex vivo liver cell experiments (fig. 3-6), at concentrations below 50 μmol/L, no significant damage to stem cells was observed for short periods of time; at a concentration of 25. mu. mol/L, there was almost no difference from the control group at 12 hours. Based on example 3, it is shown that erythrocytes with low sodium chlorite concentration can also significantly increase the production of nitric oxide. Since the amount of NO produced by iNOS is often one to two orders of magnitude higher than that by eNOS, an increase in NO in the context of example 3 can be attributed to increased eNOS activity.

Since the only components of red blood cells that produce nitric oxide are endothelial nitric oxide synthase, chlorite is believed to increase the rate at which nitric oxide is produced by nitric oxide synthase. The liver cells are not damaged greatly below 50 mu mol/L, and endothelial cells and epithelial cells all have endothelial nitric oxide synthase. It is easily speculated that chlorite increases the rate of nitric oxide production by endothelial nitric oxide synthase in endothelial cells, epithelial cells.

Drawings

Figure 1 is a blood sample taken prior to administration of sodium chlorite.

Figure 2 is a blood sample after administration of sodium chlorite.

FIG. 3 is a photograph of a liver cell culture in an environment of 0. mu. mol/L sodium chlorite.

FIG. 4 is a photograph of a liver cell culture in an environment of 25. mu. mol/L sodium chlorite.

FIG. 5 is a photograph of a liver cell culture in an environment of 50. mu. mol/L sodium chlorite.

FIG. 6 is a photograph of a liver cell culture in an environment of 100. mu. mol/L sodium chlorite.

Detailed Description

Example 1

A45 year old woman with a perennial blood oxygen saturation of less than 95%. Blood samples were taken directly and the morphology of red blood cells is shown in FIG. 1. After 20 minutes, a blood sample was taken with 200ml of a solution containing 250. mu. mol of 95% pure sodium chlorite, and the morphology of red blood cells was shown in FIG. 2. Furthermore, the blood oxygen saturation was measured for 10, 20, 40, 60, 120 minutes after the administration of sodium chlorite for 5 consecutive days. The blood oxygen saturation at 60 and 120 minutes is significantly higher than before the day's administration.

Example 2

The isolated liver cells were cultured for a certain period of time, and then observed after being cultured for a certain period of time in the presence of 0, 25, 50, and 100. mu. mol/L sodium chlorite, and the results are shown in FIGS. 3 to 6. It was found that significant cell damage was also observed at 100. mu. mol/L, but 50. mu. mol/L damage was much reduced, with 25. mu. mol/L being almost indistinguishable from the control.

Example 3

Based on the method for treating and detecting red blood cells by Ali et al (Ali and Mahmood 2017), the influence of sodium chlorite with the concentration of less than 100 mu mol/L on the generation of nitric oxide by red blood cells of a human body is examined. Compared with a control group, the average increase amount of the nitric oxide is respectively 50% for 10 mu mol/L, 79% for 25 mu mol/L and 115% for 50 mu mol/L.

Example 4

An 88 year old male was hospitalized after a post-traumatic brain injury, and the blood oxygen saturation was measured to be 88%. The gauze was soaked in 5ml of a sodium chlorite solution with a concentration of 100. mu. mol/L and applied to the injured area of the brain. After 5 minutes the blood oxygen saturation increased to 98%. The blood oxygen saturation is stabilized at more than 96% for 2 hours. After 4 hours the blood oxygen saturation had dropped to 91% again.

The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.

Claims

1. Use of chlorite or a polymer-supported chlorite for the manufacture of a medicament for increasing the bioavailability of nitric oxide in a human.

2. Use according to claim 1, wherein the chlorite is sodium chlorite.

3. The use as claimed in claim 1, wherein the polymer-supported chlorite is anion exchange resin-supported chlorite.

4. Use according to claim 3, wherein the sodium chlorite is present in the medicament or pharmaceutical combination in the form of sodium chlorite crystals, aqueous sodium chlorite solution, acidified sodium chlorite preparation, stable chlorine dioxide solution.

5. Use according to claim 4, wherein the mole ratio of chlorite to chlorate is greater than 100: 5, preferably greater than 100: 1, most preferably greater than 200: 1.

6. The use of any one of claims 1-5, wherein the human is characterized by a decreased rate of nitric oxide production by endothelial nitric oxide synthase in endothelial cells or red blood cells or epithelial cells of the body part.

7. The use according to any one of claims 1 to 5, wherein the human is characterized by a need for an increased rate of nitric oxide production by endothelial nitric oxide synthase in endothelial cells or red blood cells or epithelial cells of the body part.

8. Use according to any one of claims 1 to 5, whereby the chlorite or polymer-loaded chlorite can be administered orally, intravenously, by nebulization, topically, preferably orally.

9. Use according to claim 8, wherein the chlorite or polymer-loaded chlorite part is suitably administered in a dose of 1-45 μmol/kg per day, preferably 3-23 μmol/kg per day.

10. The use of claim 9 wherein the chlorite or polymer-loaded chlorite root partial doses are administered in 1 and more partial doses or in a single controlled release formulation.