RU2713153C1 - Method of inhibiting tumour growth in mammal - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine and can be used for tumour growth inhibition in a mammal. That is ensured by using agent containing 20 % fat lipofundin emulsion, xenon saturated with inert gas, propranolol beta blocker, reserpine sympatholytic, ivabradine antianginal preparation, an antihistamine preparation of diphenhydramine, a neuroleptic preparation with hypothermic action of Periciazine, serotoninergic agent serotonin hydrochloride, an antithyroid agent Propycil, magnesium sulphate and a pharmaceutically acceptable solvent, and a second preparation which contains 20 % lipofundin fat emulsion, xenon-saturated inert gas, wherein 1 ml of 20 % fat lipofundin emulsion contains 0.54 ml to 2.2 ml of xenon at saturation under pressure of 2 atm. Agent is injected into a mammal, after which the mammal is exposed to additional hypoxia by reducing oxygen in the external medium to level of 5 % and maintained in these conditions for 1.5–3 hours.

EFFECT: group of inventions provides intensified therapeutic effect ensured by synergetic action of said agents and subsequent use of external hypoxia on inhibition of tumor growth and regression at initial stage of growth.

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Description

Technical field

The invention relates to medicine and can be used to treat cancer, including inhibition of tumor growth, which allows to increase the time during which, if necessary, surgical treatment can be carried out or chemotherapy can be used.

State of the art

Oncological diseases are by far the most common pathology in the world after stroke and myocardial ischemia. Due to the heterogeneity of the nature and course of this disease, an active search is currently underway for new methods for the diagnosis and treatment of oncologies of various origins.

In addition to surgical methods, chemotherapy and immunotherapy, there are alternative methods of treating cancer, which include carrying out hyperthermia of affected organs, using hypothermia of organs, using gas mixtures to oxygenate diseased organs, or using methods of creating hypoxia. Such a wide range of effects is caused by the fact that until now all the features of the formation of tumors in the body of mammals and the possibility of influencing the growth of malignant tumors have not been studied.

It is known that malignant neoplasms lead to the development of tissue oxygen deficiency (hypoxia) or are directly associated with acute or chronic hypoxia. Hypoxia is a decrease in oxygen tension in body tissues (pO2) due to a violation of its delivery or disposal. In patients with malignant neoplasms, tissue hypoxia can occur for various reasons - this is a violation of the patency of the vessels supplying the organ with blood (atherosclerosis, inflammation, edema, etc.), and a decrease in the amount of hemoglobin, and many other reasons. Occurring primary or secondary, hypoxia leads to a cascade of reactions that aggravate the course of the underlying disease, which further complicates oxygen deficiency. Hypoxia ultimately leads to energy deficiency of cells - a universal outcome of almost all forms of its pathology. To treat this condition, various methods of oxygen therapy (oxygen therapy) have been developed. However, under normal atmospheric pressure, even breathing with pure oxygen is often not able to eliminate oxygen deficiency at the level of cells of organs and tissues.

Known RF patent No. RU 2481091 A METHOD FOR TREATING ONCOLOGICAL LUNG CANCER PATIENTS (05/10/2013) in which an increase in treatment effectiveness is achieved by subjecting the patient to hyperbaric oxygenation at a pressure of 1.3 atm for 30 minutes every day for 5 days before administering chemotherapy drugs. Moreover, on the first day, the patient is injected with co-plate 75 mg / m ² of the patient’s body, diluted in 0.9% sodium chloride solution, on the first, third and fifth day, the patient is also given intravenous phytoside in a dose of 120 mg / m ² of the patient’s body, diluted in 0.9% sodium chloride solution.

Known RF patent No. RU 2414936 METHOD FOR TREATING RECTAL GUT CANCER, which proposes a new method for treating colorectal cancer by exposing the tumor to remote radiation therapy, local microwave hyperthermia, the use of metronidazole as a part of a composite mixture administered intrarectally, capecitabine used orally for the entire course of radiation therapy, oxaliplatin administered intravenously, the use of a hypoxic gas mixture containing 9% oxygen and 91% nitrogen to protect surrounding tissues from exposure to radiation from the next surgical intervention. When used together, these agents have a synergistic effect and enhance the therapeutic effect due to the implementation of the mechanisms of radiosensitization and chemosensitization and synergism of radiosensitizing agents.

Known RF patent No. RU 2326701 METHOD FOR TREATING ONCOLOGICAL PATIENTS AND A DEVICE FOR ITS IMPLEMENTATION. According to the description of the method, an alternation of hypo- and hyperoxic breathing patterns of the patient is carried out using a corrugated breathing tube device. For 2-3 weeks, the latent and compensated hypoxia regimen alternates with hyperoxia, for this a controlled process of therapeutic effect on the patient’s body is created by various ratios of the gas composition of the inhaled air (oxygen from 5% to 100%, carbon dioxide from 0.03% to 5% ) with the simultaneous inhalation of drugs (sodium hypochlorite).

The use of hypothermia for the treatment of cancer is described in US patent No. US 6736837. The invention relates generally to methods for treating cancer and other diseases by modulating body temperature. Heat can be directed to the hypothalamus of a warm-blooded animal to cool the animal using physiological mechanisms that regulate body temperature to compensate for compensatory cooling, thereby lowering body temperature (hypothermia) and making other methods of lowering body temperature more effective. Heat can be removed from the animal’s hypothalamus, cooling the hypothalamus, causing a compensatory increase in body temperature (hyperthermia) and making other methods of increasing body temperature more effective. Body temperature can be directly modulated by a heat exchange catheter located inside the patient's blood vessel. The invention relates generally to methods for treating cancer by inducing hypothermia by directing heat to the hypothalamus, maintaining cancer tissue if necessary at or near normal body temperature, and optionally using another cancer treatment.

Induction of the state of stupor (torpor) is described in application for US invention No. US 20070213295. The invention relates to the detection of 5'-AMP and its analogues, which can be used to induce a state of stupor in subjects, as evidenced by studies conducted in laboratory mice. In these studies, mice were injected with high doses of 5'-AMP, which were found to unleash a mechanism for regulating the body temperature of animals, accompanied by a decrease in the temperature of the body of the animal nucleus, which, as a rule, decreased to the ambient temperature. It is proposed to use the invention for the treatment of cancer.

Known US patent No. US 7993681, which describes several methods of treating cancer by inducing numbness. Root induction may result from exposure to chalcogenide, exposure to a low oxygen environment, and / or exposure to one or more hypothermic treatments. Each of these methods of treatment can be combined with additional therapies to further form the stress of the tumor cells, including reducing blood flow to the tumor cells by extracting part of the blood to the area surrounding the tumor cell, as well as reducing the blood flow to the tumor cells by removing part of the blood stream from the specified patient and / or treatment with conventional chemotherapeutic agents.

The proposed technical solutions have several disadvantages. In particular, they involve one of the mechanisms for increasing nonspecific resistance to tumor development, realized through hypoxia or hypothermia.

Due to the rather complex mechanism of the body’s reaction to the development of the tumor and the insufficiently elaborated question about the effect of hypoxia on the state of the body in the early stages of tumor development, its use, as can be seen from the prior art, is limited to the already developed tumor.

The same can be said about the effect of hypothermia.

Disclosure of the invention

The objective of the invention is the creation of a new method of inhibiting tumor growth, the use of which will achieve regression of the tumor at the initial stage of its growth for subsequent radical surgery.

The problem is solved by the fact that a new method of inhibiting the growth of tumors at the first stage of their occurrence by a combined effect on the tumor is proposed. The combined use of a means to protect against hypoxia of the tumor carrier organism and the effect of an external hypoxic environment with a low oxygen level can provide a synergistic effect on the inhibition of tumor growth and enhance the therapeutic effect.

The method includes the formation of hypometabolism and hypothermia, in which a means is used to form hypothermia and hypometabolism, which includes a fat emulsion saturated with an inert gas, an adrenergic blocker, a sympatholytic, an histamine H1 receptor antagonist, an inhibitor of If-channels of the sinus node, a serotonergic agent, a neuroleptic agent with hypothermic effect, antithyroid drug, a preparation containing magnesium ions, which is administered to the mammal, while after administration of the agent, the mammal undergoes ayut additional hypoxia by lowering the oxygen in the external medium to a level of 5% for 35-40 minutes, and holding in these hypoxic conditions for 1.5 - 3 hours.

The tool is a combination of drugs, namely:

- the first drug includes the following components in the group consisting of: beta-blockers, sympatholytics with adrenolytic action, an H1 histamine receptor antagonist, antianginal drug with anti-ischemic effect, serotonergic drug, antipsychotic drug with hypothermic effect, antithyroid drug, a preparation containing magnesium ions and a pharmaceutically acceptable solvent or solvents.

- the second drug includes a fat emulsion of lipofundin, smoflipid, interlipid, saturated with an inert gas.

In this case, the first preparation contains the following ratio of components, wt. %:

propranolol beta blocker 0.261-0.317 wt. % sympatholytic reserpine 0.052-0.063 wt. % antianginal drug ivabradine 0.261-0.317 wt. % antihistamine diphenhydramine 0.261-0.317 wt. % hypothermic antipsychotic drug action of pericyazin 0.209-0.254 wt. % serotonergic serotonin hydrochloride 0.261-0.317 wt. % propithyl antithyroid 0.261-0.317 wt. % magnesium sulfate 1.569-1.904 wt. % pharmaceutically acceptable solvents 96.865-96.194 wt. %

As pharmaceutical acceptable solvents, physiological saline, Ringer's solution can be used.

In this case, the second drug includes a 20% fat emulsion of lipofundin, smoflipid, interlipid saturated with inert gas xenon.

1 ml of lipofundin emulsion contains from 0.54 ml to 2.2 ml of xenon. The ratio of the volumes of the solution of the first drug and the second in the tool is from 1/1 to 1/3.

Brief Description of the Drawings

FIG. 1. Growth dynamics of Ehrlich solid ascites carcinoma (AKE) in three groups of BALB / C mice. Where: 1. “tumor” (animals with AKE), 2. “Tumor + FITS + hypoxia” (animals with AKE, which were introduced into FITS on the 5th day after tumor formation, followed by 2-hour hypoxia) and 3. “Tumor + FITS + increasing hypoxia ”(animals with AKE, which were introduced into FITS on the 4th day after tumor formation followed by 2-hour hypoxia, the animals were introduced into a state of hypoxia slowly).

* - significant difference from the group "control" p≤0.05,

where: FITS is a pharmacologically induced torpor-like state.

The ordinate axis is the thickness of the paw (right) with the inoculated tumor in% of the start of the experiment. The abscissa is the duration of the experiment, days.

FIG. 2. The death of animal tumor carriers of three groups. Where: 1. "tumor" (animals with AKE); 2. “Tumor + FITS + hypoxia” (animals with AKE, which were introduced into FITS on the 5th day after the formation of the tumor, followed by 2-hour hypoxia); 3. “Tumor + FITS + increasing hypoxia” (animals with AKE, which were introduced into FITS on the 4th day after tumor formation, followed by 2-hour hypoxia, animals were introduced into a state of hypoxia slowly).

The y-axis is the number of surviving animals in% of the start of the experiment.

The abscissa is the duration of the experiment, weeks.

The implementation of the invention

In the process of developing a method for increasing the stability of a mammalian organism at the initial stage of tumor development, it was found that the solution to this problem is associated with the need to use means for a general slowdown in metabolic processes, accompanied by a drop in oxygen consumption, as well as creating an air environment low in relation to the host organism with a low oxygen level. In this case, additional hypoxic reactions of the body are formed that do not violate blood saturation, but increase the body's resistance to tumor development.

It was found that the use of an agent containing an active agent in the form of xenon gas immobilized in a carrier forms not only the possibility of slowing down metabolic processes, but also causes the body to resist external hypoxia. This leads to inhibition of tumor growth, with the subsequent introduction of the mammal into a state of additional hypoxia by reducing the level of oxygen in the environment to 5%.

The specified product contains a pharmaceutical mixture consisting of eight drugs: adrenergic blocker, sympatholytic, histamine H1 receptor antagonist, If-channel inhibitor of the sinus node, serotonergic drug, antipsychotic drug with hypothermic effect, antithyroid drug, preparation containing magnesium ions, and saturated fat emulsion xenon gas. The use of fat emulsion provides a prolonged release of xenon into the body of a mammal.

It is known that the adrenolytic agent propranolol is included in the group of adrenergic blockers, which also includes: cartolol, nadolol, penbutolol, pindolol, propranolol, sotalol, timolol, carvedilol, labetalol, metoprolol, atenolol, esmolol, acebutaloline, and others. selectively inhibiting If-channels of the sinus node, use ivabradine. Diphenhydramine is a member of the group of sedatives that block histamine H1 receptors and cholinergic receptors of autonomic nerve nodes. Propicyl is used as an antithyroid drug that suppresses the intrathyroid peroxidase system. Periciazine is used as a drug with a pronounced antiemetic, anticholinergic, hypothermic and sedative effect, moderately expressed adrenolytic and extrapyramidal effect. The main pharmacological property of reserpine is its sympatholytic effect, due to the effect of depletion of catecholamine depots with their subsequent destruction due to the inactivating effect of monoamine oxidase (MAO). Magnesium sulfate in the composition is used as a non-specific inhibitor of NMDA receptors to prevent excitotoxicity provoked by hyperactivation of these receptors during hypothermia.

A carrier in which xenon gas is immobilized is a fat emulsion selected from the group consisting of lipofundin, smoflipid, interlipid.

Use emulsion solutions in a concentration of 20%.

The ratio of the volumes of the solution of the first drug and the volume of the solution of the second drug is from 1/1 to 1/3. A ratio of 1/1 is preferred.

The concentration of inert xenon gas in a solution of a fat emulsion when it is saturated under a pressure of 2 atm is from 0.54 ml to 2.2 ml in 1 ml of a 20% fat emulsion.

Saline, Ringer's solution, and other balanced saline solutions are used as pharmaceutically acceptable solvents.

It is advisable to administer the agent directly to the mammal when it becomes necessary to save the body’s energy resources. In the proposed tool, instead of the existing and listed drugs, their analogues can be used both in the pharmacological group and in the effect exerted.

It was also found that the most preferred variant of the method for inhibiting tumor growth is that, along with the use of the agent, the mammal with the tumor is additionally introduced into the conditions of immersion in a state of hypoxia, in which the external conditions for the mammal are formed by lowering the oxygen level to 5 % for 35-40 minutes and keeping in these hypoxic conditions for 1.5-3 hours.

The formation of a pharmacologically induced torpor-like state (FITS) of the animal with the subsequent use of external hypoxic conditions can increase the antitumor potential of mice with a solid model of Ehrlich ascites carcinoma by slowing down tumor growth, stimulating pro-apoptotic mechanisms in tumor tissue cells, and also by activating antitumor immunity in mice .

The method of inhibiting tumor growth was tested in experiments on white clinically healthy male Balb / c mice weighing 20-25 grams, which were divided into groups of 10 animals each. The following examples include, but are not limited to, the use of analogues of active substances that are part of the tool. Before the experiment, control physiological parameters were recorded for 0.5 h. Rectal temperature (Tr) and heart rate (HR) were measured in mice, which averaged Tr = 37.5 ° C ± 0.3 ° C and heart rate 692 ± 22 beats / min. In FITS in mice, the average temperature was Tr = 23.3 ° C ± 0.2 ° C before being placed in a hypoxic chamber; average heart rate -214 ± 10 beats / min.

Example

The solid form of Ehrlich ascites carcinoma is formed by injection of 3 * 10 ⁶ cells into the hind paw of Balb / c mice. On the 3-4th day after this injection, a thickening of the paw of the animal was observed, and by the 20th day of the experiment, the paw with the tumor without treatment increased by about 3 times. On the 4th-5th day after the formation of the tumor, the tumor-bearing mice are given a single intraperitoneal injection of an agent containing a pharmaceutical composition consisting of eight preparations in combination with a carrier based on a fat emulsion and saturated with an inert gas.

The product includes a pharmaceutical composition containing a mixture of eight drugs in a volume of 0.5 ml in the following ratio of components: Propranolol - 0.289 wt. % (1.5 mg / kg), Ivabradine - 0.289 wt. % (1.5 mg / kg), diphenhydramine - 0.289 wt. % (1.5 mg / kg), Reserpine - 0.058 wt. % (0.3 mg / kg), Serotonin hydrochloride - 0.289 wt. % (1.5 mg / kg), Periciazine - 0.231 wt. % (1.2 mg / kg), Propitsil - 0.289 wt. % (1.5 mg / kg), magnesium sulfate - 1.737 wt. % (9 mg / kg). Prepare a 20% lipofundin emulsion. As pharmaceutical acceptable additives use physical. solution, Ringer's solution and other pharmaceutically acceptable solvents.

The first group of mice is used for control and not treated.

The second group of mice is exposed to an agent for the formation of a pharmacologically induced torpor-like state by a single intraperitoneal administration of an agent designed for a mouse weighing 25 grams, in which 0.015 ml of a solution of the composition and 0.015 ml of lipofundin fat emulsion saturated with xenon active agent. Mice are placed in a chamber in which hypoxic conditions are created with an oxygen level of 5%.

The third group of mice is exposed to an agent for the formation of a pharmacologically induced torpor-like state by intraperitoneal administration of an agent designed for a mouse weighing 25 grams, in which 0.015 ml of a solution of the composition and 0.015 ml of lipofundin fat emulsion saturated with xenon active agent. Mice are placed in a sealed chamber with normal air and begin to slowly and gradually displace oxygen from the air with nitrogen to 5% over a period of 35-40 minutes. After which the animals are kept in a hypoxic environment at an oxygen level of about 5% for 2 hours.

To evaluate the antitumor effect, a comparison of the indicators of three groups of mice is used according to the dynamics of changes in weight, body temperature, tumor growth, and also the life expectancy of tumor-bearing animals.

Figure 1, 2 shows the results of testing a method of treating a tumor with combined exposure to a pharmacological agent containing a composition of eight drugs and a lipofundin emulsion saturated with an inert gas, followed by exposure of the animal in hypoxic air.

In FIG. Figure 1 compares the growth dynamics of Ehrlich solid ascites carcinoma (AKE) in three groups of BALB / C mice. The first graph (marked with squares on the line) shows the dynamics for mammals with AKE without the use of therapeutic effects. The second graph (marked by rhombs on the line) shows the results of the growth dynamics of Ehrlich solid ascites carcinoma in mammals, which were administered a pharmacological agent on the 5th day after the formation of the tumor, followed by placement of the animals for 2 hours in a hypoxic environment with an oxygen content of 5%. The third graph (marked by triangles on the line) shows the growth dynamics of Ehrlich solid ascites carcinoma in mammals, which were administered a pharmacological agent on the 4th day after tumor formation, followed by placement of animals in a chamber with a slow decrease in oxygen in the medium (increasing hypoxia) to 5% per 35-40 minutes and keeping under these hypoxic conditions for two hours.

In FIG. 2 shows data on the survival of tumor-bearing animals of the three groups.

1. Black rectangles - “tumor” (animals with AKE).

2. Gray rectangles - “Tumor + FITS + hypoxia” (animals with AKE, which were introduced into FITS on the 5th day after the formation of the tumor, followed by 2-hour hypoxia).

3. White rectangles - “Tumor + FITS + increasing hypoxia” (animals with AKE, which were introduced into FITS on the 4th day after the formation of the tumor, followed by 2-hour hypoxia, the animals were introduced into a state of hypoxia slowly).

According to the experiment shown in FIG. 2 in the control group "tumor" in the tenth week of the experiment, 90% of the animals die from the tumor. Of the animals of the second group “tumor + FITS + hypoxia”, which received an injection of the agent and then placed in a medium containing 5% oxygen, about 20% of the animals survived at 10 weeks of the experiment. The number of animals of the third group “tumor + FITS + increasing hypoxia” surviving at 10 weeks is 55%.

The experimental data show that the use of the drug and the subsequent use of external hypoxia lead to a sharp inhibition of tumor growth, which allows to increase the time during which, if necessary, surgical treatment or chemotherapy can be used.

This example includes, but is not limited to, the scope of the invention.

The disclosed invention relates to a method of using an agent for the formation of pharmacologically induced hypometabolism and hypothermia in combination with hypoxia to inhibit the growth and development of tumors.

The use of an active agent, in the form of xenon gas, immobilized in the carrier of the agent, as a drug to increase the body's resistance to hypoxia, allows prolonging the stable life support of the body in an environment with low oxygen levels and allows for pronounced regression of the tumor at the first stage of its growth for subsequent execution radical surgery. At the same time, the possibility of reversible restoration of all functions of a mammal while restoring the level of oxygen in the environment remains. The proposed tool increases the life expectancy of mammals with malignant tumors and provides the speediest recovery of the body after treatment. The invention may find application in medicine to increase the stability of the body in the treatment of cancer.

Sources of information

1. ST. Kozlov et al. METHOD FOR TREATING ONCOLOGICAL LUNG CANCER PATIENTS, RF patent No. RU 2481091 (05/10/2013).

2. Yu.A. Barsukov et al. METHOD FOR TREATING DIRECT GUT CANCER, RF patent No. RU 2414936 (03/27/2011).

3. G.Yu. Gredasov METHOD FOR TREATING ONCOLOGICAL PATIENTS AND A DEVICE FOR ITS IMPLEMENTATION, RF patent No. RU 2326701 (06.20.2008).

4. J.A. Fox Method for inducing hypothermia for treating neurological disorders, US Pat. No. 6736837 (2004-05-18).

5. C.C. LEE, J. ZHANG METHODS AND COMPOSITIONS FOR INDUCING TORPOR IN A SUBJECT, US Application No. US 2007213295 (2007-09-13).

6. M.V. ROTH METHODS, COMPOSITIONS AND DEVICES FOR INDUCING STASIS IN CELLS, TISSUES, ORGANS, AND ORGANISMS, US Patent No. US 7993681 (2011-08-09).

Claims (11)

1. A method of inhibiting tumor growth in a mammal by forming hypometabolism and hypothermia, in which a means is used to form hypothermia and hypometabolism, which includes 20% lipofundin fat emulsion saturated with inert gas xenon, beta-blocker propranolol, sympatolytic reserpine, antianginal drug ivabradine , antihistamine, diphenhydramine, antipsychotic with periciazine, serotonergic serotonin hydrochloride, antithyroid medication your propicyl, magnesium sulfate and a pharmaceutically acceptable solvent from the group: physiological saline, Ringer's solution, while the product itself contains a combination of drugs, namely the first drug having the following ratio of components, wt. %: propranolol beta blocker  0.261-0.317 sympatholytic reserpine 0.052-0.063 antianginal drug ivabradine 0.261-0.317 antihistamine diphenhydramine 0.261-0.317 hypothermic antipsychotic drug action of pericyazin 0.209-0.254 serotonergic serotonin hydrochloride 0.261-0.317 propithyl antithyroid 0.261-0.317 magnesium sulfate 1,569-1,904 pharmaceutically acceptable solvents 96,865-96,194, and a second preparation, which contains 20% lipofundin fat emulsion saturated with inert gas xenon, where 1 ml of 20% lipofundin fat emulsion contains 0.54 ml to 2.2 ml xenon when saturated at a pressure of 2 atm, the agent injecting a mammal, and after the introduction of the drug, the mammal is subjected to additional hypoxia due to a decrease in oxygen in the environment to the level of 5% and incubated under these conditions for 1.5-3 hours. 2. The method according to p. 1, where additional mammalian hypoxia is formed by the method of displacing oxygen from the surrounding air for 35-40 minutes to a level of 5%. 3. The method according to p. 1, where the replacement of oxygen in the environment of the mammal is carried out by a gas of high purity nitrogen. 4. The method according to p. 1, where the ratio of the volumes of the solution of the first drug and the volume of the second drug is from 1/1 to 1/3. 5. Means for implementing the method according to PP. 1-4, which contains a combination of drugs, namely the first drug having the following ratio of components, wt. %: propranolol beta blocker 0.261-0.317 sympatholytic reserpine 0.052-0.063 antianginal drug ivabradine 0.261-0.317 antihistamine diphenhydramine 0.261-0.317 hypothermic antipsychotic drug action of pericyazin 0.209-0.254 serotonergic serotonin hydrochloride 0.261-0.317 propithyl antithyroid 0.261-0.317 magnesium sulfate 1,569-1,904 pharmaceutically acceptable solvents 96,865-96,194, and a second preparation that contains 20% lipofundin fat emulsion saturated with inert xenon gas. 6. The tool according to claim 5, where the ratio of the volumes of the solution of the first drug and the volume of the second drug is from 1/1 to 1/3. 7. The tool according to claim 5, where 1 ml of a 20% lipofundin fat emulsion contains from 0.54 ml to 2.2 ml of xenon when saturated at a pressure of 2 atm.

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