JP7361076B2 - testosterone booster - Google Patents

Description

The present invention relates to a testosterone-increasing agent that has the effect of increasing testosterone in the human body. The testosterone increasing agent of the present invention is characterized by containing nicotine as an active ingredient.

It is known that not only women but also men experience menopausal symptoms. In men, this menopausal disorder is also called Late-Onset Hypogonadism (LOH syndrome). "Menopause" refers to the period when sex hormones are lower than normal, which causes various problems not only physically but mentally. Psychological symptoms of andropause include depression, anxiety, loss of interest and motivation, inability to sleep, and irritability. On the other hand, physical symptoms include decreased sexual function, frequent urination, muscle pain, joint pain, sweating, hot flashes, and easy fatigue.
The big difference between menopausal symptoms in men and women is that women's menopausal symptoms often develop in the 10 years before and after menopause, whereas men's menopause is more affected by the environment, and the timing, duration, and extent of hormone decline. It is recognized that there are considerable individual differences in For men, the onset of menopausal symptoms is generally after the age of 40, but it may not appear for the first time until the age of 60 to 70.

Male hormone replacement therapy is widely used as a treatment for men. This treatment involves injecting male hormones about once a month, and it has been found that aging male symptom (AMS) score indicators often improve.
Another method of male hormone replacement therapy is to apply ointments. However, in Japan, only the injection method is currently approved by the Ministry of Health and Welfare. For this reason, it is difficult to say that male hormone replacement therapy is widely used around the world, as injections can be painful and hospital visits can be troublesome.
In some foreign countries other than Japan, treatments such as individuals applying testosterone cream or gel to the skin or administering testosterone to the nose may be popular, but in any case, in Japan, these treatments are is not covered by insurance. Therefore, a method that does not rely solely on direct injection or application of testosterone is widely desired.

By the way, treatments for prostate cancer often include treatments that remove testosterone from the human body. In such cases, it is known that the testosterone level in the brain also decreases, which may increase the probability of developing dementia.

Prior Art Document Patent Document 1 listed below describes a testosterone-increasing agent using tomato extract.
Further, Patent Document 2 listed below describes a testosterone increasing agent using NMDA (N-methyl-D-aspartic acid).
Further, Patent Document 3 listed below describes a testosterone increasing agent containing vitamin K.
Further, Patent Document 4 listed below describes a testosterone increasing agent using an onion extract.
Further, Patent Document 5 listed below describes the structure of a transdermal tobacco alkaloid patch for transdermally administering tobacco alkaloid components.
Various documents have described that so-called male menopausal symptoms can be improved by introducing testosterone into the body. For example, there are descriptions in the following non-patent documents 1, 2, 3, 4, and 5.

Furthermore, the following Non-Patent Documents 6 and 7 describe that transdermal administration such as a nicotine patch or suction using a low-temperature aerosol causes less burden on the patient.
Furthermore, Non-Patent Document 8 below reports that when a high concentration of nicotine of 500 μM acts on GT1-7 cells, GnRH secretion is suppressed.
Furthermore, Non-Patent Document 9 listed below reports that GnRH secretion is observed when a high concentration of nicotine of 300 μM or more is applied to GT1-7 cells, but no GnRH secretion can be observed at a lower concentration of 100 μM.

Furthermore, Non-Patent Document 10 below reports that nicotine acts on the testicles and causes damage to the steroid synthesis system, sex cell development, etc.
Furthermore, Non-Patent Document 11 listed below describes a specific increase in nicotine concentration in serum when a human smokes cigarettes.

Japanese Patent Application Publication No. 2017-105746 Japanese Patent Application Publication No. 2011-225474 Re-table No. 2007-148494 Japanese Patent Application Publication No. 2007-210918 Special Publication No. 2009-519246

K. L. Edinger, C. A. Frye, Psychoneuroendocrinology (2005) 30, 418?430Testosterone’s anti-anxiety and analgesic effects may be due in part to actions of its 5a-reduced metabolites in the hippocampus E. R. Rosario et al. JAMA, (2004) 292, 1431-1432 Age-Related Testosterone Depletion and the Development of Alzheimer Disease Y. Hatanaka et al., Brain Res. (2015) 1621, 121-132Rapid increase of spines by dihydrotestosterone and testosterone in hippocampal neurons: Dependence on synaptic androgen receptor and kinase networks Y. Hatanaka et al., Biochem. Biophys. Res. Comm. (2009) 381, 728?732Androgen rapidly increases dendritic thorns of CA3 neurons in male rat hippocampus Y. Hojo et al., Endocrinology (2009) 150, 5106-5112Comparison between Hippocampus-Synthesized and Circulation-Derived Sex Steroids in the Hippocampus H. Zuo, M. Ye, and M. Davies, Current Separations (1996) 15:2 Monitoring Transdermal Delivery of Nicotine Using In Vivo Microdialysis Sampling D. Yuki et al. Regulatory Toxicology and Pharmacology (2017) 87, 30-35 Pharmacokinetics of nicotine following the controlled use of a prototype novel tobacco vapor product Elio Messi et al. (2018) Molecular and Cellular Endocrinology 460, 209-218The alpha-7 nicotinic acetylcholine receptor is involved in a direct inhibitory effect of nicotine on GnRH release: In vitro studies Y. Arai et al. (2017) J. Physiol. Sci. 67:313?323Subunit profiling and functional characteristics of acetylcholine receptors in GT1-7 cells Jana et al. (2010) TOXICOLOGICAL SCIENCES 116, 647?659Nicotine Diminishes Testicular Gametogenesis, Steroidogenesis, and Steroidogenic Acute Regulatory Protein Expression in Adult Albino Rats: Possible Influence on Pituitary Gonadotropins and Alteration of Testicular Antioxidant Status Kalra et al. (2014) CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY,Vol.11, p. 563?568, Immunosuppressive and Anti-Inflammatory Effects of Nicotine Administered by Patch in an Animal Model GlaxoSmithKline Consumer Healthcare Japan K.K. (2019) Instructions for use of smoking cessation aid, transdermal nicotine preparation, Nicotinel (registered trademark) TTS, pharmacokinetics items Iwamoto et al. (2009) Int. Journal of Urology16, 168-174, Late-onset hypogonadism (LOH) and androgens: Validity of the measurement of free testosterone levels in the diagnostic criteria in Japan

As described above, the reality is that testosterone injection technology, which is an effective treatment for male menopausal symptoms, is not widely used in the world.
The present invention has been made in view of the above-mentioned problems, and an object of the present invention is to provide a simpler technique for injecting testosterone to realize effective treatment for menopausal disorders in men. It is thought that by realizing a simple method, it will be possible to further spread the use of testosterone injection technology throughout the world.

Nicotine is a type of plant base (alkaloid) and is mainly contained in tobacco leaves. Furthermore, nicotine is an agonist that acts on nAChR (nicotinic acetylcholine receptor), which is a nicotinic acetylcholine receptor present in, for example, the brain. Here, nAChR is one type of AChR, which is a receptor for acetylcholine, which is a major neurotransmitter in the central nervous system, and is an ion channel type receptor.

Nicotine is known to primarily bind to receptors (nAChR) in the brain and promote the release of neurotransmitters (dopamine, adrenaline, β-endorphin).
The inventor of this application focused on the effect of increasing testosterone by ingesting this nicotine into the body, and believed that if this nicotine could be easily introduced into the body, it would be possible to easily increase testosterone in the body. It is something. That is, the present invention provides a new use for nicotine.

At present, sufficient research has not been conducted on the increase in testosterone caused by ingesting nicotine and its associated effects on suppressing memory decline and antidepressant effects. Developing its uses will be a future challenge.

In order to achieve this objective, the testosterone increasing agent of the present invention is characterized by containing nicotine as an active ingredient. Nicotine acts on nAChR, a nicotinic acetylcholine receptor in the hypothalamus of the brain, releases progestin (LH), and increases testosterone synthesis in the testis. The present invention relates to a method using such a principle. Incidentally, nicotine itself is used for smoking cessation treatment by means of a nicotine patch, for example, and is a substance used in so-called good therapy.

About Testosterone Testosterone is a type of male hormone and is widely involved in maintaining cognitive function, anti-depression, building muscle, flexibility of blood vessels, sexual function, etc. The amount of testosterone secreted in the male body decreases with age. It has been pointed out that when blood testosterone levels decrease as a result, depressive symptoms appear, memory, concentration, and motivation decline, and muscle strength and male function decline. In recent years, the number of patients with age-related hypogonadism and andropause, in which the above-mentioned symptoms appear due to a decrease in the amount of testosterone in the blood, has been increasing.

According to the testosterone-increasing agent of the present invention, it is possible to increase the amount of testosterone secreted, thereby improving and alleviating various symptoms such as memory loss and depressive symptoms in the brain hippocampus caused by a decrease in testosterone concentration. Such effects of testosterone itself are described in the above-mentioned non-patent documents 1, 2, 3, and 4. Kawato, a researcher at the University of Tokyo, has shown that when the blood testosterone concentration increases, the testosterone concentration in the hippocampus, which is the brain's memory center, increases to about 1.3 times the blood testosterone concentration at that time. Laboratory research results show that. Therefore, it can be inferred that increasing blood testosterone levels is highly effective in improving memory and depressive symptoms.

Specifically, the present invention has the following configuration.
(1) In order to solve the above problems, the present invention is a testosterone increasing agent characterized by containing nicotine as an active ingredient.

(2) The present invention is also a testosterone increasing agent according to (1), characterized in that the nicotine is extracted and purified from tobacco leaves.

(3) The present invention is also a testosterone increasing agent according to (1), characterized in that the nicotine is chemically synthesized.

(4) The present invention also provides a testosterone increasing agent according to any one of (1) to (3), which has a patch structure for transdermal administration to the human body. It is a drug.

(5) The present invention also provides a testosterone increasing agent according to any one of (1) to (3), which is administered to the human body by inhalation using a low-temperature aerosol. be.

(6) Moreover, the present invention is a testosterone increasing agent according to any one of (1) to (3), which is characterized in that it is administered to the human body by nasal spray.

Various embodiments of the present invention are described in preferred embodiments of the present invention described below.
The testosterone increasing agent of this embodiment, which will be described later, contains nicotine as an active ingredient. The nicotine used in this embodiment may be extracted and purified from tobacco leaves by a conventional method, or may be a commercially available nicotine produced by a chemical synthesis method.

Furthermore, when using the testosterone increasing agent of the present embodiment, which will be described later, the administration method may be, for example, transdermal administration, inhalation, or the like. Preferably, transdermal administration such as a nicotine patch or inhalation using low-temperature aerosol may be used, since it places less burden on the patient. Alternatively, nasal sprays or the like may be used. These techniques are described in, for example, the above-mentioned non-patent documents 6 and 7. A nicotine patch that is widely known as a so-called smoking cessation aid may be used as is.

As explained above, according to the present invention, testosterone in the body can be effectively increased. Therefore, it is possible to provide a testosterone-increasing agent that contributes to the effective treatment of male menopausal symptoms.

It is a graph showing the change (increase) in serum testosterone concentration of a rat when a nicotine patch is applied to the back of the rat for 3 hours. It is a graph showing the change (increase) in nicotine concentration in the serum of a rat when a nicotine patch is applied to the back of the rat for 3 hours. It is a graph showing the change (increase) in the LH concentration of rat serum when a nicotine patch is applied to the back of the rat for 3 hours. It is a table showing changes in testosterone concentration in human saliva when a nicotine patch is applied to the human abdomen for 3 hours.

The inventors of the present invention verified the effects of nicotine administration using a nicotine patch using rats and humans.
First, when a nicotine patch is applied to the body surface of a rat or human, the nicotine concentration in the serum of the rat or human increases for the first 1 to 3 hours after application. Then, over the next 10 hours, the nicotine concentration in rat and human serum changes slowly, but the nicotine concentration is maintained at about 0.5 to 1.2 times the initial concentration (Non-patent Document 6). , 7, 12).

In this way, by applying a nicotine patch to the body surface, the nicotine concentration in the blood can be maintained at a high concentration for a certain period of time, and in proportion to that concentration, the excitatory nicotine dependence that exists in the hypothalamus of the brain It is believed that acetylcholine receptors (nAChRs) are activated. If nAChR exists in the hypothalamic GnRH (Gonadotropin releasing hormone) nerve, it is possible that the nAChR is activated and the released GnRH acts on the pituitary gland, releasing progestin (LH) into the blood. Highly sexual. It is then thought that LH enters the bloodstream and reaches the testes, increasing testosterone synthesis, and ultimately increasing the blood testosterone concentration.

According to these estimates, it is thought that the effect of increasing testosterone levels will be visible after about 3 hours have passed after applying the nicotine patch. Therefore, it is expected that nicotine will be effective in increasing the blood testosterone concentration by repeatedly acting over a short period of time, rather than by chronically acting on it for many days in a row.

Now, nAChR is expressed in various parts of the hypothalamus (arcuate nucleus, supraoptic nucleus, paraventricular nucleus, paraventricular nucleus, etc.), but it is difficult to accurately identify the nerves in which it is expressed. It is difficult to do so. There are various reasons for this, but basically it is because the hypothalamus contains many nerves and secretes hormones. For example, the GnRH nerve that secretes gonadotropin-releasing hormone GnRH, the GHRH nerve that secretes growth hormone-releasing hormone, the TRH nerve that secretes thyroid-stimulating hormone-releasing hormone, the CRH nerve that secretes corticotropin-releasing hormone, etc. There are many types of nerves, more than 10 types. Furthermore, upstream of the GnRH nerve, there is a kisspeptin nerve that secretes kisspeptin. Among these, it has not been clearly identified in which nerves nAChR is expressed.

There have been no rigorous reports regarding the expression of nAChR in hypothalamic GnRH neurons and its effects. The reason for this is that it is difficult to selectively stimulate only the GnRH nerves in the hypothalamus or to isolate only the GnRH nerves from the hypothalamus for genetic analysis.
The indirect method currently used is to create GT1-7 cells, which are cancerous cultured cells derived from the hypothalamus, and conduct experiments using these cells for analysis. However, it is well known that when nerve cells become cancerous, mutations occur, such as the ion channel receptors present in the original nerve cells often becoming defective. There is a report that when a high concentration of nicotine of 500 μM acts on GT1-7 cells, GnRH secretion is suppressed (see Non-Patent Document 8). There is also a report that GnRH secretion is observed when a high concentration of nicotine of 300 μM or more is applied to GT1-7 cells, but that GnRH secretion cannot be observed at a lower concentration of 100 μM (see Non-Patent Document 9). Here, 500 μM is an extremely high concentration of nicotine, approximately 1000 times the concentration found in serum from regular cigarette smoking.

At present, there are no known reports that nicotine directly increases LH release from the pituitary gland. There is also no known report that nicotine increases LH secretion from the hypothalamus + pituitary complex. Furthermore, there are no known reports that nicotine acts on the hypothalamus to increase blood testosterone. Furthermore, there are no reports that nicotine acts directly on the testicles to increase blood testosterone.
On the contrary, there are reports that nicotine acts on the testicles and causes damage to the steroid synthesis system, sex cell development, etc. (see Non-Patent Document 10).

If nicotine is used continuously for a long period of time, it can cause nicotine dependence (dopamine secretion by acting on nAChR of the dopamine nerve in the nucleus accumbens), hyperadrenaline (adrenaline is secreted from the adrenal medulla, resulting in a continued state of excitement), and testicular function. There is a possibility that various side effects that impede health, such as damage to the skin, may occur (see Non-Patent Document 10). Therefore, it is considered preferable to act for a short time and multiple times in order to avoid side effects.

For the reasons explained above, it is considered effective from the viewpoint of efficiency and reduction of side effects to administer a testosterone increasing agent containing nicotine as an active ingredient not continuously but multiple times at predetermined intervals. Moreover, in this embodiment, as an administration method of the testosterone increasing agent, an example of transdermal administration using a patch (nicotine patch) is shown. However, transdermal administration using a gel or the like may be used, or inhalation administration via a low-temperature aerosol may be used. Additionally, nicotine may be administered by other methods as long as it can be administered. For example, it may be applied to the subject as a nasal spray.

When a male mammal, including a human, ingests the testosterone increasing agent of the present embodiment not continuously but multiple times at predetermined time intervals, the amount of testosterone increases due to nicotine. Therefore, the testosterone increasing agent of the present embodiment can be expected to be effective as a therapeutic or preventive agent for testicular hypofunction and testosterone deficiency caused by aging or environmental factors. In addition, the testosterone increasing agent of the present invention can improve or treat various diseases caused by a decrease in testosterone, particularly cognitive function, concentration, motivation, muscle pain, flexibility of blood vessels, sexual function, and urinary function. can be improved or treated.

Note that although nicotine is used as an example in this embodiment, the effects described above are not limited to nicotine alone, and other synthetic agonists that actuate nAChRs may also be used. An example is carbachol.

Example 1 is a description of rats.
In this Example 1, first, 13 Wistar rats (male, 12 weeks old) purchased from Tokyo Experimental Animals Co., Ltd. were prepared as experimental animals. The rat is a model that can be extrapolated to humans living a normal life.

The rats were reared in a breeding room controlled at a room temperature in the range of 22 to 24°C, humidity in the range of about 50%, and a light/dark cycle set to 12 hours (light period: 8:00 to 20:00). ing. The animals were reared for about a week using an automatic water supply device and given free access to tap water and basic feed (powdered feed CRF-1, manufactured by Oriental Yeast Co., Ltd.).

For the nicotine patch, the weight of the rat was measured, and a 3.5 mg nicotine patch (Nicotinel (registered trademark) 10, transdermal nicotine preparation, GlaxoSmithKline Consumer Healthcare Japan) was applied to each 350 g of rat body weight. It worked by pasting it on the back of a rat. The dosage is 10 mg/kg body weight (see Non-Patent Document 11).

Figure 1 shows the results before applying the nicotine patch and 3 hours after applying the nicotine patch (so-called after transdermal action). At each timing, the blood of the rat was collected, and testosterone was extracted from the serum according to the procedure shown below.

<Method for measuring testosterone in serum>
Blood collected from rats was centrifuged (3,000 rpm, 20°C, 10 minutes), and the serum in the upper layer was collected. Testosterone in the serum was measured by measuring chemiluminescence of a ruthenium complex using electrochemiluminescence immunoassay (ECLIA), and the serum testosterone concentration (nM) was calculated from this result.

As shown in Figure 1, the serum testosterone concentration increased from 8.1 ± 2.1 nM [control] to 20.3 ± 3.6 nM [patch] after applying the nicotine patch for 3 hours ( p<0.05).

Here, [control] is the result of measurement using blood collected from the rat's tail vein several days before applying the nicotine patch. [Patch] is the result measured using blood collected from the heart of the same rat after applying the nicotine patch for 3 hours. From the results shown in FIG. 1, it is clear that serum testosterone can be increased by a testosterone-increasing agent containing nicotine.

<Measurement of nicotine in serum>
Nicotine in serum was measured by gas chromatograph mass spectrometry (GC-MS). The measurement results are shown in FIG. In the graph of Figure 2, [control] indicates the results measured using blood taken from the rat's tail vein several days before applying the nicotine patch, and [patch] indicates the results obtained after measuring the control. The results are shown using blood collected from the heart of the same individual after applying a nicotine patch for 3 hours. Serum nicotine concentration increased from approximately 0 (zero) nM [control] to 212±27 nM (34±4 ng/mL) [patch].

<Measurement of LH in serum>
LH (Luteinizing hormone) in serum was measured by ELISA (Enzyme-Linked Immuno Sorbent Assay). The results are shown in FIG. Also in the graph of FIG. 3, the meanings of "control" and "patch" are the same as in the graphs of FIGS. 1 and 2 described above. The concentration of LH in serum increased from about 2.1 ng/mL [control] to about 2.9 ng/mL [patch].

This Example 2 is a description for humans. In Example 2, five adult males were used as human subjects.

The nicotine patch was administered to adults by applying a single nicotine patch (Nicotinel® 10, transdermal nicotine preparation, GlaxoSmithKline Consumer Healthcare Japan) to the human abdomen. . The dose was 17.5 mg. This corresponds to the amount that produces a salivary nicotine concentration equivalent to that when a human smokes one to two cigarettes a day (see Non-Patent Documents 11 and 12).

Approximately 2 mL of the human saliva was poured into a plastic centrifuge tube before applying the nicotine patch (at 1:00 p.m.) and 3 hours after applying the nicotine patch (so-called after the transdermal effect). It was collected and immediately stored in a -80 degree freezer. Testosterone was extracted from the saliva using the procedure shown below.

<Method for measuring testosterone in saliva>
After thawing saliva collected from humans, it was centrifuged (3,000 rpm, 4° C., 5 minutes), and the upper layer was collected. Testosterone in the upper fraction was measured using tandem mass spectrometry (LC-MS-MS), and the salivary testosterone concentration was calculated from the results.

Testosterone concentration in saliva was compared before applying the nicotine patch [control] and after applying the nicotine patch for 3 hours [patch]. Here, patch experiments on the same individual were conducted at intervals of several days or more. On different days, people's [control] testosterone levels varied slightly, and even within the same individual there were two cases in which the [control] levels of testosterone were lower and higher than average. Therefore, the cases of [control low value] and [control high value] were compared separately and shown in Table 1 (FIG. 4). In the case of [control low value], the concentration increased by 16.6%±3.1%. In the case of [control high value], the concentration decreased by 24% ± 7.7%. Here, the probability of [control low value] occurring is 15/20 times, which is overwhelmingly higher than the probability of [control high value] occurring 5/20 times, and the probability that the testosterone concentration will increase with the nicotine patch decreases. That's three times the probability.

Testosterone control in the same individual was divided into two cases: low and high levels, but low levels appear much more frequently than high levels. [Control] High values occur about 1/4 of the time in young people around 20 years old, but in elderly people around 70 years old, [Control] high values occur only about 1/11 times, which is almost always low. was found from a number of [control] salivary testosterone measurements. Considering these points, in elderly people with low testosterone levels (see Non-Patent Document 13), the nicotine patch effectively works as a testosterone-increasing agent.

As explained above, according to the present embodiment, it has been shown that nicotine can be used to effectively increase testosterone in the bodies of rats and humans. Therefore, a testosterone increasing agent containing nicotine as an active ingredient can effectively treat menopausal disorders in men.

Furthermore, the testosterone increasing agent according to the present embodiment can efficiently increase nicotine concentration by repeating short-term effects, and is therapeutically effective. That is, the testosterone increasing agent according to the present embodiment may be used for a short period of time (for example, 3 hours), and this may be repeated once or multiple times as appropriate. The cycle of this repetition may be long or short. It may also be applied "as needed" depending on the patient's symptoms.

Furthermore, the embodiments described above are examples of means for realizing the present invention, and should be modified or changed as appropriate depending on the object to which the present invention is applied, the device configuration, and various conditions. It should not be limited to the aspects of the embodiments.

Claims (3)

A testosterone increasing agent containing nicotine as an active ingredient , characterized in that it has a patch structure for transdermal administration to the human body. A testosterone increasing agent containing nicotine as an active ingredient , which is characterized in that it is administered to the human body by inhalation using a low-temperature aerosol. A testosterone increasing agent containing nicotine as an active ingredient , characterized in that it is administered to the human body by nasal spray.